Carboxamide compounds for the treatment or prevention of malaria and other parasitic infections

Abstract

In accordance with the purpose(s) of the present disclosure, as embodied and broadly described herein, the disclosure, in one aspect, relates to compounds and methods for treating or preventing malaria or other parasitic infections in a subject. In one aspect, the compounds are imidazopyridine and purine carboxamide compounds. In one aspect, a compound having a formula represented by structure I or the pharmaceutically acceptable salt thereof can be used in the methods described herein.

Description

CARBOXAMIDE COMPOUNDS FOR THE TREATMENT OR PREVENTION OF MALARIA AND OTHER PARASITIC INFECTIONSSTATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0001] This invention was made with government support under R01 Al 157445 awarded by the National Institutes of Health. The government has certain rights in the invention.CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to co-pending U. S. Provisional Patent Application No. 63 / 714,920, filed on November 1, 2024, the contents of which are incorporated by reference herein in their entireties.BACKGROUND

[0003] Malaria, a deadly ancient disease that humans have battled over millennia, transmitted by Plasmodium spp. -infected Anopheles mosquitoes, continues to be a major global health threat. According to the World Health Organization, in 2023, the global number of malaria cases rose to 263 million, showing a steady increase since the Covid-19 pandemic.1Despite the availability of various prophylactic and therapeutic antimalarial drugs, the rising incidence of malaria is closely tied to increasing resistance to both artemisinin derivatives2and the partner drugs3 6used in artemisinin-based combination therapy (ACT). Therefore, the development of antimalarials that engage novel drug targets is urgently needed.SUMMARY

[0004] In accordance with the purpose(s) of the present disclosure, as embodied and broadly described herein, the disclosure, in one aspect, relates to compounds and methods for treating or preventing malaria or other parasitic infections in a subject. In one aspect, the compounds are imidazopyridine and purine carboxamide compounds. In one aspect, a compound having a formula represented by structure I or the pharmaceutically acceptable salt thereof can be used in the methods described hereinor a pharmaceutically acceptable salt thereof,wherein— represents a single or double bond;Z is CH or N;L1 is substituted or unsubstituted alkylene;L2 is substituted or unsubstituted alkylene group or substituted or unsubstituted cycloalkylene;Ri is H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocycloalkyl;R2is H or alkyl, or R2and L2together with the N atom to which they are attached form a heterocycloalkyl;R3and R4are independently H, alkyl, or hydroxyalkyl, or R3and R4together with the N atom to which they are attached form a heterocycloalkyl; andYi, Y2, and Y3are independently selected from the group consisting of H, halide, and haloalkyl.

[0005] Other systems, methods, features, and advantages of the present disclosure will be or become apparent to one with skill in the art upon examination of the following drawings and detailed description. It is intended that all such additional systems, methods, features, and advantages be included within this description, be within the scope of the present disclosure, and be protected by the accompanying claims. In addition, all optional and preferred features and modifications of the described embodiments are usable in all aspects of the disclosure taught herein. Furthermore, the individual features of the dependent claims, as well as all optional and preferred features and modifications of the described embodiments are combinable and interchangeable with one another.BRIEF DESCRIPTION OF THE DRAWINGS

[0006] Many aspects of the present disclosure can be better understood with reference to the following drawings. The components in the drawings are not necessarily to scale, emphasis instead being placed upon clearly illustrating the principles of the present disclosure. Moreover, in the drawings, like reference numerals designate corresponding parts throughout the several views.

[0007] FIGS. 1 and 2 show exemplary reaction schemes for synthesizing imidazopyridine analogs.

[0008] FIG. 3 shows an exemplary reaction scheme for synthesizing purine analogs.

[0009] FIG. 4 shows dose-dependent stage susceptibility inhibition profiles for replicating blood stages. Schematic illustration of the experimental timeline (top) depicting each 8-hour exposure interval, along with representative Giemsa-stained images of P. falciparum at specified developmental stages (at the initiation of treatment) used for the drug susceptibility assessment. Dose-response curves of P. falciparum (3D7 strain) exposed for 8 hours at defined developmental stages (early ring, late ring, early trophozoite (trophs), late trophozoites, and schizont) are shown along with their bar plots showing the EC508hvalues when parasites were exposed only during the indicated stage. A 72-hour continuous exposure curve was performed in parallel for each compound as a control. The x-axis for the growthinhibition curves are displayed on a logarithmic scale. Data represent mean ± SEM from two biological replicates performed in technical duplicates.

[0010] FIG. 5 shows parasite reduction ratio (PRR) assay results of compounds 1, 10b, P-3f1, and control compounds performed in P. falciparum 3D7 strain using limiting dilution and lactate dehydrogenase (LDH) activity assay.

[0011] FIGS. 6A-6C show recrudescence of parasitemia following short-term drug exposure in ring stage. (A) Dose-response curves of P. falciparum (NF54 strain, DHA sensitive) exposed for 6, 8 or 72 hours at early ring stage. X-axes are shown on a logarithmic scale. (B) Schematic of the experimental design used to monitor recovery following 6 h of drug exposure in highly synchronous ring-stage cultures. Giemsa-stained thin blood smears were prepared daily to assess parasite morphology and viability, which was independently evaluated by two microscopists under blinded conditions. Graph representing recrudescence outcomes for cultures following treatment with 10b or DHA (control). Only DHA-treated parasites recovered. (C) Giemsa-stained thin blood smears showing a pyknotic form like in those treated with 700 nM DHA or 500 nM 10b after only 6 hours of exposure.

[0012] FIGS. 7A-7B show lack of recrudescence of parasitemia after DHA-induced dormant parasites were exposed to 10b for 6 hours. (A) Dose-response curves of P. falciparum (NF54 strain, DHA sensitive) exposed for 6, 8 or 72 hours at early ring stage. (B) Schematic of the experimental design used to monitor recovery following 6 h of DHA exposure in highly synchronous ring-stage cultures (6-8 h post-invasion) to induce dormancy followed by 6 hours of exposure to 10b at 250 nM or 500 nM. Giemsa-stained thin blood smears were prepared daily to assess parasite morphology and viability, which was independently evaluated by two microscopists under blinded conditions. Graph representing recrudescence outcomes for cultures following treatment with 10b or DHA (control).

[0013] FIGS. 8A-8B show 10b has oral efficacy in P. yoe / / 7-infected mice. Female CF1 mice from Charles River Laboratories were inoculated intravenously with approximately 5.0x104parasitized erythrocytes (murine malaria P. yoelii, Kenya strain MR4 MRA-428) from a donor mouse (experiment day zero). On the following 4 days (experiment days 1-4), solutions of the test compounds in PEG-400 (or PEG-400 only for control mice) were administered by oral gavage once daily. 10b was assessed at 40 and 60 mg / kg / day for 4 days or at 120 and 160 mg / kg as single dose. Parasitemia of each mouse was determined by microscopic examination of Giemsa-stained blood smears on the days indicated in (A).

[0014] Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or can be learned by practice of the invention. The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.DETAILED DESCRIPTION

[0015] Many modifications and other embodiments disclosed herein will come to mind to one skilled in the art to which the disclosed compositions and methods pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the disclosures are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. The skilled artisan will recognize many variants and adaptations of the aspects described herein. These variants and adaptations are intended to be included in the teachings of this disclosure and to be encompassed by the claims herein.

[0016] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0017] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure.

[0018] Any recited method can be carried out in the order of events recited or in any other order that is logically possible. That is, unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0019] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The publications discussed herein are provided solely fortheir disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided herein can be different from the actual publication dates, which can require independent confirmation.

[0020] While aspects of the present disclosure can be described and claimed in a particular statutory class, such as the system statutory class, this is for convenience only and one of skill in the art will understand that each aspect of the present disclosure can be described and claimed in any statutory class.

[0021] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed compositions and methods belong. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly defined herein.

[0022] Prior to describing the various aspects of the present disclosure, the following definitions are provided and should be used unless otherwise indicated. Additional terms may be defined elsewhere in the present disclosure.Definitions

[0023] As used herein, “comprising” is to be interpreted as specifying the presence of the stated features, integers, steps, or components as referred to, but does not preclude the presence or addition of one or more features, integers, steps, or components, or groups thereof. Moreover, each of the terms “by,” “comprising,” “comprises,” “comprised of,” “includes,” “included,” “involving,” “involves,” “involved," and “such as” are used in their open, non-limiting sense and may be used interchangeably. Further, the term “comprising” is intended to include examples and aspects encompassed by the terms “consisting essentially of’ and “consisting of.” Similarly, the term “consisting essentially of’ is intended to include examples encompassed by the term “consisting of.

[0024] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “an excipient" includes, but is not limited to, mixtures or combinations of two or more such excipients, and the like.

[0025] It should be noted that ratios, concentrations, amounts, and other numerical data can be expressed herein in a range format. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about" that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms a further aspect. For example, if the value “about 10” is disclosed, then “10” is also disclosed.

[0026] When a range is expressed, a further aspect includes from the one particular value and / or to the other particular value. For example, where the stated range includes one or both limits, ranges excluding either or both of those included limits are also included in the disclosure, e.g. the phrase “x to y” includes the range from ‘x’ to ‘y’ as well as the range greater than x’ and less than ‘y’. The range can also be expressed as an upper limit, e.g. ‘about x, y, z, or less’ and should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘less than x’, less than y’, and ‘less than z’. Likewise, thephrase ‘about x, y, z, or greater’ should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘greater than x’, greater than y’, and ‘greater than z’. In addition, the phrase “about ‘x’ to ‘y’”, where ‘x’ and ‘y’ are numerical values, includes “about ‘x’ to about ‘y’”.

[0027] It is to be understood that such a range format is used for convenience and brevity, and thus, should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. To illustrate, a numerical range of “about 0.1% to 5%’’ should be interpreted to include not only the explicitly recited values of about 0.1% to about 5%, but also include individual values (e.g., about 1%, about 2%, about 3%, and about 4%) and the sub-ranges (e.g., about 0.5% to about 1.1%; about 5% to about 2.4%; about 0.5% to about 3.2%, and about 0.5% to about 4.4%, and other possible sub-ranges) within the indicated range. Thus, for example, if a component is in an amount of about 1%, 2%, 3%, 4%, or 5%, where any value can be a lower and upper endpoint of a range, then any range is contemplated between 1% and 5% (e.g., 1% to 3%, 2% to 4%, etc.).

[0028] As used herein, the terms “about,” “approximate,” “at or about,” and “substantially” mean that the amount or value in question can be the exact value or a value that provides equivalent results or effects as recited in the claims or taught herein. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art such that equivalent results or effects are obtained. In some circumstances, the value that provides equivalent results or effects cannot be reasonably determined. In such cases, it is generally understood, as used herein, that “about” and “at or about” mean the nominal value indicated ±10% variation unless otherwise indicated or inferred. In general, an amount, size, formulation, parameter or other quantity or characteristic is “about,” “approximate,” or “at or about” whether or not expressly stated to be such. It is understood that where “about,” “approximate,” or “at or about” is used before a quantitative value, the parameter also includes the specific quantitative value itself, unless specifically stated otherwise.

[0029] As used herein, half maximal effective concentration (EC50) is a measure of the concentration of a drug, antibody or toxicant which induces a biological response halfway between the baseline and maximum after a specified exposure time.

[0030] A residue of a chemical species, as used in the specification and concluding claims, refers to the moiety that is the resulting product of the chemical species in a particular reaction scheme or subsequent formulation or chemical product, regardless of whether the moiety is actually obtained from the chemical species. Thus, an ethylene glycol residue in a polyester refers to one or more -OCH2CH2O- units in the polyester, regardless of whether ethylene glycol was used to prepare the polyester. Similarly, a sebacic acid residue in a polyester refers to one or more -CO(CH2)aCO- moieties in the polyester, regardless of whether the residue is obtained by reacting sebacic acid or an ester thereof to obtain the polyester.

[0031] As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. This disclosure is not intended to be limited in any manner by the permissible substituents of organic compounds. Also, the terms “substitution” or “substituted with” include the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. It is also contemplated that, in certain aspects, unless expressly indicated to the contrary, individual substituents can be further optionally substituted ( / .e., further substituted, or unsubstituted).

[0032] The position of a substituent can be defined relative to the positions of other substituents in an aromatic ring. For example, as shown below in relationship to the “R” group, a second substituent can be “ortho," “para," or “meta" to the R group, meaning that the second substituent is bonded to a carbon labeled ortho, para, or meta as indicated below. Combinations of ortho, para, and meta substituents relative to a given group or substituent are also envisioned and should be considered to be disclosed.ortho orthometa metapara

[0033] In defining various terms, “A1,” “A2,” “A3,” and “A4” are used herein as generic symbols to represent various specific substituents. These symbols can be any substituent, not limited to those disclosed herein, and when they are defined to be certain substituents in one instance, they can, in another instance, be defined as some other substituents.

[0034] The term “aliphatic” or “aliphatic group,” as used herein, denotes a hydrocarbon moiety that may be straight-chain ( / .e., unbranched), branched, or cyclic (including fused, bridging, and spirofused polycyclic) and may be completely saturated or may contain one or more units of unsaturation, but which is not aromatic. Unless otherwise specified, aliphatic groups contain 1-20 carbon atoms. Aliphatic groups include, but are not limited to, linear or branched, alkyl, alkenyl, and alkynyl groups, and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0035] The term “alkyl” as used herein is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, / -butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. The alkyl group can be cyclic or acyclic. The alkyl group can be branched or unbranched. The alkyl group can also be substituted or unsubstituted. For example, the alkyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol, as described herein. A “lower alkyl” group is an alkyl group containing from one to six (e.g., from one to four) carbon atoms. The term alkyl group can also be a C1 alkyl, C1-C2 alkyl, C1-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C7 alkyl, C1-C8 alkyl, C1-C9 alkyl, C1-C10 alkyl, and the like up to and including a C1-C24 alkyl.

[0036] Throughout the specification “alkyl” is generally used to refer to both unsubstituted alkyl groups and substituted alkyl groups; however, substituted alkyl groups are also specifically referred to herein by identifying the specific substituent(s) on the alkyl group. For example, the term “halogenated alkyl” or “haloalkyl” specifically refers to an alkyl group that is substituted with one or more halide, e.g., fluorine, chlorine, bromine, or iodine. Alternatively, the term “monohaloalkyl” specifically refers to an alkyl group that is substituted with a single halide, e.g. fluorine, chlorine, bromine, or iodine. The term “polyhaloalkyl” specifically refers to an alkyl group that is independently substituted with two or more halides, i.e. each halide substituent need not be the same halide as another halide substituent, nor do the multiple instances of a halide substituent need to be on the same carbon. The term “alkoxyalkyl” specifically refers to an alkyl group that is substituted with one or more alkoxy groups, as described below. The term “aminoalkyl” specifically refers to an alkyl group that is substituted with one or more amino groups. The term “hydroxyalkyl” specifically refers to an alkyl groupthat is substituted with one or more hydroxy groups. When “alkyl” is used in one instance and a specific term such as “hydroxyalkyl” is used in another, it is not meant to imply that the term “alkyl” does not also refer to specific terms such as “hydroxyalkyl” and the like.

[0037] This practice is also used for other groups described herein. That is, while a term such as “cycloalkyl” refers to both unsubstituted and substituted cycloalkyl moieties, the substituted moieties can, in addition, be specifically identified herein; for example, a particular substituted cycloalkyl can be referred to as, e.g., an “alkylcycloalkyl.” Similarly, a substituted alkoxy can be specifically referred to as, e.g., a “halogenated alkoxy,” a particular substituted alkenyl can be, e.g., an “alkenylalcohol,” and the like. Again, the practice of using a general term, such as “cycloalkyl,” and a specific term, such as “alkylcycloalkyl,” is not meant to imply that the general term does not also include the specific term.

[0038] The term “cycloalkyl” or “cycloalkylene” as used herein is a non-aromatic carbon-based ring composed of at least three carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, and the like. The term “heterocycloalkyl” is a type of cycloalkyl group as defined above, and is included within the meaning of the term “cycloalkyl,” where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. The cycloalkyl group and heterocycloalkyl group can be substituted or unsubstituted. The cycloalkyl group and heterocycloalkyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol as described herein.

[0039] The term “alkylene” as used herein, refers to a divalent saturated aliphatic group, with one or two saturated carbon atom(s) as the point(s) of attachment, a linear or branched, cyclo, cyclic or acyclic structure, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. For example, the alkylene group can be represented by the formula -(CH2)n-, where n is an integer from 1 to 10. The groups, -CH2- (methylene), -CH2CH2-, -CH2C(CH3)2CH2-, and -CH2CH2CH2- are non-limiting examples of alkylene groups. A substituted alkylene group is when one of the hydrogen atoms of the alkylene group is replaced with another group such including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol as described herein.

[0040] The terms “alkoxy” and “alkoxyl” as used herein to refer to an alkyl or cycloalkyl group bonded through an ether linkage; that is, an “alkoxy” group can be defined as — OA1where A1is alkyl or cycloalkyl as defined above. “Alkoxy” also includes polymers of alkoxy groups as just described; that is, an alkoxy can be a polyether such as — OA1— OA2or — OA1— (OA2)a—OA3, where “a” is an integer of from 1 to 200 and A1, A2, and A3are alkyl and / or cycloalkyl groups.

[0041] The term “aryloxy” as used herein to refer to an aryl group bonded through an ether linkage; that is, an “aryloxy” group can be defined as — OA1where A1is aryl as defined herein.

[0042] The term “alkenyl” as used herein is a hydrocarbon group of from 2 to 24 carbon atoms with a structural formula containing at least one carbon-carbon double bond. Asymmetric structures such as (A1A2)C=C(A3A4) are intended to include both the E and Z isomers. This can be presumed in structural formulae herein wherein an asymmetric alkene is present, or it can be explicitly indicated by the bond symbol C=C. The alkenyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol, as described herein.

[0043] The term “cycloalkenyl” as used herein is a non-aromatic carbon-based ring composed of at least three carbon atoms and containing at least one carbon-carbon double bound, i.e., C=C. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, norbornenyl, and the like. The term “heterocycloalkenyl” is a type of cycloalkenyl group as defined above and is included within the meaning of the term “cycloalkenyl,” where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. The cycloalkenyl group and heterocycloalkenyl group can be substituted or unsubstituted. The cycloalkenyl group and heterocycloalkenyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol as described herein.

[0044] The term “alkynyl” as used herein is a hydrocarbon group of 2 to 24 carbon atoms with a structural formula containing at least one carbon-carbon triple bond. The alkynyl group can be unsubstituted or substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol, as described herein.

[0045] The term “cycloalkynyl" as used herein is a non-aromatic carbon-based ring composed of at least seven carbon atoms and containing at least one carbon-carbon triple bound. Examples of cycloalkynyl groups include, but are not limited to, cyclooctynyl, cyclononynyl, and the like. The term “heterocycloalkynyl” is a type of cycloalkenyl group as defined aboveand is included within the meaning of the term “cycloalkynyl,” where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. The cycloalkynyl group and heterocycloalkynyl group can be substituted or unsubstituted. The cycloalkynyl group and heterocycloalkynyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol as described herein.

[0046] The term “aromatic group” as used herein refers to a ring structure having cyclic clouds of delocalized TT electrons above and below the plane of the molecule, where the TT clouds contain (4n+2) TT electrons. A further discussion of aromaticity is found in Morrison and Boyd, Organic Chemistry, (5th Ed., 1987), Chapter 13, entitled “Aromaticity,” pages 477-497, incorporated herein by reference. The term “aromatic group” is inclusive of both aryl and heteroaryl groups.

[0047] The term “aryl” as used herein is a group that contains any carbon-based aromatic group including, but not limited to, benzene, naphthalene, phenyl, biphenyl, anthracene, and the like. The aryl group can be substituted or unsubstituted. The aryl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, — NH2, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol as described herein. The term “biaryl” is a specific type of aryl group and is included in the definition of “aryl.” In addition, the aryl group can be a single ring structure or comprise multiple ring structures that are either fused ring structures or attached via one or more bridging groups such as a carbon-carbon bond. For example, biaryl to two aryl groups that are bound together via a fused ring structure, as in naphthalene, or are attached via one or more carbon-carbon bonds, as in biphenyl. Fused aryl groups including, but not limited to, indene and naphthalene groups are also contemplated.

[0048] The term “aldehyde” as used herein is represented by the formula -C(O)H. Throughout this specification “C(O)” is a shorthand notation for a carbonyl group, i.e., C=O.

[0049] The terms “amine” or “amino” as used herein are represented by the formula — NA1A2, where A1and A2can be, independently, hydrogen or alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. A specific example of amino is — NH2.

[0050] The term “alkylamino” as used herein is represented by the formula — NH(-alkyl) and — N(-alkyl)2, where alkyl is a described herein. Representative examples include, but are notlimited to, methylamino group, ethylamino group, propylamino group, isopropylamino group, butylamino group, isobutylamino group, (sec-butyl)amino group, (ferf-butyl)amino group, pentylamino group, isopentylamino group, (fert-pentyl)amino group, hexylamino group, dimethylamino group, diethylamino group, dipropylamino group, diisopropylamino group, dibutylamino group, diisobutylamino group, di(sec-butyl)amino group, di(ferf-butyl)amino group, dipentylamino group, diisopentylamino group, di(tert-pentyl)amino group, dihexylamino group, A / -ethyl- / V-methylamino group, / V-methyl-ZV-propylamino group, A / -ethyl- / V-propylamino group and the like.

[0051] The term “carboxylic acid” as used herein is represented by the formula — C(O)OH.

[0052] The term “ester” as used herein is represented by the formula — OC(O)A1or — C(O)OA1, where A1can be alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

[0053] The term “ether” as used herein is represented by the formula A1OA2, where A1and A2can be, independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group described herein.

[0054] The terms “halo,” “halogen” or “halide,” as used herein can be used interchangeably and refer to F, Cl, Br, or I.

[0055] The terms “pseudohalide,” “pseudohalogen” or “pseudohalo,” as used herein can be used interchangeably and refer to functional groups that behave substantially similar to halides. Such functional groups include, by way of example, cyano, thiocyanato, azido, trifluoromethyl, trifluoromethoxy, perfluoroalkyl, and perfluoroalkoxy groups.

[0056] The term “heteroalkyl” as used herein refers to an alkyl group containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P and S, wherein the nitrogen, phosphorous and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. Heteroalkyls can be substituted as defined above for alkyl groups.

[0057] The term “heteroaryl” as used herein refers to an aromatic group that has at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus, where N-oxides, sulfur oxides, and dioxides are permissible heteroatom substitutions. The heteroaryl group can be substituted or unsubstituted. The heteroaryl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol as described herein. Heteroaryl groups can be monocyclic, or alternatively fused ring systems. Heteroaryl groups include, but are not limited to, furyl,imidazolyl, pyrimidinyl, tetrazolyl, thienyl, pyridinyl, pyrrolyl, / V-methylpyrrolyl, quinolinyl, isoquinolinyl, pyrazolyl, triazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridazinyl, pyrazinyl, benzofuranyl, benzodioxolyl, benzothiophenyl, indolyl, indazolyl, benzimidazolyl, imidazopyridinyl, pyrazolopyridinyl, and pyrazolopyrimidinyl. Further not limiting examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiophenyl, pyrazolyl, imidazolyl, benzo[c / ]oxazolyl, benzo[c / ]thiazolyl, quinolinyl, quinazolinyl, indazolyl, imidazo[1,2-b]pyridazinyl, imidazo[1,2-a]pyrazinyl, benzo[c][1,2,5]thiadiazolyl, benzo[c][1,2,5]oxadiazolyl, and pyrido[2,3-b]pyrazinyl.

[0058] The terms “heterocycle” or“heterocyclyl,” as used herein can be used interchangeably and refer to single and multi-cyclic aromatic or non-aromatic ring systems in which at least one of the ring members is other than carbon. Thus, the term is inclusive of, but not limited to, “heterocycloalkyl,” “heteroaryl,” “bicyclic heterocycle,” and “polycyclic heterocycle.” Heterocycle includes pyridine, pyrimidine, furan, thiophene, pyrrole, isoxazole, isothiazole, pyrazole, oxazole, thiazole, imidazole, oxazole, including, 1,2,3-oxadiazole, 1,2,5-oxadiazole and 1,3,4-oxadiazole, thiadiazole, including, 1,2,3-thiadiazole, 1,2,5-thiadiazole, and 1,3,4-thiadiazole, triazole, including, 1,2,3-triazole, 1,3,4-triazole, tetrazole, including 1, 2,3,4-tetrazole and 1,2,4,5-tetrazole, pyridazine, pyrazine, triazine, including 1,2,4-triazine and 1,3,5-triazine, tetrazine, including 1,2,4,5-tetrazine, pyrrolidine, piperidine, piperazine, morpholine, azetidine, tetrahydropyran, tetrahydrofuran, dioxane, and the like. The term heterocyclyl group can also be a C2 heterocyclyl, C2-C3 heterocyclyl, C2-C4 heterocyclyl, C2-C5 heterocyclyl, C2-C6 heterocyclyl, C2-C7 heterocyclyl, C2-C8 heterocyclyl, C2-C9 heterocyclyl, C2-C10 heterocyclyl, C2-C11 heterocyclyl, and the like up to and including a C2-C18 heterocyclyl. For example, a C2 heterocyclyl comprises a group which has two carbon atoms and at least one heteroatom, including, but not limited to, aziridinyl, diazetidinyl, dihydrodiazetyl, oxiranyl, thiiranyl, and the like. Alternatively, for example, a C5 heterocyclyl comprises a group which has five carbon atoms and at least one heteroatom, including, but not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, diazepanyl, pyridinyl, and the like. It is understood that a heterocyclyl group may be bound either through a heteroatom in the ring, where chemically possible, or one of carbons comprising the heterocyclyl ring.

[0059] The term “bicyclic heterocycle” or “bicyclic heterocyclyl” as used herein refers to a ring system in which at least one of the ring members is other than carbon. Bicyclic heterocyclyl encompasses ring systems wherein an aromatic ring is fused with another aromatic ring, or wherein an aromatic ring is fused with a non-aromatic ring. Bicyclic heterocyclyl encompasses ring systems wherein a benzene ring is fused to a 5- or a 6-membered ring containing 1, 2 or 3 ring heteroatoms or wherein a pyridine ring is fused to a 5- or a 6-membered ring containing1, 2 or 3 ring heteroatoms. Bicyclic heterocyclic groups include, but are not limited to, indolyl, indazolyl, pyrazolo[1,5-a]pyridinyl, benzofuranyl, quinolinyl, quinoxalinyl, 1,3-benzodioxolyl, 2,3-dihydro-1,4-benzodioxinyl, 3,4-dihydro-2 / - / -chromenyl, 1 / - / -pyrazolo[4,3-c]pyridin-3-yl; 1 / 7-pyrrolo[3,2-b]pyridin-3-yl; and 1H-pyrazolo[3,2-b]pyridin-3-yl.

