Small molecule inhibitors of mammalian SLC6A19 function

Abstract

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

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 292,818, filed December 22, 2021. [Background technology]

[0002] Phenylketonuria (PKU) is an inborn error of metabolism caused by mutations in phenylalanine hydroxylase (PAH), an enzyme involved in metabolizing phenylalanine. PKU is an autosomal recessive metabolic disorder in which phenylalanine is not properly metabolized, resulting in abnormally high levels of plasma phenylalanine. People with PKU have abnormally high blood levels of phenylalanine, which, if untreated, can lead to irreversible neurological damage resulting in a wide variety of complications, including intellectual disability, seizures, neurodevelopmental disorders, and behavioral disorders. PKU is difficult to treat because blood levels of phenylalanine are directly related to diet. Patients must adhere to strict lifelong dietary habits that affect all aspects of the patient's life. The current standard of care is enzyme cofactor and enzyme replacement therapy, but these therapies are not effective for 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 at chromosome 12q23.2 are known to cause most forms of PKU. These LOF mutations resulting in PKU can be diagnosed as classical PKU (the most severe form), and "mild PKU" or "hyperphe", which are less severe forms. 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 result in elevated phenylalanine levels. In addition to diet, blood amino acid levels, including phenylalanine, are regulated by SLC6A19, which is located in the proximal tubules of the kidney and is involved in the reabsorption of amino acids back into the blood. Summary of the Invention [Means for solving the problem]

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

[0005] Thus, a compound having the structure of formula (I): [ka] During the ceremony, L1 is absent or selected from -alkyl-, -cycloalkyl-, and -heteroaryl-CH2-; X1 and X2 are independently selected from -H, alkyl, cycloalkyl, and alkyl-cycloalkyl, with the proviso that X1 and X2 are not both -H; Y1 is optionally substituted aryl; Y2 is selected from optionally substituted pyridonyl, optionally substituted pyrimidinoyl, optionally substituted pyrazinonyl, optionally substituted triazinonyl, and optionally substituted quinazolinonyl; Y3, Y4, Y5, and Y6 are independently selected from -H and a halide; Provided herein are compounds, or pharma- ceutically acceptable salts thereof.

[0006] Another aspect of the 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).

[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, urea cycle disorders, or hyperammonemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0008] Another aspect of the invention relates to a method of modulating SLC6A19 trafficking in a subject in need thereof, the method 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 those skilled 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 take precedence. In addition, 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 be apparent from the detailed description and claims. [Brief description of the drawings]

[0011] [Figure 1] 1 is a table summarizing isoleucine transport data for exemplary compounds of the invention. A=IC50<500nM, B=IC50 500nM-1,500nM, C=IC50>1,500nM-5,000nM, and D=IC50>5,000nM-10,000nM. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

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

[0013] In order that the present invention may be more readily understood, 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] In this specification and in the claims, the phrase "and / or" as used therein should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with "and / or" should be interpreted in the same manner, i.e., "one or more" of the elements so conjoined. Other elements other than the elements specifically identified by the "and / or" clause may optionally be present, whether or not related to those elements specifically identified. Thus, as a non-limiting example, a reference to "A and / or B", when used in conjunction with open-ended language such as "comprising", may refer in one embodiment to only A (optionally including elements other than B), in another embodiment to only B (optionally including elements other than A), in yet another embodiment to both A and B (optionally including other elements), etc.

[0016] As used herein and in the claims, "or" should 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., the inclusion of at least one, but also including more than one of a number or list of elements, and optionally additional non-listed items. When used only with terms clearly indicated to the contrary, such as "only one of" or "exactly one of," or in the claims, "consisting of" refers to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein 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 and in the claims, the phrase "at least one," with reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of 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 elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, may optionally be present, whether or not related 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 A, optionally including multiples, with no B present (and optionally including elements other than B), in another embodiment to at least one B, optionally including multiples, with no A present (and optionally including elements other than A), in yet another embodiment to at least one A, optionally including multiples, and at least one B, optionally including multiples (and optionally including other elements), etc.

[0018] It is also to be understood that, unless expressly indicated to the contrary, in any method claimed herein that includes multiple steps or acts, the order of the method steps or acts is not necessarily limited to the order in which the method steps or acts are recited.

[0019] In the claims and in the above specification, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "consisting of," and the like, are to be understood as open ended, i.e., meaning inclusive without limitation. Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as set forth 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 specific geometric or stereoisomeric forms. 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 thereof. 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" means an isomer that differs in the orientation of a substituent atom in relation to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. The atoms (other than H) on each side of a carbon-carbon double bond may be in the E (substituents are on opposite sides of the carbon-carbon double bond) or Z (substituents are oriented on the same side) configuration. "R", "S", "S*", "R*", "E", "Z", "cis", and "trans" refer to configurations relative to the core molecule. Certain disclosed compounds may exist in "atropisomeric" forms or as "atropisomers". Atropisomers are stereoisomers that result from the steric strain barrier to rotation being high enough to permit isolation of conformers, and rotation around a single bond being hindered. The compounds of the present invention may be prepared as individual isomers by isomer-specific synthesis or by separation from a mixture of isomers. Traditional separation techniques include forming salts of the free bases of each isomer of the isomeric pair using optically active acids (followed by fractional crystallization and regeneration of the free bases), forming salts of the acid forms of each isomer of the isomeric pair using optically active amines (followed by fractional crystallization and regeneration of the free acids), forming esters or amides of each of the isomers of the isomeric pair using optically pure acids, amines, or alcohols (followed by chromatographic separation and removal of the chiral auxiliary), or separating isomeric mixtures of either the starting materials or the final products using a variety of well-known chromatographic methods.

[0022] For example, if a specific enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, the resulting diastereomeric mixture separated, and the auxiliary group cleaved to provide the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, a diastereomeric salt is formed with a suitable optically active acid or base, followed by separation of the formed diastereomers by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomer.

[0023] The term "tautomer" as used herein refers to a structural isomer that exists in equilibrium resulting from the migration of a hydrogen atom. For example, two tautomers of 2-pyrimidinone are listed below. Only one tautomer may be provided in the structural representation of a given compound. However, the present invention contemplates all such tautomers of a given compound. [ka]

[0024] Percent purity by mole fraction is the ratio of moles of an enantiomer (or diastereomer), or the ratio over moles of an enantiomer (or diastereomer) plus the 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 compared to the other stereoisomer. When only one 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 only one 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.

[0025] 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 either an enantiomer of the compound without the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer compared to its corresponding optical isomer.When a disclosed compound is named or depicted by structure without indicating stereochemistry and has two or more chiral centers, the name or structure should be understood to encompass a diastereomer without the other diastereomer, multiple diastereomers without other diastereomeric pairs, a mixture of diastereomers, a mixture of diastereomeric pairs, a mixture of diastereomers enriched in one diastereomer compared to the other diastereomer(s), or a mixture of diastereomers enriched in one or more diastereomers compared to the other diastereomers.The present invention encompasses all of these forms.

[0026] Structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, by the replacement of hydrogen with deuterium or tritium, or of carbon with 13 C or 14 Compounds produced by replacing C with C-enriched carbon are within the scope of the present invention.

[0027] The term "prodrug" as used herein includes compounds that are converted to therapeutically active agents under physiological conditions. A common method for making a prodrug is to include a selected moiety that is hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of the host animal.

[0028] As used herein, the phrase "pharmacologically acceptable excipient" or "pharmacologically acceptable carrier" means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, involved in carrying or transporting the subject chemical 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 harmful to the patient, and substantially non-pyrogenic. Some examples of materials which can function as pharma- ceutically 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 the like. and 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 used in pharmaceutical formulations. In certain embodiments, the pharmaceutical compositions of the present invention are non-pyrogenic, i.e., do not induce a significant increase in body temperature when administered to a patient.

[0029] 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 compound(s) purified to a free base form with a suitable organic or inorganic acid and isolating the salt formed thereby. Representative salts include hydrobromide, hydrochloride, sulfate, hydrogen sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulfonate. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J.Pharm.Sci.66:1-19.)

[0030] In other cases, compounds useful in the methods of the present invention may contain one or more acidic functional groups and therefore can form pharma-ceutically acceptable salts with pharma-ceutically acceptable bases. In these cases, the term "pharma-ceutically acceptable salts" refers to the relatively non-toxic inorganic and organic base addition salts of the 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 acid form with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharma-ceutically acceptable metal cation, with ammonia, or with a pharma-ceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).

[0031] The term "pharmaceutically acceptable cocrystal" refers to a solid-state conformer that does not form symmetric ionic interactions with a small molecule.

[0032] A "therapeutically effective amount" (or "effective amount") of a compound for use in therapy refers to the amount of compound in a preparation that, when administered as part of a desired dosing regimen (to a mammal, preferably a human), relieves the symptoms, ameliorates the condition, or delays the onset of a disease condition, e.g., at a reasonable benefit / risk ratio applicable to any medical treatment, according to clinically acceptable standards for the disease or condition being treated, or for cosmetic purposes.

[0033] The term "prophylactic or therapeutic" treatment is art-recognized and includes administration to a host of one or more of the subject compositions. If it is administered prior to the appearance of clinical symptoms of an undesirable condition (e.g., a disease or other undesirable condition of the host animal), the treatment is prophylactic (i.e., protects the host from the development of the undesirable condition), whereas if it is administered after the appearance of the undesirable condition, the treatment is therapeutic (i.e., aims to reduce, ameliorate, or stabilize an existing undesirable condition or its side effects).

[0034] 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 a human.

[0035] 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 straight, branched, or cyclic aliphatic hydrocarbon groups, and includes saturated and unsaturated aliphatic groups, such as alkyl, alkenyl, or alkynyl groups.

[0036] "Alkyl" refers to a fully saturated, cyclic or acyclic, branched or unbranched carbon chain moiety having the specified number of carbon atoms, or up to 30 carbon atoms if no specification is given. For example, alkyl of 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 of 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 or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C for straight chain). 30 , C3-C for branched chains 30 ), more preferably 20 or less. The alkyl group may be substituted or unsubstituted.

[0037] As used herein, the term "heteroalkyl" refers to an alkyl moiety, as defined above, that contains one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of a carbon atom.

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

[0039] As used herein, the term "hydroxyalkyl" refers to an alkyl group, as defined above, substituted with at least one hydroxyl.

[0040] As used herein, the term "alkylene" refers to an alkyl group having a specified number of carbons, e.g., 2-12 carbon atoms, and including in its longest carbon chain two points of attachment to the remainder of the compound. Non-limiting examples of alkylene groups include methylene-(CH2)-, ethylene-(CH2CH2)-, n-propylene-(CH2CH2CH2)-, isopropylene-(CH2CH(CH3))-, 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.

[0041] "Cycloalkyl" means mono- or bicyclic, or bridged or spirocyclic, or polycyclic saturated carbocyclic rings, each having from 3-12 carbon atoms. Preferred cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3-6 carbons in the ring structure. Cycloalkyl groups can be substituted or unsubstituted.

[0042] As used herein, the term "halocycloalkyl" refers to a cycloalkyl group, as defined above, that is substituted with at least one halogen.

[0043] "Cycloheteroalkyl" refers to a cycloalkyl moiety, as defined above, that includes one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of a carbon atom. Preferred cycloheteroalkyls have from 4-8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4-6 carbon and heteroatoms in the ring structure. Cycloheteroalkyl groups can be substituted or unsubstituted.

[0044] Unless the number of carbons is otherwise specified, "lower alkyl," as used herein, refers to an alkyl group as defined above, but having from 1 to 10 carbons, more preferably 1 to 6 carbon atoms, in its backbone structure, 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 designated herein as alkyl is a lower alkyl.

[0045] "Alkenyl" refers to any cyclic or acyclic, branched or unbranched, unsaturated carbon chain moiety having the number of carbon atoms specified, or up to 26 carbon atoms if no limit is specified, and having one or more double bonds in the moiety. Alkenyls of 6 to 26 carbon atoms are exemplified in their various isomeric forms by hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosoenyl, docosenyl, tricosenyl, and tetracosenyl, where the unsaturated bond(s) can be located at any position in the moiety and can have either the (Z) or (E) configuration around the double bond(s).

