Treatment of PKU using mammalian SLC6A19 function correcting agents

Abstract

Disclosed are compounds, compositions and methods useful for treating or preventing diseases or conditions associated with abnormal levels of amino acids by modulating SLC6A19 transport.

Description

[0001] Related Applications

[0002] This application claims the priority benefit of U.S. Provisional Patent Application No. 63 / 406,443, filed on September 14, 2022. Background Art

[0003] Phenylketonuria (PKU) is a congenital metabolic error caused by mutations in phenylalanine hydroxylase (PAH), the enzyme responsible for metabolizing phenylalanine. PKU is an autosomal recessive metabolic disorder in which phenylalanine cannot be properly metabolized and results in abnormally high plasma phenylalanine levels. Individuals with PKU have abnormally high levels of phenylalanine in their blood, which, if left untreated, can lead to irreversible nerve damage, resulting in a series of complications such as intellectual disability, epilepsy, neurodevelopmental and behavioral disorders. PKU is difficult to treat because the level of phenylalanine in the blood is directly related to diet. Patients must adhere to a strict diet for life, which affects all aspects of the patient's life. The current standard of care is enzyme cofactor and enzyme replacement therapies, but these therapies are not effective for all patients and carry a risk of potential adverse events.

[0004] The enzyme responsible for metabolizing phenylalanine and thus maintaining phenylalanine homeostasis is phenylalanine hydroxylase (PAH). Loss-of-function (LOF) mutations at the PAH gene, known to be located on chromosome 12q23.2, cause most forms of PKU. These LOF mutations that cause PKU can be diagnosed as classic PKU (the most severe form), while "mild PKU" or "hyperphe" is a less severe form. In addition to PAH, mutations in other enzymes that affect phenylalanine metabolism, such as dihydropteridine reductase (DHPR), the enzyme responsible for synthesizing the cofactor required for PAH activity, can also lead to elevated phenylalanine levels. In addition to diet, blood amino acid levels, including phenylalanine levels, are also regulated by SLC6A19. SCL6A19 is located in the proximal tubules of the kidney and is responsible for reabsorbing amino acids back into the blood. Summary of the Invention

[0005] 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 by modulating SLC6A19 transport.

[0006] Accordingly, compounds having the structure of formula (I) are provided herein:

[0007]

[0008] Wherein:

[0009] n is 0 or 1;

[0010] L1 is absent or selected from -NH-, -N(CH 3 )-, -O- and -CH 2 -;

[0011] L 2 is -alkyl-

[0012] L 3 is -(5-membered heteroaryl)-;

[0013] X 1 is -C(R 1 )(R 2 )(R 3 );

[0014] X 2 is optionally substituted aryl or heteroaryl;

[0015] X 3 is selected from -H, alkyl and haloalkyl;

[0016] R 1 is selected from -H, halo, hydroxy, amido, amino, alkylamino and aminoalkyl; and

[0017] R 2 and R 3 are each independently selected from -H and alkyl; or R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl;

[0018] provided that the compound is not selected from

[0019]

[0020] or a pharmaceutically acceptable salt thereof.

[0021] 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, which comprises administering to the subject an effective amount of a compound of formula (I).

[0022] Another aspect of the invention relates to a method of treating or preventing phenylketonuria, hyperphenylalaninemia, tyrosinemia, non-ketotic hyperglycinemia, isovaleric academia, methylmalonic academia, propionic academia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorder or hyperammonemia in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of formula (I).

[0023] Another aspect of the invention relates to a method of modulating SLC6A19 transport in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of formula (I).

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the following describes suitable methods and materials. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0025] Other features, objects, and advantages of the invention will be apparent from the detailed description and the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 is a table summarizing the isoleucine transport data of exemplary compounds of the invention. A = IC 50 < 500 nM; B = IC 50 500 nM - 1500 nM; C = IC 50 > 1500 nM - 5000 nM; D = IC 50 > 5000 nM - 10000 nM; and E = IC 50 > 10000 nM. DETAILED DESCRIPTION

[0027] Definitions

[0028] For convenience, certain terms employed in this specification, the examples, and the appended claims are collected here before further description of the invention. These definitions should be read in light of the remainder of the disclosure and should be as understood by one of ordinary skill in the art. Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0029] To more readily understand the invention, certain terms and phrases are defined below and throughout the specification.

[0030] As used herein, the article "a / an" refers to one or more than one (i.e., at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0031] As used herein in the specification and claims, the phrase "and / or" shall be understood to mean "either or both" of the elements so joined (i.e., elements that coexist in some cases and separate in other cases). Multiple elements listed with "and / or" shall be understood in the same way, i.e., "one or more" of the elements so joined. In addition to the elements specifically identified by the "and / or" clause, other elements may optionally exist, whether related or unrelated to those specifically identified. Thus, as a non-limiting example, when used in conjunction with open-ended language such as "comprising", a reference to "A and / or B" may in one embodiment refer to only A (optionally including elements other than B); in another embodiment refer to only B (optionally including elements other than A); in yet another embodiment refer to both A and B (optionally including other elements); and so on.

[0032] As used herein in the specification and claims, "or" shall be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted inclusively, i.e., including at least one of a number of elements or a list of elements, but also including more than one element, as well as optionally other items not listed. Only terms that clearly indicate the contrary meaning, such as "only one" or "exactly one", or when used in the claims, "consisting of", shall refer to including exactly one of a number of elements or a list of elements. In general, the term "or" as used herein shall be interpreted as indicating exclusive alternatives (i.e., "one or the other but not both") only when followed by an exclusive term such as "either", "one of which", "only one of which", or "exactly one of which". "Consisting essentially of", when used in the claims, shall have the ordinary meaning as used in the field of patent law.

[0033] As used herein in the specification and claims, the phrase "at least one," when referring to a list of one or more elements, is to 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 each specific element listed in the list of elements, and also not excluding any combinations of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, whether related or unrelated to the specifically identified elements. Thus, as a non-limiting example, in one embodiment, "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 to at least one (optionally including more than one) A without B (and optionally including elements other than B); in another embodiment, to at least one (optionally including more than one) B without A (and optionally including elements other than A); in yet another embodiment, to at least one (optionally including more than one) A, and at least one (optionally including more than one) B (and optionally including other elements); and so on.

[0034] It should also be understood that, unless the context otherwise requires, in any method claimed herein that includes more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited.

[0035] In the claims, as well as 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 to be open-ended, i.e., meaning including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" are to be considered closed or semi-closed transitional phrases, as set forth in section 2111.03 of the United States Patent Office Manual of Patent Examining Procedures.

[0036] 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 all such compounds that fall within the scope of the present invention, including cis and trans isomers, R- and S-enantiomers, diastereoisomers, (D)-isomers, (L)-isomers, their racemic mixtures, and other mixtures thereof. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and their mixtures are intended to be included within the present invention.

[0037] "Geometric isomers" means isomers that differ in the orientation of the substituting atoms relative to a carbon-carbon double bond, relative to a cycloalkyl ring, or relative to a bridged bicyclic system. The atoms (other than H) on each side of the carbon-carbon double bond can be in the E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents on the same side) configuration. "R", "S", "S*", "R*", "E", "Z", "cis" and "trans" denote configurations relative to the core molecule. Some of the compounds disclosed herein may exist in "atropisomeric" form or as "atropisomers". Atropisomers are stereoisomers resulting from restricted rotation about a single bond, where the steric strain barrier to rotation is high enough to permit the isolation of conformational isomers. The compounds of the present invention can be prepared as individual isomers by isomer-specific synthesis or can be resolved from a mixture of isomers. Conventional resolution techniques include forming salts of the free bases of each isomer of an isomer pair with an optically active acid (followed by fractional crystallization and regeneration of the free base); forming salts of the acid form of each isomer of an isomer pair with an optically active amine (followed by fractional crystallization and regeneration of the free acid); forming esters or amides of each isomer of an isomer pair with an optically pure acid, amine, or alcohol (followed by chromatographic separation and removal of the chiral auxiliary); or resolving an isomer mixture of the starting material or the final product using various well-known chromatographic methods.

[0038] For example, if a particular enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or by derivation with a chiral auxiliary, where the resulting mixture of diastereoisomers is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, in cases where the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), diastereoisomeric salts can be formed with a suitable optically active acid or base, and the diastereoisomers so formed can then be resolved by fractional crystallization or chromatographic means well known in the art, and the pure enantiomer can then be recovered.

[0039] The purity percentage by mole fraction is the ratio of the number of moles of the enantiomer (or diastereomer) to the number of moles of the enantiomer (or diastereomer) plus the number of moles of its optical isomer. When the stereochemistry of the disclosed compound is named or depicted by structure, the purity of the named or depicted stereoisomer relative to other stereoisomers is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction. When a single enantiomer is named or depicted by structure, the purity of the depicted or named enantiomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction. When a single diastereomer is named or depicted by structure, the purity of the depicted or named diastereomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction.

[0040] When the disclosed compound is named or depicted by a structure that does not indicate stereochemistry and the compound has at least one chiral center, it should be understood that the name or structure encompasses the enantiomers of the compound without the corresponding optical isomers, the racemic mixture of the compound, or a mixture in which one enantiomer is enriched relative to its corresponding optical isomer. When the disclosed compound is named or depicted by a structure that does not indicate stereochemistry and has two or more chiral centers, it should be understood that the name or structure encompasses diastereomers without other diastereomers, many diastereomers without other diastereomer pairs, mixtures of diastereomers, mixtures of diastereomer pairs, diastereomer mixtures in which one diastereomer is enriched relative to one or more other diastereomers, or diastereomer mixtures in which one or more diastereomers are enriched relative to other diastereomers. The present invention encompasses all these forms.

[0041] The structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by replacing hydrogen with deuterium or tritium, or carbon with 13 C- or 14 C-enriched carbon are within the scope of the present invention.

[0042] As used herein, the term "prodrug" encompasses compounds that are converted to a therapeutically active agent under physiological conditions. Common methods for preparing prodrugs include hydrolysis under physiological conditions to expose a selected moiety of the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of the host animal.

[0043] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject chemical from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients in the formulation, not injurious to the patient, and substantially pyrogen-free. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; (4) tragacanth powder; (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 soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. In certain embodiments, the pharmaceutical compositions of the invention are pyrogen-free, i.e., they do not cause a significant increase in body temperature when administered to a patient.

