Small molecule inhibitors of mammalian SLC6A19 function

Abstract

To provide compounds useful for treating or preventing a disease or disorder associated with abnormal levels of amino acids by modulation of SLC6A19 transport.SOLUTION: A compound of the following formula (I) or a pharmaceutically acceptable salt thereof is provided (where n is 1; L1 is -alkyl-; L2 is absent; L3 is -C(O)-; X1 and X2 are each independently H or cycloalkyl, provided that X1 and X2 are not both H; Y1 is substituted or unsubstituted aryl; Y2 is hydroxyalkyl or -NH(Y2'); Y2' is alkyl; Y3, Y4, Y5, and Y6 are each independently H or halide).SELECTED DRAWING: None

Description

[Technical Field]

[0001] (Not applicable) (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of priority to U.S. Provisional Patent Application Nos. 63 / 308,790, filed February 10, 2022, 63 / 292,815, filed December 22, 2021, 63 / 234,487, filed August 18, 2021, 63 / 226,551, filed July 28, 2021, and 63 / 159,271, filed March 10, 2021. [Background technology]

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

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

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

[0005] Accordingly, provided herein are compounds having the structure of formula (I): [ka] (In the formula, n is 0, 1, or 2; L1 is absent or selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH2-; L2 is absent or is -CH2-; L3 is absent or is -C(O)-; X1 and X2 are independently selected from -H, alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl, with the proviso that X1 and X2 are not both -H; Y1 is selected from aryl and heteroaryl; Y2 is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NH(Y2'), and -N(Y2'')2; Y2' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, and cycloalkyl; each Y2" is alkyl, or both occurrences together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocyclyl; Y3, Y4, Y5, and Y6 are independently selected from -H, -OH, halide, alkyl, haloalkyl, and alkoxy, with the proviso that Y3 and Y4 or Y5 and Y6 are not both -OH; provided that when L3 is -C(O)-, Y2 is not aryl and the compound is [ka] (not selected from or a pharmaceutically acceptable salt thereof.

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

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

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

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

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

[0011] [Figure 1-1] 1 is a table summarizing isoleucine transport data for exemplary compounds of the invention: A=IC50<500 nM, B=IC50 500 nM-1500 nM, C=IC50 1500 nM-5000 nM, D=IC50 5000 nM-10000 nM, E=IC50>10000 nM. [Figure 1-2] Continued from Figure 1-1. [Figure 1-3]Continued from Figure 1-2. [Figure 2] 1 is a table summarizing isoleucine transport data for additional exemplary compounds of the invention: A=IC50<500 nM, B=IC50 500 nM-1,500 nM, C=IC50 1,500 nM-5,000 nM, D=IC50 5,000 nM-10,000 nM, and E=IC50>10,000 nM. [Figure 3] 1 is a table summarizing isoleucine transport data for additional exemplary compounds of the invention: A=IC50<500 nM, B=IC50 500 nM-1,500 nM, C=IC50 1,500 nM-5,000 nM, D=IC50 5,000 nM-10,000 nM, and E=IC50>10,000 nM. DETAILED DESCRIPTION OF THE INVENTION

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

[0013] To facilitate a more readily understood understanding of the present invention, certain terms and phrases are defined below and throughout the specification.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0031] A "therapeutically effective amount" (or "effective amount") of a compound for use in therapy refers to the amount of compound in a preparation that, when administered (to a mammal, preferably a human) as part of a desired dosing regimen, alleviates the symptoms, improves the condition, or delays the onset of a disease state, in accordance with clinically accepted criteria for the disease or condition or cosmetic purpose being treated, e.g., at a reasonable benefit / risk ratio applicable to any medical treatment.

[0032] The term "prophylactic or therapeutic" treatment is art-recognized and includes administration to a host of one or more of the subject compositions. When performed prior to the clinical manifestation of an undesirable condition (e.g., a disease or other undesirable condition in a host animal), the treatment is prophylactic (i.e., protects the host from the occurrence of the undesirable condition), whereas when performed after the manifestation of the undesirable condition, the treatment is therapeutic (i.e., intended to reduce, ameliorate, or stabilize an existing undesirable condition or its side effects).

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

[0034] Aliphatic chains include the classes alkyl, alkenyl, and alkynyl, as defined below. A straight-chain aliphatic chain is limited to an 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 groups.

[0035] "Alkyl" refers to a fully saturated, cyclic or acyclic, branched or unbranched carbon chain moiety having the specified number of carbon atoms, or 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, as well as moieties that are positional isomers of these moieties. Alkyl having 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, and tetracosyl. In certain embodiments, a straight or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C6 for straight chain). 30 , C3-C for branched chain 30 ), more preferably having 20 or fewer carbon atoms. The alkyl group can be substituted or unsubstituted.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0059] Compounds of the Invention One aspect of the present invention is a compound of formula (I): [ka] (In the formula, n is 0, 1, or 2; L1 is absent or selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH2-; L2 is absent or is -CH2-; L3 is absent or is -C(O)-; X1 and X2 are independently selected from -H, alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl, with the proviso that X1 and X2 are not both -H; Y1 is selected from aryl and heteroaryl; Y2 is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NH(Y2'), and -N(Y2'')2; Y2' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, and cycloalkyl; each Y2" is alkyl, or both occurrences together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocyclyl; Y3, Y4, Y5, and Y6 are independently selected from -H, -OH, halide, alkyl, haloalkyl, and alkoxy, with the proviso that Y3 and Y4 or Y5 and Y6 are not both -OH; provided that when L3 is -C(O)-, Y2 is not aryl and the compound is [ka] (not selected from or a pharmaceutically acceptable salt thereof.

[0060] In certain embodiments, the compound also [ka] or a pharmaceutically acceptable salt thereof.

[0061] In certain embodiments, the compound has a structure selected from: [ka]

[0062] In certain embodiments, Y2' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, and cycloalkyl.

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

[0064] In certain embodiments, one of X1 and X2 is -H and the other of X1 and X2 is -CH3, -CH2CH3, -CH2CF3, -CH2CH2CH3, [ka] is selected from.

[0065] In certain embodiments, X1 is -H and X2 is -CH 3であるか、 X2 is -H and X1 is -CH3, or X1 is -H and X2 is [ka] or X2 is -H and X1 is [ka] is.

[0066] In certain embodiments, L1 is absent. In other embodiments, L1 is selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH2-.

[0067] In certain embodiments, L1 is selected from -CH2-, -C(H)(CH3)-, -CH2CH2-, and -C(H)(OH)CH2-. [ka] In other embodiments, L1 is [ka] In other embodiments, L1 is selected from: [ka] is selected from.

[0068] In certain embodiments, the compound is selected from: [ka]

[0069] In certain embodiments, Y 1 is unsubstituted aryl, such as unsubstituted phenyl and unsubstituted naphthyl.

[0070] In certain embodiments, Y 1 is substituted aryl.

[0071] In certain embodiments, Y is [ka] and R1, R2, R3, R4, and R5 are each independently -H, halogen, -CN, 、 Independently selected from -CF3, -CHF2, -CF2CH3, -OCF3, -OCHF2, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl, with the proviso that one of R1, R2, R3, R4, and R5 is not -H.

[0072] In certain embodiments, R1, R2, R3, R4, and R5 are -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH 3、 -CF3, -CHF2, -CF2CH 3、 -OCH3, -OCF3, -OCHF2, [ka] are independently selected from

[0073] In certain embodiments, R1, R2, R3, R4, and R5 are -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH 3、 -OCF3, and [ka] are independently selected from

[0074] In certain embodiments, two of R1, R2, R3, R4, and R5 are not -H. In other embodiments, three of R1, R2, R3, R4, and R5 are not -H.

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

[0076] In certain embodiments, Y is selected from: [ka] is selected from.

[0077] In certain embodiments, Y 1 is unsubstituted heteroaryl.

[0078] In certain embodiments, Y is [ka] is selected from.

[0079] In certain embodiments, Y 1 is substituted heteroaryl.

[0080] In certain embodiments, Y is [ka] is selected from Each occurrence of R6, R7, R8, and R9 is independently selected from -H, halogen, -CN, -OCF3, -OCHF2, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, aryl, and heteroaryl, provided that at least one of R6, R7, R8, and R9 is not -H.

[0081] In certain embodiments, L2 is absent. In other embodiments, L2 is -CH2-.

[0082] In certain embodiments, L3 is absent. In other embodiments, L3 is -C(O)-.

[0083] In certain embodiments, n is 0. In other embodiments, n is 1. In other embodiments, n is 2.

[0084] In certain embodiments, the compound is selected from: [ka]

[0085] In certain embodiments, the compound is selected from: [ka]

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

[0087] In certain embodiments, Y2 is unsubstituted heteroaryl.

[0088] In certain embodiments, Y2 is [ka] is selected from.

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

[0090] In certain embodiments, Y2 is substituted heteroaryl.

[0091] In certain embodiments, Y2 is [ka] and R 10 , R 11 , and R 12 -H, halogen, -CN, -OH 、-NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 -CO2R 15 , and -C(O)NHSO2R 15 are independently selected from, where R 10 , R 11 , and R 12 At least one of the is not -H, R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0092] In certain embodiments, Y2 is [ka] and R 10 , R 11 , and R 12 -H, halogen, -CN, -OH 、 -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 -CO2R 15 , and -C(O)NHSO2R 15 are independently selected from, where R 10 , R 11 , and R 12 At least one of the is not -H, R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0093] In certain embodiments, R 10 , R11 , and R 12 -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH 3、 -CF3, -CHF2, -CF2CH 3、 independently selected from —OCH3, —OCF3, —OCHF2, —OAc, —NH2, —NHCH3, —NHAc, —C(O)NH2, —C(O)NHCH3, —C(O)NHCH2CH3, —C(O)NHSO2CH3, —C(O)NHSO2CH2CH3, —CO2H, phenyl, cyclopropyl, cyclobutyl, imidazolyl, and tetrazolyl.

[0094] In certain embodiments, R 10 and R 12 are -H and R, respectively. 11 is -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, [ka] is selected from.

[0095] In certain embodiments, R 11 and R 12 are -H and R, respectively 10 -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, [ka] is selected from.

[0096] In certain embodiments, R 10 and R 11 are -H and R, respectively 12 -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, [ka] is selected from.

[0097] In certain embodiments, Y2 is [ka] is selected from R 16 is, for each occurrence, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14 , -CO2R 15 are independently selected from R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0098] In certain embodiments, R 16 are -CN, -CH3, -CF3, -C(O)NH2, -CO2CH2CH3, and [ka] is selected from.

[0099] In certain embodiments, Y2 is [ka] is selected from R 17 , R 18 , R 19 , R 20 , and R 21Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from, where R 17 , R 18 , R 19 , R 20 , and R 21 At least one of the is not -H, R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0100] In certain embodiments, Y2 is [ka] is selected from R 17 , R 18 , R 19 , R 20 , and R 21 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from, where R 17 , R 18 , R 19 , R 20 , and R 21 At least one of the is not -H, R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0101] In certain embodiments, R 17 , R 18 , R 19 , R 20 , and R 21 are independently selected from —H, —CN, —CH 3 , and —OCH 3 .

[0102] In certain embodiments, Y2 is [ka] is selected from.

[0103] In certain embodiments, the compound is selected from: [ka]

[0104] In certain embodiments, the compound is selected from: [ka]

[0105] In certain embodiments, the compound is selected from: [ka]

[0106] In certain embodiments, Y2 is unsubstituted cycloalkyl or heterocyclyl.

[0107] In certain embodiments, Y2 is [ka] is selected from.

[0108] In certain embodiments, Y2 is [ka] is selected from.

[0109] In certain embodiments, Y2 is substituted cycloalkyl or heterocyclyl.

[0110] In certain embodiments, Y2 is [ka] is selected from.

[0111] In certain embodiments, Y2 is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, and hydroxyalkyl.

[0112] In certain embodiments, Y2 is selected from -CH3, -CH2CH(CH3)2, -CH2CH2C≡CH, -CH2CH2OCH3, -C(H)(CH3)CH2OCH3, -OCH3, -CH2OH, -CH2CH2OH, -C(CH3)2OH, and -CH2OCH3.

[0113] In certain embodiments, Y2 is selected from -CH2OH and -CH2CH2OH.

[0114] In certain embodiments, Y2 is heteroaryl.

[0115] In certain embodiments, Y2 is [ka] is selected from.

[0116] In certain embodiments, Y2 is [ka] and R 10 , R 11 , and R 12 -H, halogen, -CN, -OH、 -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0117] In certain embodiments, Y2 is [ka] and R 10 , R 11 , and R 12 -H, halogen, -CN, -OH 、 -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0118] In certain embodiments, R 10 , R 11 , and R 12 At least one of is not -H.

[0119] In certain embodiments, Y2 is [ka] is selected from R 17 , R 18 , R 19 , R 20 , and R 21 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0120] In certain embodiments, Y2 is [ka] is selected from R 17 , R 18 , R 19 , R 20 , and R 21 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0121] In certain embodiments, R 17 , R 18 , R 19 , R 20 , and R 21At least one of is not -H.

[0122] In certain embodiments, Y2 is [ka] is selected from R 22 , R 23 , R 24 , and R 25 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 Each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0123] In certain embodiments, R 22 , R 23 , R 24 , and R 25 Each occurrence of is independently selected from -H, and -CH3.

[0124] In certain embodiments, Y2 is -NH(Y2'), or Y2 is -N(Y2'')2.

[0125] In certain embodiments, Y2' is -H, -OH, -OCH3, -CH3, -CH2CH2OCH3, and [ka] is selected from.

[0126] In certain embodiments, each Y2" is -CH3. In other embodiments, both Y2" together with the nitrogen atom to which they are attached form morpholinyl.

[0127] In certain embodiments, Y2 is -NH(Y2').

[0128] In certain embodiments, Y2' is selected from -H, alkyl, alkoxy, and hydroxyalkyl. In other embodiments, Y2' is selected from -H, -OCH3, -CH3, and -CH2CH2OH.

[0129] In certain embodiments, Y2' is H. In other embodiments, Y2' is -CH3. In other embodiments, Y2' is -CH2CH3. In other embodiments, Y2' is -OCH3. In other embodiments, Y2' is -CH2OH. In other embodiments, Y2' is -CH2CH2OH.

[0130] In certain embodiments, Y and Y are both -H or -F. In other embodiments, Y is selected from -F, -CF, -OH, and -OCH, and Y is -H. In other embodiments, Y is selected from -F, -CF, -OH, and -OCH, and Y is -H.

[0131] In certain embodiments, Y3 and Y4 are both -H. In other embodiments, Y3 and Y4 are both -F.

[0132] In certain embodiments, Y5 and Y6 are both -H or -F. In other embodiments, Y5 is selected from -F, -CF3, -OH, and -OCH3, and Y6 is -H. In other embodiments, Y6 is selected from -F, -CF3, -OH, and -OCH3, and Y5 is -H.

[0133] In certain embodiments, Y5 and Y6 are both -H. In other embodiments, Y5 and Y6 are both -F.

[0134] In some embodiments, the compound is selected from Table 1 below: [Table 1] TIFF0007797723000050.tif250157 TIFF0007797723000051.tif250156 TIFF0007797723000052.tif245167 TIFF0007797723000053.tif250159 TIFF0007797723000054.tif225167 TIFF0007797723000055.tif250158 TIFF0007797723000056.tif250162 TIFF0007797723000057.tif249167 TIFF0007797723000058.tif238167 TIFF0007797723000059.tif250159 TIFF0007797723000060.tif250167 TIFF0007797723000061.tif250158 TIFF0007797723000062.tif210167 TIFF0007797723000063.tif250157 TIFF0007797723000064.tif246167 TIFF0007797723000065.tif251167 TIFF0007797723000066.tif250167 TIFF0007797723000067.tif252167 TIFF0007797723000068.tif250161 TIFF0007797723000069.tif251167

[0135] In some embodiments, the compound is selected from Table 2 below: [Table 2] TIFF0007797723000071.tif211167 TIFF0007797723000072.tif240167 TIFF0007797723000073.tif225167 TIFF0007797723000074.tif250167 TIFF0007797723000075.tif243167 TIFF0007797723000076.tif250163 TIFF0007797723000077.tif246167 TIFF0007797723000078.tif233167

[0136] In some embodiments, the compound is selected from Table 3 below: [Table 3] TIFF0007797723000080.tif243167 TIFF0007797723000081.tif237167 TIFF0007797723000082.tif114167

[0137] In some embodiments, the compound is selected from Table 4 below: [Table 4]

[0138] In some embodiments, the compound is selected from Table 5 below: [Table 5] TIFF0007797723000085.tif250162 TIFF0007797723000086.tif246167

[0139] In some embodiments, the compounds are atropisomers. Additionally, unless otherwise stated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, the replacement of a hydrogen by deuterium or tritium, or 13 C or 14 Compounds produced by the replacement of carbon with C-enriched carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or therapeutic agents according to the present invention. For example, the variable group R 1 In the case of -C1-C4, the (C1-C4) alkyl or -O-(C1-C4) alkyl can be suitably deuterated (e.g., -CD3, -OCD3).

