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 of U.S. Provisional Patent Application No. 63 / 308,790, filed on February 10, 2022; No. 63 / 292,815, filed on December 22, 2021; No. 63 / 234,487, filed on August 18, 2021; No. 63 / 226,551, filed on July 28, 2021; and No. 63 / 159,271, filed on March 10, 2021.

Background Art

[0002] Phenylketonuria (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, and the plasma concentration of phenylalanine becomes abnormally high. People with PKU have an abnormally high blood concentration of phenylalanine, and if untreated, it can cause irreversible nerve damage and various complications such as intellectual disability, seizures, neurodevelopmental disorders, and behavioral disorders. PKU is difficult to treat because the blood concentration of phenylalanine is directly related to diet. Patients must adhere to a strict diet throughout their lives, which affects all aspects of their lives. Current standard treatments are enzyme cofactor therapy and enzyme replacement therapy, but these therapies are not effective for all patients and are associated with a potential risk of adverse events.

[0003] The enzyme that metabolizes phenylalanine and thus plays a role in 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 types of PKU. These LOF mutations that cause PKU can be diagnosed as classical PKU (the most severe form) and less severe forms such as "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 necessary for PAH activity, can also increase phenylalanine concentrations. In addition to diet, blood amino acid concentrations, including phenylalanine concentration, are regulated by SLC6A19. SCL6A19 is located in the proximal renal tubules of the kidney and plays a role in reabsorbing amino acids and returning them to the bloodstream.

Summary of the Invention

Means for Solving the Problems

[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 by modulating SLC6A19 transport.

[0005] Accordingly, provided herein are compounds having the structure of formula (I):

Chemical

Chemical formula

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

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

[0009] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred 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. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0010] Other features, objects, and advantages of the present invention will become apparent from the description of the invention and the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0011]

Figure 1-1

Figure 1-2

Figure 1-3

Figure 2

Figure 3

Mode for Carrying Out the Invention

[0012] Definitions For reference, before further describing the present invention, specific terms employed in this specification, examples, and the appended claims are gathered herein. These definitions are to be construed in light of the remainder of the disclosure and understood by those skilled in the art. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art.

[0013] To more readily understand the present invention, specific terms and phrases are defined below and throughout the specification.

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

[0015] As used in this specification and the claims, the phrase "and / or" is to be understood to mean "either or both" of the recited elements, i.e., elements that may be conjunctively present in some cases and disjunctively present in other cases. The multiple elements recited using "and / or" are to be construed in the same fashion, i.e., as "one or more" of the recited elements so combined. Optionally, other elements may be present, whether or not related to the elements specifically recited by the "and / or" clause, depending on whether or not they are relevant to those specifically recited elements. Thus, by way of non-limiting example, reference to "A and / or B", when used in conjunction with open-ended language such as "comprising", may refer in one embodiment to only A (optionally including elements other than B), in another embodiment to only B (optionally including elements other than A), and in yet another embodiment to both A and B (optionally including other elements).

[0016] As used in this specification and the claims, "or" is to 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" is to be construed as inclusive, i.e., including not only one or at least one of the number of elements or items in the list, but also two or more, and optionally including additional items not in the list. Only terms that clearly indicate the contrary, such as "only one of" or "exactly one of", or "consisting of" when used in the claims, refer to exactly one element of the number of elements or items in the list. In general, the term "or" as used in this specification is to be construed as referring to exclusive alternatives (i.e., "either one or the other, but not both") only when preceded by terms indicating exclusivity such as "any one of", "one of", "only one of", or "exactly one of". "Consisting essentially of" is to have the ordinary meaning as used in the field of patent law when used in the claims.

[0017] As used in this specification and the claims, the phrase "at least one" shall be understood to mean at least one element selected from any one or more of the elements in a list of one or more elements, but it is not necessary to necessarily include at least one of each and every element specifically listed in the list of elements, and it does not exclude any combination of the elements in the list of elements. This definition also acknowledges that, regardless of whether related to specifically identified elements or not, elements other than those specifically identified in the list of elements referred to by the phrase "at least one" may optionally exist. Thus, by way of 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") may, in one embodiment, refer to including at least one, and optionally a plurality, of A and no presence of B (and optionally including elements other than B), in another embodiment, refer to including at least one, and optionally a plurality, of B and no presence of A (and optionally including elements other than A), and in yet another embodiment, may refer to including at least one, and optionally a plurality, of A and including at least one, and optionally a plurality, of B (and optionally including other elements), etc.

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

[0019] In the claims and the above specification, transitional phrases such as "comprising", "including", "carrying", "having", "containing", "involving", "holding", "composed of", etc. are all non-limiting, that is, it should be understood to mean including, but not limited to. As stipulated in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03, only the transitional phrases "consisting of" and "consisting essentially of" are restrictive or semi-restrictive transitional phrases, respectively.

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

[0021] "Geometric isomers" means isomers in which the orientation of substituent atoms is different with respect to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. The atoms (other than H) on both sides of a carbon-carbon double bond can be in the E configuration (substituents are on the opposite side of the carbon-carbon double bond) or the Z configuration (substituents are facing the same side). "R", "S", "S * ", "R *The terms ", "E", "Z", "cis", and "trans" indicate the configuration with respect to the core molecule. Some of the disclosed compounds may exist in "atropisomeric" forms or as "atropisomers". Atropisomers are stereoisomers that result from restricted rotation about a single bond, and the conformational strain barrier to that rotation is high enough to allow isolation of the conformers. The compounds of the present invention can be prepared as individual isomers by specific synthesis for any of the isomers, or can be resolved from a mixture of isomers. Conventional resolution techniques include using an optically active acid to form salts of the free bases of each isomer of an isomer pair (subsequently followed by fractional crystallization and regeneration of the free base), using an optically active amine to form salts of the acid forms of each isomer of an isomer pair (subsequently followed by fractional crystallization and regeneration of the free acid), using an optically pure acid, amine or alcohol to form esters or amides of each of the isomers of an isomer pair (subsequently followed by separation by chromatography and removal of the chiral auxiliary), or using various well-known chromatographic methods to resolve any isomer mixture of the starting material or the final product.

[0022] For example, if a particular enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or by induction using a chiral auxiliary, where the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to obtain the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group such as an amino group or an acidic functional group such as a carboxyl group, diastereomeric salts are formed using an appropriate optically active acid or base, and subsequently the diastereomers so formed are resolved by fractional crystallization or chromatographic means well known in the art, and then the pure enantiomer is recovered.

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

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

[0025] The structures illustrated in this specification also mean that they include compounds that differ only in that one or more isotope-enriched atoms are present. For example, replacement of hydrogen by deuterium or tritium, or 13 replacement of carbon by 14 C-enriched carbon to generate a compound is within the scope of the present invention.

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

[0027] As used herein, the terms "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refer to pharmaceutically acceptable materials, compositions or vehicles such as liquid or solid fillers, diluents, excipients, solvents or encapsulating materials that are involved in transporting or delivering the subject chemical substance 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 nonpyrogenic. 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) celluloses and their derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. In certain embodiments, the pharmaceutical compositions of the invention are nonpyrogenic, i.e., they do not induce a significant increase in temperature when administered to a patient.

[0028] The term "pharmaceutically acceptable salts" refers to addition salts of relatively non-toxic inorganic and organic acids of the compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified 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, among others. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19.)

[0029] In other cases, the compounds useful in the methods of the present invention may contain one or more acidic functional groups, thereby enabling the formation of pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term "pharmaceutically acceptable salts" in these cases refers to addition salts of relatively non-toxic inorganic and organic bases of the compound(s). These salts can likewise 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 the 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 salts, sodium salts, potassium salts, calcium salts, magnesium salts and aluminum salts, among others. Representative organic amines useful in the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc. (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 small molecules.

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

[0032] The term "preventive or therapeutic" treatment is recognized in the art and includes administration of one or more of the subject compositions to a host. When carried out prior to the clinical manifestation of an undesirable state (e.g., a disease or other undesirable state of a host animal), the treatment is preventive (i.e., protecting the host from the occurrence of the undesirable state), whereas when carried out after the manifestation of an undesirable state, the treatment is therapeutic (i.e., intended to reduce, ameliorate, or stabilize an existing undesirable state 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 a human.

[0034] Aliphatic chains include the classes of alkyl, alkenyl, and alkynyl as defined below. Straight-chain aliphatic chains are limited to unbranched carbon chain moieties. As used herein, the term "aliphatic group" refers to straight-chain, branched-chain, or cyclic aliphatic hydrocarbon groups, including saturated and unsaturated aliphatic groups such as alkyl groups, alkenyl groups, 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-chain or branched-chain alkyl has 30 or fewer carbon atoms in its main chain (e.g., C1-C 30 , and in the case of a branched chain, C3-C 30 ), more preferably, it has 20 or fewer carbon atoms. An 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 carbon atoms.

