WO2020123881A1

WO2020123881A1 - Boron containing compounds and their uses

Info

Publication number: WO2020123881A1
Application number: PCT/US2019/066100
Authority: WO
Inventors: Chun Yu Yu LIU; Yasheen Zhou; Stephen Yang; Cbunliang LIU; Yong-Kang Zhang
Original assignee: Boragen, Inc.
Priority date: 2018-12-12
Filing date: 2019-12-12
Publication date: 2020-06-18

Abstract

The present disclosure relates to boron-containing compounds, novel combiantions, and novel uses. The disclosure provides novel benzoxaborole compounds, pharmaceutical compositions, and associated methods of use. In particular, modeling suggests the benzoxaborole compounds disclosed herein are useful for inhibiting protein prenylation or ergosterol synthesis in a pathogenic microorganism. The benzoxaborole compounds according to the disclosure are further useful for the treatment of infectious microorganisms including pathogenic fungi and protozoan parasites, such as those that cause malaria.

Description

BORON CONTAINING COMPOUNDS AND THEIR USES

Cross-Reference to Related Applications

[001] The present application claims the benefit of United States Provisional

Application No. 62/778,871 , filed December 12, 2018, which is hereby incorporated by reference in its entirety.

Technical Field

[002] The present disclosure relates to boron-containing compounds, novel combinations thereof, and their novel uses. The disclosure provides compounds, pharmaceutical compositions, and associated methods of use. In particular, modeling suggests the benzoxaborole compounds disclosed herein are useful for inhibiting protein prenylation or ergosterol synthesis in a pathogenic microorganism. The benzoxaborole compounds according to the disclosure are further useful for the treatment of infectious microorganisms including pathogenic fungi and protozoan parasites, such as those that cause malaria.

Background

[003] Pathogenic infections stemming from parasites and fungi represent a major clinical problem worldwide. As an example, malarial disease, which is caused by protozoan parasites, in Africa has suffered increased drug resistance, increasing mortality rates, and lack of effective treatments. As another example, fungal diseases including Candida, Aspergillus, Cryptococcus, and Pneumocystis affect up to two million people worldwide each year. These infectious fungal diseases are associated with high morbidity and mortality, and the incidence rate of these infections is increasing

(Schmiedel, Yvonne and Zimmerli, Stefan., Swiss Med Wkiy. 146:w14281 , (2016)). In addition, anti-fungal therapies currently available suffer from lack of efficacy, drug interactions, and toxicity.

[004] Pathogenic fungi have an enormous impact on human health. Most people are aware of some of the superficial infections caused by fungi. These include skin and nail infections such as athlete’s foot and ringworm, predominantly caused by dermatophytes ( Trichophyton , Microsporum and Epidermophyton species). In fact, these superficial mycoses represent one of the most prevalent forms of human infection, as they affect 20%-25% of the world's population (Havlickova. Czaika, & Friedrich, 2008). Mucosal infections are also extremely common; tor example, vulvovaginal candidiasis (thrush) affects 75% of women of childbearing age and 5%-8% suffer from recurrent infections (Sobel, 2007). More seriously, certain fungal species can cause invasive diseases in humans. Collectively, these infections have been estimated to kill 1.5 million people per year (Brown et al., 2012). These life-threatening infections are caused primarily by species of the Candida, Aspergillus, Cryptococcus and Pneumocystis genera, which cause life-threatening infections in individuals with impaired immunity or other underlying conditions.

[005] These infections can be broadly divided into environmentally (e.g. Aspergillus and Cryptococcus ) or endogenously (e.g. Candida ) acquired. For example, Aspergillus fumigatus grows as a saprophyte in environmental niches such as compost and soil and subjects become intected via inhalation of spores. In contrast, Candida albicans is a member of the human microbiota (or, in this case, mycobiota) and invasive infections typically originate from colonising cells of the subject's own gastrointestinal tract.

Therefore, these different human fungal pathogens have evolved pathogenic potential independently, either in environmental niches or as colonisers of mucosal surfaces.

[006] Thus, there is an unmet need for novel safe and effective therapies for pathogenic infections that include both protozoan parasites and fungi.

Brief Summary

[007] As described in detail herein, the present disclosure includes benzoxaborole compounds that are effective inhibitors of pathogenic microorganisms including, inter alia, fungi and protozoan parasites.

[008] One embodiment of the present disclosure includes a method for treating a subject having a pathogenic infection stemming from a pathogenic microorganism comprising administering a therapeutically effective amount of a compound of formula (I):

wherein:

X¹ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C1 -C3 hydrocarbyl, C1 -C3 hydrocarbyl having 1 -7 halogen substitutions, and cyclopropyl; X^1’ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C1-C3 hydrocarbyl, C1-C3 hydrocarbyl having 1 -7 halogen substitutions, and cyclopropyl;

Z¹ is oxygen or sulfur;

Y¹ is selected from the group consisting of: oxygen, sulfur, and NH;

R¹ is selected from the group consisting of: hydrogen, unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C3-C6 cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^{1 a} substitutions, unsubstituted benzyl, benzyl having 1 -7 R^1a substitutions,

unsubstituted heteroaryl, heteroaryl substituted with one or more R^1a substitutions, unsubstituted heterocyclyl, and heterocyclyl substituted with one or more R^1a

substitutions;

each R^1a independently is selected from the group consisting of C₁-C₈ hydrocarbyl, NH₂, OH, NO2, CN, C1-C2 haloalkyl, OR⁶, SR⁶, SOR⁶, SO2R⁶, NHC(O)OR⁶, NR⁶R⁷, C(O)OR⁶, unsubstituted aryl, C3-C6 cyclohydrocarbyl, fluorine, chlorine, bromine, iodine, heteroaryl, and heterocyclyl;

R⁶ and R⁷ are independently selected from the group consisting of C1-C6 hydrocarbyl and substituted hydrocarbyl, or

R⁶ and R⁷ can be taken together with the nitrogen to which they are attached to form a ring of no more than 8 members; and

each of R⁸ and R⁹ independently is selected from the group consisting of: hydrogen, methyl, ethyl; or

R⁸ and R⁹ can be taken together to form a 3 - 6 membered ring,

or a salt, stereoisomer, enantiomer, or tautomer thereof.

[009] One embodiment of the present disclosure includes wherein the pathogenic infection is selected from a mucosal infection, systemic infection, invasive infection, and a superficial infection. One embodiment of the present disclosure includes wherein the pathogenic infection is selected from a fungal infection and a protozoan infection.

[0010] One embodiment of the present disclosure includes wherein the fungal infection is selected from Aspergillosis, Blastomycosis, Candidiasis, Chromomycosis,

Coccidioidomycosis, Cryptococcosis, Dermatophytoses, Histoplasmosis, Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis, Paracoccidioidomycosis, Penicillium marneffei, Phaeohyphomycosis, Pneumocystis pneumonia, Rhinosporidiosis,

Sporotrichosis, Trichosporonosis, and Zygomycosis. [001 1 ] One embodiment of the present disclosure includes wherein the compound is administered in an amount to perturb a prenylation pathway of the pathogenic microorganism. One embodiment of the present disclosure includes wherein the compound is administered in an amount to perturb the ergosterol biosynthetic pathway of the microorganism. One embodiment of the present disclosure includes a wherein the perturbation of the prenylation pathway results in an effect comprising reduction of protein prenylation in the in the pathogenic microorganism. One embodiment of the present disclosure includes wherein the perturbation of the ergosterol biosynthetic pathway results in an effect comprising reduction of ergosterol synthesis in the pathogenic microorganism.

[0012] One embodiment of the present disclosure includes wherein the compound is administered with a second antifungal agent to treat the fungal infection. One embodiment of the present disclosure includes wherein the second antifungal agent is an inhibitor of lanosterol demethylase. One embodiment of the present disclosure includes wherein the inhibitor of lanosterol demethylase is an imidazole or triazole antifungal agent. One embodiment of the present disclosure includes wherein the lanosterol demethylase inhibitor is selected from fluconazole, fosfluconazole, ketoconazole, oxiconazole, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, eberconazole, econazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, efinaconazole, fluconazole, fosfluconazole, terconazole, hexaconazole, isavuconazole, itraconazole, posaconazole, voriconazole, albaconazole, and ravuconazole. One embodiment of the present disclosure includes wherein the second antifungal agent is an inhibitor of leucyl t-RNA synthetase. One embodiment of the present disclosure includes wherein the inhibitor of leucyl t-RNA synthetase is is tavaborole or 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole. One embodiment of the present disclosure includes wherein the compound and the second antifungal agent are synergistic in inhibiting fungal growth.

[0013] One embodiment of the present disclosure includes wherein the protozoan infection is from a phylum selected from Sarcomastigophora, Apicomplexa, Microspora, and Ciliophora.

[0014] One embodiment of the present disclosure includes wherein the protozoan infection is selected from Plasmodium, Trypanosoma, Leishmania, Toxoplasma,

Eimeria, Neospora, Cyclospora, Giardia, Entamoeba, Dientamoeba, Naegleria, Acanthamoeba, Babesia, Isospora, Sarcocystis, Enterocytozoon, Balantidium,

Pneumocystis, and Cryptosporidium. One embodiment of the present disclosure includes wherein the protozoan infection causes malaria. One embodiment of the present disclosure includes wherein the malaria is caused by an infection with one or more protozoan organisms selected from P. falciparum, P. vivax, P. ovale, and P. malariae. One embodiment of the present disclosure includes wherein the compound is administered with a second anti-protozoan agent to treat the protozoan infection. One embodiment of the present disclosure includes wherein the second anti-protozoan agent is selected from metronidazole, nifurtimox, atovaquone, pentamidine, benznidazole, quinacrine, dehydroemetine, sodium stibogluconate, eflornithine, suramin, emetine, tinidazole, fenbendazole, iodoquinol, melarsoprol, meglumine antimonite, nitazoxanide, chloroquine, hydroxychloroquine, primaquine, mefloquine, quinine, pyrimethamine, and doxycycline.

[0015] One embodiment of the present disclosure includes a method of perturbing protein prenylation or ergosterol synthesis in a pathogenic microorganism comprising contacting the pathogenic microorganism with a compound of formula (I):

wherein:

X¹ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C1-C3 hydrocarbyl, C1-C3 hydrocarbyl having 1 -7 halogen substitutions, and cyclopropyl;

X^1’ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C1-C3 hydrocarbyl, C1-C3 hydrocarbyl having 1 -7 halogen substitutions, and cyclopropyl;

Z¹ is oxygen or sulfur;

Y¹ is selected from the group consisting of: oxygen, sulfur, and NH;

substitutions;

R⁸ and R⁹ can be taken together to form a 3 - 6 membered ring,

or a salt, stereoisomer, enantiomer, or tautomer thereof.

[0016] One embodiment of the present disclosure includes wherein the pathogenic microorganism is a fungi or a protozoan. One embodiment of the present disclosure includes wherein the pathogenic microorganism is contacted with an amount of the compound to perturb a prenylation pathway of the pathogenic microorganism. One embodiment of the present disclosure includes wherein the pathogenic microorganism is contacted with an amount of the compound to perturb a ergosterol biosynthetic pathway of the pathogenic microorganism. One embodiment of the present disclosure includes wherein the compound preturbs a prenylation enzyme comprising farnesyl diphosphate synthase (Erg20), geranylgeranyl transferase (Cdc 43), or farnesyltransferase or combinations thereof. One embodiment of the present disclosure includes wherein the compound preturbs farnesyl diphosphate synthase (Erg20). One embodiment of the present disclosure includes a wherein the compound preturbs geranylgeranyl transferase (Cdc 43). One embodiment of the present disclosure includes wherein the compound preturbs farnesyltransferase. One embodiment of the present disclosure includes wherein a prenylation enzyme has one or more mutations. One embodiment of the present disclosure includes wherein the pathogenic microorganism is contacted with a concentration of the compound of about 0.05 ppm to about 50 ppm. [0017] One embodiment of the present disclosure includes wherein the pathogenic microorganism is a fungi and is further contacted with a second antifungal agent. One embodiment of the present disclosure includes wherein the compound and second antifungal agent is provided in a molar ratio of about 1 :20 to about 20:1. One embodiment of the present disclosure includes wherein the second antifungal agent is a lanosterol demethylase inhibitor selected from fluconazole, fosfluconazole,

ketoconazole, oxiconazole, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, eberconazole, econazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, efinaconazole, fluconazole, fosfluconazole, terconazole, hexaconazole, isavuconazole, itraconazole, posaconazole, voriconazole, albaconazole, and ravuconazole. One embodiment of the present disclosure includes wherein the second antifungal agent is an inhibitor of leucyl t-RNA synthetase. One embodiment of the present disclosure includes wherein the inhibitor of leucyl t-RNA synthetase is is tavaborole or 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole.

[0018] One embodiment of the present disclosure includes wherein the pathogenic infection is selected from a fungal infection. One embodiment of the present disclosure includes wherein Y¹ is O or S. One embodiment of the present disclosure includes wherein Z¹ is S. One embodiment of the present disclosure includes wherein the compound is a compound of formula (II):

[0019] One embodiment of the present disclosure includes wherein X¹ is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, unsubstituted C1-C3 hydrocarbyl, and C1-C3 hydrocarbyl having 1-7 halogen substitutions. One embodiment of the present disclosure includes wherein X^1’ is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, unsubstituted C1-C3 hydrocarbyl, and C1-C3 hydrocarbyl having 1-7 halogen substitutions. One embodiment of the present disclosure includes wherein: when X¹ is hydrogen, X^1’ is other than hydrogen; and when X^1’ is hydrogen, X¹ is other than hydrogen. One embodiment of the present disclosure includes wherein R¹ is selected from the group consisting of: hydrogen, unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C3-C6 cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^1a substitutions, unsubstituted benzyl, benzyl having 1-7 R^1a substitutions, unsubstituted heterocyclyl, and heterocyclyl substituted with one or more R^{1 a} substitutions; and each R^1a independently is selected from the group consisting of C₁-C₈ hydrocarbyl, NH₂, OH, OR⁶, SR⁶, NHC(O)OR⁶, NR⁶R⁷, C(O)OR⁶, unsubstituted aryl, C3-C6 cyclohydrocarbyl, fluorine, chlorine, bromine, and iodine.