[0140] Any of the compounds of the present invention may also be radiolabeled for the preparation of radiopharmaceuticals.

[0141] Treatment method One aspect of the present invention provides compounds, compositions, and methods useful for treating or preventing diseases or disorders associated with abnormal amino acid concentrations through modulation of SLC6A19 transport.

[0142] Another aspect of the present invention relates to a method of modulating SLC6A19 transport in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0143] Another aspect of the present invention relates to a method of treating or preventing a disease or disorder associated with a genetic deficiency in phenylalanine hydroxylase in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0144] In some embodiments, the present invention relates to a method of treating or preventing phenylketonuria in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0145] In some embodiments, the present invention relates to a method of treating or preventing hyperphenylalaninemia in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0146] In some embodiments, the compound decreases phenylalanine levels in the body of a subject.

[0147] In some embodiments, the present invention relates to a method of treating or preventing tyrosinemia (Type I, II, or III) in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0148] In some embodiments, the compound decreases glycine levels in the subject's body.

[0149] In some embodiments, the present invention relates to a method of treating or preventing isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, a urea cycle disorder, or hyperammonemia in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I).

[0150] In some embodiments of any one of the disclosed methods, the compound modulates SLC6A19 in the subject.

[0151] In some embodiments of any one of the disclosed methods, the compound inhibits SLC6A19 in the subject.

[0152] In some embodiments of any one of the disclosed methods, the compound modulates SLC6A19 transport in the subject.

[0153] In some embodiments of any one of the disclosed methods, the compound inhibits SLC6A19 transport in the subject.

[0154] In some embodiments, the compound decreases amino acid concentrations in the subject's body.

[0155] In some embodiments of any one of the disclosed methods, the subject is a mammal. In some embodiments of any one of the disclosed methods, the mammal is a human.

[0156] In some embodiments of any one of the disclosed methods, the compound of formula (I) is defined as follows: [ka] (I) (In the formula, n is 0, 1, or 2; L1 is absent or selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH2-; L2 is absent or is -CH2-; L3 is absent or is -C(O)-; X1 and X2 are independently selected from -H, alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl, with the proviso that X1 and X2 are not both -H; Y1 is selected from aryl and heteroaryl; Y2 is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, hydroxyalkyl, aralkyl, hetaralkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NH(Y2'), and -N(Y2'')2; Y2' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, and cycloalkyl; each Y2" is alkyl, or both occurrences together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocyclyl; Y3, Y4, Y5, and Y6 are independently selected from -H, -OH, halide, alkyl, haloalkyl, and alkoxy, with the proviso that Y3 and Y4 or Y5 and Y6 are not both -OH. or a pharmaceutically acceptable salt thereof.

[0157] In some embodiments of any one of the disclosed methods, the compound is selected from: [ka] or a pharmaceutically acceptable salt thereof.

[0158] In some embodiments of any one of the disclosed methods, the compound is selected from: [ka] or a pharmaceutically acceptable salt thereof.

[0159] In some embodiments of any one of the disclosed methods, the compound is selected from the structure of any one of the compounds listed in Table 1.

[0160] In some embodiments of any one of the disclosed methods, the compound is selected from the structure of any one of the compounds listed in Table 2.

[0161] In some embodiments of any one of the disclosed methods, the compound is selected from the structure of any one of the compounds listed in Table 3.

[0162] In some embodiments of any one of the disclosed methods, the compound is selected from the structure of any one of the compounds listed in Table 4.

[0163] In some embodiments of any one of the disclosed methods, the compound is selected from the structure of any one of the compounds listed in Table 5.

[0164] Pharmaceutical Compositions, Routes of Administration, and Dosing In certain embodiments, the present invention is directed to a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier. In certain embodiments, a pharmaceutical composition comprises multiple compounds of the present invention and a pharmaceutically acceptable carrier.

[0165] In certain embodiments, the pharmaceutical compositions of the present invention further comprise at least one additional pharmaceutically active agent other than the compound of the present invention. The at least one additional pharmaceutically active agent may be an agent useful in the treatment of ischemia-reperfusion injury.

[0166] Pharmaceutical compositions of the invention can be prepared by combining one or more compounds of the invention with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents.

[0167] As stated above, "effective amount" refers to any amount sufficient to achieve a desired biological effect. In conjunction with the teachings provided herein, an effective prophylactic or therapeutic treatment regimen can be designed by selecting from among various active compounds and considering factors such as potency, relative bioavailability, patient weight, severity of adverse side effects, and mode of administration to effectively treat a particular subject without causing substantial unnecessary toxicity. The effective amount for any particular application may vary depending on factors such as the disease or condition being treated, the particular compound of the present invention being administered, the subject's size, or the severity of the disease or condition. Those skilled in the art can empirically determine the effective amount of a particular compound of the present invention and / or other therapeutic agent without necessitating undue experimentation. A maximum dose, i.e., the maximum safe dose according to medical judgment, may be used. Multiple administrations per day may be contemplated to achieve an appropriate internal concentration of the compound. An appropriate internal concentration can be determined, for example, by measuring the peak or maintenance concentration of the drug in the patient's plasma. "Dose" and "administration" are used interchangeably herein.

[0168] In certain embodiments, intravenous administration of the compound may typically be 0.1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 0.1 mg / kg / day to 2 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 0.5 mg / kg / day to 5 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be 1 mg / kg / day to 10 mg / kg / day.

[0169] Generally, the daily oral dose of the compound for human subjects is about 0.01 mg / kg / day to 1000 mg / kg / day. Oral doses in the range of 0.5 to 50 mg / kg, administered one or more times per day, are expected to produce therapeutic effects. The dosage can be appropriately adjusted to achieve the desired local or systemic drug concentration, depending on the mode of administration. For example, with intravenous administration, the daily dose is expected to be one to several orders of magnitude lower. If the subject does not respond adequately to such doses, higher doses (or effective high doses via another, more localized delivery route) can be employed, as tolerated by the patient. Multiple daily administrations are contemplated to achieve adequate internal concentrations of the compound.

[0170] For any compound described herein, the therapeutically effective amount can be first determined from animal models.The therapeutically effective amount can also be determined from human data for compounds tested in humans and for compounds known to exhibit similar pharmacological activity, such as other related active agents.Higher doses may be required for parenteral administration.The applied dose can be adjusted according to the relative bioavailability and efficacy of the administered compound.Adjusting the dose to achieve maximum efficacy based on the methods described above and other methods well known in the art is well within the capabilities of those skilled in the art.

[0171] The formulations of the present invention may be administered in pharmaceutically acceptable solutions, which may conventionally contain pharmaceutically acceptable concentrations of salts, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.

[0172] When used in therapy, an effective amount of compound can be administered to a subject by any method that delivers the compound to the desired surface.The administration of pharmaceutical compositions can be carried out by any means known to those skilled in the art.Administration routes include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (bladder), oral, subcutaneous, direct injection (e.g., into tumor or abscess), mucosal (e.g., topical to the eye), inhalation, and topical.

[0173] For intravenous and other parenteral administration routes, the compounds of the present invention can be formulated as lyophilized preparations, as lyophilized preparations of liposome intercalated or liposome-encapsulated active compounds, as lipid complexes in aqueous suspension, or as salt complexes. Lyophilized preparations are generally reconstituted with a suitable aqueous solution, such as sterile water or physiological saline, immediately prior to administration.

[0174] For oral administration, compounds can be easily formulated by combining the active compound(s) with pharmaceutically acceptable carriers well known in the art. Such carriers allow the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by the treated subject. Pharmaceutical preparations for oral use can be obtained as solid excipients, optionally by grinding the resulting mixture, and optionally adding suitable excipients, followed by processing the granular mixture to obtain tablets or dragee cores. Suitable excipients are, in particular, sugars, including fillers such as lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, e.g., sodium alginate. Optionally, oral preparations may also be formulated with saline or buffers, e.g., EDTA, for neutralizing acidic conditions in the body, or may be administered without any carrier.

[0175] Oral dosage forms of one or more of the components described above are also specifically contemplated. One or more of the components may be chemically modified to facilitate oral delivery of the derivatives. Generally, contemplated chemical modifications involve attaching at least one moiety to the component molecule itself that (a) inhibits acid hydrolysis and (b) allows uptake from the stomach or intestine into the bloodstream. It may also be desirable to increase the overall stability of one or more of the components and extend their circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, and polyproline. Abuchowski and Davis, "Soluble Polymer-Enzyme Adducts," In: Enzymes as Drugs, Hochenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367-383 (1981); Newmark et al., J. Appl. Biochem. 4:185-9 (1982). Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane. For pharmaceutical applications, as noted above, polyethylene glycol moieties are preferred.

[0176] The location of release of the component (or derivative) may be the stomach, the small intestine (duodenum, jejunum, or ileum), or the large intestine. One skilled in the art has available formulations that will not dissolve in the stomach but will release the substance in the duodenum or elsewhere in the intestine. Preferably, the release will avoid adverse effects in the stomach environment, either by protecting the compound (or derivative) of the invention or by releasing the biologically active substance in the intestine, etc., after passing through the stomach environment.

[0177] To ensure full gastric resistance, a coating that is impermeable to at least pH 5.0 is essential. Examples of more common inactive ingredients used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can also be used as mixed films.

[0178] A coating or mixture of coatings can also be used on tablets not intended for gastric protection. This can include sugar coatings or coatings that make the tablet easier to swallow. Capsules can consist of a hard shell (such as gelatin) for delivery of dry therapeutics (e.g., powder), or a soft gelatin shell can be used for liquid forms. The shell material for cachets can be thick starch or other edible paper. For pills, lozenges, molded tablets, or powder tablets, wet massing techniques can be used.

[0179] The therapeutic agent may also be included in the formulation as fine multiparticulates in the form of granules or pellets about 1 mm in size. The formulation of material for capsule administration may also be as a powder, lightly compressed plugs, or tablets. The therapeutic agent may be prepared by compression.

[0180] Both colorants and flavoring agents may be included. For example, the compounds (or derivatives) of the present invention may be formulated (such as encapsulated in liposomes or microspheres) and then further included in an edible product, such as a refrigerated beverage, containing colorants and flavoring agents.

[0181] The volume of the therapeutic agent can be diluted or increased with an inert material. These diluents can include carbohydrates, especially mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans, and starch. Certain inorganic salts can be used as fillers, including calcium triphosphate, magnesium carbonate, and sodium chloride. Some commercially available diluents include Fast-Flo, Emdex, STA-Rx 1500, Emcompress, and Avicell.

[0182] Disintegrants may be included in the formulation of therapeutic agents to form solid dosage forms. Materials used as disintegrants include, but are not limited to, starch, including the commercially available starch-based disintegrant Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethylcellulose, sponge, and bentonite may also be used. Another form of disintegrant is an insoluble cationic exchange resin. Powdered gums can be used as binders, including powdered gums such as agar, Karaya, or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

[0183] Binders can be used to hold the therapeutic agent and form a hard tablet and include materials derived from natural products such as gum arabic, tragacanth, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can both be used in alcoholic solution to granulate the therapeutic agent.

[0184] Antifriction agents may be included in the formulation of the therapeutic agent to prevent sticking during the formulation process. Lubricants can be used as a layer between the therapeutic agent and the die wall, and include, but are not limited to, stearic acid with magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, and Carbowax 4000 and 6000 can also be used.

[0185] Glidants may be added which may improve the flowability of the drug during formulation and aid in rearrangement during compression. Glidants may include starch, talc, pyrogenic silica, and hydrated silicoaluminate.

[0186] Surfactants may be added as wetting agents to aid in the dissolution of therapeutic agents in aqueous environments. Surfactants may include anionic surfactants such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate, and dioctyl sodium sulfonate. Cationic surfactants may also be used, including benzalkonium chloride and benzethonium chloride. Potential nonionic surfactants that can be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose. These surfactants may be present alone or in a mixture of different ratios in the formulation of the compound or derivative of the present invention.

[0187] Orally available pharmaceutical preparations include push-fit capsules made of gelatin as well as sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient mixed with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optional stabilizers. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid such as fatty oils, liquid paraffin, or liquid polyethylene glycol. Additionally, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All oral formulations must be in a dosage suitable for such administration.

[0188] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0189] For topical administration, the compounds may be formulated as solutions, gels, ointments, creams, suspensions, etc., as is well known in the art. Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, and those designed for transdermal, transmucosal, oral or pulmonary administration.

[0190] For administration by inhalation, the compound for use according to the present invention can be conveniently delivered in the form of aerosol spray presentation from a pressurized pack or nebulizer by using suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.For pressurized aerosol, dosage unit can be determined by providing a valve that delivers a metered amount.For example, gelatin capsules and cartridges for use in inhaler or insufflator can be formulated to contain a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0191] Also contemplated herein is pulmonary delivery of the compounds disclosed herein (or salts thereof). The compounds are delivered to the lungs of a mammal during inhalation, cross the epithelial lining of the lungs, and reach the bloodstream. Other reports on inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprorelin acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl. 5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (α1-proteinase); Oswein et al., 1990, "Aerosolization of Proteins," Proceedings of Symposium on Respiratory Drug Delivery. II, Keystone, Colorado, March (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor-alpha) and U.S. Patent No. 5,284,656 to Platz et al. (granulocyte colony-stimulating factor; incorporated by reference). Methods and compositions for pulmonary delivery of systemically acting drugs are described in U.S. Patent No. 5,451,569, issued September 19, 1995 to Wong et al. (incorporated by reference).

[0192] Contemplated for use in the practice of the present invention are a variety of mechanical devices designed for pulmonary delivery of therapeutic products, including, but not limited to, nebulizers, metered dose inhalers, and dry powder inhalers, all of which are well known to those skilled in the art.

[0193] 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.

[0194] All such devices require the use of formulations suitable for dispensing and administering the compounds of the present invention. Typically, each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material in addition to the usual diluents, adjuvants, and / or carriers useful in therapeutics. The use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers is also contemplated. The chemically modified compounds of the present invention can be prepared into various formulations depending on the type of chemical modification or the type of device employed.

[0195] Formulations suitable for use in either jet or ultrasonic nebulizers typically contain a compound (or derivative) of the invention dissolved in water at a concentration of about 0.1 to 25 mg of biologically active compound of the invention per mL of solution. The formulation may also contain a buffer and a simple sugar (e.g., for inhibitor stabilization and to control osmolality). Nebulizer formulations may also contain a surfactant to reduce or prevent surface-induced aggregation of the compound of the invention caused by atomization of the solution to form the aerosol.

[0196] Formulations for use with metered dose inhalers generally comprise a fine powder containing the compound (or derivative) of the present invention suspended in a propellant with the aid of a surfactant. The propellant may be any conventional material employed for this purpose, such as a chlorofluorocarbon, hydrochlorofluorocarbon, hydrofluorocarbon, or hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or a combination thereof. Suitable surfactants include sorbitan trioleate and soybean lecithin. Oleic acid may also be useful as a surfactant.

[0197] Formulations for dispensing from powder inhalation devices comprise a finely divided dry powder containing a compound (or derivative) of the invention, and may contain a bulking agent such as lactose, sorbitol, sucrose, or mannitol in an amount sufficient to facilitate dispersion of the powder from the device, e.g., 50-90% by weight of the formulation. The compound (or derivative) of the invention should advantageously be prepared in particulate form having an average particle size of less than 10 micrometers (μm), most preferably 0.5-5 μm, for most effective delivery to the deep lung.

[0198] Nasal delivery of the pharmaceutical compositions of the present invention is also contemplated. Nasal delivery allows the pharmaceutical compositions of the present invention to enter the bloodstream directly after administration of the therapeutic product to the nose, without the product being deposited in the lungs. Nasal delivery formulations include those based on dextran or cyclodextran.

[0199] For nasal administration, a useful device is a small, hard bottle equipped with a metered-dose sprayer. In one embodiment, the metered dose is delivered by drawing a solution of the pharmaceutical composition of the present invention into a chamber of a fixed volume, which has an opening dimensioned to aerosolize the aerosol formulation by forming a spray when the liquid in the chamber is compressed. The chamber is compressed to administer the pharmaceutical composition of the present invention. In a specific embodiment, the chamber is a piston arrangement. Such devices are commercially available.

[0200] Alternatively, it is a plastic squeeze bottle with an opening or aperture sized to aerosolize the aerosol formulation by forming a spray when squeezed. The opening is usually in the top of the bottle, which is generally tapered to partially fit into the nasal passages for efficient administration of the aerosol formulation. Preferably, the nasal inhaler provides a metered amount of the aerosol formulation to administer a measured amount of drug.

[0201] When systemic delivery is desired, the compound can be formulated for parenteral administration by injection, for example, bolus injection or continuous infusion. Injectable preparations can be provided in unit dosage form, for example, in ampoules or multi-dose containers, with preservatives added. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and can contain formulatory agents such as suspending agents, stabilizers, and / or dispersing agents.

[0202] Pharmaceutical preparations for parenteral administration include aqueous solutions of the active compound in water-soluble form.Furthermore, suspensions of the active compound can be prepared as appropriate oily injection suspensions.Suitable lipophilic solutions or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions can contain substances that increase the viscosity of 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, allowing for the preparation of highly concentrated solutions.