[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 that is substituted with at least one hydroxyl.

[0039] As used herein, the term "alkylene" refers to an alkyl group having the specified number of carbons, e.g., 2 to 12 carbon atoms, and containing two attachment points to the remainder of the compound on its longest carbon chain. Non-limiting examples of alkylene groups include methylene-(CH2)-, ethylene-(CH2CH2)-, n-propylene-(CH2CH2CH2)-, isopropylene-(CH2CH(CH3))-, and the like. The alkylene group can be a cyclic or acyclic, branched or unbranched carbon chain moiety and may optionally be substituted with one or more substituents.

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

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

[0042] "Cycloheteroalkyl" refers to a cycloalkyl moiety as previously defined containing one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms. Preferred cycloheteroalkyls have 4 to 8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4 to 6 carbons and heteroatoms in the ring structure. The cycloheteroalkyl group can be substituted or unsubstituted.

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

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

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

[0046] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups of 3 to 12 members, where each atom of the ring is carbon (i.e., carbocyclic aryl) or one or more atoms are heteroatoms (i.e., heteroaryl). Preferably, the aryl group includes a 5- to 12-membered ring, more preferably a 6- to 10-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings where two or more carbons are common to two adjacent rings, and in this case, 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, etc. Heteroaryl groups include substituted or unsubstituted aromatic ring structures of 3 to 12 members, more preferably 5 to 12 members, more preferably 5 to 10 members, and the ring structure contains 1 to 4 heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.

[0047] As used herein, the term "halo", "halide", or "halogen" means halogen and includes, for example, but not limited to, both radioactive and non-radioactive forms of fluoro, chloro, bromo, iodo, etc. In a preferred embodiment, halo is selected from the group consisting of fluoro, chloro, and bromo.

[0048] The term "heterocyclyl" or "heterocyclic group" refers to a ring structure having 3 to 12 members, more preferably a 5- to 12-membered ring, still more preferably a 5- to 10-membered ring, the ring structure containing 1 to 4 heteroatoms. The heterocycle can be monocyclic, bicyclic, spirocyclic, or polycyclic. Examples of the heterocyclyl group include thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolidine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, phthalan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactone, lactam, for example, azetidinone and pyrrolidinone, sultam, sultone, and the like. The heterocyclic ring can be substituted at one or more positions with substituents such as those described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN, and the like.

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

[0050] As used herein, the definition of each expression, such as alkyl, m, n, etc., is intended to be independent of its definition elsewhere in the same structure if it appears multiple times in any structure.

[0051] As used herein, "small molecule" refers to an organic or inorganic small molecule having a molecular weight of less than about 3,000 Daltons. Generally, the small molecules useful in the present invention have a molecular weight of less than 3,000 Daltons (Da). Small molecules can be, for example, at least about 100 Da to about 3,000 Da (e.g., about 100 to about 3,000 Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1500, about 500 to about 1000, about 300 to about 1000 Da, or about 100 to about 250 Da).

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

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

[0054] The terms "reducing," "decreasing," "decreased," "decrease," "reduction," and "inhibiting" are all used herein generally to mean a statistically significant decrease as compared to a reference. However, to avoid misunderstanding, "reducing," "decreasing," or "reducing" or "inhibiting" typically means at least a 10% decrease as compared to a reference concentration, for example, 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% decrease as compared to a reference concentration, including, for example, a complete absence, or any decrease between 10-99% as compared to the absence of a given treatment.

[0055] The terms "increased", "increasing", "enhanced", or "activated" are all used herein generally to mean an increase in a statistically significant amount, and to avoid misunderstanding, it is noted that the terms "increased", "increasing", "enhanced", or "activated" mean an increase of at least 10% compared to the 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 the 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 increase, or any increase between 2-fold - 10-fold or more compared to the reference concentration.

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

[0057] "Radiopharmaceutical", as defined herein, 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. Radio-labeled pharmaceuticals, e.g., radio-labeled antibodies, contain a radioisotope (RI) that functions as a radiation source. As contemplated herein, the term "radioisotope" includes both metallic and non-metallic radioisotopes. The radioisotope is selected based on the medical use of the radio-labeled 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 directly or via a linker to the remainder of the molecule.

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

[0059] The compounds of the present invention One aspect of the present invention is a compound of formula (I): [Chemical formula] (wherein, n is 0, 1, or 2, L1 is absent or is 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, provided 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 the presence of both of them together with the nitrogen atom to which they are attached forms a 5 - or 6 - membered heterocyclyl, Y3, Y4, Y5, and Y6 are independently selected from -H, -OH, halide, alkyl, haloalkyl, and alkoxy, provided 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 [Chemical formula] (not selected from) or a pharmaceutically acceptable salt thereof.

[0060] In certain embodiments, the compound is also [Chemical formula] not selected from or a pharmaceutically acceptable salt thereof.

[0061] In certain embodiments, the compound has a structure selected from the following: [Chemical formula]

[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, [Chemical formula] 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 [Chemical formula] is, or X2 is -H and X1 is [Chemical formula] .

[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-. In other embodiments, L1 is [Chemical formula] . In other embodiments, L1 is [Chemical formula] selected from. In other embodiments, L1 is [Chemical formula] selected from.

[0068] In certain embodiments, the compound is selected from the following: [Chemical formula]

[0069] In certain embodiments, Y1 is unsubstituted aryl, such as unsubstituted phenyl and unsubstituted naphthyl.

[0070] In certain embodiments, Y1 is substituted aryl.

[0071] In certain embodiments, Y1 is [Chemical formula] and R1, R2, R3, R4, and R5 are independently selected from -H, halogen, -CN, 、 -CF3, -CHF2, -CF2CH3, -OCF3, -OCHF2, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl, provided 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, [Chemical formula] and 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 [Chemical formula] and 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, Y1 is [Chemical formula] selected from.

[0076] In certain embodiments, Y1 is selected from, and Y1 is [Chemical formula] selected from

[0077] In certain embodiments, Y1 is unsubstituted heteroaryl.

[0078] In certain embodiments, Y1 is [Chemical formula] selected from

[0079] In certain embodiments, Y1 is substituted heteroaryl.

[0080] In certain embodiments, Y1 is [Chemical formula] 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 the following: [Chemical formula]

[0085] In certain embodiments, the compound is selected from the following: [Chemical formula]

[0086] In certain embodiments, the compound is selected from the following: [Chemical formula]

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

[0088] In certain embodiments, Y2 is [Chemical formula] selected from

[0089] In certain embodiments, Y2 is [Chemical formula] is

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

[0091] In certain embodiments, Y2 is [Chemical formula] and R 10 、R 11 、and R 12 are -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 is independently selected from, provided that R 10 R 11 and R 12 at least one of 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

Chemical formula

[0093] In certain embodiments, R 10 R11 and R 12 is independently selected from -H, -F, -Cl, -Br, -CN, -CH3, -CH2CH 3、 -CF3, -CHF2, -CF2CH 3、 -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 each -H, and R 11 is -CN, -CF3, -CH3, -OCH3, -NH2, -NHCH3, -NHAc, -CO2H, -C(O)NH2, -C(O)NHCH3, -C(O)NHCH2CH3,

Chemical formula

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

Chemical formula

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

[0097] In a particular embodiment, Y2 is [Chemical formula] selected from, R 16 For each occurrence, is halogen, -CN, -NH2, -OCF3, -OCHF2, -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino cycloalkyl, aryl, heteroaryl, -C(O)NR 13 R 14 , -CO2R 15 independently selected from, R 13 R 14 and R 15 each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

[0098] In a particular embodiment, R 16 is -CN, -CH3, -CF3, -C(O)NH2, -CO2CH2CH3, and [Chemical formula] selected from.

[0099] In a particular embodiment, Y2 is [Chemical formula] selected from, R 17 R 18 R 19 R 20 and R 21Each entity is independently selected from -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 provided that at least one of R 17 R 18 R 19 R 20 and R 21 is not -H, R 13 R 14 and R 15 each entity is independently selected from -H, alkyl, aryl, and heteroaryl.

[0100] In certain embodiments, Y2 is

Chemical formula

[0101] In certain embodiments, R 17 , R 18 , R 19 , R 20 , and R 21 are independently selected from -H, -CN, -CH3, and -OCH3.