[0044] The term "pharmaceutically acceptable salt" refers to relatively non-toxic inorganic and organic acid addition salts of one or more of the said compounds. These salts can be prepared in situ during the final isolation and purification of one or more of the said compounds, or by separately reacting one or more of the purified compounds in their free base form with a suitable organic or inorganic acid and isolating the salt so formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napsylate, mesylate, glucoheptonate, lactobionate and lauryl sulfonate, etc. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66: 1-19.)

[0045] In other cases, the compounds useful in the methods of the present invention may contain one or more acidic functional groups and can therefore form pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these cases, the term "pharmaceutically acceptable salt" refers to the relatively non-toxic inorganic and organic base addition salts of one or more compounds. These salts can also be prepared in situ during the final isolation and purification of one or more of the said compounds, or can be prepared by separately reacting one or more purified compounds in their free acid form with a suitable base, such as a hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali metal salts or alkaline earth metal salts include lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts and aluminum salts, etc. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc. (see, e.g., Berge et al., supra).

[0046] The term "pharmaceutically acceptable cocrystal" refers to a solid coform that does not form formal ionic interactions with small molecules.

[0047] The "therapeutically effective amount" (or "effective amount") of a compound for use in therapy refers to the amount of the compound in a formulation that, when administered as part of a desired dosing regimen (to a mammal, preferably a human), alleviates symptoms, ameliorates the disorder or slows the onset of the disease disorder according to clinically acceptable criteria for the disorder or condition to be treated or for cosmetic purposes, e.g., with a reasonable benefit / risk ratio applicable to any pharmaceutical treatment.

[0048] The term "prophylactic or therapeutic" treatment is well recognized in the art and includes administering one or more of the subject compositions to a host. If the treatment is administered prior to the clinical manifestation of an unwanted disorder (e.g., a disease or other unwanted condition in a host animal), the treatment is prophylactic (i.e., it protects the host from developing the unwanted disorder), while if the treatment is administered after the manifestation of the unwanted disorder, the treatment is therapeutic (i.e., it is intended to alleviate, ameliorate or stabilize the existing unwanted disorder or its side effects).

[0049] The term "patient" or "subject" refers to a mammal in need of a specific treatment. In certain embodiments, the patient is a primate, canine, feline or equine. In certain embodiments, the patient is a human.

[0050] Aliphatic chains include the alkyl, alkenyl and alkynyl classes defined below. Straight-chain aliphatic chains are limited to the unbranched carbon chain portion. As used herein, the term "aliphatic group" refers to a straight-chain, branched-chain or cyclic aliphatic hydrocarbon group and includes saturated and unsaturated aliphatic groups, such as alkyl, alkenyl or alkynyl.

[0051] "Alkyl" refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having a specified number of carbon atoms or up to 30 carbon atoms if not specified. For example, alkyl having 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl and those that are positional isomers of these moieties. Alkyl having 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, and tetracosyl. In certain embodiments, a straight-chain or branched-chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C 1 -C 30 for a straight chain and C 3 -C 30 for a branched chain), and more preferably 20 or fewer carbon atoms. An alkyl may be substituted or unsubstituted.

[0052] 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 carbon atoms.

[0053] As used herein, the term "haloalkyl" refers to an alkyl as defined above that is substituted with at least one halogen.

[0054] As used herein, the term "hydroxyalkyl" refers to an alkyl as defined above that is substituted with at least one hydroxy group.

[0055] As used herein, the term "alkylene" refers to an alkyl having a specified number of carbon atoms, e.g., 2 to 12 carbon atoms, that contains two attachment points to the remainder of the compound on the longest carbon chain. Non-limiting examples of alkylene include methylene -(CH 2 )-, ethylene -(CH 2 CH 2 ), n-propylene -(CH 2 CH 2 CH 2 ), isopropylidene -(CH 2 CH(CH 3 ))-, etc. An alkylene may be a cyclic or acyclic, branched or unbranched carbon chain moiety and may optionally be substituted with one or more substituents.

[0056] "Cycloalkyl" means a monocyclic or bicyclic or bridged or spiro or polycyclic saturated carbocyclic ring each having 3 to 12 carbon atoms. Preferred cycloalkyls have 3 to 10 carbon atoms in their ring structure, and more preferably 3 to 6 carbon atoms in the ring structure. A cycloalkyl may be substituted or unsubstituted.

[0057] As used herein, the term "halocycloalkyl" refers to a cycloalkyl group substituted with at least one halogen as defined above.

[0058] "Heterocycloalkyl" refers to a cycloalkyl moiety as defined above that contains one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms. Preferred heterocycloalkyls have 4-8 carbon atoms and heteroatoms in their ring structure, and more preferably 4-6 carbon atoms and heteroatoms in their ring structure. Heterocycloalkyls can be substituted or unsubstituted.

[0059] Unless otherwise specified in terms of carbon number, "lower alkyl" as used herein means an alkyl group as defined above, but having from one to ten carbon atoms, more preferably from one to six carbon atoms, in the main chain 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 the application, preferred alkyl groups are lower alkyl groups. In certain embodiments, substituents designated as alkyl groups herein are lower alkyl groups.

[0060] "Alkenyl" refers to any cyclic or acyclic, branched or unbranched unsaturated carbon chain moiety having the specified number of carbon atoms, or having up to 26 carbon atoms if no limitation on the number of carbon atoms is specified; and having one or more double bonds located within the moiety. Examples of alkenyls having 6 to 26 carbon atoms are hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosenyl, docosenyl, tricosenyl, and tetracosenyl in various isomeric forms, where one or more unsaturated bonds can be located at any position within the moiety and can have the (Z) or (E) configuration around one or more double bonds.

[0061] "Alkynyl" refers to a hydrocarbon moiety within the range of alkenyls, but having one or more triple bonds located within the moiety.

[0062] As used herein, the term "aryl" includes 3- to 12-membered substituted or unsubstituted monocyclic aromatic groups, wherein each atom of the ring is carbon (i.e., carbocyclic aryl) or one or more atoms are heteroatoms (i.e., heteroaryl). Preferably, aryl includes 5- to 12-membered rings, more preferably 6- to 10-membered rings. The term "aryl" also includes polycyclic systems having two or more rings, wherein two or more carbons are common to two adjacent rings, and wherein at least one of the rings is aromatic. For example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic. Carbocyclic aryl includes benzene, naphthalene, phenanthrene, phenol, aniline, etc. Heteroaryl includes substituted or unsubstituted aromatic 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10-membered rings, the ring structures of which include one to four heteroatoms. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, etc. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.

[0063] As used herein, the terms "halo", "halogen group" or "halogen" mean halogen and include, for example, but not limited to fluorine, chlorine, bromine, iodine, etc. in radioactive and non-radioactive forms. In a preferred embodiment, halo is selected from the group consisting of fluorine, chlorine, and bromine.

[0064] The term "heterocyclic group" or "heterocyclic moiety" refers to a 3- to 12-membered ring structure, more preferably 5- to 12-membered rings, more preferably 5- to 10-membered rings, the ring structures of which contain 1 to 4 heteroatoms. The heterocycle can be monocyclic, bicyclic, spirocyclic, or polycyclic. Heterocyclic groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxazine, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinoxaline, 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, lactone, lactam (such as azetidinone and pyrrolidone), sultam, sultone, etc. The heterocycle can be substituted at one or more positions by such substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, mercapto, imino, amido, phosphate / ester group, phosphonate / ester group, phosphinate / ester group, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CF 3 , -CN, etc.

[0065] The term "substituted" refers to a moiety having a substituent that replaces hydrogen on one or more carbons of the backbone. It is understood that "substitution" or "substituted with" includes the implicit condition that such substitution is in accordance with the allowed valences of the substituting atoms and the substituents, and that the substitution results in a stable compound, e.g., one that does not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" contemplates all allowed substituents of organic compounds. In a broad aspect, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Allowed substituents can be one or more substituents and can be the same or different for a suitable organic compound. For the purposes of this invention, heteroatoms such as nitrogen can have hydrogen substituents and / or any allowed substituents of the organic compounds described herein that satisfy the valence of the heteroatom. Substituents can include any of the substituents described herein, such as halogen, hydroxy, carbonyl (such as carboxy, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thiocarboxylate), alkoxy, phosphoryl, phosphate / ester, phosphonate / ester, phosphinate / ester, amino, amido, amidine, imine, cyano, nitro, azido, mercapto, alkylthio, sulfate / ester, sulfonate / ester, sulfamoyl, sulfonamido, sulfonyl, heterocyclic, aralkyl, or aromatic or heteroaromatic moieties. In a preferred embodiment, the substituents on the substituted alkyl are selected from C 1-6 alkyl, C 3-6 cycloalkyl, halogen, carbonyl, cyano or hydroxy. In a more preferred embodiment, the substituents on the substituted alkyl are selected from fluorine, carbonyl, cyano or hydroxy. Those skilled in the art will understand that the substituents themselves can be substituted where appropriate. Unless specifically stated as "unsubstituted", references to chemical moieties herein should be understood to include substituted variants. For example, references to "aryl" or moieties implicitly include substituted and unsubstituted variants.

[0066] As used herein, the definition of each expression (e.g., alkyl, m, n, etc.), when it appears more than once in any structure, is intended to be independent of its definition elsewhere in the same structure.

[0067] As used herein, "small molecule" refers to a small organic or inorganic molecule having a molecular weight of less than about 3,000 Daltons. Generally, the 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 Da to about 3,000 Da, about 100 Da to about 2500 Da, about 100 Da to about 2,000 Da, about 100 Da to about 1,750 Da, about 100 Da to about 1,500 Da, about 100 Da to about 1,250 Da, about 100 Da to about 1,000 Da, about 100 Da to about 750 Da, about 100 Da to about 500 Da, about 200 Da to about 1500 Da, about 500 Da to about 1000 Da, about 300 Da to about 1000 Da, or about 100 Da to about 250 Da).

[0068] In some embodiments, "small molecule" refers to an organic, inorganic, or organometallic compound that generally has a molecular weight of less than about 1000. In some embodiments, the small molecule is an organic compound of a size of about 1 nm. In some embodiments, the small molecule drugs of the present invention encompass oligopeptides and other biomolecules having a molecular weight of less than about 1000.