[0060] The term “heterocycloalkyl” or “heterocycloalkylene” as used herein refers to an aliphatic, partially unsaturated or fully saturated, 3- to 14-membered ring system, including single rings of 3 to 8 atoms and bi- and tricyclic ring systems. The heterocycloalkyl ringsystems include one to four heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein a nitrogen and sulfur heteroatom optionally can be oxidized and a nitrogen heteroatom optionally can be substituted. Representative heterocycloalkyl groups include, but are not limited to, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl.

[0061] The term “hydroxyl” or “hydroxy” as used herein is represented by the formula — OH.

[0062] The term “ketone” as used herein is represented by the formula A1C(O)A2, where A1and A2can be, independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

[0063] The term “azide” or “azido” as used herein is represented by the formula — N3.

[0064] The term “nitro” as used herein is represented by the formula — NO2.

[0065] The term “nitrile” or “cyano” as used herein is represented by the formula — CN.

[0066] The term “silyl” as used herein is represented by the formula — SiA1A2A3, where A1, A2, and A3can be, independently, hydrogen or an alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

[0067] The term “sulfo-oxo” as used herein is represented by the formulas — S(O)A1, — S(O)2A1, — OS(O)2A1, or — OS(O)2OA1, where A1can be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. Throughout this specification “S(O)” is a shorthand notation for S=O. The term “sulfonyl” is used herein to refer to the sulfo-oxo group represented by the formula — S(O)2A1, where A1can be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. The term “sulfone” as used herein is represented by the formula A1S(O)2A2, where A1and A2can be, independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. The term “sulfoxide” as used herein is represented by the formula A1S(O)A2, where A1and A2can be,independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

[0068] The term “thiol” as used herein is represented by the formula -SH.

[0069] “R1,” “R2,” “R3,”... “Rn,” where n is an integer, as used herein can, independently, possess one or more of the groups listed above. For example, if R1is a straight chain alkyl group, one of the hydrogen atoms of the alkyl group can optionally be substituted with a hydroxyl group, an alkoxy group, an alkyl group, a halide, and the like. Depending upon the groups that are selected, a first group can be incorporated within second group or, alternatively, the first group can be pendant ( / .e., attached) to the second group. For example, with the phrase “an alkyl group comprising an amino group,” the amino group can be incorporated within the backbone of the alkyl group. Alternatively, the amino group can be attached to the backbone of the alkyl group. The nature of the group(s) that is (are) selected will determine if the first group is embedded or attached to the second group.

[0070] As described herein, compounds of the invention may contain “optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. In is also contemplated that, in certain aspects, unless expressly indicated to the contrary, individual substituents can be further optionally substituted ( / .e., further substituted, or unsubstituted).

[0071] The term “stable,” as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain aspects, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0072] Suitable monovalent substituents on a substitutable carbon atom of an “optionally substituted” group are independently halogen; -(CH2)o^R°; -(CH2)CMOR°; -0(CH2)o- R°, -O-(CH2)Q^C(O)ORO; -(CH2)O-4CH(OR°)2; -(CH2)O-4SR0; -(CH2)0-4Ph, which may be substituted with R°; -(CH2)o^O(CH2)o--Ph which may be substituted with R°; -CH=CHPh, which may be substituted with R°; -(CH2)0-4O(CH2)0-i -pyridyl which may be substituted with R°; -NO2; -CN; -N3; -(CH2)O_4N(R°)2; -(CH2)O_4N(R0)C(0)R°; -N(R°)C(S)R°;-(CH2)Q^N(R°)C(O)NR°2; -N(R°)C(S)NR°2; -(CH2)0^N(RO)C(O)OR°; -N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; -(CH2)0^C(O)R°; -C(S)R°;-(CH2)O^C(0)OR°; -(CH2)O-4C(0)SR°; -(CH2)0^C(O)OSiR°3; -(CH2)0^OC(O)R°; -OC(O)(CH2)Q^SR- SC(S)SR°; -(CH2)0^SC(O)RO; -(CH2)0-4C(O)NRO2; -C(S)NRO2; -C(S)SR°; -(CH2)O-4OC(0)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)RO; -C(NOR°)R°; -(CH2)O^SSR°; -(CH2)O^S(0)2R°; -(CH2)CMS(O)2OR°; -(CH2)0^OS(O)2RO; -S(O)2NR°2; -(CH2)O_4S(O)RO; -N(RO)S(O)2NR°2; -N(RO)S(O)2R°; -N(OR°)R°; -C(NH)NRO2; -P(O)2RO; -P(O)RO2; -OP(O)RO2; -OP(O)(ORO)2; SiR°3; -(C straight or branched alkylene)O-N(R°)2; or -(C-i^ straight or branched alkylene)C(O)O-N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C1-6 aliphatic, -CH2Ph, -0(CH2)o-iPh, -CH2-(5-6 membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0073] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, -(CH2)o--(CH2)O_2NHR’, -(CH2)O-2NR*2, -NO2, -SiR’3, -OSiR’3, -C(O)SR* -(Ci^ straight or branched alkylene)C(O)OR*, or-SSR* wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently selected from Ci_4aliphatic,-CH2Ph, -0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.

[0074] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted” group include the following: =0, =S, =NNR*2, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)2R*, =NR‘, =NOR‘, -O(C(R*2))2^O-, or -S(C(R*2))2-3S-, wherein each independent occurrence of R* is selected from hydrogen, Ci_6 aliphatic which may be substituted as defined below, or anunsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR*2)2-3O-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0075] Suitable substituents on the aliphatic group of R* include halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR’), -ON, -C(O)OH, -C(O)OR*, -NH2, -NHR*, -NR*2, or-N02, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C-i^ aliphatic, -CH2Ph, -0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0076] Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include -R+, -NR+2, -C(O)R+, -C(O)OR+, -C(O)C(O)R+, -C(O)CH2C(O)R+, -S(O)2R+, -S(O)2NR+2, -C(S)NR+2, -C(NH)NR+2, or -N(R+)S(O)2R+; wherein each R+ is independently hydrogen, C-i-e aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R+, taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0077] Suitable substituents on the aliphatic group of R+ are independently halogen, -R*, -(haloR*), -OH, -OR*, -0(haloR*), -CN, -C(O)OH, -C(O)OR*, -NH2, -NHR*, -NR*2, or-NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independentlyaliphatic, -CH2Ph, -0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0078] The term “leaving group” refers to an atom (or a group of atoms) with electron withdrawing ability that can be displaced as a stable species, taking with it the bonding electrons. Examples of suitable leaving groups include halides and sulfonate esters, including, but not limited to, triflate, mesylate, tosylate, and brosylate.

[0079] Compounds described herein can contain one or more double bonds and, thus, potentially give rise to cis / trans (E / Z) isomers, as well as other conformational isomers. Unless stated to the contrary, the invention includes all such possible isomers, as well as mixtures of such isomers.

[0080] Unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer and diastereomer, and a mixture of isomers, such as a racemic or scalemic mixture. Compounds described herein can contain one or more asymmetric centers and, thus, potentially give rise to diastereomers and optical isomers. Unless stated to the contrary, the present invention includes all such possible diastereomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof. Mixtures of stereoisomers, as well as isolated specific stereoisomers, are also included. During the course of the synthetic procedures used to prepare such compounds, or in using racemization or epimerization procedures known to those skilled in the art, the products of such procedures can be a mixture of stereoisomers.

[0081] Many organic compounds exist in optically active forms having the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes D and L or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or L meaning that the compound is levorotatory. A compound prefixed with (+) or D is dextrorotatory. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are non-superimposable mirror images of one another. A specific stereoisomer can also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture. Many of the compounds described herein can have one or more chiral centers and therefore can exist in different enantiomeric forms. If desired, a chiral carbon can be designated with an asterisk (*). When bonds to the chiral carbon are depicted as straight lines in the disclosed formulas, it is understood that both the (R) and (S) configurations of the chiral carbon, and hence both enantiomers and mixtures thereof, are embraced within the formula. As is used in the art, when it is desired to specify the absolute configuration about a chiral carbon, one of the bonds to the chiral carbon can be depicted as a wedge (bonds to atoms above the plane) and the other can be depicted as a series or wedge of short parallel lines is (bonds to atoms below the plane). The Cahn-lngold-Prelog system can be used to assign the (R) or (S) configuration to a chiral carbon.

[0082] Compounds described herein comprise atoms in both their natural isotopic abundance and in non-natural abundance. The disclosed compounds can be isotopically-labeled or isotopically-substituted compounds identical to those described, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine and chlorine, such as2H,3H,13C,14C,15N,180,170,35S,18F, and36CI, respectively. Compounds further comprise prodrugs thereof and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labeled compounds of the present invention, for example those into which radioactive isotopes such as3H and14C are incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, i.e.,3H, and carbon-14, i.e.,14C, isotopes are particularly preferred fortheir ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e.,2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labeled compounds of the present invention and prodrugs thereof can generally be prepared by carrying out the procedures below, by substituting a readily available isotopically labeled reagent for a non- isotopically labeled reagent.

[0083] The compounds described in the invention can be present as a solvate. In some cases, the solvent used to prepare the solvate is an aqueous solution, and the solvate is then often referred to as a hydrate. The compounds can be present as a hydrate, which can be obtained, for example, by crystallization from a solvent or from aqueous solution. In this connection, one, two, three or any arbitrary number of solvent or water molecules can combine with the compounds according to the invention to form solvates and hydrates. Unless stated to the contrary, the invention includes all such possible solvates.

[0084] It is also appreciated that certain compounds described herein can be present as an equilibrium of tautomers. For example, ketones with an a-hydrogen can exist in an equilibrium of the keto form and the enol form.keto form enol form amide form imidic acid form Likewise, amides with an / V-hydrogen can exist in an equilibrium of the amide form and theimidic acid form. Unless stated to the contrary, the invention includes all such possible tautomers.

[0085] It is known that chemical substances form solids which are present in different states of order which are termed polymorphic forms or modifications. The different modifications of a polymorphic substance can differ greatly in their physical properties. The compounds according to the invention can be present in different polymorphic forms, with it being possible for particular modifications to be metastable. Unless stated to the contrary, the invention includes all such possible polymorphic forms.

[0086] In some aspects, a structure of a compound can be represented by a formula:

[0087] which is understood to be equivalent to a formula:

[0088] wherein n is typically an integer. That is, Rnis understood to represent five independent substituents, Rn(a), Rn(b), Rn(c), Rn(d), and Rn(e). By “independent substituents,” it is meant that each R substituent can be independently defined. For example, if in one instance Rn(a)is halogen, then Rn(b)is not necessarily halogen in that instance.

[0089] As used herein, “administering” can refer to an administration that is oral, topical, intravenous, subcutaneous, transcutaneous, transdermal, intramuscular, intra-joint, parenteral, intra-arteriole, intradermal, intraventricular, intraosseous, intraocular, intracranial, intraperitoneal, intralesional, intranasal, intracardiac, intraarticular, intracavernous, intrathecal, intravireal, intracerebral, and intracerebroventricular, intratympanic, intracochlear, rectal, vaginal, by inhalation, by catheters, stents or via an implanted reservoir or other device that administers, either actively or passively (e.g. by diffusion) a composition the perivascular space and adventitia. For example, a medical device such as a stent can contain a composition or formulation disposed on its surface, which can then dissolve or be otherwise distributed to the surrounding tissue and cells. The term “parenteral” can include subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injections or infusion techniques. Administration can be continuous or intermittent. In various aspects, a preparation can be administered therapeutically; that is, administered to treat an existing disease or condition. Infurther various aspects, a preparation can be administered prophylactically; that is, administered for prevention of a disease or condition.

[0090] As used interchangeably herein, “subject,” “individual,” or “patient” can refer to a vertebrate organism, such as a mammal (e.g., human). "Subject" can also refer to a cell, a population of cells, a tissue, an organ, or an organism, preferably to human and constituents thereof.

[0091] As used herein, the terms "treating" and "treatment" can refer generally to obtaining a desired pharmacological and / or physiological effect. The effect can be, but does not necessarily have to be, prophylactic in terms of preventing or partially preventing a disease, symptom, or condition thereof, such as, for example, malaria or one or more symptoms associated therewith. The effect can be therapeutic in terms of a partial or complete cure of a disease, condition, symptom or adverse effect attributed to the disease, disorder, or condition. The term "treatment" as used herein can include any treatment of malaria or other parasitic infection in a subject, particularly a human and can include any one or more of the following: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., mitigating or ameliorating the disease and / or its symptoms or conditions. The term "treatment" as used herein can refer to both therapeutic treatment alone, prophylactic treatment alone, or both therapeutic and prophylactic treatment. Those in need of treatment (subjects in need thereof) can include those already with the disorder and / or those in which the disorder is to be prevented. As used herein, the term "treating", can include inhibiting the disease, disorder or condition, e.g., impeding its progress; and relieving the disease, disorder, or condition, e.g., causing regression of the disease, disorder and / or condition. Treating the disease, disorder, or condition can include ameliorating at least one symptom of the particular disease, disorder, or condition, even if the underlying pathophysiology is not affected, e.g., such as treating the pain of a subject by administration of an analgesic agent even though such agent does not treat the cause of the pain.

[0092] As used herein, the term “prevent” or “preventing” refers to precluding, averting, obviating, forestalling, stopping, or hindering something from happening, especially by advance action. It is understood that where reduce, inhibit, or prevent are used herein, unless specifically indicated otherwise, the use of the other two words is also expressly disclosed.

[0093] As used herein, “therapeutic” can refer to treating, healing, and / or ameliorating a disease, disorder, condition, or side effect, or to decreasing in the rate of advancement of a disease, disorder, condition, or side effect.

[0094] As used herein, “effective amount” can refer to the amount of a disclosed compound or pharmaceutical composition provided herein that is sufficient to effect beneficial or desired biological, emotional, medical, or clinical response of a cell, tissue, system, animal, or human. An effective amount can be administered in one or more administrations, applications, or dosages. The term can also include within its scope amounts effective to enhance or restore to substantially normal physiological function.

[0095] For example, it is well within the skill of the art to start doses of a compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, the effective daily dose can be divided into multiple doses for purposes of administration. Consequently, single dose compositions can contain such amounts or submultiples thereof to make up the daily dose. The dosage can be adjusted by the individual physician in the event of any contraindications. It is generally preferred that a maximum dose of the pharmacological agents of the invention (alone or in combination with other therapeutic agents) be used, that is, the highest safe dose according to sound medical judgment. It will be understood by those of ordinary skill in the art however, that a patient may insist upon a lower dose or tolerable dose for medical reasons, psychological reasons or for virtually any other reasons.

[0096] A response to a therapeutically effective dose of a disclosed compound and / or pharmaceutical composition, for example, can be measured by determining the physiological effects of the treatment or medication, such as the decrease or lack of disease symptoms following administration ofthe treatment or pharmacological agent. Other assays will be known to one of ordinary skill in the art and can be employed for measuring the level ofthe response. The amount of a treatment may be varied for example by increasing or decreasing the amount of a disclosed compound and / or pharmaceutical composition, by changing the disclosed compound and / or pharmaceutical composition administered, by changing the route of administration, by changing the dosage timing and so on. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products.

[0097] As used herein, the term “prophylactically effective amount” refers to an amount effective for preventing onset or initiation of a disease or condition.

[0098] The term “pharmaceutically acceptable” describes a material that is not biologically or otherwise undesirable, i.e., without causing an unacceptable level of undesirable biological effects or interacting in a deleterious manner.

[0099] The term “pharmaceutically acceptable salts”, as used herein, means salts of the active principal agents which are prepared with acids or bases that are tolerated by a biological system or tolerated by a subject or tolerated by a biological system and tolerated by a subject when administered in a therapeutically effective amount. When compounds of the present 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 solvent. Examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, ammonium, organic amino, magnesium salt, lithium salt, strontium salt or a similar salt. When compounds of the present 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, but are not limited to; 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.

[0100] The term “pharmaceutically acceptable prodrug” or “prodrug” represents those prodrugs of the compounds of the present disclosure which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for their intended use. Prodrugs of the present disclosure can be rapidly transformed in vivo to a parent compound having a structure of a disclosed compound, for example, by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, V. 14 of the A. C. S. Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).

[0101] As used herein, “dose,” “unit dose,” or “dosage” can refer to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of a disclosed compound and / or a pharmaceutical composition thereof calculated to produce the desired response or responses in association with its administration.

[0102] Certain materials, compounds, compositions, and components disclosed herein can be obtained commercially or readily synthesized using techniques generally known to those of skill in the art. For example, the starting materials and reagents used in preparing the disclosed compounds and compositions are either available from commercial suppliers such as Aldrich Chemical Co., (Milwaukee, Wis.), Acros Organics (Morris Plains, N. J.), Fisher Scientific (Pittsburgh, Pa.), or Sigma (St. Louis, Mo.) or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); March’s Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition); and Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., 1989).

[0103] Unless otherwise expressly stated, it is in no way intended that any method set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not actually recite an order to be followed by its steps or it is not otherwise specifically stated in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including: matters of logic with respect to arrangement of steps or operational flow; plain meaning derived from grammatical organization or punctuation; and the number or type of embodiments described in the specification.

[0104] Disclosed are the components to be used to prepare the compositions of the invention as well as the compositions themselves to be used within the methods disclosed herein. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds cannot be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed and a number of modifications that can be made to a number of molecules including the compounds are discussed, specifically contemplated is each and every combination and permutation of the compound and the modifications that are possible unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E, and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited each is individually and collectively contemplated meaning combinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are considered disclosed. Likewise, any subset or combinationof these is also disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the compositions of the invention. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the methods of the invention.

[0105] It is understood that the compositions disclosed herein have certain functions. Disclosed herein are certain structural requirements for performing the disclosed functions, and it is understood that there are a variety of structures that can perform the same function that are related to the disclosed structures, and that these structures will typically achieve the same result.

[0106] As used herein, the terms “optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0107] Unless otherwise specified, temperatures referred to herein are based on atmospheric pressure ( / .e. one atmosphere).Carboxamide Compounds and Methods for Making and Using Thereof

[0108] Described herein are compounds and methods for treating or preventing malaria in a subject. In one aspect, the compounds are imidazopyridine and purine carboxamide compounds. In one aspect, a compound having a formula represented by structure I or the pharmaceutically acceptable salt thereof can be used in the methods described hereinor a pharmaceutically acceptable salt thereof,wherein= represents a single or double bond;Z is CH or N;L1 is substituted or unsubstituted alkylene;L2 is substituted or unsubstituted alkylene group or substituted or unsubstituted cycloalkylene;R1 is H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocycloalkyl;R2 is H or alkyl, or R2 and L2 together with the N atom to which they are attached form a heterocycloalkyl;R3and R4are independently H, alkyl, or hydroxyalkyl, or R3and R4together with the N atom to which they are attached form a heterocycloalkyl; andY1, Y2, and Y3are independently selected from the group consisting of H, halide, and haloalkyl.

[0109] In one aspect, Z in structure I is CH. These compounds are referred to herein as imidazopyridine compounds. In another aspect, Z in structure I is N, which are referred to herein as purine compounds.

[0110] In one aspect, R1 in structure I is a substituted or unsubstituted phenyl. In one aspect, Li in structure I is an alkylene group. For example, Li is -(CH2)n-, where n is an integer from 1 to 6. In one aspect, Li is -(CH2)-. In another aspect, R1-L1 is Ph-(CH2)-, where Ph is substituted or unsubstituted phenyl.

[0111] In one aspect, R2 in structure I is H. In one aspect, L2 in structure I is an alkylene group. For example, L2 is -(CH2)m-, where m is an integer from 1 to 6. In one aspect, L2 in structure I is -(CH2)2-.

[0112] In one aspect, R3and R4in structure I are each H. In another aspect, R3is H and R4is alkyl such as, for example, a C1-C6 alkyl group (e.g., each a methyl group). In one aspect, R3and R4in structure I are each a C1-C6 alkyl group (e.g., each a methyl group).,,,, can be the same or different halide. In one aspect, Y1, Y2 and Y3 are independently Cl or F.In one aspect, Yi, Y2and Y3are two or three chloride groups. In another aspect,

[0114] In one aspect, the compound has the formula IIor the pharmaceutically acceptable salt thereof,whereinZ is CH or N;Ri is substituted or unsubstituted phenyl;R3and R4are independently H or alkyl; andYi, Y2, and Y3are independently selected from the group consisting of H and halide, wherein at least two of Y-i, Y2, and Y3are halide.

[0115] In one aspect, the compound has the formula III or IV

[0117] In one aspect, the compounds of structure I can be synthesized using the exemplary synthetic procedures provided in the Examples and FIGS. 1-3.Pharmaceutical Compositions

[0118] In various aspects, the present disclosure relates to pharmaceutical compositions comprising a therapeutically effective amount of at least one disclosed compound, at least oneproduct of a disclosed method, or a pharmaceutically acceptable salt thereof. As used herein,“pharmaceutically-acceptable carriers” means one or more of a pharmaceutically acceptable diluents, preservatives, antioxidants, solubilizers, emulsifiers, coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, and adjuvants. The disclosed pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy and pharmaceutical sciences.

[0119] In a further aspect, the disclosed pharmaceutical compositions comprise a therapeutically effective amount of at least one disclosed compound, at least one product of a disclosed method, or a pharmaceutically acceptable salt thereof as an active ingredient, a pharmaceutically acceptable carrier, optionally one or more other therapeutic agent, and optionally one or more adjuvant. The disclosed pharmaceutical compositions include those suitable for oral, rectal, topical, pulmonary, nasal, and parenteral administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. In a further aspect, the disclosed pharmaceutical composition can be formulated to allow administration orally, nasally, via inhalation, parenterally, paracancerally, transmucosally, transdermally, intramuscularly, intravenously, intradermally, subcutaneously, intraperitoneally, intraventricularly, intracranially and intratumorally.

[0120] As used herein, “parenteral administration” includes administration by bolus injection or infusion, as well as administration by intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular subarachnoid, intraspinal, epidural and intrasternal injection and infusion.

[0121] In various aspects, the present disclosure also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and, as active ingredient, a therapeutically effective amount of a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof. In a further aspect, a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof, or any subgroup or combination thereof may be formulated into various pharmaceutical forms for administration purposes.

[0122] In practice, the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, of the present disclosure can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceuticalcompounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present disclosure can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a nonaqueous liquid, as an oil-in-water emulsion or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compounds of the present disclosure, and / or pharmaceutically acceptable salt(s) thereof, can also be administered by controlled release means and / or delivery devices. The compositions can be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

[0123] It is especially advantageous to formulate the aforementioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. The term “unit dosage form,” as used herein, refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. That is, a “unit dosage form” is taken to mean a single dose wherein all active and inactive ingredients are combined in a suitable system, such that the patient or person administering the drug to the patient can open a single container or package with the entire dose contained therein and does not have to mix any components together from two or more containers or packages. Typical examples of unit dosage forms are tablets (including scored or coated tablets), capsules or pills for oral administration; single dose vials for injectable solutions or suspension; suppositories for rectal administration; powder packets; wafers; and segregated multiples thereof. This list of unit dosage forms is not intended to be limiting in any way, but merely to represent typical examples of unit dosage forms.

[0124] The pharmaceutical compositions disclosed herein comprise a compound of the present disclosure (or pharmaceutically acceptable salts thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally one or more additional therapeutic agents. In various aspects, the disclosed pharmaceutical compositions can include a pharmaceutically acceptable carrier and a disclosed compound, or a pharmaceutically acceptable salt thereof. In a further aspect, a disclosed compound, or pharmaceutically acceptable salt thereof, canalso be included in a pharmaceutical composition in combination with one or more other therapeutically active compounds. The instant compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

[0125] Techniques and compositions for making dosage forms useful for materials and methods described herein are described, for example, in the following references: Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G. Wilson, Eds.); Modern Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.).

[0126] The compounds described herein are typically to be administered in admixture with suitable pharmaceutical diluents, excipients, extenders, or carriers (termed herein as a pharmaceutically acceptable carrier, or a carrier) suitably selected with respect to the intended form of administration and as consistent with conventional pharmaceutical practices. The deliverable compound will be in a form suitable for oral, rectal, topical, intravenous injection or parenteral administration. Carriers include solids or liquids, and the type of carrier is chosen based on the type of administration being used. The compounds may be administered as a dosage that has a known quantity of the compound.

[0127] Because of the ease in administration, oral administration can be a preferred dosage form, and tablets and capsules represent the most advantageous oral dosage unit forms in which case solid pharmaceutical carriers are obviously employed. However, other dosage forms may be suitable depending upon clinical population (e.g., age and severity of clinical condition), solubility properties of the specific disclosed compound used, and the like.Accordingly, the disclosed compounds can be used in oral dosage forms such as pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. In preparing the compositions for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques.

[0128] The disclosed pharmaceutical compositions in an oral dosage form can comprise one or more pharmaceutical excipient and / or additive. Non-limiting examples of suitable excipients and additives include gelatin, natural sugars such as raw sugar or lactose, lecithin, pectin, starches (for example corn starch or amylose), dextran, polyvinyl pyrrolidone, polyvinyl acetate, gum arabic, alginic acid, tylose, talcum, lycopodium, silica gel (for example colloidal), cellulose, cellulose derivatives (for example cellulose ethers in which the cellulose hydroxy groups are partially etherified with lower saturated aliphatic alcohols and / or lower saturated, aliphatic oxyalcohols, for example methyl oxypropyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose phthalate), fatty acids as well as magnesium, calcium or aluminum salts of fatty acids with 12 to 22 carbon atoms, in particular saturated (for example stearates), emulsifiers, oils and fats, in particular vegetable (for example, peanut oil, castor oil, olive oil, sesame oil, cottonseed oil, corn oil, wheat germ oil, sunflower seed oil, cod liver oil, in each case also optionally hydrated); glycerol esters and polyglycerol esters of saturated fatty acids C12H24O2 to C-isHaeCh and their mixtures, it being possible for the glycerol hydroxy groups to be totally or also only partly esterified (for example mono-, di- and triglycerides); pharmaceutically acceptable mono- or multivalent alcohols and polyglycols such as polyethylene glycol and derivatives thereof, esters of aliphatic saturated or unsaturated fatty acids (2 to 22 carbon atoms, in particular 10-18 carbon atoms) with monovalent aliphatic alcohols (1 to 20 carbon atoms) or multivalent alcohols such as glycols, glycerol, diethylene glycol, pentacrythritol, sorbitol, mannitol and the like, which may optionally also be etherified, esters of citric acid with primary alcohols, acetic acid, urea, benzyl benzoate, dioxolanes, glyceroformals, tetrahydrofurfuryl alcohol, polyglycol ethers with C1-C12-alcohols, dimethylacetamide, lactamides, lactates, ethyl carbonates, silicones (in particular medium-viscous polydimethyl siloxanes), calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate, magnesium carbonate and the like.