[0020] One embodiment of the present disclosure includes wherein the compound is selected from a compound in Table A:

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

or a salt, stereoisomer, enantiomer, or tautomer thereof.

[0021 ] One embodiment of the present disclosure includes wherein the pathogenic infection is a protozoan infection. One embodiment of the present disclosure includes wherein the compound is a compound of Formula (III):

[0022] One embodiment of the present disclosure includes wherein X¹ is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, unsubstituted C1 -C3 hydrocarbyl, and C1 -C3 hydrocarbyl having 1-7 halogen substitutions; and R¹ is selected from the group consistng of unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^1a substitutions, unsubstituted benzyl, and benzyl having 1 -7 R^1a substitutions.

[0023] One embodiment of the present disclosure includes wherein the compound is selected from Table B:

Table B

or a salt, stereoisomer, enantiomer, or tautomer thereof.

[0024] One embodiment of the present disclosure includes wherein the protozoan infection is selected from Plasmodium, Trypanosoma, Leishmania, Toxoplasma, Eimeria, Neospora, Cyclospora, Giardia, Entamoeba, Dientamoeba, Naegleria, Acanthamoeba, Babesia, Isospora, Sarcocystis, Enterocytozoon, Balantidium, Pneumocystis, and Cryptosporidium. [0025] One embodiment of the present disclosure includes wherein the protozoan infection causes malaria and the malaria is caused by an infection with one or more protozoan organisms selected from P. falciparum, P. vivax, P. ovale, and P. malariae.

[0026] One embodiment of the present disclosure includes wherein the pathogenic microorganism is contacted with a concentration of the combination of about 0.05 ppm to about 50 ppm.

[0027] One embodiment of the present disclosure includes a combination comprising a) a compound of formula (I):

wherein:

X¹ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C₁-C₃ hydrocarbyl, C₁-C₃ hydrocarbyl having 1 -7 halogen substitutions, and cyclopropyl;

X^1’ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C₁-C₃ hydrocarbyl, C₁-C₃ hydrocarbyl having 1 -7 halogen substitutions, and cyclopropyl;

Z¹ is oxygen or sulfur;

Y¹ is selected from the group consisting of: oxygen, sulfur, and NH;

R¹ is selected from the group consisting of: hydrogen, unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C₃-C₆ cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^{1 a} substitutions, unsubstituted benzyl, benzyl having 1 -7 R^1a substitutions,

unsubstituted heteroaryl, heteroaryl substituted with one or more R^1a substitutions, unsubstituted heterocyclyl, and heterocyclyl substituted with one or more R^1a substitutions;

each R^1a independently is selected from the group consisting of C₁-C₈ hydrocarbyl, NH₂, OH, NO₂, CN, C₁-C₂ haloalkyl, OR⁶, SR⁶, SOR⁶, SO₂R⁶, NHC(O)OR⁶, NR⁶R⁷, C(O)OR⁶, unsubstituted aryl, C3-C6 cyclohydrocarbyl, fluorine, chlorine, bromine, iodine, heteroaryl, and heterocyclyl;

R⁸ and R⁹ can be taken together to form a 3 - 6 membered ring,

or a salt, stereoisomer, enantiomer, or tautomer thereof; and

b) one or more additional agent selected from:

i) a demethylation inhibitor; and

ii) a leucyl-tRMA synthetase inhibitor.

[0028] One embodiment of the present disclosure includes wherein the compound of formua (I) is a compound of formula (IV):

[0029] One embodiment of the present disclosure includes wherein X¹ is halogen. One embodiment of the present disclosure includes wherein Y¹ is S. One embodiment of the present disclosure includes wherein Y¹ is O. One embodiment of the present disclosure includes wherein R¹ is selected from the group consisting of: unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C3-C6 cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^1a substitutions, unsubstituted benzyl, and benzyl having 1 -7 R^1a substitutions. One embodiment of the present disclosure includes wherein R¹ is selected from the group consisting of: unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1-17 R^1a substitutions, unsubstituted C₆ cyclohydrocarbyl, C₆ cyclohydrocarbyl having 1 -11 R^1a substitutions, unsubstituted phenyl, phenyl having 1 -5 R^{1 a} substitutions, unsubstituted benzyl, and benzyl having 1 -7 R^1a substitutions.

[0030] One embodiment of the present disclosure includes a wherein the compound is

, or a salt, stereoisomer, enantiomer, or tautomer thereof.

[0031 ] One embodiment of the present disclosure includes wherein the compound is

salt, stereoisomer, enantiomer, or tautomer thereof.

[0032] One embodiment of the present disclosure includes wherein the compound and additional agent is provided in a molar ratio of about 1 :20 to about 20:1 .

[0033] One embodiment of the present disclosure includes wherein the demethylation inhibitor is one or more of tebuconazole, cyproconazole, propiconazole and fluconazole.

[0034] One embodiment of the present disclosure includes wherein the leucyl-tRNA synthetase inhibitor is is 5-chioro-1-hydroxy-3H-2.1 -benzoxaboroie.

[0035] One embodiment of the present disclosure comprises:

stereoisomer, enantiomer, or tautomer thereof; and

b) tebuconazole.

[0036] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) cyproconazole.

[0037] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) propiconazole.

[0038] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) fluconazole.

[0039] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) 5-Chloro-1 -hydroxy-3H-2,1 -benzoxaborole.

[0040] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof;

b) tebuconazole.

[0041 ] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) cyproconazole. [0042] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) propiconazole.

[0043] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) fluconazole.

[0044] One embodiment of the present disclosure comprises:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) 5-chioro-1 -hydroxy-3H-2,1 -benzoxaboroie.

[0045] One embodiment of the present disclosure includes a method for treating a subject having a pathogenic infection stemming from a pathogenic microorganism comprising administering a therapeutically effective amount of a combination of the present disclosure.

[0046] In one embodiment, the pathogenic infection is selected from a mucosal infection, systemic infection, invasive infection, and a superficial infection. In one embodiment, the pathogenic infection is a fungal infection. In one embodiment, the fungal infection is selected from Aspergillosis, Blastomycosis, Candidiasis,

Chromomycosis, Coccidioidomycosis, Cryptococcosis, Dermatophytoses,

Histoplasmosis, Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis, Paracoccidioidomycosis, Penicillium marneffei, Phaeohyphomycosis, Pneumocystis pneumonia, Rhinosporidiosis, Sporotrichosis, Trichosporonosis, and Zygomycosis. In one embodiment, the subject is an animal. In one embodiment, the subject is a mammal. In one embodiment, the mammal is a human. [0047] One additional embodiment of the present disclosure includes each novel compound herein disclosed as composition of matter.

[0048] The capability to infect an immunocompetent human has arisen independently multiple times among three major fungal phyla: the Entomophthoromycota, the

Ascomycota, and the Basidiomycota. Reference is made to Kohler et al., The Spectrum of Fungi That Infects Humans, Cold Spring Harb Perspect Med. 2015 Jan; 5(1 ):

a019273, herein incorporated by reference with regard to human fungal infections. Moreover, a sufficiently immunocompromised host can be infected by hundreds of environmental fungal species that grow at human core temperatures. A predictable set of actors, however, is known to cause the most common invasive infections in immunocompromised individuals. Likewise, reference is made to Kohler et al., The Spectrum of Fungi That Infects Humans, Cold Spring Harb Perspect Med. 2015 Jan; 5(1): a019273, herein incorporated by reference with regard to human fungal infections.

[0049] In another embodiment, the compound of the present disclosure is administered in an amount to perturb a prenylation pathway of the pathogenic microorganism. In another embodiment, the compound of the present disclosure is administered in an amount to perturb the ergosterol biosynthetic pathway of the microorganism. In another embodiment, the perturbation of the prenylation pathway results in an effect comprising reduction of protein prenylation in the pathogenic microorganism. In another embodiment, the perturbation of the ergosterol biosynthetic pathway results in an effect comprising reduction of ergosterol synthesis in the pathogenic microorganism.

[0050] In one embodiment, the pathogenic infection is selected from a mucosal infection, systemic infection, invasive infection, and a superficial infection. In another embodiment, the pathogenic infection is selected from a fungal infection and a protozoan infection. In another embodiment, the fungal infection is selected from Aspergillosis, Blastomycosis, Candidiasis, Chromomycosis, Coccidioidomycosis, Cryptococcosis, Dermatophytoses, Histoplasmosis, Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis, Paracoccidioidomycosis, Penicillium marneffei, Phaeohyphomycosis, Pneumocystis pneumonia, Rhinosporidiosis, Sporotrichosis, Trichosporonosis, and Zygomycosis.

[0051 ] In another embodiment, the compound of the present disclosure is administered with a second antifungal agent to treat the fungal infection.

[0052] In another embodiment, the second antifungal agent is an inhibitor of lanosterol demethylase. In another embodiment, the inhibitor of lanosterol demethylase is an imidazole or triazole antifungal agent. In another embodiment, the lanosterol demethylase inhibitor is selected from fluconazole, fosfluconazole, ketoconazole, oxiconazole, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, eberconazole, econazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, efinaconazole, fluconazole, fosfluconazole, terconazole, hexaconazole, isavuconazole, itraconazole, posaconazole, voriconazole, albaconazole, and ravuconazole. In another embodiment, the second antifungal agent is an inhibitor of leucyl t-RNA synthetase. In another embodiment, the inhibitor of leucyl t-RNA synthetase is is tavaborole or 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole.

[0053] In another embodiment, the compound of the present dislcosure and the second antifungal agent are synergistic in inhibiting fungal growth.

[0054] In another embodiment, the protozoan infection is from a phylum selected from Sarcomastigophora, Apicomplexa, Microspora, and Ciliophora. In another

embodiment, the protozoan infection is selected from Plasmodium, Trypanosoma, Leishmania, Toxoplasma, Eimeria, Neospora, Cyclospora, Giardia, Entamoeba, Dientamoeba, Naegleria, Acanthamoeba, Babesia, Isospora, Sarcocystis,

Enterocytozoon, Balantidium, Pneumocystis, and Cryptosporidium.

[0055] In another embodiment, the protozoan infection causes malaria. In another embodiment, the malaria is caused by an infection with one or more protozoan organisms selected from P. falciparum, P. vivax, P. ovale, and P. malariae.

[0056] In another embodiment, the the compound of the present disclosure is administered with a second anti-protozoan agent to treat the protozoan infection. In another embodiment, the second anti-protozoan agent is selected from metronidazole, nifurtimox, atovaquone, pentamidine, benznidazole, quinacrine, dehydroemetine, sodium stibogluconate, eflornithine, suramin, emetine, tinidazole, fenbendazole, iodoquinol, melarsoprol, meglumine antimonite, nitazoxanide, chloroquine, hydroxychloroquine, primaquine, mefloquine, quinine, pyrimethamine, and doxycycline.

[0057] In another aspect, the disclosure includes a method for controlling, reducing, preventing, ameliorating, or inhibiting an infestation by a pathogen by applying an effective amount of a compound of the present disclosure, wherein the pathogen is selected from a group consisting of one or more fungi and one or more protozoa.

[0058] In another aspect, the disclosure includes a method for controlling, reducing, preventing, ameliorating, or inhibiting an infestation by a pathogen by applying a compound of the present disclosure, wherein the pathogen is selected from a group consisting of one or more fungi and one or more protozoa.

[0059] In yet another aspect, the disclosure includes a method for controlling or preventing an infestation of the pathogen by treating an animal with an effective amount of a compound of the present disclosure.

[0060] In yet another aspect, the disclosure includes a method for controlling or preventing an infestation of the pathogen by treating an animal with a compound of the present disclosure.

[0061 ] The preceding is a simplified summary to provide an understanding of some embodiments of the present disclosure. This summary is neither an extensive nor exhaustive over-view of the present disclosure and its various embodiments. The summary presents selected concepts of the embodiments of the present disclosure in a simplified form as an introduction to the more detailed description presented below. As will be appreciated, other embodiments of the present disclosure are possible utilizing, alone or in combination, one or more of the embodiment, aspects, or features set forth above or described in detail below.

Brief Description of the Drawings

[0062] The foregoing and other aspects of the embodiments disclosed herein are best understood from the following detailed description when read in connection with the accompanying drawings. For the purposes of illustrating the embodiments disclosed herein, there is shown in the drawings embodiments that are presently preferred, it being understood, however, that the embodiments disclosed herein are not limited to the specific instrumentalities disclosed. Included in the drawings are the following figures:

[0063] FIG.1 illustrates a, in accordance with an embodiment of the present disclosure; a schematic representation of a synthetic scheme for making the boron containing compounds of the present disclosure.

[0064] FIGS. 2A, 2B, and 2C represent tables of examples of biological activity of the boron containing compounds of the present disclosure, as individual embodiments of the present disclosure.

[0065] FIG. 3 represents a molecular docking model of the compound 44 in the binding pocket of the enzyme GGTasel

[0066] FIG. 4 represents a molecular docking model of the compound 45 in the binding pocket of the enzyme Erg 20. [0067] FIG. 5 is a table showing MIC results of compound 44 and 53 against various WT and resistance mutant strains of Saccharomyces cerevisiae. The MIC is indicated in ppm.

[0068] FIG. 6 is a set of synergy maps illustrating the results of synergy testing with one or more compounds of the present disclosure and commercially available fungicides.

[0069] FIG. 7 is a set of synergy maps illustrating the results of synergy testing with one or more compounds of the present disclosure and commercially available fungicides.

[0070] While embodiments of the present disclosure are described herein by way of example using several illustrative drawings, those skilled in the art will recognize the present disclosure is not limited to the embodiments or drawings described. It should be understood the drawings and the detailed description thereto are not intended to limit the present disclosure to the form disclosed, but to the contrary, the present disclosure is to cover all modification, equivalents and alternatives falling within the spirit and scope of embodiments of the present disclosure as recited by the appended claims.

[0071 ] The headings used herein are for organizational purposes only and are not meant to be used to limit the scope of the description or the claims. As used throughout this application, the word "may" is used in a permissive sense (i.e., meaning having the potential to), rather than the mandatory sense (i.e., meaning must). Similarly, the words “include”,“including”, and“includes” mean including but not limited to. To facilitate understanding, like reference numerals have been used, where possible, to designate like elements common to the figures.