[0046] "Alkynyl" refers to a hydrocarbyl moiety within the scope of alkenyl, but which has one or more triple bonds.

[0047] The term "aryl" as used herein includes 3-12 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 are heteroatoms (i.e., heteroaryl). Preferably, the aryl group includes a 5-12 membered ring, more preferably a 6-10 membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings, where two or more carbons are common to two adjacent rings and at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Carbocyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like. Heteroaryl groups include substituted or unsubstituted aromatic 3-12 membered ring structures, more preferably 5-12 membered rings, more preferably 5-10 membered rings, where the ring structures include 1-4 heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, etc. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.

[0048] The terms "halo", "halide", or "halogen" as used herein means halogen, including, but not limited to, for example, 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.

[0049] 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, whose ring structures contain one to four heteroatoms. The heterocycles can be monocyclic, bicyclic, spirocyclic, or polycyclic. Examples of heterocyclyl groups include 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 azetidinone and pyrrolidinone, sultams, sultones, and the like. Heterocycles may be substituted at one or more positions with substituents as described above, 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 moieties, -CF3, -CN, and the like.

[0050] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution results in a stable compound that does not spontaneously undergo transformation, e.g., by rearrangement, cyclization, elimination, and the like, subject to the permissible valences of the substituted atom and substituent. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In one 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 may be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. The substituents can 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 substituted alkyl substituent is C 1-6 Alkyl, C 3-6 In a more preferred embodiment, the substituent of the substituted alkyl is selected from fluoro, carbonyl, cyano, or hydroxyl. Those skilled in the art will appreciate that the substituents may themselves be substituted, if appropriate. Unless specifically stated as "unsubstituted," references to chemical moieties herein are understood to include substituted variants. For example, an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0051] As used herein, the definition of each term, e.g., alkyl, m, n, etc., when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure.

[0052] As used herein, "small molecule" refers to a small organic or inorganic molecule with 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). The 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 2500 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 1500, about 500 to about 1000, about 300 to about 1000 Da, or about 100 to about 250 Da).

[0053] In some embodiments, "small molecule" refers to an organic, inorganic, or organometallic compound that typically has a molecular weight of less than about 1000. In some embodiments, small molecules are organic compounds with sizes on the order of 1 nm. In some embodiments, small molecule drugs of the present invention include oligopeptides and other biomolecules with a molecular weight of less than about 1000.

[0054] An "effective amount" is an amount sufficient to produce a beneficial or desired result. For example, a therapeutic amount is one that achieves a desired therapeutic effect. This amount may 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 may be administered in one or more administrations, applications, or dosages. A therapeutically effective amount of a composition will depend on the composition selected. The composition may be administered one or more times per day to one or more times per week, including once every other day. One of skill in the art will appreciate that certain factors may affect the dosage and timing required to effectively treat a subject, including, but not limited to, the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present. Additionally, treatment of a subject with a therapeutically effective amount of a composition described herein may include a single treatment or a series of treatments.

[0055] The terms "reduce", "reduce", "reduced", "reduction", "reduce", and "inhibit" are all used herein to generally mean a statistically significant reduction compared to a reference. However, for the avoidance of doubt, "reduce", "reduce", or "reduce" or "inhibit" typically means 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%, up to, for example, the complete absence of a given entity or parameter compared to a reference level, or any reduction of 10-99% compared to the absence of a given treatment.

[0056] The terms "increased," "increase" or "enhance," or "activate" are all used herein to mean an increase, generally in a statistically significant amount, and for the avoidance of doubt, the terms "increased," "increase" or "enhance," or "activate" mean an increase of at least 10% compared to a reference level, for example, an increase of 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 a 100% increase compared to a reference level, or any increase between 10-100%, 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 increase, or any increase between 2-fold and 10-fold or more compared to a reference level.

[0057] As used herein, the term "modulate" includes upregulation and downregulation, eg, enhancing or inhibiting a response.

[0058] A "radiopharmaceutical" as defined herein refers to a pharmaceutical containing at least one radiation-emitting radioisotope. Radiopharmaceuticals are routinely used in nuclear medicine for the diagnosis and / or treatment of various diseases. Radiolabeled pharmaceuticals, 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 application of the radiolabeled pharmaceutical. 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 linked to the remainder of the molecule directly or via a linker.

[0059] 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.

[0060] Compounds of the Invention One aspect of the invention is a compound of formula (I) [ka] During the ceremony, L1 is absent or selected from -alkyl-, -cycloalkyl-, and -heteroaryl-CH2-; X1 and X2 are independently selected from -H, alkyl, cycloalkyl, and alkyl-cycloalkyl, with the proviso that X1 and X2 are not both -H; Y1 is optionally substituted aryl; Y2 is selected from optionally substituted pyridonyl, optionally substituted pyrimidinoyl, optionally substituted pyrazinonyl, optionally substituted triazinonyl, and optionally substituted quinazolinonyl; Y3, Y4, Y5, and Y6 are independently selected from -H and a halide; The present invention relates to a compound, or a pharma- ceutically acceptable salt thereof.

[0061] In certain embodiments, the compound has the structure: [ka] has.

[0062] In certain embodiments, one of X1 and X2 is -H, and the other of X1 and X2 is selected from C1-C4 alkyl, cycloalkyl, and alkyl-cycloalkyl.

[0063] In certain embodiments, one of X1 and X2 is -H, and the other of X1 and X2 is selected from C1-C4 alkyl and cycloalkyl. 。

[0064] In certain embodiments, X1 is -H and X2 is -CH3. In other embodiments, X2 is -H and X1 is -CH3. In other embodiments, X1 is -H and X2 is [ka] In other embodiments, X2 is -H and X1 is [ka] It is.

[0065] In certain embodiments, L1 is absent.

[0066] In certain embodiments, L1 is selected from -C1-C4 alkyl-, -cycloalkyl-, and -heteroaryl-CH2-.

[0067] In certain embodiments, L1 is -CH2-. In other embodiments, L1 is [ka] In another embodiment, L1 is [ka] In another embodiment, L1 is selected from: [ka] is selected from.

[0068] In certain embodiments, the compound is [ka] In certain embodiments, the compound has a structure selected from: [ka] The structure is selected from:

[0069] In certain embodiments, Y 1 is unsubstituted aryl.

[0070] In certain embodiments, Y 1 is unsubstituted phenyl.

[0071] In certain embodiments, Y1 is substituted aryl.

[0072] In certain embodiments, Y is [ka] and R1, R2, R3, R4, and R5 are independently selected from -H, halogen, -CN, -CF3, -CHF2, -CF2CH3, -OCF3, -OCHF2, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that at least one of R1, R2, R3, R4, and R5 is not -H.

[0073] In certain embodiments, R1, R2, R3, R4, and R5 are independently -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH3-CF3, -CHF2, -CF2CH3, -C(H)(OH)(CH3), -OCH3, -OCF3, -OCHF2, and [ka] with the proviso that at least one of R1, R2, R3, R4, and R5 is not -H.

[0074] In certain embodiments, R1, R2, R3, R4, and R5 are independently -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH3, and [ka] with the proviso that at least one of R1, R2, R3, R4, and R5 is not -H.

[0075] In certain embodiments, two of R1, R2, R3, R4, and R5 are not -H.

[0076] In certain embodiments, three of R1, R2, R3, R4, and R5 are not -H.

[0077] In certain embodiments, Y is [ka] where R2 is selected from -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, and -CF2CH3.

[0078] In certain embodiments, Y is [ka] wherein R2 and R4 are each independently selected from -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, and -CF2CH3.

[0079] In certain embodiments, Y is [ka] wherein R1 and R4 are each independently selected from -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, and -CF2CH3. In other embodiments, Y1 is [ka] wherein R3 and R4 are each independently selected from -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, and -CF2CH3.

[0080] In certain embodiments, Y is [ka] wherein R1 and R3 are each independently -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, -OCF3, -CF2CH3, and [ka] is selected from.

[0081] In certain embodiments, R1 is -F and R3 is -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, -OCF3, -CF2CH3, and [ka] is selected from.

[0082] In certain embodiments, Y is [ka] It is.

[0083] In certain embodiments, Y is [ka] wherein R1, R3, and R4 are each independently -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, -OCF3, -CF2CH3, -C(H)(OH)(CH3), and [ka] is selected from.

[0084] In certain embodiments, R1 is -F, and R3 and R4 are each independently -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, -OCF3, -CF2CH3, and [ka] is selected from.

[0085] In certain embodiments, R1 is -F, R3 is -Cl or -F, and R4 is -Cl, -Br, -F, -CN, -CH3, -CH2CH3, -OCH3, -OCF3, -CF2CH3, and [ka] is selected from.

[0086] In certain embodiments, the compound is [ka] The structure is selected from:

[0087] In certain embodiments, the compound is [ka] The structure is selected from:

[0088] In certain embodiments, Y2 is selected from unsubstituted pyridonyl, unsubstituted pyrimidinoyl, unsubstituted pyrazinonyl, unsubstituted triazinonyl, and unsubstituted quinazolinonyl.

[0089] In certain embodiments, Y2 is [ka] is selected from.

[0090] In certain embodiments, Y2 is selected from substituted pyridonyl, substituted pyrimidinoyl, substituted pyrazinonyl, substituted triazinonyl, and substituted quinazolinonyl.

[0091] In certain embodiments, Y2 is [ka] and R6 and R7 are independently -H, halogen, -CN, -OH 、 selected from -OCF3, -OCHF2, -NH2, alkyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that at least one of R6 and R7 is not -H; or R6 and R7 together with the carbons to which they are attached form an unsubstituted or substituted fused C5-C7 cycloalkyl.

[0092] In certain embodiments, Y2 is [ka] and R7 and R8 are independently -H, halogen, -CN, -OH 、 -OCF3, -OCHF2, -NH2, alkyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that at least one of R7 and R8 is not -H; or R7 and R8 together with the carbons to which they are attached form an unsubstituted or substituted fused C5-C7 cycloalkyl.

[0093] In certain embodiments, Y2 is [ka] and R6 and R9 are independently -H, halogen, -CN, -OH 、-OCF3, -OCHF2, -NH2, alkyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that at least one of R6 and R9 is not -H.

[0094] In certain embodiments, Y2 is [ka] and R 10 are halogens, -CN, -OH 、 It is selected from -OCF3, -OCHF2, -NH2, alkyl, alkoxy, alkylamino, and cycloalkyl.

[0095] In certain embodiments, Y2 is [ka] and R 11 are halogens, -CN, -OH 、 It is selected from -OCF3, -OCHF2, -NH2, alkyl, alkoxy, alkylamino, and cycloalkyl.

[0096] In certain embodiments, Y2 is [ka] is selected from.

[0097] In certain embodiments, Y2 is N-substituted pyridonyl, N-substituted pyrimidinoyl, N-substituted pyrazinonyl, N-substituted triazinonyl, or N-substituted quinazolinonyl.

[0098] In certain embodiments, Y2 is N-alkyl substituted pyridonyl, N-alkyl substituted pyrimidinoyl, N-alkyl substituted pyrazinonyl, N-alkyl substituted triazinonyl, or N-alkyl substituted quinazolinonyl.

[0099] In certain embodiments, Y2 is [ka] is selected from.

[0100] In certain embodiments, the compound has the structure: [ka] has.

[0101] In certain embodiments, Y3 and Y4 are both -H or both -F. In other embodiments, Y3 is -H and Y4 is -F. In other embodiments, Y4 is -H and Y3 is -F.

[0102] In certain embodiments, Y5 and Y6 are both -H or both -F. In other embodiments, Y5 is -H and Y6 is -F. In other embodiments, Y6 is -H and Y5 is -F.

[0103] In certain embodiments, Y3, Y4, Y5, and Y6 are each -H.

[0104] In certain 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]

[0105] 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, by replacing hydrogen with deuterium or tritium, or carbon with 13 C or 14 Compounds produced by replacing the variable R with C-enriched carbon are within the scope of the invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the invention. For example, 1 In the case of, (C1-C4) alkyl or -O-(C1-C4) alkyl may be suitably deuterated (e.g., -CD3, -OCD3).