[0129] Other auxiliary substances useful in preparing an oral dosage form are those which cause disintegration (so-called disintegrants), such as: cross-linked polyvinyl pyrrolidone, sodium carboxymethyl starch, sodium carboxymethyl cellulose or microcrystalline cellulose. Conventional coating substances may also be used to produce the oral dosage form. Those that may for example be considered are: polymerizates as well as copolymerizates of acrylic acid and / or methacrylic acid and / or their esters; copolymerizates of acrylic and methacrylic acid esters with a lower ammonium group content (for example EudragitR RS), copolymerizates of acrylic and methacrylic acid esters and trimethyl ammonium methacrylate (for example EudragitR RL); polyvinyl acetate; fats, oils, waxes, fatty alcohols; hydroxypropyl methyl cellulose phthalate or acetate succinate; cellulose acetate phthalate, starch acetate phthalate as well as polyvinyl acetate phthalate, carboxy methyl cellulose; methyl cellulose phthalate, methyl cellulose succinate, -phthalate succinate as well as methyl cellulose phthalic acid half ester; zein; ethyl cellulose as well as ethyl cellulose succinate; shellac, gluten; ethylcarboxyethyl cellulose; ethacrylate-maleic acid anhydride copolymer; maleic acid anhydride-vinyl methyl ether copolymer; styrol-maleic acid copolymerizate; 2-ethyl-hexyl-acrylate maleic acid anhydride; crotonic acid-vinyl acetate copolymer; glutaminic acid / glutamic acid ester copolymer; carboxymethylethylcellulose glycerol monooctanoate; cellulose acetate succinate; polyarginine.

[0130] Plasticizing agents that may be considered as coating substances in the disclosed oral dosage forms are: citric and tartaric acid esters (acetyl-triethyl citrate, acetyl tributyl-, tributyl-, triethyl-citrate); glycerol and glycerol esters (glycerol diacetate, -triacetate, acetylated monoglycerides, castor oil); phthalic acid esters (dibutyl-, diamyl-, diethyl-, dimethyl-, dipropylphthalate), di-(2-methoxy- or 2-ethoxyethyl)-phthalate, ethylphthalyl glycolate, butylphthalylethyl glycolate and butylglycolate; alcohols (propylene glycol, polyethylene glycol of various chain lengths), adipates (diethyladipate, di-(2-methoxy- or 2-ethoxyethyl)-adipate; benzophenone; diethyl- and diburylsebacate, dibutylsuccinate, dibutyltartrate; diethylene glycol dipropionate; ethyleneglycol diacetate, -dibutyrate, -dipropionate; tributyl phosphate, tributyrin; polyethylene glycol sorbitan monooleate (polysorbates such as Polysorbar 50); sorbitan monooleate.

[0131] Moreover, suitable binders, lubricants, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents may be included as carriers. The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include, but are not limited to, lactose, terra alba, sucrose, glucose, methylcellulose, dicalcium phosphate, calcium sulfate, mannitol, sorbitol talc, starch, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup,peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0132] In various aspects, a binder can include, for example, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. In a further aspect, a disintegrator can include, for example, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

[0133] In various aspects, an oral dosage form, such as a solid dosage form, can comprise a disclosed compound that is attached to polymers as targetable drug carriers or as a prodrug. Suitable biodegradable polymers useful in achieving controlled release of a drug include, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, caprolactones, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and hydrogels, preferably covalently crosslinked hydrogels.

[0134] Tablets may contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncoated, or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.

[0135] Atablet containing a disclosed compound can be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets can be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent.

[0136] In various aspects, a solid oral dosage form, such as a tablet, can be coated with an enteric coating to prevent ready decomposition in the stomach. In various aspects, enteric coating agents include, but are not limited to, hydroxypropylmethylcellulose phthalate, methacrylic acid-methacrylic acid ester copolymer, polyvinyl acetate-phthalate and cellulose acetate phthalate. Akihiko Hasegawa “Application of solid dispersions of Nifedipine withenteric coating agent to prepare a sustained-release dosage form” Chem. Pharm. Bull.33:1615-1619 (1985). Various enteric coating materials may be selected on the basis of testing to achieve an enteric coated dosage form designed ab initio to have a preferable combination of dissolution time, coating thicknesses and diametral crushing strength (e.g., see S. C. Porter et al. ‘‘The Properties of Enteric Tablet Coatings Made From Polyvinyl Acetatephthalate and Cellulose acetate Phthalate”, J. Pharm. Pharmacol. 22:42p (1970)). In a further aspect, the enteric coating may comprise hydroxypropyl-methylcellulose phthalate, methacrylic acid-methacrylic acid ester copolymer, polyvinyl acetate-phthalate and cellulose acetate phthalate.

[0137] In various aspects, an oral dosage form can be a solid dispersion with a water soluble or a water insoluble carrier. Examples of water soluble or water insoluble carrier include, but are not limited to, polyethylene glycol, polyvinylpyrrolidone, hydroxypropylmethyl-cellulose, phosphatidylcholine, polyoxyethylene hydrogenated castor oil, hydroxypropylmethylcellulose phthalate, carboxymethylethylcellulose, or hydroxypropylmethylcellulose, ethyl cellulose, or stearic acid.

[0138] In various aspects, an oral dosage form can be in a liquid dosage form, including those that are ingested, or alternatively, administered as a mouth wash or gargle. For example, a liquid dosage form can include aqueous suspensions, which contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. In addition, oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may also contain various excipients. The pharmaceutical compositions of the present disclosure may also be in the form of oil-in-water emulsions, which may also contain excipients such as sweetening and flavoring agents.

[0139] For the preparation of solutions or suspensions it is, for example, possible to use water, particularly sterile water, or physiologically acceptable organic solvents, such as alcohols (ethanol, propanol, isopropanol, 1,2-propylene glycol, polyglycols and their derivatives, fatty alcohols, partial esters of glycerol), oils (for example peanut oil, olive oil, sesame oil, almond oil, sunflower oil, soya bean oil, castor oil, bovine hoof oil), paraffins, dimethyl sulfoxide, triglycerides and the like.

[0140] In the case of a liquid dosage form such as a drinkable solutions, the following substances may be used as stabilizers or solubilizers: lower aliphatic mono- and multivalent alcohols with 2-4 carbon atoms, such as ethanol, n-propanol, glycerol, polyethylene glycols with molecular weights between 200-600 (for example 1 to 40% aqueous solution), diethylene glycol monoethyl ether, 1,2-propylene glycol, organic amides, for example amides of aliphaticC1-C6-carboxylic acids with ammonia or primary, secondary or tertiary C1-C4-amines or C1-C4-hydroxy amines such as urea, urethane, acetamide, N-methyl acetamide, N, N-diethyl acetamide, N, N-dimethyl acetamide, lower aliphatic amines and diamines with 2-6 carbon atoms, such as ethylene diamine, hydroxyethyl theophylline, tromethamine (for example as 0.1 to 20% aqueous solution), aliphatic amino acids.

[0141] In preparing the disclosed liquid dosage form can comprise solubilizers and emulsifiers such as the following non-limiting examples can be used: polyvinyl pyrrolidone, sorbitan fatty acid esters such as sorbitan trioleate, phosphatides such as lecithin, acacia, tragacanth, polyoxyethylated sorbitan monooleate and other ethoxylated fatty acid esters of sorbitan, polyoxyethylated fats, polyoxyethylated oleotriglycerides, linolizated oleotriglycerides, polyethylene oxide condensation products of fatty alcohols, alkylphenols or fatty acids or also 1-methyl-3-(2-hydroxyethyl)imidazolidone-(2). In this context, polyoxyethylated means that the substances in question contain polyoxyethylene chains, the degree of polymerization of which generally lies between 2 and 40 and in particular between 10 and 20. Polyoxyethylated substances of this kind may for example be obtained by reaction of hydroxyl group-containing compounds (for example mono- or diglycerides or unsaturated compounds such as those containing oleic acid radicals) with ethylene oxide (for example 40 Mol ethylene oxide per 1 Mol glyceride). Examples of oleotriglycerides are olive oil, peanut oil, castor oil, sesame oil, cottonseed oil, corn oil. See also Dr. H. P. Fiedler “Lexikon der Hillsstoffe fur Pharmazie, Kostnetik und angrenzende Gebiete” 1971, pages 191-195.

[0142] In various aspects, a liquid dosage form can further comprise preservatives, stabilizers, buffer substances, flavor correcting agents, sweeteners, colorants, antioxidants and complex formers and the like. Complex formers which may be for example be considered are: chelate formers such as ethylene diamine retrascetic acid, nitrilotriacetic acid, diethylene triamine pentacetic acid and their salts.

[0143] It may optionally be necessary to stabilize a liquid dosage form with physiologically acceptable bases or buffers to a pH range of approximately 6 to 9. Preference may be given to as neutral or weakly basic a pH value as possible (up to pH 8).

[0144] In order to enhance the solubility and / or the stability of a disclosed compound in a disclosed liquid dosage form, a parenteral injection form, or an intravenous injectable form, it can be advantageous to employ a-, p- or y-cyclodextrins or their derivatives, in particular hydroxyalkyl substituted cyclodextrins, e.g. 2-hydroxypropyl-p-cyclodextrin or sulfobutyl-p-cyclodextrin. Also co-solvents such as alcohols may improve the solubility and / or the stability of the compounds according to the present disclosure in pharmaceutical compositions.

[0145] In various aspects, a disclosed liquid dosage form, a parenteral injection form, or an intravenous injectable form can further comprise liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.

[0146] Pharmaceutical compositions of the present disclosure suitable injection, such as parenteral administration, such as intravenous, intramuscular, or subcutaneous administration. Pharmaceutical compositions for injection can be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.

[0147] Pharmaceutical compositions of the present disclosure suitable for parenteral administration can include sterile aqueous or oleaginous solutions, suspensions, or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In some aspects, the final injectable form is sterile and must be effectively fluid for use in a syringe. The pharmaceutical compositions should be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

[0148] Injectable solutions, for example, can be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. In some aspects, a disclosed parenteral formulation can comprise about 0.01-0.1 M, e.g. about 0.05 M, phosphate buffer. In a further aspect, a disclosed parenteral formulation can comprise about 0.9% saline.

[0149] In various aspects, a disclosed parenteral pharmaceutical composition can comprise pharmaceutically acceptable carriers such as aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include but not limited to water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles can include mannitol, normal serum albumin, sodium chloride solution, Ringer's dextrose, dextroseand sodium chloride, lactated Ringer's and fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers such as those based on Ringer's dextrose, and the like. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, collating agents, inert gases and the like. In a further aspect, a disclosed parenteral pharmaceutical composition can comprise may contain minor amounts of additives such as substances that enhance isotonicity and chemical stability, e.g., buffers and preservatives. Also contemplated for injectable pharmaceutical compositions are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations. Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the subject or patient.

[0150] In addition to the pharmaceutical compositions described herein above, the disclosed compounds can also be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds 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.

[0151] Pharmaceutical compositions of the present disclosure can be in a form suitable for topical administration. As used herein, the phrase “topical application” means administration onto a biological surface, whereby the biological surface includes, for example, a skin area (e.g., hands, forearms, elbows, legs, face, nails, anus and genital areas) or a mucosal membrane. By selecting the appropriate carrier and optionally other ingredients that can be included in the composition, as is detailed herein below, the compositions of the present invention may be formulated into any form typically employed for topical application. Atopical pharmaceutical composition can be in a form of a cream, an ointment, a paste, a gel, a lotion, milk, a suspension, an aerosol, a spray, foam, a dusting powder, a pad, and a patch. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations can be prepared, utilizing a compound of the present disclosure, or pharmaceutically acceptable salts thereof, via conventional processing methods. As an example, a cream or ointment is prepared by mixing hydrophilic material and water, together with about 5 wt% to about 10wt% of the compound, to produce a cream or ointment having a desired consistency.

[0152] In the compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and / or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not introduce a significant deleterious effect on the skin. Said additives may facilitate the administration to the skin and / or may be helpful for preparing the desired compositions. These compositions maybe administered in various ways, e.g., as a transdermal patch, as a spot-on, as an ointment.

[0153] Ointments are semisolid preparations, typically based on petrolatum or petroleum derivatives. The specific ointment base to be used is one that provides for optimum delivery for the active agent chosen for a given formulation, and, preferably, provides for other desired characteristics as well (e.g., emollience). As with other carriers or vehicles, an ointment base should be inert, stable, nonirritating and nonsensitizing. As explained in Remington: The Science and Practice of Pharmacy, 19th Ed., Easton, Pa.: Mack Publishing Co. (1995), pp.1399-1404, ointment bases may be grouped in four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases include, for example, vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, and include, for example, cetyl alcohol, glyceryl monostearate, lanolin and stearic acid. Preferred water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weight.

[0154] Lotions are preparations that are to be applied to the skin surface without friction. Lotions are typically liquid or semiliquid preparations in which solid particles, including the active agent, are present in a water or alcohol base. Lotions are typically preferred for treating large body areas, due to the ease of applying a more fluid composition. Lotions are typically suspensions of solids, and oftentimes comprise a liquid oily emulsion of the oil-in-water type. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the active agent in contact with the skin, such as methylcellulose, sodium carboxymethyl-cellulose, and the like.

[0155] Creams are viscous liquids or semisolid emulsions, either oil-in-water or water-in-oil. Cream bases are typically water-washable, and contain an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the “internal” phase, is generally comprised of petrolatum and / or a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase typically, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant. The emulsifier in a cream formulation is generally a nonionic, anionic, cationic or amphoteric surfactant. Reference may be made to Remington: The Science and Practice of Pharmacy, supra, for further information.

[0156] Pastes are semisolid dosage forms in which the bioactive agent is suspended in a suitable base. Depending on the nature of the base, pastes are divided between fatty pastesor those made from a single-phase aqueous gel. The base in a fatty paste is generally petrolatum, hydrophilic petrolatum and the like. The pastes made from single-phase aqueous gels generally incorporate carboxymethylcellulose or the like as a base. Additional reference may be made to Remington: The Science and Practice of Pharmacy, for further information.

[0157] Gel formulations are semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the carrier liquid, which is typically aqueous, but also, preferably, contain an alcohol and, optionally, an oil. Preferred organic macromolecules, i.e., gelling agents, are crosslinked acrylic acid polymers such as the family of carbomer polymers, e.g., carboxypolyalkylenes that may be obtained commercially under the trademark Carbopol™. Other types of preferred polymers in this context are hydrophilic polymers such as polyethylene oxides, polyoxyethylenepolyoxypropylene copolymers and polyvinylalcohol; modified cellulose, such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. In order to prepare a uniform gel, dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing or stirring, or combinations thereof.

[0158] Sprays generally provide the active agent in an aqueous and / or alcoholic solution which can be misted onto the skin for delivery. Such sprays include those formulated to provide for concentration of the active agent solution at the site of administration following delivery, e.g., the spray solution can be primarily composed of alcohol or other like volatile liquid in which the active agent can be dissolved. Upon delivery to the skin, the carrier evaporates, leaving concentrated active agent at the site of administration.

[0159] Foam compositions are typically formulated in a single or multiple phase liquid form and housed in a suitable container, optionally together with a propellant which facilitates the expulsion of the composition from the container, thus transforming it into a foam upon application. Other foam forming techniques include, for example the “Bag-in-a-can” formulation technique. Compositions thus formulated typically contain a low-boiling hydrocarbon, e.g., isopropane. Application and agitation of such a composition at the body temperature cause the isopropane to vaporize and generate the foam, in a manner similar to a pressurized aerosol foaming system. Foams can be water-based or aqueous alkanolic, but are typically formulated with high alcohol content which, upon application to the skin of a user, quickly evaporates, driving the active ingredient through the upper skin layers to the site of treatment.

[0160] Skin patches typically comprise a backing, to which a reservoir containing the active- M -agent is attached. The reservoir can be, for example, a pad in which the active agent or composition is dispersed or soaked, or a liquid reservoir. Patches typically further include a frontal water permeable adhesive, which adheres and secures the device to the treated region. Silicone rubbers with self-adhesiveness can alternatively be used. In both cases, a protective permeable layer can be used to protect the adhesive side of the patch prior to its use. Skin patches may further comprise a removable cover, which serves for protecting it upon storage.

[0161] Examples of patch configuration which can be utilized with the present invention include a single-layer or multi-layer drug-in-adhesive systems which are characterized by the inclusion of the drug directly within the skin-contacting adhesive. In such a transdermal patch design, the adhesive not only serves to affix the patch to the skin, but also serves as the formulation foundation, containing the drug and all the excipients under a single backing film. In the multi-layer drug-in-adhesive patch a membrane is disposed between two distinct drug-in-adhesive layers or multiple drug-in-adhesive layers are incorporated under a single backing film.

[0162] Examples of pharmaceutically acceptable carriers that are suitable for pharmaceutical compositions for topical applications include carrier materials that are well-known for use in the cosmetic and medical arts as bases for e.g., emulsions, creams, aqueous solutions, oils, ointments, pastes, gels, lotions, milks, foams, suspensions, aerosols and the like, depending on the final form of the composition. Representative examples of suitable carriers according to the present invention therefore include, without limitation, water, liquid alcohols, liquid glycols, liquid polyalkylene glycols, liquid esters, liquid amides, liquid protein hydrolysates, liquid alkylated protein hydrolysates, liquid lanolin and lanolin derivatives, and like materials commonly employed in cosmetic and medicinal compositions. Other suitable carriers according to the present invention include, without limitation, alcohols, such as, for example, monohydric and polyhydric alcohols, e.g., ethanol, isopropanol, glycerol, sorbitol, 2-methoxyethanol, diethylene glycol, ethylene glycol, hexyleneglycol, mannitol, and propylene glycol; ethers such as diethyl or dipropyl ether; polyethylene glycols and methoxypolyoxyethylenes (carbowaxes having molecular weight ranging from 200 to 20,000); polyoxyethylene glycerols, polyoxyethylene sorbitols, stearoyl diacetin, and the like.

[0163] Topical compositions of the present disclosure can, if desired, be presented in a pack or dispenser device, such as an FDA-approved kit, which may contain one or more unit dosage forms containing the active ingredient. The dispenser device may, for example, comprise a tube. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser device may also be accompanied by a notice in a form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, whichnotice is reflective of approval by the agency of the form of the compositions for human or veterinary administration. Such notice, for example, may include labeling approved by the U. S. Food and Drug Administration for prescription drugs or of an approved product insert. Compositions comprising the topical composition of the invention formulated in a pharmaceutically acceptable carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0164] Another patch system configuration which can be used by the present invention is a reservoir transdermal system design which is characterized by the inclusion of a liquid compartment containing a drug solution or suspension separated from the release liner by a semi-permeable membrane and adhesive. The adhesive component of this patch system can either be incorporated as a continuous layer between the membrane and the release liner or in a concentric configuration around the membrane. Yet another patch system configuration which can be utilized by the present invention is a matrix system design which is characterized by the inclusion of a semisolid matrix containing a drug solution or suspension which is in direct contact with the release liner. The component responsible for skin adhesion is incorporated in an overlay and forms a concentric configuration around the semisolid matrix.

[0165] Pharmaceutical compositions of the present disclosure can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butterand other materials commonly used in the art. The suppositories can be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

[0166] Pharmaceutical compositions containing a compound of the present disclosure, and / or pharmaceutically acceptable salts thereof, can also be prepared in powder or liquid concentrate form.

[0167] The pharmaceutical composition (or formulation) may be packaged in a variety of ways. Generally, an article for distribution includes a container that contains the pharmaceutical composition in an appropriate form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, foil blister packs, and the like. The container may also include a tamper proof assemblage to prevent indiscreet access to the contents of the package. In addition, the container typically has deposited thereon a label that describes the contents of the container and any appropriate warnings or instructions.

[0168] The disclosed pharmaceutical compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the activeingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U. S. Food and Drug Administration for prescription drugs, or the approved product insert. Pharmaceutical compositions comprising a disclosed compound formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0169] The exact dosage and frequency of administration depends on the particular disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, solvate, or polymorph thereof, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof; the particular condition being treated and the severity of the condition being treated; various factors specific to the medical history of the subject to whom the dosage is administered such as the age; weight, sex, extent of disorder and general physical condition of the particular subject, as well as other medication the individual may be taking; as is well known to those skilled in the art. Furthermore, it is evident that said effective daily amount may be lowered or increased depending on the response of the treated subject and / or depending on the evaluation of the physician prescribing the compounds of the present disclosure.

[0170] Depending on the mode of administration, the pharmaceutical composition will comprise from 0.05 to 99% by weight, preferably from 0.1 to 70% by weight, more preferably from 0.1 to 50% by weight of the active ingredient, and, from 1 to 99.95% by weight, preferably from 30 to 99.9% by weight, more preferably from 50 to 99.9% by weight of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.

[0171] In one aspect, an appropriate dosage level will generally be about 0.01 to 1000 mg of a compound described herein per kg patient body weight per day and can be administered in single or multiple doses. In various aspects, the dosage level will be about 0.1 to about 500 mg / kg per day, about 0.1 to 250 mg / kg per day, or about 0.5 to 100 mg / kg per day. A suitable dosage level can be about 0.01 to 1000 mg / kg per day, about 0.01 to 500 mg / kg per day, about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range the dosage can be 0.05 to 0.5, 0.5 to 5.0 or 5.0 to 50 mg / kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 mg of the active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75,100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000 mg of the active ingredient for the symptomatic adjustment of the dosage of the patient to be treated. The compound can be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosing regimen can be adjusted to provide the optimal therapeutic response.

[0172] Such unit doses as described hereinabove and hereinafter can be administered more than once a day, for example, 2, 3, 4, 5 or 6 times a day. In various aspects, such unit doses can be administered 1 or 2 times per day, so that the total dosage for a 70 kg adult is in the range of 0.001 to about 15 mg per kg weight of subject per administration. In a further aspect, dosage is 0.01 to about 1.5 mg per kg weight of subject per administration, and such therapy can extend for a number of weeks or months, and in some cases, years. It will be understood, however, that the specific dose level for any particular patient will depend on a variety of factors including the activity of the specific compound employed; the age, body weight, general health, sex and diet of the individual being treated; the time and route of administration; the rate of excretion; other drugs that have previously been administered; and the severity of the particular disease undergoing therapy, as is well understood by those of skill in the area.

[0173] A typical dosage can be one 1 mg to about 100 mg tablet or 1 mg to about 300 mg taken once a day, or multiple times per day, or one time-release capsule or tablet taken once a day and containing a proportionally higher content of active ingredient. The time-release effect can be obtained by capsule materials that dissolve at different pH values, by capsules that release slowly by osmotic pressure, or by any other known means of controlled release.

[0174] It can be necessary to use dosages outside these ranges in some cases as will be apparent to those skilled in the art. Further, it is noted that the clinician or treating physician will know how and when to start, interrupt, adjust, or terminate therapy in conjunction with individual patient response.

[0175] The disclosed pharmaceutical compositions can further comprise other therapeutically active compounds, which are usually applied in the treatment of the above mentioned pathological or clinical conditions.

[0176] It is understood that the disclosed compositions can be prepared from the disclosed compounds. It is also understood that the disclosed compositions can be employed in the disclosed methods of using.

[0177] As already mentioned, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, and a pharmaceutically acceptable carrier. Additionally, thepresent disclosure relates to a process for preparing such a pharmaceutical composition, characterized in that a pharmaceutically acceptable carrier is intimately mixed with a therapeutically effective amount of a compound according to the present disclosure.Methods for Treatment and Prevention of Malaria and Other Parasitic Infections

[0178] Human malaria is caused by single-celled microorganisms of the Plasmodium genus. It is spread exclusively through bites of infected female Anopheles mosquitoes. The mosquito bite introduces the parasites, and the parasites travel to the liver, where they mature and reproduce. Host infection initiates with sporozoite invasion of hepatocytes, followed by a dramatic parasite amplification event during liver stage (also referred to as exoerythrocytic form (EEF)). Following liver stage development, the parasites enter the blood stream (i.e., the blood stage), where they invade red blood cells (erythrocytes). Expansion of the blood stage results in the clinical manifestation of malaria, where symptoms such as fever, chills, headaches, muscle and joint pain, abdominal pain, nausea and vomiting, and diarrhea can occur.

[0179] In one aspect, disclosed herein is a method for the treatment or prevention of a malaria in a subject, the method including the step of administering to the subject a therapeutically effective amount of at least one disclosed compound, or a pharmaceutically acceptable salt thereof, or the disclosed pharmaceutical composition. In some aspects, the subject is a human. In another aspect, the subject has been diagnosed with a need for treatment of malaria prior to the administering step. In some aspects, the method further includes the step of identifying a subject in need of treatment of malaria.

[0180] In one aspect, the compounds described herein exhibit potent antimalarial activity against one or more species of the genus Plasmodium. In one aspect, the malaria parasite comprises Plasmodium falciparum, Plasmodium vivax, Plasmodium knowlesi, Plasmodium ovalecurtisi, Plasmodium ovalewallikeri, or Plasmodium malariae. In another aspect, the compounds described herein inhibit or prevent the blood stage of malaria. In one aspect, the compounds described herein can eliminate the parasite present in the blood of an infected subject after 30 days of administration of the compound to the subject.

[0181] The compounds described herein are effective in treating one or more symptoms of malaria. For example, the compounds described herein can treat swollen lymph nodes, fever, moderate-to-severe shaking chills, profuse sweating, diarrhea, anemia, cough, muscle and join aches, fatigue, headache, rapid breathing, rapid heart rate, sore throat, nausea and vomiting, enlarged spleen, mild jaundice, enlargement of the liver, brain inflammation and swelling (cerebral malaria), seizures, confusion or impaired consciousness, vision problems,coma, circulatory collapse / shock, pulmonary edema or acute respiratory distress syndrome (ARDS), acidosis, acute kidney injury, abnormal bleeding or disseminated intravascular coagulation (DIC).

[0182] In addition to treating or preventing malaria, the compounds are effective in treating or preventing infections caused by other parasites. In one aspect, the compounds described herein can be used to treat or prevent an infection caused by Trypanosoma cruzi. Trypanosoma cruzi is a species of parasitic euglenoids which causes Chagas disease. Trypanosomiasis in humans progresses with the development of the trypanosome into a trypomastigote in the blood and into an amastigote in tissues. As the infection progresses, the number of infected cells increases, as well as the number of amastigotes per infected cell (APC). In one aspect, the compounds described herein can remove the parasite from the blood and infected tissues (organs) after administration to the subject. In another aspect, the compounds described herein can treat one or more symptoms due to Trypanosoma cruzi infection.

[0183] In one aspect, the compounds described herein can be used to treat or prevent an infection caused by Naegleria fowle. Naegleria fowled, also known as the brain-eating amoeba, is a species of the genus Naegleria. In one aspect, the compounds described herein can remove the parasite from the brain after administration to the subject. In one aspect, the compounds described herein can protect the brain tissue (cytoprotection). In another aspect, the compounds described herein can treat one or more symptoms due to Naegleria fowler! infection.