[0069] "Effective amount" is an amount sufficient to produce a beneficial or desired result. For example, a therapeutic amount is an amount that achieves the desired therapeutic effect. This amount can be the same as or different from a prophylactically effective amount, which is an amount necessary to prevent the onset of a disease or disease symptoms. The effective amount can be administered in one or more administrations, applications, or dosages. The therapeutically effective amount of the composition depends on the composition selected. The composition can be administered once or more times per day to once or more times per week; including once every other day. Those skilled in the art will understand that certain factors may affect the dosage and schedule required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the overall health and / or age of the subject, and the presence of other diseases. In addition, the treatment of a subject with a therapeutically effective amount of the composition described herein can include a single treatment or a series of treatments.

[0070] The terms "decrease", "reduce", "reduced", "reduction", "decrease" and "inhibit" are generally all used herein to mean a statistically significant decrease in amount relative to a reference. However, for the avoidance of doubt, "reduce", "reduction" or "decrease" or "inhibit" generally means a decrease of at least 10% compared to a reference level, and may include, for example, a decrease 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 and including (for example) the complete absence of a given entity or parameter, or a decrease between 10 - 99% compared to the situation in the absence of a given treatment.

[0071] The terms "increased", "increase" or "enhance" or "activate" are generally all used herein to mean an increase in a statistically significant amount; for the avoidance of any 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 and including a 100% increase, or any increase between 10% - 100%, or an increase of 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 compared to a reference level, or any increase between 2-fold and 10-fold or more.

[0072] As used herein, the term "modulate" includes upregulation and downregulation, such as enhancing or inhibiting a response.

[0073] "Radiopharmaceutical", as defined herein, refers to a pharmaceutical agent containing at least one radioactive isotope that emits radiation. Radiopharmaceuticals are routinely used in nuclear medicine for the diagnosis and / or therapy of various diseases. A radiolabeled pharmaceutical agent, such as a radiolabeled antibody, contains a radioactive isotope (RI) that serves as a radiation source. As contemplated herein, the term "radioactive isotope" includes metallic and non-metallic radioactive isotopes. The radioactive isotope is selected based on the medical application of the radiolabeled pharmaceutical agent. When the radioactive isotope is a metallic radioactive isotope, a chelating agent is typically used to bind the metallic radioactive isotope to the rest of the molecule. When the radioactive isotope is a non-metallic radioactive isotope, the non-metallic radioactive isotope is typically directly or via a linker connected to the rest of the molecule.

[0074] For the purposes of the present invention, chemical elements are identified according to the CAS version of the Periodic Table of the Elements, Handbook of Chemistry and Physics, 67th Edition, 1986 - 87, inside front cover.

[0075] Compounds of the invention

[0076] One aspect of the present invention relates to compounds of formula (I):

[0077]

[0078] Wherein:

[0079] n is 0 or 1;

[0080] L 1 Is absent or selected from -NH-, -N(CH 3 )-, -O- and -CH 2 -;

[0081] L 2 Is -alkyl-

[0082] L 3 Is -(5 - membered heteroaryl)-;

[0083] X 1 Is -C(R 1 )(R 2 )(R 3 );

[0084] X 2 Is optionally substituted aryl or heteroaryl;

[0085] X 3 Is selected from -H, alkyl and haloalkyl;

[0086] R 1Selected from -H, halo, hydroxy, amido, amino, alkylamino, and aminoalkyl; and

[0087] R 2 and R 3 are each independently selected from -H and alkyl; or R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl;

[0088] provided that the compound is not selected from

[0089]

[0090] or a pharmaceutically acceptable salt thereof.

[0091] In certain embodiments, R 2 and R 3 are each independently selected from -H and alkyl; or R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl that is not cyclopropyl or cycloheteroalkyl.

[0092] In certain embodiments, at least one of R 1 , R 2 and R 3 is not -H. In other embodiments, at least two of R 1 , R 2 and R 3 are not -H. In other embodiments, each of R 1 , R 2 and R 3 is not -H.

[0093] In certain embodiments, the compound has the following structure:

[0094]

[0095] In certain embodiments, L 1 is selected from -NH- and -N(CH 3 )-.

[0096] In certain embodiments, L 2 is selected from -CH 2 -, -CH 2 CH 2 - and -CH 2 CH 2 CH 2 -.

[0097] In certain embodiments, R 1Selected from -H, -F, -OH, -NH 2 , -CH 2 NH 2 , -N(H)(CH 3 ), -N(CH 3 ). 2 and -C(O)NH 2 .

[0098] In certain embodiments, R 1 is -H. In other embodiments, R 1 is -NH 2 .

[0099] In certain embodiments, R 2 and R 3 are each -H. In other embodiments, R 2 and R 3 are each -CH 3 .

[0100] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl.

[0101] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are attached form an unsubstituted cyclopropyl or cyclobutyl.

[0102] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are connected form an optionally substituted cycloheteroalkyl.

[0103] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are attached form an unsubstituted azetidinyl, pyrrolidinyl, piperidinyl or lactam.

[0104] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are attached form a substituted azetidinyl, pyrrolidinyl, piperidinyl or lactam.

[0105] In certain embodiments, the azetidinyl, pyrrolidinyl, piperidinyl or lactam is N-alkyl or N-acetyl substituted.

[0106] In certain embodiments, X 1 is selected from

[0107] In certain embodiments, X 1 is selected from

[0108] In certain embodiments, L 3 is a triazolyl, oxazolyl or oxadiazolyl group.

[0109] In certain embodiments, -L 3 -X 2 is selected from In other embodiments, -L 3 -X 2 is

[0110] In certain embodiments, X 2 is an unsubstituted aryl.

[0111] In certain embodiments, the unsubstituted aryl is an unsubstituted phenyl.

[0112] In certain embodiments, X 2 is a substituted aryl.

[0113] In certain embodiments, the substituted aryl is a substituted phenyl.

[0114] In certain embodiments, X 2 is and

[0115] R 4 、R 5 、R 6 、R 7 and R 8 are independently selected from -H, halogen, -CN, -CF 3 、-CHF 2 、-OCF 3 、-OCHF 2 、alkyl, alkenyl, alkynyl and cycloalkyl; provided that at least one of R 4 、R 5 、R 6 、R 7 and R 8 is not -H.

[0116] In certain embodiments, R 4 、R 5 、R 6 、R 7 and R 8 are independently selected from -H, -Cl, -Br, -F, -CN, -CF 3 、-OCF 3 、-CH3 and cyclopropyl; provided that R 4 , R 5 , R 6 , R 7 and R 8 in at least one is not -H.

[0117] In certain embodiments, X 2 is and R 4 is selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

[0118] In certain embodiments, X 2 is and R 5 is selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

[0119] In certain embodiments, X 2 is and R 6 is selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

[0120] In certain embodiments, X 2 is and R 5 and R 6 are independently selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

[0121] In certain embodiments, X 2 is and R 4 and R 6 are independently selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

[0122] In certain embodiments, X 2 is and R 5 , R 6 and R7 Independently selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

[0123] In certain embodiments, X 2 is selected from:

[0124] In certain embodiments, X 2 is selected from:

[0125] In certain embodiments, X 3 is -H, -CH 3 or -CF 3 . In other embodiments, X 3 is -CF 3 .

[0126] In certain embodiments, n is 1. In other embodiments, n is 2.

[0127] In certain embodiments, the compound has the following structure:

[0128]

[0129] In certain embodiments, the compound has the following structure:

[0130]

[0131] In certain embodiments, the compound has the following structure:

[0132]

[0133] In certain embodiments, the compound has the following structure:

[0134]

[0135] In certain embodiments, the compound has the following structure:

[0136] In certain embodiments, the compound has the following structure:

[0137]

[0138] In some embodiments, the compound is selected from Table 1 below:

[0139] Table 1.

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152] In some embodiments, the compound is an atropisomer. Additionally, unless otherwise specified, the structures depicted herein are also intended to include such compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by replacing hydrogen with deuterium or tritium, or carbon with 13 C- or 14 C-enriched carbon are within the scope of the present invention. Such compounds can be used, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the present invention. For example, with respect to the variable R 1 , (C 1 -C 4 )alkyl or -O-(C 1 -C 4 )alkyl can be suitably deuterated (e.g., -CD 3 , -OCD 3 ).

[0153] Any compound of the present invention can also be radiolabeled for use in the preparation of radiopharmaceuticals.

[0154] Therapeutic methods

[0155] 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 by modulating SLC6A19 transport.

[0156] Another aspect of the invention relates to a method of modulating SLC6A19 transport in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I).

[0157] 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, which comprises administering to the subject an effective amount of a compound of formula (I).

[0158] In certain embodiments, the invention relates to a method of treating or preventing phenylketonuria in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of formula (I).

[0159] In certain embodiments, the invention relates to a method of treating or preventing hyperphenylalaninemia in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of formula (I).

[0160] In some embodiments, the compound reduces the systemic phenylalanine level in the subject.

[0161] In certain embodiments, the invention relates to a method of treating or preventing tyrosinemia (type I, II or III) in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of formula (I).

[0162] In certain embodiments, the compound reduces the systemic glycine level in the subject.

[0163] In certain embodiments, the invention relates to a method of treating or preventing isovaleric academia, methylmalonic academia, propionic academia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorder or hyperammonemia in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of formula (I).

[0164] In certain embodiments of any of the disclosed methods, the compound modulates SLC6A19 in the subject.

[0165] In certain embodiments of any of the disclosed methods, the compound inhibits SLC6A19 in the subject.

[0166] In certain embodiments of any of the disclosed methods, the compound modulates SLC6A19 transport in the subject.

[0167] In certain embodiments of any of the disclosed methods, the compound inhibits SLC6A19 transport in the subject.

[0168] In certain embodiments, the compound reduces the systemic amino acid level in the subject.

[0169] In certain embodiments of any of the disclosed methods, the subject is a mammal. In certain embodiments of any of the disclosed methods, the mammal is a human.

[0170] In certain embodiments of any of the disclosed methods, the compound of formula (I) is defined as:

[0171]

[0172] Wherein:

[0173] n is 0 or 1;

[0174] L 1 is absent or selected from -NH-, -N(CH 3 )-, -O- and -CH 2 -;

[0175] L 2 is -alkyl-

[0176] L 3 is -(5-membered heteroaryl)-;

[0177] X 1 is -C(R 1 )(R 2 )(R 3 );

[0178] X 2 is optionally substituted aryl or heteroaryl;

[0179] X 3 is selected from -H, alkyl and haloalkyl;

[0180] R 1 is selected from -H, halo, hydroxy, amido, amino, alkylamino and aminoalkyl; and

[0181] R 2 and R 3 are each independently selected from -H and alkyl; or R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl;

[0182] or a pharmaceutically acceptable salt thereof.