Detailed Description

[0072] The description set forth below is intended as a description of various

embodiments of the described subject matter and is not necessarily intended to represent the only embodiment(s). In certain instances, the description includes specific details for the purpose of providing an understanding of the described subject matter. Flowever, it will be apparent to those skilled in the art that embodiments may be practiced without the specific details as described.

[0073] Any reference in the specification to“one embodiment” or“an embodiment” or “another embodiment” means that a particular feature, structure, characteristic, operation, or function described in connection with an embodiment is included in at least one embodiment. Thus, any appearance of the phrases“in one embodiment” or“in an embodiment” in the specification is not necessarily referring to the same embodiment. Further, the particular features, structures, characteristics, operations, or functions may be combined in any suitable manner in one or more embodiments, and it is intended that embodiments of the described subject matter can and do cover modifications and variations of the described embodiments.

[0074] The phrases“at least one”,“one or more”, and“and/or” are open-ended expressions that are both conjunctive and disjunctive in operation. For example, each of the expressions“at least one of A, B and C”,“at least one of A, B, or C”,“one or more of A, B, and C”,“one or more of A, B, or C” and“A, B, and/or C” means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B, and C together.

[0075] The term“a” or“an” entity refers to one or more of that entity. As such, the terms“a” (or“an”),“one or more” and“at least one” can be used interchangeably herein. It is also to be noted that the terms“comprising”,“including”, and“having” can be used interchangeably.

[0076] A compound of this disclosure includes those described generally, and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version,“Handbook of Chemistry and Physics”, 75th Ed., CRC Press, New York, NY (1995). Additionally, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito, CA (1999), and "March's Advanced Organic Chemistry", 5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York, NY (2001 ), the entire contents of which are hereby incorporated by reference.

[0077] The term“hydrocarbon”, used herein refers to paraffinic and naphthenic compounds, or any mixtures of paraffin, naphthenic, or paraffin and naphthenic compounds. Paraffinic compounds may either be linear (n-paraffins) or branched (i- paraffins). Examples of linear paraffins are pentane, hexane, heptane etc. Examples of branched paraffins are isooctane, isobutane, isopentane etc. Naphthenic compounds are cyclic saturated hydrocarbons, i.e. cycloparaffins. Such hydrocarbons with cyclic structure are typically derived from cyclopentane or cyclohexane. A naphthenic compound may comprise a single ring structure (mononaphthene) or two isolated ring structures (isolated dinaphthene), or two fused ring structures (fused dinaphthene) or three or more fused ring structures (polycyclic naphthenes or polynaphthenes).

[0078] The term "hydrocarbon solvent" refers to one or more hydrocarbons which have solvency for mineral oil. Typically, the hydrocarbon solvent comprises at least one of normal or branched chain paraffins or olefins, cyclic hydrocarbons and aromatic hydrocarbons. Often, the hydrocarbon solvent is comprised of at least 50 wt. %, preferably at least 75 wt. % and most preferably at least 90 wt. % of normal or branched chain paraffins or olefins based on the weight of the hydrocarbon solvent. In some embodiments, the hydrocarbon solvent is selected from the group consisting of isoparaffins and normal paraffins. In other embodiments, the hydrocarbon solvent is a normal paraffin. In one embodiment, the hydrocarbon solvent is petroleum ether. In still other embodiments, the hydrocarbon solvent comprises from 5 to 15 carbon atoms per molecule. In other embodiments, the hydrocarbon solvent comprises 7 to 10 carbon atoms per molecule. In addition, the hydrocarbon solvent does not require the presence of functional groups such as, for example, esters, alcohols or adds. In another embodiment, the hydrocarbon solvent is a mixture of petroleum ether and ethyl acetate. In yet other embodiments, that the hydrocarbon solvent contain less than about 5 wt. % and more preferably less than about 1 wt. % of oxygen-containing functional groups such as, for examples, esters, alcohols, acids, or mixtures thereof, based on the weight of the hydrocarbon solvent.

[0079] The term“hydrocarbyl” refers to a monovalent moiety formed by removing a hydrogen atom from a hydrocarbon. The term‘hydrocarbyl’ includes alkyl groups, alkenyl groups, and alkynyl groups. A preferred“hydrocarbyl” group is an“alkyl” group. Representative hydrocarbyl groups are alkyl groups having 1 to 25 carbon atoms, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, undecyl, decyl, dodecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, and tricosyl, and the isomeric forms thereof such as iso-propyl, t-butyl, iso-butyl, sec-butyl, 1 ,1 -dimethylpropyl, 1 ,2- dimethylpropyl, 2,2-dimethylpropyl, 1 -ethylpropyl, 1 -methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1 ,1-dimethylbutyl, 1 ,2-dimethylbutyl, 1 ,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethyl-butyl; alkenyl groups having 2 to 25 carbon atoms, such as methenyl, ethenyl, 1 -propenyl, 2-propenyl, iso-propenyl, 1 - butenyl, 2-butenyl, 3-butenyl, iso-butenyl, sec-butenyl, 1 -pentenyl, 2-pentenyl, 3- pentenyl, 4-pentenyl, hexenyl, heptenyl, octenyl and the isomeric forms thereof; alkynyl groups having 2 to 25 carbon atoms, such as ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1 -pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl, pentynyl, and octynyl, and the isomeric forms thereof. A hydrocarbyl group may also be substituted with a“cyclohydrocarbyl” group. Accordingly, groups such as 2- (cyclopropyl)-ethyl, cyclohexylmethyl, cyclopropylethyl, and cyclopropylmethyl, are contemplated hydrocarbyl groups.

[0080] In some embodiments, a“hydrocarbyl group” contains 1 to 6 members (C Ce), or for alkenyl or alkynyl groups 2 to 6 members (C2-C6). In other embodiments, the hydrocarbyl radical contains 1 to 3 members (C1-C3), or for alkenyl or alkynyl groups 2 to 3 members (C2-C3). In yet other embodiments, the hydrocarbyl radical may contain from 1 to 17 substitutions, or in another embodiment from 1 to 5 substitutions. The hydrocarbyl group may also contain one or more substitutions.

[0081 ] The term“cyclohydrocarbyl”, by itself or part of another substituent, unless otherwise stated, refers to a cyclic hydrocarbyl group which may be fully saturated, monounsaturated, or polyunsaturated but not aromatic, and includes C3-C15 carbon atoms in a ring system. The cyclohydrocarbyl group may contain one or more

substitutions. In one embodiment, the ring contains 3 to 6 members (C3-C6).

[0082] In another embodiment, a cyclohydrocarbyl group may have from 1 to 11 substitutions, or in another embodiment from 2 to 6 substitutions. Examples of cyclohydrocarbyl groups include, but are not limited to cyclopropyl, cyclopentyl, cyclohexyl, cyclohex-1 -enyl, cyclohex-3-enyl, cycloheptyl, cyclooctyl, norbornyl, decalinyl, adamant-1 -yl, adamant-2-yl, bicyclo[2.1.0]pentyl, bicyclo[3.1 .0]-hexyl, spiro[2.4]heptyl, spiro[2.5]octyl, bicyclo-[5.1.0]octyl, spiro[2.6]nonyl, bicyclo[2.2.0]hexyl, spiro[3.3]heptyl, bicyclo[4.2.0]octyl, and spiro[3.5]nonyl, and the like.

[0083] The term "alkyl", by itself or as part of another substituent, unless otherwise stated, refers to a straight chain or branched chain, or or combination thereof, having the number of not more than 15 carbon atoms. Examples of saturated hydrocarbon radicals include, but are not limited to groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t- butyl, isobutyl, sec-butyl, cyclopropyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, and dodecyl, 2-(cyclopropyl)ethyl, cyclohexylmethyl, cyclopropylethyl, cyclohexyl, cyclopropylmethyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1 ,1 - dimethylpropyl, 1 ,2- dimethylpropyl, 2,2-dimethylpropyl, 1 -ethylpropyl, 1 - methylpentyl, 2- methylpentyl, 3- methylpentyl, 4-methylpentyl, 1 ,1 - dimethylbutyl, 1 ,2- dimethylbutyl, 1 ,3- dimethylbutyl, 2,2- dimethylbutyl, 2,3- dimethylbutyl, 3,3-dimethylbutyl, ethylmethylpropyl,

trimethylpropyl, methylhexyl, dimethylpentyl, ethylpentyl, ethylmethylbutyl, dimethylbutyl, spiropentyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to vinyl, prop-2-enyl, crotyl, isopent-2-enyl, butadien-2-yl, penta-2,4-dienyl, penta-1 ,4-dien-3-yl, ethynyl, prop-1 -ynyl, prop-3-ynyl, but-3-ynyl, and the higher homologs and isomers, and the like.

[0084] The term "alkenyl", by itself or as part of another substituent, unless otherwise stated, refers to a straight chain or branched chain radical, or combination thereof, which may be monounsaturated or polyunsaturated and can include divalent and multivalent radicals, having the number of not more than 15 carbon atoms. The alkenyl radicals may contain one or more substituents. In one embodiment, the alkenyl radical contains 2 to 5 members (C2-C5). In another embodiment, the radical contains from 1 to 12 substitutions, or in another embodiment from 2 to 6 substitutions.

[0085] The term "alkynyl", by itself or as part of another substituent, unless otherwise stated, refers to a straight chain or branched chain radical, or combination thereof, which contains one or more triple bond and can include divalent and multivalent radicals, having the number of not more than 15 carbon atoms. The alkynyl radicals may contain one or more substituents. In one embodiment, the alkynyl radical contains 2 to 5 members (C2-C5). In another embodiment, the radical contains from 1 to 12

substitutions, or in another embodiment from 2 to 6 substitutions.

[0086] The term "aryl", unless otherwise stated, used alone or as part of a larger moiety as in“arylalkyl”, is an aromatic cyclohydrocarbyl group that is monocyclic or polycyclic containing up to three fused rings, preferably up to two fused rings, and more preferably, monocyclic. Examples of aryl groups include, but are not limited to phenyl, naphthyl, anthracenyl, and phenanthryl and substituted phenyl, naphthyl, anthracenyl, and phenanthryl groups.

[0087] Phenyl, naphthyl, substituted phenyl, and substituted naphthyl groups are preferred aryl groups, with unsubstituted phenyl and substituted phenyl groups being more preferred. In one embodiment, the ring system may have 1 to about 5

substitutions, or in another embodiment 2 to 3 substitutions are present on the ring system. In one embodiment, the ring system has 1 substitution.

[0088] The term“phenyl” as used herein is a C₆H₅ group. The term“phenyl” may be abbreviated herein as“Ph”. “Phenyl” groups may be substituted. Similarly, the term “naphthyl” as used herein is a C10H7 group. “Naphthyl” groups may be substituted.

[0089] The term "heteroalkyl", by itself or as part of another substituent, unless otherwise stated, refers to a straight or branched chain, or cyclic hydrocarbyl radical, or combinations thereof, consisting of one to fourteen carbon atoms and from one to six heteroatoms selected from oxygen, nitrogen, sulfur, and silicon, and where the nitrogen, sulfur and silicon atoms may optionally be oxidized and the nitrogen atom may optionally be quaternized. The heteroatoms O, N and S may be placed at any interior position of the heteroalkyl group. The heteroatom Si may be placed at any position of the heteroalkyl group, including the position at which the heteroalkyl group is attached to the remainder of the molecule. Examples include, but are not limited to 2-methoxyethyl, 2- (methylamino)ethyl, 2-(dimethylamino)ethyl, 2-(ethylthio)methyl, 2-(methylsulfinyl)ethyl, 2-(methylsulfonyl)ethyl, 2-methoxyvinyl, trimethylsilyl, dimethyl(vinyl)silyl, 2- (cyclopropylthio)ethyl, and 2-(methoxyimino)ethyl. Up to two heteroatoms may be consecutive, such as, for example, (methoxyamino)methyl and trimethylsilyloxy.

[0090] The term "heterocyclyl", by itself or as part of another substituent, represents, unless otherwise stated, cyclic version of "heteroalkyl". Additionally, for heterocyclyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of heterocyclyl include, but are not limited to piperidinyl, piperidin-2-yl, piperidin-3-yl, morpholin-4-yl, morpholin-3-yl, tetrahydrofuran- 2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, piperazinyl, piperazin-2-yl, and the like. In one embodiment, the heterocyclyl radical contains 3 to 6 members (C3-C6). In another embodiment, the radical may contain from 1 to 6 substitutions, or in another embodiment from 1 to 5 substitutions.

[0091 ] The terms "alkoxy" refers to those groups attached to the remainder of the molecule via an oxygen atom. Suitable examples of alkoxy groups include, but are not limited to methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, pentoxy, hexoxy, heptoxy, and the like.

[0092] The term "aryl", unless otherwise stated, used alone or as part of a larger moiety as in“arylalkyl” a polyunsaturated, aromatic, hydrocarbyl substituent which can be a monocyclic system or polycyclic ring system (with up to three rings) which are fused together or linked covalently. The monocyclic or polycyclic ring system comprises about 5 to about 16 carbon atoms. Suitable examples of aryl groups include, but are not limited to phenyl, naphthyl, anthracenyl, and the like. The term aryl also refers to rings that may contain one or more substituents. In one embodiment, the ring system can have from 1 to 5 substitutions, or in another embodiment from 2 to 3 substitutions.

[0093] The term "heteroaryl", unless otherwise stated, used alone or as part of a larger or smaller moiety as in“aryl”, contain from one to four heteroatoms selected from nitrogen, oxygen, and sulfur, where the nitrogen and sulfur atoms are optionally oxidized, and one or several nitrogen atoms are optionally quaternized. A heteroaryl group may be attached to the remainder of the molecule through a heteroatom.

[0094] A heteroaryl group may contain one ring or two fused rings. Non-limiting examples of heteroaryl groups include, 1 -pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 1 - imidizoyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4- oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5- thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,

4-pyrimidyl, 5-benzo-thiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1 -isoquinolyl,

5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. In one

embodiment, examples of heteroaryl groups include pyridine, thiophene, thiazole, imidazole, benzimidazole, pyrazole, and oxazole.

