Treatment of PKU with a corrector of mammalian SLC6A19 function

Abstract

Disclosed are compounds, compositions, and methods useful for treating or preventing diseases or disorders associated with abnormal amino acid concentrations through modulation of SLC6A19 transport.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 406,443, filed September 14, 2022. [Background technology]

[0002] Phenyleketonuria (PKU) is an inborn error of metabolism caused by mutations in phenylalanine hydroxylase (PAH), an enzyme responsible for the metabolism of phenylalanine. PKU is an autosomal recessive metabolic disorder in which phenylalanine is not properly metabolized, resulting in abnormally high plasma levels of phenylalanine. Individuals with PKU have abnormally high blood levels of phenylalanine. If untreated, this can lead to irreversible neurological damage and various complications, including intellectual disability, seizures, and neurodevelopmental and behavioral disorders. PKU is difficult to treat because blood levels of phenylalanine are directly related to diet. Patients must adhere to a strict lifelong diet, which impacts every aspect of their lives. The current standard of care is enzyme cofactor therapy and enzyme replacement therapy, but these therapies are not effective in all patients and carry potential risks of adverse events.

[0003] The enzyme responsible for metabolizing phenylalanine and thus maintaining phenylalanine homeostasis is phenylalanine hydroxylase (PAH). Loss-of-function (LOF) mutations in the PAH gene on chromosome 12q23.2 are known to cause most forms of PKU. These LOF mutations that cause PKU can be diagnosed as classic PKU (the most severe form) and less severe forms of "mild PKU" or "hyperphenylalaninemia." In addition to PAH, mutations in other enzymes that affect phenylalanine metabolism, such as dihydropteridine reductase (DHPR), an enzyme involved in the synthesis of cofactors required for PAH activity, can also increase phenylalanine concentrations. In addition to diet, blood amino acid concentrations, including phenylalanine concentrations, are regulated by SLC6A19, which is located in the proximal tubule of the kidney and is responsible for reabsorption of amino acids and returning them to the blood. Summary of the Invention

[0004] One aspect of the present invention provides compounds, compositions, and methods useful for treating or preventing diseases or disorders associated with abnormal amino acid concentrations through modulation of SLC6A19 transport.

[0005] Accordingly, provided herein are compounds having the structure of formula (I): [ka] (In the formula, n is 0 or 1, L1 is absent or selected from -NH-, -N(CH3)-, -O-, and -CH2-; L2 is -alkyl-; L3 is -(5-membered heteroaryl)-; X1 is -C(R1)(R2)(R3), X2 is optionally substituted aryl or heteroaryl; X3 is selected from -H, alkyl, and haloalkyl; R1 is selected from -H, halo, hydroxyl, amido, amino, alkylamino, and aminoalkyl; R2 and R3 are each independently selected from -H and alkyl, or R2 and R3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl; with the proviso that the compound is not selected from: [ka] Or a pharmaceutically acceptable salt thereof.

[0006] Another aspect of the present invention relates to a method of treating or preventing a disease or disorder associated with a genetic deficiency in phenylalanine hydroxylase in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0007] Another aspect of the present invention relates to a method of treating or preventing phenylketonuria, hyperphenylalaninemia, tyrosinemia, nonketotic hyperglycinemia, isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, a urea cycle disorder, or hyperammonemia in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0008] Another aspect of the present invention relates to a method of modulating SLC6A19 transport in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0009] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. Additionally, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0010] Other features, objects, and advantages of the invention will become apparent from the detailed description and claims. [Brief explanation of the drawings]

[0011] [Figure 1] 1 is a table summarizing isoleucine transport data for exemplary compounds of the invention: A = IC50<500 nM; B = IC50 500 nM-1500 nM; C = IC50>1500 nM-5000 nM; D = IC50>5000 nM-10000 nM; and E = IC50>10000 nM. DETAILED DESCRIPTION OF THE INVENTION

[0012] definition For convenience, before further description of the present invention, certain terms employed in the specification, examples, and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and should be understood by one of ordinary skill in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0013] In order to more readily understand the present invention, certain terms and phrases are defined below and throughout the specification.

[0014] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0015] The term "and / or," as used in the specification and claims, should be understood to refer to "either or both" of the elements so combined, i.e., elements that are conjunctive in some cases and disjunctive in other cases. Elements listed with "and / or" must be arranged in the same manner, i.e., "one or more of," the conjunctive elements. Other elements, whether related or unrelated to those elements specifically identified, may optionally be present other than the elements specifically identified in the "and / or" clause. Thus, as a non-limiting example, when used in conjunction with open-ended language such as "comprising," a reference to "A and / or B" can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); or in yet another embodiment, to both A and B (optionally including other elements).

[0016] As used herein in the specification and claims, "or" shall be understood to have the same meaning as "and / or," as defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as inclusive, i.e., including not only at least one, but also two or more of a number or list of elements, and optionally including additional items not listed. Only clearly indicated terms, such as "only one of," or "exactly one of," or, when used in the claims, "consisting of," shall mean the inclusion of exactly one element of a number or series of elements. Generally, as used herein, the term "or" shall only be interpreted as indicating exclusive alternatives (i.e., "one or the other, but not both") when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of." When used in the claims, "consisting essentially of" shall have its ordinary meaning as used in the field of patent law.

[0017] As used herein in the specification and claims, the phrase "at least one" in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not excluding any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently, "at least one of A and / or B") can refer in one embodiment to at least one, optionally, two or more As, and no B (and, optionally, including elements other than B); in another embodiment to at least one, optionally, two or more Bs, and no A (and, optionally, including elements other than A); in yet another embodiment to at least one, optionally, two or more As, and at least one, optionally, two or more Bs (and, optionally, including other elements); and so forth.

[0018] It should also be understood that, unless expressly indicated otherwise, in any method claimed herein that includes more than one step or action, the order of the method steps or actions is not necessarily limited to the order in which the method steps or actions are recited.

[0019] In the claims and the above specification, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like, shall be understood to be open-ended, i.e., to mean including, but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as defined in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

[0020] Certain compounds contained in the compositions of the present invention may exist in particular geometric forms or stereoisomers. In addition, the polymers of the present invention may also be optically active. The present invention contemplates that all such compounds are within the scope of the present invention, including cis- and trans-isomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are intended to be included in the present invention.

[0021] "Geometric isomer" refers to isomers that differ in the orientation of substituent atoms relative to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. Atoms (other than H) on each side of a carbon-carbon double bond can be in the E configuration (substituents on opposite sides of the carbon-carbon double bond) or the Z configuration (substituents on the same side). "R," "S," "S*," "R*," "E," "Z," "cis," and "trans" refer to structures relative to the core molecule. Certain disclosed compounds can exist in "atropisomeric" forms or as "atropisomers." Atropisomers are stereoisomers resulting from hindrance of rotation about a single bond, where the steric strain hindrance to rotation is sufficiently high to allow for separation of conformers. The compounds of the present invention can be prepared as individual isomers either by isomer-specific synthesis or by resolution from a mixture of isomers. Classical resolution techniques include using an optically active acid to form a salt of the free base of each isomer of the isomeric pair (followed by fractional crystallization and regeneration of the free base), using an optically active amine to form a salt of the acid form of each isomer of the isomeric pair (followed by fractional crystallization and regeneration of the free acid), using an optically pure acid, amine, or alcohol to form an ester or amide of each isomer of the isomer pair (followed by chromatographic separation and removal of the chiral auxiliary), or resolving the isomeric mixture of either the starting materials or the final product using a variety of well-known chromatographic methods.

[0022] For example, if a particular enantiomer of a compound of the present invention is desired, that enantiomer can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, separating the resulting mixture of diastereomers, and cleaving the auxiliary to obtain the desired pure enantiomer. Alternatively, if the molecule contains a basic functional group such as amino, or an acidic functional group such as carboxyl, diastereomeric salts can be formed with an appropriate optically active acid or base, followed by separation of the diastereomers so formed by fractional crystallization or chromatographic methods well known in the art, followed by recovery of the pure enantiomers.

[0023] A mole fraction purity percentage is the mole ratio of an enantiomer (or diastereomer), or the ratio of moles of an enantiomer (or diastereomer) to moles of its optical isomer. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least about 60%, about 70%, about 80%, about 90%, about 99%, or about 99.9% pure by mole fraction relative to other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least about 60%, about 70%, about 80%, about 90%, about 99%, or about 99.9% pure by mole fraction. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least about 60%, about 70%, about 80%, about 90%, about 99%, or about 99.9% pure by mole fraction.

[0024] When a disclosed compound is named or depicted by structure without indicating stereochemistry, and the compound has at least one chiral center, the name or structure should be understood to encompass any enantiomer of the compound, free of the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating stereochemistry, and the compound has two or more chiral centers, the name or structure should be understood to encompass a diastereomer free of the other diastereomer, multiple diastereomers free of other diastereomeric pairs, a mixture of diastereomers, a mixture of diastereomeric pairs, a mixture of diastereomers enriched in one diastereomer relative to the other diastereomer(s), or a mixture of diastereomers enriched in one or more diastereomers relative to the other diastereomers. The present invention encompasses all of these forms.

[0025] Structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, hydrogen replaced with deuterium or tritium, or carbon replaced with 13 C or 14 Compounds produced by substituting C-enriched carbons are within the scope of the present invention.

[0026] The term "prodrug," as used herein, encompasses compounds that are converted into therapeutically active agents under physiological conditions. A common method for making a prodrug is to include selected moieties that hydrolyze under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by enzymatic activity in the host animal.

[0027] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, that is involved in carrying or transporting the subject chemical entity from one organ or body part to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, not injurious to the patient, and substantially non-pyrogenic. Some examples of materials that can function as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and These include soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer; and (21) other non-toxic, compatible substances employed in pharmaceutical formulations. In certain embodiments, the pharmaceutical compositions of the present invention are non-pyrogenic, i.e., do not cause a significant temperature increase when administered to a patient.

[0028] The term "pharmaceutically acceptable salts" refers to relatively non-toxic inorganic and organic acid addition salts of a compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in free base form with a suitable organic or inorganic acid and isolating the salt so formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and lauryl sulfate. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts," J. Pharm. Sci. 66:1-19.)

[0029] In other cases, compounds useful in the methods of the present invention may contain one or more acidic functional groups, thereby enabling them to form pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these instances, the term "pharmaceutically acceptable salts" refers to the relatively non-toxic, inorganic and organic base addition salts of the compound(s). These salts can also be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in their free acid form with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, ammonia, or a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, e.g., Berge et al., supra).