[0106] Any of the compounds of the present invention can also be radiolabeled for the preparation of radiopharmaceuticals.

[0107] Treatment methods One aspect of the present invention provides compounds, compositions and methods useful for treating or preventing diseases or disorders associated with abnormal levels of amino acids through modulation of SLC6A19 transport.

[0108] Another aspect of the invention relates to a method of modulating SLC6A19 trafficking in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0109] Another aspect of the 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).

[0110] In some 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).

[0111] In some 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).

[0112] In some embodiments, the compound reduces systemic phenylalanine levels in the subject.

[0113] In some embodiments, the present invention relates to a method of treating or preventing tyrosinemia (Type I, II, or III) in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0114] In some embodiments, the compound reduces systemic glycine levels in the subject.

[0115] In some embodiments, the present invention relates to a method of treating or preventing isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorders, or hyperammonemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0116] In some embodiments of any one of the disclosed methods, the compound modulates SLC6A19 in the subject.

[0117] In some embodiments of any one of the disclosed methods, the compound inhibits SLC6A19 in the subject.

[0118] In some embodiments of any one of the disclosed methods, the compound modulates SLC6A19 trafficking in the subject.

[0119] In some embodiments of any one of the disclosed methods, the compound inhibits SLC6A19 transport in the subject.

[0120] In some embodiments, the compound reduces the systemic level of an amino acid in a subject.

[0121] In some embodiments of any one of the disclosed methods, the subject is a mammal. In some embodiments of any one of the disclosed methods, the mammal is a human.

[0122] In some embodiments of any one of the disclosed methods, the compound of formula (I) is [ka] wherein L1 is absent or selected from -alkyl-, -cycloalkyl-, and -heteroaryl-CH2-; X1 and X2 are independently selected from -H, alkyl, cycloalkyl, and alkyl-cycloalkyl, with the proviso that X1 and X2 are not both -H; Y1 is optionally substituted aryl; Y2 is selected from optionally substituted pyridonyl, optionally substituted pyrimidinoyl, optionally substituted pyrazinonyl, optionally substituted triazinonyl, and optionally substituted quinazolinonyl; Y3, Y4, Y5, and Y6 are independently defined as a compound, or a pharma- ceutically acceptable salt thereof, selected from -H and a halide.

[0123] In some embodiments of any one of the disclosed methods, the compound of formula (I) is [ka] wherein L1 is absent or selected from -alkyl-, -cycloalkyl-, and -heteroaryl-CH2-; X1 and X2 are independently selected from -H, alkyl, cycloalkyl, and alkyl-cycloalkyl, with the proviso that X1 and X2 are not both -H; Y1 is optionally substituted aryl; Y2 is selected from optionally substituted pyridonyl, optionally substituted pyrimidinoyl, optionally substituted pyrazinonyl, optionally substituted triazinonyl, and optionally substituted quinazolinonyl; The compound is defined as a compound, or a pharma- ceutically acceptable salt thereof.

[0124] Pharmaceutical Compositions, Routes of Administration, and Dosages In certain embodiments, the present invention relates to a pharmaceutical composition comprising a compound of the present invention and a pharma- ceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises multiple compounds of the present invention and a pharma- ceutically acceptable carrier.

[0125] In certain embodiments, the pharmaceutical composition of the present invention further comprises at least one additional pharmacologic active agent other than the compound of the present invention. The at least one additional pharmacologic active agent may be an agent useful for treating ischemia-reperfusion injury.

[0126] Pharmaceutical compositions of the present invention can be prepared by combining one or more compounds of the present invention with a pharma- ceutically acceptable carrier and, optionally, one or more additional pharma- ceutical active agents.

[0127] As stated above, "effective amount" refers to any amount that is sufficient to achieve a desired biological effect. By selecting among various active compounds and weighting factors, such as potency, relative bioavailability, patient weight, severity of adverse side effects, and mode of administration, in combination with the teachings provided herein, an effective prophylactic or therapeutic treatment regimen can be designed that is effective in treating a particular subject while not causing substantial undesirable toxicity. The effective amount for any particular application can vary depending on factors such as the disease or condition being treated, the particular compound of the invention being administered, the size of the subject, or the severity of the disease or condition. Those skilled in the art can empirically determine the effective amount of a particular compound of the invention and / or other therapeutic agent without necessitating undue experimentation. A maximum dose, i.e., the highest safe dose according to some medical judgment, may be used. Multiple doses per day may be contemplated to achieve an appropriate systemic level of the compound. An appropriate systemic level may be determined, for example, by measuring the patient's peak or sustained plasma levels of the drug. "Dose" and "administration" are used interchangeably herein.

[0128] In certain embodiments, intravenous administration of the compound may typically be from 0.1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 0.1 mg / kg / day to 2 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 0.5 mg / kg / day to 5 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 1 mg / kg / day to 10 mg / kg / day.

[0129] In general, the daily oral dose of the compound is about 0.01 milligrams / kg per day to 1000 milligrams / kg per day for human subjects. Oral doses ranging from 0.5 to 50 milligrams / kg in one or more administrations per day are expected to provide therapeutic results. The dosage may be appropriately adjusted to achieve the desired local or systemic drug levels, depending on the mode of administration. For example, intravenous administration is expected to be one to several orders of magnitude lower per day. In the event that the response in the subject is inadequate at such doses, even higher doses (or higher effective doses by a different, more localized delivery route) may be used, within the limits of patient tolerance. Multiple doses per day are contemplated to achieve adequate systemic levels of the compound.

[0130] For any compound described herein, therapeutically effective doses can be determined from animal models first. Therapeutically effective doses can also be determined from human data for compounds that have been 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 doses can be adjusted based on the relative bioavailability and potency of the administered compound. It is well within the capabilities of those skilled in the art to adjust the dose to achieve maximum efficacy based on the above methods and other methods known in the art.

[0131] The formulations of the present invention can be administered in pharma- ceutically acceptable solutions, which may routinely contain pharma- ceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.

[0132] For use in treatment, an effective amount of the compound can be administered to a subject by any method that delivers the compound to the desired surface.Administering pharmaceutical compositions can be achieved 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., topically to the eye), inhalation, and topical.

[0133] For intravenous and other parenteral routes of administration, the compounds of the present invention can be formulated as lyophilized preparations, as lyophilized preparations of liposome-entrapped or encapsulated active compounds, as lipid complexes in aqueous suspension, or as salt complexes. Lyophilized preparations are generally reconstituted in a suitable aqueous solution, such as sterile water or saline, immediately prior to administration.

[0134] For oral administration, the compound can be easily formulated by combining the active compound(s) with pharma- ceutically acceptable carriers well known in the art. Such carriers allow the compound 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 optionally milled and, after adding suitable auxiliary agents, processed granule mixtures to obtain tablets or dragee cores as solid excipients. Suitable excipients are in particular fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, e.g., corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or cellulose preparations such as polyvinylpyrrolidone (PVP). If desired, disintegrating agents such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate may be added. Optionally, the oral formulations may also be formulated in saline or buffers, such as EDTA to neutralize internal acid conditions, or may be administered without any carrier.

[0135] Also specifically contemplated are oral dosage forms of the above component or components. The component or components may be chemically modified to effect oral delivery of the derivative. Generally, the contemplated chemical modification is the attachment of at least one moiety to the component molecule itself, which (a) inhibits acid hydrolysis, and (b) allows uptake into the bloodstream from the stomach or intestine. Also desired is an increase in the overall stability of the component or components, and an increase in 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, Hocenberg 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 uses, as indicated above, polyethylene glycol moieties are preferred.

[0136] For the component (or derivative), the site of release may be the stomach, the small intestine (the duodenum, jejunum, or ileum), or the large intestine. Those skilled in the art have formulations available that will not dissolve in the stomach, but will release the material in the duodenum or elsewhere in the intestine. Preferably, the release avoids the deleterious effects of the stomach environment by protection of the compound (or derivative) of the invention or by releasing the biologically active material beyond the stomach environment, such as in the intestine.

[0137] To ensure full gastric resistance, a coating that is impermeable to at least pH 5.0 is essential. Examples of the more common inactive ingredients used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP50, HPMCP55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can be used as mixed films.

[0138] A coating or mixture of coatings can also be used for tablets that are not intended for protection against the stomach. 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), and for liquid forms, a soft gelatin shell can be used. The shell material of cachets can be thick starch or other edible paper. For pills, lozenges, molded tablets, or crushed tablets, wet mashing techniques can be used.

[0139] The therapeutic agent may be included in the formulation as fine multiparticulates in the form of granules or pellets with a particle size of about 1 mm. The formulation of the material for capsule administration may also be as a powder, lightly compressed plugs, or even tablets. The therapeutic agent may be prepared by compression.

[0140] Colorants and flavoring agents may all be included. For example, the compounds (or derivatives) of the present invention may be formulated (e.g., by liposomal or microparticle encapsulation) and then further contained within an edible product, such as a refrigerated beverage, that includes colorants and flavoring agents.

[0141] The volume of the therapeutic agent may be diluted or increased with an inert material. These diluents may include carbohydrates, particularly mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans, and starch. Certain inorganic salts may also be used as fillers, including calcium triphosphate, magnesium carbonate, and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx1500, Emcompress, and Avicell.

[0142] Disintegrants may be included in the formulation of therapeutic agents to make solid dosage forms. Materials used as disintegrants include, but are not limited to, starch, including Explotab, a commercially available disintegrant based on starch. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethylcellulose, natural sponge, and bentonite may all be used. Another form of disintegrant is the insoluble cation exchange resin. Powdered gums may be used as disintegrants and as binders, and these may include powdered gums such as agar, Karaya, or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

[0143] Binders may be used to hold the therapeutic together to form a hard tablet and may include materials from natural products such as acacia, tragacanth, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can both be used in alcoholic solutions to granulate the therapeutic.

[0144] Antifriction agents may be included in the formulation of the therapeutic agent to prevent sticking during the formulation process. Lubricants may be used as a layer between the therapeutic agent and the die wall, and may include, but are not limited to, stearic acid, including magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Soluble lubricants may also be used, such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, Carbowax 4000, and 6000.

[0145] Glidants may be added which may improve the flow properties of the drug during formulation and aid rearrangement during compression. Glidants may include starch, talc, pyrogenic silica, and hydrated silicoaluminate.

[0146] Surfactants may be added as wetting agents to aid in dissolving the therapeutic agent in the aqueous environment. Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate, and dioctyl sodium sulfonate. Cationic detergents may be used and may include benzalkonium chloride and benzethonium chloride. Potential non-ionic detergents that may 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 as a mixture in different ratios in the formulation of the compound or derivative of the present invention.

[0147] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient in a mixture with an injecting agent such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally, a stabilizer. 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. Microparticles formulated for oral administration may also be used. Such microparticles are well defined in the art. All formulations for oral administration must be in dosages suitable for such administration.

[0148] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0149] For local 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 injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as transdermal, transmucosal, oral, or pulmonary administration.

[0150] For administration by inhalation, the compounds for use according to the invention can be conveniently delivered in the form of aerosol spray dispensed from pressurized packs or nebulizers using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve that delivers a metered amount. Capsules and cartridges, for example of gelatin, for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0151] Pulmonary delivery of the compounds disclosed herein (or salts thereof) is also contemplated herein. The compounds are delivered to the lungs of a mammal upon inhalation and cross the pulmonary epithelial lining into the bloodstream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide 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 (α-1-proteinase); Oswein et al., 1990, “Aerosolization of Proteins”, Proceedings of Symposium on Respiratory Drug Delivery, all of which are incorporated by reference. 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 Platz et al., U.S. Patent No. 5,284,656 (granulocyte colony stimulating factor, incorporated by reference). Methods and compositions for pulmonary delivery of drugs for systemic effect are described in U.S. Patent No. 5,451,569, issued September 19, 1995 to Wong et al., incorporated by reference.

[0152] A wide variety of mechanical devices designed for pulmonary delivery of therapeutic products are contemplated for use in the practice of the present invention, including, but not limited to, nebulizers, metered dose inhalers, and dry powder inhalers, all of which are familiar to those skilled in the art.