[0095] The terms "arylalkyl" and "heteroarylalkyl" is meant to include those radicals in which an aryl or heteroaryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, and the like) including those alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced by, for example, an oxygen atom (e.g.,

phenoxymethyl, pyrid-2-yloxymethyl, 3-(naphth-1 -yloxy)propyl, and the like). The term “benzyl” as used herein referes to a group in which a phenyl group is attached to a CH₂ group (i.e. a CH₂Ph group). The term substituted benzyl refers to a group in which either the CH₂ linker or the phenyl group contains one or more substitutions. In one

embodiment, the phenyl group may have 1 to 5 substitutions, or in another embodiment 2 to 3 substitutions.

[0096] Each of the above terms "hydrocarbyl", "cyclohydrocarbyl", "alkoxy", "aryl", "heteroaryl", "arylalkyl", and "heteroarylalkyl" may be present in substituted and unsubstituted forms of the indicated radical unless otherwise stated. The "hydrocarbyl", "cyclohydrocarbyl", "alkoxy", "aryl", "heteroaryl", "arylalkyl", and "heteroarylalkyl" groups are optionally substituted by one or more groups that may be the same or different and which are, independently, selected from halogen, halohydrocarbyl which may be mono-, partially substituted, or completely substituted (e.g., in the form of CF₃, CF₂CF₃, CHF₂, CH₂F, and the like), R', OR', OH, SH, SR', N0₂, CN, C(0)R', C(0)OR', OC(0)R', CON(R')₂, OC(0)N(R')₂, NH₂, NHR', N(R')₂, NHCOR', NHCOH, NHCONH₂, NHCONHR', NHCON(R')₂, NRCOR', NRCOH, NHC0₂H, NHC0₂R', C0₂R', CO₂H, CHO, CONH₂, CONHR', CON(R')₂, S(0)₂H, S(0)₂R', S0₂NH₂, S(0)H, S(0)R', S0₂NHR', S0₂N(R')₂, NHS(0)₂H, NR'S(0)₂H, NHS(0)₂R', NR'S(0)₂R', and Si(R')₃. A saturated carbon atom of any such "hydrocarbyl", "cyclohydrocarbyl", "alkoxy", "aryl", "heteroaryl", "arylalkyl", or "heteroarylalkyl" groups may be optionally substituted as long as valency allows. For each of the foregoing, each occurrence of R' is, independently, selected from

"hydrocarbyl", "cyclohydrocarbyl", "alkoxy", "aryl", "heteroaryl", "arylalkyl", and

"heteroarylalkyl”.

[0097] As used herein, the term "heteroatom" is meant to include oxygen (O), nitrogen (N), and sulfur (S). The heteroatoms oxygen and nitrogen are preferred.

[0098] The terms "alkoxy" refers to those groups attached to the remainder of the molecule via an oxygen atom. Suitable examples of alkoxy groups include, but are not limited to methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, pentoxy, hexoxy, heptoxy, and the like.

[0099] The term "halogen" or“halo” means fluorine, chlorine, bromine, or iodine. The term "halohydrocarbyl" means a hydrocarbyl as defined above wherein one or more hydrogens is replaced with a halogen. A halohydrocarbyl group is typically a substituted alkyl substituent. Examples of such haloalkyl include chloromethyl, 1 -bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, 1 ,1 ,1 -trifluoroethyl, and the like. A haloalkyl is an alkyl as defined above wherein one or more hydrogen is replaced with a halogen. Thus, a“partially halogenated hydrocarbyl” group means a hydrocarbyl group in which some but not all hydrogens are replaced by a halogen. Illustrative partially fluorinated hydrocarbyl groups include difluoromethyl, 6,6,6-trifluorohexyl, and 2,3,-difluoropropyl. The term“fully halogenated hydrocarbyl” means a hydrocarbyl group wherein each hydrogen has been replaced by a halogen. Examples of such fully halogenated hydrocarbyl groups are perfluorohydrocarbyl groups such as trifluoromethyl, perfluorobutyl, perfluoroisopropyl, and perfluorohexyl. Preferred partially and fully halogenated hydrocarbyl groups are partially and fully fluorinated hydrocarbyl groups and partially and fully chlorinated hydrocarbyl groups.

[00100] The phrases“independently selected”,“independently” and their variants, when used in reference to two or more of the same substituent group are used herein to mean that that two or more groups may be the same or different. In addition, where one or more substituent positions are recited for such substituent(s), the position numbers and recited substituents(s) take precedence over“independently”.

[00101] By“effective” amount of a drug, formulation, active ingredient, or drug, is meant a sufficient amount of an active agent to provide the desired local or systemic effect. A“topically effective” or“therapeutically effective” amount refers to the amount of drug needed to effect the desired biological result. [00102] The term“pathogen” is a biological agent that causes disease or illness to its host subject. One aspect of the present disclosure includes compounds having therapeutic effect against fungal or protozoan pathogens.

[00103] The term“acceptable salt” and“agriculturally acceptable salt” is meant to include a salt of a compound of the disclosure which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the disclosure contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert carrier. Examples of agriculturally acceptable base addition salts include sodium, potassium, zinc, calcium, ammonium, organic amino (such as choline or diethylamine or amino acids such as d-arginine, l-arginine, d-lysine, or l-lysine), or magnesium salt, or a similar salt. When compounds of the disclosure contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic,

monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science 66: 1 -19 (1977)). Certain specific compounds of the disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[00104] The term“pharmaceutically acceptable carrier” refers to any medium that provides the appropriate delivery of an effective amount of the disclosed active agent(s) as defined herein, which does not negatively interfere with the effectiveness of the biological activity of the active agent and that is sufficiently non-toxic to the subject. Representative pharmaceutically acceptable carriers include water, oils, both vegetable and mineral, cream bases, lotion bases, emulsion bases, ointment bases and the like. These bases include suspending agents, thickeners, penetration enhancers, and the like. Additional information concerning carriers can be found in Remington: The Science and Practice of Pharmacy, 21 st Ed., Lippincott, Williams & Wilkins (2005), which is incorporated herein by reference.

[00105] The term“active ingredient,”“active pharmaceutical ingredient,” or simply “active” as used herein refer to a pharmaceutical agent, active ingredient, compound, or substance, compositions, or mixtures thereof, that provide a pharmacological, often beneficial, effect.

[00106] The terms“dosage” or“dose” as used herein denote any form of the active ingredient formulation that contains an amount sufficient to produce a therapeutic effect with a single administration. The dosage form can be administered, for example, lx, 2x, 3x, 4x, 5x, 6x, or even more times per day. One or more dosage form can be administered, for example, for 1 , 2, 3, 4, 5, 6, 7 days, or even longer. One or more dosage forms can be administered, for example, for 1 , 2, 3, 4 weeks, or even longer. One or more dosage forms can be administered, for example, for 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12 months, 1 year, 2, years, 3 years, 4 years, 5 years, over 5 years, a decade, multiple decades, or even longer. One or more dosage forms can be administered at a regular interval until the subject or subject in need thereof, does not require treatment, prophylaxis, or amelioration of any disease or condition including a pathogenic infection stemming from a pathogenic microorganism.

[00107] The term“patient” or“subject” refers to an animal, such as a mammal, including a human. The active ingredients and pharmaceutical compositions described herein can be administered, for example, to an animal patient, including, humans and livestock, including but not limited to horses, cows, pigs, sheep, goats, rabbits, dogs, cats, and poultry, including, but not limited to chickens, turkeys, ducks and pet birds In some aspects of the disclosure, the subject is a companion mammal, for example including cats, dogs and other domesticated mammals. In some aspects of the disclosure, the subject is a human. The subject may be of any age or stage of development. For example, a human subject may be a child ( e.g ., ~0-9 years old) or an adolescent (e.g., -10-17 years old) or an adult (e.g., ³18 years of age).

[00108] In some embodiments, especially related to infections by

Cryptosporidium, a subject may include young ruminant livestock, including cattle and neonatal calves, and goats, swine, sheep, dogs, and cats. In particular, infection by C. parvum, C. bovis, C. ryanae, and C. andersoni are embodiments of the present disclosure. [00109] As used herein, all percentages (%) refer to weight percent unless noted otherwise.

[001 10] “Biological medium,” as used herein refers to both in vitro and in vivo biological milieus. Exemplary in vitro“biological media” include, but are not limited to, cell culture, tissue culture, homogenates, plasma, and blood. In vivo applications can be performed in animals and humans.

[001 1 1] Chemical structures represented herein can be determined by those of skill in the art.

[001 12] The presence of one or more possible asymmetric carbon atoms in a compound of the disclosure means that the compounds may occur in chiral isomeric forms, i.e., enantiomeric or diastereomeric forms. Also, atropisomers may occur as a result of restricted rotation about a single bond. The disclosed compounds are intended to include all those possible isomeric forms and mixtures thereof. The present disclosure includes all those possible isomeric forms and mixtures thereof for compounds of the disclosed disclosure. The disclosed compounds are intended to include all possible tautomers (e.g. keto-enol tautomerism) where present. Accordingly, the present disclosure includes all possible tautomeric forms for the disclosed compounds.

[001 13] As described herein, the compounds of the present disclosure are novel and potent compounds against pathogenic microorganisms. In some aspects of the disclosure the pathogenic microorganisms include fungi and protozoans.

[001 14] In particular, it is believed that the benzoxaborole compounds of the present disclosure may perturb, including inhibit, key enzymes involved in protein prenylation and ergosterol synthesis. These enzymes include

geranylgeranyltransferase-l (also known as GGTase-l and CDC-43) and farnesyl diphosphate synthase (also known as Erg20), and farnesyltransferase (Ftase).

[001 15] Without being bound by any theory, it is believed that the anti-fungal and anti-protozoan activity of the compounds disclosed herein is attributed at least in part by these enzyme inhibitor functions.

[001 16] Protein geranylgeranyltransferase-l catalyzes the transfer of a 20-carbon isoprenoid lipid to the sulfur of a cysteine residue located near the C terminus of numerous cellular proteins, including members of the Rho superfamily of small GTPases and other essential signal transduction proteins. The function of GGTase-l requires Zn²⁺ for catalysis, where zinc coordinates the cysteine residue of the protein for catalysis of the isoprenoid substrate. These prenylated CaaX proteins play critical roles in a wide variety of cellular processes, including transmembrane signaling, membrane trafficking, and nuclear events. The homologous enzyme farnesyltransferase plays similar roles by the transfer of a 15-carbon isoprenoid modification.

[001 17] Present GGTase-l and FTase inhibitors are either isoprenoid analogs or peptidomimetics, which compete with CaaX proteins, or bi-substrate inhibitors, which have both isoprenoid and peptidomimetic portions. However, the design of such inhibitors has been difficult owing to the nanomolar binding affinity of native substrate proteins. In addition, these inhibitors are large and often hydrophobic, which leads to lower water-solubility and bioavailability. In addition, these compounds have a poor permeability through fungal cell walls. In contrast, the benzoxaborole compounds described herein are small and ligand efficient with favorable drug properties. As such, one hypothesis is that the present compounds should be readily permeable through fungi membrane and cell walls.

[001 18] Utilizing molecular docking techniques it was discovered that the benzoxaborole compounds described herein occupy the CaaX peptide binding site and the boron group interacts with the Zn ion in the tetrahedral form in the binding pocket of GGTase-l and is believed to perturb, such as an inhibitor, either directly or indirectly, for Erg 20. See, Figures 3 and 4.

[001 19] Pathogenic microorganisms, such as protozoans and fungi, require these enzymes to provide numerous prenylated proteins and ergosterol that are essential for growth. Therefore, these pathogenic microorganisms are believed to be susceptible to inhibitors of prenylating enzymes and ergosterol synthesis pathway. Accordingly, it is contemplated herein that the benzoxaborole compounds are useful for perturbing, such as inhibiting, prenylating enzymes and ergosterol synthesis, perturbing, such as inhibiting, pathogenic protozoans and fungi, and treating subjects having a protozoan or fungal infection.

[00120] In one aspect of the present disclosure is a method for treating a subject having a pathogenic infection by administering a therapeutically effective amount of the benzoxaborole compound according to the present disclosure. In another aspect is a method useful for inhibiting protein prenylation or ergosterol synthesis in a pathogenic microorganism by contacting the pathogenic microorganism with the benzoxaborole compound according to the present dislcosure. [00121] Another aspect is a method of inhibiting via a pathway, namely a protein prenylation enzyme and ergosterol synthesis enzyme. Enzymes that the benzoxaborole compounds disclosed herein are believed to inhibit include farnesyl diphosphate synthase (Erg20), geranylgeranyl transferase (Cdc 43), and farnesyltransferase, either directly or indirectly.

[00122] In some embodiments, the IC50 of the compound for inhibiting the enzyme contacted with the prenylation enzyme is in the range of about about 0.001 nM to about 10,000 nM, about 0.01 nM to about 1 ,000 nM, about 0.01 nM to about 100 nM, about 0.1 nM to about 50 nM, about 0.1 to about 25 nM, or about 0.1 to about 10 nM or about 0.1 to about 2 nM. Thus, in some embodiments, the enzyme is contacted with an amount of the compound in the range of about 0.001 nM to about 20,000 nM, about 0.01 nM to about 10,000 nM, about 0.01 nM to about 1 ,00 nM, about 0.1 to about 1 ,000 nM, about 0.1 to about 500 nM, about 0.1 nM to about 250 nM, about 0.1 to about 100 nM, 0.1 to about 50 nM, about 0.1 to about 20 nM or about 0.1 to about 10 nM, or about 0.1 to about 5 nM. Methods for determining inhibition of protein prenylation enzymes and corresponding assays are known in the art. See, Kohl et al., PNAS. 91 , pp. 9145-9145 (1994), which is incorporated by reference herein.

[00123] It is understood that when in aqueous media, the compounds according to the present disclosure may be present in a reversible equilibrium with water due the lewis acidic nature of the trigonal planar boron center (e.g. compounds A and B shown below). This dynamic equilibrium may important for the biological activity of the compounds. All compounds in the present disclosure in this dynamic equilibrium are another aspect of the present invention.