[0030] The term "pharmaceutically acceptable cocrystal" refers to a solid coformer that does not form formal ionic interactions with the small molecule.

[0031] A "therapeutically effective amount" (or "effective amount") of a compound, with respect to use in treatment, means the quantity of compound(s) in a preparation that, when administered as part of a desired dosing regimen (to a mammal, preferably a human), reduces the symptoms, ameliorates the condition, or delays the onset of a disease or condition, e.g., in accordance with clinically acceptable standards for treating a disease or condition or for cosmetic purposes, at a reasonable benefit / risk ratio applicable to any medical treatment.

[0032] The term "prophylactic or therapeutic" treatment is art-recognized and includes administration to a host of one or more of the subject compositions. If the treatment is administered prior to the appearance of clinical symptoms of an undesired condition (e.g., a disease or other undesired condition in the host animal), the treatment is prophylactic (i.e., protects the host from the development of the undesired condition), whereas if the treatment is administered after the appearance of the undesired condition, the treatment is therapeutic (i.e., aims to reduce, ameliorate, or stabilize an existing undesired condition or its side effects).

[0033] The term "patient" or "subject" refers to a mammal in need of a particular treatment. In certain embodiments, the patient is a primate, dog, cat, or horse. In certain embodiments, the patient is human.

[0034] Aliphatic chains include the classes alkyl, alkenyl, and alkynyl, as defined below. Straight aliphatic chains are limited to unbranched carbon chain moieties. As used herein, the term "aliphatic group" refers to a straight, branched, or cyclic aliphatic hydrocarbon group, and includes saturated and unsaturated aliphatic groups, such as alkyl, alkenyl, or alkynyl groups.

[0035] "Alkyl" refers to a fully saturated, cyclic or acyclic, branched or unbranched carbon chain moiety having the specified number of carbon atoms, or, if not specified, up to 30 carbon atoms. For example, alkyl having 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, as well as moieties that are positional isomers of these moieties. Alkyl having 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, and tetracosyl. In certain embodiments, a straight-chain or branched-chain alkyl has up to 30, and more preferably up to 20, carbon atoms in its backbone (e.g., C1-C6 for a straight chain). 30 , C3-C for branched chains 30 The alkyl group may be substituted or unsubstituted.

[0036] As used herein, the term "heteroalkyl" means an alkyl moiety, as defined above, that contains one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of a carbon atom.

[0037] As used herein, the term "haloalkyl" means an alkyl group, as defined above, substituted with at least one halogen.

[0038] As used herein, the term "hydroxyalkyl" refers to an alkyl group, as defined above, substituted with at least one hydroxyl.

[0039] As used herein, the term "alkylene" refers to an alkyl group having a specified number of carbons, e.g., 2 to 12 carbon atoms, that contains two points of attachment to the remainder of the compound on its longest carbon chain. Non-limiting examples of alkylene groups include methylene-(CH)-, ethylene-(CHCH)-, n-propylene-(CHCHCH)-, isopropylene-(CHCH(CH))-, and the like. Alkylene groups can be cyclic or acyclic, branched or unbranched carbon chain moieties, and can be optionally substituted with one or more substituents.

[0040] "Cycloalkyl" means a monocyclic, bicyclic, bridged, spirocyclic, or polycyclic saturated carbocyclic ring, each of which has from 3 to 12 carbon atoms. Preferred cycloalkyls have from 3 to 10 carbon atoms in their ring structure, and more preferably have from 3 to 6 carbons in the ring structure. Cycloalkyl groups can be substituted or unsubstituted.

[0041] As used herein, the term "halocycloalkyl" refers to a cycloalkyl group, as defined above, that is substituted with at least one halogen.

[0042] "Cycloheteroalkyl" refers to a cycloalkyl moiety, as defined above, that contains one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of a carbon atom. Preferred cycloheteroalkyls have from 4 to 8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4 to 6 carbon and heteroatoms in the ring structure. Cycloheteroalkyl groups can be substituted or unsubstituted.

[0043] Unless the number of carbon atoms is otherwise specified, "lower alkyl," as used herein, refers to an alkyl group, as defined above, having 1 to 10 carbon atoms in its backbone structure, more preferably 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout this application, preferred alkyl groups are lower alkyls. In certain embodiments, a substituent described herein as alkyl is a lower alkyl.

[0044] "Alkenyl" refers to any cyclic or acyclic, branched or unbranched unsaturated carbon chain moiety having a specified number of carbon atoms, or up to 26 carbon atoms if no limit on the number of carbon atoms is specified, and having one or more double bonds within the moiety. Alkenyls having 6 to 26 carbon atoms are exemplified by hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosenyl, docosenyl, tricosenyl, and tetracosenyl, and in various isomeric forms, the unsaturated bond(s) may be at any position within the moiety and may have either the (Z) or (E) configuration about the double bond(s).

[0045] "Alkynyl" refers to a hydrocarbyl moiety within the scope of alkenyl, but having one or more triple bonds within the moiety.

[0046] The term "aryl," as used herein, includes 3- to 12-membered substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon (i.e., carbocyclic aryl) or one or more atoms is a heteroatom (i.e., heteroaryl). Preferably, the aryl group includes a 5- to 12-membered ring, more preferably a 6- to 10-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of which is aromatic; for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Carbocyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, and aniline. Heteroaryl groups include substituted or unsubstituted aromatic 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, and more preferably 5- to 10-membered rings, in which the ring structure contains 1 to 4 heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, etc. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.

[0047] The terms "halo," "halide," or "halogen," as used herein, mean halogens, including, but not limited to, fluoro, chloro, bromo, iodo, and the like, in both radioactive and non-radioactive forms. In preferred embodiments, halo is selected from the group consisting of fluoro, chloro, and bromo.

[0048] The terms "heterocyclyl" or "heterocyclic group" refer to 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, and more preferably 5- to 10-membered rings, which include one to four heteroatoms in the ring structure. The heterocycle can be monocyclic, bicyclic, spirocyclic, or polycyclic. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathine, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like. The heterocyclic ring can be substituted at one or more positions with the substituents described above, such as, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN, etc.

[0049] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more backbone carbons. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound (e.g., one that does not spontaneously undergo transformation by rearrangement, cyclization, elimination, etc.). As used herein, the term "substituted" is intended 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, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituent of organic compounds described herein that satisfy the valence of the heteroatom. The substituents may include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. In a preferred embodiment, the substituent of the substituted alkyl is C 1-6 Alkyl, C 3-6 In a preferred embodiment, the substituent on the substituted alkyl is selected from fluoro, carbonyl, cyano, or hydroxyl. Those skilled in the art will understand that, where appropriate, the substituents themselves may be substituted. Unless specifically stated as "unsubstituted," reference to a chemical moiety herein is understood to include substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0050] As used herein, each expression, e.g., alkyl, m, n, etc., when it occurs more than once in any structure, means that it is independent of its definition elsewhere in the same structure.

[0051] As used herein, "small molecule" means a small organic or inorganic molecule having a molecular weight of less than about 3,000 Daltons. Generally, small molecules useful in the present invention have a molecular weight of less than 3,000 Daltons (Da). A small molecule can be, for example, at least about 100 Da to about 3,000 Da (e.g., about 100 to about 3,000 Da, about 100 to about 2,500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1,500, about 500 to about 1,000, about 300 to about 1,000 Da, or about 100 to about 250 Da).

[0052] In some embodiments, "small molecule" refers to an organic, inorganic, or organometallic compound, typically having a molecular weight of less than about 1000. In some embodiments, small molecules are organic compounds having sizes on the order of 1 nm. In some embodiments, small molecule drugs of the present invention include oligopeptides and other biomolecules having a molecular weight of less than about 1000.

[0053] An "effective amount" is an amount sufficient to achieve a beneficial or desired result. For example, a therapeutic amount is an amount that achieves a desired therapeutic effect. This amount can be the same as or different from a prophylactically effective amount, which is the amount necessary to prevent the onset of a disease or disease symptoms. An effective amount can be administered in one or more administrations, applications, or dosages. The therapeutically effective amount of a composition will vary depending on the composition selected. The composition can be administered once or more times daily to once or more times weekly (including once every other day). One of skill in the art will recognize that certain factors, including but not limited to, the severity of the disease or disorder, previous treatments, the subject's overall health and / or age, and other diseases present, can affect the dosage and timing required to effectively treat a subject. Furthermore, treatment of a subject with a therapeutically effective amount of a composition described herein can include a single treatment or a series of treatments.

[0054] The terms "reduce," "reduce," "reduced," "reduce," "reduce," and "inhibit" are all generally used herein to refer to a statistically significant amount of reduction compared to a reference. However, for the avoidance of doubt, "reduce," "reduce," or "reduce," or "inhibit" typically refers to a reduction of at least 10% compared to a reference level, and can include, for example, a reduction of at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, including any reduction between 10% and 99% compared to the complete absence of a given element or parameter compared to a reference level, or compared to the absence of a given treatment.

[0055] The terms "increased," "increase," or "improve," or "activate" are all used herein to generally mean an increase by a statistically significant amount, and for the avoidance of doubt, the terms "increased," "increase," or "improve," or "activate" mean an increase of at least 10% compared to a reference level, for example, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to and including a 100% increase compared to a reference level, or any increase between 10-100% compared to a reference level, or at least about 2-fold, or at least about 3-fold, or at least about 4-fold, or at least about 5-fold, or at least about 10-fold, or any increase between 2-fold and 10-fold or more compared to a reference level.

[0056] As used herein, the term "modulate" includes upregulation and downregulation, eg, enhancing or inhibiting a response.

[0057] As defined herein, a "radiopharmaceutical agent" refers to a pharmaceutical agent containing at least one radiation-emitting radioisotope. Radiopharmaceutical agents are routinely used in nuclear medicine for the diagnosis and / or treatment of various diseases. Radiolabeled pharmaceutical agents, such as radiolabeled antibodies, contain a radioisotope (RI) that functions as a radiation source. As contemplated herein, the term "radioisotope" includes metallic and non-metallic radioisotopes. The radioisotope is selected based on the medical use of the radiolabeled pharmaceutical agent. When the radioisotope is a metallic radioisotope, a chelating agent is typically used to bind the metallic radioisotope to the remainder of the molecule. When the radioisotope is a non-metallic radioisotope, the non-metallic radioisotope is typically bound to the remainder of the molecule directly or via a linker.