[0183] In certain embodiments of any of the disclosed methods, the compound is selected from the structure of any of the compounds listed in Table 1.

[0184] In certain embodiments of any of the disclosed methods, the compound is selected from:

[0185]

[0186] In certain embodiments of any of the disclosed methods, the compound is selected from:

[0187]

[0188] wherein:

[0189] n is 0 or 1;

[0190] L 1 is absent or selected from -NH-, -N(CH 3 )-, -O-, and -CH 2 -;

[0191] L 2 is -alkyl-

[0192] L 3 is -(5-membered heteroaryl)-;

[0193] X 1 is -C(R 1 )(R 2 )(R 3 );

[0194] X 2 is optionally substituted aryl or heteroaryl;

[0195] X 3 is selected from -H, alkyl, and haloalkyl;

[0196] R 1 is selected from -H, halo, hydroxy, amido, amino, alkylamino, and aminoalkyl; and

[0197] R 2 and R 3 are each independently selected from -H and alkyl; or R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted cycloalkyl or cycloheteroalkyl;

[0198] provided that the compound is not selected from

[0199]

[0200] or a pharmaceutically acceptable salt thereof.

[0201] Pharmaceutical compositions, routes of administration and dosing

[0202] In certain embodiments, the present invention relates to pharmaceutical compositions comprising a compound of the present invention and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises a plurality of compounds of the present invention and a pharmaceutically acceptable carrier.

[0203] In certain embodiments, the pharmaceutical composition of the present invention further comprises at least one additional pharmaceutically active agent other than the compound of the present invention.

[0204] The pharmaceutical composition of the present invention can be prepared by combining one or more compounds of the present invention with a pharmaceutically acceptable carrier and optionally one or more additional pharmaceutically active agents.

[0205] As stated above, an "effective amount" means any amount sufficient to achieve the desired biological effect. In light of the teachings provided herein, a prophylactic or therapeutic treatment regimen that does not cause substantial unwanted toxicity but is effective in treating a particular subject can be devised by choosing among the various active compounds and weighing factors such as potency, relative bioavailability, patient body weight, severity of adverse side effects, and mode of administration. The effective amount for any particular administration can vary depending on factors such as the disease or disorder being treated, the particular compound of the present invention being administered, the size of the subject, or the severity of the disease or disorder. A person of ordinary skill in the art can determine the effective amount of a particular compound of the present invention and / or other therapeutic agents empirically, without undue experimentation. The maximum dose, i.e., the highest safe dose according to some medical diagnoses, can be used. Multiple daily doses can be considered to achieve an appropriate systemic level of the compound. The appropriate systemic level can be determined, for example, by measuring the peak or sustained plasma levels of the drug in the patient. "Dose / dosage" is used interchangeably herein.

[0206] In certain embodiments, the intravenous administration of the compound can generally be from 0.1 mg / kg / day to 20 mg / kg / day. In one embodiment, the intravenous administration of the compound can generally be from 0.1 mg / kg / day to 2 mg / kg / day. In one embodiment, the intravenous administration of the compound can generally be from 0.5 mg / kg / day to 5 mg / kg / day. In one embodiment, the intravenous administration of the compound can generally be from 1 mg / kg / day to 20 mg / kg / day. In one embodiment, the intravenous administration of the compound can generally be from 1 mg / kg / day to 10 mg / kg / day.

[0207] Typically, for human subjects, the daily oral dose of the compound will be from about 0.01 mg / kg / day to 1000 mg / kg / day. Oral doses in the range of 0.5 to 50 mg / kg administered once or more times per day are expected to produce therapeutic results. The dose may be adjusted appropriately to achieve the desired local or systemic drug level, depending on the mode of administration. For example, it is expected that the daily dose for intravenous administration will be reduced by one to several orders of magnitude. If the response of the subject at such a dose is insufficient, higher doses (or effectively higher doses by different, more localized delivery routes) may be used within the limits of patient tolerance. Multiple daily doses may be considered to achieve the appropriate systemic level of the compound.

[0208] For any of the compounds described herein, a therapeutically effective amount can be initially determined in animal models. The therapeutically effective dose can also be determined based on human data for compounds that have been tested in humans and compounds known to exhibit similar pharmacological activity (such as other related active agents). Parenteral administration may require higher doses. The dose administered can be adjusted based on the relative bioavailability and potency of the compound administered. Adjusting the dose to achieve maximum efficacy based on the methods described above and other methods known in the art is well within the capabilities of a person of ordinary skill in the art.

[0209] The formulations of the present invention can be administered in the form of a pharmaceutically acceptable solution which may routinely contain pharmaceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants and optionally other therapeutic ingredients.

[0210] For use in therapy, an effective amount of the compound can be administered to a subject by any mode of delivering the compound to the desired surface. Administration of the pharmaceutical composition can be accomplished by any means known to those skilled in the art. Routes of administration include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (urinary bladder), oral, subcutaneous, direct injection (e.g., injection into a tumor or abscess), mucosal (e.g., topical to the eye), inhalation, and topical.

[0211] For intravenous and other parenteral routes of administration, the compounds of the present invention can be formulated as lyophilized preparations, lyophilized preparations of liposome-embedded or encapsulated active compounds, lipid complexes in aqueous suspensions, or salt complexes. Lyophilized formulations are generally reconstituted in a suitable aqueous solution, such as sterile water or saline, shortly before administration.

[0212] For oral administration, the compounds can be readily formulated by combining one or more active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by a subject to be treated. Oral pharmaceutical preparations can be obtained as solid excipients, optionally grinding the resulting mixture, and, if desired, processing the granule mixture with suitable auxiliaries to obtain tablets or dragee cores. Suitable excipients are in particular fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate. Optionally, the oral formulations can also be formulated in saline or buffer (e.g., EDTA for neutralizing internal acidic conditions), or can be administered without any carrier.

[0213] Also specifically contemplated are oral dosage forms of one or more of the above components. The one or more components can be chemically modified to render the oral delivery of the derivatives effective. Generally, the chemical modifications contemplated are the attachment of at least one moiety to the component molecule itself, wherein the moiety permits (a) inhibition of acid hydrolysis; (b) absorption from the stomach or intestine into the bloodstream. It is also desirable to increase the overall stability of one or more components and increase the in vivo circulation time. 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, N.Y., 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-trioxocane. For the pharmaceutical uses indicated above, polyethylene glycol moieties are suitable.

[0214] For a component (or derivative), the release location can be the stomach, small intestine (duodenum, jejunum or ileum), or large intestine. Those skilled in the art can obtain formulations that do not dissolve in the stomach but release substances elsewhere in the duodenum or intestine. Preferably, by protecting the compound (or derivative) of the present invention or by releasing the bioactive substance outside the gastric environment, such as in the intestine, the release will avoid the harmful effects of the gastric environment.

[0215] To ensure complete gastric juice tolerance, a coating that is at least impermeable at pH 5.0 is essential. Examples of more common inert ingredients used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can be used as mixed films.

[0216] Coatings or coating mixtures can also be used on tablets that are not intended to protect the stomach. The coating can include a sugar coating or a coating that makes the tablet easier to swallow. Capsules may consist of a hard shell (such as gelatin) for delivering dry therapeutic agents (such as powders); for liquid forms, soft gelatin shells can be used. The outer shell material of cachets can be thick starch or other edible paper. For pills, lozenges, molded tablets or die-stamped tablets, moist massing techniques can be used.

[0217] The therapeutic agent can be included in the formulation as fine multi-microparticles in the form of granules or pellets with a particle size of about 1 mm. The formulation of materials for capsule administration can also be powders, slightly compressed plugs or even tablets. The therapeutic agent can be prepared by pressing.

[0218] Both coloring agents and flavoring agents can be included. For example, the compound (or derivative) of the present invention can be formulated (such as by encapsulation in liposomes or microspheres) and then further included in an edible product, such as a refrigerated beverage containing coloring agents and flavoring agents.

[0219] The therapeutic agent can be diluted with inert materials or the volume of the therapeutic agent can be increased. These diluents may include carbohydrates, especially mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextran and starch. Certain inorganic salts can also be used as fillers including calcium phosphate tribasic, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell.

[0220] Disintegrants can be included in the formulation of therapeutic agents to form solid dosage forms. Materials used as disintegrants include, but are not limited to, starches, including the starch-based commercial disintegrant Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethyl cellulose, hyperbranched starch, sodium alginate, gelatin, orange peel, acidic carboxymethyl cellulose, natural sponge, and bentonite can all be used. Another form of disintegrant is insoluble cation exchange resin. Powdered gums can be used as disintegrants and binders, and these disintegrants and binders can include powdered gums such as agar, karaya gum, or tragacanth gum. Alginic acid and its sodium salt can also be used as disintegrants.

[0221] Binders can be used to hold the therapeutic agent together to form hard tablets and include materials from natural products such as gum arabic, tragacanth gum, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can both be used in alcohol solutions to granulate the therapeutic agent.

[0222] Antifriction agents can be included in the formulation of therapeutic agents to prevent adhesion during the formulation process. Lubricants can be used as a layer between the therapeutic agent wall and the die wall, and these lubricants can include, but are not limited to: stearic acid, including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oil, and wax. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, Carbowax 4000, and 6000 can also be used.

[0223] Glidants may be added, which may modify the flow properties of the drug during the formulation period and assist in rearrangement during compression. Glidants can include starch, talc, pyrogenic silica, and hydrated silicoaluminate.

[0224] To assist the therapeutic agent in dissolving into an aqueous environment, surfactants can be added as wetting agents. Surfactants can include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate, and sodium dioctyl sulfonate. Available cationic detergents can include benzalkonium chloride and benzethonium chloride. Potential nonionic detergents that can be included as surfactants in the formulation include polidocanol 400, polyethylene glycol 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid ester, methylcellulose, and carboxymethylcellulose. These surfactants can be present in the formulation of the compounds or derivatives of the present invention alone or as mixtures in different proportions.

[0225] Oral pharmaceutical preparations include push-fit capsules made of gelatin and soft-sealed capsules made of gelatin and a plasticizer (such as glycerol or sorbitol). The push-fit capsules may contain an active ingredient mixed with a filler (such as lactose), a binder (such as starch), and / or a lubricant (such as talc or magnesium stearate), and optionally a stabilizer. In the soft capsules, the active compound may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin, or liquid polyethylene glycol. Additionally, a stabilizer may be added. Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All formulations for oral administration should be in a dosage suitable for such administration.