[00124] In each case, the compounds of the disclosed disclosure are may be present in free form, as a hydrate, as a salt, as a stereoisomer, as an enantiomer, or a tautomeric form; e.g., as a pharmaceutically or agriculturally acceptable salt form.

[00125] The active compound(s) of the presently disclosed subject matter, or compositions thereof, will generally be used in an amount effective to achieve the intended result, for example in an amount effective to treat or prevent a fungal or protozoan infection. The amount of compound administered will depend upon a variety of factors, including, for example, the particular indication being treated, the mode of administration, whether the desired benefit is prophylactic or therapeutic, the severity of the indication being treated and the age and weight of the patient, the bioavailability of the particular active compound, and the like.

[00126] In some embodiments, the pathogenic microorganism (e.g., a fungi or protozoan) is contacted with a concentration of the compound of about 0.05 ppm to about 50 ppm, about 0.1 ppm to about 20 ppm, about 0.1 to about 15 ppm, about 0.1 to about 10 ppm or about 0.5 ppm to about 5 ppm. In some embodiments, the pathogenic microorganism (e.g., a fungi or protozoan) is contacted with a concentration of the compound of about 0.1 mM to about 20 mM, about 0.1 to about 15 mM, about 0.1 to about 10 mM or about 0.5 mM to about 5 mM.

[00127] Effective dosages can be estimated initially from in vitro assays. For example, an initial dosage for use in animals can be formulated to achieve a circulating blood or serum concentration of active compound that is at or above an IC50 of the particular compound as measured in as in vitro assay, such as the in vitro fungal MIC or MFC and other in vitro assays. Calculating dosages to achieve such circulating blood or serum concentrations taking into account the bioavailability of the particular compound is well within the capabilities of skilled artisans. For guidance, see Fingl & Woodbury, “General Principles,” In: Goodman and Gilman's The Pharmaceutical Basis of

Therapeutics, Chapter 1 , pp. 1-46, latest edition, Pagamonon Press, and the references cited therein, which are incorporated herein by reference. Initial dosages may also be estimated from in vivo data, such as animal models. Animal models useful for testing the efficacy of compounds to treat or prevent the various diseases described above are well-known in the art.

[00128] Dosage amounts will typically be in the range of from about 0.0001 or 0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but can be higher or lower, depending upon, among other factors, the activity of the compound, its bioavailability, the mode of administration, and various factors discussed above. Dosage amount and interval can be adjusted individually to provide plasma levels of the compound(s), which are sufficient to maintain therapeutic or prophylactic effect. In cases of local administration or selective uptake, such as local topical administration, the effective local concentration of active compound(s) cannot be related to plasma concentration. [00129] The compound(s) can be administered once per day, a few or several times per day, or even multiple times per day, depending upon, among other things, the indication being treated and the judgment of the prescribing physician.

[00130] Preferably, the compound(s) will provide therapeutic or prophylactic benefit without causing substantial toxicity. Toxicity of the compound(s) can be determined using standard pharmaceutical procedures. The dose ratio between toxic and therapeutic (or prophylactic) effect is the therapeutic index. Compounds(s) that exhibit high therapeutic indices are preferred.

[00131] In some embodiments of the methods described herein, the compounds according to the present dislcosure and compositions comprising the compounds are effective to inhibit the growth or kill a pathogenic fungi. Non-limiting and exemplary fungi include: one or more members of the phyla of Ascomycota, Oomycota, Basidiomycota, and the subphylum Mucoromycotina.

[00132] In some embodiments, the fungi is one or more of the following pathogenic fungi genera, including the genera and the species thereof herein: Absidia corymbifera, Ajellomyces capsulatus, Ajellomyces dermatitidis, Arthroderma benhamiae, Arthroderma fulvum, Arthroderma gypseum, Arthroderma incurvatum, Arthroderma otae, Arthroderma vanbreuseghemii, Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Blastomyces dermatitidis, Candida albicans, Candida glabrata, Candida guilliennondii, Candida krusei, Candida parapsilosis, Candida tropicalis, Candida pelliculosa, Cladophialophora carrionii, Coccidioides immitis, Cryptococcus neoformans, Cunninghamella sp., Epidermophyton floccosum, Exophiala dermatitidis, Filobasidiella neoformans, Fonsecaea pedrosoi, Fusarium solani, Geotrichum candidum, Histoplasma capsulatum, Hortaea werneckii, Issatschenkia orientalis, Madurella grisae, Malassezia fur fur, Malassezia globosa, Malassezia obtusa, Malassezia pachydermatis, Malassezia restricta, Malassezia slooffiae, Malassezia sympodialis, Microsporum canis,

Microsporum filvum, Microsporum gypseum, Mucor circinelloides, Nectria

haematococca, Paecilomyces variotii, Paracoccidioides brasiliensis, Penicillium marneffei, Pichia anomala, Pichia guilliermnnondii, Pneumocystis carinii,

Pseudallescheria boydii, Rhizopus oryzae, Rhodotorula rubra, Scedosporium

apiospernium, Schizophyllum commune, Sporothrix schenckii, Trichophyton

mentagrophytes, Trichophyton rubrum, Trichophyton verrucosum, Trichophyton violaceum, Trichosporon asahii, Trichosporon cutaneum, Trichosporon inkin, and Trichosporon mucoides. In some embodiments of the methods described herein, the compounds according to the present disclosure and compositions comprising the compounds are effective to treat a disease, disorder or symptom thereof, which is associated with one of the following conditions: Aspergillosis, Blastomycosis,

Candidiasis, Chromomycosis, Coccidioidomycosis, Cryptococcosis, Dermatophytoses, Histoplasmosis, Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis, Paracoccidioidomycosis, Penicillium marneffei, Phaeohyphomycosis, Pneumocyctis pneumonia, Rhinosporidiosis, Sporotrichosis, Trichosporonosis, and Zygomycosis. In some embodiments, the genera and the species thereof include Aspergillosis and Candidiasis. In some embodiments, the fungi is selected from Candida albicans and Aspergillus fumigatus.

[00133] In some embodiments, the compounds according to the present disclosure are administered with one or more additional anti-fungal compounds. The combination of a benzoxaborole compound and the additional anti-fungal agent in some embodiments synergisitically inhibits the growth of or kills a pathogenic fungi. In some embodiments, this synergistic action of the benzoxaborole compound and additional anti-fungal agent results in less total benzoxaborole and/or second anti-fungal agent needed for treatment of the fungal infection.

[00134] Exemplary and non-limiting anti-fungal agents include ergosterol inhibitors (e.g., azoles and allylamines), inhibitors of plasma membrane enzymes (e.g., polyenes), glucan synthase inhibitors (e.g., echinocandins), inhibitors of nucleic acid biosynthesis (e.g., flucytosines), calcineurin inhibitors, leucyl t-RNA synthetase inhibitors, and cell cytoskeleton inhibitors and combinations thereof.

[00135] In some embodiments, the benzoxaborole compound can be combined with an additional antifungal agent including but not limited to amphotericin b, nystatine, clotrimazole, miconazole, fluconazole, ketoconazole, itraconazole, voriconazole, posaconazole, ravuconazole, caspofungin, anidulafungin, micafungin, terbinafine, butenafin, 5-fluorocytosine, mycograb, griseofulvin, tebuconazole, cyproconazole, propiconazole, and 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole. In some embodiments, the additional antifungal agent is one or more of tebuconazole, cyproconazole, propiconazole, fluconozole, or 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole.

[00136] In some embodiments, the benzoxaborole compound can be combined with a lanosterol 14a-demethylase inhibitor. Exemplary and non-limiting lanosterol 14a- demethylase inhibitors include albaconazole, azalanstat, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, cyproconazole, eberconazole, econazole, econazole/triamcinolone, efinaconazole, enilconazole, fenticonazole, fluconazole, flutrimazole, fosfluconazole, hexaconazole, isavuconazonium, isoconazole, itraconazole, ketoconazole, levoketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, posaconazole, pramiconazole, propiconazole, ravuconazole,

sertaconazole, sulconazole, tebuconazole, terconazole, tioconazole, uniconazole, and voriconazole. In some embodiments, the lanosterol 14a-demethylase inhibitor includes fluconazole, fosfluconazole, ketoconazole, oxiconazole, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, eberconazole, econazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, efinaconazole, fluconazole, fosfluconazole, terconazole, hexaconazole, isavuconazole, itraconazole, posaconazole, voriconazole, albaconazole, and ravuconazole. In some particular embodiments, the benzoxaborole compounds described herein are combined with an additional antifungal agent selected from fluconazole, fosfluconazole, ketaconazole, and oxiconazole.

[00137] In some embodiments, the benzoxaborole compound can be combined with a calcineurin inhibitor. Exemplary and non-limiting calcineurin inhibitors include tacrolimus and ciclosporin A.

[00138] In some embodiments, the benzoxaborole compound can be combined with a leucyl t-RNA synthetase inhibitor. Exemplary and non-limiting leucyl t-RNA synthetase inhibitors include tavaborole and 5-chloro-1-hydroxy-3H-2,1 -benzoxaborole

(BAGS).

[00139] In some embodiments, 1 , 2, 3, or more additional compounds can be combined with the benzoxaborole compounds described herein.

[00140] In some embodiments, the benzoxaborole compounds described herein and an additonal antifungal agent are provided in a molar ratio of 1 :20 to about 20:1 . In some embodiments, the ratio is about 1 : 10 to about 10:1 . In some embodiments, the ratio is about 1 :5 to about 5:1 . In some embodiments, the ratio is about 1 :2 to about 2:1. In some embodiments, the ratio of the benzoxaborole compound is about 1 :20, about 1 :15, about 1 :10, about 1 :5, about 1 :3, about 1 :2, about 1 :1 , about 2:1 , about 3:1 , about 5:1 , about 10 :1 , about 15:1 or about 20:1. In some embodiments, the benzoxaborole compound and second antifungal agent are provided in a ratio amount which achieves a synergistic inhibitor or killing effect on a pathogenic fungi. [00141] In some embodiments of the methods described herein, the compounds of the present disclosure and compositions comprising the compounds are effective to inhibit the growth or kill a pathogenic protozoan. Non-limiting and exemplary protozoans include: one or more members of the phylum Sarcomastigophora, Apicomplexa, Microspora, and Ciliophora.

[00142] In some embodiments, the protozoan infection comprises Plasmodium , Trypanosoma, Leishmania, Toxoplasma, Eimeria, Neospora, Cyclospora, Giardia, Entamoeba, Dientamoeba, Naegleria, Acanthamoeba, Babesia, Isospora, Sarcocystis, Enterocytozoon, Balantidium, Pneumocystis, or Cryptosporidium in one embodiment, the infection comprises Plasmodium or Cryptosporidium in some particular

embodiments, the protozoan is a species, which causes malaria. Thus, in some embodiments, the protozoan comprises Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, and Plasmodium malariae.

[00143] In some embodiments, the benzoxaborole compounds are administered with a second anti-protozoan agent. Exemplary and non-limiting protozoal agents include metronidazole, nifurtimox, atovaquone, pentamidine, benznidazole, quinacrine, dehydroemetine, sodium stibogluconate, eflornithine, suramin, emetine, tinidazole, fenbendazole, iodoquinol, melarsoprol, meglumine antimonite, nitazoxanide, chloroquine, hydroxychloroquine, primaquine, mefloquine, quinine, pyrimethamine, and doxycycline.

Pharmaceutical Compositions

[00144] In one aspect, the disclosure provides a pharmaceutical composition comprising a compound of the present disclosure and, in particular, and a

pharmaceutically acceptable carrier.

[00145] In another aspect, the invention provides a kit comprising an effective amount of a compound of the present disclosure, in unit dosage form, together with instructions for administering the compound to a subject suffering from or susceptible to a fungal or protozoan disease or infection.

[00146] In one embodiment, the compounds disclosed herein are in the form of a pharmaceutically-acceptable formulation for administration to a subject. The formulation can be in the form of an immediate release formulation, delayed release formulation, or a sustained release formulation. Delayed release formulations are generally known in the art and are those that release the active ingredient after about 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, or even 24 hours or longer. Similarly, sustained release formulations are known and are those, for example, that provided sustained delivery of the compound to a subject for at least 12 hours, 24 hours, 36 hours, 48 hours, one week, two weeks, three weeks, or four weeks after the pharmaceutically- acceptable formulation is administered to the subject.

[00147] In another embodiment, the compound is administered parenterally or intraperitoneally. The compound can be either in solution, as a dispersion, or as an emulsion. Dispersions can also be prepared, for example, in glycerol, liquid

polyethylene glycols, and mixtures thereof, and in oils.

[00148] Exemplary and non-limiting pharmaceutical carriers are sugars, such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetates; powdered tragacanth; malt; gelatin; talc; stearic acids; magnesium stearate; calcium sulfate; vegetable oils, such as peanut oils, cotton seed oil, sesame oil, olive oil, corn oil and oil of theobroma; polyols such as propylene glycol, glycerine, sorbitol, mannitol, and polyethylene glycol; agar; alginic acids; pyrogen-free water; isotonic saline; and phosphate buffer solution; skim milk powder; as well as other non-toxic compatible substances used in pharmaceutical formulations such as Vitamin C, estrogen and echinacea, for example. Wetting agents and lubricants such as sodium lauryl sulfate, as well as coloring agents, flavoring agents, lubricants, excipients, tableting agents, stabilizers, anti-oxidants and preservatives, can also be present. Solubilizing agents, including for example, cremaphore and beta-cyclodextrins can also used in the pharmaceutical compositions herein.

[00149] Pharmaceutical compositions comprising the active compounds of the presently disclosed subject matter (or prodrugs thereof) can be manufactured by means of conventional mixing, dissolving, granulating, dragee-making levigating, emulsifying, encapsulating, entrapping or lyophilization processes. The compositions can be formulated in a conventional manner using one or more physiologically acceptable carriers, diluents, excipients or auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically.

[00150] Pharmaceutical compositions of the presently disclosed subject matter can take a form suitable for virtually any mode of administration, including, for example, topical, ocular, oral, buccal, systemic, nasal, injection, transdermal, rectal, vaginal, and the like, or a form suitable for administration by inhalation or insufflation. [00151] For topical administration, the active compound(s) or prodrug(s) can be formulated as solutions, gels, ointments, creams, suspensions, and the like.