[0102] In certain embodiments, Y2 is

Chemical formula

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

Chemical formula

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

Chemical formula

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

Chemical formula

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

[0107] In certain embodiments, Y2 is

Chemical formula

[0108] In certain embodiments, Y2 is

Chemical formula

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

[0110] In certain embodiments, Y2 is

Chemical formula

[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

Chemical formula

[0116] In certain embodiments, Y2 is

Chemical formula

[0117] In certain embodiments, Y2 is

Chemical formula

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

[0119] In certain embodiments, Y2 is

Chemical formula

[0120] In certain embodiments, Y2 is

Chemical formula

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

[0122] In certain embodiments, Y2 is

Chemical formula

[0123] In certain embodiments, each occurrence of R 22 R 23 R 24 and each occurrence of R 25 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

Chemical formula

[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, both Y3 and Y4 are -H or -F. In other embodiments, Y3 is selected from -F, -CF3, -OH and -OCH3, and Y4 is -H. In other embodiments, Y4 is selected from -F, -CF3, -OH and -OCH3, and Y3 is -H.

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

[0132] In certain embodiments, both Y5 and Y6 are -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, both Y5 and Y6 are -H. In other embodiments, both Y5 and Y6 are -F.

[0134] In some embodiments, the compound is selected from Table 1 below:

Table 1

[0135] In some embodiments, the compound is selected from Table 2 below: [Table 2] TIFF2025098065000071.tif211167 TIFF2025098065000072.tif240167 TIFF2025098065000073.tif225167 TIFF2025098065000074.tif250167 TIFF2025098065000075.tif243167 TIFF2025098065000076.tif250163 TIFF2025098065000077.tif246167 TIFF2025098065000078.tif233167

[0136] In some embodiments, the compound is selected from Table 3 below: [Table 3] TIFF2025098065000080.tif243167 TIFF2025098065000081.tif237167 TIFF2025098065000082.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] TIFF2025098065000085.tif250162 TIFF2025098065000086.tif246167

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

[0140] Any compound of the present invention can also be radiolabeled for the preparation of radiopharmaceuticals.

[0141] Treatment methods 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 by modulating SLC6A19 transport.

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

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

[0146] In some embodiments, the compound reduces the phenylalanine concentration in the subject's body.

[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, the method comprising administering to the subject an effective amount of a compound of formula (I).

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

[0149] In some embodiments, the present invention relates to a method of treating or preventing isovaleric academia, methylmalonic academia, propionic academia, maple syrup urine disease, DNAJC12 deficiency, urea cycle disorder, or hyperammonemia in a subject in need thereof, the method 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 a subject.

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

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

[0154] In some embodiments, the compound decreases the amino acid concentration in the body of the subject.

[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: [Chemical formula] (I) (wherein, 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, provided 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, heteroalkyl, 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 Y2’’s 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, provided 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:

Chemical formula

[0158] In some embodiments of any one of the disclosed methods, the compound is selected from:

Chemical formula

[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 Composition, Route of Administration, and Administration 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, the pharmaceutical composition comprises a plurality of compounds of the present invention and a pharmaceutically acceptable carrier.

[0165] In certain embodiments, the pharmaceutical composition of the present invention further comprises at least one additional pharmaceutically active agent other than the compound of the present invention. The at least one additional pharmaceutically active agent can be an agent useful for the treatment of ischemic reperfusion injury.

[0166] The pharmaceutical composition of the present invention can be prepared by combining one or more compounds of the present invention with a pharmaceutically acceptable carrier and optionally one or more additional pharmaceutically active agents.

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

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

[0169] Generally, the daily oral dosage of the compound is about 0.01 milligram / kg / day to 1000 milligrams / kg / day for human subjects. An oral dosage in the range of 0.5 to 50 milligrams / kg with more than one administration per day is expected to produce a therapeutic effect. The dosage can be appropriately adjusted to achieve the desired local or systemic drug concentration depending on the mode of administration. For example, in intravenous administration, the dosage per day is expected to be one to several orders of magnitude smaller. If the response in the subject at such a dosage is insufficient, higher dosages (or effective higher dosages by another more local delivery route) can be employed to the extent that the patient's tolerance permits. Multiple administrations per day are contemplated to achieve an appropriate in vivo concentration of the compound.

[0170] For any compound described herein, a therapeutically effective amount can first be determined from animal models. Also, a therapeutically effective amount can be determined from human data for compounds tested in humans and human data for compounds known to exhibit similar pharmacological activities such as other relevant active agents. In parenteral administration, higher dosages may be required in some cases. The dosage applied can be adjusted according to the relative bioavailability and efficacy of the compound administered. Adjusting the dosage to achieve the maximum effect based on the methods described above and other methods well known in the art is well within the ability of one of ordinary skill in the art.

[0171] The formulations of the present invention can be administered in a pharmaceutically acceptable solution that can routinely contain salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic components at pharmaceutically acceptable concentrations.

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

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

[0174] In the case of oral administration, the compound can be easily formulated by combining the active compound(s) with a pharmaceutically acceptable carrier well known in the art. Such carriers enable the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, etc. for oral ingestion by the subject to be treated. Pharmaceutical preparations for oral use can be obtained with solid excipients, and optionally, the resulting mixture can be ground, and if desired, after adding suitable auxiliaries, the granule mixture can be processed to obtain tablets or dragee cores. Suitable excipients include, in particular, fillers such as sugars including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, and / or polyvinylpyrrolidone (PVP). Optionally, cross-linked polyvinylpyrrolidone, agar, or a disintegrant such as alginic acid or its salts, such as sodium alginate, may be added. Optionally, oral formulations may also be formulated with physiological saline or buffer solutions, such as EDTA for neutralizing acidic conditions in the body, or may be administered without any carrier.

[0175] Also specifically contemplated are oral dosage forms of one or more of the above components. The one or more components can be chemically modified such that oral delivery of its derivative is effective. Generally, the contemplated chemical modification is to attach at least one moiety to the component molecule itself, and by this moiety, (a) acid hydrolysis is inhibited and (b) uptake from the stomach or intestine into the bloodstream becomes possible. Also, it is desirable to increase the overall stability of the one or more components and lengthen the circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, and polyproline. Abuchowski and Davis, “Soluble Polymer-Enzyme Adducts”, In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, N.Y., pp. 367-383 (1981); Newmark et al., J Appl Biochem 4:185-9 (1982). Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-thioxocane. For pharmaceutical use, as shown above, polyethylene glycol moieties are preferred.

[0176] The site of release of the component (or derivative) can be the stomach, small intestine (duodenum, jejunum, or ileum), or large intestine. One of ordinary skill in the art can obtain formulations that do not dissolve in the stomach but release the substance in the duodenum or other locations in the intestine. Preferably, the release avoids adverse effects in the gastric environment, either by protection of the compounds (or derivatives) of the present invention or by release of the biologically active substance, such as in the intestine after passing through the gastric environment.

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

[0178] Coating agents or mixtures of coating agents can also be used for tablets that are not intended to be protected from the stomach. This can include sugar coating agents or coating agents that make the tablets easier to swallow. Capsules can be composed of a hard shell (such as gelatin) for the delivery of dry therapeutic agents (e.g., powders), and in the case of liquid forms, a soft gelatin shell can be used. The shell material of cachets can also be thick starch or other edible paper. In the case of pills, lozenges, molded tablets, or powder tablets, wet granulation techniques can be used.

[0179] The therapeutic agent can also be included in the formulation as fine multi-particles in the form of granules or pellets with a particle size of about 1 mm. The formulation of the material for capsule administration can also be in the form of a powder, a lightly compressed plug, or a tablet. The therapeutic agent can be prepared by compression.

[0180] Both coloring agents and flavoring agents can be included. For example, the compounds (or derivatives) of the present invention can be formulated (such as encapsulation by liposomes or microspheres), and then further included in edible products such as refrigerated beverages containing coloring agents and flavoring agents.

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

[0182] A disintegrant may be included in the formulation of the therapeutic agent to form a solid dosage form. Materials used as disintegrants include, but are not limited to, starch, and there is a commercially available starch-based disintegrant, Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethyl cellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, carrageenan, and bentonite can all be used. Another form of disintegrant is an insoluble cationic exchange resin. Powdered gums can be used as binders and can include powdered gums such as agar, karaya, or tragacanth. Alginate and its sodium salts are also useful as disintegrants.

[0183] Binders can be used to hold the therapeutic agent and form hard tablets, 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 an alcohol solution to granulate the therapeutic agent.

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

[0185] A flow promoter that can improve the fluidity of the drug during formulation and assist in rearrangement during compression may be added. Flow promoters can include starch, talc, pyrogenic silica, and hydrated calcium aluminosilicate.

[0186] To assist in the dissolution of the therapeutic agent in an aqueous environment, a surfactant may be added as a wetting agent. Anionic surfactants such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate, and dioctyl sodium sulfonate may be included in the surfactant. Cationic surfactants may be used, and benzalkonium chloride and benzethonium chloride may be included. Potential non-ionic surfactants that can be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glyceryl monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid ester, methylcellulose, and carboxymethylcellulose. These surfactants can be present in the formulation of the compounds or derivatives of the present invention, either alone or as a mixture in different ratios.