[0203] Alternatively, the active compound may be in powder form for reconstitution with a suitable vehicle, eg, sterile pyrogen-free water, before use.

[0204] The compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.

[0205] In addition to the formulations described above, the compounds can also be formulated as depot preparations. Such long-acting preparations can be formulated using suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as sparingly soluble salts.

[0206] The pharmaceutical compositions may also comprise suitable solid- or gel-phase carriers or excipients, examples of which include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0207] Suitable liquid or solid pharmaceutical preparation forms include, for example, aqueous or saline solutions for inhalation, microencapsulation, cochleation, coating on fine gold particles, encapsulation in liposomes, nebulization, aerosolization, pellets for skin implantation, or dried on sharp objects for rubbing on the skin. Pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or sustained-release preparations of active compounds, which may be prepared using conventional excipients and additives and / or auxiliaries, such as disintegrants, binders, coating agents, swelling agents, lubricants, flavoring agents, sweeteners, or solubilizers, as described above. Pharmaceutical compositions are suitable for use in various drug delivery systems. For a brief review of drug delivery methods, see Langer R, Science 249:1527-33 (1990).

[0208] The compounds of the present invention, and optional other therapeutic agents, may be administered as is (neat) or in the form of a pharmaceutically acceptable salt or cocrystal. When used in medicine, the salt or cocrystal must be pharmaceutically acceptable; however, pharmaceutically unacceptable salts or cocrystals may conveniently be used to prepare the pharmaceutically acceptable salt or cocrystal. Such salts include, but are not limited to, those prepared from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, p-toluenesulfonic acid, tartaric acid, citric acid, methanesulfonic acid, formic acid, malonic acid, succinic acid, naphthalene-2-sulfonic acid, and benzenesulfonic acid. Additionally, such salts may be prepared as alkali metal or alkaline earth metal salts, such as sodium, potassium, or calcium salts of the carboxylic acid group.

[0209] 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).

[0210] The pharmaceutical compositions of the present invention contain an effective amount of a compound described herein and an optional therapeutic agent contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents, or encapsulating substances suitable for administration to humans or other vertebrates. The term "carrier" refers to a natural or synthetic organic or inorganic component with which an active ingredient is combined to facilitate application. The components of the pharmaceutical compositions also can be mixed with the compounds of the present invention and with each other in a manner such that there is no interaction that would substantially impair the desired pharmaceutical effect.

[0211] The therapeutic agent(s), including but not limited to, the compounds of the present invention, may be provided in particles. As used herein, particle refers to nanoparticles or microparticles (or larger particles in some cases) that may comprise all or part of the compounds of the present invention or other therapeutic agent(s) described herein. The particles may contain the therapeutic agent(s) in a core surrounded by a coating, including but not limited to, an enteric coating. The therapeutic agent(s) may also be dispersed throughout the particle. The therapeutic agent(s) may also be adsorbed onto the particle. The particles may have any order of release rate, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. In addition to the therapeutic agent(s), the particles may contain any material commonly used in the pharmaceutical and medical arts, including but not limited to, disintegrating, non-disintegrating, biodegradable, or non-biodegradable materials, or combinations thereof. The particles may be microcapsules containing the compounds of the present invention in solution or in a semi-solid state. The particles may be of virtually any shape.

[0212] Both non-biodegradable and biodegradable polymeric materials can be used to manufacture particles for delivering therapeutic agent(s). Such polymers can be natural or synthetic. The polymer is selected based on the desired period of release. Bioadhesive polymers of particular interest include the biodegradable hydrogels described in Sawhney HS et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein by reference. These include polyhyaluronic acid, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0213] The therapeutic agent(s) may be contained in a controlled-release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation is controlled. This refers to immediate-release and non-immediate-release formulations, where non-immediate-release formulations include, but are not limited to, sustained-release and delayed-release formulations. The term "sustained-release" (also called "extended-release") is used in its conventional sense to refer to a drug formulation that provides a sustained release of drug over an extended period of time, preferably, but not necessarily, providing a substantially constant blood concentration of drug over an extended period of time. The term "delayed-release" is used in its conventional sense to refer to a drug formulation in which there is a time lag between administration of the formulation and the release of drug from the formulation. "Delayed-release" may or may not involve a sustained release of drug over an extended period of time, and therefore may or may not be "sustained-release."

[0214] For the treatment of chronic conditions, the use of long-term sustained-release implants may be particularly suitable. "Long-term" release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and preferably 30 to 60 days. Long-term sustained-release implants are well known to those skilled in the art and include some of the release systems described above.

[0215] It will be understood by those skilled in the relevant art that other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the description of the invention contained herein, in view of the information known to those skilled in the art, and can be made without departing from the scope of the invention or any embodiment thereof. Having now described the invention in detail, the present invention will be more clearly understood by reference to the following examples. The examples are included herein for illustrative purposes only and are not intended to limit the invention. [Example]

[0216] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0217] Example 1: SLC6A19 isoleucine transport assay Cell line generation and maintenance The Flp-In™ T-REx™ 293 cell line was purchased from Thermo Fisher Scientific. This line was used to generate stable cell lines that inducibly express human SLC6A19 containing a C-terminal V5 tag and stably express human TMEM27 (also known as Collectrin) containing a C-terminal myc-DDK tag. Stable cell lines were generated by transfecting plasmids encoding SLC6A19 and TMEM27 using standard protocols, followed by antibiotic selection. Stable cells were maintained in DMEM / F12 supplemented with Glutamax, 10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin, 200 μg / mL hygromycin, 10 μg / mL blasticidin, and 300 μg / mL neomycin (Thermo Fisher).

[0218] Assay: Isoleucine transport assay in 96-well format On day 0, stable cell lines were seeded at a density of 35,000 cells / well in poly-D-lysine-coated, 96-well, cell culture-treated plates. On day 1, SLC6A19 expression was induced by dispensing tetracycline at a final concentration of 1 μg / mL using a Tecan D300e digital dispenser. On day 2, transport assays were performed. The medium was removed from the plate using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio), and the cells were washed with 175 μL of live cell imaging solution (Thermo Fisher Scientific) using the Blue Washer. After washing, cells were treated with either 70 μL of DMSO, positive control, or compound diluted in Krebs buffer (140 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgCl2, 11 mM HEPES, 10 mM glucose, pH 7.4) at room temperature. After 20–60 minutes, 30 μL of a 3.3 mM solution of 13C6,15N-L-isoleucine (Cambridge Isotope Laboratories) was added. After 20 minutes of incubation with the isoleucine substrate at room temperature, the cells were washed with 175 μL of live cell imaging solution using a Blue Washer. Cells were then lysed with 150 μL of 15 μM D-Leucine-d10 in ultrapure water (CDN Isotopes). To facilitate lysis, the plate was shaken at 700 rpm for a minimum of 40 minutes. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the untreated cell lysate. The plate was returned to the shaker for a minimum of 2 minutes to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000 rpm for 5 minutes to pellet cell debris and precipitate. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0219] Assay: Isoleucine transport assay in 384-well format On day 0, stable cell lines were seeded at a density of 20,000 cells / well into poly-D-lysine-coated, 384-well cell culture-treated plates containing 1 μg / mL tetracycline using a Viaflo 384-well pipette. Transport assays were performed the following day (day 1). The medium was removed from the plates using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio), and the cells were washed with 80 μL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, cells were treated with either 20 μL of DMSO, positive control, or compound diluted in Krebs buffer (140 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgCl2, 11 mM HEPES, 10 mM glucose, pH 7.4) using a TECAN liquid handler. After incubation at room temperature for 20-60 minutes, 8.6 μL of a 3.3 mM solution of 13C6,15N-L-isoleucine (Cambridge Isotope Laboratories) was added. After incubation with the isoleucine substrate for 20 minutes at room temperature, the cells were washed with 80 μL of live cell imaging solution using a Blue Washer. Cells were then lysed with 80 μL of 15 μM D-Leucine-d10 in ultrapure water (CDN Isotopes). To facilitate lysis, the plate was shaken at 700 rpm for a minimum of 2 hours. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the untreated cell lysate. The plate was returned to the shaker for a minimum of 5 minutes to ensure proper mixing of the standard curve. The plate was then centrifuged at 4,000 rpm for 10 minutes to pellet cell debris and precipitate. The supernatant was diluted 1:10 in acetonitrile + 0.1% formic acid in a polypropylene plate.

[0220] Analysis of 13C6,15N-L-isoleucine was performed using a RapidFire365-QTOF 6545 (Agilent). Quantitative sample analysis utilized automated solid-phase extraction (HILIC H6 cartridge) followed by mass spectrometric injection. Samples were loaded using 95% acetonitrile, 0.1% formic acid and eluted directly from the cartridge using 5% acetonitrile, 0.1% formic acid for ESI-MS (electrospray ionization). Analyte quantification was performed using Agilent Masshunter Quant software from high-resolution full-scan data.

[0221] General Procedure A: [ka] Step 1: Synthesis of Compound A2 To a solution of compound A1 (1 equivalent) in toluene was added a solution of bistrichloromethyl carbonate (BTC) (0.5 equivalents) in toluene dropwise at 0° C. under a N atmosphere. The resulting mixture was stirred at room temperature for 15 minutes, and then the mixture was heated to 130° C. and stirred under a N atmosphere for 2 hours. After cooling, the mixture was concentrated to dryness under reduced pressure to give crude compound A2, which was used directly in the next step without further purification.

[0222] Step 2: Synthesis of compound A4 To a mixture of compound A3 (1 equivalent) and TEA (3 equivalents) in anhydrous DCM was added a solution of compound A2 (1 equivalent) in anhydrous DCM dropwise at 0° C. under a N atmosphere. The resulting mixture was stirred at 0° C. for 1 hour under a N atmosphere. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layer was separated, washed with brine, dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH) to give compound A4.

[0223] Example 2. Synthesis of 3-[(4-chloro-2-fluoro-5-methylphenyl)methyl]-1-cyclopropyl-1-[(3R)-1-(pyridazin-3-yl)piperidin-3-yl]urea [ka] Step 1: Synthesis of M2 To a mixture of M1 (10 g, 49.93 mmol) and K2CO3 (20.7 g, 149.8 mmol) in DMF (200 mL) was added 3,6-dichloropyridazine (7.44 g, 49.93 mmol). The resulting mixture was stirred at 80 °C for 16 h. After cooling, the resulting mixture was poured into water (800 mL). The precipitated solid was collected by filtration, washed with water (100 mL), and then dissolved in EtOAc. The organic layer was dried over Na2SO4, separated, and concentrated under reduced pressure to give M2 (12 g, 38.36 mmol, 76.83% yield) as a yellow solid. LC / MS (ESI) m / z: 313 (M+H) + .

[0224] Step 2: Synthesis of M3 To a solution of M2 (12 g, 38.36 mmol) in MeOH (200 mL) was added Pd / C (4.0 g, 10%). The resulting mixture was stirred under a hydrogen atmosphere at room temperature for 3 hours. The mixture was then filtered, and the filtrate was concentrated under reduced pressure to give M3 (10 g, 35.93 mmol, 93.63% yield) as a yellow solid. LC / MS (ESI) m / z: 279 (M+H) + .

[0225] Step 3: Synthesis of M4 To a solution of compound M3 (10 g, 35.93 mmol) in DCM (60 mL) was added TFA (60 mL). The resulting mixture was stirred at room temperature for 4 hours. The resulting mixture was concentrated to dryness under reduced pressure. The residue was diluted with MeOH, and NaHCO3 was added. The mixture was stirred at room temperature for 40 minutes and then the pH was adjusted to 8-9. The mixture was then filtered, and the filtrate was concentrated under reduced pressure to give M4 (6 g, 33.7 mmol, 93.7% yield) as a yellow oil. LC / MS (ESI) m / z: 179 (M+H) + .

[0226] Step 4: Synthesis of M5 To a mixture of compound M4 (5.8 g, 32.54 mmol) and 2,4-dimethoxybenzaldehyde (5.41 g, 32.54 mmol) in DCM (100 mL) was added AcOH (5.86 g, 97.62 mmol). The mixture was stirred at room temperature for 1 hour. Then, NaBH(OAc) (20.7 g, 97.62 mmol) was added to the above mixture. The resulting mixture was stirred for 3 hours. The mixture was then concentrated under reduced pressure. The residue was diluted with EtOAc and washed with aqueous NaHCO to pH = 8. The organic layer was washed with brine, dried over anhydrous NaSO, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with 5% MeOH / DCM) to give M5 (5 g, 13.7 mmol, 42.10% yield) as a yellow solid. LC / MS (ESI) m / z: 329 (M+H) + .

[0227] Step 5: Synthesis of M6 To a mixture of M5 (800 mg, 2.436 mmol) and (1-ethoxycyclopropoxy)trimethylsilane (1.06 g, 6.09 mmol) in EtOH (10 mL) and THF (20 mL) was added AcOH (2.19 g, 36.54 mmol) and NaBHCN (538 mg, 8.526 mmol). The resulting mixture was stirred at 80 °C for 16 h. After cooling, the resulting mixture was concentrated under reduced pressure. The residue was diluted with EtOAc and washed with water and brine. The organic layer was dried over anhydrous NaSO and concentrated to dryness. The crude product was purified by column chromatography on silica (eluted with 4% MeOH / DCM) to give M6 (560 mg, 1.52 mmol, 62.40% yield) as a yellow solid. LC / MS (ESI) m / z: 369 (M+H) + .

[0228] Synthesis of M7 Compound M6 (560 mg, 1.520 mmol) was added to TFA (18 mL), and the resulting mixture was stirred at 80° C. for 4 hours. After cooling, the mixture was concentrated under reduced pressure to give crude M7 (274 mg, 0.825 mmol, 54.26%) as a TFA salt. LC / MS (ESI) m / z: 219 (M+H) + .

[0229] 3-[(4-chloro-2-fluoro-5-methylphenyl)methyl]-1-cyclopropyl-1-[(3R)-1-(pyridazin-3-yl)piperidin-3-yl]urea To a mixture of M7 (274 mg, 0.824 mmol) in DCM (10 mL) was added TEA (251 mg, 2.473 mmol). After stirring at room temperature for 30 min, a solution of 2 (160 mg, 0.824 mmol) in DCM (2 mL) was added to the above mixture at 0° C. The resulting mixture was stirred at room temperature for 30 min. The mixture was then concentrated to dryness under reduced pressure. The crude product was purified by column chromatography on silica gel (eluted with 5% MeOH / DCM) to give Example 2 (103 mg, 0.247 mmol, 30.0% yield) as a pale yellow solid. LC / MS (ESI) m / z: 418 (M+H) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.50 (dd, J = 4.4, 1.0 Hz, 1H), 7.36 - 7.30 (m, 2H), 7.28 - 7.21 (m, 2H), 6.89 (t, J = 5.8 Hz, 1H), 4.41 - 4.25 (m, 4H), 3.68 - 3.60 (m, 1H), 3.23 - 3.16 (m, 1H), 2.81 - 2.67 (m, 1H), 2.50 - 2.44 (m, 1H), 2.29 (s, 3H), 2.15 - 2.07 (m, 1H), 1.91 - 1.70 (m, 2H), 1.54 - 1.42 (m, 1H), 0.93 - 0.86 (m, 2H), 0.74 - 0.62 (m, 2H). 19 F NMR (400 MHz, DMSO-d6) δ -121.05 (s).

[0230] The compounds in the following table were prepared from the appropriate starting materials described above or commercially available using General Procedure A above and Intermediate M7 from Example 2. [Table 6] TIFF0007797723000093.tif40169

[0231] Examples 7-127 The compounds in the following table were prepared from the appropriate starting materials, either as described above or commercially available, using the general procedure above and (R)-N-methyl-1-(pyridazin-3-yl)piperidin-3-amine (A3), or using intermediate M7 from Example 2.