[00152] Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral, or pulmonary administration. Useful injectable preparations include sterile suspensions, solutions or emulsions of the active compound(s) in aqueous or oily vehicles. The compositions also can contain formulating agents, such as suspending, stabilizing and/or dispersing agents. The formulations for injection can be presented in unit dosage form (e.g., in ampules or in multidose containers) and can contain added preservatives. Alternatively, the injectable formulation can be provided in powder form for reconstitution with a suitable vehicle, including but not limited to sterile pyrogen free water, buffer, dextrose solution, and the like, before use. To this end, the active compound(s) can be dried by any art-known technique, such as lyophilization, and reconstituted prior to use.

[00153] For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art.

[00154] For oral administration, the pharmaceutical compositions can take the form of, for example, lozenges, tablets or capsules (e.g., hard gelatin capsules or soft gelatin capsules) prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g., pregelatinized maize starch,

polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose,

microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate). The tablets can be coated by methods well known in the art with, for example, sugars or enteric coatings. Preparations for oral administration can be suitably formulated to give controlled release of the active compound or prodrug, as is well known.

[00155] Liquid preparations for oral administration can take the form of, for example, elixirs, solutions, syrups or suspensions, or they can be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations can be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid). The preparations also can contain buffer salts, preservatives, flavoring, coloring and sweetening agents as appropriate.

[00156] For buccal administration, the compositions can take the form of tablets or lozenges formulated in a conventional manner.

[00157] For rectal and vaginal routes of administration, the active compound(s) can be formulated as solutions (for retention enemas), suppositories, or ointments containing conventional suppository bases, such as cocoa butter or other glycerides.

[00158] For nasal administration or administration by inhalation or insufflation, the active compound(s) or prodrug(s) can be conveniently delivered in the form of an aerosol spray from pressurized packs or a nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges for use in an inhaler or insufflator (for example capsules and cartridges comprised of gelatin) can be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[00159] For ocular administration, the active compound(s) or prodrug(s) can be formulated as a solution, emulsion, suspension, and the like, suitable for administration to the eye. A variety of vehicles suitable for administering compounds to the eye are known in the art. Specific non-limiting examples are described in U.S. Pat. No.

6,261 ,547; U.S. Pat. No. 6,197,934; U.S. Pat. No. 6,056,950; U.S. Pat. No.

5,800,807; U.S. Pat. No. 5,776,445; U.S. Pat. No. 5,698,219; U.S. Pat. No.

5,521 ,222; U.S. Pat. No. 5,403,841 ; U.S. Pat. No. 5,077,033; U.S. Pat. No.

4,882,150; and U.S. Pat. No. 4,738,851 , each of which is incorporated herein by reference.

[00160] For prolonged delivery, the active compound(s) or prodrug(s) can be formulated as a depot preparation for administration by implantation or intramuscular injection. The active ingredient can be formulated with suitable polymeric or

hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, e.g., as a sparingly soluble salt. Alternatively, transdermal delivery systems manufactured as an adhesive disc or patch which slowly releases the active compound(s) for percutaneous absorption can be used. To this end, permeation enhancers can be used to facilitate transdermal penetration of the active compound(s). Suitable transdermal patches are described in for example, U.S. Pat. No. 5,407,713; U.S. Pat. No. 5,352,456; U.S. Pat. No. 5,332,213; U.S. Pat. No.

5,336,168; U.S. Pat. No. 5,290,561 ; U.S. Pat. No. 5,254,346; U.S. Pat. No.

5,164,189; U.S. Pat. No. 5,163,899; U.S. Pat. No. 5,088,977; U.S. Pat. No.

5,087,240; U.S. Pat. No. 5,008,1 10; and U.S. Pat. No. 4,921 ,475, each of which is incorporated herein by reference.

[00161] Alternatively, other pharmaceutical delivery systems can be employed. Liposomes and emulsions are well-known examples of delivery vehicles that can be used to deliver active compound(s) or prodrug(s). Certain organic solvents such as dimethylsulfoxide (DMSO) also can be employed.

[00162] The pharmaceutical compositions can, if desired, be presented in a pack or dispenser device which can contain one or more unit dosage forms containing the active compound(s). The pack can, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device can be accompanied by instructions for administration.

Examples

Synthesis

[00163] FIG.1 illustrates a, in accordance with an embodiment of the present disclosure; a schematic representation of a synthetic scheme for making the boron containing compounds of the present disclosure.

[00164] The present application hereby incorporates by reference the experimental details of chemical synsthesis described in PCT/US2019/050162. In addition, several examples are herein repeated for specific reference.

[00165] Example 1 : Compound 44

[00166] Isopropyl (7-chloro-1 -hydroxy-1 ,3-dihydrobenzo[c][1 ,2]oxaborol-6- yl)carbamodithioate

[00167] The title compound was prepared by reacting propane-2-thiol with 7- chloro-6-isothiocyanatobenzo[c][1 ,2]oxaborol-1 (3H)-ol using the procedure described above for the synthesis of its ethyl analog. It was obtained as a white solid (89 mg, yield 22%). ¹H NMR (400 MHz, DMSO-d₆): d 1 1.39 (s, 1 H), 9.24 (s, 1 H), 7.47 (d, J = 7.9 Hz, 1 H), 7.40 (d, J = 7.9 Hz, 1 H), 5.02 (s, 2H), 3.94-3.87 (m, 1 H), 1.35 (d, J = 5.7

Hz, 6H) ppm. HPLC purity: 97.9% at 210 nm and 99.1% at 254 nm. MS (M+H)⁺: m/z = 301.9.

[00168] Example 2: Compound 45

[00169] Propyl (7-chloro-1 -hydroxy- 1 ,3-dihydrobenzo[c][1 ,2]oxaborol-6- yl)carbamodithioate

[00170] The title compound was prepared by reacting propane-1 -thiol with 7- chloro-6-isothiocyanatobenzo[c][1 ,2]oxaborol-1 (3H)-ol using the procedure described above for the synthesis of its ethyl analog. It was obtained as a yellow solid (62 mg, yield 15%). ¹H NMR (400 MHz, DMSO-d₆): d 1 1.47 (s, 1 H), 9.25 (s, 1 H), 7.48 (d, J = 8.0 Hz, 1 H), 7.41 (d, J = 8.0 Hz, 1 H), 5.02 (s, 2H), 3.19 (t, J = 6.9 Hz, 2H), 1.67-1.62 (m, 2H), 0.95 (t, J= 7.1 Hz, 3H) ppm. HPLC purity: 98.5% at 210 nm and 99.5% at 254 nm. MS (M+H)⁺: m/z = 301.9.

[00171] Example 3: Compound 53

[00172] 0-(4-Fluorobenzyl) (7-chloro-1 -hydroxy-1 ,3- dihydrobenzo[c][1 ,2]oxaborol-6-yl)carbamothioate

[00173] A solution of 7-chloro-6-isothiocyanatobenzo[c][1 ,2]oxaborol-1 (3H)-ol (300 mg, 1.3 mmol) and (4-fluorophenyl)methanol (0.3 ml_, 2.8 mmol) in DMF (0.5 ml.) were stirred at 80°C for 4 h. Normal work-up procedure described above was used to generate the title compound (59 mg, 13%) as a white solid. ¹HNMR (400 MHz, DMSO-d₆): d 10.94 (br. s, 1 H), 9.19 (s, 1 H), 7.61-7.08 (br. m, 6H), 5.49 (s, 2H), 4.99 (s, 2H) ppm. HPLC purity: 98.7% at 210 nm and 99.5% at 254 nm. MS (M-H)-: m/z = 350.0.

[00174] Additional compounds of the present disclosure include those of Table A, which may be synthesized following the procedures of the cited reference.

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

or a salt, stereoisomer, enantiomer, or tautomer thereof.

[00175] Additional compounds of the present disclosure include those of Table B, which may be synthesized following the procedures of the cited reference.

Table B

or a salt, stereoisomer, enantiomer, or tautomer thereof.

[00176] The present application hereby incorporates by reference the

experimental details of biological materials, methods, and results described in

PCT/US2019/050162.

Biological Materials and Methods

1. Fungal and oomycetal isolates

[00177] The isolates of Aspergillus flavus NRRL 3518 and Rhizoctonia solani NRRL 66082 were obtained from USDA Agricultural Research Service Culture

Collection. For the purpose of these studies, Aspergillus fumigatus NRRL 62427 was used. The collection of Colletotrichum sublineolum FSP270 was gifted by Dr. Louis Prom at USDA-ARS Crop Germplasm Research in College Station, TX. The isolates of Botrytis cinerea B16, Botrytis cinerea B17, Candida albicans was obtained from the Plant Pathology and Environmental Microbiology Department at The Pennsylvania State University, University Park, PA. The Alternaria solani isolate was kindly gifted by Dr. Inga Meadows at The Department of Entomology and Plant Pathology, Mountain Research Station in North Carolina State University, Waynesville, NC. The collection of Mycosphaerella fijiensis 1 1 CR-33 was given by Dr. Jean Ristaino at the Department of Plant Pathology in North Carolina State University, Raleigh, NC. The isolates of Botrytis cinerea B05.10, Fusarium oxysporum f. sp. cubense TR4, and Phytophthora capsici were obtained from the Texas A&M Agrilife Research, College Station, TX.

2. Fungal and oomycetal inoculum preparation

[00178] Unless specified, most of the organisms were maintained on potato dextrose agar (PDA), and spores can be isolated from the cultures after 1 -2 weeks of incubation at room temperature (20-22°C) with 12 hours fluorescent light (Philips, F40LW) and 12hours blacklight (Philips, F40T12) photoperiod. The final concentrations of all inocula were 5^*10⁴ CFU/mL.

[00179] Mycosphaerella fijiensis : Briefly, mycelial cultures of M. fijiensis isolates 1 1 CR-33 grown on PDA medium were macerated in water, and 1-5 ml. of the resulting suspension was pipetted onto plates of modified V8 medium (0.2g/L CaCC>3, 100 mL/L V8 juice and 20g/L Difco agar). Cultures were incubated at 20°C under continuous, cool-white fluorescent and black light. After 5-7 days, sporulation plates were stimulated to produce conidia by adding 2 ml. water and brushing the plates with a paint brush or cell spreader and removing the resulting suspension. After another 5-7 days, conidia were harvested in the same way, adding 2 ml. 0.05% Tween 20 solution, brushing the plates to dislodge spores, and removing the spore suspension by pipetting. Spores were diluted in half strength broth medium.

[00180] Rhizoctonia solanr. due to insufficient spore obtained from these fungi, inocula were prepared as mycelium visible fragments. In brief, fungal mycelium grown on agar media were cut into 1x1 mm pieces and cultured in autoclaved broth medium (such as PDB and V8). After 3-7 days of incubation at 22-24°C, mycelia were harvested by filtering through one layer of Miracloth. The mycelia were homogenated in half strength of broth medium using household blender for 10 seconds and filtered through one layer Miracloth. The resultant visible fragments were diluted in half strength broth medium. [00181] Fusarium oxysporum f. sp. cubense : the isolate of Fusarium oxysporum f. sp. cubense TR4 was maintained on V8 agar (20% - 200 ml. V8 juice, 2 g CaCO3, 15 g Agar, 800 ml. distilled water. Spore suspensions were prepared in half strength PDB broth medium with 0.1% Tween 20.

In vitro antifungal and anti-oomycetal efficacy of boron-based molecules

[00182] Boron-based compounds were stocked in DMSO with the concentration of 5000 mg/mL (stored at -20°C). The stock solutions were further diluted into sterile half strength broth media in the in vitro assay, in which DMSO final concentration is not greater than 1% (v/v).

[00183] The minimal inhibitory concentrations (MICs) for individual compounds were determined by following a modified broth microdilution protocol. The studies were performed in flat bottom, 96-well microtiter plates (Greiner Bio-One).

[00184] The individual MICs were determined in triplicate in a final volume of 0.2 mL/well with antifungal concentrations of 0.2 - 25 mg/mL (8 serial dilutions down from 25 mg/mL [25, 12.5, 6.25, 3.13, 1.56, 0.78, 0.39 and 0.20 mg/mL]; control studies with 0 mg/mL of compounds were performed in parallel for each plate). Plates sealed with clear polyester film (VWR) were incubated at a temperature of about 22°C. The progress of fungal growth was monitored at 72 hours. The MICs were determined as the lowest antifungal concentrations that inhibited fungal growth by greater than 95% (determined as relative absorbance using the Bio-Tek® Synergy™ H1 microplate reader at 600 nm) relative to the corresponding antifungal-free control.

[00185] The results of the antifungal screening are shown in Figure 2. As noted hereinabove, the present application hereby incorporates by reference the results described in PCT/US2019/050162, as well.

[00186] Additional MIC results of compound 44 and 53 tested against wildetype

(WT) and various mutant fungal clones are shown in Figure 5. Figure 5 shows MIC results of compound 44 and 53 against various WT and resistance mutant strains of Saccharomyces cerevisiae. The MIC is indicated in ppm.

Synergistic Combination of Benzoxaboroles with Second Antifungal Agents

[00187] Synergistic effect is the interaction that causes an increase in the effects of one or both drug compounds. To systematically evaluate the synergistic effect of pairwise drug combinations, functional screening assays that probe combination effects in a dose-response matrix assay are commonly used. In the screening assay where a pair of drugs is plated on microwell plates in a dose-response matrix, thus enabling the assessment of drug combination effects at various dose levels. The degree of synergy or antagonism is to be quantified by comparing the combination response against the expected combination response under the assumption of non-interaction calculated using reference models [the Highest single agent (HSA) model, the Loewe additivity model, the Bliss independence model, and the Zero interaction potency (ZIP) model].

[00188] Benzoxaborole compounds according to the disclosure were tested in the ZIP assay, and the synergistic effect was scored with the ZIP model. The degree of a drug combination effect can be readily visualized as a synergy landscape map over the dose matrix. Dose regions that show strong synergy or antagonism can be further evaluated for more detailed analyses and interpretation about clinical feasibility of the combination ( see lanevski et al., Bioinformatics. 2017 Aug 1 ; 33(15): 2413-2415, which is incorporated by reference herein).