[0153] Some specific examples of commercially available devices suitable for the practice of 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.

[0154] All such devices require the use of suitable formulations for dispensing the compounds of the present invention. Typically, each formulation is specific to the type of device used and may involve the use of suitable propellant materials in addition to the usual diluents, adjuvants, and / or carriers useful in therapy. Also, the use of liposomes, microcapsules, or microparticles, inclusion complexes, or other types of carriers is contemplated. The chemically modified compounds of the present invention may also be prepared into different formulations depending on the type of chemical modification or the type of device used.

[0155] Formulations suitable for use with either jet or ultrasonic nebulizers typically contain a compound (or derivative) of the invention dissolved in water at a concentration of about 0.1-25 mg of bioactive 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 regulation of osmotic pressure). 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 in forming the aerosol.

[0156] The formulation for use in a metered dose inhaler device generally comprises a finely divided 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 used for this purpose, such as chlorofluorocarbons, hydrochlorofluorocarbons, hydrofluorocarbons, or hydrocarbons, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or combinations thereof. Suitable surfactants include sorbitan trioleate and soybean lecithin. Oleic acid can also be useful as a surfactant.

[0157] Formulations for dispensing from powder inhaler devices include finely divided dry powders containing the compounds (or derivatives) of the invention and may also include bulking agents such as lactose, sorbitol, sucrose, or mannitol in amounts to facilitate dispersion of the powder from the device, e.g., 50-90% by weight of the formulation. The compounds (or derivatives) of the invention should advantageously be prepared in a 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.

[0158] Nasal delivery of the pharmaceutical composition of the present invention is also contemplated.Nasal delivery allows the pharmaceutical composition of the present invention to pass into the bloodstream immediately after administering the therapeutic product to the nose, and does not require the deposition of the product in the lungs.Formulations for nasal delivery include those with dextran or cyclodextran.

[0159] For nasal administration, a useful device is a small hard bottle fitted with a metered spray. In one embodiment, a metered dose is delivered by drawing the pharmaceutical composition solution of the present invention into a chamber of defined volume, which has a hole sized to aerosolize and aerosolize the liquid in the chamber by forming a spray when compressed. The chamber is compressed to administer the pharmaceutical composition of the present invention. In a particular embodiment, the chamber is a piston arrangement. Such devices are commercially available.

[0160] Alternatively, a plastic squeeze bottle is used that has a hole or opening sized to aerosolize the aerosol formulation by forming a spray when squeezed. The opening is usually found at the top of the bottle, which is generally tapered to fit partially into the nasal cavity for efficient administration of the aerosol formulation. Preferably, the nasal inhaler provides a metered amount of the aerosol formulation for administration of a measured dose of the drug.

[0161] Compounds can be formulated for parenteral administration by injection, for example, bolus injection or continuous infusion, when it is desired to deliver them systemically.Injectable preparations can be provided in unit dosage form, for example, in ampoules or multi-dose containers, with added preservatives.Compositions can take the form of suspension, solution, or emulsion in oily or aqueous vehicles, and can contain formulating agents such as suspending agents, stabilizing agents, and / or dispersing agents.

[0162] Pharmaceutical preparations for parenteral administration include aqueous solutions of active compounds in water-soluble form.Additionally, suspensions of active compounds can be prepared as suitable oily injection suspensions.Suitable lipophilic solvents 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 the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran.Optionally, suspensions can also contain suitable stabilizers or agents that increase the solubility of compounds, allowing the preparation of highly concentrated solutions.

[0163] Alternatively, the active compound may be in powder form for constitution with a suitable vehicle, eg, sterile pyrogen-free water, before use.

[0164] 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.

[0165] In addition to the formulations described above, the compounds can also be formulated as depot preparations. Such long-acting preparations can be formulated with 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 a sparingly soluble salt.

[0166] The pharmaceutical compositions may also include 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.

[0167] Suitable liquid or solid pharmaceutical preparation forms are, for example, aqueous or saline solutions for inhalation, microencapsulated, cochleated, coated in microscopic gold particles, contained in liposomes, sprayed, aerosolized, pelleted for implantation in the skin, or dried on a sharp object for rubbing into the skin. Pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or preparations with extended release of active compounds, in which excipients such as disintegrants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners, or solubilizers, as well as additives and / or adjuvants, are usually used as described above. Pharmaceutical compositions are suitable for use in various drug delivery systems. For a brief review of methods for drug delivery, see, for example, Langer R, Science 249:1527-33 (1990).

[0168] The compounds of the present invention and optionally other therapeutic agents may be administered per se (neat) or in the form of a pharma- ceutically acceptable salt or co-crystal. When used in medicine, the salt or co-crystal should be pharma- ceutically acceptable, but pharma-ceutically unacceptable salts or co-crystals may be conveniently used to prepare pharma-ceutically acceptable salts or co-crystals thereof. 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. Such salts may also be prepared as alkali metal or alkaline earth salts, such as sodium, potassium, or calcium salts of the carboxylic acid group.

[0169] 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).

[0170] The pharmaceutical composition of the present invention contains an effective amount of the compound described herein and, optionally, a therapeutic agent contained in a pharma- ceutically acceptable carrier. The term "pharma-ceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents, or encapsulating substances that are suitable for administration to humans or other vertebrates. The term "carrier" refers to a natural or synthetic organic or inorganic component that combines an active ingredient to facilitate application. The components of the pharmaceutical composition can also be mixed with the compound of the present invention and with each other in such a way that there is no interaction that would substantially impair the desired pharmaceutical efficiency.

[0171] Specifically, the therapeutic agent(s), including but not limited to the compounds of the present invention, may be provided in particles. Particles, as used herein, refer to nanoparticles or microparticles (or in some instances larger particles) that may be composed in whole or in 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 within the particle. The particles may be of any order of release kinetics, including zero-order release, first order release, second order release, delayed release, sustained release, immediate release, and any combination thereof. The particles may include, in addition to the therapeutic agent(s), any of these materials routinely used in the pharmaceutical and medical fields, including but not limited to erodible, non-erodible, biodegradable, or non-biodegradable materials, or combinations thereof. The particles may be microcapsules that contain the compounds of the present invention in solution or in a semi-solid state. The particles may be of virtually any shape.

[0172] 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 polymers. The polymer is selected based on the period of desired 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. These include polyhyaluronic acid, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginic acid, 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).

[0173] The therapeutic agent(s) may be included in a controlled release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the means and profile of drug release from the formulation is controlled. It refers to immediate and non-immediate release formulations, which include, but are not limited to, sustained and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in its conventional sense to refer to a drug formulation that provides gradual release of drug over an extended period of time, preferably, but not necessarily, resulting in substantially constant blood drug levels 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 period of delay between administration of the formulation and the release of drug therefrom. "Delayed release" may or may not involve gradual release of drug over an extended period of time, and thus may or may not be "sustained release".

[0174] The use of long-term sustained release implants may be particularly suitable for treating chronic conditions. "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.

[0175] It will be appreciated 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 light of information known to those skilled in the art, and may be made without departing from the scope of the invention or any of its embodiments. Having now described the invention in detail, the 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. EXAMPLES

[0176] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0177] SLC6A19 isoleucine transport assay Cell line generation and maintenance Flp-In™ T-REx™ 293 cell line was purchased from Thermo Fisher Scientific. The line was used to generate stable cell lines inducibly expressing human SLC6A19 with a C-terminal V5 tag and stably expressing human TMEM27 (also known as collectrin) with a C-terminal myc-DDK tag. Stable cell lines were generated by transfecting SLC6A19- and TMEM27-encoding plasmids using standard protocols, followed by antibiotic selection. Stable cells were maintained in DMEM / F12 supplemented with Glutamax, 10% fetal bovine serum, 100U / mL penicillin, 100ug / mL streptomycin, 200ug / mL hygromycin, 10ug / mL blasticidin, and 300ug / mL neomycin (Thermo Fisher).

[0178] Assay: Isoleucine transport assay in 96-well format Stable cell lines were seeded at a density of 35,000 cells per well on day 0 in poly-D-lysine coated 96-well cell culture treated plates. On day 1, expression of SLC6A19 was induced by dispensing tetracycline at a final concentration of 1ug / mL using a Tecan D300e digital dispenser. Transport assays were performed on day 2. Media was removed from the plates using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio) and cells were washed with 175uL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, cells were treated with either 70uL of DMSO, positive control, or compound diluted in Krebs buffer (140mM NaCl, 4.7mM KCl, 2.5mM CaCl2, 1.2mM MgCl2, 11mM HEPES, 10mM glucose, pH 7.4) at room temperature. After 20-60 minutes, 30uL of a 3.3mM solution of 13C6,15N-L-isoleucine (Cambridge Isotope Laboratories) was added. After 20 minutes of incubation with the isoleucine substrate at room temperature, the cells were washed with 175uL of live cell imaging solution using a Blue Washer. The cells were then lysed in 150uL of 15uM D-leucine-d10 (CDN Isotopes) in ultrapure water. The plate was placed on a shaker at 700rpm for a minimum of 40 minutes to facilitate lysis. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the lysates of untreated cells. The plate was returned to the shaker for a minimum of 2 minutes to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000rpm for 5 minutes to pellet and settle the cell debris. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0179] Assay: Isoleucine transport assay in 384-well format On day 0, stable cell lines were seeded into poly-D-lysine coated 384-well cell culture treated plates at a density of 20,000 cells per well using a Viaflo 384-well pipette in medium containing 1ug / mL tetracycline. Transport assays were performed the following day (day 1). Medium was removed from the plates using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio) and cells were washed with 80uL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, cells were treated with either 20uL of DMSO, positive control, or compound diluted in Krebs buffer (140mM NaCl, 4.7mM KCl, 2.5mM CaCl2, 1.2mM MgCl2, 11mM HEPES, 10mM glucose, pH 7.4) using a TECAN liquid holder. After 20-60 min incubation at room temperature, 8.6 uL of a 3.3 mM solution of 13C6,15N-L-isoleucine (Cambridge Isotope Laboratories) was added. After 20 min incubation with isoleucine substrate at room temperature, cells were washed with 80 uL of live cell imaging solution using a Blue Washer. Cells were then lysed in 80 uL of 15 uM D-leucine-d10 (CDN Isotopes) in ultrapure water. Plates were placed on a shaker at 700 rpm for a minimum of 2 hours to facilitate lysis. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing lysates of untreated cells. Plates were returned to the shaker for a minimum of 5 min to ensure proper mixing of the standard curve. Plates were then centrifuged at 4,000 rpm for 10 min to pellet and settle cell debris. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0180] 13C6,15N-L-isoleucine analysis was performed using a RapidFire365-QTOF 6545 (Agilent). Quantitative sample analysis utilizes automated solid phase extraction (HILIC H6 cartridge) prior to mass spectrometric injection. Samples were loaded using 95% acetonitrile, 0.1% formic acid and eluted directly from the cartridge with 5% acetonitrile, 0.1% formic acid for ESI-MS (electrospray ionization) analysis. Quantification of analytes was performed using Agilent Masshunter Quant software from high-resolution full scan data.

[0181] General Procedure General Procedure 1: Synthesis of 3-aryl 5-aminomethylisoxazoles The 3-aryl 5-aminomethylisoxazoles used or referenced below were prepared as appropriate using appropriate modifications of the following procedures and the appropriate starting arenes.

[0182] Synthesis of 3-aryl 5-aminomethylisoxazole A7 [ka] Step 1: Synthesis of compound A2 To a mixture of compound A1 (800 mg, 2.931 mmol) and TEA (888 mg, 8.8 mmol) in DMF (15 mL) was added trimethylsilylacetylene (0.42 mL, 2.931 mmol), CuI (56 mg, 0.293 mmol), and Pd(PPh3)2Cl2 (215 mg, 0.293 mmol). The resulting mixture was stirred at 50 °C under N2 atmosphere for 16 h. The mixture was then diluted with EtOAc (50 mL), filtered, and the filtrate was washed with saturated NH4Cl solution (50 mL x 2), water (50 mL), and brine (50 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with PE:EtOAc=100:0 to 95:5) to give compound A2 (650 mg, yield 91.17%) as a pale yellow solid.