[0232] [ka] tert-Butyl N-methyl-N-[(3R)-3-piperidyl]carbamate (2.14 g, 10 mmol), 3-bromopyridazine (2.38 g, 15.00 mmol), cesium fluoride (151.90 mg, 1.00 mmol, 36.87 uL), and potassium carbonate (3.46 g, 25.00 mmol, 1.51 mL) were dissolved in DMSO (100 mL) and heated to 150° C. overnight. The reaction mixture was then cooled to room temperature, poured into 600 mL of water, and extracted with ethyl acetate (3×200 mL). The combined organic layers were washed with water (1 × 200 mL) and brine (1 × 200 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude product, which was purified by flash column chromatography (ethyl acetate:heptane, 0:100 to 40:60) to give tert-butyl N-methyl-N-[(3R)-1-pyridazin-3-yl-3-piperidyl]carbamate A2 (1.57 g, 5.37 mmol, 53.70% yield). A2 (1.57 g, 5.37 mmol) was dissolved in DCM (4.2 mL) and TFA (6.12 g, 53.70 mmol, 4.14 mL) was added. The reaction mixture was stirred for 4 h. The reaction mixture was then concentrated under reduced pressure by rotary evaporation. The crude residue was dissolved in ethyl acetate, washed with 1M NaOH, brine, and concentrated to give (R)-N-methyl-1-(pyridazin-3-yl)piperidin-3-amine A3 (450 mg, 2.34 mmol, 43.59% yield), which was used without further purification. [Table 7] TIFF0007797723000096.tif250166 TIFF0007797723000097.tif245169 TIFF0007797723000098.tif250166 TIFF0007797723000099.tif250166 TIFF0007797723000100.tif250166 TIFF0007797723000101.tif250166 TIFF0007797723000102.tif250166 TIFF0007797723000103.tif250166 TIFF0007797723000104.tif250166 TIFF0007797723000105.tif240169 TIFF0007797723000106.tif234167 TIFF0007797723000107.tif249169 TIFF0007797723000108.tif250166 TIFF0007797723000109.tif240169 TIFF0007797723000110.tif250166 TIFF0007797723000111.tif246169 TIFF0007797723000112.tif250166 TIFF0007797723000113.tif250166 TIFF0007797723000114.tif250167 TIFF0007797723000115.tif250169 TIFF0007797723000116.tif248169 TIFF0007797723000117.tif250165 TIFF0007797723000118.tif240169 TIFF0007797723000119.tif250166 TIFF0007797723000120.tif250167 TIFF0007797723000121.tif250169 TIFF0007797723000122.tif139169

[0233] Example 128: Synthesis of 3-(4-chloro-3-methylbenzyl)-1-cyclopropyl-1-((R)-1-((S)-tetrahydrofuran-2-carbonyl)piperidin-3-yl)urea [ka] To a mixture of M2 (116 mg, 1.0 mmol) and HATU (474 ​​mg, 1.29 mmol) in DCM (20 mL) was added TEA (253 mg, 2.5 mmol) dropwise at 0 °C. After stirring at room temperature for 30 min, M1 (200 mg, 0.83 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for 2 h under a N2 atmosphere. The mixture was then diluted with DCM (20 mL) and washed with water (30 mL) and brine (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with PE / EtOAc = 100:0 to 3:1) to give M3 (250 mg, 88.8%) as a colorless oil. LC / MS (ESI) m / z: 299 (M+H) + To a solution of M3 (250 mg, 0.84 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise at 0° C. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was then evaporated to dryness under reduced pressure to give crude M4 (160 mg, 96.3%) as a yellow oil without further purification. LC / MS (ESI) m / z: 199 (M+H) +To a solution of M4 (160 mg, 0.81 mmol) and M5 (148 mg, 0.89 mmol) in DCM (10 mL) was added AcOH (145 mg, 2.42 mmol) at 0 °C. The resulting mixture was stirred at room temperature for 1 h. Then, NaBH(OAc) (510 mg, 2.42 mmol) was added dropwise to the above mixture at 0 °C. The resulting mixture was stirred overnight at room temperature under a N2 atmosphere. The mixture was quenched with saturated NaHCO3 solution (20 mL) and extracted twice with EtOAc (30 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was evaporated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with PE / EtOAc = 100:0 to = 2:1) to give M6 (100 mg, 35.6%) as a colorless oil. LC / MS (ESI) m / z: 349 (M+H) + To a mixture of M6 (100 mg, 0.29 mmol), M7 (125 mg, 0.72 mmol), and AcOH (52 mg, 0.86 mmol) in THF (12 mL) and EtOH (6 mL) was added NaBHCN (55 mg, 0.86 mmol). The resulting mixture was stirred overnight at 80 °C under a N2 atmosphere. After cooling, the reaction was quenched with saturated aqueous NaHCO3 (20 mL) and extracted twice with EtOAc (30 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was evaporated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluted with PE / EtOAc = 100:0 to = 2:1) to give M8 (50 mg, 44.8%) as a colorless oil. LC / MS (ESI) m / z: 389 (M+H) + A solution of M8 (50 mg, 0.13 mmol) in TFA (4 mL) was stirred at 80 °C under N2 atmosphere for 3 h. After cooling, the mixture was concentrated to dryness under reduced pressure to give crude M9 (30 mg, 97.8%) as a purple oil without further purification. LC / MS (ESI) m / z: 239 (M+H) + .

[0234] To a mixture of M9 (30 mg, 0.13 mmol) and TEA (39 mg, 0.39 mmol) in anhydrous DCM (10 mL) was added dropwise a solution of M10 (22.9 mg, 0.13 mmol) in anhydrous DCM (2 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 1 h under a N atmosphere. The reaction mixture was then diluted with water (20 mL) and extracted twice with DCM (20 mL). The combined organic layer was separated, dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with DCM / MeOH = 100:0 to 20:1) to give the crude product. The crude product was purified by preparative HPLC (Gemini 5 μm C18 250*21.2 mm, HO / MeCN (5-95%) / 0.1% HCOOH) to give Example 128 (11 mg, 0.026 mmol, 20.81% yield) as a white solid. LC / MS (ESI) m / z: 420 (M+H) + .1H NMR (400 MHz, MeOD) δ 7.28 (dd, J = 8.2, 2.7 Hz, 1H), 7.21 (d, J = 5.1 Hz, 1H), 7.10 (dd, J = 10.5, 4.1 Hz, 1H), 4.74 - 4.67 (m, 1H), 4.53 - 4.39 (m, 1H), 4.39 - 4.26 (m, 2H), 4.02 - 3.87 (m, 2H), 3.86 - 3.44 (m, 2H), 3.30 - 3.10 (m, 1H), 3.00 - 2.46 (m, 2H), 2.33 (d, J = 9.2 Hz, 3H), 2.32 - 2.13 (m, 2H), 2.05 - 1.79 (m, 5H), 1.66 - 1.39 (m, 1H), 1.02 - 0.87 (m, 2H), 0.85 - 0.66 (m, 2H).

[0235] The compounds in the following table were made by the same route as Example 128, starting from the appropriate commercially available amide M2 ​​and other intermediates described above or available commercially. [Table 8]

[0236] The following examples were synthesized according to general procedure A using commercially available building blocks. [Table 9] TIFF0007797723000126.tif250167 TIFF0007797723000127.tif233169 TIFF0007797723000128.tif167167

[0237] Synthesis of General Intermediates, Methods B-G [ka]

[0238] General Procedure B: [ka] To a mixture of compound B1 (1.0 equivalent) and aryl halide (1.1 equivalent) in DMSO (0.1 mol / L) was added K2CO3 (3.0 equivalents) and CsF (0.2 equivalents). The resulting mixture was stirred at 90 °C for 16 hours. The mixture was then diluted with HO and extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by column chromatography on silica gel to give compound B2.

[0239] General procedure B synthesis example: [ka] To a mixture of compound B3 (100 mg, 0.47 mmol) and compound 2 (101 mg, 0.47 mmol) in DMSO (8 mL) was added K2CO3 (194 mg, 1.40 mmol) and CsF (15 mg, 0.09 mmol). The resulting mixture was stirred at 90 °C for 16 h. The mixture was then diluted with HO (30 mL) and extracted with EtOAc (20 mL * 2). The combined organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with PE: EtOAc = 10:1 to 2:1) to give compound B4 (140 mg, 87.5% yield) as a yellow solid. LC / MS (ESI) m / z: 349 (M+H) + .

[0240] General Procedure C: [ka] To a mixture of compound C1 (1.2 equivalents) and aryl halide (1.0 equivalents) in toluene was added t-BuONa (2.0 equivalents), DavePhos (0.1 equivalents), and Pd(OAc) (0.1 equivalents) under a N atmosphere. The resulting mixture was stirred at 100° C. for 16 hours under a N atmosphere. The mixture was then diluted with EtOAc, filtered, and concentrated to dryness. The residue was purified by column chromatography on silica gel to give compound C2.

[0241] General procedure C synthesis example: [ka] To a mixture of C3 (0.58 g, 2.72 mmol) and C4 (0.3 g, 2.09 mmol) in toluene (15 mL) was added t-BuONa (410 mg, 4.18 mmol), DavePhos (83 mg, 0.21 mmol), and Pd(OAc) (47 mg, 0.21 mmol) under a N atmosphere. The resulting mixture was stirred at 120 °C for 16 h under a N atmosphere. The mixture was then diluted with EtOAc (30 mL), filtered, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with PE:EtOAc = 50:1 to 20:1) to give compound C5 (0.58 g, 87% yield) as a colorless oil. LC / MS (ESI) m / z: 322 (M+H) + .

[0242] General Procedure D: [ka] To a mixture of amine D1 (1.1 equiv.) and aryl halide (1.0 equiv.) was added a solution of LiHMDS (5.0 equiv., 1 M), RuPhos (0.1 equiv.), and Pd(OAc) (0.1 equiv.) in THF under N2 atmosphere. The resulting mixture was stirred at 80 °C under N2 atmosphere for 4 h. The mixture was then quenched with aqueous NH4Cl and extracted twice with EtOAc. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel to give compound D2.

[0243] General procedure D synthesis example: [ka] A mixture of D2 (153 mg, 0.72 mmol) and D3 (100 mg, 0.65 mmol) in THF (12 mL) was treated with LiHMDS (3.3 mL, 1 M in THF), RuPhos (31 mg, 0.07 mmol) and Pd(OAc )2(15 mg, 0.07 mmol) was added under N2 atmosphere. The resulting mixture was stirred at 80 °C under N2 atmosphere for 4 h. The mixture was then quenched with aqueous NH4Cl (30 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with PE: EtOAc = 50:1 to 5:1) to give D4 (115 mg, 53% yield) as a pale yellow solid. LC / MS (ESI) m / z: 332 (M+H) + .

[0244] General Step E example: [ka] To a mixture of E1 (100 mg, 0.65 mmol) and E2 (182 mg, 0.85 mmol) in THF (18 mL) was added XPhos-Pd-G1 (25 mg, 0.03 mmol), XPhos (33 mg, 0.07 mmol), and LiHMDS (2.6 mL, 1 M in THF). The resulting mixture was stirred at 80 °C for 16 h under a N2 atmosphere. The mixture was then quenched with aqueous NH4Cl (40 mL) and extracted twice with EtOAc (25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with PE:EtOAc = 20:1 to 3:1) to give E3 (130 mg, 60% yield) as a pale yellow oil. LC / MS (ESI) m / z: 332 (M+H) + .

[0245] General Step F Example: [ka] To a mixture of F1 (200 mg, 1.311 mmol) and F2 (280 mg, 1.311 mmol) in THF (20 mL) was added RuPhos (61 mg, 0.131 mmol) and RuPhos-Pd-G2 (101 mg, 0.131 mmol). After stirring at 0 °C for 10 min, LiHMDS (3.14 mL, 1 M in THF) was added dropwise, and the resulting mixture was stirred at 70 °C for an additional 3 h under a N2 atmosphere. After cooling, the mixture was quenched with aqueous NH4Cl (30 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel (eluting with PE:EtOAc = 50:1 to 2:1) to give F3 (160 mg, 37% yield) as a yellow oil. LC / MS (ESI) m / z: 331(M+H) + .

[0246] General Procedure G example: [ka] To a solution of G2 (200 mg, 1.235 mmol) and G1 (265 mg, 1.235 mmol) in DMSO (10 mL) was added CuI (47 mg, 0.247 mmol), L-proline (28 mg, 0.247 mmol), and K2CO3 (512 mg, 3.704 mmol), and the resulting mixture was stirred at 100 °C under a N2 atmosphere for 16 h. After cooling, the mixture was quenched with aqueous NH4Cl (30 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:1 to 20:1) to give G3 (160 mg, 44% yield) as a yellow oil. LC / MS (ESI) m / z: 296 (M+H) + .

[0247] The examples in the table below were prepared using methods analogous to those described above from the appropriate starting materials or commercially available starting materials. [Table 10] TIFF0007797723000140.tif250162 TIFF0007797723000141.tif250160 TIFF0007797723000142.tif250160 TIFF0007797723000143.tif250165 TIFF0007797723000144.tif250158 TIFF0007797723000145.tif252168 TIFF0007797723000146.tif250160 TIFF0007797723000147.tif236168 TIFF0007797723000148.tif250165 TIFF0007797723000149.tif145168

[0248] General Procedure H: As a general procedure, the following examples were synthesized according to the following general scheme. [ka] To a solution of compound H1 (1 equivalent) in toluene, TEA (2 equivalents) and DPPA (1.2 equivalents) were added. The resulting mixture was stirred at 110°C under a N2 atmosphere for 2 hours. After cooling, the resulting mixture was concentrated to dryness under reduced pressure. The crude compound was used directly in the next step without further purification. To a solution of compound H3 (1 equivalent) in DCM, TEA (3 equivalents) and compound H2 (1 equivalent) were added at 0°C. The resulting mixture was stirred at room temperature under a N2 atmosphere for 30 minutes. The mixture was then concentrated to dryness under reduced pressure. The crude product was purified by column chromatography on silica gel (eluted with DCM / MeOH) to give compound H4.

[0249] Example 184: Synthesis of 3-((1S,2R)-2-(4-chloro-3-methylphenyl)cyclopropyl)-1-cyclopropyl-1-((R)-1-(pyridazin-3-yl)piperidin-3-yl)urea [ka] To a solution of M1 (200 mg, 0.952 mmol) in toluene (10 mL), TEA (192 mg, 1.904 mmol) and DPPA (314 mg, 1.142 mmol) were added. The resulting mixture was stirred at 110 °C under a N2 atmosphere for 2 h. After cooling, the mixture was concentrated under reduced pressure to give crude M2 ​​(197 mg, 99.9% yield) without further purification. To a solution of M3 (207 mg, 0.952 mmol) in anhydrous DCM (10 mL), TEA (288 mg, 2.856 mmol) and M2 (197 mg, 0.952 mmol) were added at 0 °C. The resulting mixture was stirred at room temperature under a N2 atmosphere for 30 min. The mixture was then concentrated to dryness under reduced pressure. The crude product was purified by column chromatography on silica gel (eluting with 1% MeOH / DCM to 5% MeOH / DCM) to give Example 184 (61.1 mg, 15.1% yield) as a white solid. LC-MS: m / z 426 (M+H) +.1H NMR (400 MHz, MeOD) δ 8.43 (dd, J = 4.4, 1.2 Hz, 1H), 7.37 (dd, J = 9.4, 4.4 Hz, 1H), 7.29 (dd, J = 9.4, 1.2 Hz, 1H), 7.21 (d, J = 8.2 Hz, 1H), 7.09 (d, J = 1.9 Hz, 1H), 6.95 (dd, J = 8.2, 2.1 Hz, 1H), 6.69 (s, 1H), 4.43 - 4.31 (m, 2H), 3.80 - 3.69 (m, 1H), 2.91 - 2.81 (m, 1H), 2.78 - 2.71 (m, 1H), 2.51 - 2.45 (m, 1H), 2.32 (s, 3H), 2.30 - 2.20 (m, 1H), 2.03 - 1.94 (m, 2H), 1.90 - 1.83 (m, 1H), 1.66 - 1.57 (m, 1H), 1.24 - 1.14 (m, 2H), 0.95 - 0.89 (m, 2H), 0.79 - 0.72 (m, 2H).

[0250] The compounds in the following table were prepared using General Procedure B above, detailed in Example 184, from the appropriate starting materials described above or commercially available. [Table 11] TIFF0007797723000153.tif250167 TIFF0007797723000154.tif250167 TIFF0007797723000155.tif238169 TIFF0007797723000156.tif238169 TIFF0007797723000157.tif243169 TIFF0007797723000158.tif250167 TIFF0007797723000159.tif250167 TIFF0007797723000160.tif243169 TIFF0007797723000161.tif250167 TIFF0007797723000162.tif250167 TIFF0007797723000163.tif233169 TIFF0007797723000164.tif250167 TIFF0007797723000165.tif250167 TIFF0007797723000166.tif46167

[0251] General Procedure I: As a general procedure, the following examples were synthesized according to the following general scheme. [ka] To a mixture of compound I1 (1 equivalent) and DIEA (5 equivalents) in DMF was added the corresponding acid I2 (1.2 equivalents), EDCI (1.2 equivalents), and HOBt (1.2 equivalents). The resulting mixture was stirred at room temperature for 16 hours. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness in vacuo. The residue was purified by preparative HPLC (Xbudge prep C18 250*19mm 5um OBD, HO / MeCN (5-95%) / 0.1% FA) to give compound I3.

[0252] Example 232: Synthesis of (R)-3-(4-chloro-3-methylbenzyl)-1-methyl-1-(1-(pyrimidine-4-carbonyl)piperidin-3-yl)urea [ka] To a mixture of compound M1 (50 mg, 0.12 mmol) and DIEA (79 mg, 0.61 mmol) in DMF was added pyrimidine-4-carboxylic acid M2 (19 mg, 0.14 mmol), EDCI (28 mg, 0.144 mmol), and HOBt (20 mg, 0.144 mmol). The resulting mixture was stirred at room temperature for 16 hours. The mixture was then diluted with water (20 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness under vacuum. The residue was purified by preparative HPLC (Xbudge prep C18 250*19 mm 5 um OBD, HO / MeCN (5-95%) / 0.1% FA) to give Example 232 (9 mg, 18.7% yield) as a white solid. LC / MS: m / z 402 (M+H) + .1H NMR (400 MHz, MeOD) δ 9.21 - 9.12 (m, 1H), 8.94 - 8.90 (m, 1H), 7.66 - 7.63 (m, 1H), 7.28 - 7.23 (m, 1H), 7.14 (s, 1H), 7.12 - 7.00 (m, 1H), 4.64 - 4.49 (m, 1H), 4.38 - 4.11 (m, 3H), 3.70 - 3.54 (m, 1H), 3.28 - 3.18 (m, 1H), 3.08 - 2.42 (m, 5H), 2.36 - 2.29 (m, 3H), 1.98 - 1.76 (m, 3H), 1.76 - 1.62 (m, 1H).