[0226] For buccal administration, the composition may take the form of tablets or lozenges formulated in a conventional manner.

[0227] For topical administration, the compound may be formulated as a solution, gel, ointment, cream, suspension, etc., as is well known in the art. Systemic formulations include those designed for administration by injection (e.g., subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection), and those designed for transdermal, transmucosal oral, or pulmonary administration.

[0228] For administration by inhalation, the compounds used according to the present invention may be conveniently delivered in the form of an aerosol spray by a pressurized pack or a nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gases). In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve for delivering a metered amount. Capsules and cartridges, such as made of gelatin, containing a powder mixture of the compound with a suitable powder matrix (such as lactose or starch) for use in an inhaler or insufflator may be formulated.

[0229] The present invention also contemplates pulmonary delivery of the compounds (or salts thereof) disclosed 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 II, Keystone, Colorado, March, (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon γ and tumor necrosis factor α) 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 to achieve a systemic effect are described in U.S. Patent No. 5,451,569, issued September 19, 1995 to Wong et al. (incorporated by reference).

[0230] Contemplated for use in practicing the present invention are a wide range of mechanical devices (including but not limited to nebulizers, metered dose inhalers, and powder inhalers) designed for pulmonary delivery of therapeutic products, all of which are familiar to those of skill in the art.

[0231] Some specific examples of commercially available devices suitable for practicing the present invention are the Ultravent nebulizer, manufactured by Mallinckrodt, Inc., St. Louis, Mo.; the Acorn II nebulizer, manufactured by Marquest Medical Products, Englewood, Colo.; the Ventolin metered dose inhaler, manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and the Spinhaler powder inhaler, manufactured by Fisons Corp., Bedford, Mass.

[0232] All such devices require the use of formulations suitable for dispensing the compounds of the present invention. Generally, each formulation is specific to the type of device used and may involve the use of suitable propellant materials in addition to the common diluents, adjuvants, and / or carriers used in therapy. In addition, the use of liposomes, microcapsules or microspheres, inclusion complexes or other types of carriers is contemplated. The chemically modified compounds of the present invention can also be prepared in different formulation forms depending on the type of chemical modification or the type of device used.

[0233] Formulations suitable for use with jet or ultrasonic nebulizers generally contain the compound (or derivative) of the present invention dissolved in water at a concentration of about 0.1 mg to 25 mg of the bioactive compound of the present invention per milliliter of solution. The formulation may also include buffers and monosaccharides (e.g., for the stabilization and osmotic pressure regulation of the inhibitor). Nebulizer formulations may also contain surfactants to reduce or prevent surface-induced aggregation of the compounds of the present invention caused by solution atomization during aerosol formation.

[0234] Formulations for use with metered-dose inhaler devices generally contain fine powders 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 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 soy lecithin. Oleic acid can also be used as a surfactant.

[0235] Formulations for dispensing from powder inhaler devices will contain fine dry powders containing the compound (or derivative) of the present invention and may also include amounts (e.g., 50 wt% to 90 wt% of the formulation) of bulking agents, such as lactose, sorbitol, sucrose or mannitol, that facilitate the dispersion of the powder from the device. The compound (or derivative) of the present invention should advantageously be prepared in the form of microparticles having an average particle size of less than 10 micrometers (μm), most preferably 0.5 to 5 μm, for most effective delivery to the deep lung.

[0236] Nasal delivery of the pharmaceutical compositions of the present invention is also contemplated. Nasal delivery allows the pharmaceutical compositions of the present invention to enter the bloodstream directly after nasal administration of the therapeutic product without the product depositing in the lungs. Formulations for nasal delivery include those having dextran or cyclodextrin.

[0237] For nasal administration, a useful device is a small, rigid bottle to which a metered sprayer is attached. In one embodiment, a metered dose is delivered by inhaling the pharmaceutical composition solution of the present invention into a chamber of defined volume having apertures sized to atomize and aerosolize the formulation by forming a spray when the liquid in the chamber is compressed. The chamber is compressed to administer the pharmaceutical composition of the present invention. In a particular embodiment, the chamber is of a piston arrangement. Such devices are commercially available.

[0238] Alternatively, a plastic squeeze bottle with apertures or openings is used, sized to atomize the aerosol formulation by forming a spray when squeezed. The opening is typically located at the top of the bottle, and the top is usually tapered to fit partially into the nasal passage to effectively administer the aerosol formulation. Preferably, the nasal inhaler will provide a metered amount of the aerosol formulation for administering the measured dose of the drug.

[0239] When systemic delivery of the compound is desired, the compound can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Injectable formulations can be presented in unit dosage forms, e.g., in ampoules or multi-dose containers with added preservatives. The composition can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulating agents such as suspending, stabilizing and / or dispersing powders.

[0240] Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compound in water-soluble form. Additionally, suspensions of the active compound can be prepared as appropriate 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, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound to allow the preparation of highly concentrated solutions.

[0241] Alternatively, the active compound can be in the form of a powder to be reconstituted with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0242] The compound can also be formulated as rectal or vaginal compositions, such as, for example, suppositories or retention enemas containing conventional suppository bases such as cocoa butter or other glycerides.

[0243] In addition to the formulations described above, the compound can also be formulated as a depot formulation. Such long-acting formulations can be formulated with suitable polymeric or hydrophobic materials (e.g., formulated as an emulsion in an acceptable oil) or ion exchange resins, or formulated as a slightly soluble derivative, e.g., as a slightly soluble salt.

[0244] The pharmaceutical composition may also comprise a suitable solid or gel phase carrier or excipient. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

[0245] Suitable liquid or solid pharmaceutical dosage forms are, for example, aqueous or saline solutions for inhalation, which are microencapsulated, embedded, coated onto fine gold particles, contained in liposomes, atomized, aerosols for implantation into the skin, pellets, or dried onto sharp objects for scratching into the skin. The pharmaceutical composition also includes granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or formulations of slow-release active compounds, and excipients and additives and / or adjuvants such as disintegrants, binders, coating agents, swelling agents, lubricants, flavorants, sweeteners, or solubilizers are typically used as described above in the preparation of the pharmaceutical composition. The pharmaceutical composition is suitable for a variety of drug delivery systems. For a brief review of methods for drug delivery, see Langer R, Science 249: 1527-33 (1990).

[0246] The compounds of the invention and optionally other therapeutic agents can be administered in themselves (pure) or in the form of pharmaceutically acceptable salts or co-crystals. When used in medicine, the salt or co-crystal should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts or co-crystals can be conveniently used to prepare their pharmaceutically acceptable salts or co-crystals. Such salts include, but are not limited to, salts 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. In addition, such salts can be prepared as alkali metal or alkaline earth metal salts, such as sodium, potassium, or calcium salts of carboxylic acid groups.

[0247] 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).

[0248] The pharmaceutical composition of the present invention contains an effective amount of a compound as described herein and optionally a therapeutic agent contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers, diluents or encapsulating substances which are suitable for administration to humans or other vertebrates. The term "carrier" denotes a natural or synthetic organic or inorganic component which is combined with the active ingredient to facilitate its application. The components of the pharmaceutical composition are also capable of being admixed with the compounds of the present invention and with each other in such a way that there is no interaction which would substantially impair the desired pharmaceutical efficacy.

[0249] One or more therapeutic agents, specifically including but not limited to the compounds of the present invention, may be provided in particulate form. As used herein, particulate means nanoparticles or microparticles (or in some cases larger particles) which may consist in whole or in part of the compounds of the present invention or one or more other therapeutic agents as described herein. The particles may contain one or more therapeutic agents located in a core surrounded by a coating (including but not limited to enteric coating). The one or more therapeutic agents may also be dispersed throughout the particles. The one or more therapeutic agents may also be adsorbed into the particles. The particles may have any level of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release and any combination thereof, etc. In addition to the one or more therapeutic agents, the particles may include any of those materials conventionally used in the pharmaceutical and medical arts (including but not limited to erodible, non-erodible, biodegradable or non-biodegradable materials or combinations thereof). The particles may be microcapsules containing the compounds of the present invention in solution or semi-solid state. The particles may actually be of any shape.

[0250] Both non-biodegradable and biodegradable polymeric materials can be used to manufacture particles for delivering the one or more therapeutic agents. Such polymers can be natural polymers or synthetic polymers. The polymers are selected based on the desired period of release. Particularly interesting bioadhesive polymers include the bioerodible hydrogels described in Sawhney H S et al. (1993) Macromolecules 26: 581-7, the teachings of which are incorporated herein. These polymers include polyhyaluronic acid, casein, gelatin, gelatin protein, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate) and poly(octadecyl acrylate).

[0251] The one or more therapeutic agents may be included in a controlled release system. The term "controlled release" means any medicated formulation in which the manner and profile of release of the drug from the formulation are controlled. This refers to immediate release and non-immediate release formulations, where non-immediate release formulations include, but are not limited to, sustained release and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in its conventional sense to mean a medicated formulation that provides for the gradual release of a drug over an extended period of time and preferably, but not necessarily, results in a substantially constant blood level of the drug over the extended period of time. The term "delayed release" is used in its conventional sense to mean a medicated formulation in which there is a time delay between the administration of the formulation and the release of the drug therefrom. "Delayed release" may or may not involve the gradual release of the drug over an extended period of time and thus may or may not be "sustained release".

[0252] The use of long-term sustained release implants may be particularly suitable for the treatment of chronic conditions. As used herein, "long-term" release means that the implant is constructed and arranged to deliver a therapeutic level of the active ingredient for at least 7 days and preferably 30 - 60 days. Long-term sustained release implants are well known to those of ordinary skill in the art and include some of the release systems described above.

[0253] Those of ordinary skill in the relevant art will appreciate that other suitable modifications and adaptations of the compositions and methods described herein will be apparent from the description of the invention contained herein in view of the information known to those of ordinary skill in the art, and can be made without departing from the scope of the invention or any of its embodiments. The invention has now been described in detail, and the invention will be more clearly understood by reference to the following examples, which are included herein for illustrative purposes only and are not intended to limit the invention.