[0183] Step 2: Synthesis of compound A3 To a mixture of compound A2 (650 mg, 2.673 mmol) in MeOH (10 mL) was added K2CO3 (1.85 g, 13.363 mmol). The resulting mixture was stirred at room temperature for 30 min. The mixture was then filtered and the filtrate was concentrated under reduced pressure to give crude compound A3 (350 mg, 76.57% yield) as a yellow oil without further purification. 1H NMR (400 MHz, MeOD) δ 7.47 (t, J = 1.8 Hz, 1H), 7.42 (d, J = 1.8 Hz, 2H), 3.74 (s, 1H).

[0184] Step 3: Synthesis of compound A4 To a mixture of compound A3 (350 mg, 2.046 mmol) and DIEA (532 mg, 4.093 mmol) in THF (12 mL) was slowly added ethyl 2-chloro-2-(hydroxyimino)acetate (465 mg, 3.070 mmol) under N2 atmosphere at 0°C. The resulting mixture was stirred at room temperature for 18 h. The mixture was then diluted with EtOAc (40 mL), washed with water and brine, and the organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 85:15) to give compound A4 (300 mg, 51.24% yield) as a yellow oil. LC / MS (ESI) m / z: 286 (M+H) + .

[0185] Step 4: Synthesis of compound A5 To a solution of compound A4 (300 mg, 1.049 mmol) in anhydrous DCM (12 mL) was added DIBAL (3.2 mL, 1 M in hexanes) at -30°C under N2 atmosphere. The resulting mixture was allowed to warm to room temperature for 2 h. The mixture was then poured into ice-cold NH4Cl solution (30 mL) and extracted with EtOAc (30 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 95:5) to give compound A5 (180 mg, 70.33% yield) as a pale yellow solid. LC / MS (ESI) m / z: 244 (M+H) + .

[0186] Step 5: Synthesis of compound A6 To a solution of compound A5 (180 mg, 0.737 mmol) in toluene (12 mL) was added DPPA (264 mg, 0.959 mmol) and DBU (1.3 mL, 8.112 mmol). The resulting mixture was stirred at room temperature under N2 atmosphere for 18 h. The mixture was diluted with EtOAc (30 mL) and washed with water (40 mL) and brine (40 mL). The organic layer was separated, dried over anhydrous Na2SO4 and concentrated under reduced pressure to give crude compound A6 (197 mg, 99.27% ​​yield) as a colorless oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 269 (M+H). + .

[0187] Step 6: Synthesis of compound A7 To a solution of compound A6 (197 mg, 0.732 mmol) in THF (8 mL) and H2O (2 mL) was added PPh3 (384 mg, 1.464 mmol). The resulting mixture was stirred at room temperature under N2 atmosphere for 18 h. The mixture was then acidified to pH=6 with 1N HCl solution, and the resulting mixture was extracted with methyl tert-butyl ether (40 mL). The aqueous layer was separated and basified to pH=8 with saturated NaHCO3 solution, then extracted twice with EtOAc (30 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give compound A7 (150 mg, 84.29% yield) as a colorless oil. LC / MS(ESI)m / z:243(M+H) + .

[0188] General Procedure 2: Synthesis of 3-aminomethyl 5-arylisoxazoles: The 3-aryl 5-aminomethylisoxazoles used or referenced below were prepared using appropriate modifications of the following procedures and the appropriate starting arenes.

[0189] Synthesis of 3-aminomethyl 5-arylisoxazole B5 [ka] Step 1: Synthesis of compound B2 To a mixture of compound B1 (800 mg, 5.962 mmol) and disodium carbonate (2.53 g, 23.849 mmol) in EtOH (20 mL) was added NH2OH-HCl (1.16 g, 7.751 mmol). The resulting mixture was stirred at 40° C. for 30 min. The mixture was concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (40 mL) and washed with water (40 mL) and brine (40 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give crude compound B2 (880 mg, 98.9% yield) as a white solid, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 150 (M+H) + .

[0190] Step 2: Synthesis of compound 3 To a solution of compound B2 (880 mg, 5.899 mmol) in DMF (15 mL) was added NCS (1.18 g, 8.848 mmol) and the resulting mixture was stirred at 40° C. for 3 h. The mixture was then diluted with saturated NH4Cl solution (40 mL) and extracted with EtOAc (40 mL×2). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give crude compound B3 (1.0 g, 92.32% yield) as a white solid, which was used directly in the next step without further purification.

[0191] Step 3: Synthesis of compound B4 To a mixture of compound 3 (1 g, 5.445 mmol) and DIEA (2.12 g, 16.336 mmol) in anhydrous THF (18 mL), tert-butyl N-(prop-2-yn-1-yl)carbamate (0.85 g, 5.445 mmol) was added dropwise at 0° C. The resulting mixture was stirred at 40° C. for 2 h. The mixture was then concentrated to dryness under reduced pressure, and the residue was purified by column chromatography on silica gel (eluted with PE:EtOAc=100:0 to 5:1) to give compound 4 (1.2 g, 72.88% yield) as a white solid. LC / MS (ESI) m / z: 303 (M+H) + .

[0192] Step 4: Synthesis of compound B5 To a solution of compound B4 (400 mg, 1.323 mmol) in DCM (5 mL) was added TFA (1 mL) dropwise at 0° C. The resulting mixture was stirred at rt for 2 h. The mixture was then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (30 mL) and washed with saturated NaHCO3 solution (60 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give crude compound B5 (220 mg, 82.39% yield) as a colorless oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 203 (M+H) + .

[0193] Synthesis of common intermediate C4: [ka] Step 1: Synthesis of compound C2 To a mixture of compound C1 (310 g, 1547.83 mmol) in DCM (7000 mL) was added AcOH (278.48 g, 4643.49 mmol) and 2,4-dimethoxybenzaldehyde (219.6 mL, 1547-95 mmol) at room temperature under N2 atmosphere. After stirring at 0° C. for 3 h, sodium borohydride (984.3 g, 4643.73 mmol) was added to the above mixture in portions over 2 h. The resulting mixture was stirred at room temperature for 16 h. The mixture was then concentrated to dryness under reduced pressure. The residue was diluted with DCM (5000 mL) and washed with water (5000 mL) and brine (5000 mL). The organic layer was separated, dried over anhydrous Na2SO4 and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give compound C2 (515 g, 94.90% yield) as a pale yellow oil. LC / MS (ESI) m / z: 351 (M+H). + . 1H NMR(400 MHz,MeOD)δ 7.24(d,J=8.3 Hz,1H),6.60(d,J=1.9 Hz,1H),6.54(dd,J=8.3,2.2 Hz,1H),4.13(s,1H),4.01(s,2H),3.88(s,3H),3.81(s,4H),2.93(d,J=11.0 Hz,3H),2.14-2.05(m,1H),1.91(s,2H),1.79-1.72(m,1H),1.46(s,9H).

[0194] Step 2: Synthesis of compound C3 To a mixture of C2 (515 g, 1468.78 mmol), AcOH (1322.5 g, 22042 mmol), and (1-ethoxycyclopropoxy)trimethylsilane (512.0 g, 2937.56 mmol) in EtOH (2000 mL) and THF (8000 mL) was added sodium cyanoborohydride (323.4 g, 5141.76 mmol) in portions over 2 h at 0 °C under N2 atmosphere. The resulting mixture was stirred at 80 °C for 8 h. The mixture was then concentrated to dryness under reduced pressure. The residue was diluted with DCM (5000 mL) and washed with water (5000 mL) and brine (5000 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 96:4) to give compound C3 (406.5 g, 70.8% yield) as a pale yellow oil. LC / MS (ESI) m / z: 391 (M+H). + . 1 H NMR(400 MHz,MeOD)δ 7.27(d,J=8.3 Hz,1H),6.59-6.48(m,2H),4.38(s,1H),4.12(s,2H),4.03-3.96(m,1H),3.83(s,3H),3.80(s,3H),3.00-2.85(m,2H),2.78 -2.63(m,1H),2.50-2.30(m,1H),2.25-2.15(m,1H),1.98(s,1H),1.85-1.69(m,2H),1.43(s,9H),0.68(s,2H)0.53(s,2H).

[0195] Step 3: Synthesis of compound C4 To a solution of C3 (406.5 g, 1039.08 mmol) in DCM solution (4000 mL) was added TFA (1000 mL) dropwise under N2 atmosphere at 0 °C within 2 h. The resulting mixture was stirred at room temperature for 3 h. The mixture was then concentrated to dryness under reduced pressure. The residue was diluted with DCM (2000 mL) and adjusted to pH = 8 by gradually adding 10% Na2CO3 solution. The aqueous layer was then extracted with DCM (1500 mL x 4). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4, and concentrated to give crude C4 (275.0 g, 90.9% yield) as a yellow oil without further purification. LC / MS (ESI) m / z: 291 (M+H) + . 1 H NMR(400 MHz,MeOD)δ 7.18(d,J=8.3 Hz,1H),6.51-6.43(m,2H),3.80-3.76(m,8H),3.19(d,J=8.2 Hz,1H),3.08-3.02(m,1H),2.82-2.72(m,2H),2.65-2.54(m,1H),2.13-2.06(m,1H),2. 03-1.96(m,1H),1.89-1.82(m,1H),1.69-1.47(m,2H),0.53-0.44(m,2H),0.33(d,J=2.6 Hz,2H).

[0196] Synthesis of common intermediate D3 [ka] Step 1: Synthesis of compound D1 To a solution of compound C4 (100 mg, 0.344 mmol) in DMF (8 mL) was added 3,6-dichloropyridazine (190 mg, 1.278 mmol) and K2CO3 (142 mg, 1.033 mmol). The resulting mixture was stirred at 80 °C under N2 atmosphere for 16 h. After cooling, the mixture was diluted with EtOAc (40 mL) and washed with saturated NH4Cl solution (30 mL x 3) and brine (30 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with PE:EtOAc = 100:0 to 2:1) to give compound D1 (120 mg, 86.5% yield) as a colorless oil. LC / MS (ESI) m / z: 403 (M+H) + .

[0197] Step 2: Synthesis of compound D2 To a solution of compound D1 (120 mg, 0.298 mmol) in dioxane (2 mL) and H2O (2 mL) was added DABCO (50 mg, 0.447 mmol) and K2CO3 (123 mg, 0.893 mmol). The resulting mixture was stirred at 80 °C under N2 atmosphere for 16 h. After cooling, the mixture was diluted with EtOAc (40 mL) and washed with water (30 mL) and brine (30 mL). The organic layer was separated, dried over anhydrous Na2SO4, and filtered. The filtrate was evaporated to dryness. The residue was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 20:1) to give compound D2 (90 mg, 78.6% yield) as a colorless oil. LC / MS (ESI) m / z: 385 (M+H) + .

[0198] Step 3: Synthesis of compound D3 Compound D2 (90 mg, 0.234 mmol) was added portionwise to TFA (5 mL) at 0° C. under N2 atmosphere. The resulting mixture was then stirred at 80° C. for 4 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give crude compound D3 (54 mg, 98.4% yield) as a purple oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 235 (M+H).+ .

[0199] Examples 1-11: The compounds in the following table were prepared from the appropriate starting materials, either prepared as described above or commercially available, using appropriate modifications of general procedure 3 and (R)-6-(3-(cyclopropylamino)piperidin-1-yl)pyrimidin-2(1H)-one (D3).