[0058] For purposes of this invention, the chemical elements are identified according to the Periodic Table of the Elements, CAS version, inside pages of Handbook of Chemistry and Physics, 67th Ed., 1986-87.

[0059] Compounds of the Invention One aspect of the present invention is a compound of formula (I): [ka] (In the formula, n is 0 or 1, L1 is absent or selected from -NH-, -N(CH3)-, -O-, and -CH2-; L2 is -alkyl-; L3 is -(5-membered heteroaryl)-; X1 is -C(R1)(R2)(R3), X2 is optionally substituted aryl or heteroaryl; X3 is selected from -H, alkyl, and haloalkyl; R1 is selected from -H, halo, hydroxyl, amido, amino, alkylamino, and aminoalkyl; R2 and R3 are each independently selected from -H and alkyl, or R2 and R3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl; with the proviso that the compound is not selected from: [ka] or a pharmaceutically acceptable salt thereof.

[0060] In certain embodiments, R2 and R3 are each independently selected from -H and alkyl, or R2 and R3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl (not cyclopropyl), or cycloheteroalkyl.

[0061] In certain embodiments, at least one of R1, R2, and R3 is not -H. In other embodiments, at least two of R1, R2, and R3 are not -H. In other embodiments, each of R1, R2, and R3 is not -H.

[0062] In certain embodiments, the compound has the following structure: [ka]

[0063] In certain embodiments, L1 is selected from -NH- and -N(CH3)-.

[0064] In certain embodiments, L2 is selected from -CH2-, -CH2CH2-, and -CH2CH2CH2-.

[0065] In certain embodiments, R1 is -H, -F, -OH, -NH2, -CH2NH2, is selected from -N(H)(CH3), -N(CH3)2, and -C(O)NH2.

[0066] In certain embodiments, R1 is -H. In other embodiments, R1 is -NH2.

[0067] In certain embodiments, R2 and R3 are each -H. In other embodiments, R2 and R3 are each -CH3.

[0068] In certain embodiments, R2 and R3, together with the carbon atom to which they are attached, form an optionally substituted cycloalkyl.

[0069] In certain embodiments, R2 and R3 together with the carbon atom to which they are attached form an unsubstituted cyclopropyl or cyclobutyl.

[0070] In certain embodiments, R2 and R3, together with the carbon atom to which they are attached, form an optionally substituted cycloheteroalkyl.

[0071] In certain embodiments, R2 and R3 together with the carbon atom to which they are attached form an unsubstituted azetidinyl, pyrrolidinyl, piperidinyl, or lactam.

[0072] In certain embodiments, R2 and R3 together with the carbon atom to which they are attached form a substituted azetidinyl, pyrrolidinyl, piperidinyl, or lactam.

[0073] In certain embodiments, the azetidinyl, pyrrolidinyl, piperidinyl, or lactam is N-alkyl or N-acetyl substituted.

[0074] In certain embodiments, X1 is [ka] is selected from.

[0075] In certain embodiments, X1 is [ka] is selected from.

[0076] In certain embodiments, L3 is triazolyl, oxazolyl, or oxadiazolyl.

[0077] In certain embodiments, -L3-X2 is [ka] In other embodiments, -L3-X2 is selected from: [ka] is.

[0078] In certain embodiments, X2 is unsubstituted aryl.

[0079] In certain embodiments, the unsubstituted aryl is unsubstituted phenyl.

[0080] In certain embodiments, X2 is substituted aryl.

[0081] In certain embodiments, the substituted aryl is a substituted phenyl.

[0082] In certain embodiments, X2 is [ka] and R4, R5, R6, R7, and R8 are independently -H, halogen, -CN, -CF3, selected from -CHF2, -OCF3, -OCHF2, alkyl, alkenyl, alkynyl, and cycloalkyl; with the proviso that at least one of R4, R5, R6, R7, and R8 is not -H.

[0083] In certain embodiments, R4, R5, R6, R7, and R8 are independently -H, -Cl, -Br, -F, -CN, -CF3, -OCF3, -CH3, and cyclopropyl; provided that at least one of R4, R5, R6, R7, and R8 is not -H.

[0084] In certain embodiments, X2 is [ka] and R4 is -Cl, -Br, -F, -CN, It is selected from -CF3, -OCF3, -CH3, and cyclopropyl.

[0085] In certain embodiments, X2 is [ka] and R5 is -Cl, -Br, -F, It is selected from -CN, -CF3, -OCF3, -CH3, and cyclopropyl.

[0086] In certain embodiments, X2 is [ka] and R6 is -Cl, -Br, -F, It is selected from -CN, -CF3, -OCF3, -CH3, and cyclopropyl.

[0087] In certain embodiments, X2 is [ka] and R5 and R6 are independently selected from -Cl, -Br, -F, -CN, -CF3, -OCF3, -CH3, and cyclopropyl.

[0088] In certain embodiments, X2 is [ka] and R4 and R6 are independently selected from -Cl, -Br, -F, -CN, -CF3, -OCF3, -CH3, and cyclopropyl.

[0089] In certain embodiments, X2 is [ka] and R5, R6, and R7 are independently selected from -Cl, -Br, -F, -CN, -CF3, -OCF3, -CH3, and cyclopropyl.

[0090] In certain embodiments, X2 is selected from: [ka]

[0091] In certain embodiments, X2 is selected from: [ka]

[0092] In certain embodiments, X3 is -H, -CH3, or -CF3. In other embodiments, X3 is -CF3.

[0093] In certain embodiments, n is 1. In other embodiments, n is 2.

[0094] In certain embodiments, the compound has the following structure: [ka]

[0095] In certain embodiments, the compound has the following structure: [ka]

[0096] In certain embodiments, the compound has the following structure: [ka]

[0097] In certain embodiments, the compound has the following structure: [ka]

[0098] In certain embodiments, the compound has the following structure: [ka]

[0099] In certain embodiments, the compound has the following structure: [ka]

[0100] In some embodiments, the compound is selected from Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] In some embodiments, the compounds are atropisomers. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, replacing a hydrogen with deuterium or tritium, or replacing a carbon with 13 C or 14Compounds produced by replacing the variable R with C-enriched carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or as therapeutic agents according to the present invention. For example, 1 In the case of -C1-C4 alkyl, or -O-(C1-C4) alkyl, the alkyl may be suitably deuterated (e.g., -CD3, -OCD3).

[0101] Any of the compounds of the present invention may also be radiolabeled for the preparation of radiopharmaceuticals.

[0102] Treatment method One aspect of the present invention provides compounds, compositions, and methods useful for treating or preventing diseases or disorders associated with abnormal amino acid concentrations through modulation of SLC6A19 transport.

[0103] Another aspect of the present invention relates to a method of modulating SLC6A19 transport in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0104] Another aspect of the present invention relates to a method of treating or preventing a disease or disorder associated with a genetic defect in phenylalanine hydroxylase in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0105] In certain embodiments, the present invention relates to a method of treating or preventing phenylketonuria in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0106] In certain embodiments, the present invention relates to a method of treating or preventing hyperphenylalaninemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0107] In some embodiments, the compound reduces the subject's systemic phenylalanine concentration.

[0108] In certain embodiments, the present invention relates to a method of treating or preventing tyrosinemia (Type I, Type II, or Type III) in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula (I).

[0109] In certain embodiments, the compound reduces systemic glycine concentrations in a subject.

[0110] In certain embodiments, the present invention relates to a method for treating or preventing isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, a urea cycle disorder, or hyperammonemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0111] In certain embodiments of any one of the disclosed methods, the compound modulates SLC6A19 in the subject.

[0112] In certain embodiments of any one of the disclosed methods, the compound inhibits SLC6A19 in the subject.

[0113] In certain embodiments of any one of the disclosed methods, the compound modulates SLC6A19 trafficking in the subject.

[0114] In certain embodiments of any one of the disclosed methods, the compound inhibits SLC6A19 transport in the subject.

[0115] In certain embodiments, the compound reduces systemic amino acid concentrations in a subject.

[0116] In certain embodiments of any one of the disclosed methods, the subject is a mammal. In certain embodiments of any one of the disclosed methods, the mammal is a human.

[0117] In certain embodiments of any one of the disclosed methods, the compound of formula (I) is defined as follows: [ka] (In the formula, n is 0 or 1, L1 is absent or selected from -NH-, -N(CH3)-, -O-, and -CH2-; L2 is -alkyl-; L3 is -(5-membered heteroaryl)-; X1 is -C(R1)(R2)(R3), X2 is optionally substituted aryl or heteroaryl; X3 is selected from -H, alkyl, and haloalkyl; R1 is selected from -H, halo, hydroxyl, amido, amino, alkylamino, and aminoalkyl; R2 and R3 are each independently selected from -H and alkyl, or R2 and R3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl. Or a pharmaceutically acceptable salt thereof.

[0118] In certain embodiments of any one of the disclosed methods, the compound is selected from the structure of any one of the compounds listed in Table 1.

[0119] In certain embodiments of any one of the disclosed methods, the compound is selected from: [ka]

[0120] In certain embodiments of any one of the disclosed methods, the compound is selected from: [ka] (In the formula, n is 0 or 1, L1 is absent or selected from -NH-, -N(CH3)-, -O-, and -CH2-; L2 is -alkyl-; L3 is -(5-membered heteroaryl)-; X1 is -C(R1)(R2)(R3), X2 is optionally substituted aryl or heteroaryl; X3 is selected from -H, alkyl, and haloalkyl; R1 is selected from -H, halo, hydroxyl, amido, amino, alkylamino, and aminoalkyl; R2 and R3 are each independently selected from -H and alkyl, or R2 and R3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl; with the proviso that the compound is not selected from: [ka] Or a pharmaceutically acceptable salt thereof.

[0121] Pharmaceutical Compositions, Routes of Administration, and Dosage In certain embodiments, the present invention is directed to a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier. In certain embodiments, a pharmaceutical composition comprises multiple compounds of the present invention and a pharmaceutically acceptable carrier.

[0122] In certain embodiments, the pharmaceutical compositions of the present invention further comprise at least one additional pharmaceutically active agent other than a compound of the present invention.

[0123] Pharmaceutical compositions of the present invention can be prepared by combining one or more compounds of the present invention with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents.