[0254] Examples

[0255] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0256] Example 1: SLC6A19 Isoleucine Transport Assay

[0257] Generation and maintenance of cell lines

[0258] Flp-In TM T-REx TMThe 293 cell line was purchased from Thermo Fisher Scientific. This cell line was used to generate a stable cell line that inducibly expresses human SLC6A19 with a C-terminal V5 tag and stably expresses human TMEM27 (also known as Collectrin) with a C-terminal myc-DDK tag. This stable cell line was generated by transfecting plasmids encoding SLC6A19 and TMEM27 using standard protocols followed by antibiotic selection. The stable cells were maintained in DMEM / F12 supplemented with Glutamax, 10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin, 200 μg / mL hygromycin, 10 μg / mL blasticidin, and 300 μg / mL neomycin (Thermo Fisher).

[0259] Assays: Isoleucine transport assay in 96-well plate format

[0260] On day 0, the stable cell line was seeded at a density of 35,000 cells per well in a poly-D-lysine-coated 96-well cell culture-treated plate. On day 1, the expression of SLC6A19 was induced by dispensing tetracycline at a final concentration of 1 μg / mL using a Tecan D300e digital dispenser. The transport assay was run on day 2. The medium was removed from the plate using the GentleSpin setting of a Centrifugal BlueWasher (Blue Cat Bio), and then the cells were washed with 175 μL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, the cells were incubated with 70 μL of Krebs buffer (140 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl 2、1.2 mMMgCl 2、11Cells were treated with DMSO, positive control, or compound diluted in 10 mM HEPES, 10 mM glucose, pH 7.4. After 20 - 60 minutes, 30 μL of 3.3 mM 13C6,15N-L-isoleucine solution (Cambridge Isotope Laboratories) was added. After incubating with the isoleucine substrate for 20 min at room temperature, the cells were washed with 175 μL of live cell imaging solution using a Blue Washer. The cells were then lysed in 150 μL of 15 μM D-leucine-d10 (CDN Isotopes) in ultrapure water. The plate was placed on an oscillator at 700 rpm for at least 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 oscillator for at least 2 minutes to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000 rpm for 5 min to pellet cell debris and precipitate. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0261] Assays: Isoleucine transport assay in 384-well plate format

[0262] On day 0, a stable cell line was seeded at a density of 20,000 cells per well in a medium containing 1 μg / mL tetracycline in a poly-D-lysine-coated, cell culture-treated 384-well plate using a Viaflo 384-well pipettor. The transport assay was run the following day (day 1). The medium was removed from the plate using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio), and the cells were then washed with 80 μL of live cell imaging solution (ThermoFisher) using a Blue Washer. After washing, 20 μL of Krebs buffer (140 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl 2、1.2 mMMgCl 2、11Cells were treated with DMSO, positive control, or compound diluted in 10 mM HEPES, 10 mM glucose, pH 7.4). After incubation at room temperature for 20 - 60 minutes, 8.6 uL of 3.3 mM 13C6,15N-L-isoleucine solution (Cambridge Isotope Laboratories) was added. After incubation with the isoleucine substrate at room temperature for 20 min, the cells were washed with 80 uL of live cell imaging solution using a Blue Washer. The cells were then lysed in 80 uL of 15 uM D-leucine-d10 (CDN Isotopes) in ultrapure water. The plate was placed on an orbital shaker at 700 rpm for at least 2 hours 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 at least 5 minutes to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000 rpm for 10 min to pellet cell debris and precipitate. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0263] 13C6,15N-L-isoleucine analysis was performed using a RapidFire365-QTOF 6545 (Agilent). Quantitative sample analysis utilized automated solid-phase extraction (HILIC H6 cartridge) prior to mass spectrometry injection. Samples were loaded using 95% acetonitrile, 0.1% formic acid, and eluted directly from the cartridge using 5% acetonitrile, 0.1% formic acid for ESI-MS (electrospray ionization) analysis. Analytes were quantified using Agilent Masshunter Quant software based on high-resolution full-scan data.

[0264] Example 2: Synthesis of Exemplary Compounds

[0265] Procedure 1: Synthesis of (S)-1-(tert-butoxycarbonyl)-3-(trifluoromethyl)pyrrolidine-3-carboxylic acid

[0266]

[0267] Step 1: At 0 °C, TFA (1.22 g, 10.7 mmol) was added dropwise to a mixture of 2-(trifluoromethyl)acrylic acid (15.0 g, 107.1 mmol) and N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methanamine (25.4 g, 107.1 mmol) in DCM (150 mL). The resulting mixture was stirred at room temperature for 17 hours. The mixture was then diluted with petroleum ether (600 mL) and filtered after stirring at room temperature for 30 minutes. The filter cake was then washed with a solution of PE:EtOAc = 3:1 (60 mL) and concentrated in vacuo to give A1 (24.5 g) as a white solid.

[0268] The enantiomers of A1 can be separated by chiral SFC (Shimadzu E-UC SFC; CHIRALPAK IC, 5 * 25 cm, 5 μm) to obtain (R)-A1 (10.2 g, 34.85% yield) and (S)-A1 (12.1 g, 41.35% yield) as white solids. LC / MS (ESI) m / z: 274 (M + H) + 。

[0269] Step 2: Add 20% Pd / C (1.6 g, weight / weight) to a solution of (S)-A1 (8.0 g, 29.28 mmol) in MeOH (150 mL). Stir the resulting mixture at room temperature under an H 2 atmosphere for 2 hours. Then filter the mixture and concentrate the filtrate under vacuum to obtain (S)-3-(trifluoromethyl)pyrrolidine-3-carboxylic acid (5.1 g, 95.12% yield) as a pale yellow oil, which is used directly in the next step without further purification. LC / MS (ESI) m / z: 184 (M + H) + 。

[0270] Step 3: At 0 °C, add Boc 2 O (6.69 g, 30.63 mmol) dropwise to a mixture of (S)-3-(trifluoromethyl)pyrrolidine-3-carboxylic acid (5.1 g, 27.85 mmol) and TEA (5.64 g, 55.70 mmol) in DCM (70 mL). Stir the resulting mixture at room temperature for 2 hours. Then concentrate the mixture under vacuum and purify the crude product by silica gel column chromatography (DCM:MeOH = 100:0 to 12:1) to obtain 5 (7.5 g, 95.08% yield) as a colorless oil. LC / MS (ESI) m / z: 282 (M - H)-.

[0271] Procedure 2: Synthesis of arylamidoximes

[0272] The arylamidoximes mentioned below can be prepared from appropriately substituted aryl nitriles according to appropriate modifications of the following procedure.

[0273] Synthesis of 4-cyano-N-hydroxybenzamide (B1)

[0274]

[0275] To a mixture of terephthalonitrile (12.8 g, 99.90 mmol) and TEA (11.12 g, 109.88 mmol) in EtOH (250 mL) was added hydroxylamine hydrochloride (6.94 g, 99.90 mmol). The resulting mixture was stirred at 70 °C for 2 h. After consumption of the SM, the mixture was concentrated in vacuo and the crude product was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 10:1) to give B1 as a yellow solid (11.20 g, 69.57% yield). LC / MS (ESI) m / z: 162 (M+H) + 。

[0276] Procedure 3: (S)-3-Aryl-5- ( 3- ( Synthesis of (trifluoromethyl)pyrrolidin-3-yl)-1,2,4-oxadiazole

[0277] The C2 type 1,2,4-oxadiazoles mentioned below can be prepared from substituted arylamidoximes such as B1 and (S)-A2 or racemic A2 according to appropriate modifications of the following procedure.

[0278]

[0279] Synthesis of 4-cyano-N-hydroxybenzamide (C 2 )

[0280] Step 1: To a solution of (S)-A2 (7.5 g, 26.48 mmol) in DMF (250 mL) was added CDI (6.44 g, 39.72 mmol). The mixture was stirred at 85 °C for 1 h. Then B1 (5.50 g, 34.42 mmol) was added to the above mixture. The resulting mixture was stirred at 85 °C for an additional 10 h. After cooling, the mixture was diluted with water (100 mL) and washed twice with EtOAc (60 mL). The combined organic layers were separated, washed with saturated NH 4 Cl solution (80 mL) and brine (80 mL), dried over anhydrous Na 2 SO 4 and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:0 to 8:1) to give C1 as a colorless oil (9.60 g, 88.78% yield). LC / MS (ESI) m / z: 409 (M+H) + 。

[0281] Step 2: At 0 °C under N 2Under an atmosphere, C1 (9.60 g, 23.51 mmol) was added portionwise to a 4N HCl / dioxane solution (120 mL). The resulting mixture was stirred at room temperature for 2 hours. Then the mixture was concentrated under reduced pressure to obtain crude C2 (7.25 mg, 89.51% yield) as a pale yellow oil, which was used directly in the next step without further purification. LC / MS (ESI) m / z: 309 (M+H) + .

[0282] Examples 1 - 42: The compounds in the following table can be prepared from substituted pyrrolidines such as C2 and appropriate Boc-protected diamines such as D1 according to suitable modifications of the following procedure.

[0283]

[0284] Step 1: At 0 °C, CDI (4.58 g, 28.22 mmol) was added to a mixture of C2 (5.65 g, 28.22 mmol) and DIEA (9.12 g, 70.56 mmol) in DMF (80 mL). The mixture was stirred at room temperature for 30 min. Then D1 (7.25 g, 23.51 mmol) was added to the above mixture. The resulting mixture was stirred at 50 °C for 3 hours. After cooling, the mixture was quenched with an aqueous HCl (120 mL, 2N) solution and extracted twice with EtOAc (100 mL). The combined organic layers were separated, washed with brine (120 mL), dried over anhydrous Na 2 SO 4 and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 18:1) to obtain D2 (11.6 g, 92.27% yield) as an off-white solid. LC / MS (ESI) m / z: 435 (M - 100 + H) + .

[0285] Step 2: At 0 °C under an N 2 atmosphere, D2 (11.6 g, 21.7 mmol) was added to a solution of 4N HCl / dioxane (120 mL). The resulting mixture was stirred at room temperature for 2 hours. Then the mixture was concentrated under reduced pressure to obtain a crude product (10.6 g) as a pale yellow oil. The crude product was then diluted with MTBE (100 mL) and the resulting slurry was filtered after stirring at room temperature for 5 hours. The filter cake was washed with MTBE (100 mL) and concentrated in vacuo to obtain D3 (7.10 g, 75.31% yield) as a white solid.

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301] Examples 43 - 66: The compounds in the following table can be prepared from a substituted pyrrolidine (such as E2, which is itself prepared according to Procedure 3) and a suitable diamine such as E1 according to a suitable modification of the following procedure.