[0184] In one aspect, the compound is administered orally to the subject. In another aspect, the compound is administered intravenously or intramuscularly to the subject. In one aspect, the compound is administered at a dosage of from about 50 mg per day to about 1,000 mg per day, or about 50 mg per day, 50 mg per day, 100 mg per day, 150 mg per day, 200 mg per day, 250 mg per day, 300 mg per day, 350 mg per day, 400 mg per day, 450 mg per day, 500 mg per day, 550 mg per day, 600 mg per day, 650 mg per day, 700 mg per day, 750 mg per day, 800 mg per day, 850 mg per day, 900 mg per day, 950 mg per day, or 1,000 mg per day, where any value can be a lower and upper endpoint of a range (e.g., 100 mg per day to 300 mg per day).

[0185] Now having described the aspects of the present disclosure, in general, the following Examples describe some additional aspects of the present disclosure. While aspects of the present disclosure are described in connection with the following examples and the corresponding text and figures, there is no intent to limit aspects of the present disclosure to this description. On the contrary, the intent is to cover all alternatives, modifications, andequivalents included within the spirit and scope of the present disclosure.Aspects

[0186] Aspect 1. A compound of formula Ior a pharmaceutically acceptable salt thereof,wherein— represents a single or double bond;Z is CH or N;L1 is substituted or unsubstituted alkylene;l_2is substituted or unsubstituted alkylene group or substituted or unsubstituted cycloalkylene;Ri is H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocycloalkyl;R2is H or alkyl, or R2and l_2together with the N atom to which they are attached form a heterocycloalkyl;R3and R4are independently H, alkyl, or hydroxyalkyl, or R3and R4together with the N atom to which they are attached form a heterocycloalkyl; andY1, Y2, and Y3are independently selected from the group consisting of H, halide, and haloalkyl.

[0187] Aspect 2. The compound of Aspect 1, wherein Z is CH.

[0188] Aspect 3. The compound of Aspect 1, wherein Z is N.

[0189] Aspect 4. The compound of any one of Aspects 1-3, wherein Ri is substituted or unsubstituted phenyl.

[0190] Aspect 5. The compound of any one of Aspects 1 -4, wherein Li is -(CH2)-.

[0191] Aspect 6. The compound of any one of Aspects 1-5, wherein R2is H.

[0192] Aspect 7. The compound of any one of Aspects 1-6, wherein L2is -(CH2)2-.

[0193] Aspect 8. The compound of any one of Aspects 1-7, wherein R3and R4are each H.

[0194] Aspect 9. The compound of any one of Aspects 1-7, wherein R3is H and R4is alkyl.

[0195] Aspect 10. The compound of any one of Aspects 1-9, wherein

[0196] Aspect 11. The compound of any one of Aspects 1-10, wherein Y1, Y2and Y3are independently Cl or F.

[0197] Aspect 12. The compound of any one of Aspects 1-11, whereinis agroup of formula

[0198] Aspect 13. The compound of Aspect 1, wherein the compound has the formula IIor the pharmaceutically acceptable salt thereof,whereinZ is CH or N;Ri is substituted or unsubstituted phenyl;R3and R4are independently H or alkyl; andYi, Y2, and Y3are independently selected from the group consisting of H and halide, wherein at least two of Yi, Y2, and Y3are halide.

[0199] Aspect 14. The compound of Aspect 13, wherein Z is CH.

[0200] Aspect 15. The compound of Aspect 13, wherein Z is N.

[0201] Aspect 16. The compound of any one of Aspects 13-15, wherein R1 is unsubstituted phenyl.

[0202] Aspect 17. The compound of any one of Aspects 13-16, wherein R3 and R4 are each H.

[0203] Aspect 18. The compound of any one of Aspects 13-17, wherein Y1, Y2, and Y3 are each chloro.w Iw

[0204] Aspect 19. The compound of any one of Aspects 13-17, whereinjSagroup of formula

[0205] Aspect 20. The compound of Aspect 1, wherein the compound has the formula III orIII IVor the pharmaceutically acceptable salt thereof.

[0206] Aspect 21. A pharmaceutical composition comprising the compound of any one of Aspects 1-20 and a pharmaceutically acceptable carrier.

[0207] Aspect 22. A method for treating or preventing malaria in a subject, the method comprising administering to the subject an effective amount of the compound of any one of Aspects 1-20.

[0208] Aspect 23. The method of Aspect 22, wherein the subject is infected with a strain of Plasmodium spp.

[0209] Aspect 24. The method of Aspect 23, wherein the strain of Plasmodium spp. comprises Plasmodium falciparum, Plasmodium vivax, Plasmodium knowlesi, Plasmodium ovalecurtisi, Plasmodium ovalewallikeri, or Plasmodium malariaei.

[0210] Aspect 25. The method of any one of Aspects 22-24, wherein the method treats one or more symptoms of malaria.

[0211] Aspect 26. The method of Aspect 25, wherein the symptom comprises swollen lymph nodes, fever, moderate-to-severe shaking chills, profuse sweating, diarrhea, anemia, cough, muscle and join aches, fatigue, headache, rapid breathing, rapid heart rate, sore throat, nausea and vomiting, enlarged spleen, mild jaundice, enlargement of the liver, brain inflammation and swelling (cerebral malaria), seizures, confusion or impaired consciousness, vision problems, coma, circulatory collapse / shock, pulmonary edema or acute respiratory distress syndrome (ARDS), acidosis, acute kidney injury, abnormal bleeding or disseminated intravascular coagulation (DIC).

[0212] Aspect 27. The method of any one of Aspects 22-26, wherein after administration of the compound, a parasite that infected the subject with malaria is no longer present in the blood of the subject after 30 days of administration of the compound to the subject.

[0213] Aspect 28. A method for treating or preventing an infection by Trypanosoma cruzi or Naegleria fowled in a subject, the method comprising administering to the subject an effective amount of the compound of any one of Aspects 1-20.

[0214] Aspect 29. The method of any one of Aspects 22-28, wherein the compound is administered orally, intravenously, or intramuscularly to the subject.

[0215] Aspect 30. A compound of formula XIXIor a pharmaceutically acceptable salt thereof,wherein= represents a single or double bond;Z is CH or N;Li is alkylene optionally substituted with hyrdroxy;I_2is alkylene or cycloalkylene optionally substituted with hydroxy;Ri is H, alkyl, aryl, cycloalkyl, heteroaryl, or heterocyclyl, wherein the alkyl, aryl, cycloalkyl, heteroaryl, or heterocyclyl is optionally substituted with one or more alkyl, halo, haloalkyl, cyano, pentafluorosulfanyl, or a combination thereof; R2is H or alkyl, or R2and l_2together with the N atom to which they are attached form a heterocyclyl;R2is alkylene or cycloalkylene optionally substituted with hydroxy;R3and R4are independently H, alkyl, or hydroxyalkyl, or R3and R4together with the N atom to which they are attached form a heterocyclyl; andYi, Y2, and Y3are independently selected from H, halo, haloalkyl.

[0216] Aspect 31. A compound according to Aspect 30 where Z is N.

[0217] Aspect 32. A compound according to Aspect 30 wherein Z is CH.

[0218] Aspect 33. A compound according to Aspect 30 wherein

[0219] Aspect 34. A compound according to Aspect 33 wherein Y1, Y2and Y3are independently Cl or F.

[0220] Aspect 35. A compound according to Aspect 34 wherein

[0221] Aspect 36. A compound according to Aspect 30 wherein Ri is alkyl.

[0222] Aspect 37. A compound according to Aspect 30 wherein Ri is phenyl.

[0223] Aspect 38. A compound according to Aspect 30 wherein Li is -CH2-.

[0224] Aspect 39. A compound according to Aspect 30 wherein l_2is alkylene optionally substituted with hydroxy.

[0225] Aspect 40. A compound according to Aspect 30 wherein R3and R4are independently H or alkyl.

[0226] Aspect 41. A composition comprising a compound according to Aspect 30 and a pharmaceutically acceptable diluent or excipient.

[0227] Aspect 42. A method of treating malaria in a subject comprising administering to the subject an effective amount of a compound according to Aspect 30.EXAMPLES

[0228] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure.

[0229] MATERIALS AND METHODS

[0230] Synthesis of Imidazopyridine Analogs

[0231] General

[0232] All chemicals were purchased from a number of vendors. Imidazo[4,5-c]pyridine methyl esters 6d-15d, and amides 6a-14a, 6b-9b, 11b-14b, 6c-8c, 11c-15c were prepared by using described method below. Exemplary reaction schemes for producing the imidazopyridine analogs are provided in FIGS. 1 and 2.

[0233] Compounds were purchased from various vendor and were used without purification, unless otherwise noted.1H NMR spectra were recorded at 400, 500 or 600 MHz; the corresponding13C NMR resonant frequencies were 101, 126 and 151 MHz respectively; the corresponding19F NMR resonant frequencies were 376 MHz and these spectra were1H-decoupled.

[0234] General Procedure A: Pictet -Spengler, esterification, oxidation, and benzylation procedures for aromatic aldehydes

[0235] (i) Pictet-Spengler

[0236] To a 50 mL flask containing histidine (1.0 g), potassium hydroxide (2.0 equiv) and the requisite aldehyde (0.98 - 1.01 equiv), ethanol-water (1:1, 15 mL) was added. A reflux condenser was added, and the mixture was stirred at reflux for 1 h. After cooling to room temperature, aq HCI (1 M) was added dropwise until neutrality (pH paper). Concentration in vacuo, dissolution in methanol, filtration, and concentration in vacuo was performed to afford the crude diastereomeric Pictet-Spengler adducts, typically as a light-yellow solid. This material was used in the next step without further purification.

[0237] (ii) Esterification

[0238] A 50 ml_ flask was charged with the crude Pictet-Spengler material described above (1.0 g) and methanol (20 ml_) and cooled to 0°. Thionyl chloride (2 equiv) was added dropwise in methanol, a reflux condenser was added, and the mixture was heated at reflux for 16 h. The mixture was cooled to room temperature, and saturated aq. sodium bicarbonate solution (50 mL) was added dropwise until the neutrality (pH paper). Concentration in vacuo, dissolution in methanol, filtration, and concentration in vacuo was performed to afford the crude diastereomeric Pictet-Spengler methyl esters, typically as a light-yellow solid. The crude material was used in the next step without further purification.

[0239] (Hi) Oxidation

[0240] To a 50 mL flask containing the crude diastereomeric Pictet-Spengler methyl esters (1.0 g), Dess-Martin Periodinane (DMP, 2.2 equiv) was added in small portions in dichloromethane (X mL) at 0°C. The flask was capped and stirred at room temperature for 24 h. TLC monitoring of reaction progress is convenient; the imidazo[4,5-c]pyridine products typically appear above the starting materials. Once the starting material is fully consumed, the reaction mixture was poured into saturated aq. sodium bicarbonate solution (20 mL); after stirring, the precipitate was collected on a filter paper then washed with dichloromethane (3 x 15 mL) to obtain a white solid. In most cases, the precipitate was dried and used without further purification. But full characterization of 3a is provided below.

[0241] General Procedure B: Benzylation of imidazo[4,5-c]pyridine methyl ester

[0242] To a 20 mL vial containing the appropriate oxidation crude material (200mg)and cesium carbonate (1.5 equiv), benzyl bromide (1.2 equiv) was added dropwise in DMF at room temperature. The vial was capped and stirring at 80°C for 16h, the reaction was cooled to room temperature. TLC typically indicated complete consumption of the oxidation intermediate. The reaction mixture was extracted with EtOAc (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The benzylation product was further purified by silica gel column chromatography (0~40% Ethyl acetate in Hexane) as a white solid.

[0243] General Procedure C: Amidation of benzylated imidazo[4,5-c]pyridine methyl ester

[0244] To a 1-dram vial were added benzylated imidazo[4,5-c]pyridine methyl ester 6d - 15d and a magnetic stir bar. The vial was sealed with a rubber septum and purged with N2for 5 minutes. The appropriate diamine (4 - 6 equiv) was added via syringe, and 10 mol% Cal2was added as amidation catalyst, the mixture was stirred at rt for 5 minutes, and then heated to 60- 90 °C for 2-30 h, until TLC indicated complete consumption of carboxylate ester 6d - 15d.The reaction was allowed to cool to room temperature, then extracted with DCM (3 * 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The free base of amide was further purified by silica gel column chromatography (0-20% Methanol in Dichloromethane) as a white solid. A portion of this material was dissolved in 0.1 M HCI in methanol (prepared from 12 M HCI (aq.)) and concentrated in vacuo. The salt was again dissolved in methanol (-1 - 4 mL) and concentrated in vacuo to remove residual water overnight to obtain the HCI salt.

[0245] methyl 4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (3a)

[0246] Using General Procedure A, histidine (20 g, 128 mmol) and 3,4-dichlorobenzaldehyde (22.56 g, 128 mmol, 1.0 equiv) were combined in aq NaOH, converted to the methyl ester, and oxidized with DMP (three step sequence i-iii described above), affording 3a as a white solid (27 g, 67% yield over 3 steps)

[0247] 1H NMR (600 MHz, cfe-DMSO) 6 13.45 (br, 1H), 9.12 (br, 1H), 8.82 (br, 1H), 8.71 (s, 1H), 8.31 (s, 1H), 7.84-7.86 (m, 1H), 3.95 (s, 3H).

[0248] 13C NMR (101 MHz, cfe-DMSO) 6 165.5, 146.3, 143.4, 139.7, 139.6, 139.2, 137.5, 131.9, 131.1, 130.6, 130.3, 128.6, 109.8, 52.3.

[0249] HRMS (ESI+) calculated for C14H10Cl2N3O2+[M+H]+: 322.0145, found: 322.0146

[0250] A / -(2-aminoethyl)-4-(3,4-dichlorophenyl)-3H-imidazo[4,5-c]pyridine-6-carboxamide dihydrochloride (3b)

[0251] Using General Procedure C above, methyl 4-(3,4-dichlorophenyl)-3 / - / -imidazo[4,5-c]pyridine-6-carboxylate 3a (50.0 mg, 0.15 mmol) and ethylenediamine (0.05 mL, d = 0.90 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 3b (35 mg, 64% yield). Subsequently 35 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.0 mL), providing 3b (as the depicted HCI salt, 38.2 mg, 98% recovery) as a white solid.

[0252] 1H NMR (500 MHz, CD3OD) 59.41 (s, 1H), 8.54 (d, J = 2.1 Hz, 1H), 8.52 (s, 1H), 8.26 (dd, J = 8.4, 2.1 Hz, 1H), 7.80 (d, J= 8.4 Hz, 1H), 3.81 (t, J = 5.9 Hz, 2H), 3.25 (t, J= 5.9 Hz, 2H).

[0253] 13C NMR (126 MHz, CD3OD) 6167.2, 146.4, 145.9, 145.4, 140.9, 137.1, 135.6, 134.2, 132.6, 132.2, 132.1, 129.9, 108.9, 41.1, 38.6.

[0254] HRMS (ESI+) calculated for C15H14Cl2N5O+[M+H]+: 350.0570, found: 350.0577

[0255] 4-(3,4-dichlorophenyl)-N-(2-(methylamino)ethyl)-3H-imidazo[4,5-c]pyridine-6-carboxamide dihydrochloride (3c)

[0256] Using General Procedure C above, methyl 4-(3,4-dichlorophenyl)-3 / - / -imidazo[4,5-c]pyridine-6-carboxylate 3a (50.0 mg, 0.15 mmol) and Af-methylethane-l^-diamine (0.07 mL, d = 0.85 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 3c (40 mg, 74% yield). Subsequently 40 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.2 mL), providing 3c (as the depicted HCI salt, 42.0 mg, 95% recovery) as a white solid.

[0257] 1H NMR (400 MHz, CD3OD) 59.29 (s, 1H), 8.60 (d, J = 2.1 Hz, 1H), 8.51 (s, 1H), 8.33 (dd, J = 8.4, 2.1 Hz, 1H), 7.79 (d, J = 8.4 Hz, 1H), 3.84 (t, J = 5.1 Hz, 2H), 3.33 (t, 2H)(overlapped with CD3OD), 2.77 (s, 3H).

[0258] 13C NMR (126 MHz, CD3OD) 6167.3, 146.4, 145.8, 145.4, 141.0, 137.0, 135.6, 134.1, 132.8, 132.2, 132.1, 130.0, 108.9, 50.8, 37.4, 33.9.

[0259] HRMS (ESI+) calculated for C16H15Cl2N5O+[M+H]+: 364.0726, found: 364.0732

[0260] 4-(3,4-dichlorophenyl)- / V-(2-(dimethylamino)ethyl)-3H-imidazo[4,5-c]pyridine-6-carboxamide dihydrochloride (3d)

[0261] Using General Procedure C above, methyl 4-(3,4-dichlorophenyl)-3H-imidazo[4,5-c]pyridine-6-carboxylate 3a (50.0 mg, 0.15 mmol) and / V. / V-dimethylethane-l,2-diamine (0.08 ml_, d = 0.81 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 3d (40 mg, 74% yield). Subsequently 40 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.2 mL), providing 3d (as the depicted HCI salt, 42.0 mg, 95% recovery) as a white solid.

[0262] 1H NMR (400 MHz, CD3OD 6 9.41 (s, 1 H), 8.54 (d, J = 2.1 Hz, 1 H), 8.52 (s, 1 H), 8.27 (dd, J = 8.4, 2.1 Hz, 1H), 7.79 (d, J = 8.4 Hz, 1H), 3.91 (t, J = 5.4 Hz, 2H), 3.48 (t, J = 5.4 Hz, 2H), 3.02 (s, 6H).

[0263] 13C NMR (126 MHz, CD3OD) 5 167.3, 146.4, 145.6, 145.5, 141.1, 137.2, 135.5, 134.1, 133.3, 132.2, 132.0, 130.0, 108.9, 58.7, 44.0, 36.2.

[0264] HRMS (ESI+) calculated for Ci7Hi7CI2N5O+[M+H]+: 378.0883, found: 378.0885

[0265] methyl 4-(3,4-dichlorophenyl)-1-methyl-1H-imidazo[4,5-c]pyridine-6-carboxylate (4a)

[0266] To a 20 mL vial methyl 4-(3,4-dichlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate (3a) (200.0 mg, 0.6 mmol) and cesium carbonate (303 mg, 0.9 mmol, 1.5 equiv), methyl iodide (0.08 mL, d = 2.28 g / mL, 1.24 mmol, 2 equiv) was added dropwise in DMF at room temperature. The vial was capped and stirred at room temperature for 24h. TLC typicallyindicated complete consumption of the oxidation intermediate. The reaction mixture was extracted with EtOAc (3 x 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 4a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (130 mg, 62%).

[0267] 1H NMR (400 MHz, d6-DMSO) 5 9.09 (d, J = 2.0 Hz, 1H), 8.81 (dd, J = 8.5, 2.0 Hz, 1H), 8.67 (s, 1H), 8.45 (s, 1H), 7.86 (d, J = 8.5 Hz, 1H), 4.01 (s, 3H), 3.96 (s, 3H).

[0268] 13C NMR (101 MHz, cfe-DMSO) 5 165.4, 148.7, 143.4, 140.8, 139.7, 139.1, 137.3, 132.0, 131.1, 130.6, 130.2, 128.6, 108.7, 52.4, 31.3.

[0269] HRMS (ESI+) calculated for C15Hi2CI2N3O2+[M+H]+: 336.0301, found: 336.0305

[0270] N-(2-aminoethyl)-4-(3,4-dichlorophenyl)-1-rnethyl-1H-irnidazo[4,5-c]pyridine-6-carboxamide hydrochloride (4b)

[0271] Using General Procedure C above, methyl 4-(3,4-dichlorophenyl)-1-methyl-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 4a (50.0 mg, 0.15 mmol) and ethylenediamine (0.05 mL, d = 0.90 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 4b (32 mg, 59% yield). Subsequently 32 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.85 mL), providing 4b (as the depicted HCI salt, 30 mg, 84% recovery) as a white solid.

[0272] 1H NMR (600 MHz, DMSO) 5 9.31 (d, J = 2.1 Hz, 1H), 9.17 (t, J = 6.3 Hz, 1H), 9.07 (dd, J = 8.5, 2.1 Hz, 1H), 8.63 (s, 1H), 8.37 (s, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.43 (br, 3H), 4.00 (s, 3H), 3.64 (q, J = 6.3 Hz, 2H), 3.03 (t, J = 6.3 Hz, 2H).

[0273] 13C NMR (151 MHz, DMSO) 0 164.9, 148.3, 142.6, 142.1, 141.5, 139.4, 137.3, 132.1, 131.3, 130.7, 130.5, 129.3, 105.5, 40.0, 37.7, 31.3.

[0274] HRMS (ESI+) calculated for C16Hi5Cl2N5O+[M+H]+: 364.0726, found: 364.0733

[0275] methyl 1-butyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (5a)

[0276] To a 20 mL vial methyl 4-(3,4-dichlorophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxylate (3a) (200.0 mg, 0.6 mmol) and cesium carbonate (303 mg, 0.9 mmol, 1.5 equiv), n-butyl bromide (0.15 mL, d = 1.27 g / mL, 1.24 mmol, 2 equiv) was added dropwise in DMF at room temperature. The vial was capped and stirred at 60 °C for 24h. TLC typically indicated complete consumption of the oxidation intermediate. The reaction mixture was extracted with EtOAc (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 5a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (189 mg, 80%).

[0277] 1H NMR (400 MHz, DMSO) 59.08 (d, J= 2.0 Hz, 1H), 8.80 (dd, J= 8.5, 2.0 Hz, 1H), 8.75 (s, 1H), 8.48 (s, 1H), 7.86 (d, J = 8.5 Hz, 1H), 4.44 (t, J= 7.2 Hz, 2H), 3.96 (s, 3H), 1.81 (5-let, J = 7.2 Hz, 2H), 1.29 (6-let, J = 7.2 Hz, 3H), 0.91 (t, J = 7.2 Hz, 3H).

[0278] 13C NMR (151 MHz, DMSO) 0 165.5, 148.3, 143.8, 140.3, 139.9, 139.4, 137.4, 132.1, 131.2, 130.8, 130.3, 128.8, 108.8, 52.5, 44.5, 31.6, 19.3, 13.4.

[0279] HRMS (ESI+) calculated for Ci8H18Cl2N3O2+[M+H]+: 378.0771, found: 378.0771

[0280] N-(2-aminoethyl)-1-butyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (5b)

[0281] Using General Procedure C above, methyl 1 -butyl-4-(3,4-dichlorophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxylate 5a (50.0 mg, 0.15 mmol) and ethylenediamine (0.05 mL,d = 0.90 g / mL, 0.74 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 4b (38 mg, 70% yield). Subsequently 38 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.85 ml_), providing 4b (as the depicted HCI salt, 39 mg, 99% recovery) as a white solid.

[0282] 1H NMR (600 MHz, DMSO) 6 9.31 (d, J = 2.1 Hz, 1H), 9.17 (t, J = 6.3 Hz, 1H), 9.07 (dd, J = 8.5, 2.1 Hz, 1H), 8.71 (s, 1H), 8.40 (s, 1H), 7.83 (d, J= 8.5 Hz, 1H), 7.57 (s, 3H), 4.44 (t, J = 6.4 Hz, 2H), 3.64 (q, J = 6.4 Hz, 2H), 3.04 (t, J = 6.4 Hz, 2H), 1.81 (5-let, J = 7.4 Hz 2H), 1.28 (s, J = 7.4 Hz 2H), 0.90 (t, J = 7.4 Hz, 3H).

[0283] 13C NMR (151 MHz, DMSO) 5 164.9, 147.8, 142.9, 142.2, 140.8, 139.5, 137.3, 132.1, 131.3, 130.8, 130.5, 129.3, 105.5, 44.4, 39.9, 37.6, 31.6, 19.3, 13.4.

[0284] HRMS (ESI+) calculated for C19H22CI2N5O+[M+H]+: 406.1196, found: 406.1201

[0285] methyl 1-benzyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (6a)

[0286] Using General Procedure B above, methyl 4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (3a) (5g, 15.52 mmol) and benzyl bromide (3.7 mL, d = 1.44 g / mL, 31.04 mmol, 2 equiv) and cesium carbonate (7.59g, 24 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 6a was further purified by silica gel column chromatography (0~40% Ethyl acetate in Hexane) and isolated as a white solid (4.8 g, 82%).

[0287] 1H NMR (400 MHz, CDCh) 58.89 (d, J = 2.0 Hz, 1H), 8.65 (dd, J= 8.5, 2.0 Hz, 1H), 8.19 (s, 1H), 8.17 (s, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.44 - 7.33 (m, 3H), 7.23 - 7.14 (m, 2H), 5.46 (s, 2H), 4.03 (s, 3H).

[0288] 13C NMR (151 MHz, CDCh) 6 166.4, 147.0, 146.2, 140.9, 140.7, 140.5, 137.1, 134.2, 133.8, 132.8, 131.4, 130.5, 129.6, 129.1, 128.9, 127.3, 107.9, 53.1, 49.4.

[0289] HRMS (ESI+) calculated for C21H16Cl2N3O2+[M+H]+: 412.0614, found: 412.0614

[0290] N-(2-aminoethyl)-1-benzyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (6b)

[0291] Using General Procedure C above, methyl 1-benzyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate 6a (50.0 mg, 0.12 mmol) and ethylenediamine (0.05 mL, d = 0.90 g / mL, 0.6 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 6b (33 mg, 62% yield). Subsequently 33 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.85 mL), providing 6b (as the depicted HCI salt, 36 mg, 100% recovery) as a white solid.

[0292] 1H NMR (400 MHz, CD3OD 0 9.27 (t, J = 6.3 Hz, 1H), 9.04 (d, J = 2.0 Hz, 2H), 8.78 (dd, J = 8.5, 2.0 Hz, 2H), 8.63 (s, 2H), 8.27 (s, 2H), 7.70 (d, J = 8.5 Hz, 2H), 7.52 - 7.22 (m, 5H), 5.65 (s, 2H), 3.78 (t, J = 6.3 Hz, 2H), 3.22 (t, J = 6.3 Hz, 2H).

[0293] 13C NMR (126 MHz, CD3OD) 6168.1, 148.7, 146.3, 143.7, 142.1, 140.3, 138.2, 136.6, 134.8, 133.6, 132.4, 131.6, 130.3, 130.2, 129.6, 128.7, 106.9, 41.2, 38.6, 30.8.

[0294] HRMS (ESI+) calculated for C22H20Cl2N5O+[M+H]+: 440.1039, found: 440.1042

[0295] 1-benzyl-4-(3,4-dichlorophenyl)-N-(2-(methylamino)ethyl)-1 H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (6c)

[0296] Using General Procedure C above, methyl 1-benzyl-4-(3,4-dichlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 6a (50.0 mg, 0.12 mmol) and 1T -methylethane- 1,2-diamine (0.07 mL, d = 0.85 g / mL, 0.71 mmol, 5 equiv) were combined and heated to 90 °C for20 h, affording the free base of 6c (45 mg, 82% yield). Subsequently 45 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.96 ml_), providing 6c (as the depicted HCI salt, 48 mg, 100% recovery) as a white solid.