Experimental Procedure

Compound combination

Yeast fungal material

[00189] The wild-type S. cerevisiae strain (BY 4741 ) were purchased from American Type Culture Collection (https://www.atcc.org/Products/Ali/201388.aspx). The yeast strain was cultured on the YPDA medium or in YPD broth. For the synergistic assay, the final concentration of yeast cell was 1 x 10⁴ cfu/mL

Microplate Scheme plan

Operation Procedure

[00190] The procedure for testing synergy was carried out according to the following protocol,

[00191] Step I -- Prepare Solution I (Compound 44 and 53 dilution series) for 4 plates per synergy compound: Prepare Solution I (compound dilution series) for 4 plates: 1) label 8 sterilized tubes (enough for 2500 mL) from "A" to "H"; 2) add corresponding ddH20 into each tube; 3) add corresponding DMSO as the "Solvent (DMSO)" column in the Solution I table; 4) add corresponding compound Stock into the tubes as the "Stock needed" column in the table; 5) use multiple channel (11 -channel) pipette to aliquot each dilution series (50 mL per well) from "A" to "H" into corresponding row (A to H- column 1 to 11) in four (4) plates

[00192] Step II - Prepare Solution II (synergic factor dilution series) for 4 plates per synergy compound: Prepare Solution II (synergic factor dilution series) for 4 plates: 1 ) label 11 sterilized tubes (enough for 2000 mL) from "1" to "11"; 2) add amount of ddH20 in column ""ddH20" into each tube; 3) add solvent (ddH20) as the "Solvent " column in the Solution II table; 4) add corresponding Synergic Factor Stock into the tubes as the "Stock needed" column in the table; 5) use multiple channel (8-channel) pipette to aliquot each dilution series (50 mL per well) from "1" to "11" into corresponding column (1 to 11) in four (4) plates

[00193] Step III - Prepare Solution CK for 4 plates per synergy compound:

Prepare control solutions for 4 plates: 1) label one (1) sterilized tube (enough for 5 mL) as control; 2) add ddH20 as showing in the column of "Total ddH20 needed for 4 plates (mL)" into the tube; 3) add DMSO as the column of "Total solvent DMSO needed for 4 plates (mL)"; 4) use multiple channel (8-channel) pipette to aliquot 10OuL Solution CK per well into the 12th column in four (4) plates [00194] Step IV -- Prepare Solution III (microbial spore dilution series) for 4 plates per synergy compound: Prepare Solution III (microbial spore dilution series) for 4 plates: 1) label one (1) sterilized tube (enough for 50 mL); 2) dilute the microbial spore stock using YPDB; 3) use multiple channel (8-channel) pipette to aliquot 100uL Solution III per well into corresponding column (1 to 11) in four (4) plates; 5) use multiple channel pipette (4 channels) to aliquot 100 µL Solution III into Column 12th "A" to "D" wells in 4 plates -- positive control; 6) use multiple channel pipette (4 channels) to aliquot 100 µL 1/2xPDB into Column 12th "E" to "H" wells in 4 plates -- negative control

[00195] Step V -- Culture the plates at 25^oC for 48 hours; Measure OD600 with the plate reader.

Result analysis

[00196] The synergistic effects were observed between compound 44 and DMI compounds (tebuconazole and cyproconazole). Delta values of the combination have been obtained using the SynergyFinder web application. Positive value means synergy was found. More positive, stronger the synergy (see Table C). The ZIP synergy maps showing the positive synergy results are shown in Figures 6 and 7.

[00197] Further testing of synergy showed synergy between compound 53 and 44 with DMI fungicides including tebuconazole, cyprozonazole, and propiconazole. Leucyl- tRNA synthetase inhibitors, including 5-chlorQ-1 -hydroxy-3H-2,1 -benzoxaborole were found to synergize with compound 53

[00198] As shown above, owing to their mechanism of action, the benzoxaborole compounds 53 and 44 did not synergize with SDHI fungicides including boscalid and fluxapyroxad. In addition, these compounds did not exhibit synergy with Qol fungicides including azoxystrobin and pyraclostrobin.

Antiparasite efficacy of boron-based molecules against Plasmodium falciparum

[00199] The most dangerous forms of malaria come from P. falciparum, where most deaths are incurred. Accordingly, the benzoxaborole compounds disclosed herein will be tested against P. falciparum as previously reported for other antimalarial compounds (see Sonoiki et al., Nat. Commun. Mar 6; 8:14574 (2017), which is incorporated by reference herein).

Example A

[00200] Strains of P. falciparum include 3D7, W2, Dd2, K1 , HB3, FCR3 and TM90C2B; obtained from the Malaria Research and Reference Reagent Resource Center. These strains were maintained utilizing standard methodologies at 2-3% hematocrit in Roswell Park Memorial Institute (RPMI)-1640 medium (Invitrogen) supplemented with 0.5% Albumax II (GIBCO), 2mM L-glutamine, 100mM hypoxanthine,

5 mgml_1 gentamicin, 28mM NaHC03 and 25mM HEPES at 37 _C in an atmosphere of 5% 0 , 5% C0₂ and 90% N₂.

[00201] For parasite reduction ratio studies, the parasites were cultured at 2% haematocrit in RPMI-1640 medium supplemented with 0.5% Albumax II, 2% D-sucrose, 0.3% glutamine and 150mM hypoxanthine. In vitro dose-response assays were perfomed according to the following three methodologies.

[00202] First, parasites were synchronized with D-sorbitol in cell culture plates with the benzooxaborale compounds disclosed herein and a standard anti-malarial agent (e.g., chloroquine). DMSO or another vehicle was used as control. After 48 hours of culture, the cultures were fixed with formaldehyde and stained for counting using a flow cytometer or other counting method.

[00203] According to the second method, parasites that are mostly rings were seeded in 96-well culture plates at 1% haematocrit and 0.2% parasitemia. Parasites were subjected to a range of drug concentrations, with final DMSO concentrations 0.2%, for 72 h. Live parasites were stained with a dye (e.g., SYBR Green and 100nM MitoTracker Deep Red) and quantified by flow cytometry analysis or other cell counting methodology.

[00204] In the third method, parasites at 2% haematocrit and 0.5% parasitemia were exposed to a range of drugs for 48 h, and parasitemia was then quantified by [3H]- hypoxanthine incorporation and IC50 values were calculated through nonlinear regression.

[00205] In addition, the In vitro parasite reduction may be calculated as previously described (see Sonoiki et al., Nat. Commun. Mar 6; 8:14574 (2017)). This assay used limiting dilution to quantify the number of parasites that are viable after various durations of treatment. P. falciparum parasites was cultured at 2% haematocrit and 0.5% parasitemia and was incubated with the benzoxaborole compounds disclosed herein for 24 to 120 h. After each time point within that time range, the compound was removed by washing, and parasites serially diluted in culture well plates and allowed to continue growing after drug wash-out. The number of viable parasites after various incubation times with compounds was determined after two to four weeks by measuring [3H]- hypoxanthine incorporation in threefold serial dilutions of cultures.

Example B

[00206] A medium-throughput screening approach was used as an alternative plasmodium experimental, using 96-well plates and standard P. falciparum culture conditions. See, Jensen JB, 2002, In vitro culture of Plasmodium parasites. Doolan DL, ed. Malaria Methods and Protocols. Totowa, NJ: Humana Press, 477-488, herein incorporated by reference with regard to such teaching.

[00207] More specifically, human erythrocytes infected with about 1% parasitemia of ring-stage W2 strain P. falciparum (synchronized with 5% sorbitol) were incubated with test compounds in 96-well plates at 37 °C for 48 h in RPMI-1640 medium, supplemented with 25 mM HEPES) pH 7.4, 0.5% Albumax and 100 mM hypoxanthine under an atmosphere of 3% 02, 5% C02, and 91% N2. Initially two concentrations (1 uM and 5uM) of test compounds were included and later 3 fold dilutions (starting at 10uM to 0.5nM) for hits (>= 50% inhibition at 5 uM) for IC50 determination. Negative controls where the cultures contained no inhibitors were included in each experimental run. After 48 h, the cells were fixed in 2% formaldehyde in phosphate-buffered saline (PBS) and transferred into PBS with 100 mM NH4CI, 0.1% Triton X-100, 1 nM YOYO-1.

[00208] Parasitemia was determined from dot plots (forward scatter versus fluorescence) acquired on a FACSort flow cytometer using CellQuest software (Becton Dickinson). See, Sijwali PS, Rosenthal PJ, 2004, Gene disruption confirms a critical role for the cysteine protease falcipain-2 in hemoglobin hydrolysis by Plasmodium

falciparum. Proc Natl Acad Sci U S A 101 : 4384-9, herein incorporated by reference with regard to such teaching.

[00209] Alternatively, the growth might be measured by using a fluorescent assay measuring SYBR green 1 . See, Rasmussen SA, Ceja FG, Conrad MD, Tumwebaze PK, Byaruhanga O, Katairo T, Nsobya SL, Rosenthal PJ, Cooper RA, 2017, Changing Antimalarial Drug Sensitivities in Uganda. Antimicrob Agents Chemotherapy 61 : e01516- 17dhingra, herein incorporated by reference with regard to such teaching.

[00210] Values of IC₅o were determined by plotting percent growth of parasites (relative to the negative controls) against log compound concentration, and fitting the data using Graph Pad Prism software.

Benzoxaborole compound inhibition of Erg 20

[0021 1] Inhibition of the Erg20 enzyme may be tested using the benzoxaborale compounds disclosed herein using an enzyme coupled continuous fluorescence assay as previously described (see Dozier, J and Distefano, M., Analytical Biochemistry, 421 (1 ), pp. 158-163 (2012), which is incorporated by reference with regard to such assay.

Cryptosporidium parvum Growth Inhbition Assay

[00212] The compounds of the present disclosure were tested in a cell-based high throughput screen for C. parvum growth inhibition following the methods described in Bessoff et al., Drug Repurposing Screen Reveals FDA-Approved Inhibitors of Human HMG-CoA Reductase and Isoprenoid Synthesis That Block Cryptosporidium parvum Growth, Antimicrobial Agents and Chemotherapy, pp. 1804-1814, Volume 57 Number 4, April 2013, herein incorporated by reference with regard to such descripton, with some modification as herein noted.

[00213] First, the number of duplicates was altered from the cited publications. The procedure of the present disclosure included four (4) wells per compound. Second, the procedure of the present disclosure did not use the pin transfer method. Rather, the present procedure included use of a pipette instead. Lastly, the procedure of the present disclosure modified the concentration of the active agent. The present procedure used concentrations indicated in the data table of Table D. [00214] Table D

[00215] While this specification contains many specific implementation details, these should not be construed as limitations on the scope of any invention or on the scope of what may be claimed, but rather as descriptions of features that may be specific to particular implementations of particular inventions. Certain features that are described in this specification in the context of separate implementations can also be implemented in combination in a single implementation. Conversely, various features that are described in the context of a single implementation can also be implemented in multiple implementations separately or in any suitable sub-combination. Moreover, although features may be described above as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a sub-combination or variation of a sub-combinations.

[00216] Particular implementations of the subject matter have been described. Other implementations, alterations, and permutations of the described implementations are within the scope of the following claims as will be apparent to those skilled in the art. For example, the actions recited in the claims can be performed in a different order and still achieve desirable results.

[00217] Accordingly, the above description of example implementations does not define or constrain this disclosure. Other changes, substitutions, and alterations are also possible without departing from the spirit and scope of this disclosure.

[00218] A number of embodiments of the present disclosure have been described. While this specification contains many specific implementation details, the specific implementation details should not be construed as limitations on the scope of any disclosures or of what may be claimed, but rather as descriptions of features specific to particular embodiments of the present disclosure.

[00219] Certain features that are described in this specification in the context of separate embodiments can also be implemented in combination in a single embodiment. Conversely, various features that are described in the context of a single embodiment can also be implemented in combination in multiple embodiments separately or in any suitable sub-combination. Moreover, although features may be described above as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a sub-combination or variation of a sub combination.

[00220] In certain implementations, multitasking and parallel processing may be advantageous. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the claimed disclosure.

Claims

WHAT IS CLAIMED IS:

1. A method for treating a subject having a pathogenic infection stemming from a pathogenic microorganism comprising administering a therapeutically effective amount of a compound of formula (I):

wherein:

Z¹ is oxygen or sulfur;

Y¹ is selected from the group consisting of: oxygen, sulfur, and NH;

substitutions;

R⁶ and R⁷ are independently selected from the group consisting of C1-C6 hydrocarbyl and substituted hydrocarbyl, or R⁶ and R⁷ can be taken together with the nitrogen to which they are attached to form a ring of no more than 8 members; and

R⁸ and R⁹ can be taken together to form a 3 - 6 membered ring,

or a salt, stereoisomer, enantiomer, or tautomer thereof.

2. The method according to claim 1 , wherein the pathogenic infection is selected from a mucosal infection, systemic infection, invasive infection, and a superficial infection.

3. The method according to claims 1 or 2, wherein the pathogenic infection is selected from a fungal infection and a protozoan infection.

4. The method of claim 3, wherein the fungal infection is selected from

Aspergillosis, Blastomycosis, Candidiasis, Chromomycosis, Coccidioidomycosis, Cryptococcosis, Dermatophytoses, Histoplasmosis, Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis, Paracoccidioidomycosis, Penicillium

marneffei, Phaeohyphomycosis, Pneumocystis pneumonia, Rhinosporidiosis, Sporotrichosis, Trichosporonosis, and Zygomycosis.

5. The method according to any one of claims 1 -4, wherein the compound is administered in an amount to perturb a prenylation pathway of the pathogenic microorganism.

6. The method of any one of claims 1 -4, wherein the compound is administered in an amount to perturb the ergosterol biosynthetic pathway of the microorganism.

7. The method of claim 5, wherein the perturbation of the prenylation pathway results in an effect comprising reduction of protein prenylation in the in the pathogenic microorganism.

8. The method of claim 6, wherein the perturbation of the ergosterol biosynthetic pathway results in an effect comprising reduction of ergosterol synthesis in the pathogenic microorganism.