[0302]

[0303] Step 1: At 0 °C, CDI (82 mg, 0.572 mmol) was added portionwise to a mixture of E2 (156 mg, 0.440 mmol) and DIEA (74 mg, 0.572 mmol) in DMF (1 mL). The reaction mixture was stirred at room temperature for 40 min. Then E1 (88 mg, 0.88 mmol) was added to the above mixture, and the resulting mixture was stirred at 50 °C for 3 h. Then the mixture was concentrated to dryness. The residue was purified by preparative reverse-phase HPLC to give E3 as a colorless oil. LC / MS (ESI) m / z: 444 (M + H) + 。

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314] Examples 67 - 73: The compounds in the following table can be prepared from substituted pyrrolidines such as C2 and appropriate diamines such as F1 according to suitable modifications of the following procedure.

[0315]

[0316] Step 1: CDI (33 mg, 0.234 mmol) was added to a mixture of C2 (40 mg, 0.19 mmol) and DIEA (34 mg, 0.259 mmol) in DMF (0.5 mL). The reaction mixture was stirred at room temperature for 15 min. Then F1 (40 mg, 0.194 mmol) was added to the above mixture, and the resulting mixture was stirred at 50 °C for 3 h. Then the mixture was concentrated to dryness. The residue was taken up in 4N HCl / dioxane solution (1 m) and stirred for 30 min, then concentrated to dryness again to give F2, which was used without further purification. LC / MS (ESI) m / z: 439 (M + H) + 。

[0317] Step 2: DIPEA (38 mg, 292 μmol) and 37% formaldehyde solution (22 μL, 292 μmol) were added to a solution of F2 HCl (43 mg, 98 μmol) in methanol (1.5 mL). Sodium triacetoxyborohydride (42 mg, 195 μmol) was added and the solution was stirred at room temperature. Additional portions of sodium triacetoxyborohydride were added until LCMS indicated complete consumption of the starting material. The resulting solution was purified directly by preparative reverse-phase HPLC to give F3 as a colorless oil. LC / MS (ESI) m / z: 453 (M + H) + 。

[0318]

[0319]

[0320]

[0321] Examples 74 - 76: The compounds in the following table can be prepared from 1-(tert-butoxycarbonyl)-4-(trifluoromethyl)piperidine-4-carboxylic acid, an appropriate diamine, and an appropriate aryl nitrile by a method similar to the preparation of the above pyrrolidine.

[0322]

[0323]

[0324] Procedure 3: Synthesis of (S)-3-aryl-5-(3-(trifluoromethyl)pyrrolidin-3-yl)-1,2,4-oxadiazole

[0325] The G5 type 1,2,4-oxadiazole mentioned below can be prepared from nitrile G1 and an appropriately substituted benzoate such as G3 according to a suitable modification of the following procedure.

[0326]

[0327] Step 1: Add hydroxylamine hydrochloride (585 mg, 8.42 mmol) and triethylamine (1.17 mL, 8.42 mmol) to a solution of G1 (1.07 g, 4.21 mmol) in ethanol (10 mL). Heat the solution at 60 °C for two hours and then cool to room temperature. Dilute the solution with water and ethyl acetate and separate the phases. Extract the aqueous phase with ethyl acetate more than twice, then wash the combined organic phases with water, saturated NaCl aqueous solution, dry over MgSO4 and concentrate to obtain G2 (1.3 g, 4.53 mmol) as a white solid, which can be used without further purification. LC / MS (ESI) m / z: 288.0 (M + H) + .

[0328] Step 2: At room temperature, add DCC (256 mg, 1.24 mmol) to a mixture of G2 (324 mg, 1.13 mmol) and G3 (176 mg, 1.13 mmol) in dioxane (3 mL). Then heat the solution at 90 °C overnight and then cool to room temperature. Remove the precipitate by filtration, and concentrate the filtrate to an oil, which is then purified by column chromatography (0 - 100% ethyl acetate / heptane) to obtain the desired oxadiazole G4 (125.4 mg) as a colorless residue. LC / MS (ESI) m / z: 399.1 (M + H) + .

[0329] Step 3: At room temperature, add chloroethyl chloroformate (67 μL, 0.614 mmol) to a solution of G4 (125.4 mg, 0.294 mmol) in DCE (1 mL). Stir the solution at 50 °C for 75 minutes, then add methanol (1 mL), and heat the solution at 60 °C for one hour. Then concentrate the solution to obtain G5HCl, which can be used without further purification. LC / MS (ESI) m / z: 309.1 (M+H) + 。

[0330] Examples 77 - 79: The compounds in the following table can be prepared from G5 type 1,2,4-oxadiazole and appropriate diamines by suitable modification of the above method.

[0331]

[0332]

[0333] Example 80: Example 80 was prepared according to the following method.

[0334]

[0335] Step 1: Add DIPEA (199 mg, 1.54 mmol) and 2-bromo-1-(4-bromophenyl)ethan-1-one (213 mg, 0.77 mmol) to a mixture of A1 (210 mg, 0.77 mmol) in DMF (5 mL). Stir the resulting mixture at room temperature for 3 hours. Then dilute the mixture with water (20 mL) and extract twice with EtOAc (15 mL). Wash the combined organic layers with saturated NH 4 Cl solution and brine, dry over anhydrous Na 2 SO 4 and concentrate to dryness. Purify the residue by silica gel column chromatography (eluting with PE:EtOAc = 100:0 to 40:1) to obtain H1 (350 mg, 97.01% yield) as a colorless oil. LC / MS (ESI) m / z: 470 / 472 (M+H) + 。

[0336] Step 2: Add acetamide (1.28 g, 21.75 mmol) and boron trifluoride diethyl etherate (1.5 mL, 0.73 mmol) to a solution of H1 (350 mg, 0.75 mmol) in toluene (15 mL). Stir the resulting mixture in a sealed tube at 150 °C for 6 hours. Continue the reaction until TLC indicates complete consumption of the starting material (PE:EtOAc = 10:1). Then cool the mixture to 0 °C and wash with saturated NaHCO 3The solution (40 mL) was quenched and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine, dried over anhydrous Na 2 SO 4 and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:0 to 100:3) to give H2 as a colorless oil (135 mg, 40.20% yield). LC / MS (ESI) m / z: 451 / 453 (M+H) + .

[0337] Step 3: To a mixture of H2 (135 mg, 0.30 mmol) and Zn(CN) 2 (53 mg, 0.45 mmol) in DMF (7 mL) was added Pd(PPh 3 ) 4 (35 mg, 0.03 mmol), and the resulting mixture was stirred at 120 °C under N 2 atmosphere for 16 h. Then the mixture was diluted with H 2 O (20 mL) and extracted twice with EtOAc (15 mL). The combined organic layers were washed with saturated NH 4 Cl solution and brine, dried over anhydrous Na 2 SO 4 and concentrated to dryness. The residue was purified by silica gel column chromatography (eluting with PE:EtOAc = 100:1 to 4:1) to give H3 as a colorless oil (83 mg, 69.82% yield). LC / MS (ESI) m / z: 398 (M+H) + .

[0338] Step 4: To a solution of H3 (83 mg, 0.21 mmol) in DCM (5 mL) was added chloroethyl chloroformate (90 mg, 0.63 mmol), and the resulting mixture was stirred at 50 °C under 2 atmosphere for 20 h. Then MeOH (3 mL) was added to the above mixture and the mixture was stirred at 75 °C for another 2 h. Then the mixture was concentrated to give crude H4HCl as a colorless oil (64 mg, 99.71% yield). LC / MS (ESI) m / z: 308 (M+H) + .

[0339] Step 5: To a solution of D1HCl (55 mg, 0.27 mmol) in DMF (4 mL) was added DIEA (76 mg, 0.59 mmol) and CDI (44 mg, 0.27 mmol), and the mixture was stirred at 0 °C for 30 min. Then H4HCl (64 mg, 0.21 mmol) was added to the above mixture, and the resulting mixture was stirred at 50 °C for another 2 h. Then the mixture was diluted with H 2 O (20 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were washed with saturated NH 4 Cl solution and brine, dried over anhydrous Na 2 SO 4 and concentrated to dryness. The residue was purified by silica gel column chromatography (DCM:MeOH = 100:0 to 100:3) to afford Boc-F5 as a colorless oil (70 mg, 63.02% yield). LC / MS (ESI) m / z: 534 (M+H) + . At 0 °C, Boc-H5 (70 mg, 0.13 mmol) and HCl / dioxane (4 mol / L, 4 mL) were charged into a round-bottom flask. The resulting mixture was stirred at room temperature for 1 h. LCMS indicated complete consumption of the starting material. Then the mixture was concentrated to dryness under reduced pressure. The residue was purified via preparative HPLC to afford H5 as a white solid (39.0 mg, 68.55% yield).

[0340]

[0341] Example 81: Example 81 was prepared according to the following method.

[0342]

[0343] Step 1: At 0 °C, to a mixture of (rac)-A2 (5.18 g, 18.29 mmol) and DIEA (7.09 g, 54.86 mmol) in MeCN (100 mL) was added CDI (4.45 g, 27.43 mmol). The mixture was stirred at 90 °C for 1 h. Then N,O-dimethylhydroxylamine hydrochloride (2.32 g, 23.77 mmol) was added to the above mixture. The resulting mixture was stirred at 90 °C for another 3 h. After cooling, the mixture was quenched with aqueous HCl solution (100 mL, 2 N) and extracted twice with dichloromethane (100 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous Na 2 SO 4Dry and concentrate under reduced pressure to dryness. Purify the residue by silica gel column chromatography (eluting with PE:EtOAc = 100:0 to 4:1) to obtain I1 as a white solid (5.09 g, 85.29% yield). LC / MS (ESI) m / z: 271 (M - 56 + H) + .

[0344] Step 2: At -78 °C under N 2 atmosphere, add dropwise a solution of DIBAL-H in THF (18.4 mL, 1 M) to a solution of I1 (2.0 g, 6.13 mmol) in anhydrous DCM (50 mL). Stir the resulting mixture at -78 °C for 1 h. Then quench the mixture portionwise with a saturated solution of sodium potassium tartrate tetrahydrate (50 mL) at 0 °C and stir at room temperature for 1 h. Then extract the mixture twice with DCM (60 mL). Wash the combined organic layers with brine (100 mL), dry over anhydrous Na 2 SO 4 dry and concentrate under reduced pressure to dryness. Purify the crude product by silica gel column chromatography (eluting with PE:EtOAc = 100:0 to 20:1) to obtain I2 as a white solid (1.50 g, 91.58% yield). LC / MS (ESI) m / z: 212 (M - 56 + H) + .