[0187] Pharmaceutical preparations for oral use may include not only push-fit capsules made of gelatin, but also sealed capsules made of gelatin and plasticizers such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient in admixture with fillers such as lactose, binders such as starch, and / or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All formulations for oral administration must be in dosage amounts suitable for such administration.

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

[0189] For topical administration, the compound may be formulated as a solution, gel, ointment, cream, suspension, etc., as is well known in the art. Systemic formulations include those designed for administration by injection, for example, subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, and those designed for transdermal, transmucosal, oral or pulmonary administration.

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

[0191] Also contemplated herein is pulmonary delivery of the compounds (or salts thereof) disclosed herein. The compounds are delivered to the lungs of a mammal during inhalation and pass through the epithelial lining of the lungs to 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) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl.5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (a-1-proteinase); Oswein et al., 1990, “Aerosolization of Proteins”, Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-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 to Wong et al. issued September 19, 1995 (incorporated by reference).

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

[0193] Some specific examples of commercially available devices suitable for the practice of the present invention are the Ultravent nebulizer, manufactured by Mallinckrodt, Inc., St. Louis, Mo.; the Acorn II nebulizer, manufactured by Marquest Medical Products, Englewood, Colo.; the Ventolin metered dose inhaler, manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and the Spinhaler powder inhaler, manufactured by Fisons Corp., Bedford, Mass.

[0194] All such devices require the use of a formulation suitable for dispensing the compounds of the present invention. Typically, each formulation is specific to the type of device employed and may involve the use of appropriate propellant substances in addition to the normal diluents, adjuvants and / or carriers useful in therapy. Also contemplated is the use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers. Chemically modified compounds of the present invention can be formulated 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 the compound (or derivative) of the present invention in which the biologically active compound of the present invention is dissolved in water at a concentration of about 0.1 to 25 mg per mL of solution. The formulation may also contain buffers and simple sugars (e.g., for stabilization of the inhibitor and control of osmotic pressure). Nebulizer formulations may also contain surfactants to reduce or prevent aggregation induced on the surface of the compounds of the present invention which occurs upon atomization of the solution in forming the aerosol.

[0196] The formulations used with metered-dose inhalers generally contain micronized powders containing the compound (or derivative) of the invention suspended in a propellant, with the aid of a surfactant. The propellant can be any conventional material employed for this purpose, such as chlorofluorocarbons, hydrochlorofluorocarbons, hydrofluorocarbons, or hydrocarbons, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or combinations thereof. Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid can also be useful as a surfactant.

[0197] Formulations for dispensing from a powder inhaler include micronized dry powders containing the compound (or derivative) of the invention and may include bulking agents such as lactose, sorbitol, sucrose, or mannitol in an amount that facilitates dispersion of the powder from the device, for example, from 50 to 90% by weight of the formulation. The compound (or derivative) of the invention should preferably be prepared in particulate form having an average particle size of less than 10 micrometers (μm), most preferably from 0.5 to 5 μm, for the most effective delivery to the deep lung.

[0198] Transnasal delivery of the pharmaceutical compositions of the invention is also contemplated. Transnasal delivery allows the pharmaceutical compositions of the invention to enter directly into the bloodstream without the product depositing in the lungs after administration of the therapeutic product to the nose. Formulations for transnasal delivery include those with dextran or cyclodextran.

[0199] For transnasal administration, a useful device is a small rigid bottle fitted with a metered sprayer. In one embodiment, dosing is delivered by drawing the pharmaceutical composition solution of the invention into a chamber of a determined volume, the chamber having an aperture sized to aerosolize the aerosol formulation by forming a spray when the liquid in the chamber is compressed. The chamber is compressed to administer the pharmaceutical composition of the invention. In a specific embodiment, the chamber is in a piston configuration. Such devices are commercially available.

[0200] Alternatively, it is a plastic squeeze bottle having an opening or openings sized to aerosolize an aerosol formulation by forming a spray when squeezed. The opening is usually at the top of the bottle, which is generally tapered to fit partially into the passage of the nose for efficient administration of the aerosol formulation. Preferably, the nasal inhaler provides a measured amount of aerosol formulation for administering a measured amount of drug.

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

[0202] Pharmaceutical formulations for parenteral administration include an aqueous solution of the active compound in a water-soluble form. Further, a suspension of the active compound can be prepared as a suitable oily injection suspension. 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 to enable the preparation of highly concentrated solutions.

[0203] Alternatively, the active compound can be in powder form for reconstitution prior to use with a suitable vehicle, for example, sterile pyrogen-free water.

[0204] The compound can also be formulated into rectal or vaginal compositions such as suppositories or retention enemas 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 formulations can be formulated using suitable polymers or hydrophobic substances (e.g., as emulsions in acceptable oils) or ion exchange resins, or as poorly soluble derivatives, e.g., as poorly soluble salts.

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

[0207] Suitable liquid or solid pharmaceutical preparation forms are, for example, aqueous or saline solutions for inhalation, microencapsulation, scroll encapsulation, coating on fine gold particles, encapsulation in liposomes, atomization, aerosols, pellets for implantation in the skin, or those dried on sharp objects for rubbing on the skin. The pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or sustained release preparations of the active compound, and in their preparation excipients and additives and / or auxiliaries, such as disintegrants, binders, coating agents, swelling agents, lubricants, flavoring agents, sweeteners, or solubilizers, are customarily used as described above. The 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 optionally other therapeutic agents can be administered straight or in the form of a pharmaceutically acceptable salt or co-crystal. When used in medicine, the salt or co-crystal must be pharmaceutically acceptable, but pharmaceutically unacceptable salts or co-crystals can be used conveniently to prepare their pharmaceutically acceptable salts or co-crystals. 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. Also, such salts can be prepared as alkali metal salts or alkaline earth metal salts such as sodium salts, potassium salts or calcium salts of carboxylic acid groups.

[0209] Suitable buffers 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 composition of the present invention contains an effective amount of the compound described herein and optionally a therapeutic agent contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers, diluents or encapsulating substances suitable for administration to humans or other vertebrates. The term "carrier" refers to natural or synthetic organic or inorganic components combined with the active ingredient to facilitate application. The components of the pharmaceutical composition can also be mixed with the compounds of the present invention or with each other in such a way that there is no interaction that substantially impairs the desired pharmaceutical effect.

[0211] Although not particularly limited, the therapeutic agent(s) containing the compound of the present invention may be provided in the form of particles. As used herein, the particles mean nanoparticles or microparticles (or in some cases, larger particles) that may constitute all or part of the compound 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 particles. The therapeutic agent(s) may also be adsorbed onto the particles. The particles may have a release rate in any order, 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 of the substances conventionally used in the technical fields of pharmacy and medicine, including, but not limited to, disintegrating, non-disintegrating, biodegradable, or non-biodegradable substances or combinations thereof. The particles may be microcapsules containing the compound of the present invention in a solution or semi-solid state. The particles may be substantially in any shape.

[0212] For the production of particles for delivering the therapeutic agent(s), both non-biodegradable and biodegradable polymer materials can be used. Such polymers can be natural or synthetic polymers. The polymer is selected based on the period during which release is desired. Particularly interesting bioadhesive polymers include the biodegradable hydrogels described in Sawhney H S et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein by reference. These include polyhyaluronic acid, casein, gelatin, gluten, polyanhydrides, polyacrylic acid, alginates, 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 mode and profile of drug release from the formulation are controlled. This refers to immediate release and non-immediate release formulations, and non-immediate release formulations include, but are not limited to, sustained release and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in its conventional meaning and refers to a drug formulation that results in a slow release of the drug over a long period of time, preferably, although not necessarily, resulting in a substantially constant drug blood concentration over a long period of time. The term "delayed release" is used in its conventional meaning and refers to a drug formulation in which there is a time lag between the administration of the formulation and the release of the drug from the formulation. "Delayed release" may or may not be accompanied by a slow release of the drug over a long period of time, and thus 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 configured and arranged to deliver a therapeutic concentration of the active ingredient for at least 7 days, 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] Other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the description of the invention contained herein, taking into account the information known to those skilled in the art, and can be made without departing from the scope of the invention or any of its embodiments, as will be understood by those skilled in the relevant technical fields. Although the invention has been described in detail heretofore, the invention will be more clearly understood by reference to the following examples. The examples are included herein for illustrative purposes only and are not intended to limit the invention.

Examples

[0216] The present invention is further described in the following examples, but does not limit the scope of the present invention described in the claims.