[0124] As stated above, "effective amount" refers to any amount sufficient to achieve a desired biological effect. By combining the teachings provided herein and selecting from among various active compounds, and by weighing factors such as potency, relative bioavailability, patient weight, severity of adverse side effects, and method of administration, an effective prophylactic or therapeutic treatment regimen can be designed that is effective in treating a particular subject without causing substantial undesirable toxicity. The effective amount for any particular application may vary depending on factors such as the disease or condition being treated, the particular compound of the present invention being administered, the size of the subject, or the severity of the disease or condition. Those of ordinary skill in the art can empirically determine the effective amount of a particular compound of the present invention and / or other therapeutic agent without necessitating undue experimentation. A maximum dosage, i.e., the highest safe dose according to some medical judgment, can be used. Multiple daily administrations may be contemplated to achieve an appropriate systemic dose of the compound. For example, an appropriate systemic dose can be determined by measuring a patient's peak or sustained plasma concentration of the drug. "Dose" and "administration" are used interchangeably herein.

[0125] In certain embodiments, intravenous administration of the compound may typically be 0.1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 0.1 mg / kg / day to 2 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 0.5 mg / kg / day to 5 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 1 mg / kg / day to 10 mg / kg / day.

[0126] Generally, the daily oral dose of the compound for human subjects is about 0.01 mg / kg / day to 1000 mg / kg / day. Oral doses in the range of 0.5 to 50 mg / kg, administered one or more times per day, are expected to produce therapeutic effects. The dosage can be appropriately adjusted to achieve the desired local or systemic drug concentration, depending on the mode of administration. For example, with intravenous administration, the daily dose is expected to be one to several orders of magnitude lower. If the subject does not respond adequately to such doses, higher doses (or effective high doses via another, more localized delivery route) can be employed, as tolerated by the patient. Multiple daily administrations are contemplated to achieve adequate internal concentrations of the compound.

[0127] For any compound described herein, the therapeutically effective amount can be first determined from animal models.The therapeutically effective amount can also be determined from human data for compounds tested in humans and for compounds known to exhibit similar pharmacological activity, such as other related active agents.Higher doses may be required for parenteral administration.The applied dose can be adjusted according to the relative bioavailability and efficacy of the administered compound.Adjusting the dose to achieve maximum efficacy based on the methods described above and other methods well known in the art is well within the capabilities of those skilled in the art.

[0128] The formulations of the present invention may be administered in pharmaceutically acceptable solutions which may conventionally contain pharmaceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants, and, optionally, other therapeutic ingredients.

[0129] When used in therapy, an effective amount of compound can be administered to a subject by any method that delivers the compound to the desired surface.The administration of pharmaceutical compositions can be carried out by any means known to those skilled in the art.Administration routes include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (bladder), oral, subcutaneous, direct injection (e.g., into tumor or abscess), mucosal (e.g., topical to the eye), inhalation, and topical.

[0130] For intravenous and other parenteral administration routes, the compounds of the present invention can be formulated as lyophilized preparations, as lyophilized preparations of liposome intercalated or liposome-encapsulated active compounds, as lipid complexes in aqueous suspension, or as salt complexes. Lyophilized preparations are generally reconstituted with a suitable aqueous solution, such as sterile water or physiological saline, immediately prior to administration.

[0131] For oral administration, compounds can be easily formulated by combining the active compound(s) with pharmaceutically acceptable carriers well known in the art. Such carriers allow the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by the subject to be treated. Pharmaceutical preparations for oral use can be obtained as solid excipients, optionally by grinding the resulting mixture, and optionally adding suitable excipients, followed by processing the granular mixture to obtain tablets or dragee cores. Suitable excipients are, in particular, sugars, including fillers such as lactose, sucrose, mannitol, or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, e.g., sodium alginate. Optionally, oral preparations may also be formulated with saline or buffers, e.g., EDTA, for neutralizing acidic conditions in the body, or may be administered without any carrier.

[0132] Oral dosage forms of one or more of the components described above are also specifically contemplated. One or more of the components may be chemically modified to facilitate oral delivery of the derivatives. Generally, contemplated chemical modifications involve attaching at least one moiety to the component molecule itself that (a) inhibits hydrolysis and (b) allows uptake from the stomach or intestine into the bloodstream. It may also be desirable to increase the overall stability of one or more of the components and extend their circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, and polyproline. Abuchowski and Davis, "Soluble Polymer-Enzyme Adducts," In: Enzymes as Drugs, Hochenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367-383 (1981); Newmark et al., J. Appl. Biochem. 4:185-9 (1982). Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane. For pharmaceutical applications, as noted above, polyethylene glycol moieties are preferred.

[0133] The location of release of the component (or derivative) may be the stomach, the small intestine (duodenum, jejunum, or ileum), or the large intestine. One skilled in the art has available formulations that will not dissolve in the stomach but will release the substance in the duodenum or elsewhere in the intestine. Preferably, the release will avoid adverse effects in the stomach environment, either by protecting the compound (or derivative) of the invention or by releasing the biologically active substance in the intestine, etc., after passing through the stomach environment.

[0134] To ensure full gastric resistance, a coating that is impermeable to at least pH 5.0 is essential. Examples of more common inactive ingredients used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can also be used as mixed films.

[0135] A coating or mixture of coatings can also be used on tablets not intended for gastric protection. This can include sugar coatings or coatings that make the tablet easier to swallow. Capsules can consist of a hard shell (such as gelatin) for delivery of dry therapeutics (e.g., powder), or a soft gelatin shell can be used for liquid forms. The shell material for cachets can be thick starch or other edible paper. For pills, lozenges, molded tablets, or powder tablets, wet massing techniques can be used.

[0136] The therapeutic agent may also be included in the formulation as fine multiparticulates in the form of granules or pellets about 1 mm in size. The formulation of material for capsule administration may also be as a powder, lightly compressed plugs, or tablets. The therapeutic agent may be prepared by compression.

[0137] Both colorants and flavoring agents may be included. For example, the compounds (or derivatives) of the present invention may be formulated (such as encapsulated in liposomes or microspheres) and then further included in an edible product, such as a refrigerated beverage, containing colorants and flavoring agents.

[0138] The volume of the therapeutic agent can be diluted or increased with an inert material. These diluents can include carbohydrates, especially mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans, and starch. Certain inorganic salts can be used as fillers, including calcium triphosphate, magnesium carbonate, and sodium chloride. Some commercially available diluents include Fast-Flo, Emdex, STA-Rx 1500, Emcompress, and Avicell.

[0139] Disintegrants may be included in the formulation of therapeutic agents to form solid dosage forms. Materials used as disintegrants include, but are not limited to, starch, including the commercially available starch-based disintegrant Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethylcellulose, sponge, and bentonite may also be used. Another form of disintegrant is an insoluble cationic exchange resin. Powdered gums can be used as binders, including powdered gums such as agar, Karaya, or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

[0140] Binders can be used to hold the therapeutic agent and form a hard tablet and include materials derived from natural products such as gum arabic, tragacanth, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can both be used in alcoholic solution to granulate the therapeutic agent.

[0141] Antifriction agents may be included in the formulation of the therapeutic agent to prevent sticking during the formulation process. Lubricants can be used as a layer between the therapeutic agent and the die wall, and include, but are not limited to, stearic acid with magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, and Carbowax 4000 and 6000 can also be used.

[0142] Glidants may be added which may improve the flowability of the drug during formulation and aid in rearrangement during compression. Glidants may include starch, talc, pyrogenic silica, and hydrated silicoaluminate.

[0143] Surfactants may be added as wetting agents to aid in the dissolution of therapeutic agents in aqueous environments. Surfactants may include anionic surfactants such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate, and dioctyl sodium sulfonate. Cationic surfactants may also be used, including benzalkonium chloride and benzethonium chloride. Potential nonionic surfactants that can be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose. These surfactants may be present alone or in a mixture of different ratios in the formulation of the compound or derivative of the present invention.

[0144] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin as well as sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient mixed with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally, stabilizers. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid such as fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All oral formulations must be in a dosage suitable for such administration.

[0145] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0146] For topical administration, the compounds may be formulated as solutions, gels, ointments, creams, suspensions, etc., as is well known in the art. Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, and those designed for transdermal, transmucosal, oral or pulmonary administration.

[0147] For administration by inhalation, the compound for use according to the present invention can be conveniently delivered in the form of aerosol spray presentation from a pressurized pack or nebulizer by using suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.For pressurized aerosol, dosage unit can be determined by providing a valve that delivers a metered amount.For example, gelatin capsules and cartridges for use in inhaler or insufflator can be formulated to contain a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0148] Also contemplated herein is pulmonary delivery of the compounds disclosed herein (or salts thereof). The compounds are delivered to the lungs of a mammal during inhalation, cross the epithelial lining of the lungs, and reach the bloodstream. Other reports on inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprorelin acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl. 5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (α1-proteinase); Oswein et al., 1990, "Aerosolization of Proteins," Proceedings of Symposium on Respiratory Drug Delivery. II, Keystone, Colorado, March (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor-alpha) and U.S. Patent No. 5,284,656 to Platz et al. (granulocyte colony-stimulating factor; incorporated by reference). Methods and compositions for pulmonary delivery of systemically acting drugs are described in U.S. Patent No. 5,451,569, issued September 19, 1995 to Wong et al. (incorporated by reference).

[0149] Contemplated for use in the practice of the present invention are a variety of mechanical devices designed for pulmonary delivery of therapeutic products, including, but not limited to, nebulizers, metered dose inhalers, and dry powder inhalers, all of which are well known to those skilled in the art.

[0150] Some specific examples of commercially available devices suitable for practicing the present invention are the Ultravent nebulizer manufactured by Mallinckrodt, Inc., St. Louis, Mo., the Acorn II nebulizer manufactured by Marquest Medical Products, Englewood, Colo., the Ventolin metered-dose inhaler manufactured by Glaxo Inc., Research Triangle Park, North Carolina, and the Spinhaler powder inhaler manufactured by Fisons Corp., Bedford, Mass.

[0151] All such devices require the use of formulations suitable for dispensing and administering the compounds of the present invention. Typically, each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material in addition to the usual diluents, adjuvants, and / or carriers useful in therapeutics. The use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers is also contemplated. The chemically modified compounds of the present invention can be prepared into various formulations depending on the type of chemical modification or the type of device employed.