[0345] Step 3: Add K 2 CO 3 (2.33 g, 16.84 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (1.62 g, 8.42 mmol) to a solution of I2 (1.50 g, 5.61 mmol) in MeOH (50 mL). Stir the resulting mixture at room temperature for 2 h. Then dilute the mixture with water (80 mL) and extract twice with MTBE (80 mL). Wash the combined organic layers with brine (100 mL), dry over anhydrous Na 2 SO 4 dry and concentrate to dryness. Purify the residue by silica gel column chromatography (eluting with PE:EtOAc = 100:0 to 40:1) to obtain I3 as a white solid (1.25 g, 84.60% yield).

[0346] Step 4: At room temperature, add CuSO 4 (8 mg, 0.05 mmol) and sodium ascorbate (77 mg, 0.39 mmol) to a mixture of 4-azidobenzonitrile (70 mg, 0.48 mmol) and I3 (140 mg, 0.53 mmol) in DMSO (6 mL). Stir the resulting mixture at 60 °C under N 2Stir for 3 hours under the atmosphere. Then dilute the mixture with water (30 mL) and extract twice with DCM (15 mL). Wash the combined organic layers with brine (20 mL), dry over anhydrous Na 2 SO 4 and filter, then concentrate to dryness. Purify the residue by silica gel column chromatography (eluting with PE:EtOAc = 100:0 to 4:1) to obtain Boc-I4 as a yellow oil (94 mg, 47.51% yield). LC / MS (ESI) m / z: 408 (M+H) + . At 0 °C, under N2 atmosphere, add 4N HCl / dioxane solution (5 mL) to a container containing Boc-I4 (94 mg, 0.23 mmol). Stir the resulting mixture at room temperature for 2 hours. Then concentrate the mixture under reduced pressure to obtain crude I4HCl as a pale yellow oil (75 mg, 95.07% yield), which is used directly in the next step without further purification. LC / MS (ESI) m / z: 308 (M+H) + .

[0347] Step 5: At 0 °C, add CDI (47 mg, 0.29 mmol) to a mixture of D1HCl (59 mg, 0.29 mmol) and DIPEA (95 mg, 0.73 mmol) in DMF (8 mL). Stir the mixture at room temperature for 1 hour. Then add I4HCl (75 mg, 0.23 mmol) to the above mixture, and stir the resulting mixture at 40 °C for another 3 hours. After cooling, quench the mixture with saturated NaHCO 3 aqueous solution (20 mL) and extract twice with EtOAc (15 mL). Wash the combined organic layers with saturated NH 4 Cl solution (30 mL) and brine (30 mL), dry over anhydrous Na 2 SO 4 and concentrate to dryness. Purify the crude product by silica gel column chromatography (DCM:MeOH = 100:0 to 15:1) to obtain Boc-I5 as an off-white solid (55 mg, 42.23% yield). LC / MS (ESI) m / z: 434 (M - 100 + H) + . At 0 °C, under N2 atmosphere, add 4N HCl / dioxane solution (5 mL) to Boc-I5 (55 mg, 0.10 mmol) in portions. Stir the resulting mixture at room temperature for 2 hours. Then concentrate the mixture to dryness under reduced pressure. Purify the residue by preparative HPLC to obtain I5 as a colorless oil (18 mg, 40.29% yield).

[0348]

[0349] Example 82: Example 82 was prepared according to the following method.

[0350]

[0351] To a solution of J1 (15 mg, 53 μmol) and 5-((tert-butoxycarbonyl)amino)pentanoic acid (15 mg, 69 μmol) in DMF (0.5 mL) was added DIPEA (28.5 μL, 106 μmol) and HATU (26.3 mg, 69 μmol). The solution was stirred at room temperature and then concentrated to a residue, which was taken up in 4N HCl / dioxane (1 mL). The solution was kept at room temperature and then concentrated to a residue. The product was taken up in methanol and then purified by preparative reverse-phase HPLC to give J2.

[0352]

[0353] Example 83: Example 83 was prepared according to the following method.

[0354]

[0355] Step 1: To a solution of (rac)-A2 (108 mg, 381 μmol) in THF (2 mL) was added 4-chlorobenzoic acid hydrazide (65.1 mg, 381 μmol), triethylamine (159 μL, 1.14 mmol) and T3P (587 mg, 953 μmol). The solution was heated at 75 °C for 3 h and then overnight at room temperature. The solution was poured into water and the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were washed twice with saturated aqueous sodium bicarbonate and then once with brine, dried over MgSO4, filtered and concentrated to a residue, which was purified by column chromatography to give H1 (117 mg, 270 μmol) as a white solid. LC / MS (ESI) m / z: 335.9 (M - 100 + H) + .

[0356] Step 2: To a solution of H1 (118 mg, 268 μmol) in MeCN (1 mL) was added DIPEA (94 μL, 547 μmol) and pTsCl (202 mg, 536 μmol). The solution was stirred at room temperature for three hours and then diluted with water and ethyl acetate. The phases were separated and the organic phase was washed with saturated aqueous NaHCO3, dried over MgSO4, filtered and concentrated. The product was purified by column chromatography to give H2 (85 mg, 203 μmol) as a colorless oil. LC / MS (ESI) m / z: 362.2 (M - 57 + H) + .

[0357] Step 3: Add HCl / dioxane (1 mL) to a vial containing H2 (85 mg, 203 umol). Keep the solution at room temperature for 30 minutes and then concentrate to give H3 as a colorless residue, which can be used without further purification.

[0358] Step 4a: At 0 °C, add E1 (974 mg, 9.72 mmol) to a suspension of CDI (2.11 g, 14.68 mmol) in DCM (10 mL). Stir the solution at room temperature for fifteen minutes and then concentrate to a residue and purify by column chromatography (0 - 100% DCM / methanol). Recover the product H4 (1.74 g) as a viscous oil.

[0359] Step 4b: Add H4 (44 mg, 225 umol) to a solution of H3 (40 mg, 113 umol) and DIPEA (40 uL, 225 umol) in DMF (0.5 mL). Heat the solution at 50 °C for 2 h and then cool to room temperature. Purify the reaction mixture directly by reverse-phase preparative HPLC to give H5 as a colorless oil.

[0360]

[0361]

[0362] Incorporation by reference

[0363] All U.S. patents and U.S. patent application publications cited herein are hereby incorporated by reference.

[0364] Equivalent embodiments

[0365] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the appended claims.

Claims

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

2. The compound according to claim 1, which has the following structure:

3. The compound according to claim 1 or 2, wherein L 1 is selected from -NH- and -N(CH 3 )-.

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

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

6. The compound according to claim 5, wherein R 1 is -H.

7. The compound according to claim 5, wherein R 1 is -NH 2 .

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

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

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

11. The compound according to claim 10, wherein R 2 and R 3 together with the carbon atom to which they are attached form an unsubstituted cyclopropyl or cyclobutyl group.

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

13. The compound according to claim 12, wherein R 2 and R 3 together with the carbon atom to which they are attached form an unsubstituted azetidinyl, pyrrolidinyl, piperidinyl or lactam.

14. The compound according to claim 12, wherein R 2 and R 3 together with the carbon atom to which they are attached form a substituted azetidinyl, pyrrolidinyl, piperidinyl or lactam.

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

16. The compound according to any one of claims 1-15, wherein X 1 is selected from 17. The compound according to any one of claims 1-16, wherein L 3 is triazolyl, oxazolyl or oxadiazolyl.

18. A compound according to any one of claims 1-17, wherein -L 3 -X 2 is selected from 19. The compound according to any one of claims 1-17, wherein -L 3 -X 2 is 20. The compound according to any one of claims 1-19, wherein X 2 is an unsubstituted aryl group.

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

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

23. The compound according to claim 22, wherein the substituted aryl is substituted phenyl.

24. The compound according to claim 23, wherein X 2 is and R 4 、R 5 、R 6 、R 7 and R 8 are independently selected from -H, halogen, -CN, -CF 3 、-CHF 2 、-OCF 3 、-OCHF 2 、alkyl, alkenyl, alkynyl and cycloalkyl; provided that at least one of R 4 、R 5 、R 6 、R 7 and R 8 is not -H.

25. The compound according to claim 24, wherein R 4 , R 5 , R 6 , R 7 and R 8 are independently selected from -H, -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl; provided that at least one of R 4 , R 5 , R 6 , R 7 and R 8 is not -H.

26. The compound according to claim 25, wherein X 2 is and R 4 selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

27. The compound according to claim 25, wherein X 2 is and R 5 selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

28. The compound according to claim 25, wherein X 2 is and R 6 selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

29. The compound according to claim 23, wherein X 2 is and R 5 and R 6 are independently selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

30. The compound according to claim 23, wherein X 2 is and R 4 and R 6 are independently selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

31. The compound according to claim 23, wherein X 2 is and R 5 、 R 6 and R 7 are independently selected from -Cl, -Br, -F, -CN, -CF 3 , -OCF 3 , -CH 3 and cyclopropyl.

32. The compound according to any one of claims 1-25, wherein X 2 is selected from:

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

34. A compound according to any one of claims 1-33, wherein X3 is -CF 3 .

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

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

37. The compound according to claim 35, which has the following structure:

38. The compound according to claim 35, which has the following structure:

39. The compound according to claim 35, which has the following structure:

40. The compound according to claim 36, which has the following structure:

41. The compound according to claim 36, which has the following structure:

42. The compound according to claim 36, which has the following structure:

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

44. A pharmaceutical composition, which comprises a compound according to any one of claims 1-43 and a pharmaceutically acceptable excipient.

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

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

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

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

49. A method for treating or preventing tyrosinemia (type I, II or III), which comprises administering to a subject in need thereof an effective amount of a compound according to any one of claims 1-43.

50. The method according to claim 49, wherein the compound reduces the systemic tyrosine level in the subject.

51. A method for treating or preventing non-ketotic hyperglycinemia, which comprises administering to a subject in need thereof an effective amount of a compound according to any one of claims 1-43.

52. The method according to claim 51, wherein the compound reduces the systemic glycine level in the subject.

53. A method for treating or preventing isovaleric academia, methylmalonic academia, propionic academia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorder or hyperammonemia, which comprises administering to a subject in need an effective amount of a compound as described in any one of claims 1-43.

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

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

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