[0297] 1H NMR (500 MHz, CD3OD) 69.04 (s, 1H), 8.82 (d, J= 2.1 Hz, 1H), 8.55 (dd, J = 8.4, 2.1 Hz, 1H), 8.35 (s, 1H), 7.72 (d, J= 8.4 Hz, 1H), 7.48 - 7.30 (m, 5H), 5.72 (s, 2H), 3.81 (d, J = 6.2 Hz, 2H), 3.29 (d, J = 6.2 Hz, 2H), 2.75 (s, 3H).

[0298] 13C NMR (126 MHz, CD3OD) 6 167.6, 148.5, 146.1, 144.3, 141.8, 138.1, 137.6, 136.1, 135.1, 133.7, 132.3, 131.7, 130.3, 130.2, 129.8, 128.9, 107.2, 50.9, 50.8, 37.4, 33.9.

[0299] HRMS (ESI+) calculated for C23H22CI2N5O+[M+H]+: 454.1196, found: 454.1197

[0300] 1-benzyl-4-(3,4-dichlorophenyl)- / V-(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (6d)

[0301] Using General Procedure C above, methyl 1-benzyl-4-(3,4-dichlorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 6a (50.0 mg, 0.12 mmol) and Af. AT-dimethylethane-l,2-diamine (0.08 mL, d = 0.81 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 6d (49 mg, 88% yield). Subsequently 49 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.1 mL), providing 6d (as the depicted HCI salt, 50 mg, 95% recovery) as a white solid.

[0302] 1H NMR (500 MHz, CD3OD) 68.91 (d, J = 2.1 Hz, 1H), 8.88 (s, 1H), 8.66 (dd, J = 8.5, 2.1 Hz, 1H), 8.32 (s, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.44 - 7.32 (m, 5H), 5.69 (s, 2H), 3.88 (t, J = 5.8 Hz, 2H), 3.44 (t, J = 5.8 Hz, 2H), 2.99 (s, 6H).

[0303] 13C NMR (126 MHz, CD3OD) 6 167.9, 148.7, 146.2, 143.8, 141.9, 139.5, 137.9, 136.4, 134.9, 133.6, 132.4, 131.6, 130.3, 129.7, 128.7, 107.1, 58.9, 50.5, 43.7, 36.2. Proposed accidental equivalence for two aromatic carbons.

[0304] HRMS (ESI+) calculated for C24H24CI2N5O+[M+H]+: 468.1352, found: 468.1359

[0305] A / -(1 -amino-2-methylpropan-2-yl)-1 -benzyl-4-(3,4-dichlorophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (6e)

[0306] Using General Procedure C above, methyl 1-benzyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate 6a (50.0 mg, 0.12 mmol) and tert-butyl (2-amino-2-methylpropyl)carbamate (114mg, 0.61 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the terminal N-BOC protected free base of 6e (55 mg, 96% yield). The free base was then obtained by stirring with TFA in DCM for 5h to obtain the free base of 6e. (45 mg, 100% conversion). Subsequently 45 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.97 ml_), providing 6e (as the depicted HCI salt, 48 mg, 99% recovery) as a white solid.

[0307] 1H NMR (600 MHz, DMSO) 5 9.16 (d, J = 2.1 Hz, 1H), 8.91 (s, 1H), 8.83 (dd, J = 8.5, 2.1 Hz, 1H), 8.29 (s, 1H), 8.00 (t, J = 5.9 Hz, 3H), 7.87 (d, J = 8.5 Hz, 1H), 7.49 - 7.25 (m, 5H), 5.72 (s, 2H), 3.31 (q, J = 5.9 Hz, 2H), 1.49 (s, 6H).

[0308] 13C NMR (151 MHz, DMSO) 5 163.9, 148.3, 142.8, 142.3, 140.7, 139.7, 136.9, 136.2, 132.3, 131.3, 130.8, 130.7, 128.9, 128.9, 128.1, 127.4, 105.4, 51.9, 48.1, 45.2, 24.9.

[0309] HRMS (ESI+) calculated for C24H24Cl2N5O+[M+H]+: 468.1352, found: 468.1357

[0310] N-(2-amino-2-methylpropyl)-1 -benzyl-4-(3,4-dichlorophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (6f)

[0311] Using General Procedure C above, methyl 1-benzyl-4-(3,4-dichlorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 6a (50.0 mg, 0.12 mmol) and tert-butyl (1-amino-2-methylpropan-2-yl)carbamate (114mg, 0.61 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the terminal N-BOC protected free base of 6f (50 mg, 72% yield). The free base was then obtained by stirring with TFA in DCM for 5h to obtain the free base of 6f (41 mg, 100% conversion). Subsequently 41 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.97 ml_), providing 6f (as the depicted HCI salt, 44 mg, 99% recovery) as a white solid.

[0312] 1H NMR (600 MHz, DMSO) 5 9.30 (d, J = 2.1 Hz, 1H), 9.18 (t, J = 6.7 Hz, 1H), 9.04 (dd, J= 8.5, 2.1 Hz, 1H), 8.89 (s, 1H), 8.31 (s, 1H), 8.01 (s, 3H), 7.84 (d, J= 8.5 Hz, 1H), 7.60 - 7.21 (m, 5H), 5.74 (s, 3H), 3.55 (m, 2H)(overlapped with water peak), 1.29 (s, 6H).

[0313] 13C NMR (151 MHz, DMSO) 5 165.2, 148.2, 143.1, 142.2, 140.7, 139.8, 137.1, 136.3, 132.2, 131.3, 130.9, 130.6, 129.3, 128.9, 128.1, 127.3, 105.9, 54.8, 47.9, 46.7, 23.7.

[0314] HRMS (ESI+) calculated for C24H24Cl2N5O+[M+H]+: 468.1352, found: 468.1357

[0315] 1-benzyl-A / -butyl-4-(3,4-dichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide (6g)

[0316] Using General Procedure C above, methyl 1-benzyl-4-(3,4-dichlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 6a (50.0 mg, 0.12 mmol) and n-Butylamine (0.06 mL, d = 0.74 g / mL, 0.6 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 6g (50 mg, 90% yield) as a white solid.

[0317] 1H NMR (400 MHz, DMSO) 69.27 (d, J= 2.1 Hz, 1H), 9.01 (dd, J= 8.5, 2.1 Hz, 1H), 8.95 (t, J = 6.2 Hz, 1H), 8.83 (s, 1H), 8.26 (s, 1H), 7.82 (d, J = 8.5 Hz, 1H), 7.48 - 7.23 (m, 5H), 5.71 (s, 2H), 3.36 (q, J = 6.9 Hz, 2H), 1.56 (5-let, J = 6.9 Hz, 2H), 1.34 (6-let, J = 7.3 Hz, 2H), 0.92 (t, J= 7.3 Hz, 3H).

[0318] 13C NMR (101 MHz, DMSO) 5163.8, 147.8, 142.9, 142.87, 140.7, 139.5, 137.2, 136.2, 132.1, 131.2, 130.8, 130.5, 129.3, 128.8, 128.0, 127.3, 105.5, 47.9, 38.7, 31.6, 19.7, 13.8.

[0319] HRMS (ESI+) calculated for C24H22Cl2N4O+[M+H]+: 453.1243, found: 453.1251

[0320] methyl 1-benzyl-4-(4-chloro-3-fluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (7a)

[0321] Using General Procedure B above, methyl 4-(3,4-difluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.5 mmol) and benzyl bromide (0.74 ml_, d = 1.44 g / mL, 6.91 mmol, 2 equiv) and cesium carbonate (1.69 g, 5.2 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 7a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (641 mg, 49%).

[0322] 1H NMR (400 MHz, DMSO) 58.86 (s, 1H), 8.79 (ddd, J = 12.7, 8.3, 2.2 Hz, 1H), 8.70 -8.64 (m, 1H), 8.34 (s, 1H), 7.62 (dt, J= 10.7, 8.6 Hz, 1H), 7.41 -7.27 (m, 6H), 5.71 (s, 2H), 3.92 (s, 3H).

[0323] 13C NMR (101 MHz, DMSO) 5 165.4, 149.2 (d,1JCF= 244.15 Hz), 148.6 (d,1JCF= 249.42 Hz), 148.3, 144.3, 140.1, 139.8, 139.4, 136.2, 134.3 (d,3JCF= 5.1 Hz), 128.9, 128.0, 127.4, 125.8, 117.5 (d,2JCF= 18.0 Hz), 108.5, 52.4, 47.9. Proposed accidental equivalence for two aromatic carbons

[0324] 19F NMR (376 MHz, CDCI3) 6 -136.69 (d, J= 22.4 Hz), -137.92 (d, J = 22.4 Hz).

[0325] HRMS (ESI+) calculated for C21H16Cl2N3O2+[M+H]+: 380.1205, found: 380.1214

[0326] A / -(2-aminoethyl)-1-benzyl-4-(3,4-difluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (7b)

[0327] Using General Procedure C above, methyl 1-benzyl-4-(3,4-difluorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 7a (50.0 mg, 0.13 mmol) and ethylenediamine (0.05 mL, d = 0.90 g / mL, 0.6 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 7b (31 mg, 57% yield). Subsequently 31 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.75 mL), providing 7b (as the depicted HCI salt, 33 mg, 99% recovery) as a white solid.

[0328] 1H NMR (600 MHz, DMSO) 5 9.17 (t, J = 6.2 Hz, 1H), 9.09 (ddd, J = 12.8, 8.06, 1.9 Hz, 1H), 9.02 (ddd, J= 10.2, 4.8, 1.9 Hz, 1H), 8.87 (s, 1H), 8.27 (s, 1H), 8.05 (d, J = 23.1 Hz, 3H), 7.63 (dt, J = 10.5, 8.6 Hz, 1 H), 7.44 - 7.27 (m, 6H), 5.72 (s, 2H), 3.64 (q, J = 6.2 Hz, 2H), 3.03 (s, J = 6.2, 2H).

[0329] 13C NMR (151 MHz, DMSO) 6 164.8, 149.9 (dd,1JCF= 243.99 Hz,2JCF= 12.94 Hz), 147.9, 143.3, 142.2, 140.6, 139.5, 136.3, 134.1, 128.9, 128.0, 127.3, 126.7, 126.4, 121.1 (dd,1JCF= 250.08 Hz,2JCF= 12.94 Hz), 117.9 (d,2JCF = 19.28 Hz), 117.3 (d,2JCF = 17.65 Hz), 105.4, 47.9, 38.7, 36.9.

[0330] 19F NMR (376 MHz, DMSO) 6 -137.24 (d,3JFF = 22.4 Hz, 1F), -138.45 (d,3JFF = 22.4 Hz, 1F).

[0331] HRMS (ESI+) calculated for C22H20F2N5O+[M+H]+: 408.1630, found: 408.1633

[0332] 1-benzyl-4-(3,4-difluorophenyl)-N-(2-(methylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (7c)

[0333] Using General Procedure C above, methyl 1-benzyl-4-(3,4-difluorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 7a (50.0 mg, 0.13 mmol) and A / 1-methylethane- 1,2-diamine (0.07 mL, d = 0.85 g / mL, 0.71 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 7c (23 mg, 42% yield). Subsequently 23 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.55 mL), providing 7c (as the depicted HCI salt, 25 mg, 100% recovery) as a white solid.

[0334] 1H NMR (600 MHz, MeOD) 5 8.80 (ddd, J = 12.5, 8.2, 2.2 Hz, 1H), 8.72-8.65 (m, 1H), 8.59 (s, 1H), 8.21 (s, 1H), 7.42 (dt, J = 12.5, 8.2 Hz, 1H), 7.34 (m, 5H), 5.62 (s, 2H), 3.79 (t, J = 6.45 Hz 2H), 3.24 ((t, J = 6.45 Hz 2H), 2.71 (s, 3H).

[0335] 13C NMR (151 MHz, MeOD) 5 168.3, 152.5 (dd,1JCF= 250.38 Hz,2JCF= 13.26 Hz), 148.7, 146.5, 143.2, 142.1, 141.3, 136.9, 135.6, 130.2, 129.5, 128.6, 127.6 (dd, J = 6.6, 3.5 Hz), 121.5 (dd,1JCF = 245.24 Hz,2JCF= 12.45 Hz) 119.5 (d,2JCF= 19.19 Hz), 118.1 (d,2JCF= 17.57 Hz), 106.4, 51.1, 50.0, 37.7, 34.1.

[0336] 19F NMR (376 MHz, DMSO) 0 -140.00 (d,3JFF = 20.6 Hz, 1F), -141.35 (d,3JFF = 20.6 Hz, 1F).

[0337] HRMS (ESI+) calculated for C23H22F2N5O+[M+H]+: 422.1787, found: 422.1793

[0338] 1-benzyl-4-(3,4-difluorophenyl)-A / -(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (7d)

[0339] Using General Procedure C above, methyl 1-benzyl-4-(3,4-difluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate 7a (50.0 mg, 0.13 mmol) and AA. AA-dimethylethane-l,2-diamine (0.08 mL, d = 0.81 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 7d (43 mg, 80% yield). Subsequently 43 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.9 mL), providing 7d (as the depicted HCI salt, 46 mg, 100% recovery) as a white solid.

[0340] 1H NMR (600 MHz, MeOD) 6 8.82 (ddd, J = 12.5, 8.5, 2.2 Hz, 1H), 8.78 - 8.71 (m, 1H), 8.62 (s, 1H), 8.24 (s, 1H), 7.43 (dt, J = 10.4, 8.5 Hz, 1H), 7.40 - 7.31 (m, 5H), 5.64 (s, 2H), 3.88 (t, J = 6.2 Hz, 2H), 3.44 (t, J = 6.2 Hz, 2H), 2.99 (s, 6H).

[0341] 13C NMR (151 MHz, MeOD) 6 168.4, 152.5 (dd,1JCF= 249.56 Hz,2JCF= 12.93 Hz), 151.5 (dd,1JCF= 245.3 Hz,2JCF= 12.65 Hz), 148.8, 146.6, 143.1, 142.2, 141.4, 136.9, 135.6, 130.2, 129.5, 128.6, 127.6, 119.5 (d,2JCF= 19.2 Hz), 118.1 (d,2JCF= 17.6 Hz), 106.5, 59.0, 50.1, 43.9, 36.2.

[0342] 19F NMR (376 MHz, DMSO) 6 -139.84 (d,3JFF= 20.7 Hz, 1F), -141.23 (d,3JFF= 20.7 Hz, 1F).

[0343] HRMS (ESI+) calculated for C24H24F2N5O+[M+H]+: 436.1943, found: 436.1948

[0344] methyl 1-benzyl-4-(3-chloro-4-fluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (8a)

[0345] Using General Procedure B above, methyl 4-(3-chloro-4-fluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.27 mmol) and benzyl bromide (0.77 mL, d = 1.44 g / mL, 6.91 mmol, 2 equiv) and cesium carbonate (1.60 g, 4.91 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 8a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (425 mg, 33%).

[0346] 1H NMR (400 MHz, DMSO) 6 8.9 (dd, J = 7.5, 2.2 Hz, 1H), 8.8 (s, 1H), 8.8 (ddd, J = 8.8, 4.9, 2.2 Hz, 1 H), 8.3 (s, 1 H), 7.6 (t, J = 9.0 Hz, 1 H), 7.4 - 7.2 (m, 5H), 5.7 (s, 2H), 3.9 (s, 3H).

[0347] 13C NMR (101 MHz, DMSO) 5 165.4, 157.9 (d,1JCF= 250.0 Hz), 148.2, 144.1, 140.0, 139.7, 139.4, 136.1, 134.4 (d, J= 3.4 Hz), 130.7, 129.5 (d,3JCF= 7.7 Hz), 128.8, 128.0, 127.4, 119.5 (d,2JCF= 17.9 Hz), 116.8 (d,2JCF= 21.0 Hz), 108.4, 52.4, 47.9.

[0348] 19F NMR (376 MHz, CDCI3) 6 -114.20.

[0349] HRMS (ESI+) calculated for C21H16ClFN3O2+[M+H]+: 396.0910, found: 396.0919

[0350] N-(2-aminoethyl)-1-benzyl-4-(3-chloro-4-fluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (8b)

[0351] Using General Procedure C above, methyl 1-benzyl-4-(3-chloro-4-fluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 8a (50.0 mg, 0.12 mmol) and ethylenediamine (0.05 mL, d = 0.90 g / mL, 0.5 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 8b (40 mg, 72% yield). Subsequently 31 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.75 mL), providing 8b (as the depicted HCI salt, 33 mg, 99% recovery) as a white solid.

[0352] 1H NMR (600 MHz, DMSO) 59.26 (dd, J= 7.5, 2.2 Hz, 1H), 9.17 (t, J= 6.1 Hz, 1H), 9.10 (ddd, J= 8.9, 4.9, 2.2 Hz, 1H), 8.87 (s, 1H), 8.06 (s, 3H), 7.61 (t, J= 8.9 Hz, 1H), 7.43-7.23 (m, 5H), 5.73 (s, 3H), 3.64 (q, J = 6.2 Hz, 2H), 3.03 (s, J = 6.2 Hz, 2H).

[0353] 13C NMR (151 MHz, DMSO) 5164.9, 157.9 (d,1JCF= 249.88 Hz), 147.9, 143.3, 142.3, 140.6, 139.5, 136.3, 134.4 (d,4JCF = 3.6 Hz), 131.2, 130.2 (d,3JCF = 7.5 Hz), 128.9, 128.0, 127.3, 119.6 (d,2JCF = 17.6 Hz), 116.8 (d,2JCF= 20.7 Hz), 105.4, 47.9, 38.7, 36.9.

[0354] 19F NMR (376 MHz, DMSO) 6 -115.00.

[0355] HRMS (ESI+) calculated for C22H20FClN5O+[M+H]+: 424.1335, found: 424.1341

[0356] 1-benzyl-4-(3-chloro-4-fluorophenyl)-N-(2-(methylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (8c)

[0357] Using General Procedure C above, methyl 1-benzyl-4-(3-chloro-4-fluorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 8a (50.0 mg, 0.12 mmol) and A / 1-methylethane- 1,2-diamine (0.07 mL, d = 0.85 g / mL, 0.71 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 8c (31 mg, 56% yield). Subsequently 31 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.66 mL), providing 8c (as the depicted HCI salt, 31 mg, 92% recovery) as a white solid.

[0358] 1H NMR (600 MHz, MeOD) 68.95 (dd, J = 7.4, 2.2 Hz, 1H), 8.79 (ddd, J = 8.7, 4.8, 2.2 Hz, 1H), 8.59 (s, 1H), 8.19 (s, 1 H), 7.45 - 7.26 (m, 5H), 5.60 (s, 3H), 3.82 (dd, J = 6.2, 5.1 Hz, 2H), 3.29 (dd, J = 6.2, 5.1 Hz, 2H), 2.75 (s, 3H).

[0359] 13C NMR (151 MHz, MeOD) 6 168.3, 160.1 (d,1JCF= 251.48 Hz), 159.3, 148.7, 146.4, 143.2, 142.0, 141.2, 136.8, 135.6 (d,4JCF =3.8 Hz), 132.8, 131.2 (d,3JCF = 7.6 Hz), 130.2, 129.5, 128.6, 117.4 (d,2JCF= 21.23 Hz), 106.4, 50.9, 50.1, 37.4, 33.9.

[0360] 19F NMR (376 MHz, DMSO) 0 -116.21 - -116.81 (m).

[0361] HRMS (ESI+) calculated for C23H22FClN5O+[M+H]+: 438.1491, found: 438.1495

[0362] 1-benzyl-4-(3-chloro-4-fluorophenyl)-A / -(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (8d)

[0363] Using General Procedure C above, methyl 1-benzyl-4-(3-chloro-4-fluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 8a (50.0 mg, 0.12 mmol) and AA. Af-dimethylethane-l ^-diamine (0.08 mL, d = 0.81 g / mL, 0.78 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 8d (39 mg, 71% yield). Subsequently 39 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.66 mL), providing 8d (as the depicted HCI salt, 42 mg, 99% recovery) as a white solid.

[0364] 1H NMR (600 MHz, DMSO) 0 9.94 (s, 1H), 9.26 (dd, J = 7.5, 2.2 Hz, 1H), 9.22 (t, J = 6.2 Hz, 1H), 9.10 (ddd, J = 8.8, 4.9, 2.2 Hz, 1H), 8.88 (s, 1H), 8.27 (s, 1 H), 7.62 (t, J = 9.0 Hz, 1 H), 7.47 - 7.24 (m, 5H), 5.73 (s, 2H), 3.74 (q, J = 6.2 Hz, 2H), 3.31 (q, J = 6.2 Hz)(overlapped with water), 2.84 (s, 6H).

[0365] 13C NMR (151 MHz, DMSO) 5 164.9, 157.8 (d,1JCF= 248.8 Hz), 157.1, 148.1, 143.3, 142.1, 140.6, 139.54, 136.3, 134.4 (d,4JCF =3.4 HZ), 131.2, 130.2 (d,3JCF = 7.7 Hz), 128.9, 128.1, 127.3, 119.6 (d,2JCF= 17.8 Hz), 116.8 (d,2JCF= 20.5 Hz), 105.5, 56.2, 47.9, 42.5, 34.6.

[0366] 19F NMR (376 MHz, DMSO) 6 -114.93.

[0367] HRMS (ESI+) calculated for C24H24FClN5O+[M+H]+: 452.1648, found: 452.1649

[0368] methyl 1-benzyl-4-(4-chloro-3-fluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (9a)

[0369] Using General Procedure B above, methyl 4-(4-chloro-3-fluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.27 mmol) and benzyl bromide (0.77 mL, d = 1.44 g / mL, 6.91 mmol, 2 equiv) and cesium carbonate (1.60 g, 4.91 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 * 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 9a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (593 mg, 46%).

[0370] 1H NMR (400 MHz, DMSO) 68.86 (s, 1 H), 8.76 (dd, J = 11.4, 1.9 Hz, 1 H), 8.66 (dd, J = 8.5, 1.9 Hz, 1H), 8.35 (s, 1H), 7.75 (t, J= 8.5 Hz, 1H), 7.41 -7.24 (m, 5H), 5.71 (s, 2H), 3.92 (s, 3H).

[0371] 13C NMR (101 MHz, DMSO) 0 165.4, 157.1 (d,1JCF= 245.1 Hz), 148.3, 144.0, 140.1, 140.0, 139.4, 137.7 (d,3JcF = 7.2 Hz), 136.1, 130.6, 128.9, 128.0, 127.4, 125.8, 120.6 (d,2JCF = 17.7 Hz), 116.5 (d,2JCF = 23.0 Hz), 108.8, 52.4, 48.0.

[0372] 19F NMR (376 MHz, CDCI3) 6 -115.66.

[0373] HRMS (ESI+) calculated for C21H16ClFN3O2+[M+H]+: 396.0910, found: 396.0912

[0374] N-(2-aminoethyl)-1-benzyl-4-(4-chloro-3-fluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (9b)

[0375] Using General Procedure C above, methyl 1-benzyl-4-(4-chloro-3-fluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 9a (50.0 mg, 0.12 mmol) and ethylenediamine (0.05 mL, d = 0.90 g / mL, 0.5 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 9b (45 mg, 81% yield). Subsequently 45 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.66 mL), providing 9b (as the depicted HCI salt, 46 mg, 96% recovery) as a white solid.

[0376] 1H NMR (600 MHz, DMSO) 59.18 (t, J= 6.2 Hz, 1H), 9.05 (ddd,, J= 16.6, 11.4, 2.2 Hz, 1H), 8.88 (s, 1H), 8.29 (s, 1H), 8.03 (s, 3H), 7.79 (t, J= 8.2 Hz, 1H), 7.44 - 7.26 (m, 5H), 5.73 (s, 2H), 3.64 (q, J = 6.2 Hz, 2H), 3.04 (br, 2H).

[0377] 13C NMR (151 MHz, DMSO) 5 164.8, 157.4 (d,1JCF= 244.7 Hz), 148.0, 143.1, 142.2, 140.7, 139.7, 137.8 (d,3JCF = 7.2 Hz), 136.3, 130.5, 128.9, 128.0, 127.3, 126.6, 120.7 (d,2JCF = 17.7 Hz), 116.9 (d,2JCF = 22.8 Hz), 105.7, 47.9, 38.7, 36.9.

[0378] 19F NMR (376 MHz, DMSO) 5 -116.08.

[0379] HRMS (ESI+) calculated for C24H24FClN5O+[M+H]+: 424.1335, found: 424.1338

[0380] 1-benzyl-4-(4-chloro-3-fluorophenyl)-N-(2-(methylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (9c)

[0381] Using General Procedure C above, methyl 1-benzyl-4-(4-chloro-3-fluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 9a (50.0 mg, 0.12 mmol) and A / 1-methylethane- 1,2-diamine (0.07 mL, d = 0.85 g / mL, 0.71 mmol, 5 equiv) were combined and heated to 90 °C for20 h, affording the free base of 9c (37 mg, 67% yield). Subsequently 37 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.78 ml_), providing 9c (as the depicted HCI salt, 39 mg, 99% recovery) as a white solid.

[0382] 1H NMR (600 MHz, DMSO) 6 9.21 (t, J = 6.2 Hz, 1H), 9.05 (ddd, J = 16.6, 11.4, 2.2 Hz, 1 H), 8.89 (s, 1 H), 8.77 (br, 2H), 8.28 (s, 1 H), 7.79 (t, J = 8.2 Hz, 1 H), 7.47 - 7.21 (m, 5H), 5.73 (s, 2H), 3.68 (q, J = 6.2 Hz, 2H), 3.21 - 3.08 (m, 2H), 2.65 - 2.55 (m, 3H).

[0383] 13C NMR (151 MHz, DMSO) 6 164.9, 157.4 (d,1JCF = 244.7 Hz), 148.1, 143.1, 142.1, 140.7, 139.7, 137.8 (d,3JCF = 7.2 Hz), 136.3, 130.5, 128.9, 128.0, 127.3, 126.6, 120.7 (d,2JCF = 17.6 Hz), 116.9 (d,2JCF = 22.6 Hz), 105.7, 48.2, 47.9, 35.6, 32.7.

[0384] 19F NMR (376 MHz, DMSO) 6 -116.07.

[0385] HRMS (ESI+) calculated for C23H22FClN5O+[M+H]+: 438.1491, found: 438.1502

[0386] 1-benzyl-4-(4-chloro-3-fluorophenyl)-N-(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (9d)

[0387] Using General Procedure C above, methyl 1-benzyl-4-(4-chloro-3-fluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 9a (50.0 mg, 0.12 mmol) and Af. AT-dimethylethane-l,2-diamine (0.07 mL, d = 0.81 g / mL, 0.72 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 9d (33 mg, 60% yield). Subsequently 37 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.71 mL), providing 9d (as the depicted HCI salt, 35 mg, 99% recovery) as a white solid.