[0200] General Procedure 3 [ka] Synthesis of Compound E2 (Example 6) To a mixture of compound D3 (70 mg, 0.304 mmol) and TEA (150 mg, 1.52 mmol) in anhydrous DCM (6 mL) at 0° C. under N2 atmosphere, a solution of compound E1 (84 mg, 0.365 mmol) and BTC (54 mg, 0.182 mmol) in DCM (5 mL) was added dropwise. The resulting mixture was stirred at room temperature for 2 h and then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (20 mL) and washed with water and brine. The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give E2 (15 mg, 10.1% yield) as a white solid. LC / MS (ESI) m / z: 494 (M+H) + . 1 H NMR(400 MHz,MeOD)δ 8.21-8.12(m,2H),7.90(d,J=1.5 Hz,1H),7.43(d,J=7.5 Hz,1H),6.96(d,J=3.3 Hz,1H),6.18(d,J=7.3 Hz,1H),5.08-4.91(m,1H),4.54-4.42(m,2H),4.18-3.92(m,1H),3.76-3.48(m,1H),3.42-3.32(m,1H),3.04-2.80(m, 1H),2.60-2.48(m,1H),2.32-2.12(m,1H),2.01-1.79(m,2H),1.62-1.46(m,1H),1.02-0.90(m,2H),0.86-0.70(m,2H). [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0201] [ka] Step 1: Synthesis of common intermediate F1 Compound D1 (3.8 g, 9.45 mmol) was added portionwise to TFA (60 mL) at 0° C. under N2 atmosphere. The resulting mixture was then stirred at 80° C. for 5 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give crude compound F1 (2.3 g, 96.6% yield) as a purple oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 253 (M+H). + .

[0202] Examples 12-19: The compounds in the following table were prepared from the appropriate starting materials, either prepared as described above or commercially available, using appropriate modifications of general procedure 4 and (R)-1-(2-chloropyrimidin-4-yl)-N-cyclopropylpiperidin-3-amine (F1).

[0203] General Procedure 4 [ka] Step 1: Synthesis of compound G2 To a mixture of compound G1 (53 mg, 0.264 mmol) and compound F1 (67 mg, 0.264 mmol) in anhydrous DCM (5 mL) was added a solution of triphosgene (55 mg, 0.185 mmol) in TEA (80 mg, 0.792 mmol) and DCM (2 mL) dropwise at -50 °C. The resulting mixture was stirred at 45 °C for 1 h. The mixture was then diluted with H2O (20 mL) and extracted with DCM (20 mL x 2). The combined organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to dryness. The residue was purified by column chromatography silica gel (eluted with DCM:MeOH = 100:1 to 100:5) to give compound G2 (90 mg, 71.67% yield) as a white solid. LC / MS (ESI) m / z: 482 (M+H) + .

[0204] Step 2: Synthesis of Compound G3 (Example 15) To a solution of compound G2 (90 mg, 0.187 mmol) in dioxane (3 mL) and H2O (3 mL), DABCO (84 mg, 0.748 mmol) and K2CO3 (155 mg, 1.122 mmol) were added, and the resulting mixture was stirred at 80 °C for 40 h. The mixture was then diluted with H2O (25 mL) and extracted with EtOAc (25 mL x 2). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by prep-HPLC (Gemini 5 μm C18 250 x 21.2 mm, H2O / MeCN (5-95%) / 0.1% HCOOH) to give compound G3 (12.2 mg, 14.08% yield) as a pale yellow solid. LC / MS (ESI) m / z: 464 (M+H) + . [Table 3-1] [Table 3-2] [Table 3-3]

[0205] Synthesis of common intermediate H3 [ka] Step 1: Synthesis of compound H2 To a mixture of compound H1 (800 mg, 5.534 mmol) and compound C4 (1.6 g, 5.534 mmol) in THF (40 mL) was added Pd(OAc)2 (124 mg, 0.533 mmol) and RuPhos (258 mg, 0.553 mmol), followed by the addition of lithium bis(trimethylsilyl)amide (25 mL, 1 M in THF) in portions at 0° C. under N2 atmosphere. The resulting mixture was stirred at 75° C. for 16 h. The mixture was then cooled and concentrated to dryness under reduced pressure. The residue was diluted with DCM (100 mL) and washed with saturated NH4Cl solution and brine. The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 96:4) to give compound H2 (900 mg, 40.8% yield) as a pale yellow oil. LC / MS (ESI) m / z: 399 (M+H). + . 1 H NMR(400 MHz,MeOD)δ 8.01(d,J=5.7 Hz,1H),6.03(d,J=5.7 Hz,1H),4.68-4.58(m,1H),4.32-4.22(m,1H),3.89(s,3H),3.28-3.19(m,2H),3.04-2.95(m,1H),2.53-2. 46(m,1H),2.17-2.07(m,1H),1.84-1.75(m,1H),1.63-1.54(m,2H),0.73-0.64(m,2H),0.63-0.55(m,2H).

[0206] Step 2: Synthesis of H3 To a solution of compound H2 (100 mg, 0.251 mmol) in MeOH (10 mL) was added concentrated HCl (10 mL, 12N). The resulting mixture was stirred at 90 °C under N2 atmosphere for 48 h. The mixture was then cooled and concentrated to dryness under reduced pressure. The residue was diluted with water (30 mL) and basified to pH = 8 with saturated NaHCO3 solution. The mixture was then extracted with EtOAc (30 mL x 2). The combined organic layers were washed with water and brine, dried over Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 95:5) to give H3 (40 mg, 64.1% yield) as a pale brown oil. LC / MS (ESI) m / z: 235 (M+H) + . 1 H NMR(400 MHz,MeOD)δ 7.61(d,J=6.7 Hz,1H),5.74(d,J=6.7 Hz,1H),4.37-4.31(m,1H),4.01-3.91(m,1H),3.19-3.12(m,1H),3.08-3.01(m,1H),2.88-2.76(m,1H),2.38-2.26(m, 1H),2.12-2.03(m,1H),1.88-1.76(m,1H),1.64-1.55(m,1H),1.53-1.41(m,1H),0.60-0.50(m,2H),0.44-0.32(m,2H).

[0207] Examples 20-28: The compounds in the following table were prepared from the appropriate starting materials, either prepared as described above or commercially available, using appropriate modifications of general procedure 5 and (R)-2-(3-(cyclopropylamino)piperidin-1-yl)pyrimidin-4-ol.

[0208] General Procedure 5 [ka] Step 1: Synthesis of Compound I2 (Example 20) To a mixture of compound I1 (80 mg, 0.384 mmol), TEA (190 mg, 1.92 mmol), and compound H3 (108 mg, 0.462 mmol) in anhydrous DCM (8 mL) at −50° C. under N2 atmosphere, a solution of BTC (68 mg, 0.230 mmol) in DCM (5 mL) was added dropwise. The resulting mixture was stirred at room temperature for 2 h and then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (30 mL) and washed with water and brine. The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give I2 (20 mg, 11.1% yield) as a white solid. [Table 4-1] [Table 4-2] [Table 4-3]

[0209] Synthesis of common intermediate J1 [ka] Step 1: Synthesis of compound E3 Compound H2 (400 mg, 1.004 mmol) was added to TFA (8 mL) and the resulting mixture was stirred at 80 °C under N2 atmosphere for 3 h. After cooling, the mixture was concentrated under reduced pressure to give crude J1 (230 mg, 92.27% yield) as a purple solid without further purification. LC / MS (ESI) m / z: 249 (M+H). + .

[0210] Examples 29-35: The compounds in the following table were prepared from the appropriate starting materials, either prepared as described above or commercially available, using appropriate modifications of general procedure 6 and (R)-N-cyclopropyl-1-(4-methoxypyrimidin-2-yl)piperidin-3-amine (J1).

[0211] General Procedure 6 [ka] Step 1: Synthesis of compound K2 To a mixture of compound K1 (50 mg, 0.305 mmol) and compound J1 (76 mg, 0.305 mmol) in anhydrous DCM (5 mL) was added TEA (92 mg, 0.914 mmol) and triphosgene (46 mg, 0.152 mmol) under N2 atmosphere at -40°C. The resulting mixture was stirred at room temperature under N2 atmosphere for 2 h. The reaction mixture was then diluted with DCM (20 mL) and washed with water (30 mL) and brine (30 mL). The organic layer was separated, dried over anhydrous Na2SO4 and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 95:5) to give compound K2 (40 mg, 29.95% yield) as a pale yellow solid. LC / MS (ESI) m / z: 439 (M+H) + .

[0212] Step 2: Synthesis of Compound K3 (Example 29) To a mixture of compound K2 (40 mg, 0.091 mmol) in EtOH (2 mL) was added concentrated HCl (1 mL) and the resulting mixture was stirred at 80° C. under N2 atmosphere for 2 h. After cooling, the mixture was diluted with water (10 mL) and basified with aqueous NaHCO3 solution in portions to adjust pH=8. The mixture was then extracted twice with DCM (20 mL). The combined organic layers were washed with water (30 mL) and brine (30 mL). The organic layers were separated, dried over anhydrous Na2SO4 and concentrated to dryness. The residue was purified by preparative HPLC to give compound K3 (20 mg, 51.65% yield) as a white solid. LC / MS (ESI) m / z: 425 (M+H) + . 1H NMR(400 MHz,MeOD)δ 7.57(d,J=11.0 Hz,1H),7.43(d,J=9.6 Hz,1H),7.36(d,J=7.1 Hz,1H),7.05(t,J=5.9 Hz,1H),5.74(d,J=6.4 Hz,1H),4.46(d,J=5.4 Hz,2H),4.35-4.22(m,2H),3.70(t,J=11.7 Hz,1H),3.38-3.30(m,1H),2.91-2.80(m,1H),2.62-2.54(m,1H),2.50(s,3H),2.32-2. 15(m,1H),2.00-1.81(m,2H),1.67-1.51(m,1H),1.02-0.90(m,2H),0.86-0.74(m,2H).

Table 5-1

Table 5-2

Table 5-3

[0213] Synthesis of common intermediate L3

change

[0214] Step 2: Synthesis of compound L3 Compound L2 (68 mg, 0.175 mmol) was added portionwise to TFA (3 mL) at 0° C. under N2 atmosphere. The resulting mixture was then stirred at 80° C. for 4 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give crude compound L3 (40.32 mg, 98.61% yield) as a purple oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 236 (M+H). + .

[0215] Examples 36-42: The compounds in the following table were prepared from the appropriate starting materials, either prepared as described above or commercially available, using appropriate modifications of general procedure 7 and (R)-3-(3-(cyclopropylamino)piperidin-1-yl)-1,2,4-triazin-5-ol (L3).

[0216] General Procedure 7 Synthesis of compound L5 To a solution of compound L3 (41 mg, 0.171 mmol) in anhydrous DCM (3 mL) at 0° C. under N2 atmosphere, L4 (35 mg, 0.171 mmol) and TEA (52 mg, 0.513 mmol) were added. The mixture was stirred at room temperature for 1 h and then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (20 mL) and washed with water and brine. The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give L5 (13 mg, 16.58% yield) as a white solid. LC / MS (ESI) m / z: 435 (M+H) + . 1 H NMR(400 MHz,MeOD)δ 7.35(s,1H),7.25(d,J=8.0 Hz,1H),7.14(d,J=9.8 Hz,1H),6.93(s,1H),4.41-4.18(m,4H),3.72-3.61(m,1H),3.41(t,J=12.2 Hz,1H),2.94-2.82(m,1H),2.56(s,1H),2.32(s,3H),2.27-2.18(m,1H),1 .98-1.83(m,2H),1.66-1.54(m,1H),1.02-0.90(m,2H),0.85-0.73(m,2H). [Table 6-1] [Table 6-2] [Table 6-3]

[0217] Synthesis of M4 (Example 44) [ka] Step 1: Synthesis of compound M1 To a mixture of compound C4 (100 mg, 0.344 mmol) and Cs2CO3 (337 mg, 1.03 mmol) in anhydrous DMF (8 mL) was added 6-amino-2-chloro-1H-pyrimidin-4-one (101 mg, 0.689 mmol). The resulting mixture was stirred at 120 °C for 3 h under microwave. The mixture was then cooled, diluted with EtOAc (30 mL), washed with saturated NH4Cl solution (30 mL x 3) and brine (20 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 90:10) to give compound M1 (112 mg, 81.4% yield) as a pale yellow solid. LC / MS (ESI) m / z: 400 (M+H) + .

[0218] Step 2: Synthesis of compound M2 Compound M1 (112.0 g, 0.280 mmol) was added portionwise to TFA (6 mL) at 0° C. under N2 atmosphere. The resulting mixture was then stirred at 80° C. for 4 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give crude compound M2 (69 mg, 98.72% yield) as a purple oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 250 (M+H). + .