[0217] Example 1: SLC6A19 Isoleucine Transport Assay Generation and Maintenance of Cell Lines The Flp-In™ T-REx™ 293 cell line was purchased from Thermo Fisher Scientific. Using this line, stable cell lines were generated that inducibly express human SLC6A19 containing a V5 tag at the C-terminus and stably express human TMEM27 (also known as Collectrin) containing a myc-DDK tag at the C-terminus. The stable cell lines were created 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 96-well cell culture-treated plates coated with poly-D-lysine. On day 1, the expression of SLC6A19 was induced by dispensing tetracycline at a final concentration of 1 μg / mL using a Tecan D300e digital dispenser. On day 2, a transport assay was performed. 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 175 μL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, the 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 incubating with the isoleucine substrate for 20 minutes at room temperature, the cells were washed with 175 μL of live cell imaging solution using the Blue Washer. The cells were then lysed with 150 μL of 15 μM D-Leucine-d10 in ultrapure water (CDN Isotopes). To facilitate lysis, the plates were shaken on a shaker at 700 rpm for at least 40 minutes. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the lysates of untreated cells. To ensure proper mixing of the standard curve, the plates were returned to the shaker for at least 2 minutes. The plates were then centrifuged at 4,000 rpm for 5 minutes to pellet cell debris and precipitates. The supernatant was diluted 1:10 with acetonitrile + 0.1% formic acid in a polypropylene plate.

[0219] Assay: Isoleucine transport assay in 384-well format On day 0, a stable cell line was seeded at a density of 20,000 cells / well into a poly-D-lysine-coated 384-well cell culture-treated plate containing a medium with 1 μg / mL of tetracycline using a Viaflo 384-well pipette. The transport assay was performed the next day (day 1). The medium was removed from the plate using the GentleSpin setting of a Centrifugal Blue Washer (Blue Cat Bio), and the cells were washed with 80 μL of live cell imaging solution (Thermo Fisher) using the Blue Washer. After washing, the cells were treated with 20 μL of either 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 the Blue Washer. The 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 on a shaker at 700 rpm for at least 2 hours. After lysis, a standard dilution curve of 13C6,15N-L-isoleucine was added to the wells containing the lysate of untreated cells. To ensure proper mixing of the standard curve, the plate was returned to the shaker for at least 5 minutes. The plate was then centrifuged at 4,000 rpm for 10 minutes to pellet cell debris and precipitates. The supernatant was diluted 1:10 with acetonitrile + 0.1% formic acid in a polypropylene plate.

[0220] 13C6,15N-L-isoleucine analysis was performed using RapidFire365-QTOF 6545 (Agilent). For quantitative sample analysis, automatic solid-phase extraction (HILIC H6 cartridge) was utilized followed by mass spectrometry injection. Samples were loaded using 95% acetonitrile and 0.1% formic acid, eluted directly from the cartridge using 5% acetonitrile and 0.1% formic acid, and ESI-MS (electrospray ionization) analysis was carried out. Quantification of the analyte was performed using Agilent Masshunter Quant software from high-resolution full-scan data.

[0221] General procedure A: [Chemical formula] Step 1: Synthesis of compound A2 A solution of bis(trichloromethyl) carbonate (BTC) (0.5 equivalent) in toluene was added dropwise to a solution of compound A1 (1 equivalent) in toluene at 0 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 15 minutes, then heated to 130 °C and stirred for 2 hours under a N2 atmosphere. After cooling, the mixture was concentrated to dryness under reduced pressure to obtain crude compound A2, which was used directly in the next step without further purification.

[0222] Step 2: Synthesis of compound A4 A solution of compound A2 (1 equivalent) in anhydrous DCM was added dropwise to a mixture of compound A3 (1 equivalent) and TEA (3 equivalents) in anhydrous DCM at 0 °C under a N2 atmosphere. The resulting mixture was stirred at 0 °C for 1 hour under a N2 atmosphere. Then, the mixture was diluted with water and extracted twice with EtOAc. The combined organic layers were separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH) to obtain 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

Chem.

[0224] Step 2: Synthesis of M3 To a solution of M2 (12 g, 38.36 mmol) in MeOH (200 mL), Pd / C (4.0 g, 10%) was added. The resulting mixture was stirred at room temperature for 3 h under a hydrogen atmosphere. Then, the mixture was filtered and the filtrate was concentrated under reduced pressure to give M3 (10 g, 35.93 mmol, yield 93.63%) 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), TFA (60 mL) was added. The resulting mixture was stirred at room temperature for 4 h. 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 min to change the pH to 8 - 9. Then, the mixture was filtered and the filtrate was concentrated under reduced pressure to give M4 (6 g, 33.7 mmol, yield 93.7%) 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)3 (20.7 g, 97.62 mmol) was added to the above mixture. The resulting mixture was stirred for 3 hours. Then, the mixture was concentrated under reduced pressure. The residue was diluted with EtOAc and washed with aqueous NaHCO3 to pH = 8. The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluting 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) were added AcOH (2.19 g, 36.54 mmol) and NaBH3CN (538 mg, 8.526 mmol). The resulting mixture was stirred at 80 °C for 16 hours. 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 Na2SO4 and concentrated to dryness. The crude product was purified by column chromatography on silica (eluting 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 obtain crude M7 (274 mg, 0.825 mmol, 54.26%) as the 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 minutes, 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 minutes. Then, the mixture was concentrated to dryness under reduced pressure. The crude product was purified by column chromatography on silica gel (eluting with 5% MeOH / DCM) to obtain Example 2 (103 mg, 0.247 mmol, yield 30.0%) as a pale yellow solid. LC / MS (ESI) m / z: 418 (M+H) + . 1 1H 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 19F NMR (400 MHz, DMSO-d6) δ -121.05 (s).

[0230] The compounds in the following table were prepared from the above-mentioned or commercially available suitable starting materials using the above general procedure A and Intermediate M7 of Example 2. [Table 6] TIFF2025098065000093.tif40169

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

[0232] [Chemical formula] 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 overnight to 150 °C. Then, the reaction mixture was 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 obtain a crude product, which was purified by flash column chromatography (ethyl acetate:heptane, 0:100~40:60) to give tert-butyl = N-methyl-N-[(3R)-1-pyridazin-3-yl-3-piperidyl]carbamate of A2 (1.57 g, 5.37 mmol, yield 53.70%). 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 hours. Then, the reaction mixture was concentrated under reduced pressure by rotary evaporation. The crude residue was dissolved in ethyl acetate, washed with 1 M NaOH and brine, and concentrated to give (R)-N-methyl-1-(pyridazin-3-yl)piperidine-3-amine of A3 (450 mg, 2.34 mmol, yield 43.59%), which was used without further purification.

Table 7

[0233] Example 128: Synthesis of 3-(4-chloro-3-methylbenzyl)-1-cyclopropyl-1-((R)-1-((S)-tetrahydrofuran-2-carbonyl)piperidin-3-yl)urea [Chemical formula] To a mixture of M2 (116 mg, 1.0 mmol) and HATU (474 mg, 1.29 mmol) in DCM (20 mL), TEA (253 mg, 2.5 mmol) was added dropwise at 0 °C. After stirring at room temperature for 30 minutes, M1 (200 mg, 0.83 mmol) was added to the above mixture. The resulting mixture was stirred at room temperature for 2 hours under a N2 atmosphere. Then, the mixture was 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 obtain 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), TFA (2 mL) was added dropwise at 0 °C. The resulting mixture was stirred at room temperature for 2 hours. Then, the reaction mixture was evaporated to dryness under reduced pressure to obtain the crude product 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 hour. Then, NaBH(OAc)3 (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, filtered, and the filtrate was evaporated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel (eluting 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 NaBH3CN (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 solution (20 mL) and extracted twice with EtOAc (30 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluting 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 for 3 hours under a N2 atmosphere. 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] A solution of M10 (22.9 mg, 0.13 mmol) in anhydrous DCM (2 mL) was added dropwise to a mixture of M9 (30 mg, 0.13 mmol) and TEA (39 mg, 0.39 mmol) in anhydrous DCM (10 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 1 h under a N2 atmosphere. The reaction mixture was then diluted with water (20 mL) and extracted twice with DCM (20 mL). The combined organic layers were separated, dried over anhydrous Na2SO4, 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, H2O / 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 prepared by the same route as Example 128, starting from the appropriate commercially available amide M2 and other intermediates described above or commercially available.

Table 8

[0236] The following examples were synthesized according to general procedure A using commercially available basic components.

Table 9

[0237] Synthesis of general intermediates, Methods B - G

Chemical formula

[0238] General procedure B:

Chemical formula

[0239] Synthesis example of general procedure B:

Chemical formula

[0240] General Procedure C:

Chemical Structure

[0241] Synthesis Example of General Procedure C:

Chemical Structure

[0242] General procedure D:

Chem.

[0243] Synthesis example of general procedure D:

Chem.