[0388] 1H NMR (600 MHz, DMSO) 6 9.79 (br, 1H), 9.21 (t, J = 6.2 Hz, 1H), 9.05 (ddd, J = 12.4, 11.2, 2.1 Hz, 1H), 8.89 (s, 1H), 8.28 (s, 1H), 7.80 (t, J = 8.1 Hz, 1 H), 7.52 - 7.21 (m, 5H), 5.73 (s, 2H), 3.72 (q, J = 6.2 Hz, 2H), 3.27 (br, 2H), 2.81 (s, 6H).

[0389] 13C NMR (151 MHz, DMSO) 6 164.8, 157.4 (d,1JCF= 244.6 Hz), 148.1, 143.1, 142.1, 140.7, 139.8, 137.8 (d,3JCF= 7.4 Hz), 136.3, 130.5, 128.9, 128.1, 127.3, 126.6, 120.7 (d,2JCF= 17.7 Hz), 116.9 (d,2JCF = 22.6 Hz), 105.8, 56.3, 47.9, 42.7, 34.7.

[0390] 19F NMR (376 MHz, DMSO) 6 -116.04.

[0391] HRMS (ESI+) calculated for C24H24FClN5O+[M+H]+: 452.1648, found: 452.1648

[0392] methyl 1-benzyl-4-(3,4,5-trichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (10a)

[0393] Using General Procedure B above, methyl 4-(3,4,5-trichlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 2.8 mmol) and benzyl bromide (0.66 ml_, d = 1.44 g / mL, 5.61 mmol, 2 equiv) and cesium carbonate (1.37 g, 4.21 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 10a was further purified by silica gel column chromatography (0~40% Ethyl acetate in Hexane) and isolated as a white solid (300 mg, 24%).

[0394] 1H NMR (400 MHz, DMSO) 0 8.9 (s, 1H), 8.9 (s, 2H), 8.3 (s, 1H), 7.4 - 7.2 (m, 3H), 5.7 (s, 1H), 3.9 (s, 2H), 3.3 (s, 3H).

[0395] 13C NMR (101 MHz, DMSO) 5 165.1, 148.6, 142.1, 140.2, 140.0, 139.3, 137.1, 136.0, 133.0, 130.5, 128.9, 128.6, 128.1, 127.4, 109.1, 52.5, 48.0.

[0396] HRMS (ESI+) calculated for C21H15Cl3N3O2+[M+H]+: 446.0224, found: 446.0238

[0397] N-(2-aminoethyl)-1-benzyl-4-(3,4,5-trichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (10b)

[0398] Using General Procedure C above, methyl 1-benzyl-4-(3,4,5-trichlorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 10a (30.0 mg, 0.07 mmol) and ethylenediamine (0.04 ml_, d = 0.90 g / mL, 0.35 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 10b (15 mg, 45% yield). Subsequently 45 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.33 ml_), providing 10b (as the depicted HCI salt, 16 mg, 99% recovery) as a white solid.

[0399] 1H NMR (600 MHz, DMSO) 6 9.32 (s, 1H), 9.22 (t, J = 6.2 Hz, 1H), 8.92 (s, 1H), 8.33 (s, 1 H), 7.98 (br, 3H), 7.45 - 7.26 (m, 5H), 5.74 (s, 1 H), 3.64 (q, J = 6.2 Hz, 1 H), 3.05 (td, J = 6.2, 4.1 Hz, 1H).

[0400] 13C NMR (151 MHz, DMSO) 6 164.8, 148.4, 142.4, 141.6, 140.8, 139.8, 137.3, 136.2, 133.3, 130.7, 129.3, 128.9, 128.1, 127.3, 106.3, 48.0, 38.7, 37.0.

[0401] HRMS (ESI+) calculated for C22HI9CI3N5O+[M+H]+: 474.0650, found: 474.0653

[0402] 1-benzyl-A / -(2-(methylamino)ethyl)-4-(3,4,5-trichlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (10c)

[0403] Using General Procedure C above, methyl 1-benzyl-4-(3,4,5-trichlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 10a (30.0 mg, 0.07 mmol) and A / 1-methylethane- 1,2-diamine (0.04 mL, d = 0.85 g / mL, 0.35 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 10c (18 mg, 55% yield). Subsequently 18 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.38 mL), providing 10c (as the depicted HCI salt, 20 mg, 100% recovery) as a white solid.

[0404] 1H NMR (600 MHz, DMSO) 6 9.33 (s, 1H), 9.26 (t, J = 6.1 Hz, 1H), 8.93 (s, 1H), 8.66 (s, 1H), 8.33 (s, 1H), 7.52 - 7.25 (m, 5H), 5.74 (s, 2H), 3.68 (q, J = 6.1 Hz, 2H), 3.15 (p, J = 5.9 Hz, 2H), 2.59 (t, J = 5.9 Hz, 2H).

[0405] 13C NMR (151 MHz, DMSO) 0 164.9, 148.4, 142.3, 141.6, 140.7, 139.9, 137.3, 136.2, 133.2, 129.3, 128.9, 128.1, 127.3, 106.3, 48.3, 48.0, 35.7, 32.7. Proposed accidental equivalence for two aromatic carbons.

[0406] HRMS (ESI+) calculated for C23H2iCI3N5O+[M+H]+: 488.0806, found: 488.0810

[0407] methyl 1-benzyl-4-(3,4,5-trifluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (11a)

[0408] Using General Procedure B above, methyl 4-(3,4,5-trifluorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.3 mmol) and benzyl bromide (0.77 ml_, d = 1.44 g / mL, 5.6 mmol, 2 equiv) and cesium carbonate (1.59 g, 4.88 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 11a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (890 mg, 69%).[°409l IH NMR (400 MHz, CDCI3) 58.57 (dd, J= 9.4, 7.0 Hz, 2H), 8.20 (s, 1H), 8.17 (s, 1H), 7.43 - 7.36 (m, 3H), 7.26 - 7.18 (m, 2H), 5.46 (s, 2H), 4.04 (s, 3H).

[0410] 13C NMR (101 MHz DMSO) 6 165.2, 150.2 (ddd, J =1JCF= 246.2,2JCF = 9.9,3JCF = 3.7 Hz), 148.6, 142.6, 140.2, 139.9, 139.5 (dt,1JCF= 245.1 Hz,2JCF= 15.52 Hz) 139.3, 136.1, 133.3 (d,3JCF = 4.8 Hz), 128.9, 128.1, 127.4, 112.9 (d,2JCF = 22.4 Hz), 108.9, 52.5, 47.9.

[0411] 19F NMR (376 MHz, CDCI3) 0 -134.44 (d, J = 20.5 Hz, 2F), -158.99 (t, J= 20.5 Hz, 1F).

[0412] HRMS (ESI+) calculated for C2iHi5F3N3O2+[M+H]+: 398.1111, found: 398.1123

[0413] A / -(2-aminoethyl)-1-benzyl-4-(3,4,5-trifluorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (11b)

[0414] Using General Procedure C above, methyl 1-benzyl-4-(3,4,5-trifluorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 11a (30.0 mg, 0.07 mmol) and ethylenediamine (0.04 ml_, d = 0.90 g / mL, 0.35 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 11b (12 mg, 35% yield). Subsequently 12 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.28 ml_), providing 11b (as the depicted HCI salt, 13 mg, 99% recovery) as a white solid.

[0415] 1H NMR (600 MHz, DMSO) 6 9.22 (t, J = 6.1 Hz, 1H), 9.08 (dd, J = 10.0, 7.0 Hz, 2H), 8.90 (s, 1H), 8.31 (s, 1H), 8.01 (br, 3H), 7.48 - 7.21 (m, 5H), 5.73 (s, 2H), 3.64 (q, J = 6.1 Hz, 2H), 3.04 (q, J = 5.9 Hz, 2H).

[0416] 13C NMR (151 MHz, DMSO) 6 164.7, 150.3 (ddd,1JCF=244.9 Hz,2JCF = 9.2 Hz,3JCF = 3.2 Hz), 148.3, 142.2, 141.8, 140.8, 139.6, 139.5 (dt,1JCF=252.7 Hz,2JCF = 15.8 Hz), 136.2, 133.3, 128.9, 128.1, 127.3, 113.6 (dd,2JCF = 11.6 Hz,3JCF = 5.1 HZ), 105.9, 47.9, 38.7, 36.9.

[0417] 19F NMR (376 MHz, DMSO) 5 -135.02 (d,3JFF = 21.9 Hz, 2F), -159.90 (t,3JFF = 21.9 Hz, 1F).

[0418] HRMS (ESI+) calculated for C22HI9F3N5O+[M+H]+: 426.1536, found: 426.1543

[0419] methyl 1-benzyl-4-(3-bromo-5-chlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (12a)

[0420] Using General Procedure B above, methyl 4-(3-bromo-5-chlorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 2.7 mmol) and benzyl bromide (0.65 mL, d = 1.44 g / mL, 5.14 mmol, 2 equiv) and cesium carbonate (1.33 g,4.09 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 12a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (487 mg, 39%).

[0421] 1H NMR (400 MHz, DMSO) 5 8.93 (t, J = 1.6 Hz, 1H), 8.88 (s, 1H), 8.83 (t, J = 1.6 Hz, 1 H), 8.37 (s, 1 H), 7.80 (t, J = 1.6 Hz, 1 H), 7.41 - 7.26 (m, 5H), 5.72 (s, 2H), 3.93 (s, 3H).

[0422] 13C NMR (101 MHz, DMSO) 5 165.3, 148.6, 143.1, 140.2, 140.07, 139.43, 139.40, 136.1, 134.3, 131.4, 129.9, 128.9, 128.0, 127.44, 127.37, 122.3, 109.1, 52.5, 48.0.

[0423] HRMS (ESI+) calculated forC2iH16BrCIN302+[M+H]+: 456.0109, found: 456.0100

[0424] N-(2-aminoethyl)-1 -benzyl-4-(3-bromo-5-chlorophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (12b)

[0425] Using General Procedure C above, methyl 1-benzyl-4-(3-bromo-5-chlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 12a (50.0 mg, 0.11 mmol) and ethylenediamine (0.04 ml_, d = 0.90 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 12b (44 mg, 83% yield). Subsequently 44 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.9 mL), providing 12b (as the depicted HCI salt, 47 mg, 99% recovery) as a white solid.

[0426] 1H NMR (600 MHz, DMSO) 5 9.21 (t, J = 1.6 Hz, 1H), 9.16 (t, J = 6.2 Hz, 1H), 9.09 (t, J = 1.6 Hz, 1H), 8.89 (s, 1H), 8.31 (s, 1H), 7.86 (t, J = 1.6 Hz, 1H), 7.46 - 7.24 (m, 5H), 5.74 (s, 2H), 3.60 (q, J = 6.2 Hz, 2H), 3.00 (t, J = 6.2 Hz, 2H).

[0427] 13C NMR (151 MHz, DMSO) 5 164.7, 148.2, 142.5, 140.7, 140.1, 139.8, 136.3, 134.4, 131.6, 130.5, 128.9, 128.0, 127.9, 127.3, 122.4, 106.1, 47.9, 37.7. Two pairs of accidentally equivalent aromatic carbons, likely due to pseudosymmetry resulting from the electronic similarity of Cl and Br.

[0428] HRMS (ESI+) calculated for C22H2oBrCIN50+[M+H]+: 484.0534, found: 484.0541

[0429] 1-benzyl-4-(3-bromo-5-chlorophenyl)-N-(2-(methylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (12c)

[0430] Using General Procedure C above, methyl 1-benzyl-4-(3-bromo-5-chlorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 12a (50.0 mg, 0.11 mmol) and A / 1-methylethane- 1,2-diamine (0.04 mL, d = 0.85 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 12c (39 mg, 73% yield). Subsequently 39 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.8 mL), providing 12c (as the depicted HCI salt, 41 mg, 99% recovery) as a white solid.

[0431] 1H NMR (600 MHz, DMSO) 69.22 (t, J= 1.7 Hz, 1H), 9.20 (t, J= 6.2 Hz, 1H), 9.10 (d, J= 1.7 Hz, 1H), 8.90 (s, 1H), 8.31 (s, 1H), 7.86 (t, J = 1.7 Hz, 1H), 7.64 - 7.18 (m, 5H), 5.74 (s, 2H), 3.68 (q, J = 6.2 Hz, 2H), 3.13 (t, J = 6.2 Hz, 2H), 2.57 (s, 3H).

[0432] 13C NMR (151 MHz, DMSO) 5 164.9, 148.3, 142.5, 142.4, 140.7, 140.1, 139.8, 136.3, 134.4, 131.6, 130.5, 128.9, 128.1, 127.9, 127.3, 122.5, 106.1, 48.2, 47.9, 35.8, 32.7.

[0433] HRMS (ESI+) calculated for C23H22BrCIN5O+[M+H]+: 498.0691, found: 498.0695

[0434] 1-benzyl-4-(3-bromo-5-chlorophenyl)-N-(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (12d)

[0435] Using General Procedure C above, methyl 1-benzyl-4-(3-bromo-5-chlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 12a (50.0 mg, 0.11 mmol) and Af. AT-dimethylethane-1,2-diamine (0.06 ml_, d = 0.81 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 12d (45 mg, 84% yield). Subsequently 39 mg of thismaterial was converted to HCI salt using 0.1 M HCI in methanol (0.9 mL), providing 12d (as the depicted HCI salt, 48 mg, 99% recovery) as a white solid.

[0436] 1H NMR (600 MHz, DMSO) 59.98 (br, 1H), 9.21 (t, J= 1.6 Hz, 1 H), 9.19 (t, J = 6.2 Hz, 1 H), 9.09 (t, J = 1.6 Hz, 1 H), 8.90 (s, 1 H), 8.31 (s, 1 H), 7.87 (t, J = 1.6 Hz, 1 H), 7.42 - 7.23 (m, 5H), 5.74 (s, 2H), 3.71 (q, J = 6.2 Hz, 2H), 3.20 (br, 2H), 2.76 (s, 6H).

[0437] 13C NMR (151 MHz, DMSO) 6 164.7, 148.3, 142.5, 142.3, 140.7, 140.1, 139.9, 136.2, 134.4, 131.6, 130.4, 128.9, 128.1, 127.9, 127.3, 122.5, 106.1, 56.4, 48.0, 42.9, 34.9.

[0438] HRMS (ESI+) calculated for C24H24BrCIN5O+[M+H]+: 512.0847, found: 512.0856

[0439] methyl 1-benzyl-4-(4-bromophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (13a)

[0440] Using General Procedure B above, methyl 4-(4-bromophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.01 mmol) and benzyl bromide (0.72 mL, d = 1.44 g / mL, 6.02 mmol, 2 equiv) and cesium carbonate (1.47 g, 4.52 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 13a was further purified by silica gel column chromatography (0~40% Ethyl acetate in Hexane) and isolated as a white solid (1.1 g, 80%).

[0441] 1H NMR (400 MHz, DMSO) 58.86 (s, 1 H), 8.86 (d, J = 8.75 Hz, 2H), 8.36 (s, 1 H), 7.78 (d, J = 8.75 Hz, 2H), 7.40 - 7.28 (m, 5H), 5.72 (s, 2H), 3.92 (s, 3H).

[0442] 13C NMR (151 MHz, DMSO) 5 165.6, 148.2, 145.7, 140.1, 139.9, 139.5, 136.3, 135.9, 131.4, 130.9, 128.9, 128.0, 127.4, 123.3, 108.4, 52.4, 47.9.

[0443] HRMS (ESI+) calculated for C2iHi7BrN3O2+[M+H]+: 422.0499, found: 422.0484

[0444] N-(2-aminoethyl)-1 -benzyl-4-(4-bromophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (13b)

[0445] Using General Procedure C above, methyl 1-benzyl-4-(4-bromophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 13a (50.0 mg, 0.12 mmol) and ethylenediamine (0.06 mL, d = 0.90 g / mL, 0.59 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 13b (50 mg, 88% yield). Subsequently 50 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.1 mL), providing 13b (as the depicted HCI salt, 54 mg, 99% recovery) as a white solid.

[0446] 1H NMR (600 MHz, DMSO) 6 9.09 (t, J = 6.2 Hz, 1H), 8.97 (dd, J = 8.6, 1.6 Hz, 2H), 8.84 (s, 1H), 8.25 (s, 1H), 7.85 - 7.71 (br, 3H), 7.76 (dd, J = 8.6, 1.6 Hz, 2H), 7.40 - 7.26 (m, 5H), 5.72 (s, 2H), 3.62 (q, J = 6.2 Hz, 2H), 3.02 (t, J = 6.2 Hz, 2H).

[0447] 13C NMR (151 MHz, DMSO) 6 165.5, 148.3, 145.2, 142.8, 141.2, 140.2, 136.9, 136.4, 131.9, 131.8, 129.5, 128.6, 127.9, 123.9, 105.8, 48.5, 39.5, 37.8.

[0448] HRMS (ESI+) calculated for C22H2iBrN5O+[M+H]+: 450.0924, found: 450.0928

[0449] 1-benzyl-4-(4-bromophenyl)-N-(2-(methylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (13c)

[0450] Using General Procedure C above, methyl 1-benzyl-4-(4-bromophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate 13a (50.0 mg, 0.12 mmol) and / V1-methylethane- 1,2-diamine (0.06 mL, d = 0.85 g / mL, 0.59 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 13c (39 mg, 68% yield). Subsequently 39 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.86 mL), providing 13c (as the depicted HCI salt, 42 mg, 99% recovery) as a white solid.

[0451] 1H NMR (600 MHz, DMSO) 69.12 (t, J = 6.2 Hz, 1H), 8.99 (dd, J = 8.6, 1.6 Hz, 2H), 8.85 (s, 1H), 8.25 (s, 1H), 7.76 (dd, J = 8.6, 1.6 Hz, 2H), 7.46 - 7.23 (m, 5H), 5.72 (s, 2H), 3.66 (q, J = 6.2 Hz, 2H), 3.09 (t, J = 6.2 Hz, 2H), 2.55 (s, 3H).

[0452] 13C NMR (151 MHz, DMSO) 6 164.9, 147.8, 144.6, 142.2, 140.6, 139.6, 136.3, 135.8, 131.4, 131.2, 128.9, 128.0, 127.3, 123.3, 105.2, 48.4, 47.9, 35.9, 32.9.

[0453] HRMS (ESI+) calculated for C23H23BrN5O+[M+H]+: 464.1080, found: 464.1088

[0454] 1-benzyl-4-(4-bromophenyl)-A / -(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (13d)

[0455] Using General Procedure C above, methyl 1-benzyl-4-(4-bromophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 13a (50.0 mg, 0.12 mmol) and / V, A7-dimethylethane-1,2-diamine (0.06 mL, d = 0.81 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 13d (43 mg, 76% yield). Subsequently 43 mg of this material was converted to HOI salt using 0.1 M HCI in methanol (0.95 mL), providing 13d (as the depicted HCI salt, 46 mg, 99% recovery) as a white solid.

[0456] 1H NMR (600 MHz, DMSO) 59.73 (s, 1H), 9.13 (t, J = 6.1 Hz, 1H), 8.98 (dd, J = 8.6, 1.6 Hz, 2H), 8.85 (s, 1 H), 8.25 (s, 1 H), 7.77 (dd, J = 8.6, 1.6 Hz, 2H), 7.39 - 7.28 (m, 5H), 5.72 (br, 2H), 3.71 (q, J = 6.1 Hz, 2H), 3.25 (s, 2H), 2.79 (s, 6H).

[0457] 13C NMR (151 MHz, DMSO) 5 164.9, 147.8, 144.7, 142.1, 140.6, 139.6, 136.3, 135.8, 131.3, 131.3, 128.9, 128.0, 127.3, 123.4, 105.2, 56.4, 47.9, 42.8, 34.7.

[0458] HRMS (ESI+) calculated for C24H25BrN5O+[M+H]+: 478.1237, found: 478.1244

[0459] methyl 1-benzyl-4-(4-bromo-3-chlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (14a)

[0460] Using General Procedure B above, methyl 4-(4-bromo-3-chlorophenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 2.73 mmol) and benzyl bromide (0.65 mL, d = 1.44 g / mL, 5.46 mmol, 2 equiv) and cesium carbonate (1.33 g, 4.09 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 * 15 ml_). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 14a was further purified by silica gel column chromatography (0~40% Ethyl acetate in Hexane) and isolated as a white solid (775 mg, 62%).

[0461] 1H NMR (400 MHz, DMSO) 5 9.05 (d, J = 2.0 Hz, 1H), 8.90 (s, 1H), 8.69 (dd, J = 8.5, 2.0 Hz, 1H), 8.40 (s, 1H), 7.99 (d, J = 8.5 Hz, 1H), 7.40 - 7.26 (m, 6H), 5.73 (s, 2H), 3.93 (s, 3H).

[0462] 13C NMR (151 MHz, DMSO) 0 165.4, 148.5, 144.0, 140.2, 140.0, 139.5, 137.8, 136.2, 134.1, 133.3, 130.2, 128.9, 128.9, 128.1, 127.4, 122.8, 109.0, 52.5, 48.0.

[0463] HRMS (ESH-) calculated for C2iH16BrCIN3O2+[M+H]+: 456.0109, found: 456.0107

[0464] N-(2-aminoethyl)-1 -benzyl-4-(4-bromo-3-chlorophenyl)-1 H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (14b)

[0465] Using General Procedure C above, methyl 1-benzyl-4-(4-bromo-3-chlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 14a (50.0 mg, 0.11 mmol) and ethylenediamine (0.05mL, d = 0.90 g / mL, 0.59 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 14b (47 mg, 88% yield). Subsequently 47 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.95 mL), providing 14b (as the depicted HCI salt, 46 mg, 91% recovery) as a white solid.

[0466] 1H NMR (600 MHz, DMSO) 5 9.27 (d, J = 2.1 Hz, 1H), 9.14 (t, J = 6.2 Hz, 1H), 8.96 (dd, J = 8.5, 2.1 Hz, 1H), 8.88 (s, 1H), 8.29 (s, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.60 (br, 3H), 7.41 - 7.25 (m, 5H), 5.73 (s, 2H), 3.61 (q, J = 6.2 Hz, 2H), 3.01 (t, J = 6.3 Hz, 2H).

[0467] 13C NMR (151 MHz, DMSO) 6 164.8, 148.1, 143.1, 142.3, 140.7, 139.7, 137.7, 136.3, 133.8, 133.3, 130.6, 129.4, 128.9, 128.0, 127.3, 122.9, 105.8, 47.9, 37.4. Proposed accidental equivalence of two aromatic carbons.

[0468] HRMS (ESI+) calculated for C22H20BrCIN5O+[M+H]+: 484.0534, found: 484.0541

[0469] 1-benzyl-4-(4-bromo-3-chlorophenyl)-N-(2-(methylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (14c)

[0470] Using General Procedure C above, methyl 1-benzyl-4-(4-bromo-3-chlorophenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate (50.0 mg, 0.11 mmol) and / V-methylethane-1,2-diamine (0.06 mL, d = 0.85 g / mL, 0.59 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 14c (48 mg, 87% yield). Subsequently 48 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.1 mL), providing 14c (as the depicted HCI salt, 50 mg, 99% recovery) as a white solid.

[0471] 1H NMR (600 MHz, DMSO) 6 9.28 (d, J = 2.1 Hz, 1H), 9.18 (t, J = 6.2 Hz, 1H), 8.96 (dd, J = 8.5, 2.1 Hz, 1H), 8.89 (s, 1H), 8.47 (br, 2H), 8.29 (s, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.43 - 7.24 (m, 5H), 5.73 (s, 2H), 3.66 (q, J = 6.2 Hz, 3H), 3.12 (t, J = 6.2 Hz, 3H), 2.57 (s, 3H).

[0472] 13C NMR (151 MHz, DMSO) 5 164.9, 148.1, 143.1, 142.3, 140.7, 139.7, 137.7, 136.3, 133.8, 133.3, 130.6, 129.4, 128.9, 128.0, 127.3, 122.9, 105.8, 48.4, 47.9, 35.8, 32.9.

[0473] HRMS (ESI+) calculated for C23H22BrClN5O+[M+H]+: 498.0691, found: 498.0705

[0474] 1-benzyl-4-(4-bromo-3-chlorophenyl)-N-(2-(dimethylamino)ethyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (14d)

[0475] Using General Procedure C above, methyl 1-benzyl-4-(4-bromo-3-chlorophenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 14a (50.0 mg, 0.11 mmol) and Af. / V-dimethylethane-1,2-diamine (0.06 ml_, d = 0.81 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 14d (37 mg, 67% yield). Subsequently 37 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.8 mL), providing 14d (as the depicted HCI salt, 40 mg, 99% recovery) as a white solid.

[0476] 1H NMR (600 MHz, DMSO) 6 9.27 (d, J = 2.1 Hz, 1H), 9.19 (t, J = 6.1 Hz, 1H), 8.96 (dd, J = 8.5, 2.1 Hz, 1H), 8.89 (s, 1H), 8.29 (s, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.41 - 7.27 (m, 5H), 5.73 (s, 2H), 3.71 (q, J = 6.1 Hz, 3H), 3.23 (br, 2H), 2.78 (s, 6H).

[0477] 13C NMR (151 MHz, DMSO) 6 164.7, 148.1, 143.1, 142.2, 140.7, 139.7, 137.7, 136.3, 133.8, 133.3, 130.6, 129.4, 128.9, 128.0, 127.3, 122.9, 105.8, 56.4, 47.9, 42.8, 34.8.

[0478] HRMS (ESI+) calculated for C24H24BrClN5O+[M+H]+: 512.0847, found: 512.0851

[0479] methyl 1-benzyl-4-(4-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (15a)

[0480] Using General Procedure B above, methyl 4-(4-(trifluoromethyl)phenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.11 mmol) and benzyl bromide (0.74 mL, d = 1.44 g / mL, 6.23 mmol, 2 equiv) and cesium carbonate (1.52 g, 4.67 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 * 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 15a was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) and isolated as a white solid (537 mg, 42%).

[0481] 1H NMR (400 MHz, DMSO) 6 8.97 (d, J= 8.2 Hz, 1H), 8.90 (s, 1H), 8.41 (s, 1H), 7.95 (d, J = 8.2 Hz, 2H), 5.73 (s, 2H), 3.93 (s, 3H).

[0482] 13C NMR (151 MHz, DMSO) 5 165.5, 150.8 (q,1JCF= 272.93 Hz), 148.6, 145.2, 140.6, 140.4, 140.2, 139.7, 136.2, 129.6, 129.4 (q,2JCF = 30.75 Hz), 128.9, 128.1, 127.4, 125.3 (d,3JCF= 3.8 Hz), 108.9, 52.5, 47.9.

[0483] 19F NMR (376 MHz, DMSO) 5 -61.03.