[0152] Formulations suitable for use in either jet or ultrasonic nebulizers typically contain a compound (or derivative) of the invention dissolved in water at a concentration of about 0.1 to 25 mg of biologically active compound of the invention per mL of solution. The formulation may also contain a buffer and a simple sugar (e.g., for inhibitor stabilization and to control osmolality). Nebulizer formulations may also contain a surfactant to reduce or prevent surface-induced aggregation of the compound of the invention caused by atomization of the solution to form the aerosol.

[0153] Formulations for use with metered dose inhalers generally comprise a fine powder containing the compound (or derivative) of the present invention suspended in a propellant with the aid of a surfactant. The propellant can be any conventional material employed for this purpose, such as a chlorofluorocarbon, hydrochlorofluorocarbon, hydrofluorocarbon, or hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or a combination thereof. Suitable surfactants include sorbitan trioleate and soybean lecithin. Oleic acid can also be useful as a surfactant.

[0154] Formulations for dispensing from powder inhalation devices comprise a finely divided dry powder containing a compound (or derivative) of the invention, and may contain a bulking agent such as lactose, sorbitol, sucrose, or mannitol in an amount sufficient to facilitate dispersion of the powder from the device, e.g., 50-90% by weight of the formulation. The compound (or derivative) of the invention should advantageously be prepared in particulate form having an average particle size of less than 10 micrometers (μm), most preferably 0.5-5 μm, for most effective delivery to the deep lung.

[0155] Nasal delivery of the pharmaceutical compositions of the present invention is also contemplated. Nasal delivery allows the pharmaceutical compositions of the present invention to enter the bloodstream directly after administration of the therapeutic product to the nose, without the product being deposited in the lungs. Nasal delivery formulations include those based on dextran or cyclodextran.

[0156] For nasal administration, a useful device is a small, hard bottle equipped with a metered-dose sprayer. In one embodiment, the metered dose is delivered by drawing a solution of the pharmaceutical composition of the present invention into a chamber of a fixed volume, which has an opening sized to aerosolize the aerosol formulation by forming a spray when the liquid in the chamber is compressed. The chamber is compressed to administer the pharmaceutical composition of the present invention. In a specific embodiment, the chamber is a piston arrangement. Such devices are commercially available.

[0157] Alternatively, it is a plastic squeeze bottle with an opening or aperture sized to aerosolize the aerosol formulation by forming a spray when squeezed. The opening is usually in the top of the bottle, which is generally tapered to partially fit into the nasal passages for efficient administration of the aerosol formulation. Preferably, the nasal inhaler provides a metered amount of the aerosol formulation to administer a measured amount of drug.

[0158] When systemic delivery is desired, the compound can be formulated for parenteral administration by injection, for example, bolus injection or continuous infusion. Injectable preparations can be provided in unit dosage form, for example, in ampoules or multi-dose containers, with preservatives added. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and can contain formulatory agents such as suspending agents, stabilizers, and / or dispersing agents.

[0159] Pharmaceutical preparations for parenteral administration include aqueous solutions of active compounds in water-soluble form.In addition, suspensions of active compounds can be prepared as suitable oily injection suspensions.Suitable lipophilic solutions or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions can contain substances that increase the viscosity of suspensions, such as sodium carboxymethylcellulose, sorbitol, or dextran.Optionally, suspensions can also contain suitable stabilizers or agents that increase the solubility of compounds, so as to allow the preparation of highly concentrated solutions.

[0160] Alternatively, the active compound may be in powder form for reconstitution with a suitable vehicle, eg, sterile pyrogen-free water, before use.

[0161] The compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.

[0162] In addition to the formulations described above, the compounds can also be formulated as depot preparations. Such long-acting preparations can be formulated using suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as sparingly soluble salts.

[0163] The pharmaceutical compositions may also comprise suitable solid- or gel-phase carriers or excipients, examples of which include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0164] Suitable liquid or solid pharmaceutical preparation forms include, for example, aqueous or saline solutions for inhalation, microencapsulation, cochleation, coating on fine gold particles, encapsulation in liposomes, nebulization, aerosolization, pellets for skin implantation, or dried on sharp objects for rubbing on the skin. Pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or sustained-release preparations of active compounds, which may be prepared using conventional excipients and additives and / or auxiliaries, such as disintegrants, binders, coating agents, swelling agents, lubricants, flavoring agents, sweeteners, or solubilizers, as described above. Pharmaceutical compositions are suitable for use in various drug delivery systems. For a brief review of drug delivery methods, see Langer R, Science 249:1527-33 (1990).

[0165] The compounds of the present invention, and optionally other therapeutic agents, may be administered as is (neat) or in the form of a pharmaceutically acceptable salt or cocrystal. When used in medicine, the salt or cocrystal must be pharmaceutically acceptable; however, pharmaceutically unacceptable salts or cocrystals may conveniently be used to prepare pharmaceutically acceptable salts or cocrystals. Such salts include, but are not limited to, those prepared from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, p-toluenesulfonic acid, tartaric acid, citric acid, methanesulfonic acid, formic acid, malonic acid, succinic acid, naphthalene-2-sulfonic acid, and benzenesulfonic acid. Additionally, such salts may be prepared as alkali metal or alkaline earth metal salts, such as sodium, potassium, or calcium salts of the carboxylic acid group.

[0166] Suitable buffering agents include acetic acid and salts (1-2% w / v), citric acid and salts (1-3% w / v), boric acid and salts (0.5-2.5% w / v), and phosphoric acid and salts (0.8-2% w / v). Suitable preservatives include benzalkonium chloride (0.003-0.03% w / v), chlorobutanol (0.3-0.9% w / v), parabens (0.01-0.25% w / v), and thimerosal (0.004-0.02% w / v).

[0167] The pharmaceutical compositions of the present invention contain an effective amount of a compound described herein and, optionally, a therapeutic agent, contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents, or encapsulating substances suitable for administration to humans or other vertebrates. The term "carrier" refers to a natural or synthetic, organic or inorganic component that is combined with an active ingredient to facilitate application. The components of the pharmaceutical compositions also can be mixed with the compounds of the present invention, and with each other, in a manner such that there is no interaction that would substantially impair the desired pharmaceutical effect.

[0168] The therapeutic agent(s), including but not limited to, the compounds of the present invention, may be provided in particles. As used herein, particle refers to nanoparticles or microparticles (or larger particles in some cases) that may comprise all or part of the compounds of the present invention or other therapeutic agent(s) described herein. The particles may contain the therapeutic agent(s) in a core surrounded by a coating, including but not limited to, an enteric coating. The therapeutic agent(s) may also be dispersed throughout the particle. The therapeutic agent(s) may also be adsorbed onto the particle. The particles may have any order of release rate, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. In addition to the therapeutic agent(s), the particles may contain any material commonly used in the pharmaceutical and medical arts, including but not limited to, disintegrating, non-disintegrating, biodegradable, or non-biodegradable materials, or combinations thereof. The particles may be microcapsules containing the compounds of the present invention in solution or in a semi-solid state. The particles may be of virtually any shape.

[0169] Both non-biodegradable and biodegradable polymeric materials can be used to manufacture particles for delivering therapeutic agent(s). Such polymers can be natural or synthetic. The polymer is selected based on the desired period of release. Bioadhesive polymers of particular interest include the biodegradable hydrogels described in Sawhney HS et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein by reference. These include polyhyaluronic acid, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0170] The therapeutic agent(s) may be contained in a controlled-release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation is controlled. This refers to immediate-release and non-immediate-release formulations, where non-immediate-release formulations include, but are not limited to, sustained-release and delayed-release formulations. The term "sustained-release" (also called "extended-release") is used in its conventional sense to refer to a drug formulation that provides a sustained release of drug over an extended period of time, preferably, but not necessarily, providing a substantially constant blood concentration of drug over an extended period of time. The term "delayed-release" is used in its conventional sense to refer to a drug formulation in which there is a time lag between administration of the formulation and the release of drug from the formulation. "Delayed-release" may or may not involve a sustained release of drug over an extended period of time, and thus may or may not be "sustained-release."

[0171] For the treatment of chronic conditions, the use of long-term sustained-release implants may be particularly suitable. "Long-term" release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and preferably 30 to 60 days. Long-term sustained-release implants are well known to those skilled in the art and include some of the release systems described above.

[0172] It will be understood by those skilled in the relevant art that other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the description of the invention contained herein, in view of the information known to those skilled in the art, and can be made without departing from the scope of the invention or any embodiment thereof. Having now described the invention in detail, the present invention will be more clearly understood by reference to the following examples. The examples are included herein for illustrative purposes only and are not intended to limit the invention. [Example]

[0173] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0174] Example 1: SLC6A19 isoleucine transport assay Cell line generation and maintenance The Flp-In™ T-REx™ 293 cell line was purchased from Thermo Fisher Scientific. This line was used to generate stable cell lines that inducibly express human SLC6A19 containing a C-terminal V5 tag and stably express human TMEM27 (also known as Collectrin) containing a C-terminal myc-DDK tag. Stable cell lines were generated by transfecting plasmids encoding SLC6A19 and TMEM27 using standard protocols, followed by antibiotic selection. Stable cells were maintained in DMEM / F12 supplemented with Glutamax, 10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin, 200 μg / mL hygromycin, 10 μg / mL blasticidin, and 300 μg / mL neomycin (Thermo Fisher).