[0219] Step 3: Synthesis of compound M4 To a mixture of compound M2 (69 mg, 0.277 mmol), TEA (84 mg, 0.830 mmol), and M3 (44.17 mg, 0.277 mmol) in anhydrous DCM (10 mL) was added triphosgene (41.06 mg, 0.138 mmol) at −50° C. under N2 atmosphere. The resulting mixture was allowed to warm slowly to room temperature and stirred for an additional 2 h. The mixture was then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (20 mL) and washed with water (20 mL) and brine (20 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give M4 (15.0 mg, 12.46% yield) as a white solid. [ka]

[0220] Synthesis of N5 (Example 45) Step 1: Synthesis of compound N1 To a mixture of compound C4 (1 g, 3.448 mmol) and DIEA (1.33 g, 10.345 mmol) in anhydrous MeCN (20 mL) was added 2,4,6-trichloro-1,3,5-triazine (635 mg, 3.448 mmol) dropwise at 0° C. under N2 atmosphere. The resulting mixture was stirred at room temperature for 1 h. The mixture was concentrated to dryness under reduced pressure. The residue was diluted with EtOAc (60 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with PE:EA=100:0 to 90:10) to give compound L1 (1.3 g, 86.6% yield) as a pale yellow solid. LC / MS (ESI) m / z: 437 (M+H) + .

[0221] Step 2: Synthesis of compound N2 To a mixture of compound N1 (1.3 g, 2.294 mmol) in MeCN (15 mL) was added aqueous NaOH (15 mL, 1N). The resulting mixture was stirred at room temperature for 12 h. The mixture was then diluted with EtOAc (50 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 90:10) to give compound N2 (802 mg, 64.5% yield) as a colorless oil. LC / MS (ESI) m / z: 419 (M+H) + .

[0222] Step 3: Synthesis of compound N3 To a mixture of compound N2 (802 mg, 1.914 mmol) in MeCN (10 mL) was added aqueous NaOH (10 mL, 6N). The resulting mixture was stirred at 100° C. for 12 h. After cooling, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 90:10) to give compound N3 (370 mg, 48.2% yield) as a colorless oil. LC / MS (ESI) m / z: 402 (M+H) + .

[0223] Step 4: Synthesis of compound N4 Compound N3 (370 mg, 0.92 mmol) was added portionwise to TFA (10 mL) at 0° C. under N2 atmosphere. The resulting mixture was then stirred at 80° C. for 4 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give crude compound N4 (200 mg, 86.9% yield) as a purple oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 252 (M+H). + .

[0224] Step 5: Synthesis of compound N5 To a mixture of M4 (80 mg, 0.503 mmol), TEA (254 mg, 2.52 mmol), and compound N4 (152 mg, 0.604 mmol) in anhydrous DCM (10 mL) was added a solution of BTC (90 mg, 0.302 mmol) in DCM (5 mL) dropwise at −50° C. under N2 atmosphere. The resulting mixture was stirred at room temperature for 2 h and then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (20 mL) and washed with water (20 mL) and brine (20 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH=100:0 to 95:5) to give compound L5 (45 mg, 20.3% yield) as a white solid.

[0225] Examples 43-44: The compounds in the following table were prepared from (R)-N-cyclopropyl-N-(3,4-dimethoxybenzyl)piperidin-3-amine (C4) according to the procedure described above. [Table 7]

[0226] Synthesis of common intermediate O4: Step 1: Synthesis of compound O2 [ka] To a solution of tert-butyl N-[(3R)-3-piperidyl]carbamate (10 g, 49.93 mmol) O1 and DIPEA (7.10 g, 54.92 mmol, 9.57 mL) in DCM solution (500 mL) was added 2-nitrobenzenesulfonyl chloride (11.62 g, 52.43 mmol) as a solid in small portions. The reaction mixture was allowed to stir at ambient temperature for 15 minutes, then TFA (56.93 g, 499.31 mmol, 38.47 mL) was added slowly. The reaction mixture was stirred at ambient temperature for an additional 2 hours, then concentrated under reduced pressure by rotary evaporation. The crude product was then resuspended in 500 mL of DCM and to this solution was added DIPEA (11.29 g, 87.38 mmol, 15.22 mL), 2,4-dimethoxybenzaldehyde (7.88 g, 47.43 mmol), and sodium triacetoxyborohydride (26.46 g, 124.83 mmol). The reaction mixture was allowed to stir overnight at ambient temperature. The reaction mixture was then washed with 1 M NaOH (500 mL), then the organic layer was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure by rotary evaporation. The crude residue was purified by flash column chromatography (DCM:MeOH, 100:0 to 96:4) to provide (3R)-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (7.94 g, 18.23 mmol, 36.52% yield) O2.

[0227] Step 1: Synthesis of compound O3 To a solution of (3R)-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (7.93 g, 18.21 mmol)O2 in THF (240 mL) and EtOH (120 mL) was added 1-ethoxy-1-trimethylsiloxycyclopropane (7.94 g, 45.52 mmol, 9.15 mL), sodium cyanoborohydride (4.01 g, 63.73 mmol), and acetic acid (16.40 g, 273.14 mmol, 15.62 mL). The reaction mixture was stirred overnight at 80° C., after which it was cooled to rt and concentrated under reduced pressure by rotary evaporation. The residue was then dissolved in ethyl acetate (250 mL), washed with 1M NaOH (250 mL), brine (250 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure by rotary evaporation to provide (3R)-N-cyclopropyl-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (8.58 g, 18.04 mmol, 99.08% yield) O3, which was used without further purification.

[0228] Step 1: Synthesis of compound O4 (3R)-N-cyclopropyl-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (8.58 g, 18.04 mmol)O3 was dissolved in TFA (100 mL) and Et3SiH (10 mL) and heated to 80° C. After 4 hours, an additional portion of triethylsilane (7.28 g, 62.61 mmol, 10 mL) was added to suppress the formation of the dimethoxytryl cation. The reaction mixture was then allowed to stir overnight at room temperature. The reaction mixture was then concentrated under reduced pressure by rotary evaporation and the crude residue was dissolved in 200 mL of ethyl acetate. The organic layer was washed with 3M NaOH (approximately 200 mL), brine (approximately 200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure by rotary evaporation. The crude residue was then dissolved in 100 mL of diethyl ether and HCl (2.0 M in diethyl ether, 9.02 mL) was added slowly dropwise. The product was filtered from solution to provide (3R)-N-cyclopropyl-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (5.5 g, 15.20 mmol, 84.25% yield, HCl salt) O4 as a tan solid.

[0229] Examples 45-48: The compounds in the following table were prepared from the appropriate starting materials, either prepared as described above or commercially available, using appropriate modifications of general procedure 8 and ((3R)-N-cyclopropyl-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (O4).

[0230] General procedure 8: [ka] Step 1: Synthesis of compound P2 To a vigorously stirred solution of P1 (183.28 mg, 506.52 μmol) in water (3 mL) and DCM (3 mL) was added sodium bicarbonate (255.32 mg, 3.04 mmol). The solution was cooled to 0° C. and then triphosgene (82.67 mg, 278.59 μmol) was added in one portion. The solution was stirred for 1 h and O4 (183.28 mg, 506.52 μmol) was added in one portion. The solution was then diluted with water and DCM. The phases were separated and the aqueous phase was extracted twice more with DCM. The combined organic phase was washed with brine, dried over MgSO4, and concentrated to a residue. The residue was purified by silica column chromatography (30-100% EtOAc / heptane). P2 (179.5 mg, 296.95 μmol, 58.63% yield) was isolated as a colorless oil.

[0231] Step 2: Synthesis of compound P3 To a solution of P2 (180 mg, 297.78 μmol) in DMF (1 mL) was added potassium carbonate (82.31 mg, 595.56 μmol) and thiophenol (49.21 mg, 446.67 μmol, 45.78 μL). The resulting solution was stirred at room temperature for 2 h, then diluted with water (4 mL) and acidified with HCl (12 M, 248.15 μL). The resulting suspension was extracted with toluene (3×5 mL) and the organic phase was discarded. The aqueous phase was basified with NaOH (6 M, 1.49 mL) and then extracted with EtOAc (4×5 mL). The combined organic phase was washed with brine, dried over MgSO4, and concentrated to give P3 (127 mg, 302.88 μmol, 100%) as a yellow oil, which was used without further purification.

[0232] Step 1: Synthesis of Compound P4 (Example 46) To a solution of P3 (25 mg, 59.62 umol) in DMF (250 uL) was added DIPEA (21 uL, 120 umol) and 4-tert-butoxy-2-chloro-pyrimidine (14.5 mg, 77 umol). The solution was heated at 60° C. for 1 h, at which point LCMS indicated consumption of starting material. The solution was concentrated in vacuo to remove DMF, then TFA (500 uL) was added. The solution was stirred at room temperature for 1 h, at which point LCMS indicated cleavage of the t-butyl protection. The solution was concentrated again, then taken up in methanol and purified by preparative reverse phase HPLC. [Table 8-1] [Table 8-2]

[0233] Examples 49-60: The compounds in the following table were either prepared as described above, commercially available, or prepared from the appropriate starting materials according to general procedure 8, by using appropriate modifications of general procedure 9 and ((3R)-N-cyclopropyl-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (O4).

[0234] Synthesis of Compound P5 (Example 59) [ka] General procedure 9: To a solution of P3 (15 mg, 35.77 umol) in DMF (250 uL) was added DIPEA (13 uL, 72 umol) and 4-tert-butoxy-2-chloro-pyrimidine (10 mg, 47 umol). The solution was heated at 60° C. for 1 h, at which point LCMS indicated consumption of starting material. The solution was concentrated in vacuo to remove DMF, then TFA (500 uL) was added. The solution was stirred at room temperature for 1 h, at which point LCMS indicated cleavage of the t-butyl protection. The solution was concentrated again, then taken up in methanol and purified by preparative reverse phase HPLC. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4]

[0235] Examples 61-62: The compounds in the following table were prepared from intermediates such as Q1, which were themselves prepared according to General Procedure 8, and appropriately substituted heteroarenes following appropriate modification of General Procedure 10 shown below. [ka] General procedure: To a solution of Q1 (35 mg, 107 umol) in DMF (1 mL) was added DIPEA (28 uL, 161 umol) and the appropriately substituted heteroarene (118 umol). The solution was heated at 60° C. for 1 h, at which point LCMS indicated consumption of starting material. The product was then directly purified by preparative reverse-phase HPLC. [Table 10]

[0236] Additional compounds listed in the table below were prepared by adapting the experimental procedures listed above. [Table 11-1] [Table 11-2] [Table 11-3]

[0237] General procedure for synthesis of 11-71 [ka] Step 1 To a mixture of R1 (100 mg, 0.458 mmol) and DIPEA (119 mg, 0.916 mmol) in anhydrous MeCN (15 mL) was added R2 (103 mg, 0.550 mmol) or an appropriately substituted heteroarene. The resulting mixture was stirred at 90 °C under N2 atmosphere for 16 h. The mixture was then diluted with EtOAc (30 mL) and washed with water (30 mL) and brine (30 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with PE:EtOAc = 10:1 to 3:1) to give R3 (102 mg, 60.4% yield) as a colorless oil. LC / MS (ESI) m / z: 369 (M+H) + .

[0238] Step 2 To a solution of R3 (102 mg, 0.276 mmol) in DCM (4 mL) was added TFA (1 mL) at 0° C. and the reaction was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give crude R4 (TFA salt) as a colorless oil, which was used for the next step without any further purification. LC / MS (ESI) m / z: 213 (M+H). + .

[0239] Step 3 To a solution of R4 (0.276 mmol) in anhydrous DCM (8 mL) were added 2,4-dimethoxybenzaldehyde (49 mg, 0.291 mmol) and AcOH (67 mg, 1.104 mmol) at room temperature. After the mixture was stirred at room temperature for 1.5 h, NaBH(OAc)3 (117.3 mg, 0.554 mmol) was added at 0 °C. The resulting mixture was then stirred at room temperature overnight. After the reaction was completed, the mixture was quenched with saturated aqueous NaHCO3 (20 mL) to adjust pH = 8 and extracted with DCM (20 mL x 4). The combined organic layer was washed with brine (25 mL), dried over anhydrous Na2SO4, and concentrated to dryness under reduced pressure. The residue was purified by chromatography on silica gel (DCM:MeOH = 50:1 to 15:1) to give R5 (60 mg, 60.1% yield) as a colorless oil. LC / MS(ESI)m / z:363(M+H) + .