[0244] Example of general procedure E:

Chemical formula

[0245] Example of general procedure F:

Chemical formula

[0246] Example of General Procedure G:

Chemical Structure

[0247] The examples in the following table were prepared from the aforementioned appropriate starting materials or commercially available starting materials using a method similar to the above.

Table 10

[0248] General procedure H: As a general procedure, the following examples were synthesized according to the following general scheme.

Chemical formula

[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 [Chemical formula] To a solution of M1 (200 mg, 0.952 mmol) in toluene (10 mL) were added TEA (192 mg, 1.904 mmol) and DPPA (314 mg, 1.142 mmol). The resulting mixture was stirred at 110 °C for 2 h under a N2 atmosphere. After cooling, the mixture was concentrated under reduced pressure to give crude M2 (197 mg, yield 99.9%) without further purification. To a solution of M3 (207 mg, 0.952 mmol) in dehydrated DCM (10 mL) were added TEA (288 mg, 2.856 mmol) and M2 (197 mg, 0.952 mmol) at 0 °C. The resulting mixture was stirred at room temperature for 30 min under a N2 atmosphere. Then, the mixture was 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, yield 15.1%) 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 from the above-described or commercially available appropriate starting materials using the above general procedure B detailed in Example 184.

Table 11

[0251] General procedure I: As a general procedure, the following examples were synthesized according to the following general scheme.

Chemical formula

[0252] Example 232: Synthesis of (R)-3-(4-chloro-3-methylbenzyl)-1-methyl-1-(1-(pyrimidine-4-carbonyl)piperidin-3-yl)urea

Chemical formula

[0253] The compounds in the following table were prepared from the above-mentioned or commercially available suitable starting materials using the general procedure I above and intermediate M1 of Example 232.

Table 12

[0254] General procedure J: Example 272: Synthesis of (R)-3-(4-chloro-3-methylbenzyl)-1-(1-(cyclopropanecarbonyl)piperidin-3-yl)-1-cyclopropylurea

Chemical formula

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

[0256] General Procedure K: Example 274: Synthesis of (R)-3-(4-chloro-3-methylbenzyl)-1-methyl-1-(1-(6-(trifluoromethyl)pyridazin-3-yl)piperidin-3-yl)urea [Chemical Structure] 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, yield 9.1%) 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

[0258] General Procedure M:

Chemical Structure

[0259] Step 2 M2 (1 equivalent) was dissolved in 20% piperidine / DMF (0.1 M), and the reaction mixture was stirred at room temperature for 1 - 2 hours. Subsequently, the reaction mixture was concentrated under vacuum, and the crude product was purified by flash column chromatography using DCM / DCM:7N NH4 MeOH (10:0 - 0:10) to obtain product M3.

[0260] Step 3 M3 (1 equivalent), the appropriate carboxylic acid M4 (1.2 equivalents), and HATU (1.3 equivalents) were dissolved in DMF (0.1 M), then DIEA (2 equivalents) 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 obtain 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 [Chemical formula] 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 dropwise with a solution of 5-(isocyanatomethyl)-3-phenylisoxazole (220 mg, 1.10 mmol) in anhydrous DCM (5.5 mL) at 0 °C under a N2 atmosphere. The resulting mixture was stirred at 0 °C for 1 h under a N2 atmosphere. The mixture was then diluted with water and extracted twice with EtOAc. The combined organic layers were 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 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 under vacuum, 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), then DIEA (21 uL, 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 under vacuum. 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 the general procedure M.

Table 16

[0263] General procedure N [Chemical formula] A solution of compound (R)-N-cyclopropyl-1-((2-nitrophenyl)sulfonyl)piperidin-3-amine (1 equiv) and TEA (3 equiv) in anhydrous DCM was added dropwise with a solution of compound N1 (1 equiv) in anhydrous DCM at 0 °C under an N2 atmosphere. The resulting mixture was stirred at 0 °C for 1 h under an N2 atmosphere. Then, the mixture was diluted with water and extracted twice with EtOAc. The combined organic layers were separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (eluting with heptane / ethyl acetate) to give compound N2. N2 (1 equiv) was dissolved in DMF (0.1 M), potassium carbonate (2 equiv) and thiophenol (1.5 equiv) were added, and the reaction mixture was stirred at room temperature for 1 - 2 h. 1 M HCl was added until pH 2 - 3 was reached, and the resulting mixture was extracted twice with DCM. Then, the acidic solution was adjusted to pH 10 - 11 by carefully adding 6 N NaOH. Then, the resulting basic solution was extracted with DCM (x3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product 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 procedure O:

Chemical formula

[0265] Synthesis of O3 Appropriate amine O2 (2 equivalents) and CDI (2 equivalents) were dissolved in DMF (0.1 M), and the reaction mixture was stirred at room temperature for 2 - 3 hours. Then, O1 (1 equivalent) was added, and the reaction mixture was stirred at 50 °C until complete conversion (1 - 4 hours). The reaction mixture was concentrated under vacuum, and the crude product was purified by preparative HPLC to obtain the product O3.

[0266] (3R)-N-Cyclopropyl-3-(3-cyclopropyl{[(4-chloro-2-fluoro-5-methylphenyl)methyl]carbamoyl}amino)piperidine-1-carboxamide synthesis (Example 317)

Chemical formula

Table 17

[0267] General procedure P:

Chem.

[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 hours in an ice bath. The reaction mixture was then stirred for an additional 1 hour, 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 dioxane / HCl (4.0 M) (4.0 M, 59.17 mL), and the reaction mixture was stirred at 60 °C for 3 hours. The reaction mixture was then concentrated under reduced pressure by rotary evaporation, and the resulting solid was washed twice with Et2O 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) were 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 and 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.A solution of 9H-fluoren-9-ylmethyl (3R)-3-[(2,4-dimethoxyphenyl)methylamino]piperidine-1-carboxylate (5.75 g, 12.17 mmol) in THF (50 mL) and ethanol (100 mL) was added with (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). 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 solution and brine, and dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by chromatography (DCM:MeOH 10:0~9:1) to obtain 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 hours. Then, the reaction mixture was cooled to room temperature and concentrated. The residue was dissolved in DCM, washed with saturated Na2CO3 aqueous solution, dried over anhydrous Na2SO4, and concentrated to obtain 9H-fluoren-9-ylmethyl (3R)-3-(cyclopropylamino)piperidine-1-carboxylate P6 (1.35 g, 3.72 mmol, yield 95.47%) as a brown oil, which was stored at -20 °C and used without further purification.

[0269] General procedure Q: [Chemical formula] 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), 2-nitrobenzenesulfonyl chloride (11.62 g, 52.43 mmol) was added portionwise as a solid. The reaction mixture was stirred at ambient temperature for 15 minutes, then TFA (56.93 g, 499.31 mmol, 38.47 mL) was added slowly. The reaction mixture was stirred at ambient temperature for a further 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 were 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 at ambient temperature overnight. The reaction mixture was then washed with 1 M NaOH (500 mL), then the organic layer was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure by rotary evaporation. The crude residue was purified by flash column chromatography (DCM:MeOH, 100:0~96:4) 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) were added 1-ethoxy-1-trimethylsilyloxycyclopropane (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.Next, the residue was dissolved in ethyl acetate (250 mL), washed with 1 M NaOH (250 mL) and brine (250 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure by rotary evaporation to obtain (3R)-N-cyclopropyl-N-[(2,4-dimethoxyphenyl)methyl]-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (8.58 g, 18.04 mmol, yield 99.08%) 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 hours, an additional portion of triethylsilane (7.28 g, 62.61 mmol, 10 mL) was added to suppress the formation of 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 (ca. 200 mL) and brine (ca. 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 slowly added dropwise. The product was filtered from the solution to obtain (3R)-N-cyclopropyl-1-(2-nitrophenyl)sulfonyl-piperidin-3-amine (5.5 g, 15.20 mmol, yield 84.25%, HCl) Q4 as a tan solid.

[0270] General procedure R Intermediates of benzylamine bonded to a heterocycle on an aromatic ring were prepared by a general procedure.

Chemical formula

[0271] Synthesis of (2-fluoro-4-(oxazol-2-yl)phenyl)methanamine R6 according to general procedure R

Chemical formula

[0272] Examples 323 to 333 were prepared from the corresponding aryl or heteroaryl-benzylamine and the general procedure described above.

Table 18

[0273] The general procedure R’ described below was used to prepare Examples 334 to 339 and 343 and similar compounds.

Chemical formula

[0274] The general procedure S described below was used to prepare Examples 340, 341, and 342 and 343 and similar compounds.

Chemical formula

[0275] Examples 334 to 343

Table 19

[0276] The 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 the general procedure R following the general procedure C.

[0278] Examples 166, 192, 193, 201, 206, and 212 were prepared by General Procedure R following General Procedure B.