[0484] HRMS (ESI+) calculated for C22H17F3N3O2+[M+H]+: 412.1267, found: 412.1278

[0485] N-(2-aminoethyl)-1-benzyl-4-(4-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (15b)

[0486] Using General Procedure C above, methyl 1-benzyl-4-(4-(trifluoromethyl)phenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 15a (50.0 mg, 0.12 mmol) and ethylenediamine (0.04 mL, d = 0.90 g / mL, 0.61 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 15b (47 mg, 85% yield). Subsequently 47 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (1.1 mL), providing 15b (as the depicted HCI salt, 50 mg, 99% recovery) as a white solid.

[0487] 1H NMR (400 MHz, MeOD) 69.20 (s, 1H), 8.69 (d, J= 8.2 Hz, 2H), 8.42 (s, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.51 - 7.27 (m, 5H), 5.76 (s, 2H), 3.79 (t, J = 5.4 Hz, 2H), 3.24 (t, J = 5.4 Hz, 2H).

[0488] 13C NMR (151 MHz, MeOD) 6 167.2, 148.5, 147.1, 144.7, 141.6, 140.6, 136.9, 135.8, 132.8 (q,2JCF = 32.1 Hz), 131.3, 130.4, 129.9, 129.0, 126.6 (q,3JCF= 4.0 Hz), 125.6 (q,1JCF= 273.4 Hz), 107.5, 51.1, 41.1, 38.6.

[0489] 19F NMR (376 MHz, MeOD) 5 -64.22.

[0490] HRMS (ESI+) calculated for C23H20F3N5O+[M+H]+: 440.1693, found: 440.1698.

[0491] 1-benzyl-N-(2-(methylamino)ethyl)-4-(4-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (15c)

[0492] Using General Procedure C above, methyl 1-benzyl-4-(4-(trifluoromethyl)phenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate 15a (50.0 mg, 0.12 mmol) and A / 1-methylethane- 1,2-diamine (0.06 mL, d = 0.85 g / mL, 0.59 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 15c (38 mg, 69% yield). Subsequently 38 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.86 mL), providing 15c (as the depicted HCI salt, 40 mg, 97% recovery) as a white solid.

[0493] 1H NMR (400 MHz, MeOD) 59.19 (s, 1H), 8.71 (d, J= 8.2 Hz, 2H), 8.43 (s, 1H), 7.91 (d, J = 8.2 Hz, 2H), 7.46 - 7.33 (m, 5H), 5.76 (s, 2H), 3.82 (t, J = 5.6 Hz, 2H), 3.29 (br, 2H)(overlapped with MeOD), 2.75 (s, 3H).

[0494] 13C NMR (151 MHz, MeOD) 6 167.4, 148.5, 147.1, 144.7, 141.6, 140.6, 137.0, 135.8, 132.8 (d,2JCF = 32.3 Hz), 131.3, 130.4, 129.9, 128.9, 126.4 (q,1JCF= 254.63 Hz), 126.6 (q,3JCF= 3.9 Hz), 107.5, 51.1, 50.8, 37.4, 33.9.

[0495] 19F NMR (376 MHz, MeOD) 5 -64.24.

[0496] HRMS (ESI+) calculated for C24H23F3N5O+[M+H]+: 454.1849, found: 454.1850

[0497] 1-benzyl-N-(2-(dimethylamino)ethyl)-4-(4-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (15d)

[0498] Using General Procedure C above, methyl 1-benzyl-4-(4-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate 15a (50.0 mg, 0.12 mmol) and / V -dimethylethane-1,2-diamine (0.06 ml_, d = 0.81 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 15d (46 mg, 80% yield). Subsequently 46 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.86 ml_), providing 15d (as the depicted HCI salt, 49 mg, 98% recovery) as a white solid.

[0499] 1H NMR (600 MHz, MeOD) 69.50 (s, 1H), 8.57 (d, J= 8.3 Hz, 2H), 8.51 (s, 1H), 7.94 (d, J = 8.3 Hz, 2H), 7.54 - 7.34 (m, 5H), 5.83 (s, 2H), 3.90 (t, J = 5.8 Hz, 2H), 3.47 (t, J = 5.8 Hz, 2H), 3.00 (s, 6H).

[0500] 13C NMR (151 MHz, MeOD) 0 166.7, 148.3, 146.9, 145.3, 141.4, 139.9, 135.2, 134.7, 132.9 (q,2JCF= 32.4 Hz), 131.3, 130.4, 130.1, 129.2, 126.8 (q,3JCF = 3.9 Hz), 125.5 (q,1JCF= 271.3 Hz), 107.9, 58.7, 51.6, 43.9, 36.2.

[0501] 19F NMR (376 MHz, MeOD) 6 -64.27.

[0502] HRMS (ESI+) calculated for C25H25F3N5O+[M+H]+: 468.2006, found: 468.2012

[0503] methyl 1-benzyl-4-(3-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxylate (16a)

[0504] Using General Procedure B above, methyl 4-(3-(trifluoromethyl)phenyl)-1 / 7-imidazo[4,5-c]pyridine-6-carboxylate (synthesized by general procedure A) (1g, 3.11 mmol) and benzyl bromide (0.74 mL, d = 1.44 g / mL, 6.23 mmol, 2 equiv) and cesium carbonate (1.52 g, 4.67 mmol, 1.5 equiv) were combined and heated to 60 °C for 24 h. TLC at this point indicated complete consumption of the starting material. The reaction mixture was extracted with EtOAc (3 * 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The product 16a was further purified by silica gel column chromatography (0~40% Ethyl acetate in Hexane) and isolated as a white solid (665 mg, 42%).

[0505] 1H NMR (400 MHz, DMSO) 69.18 (s, 1H), 9.05 (d, J = 7.79 Hz, 1H), 8.89 (s, 1H), 8.39 (s, 1 H), 7.89 - 7.76 (m, 2H), 7.37 - 7.29 (m, 5H), 5.73 (s, 2H), 3.93 (s, 3H).

[0506] 13C NMR (101 MHz, DMSO) 5 165.4, 148.4, 144.9, 140.14, 140.10, 139.6, 137.7, 136.2, 132.6, 129.4, 128.9, 128.0, 127.4, 125.9, 125.3 (q,3JCF = 4.5 Hz), 124.5 (q,1JCF= 271.4 Hz), 108.7, 52.4, 47.9. We propose two aromatic carbons are accidentally equivalent.

[0507] 19F NMR (376 MHz, DMSO) 5 -61.10.

[0508] HRMS (ESI+) calculated for C22H17F3N3O2+[M+H]+: 412.1267, found: 412.1277

[0509] 1-benzyl-N-(2-(dimethylamino)ethyl)-4-(3-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]pyridine-6-carboxamide hydrochloride (16d)

[0510] Using General Procedure C above, methyl 1-benzyl-4-(4-(trifluoromethyl)phenyl)-1 / - / -imidazo[4,5-c]pyridine-6-carboxylate 15a (50.0 mg, 0.12 mmol) and Af. AT-dimethylethane-1,2-diamine (0.04 mL, d = 0.81 g / mL, 0.55 mmol, 5 equiv) were combined and heated to 90 °C for 20 h, affording the free base of 16d (42 mg, 78% yield). Subsequently 46 mg of this material was converted to HCI salt using 0.1 M HCI in methanol (0.95 mL), providing 16d (as the depicted HCI salt, 45 mg, 98% recovery) as a white solid.

[0511] 1H NMR (600 MHz, MeOD) 69.11 (d, J = 2.2 Hz, 1H), 8.88 (s, 1H), 8.77 (d, J= 7.8 Hz, 1H), 8.42 (d, J= 2.2 Hz, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.81 (t, J = 7.8 Hz, 1H), 7.46 - 7.32 (m, 5H), 5.75 (d, J = 2.2 Hz, 2H), 3.89 (t, J = 5.7, Hz, 2H), 3.45 (d, J = 5.7 Hz, 2H), 3.00 (s, 6H).

[0512] 13C NMR (151 MHz, MeOD) 5 164.6, 145.8, 144.48, 141.5, 138.9, 135.2, 135.0, 133.1, 129.2 (d,2JCF= 32.3 Hz), 127.8, 127.5, 127.0, 126.1, 124.9 (3JCF= 3.8 Hz), 124.6 (3JCF= 3.8 Hz), 122.8 (q,1JCF = 272.1 Hz), 104.6, 56.0, 48.1, 41.1, 33.4. Proposed accidental equivalence of two aromatic carbons.

[0513] 19F NMR (376 MHz, DMSO) 5 -69.91.

[0514] HRMS (ESI+) calculated for C25H25F3N5O+[M+H]+: 468.2006, found: 468.2019

[0515] Synthesis of Purine Analogs

[0516] Exemplary purine analogs were prepared by using described methods below and depicted in FIG. 3.

[0517] Synthesis of P-1

[0518] To a 1 L flask vial equipped with magnetic stir bar containing 2,6-dichloropurine (40.0 g), 3,4-Dihydropyran (2.0 equiv), para-Toluenesulfonic acid (1 mol%), ethyl acetate (400 mL) was added. The reaction flask was capped and heated at 80 °C for 1 h. TLC typically indicated complete consumption of the 2,6-dichloropurine, the reaction was cooling to room temperature, the reaction mixture was extracted with aq. NaHCO3(1 M, 3X 100 mL) to remove the para-Toluenesulfonic acid. The organic fractions were combined and concentrated in vacuo to afford in P-1 (53 g, 90%) as a white solid. This material was used in the next step without further purification.

[0519] Synthesis of P-3a

[0520] To an argon-flushed 20 mL vial equipped with magnetic stir bar were added 2,6-dichloro-9-(tetrahydro-2 / - / -pyran-2-yl)-9 / - / -purine P-1 (1.0g), the 2-(3,4,5-trichlorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.6 equiv), and sodium carbonate (2.0 equiv) in Dioxane (20 mL), and water (2 mL). The reaction mixture was purged with argon for 30 minutes. Subsequently, Pd(PPh3)4(5 mol%) was added quickly under a stream of argon, and the reaction vial was sealed and heated at 100 °C for 20 hours. After completion, the reaction was cooled to room temperature and was extracted with EtOAc (3 * 15 mL). The combined organic phase were washed with brine, dried with sodium sulfate, and concentrated in vacuo. The crude product was further purified by silica gel column chromatography (0~40% EtOAc in Hexane) afford in P-3a (1.3 g, 85%) as a white solid.

[0521] Synthesis of P-3b

[0522] To a 50 mL flask with magnetic stir bar containing 2-chloro-9-(tetrahydro-2 / - / -pyran-2-yl)-6-(3,4,5-trichlorophenyl)-9 / - / -purine P-3a (1.0 g), tetrabutylammonium bromide (0.4 equiv), cesium fluoride (2.0 equiv), 18-crown-6 (0.4 equiv) was added in acetonitrile (15 mL). The reaction flask was capped and heated at 80 °C for 12 h. The reaction was cooling to room temperature, the reaction mixture was extracted with water to remove all water-soluble impurities. The organic fractions were combined and concentrated in vacuo to afford the crude product as a light-yellow solid. 19F NMR was used to prove the exchange of the chlorine to fluorine. Crude P-3b was used in the next step without further purification.

[0523] Synthesis of P-3c

[0524] To an argon-flushed 20 mL vial equipped with magnetic stir bar were added 2-fluoro-9-(tetrahydro-2H-pyran-2-yl)-6-(3,4,5-trichlorophenyl)-9H-purine P-3b (1.0 g), potassium cyanide (2.0 equiv), and 1,4-diazabicyclo[2.2.2]octane (20 mol%) in DMSO (10 mL), and water (10 mL). The reaction vial was sealed and heated at 100 °C for 12 hours. After completion, the reaction was cooled to room temperature and was extracted with EtOAc (3 x 15 mL). The combined organic phase were washed with brine, dried with sodium sulfate, and concentrated in vacuo. The crude product was further purified by silica gel column chromatography (0~40% EtOAc in Hexane) to give P-3c as a white solid (560 mg, 37% over 2 steps). Note: all glassware were rinsed with sodium chlorite solution to quench excess cyanide residual before cleaning.

[0525] Synthesis of P-3d

[0526] To a 50 mL flask equipped with magnetic stir bar containing 9-(tetrahydro-2H-pyran-2-yl)-6-(3,4,5-trichlorophenyl)-9 / 7-purine-2-carbonitrile P-3c (1.0 g), TFA (5 mL) was added in DCM (5 mL). The reaction flask was capped and stirred at room temperature for 12 h. The reaction mixture was extracted with aq. NaHCOs (1 M, 3 X 30 mL). The organic fractions were combined and concentrated in vacuo to afford crude product P-3d as a light-yellow solid. It was used in the next step without further purification.

[0527] Synthesis of P-3d1

[0528] To a 20 mL vial equipped with magnetic stir bar containing 6-(3,4,5-trichlorophenyl)-9 / - / -purine-2-carbonitrile P-3d (200mg) and cesium carbonate (1.5 equiv), benzyl bromide (1.2 equiv) was added dropwise in DMF at room temperature. The vial was capped and stirring at 80°C for 16h, the reaction was cooled to room temperature. TLC typically indicated complete consumption of the oxidation intermediate. The reaction mixture was extracted with EtOAc (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate ormagnesium sulfate, and concentrated in vacuo. The benzylation product was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) affording P-3d1 as a white solid (270 mg, 89% over 2 steps).

[0529] Synthesis of P-3e1

[0530] To a 10 mL microwave reaction tube equipped with magnetic stir bar containing 9-benzyl-6-(3,4,5-trichlorophenyl)-9H-purine-2-carbonitrile P-3d1 (100mg) in anhydrous methanol, thionyl chloride (2.0 equiv) was added dropwise in at 0°C. The vial was capped and stirring in the microwave reactor at 60°C for 5h, After completion, the reaction was cooled to room temperature. The reaction mixture was quenched in aq. NaHCO3. The reaction mixture was extracted with EtOAc (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The benzylation product was further purified by silica gel column chromatography (0-40% Ethyl acetate in Hexane) affording P-3e1 as a white solid (98 mg, 60%).

[0531] Synthesis of P-3f1

[0532] To a 1-dram vial equipped with magnetic stir bar were added methyl 9-benzyl-6-(3,4,5-trichlorophenyl)-9H-purine-2-carboxylate P-3e1 (30 mg) and a magnetic stir bar was added in anhydrous toluene. The vial was sealed with a rubber septum and purged with N2 for 5 minutes. Ethylene diamine (10 equiv) was added via syringe, and 10 mol% Ca(OH)₂ was added as amidation catalyst, the mixture was stirred at rt for 5 minutes, and then heated to 90 °C for 2-30 h, until TLC indicated complete consumption of the starting material. The reaction was allowed to cool to room temperature, then extracted with DCM (3 x 15 mL). The combined organic phases were washed with brine, dried with sodium sulfate or magnesium sulfate, and concentrated in vacuo. The free base of amide was further purified by silica gel column chromatography (0-20% Methanol in Dichloromethane) as a white solid. A portion of this material was dissolved in 0.1 M HCI in methanol (prepared from 12 M HCI (aq.)) and concentrated in vacuo. The salt was again dissolved in methanol (-1 -4 mL) and concentrated in vacuo to remove residual water overnight to afford P-3f1 as the HCI salt (12 mg, 35%).

[0533] 1H NMR (600 MHz, MeOD) 0 9.12 (s, 1H), 8.68 (s, 1H), 7.49 - 7.45 (m, 2H), 7.39 -7.30 (m, 3H), 5.67 (s, 2H), 3.84 (t, J= 5.9 Hz, 2H), 3.28 (t, J = 5.9 Hz, 2H).

[0534] 13C NMR (151 MHz, MeOD) 5 166.4, 154.8, 152.1, 150.8, 149.9, 137.2, 136.5, 135.5, 134.9, 132.9, 131.1, 130.1, 129.6, 129.2, 48.4, 39.0.

[0535] HRMS (ESI) calcd for C19H14Cl3N6O [M+H]+: 475.0602, found: 475.0608.

[0536] Biological Evaluation

[0537] Plasmodium falciparum cultures. The P. falciparum strain Dd2 (MRA-150), 3D7 (MRA-102) and NF54 (MRA-1000) were obtained from MR4 (ATCC, Manassas, VA; BEI Resources, NIAID, NIH). The P. falciparum NF54-PfK13-C580Y line was kindly supplied by David Fidock17. The P. falciparum Pikine-Pfcoronin-R100K-E107V mutant line and the parental Pikine strain were kindly supplied by Dyann Wirth18. All P. falciparum strains were maintained in O+positive human erythrocytes (Grifols, Memphis, TN, USA) at 5% hematocrit in RPMI 1640 media (Thermo Fisher Scientific, Waltham, MA) containing 2 g / L glucose, 5.94 g / L HEPES, 2.3 g / L sodium bicarbonate, 5 g / L Albumax I, and 50 mg / L hypoxanthine. All reagents were obtained from Sigma-Aldrich (St. Louis, MO). Media was supplemented with 20 mg / L of gentamicin (Thermo Fisher Scientific, Waltham, MA). Parasite cultures were maintained at 37°C under reduced oxygen conditions (5% CO2, 5% O2, and 90% N2) with shaking. Highly synchronous ring stage cultures (>98%) were obtained through two consecutive 5% sorbitol treatments (Sigma-Aldrich, St. Louis, MO) performed 6 hours apart.

[0538] In vitro selection of drug resistance. Resistance selection was assessed using a single-step method2023. For the single step selection, inoculums of 1 x 109synchronous parasites from newly cloned P. falciparum 3D7 strain or Dd2-Pol6 line were subjected to continuous drug pressure at three times the EC90values. Each condition was performed in three independent cultures. For the first 7 days, media containing drug was replaced daily, then every other day until day 14. A parallel control culture was maintained under identical conditions with an equal volume of DMSO. After 14 days, drug pressure was removed, and cultures were monitored daily by Giemsa-stained smears with media changes every other day for 60 days. Fresh blood (50 pL) was added weekly to sustain culture viability.

[0539] P. falciparum morphological assessments. Ring stage NF54-WT strain parasite cultures (>98%) were synchronized by two consecutive 5% sorbitol treatment (Sigma-Aldrich, St. Louis, MO), administered 6 hours apart, 48 hours prior to the initiation of the experiment. All cultures were maintained at 37°C under reduced oxygen conditions (5% CO2, 5% O2, and 90% N2) with continuous shaking. To evaluate morphological alterations following drug treatment, compounds were applied at specific concentrations and durations. Thin blood smears were prepared at indicated time points, fixed with 100% methanol (Sigma-Aldrich, St. Louis, MO, USA), and stained for 15 minutes using a 20% Giemsa solution (Sigma-Aldrich; diluted in deionized water). Following smear preparation, parasites were immediately gassed and returned to 37°C with shaking. Morphological growth phenotypes were assessed in biological duplicates.

[0540] HepG2 and liver stage assays. HepG2 cells (Homo sapiens hepatoblastoma, ATCC) were maintained in collagen-coated T-75 flasks with sugar-free DMEM (Gibco), supplementedwith 10% FBS (Corning), 25 mM glucose (Millipore-Sigma), 1 mM sodium pyruvate (Corning), 5 pg / mL penicillin, 5 pg / mL streptomycin, 10 pg / mL neomycin, and 2 mM L-glutamine (Gibco). Cultures were incubated at 37°C in a humidified atmosphere containing 5% CO2. Cells were detached using TrypLE (Gibco) once cultures reached 60-90% confluence. Cell density was determined via trypan blue exclusion with a hemocytometer. Subsequently, 17,500 cells per well were dispensed into collagen-coated 384-well plates (Greiner Bio-One) using a Biomek NXPautomated workstation (Beckman Coulter), 24 hours prior to sporozoite infection.

[0541] Luciferase-expressing P. berghei ANKA strain GFP-Lucama1-eef1a (line 1052cl 1) was obtained from the Sporocore at the University of Georgia, as previously described24. Sporozoites were isolated according to established protocols, utilizing bicarbonate-free RPMI (KD Medical) as the collection buffer25. HepG2 cells were infected with 2,000 sporozoites per well. At three hours post-infection, 40 nL of each compound dilution was transferred from dose-response source plates to assay plates using a pin tool (V& P Scientific) mounted on a Biomek NXP, achieving a final test concentration of 1x in media. The plates were incubated for 44 hours at 37°C in a humidified atmosphere containing 5% CO2. After incubation, plates were fixed with 4% paraformaldehyde (Thermo Scientific) in PBS for 20 minutes. Following fixation, the plates were washed twice by adding and removing 20 pL of PBS per well. Plates were stained with 50 ng / mL mouse monoclonal antibody 13.3 (anti-GAPDH) sourced from The European Malaria Reagent Repository (http: / / www.malariaresearch.eu) diluted in a permeabilization and blocking buffer containing 0.3% Triton X and 1% BSA, and incubated overnight at 4°C. After three PBS washes, plates were treated with 2 pg / mL goat anti-mouse AlexaFluor 488 (Invitrogen), diluted in stain buffer, and incubated overnight at 4°C. Subsequently, three PBS washes were performed before counterstaining with 10 pg / mL Hoechst 33342 (Invitrogen) at room temperature for 30 minutes, followed by two additional washes. Imaging was conducted using an ImageXpress Micro Confocal high content system (Molecular Devices). Schizont number and area were normalized using positive (MMV390048) and negative (DMSO) controls, and hepatocyte nuclei counts were used as an indicator of toxicity.

[0542] P. falciparum Growth Inhibition Assay. Dose-dependent growth inhibition with the reported compounds were evaluated using a 10-point dilution series and the in vitro SYBR Green I assay as a readout. Synchronous ring-stage parasites (1% starting parasitemia, 1% hematocrit) were cultured in 96-well half-area dark plates and continuously exposed to each compound for 72 h at 37 °C under reduced oxygen conditions (5% CO2, 5% O2, and 90% N2). After 72 h, parasite growth was assessed by the SYBR Green I assay as previously described26. SYBR Green I was excited at 485 nm and its emission was measured at 535 nmusing a Cytation5 plate reader (Agilent BioTek). Parasite growth was normalized to untreated control parasites and calculated as a percentage. Background was determined using uninfected red blood cells (RBCs). Dose-dependent assays were performed in at least two biological replicates and two technical replicates. Reported values represent the mean of biological replicates with standard error of the mean (SEM). Initial concentrations were optimized after compound screening to ensure the EC50fell within the tested range. DMSO concentrations were maintained <0.02% in all assays. Data were fitted using a four-parameter logistic dose-response curve, and half-maximal effective concentration (EC50) values were calculated using GraphPad Prism (GraphPad Software, Inc.). Assays comparing EC50values across different parasite lines were performed concurrently.

[0543] Dose-dependent Asexual Blood Stage Susceptibility Assays. Assays were performed as previously described8with slight modifications20. Parasites of the P. falciparum 3D7 strain were synchronized in the ring stage 48 h prior to the assay by two rounds of 5% sorbitol treatment (Sigma-Aldrich, St. Louis, MO) with a 6 h interval between treatments. After parasites completed one lifecycle and reinvaded RBCs, an additional sorbitol treatment was performed on the day of the assay to obtain highly synchronous early rings. Synchronized infected RBCs were plated in five 96-well plates and sequentially exposed to compounds for 8-hour windows starting at the following developmental stages: early rings (0-8 h), late rings (8-16 h), early trophozoites (16-24 h), late trophozoites (24-32 h), and schizonts (32-40 h). After each exposure window, compounds were removed by three rounds of washing with prewarmed RPMI to avoid growth delays, and infected RBCs were transferred to a new plate. Plates were incubated under standard culture conditions, and parasite growth was assessed 72 h after the start of the assay using the SYBR Green I assay as described above. A 72-hour continuous exposure dose-response curve was included in parallel as control. Dose-response data were fitted using a four-parameter logistic dose-response curve or, where appropriate, a biphasic dose-response, and EC50values were calculated using GraphPad Prism (GraphPad Software, Inc.). Assays were performed in at least two independent biological replicates, each with two to four technical replicates.

[0544] A similar experimental design was used to determine the EC50values in ring stages following 6- or 8-hour exposure periods in a dose-dependent manner using different strains and mutants. Assays comparing EC50values across different parasite lines were performed concurrently. These assays were performed in one or two independent biological replicates, each with two to four technical replicates.

[0545] Modified Recrudescence and DHA-induced Dormancy Survival Assays. Assays were performed as previously described.23Recrudescence was monitored in bulk cultures(FIG. 6B-7B). Parasites were synchronized twice with 5% sorbitol, with a 6 h interval between treatments. Thirty-six hours after the final synchronization, 5 mL cultures of early ring stages (0-6 h post-invasion) were adjusted to 3% parasitemia and 5% hematocrit and treated with DHA (700 nM) or PRC1910 (500 nM) for 6 h at 37°C with shaking under reduced oxygen conditions (5% CO2, 5% O2, and 90% N2). Following incubation, parasites were washed three times with pre-warmed complete RPMI media and transferred to new flasks containing drug-free media. Thin blood smears were prepared every 24 h over a 30 day period, fixed with 100% methanol (Sigma-Aldrich, St. Louis, MO, USA), and stained with 20% Giemsa for 15 min (Sigma-Aldrich; diluted in deionized water). After smear preparation, cultures were immediately gassed and returned to 37°C with shaking. Media was replaced every other day, and 50 pL of fresh blood was added weekly. Recrudescence was assessed by light microscopy, scoring the proportion of viable parasites with normal morphology following drug removal. For each condition, 10,000 erythrocytes were independently evaluated by two microscopists under blinded conditions, and results were reported as percentages.

[0546] The effect of PRC analogs on DHA-induced dormant parasites was assessed as previously described.23Bulk cultures were synchronized and treated with 700 nM DHA for 6 h as described above (FIG.7B). At 24 h post-treatment, parasites were treated with 250 nM and 500 nM of PRC1910 or DMSO (mock) for 6 h. Final DMSO concentrations did not exceed 0.02%. After incubation, parasites were washed three times with pre-warmed complete RPMI media and transferred to new flasks containing drug-free media. Recrudescence was monitored by light microscopy as described above for up to 30 days.

[0547] RESULTS AND DISCUSSION

[0548] P. falciparum ABS growth inhibition potencies of imidazopyridine and purine analogs

[0549] Referring to Table 1, numerous imidazopyridine analogs were evaluated with respect to the growth of P. falciparum ABS.Table 1. P. falciparum Dd2 strain growth inhibition potencies of imidazo[4,5-c]pyridine carboxamide antimalariais (Bn = benzyl; Ph = phenyl).