[0253] The compounds in the following table were prepared from the appropriate starting materials described above or commercially available using General Procedure I above and Intermediate M1 of Example 232. [Table 12] TIFF0007797723000170.tif237169 TIFF0007797723000171.tif243169 TIFF0007797723000172.tif245167 TIFF0007797723000173.tif233167 TIFF0007797723000174.tif224167 TIFF0007797723000175.tif234169 TIFF0007797723000176.tif240167 TIFF0007797723000177.tif240167 TIFF0007797723000178.tif181169

[0254] General Procedure J: Example 272: Synthesis of (R)-3-(4-chloro-3-methylbenzyl)-1-(1-(cyclopropanecarbonyl)piperidin-3-yl)-1-cyclopropylurea [ka] To a mixture of J1 (67 mg, 0.21 mmol) and DIEA (81 mg, 0.63 mmol) in anhydrous DMF (5 mL) was added cyclopropanecarboxylic acid J2 (19 mg, 0.22 mmol) and HATU (84 mg, 0.22 mmol). The resulting mixture was stirred at room temperature for 1 h. The mixture was then poured into water (10 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give Example 272 (17 mg, 20.73% yield) as a pale yellow solid. LC / MS (ESI) m / z: 390 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.27 (d, J = 8.2 Hz, 1H), 7.24 - 7.17 (m, 1H), 7.13 - 7.05 (m, 1H), 4.54 - 4.39 (m, 1H), 4.37 - 4.21 (m, 3H), 3.57 - 3.34 (m, 1H), 3.22 - 2.91 (m, 1H), 2.65 - 2.43 (m, 2H), 2.34 (s, 3H), 2.28 - 2.12 (m, 1H), 1.99 - 1.76 (m, 3H), 1.62 - 1.36 (m, 1H), 0.97 - 0.68 (m, 8H).

[0255] The compounds in the following table were prepared from the appropriate starting materials described above or commercially available using General Procedure J above and Intermediate J1 of Example 272. [Table 13]

[0256] General steps: Example 274: Synthesis of (R)-3-(4-chloro-3-methylbenzyl)-1-methyl-1-(1-(6-(trifluoromethyl)pyridazin-3-yl)piperidin-3-yl)urea [ka] To a mixture of K1 (59 mg, 0.2 mmol) and K2CO3 (56 mg, 0.4 mmol) in DMF (6 mL) was added 3-chloro-6-(trifluoromethyl)pyridazine K2 (44 mg, 0.24 mmol). The resulting mixture was stirred at 80 °C for 10 h. After cooling, the mixture was diluted with saturated NH4Cl solution (20 mL) and extracted twice with EtOAc (20 mL). The combined organic layers were washed with saturated NH4Cl solution and brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give Example 274 (8 mg, 9.1% yield) as a white solid. LC / MS (ESI) m / z: 442 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.64 (d, J = 9.7 Hz, 1H), 7.35 (d, J = 9.7 Hz, 1H), 7.27 (d, J = 8.2 Hz, 1H), 7.24 - 7.20 (m, 1H), 7.11 (dd, J = 8.2, 1.7 Hz, 1H), 4.57 - 4.47 (m, 1H), 4.45 - 4.36 (m, 1H), 4.31 (s, 2H), 4.12 - 4.02 (m, 1H), 3.19 - 3.09 (m, 1H), 3.04 - 2.94 (m, 1H), 2.90 (s, 3H), 2.33 (s, 3H), 1.98 - 1.84 (m, 3H), 1.75 - 1.60 (m, 1H). [Table 14]

[0257] The following examples were prepared according to general procedure A. [Table 15] TIFF0007797723000184.tif234169 TIFF0007797723000185.tif250162 TIFF0007797723000186.tif192167

[0258] General Procedure: [ka] Step 1 To a solution of (9H-fluoren-9-yl)methyl (R)-3-(cyclopropylamino)piperidine-1-carboxylate (1 equivalent) and TEA (3 equivalents) in anhydrous DCM, a solution of compound M1 (1 equivalent) in anhydrous DCM was added dropwise at 0 ° C. under a N atmosphere. The resulting mixture was stirred at 0 ° C. under a N atmosphere for 1 hour. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layer was separated, washed with brine, dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with heptane / ethyl acetate) to give compound M2.

[0259] Step 2 M2 (1 equiv.) was dissolved in 20% piperidine / DMF (0.1 M) and the reaction mixture was stirred at room temperature for 1-2 h. The reaction mixture was then concentrated in vacuo and the crude product was purified by flash column chromatography using DCM / DCM:7N NH4 MeOH (10:0-0:10) to give product M3.

[0260] Step 3 M3 (1 eq.), the appropriate carboxylic acid M4 (1.2 eq.), and HATU (1.3 eq.) were dissolved in DMF (0.1 M), then DIEA (2 eq.) was added, and the reaction was stirred at room temperature until complete conversion. Water was added, and the reaction mixture was extracted twice with DCM. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give the pure product M5.

[0261] Synthesis of 1-cyclopropyl-1-[(3R)-1-(2-hydroxyacetyl)piperidin-3-yl]-3-[(3-phenyl-1,2-oxazol-5-yl)methyl]urea, Example 290 [ka] To a solution of (9H-fluoren-9-yl)methyl (R)-3-(cyclopropylamino)piperidine-1-carboxylate (400 mg, 1.10 mmol) and TEA (0.460 mL, 3.30 mmol) in anhydrous DCM (11 mL) was added a solution of 5-(isocyanatomethyl)-3-phenylisoxazole (220 mg, 1.10 mmol) in anhydrous DCM (5.5 mL) dropwise at 0 °C under a N atmosphere. The resulting mixture was stirred at 0 °C for 1 h under a N atmosphere. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layer was separated, washed with brine, dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with heptane / ethyl acetate) to give X1 (280 mg, 45% yield). X1 (280 mg, 0.498 mmol) was dissolved in 20% piperidine / DMF (4.98 mL), and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was then concentrated in vacuo, and the crude product was purified by flash column chromatography using DCM / DCM:7N NH4 MeOH (10 / 0 to 0 / 10) to give X2 (150 mg, 88% yield). X2 (20 mg, 0.06 mmol), 2-hydroxyacetic acid (5.4 mg, 0.072 mmol), and HATU (29 mg, 0.076 mmol) were dissolved in DMF (0.56 mL), and then DIEA (21 μL, 0.12 mmol) was added, and the reaction was stirred at room temperature for 5 min. Water was added, and the reaction mixture was extracted twice with DCM. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give Example 290 (11 mg, 47% yield) as a white powder. LC / MS (ESI) m / z: 399.1 (M+H) + .

[0262] The following examples were prepared according to general procedure M. [Table 16] TIFF0007797723000190.tif224169 TIFF0007797723000191.tif250163 TIFF0007797723000192.tif251169 TIFF0007797723000193.tif250161 TIFF0007797723000194.tif219167 TIFF0007797723000195.tif224169 TIFF0007797723000196.tif250163 TIFF0007797723000197.tif36169

[0263] General Procedure N [ka] To a solution of compound (R)-N-cyclopropyl-1-((2-nitrophenyl)sulfonyl)piperidin-3-amine (1 equivalent) and TEA (3 equivalents) in anhydrous DCM was added dropwise a solution of compound N1 (1 equivalent) in anhydrous DCM under a N2 atmosphere at 0°C. The resulting mixture was stirred at 0°C for 1 hour under a N2 atmosphere. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with heptane / ethyl acetate) to give compound N2. N2 (1 equivalent) was dissolved in DMF (0.1 M), potassium carbonate (2 equivalents) and thiophenol (1.5 equivalents) were added, and the reaction mixture was stirred at room temperature for 1-2 hours. 1 M HCl was added until the pH reached 2-3, and the resulting mixture was extracted twice with DCM. The acidic solution was then brought to pH 10-11 by careful addition of NaOH 6N. The resulting basic solution was then extracted with DCM (x3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude N3 was used in the next step without further purification. N3 (1 equiv.), the appropriate carboxylic acid N4 (1.2 equiv.), and HATU (1.3 equiv.) were dissolved in DMF (0.1 M), then DIEA (2 equiv.) was added, and the reaction was stirred at room temperature until complete conversion. Water was added, and the reaction mixture was extracted twice with DCM. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The crude product was purified by preparative HPLC to give the pure product N5.

[0264] General steps: [ka] O1 was synthesized using procedures M or N.

[0265] Synthesis of O3 The appropriate amine O2 (2 equiv.) and CDI (2 equiv.) were dissolved in DMF (0.1 M) and the reaction was stirred at room temperature for 2-3 h. O1 (1 equiv.) was then added and the reaction was stirred at 50 °C until complete conversion (1-4 h). The reaction mixture was concentrated in vacuo and the crude was purified by preparative HPLC to give the product O3.

[0266] Synthesis of (3R)-N-cyclopropyl-3-(3-cyclopropyl{[(4-chloro-2-fluoro-5-methylphenyl)methyl]carbamoyl}amino)piperidine-1-carboxamide (Example 317) [ka] Cyclopropylamine (8.3 uL, 0.12 mmol) and CDI (19.5 mg, 0.12 mmol) were dissolved in DMF, and the reaction was stirred at room temperature for 2 h. X1 (20 mg, 0.06 mmol) was then added, warmed to 50 °C, and stirred for 2 h. The reaction mixture was then cooled to room temperature and concentrated in vacuo. The crude material was purified by preparative HPLC to give X1 (10 mg, 39% yield) as a white powder. LC / MS (ESI) m / z: 423.1 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.24 (d, J = 8.0 Hz, 1H), 7.13 (d, J = 9.8 Hz, 1H), 6.86 (t, J = 5.7 Hz, 1H), 4.37 (t, J = 4.7 Hz, 2H), 3.96 (d, J = 12.8 Hz, 1H), 3.84 (d, J = 12.5 Hz, 1H), 3.61 (ddd, J = 11.8, 7.9, 4.1 Hz, 1H), 3.13 - 3.06 (m, 1H), 2.61 (td, J = 13.0, 2.4 Hz, 1H), 2.55 - 2.47 (m, 2H), 2.32 (s, 3H), 2.07 (td, J = 12.5, 3.8 Hz, 1H), 1.87 (d, J = 11.6 Hz, 1H), 1.74 (d, J = 13.0 Hz, 1H), 1.47 (dt, J = 13.3, 4.2 Hz, 1H), 0.96 - 0.89 (m, 2H), 0.73 (d, J = 3.6 Hz, 2H), 0.66 - 0.60 (m, 2H), 0.47 - 0.40 (m, 2H). [Table 17] TIFF0007797723000202.tif97169

[0267] General steps: [ka] The protected intermediate 3-amino-piperidine can be prepared by the general route or general procedure Q above.

[0268] A solution of tert-butyl N-[(3R)-3-piperidyl]carbamate P1 (25 g, 124.83 mmol) and DIPEA (19.36 g, 149.79 mmol, 26.09 mL) in 500 mL of DCM was added dropwise to a solution of Fmoc-Cl (44.28 g, 137.31 mmol) in DCM (150 mL) over 2 h in an ice bath. The reaction mixture was then stirred for an additional 1 h, then warmed to room temperature, filtered, and concentrated to give 9H-fluoren-9-ylmethyl (3R)-3-(tert-butoxycarbonylamino)piperidine-1-carboxylate P2 (81 g, 191.71 mmol, 153.58% yield) as a crude white solid, which was used without further purification. To a suspension of 9H-fluoren-9-ylmethyl (3R)-3-(tert-butoxycarbonylamino)piperidine-1-carboxylate P2 (20 g, 47.34 mmol) in THF (200 mL) was added 4.0 M dioxane / HCl (4.0 M, 59.17 mL), and the reaction mixture was stirred for 3 h at 60 °C. The reaction mixture was then concentrated under reduced pressure by rotary evaporation, and the resulting solid was washed twice with EtO to give 9H-fluoren-9-ylmethyl (3R)-3-aminopiperidine-1-carboxylate P3 (13.91 g, 38.76 mmol, 81.89% yield, HCl) as a white solid. To a suspension of 9H-fluoren-9-ylmethyl (3R)-3-aminopiperidine-1-carboxylate P3 (13.91 g, 38.76 mmol, HCl) in DCM (250 mL) was added 2,4-dimethoxybenzaldehyde (6.12 g, 36.82 mmol), DIPEA (5.51 g, 42.64 mmol, 7.43 mL), and sodium triacetoxyborohydride (8.22 g, 38.76 mmol). The reaction mixture was stirred overnight, then washed with 1 M NaOH, brine, and dried over anhydrous sodium sulfate. The reaction mixture was then concentrated and purified by chromatography (DCM:MeOH 100:0 to 95:5) to give 9H-fluoren-9-ylmethyl (3R)-3-[(2,4-dimethoxyphenyl)methylamino]piperidine-1-carboxylate P4 (9.26 g, 19.59 mmol, 50.55% yield) as a yellow oil.To a solution of 9H-fluoren-9-ylmethyl (3R)-3-[(2,4-dimethoxyphenyl)methylamino]piperidine-1-carboxylate (5.75 g, 12.17 mmol) P4 in THF (50 mL) and ethanol (100 mL), (1-ethoxycyclopropoxy)-trimethyl-silane (5.30 g, 30.43 mmol, 6.12 mL), acetic acid (10.96 g, 182.55 mmol, 10.44 mL), and sodium cyanoborohydride (2.68 g, 42.60 mmol) were added. The reaction mixture was stirred at 80 °C overnight, then concentrated, and the residue was dissolved in DCM. The organic layer was washed with saturated aqueous sodium carbonate, brine, and dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by chromatography (DCM:MeOH 10:0 to 9:1) to give 9H-fluoren-9-ylmethyl (3R)-3-[cyclopropyl-[(2,4-dimethoxyphenyl)methyl]amino]piperidine-1-carboxylate P5 as a white foam. A solution of 9H-fluoren-9-ylmethyl (3R)-3-[cyclopropyl-[(2,4-dimethoxyphenyl)methyl]amino]piperidine-1-carboxylate P5 (2.0 g, 3.90 mmol) in TFA (59.20 g, 519.19 mmol, 40 mL) was heated to 80 °C and stirred for 6 h. The reaction mixture was then cooled to room temperature and concentrated. The residue was dissolved in DCM, washed with saturated aqueous Na2CO3, dried over anhydrous Na2SO4, and concentrated to give 9H-fluoren-9-ylmethyl (3R)-3-(cyclopropylamino)piperidine-1-carboxylate P6 (1.35 g, 3.72 mmol, 95.47% yield) as a brown oil, which was stored at -20 °C and used without further purification.

[0269] General Procedure Q: [ka] To a solution of tert-butyl N-[(3R)-3-piperidyl]carbamate (10 g, 49.93 mmol) Q1 and DIPEA (7.10 g, 54.92 mmol, 9.57 mL) in DCM (500 mL) was added 2-nitrobenzenesulfonyl chloride (11.62 g, 52.43 mmol) as a solid in portions. The reaction mixture was stirred at ambient temperature for 15 minutes, and then TFA (56.93 g, 499.31 mmol, 38.47 mL) was added slowly. The reaction mixture was stirred at ambient temperature for an additional 2 hours and then concentrated under reduced pressure by rotary evaporation. The crude residue was then resuspended in 500 mL of DCM, and to this solution was added DIPEA (11.29 g, 87.38 mmol, 15.22 mL), 2,4-dimethoxybenzaldehyde (7.88 g, 47.43 mmol), and sodium triacetoxyborohydride (26.46 g, 124.83 mmol). The reaction mixture was stirred overnight at ambient temperature. The reaction mixture was then washed with 1 M NaOH (500 mL), and the organic layer was then separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure by rotary evaporation. The crude residue was purified by flash column chromatography (DCM:MeOH, 100:0 to 96:4) to afford (3R)-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (7.94 g, 18.23 mmol, 36.52% yield) Q2. To a solution of (3R)-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (7.93 g, 18.21 mmol) Q2 in THF (240 mL) and EtOH (120 mL) was added 1-ethoxy-1-trimethylsiloxycyclopropane (7.94 g, 45.52 mmol, 9.15 mL), sodium cyanoborohydride (4.01 g, 63.73 mmol), and acetic acid (16.40 g, 273.14 mmol, 15.62 mL). The reaction mixture was stirred at 80° C. overnight, then cooled to room temperature and concentrated under reduced pressure by rotary evaporation.The residue was then dissolved in ethyl acetate (250 mL), washed with 1 M NaOH (250 mL), brine (250 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure by rotary evaporation to give (3R)-N-cyclopropyl-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (8.58 g, 18.04 mmol, 99.08% yield) Q3, which was used without further purification. (3R)-N-cyclopropyl-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (8.58 g, 18.04 mmol) was dissolved in TFA (100 mL) Q3 and Et3SiH (10 mL) and heated to 80 °C. After 4 h, an additional portion of triethylsilane (7.28 g, 62.61 mmol, 10 mL) was added to quench the formation of the dimethoxytolyl cation. The reaction mixture was then stirred overnight. The reaction mixture was then concentrated under reduced pressure by rotary evaporation, and the crude residue was dissolved in 200 mL of ethyl acetate. The organic layer was washed with 3 M NaOH (approximately 200 mL), brine (approximately 200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure by rotary evaporation. The crude residue was then dissolved in 100 mL of diethyl ether, and a 2.0 M solution of hydrogen chloride in diethyl ether (2.0 M, 9.02 mL) was added slowly dropwise. The product was filtered from the solution to give (3R)-N-cyclopropyl-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (5.5 g, 15.20 mmol, 84.25% yield, HCl) Q4 as a tan solid.