[0175] Assay: Isoleucine transport assay in 96-well format On day 0, stable cell lines were seeded at a density of 35,000 cells / well in poly-D-lysine-coated, 96-well, cell culture-treated plates. On day 1, SLC6A19 expression was induced by dispensing tetracycline at a final concentration of 1 μg / mL using a Tecan D300e digital dispenser. On day 2, transport assays were performed. The medium was removed from the plate using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio), and the cells were washed with 175 μL of live cell imaging solution (Thermo Fisher Scientific) using the Blue Washer. After washing, cells were treated with either 70 μL of DMSO, positive control, or compound diluted in Krebs buffer (140 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgCl2, 11 mM HEPES, 10 mM glucose, pH 7.4) at room temperature. After 20–60 min, 30 μL of a 3.3 mM solution of 13C6,15N-L-isoleucine (Cambridge Isotope Laboratories) was added. After 20 min of incubation with the isoleucine substrate at room temperature, the cells were washed with 175 μL of live cell imaging solution using a Blue Washer. Cells were then lysed with 150 μL of 15 μM D-Leucine-d10 in ultrapure water (CDN Isotopes). To facilitate lysis, the plate was shaken at 700 rpm for a minimum of 40 min. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the untreated cell lysate. The plate was returned to the shaker for a minimum of 2 min to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000 rpm for 5 min to pellet cell debris and precipitates. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0176] Assay: Isoleucine transport assay in 384-well format On day 0, stable cell lines were seeded at a density of 20,000 cells / well into poly-D-lysine-coated, 384-well cell culture-treated plates containing 1 μg / mL tetracycline using a Viaflo 384-well pipette. Transport assays were performed the following day (day 1). The medium was removed from the plates using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio), and the cells were washed with 80 μL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, cells were treated with either 20 μL of DMSO, positive control, or compound diluted in Krebs buffer (140 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgCl2, 11 mM HEPES, 10 mM glucose, pH 7.4) using a TECAN liquid handler. After incubation at room temperature for 20-60 minutes, 8.6 μL of a 3.3 mM solution of 13C6,15N-L-isoleucine (Cambridge Isotope Laboratories) was added. After incubation with the isoleucine substrate for 20 minutes at room temperature, the cells were washed with 80 μL of live cell imaging solution using a Blue Washer. Cells were then lysed with 1080 μL of 15 μM D-Leucine-d in ultrapure water (CDNIsotopes). To facilitate lysis, the plate was shaken at 700 rpm for a minimum of 2 hours. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the untreated cell lysate. The plate was returned to the shaker for a minimum of 5 minutes to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000 rpm for 10 minutes to pellet cell debris and precipitate. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0177] Analysis of 13C6,15N-L-isoleucine was performed using a RapidFire365-QTOF6545 (Agilent). Quantitative sample analysis utilized automated solid-phase extraction (HILIC H6 cartridge) followed by mass spectrometry injection. Samples were loaded using 95% acetonitrile, 0.1% formic acid and eluted directly from the cartridge using 5% acetonitrile, 0.1% formic acid for ESI-MS (electrospray ionization). Analyte quantification was performed using Agilent Masshunter Quant software from the high-resolution full-scan data.

[0178] Example 2: Synthesis of exemplary compounds Step 1: Synthesis of (S)-1-(tert-butoxycarbonyl)-3-(trifluoromethyl)pyrrolidine-3-carboxylic acid [ka] Step 1: To a mixture of 2-(trifluoromethyl)acrylic acid (15.0 g, 107.1 mmol) and N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methanamine (25.4 g, 107.1 mmol) in DCM (150 mL) was added dropwise TFA (1.22 g, 10.7 mmol) at 0 °C. The resulting mixture was stirred at room temperature for 17 h. The mixture was then diluted with petroleum ether (600 mL), stirred at room temperature for 30 min, and then filtered. The filter cake was then washed with a solution of PE:EtOAc = 3:1 (60 mL) and concentrated in vacuo to give A1 (24.5 g) as a white solid.

[0179] The enantiomers of A1 were separated by chiral SFC (Shimadzu E-UC SFC, CHIRALPAK IC, 5*25 cm, 5 μm) to give (R)-A1 (10.2 g, 34.85% yield) and (S)-A1 (12.1 g, 41.35% yield) as white solids. LC / MS (ESI) m / z: 274 (M+H) + .

[0180] Step 2: To a solution of (S)-A1 (8.0 g, 29.28 mmol) in MeOH (150 mL) was added 20% Pd / C (1.6 g, w / w). The resulting mixture was stirred at room temperature under an H atmosphere for 2 hours. The mixture was then filtered, and the filtrate was concentrated in vacuo to give (S)-3-(trifluoromethyl)pyrrolidine-3-carboxylic acid (5.1 g, 95.12% yield) as a pale yellow oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 184 (M+H) + .

[0181] Step 3: To a mixture of (S)-3-(trifluoromethyl)pyrrolidine-3-carboxylic acid (5.1 g, 27.85 mmol) and TEA (5.64 g, 55.70 mmol) in DCM (70 mL) was added BocO (6.69 g, 30.63 mmol) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated in vacuo, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 12:1) to give 5 (7.5 g, 95.08% yield) as a colorless oil. LC / MS (ESI) m / z: 282 (M−H) - .

[0182] Step 2: Synthesis of aryl amidoximes Following appropriate modification of the following procedures, the aryl amidoximes referred to below can be prepared from appropriately substituted aryl nitriles.

[0183] Synthesis of 4-cyano-N-hydroxybenzimidamide (B1) [ka] To a mixture of terephthalonitrile (12.8 g, 99.90 mmol) and TEA (11.12 g, 109.88 mmol) in EtOH (250 mL) was added hydroxylamine hydrochloride (6.94 g, 99.90 mmol). The resulting mixture was stirred at 70 °C for 2 h. After SM was consumed, the mixture was concentrated in vacuo, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 10:1) to give B1 (11.20 g, 69.57% yield) as a yellow solid. LC / MS (ESI) m / z: 162 (M+H) + .

[0184] Step 3: Synthesis of (S)-3-aryl-5-(3-(trifluoromethyl)pyrrolidin-3-yl)-1,2,4-oxadiazole By appropriate modification of the following procedure, the following 1,2,4-oxadiazoles of type C2 can be prepared from substituted arylamidoxines (eg, B1 and (S)-A2 or racemic A2). [ka]

[0185] Synthesis of 4-cyano-N-hydroxybenzimidamide (C2) Step 1: To a solution of (S)-A2 (7.5 g, 26.48 mmol) in DMF (250 mL) was added CDI (6.44 g, 39.72 mmol). The mixture was stirred at 85 °C for 1 h. Next, B1 (5.50 g, 34.42 mmol) was added to the above mixture. The resulting mixture was stirred at 85 °C for an additional 10 h. After cooling, the mixture was diluted with water (100 mL) and washed twice with EtOAc (60 mL). The combined organic layer was separated, washed with saturated NH4Cl solution (80 mL) and brine (80 mL), dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:0 to 8:1) to give C1 (9.60 g, 88.78% yield) as a colorless oil. LC / MS (ESI) m / z: 409 (M+H) + .

[0186] Step 2: To a 4N HCl / dioxane solution (120 mL) was added C1 (9.60 g, 23.51 mmol) portionwise under N2 atmosphere at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give crude C2 (7.25 g, 89.51% yield) as a pale yellow oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 309 (M+H) + .

[0187] Examples 1-42: By appropriately modifying the following procedure, the compounds in the table below can be prepared from a substituted pyrrolidine (eg, C2) and an appropriate Boc-protected diamine (eg, D1). [ka]

[0188] Step 1: To a mixture of C2 (5.65 g, 28.22 mmol) and DIEA (9.12 g, 70.56 mmol) in DMF (80 mL) was added CDI (4.58 g, 28.22 mmol) at 0 °C. The mixture was stirred at room temperature for 30 min. Then, D1 (7.25 g, 23.51 mmol) was added to the above mixture. The resulting mixture was stirred at 50 °C for 3 h. After cooling, the mixture was quenched with aqueous HCl (120 mL, 2 N) and extracted twice with EtOAc (100 mL). The combined organic layer was separated, washed with brine (120 mL), dried over anhydrous Na2SO4, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 18:1) to give D2 (11.6 g, 92.27% yield) as an off-white solid. LC / MS (ESI) m / z: 435 (M-100+H) + .

[0189] Step 2: To a 4N HCl / dioxane solution (120 mL) at 0 °C under a N atmosphere was added D2 (11.6 g, 21.7 mmol). The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give the crude product (10.6 g) as a pale yellow oil. The crude product was then diluted with MTBE (100 mL) and stirred at room temperature for 5 h, after which the resulting slurry was filtered. The filter cake was washed with MTBE (100 mL) and concentrated under vacuum to give D3 (7.10 g, 75.31% yield) as a white solid. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Table 2-11] [Table 2-12] [Table 2-13] [Table 2-14] [Table 2-15] [Table 2-16] [Table 2-17] [Table 2-18] [Table 2-19] [Table 2-20]

[0190] Examples 43-66: The compounds in the following table can be prepared from substituted pyrrolidines (e.g., E2) prepared according to Procedure 3, or from appropriate diamines (e.g., E1) following appropriate modification of the following procedure. [ka]

[0191] Step 1: To a mixture of E2 (156 mg, 0.440 mmol) and DIEA (74 mg, 0.572 mmol) in DMF (1 mL) at 0 °C, CDI (82 mg, 0.572 mmol) was added portionwise. The reaction mixture was stirred at room temperature for 40 min. Next, E1 (88 mg, 0.88 mmol) was added to the above mixture, and the resulting mixture was stirred at 50 °C for 3 h. The mixture was then concentrated to dryness. The residue was purified by preparative reverse-phase HPLC to give E3 as a colorless oil. LC / MS (ESI) m / z: 444 (M+H) + . [ka] [Table 3-1]

Table 3-2

Table 3-3

Table 3-4

Table 3-5

Table 3-6

Table 3-7

Table 3-8

Table 3-9

Table 3-10

Table 3-11

[0192] Examples 67 - 73: By appropriately modifying the following procedures, the compounds in the table below can be prepared from substituted pyrrolidine (e.g., C2) and appropriate diamine (e.g., F1).

Chemical formula

[0193] Step 1: To a mixture of C2 (40 mg, 0.19 mmol) and DIEA (34 mg, 0.259 mmol) in DMF (0.5 mL) was added CDI (33 mg, 0.234 mmol). The reaction mixture was stirred at room temperature for 15 min. Next, F1 (40 mg, 0.194 mmol) was added to the above mixture, and the resulting mixture was stirred at 50 °C for 3 h. The mixture was then concentrated to dryness. The residue was dissolved in 4 N HCl / dioxane solution (1 m) and stirred for 30 min, then concentrated to dryness again to give F2, which was used without further purification. LC / MS (ESI) m / z: 439 (M+H) + .