[0240] Step 4 To a mixture of R5 (60 mg, 0.166 mmol) and AcOH (99.4 mg, 1.66 mmol) in THF (12 mL) and EtOH (6 mL) was added (1-ethoxycyclopropoxy)trimethylsilane (86.6 mg, 0.500 mmol), followed by NaBH3CN (31.2 mg, 0.500 mmol). The resulting mixture was stirred at 80 °C under N2 atmosphere for 4 h. The mixture was then cooled to room temperature and concentrated to dryness. The residue was diluted with DCM (15 mL), poured into saturated NaHCO3 (50 mL) and adjusted to pH = 8. The mixture was then extracted with DCM (20 mL x 2) and the combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated to dryness. The crude product was purified on silica gel by column chromatography (DCM:MeOH=100:1 to 20:1) to give R6 (62 mg, 92.9% yield) as a colorless oil. LC / MS(ESI) m / z: 403(M+H) + .

[0241] Step 5 R6 (62.0 mg, 0.154 mmol) was added portionwise to TFA (5 mL) at 0° C. under N2 atmosphere, and the resulting mixture was stirred at 80° C. for 4 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give crude R7 (TFA salt) as a purple oil, which was used for the next step without any further purification. LC / MS (ESI) m / z: 253 (M+H). + .

[0242] Step 6 To a mixture of R7 (0.154 mmol) and TEA (47 mg, 0.462 mmol) in anhydrous DCM (6 mL) was added a solution of isocyanate (0.169 mmol) in anhydrous DCM (2 mL) dropwise over 5 min at 0 °C under N2 atmosphere, and the resulting mixture was then stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in EtOAc (10 mL), washed with water (15 mL) and brine (15 mL). The organic layer was separated, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by prep-HPLC (Gemini 5 μm C18 250 × 21.2 mm, H2O / MeCN (5-95%) / 0.1% NH3·H2O) to give 73 (25 mg, 33.29% yield) as a white solid. LC / MS (ESI) m / z: 488 (M+H) + . 1 H NMR(400 MHz,MeOD)δ 7.65(s,1H),7.45(t,J=8.6 Hz,1H),7.13-6.93(m,3H),5.80(d,J=6.6 Hz,1H),4.67-4.53(m,2H),4.45(d,J=5.4 Hz,2H),4.21-4.15(m,1H),3.82-3.69(m,1H),2.89-2.79(m,1H),2.62 -2.53(m,1H),2.43-2.32(m,2H),1.01-0.92(m,2H),0.86-0.77(m,2H).

[0243] Incorporation by Reference All U.S. patents and U.S. and PCT patent application publications cited herein are hereby incorporated by reference.

[0244] 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 which equivalents are intended to be encompassed by the following claims.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, L １ is absent or is selected from -alkyl-, -cycloalkyl-, and -heteroaryl-CH ２ - is selected from, X １ and X ２ is independently selected from —H, alkyl, cycloalkyl, and alkyl-cycloalkyl, with the proviso that X １ and X ２ However, both do not become -H, Y １ is optionally substituted aryl; Y ２ is selected from optionally substituted pyridonyl, optionally substituted pyrimidinoyl, optionally substituted pyrazinonyl, optionally substituted triazinonyl, and optionally substituted quinazolinonyl; Y ３ , Y ４ , Y ５ , and Y ６ are independently selected from —H and halides.

2. X １ and X ２ is —H, and X １ and X ２ The other is C １ -C ４ selected from alkyl, cycloalkyl, and alkyl-cycloalkyl; X １ and X ２ is —H, and X １ and X ２ The other is C １ -C ４ selected from alkyl and cycloalkyl; X １ is -H, and X ２ But -CH ３ is; X ２ is -H, and X １ But -CH ３ is; X １ is -H, and X ２ but, 【Chemistry 2】 is; or X ２ is -H, and X １ but, 【Transformation 3】 2. The compound of claim 1, wherein:

3. L １ But it doesn't exist; L １ But, -C １ -C ４ Alkyl-, -cycloalkyl-, and -heteroaryl-CH ２ - selected from; L １ But -CH ２ - is; L １ but, 【Chemistry 4】 is; L １ but, 【Transformation 5】 or L １ but, 【Transformation 6】 2. The compound of claim 1, selected from: 【Request Item 4】 【Chemistry 7】 10. The compound of claim 1, having a structure selected from:

5. Y １ is unsubstituted aryl; Y １ is unsubstituted phenyl; Y １ is substituted aryl; or Y １ but, 【Transformation 8】 where: R １ , R ２ , R ３ , R ４ , and R ５ are independently —H, halogen, —CN, or —CF ３ , -CHF ２ , -CF ２ CH ３ , -OCF ３ , -OCHF ２ , alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that R １ , R ２ , R ３ , R ４ , and R ５ at least one of is not —H; R １ , R ２ , R ３ , R ４ , and R ５ are independently —H, —F, —Cl, —Br, —CN, or —CH ３ , -CH ２ CH ３ , -CF ３ , -CHF ２ , -CF ２ CH ３ , -C(H)(OH)(CH ３ ), -OCH ３ , -OCF ３ , -OCHF ２ , and 【Chemistry 9】 is selected from the group consisting of １ , R ２ , R ３ , R ４ , and R ５ at least one of is not —H; or R １ , R ２ , R ３ , R ４ , and R ５ are independently —H, —F, —Cl, —Br, —CN, or —CH ３ , -CH ２ CH ３ , and 【Chemistry 10】 is selected from the group consisting of １ , R ２ , R ３ , R ４ , and R ５ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein at least one of: is not —H.

6. R １ , R ２ , R ３ , R ４ , and R ５ two of which are not —H; or R １ , R ２ , R ３ , R ４ , and R ５ 6. The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein three of are not —H.

7. Y １ but, 【Chemistry 11】 wherein R ２ is -Cl, -Br, -F, -CN, -CH ３ , -CH ２ CH ３ , -OCH ３ , and -CF ２ CH ３ Selected from: Y １ but, 【Chemistry 12】 wherein R ２ and R ４ each independently represents —Cl, —Br, —F, —CN, or —CH ３ , -CH ２ CH ３ , -OCH ３ , and -CF ２ CH ３ Selected from: Y １ but, 【Chemistry 13】 wherein R １ and R ４ each independently represents —Cl, —Br, —F, —CN, or —CH ３ , -CH ２ CH ３ , -OCH ３ , and -CF ２ CH ３ Selected from: Y １ but, 【Chemistry 14】 wherein R ３ and R ４ each independently represents —Cl, —Br, —F, —CN, or —CH ３ , -CH ２ CH ３ , -OCH ３ , and -CF ２ CH ３ Selected from: Y １ but, 【Chemistry 15】 wherein R １ and R ３ each independently represents —Cl, —Br, —F, —CN, or —CH ３ , -CH ２ CH ３ , -OCH ３ , -OCF ３ , -CF ２ CH ３ , and 【Chemistry 16】 or R １ is -F and R ３ is -Cl, -Br, -F, -CN, -CH ３ , -CH ２ CH ３ , -OCH ３ , -OCF ３ , -CF ２ CH ３ , and 【Chemistry 17】 Selected from: Y １ but, [Chemistry 18] is; or Y １ but, 【Chemistry 19】 wherein R １ , R ３ , and R ４ each independently represents —Cl, —Br, —F, —CN, or —CH ３ , -CH ２ CH ３ , -OCH ３ , -OCF ３ , -CF ２ CH ３ , -C(H)(OH)(CH ３ ), and 【Chemistry 20】 R １ is -F and R ３ and R ４ each independently represents —Cl, —Br, —F, —CN, or —CH ３ , -CH ２ CH ３ , -OCH ３ , -OCF ３ , -CF ２ CH ３ , and 【Chemistry 21】 or R １ is -F and R ３ is —Cl or —F, and R ４ is -Cl, -Br, -F, -CN, -CH ３ , -CH ２ CH ３ , -OCH ３ , -OCF ３ , -CF ２ CH ３ , and 【Chemistry 22】 2. The compound of claim 1, selected from: 【Request Item 8】 【Chemistry 23】 10. The compound of claim 1, having a structure selected from:

9. Y ２ is selected from unsubstituted pyridonyl, unsubstituted pyrimidinoyl, unsubstituted pyrazinonyl, unsubstituted triazinonyl, and unsubstituted quinazolinonyl; Y ２ but, 【Chemistry 24】 Selected from: Y ２ is selected from substituted pyridonyl, substituted pyrimidinoyl, substituted pyrazinonyl, substituted triazinonyl, and substituted quinazolinonyl; Y ２ but, 【Chemistry 25】 and R ６ and R ７ are independently —H, halogen, —CN, or —OH 、 -OCF ３ , -OCHF ２ , -NH ２ , alkyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that R ６ and R ７ is not —H, or R ６ and R ７ together with the carbon to which they are attached to form an unsubstituted or substituted fused C ５ -C ７ Forming a cycloalkyl; Y ２ but, 【Chemistry 26】 and R ７ and R ８ are independently —H, halogen, —CN, or —OH 、 -OCF ３ , -OCHF ２ , -NH ２ , alkyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that R ７ and R ８ is not —H, or R ７ and R ８ together with the carbon to which they are attached to form an unsubstituted or substituted fused C ５ -C ７ Forming a cycloalkyl; Y ２ but, 【Chemistry 27】 and R ６ and R ９ are independently —H, halogen, —CN, or —OH 、 -OCF ３ , -OCHF ２ , -NH ２ , alkyl, alkoxy, alkylamino, and cycloalkyl, with the proviso that R ６ and R ９ at least one of is not —H; Y ２ but, 【Chemistry 28】 and R １０ But halogen, -CN, -OH 、 -OCF ３ , -OCHF ２ , -NH ２ , alkyl, alkoxy, alkylamino, and cycloalkyl; Y ２ but, 【Chemistry 29】 and R １１ But halogen, -CN, -OH 、 -OCF ３ , -OCHF ２ , -NH ２ , alkyl, alkoxy, alkylamino, and cycloalkyl; Y ２ but, 【Transformation 30】 Selected from: Y ２ is N-substituted pyridonyl, N-substituted pyrimidinoyl, N-substituted pyrazinonyl, N-substituted triazinonyl, or N-substituted quinazolinonyl; Y ２ is N-alkyl-substituted pyridonyl, N-alkyl-substituted pyrimidinoyl, N-alkyl-substituted pyrazinonyl, N-alkyl-substituted triazinonyl, or N-alkyl-substituted quinazolinonyl; or Y ２ but, 【Chemistry 31】 2. The compound of claim 1, selected from:

10. Y ３ , Y ４ , Y ５ , and Y ６ are each —H; Y ３ and Y ４ are both -H or both -F; Y ３ is -H, and Y ４ is -F; Y ４ is -H, and Y ３ is -F; Y ５ and Y ６ are both -H or both -F; Y ５ is -H, and Y ６ is -F; or Y ６ is -H, and Y ５ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

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

12. A pharmaceutical composition comprising the compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

13. 12. A drug for treating or preventing a disease or disorder associated with a genetic defect in phenylalanine hydroxylase; phenylketonuria; hyperphenylalaninemia; tyrosinemia (type I, II, or III); nonketotic hyperglycinemia; isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorder, or hyperammonemia in a subject; or diabetes, chronic kidney disease, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, metabolic syndrome, obesity-related disorder, or neurodevelopmental disorder and autism spectrum disorder, the drug comprising a compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof.

14. systemic phenylalanine levels in the subject are reduced; The systemic tyrosine level in the subject is reduced; systemic glycine levels in the subject are reduced; or The agent of claim 13, wherein SLC6A19 function in the subject is inhibited.

Images