[0270] General Procedure The benzylamine intermediate attached to the heterocycle on the aromatic ring was prepared by the general procedure. [ka] The appropriate aryl bromide R1 (1 equiv.) was dissolved in dioxane (0.3 M), then the corresponding dilute reagent R2 (1.1 equiv.) and palladium(0) tetrakis(triphenylphosphine) (0.1 equiv.) were added, and the reaction was stirred at 110 °C overnight. The reaction mixture was then cooled and concentrated in vacuo. The crude was purified by flash column chromatography using heptane / ethyl acetate to give the pure product R3. R3 (1 equiv.) was dissolved in HCl 4 M dioxane (20 equiv.), and the reaction was stirred at room temperature until complete deprotection. Saturated NaHCO3 solution was added, and the reaction mixture was extracted with DCM (x3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product R4 was used in the next step without further purification.

[0271] Synthesis of (2-fluoro-4-(oxazol-2-yl)phenyl)methanamine R6 by general procedure R [ka] R4 (520 mg, 1.71 mmol) was dissolved in dioxane (5.12 mL), then 2-(tributylstannyl)oxazole (673 mg, 1.88 mmol) and palladium(0) tetrakis(triphenylphosphine) (198 mg, 0.17 mmol) were added, and the reaction was stirred at 110 °C overnight. The reaction mixture was then cooled and concentrated in vacuo. The crude material was purified by flash column chromatography using heptane / ethyl acetate to give the pure product R5 (350 mg, 70% yield) as a white powder. R5 (350 mg, 1.20 mmol) was dissolved in HCl 4 M dioxane (6 mL, 24 mmol), and the reaction was stirred at room temperature until complete deprotection. Saturated NaHCO3 solution was added, and the reaction mixture was extracted with DCM (x3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product (2-fluoro-4-(oxazol-2-yl)phenyl)methanamine R6 was used in the next step without further purification.

[0272] Examples 323-333 were prepared from the corresponding aryl- or heteroaryl-benzylamines and the general procedures described above. [Table 18] TIFF0007797723000208.tif250165 TIFF0007797723000209.tif250162

[0273] General Procedure R' described below was used to prepare Examples 334-339 and 343 and similar compounds. [ka]

[0274] General Procedure S described below was used to prepare Examples 340, 341, and 342 and 343 and similar compounds. [ka]

[0275] Examples 334 to 343 [Table 19] TIFF0007797723000213.tif250157 TIFF0007797723000214.tif250157 TIFF0007797723000215.tif143165

[0276] General procedure R was used to prepare the final acid, amide or sulfonamide in the final step according to the previous general procedure in the following examples.

[0277] Example 160 was prepared by general procedure C followed by general procedure R.

[0278] Examples 166, 192, 193, 201, 206, 212 were prepared by general procedure B followed by general procedure R.

[0279] Example 335 was prepared by general procedure C followed by general procedure R.

[0280] Example 200 was prepared by general procedure D followed by general procedure R.

[0281] General Procedure S was used to prepare the final amine or amide in the final step according to the previous general procedure in the following examples.

[0282] Example 180 was prepared by general procedure C followed by general procedure S.

[0283] Representative compounds selected from above were tested in the isoleucine transport assay described in Example 1. The results are summarized in the table in FIG.

[0284] General Procedure S' [ka] Step 1: Synthesis of S2 To a mixture of compound S1 (1 equivalent) and TEA (4 equivalents) in anhydrous THF (0.05 M) was added TMSNCO (1.5 equivalents) dropwise under a N atmosphere at 0 °C, and the resulting mixture was stirred at room temperature for 16 hours. The mixture was then poured into H O and washed twice with EtOAc. The combined organic layer was dried over anhydrous Na SO , filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM:MeOH) to give compound S2.

[0285] Step 2: Composition of S3 S2 (1 equiv.) was dissolved in a solution of DCM / TFA 5:1 (0.1 M), and the resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give crude compound S3, which was used in the next step without further purification.

[0286] Step 3: Synthesis of S4 To a solution of S3 (1 eq.) in anhydrous DCM (0.03 M), 2,4-dimethoxybenzaldehyde (1.2 eq.) and acetic acid (2 eq.) were added, and the resulting mixture was stirred at room temperature for 1 hour. Then, NaBH(OAc)3 (2 eq.) was added to the above mixture, and the resulting mixture was stirred at 45 °C overnight. The mixture was then poured into 5% Na2CO3 solution and extracted with DCM (x4). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH) to give compound S4.

[0287] Step 4: Composition of S5 To a mixture of S4 (1 equivalent) and acetic acid (10 equivalents) in THF / EtOH 1:2 (0.03 M) was added (1-ethoxycyclopropoxy)trimethylsilane (1.1 equivalents), followed by NaBHCN (3 equivalents), and the resulting mixture was stirred at 80 °C for 16 h under a N atmosphere. The mixture was then poured into 5% NaCO solution and extracted with DCM (x3). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with DCM / MeOH) to give compound S5.

[0288] Step 5: Compositing S6 S5 (1 equivalent) was added in TFA (0.1 M), and the resulting mixture was stirred at 80° C. for 16 hours. The mixture was then concentrated to dryness under reduced pressure, and the crude product was purified by flash column chromatography (eluted with DCM / MeOH) to give compound S6.

[0289] Step 6: Composition of S8 To a solution of S6 (1 equiv.) in anhydrous DCM (0.03 M) was added DIEA (3 equiv.), and the resulting mixture was stirred at 0 °C for 15 min. After that, it was added to a mixture of the corresponding isocyanate S7 (1 equiv.) in anhydrous DCM (0.08 M) under a N atmosphere at 0 °C. The resulting mixture was stirred from 0 °C to room temperature for 30 min. The mixture was then poured into saturated NaHCO solution and extracted with DCM (×3). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give pure compound S8.

[0290] Synthesis of Example 346 [ka] Step 1: Synthesis of 346-2 To a mixture of 346-1 (150 mg, 0.688 mmol) and TEA (278 mg, 2.752 mmol) in anhydrous THF (10 mL) was added TMSNCO (119 mg, 1.032 mmol) dropwise under a N atmosphere at 0 °C, and the resulting mixture was stirred at room temperature for 16 h. The mixture was then poured into HO (50 mL) and washed twice with EtOAc (40 mL). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM:MeOH = 100:0 to 20:1) to give compound 346-2 (150 mg, 83.3% yield) as a white solid. LC / MS (ESI) m / z: 162 (M-100+H). + .

[0291] Step 2: Synthesis of 346-3 To a solution of 346-2 (180 mg, 0.575 mmol) in DCM (5 mL) was added TFA (1 mL). The resulting mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under reduced pressure to give crude compound 346-3 (158 mg, 99.0% yield) as a colorless oil, which was used in the next step without further purification. LC / MS (ESI) m / z: 162.2 (M+H) + .

[0292] Step 3: Synthesis of 346-4 To a solution of 346-3 (158 mg, 0.575 mmol) in anhydrous DCM (15 mL) was added 2,4-dimethoxybenzaldehyde (115 mg, 0.693 mmol) and acetic acid (69 mg, 1.155 mmol). The mixture was stirred at room temperature for 1 h, and then NaBH(OAc) (244 mg, 1.155 mmol) was added to the mixture, and the resulting mixture was stirred at 45 °C overnight. The mixture was then poured into 5% Na2CO3 solution (50 mL) and extracted with DCM (30 mL × 4). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH = 100:0 to 12:1) to give compound 346-4 (176 mg, 97.8% yield) as a colorless oil. LC / MS (ESI) m / z: 312 (M+H) + .

[0293] Step 4: Synthesis of 346-5 To a mixture of 346-4 (176 mg, 0.566 mmol) and acetic acid (348 mg, 5.788 mmol) in THF (5 mL) and EtOH (10 mL), (1-ethoxycyclopropoxy)trimethylsilane (111 mg, 0.637 mmol) was added, followed by NaBHCN (109 mg, 1.736 mmol). The resulting mixture was stirred at 80 °C for 16 h under a N atmosphere. The mixture was then poured into 5% NaCO solution (50 mL) and extracted with DCM (30 mL × 3). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH = 100:0 to 20:1) to give compound 346-5 (193 mg, 97.5% yield) as a colorless oil. LC / MS (ESI) m / z: 352 (M+H) + .

[0294] Step 5: Synthesis of 346-6 346-5 (193 mg, 0.55 mmol) was added to TFA (5 mL), and the resulting mixture was stirred at 80 °C for 16 h. The mixture was then concentrated to dryness under reduced pressure. The crude product was purified by flash column chromatography (eluted with DCM / MeOH = 100:0 to 15:1) to give compound 346-6 (78 mg, 70.9% yield) as a colorless oil. LC / MS (ESI) m / z: 202 (M+H) + .

[0295] Step 6: Synthesis of 346 To a solution of 346-6 (78 mg, 0.388 mmol) in anhydrous DCM (10 mL) was added DIEA (150 mg, 1.164 mmol), and the resulting mixture was stirred at 0 °C for 15 min. After that, a mixture of 2-fluoro-1-(isocyanatomethyl)-4-(trifluoromethoxy)benzene (92 mg, 0.388 mmol) in anhydrous DCM (5 mL) was added at 0 °C under a N atmosphere. The resulting mixture was stirred from 0 °C to room temperature for 30 min. The mixture was then poured into saturated NaHCO solution (50 mL) and extracted with DCM (30 mL × 3). The combined organic layer was dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give pure 346 (27 mg, 16.0% yield) as a white solid. LC / MS (ESI) m / z: 437.2 (M+H) + . 1 H NMR (400 MHz, MeOD-d4) δ 7.43 (t, J = 8.6 Hz, 1H), 7.09 (t, J = 7.8 Hz, 2H), 7.00 (t, J = 5.7 Hz, 1H), 4.59 - 4.37 (m, 3H), 4.30 (d, J = 12.6 Hz, 1H), 3.84 (d, J = 12.6 Hz, 1H), 3.69 (t, J = 11.9 Hz, 1H), 3.18 (t, J = 12.1 Hz, 1H), 2.72 - 2.63 (m, 1H), 2.59 - 2.53 (m, 1H), 2.37 - 2.24 (m, 2H), 1.01 - 0.92 (m, 2H), 0.83 - 0.73 (m, 2H).

[0296] General Procedure [ka] Step 1: Synthesis of T2 To a mixture of the corresponding amine T1 (1 equivalent) in a mixture of MeCN / DMF 5:1 (0.4 M), CDI (1 equivalent) was added at room temperature, and the resulting mixture was stirred for 2 h. The mixture was then concentrated under reduced pressure to give crude T2, which was used in the next step without further purification.

[0297] Step 2: Synthesis of T4 To a mixture of T2 (0.5 equiv.) and the appropriate T3 (1 equiv.) in MeCN (0.15 M) was added TEA (4 equiv.), and the resulting mixture was stirred at 50° C. for 16 h. The mixture was then concentrated to dryness, and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM:MeOH) to give T4.

[0298] Steps 3 to 7 are the same as steps 2 to 6 of the general procedure S'.

[0299] Synthesis of Example 347 [ka] Step 1: Synthesis of 347-1 To a mixture of N-methylamine hydrochloride (310 mg, 4.591 mmol) in a mixture of MeCN (10 mL) and DMF (2 mL) was added CDI (744 mg, 4.588 mmol) at room temperature, and the resulting mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under reduced pressure to give crude 347-1 (550 mg, 95.74% yield) as a pale yellow oil, which was used in the next step without further purification. LC / MS (ESI) m / z: 126 (M+H) + .

[0300] Step 2: Synthesis of 347-2 To a mixture of 347-1 (550 mg, 4.395 mmol) and tert-butyl ((3R,5S)-5-fluoropiperidin-3-yl)carbamate (480 mg, 2.199 mmol) in MeCN (15 mL) was added TEA (888 mg, 8.796 mmol), and the resulting mixture was stirred at 50 °C for 16 h. The mixture was then concentrated to dryness, and the residue was dissolved in EtOAc (40 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was then dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM:MeOH = 100:0 to 20:1) to give 347-2 (302 mg, 49.55% yield) as a colorless oil. LC / MS (ESI) m / z: 176 (M-100+H) + .

[0301] Steps 3 to 7 are the same as steps 2 to 6 of the general procedure S'.

[0302] Compound 347 was obtained as a white solid. LC / MS (ESI) m / z: 406.2 (M+H) + . 1 H NMR (400 MHz, MeOD) δ 7.43 (d, J = 9.6 Hz, 1H), 7.34 (d, J = 7.1 Hz, 1H), 7.04 (t, J = 5.9 Hz, 1H), 4.56 - 4.41 (m, 3H), 4.27 (d, J = 12.4 Hz, 1H), 3.81 (d, J = 12.6 Hz, 1H), 3.66 (t, J = 11.0 Hz, 1H), 3.17 - 3.08 (m, 1H), 2.69 (s, 3H), 2.68 - 2.62 (m, 1H), 2.57 (dt, J = 10.1, 3.4 Hz, 1H), 2.49 (s, 3H), 2.30 (m, 2H), 0.98 - 0.94 (m, 2H), 0.81 - 0.76 (m, 2H).

[0303] General Procedure U [ka] U1 was synthesized according to procedures P or Q by deprotecting intermediates P5 and Q3, respectively.

[0304] Step 1: Composite of U2 To a mixture of compound U1 (11.5 g, 39.59 mmol) and TEA (27.5 mL, 197.95 mmol) in anhydrous THF (200 mL) was added isocyanatotrimethylsilane (11.98 g, 83.15 mmol) dropwise at 0°C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 16 h. The mixture was then concentrated to dryness under reduced pressure. The residue was diluted with EtOAc (300 mL) and washed with water and brine. The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluting with DCM:MeOH = 100:0 to 95:5) to give compound U2 (10.0 g, 75.7% yield) as a pale yellow solid. LC / MS (ESI) m / z: 334 (M+H) + .

[0305] Step 2: Composite U3 Compound U2 (10 g, 29.99 mmol) was added portionwise to TFA (100 mL) under a N atmosphere at 0° C., and the resulting mixture was then stirred at 80° C. for 4 h. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure to give crude compound U3 (5.48 g, 99.8% yield) as a purple oil, which was used in the next step without further purification. LC / MS (ESI) m / z: 184 (M+H) + .

[0306] Step 3: Synthesis of U5 To a solution of U3 (1 equiv.) in anhydrous DCM (0.2 M) was added a mixture of the appropriate isocyanate U4 (1 equiv.) and TEA (10 equiv.) in DCM (0.15 M) dropwise at 0 °C under a N atmosphere. The resulting mixture was then stirred at room temperature for 1 h and concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc and washed with water and brine. The organic layer was then dried over anhydrous NaSO, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluting with DCM:MeOH) to give U5.

[0307] Synthesis of Example 352 [ka] To a solution of U3 (22.4 mg, 0.095 mmol) in anhydrous DCM (0.4 mL) was added a solution of 2-fluoro-1-(isocyanatomethyl)-4-(trifluoromethoxy)benzene (17.4 mg, 0.095 mmol) and TEA (0.13 mL, 0.95 mmol) in DCM (0.6 mL) dropwise under a N atmosphere at 0 °C. The resulting mixture was stirred at room temperature for 1 h and then concentrated to dryness under reduced pressure. The residue was dissolved in EtOAc (5 mL) and washed with water and brine. The organic layer was then dried over anhydrous NaSO, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 95:5) to give 352 (15 mg, 37.7% yield) as a white solid. LC / MS (ESI) m / z: 419.2 (M+H) + . 1H NMR (400 MHz, MeOD) δ 7.43 (t, J = 8.6 Hz, 1H), 7.09 (t, J = 7.9 Hz, 2H), 6.93 (d, J = 5.4 Hz, 1H), 4.43 (s, 2H), 4.02-3.86 (m, 2H), 3.69 - 3.59 (m, 1H), 3.18 (t, J = 12.0 Hz, 1H), 2.67 (t, J = 12.0 Hz, 1H), 2.59 - 2.49 (m, 1H), 2.18 - 2.05 (m, 1H), 1.94-1.84 (m, 1H), 1.80-1.71 (m, 1H), 1.59 - 1.45 (m, 1H), 1.00 - 0.89 (m, 2H), 0.82 - 0.71 (m, 2H).