[0194] Step 2: To a solution of F2 HCl (43 mg, 98 μmol) in methanol (1.5 mL) was added DIPEA (38 mg, 292 μmol) and 37% formaldehyde solution (22 μL, 292 μmol). Sodium triacetoxyborohydride (42 mg, 195 μmol) was added and the solution was stirred at room temperature. Further portions of sodium triacetoxyborohydride were added until LCMS indicated complete consumption of the starting material. The resulting solution was directly purified by preparative reverse-phase HPLC to give F3 as a colorless oil. LC / MS (ESI) m / z: 453 (M+H) + . [Table 4-1] [Table 4-2] [Table 4-3]

[0195] Examples 74-76: In a manner similar to that used to prepare the pyrrolidines above, the compounds in the following table can be prepared from 1-(tert-butoxycarbonyl)-4-(trifluoromethyl)piperidine-4-carboxylic acid, the appropriate diamine, and the appropriate aryl nitrile. [Table 5-1] [Table 5-2]

[0196] Step 3: Synthesis of (S)-3-aryl-5-(3-(trifluoromethyl)pyrrolidin-3-yl)-1,2,4-oxadiazole 1,2,4-Oxadiazoles of type G5 referred to below can be prepared from nitriles G1 following appropriate modification of the following procedure, and from appropriately substituted benzoates (eg G3) following the following procedure. [ka]

[0197] Step 1: To a solution of G1 (1.07 g, 4.21 mmol) in ethanol (10 mL) was added hydroxylamine hydrochloride (585 mg, 8.42 mmol) and triethylamine (1.17 mL, 8.42 mmol). The solution was heated at 60 °C for 2 h and then cooled to room temperature. The solution was diluted with water and ethyl acetate, and the phases were separated. The aqueous phase was extracted twice more with ethyl acetate, and the combined organic phases were then washed with water, saturated aqueous NaCl, dried over MgSO4, and concentrated to give G2 (1.3 g, 4.53 mmol) as a white solid, which was used without further purification. LC / MS (ESI) m / z: 288.0 (M+H) + .

[0198] Step 2: To a mixture of G2 (324 mg, 1.13 mmol) and G3 (176 mg, 1.13 mol) in dioxane (3 mL) at room temperature, DCC (256 mg, 1.24 mmol) was added. The solution was then heated at 90 °C overnight and then cooled to room temperature. The precipitate was removed by filtration, and the filtrate was concentrated to an oil, which was purified by column chromatography (0-100% ethyl acetate / heptane) to give the desired oxadiazole G4 (125.4 mg) as a colorless residue. LC / MS (ESI) m / z: 399.1 (M+H) + .

[0199] Step 3: To a solution of G4 (125.4 mg, 0.294 mmol) in DCE (1 mL) at room temperature was added 1-chloroethyl chloroformate (67 uL, 0.614 mmol). The solution was stirred at 50° C. for 75 minutes, then methanol (1 mL) was added and the solution was heated at 60° C. for 1 hour. The solution was then concentrated to give G5 HCl, which was used without further purification. LC / MS (ESI) m / z: 309.1 (M+H) + .

[0200] Examples 77-79: The compounds in the table below can be prepared from 1,2,4-oxadiazoles of type G5 and the appropriate diamines following appropriate modifications of the methods reported above. [Table 6-1] [Table 6-2]

[0201] Example 80: Example 80 was prepared according to the following method. [ka]

[0202] Step 1: To a mixture of A1 (210 mg, 0.77 mmol) in DMF (5 mL), DIPEA (199 mg, 1.54 mmol) and 2-bromo-1-(4-bromophenyl)ethan-1-one (213 mg, 0.77 mmol) were added. The resulting mixture was stirred at room temperature for 3 h. The mixture was then diluted with water (20 mL) and extracted twice with EtOAc (15 mL). The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by silica gel column chromatography (eluted with PE:EtOAc = 100:0 to 40:1) to give H1 (350 mg, 97.01% yield) as a colorless oil. LC / MS (ESI) m / z: 470 / 472 (M+H)+ .

[0203] Step 2: To a solution of H1 (350 mg, 0.75 mmol) in toluene (15 mL), acetamide (1.28 g, 21.75 mmol) and boron trifluoride diethyl etherate (1.5 mL, 0.73 mmol) were added. The resulting mixture was stirred in a sealed tube at 150 °C for 6 h. The reaction was continued until TLC showed complete consumption of the starting material (PE: EtOAc = 10:1). The mixture was then quenched with saturated NaHCO3 solution (40 mL) at 0 °C and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE: EtOAc = 100:0 to 100:3) to give H2 (135 mg, 40.20% yield) as a colorless oil. LC / MS (ESI) m / z: 451 / 453 (M+H). + .

[0204] Step 3: To a mixture of H2 (135 mg, 0.30 mmol) and Zn(CN)2 (53 mg, 0.45 mmol) in DMF (7 mL) was added Pd(PPh3)4 (35 mg, 0.03 mmol), and the resulting mixture was stirred at 120 °C under a N2 atmosphere for 16 h. The mixture was then diluted with H2O (20 mL) and extracted twice with EtOAc (15 mL). The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by silica gel column chromatography (eluted with PE:EtOAc = 100:1 to 4:1) to give H3 (83 mg, 69.82% yield) as a colorless oil. LC / MS (ESI) m / z: 398 (M+H) + .

[0205] Step 4: To a solution of H3 (83 mg, 0.21 mmol) in DCM (5 mL) was added chloroethyl chloroformate (90 mg, 0.63 mmol), and the resulting mixture was stirred at 50 °C for 20 h under a N2 atmosphere. Next, MeOH (3 mL) was added to the above mixture, and the mixture was stirred at 75 °C for an additional 2 h. The mixture was then concentrated to give crude H4 HCl (64 mg, 99.71% yield) as a colorless oil. LC / MS (ESI) m / z: 308 (M+H) + .

[0206] Step 5: To a solution of D1 HCl (55 mg, 0.27 mmol) in DMF (4 mL), DIEA (76 mg, 0.59 mmol) and CDI (44 mg, 0.27 mmol) were added, and the mixture was stirred at 0 °C for 30 min. Next, H4 HCl (64 mg, 0.21 mmol) was added to the above mixture, and the resulting mixture was stirred at 50 °C for an additional 2 h. The mixture was then diluted with HO (20 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 100:3) to give Boc-F5 (70 mg, 63.02% yield) as a colorless oil. LC / MS (ESI) m / z: 534 (M+H) + To a round-bottom flask was added Boc-H5 (70 mg, 0.13 mmol) and HCl / dioxane (4 mol / L, 4 mL) at 0 °C. The resulting mixture was stirred at room temperature for 1 h. LCMS showed complete consumption of the starting material. The mixture was then concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC to give H5 (39.0 mg, 68.55% yield) as a white solid. [Table 7]

[0207] Example 81: Example 81 was prepared according to the following method. [ka]

[0208] Step 1: To a mixture of (rac)-A2 (5.18 g, 18.29 mmol) and DIEA (7.09 g, 54.86 mmol) in MeCN (100 mL) was added CDI (4.45 g, 27.43 mmol) at 0 °C. The mixture was stirred at 90 °C for 1 h. Then, N,O-dimethylhydroxylamine hydrochloride (2.32 g, 23.77 mmol) was added to the above mixture. The resulting mixture was stirred at 90 °C for another 3 h. After cooling, the mixture was quenched with aqueous HCl (100 mL, 2 N) and extracted twice with dichloromethane (100 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous NaSO, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE: EtOAc = 100:0 to 4:1) to give I1 (5.09 g, 85.29% yield) as a white solid. LC / MS (ESI) m / z: 271 (M-56+H) + .

[0209] Step 2: To a solution of I1 (2.0 g, 6.13 mmol) in dry DCM (50 mL) was added a solution of DIBAL-H in THF (18.4 mL, 1 M) dropwise at -78 °C under a N2 atmosphere. The resulting mixture was stirred at -78 °C for 1 h. The mixture was then quenched portionwise with saturated potassium sodium tartrate tetrahydrate solution (50 mL) at 0 °C and stirred at room temperature for 1 h. The mixture was then extracted twice with DCM (60 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated to dryness under reduced pressure. The crude product was purified by silica gel column chromatography (eluted with PE:EtOAc = 100:0 to 20:1) to give I2 (1.50 g, 91.58% yield) as a white solid. LC / MS (ESI) m / z: 212 (M-56+H). + .

[0210] Step 3: To a solution of I2 (1.50 g, 5.61 mmol) in MeOH (50 mL), K2CO3 (2.33 g, 16.84 mmol) and (1-diazo-2-oxopropyl)phosphonic acid dimethyl ester (1.62 g, 8.42 mmol) were added. The resulting mixture was stirred at room temperature for 2 h. The mixture was then diluted with water (80 mL) and extracted twice with MTBE (80 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by silica gel column chromatography (eluted with PE:EtOAc = 100:0 to 40:1) to give I3 (1.25 g, 84.60% yield) as a white solid.

[0211] Step 4: To a mixture of 4-azidobenzonitrile (70 mg, 0.48 mmol) and I3 (140 mg, 0.53 mmol) in DMSO (6 mL) at room temperature, CuSO4 (8 mg, 0.05 mmol) and sodium ascorbate (77 mg, 0.39 mmol) were added. The resulting mixture was stirred at 60 °C under a N2 atmosphere for 3 h. The mixture was then diluted with water (30 mL) and extracted twice with DCM (15 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (eluted with PE:EtOAc = 100:0 to 4:1) to give Boc-I4 (94 mg, 47.51% yield) as a yellow oil. LC / MS (ESI) m / z: 408 (M+H) + To a vessel containing .Boc-I4 (94 mg, 0.23 mmol) was added 4N HCl / dioxane solution (5 mL) at 0 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give crude I4 HCl (75 mg, 95.07% yield) as a pale yellow oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 308 (M+H) + .