[0279] Example 335 was prepared by General Procedure R following General Procedure C.

[0280] Example 200 was prepared by General Procedure R following General Procedure D.

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

[0282] Example 180 was prepared by General Procedure S following General Procedure C.

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

[0284] General Procedure S’

Chemical Formula

[0285] Step 2: Synthesis of S3 S2 (1 equivalent) was dissolved in a solution of DCM / TFA 5:1 (0.1 M), and the resulting mixture was stirred at room temperature for 2 hours. Subsequently, the mixture was concentrated under reduced pressure to obtain 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 equivalent) in anhydrous DCM (0.03 M), 2,4-dimethoxybenzaldehyde (1.2 equivalents) and acetic acid (2 equivalents) were added, and the resulting mixture was stirred at room temperature for 1 hour. Subsequently, NaBH(OAc)3 (2 equivalents) was added to the above mixture, and the resulting mixture was stirred at 45 °C overnight. Then, the mixture was poured into a 5% Na2CO3 solution and extracted with DCM (×4). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH) to obtain compound S4.

[0287] Step 4: Synthesis of S5 To a mixture of S4 (1 equivalent) and acetic acid (10 equivalents) in THF / EtOH 1:2 (0.03 M), (1-ethoxycyclopropoxy)trimethylsilane (1.1 equivalents) was added, followed by the addition of NaBH3CN (3 equivalents). The resulting mixture was stirred at 80 °C for 16 hours under a N2 atmosphere. Then, the mixture was poured into a 5% Na2CO3 solution and extracted with DCM (×3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluted with DCM / MeOH) to obtain compound S5.

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

[0289] Step 6: Synthesis of S8 To a solution of S6 (1 equiv) in anhydrous DCM (0.03 M), DIEA (3 equiv) was added. The resulting mixture was stirred at 0 °C for 15 minutes and then added to a mixture of the corresponding isocyanate S7 (1 equiv) in anhydrous DCM (0.08 M) at 0 °C under N2 atmosphere. The resulting mixture was stirred at 0 °C to room temperature for 30 minutes. Then, the mixture was poured into saturated NaHCO3 solution and extracted with DCM (×3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to obtain pure compound S8.

[0290] Synthesis of Example 346 [Chemical formula] 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), TMSNCO (119 mg, 1.032 mmol) was added dropwise at 0 °C under N2 atmosphere. The resulting mixture was stirred at room temperature for 16 hours. Then, the mixture was poured into H2O (50 mL) and washed twice with EtOAc (40 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with DCM:MeOH = 100:0 to 20:1) to obtain compound 346-2 (150 mg, yield 83.3%) 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), TFA (1 mL) was added. The resulting mixture was stirred at room temperature for 2 hours. Then, the mixture was concentrated under reduced pressure to obtain crude compound 346-3 (158 mg, yield 99.0%) 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) were added 2,4-dimethoxybenzaldehyde (115 mg, 0.693 mmol) and acetic acid (69 mg, 1.155 mmol). Then the mixture was stirred at room temperature for 1 hour, after which NaBH(OAc)3 (244 mg, 1.155 mmol) was added to the above mixture and the resulting mixture was stirred at 45 °C overnight. Next, the mixture was poured into 5% Na2CO3 solution (50 mL) and extracted with DCM (30 mL × 4). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with DCM / MeOH = 100:0 to 12:1) to give compound 346-4 (176 mg, yield 97.8%) 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) was added (1-ethoxycyclopropoxy)trimethylsilane (111 mg, 0.637 mmol), followed by the addition of NaBH3CN (109 mg, 1.736 mmol). The resulting mixture was stirred at 80 °C for 16 hours under a N2 atmosphere. Next, the mixture was poured into 5% Na2CO3 solution (50 mL) and extracted with DCM (30 mL × 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with DCM / MeOH = 100:0 to 20:1) to give compound 346-5 (193 mg, yield 97.5%) 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. Then, the mixture was concentrated to dryness under reduced pressure. The crude product was purified by flash column chromatography (eluting with DCM / MeOH = 100:0 to 15:1) to give compound 346-6 (78 mg, yield 70.9%) 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. Then, a mixture of 2-fluoro-1-(isocyanatomethyl)-4-(trifluoromethoxy)benzene (92 mg, 0.388 mmol) in anhydrous DCM (5 mL) was added thereto at 0 °C under a N2 atmosphere. The resulting mixture was stirred at 0 °C to room temperature for 30 min. Then, the mixture was poured into saturated NaHCO3 solution (50 mL) and extracted with DCM (30 mL×3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give pure 346 (27 mg, yield 16.0%) 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 T [Chem.] Step 1: Synthesis of T2 To a mixture of the corresponding amine T1 (1 equiv) in a 5:1 (0.4 M) mixture of MeCN / DMF, CDI (1 equiv) was added at room temperature, and the resulting mixture was stirred for 2 h. The mixture was then concentrated under reduced pressure to give the crude product 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), TEA (4 equiv) was added, 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 (eluting with DCM:MeOH) to give T4.

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

[0299] Synthesis of Example 347 [Chem.] 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), CDI (744 mg, 4.588 mmol) was added at room temperature, and the resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give the crude product 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 Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting 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 [Chem.] U1 was synthesized according to procedure P or Q by deprotecting intermediate P5 and Q3 respectively.

[0304] Step 1: Synthesis 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, yield 75.7%) as a pale yellow solid. LC / MS (ESI) m / z: 334 (M+H) + .

[0305] Step 2: Synthesis of U3 Compound U2 (10 g, 29.99 mmol) was added portionwise at 0 °C under a N2 atmosphere to TFA (100 mL), and the resulting mixture was 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, yield 99.8%) 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 dropwise a mixture of the appropriate isocyanate U4 (1 equiv) and TEA (10 equiv) in DCM (0.15 M) at 0 °C under a N2 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 Na2SO4, 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 [Chemical formula] To a solution of U3 (22.4 mg, 0.095 mmol) in anhydrous DCM (0.4 mL) was added dropwise 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) at 0 °C under a N2 atmosphere. 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 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 352 (15 mg, yield 37.7%) as a white solid. LC / MS (ESI) m / z: 419.2 (M+H) + . 11H 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: [Chemical formula] V3 was synthesized using procedure M or N.

[0309] Step 1: Synthesis of V2 To a solution of the appropriate V1 (1 equiv) in THF (0.3 M), TEA (3 equiv) and CDI (1.2 equiv) were added, 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 (eluting with PE:EtOAc) to give compound V2.

[0310] Step 2: Synthesis of 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. Then, the mixture was concentrated to dryness under reduced pressure, the residue was dissolved in EtOAc, and washed with water and brine. Then, the organic layer was 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 obtain compound V4.

[0311] Synthesis of Example 348

Chemical formula

[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) and TEA (12 mg, 0.12) 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 Na2SO4, filtered, and concentrated to dryness. The crude product was purified by column chromatography on silica gel (eluting with PE:EtOAc = 100:0 to 1:1) to give compound 348-3 (19.5 mg, yield 82.29%) 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 adjust the pH to 8-9, and then the organic layer was 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 (eluting with DCM:MeOH = 100:0 to 20:1) to give 348 (15 mg, yield 96.2%) 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 following table were prepared by applying the general procedures G, R', and S of the above experiments. [Table 21] TIFF2025098065000237.tif223167 TIFF2025098065000238.tif72169

[0316] Examples 378 to 400 listed in the following table were prepared by applying the general procedures W, X, or Y of the experiments listed after the table. [Table 22] TIFF2025098065000240.tif241170 TIFF2025098065000241.tif236170 TIFF2025098065000242.tif226170 TIFF2025098065000243.tif237170 TIFF2025098065000244.tif250163 TIFF2025098065000245.tif249166 TIFF2025098065000246.tif237170 TIFF2025098065000247.tif231170 TIFF2025098065000248.tif232170 TIFF2025098065000249.tif211170

[0317] General procedure W: [Chemical formula] Step 1: Synthesis of W2 To a mixture of the appropriate aldehyde W1 (1 equiv) and hydroxylamine hydrochloride (1.1 equiv) in DCM, TEA (1.1 equiv) was added dropwise at 0 °C under an N2 atmosphere. The resulting mixture was stirred at room temperature for 45 minutes. Then, the mixture was 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 obtain 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 equiv) and W3 (1 equiv) in MeOH / H2O (4:1, v / v), [bis(trifluoroacetoxy)iodo]benzene (1.3 equiv) was added at 0 °C under an N2 atmosphere. The resulting mixture was stirred at room temperature for 1 hour. Then, the mixture was diluted with water and extracted with EtOAc. The combined organic layers were separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with PE / EtOAc) to obtain compound W4.