[0308] General Procedure V: [ka] V3 was synthesized using procedures M or N.

[0309] Step 1: Synthesis of V2 To a solution of the appropriate V1 (1 eq.) in THF (0.3 M) was added TEA (3 eq.) and CDI (1.2 eq.), and the resulting mixture was stirred at room temperature for 12 h. The mixture was then concentrated to dryness under reduced pressure. The residue was dissolved in ethyl acetate and washed with water and brine. The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with PE: EtOAc) to give compound V2.

[0310] Step 2: Composite V4 Compound V2 (1.5 equivalents) was added to a mixture of compound V3 (1 equivalent) and TEA (3 equivalents) in DCM (0.05 M), and the resulting mixture was stirred at 50° C. for 16 hours. The mixture was then concentrated to dryness under reduced pressure, and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was then dried over anhydrous NaSO, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with PE:EtOAc) to give compound V4.

[0311] Synthesis of Example 348 [ka] Step 1: Synthesis of 348-1 To a solution of 2-((tert-butyldimethylsilyl)oxy)ethan-1-amine (50 mg, 0.29 mmol) in THF (1 mL) were added TEA (86.5 mg, 0.86 mmol) and CDI (55 mg, 0.34 mmol), and the resulting mixture was stirred at room temperature for 12 h. The mixture was then concentrated to dryness under reduced pressure, and the residue was dissolved in ethyl acetate (5 mL) and washed with water (10 mL) and brine (10 mL). The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with PE:EtOAc = 100:0 to 4:1) to give compound 348-1 (61 mg, 80.26% yield) as a white solid. LC / MS (ESI) m / z: 270 (M+H) + .

[0312] Step 2: Synthesis of 348-3 Compound 348-1 (16 mg, 0.06 mmol) was added to a mixture of compound 348-2 (15 mg, 0.04 mmol) and TEA (12 mg, 0.12 mmol) in DCM (0.7 mL), and the resulting mixture was stirred at 50 °C for 16 h. The mixture was then concentrated to dryness under reduced pressure, and the residue was dissolved in EtOAc (6 mL) and washed with water and brine. The organic layer was then dried over anhydrous NaSO, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluted with PE:EtOAc = 100:0 to 1:1) to give compound 348-3 (19.5 mg, 82.29% yield) as a yellow oil. LC / MS (ESI) m / z: 577 (M+H) + .

[0313] Step 6: Synthesis of 348 Compound 348-3 (19.5 mg, 0.034 mmol) was added dropwise to HCl / dioxane (0.5 mL, 4 M) at 0 °C, and the resulting mixture was stirred at room temperature for 1 h. The mixture was then concentrated under reduced pressure, and the residue was diluted with DCM / MeOH (15:1, 1 mL). Saturated NaHCO3 solution was slowly added to bring the pH to 8-9, and the organic layer was then separated, washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by column chromatography on silica gel (eluted with DCM:MeOH = 100:0 to 20:1) to give 348 (15 mg, 96.2% yield) as a yellow oil. LC / MS (ESI) m / z: 463.3 (M+H) + . 11H NMR (400 MHz, MeOD) δ 7.43 (t, J = 8.1 Hz, 1H), 7.08 (d, J = 9.3 Hz, 2H), 4.51 - 4.34 (m, 2H), 3.94 (dd, J = 30.8, 9.7 Hz, 2H), 3.62 (d, J = 25.8 Hz, 3H), 3.28 (s, 2H), 3.14 (s, 1H), 2.70 - 2.66 (m, 1H), 2.55 - 2.51 (m, 1H), 2.11 (d, J = 11.1 Hz, 1H), 1.94 - 1.90 (m, 1H), 1.77 - 1.73 (m, 1H), 1.53 (s, 1H), 0.97 - 0.93 (m, 2H), 0.76 (brs, 2H).

[0314] Examples 344 to 370

Table 20

[0315] Examples 371 to 377 The compounds listed in the table below were prepared by applying the experimental general procedures G, R', and S described above. [Table 21] TIFF0007797723000237.tif223167 TIFF0007797723000238.tif72169

[0316] Examples 378-400 listed in the table below were prepared by applying the experimental general procedures W, X, or Y listed after the table. [Table 22] TIFF0007797723000240.tif241170 TIFF0007797723000241.tif236170 TIFF0007797723000242.tif226170 TIFF0007797723000243.tif237170 TIFF0007797723000244.tif250163 TIFF0007797723000245.tif249166 TIFF0007797723000246.tif237170 TIFF0007797723000247.tif231170 TIFF0007797723000248.tif232170 TIFF0007797723000249.tif211170

[0317] General steps: [ka] Step 1: Synthesis of W2 To a mixture of the appropriate aldehyde W1 (1 equivalent) and hydroxylamine hydrochloride (1.1 equivalents) in DCM was added TEA (1.1 equivalents) dropwise at 0°C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 45 minutes. The mixture was then diluted with water and extracted twice with DCM. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to give crude compound W2, which was used in the next step without further purification.

[0318] Step 2: Synthesis of W4 To a mixture of W2 (1 equivalent) and W3 (1 equivalent) in MeOH / HO (4:1, V / V) was added [bis(trifluoroacetoxy)iodo]benzene (1.3 equivalents) under a N2 atmosphere at 0°C. The resulting mixture was stirred at room temperature for 1 hour. The mixture was then diluted with water and extracted with EtOAc. The combined organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with PE / EtOAc) to give compound W4.

[0319] Step 3: Synthesis of W5 W4 (1 equivalent) was added dropwise to a mixture of TFA / DCM (1:4, V / V) at 0° C., and the resulting mixture was stirred at room temperature for 1 hour. The mixture was then concentrated to dryness under reduced pressure to give crude compound W5, which was used in the next step without further purification.

[0320] Step 4: Synthesis of W To a mixture of W5 (1 eq.) and NaHCO3 (3 eq.) in DCM was added a solution of triphosgene (0.5 eq.) in DCM at -30°C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 30 minutes, and then a mixture of W6 (1 eq.) and TEA (3 eq.) in anhydrous DCM was added dropwise at 0°C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 1 hour, and then the mixture was diluted with water and extracted twice with DCM. The combined organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give pure compound W.

[0321] Synthesis of Example 400: [ka] Step 1: Synthesis of W2 To a solution of W1 (1.0 g, 5.26 mmol) and hydroxylamine hydrochloride (402 mg, 5.79 mmol) in DCM (25 mL) was added TEA (586 mg, 5.79 mmol) dropwise under a N atmosphere at 0 °C, and the resulting mixture was stirred at room temperature for 45 min. The mixture was then diluted with water (25 mL) and extracted twice with DCM (25 mL). The combined organic layer was separated, washed with brine (30 mL), dried over anhydrous NaSO, filtered, and concentrated to dryness under reduced pressure to give crude compound W2 (950 mg, 88.05% yield), which was used in the next step without further purification. LC / MS (ESI) m / z: 206 (M+H) + .

[0322] Step 2: Synthesis of W4 To a mixture of W2 (950 mg, 4.63 mmol) and W3 (719 mg, 4.63 mmol) in MeOH (40 mL) and HO (10 mL) was added [bis(trifluoroacetoxy)iodo]benzene (2.59 g, 6.02 mmol) under a N2 atmosphere at 0 °C, and the resulting mixture was stirred at room temperature for 1 h. The mixture was then diluted with water (60 mL) and extracted twice with EtOAc (50 mL). The combined organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with 10–50% EtOAc / PE) to give pure compound W4 (390 mg, 23.50% yield) as a white solid. LC / MS (ESI) m / z: 359 (M+H) + .

[0323] Step 3: Synthesis of W5 To a mixture of W4 (390 mg, 1.09 mmol) in DCM (4 mL) was added TFA (1 mL) under N2 atmosphere at 0 °C, and the resulting mixture was stirred at room temperature for 1 h. The mixture was then concentrated to dryness under reduced pressure to give crude product W5 (550 mg), which was used in the next step without further purification. LC / MS (ESI) m / z: 259 (M+H) + .

[0324] Step 4: Synthesis of 400 A solution of triphosgene (35 mg, 0.54 mmol) in DCM (2 mL) was added dropwise to a solution of crude W5 (60 mg, 0.232 mmol) and NaHCO (59 mg, 0.697 mmol) in DCM (5 mL) at −30° C. under a N atmosphere, and the resulting mixture was stirred at room temperature for 30 min. Then, a mixture of W6 (43 mg, 0.232 mmol) and TEA (71 mg, 0.697 mmol) in anhydrous DCM (5 mL) was added dropwise at 0° C. under a N atmosphere, and the resulting mixture was stirred at room temperature for 1 h. The mixture was then diluted with water (10 mL) and extracted twice with DCM (10 mL). The combined organic layer was separated, washed with brine (12 mL), dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give 400 (22 mg, 20.25% yield) as a white solid. LC / MS: m / z 468 (M+H) + . 1H NMR (400 MHz, MeOD) δ 7.86 - 7.77 (m, 1H), 7.74 (s, 1H), 7.59 (t, J = 8.0 Hz, 1H), 7.43 - 7.31 (m, 1H), 6.71 (s, 1H), 4.59 - 4.44 (m, 2H), 4.05 - 3.84 (m, 2H), 3.72 - 3.56 (m, 1H), 3.24 - 3.14 (m, 1H), 2.75 - 2.61 (m, 1H), 2.58 - 2.48 (m, 1H), 2.22 - 2.05 (m, 1H), 1.97 - 1.84 (m, 1H), 1.81 - 1.68 (m, 1H), 1.61 - 1.45 (m, 1H), 1.03 - 0.90 (m, 2H), 0.88 - 0.75 (m, 2H).

[0325] General Procedure X: [ka] Step 1: Merge X2 To a solution of X1 (1 equiv.) and diethyl oxalate (1.1 equiv.) in anhydrous THF was added NaH (1.1 equiv.) under a N2 atmosphere at 0 °C, and the resulting mixture was stirred at room temperature for 3 h. The mixture was then quenched with water and extracted twice with DCM. The combined organic layer was separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to give crude compound X2, which was used in the next step without further purification.

[0326] Step 2: Merge X3 Hydroxylamine hydrochloride (2.5 equiv.) was added to a solution of X2 (1 equiv.) in EtOH, and the resulting mixture was stirred at 80° C. under N2 atmosphere for 2 h. After cooling, the mixture was concentrated to dryness under reduced pressure, and the residue was purified by flash column chromatography (eluted with PE / EtOAc) to give compound X3.

[0327] Step 3: Merge X4 To a solution of compound X3 (1 equivalent) in EtOH, NaBH4 (2.5 equivalents) was added portionwise at 0 °C, and the resulting mixture was stirred at room temperature for 2 hours. The mixture was then quenched with water and extracted twice with EtOAc. The combined organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluted with PE / EtOAc) to give compound X4.

[0328] Step 4: Merge X5 TEA (2 equivalents) and DPPA (1.2 equivalents) were added to a solution of compound X4 (1 equivalent) in toluene, and the resulting mixture was stirred at room temperature under a N atmosphere for 20 hours. The mixture was then diluted with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated to dryness under reduced pressure to give crude product X5, which was used in the next step without further purification.

[0329] Step 5: Merge X6 PPh3 (2 equiv.) was added to a mixture of X5 (1 equiv.) in THF / HO (4:1, V / V), and the resulting mixture was stirred at room temperature under a N2 atmosphere for 17 hours. The mixture was then diluted with 2N HCl (aq.) and washed with MTBE. The aqueous layer was separated, and the pH was adjusted to 8 with saturated aqueous NaHCO3, followed by extraction twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to give crude product X6, which was used in the next step without further purification.

[0330] Step 6: Composition of X To a mixture of X6 (1 equivalent) and NaHCO3 (3 equivalents) in anhydrous DCM, a solution of triphosgene (0.5 equivalents) in DCM was added dropwise at -30°C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 30 minutes. Then, a mixture of X7 (1 equivalent) and TEA (3 equivalents) in anhydrous DCM was added dropwise at 0°C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 1 hour. The mixture was then diluted with water and extracted twice with DCM. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give compound X.

[0331] General Procedure: [ka] Step 1: Synthesis of Y2 To a solution of Y1 (1 equiv.) in MeCN was added t-BuONO (1.2 equiv.) and TMSN3 (1.2 equiv.) under a N2 atmosphere at 0 °C. The resulting mixture was stirred at room temperature for 1 h. When TLC showed complete consumption of the starting material, the solution of Y2 in MeCN was used directly in the next step without further purification.

[0332] Step 2: Synthesis of Y4 To a mixture of Y3 (1.2 equiv.), sodium ascorbate (0.2 equiv.), and CuSO4 (0.2 equiv.) in MeCN was added a solution of Y2 (1 equiv.) in MeCN dropwise at 0 °C under a N2 atmosphere. The resulting mixture was stirred overnight at room temperature under a N2 atmosphere. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with PE / EtOAc) to give Y4.

[0333] Step 3: Synthesis of Y5 Y4 (1 equivalent) was added dropwise to a mixture of TFA in DCM (1:3, V / V) at 0° C. The resulting mixture was stirred at room temperature for 30 minutes. The mixture was then concentrated to dryness. The residue was dissolved in EtOAc, neutralized with aqueous NaHCO3, and extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to give crude 5, which was used directly in the next step without further purification.

[0334] Step 4: Synthesis of Y7 To a mixture of Y5 (1.0 equiv.) and Y6 (1.0 equiv.) in DCM was added dropwise a solution of TEA (3.0 equiv.) and BTC (0.5 equiv.) in DCM at −78° C. under a N atmosphere. The resulting mixture was stirred at room temperature for 1 hour. The mixture was then diluted with water and extracted twice with DCM. The combined organic layer was washed with brine, dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give Y7.

[0335] Synthesis of Example 386: [ka] Step 1: Synthesis of Y2 To a solution of Y1 (3.0 g, 16.95 mmol) in MeCN (30 mL) was added t-BuONO (2.10 g, 20.34 mmol) and TMSN3 (2.34 g, 20.34 mmol) dropwise under a N2 atmosphere at 0 °C. The resulting mixture was stirred at room temperature for 1 h. When TLC showed complete consumption of the starting material, the solution of Y2 in MeCN was used directly in the next step without further purification.

[0336] Step 2: Synthesis of Y4 To a mixture of Y3 (3.15 g, 20.34 mmol), sodium ascorbate (810 mg, 4.07 mmol), and CuSO4 (650 mg, 4.07 mmol) in MeCN (20 mL) was added a mixture of Y2 in MeCN (30 mL) dropwise at 0 °C. The resulting mixture was stirred overnight at room temperature under a N2 atmosphere. The mixture was then diluted with water and extracted twice with EtOAc (40 mL). The combined organic layers were washed with brine (40 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with PE / EtOAc = 100:0 to 3:2) to give pure Y4 (2.3 g, 37.80% yield). LC / MS (ESI) m / z: 359 (M+H) + .

[0337] Step 3: Synthesis of Y5 To a solution of Y4 (80 mg, 0.223 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise at 0° C. The resulting mixture was stirred at room temperature for 30 minutes. Then, the mixture was concentrated to dryness. The residue was dissolved in EtOAc (4 mL) and neutralized with aqueous NaHCO to pH=8. The mixture was extracted twice with EtOAc (10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous NaSO, filtered, and concentrated to give crude Y5 (55 mg, 96.49% yield), which was used directly in the next step without further purification. LC / MS (ESI) m / z: 259 (M+H) + .

[0338] Step 4: Synthesis of Example 386 To a mixture of Y5 (55 mg, 0.212 mmol) and Y6 (46 mg, 0.212 mmol) in DCM (5 mL) was added dropwise a solution of TEA (64 mg, 0.636 mmol) and BTC (31 mg, 0.106 mmol) in DCM (1 mL) at −78° C. under a N atmosphere. The resulting mixture was stirred at room temperature for 1 h. The mixture was then diluted with water (20 mL) and extracted twice with DCM (20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous NaSO, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give 386 (15 mg, 9.74% yield) as a white solid. LC / MS (ESI) m / z: 500 (M+H) + . 1 HNMR (400 MHz, MeOD) δ 8.43 (s, 1H), 7.88 (dd, J = 12.0, 3.9 Hz, 2H), 7.69 (t, J = 8.2 Hz, 1H), 7.42 (d, J = 8.3 Hz, 1H), 4.52 (s, 2H), 4.52 - 4.47 (m, 1H), 4.44 - 4.36 (m, 1H), 4.29 (d, J = 12.5 Hz, 1H), 3.82 (d, J = 12.8 Hz, 1H), 3.71 (t, J = 11.1 Hz, 1H), 3.18 - 3.08 (m, 1H), 2.69 (s, 3H), 2.68 - 2.62 (m, 1H), 2.57 - 2.51 (m, 1H), 2.38 - 2.31 (m, 1H), 2.30 - 2.19 (m, 1H), 0.99 - 0.91 (m, 2H), 0.83 - 0.76 (m, 2H).