[0212] Step 5: To a mixture of D1 HCl (59 mg, 0.29 mmol) and DIPEA (95 mg, 0.73 mmol) in DMF (8 mL) was added CDI (47 mg, 0.29 mmol) at 0 °C. The mixture was stirred at room temperature for 1 h. Then, I4 HCl (75 mg, 0.23 mmol) was added to the above mixture, and the resulting mixture was stirred at 40 °C for an additional 3 h. After cooling, the mixture was quenched with saturated aqueous NaHCO3 solution (20 mL) and extracted twice with EtOAc (15 mL). The combined organic layers were washed with saturated NH4Cl solution (30 mL) and brine (30 mL), dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 15:1) to afford Boc-I5 (55 mg, 42.23% yield) as an off-white solid. LC / MS (ESI) m / z: 434 (M-100+H) + .Boc-I5 (55 mg, 0.10 mmol) was added portionwise to a 4 N HCl / dioxane solution (5 mL) at 0 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC to give I5 (18 mg, 40.29% yield) as a colorless oil. [Table 8]

[0213] Example 82: Example 82 was prepared according to the following method. [ka]

[0214] To a solution of J1 (15 mg, 53 μmol) and 5-((tert-butoxycarbonyl)amino)pentanoic acid (15 mg, 69 μmol) in DMF (0.5 mL) was added DIPEA (28.5 μL, 106 μmol) and HATU (26.3 mg, 69 μmol). The solution was stirred at room temperature and then concentrated to give a residue which was dissolved in 4N HCl / dioxane (1 mL). The solution was kept at room temperature and then concentrated to a residue. The product was dissolved in methanol and purified by preparative reverse-phase HPLC to give J2. [Table 9]

[0215] Example 83: Example 83 was prepared according to the following method. [ka]

[0216] Step 1: To a solution of (rac)-A2 (108 mg, 381 umol) in THF (2 mL) was added 4-chlorobenzoic acid hydrazide (65.1 mg, 381 umol), triethylamine (159 uL, 1.14 mmol), and T3P (587 mg, 953 umol). The solution was heated at 75 °C for 3 hours and then at room temperature overnight. The solution was poured into water, and the aqueous phase was extracted twice with ethyl acetate. The combined organic phase was washed twice with saturated aqueous sodium bicarbonate, then once with brine, dried over MgSO4, filtered, and concentrated to a residue that was purified by column chromatography to give H1 as a white solid (117 mg, 270 umol). LC / MS (ESI) m / z: 335.9 (M-100+H). + .

[0217] Step 2: To a solution of H1 (118 mg, 268 umol) in MeCN (1 mL) was added DIPEA (94 uL, 547 umol) and pTsCl (202 mg, 536 umol). The solution was stirred at room temperature for 3 hours and then diluted with water and ethyl acetate. The phases were separated and the organic phase was washed with saturated aqueous NaHCO3, dried over MgSO4, filtered, and concentrated. The product was purified by column chromatography to give H2 (85 mg, 203 umol) as a colorless oil. LC / MS (ESI) m / z: 362.2 (M-57+H) + .

[0218] Step 3: HCl / dioxane (1 mL) was added to a vial containing H2 (85 mg, 203 umol). The solution was kept at room temperature for 30 minutes, then H3 was obtained as a colorless residue, which was used without further purification.

[0219] Step 4a: To a suspension of CDI (2.11 g, 14.68 mmol) in DCM (10 mL) at 0 °C, E1 (974 mg, 9.72 mmol) was added. The solution was stirred at room temperature for 15 min, concentrated to a residue, and purified by column chromatography (0 to 100% DCM / methanol). The product H4 (1.74 g) was recovered as a viscous oil.

[0220] Step 4b: To a solution of H3 (40 mg, 113 μmol) and DIPEA (40 μL, 225 μmol) in DMF (0.5 mL) was added H4 (44 mg, 225 μmol). The solution was heated at 50° C. for 2 h and then cooled to room temperature. The reaction mixture was directly purified by reverse-phase preparative HPLC to give H5 as a colorless oil. [Table 10]

[0221] Incorporation by Reference All US patents and US patent application publications cited herein are hereby incorporated by reference.

[0222] equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

1. Compounds of formula (I): 【Chemical 1】 (In the formula, n is 0 or 1; L １ is absent or is -NH-, -N(CH ３ )-, -O-, and -CH ２ - is selected from, L ２ is -alkyl-, L ３ is -(5-membered heteroaryl)-; X １ is -C(R １ ) (R ２ ) (R ３ ) and X ２ is an optionally substituted aryl or heteroaryl; X ３ is selected from —H, alkyl, and haloalkyl; R １ is selected from —H, halo, hydroxyl, amido, amino, alkylamino, and aminoalkyl; R ２ and R ３ are each independently selected from —H and alkyl, or R ２ and R ３ together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl; with the proviso that the compound is not selected from: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1 having the following structure: 【Chemistry 3】

3. L １ is -NH- and -N(CH ３ 3. The compound according to claim 1, wherein the compound is selected from the group consisting of:

4. L ２ But -CH ２ -, -CH ２ CH ２ - and -CH ２ CH ２ CH ２ The compound according to any one of claims 1 to 3, selected from:

5. R １ -H, -F, -OH, -NH ２ , -CH ２ NH ２ , -N(H)(CH ３ ) ２ , -N(CH ３ ) ２ and —C(O)NH ２ The compound according to any one of claims 1 to 4, selected from:

6. R １ The compound of claim 5, wherein is —H.

7. R １ But -NH ２ 6. The compound of claim 5, wherein:

8. R ２ and R ３ The compound according to any one of claims 1 to 7, wherein each is -H.

9. R ２ and R ３ are -CH ３ The compound according to any one of claims 1 to 7,

10. R ２ and R ３ The compound of any one of claims 1 to 7, wherein together with the carbon atom to which they are attached form an optionally substituted cycloalkyl.

11. R ２ and R ３ together with the carbon atom to which they are attached form an unsubstituted cyclopropyl or cyclobutyl.

12. R ２ and R ３ The compound of any one of claims 1 to 7, wherein together with the carbon atom to which they are attached form an optionally substituted cycloheteroalkyl.

13. R ２ and R ３ together with the carbon atom to which they are attached form an unsubstituted azetidinyl, pyrrolidinyl, piperidinyl, or lactam.

14. R ２ and R ３ together with the carbon atom to which they are attached form a substituted azetidinyl, pyrrolidinyl, piperidinyl, or lactam.

15. 15. The compound of claim 14, wherein the azetidinyl, pyrrolidinyl, piperidinyl, or lactam is N-alkyl or N-acetyl substituted.

16. X １ but, 【Chemistry 4】 The compound according to any one of claims 1 to 15, selected from:

17. L ３ The compound of any one of claims 1 to 16, wherein is triazolyl, oxazolyl, or oxadiazolyl.

18. -L ３ -X ２ but, 【Chemistry 5】 The compound according to any one of claims 1 to 17, selected from:

19. -L ３ -X ２ but, 【Chemistry 6】 The compound according to any one of claims 1 to 17,

20. X ２ The compound of any one of claims 1 to 19, wherein is unsubstituted aryl.

21. 21. The compound of claim 20, wherein the unsubstituted aryl is unsubstituted phenyl.

22. X ２ The compound of any one of claims 1 to 19, wherein is substituted aryl.

23. 23. The compound of claim 22, wherein the substituted aryl is a substituted phenyl.

24. X ２ but, 【Chemistry 7】 and R ４ , R ５ , R ６ , R ７ , and R ８ are independently —H, halogen, —CN, or —CF ３ , -CHF ２ , -OCF ３ , -OCHF ２ , alkyl, alkenyl, alkynyl, and cycloalkyl; ４ , R ５ , R ６ , R ７ , and R ８ The compound of claim 23, wherein at least one of: is not —H.

25. R ４ , R ５ , R ６ , R ７ , and R ８ are independently —H, —Cl, —Br, —F, —CN, or —CF ３ , -OCF ３ , -CH ３ and cyclopropyl; ４ , R ５ , R ６ , R ７ , and R ８ 25. The compound of claim 24, wherein at least one of: is not —H.

26. X ２ but, 【Chemistry 8】 and R ４ is -Cl, -Br, -F, -CN, -CF ３ , -OCF ３ , -CH ３ and cyclopropyl.

27. X ２ but, 【Chemistry 9】 and R ５ is -Cl, -Br, -F, -CN, -CF ３ , -OCF ３ , -CH ３ and cyclopropyl.

28. X ２ but, 【Chemistry 10】 and R ６ is -Cl, -Br, -F, -CN, -CF ３ , -OCF ３ , -CH ３ and cyclopropyl.

29. X ２ but, 【Chemistry 11】 and R ５ and R ６ are independently —Cl, —Br, —F, —CN, or —CF ３ , -OCF ３ , -CH ３ and cyclopropyl.

30. X ２ but, 【Chemistry 12】 and R ４ and R ６ are independently —Cl, —Br, —F, —CN, or —CF ３ , -OCF ３ , -CH ３ and cyclopropyl.

31. X ２ but, 【Chemistry 13】 and R ５ , R ６ , and R ７ is -Cl, -Br, -F, -CN, -CF ３ , -OCF ３ , -CH ３ 24. The compound of claim 23, wherein the aryl group is independently selected from:

32. X ２ The compound according to any one of claims 1 to 25, wherein is selected from the following: 【Chemistry 14】

33. X ３ But -H, -CH ３ , or -CF ３ The compound according to any one of claims 1 to 32,

34. X ３ But, -CF ３ The compound according to any one of claims 1 to 33,

35. The compound of any one of claims 1 to 34, wherein n is 1.

36. The compound of any one of claims 1 to 34, wherein n is 2.

37. 36. The compound of claim 35 having the following structure: 【Chemistry 15】

38. 36. The compound of claim 35 having the following structure: 【Chemistry 16】

39. 36. The compound of claim 35 having the following structure: 【Chemistry 17】

40. 37. The compound of claim 36 having the following structure: 【Chemistry 18】

41. 37. The compound of claim 36 having the following structure: 【Chemistry 19】

42. 37. The compound of claim 36 having the following structure: 【Chemistry 20】

43. A compound having the structure of any one of the compounds listed in Table 1, or a pharmaceutically acceptable salt thereof.

44. A pharmaceutical composition comprising a compound according to any one of claims 1 to 43 and a pharmaceutically acceptable excipient.

45. 44. A method for treating or preventing a disease or disorder associated with a genetic defect in phenylalanine hydroxylase, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 43.

46. 44. A method for treating or preventing phenylketonuria, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 43.

47. 44. A method for treating or preventing hyperphenylalaninemia, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 43.

48. 48. The method of any one of claims 45 to 47, wherein the compound reduces systemic phenylalanine concentrations in the subject.

49. 44. A method of treating or preventing tyrosinemia (Type I, Type II, or Type III), comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1 to 43.

50. 50. The method of claim 49, wherein the compound reduces systemic tyrosine levels in the subject.

51. 44. A method for treating or preventing nonketotic hyperglycinemia, comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1 to 43.

52. 52. The method of claim 51, wherein the compound reduces systemic glycine concentrations in the subject.

53. 44. A method for treating or preventing isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, a urea cycle disorder, or hyperammonemia, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 43.

54. 44. A method for treating or preventing diabetes, chronic kidney disease, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, metabolic syndrome, obesity-related disorders, or neurodevelopmental disorders and autism spectrum disorders, comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1 to 43.

55. 55. The method of any one of claims 45 to 54, wherein the compound inhibits SLC6A19 in the subject.

Images