[0550] Based on preliminary testing, imidazopyridine analogs with specific substitution patterns were evaluated. Referring to Table 2, imidazopyridine analogs 3b-3g were all poor growth inhibitors, with EC50values ranging from 1,000-5,000 nM. Hypothesizing that the poor potency was due to lower lipophilicity (cf. 1 and 3c), N1-alkylation of 3a was carried out,affording 4a-6a (FIG. 1). Note that the regioselectivity of alkylation was expected based on steric effects and was confirmed by NOE studies of 6a. These esters were then amidated as described for 3a. Progression along the series 3b (A / 1-H), 4b (A / 1-Me), 5b (A / 1-Bu), and 6n (A / 1-Bn) is instructive, confirming the importance of lipophilicity for growth inhibition (Table 2). Compounds 3b (A / 1-H) and 4b ( / V1-Me) both have very poor potency, but relative to 3b, a 10-fold improvement is seen for 5b ( / V1-Bu), and a 25-fold improvement is seen for 6b (A / 1-Bn, EC50 = 209 ± 6 nM). Methylation and dimethylation of the pendant basic N did not have much effect on growth inhibition potency (cf. 6b, 6c, 6d). Dimethylation of the ethylene chain proximal to the amide proved deleterious (cf. 6b, 6e), but dimethylation distal to the amide caused a smaller increase in EC50 (cf. 6b, 6f). Lastly, we prepared butyl amide 6g; as expected, its poor growth inhibition potency (EC50> 2,000 nM) demonstrates the need for a basic nitrogen in the amide substituent, mimicking the SAR seen for 1 and 2.Table 2. P. falciparum (Dd2 strain, 72 h, SYBR Green) ABS growth inhibition potencies of imidazo[4,5-c]pyridine analogs®aAII compounds were tested as HCI salts; growth inhibition (EC50 values) of ABS was determined using SYBR Green I assay at 72 h endpoint. Values represent average ± S. E. M from at least two biological replicates (with two technical replicates). P. falciparum Dd2 strain is multi-drug resistant. For compounds that did not achieve 100% growth inhibition at 10,000 nM, EC50 values are reported as “> X” where X is the highest concentration at which no inhibition was observed.

[0551] Next, a study of the effect of substitution on the 4-aryl ring was conducted, which corresponds to the 1-aryl ring of 1 and 2. Ten different substitution patterns were examined, 6 of which imparted good potency in the p-carboline series. For simplicity we focused on amides of ethylenediamines (various methylation levels, see e.g., FIG. 2). ABS growth inhibition EC50values for these 25 compounds are shown in Table 3, where the value for 6b is repeated for reference.

[0552] As can be seen, 3',4'-difluoro (7b-d) and 4'-chloro-3'-fluoro (8b-d) analogs are less potent than 3',4'-d ichloro analog 6b. A slight improvement was seen for 3'-chloro-4'-fluoro (9b-d) analogs relative to 4'-chloro-3'-fluoro (8b-d). But the best potencies were seen for 3',4',5'-trichloro analogs 10b-10c. Compound 10b (EC50= 43 ± 2) is roughly 4-fold more potent than 6b. This SAR trend mimics what we previously observed in the p-carboline series.

[0553] Seeking lower molecular weight, we examined 3',4',5'-trifluoro analog 11b, which was more potent than, 3',4'-dichloro analog 6b, but 3-fold less potent than 3',4',5'-trichloro 10b.Compounds 12b-d, featuring the 3'-bromo,5'-chlorophenyl substitution pattern proved favorable for potency, just as it had been seen in the p-carboline series. However compounds 13b-d featuring 4'-bromophenyl substitution had poor growth inhibition potency. Good potency was rescued by addition of a 3'-chloro group: 4'-bromo,3'-chlorophenyl analogs 14b-d are slightly more potent than 3',4'-dichloro analogs 6b-d. Although 4'-trifluoromethylphenyl analogs proved potent in the p-carboline series, the corresponding imidazo[4,5-c]pyridine analogs 15b-d were not potent. Lastly, 4'-trifluoromethylphenyl analogs were not potent in the P-carboline series, and 4'-trifluoromethylphenyl imidazo[4,5-c]pyridine 16d was also a very poor growth inhibitor.Table 3. P. falciparum (Dd2 strain, 72 h, SYBR Green) ABS growth inhibition potencies of benzylated imidazo[4,5-c]pyridine analogs3aAII compounds were tested as HCI salts; growth inhibition (EC50values) of ABS was determined using SYBR Green I assay at 72 h endpoint. Values represent average ± S. E. M from at least two biological replicates (with two technical replicates). P. falciparum Dd2 strain is multi-drug resistant. For compounds that did not achieve 100% growth inhibition at 10,000 nM, EC50values are reported as “> X” where X is the highest concentration at which no inhibition was observed.

[0554] Finally, a study of purine analogs was conducted. Fifteen different substitution patterns were examined, 6 of which imparted good potency in the p-carboline series. ABS growth inhibition EC5o values for these 15 compounds are shown in Table 4.Table 4. P. falciparum (Dd2 strain, 72 h, SYBR Green) ABS growth inhibition potencies of purine analogs3aAII compounds were tested as HCI salts; growth inhibition (EC50values) of ABS was determined using SYBR Green I assay at 72 h endpoint. Values represent average ± S. E. M from at least two biological replicates (with two technical replicates). P. falciparum Dd2 strain is multi-drug resistant. For compounds that did not achieve 100% growth inhibition at 10,000 nM, EC50values are reported as “> X” where X is the highest concentration at which no inhibition was observed.

[0555] In vitro ADME-tox studies

[0556] Four imidazopyridine analogs were submitted for ADME-Tox profiling: 6b, 6c, 6d, and 10b (Table 5). As can be seen, the most potent compound 10b has acceptable buffer solubility (8.5 pM), moderate log D (3.62 at pH 7.4), > 3 hour half-life in both human liver microsomes and rat hepatocytes, and > 100-fold selectivity both for cytotoxicity to HepG2 cells and hERG inhibition.Table 5. Physicochemical properties, metabolic stability, HepG2 cytotoxicity and hERG block of 1a and selected analogs

[0557] Compounds are refractory to resistance selection in vitro

[0558] Imidazopyridine 6d (PRC1824) and purine P-2h1 (PRC1868) were selected. A single step of resistance selection was performed by subjecting flasks containing an inoculum of 1 x109P. falciparum parasites (drug-sensitive 3D7 strain) to 6d and purine P-2h1 at a concentration of three times the EC90 value for 14 days. In parallel, a flask treated with an equal volume of DMSO was used as a reference control. After removing drug pressure, cultures were monitored for 60 days. In both treatments, parasite clearance occurred within the first four days, and no parasite recrudescence was observed after 60 days. From these experiments, the minimum inoculum of resistance (MIR) was determined to be > 9 (i.e., >109infected red blood cells).

[0559] Compounds that do not produce resistant parasites from large inoculum (~109asexual blood stage parasites) are called “irresistible” or “resistance-refractory” and are valuable for drug development due to their high resistance barrier. T 0 distinguish true resistance-refractory compounds from those limited by low genetic diversity, we used the Dd2-Polδ line with defective DNA polymerase δ proofreading, which increases mutation rates under drug pressure, to assess if 10b and P-3f1 were also “irresistible”.

[0560] After removing drug pressure, cultures were monitored for 60 days. In both treatments, parasite clearance occurred within the first four days, and no parasite recrudescence was observed after 60 days. From these experiments, the minimum inoculum of resistance (MIR) for 10b and P-3f1 was also determined to be > 9 (i.e., >109infected red blood cells). Thus, compounds 6d, P-2h1, 10b and P-3f1, and presumably other members of the imidazo[4,5-c]pyridine and purine classes, have a high barrier to resistance.

[0561] To assess potential cross-resistance / sensitivity within this scaffold, 10b and P-3f1 were evaluated using the Antimalarial Resistome Barcode Sequencing (AReBar) pooled-screening assay.6This methodology employs genetically barcoded Plasmodium falciparum parasite lines, which were previously developed using CRISPR / Cas9 genome editing to introduce a barcode cassette into a nonessential locus of each line. Barcode sequencing subsequently quantifies the relative abundance of each line. Using this strategy, 51 distinct cell lines are pooled for direct competition assays in the presence of selected compounds, thereby encompassing diverse resistance mechanisms within both the 3D7 and Dd2 genetic backgrounds. These mutations cover a range of different biological functions including protein synthesis, nucleic acid synthesis and processing, mitochondrial function, and hemoglobin uptake and processing, thus addressing potential mechanisms of antimalarial action.

[0562] Growth inhibition EC50values were measured against the drug-resistant Dd2 strain and susceptible 3D7 strain P. falciparum, using a similar 72 h SYBR Green protocol. The resistome pool was exposed to 0.18 pM of 10b and 0.27 pM of P-3f1 for 14 days. Cultures were run in triplicate in 1000 pL and the parasitemia was maintained between 0.5-5%.Cumulative growth profiles for drug treated and no drug controls were prepared. Positive control treatments yield expected expansions in CARL (GNF179) and DHODH (DSM265) mutants. Drug pressure was maintained at the indicated concentrations throughout the assay for 10b and P-3f1, and the pool exhibited no growth recovery for either compound. A barcode profile was recovered for analysis of the pool. Differential analysis using the R package DESeq2 was performed. Cell lines with an average barcode base count of less than 50 were filtered out. Treated samples with growth are compared to day 14 controls, and those with suppressed growth to day 4 controls. A significant expansion is defined as a cell exceeding a LFC (treatment vs. control) of 2.5 and a percentage population of 3%. There were no mutant cell lines that past these thresholds in 10b and P-3f1 treated samples, thus, no crossresistance was detected.

[0563] Morphological observations of Giemsa-stained smears by light microscopy

[0564] Under normal conditions, parasites proceed through the intraerythrocytic asexual blood stage cycle within 36 to 48 h, from new infection of merozoites, to the ring, to trophozoite, to schizont stage which burst the red blood cell, releasing the merozoites to infect new red cells.

[0565] Ring-stage parasites (Dd2 strain), synchronized by double sorbitol treatment, were exposed to 6d, P-2h1, 10b, P-3f1, DHA or chloroquine continuously for 72 hours. Within 24 hours of treatment, they collapsed into a pyknotic or dormant-like form and remained there for the 72-hour observation period, similar to those treated with 700 nM DHA. In contrast, parasites treated with chloroquine advanced to trophozoite stage before becoming pyknotic at 72 hours.

[0566] Dose-dependent susceptibility profile during the intraerythrocytic asexual cycle and liver stages ofP. berghei

[0567] The potential of 10b in inhibiting liver stage development of Plasmodium berghei was assessed. No inhibition was seen at concentrations up to 5 pM. The activity observed at 10 pM was due to toxicity to HepG2 cells observed at 10 pM.

[0568] To determine if this novel class targets specific stages within the asexual blood cycle, we performed the asexual blood stage susceptibility assay. This assay also enables the identification of potential mechanisms of action for new antimalarial drugs by comparing their susceptibility profiles to those of established antimalarials.7 8Parasites of the P. falciparum 3D7 strain were synchronized in the ring stage 48 h prior to the assay by two rounds of 5% sorbitol treatment with a 6 h interval between treatments. After parasites completed one lifecycle and reinvaded red blood cells (RBCs), an additional sorbitol treatment was performedon the day of the assay to obtain highly synchronous early rings. Synchronized infected RBCs were plated in five 96-well plates and sequentially exposed to compounds for 8-hour windows starting at the following developmental stages: early rings (0-8 h), late rings (8-16 h), early trophozoites (16-24 h), late trophozoites (24-32 h), and schizonts (32-40 h) (FIG. 4). After each exposure period, compounds were removed by three rounds of washing with pre-warmed RPMI to avoid growth delays, and infected RBCs were transferred to a new plate. Plates were incubated under standard culture conditions, and parasite growth was assessed 72 h after the start of the assay using the SYBR Green I assay. A 72-hour continuous exposure doseresponse curve was included in parallel as control. Dose-response data were fitted using a four-parameter logistic dose-response curve, and EC5o values were calculated using GraphPad Prism (GraphPad Software, Inc.). Assays were performed in at least two independent biological replicates, each with two to four technical replicates.

[0569] In the blood stage susceptibility assays, since compounds are removed after each exposure period and parasites continue to grow without them until the assay ends, the stagespecific assay can measure how effectively a compound kill parasites at each stage. The EC508 h values represent the stage-specific half maximal lethal doses, while the EC5072 h values, determined from assays where parasites are continuously exposed to compounds, reflect the cytostatic (growth-inhibiting) effects of those compounds. As illustrated in FIG. 4, 10b exhibited a stage-specific profile similar to DHA, showing lethal potency across all stages with EC50 values comparable to those observed after 72 hours of continuous drug exposure. Compounds 6d and 6b were most potent against parasites during early trophozoite stages, but their potency was still within two-fold of that observed with 72-hour continuous exposure in other stages. These observations suggest that 10b rapidly kills ring-stage P. falciparum.

[0570] Parasite Reduction Ratio Assay

[0571] Optimal pharmacodynamic characteristics for a candidate drug include rapid onset of action and rapid parasite elimination. Accurate assessment of these parameters requires distinguishing between viable and nonviable parasites — a process complicated by the potential for viable parasites to exhibit metabolic inactivity, while dying parasites may remain metabolically active and morphologically unchanged. The in vitro parasite reduction ratio (PRR) assay quantifies viable parasites and yields critical pharmacodynamic metrics, such as PRR, 99.9% parasite clearance time (PCT99.9%), and lag phase.9

[0572] As shown in FIG. 5, compounds 10b (mustard line, filled diamonds) and purine P-3f1 (gray line, open circles) rapidly reduce the number of viable parasites, faster than chloroquine (dashed magenta line, filled circles) or pyrimethamine (brick red line, upward triangles), and nearly as fast as artesunate (dashed blue line, squares). Within 24 h, 10b and P-3f1 eachreduced the parasite count by more than 3 log units, with 99.9% parasite clearance times of (PCT99.9%) of 24 and 21.5 h respectively.

[0573] PRR represents the reduction in parasitemia over one 48 h asexual reproduction cycle.15Since the starting log parasitemia was only 5, and there were no viable parasites left at 48 h, the log PRR will be > 5. However, a close look at the plot indicates a steeper slope from 0 to 24 h than from 24 h to 48 h. Thus, based on the slope from 0 to 24 h, the extrapolated log PRR values for 10b and P-3f1 are both >8. This puts them in the category of artesunate, which also has a log PRR >8. In contrast, the log PRR of compound 1 (data not shown) is only 2.79. Thus, 10b and P-3f1 have favorable quick-killing profiles that 1 does not share.

[0574] Activity against proliferating ring stages: extended ring stage recrudescence assay

[0575] Effective antimalarials must clear ring-stage parasites before they become dormant or develop into transmissible gametocytes. The stage specificity assay indicates that P-3f1 is halting the development of ring stages (both early and late) similarly to DHA. To determine whether parasites exhibiting dormant-like morphology observed in 10b-treated cultures could also recrudesce beyond the 72 hours tested in the stage specificity assay, we performed ring recrudescence assays following 6 h of exposures to 10b (FIGS 6A-6C). A concentration of 500 nM of 10b in NF54-WT (DHA-sensitive) was selected corresponding to 0% growth in the ring stage dose-dependent 6-hour exposure curve (FIG. 6A). As expected, after a 6-hour treatment with 700 nM DHA followed by removal of treatment, recrudescence occurred after 3 days (FIG. 6B). In contrast, cultures treated with 500 nM 10b showed no recrudescence within 30 days post-treatment. These findings indicate that a 6-hour exposure to 10b efficiently eliminates ring-stage P. falciparum parasites, thereby preventing recrudescence beyond the 72-h period assessed in the ring stage dose-dependent curve.

[0576] Activity against DHA-induced dormant parasites as potential combination therapy

[0577] There is increasing evidence suggesting that activity against artemisinin derivative-induced dormant rings should be considered the "gold standard" for evaluating potential partner drugs.11"14This property could enable the future development of novel ACTs even in the context of artemisinin resistance (ART-R) as well as the advancement of innovative new combinations. To assess whether 10b display activity against DHA-induced dormant parasites, we treated highly synchronous rings cultures (6-8 h post-invasion) with 700 nM DHA for 6 h. Following incubation, drug was removed and incubated in drug-free media for 18 h to ensure that parasites displayed morphological dormancy, at which time these parasites weretreated with either 10b at 250 nM and 500 nM, or DMSO for only 6 hours (FIGS. 7A-7B). Following an initial 6-hour exposure to 700 nM DHA (DHA / mock), recrudescence occurred after 3 days similar to the ring survival assay. Remarkably, when DHA-induced dormant parasites were subsequently treated with 10b, no proliferating parasites were detected for up to 30 days at both concentrations. These findings indicate that 10b effectively eliminate DHA-induced dormant parasites.

[0578] Effectiveness against parasites that resist DHA due to Kelch 13-C580Y and Pikine-Pfcoronin-R100K-E107V mutations

[0579] To better evaluate the susceptibility of the ring-stage to 10b, the stage susceptibility assay was performed, which compared the P. falciparum NF54 wildtype strain and NF54-K13-C580Y line, expressing K13 wildtype (WT) orC580Y mutation, respectively17. PRC1584 was more potent against the PfK13-C580Y mutant while PRC1910 was slightly more potent. As expected, DHA was slightly less potent against the NF54-K13-C580Y line compared to the NF54-WT. We also evaluated PRC 1584 and 10b against parasites carrying Pfcoronin mutations (Pikine-Pfcoronin-R100K-E107V), which also appear to mediate resistance to DHA18. The Pikine-Pfcoronin-R100K-E107V mutant line exhibited similar susceptibility to PRC 1584 and 10b as the parental Pikine strain after 8 hours of exposure of exposure. These data further support the potency of these compounds against artemisinin resistant parasites. The EC50values are summarized in Table 6.Table 6. EC5o values used for the bar graphs.

[0580] In vivo pharmacokinetic studies

[0581] Pharmacokinetic analysis of 10b in male CD1 mice was performed by Pharmaron (Beijing, China). Three mice each (ranging from 22.1 -23.6 g) were used for oral (40 mg / kg,gavage, conscious) and for IV (10 mg / kg, lateral tail vein with anesthesia) administration. The IV vehicle was 10% DMSO, 90% (20% (w / v) HP-p-CD in water); the PO vehicle was 20% DMSO, 80% (20% (w / v) HP-p-CD in water). Drug was dosed at 2 mg / mL IV, 4 mg / mL PO (clear solutions). Quantitation was performed by triple-quad LC-MS. Concentration vs time data and key pharmacokinetic parameters are shown in Tables 7-10.Table 7. IV (10 mg / kg dose) concentration vs time dataData in bold were used to calculate terminal elimination t1 / 2.Table 8. PO (40 mg / kg) concentration vs time dataData in bold were used to calculate terminal elimination t1 / 2.Table 9. IV pharmacokinetic parameters (10 mg / kg)Table 10. PO (40 mg / kg) pharmacokinetic parameters

[0582] In vivo pharmacodynamic studies

[0583] Efficacy of 10b in Plasmodium yoelii-infected mice

[0584] 10b was assessed for in vivo efficacy in a murine model of malaria (Plasmodium yoelii). In this modified 4-day Peters test19, mice were inoculated with parasites from a donor mouse (day 0) and then dosed orally with 10b on each of the subsequent 4 days (days 1-4) at the indicated doses or as a single dose on day 1 (see table in FIG. 8A). On day 5 of the experiment, the parasitemia for each mouse was determined microscopically by examining methanol-fixed and Giemsa-stained blood smears.

[0585] Mice were considered cured of infection if no parasites were detected in the blood 30 days following initial drug administration. 10b demonstrated good tolerability in mice, with no observable signs of toxicity such as changes in weight, fur condition, or grooming behavior. The compound achieved complete cure in all treated mice (4 / 4) at a single dose of 160 mg / kg, as well as at a regimen of 60 mg / kg / day. In contrast, chloroquine (CQ) at 60 mg / kg administered over four days was not effective in achieving cure in mice. Experiments were repeated with doses of 0.362, 2.5, 10 and 60 mg / kg / day to obtain the ED50 and ED90 doses and parasitemia for each mouse was determined microscopically by examining methanol-fixed and Giemsa-stained blood smears at day 5 (FIG. 8B). The ED50 and ED90values were assessed by generating dose-response curves using GraphPad Prism. The procedures involved, together with all matters relating to the care, handling, and housing of the animals used in this study, were approved by the Portland VA Medical Center Institutional Animal Care and Use Committee.

[0586] It should be emphasized that the above-described embodiments of the present disclosure are merely possible examples of implementations set forth for a clear understanding of the principles of the disclosure. Many variations and modifications may be made to the above-described embodiment(s) without departing substantially from the spirit and principles of the disclosure. All such modifications and variations are intended to be included herein within the scope of this disclosure and protected by the following claims.REFERENCES(1) World Health Organization. World malaria report 2024: addressing inequity in the global malaria response.; 2024.(2) Teuscher, F.; Gatton, M. L.; Chen, N.; Peters, J.; Kyle, D. E.; Cheng, Q. Artemisinin-induced dormancy in plasmodium falciparum: duration, recovery rates, and implications in treatment failure. J Infect Dis 2010, 202 (9), 1362-1368. DOI: 10.1086 / 656476 From NLM Medline.(3) Witkowski, B.; Lelievre, J.; Barragan, M. J.; Laurent, V.; Su, X. Z.; Berry, A.; Benoit-Vical, F. Increased tolerance to artemisinin in Plasmodium falciparum is mediated by a quiescence mechanism. Antimicrob Agents Chemother 2010, 54 (5), 1872-1877. DOI: 10.1128 / aac.01636-09 From NLM.(4) Dembele, L.; Gupta, D. K.; Lim, M. Y.; Ang, X.; Selva, J. J.; Chotivanich, K.; Nguon, C.; Dondorp, A. M.; Bonamy, G. M. C.; Diagana, T. T.; et al. Imidazolopiperazines Kill both Rings and Dormant Rings in Wild-Type and K13 Artemisinin-Resistant Plasmodium falciparum In Vitro. Antimicrob Agents Chemother 2018, 62 (5). DOI: 10.1128 / AAC.02235-17 From NLM Medline.(5) Kreutzfeld, O.; Orena, S.; Okitwi, M.; Tumwebaze, P. K.; Byaruhanga, O.; Katairo, T.; Conrad, M. D.; Legac, J.; Garg, S.; Crudale, R.; et al. Ex vivo susceptibilities to ganaplacide and diversity in potential resistance mediators in Ugandan Plasmodium falciparum isolates. Antimicrob Agents Chemother 2024, 68 (9), e0046624. DOI: 10.1128 / aac.00466-24 From NLM Medline.(6) Carrasquilla, M.; Drammeh, N. 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Insights into the intracellular localization, protein associations and artemisinin resistance properties of Plasmodium falciparum K13. PLoS Pathog 2020, 16 (4), e1008482. DOI: 10.1371 / journal.ppat.1008482 From NLM Medline.(18) Demas, A. R.; Sharma, A. I.; Wong, W.; Early, A. M.; Redmond, S.; Bopp, S.; Neafsey, D. E.; Volkman, S. K.; Hartl, D. L.; Wirth, D. F. Mutations in Plasmodium falciparum actin-binding protein coronin confer reduced artemisinin susceptibility. Proc Natl Acad Sci U S A 2018, 115 (50), 12799-12804. DOI: 10.1073 / pnas.1812317115 From NLM Medline.(19) Peters, W.; Fleck, S. L.; Robinson, B. L.; Stewart, L. B.; Jefford, C. W. The chemotherapy of rodent malaria. LX. The importance of formulation in evaluating the blood schizontocidal activity of some endoperoxide antimalarials. Ann Trop Med Parasitol 2002, 96 (6), 559-573. DOI: 10.1179 / 000349802125001744 From NLM Medline.(20) Bremers, E. K.; Butler, J. H.; Do Amaral, L. S.; Merino, E. 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Claims

CLAIMS1. A compound of formula Ior a pharmaceutically acceptable salt thereof,whereinrepresents a single or double bond;Z is CH or N;L1 is substituted or unsubstituted alkylene;l_2is substituted or unsubstituted alkylene group or substituted or unsubstituted cycloalkylene;Ri is H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocycloalkyl;R3and R4are independently H, alkyl, or hydroxyalkyl, or R3and R4together with the N atom to which they are attached form a heterocycloalkyl; andYi, Y2, and Y3are independently selected from the group consisting of H, halide, and haloalkyl.

2. The compound of claim 1, wherein Z is CH.

3. The compound of claim 1, wherein Z is N.

4. The compound of any one of claims 1-3, wherein Ri is substituted or unsubstituted phenyl.

5. The compound of claim 1, wherein Li is -(CH2)-.

6. The compound of claim 1, wherein R2is H.

7. The compound of claim 1, wherein L2is -(CH2)2-.

8. The compound of claim 1, wherein R3and R4are each H.

9. The compound of claim 1, wherein R3is H and R is alkyl.

10. The compound of claim 1, wherein11. The compound of claim 1, wherein Yi, Y2 and Y3 are independently Cl or F.

12. The compound of claim 1, wherein13. The compound of claim 1, wherein the compound has the formula IIIIor the pharmaceutically acceptable salt thereof,whereinZ is CH or N;R1 is substituted or unsubstituted phenyl;R3 and R4 are independently H or alkyl; andY1, Y2, and Y3 are independently selected from the group consisting of H and halide, wherein at least two of Y1, Y2, and Y3 are halide.

14. The compound of claim 13, wherein Z is CH.

15. The compound of claim 13, wherein Z is N.

16. The compound of claim 13, wherein R1 is unsubstituted phenyl.

17. The compound of claim 13, wherein R3and R4are each H.

18. The compound of claim 13, wherein Y1, Y2, and Y3are each chloro.of claim 13, wherein20. The compound of claim 13, wherein the compound has the formula III or IVor the pharmaceutically acceptable salt thereof.

21. A pharmaceutical composition comprising the compound of any one of claims 1 -20 and a pharmaceutically acceptable carrier.

22. A method for treating or preventing malaria in a subject, the method comprising administering to the subject an effective amount of the compound of any one of claims 1-20.

23. The method of claim 22, wherein the subject is infected with a strain of Plasmodium spp.

24. The method of claim 23, wherein the species of the genus Plasmodium comprises Plasmodium falciparum, Plasmodium vivax, Plasmodium knowlesi, Plasmodium ovalecurtisi, Plasmodium ovalewallikeri, or Plasmodium malariae.

25. The method of claim 22, wherein the method treats one or more symptoms of malaria.

26. The method of claim 25, wherein the symptom comprises swollen lymph nodes, fever, moderate-to-severe shaking chills, profuse sweating, diarrhea, anemia, cough, muscle and join aches, fatigue, headache, rapid breathing, rapid heart rate, sore throat, nausea and vomiting, enlarged spleen, mild jaundice, enlargement of the liver, brain inflammation and swelling (cerebral malaria), seizures, confusion or impairedconsciousness, vision problems, coma, circulatory collapse / shock, pulmonary edema or acute respiratory distress syndrome (ARDS), acidosis, acute kidney injury, abnormal bleeding or disseminated intravascular coagulation (DIC).

27. The method of claim 22, wherein after administration of the compound, a parasite that infected the subject with malaria is no longer present in the blood of the subject after 30 days of administration of the compound to the subject.

28. The method of claim 22, wherein the compound is administered orally, intravenously, or intramuscularly to the subject.

29. A method for treating or preventing an infection by Trypanosoma cruzi or Naegleria fowleri in a subject, the method comprising administering to the subject an effective amount of the compound of any one of claims 1-20.

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