9. The method according to any one of claims 3-8, wherein the compound is administered with a second antifungal agent to treat the fungal infection.

10. The method of claim 9, wherein the second antifungal agent is an inhibitor of lanosterol demethylase.

1 1 . The method of claim 10, wherein the inhibitor of lanosterol demethylase is an imidazole or triazole antifungal agent.

12. The method according to claims 10 or 1 1 , wherein the lanosterol demethylase inhibitor is selected from fluconazole, fosfluconazole, ketoconazole, oxiconazole, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, eberconazole, econazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, efinaconazole, fluconazole, fosfluconazole, terconazole, hexaconazole, isavuconazole, itraconazole, posaconazole, voriconazole, albaconazole, and

ravuconazole.

13. The method of claim 9, wherein the second antifungal agent is an inhibitor of leucyl t-RNA synthetase.

14. The method of claim 13, wherein the inhibitor of leucyl t-RNA synthetase is is tavaborole or 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole.

15. The method according to any one of claims 9-14, wherein the compound and the second antifungal agent are synergistic in inhibiting fungal growth.

16. The method of claim 3, wherein the protozoan infection is from a phylum selected from Sarcomastigophora, Apicomplexa, Microspora, and Ciliophora.

17. The method of claim 3, wherein the protozoan infection is selected from

Plasmodium, Trypanosoma, Leishmania, Toxoplasma, Eimeria, Neospora, Cyclospora, Giardia, Entamoeba, Dientamoeba, Naegleria, Acanthamoeba, Babesia, Isospora, Sarcocystis, Enterocytozoon, Balantidium, Pneumocystis, and Cryptosporidium.

18. The method of claim 3, wherein the protozoan infection causes malaria.

19. The method of claim 18, wherein the malaria is caused by an infection with one or more protozoan organisms selected from P. falciparum, P. vivax, P. ovale, and P. malariae

20. The method according to any one of claims 16-19, wherein the compound is administered with a second anti-protozoan agent to treat the protozoan infection.

21 . The method of claim 20, wherein the second anti-protozoan agent is selected from metronidazole, nifurtimox, atovaquone, pentamidine, benznidazole, quinacrine, dehydroemetine, sodium stibogluconate, eflornithine, suramin, emetine, tinidazole, fenbendazole, iodoquinol, melarsoprol, meglumine antimonite, nitazoxanide, chloroquine, hydroxychloroquine, primaquine, mefloquine, quinine, pyrimethamine, and doxycycline.

22. A method of perturbing protein prenylation or ergosterol synthesis in a

pathogenic microorganism comprising contacting the pathogenic microorganism with a compound of formula (I):

wherein:

Z¹ is oxygen or sulfur;

Y¹ is selected from the group consisting of: oxygen, sulfur, and NH;

substitutions;

each R^1a independently is selected from the group consisting of C₁-C₈ hydrocarbyl, NH₂, OH, NO₂, CN, C₁-C₂ haloalkyl, OR⁶, SR⁶, SOR⁶, SO₂R⁶, NHC(O)OR⁶, NR⁶R⁷, C(O)OR⁶, unsubstituted aryl, C3-C6 cyclohydrocarbyl, fluorine, chlorine, bromine, iodine, heteroaryl, and heterocyclyl; R⁶ and R⁷ are independently selected from the group consisting of C1-C6 hydrocarbyl and substituted hydrocarbyl, or

R⁸ and R⁹ can be taken together to form a 3 - 6 membered ring,

or a salt, stereoisomer, enantiomer, or tautomer thereof.

23. The method according to claim 22, wherein the pathogenic microorganism is a fungi or a protozoan.

24. The method according to claims 22 or 23, wherein the pathogenic microorganism is contacted with an amount of the compound to perturb a prenylation pathway of the pathogenic microorganism.

25. The method according to claims 22 or 23, wherein the pathogenic microorganism is contacted with an amount of the compound to perturb a ergosterol biosynthetic pathway of the pathogenic microorganism.

26. The method according to claims 22 or 23, wherein the compound preturbs a prenylation enzyme comprising farnesyl diphosphate synthase (Erg20), geranylgeranyl transferase (Cdc 43), or farnesyltransferase or combinations thereof.

27. The method according to claim 26, wherein the compound preturbs farnesyl diphosphate synthase (Erg20).

28. The method according to claim 26, wherein the compound preturbs

geranylgeranyl transferase (Cdc 43).

29. The method according to claim 26, wherein the compound preturbs

farnesyltransferase.

30. The method according to any one of claims 22-29, wherein a prenylation enzyme has one or more mutations.

31 . The method according to any one of claims 22-30, wherein the pathogenic microorganism is contacted with a concentration of the compound of about 0.05 ppm to about 50 ppm.

32. The method according to any one of claims 23-33, wherein the pathogenic microorganism is a fungi and is further contacted with a second antifungal agent.

33. The method of claim 32, wherein the compound and second antifungal agent is provided in a molar ratio of about 1 :20 to about 20:1.

34. The method of claims 32 or 33, wherein the second antifungal agent is a lanosterol demethylase inhibitor selected from fluconazole, fosfluconazole,

ketoconazole, oxiconazole, bifonazole, butoconazole, chlormidazole, clotrimazole, croconazole, eberconazole, econazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, luliconazole, miconazole, neticonazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, efinaconazole, fluconazole, fosfluconazole, terconazole, hexaconazole, isavuconazole, itraconazole, posaconazole, voriconazole, albaconazole, and ravuconazole.

35. The method of claim 32 or 33, wherein the second antifungal agent is an inhibitor of leucyl t-RNA synthetase.

36. The method of claim 35, wherein the inhibitor of leucyl t-RNA synthetase is is tavaborole or 5-ch loro- 1 -hydroxy-3 H-2 , 1 -benzoxaborole.

37. The method of claim 1 , wherein the pathogenic infection is selected from a fungal infection.

38. The method of claim 37, wherein Y¹ is O or S.

39. The method of claim 37 or 38, wherein Z¹ is S.

40. The method of claim 39, wherein the compound is a compound of formula (II):

41. The method of claim 40, wherein X¹ is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, unsubstituted C1-C3 hydrocarbyl, and C1-C3 hydrocarbyl having 1 -7 halogen substitutions.

42. The method of claim 40 or 41 , wherein X^1’ is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, unsubstituted C1-C3 hydrocarbyl, and C1-C3 hydrocarbyl having 1 -7 halogen substitutions.

43. The method of any one of claims 40 - 42, wherein:

when X¹ is hydrogen, X^1’ is other than hydrogen; and

when X^1’ is hydrogen, X¹ is other than hydrogen.

44. The method of any one of claims 37 - 43, wherein R¹ is selected from the group consisting of: hydrogen, unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C3-C6 cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^{1 a} substitutions, unsubstituted benzyl, benzyl having 1 -7 R^1a substitutions,

unsubstituted heterocyclyl, and heterocyclyl substituted with one or more R^1a substitutions; and

each R^1a independently is selected from the group consisting of C₁-C₈ hydrocarbyl, NH₂, OH, OR⁶, SR⁶, NHC(O)OR⁶, NR⁶R⁷, C(O)OR⁶, unsubstituted aryl, C₃-C₆

cyclohydrocarbyl, fluorine, chlorine, bromine, and iodine.

45. The method of claim 37, wherein the compound is selected from a compound in

Table A:

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

Table A

or a salt, stereoisomer, enantiomer, or tautomer thereof.

46. The method of claim 1 , wherein the pathogenic infection is a protozoan infection.

47. The method of claim 46, wherein the compound is a compound of Formula (III)

48. The method of claim 47, wherein

X¹ is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, unsubstituted C1-C3 hydrocarbyl, and C1 -C3 hydrocarbyl having 1-7 halogen substitutions; and

R¹ is selected from the group consistng of unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1-17 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^1a substitutions, unsubstituted benzyl, and benzyl having 1 -7 R^1a substitutions.

49. The method of claim 46, wherein the compound is selected from Table B:

Table B

or a salt, stereoisomer, enantiomer, or tautomer thereof.

50. The method of any one of claims 46 - 49, wherein the protozoan infection is selected from Plasmodium, Trypanosoma, Leishmania, Toxoplasma, Eimeria, Neospora, Cyclospora, Giardia, Entamoeba, Dientamoeba, Naegleria, Acanthamoeba, Babesia, Isospora, Sarcocystis, Enterocytozoon, Balantidium,

Pneumocystis, and Cryptosporidium.

51 . The method of claim 50, wherein the protozoan infection causes malaria and the malaria is caused by an infection with one or more protozoan organisms selected from P. falciparum, P. vivax, P. ovale, and P. m atari ae.

52. The method of any one of claims 46 - 51 , wherein the pathogenic microorganism is contacted with a concentration of the combination of about 0.05 ppm to about 50 ppm.

53. A combination comprising

a) a compound of formula (I):

wherein:

X¹ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C1-C3 hydrocarbyl, C1-C3 hydrocarbyl having 1-7 halogen substitutions, and cyclopropyl;

X^1’ is selected from the group consisting of: hydrogen, fluorine, chlorine, bromine, CN, unsubstituted C1-C3 hydrocarbyl, C1-C3 hydrocarbyl having 1-7 halogen substitutions, and cyclopropyl;

Z¹ is oxygen or sulfur;

Y¹ is selected from the group consisting of: oxygen, sulfur, and NH;

R¹ is selected from the group consisting of: hydrogen, unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C3-C6 cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted aryl, aryl having 1 -5 R^1a substitutions, unsubstituted benzyl, benzyl having 1 -7 R^1a substitutions, unsubstituted heteroaryl, heteroaryl substituted with one or more R^1a substitutions, unsubstituted heterocyclyl, and heterocyclyl substituted with one or more R^1a substitutions;

each R^1a independently is selected from the group consisting of C₁-C₈ hydrocarbyl, NH₂, OH, N0₂, CN, C₁-C₂ haloalkyl, OR⁶, SR⁶, SOR⁶, S0₂R⁶, NHC(O)OR⁶, NR⁶R⁷, C(O)OR⁶, unsubstituted aryl, C3-C6 cyclohydrocarbyl, fluorine, chlorine, bromine, iodine, heteroaryl, and heterocyclyl;

each of R⁸ and R⁹ independently is selected from the group consisting of:

hydrogen, methyl, ethyl; or

R⁸ and R⁹ can be taken together to form a 3 - 6 membered ring,

or a salt, stereoisomer, enantiomer, or tautomer thereof; and

b) one or more additional agent selected from:

i) a demethylation inhibitor; and

ii) a leucyl-tRNA synthetase inhibitor.

54. The combination of claim 53, wherein the compound of formua (I) is a compound of formula (IV):

55. The combination of claim 54, wherein X¹ is halogen.

58. The combination of claim 53 or 54, wherein Y¹ is S.

57 The combination of claim 53 or 54, wherein Y¹ is O.

58. The combination of any one of claims 53 57, wherein R¹ is selected from the group consisting of: unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C3-C6 cyclohydrocarbyl, C3-C6 cyclohydrocarbyl having 1 -11 R^{1 a} substitutions, unsubstituted aryl, aryl having 1 -5 R^1a substitutions, unsubstituted benzyl, and benzyl having 1 -7 R^1a substitutions.

59. The combination of claim 58, wherein R¹ is selected from the group consisting of: unsubstituted C₁-C₈ hydrocarbyl, C₁-C₈ hydrocarbyl having 1 -17 R^1a substitutions, unsubstituted C₆ cyclohydrocarbyl, C₆ cyclohydrocarbyl having 1 -1 1 R^1a substitutions, unsubstituted phenyl, phenyl having 1-5 R^1a substitutions, unsubstituted benzyl, and benzyl having 1 -7 R^1a substitutions.

60. The combination of any one of claims 53 - 59, wherein the compound is

, or a salt, stereoisomer, enantiomer, or tautomer thereof.

61 . The method of any one of claims 53 - 59, wherein the compound is

salt, stereoisomer, enantiomer, or tautomer thereof.

62. The combination of any one of claims 53 - 61 , wherein the compound and additional agent is provided in a molar ratio of about 1 :20 to about 20:1 .

63. The combination of any one of claims 53 - 62, wherein the demethylation inhibitor is one or more of tebuconazole, cyproconazole, propiconazole and fluconazole.

64. The combination of any one of claims 53 - 62, wherein the leucyl-tRNA synthetase inhibitor is is 5-chioro-T-hydroxy-3H-2,1 -benzoxaborole.

65. The combination of claim 60, comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) tebuconazole.

66. The combination of claim 60, comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and b) cyproconazole.

67. The combination of claim 60, comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) propiconazole.

68. The combination of claim 60, comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) fluconazole.

69. The combination of claim 60, comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) 5-chloro-1 -hydroxy-3H~2,1 -benzoxaboroie.

70. The combination of claim 61 , comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) tebuconazole.

71 . The combination of claim 61 , comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) cyproconazole.

72. The combination of claim 61 , comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) propiconazole.

73. The combination of claim 61 , comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) fluconazole.

74. The combination of claim 61 , comprising:

salt, stereoisomer, enantiomer, or tautomer thereof; and

b) 5-chloro-1 -hydroxy-3H-2,1 -benzoxaborole.

75. A method for treating a subject having a pathogenic infection stemming from a pathogenic microorganism comprising administering a therapeutically effective amount of a combination of any one of claims 53 - 74.

76. The method of claim 75, wherein the pathogenic infection is selected from a mucosal infection, systemic infection, invasive infection, and a superficial infection.

77. The method of claim 75 or 76, wherein the pathogenic infection is a fungal infection.

78. The method of claim 77, wherein the fungal infection is selected from Aspergillosis, Blastomycosis, Candidiasis, Chromomycosis, Coccidioidomycosis, Cryptococcosis, Dermatophytoses, Histoplasmosis, Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis, Paracoccidioidomycosis, Penicillium marneffei, Phaeohyphomycosis, Pneumocystis pneumonia, Rhinosporidiosis, Sporotrichosis, Trichosporonosis, and Zygomycosis.

79. The method of any one of claims 75 - 78, wherein the subject is an animal.

80. The method of claim 79, wherein the subject is a mammal.

81 . The method of claim 80, wherein the mammal is a human.