[0339] Representative compounds selected from above were tested in the isoleucine transport assay described in Example 1. The results are summarized in the tables of FIGS.

[0340] Incorporation by Reference All US patents and US patent application publications cited herein are hereby incorporated by reference.

[0341] equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims. [Section 1] Compounds of formula (I): [ka] (In the formula, n is 0, 1, or 2; L1 is absent or selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH2-; L2 is absent or is -CH2-; L3 is absent or is -C(O)-; X1 and X2 are independently selected from -H, alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl, with the proviso that X1 and X2 are not both -H; Y1 is selected from aryl and heteroaryl; Y2 is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NH(Y2'), and -N(Y2'')2; Y2' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, and cycloalkyl; each Y2" is alkyl, or both occurrences together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocyclyl; Y3, Y4, Y5, and Y6 are independently selected from -H, -OH, halide, alkyl, haloalkyl, and alkoxy, with the proviso that Y3 and Y4 or Y5 and Y6 are not both -OH; provided that when L3 is -C(O)-, Y2 is not aryl and the compound is [ka] (not selected from or a pharmaceutically acceptable salt thereof. [Section 2] Item 1. The compound according to item 1, wherein Y2' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, and cycloalkyl. [Section 3] The compound according to item 1 or 2, wherein one of X1 and X2 is -H, and the other of X1 and X2 is selected from C1-C4 alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl. [Section 4] One of X1 and X2 is -H, and the other of X1 and X2 is -CH3, -CH2CH3, -CH2CF3, -CH2CH2CH3, [ka] Item 4. The compound according to item 3, selected from: [Section 5] Item 5. The compound according to item 4, wherein X1 is -H and X2 is -CH3. [Section 6] Item 5. The compound according to item 4, wherein X2 is -H and X1 is -CH3. [Section 7] X1 is -H and X2 is [ka] Item 5. The compound according to item 4, wherein [Section 8] X2 is -H and X1 is [ka] Item 5. The compound according to item 4, wherein [Section 9] Item 9. The compound according to any one of items 1 to 8, wherein L1 is absent. [Section 10] Item 9. The compound according to any one of items 1 to 8, wherein L1 is selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH2-. [Section 11] Item 11. The compound according to item 10, wherein L1 is selected from -CH2-, -C(H)(CH3)-, -CH2CH2-, and -C(H)(OH)CH2-. [Section 12] L1 is, [ka] Item 11. The compound according to item 10, wherein [Section 13] L1 is, [ka] Item 13. The compound according to item 12, selected from: [Section 14] L1 is, [ka] Item 11. The compound according to item 10, selected from: [Section 15] The compound according to any one of items 1, 2 and 9-14, having a structure selected from the following: [ka] [Section 16] Item 16. The compound according to any one of items 1 to 15, wherein Y1 is unsubstituted aryl. [Section 17] Item 17. The compound according to item 16, wherein Y1 is selected from unsubstituted phenyl and unsubstituted naphthyl. [Section 18] Item 16. The compound according to any one of items 1 to 15, wherein Y1 is substituted aryl. [Section 19] Y1, [ka] and R1, R2, R3, R4, and R5 are each independently -H, halogen, -CN, 、 The compound according to item 18, wherein R1, R2, R3, R4, and R5 are independently selected from -CF3, -CHF2, -CF2CH3, -OCF3, -OCHF2, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl, with the proviso that one of R1, R2, R3, R4, and R5 is not -H. [Section 20] R1, R2, R3, R4, and R5 are -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -OCH3, -OCF3, -OCHF2, [ka] 20. The compound according to item 19, wherein the compound is independently selected from: [Section 21] R1, R2, R3, R4, and R5 are -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH 3、 -OCF3, and [ka] 20. The compound according to item 19, wherein the compound is independently selected from: [Section 22] 22. The compound according to any one of items 19 to 21, wherein two of R1, R2, R3, R4, and R5 are not —H. [Section 23] 22. The compound according to any one of items 19 to 21, wherein three of R1, R2, R3, R4, and R5 are not —H. [Section 24] Y1, [ka] Item 20. The compound according to item 19, selected from: [Section 25] Y1, [ka] Item 20. The compound according to item 19, selected from: [Section 26] Item 16. The compound according to any one of items 1 to 15, wherein Y1 is unsubstituted heteroaryl. [Section 27] Y1, [ka] Item 24. The compound according to item 23, selected from: [Section 28] Item 16. The compound according to any one of items 1 to 15, wherein Y1 is substituted heteroaryl. [Section 29] Y1, [ka] is selected from 29. The compound of claim 28, wherein each occurrence of R6, R7, R8, and R9 is independently selected from -H, halogen, -CN, -OCF3, -OCHF2, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, aryl, and heteroaryl, provided that at least one of R6, R7, R8, and R9 is not -H. [Section 30] Item 30. The compound according to any one of items 1 to 29, wherein L2 is absent. [Section 31] Item 30. The compound according to any one of items 1 to 29, wherein L2 is -CH2-. [Section 32] Item 32. The compound according to any one of items 1 to 31, wherein L3 is absent. [Section 33] Item 32. The compound according to any one of items 1 to 31, wherein L3 is -C(O)-. [Section 34] Item 34. The compound according to any one of items 1 to 33, wherein n is 0. [Section 35] Item 34. The compound according to any one of items 1 to 33, wherein n is 1. [Section 36] Item 34. The compound according to any one of items 1 to 33, wherein n is 2. [Section 37] The compound of any one of paragraphs 1, 32, and 34, having a structure selected from: [ka] [Section 38] The compound of any one of paragraphs 1, 32, and 35, having a structure selected from the following: [ka] [Section 39] The compound of any one of paragraphs 1, 32, and 36, having a structure selected from the following: [ka] [Section 40] Item 40. The compound according to any one of items 37 to 39, wherein Y2 is unsubstituted heteroaryl. [Section 41] Y2 is, [ka] 41. The compound according to item 40, selected from: [Section 42] Y2 is, [ka] Item 42. The compound according to item 41, wherein [Section 43] Item 40. The compound according to any one of items 37 to 39, wherein Y2 is substituted heteroaryl. [Section 44] Y2 is, [ka] and R 10 , R 11 , and R 12 -H, halogen, -CN, -OH 、-NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 -CO2R 15 , and -C(O)NHSO2R 15 are independently selected from, where R 10 , R 11 , and R 12 At least one of the is not -H, R 13 , R 14 , and R 15 44. The compound of clause 43, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl. [Section 45] R 10 , R 11 , and R 12 -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH 3、 -CF3, -CHF2, -CF2CH 3、 45. The compound according to claim 44, independently selected from -OCH3, -OCF3, -OCHF2, -OAc, -NH2, -NHCH3, -NHAc, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, -C(O)NHSO2CH3, -C(O)NHSO2CH2CH3, -CO2H, phenyl, cyclopropyl, cyclobutyl, imidazolyl, and tetrazolyl. [Section 46] R 10 and R 12 are -H and R, respectively. 11 is -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, [ka] 46. ​​The compound according to claim 45, selected from: [Section 47] R 11 and R 12are -H and R, respectively. 10 is -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, [ka] 46. ​​The compound according to claim 45, selected from: [Section 48] R 10 and R 11 are -H and R, respectively. 12 is -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3, [ka] 46. ​​The compound according to claim 45, selected from: [Section 49] Y2 is, [ka] is selected from R 16 is, for each occurrence, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14 , -CO2R 15 are independently selected from R 13 , R 14 , and R 15 44. The compound of clause 43, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl. [Section 50] R 16 -CN, -CH3, -CF3, -C(O)NH2, -CO2CH2CH3, and [ka] Item 50. The compound according to item 49, selected from: [Section 51] Y2 is, [ka] is selected from R 17 , R 18 , R 19 , R 20 , and R 21 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from, where R 17 , R 18 , R 19 , R 20 , and R 21 At least one of the is not -H, R 13 , R 14 , and R 15 44. The compound of clause 43, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl. [Section 52] R 17 , R 18 , R 19 , R 20 , and R 21 52. The compound according to clause 51, wherein is independently selected from -H, -CN, -CH3, and -OCH3. [Section 53] Y2 is, [ka] 44. The compound according to claim 43, selected from: [Section 54] The compound of any one of paragraphs 1, 33, and 34, having a structure selected from the following: [ka] [Section 55] 36. The compound of any one of paragraphs 1, 33, and 35, having a structure selected from: [ka] [Section 56] 37. The compound of any one of paragraphs 1, 33, and 36, having a structure selected from: [ka] [Section 57] 57. The compound according to any one of items 54 to 56, wherein Y2 is unsubstituted cycloalkyl or heterocyclyl. [Section 58] Y2 is, [ka] Item 58. The compound according to item 57, selected from: [Section 59] Y2 is, [ka] Item 58. The compound according to item 57, selected from: [Section 60] 57. The compound according to any one of items 54 to 56, wherein Y2 is substituted cycloalkyl or heterocyclyl. [Section 61] Y2 is, [ka] 61. The compound according to claim 60, selected from: [Section 62] 57. The compound according to any one of items 54 to 56, wherein Y2 is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, and hydroxyalkyl. [Section 63] 63. The compound according to clause 62, wherein Y2 is selected from -CH3, -CH2CH(CH3)2, -CH2CH2C≡CH, -CH2CH2OCH3, -C(H)(CH3)CH2OCH3, -OCH3, -CH2OH, -CH2CH2OH, -C(CH3)2OH, and -CH2OCH3. [Section 64] 64. The compound according to clause 63, wherein Y2 is selected from -CH2OH and -CH2CH2OH. [Section 65] 57. The compound according to any one of items 54 to 56, wherein Y2 is heteroaryl. [Section 66] Y2 is, [ka] 66. The compound according to claim 65, selected from: [Section 67] Y2 is, [ka] and R 10 , R 11 , and R 12 -H, halogen, -CN, -OH 、 -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 66. The compound of clause 65, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl. [Section 68] R 10 , R 11 , and R 12 Item 68. The compound according to item 67, wherein at least one of: is not —H. [Section 69] Y2 is, [ka] is selected from R 17 , R 18 , R 19 , R 20 , and R 21 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 15 66. The compound of clause 65, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl. [Section 70] R 17 , R 18 , R 19 , R 20 , and R 21 Item 70. The compound according to item 69, wherein at least one of: is not —H. [Section 71] Y2 is, [ka] is selected from R 22 , R 23 , R 24 , and R 25 Each occurrence of is -H, halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14、 and -CO2R 15 are independently selected from R 13 , R 14 , and R 1566. The compound of clause 65, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl. [Section 72] R 22 , R 23 , R 24 , and R 25 72. The compound of clause 71, wherein each occurrence of is independently selected from -H and -CH3. [Section 73] 57. The compound according to any one of items 54 to 56, wherein Y2 is -NH(Y2') or Y2 is -N(Y2'')2. [Section 74] Y2' is -H, -OH, -OCH3, -CH3, -CH2CH2OCH3, and [ka] 74. The compound according to claim 73, selected from: [Section 75] 74. The compound according to paragraph 73, wherein each Y2'' is -CH3. [Section 76] 74. The compound according to paragraph 73, wherein both Y2'' together with the nitrogen atom to which they are attached form morpholinyl. [Section 77] 57. The compound according to any one of items 54 to 56, wherein Y2 is -NH(Y2'). [Section 78] 78. The compound according to clause 77, wherein Y2' is selected from -H, alkyl, alkoxy, and hydroxyalkyl. [Section 79] 79. The compound according to clause 78, wherein Y2' is selected from -H, -OCH3, -CH3, and -CH2CH2OH. [Section 80] Item 80. The compound according to any one of items 1 to 79, wherein Y3 and Y4 are both -H or both -F. [Section 81] Item 80. The compound according to any one of items 1 to 79, wherein Y3 is selected from -F, -CF3, -OH and -OCH3, and Y4 is -H. [Section 82] Item 80. The compound according to any one of items 1 to 79, wherein Y4 is selected from -F, -CF3, -OH and -OCH3, and Y3 is -H. [Section 83] Item 80. The compound according to any one of items 1 to 79, wherein Y5 and Y6 are both -H or both -F. [Section 84] Item 80. The compound according to any one of items 1 to 79, wherein Y5 is selected from -F, -CF3, -OH and -OCH3, and Y6 is -H. [Section 85] Item 80. The compound according to any one of items 1 to 79, wherein Y6 is selected from -F, -CF3, -OH and -OCH3, and Y5 is -H. [Section 86] A compound having the structure of any one of the following compounds or a pharmaceutically acceptable salt thereof: [Table 1] TIFF0007797723000296.tif251158 TIFF0007797723000297.tif241167 TIFF0007797723000298.tif244167 TIFF0007797723000299.tif251160 TIFF0007797723000300.tif251163 TIFF0007797723000301.tif251158 TIFF0007797723000302.tif251162 TIFF0007797723000303.tif250167 TIFF0007797723000304.tif240167 TIFF0007797723000305.tif251159 TIFF0007797723000306.tif251167 TIFF0007797723000307.tif251158 TIFF0007797723000308.tif212167 TIFF0007797723000309.tif238167 TIFF0007797723000310.tif248167 TIFF0007797723000311.tif251167 TIFF0007797723000312.tif246167 TIFF0007797723000313.tif240167 TIFF0007797723000314.tif250167 TIFF0007797723000315.tif250167 TIFF0007797723000316.tif228167 TIFF0007797723000317.tif251160 TIFF0007797723000318.tif241167 TIFF0007797723000319.tif227167 TIFF0007797723000320.tif252167 TIFF0007797723000321.tif245167 TIFF0007797723000322.tif251164 TIFF0007797723000323.tif251164 TIFF0007797723000324.tif235167[Section 87] A compound having the structure of any one of the following compounds or a pharmaceutically acceptable salt thereof: [Table 2] TIFF0007797723000326.tif251162 TIFF0007797723000327.tif208167[Section 88] A compound having the structure of any one of the following compounds or a pharmaceutically acceptable salt thereof: [Table 3] [Section 89] A compound having the structure of any one of the following compounds or a pharmaceutically acceptable salt thereof: [Table 4] TIFF0007797723000330.tif235167 TIFF0007797723000331.tif237167[Section 90] Item 89. A pharmaceutical composition comprising the compound according to any one of items 1 to 89 and a pharmaceutically acceptable excipient. [Section 91] A method for treating or preventing a disease or disorder associated with a genetic deficiency of phenylalanine hydroxylase, comprising administering to a subject in need thereof an effective amount of a compound according to any one of items 1 to 89. [Section 92] A method for treating or preventing phenylketonuria, comprising administering to a subject in need thereof an effective amount of the compound according to any one of items 1 to 89. [Section 93] A method for treating or preventing hyperphenylalaninemia, comprising administering to a subject in need thereof an effective amount of a compound according to any one of items 1 to 89. [Section 94] Item 94. The method of any one of Items 91 to 93, wherein the compound reduces phenylalanine levels in the subject's body. [Section 95] 90. A method for treating or preventing tyrosinemia (type I, II, or III), comprising administering to a subject in need thereof an effective amount of a compound according to any one of paragraphs 1 to 89. [Section 96] 96. The method of paragraph 95, wherein the compound reduces tyrosine levels in the subject's body. [Section 97] 89. A method for treating or preventing nonketotic hyperglycinemia, comprising administering to a subject in need thereof an effective amount of the compound according to any one of items 1 to 89. [Section 98] 98. The method of paragraph 97, wherein the compound reduces glycine levels in the subject's body. [Section 99] Item 89. A method for treating or preventing isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorders, or hyperammonemia, comprising administering to a subject in need thereof an effective amount of the compound according to any one of items 1 to 89. [Section 100] 89. 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, comprising administering to a subject in need thereof an effective amount of a compound according to any one of items 1 to 89. [Section 101] Item 101. The method of any one of Items 91 to 100, wherein the compound inhibits SLC6A19 in the subject.

Claims

1. Compounds of formula (I): 【Chemistry 1】 (In the formula, n is 1, L １ is —CH 2 —; L ２ is absent, L ３ is —C(O)—, X １ and X ２ is independently selected from —H and cyclopropyl, with the proviso that X １ and X ２ are not both -H, Y １ teeth, 【Chemistry 2】 and R 1 , R 2 , R 3 , R 4 , and R 5 are —H, —F, —Cl, —Br, —CN, —CH 3 , —CH 2 CH 3 , —OCF 3 , and 【Transformation 3】 are independently selected from Y ２ represents hydroxyalkyl having 1 to 3 carbon atoms and —NH(Y ２ ') are selected from Y ２ ' is alkyl having 1 to 3 carbon atoms, Y ３ , Y ４ , Y ５ , and Y ６ are independently selected from —H and halide. or a pharmaceutically acceptable salt thereof.

2. (A)Y １ が、 【Chemistry 4】 or (B) Y １ but, 【Transformation 5】 2. The compound of claim 1, selected from:

3. Y １ but, 【Transformation 6】 3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, selected from:

4. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein Y 2 is hydroxyalkyl having 1 to 3 carbon atoms.

5. The compound according to claim 3, or a pharmaceutically acceptable salt thereof, wherein Y 2 is hydroxyalkyl having 1 to 3 carbon atoms.

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