[0319] Step 3: Synthesis of W5 W4 (1 equiv) 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. Then, the mixture was concentrated to dryness under reduced pressure to obtain 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 equiv) and NaHCO3 (3 equiv) in DCM, a solution of triphosgene (0.5 equiv) in DCM was added at -30 °C under an N2 atmosphere. The resulting mixture was stirred at room temperature for 30 minutes, then a mixture of W6 (1 equiv) and TEA (3 equiv) in anhydrous DCM was added dropwise at 0 °C under an N2 atmosphere. The resulting mixture was stirred at room temperature for 1 hour, then the mixture was diluted with water and extracted twice with DCM. The combined organic layers were separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to obtain pure compound W.

[0321] Synthesis of Example 400: [Chemical formula] 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), TEA (586 mg, 5.79 mmol) was added dropwise at 0 °C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 45 minutes. Then, the mixture was diluted with water (25 mL) and extracted twice with DCM (25 mL). The combined organic layers were separated, washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to obtain crude compound W2 (950 mg, yield 88.05%), 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 H2O (10 mL), [bis(trifluoroacetoxy)iodo]benzene (2.59 g, 6.02 mmol) was added at 0 °C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 1 hour. Then, the mixture was diluted with water (60 mL) and extracted twice with EtOAc (50 mL). The combined organic layers were separated, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography (eluting with 10 - 50% EtOAc / PE) to obtain pure compound W4 (390 mg, yield 23.50%) 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), TFA (1 mL) was added at 0 °C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 1 hour. Then, the mixture was concentrated to dryness under reduced pressure to obtain the 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 the crude product W5 (60 mg, 0.232 mmol) and NaHCO3 (59 mg, 0.697 mmol) in DCM (5 mL) at -30 °C under a N2 atmosphere, and the resulting mixture was stirred at room temperature for 30 minutes. 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 N2 atmosphere, and the resulting mixture was stirred at room temperature for 1 hour. Then, the mixture was diluted with water (10 mL) and extracted twice with DCM (10 mL). The combined organic layers were separated, washed with brine (12 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to obtain 400 (22 mg, yield 20.25%) as a white solid. LC / MS: m / z 468 (M+H) + . 11H 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:

Chemical formula

[0326] Step 2: Synthesis of 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 for 2 h under a N2 atmosphere. After cooling, the mixture was concentrated to dryness under reduced pressure, and the residue was purified by flash column chromatography (eluting with PE / EtOAc) to obtain compound X3.

[0327] Step 3: Synthesis of 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. Then, the mixture was quenched with water and extracted twice with EtOAc. The combined organic layers were 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 obtain compound X4.

[0328] Step 4: Synthesis of 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 for 20 hours under a N2 atmosphere. Then, the mixture was diluted with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to obtain the crude product X5, which was used in the next step without further purification.

[0329] Step 5: Synthesis of X6 PPh3 (2 equivalents) was added to a mixture of X5 (1 equivalent) in THF / H2O (4:1, V / V), and the resulting mixture was stirred at room temperature for 17 hours under a N2 atmosphere. Then, the mixture was diluted with 2N HCl (aqueous solution) and washed with MTBE. The aqueous layer was separated, adjusted to pH 8 with saturated aqueous NaHCO3, and then extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to obtain the crude product X6, which was used in the next step without further purification.

[0330] Step 6: Synthesis of X A solution of triphosgene (0.5 eq) in DCM was added dropwise to a mixture of X6 (1 eq) and NaHCO3 (3 eq) in anhydrous DCM at -30 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 30 minutes. Subsequently, a mixture of X7 (1 eq) and TEA (3 eq) 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. Then, the mixture was 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 obtain Compound X.

[0331] General procedure Y: [Chemical formula] Step 1: Synthesis of Y2 To a solution of Y1 (1 eq) in MeCN, t-BuONO (1.2 eq) and TMSN3 (1.2 eq) were added at 0 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 1 hour. When TLC indicated 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 A solution of Y2 (1 eq) in MeCN was added dropwise to a mixture of Y3 (1.2 eq), sodium ascorbate (0.2 eq) and CuSO4 (0.2 eq) in MeCN at 0 °C under a N2 atmosphere. The resulting mixture was stirred overnight at room temperature under a N2 atmosphere. Then, the mixture was 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 obtain 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. Subsequently, the mixture was concentrated to dryness. The residue was dissolved in EtOAc, neutralized with an aqueous NaHCO3 solution, and extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to obtain crude product 5, which was used directly in the next step without further purification.

[0334] Step 4: Synthesis of Y7 A solution of TEA (3.0 equivalents) and BTC (0.5 equivalent) in DCM was added dropwise to a mixture of Y5 (1.0 equivalent) and Y6 (1.0 equivalent) in DCM at -78 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 1 hour. Subsequently, the mixture was 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 obtain Y7.

[0335] Synthesis of Example 386:

Chemical formula

[0336] Step 2: Synthesis of Y4 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 dropwise with a mixture of Y2 in MeCN (30 mL) at 0 °C. The resulting mixture was stirred overnight at room temperature under a N2 atmosphere. Then the mixture was 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 (eluting with PE / EtOAc = 100:0 to 3:2) to obtain pure Y4 (2.3 g, yield 37.80%). LC / MS (ESI) m / z: 359 (M+H) + .

[0337] Step 3: Synthesis of Y5 TFA (2 mL) was added dropwise to a solution of Y4 (80 mg, 0.223 mmol) in DCM (6 mL) 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 to pH = 8 with aqueous NaHCO3. The mixture was extracted twice with EtOAc (10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated to give crude Y5 (55 mg, yield 96.49%), 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 A solution of TEA (64 mg, 0.636 mmol) and BTC (31 mg, 0.106 mmol) in DCM (1 mL) was added dropwise to a mixture of Y5 (55 mg, 0.212 mmol) and Y6 (46 mg, 0.212 mmol) in DCM (5 mL) at -78 °C under a N2 atmosphere. The resulting mixture was stirred at room temperature for 1 hour. Then, the mixture was 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 Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give 386 (15 mg, yield 9.74%) as a white solid. LC / MS (ESI) m / z: 500 (M+H) + . 1 1H NMR (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] The representative compounds selected from the above were tested in the isoleucine transport assay described in Example 1. The results are summarized in the tables of Figures 2 and 3.

[0340] Incorporation by reference All U.S. patents and U.S. patent application publications cited herein are hereby incorporated by reference into this specification.

[0341] Equivalents Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

1. Compounds of formula (I): 【Chemistry 1】 (In the formula, n is 0, 1, or 2; L １ is absent or is selected from -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH ２ - selected from L ２ is absent or -CH ２ - and L ３ is absent or is —C(O)—, X １ and X ２ is independently selected from -H, alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl, with the proviso that X １ and X ２ are not both -H, Y １ is selected from aryl and heteroaryl; Y ２ is an alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NH(Y ２ '), and -N(Y ２ '') ２ is selected from Y ２ ' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, and cycloalkyl; Each Y ２ " is alkyl, or both occurrences together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocyclyl; Y ３ , Y ４ , Y ５ , and Y ６ is independently selected from -H, -OH, halide, alkyl, haloalkyl, and alkoxy, with the proviso that Y ３ and Y ４ Or Y ５ and Y ６ are not both -OH, However, L ３ When is -C(O)-, Y ２ is not aryl and the compound is 【Chemistry 2】 (not selected from or a pharma- ceutically acceptable salt thereof.

2. Y ２ The compound of claim 1, wherein ' is selected from -H, -OH, alkyl, alkoxy, alkoxyalkyl, and cycloalkyl.

3. X １ and X ２ is -H, and X １ and X ２ The other is C １ -C ４ The compound of claim 1 or 2, wherein the alkyl group is selected from alkyl, haloalkyl, cycloalkyl, alkyl-cycloalkyl, and heterocyclyl.

4. X １ and X ２ is -H, and X １ and X ２ The other is -CH ３ , -CH ２ CH ３ , -CH ２ CF ３ , -CH ２ CH ２ CH ３ , 【Chemistry 3】 The compound of claim 3 selected from:

5. X １ is -H, and X ２ But -CH ３ 5. The compound of claim 4,

6. X ２ is -H, and X １ But -CH ３ 5. The compound of claim 4,

7. X １ is -H, and X ２ but, 【Chemistry 4】 5. The compound of claim 4,

8. X ２ is -H, and X １ but, 【Chemistry 5】 5. The compound of claim 4,

9. L １ The compound according to any one of claims 1 to 8, wherein is absent.

10. L １ is -alkyl-, -hydroxyalkyl-, -cycloalkyl-, and -heteroaryl-CH ２ The compound according to any one of claims 1 to 8, selected from:

11. L １ But -CH ２ -, -C(H)(CH ３ ) -, -CH ２ CH ２ - and -C(H)(OH)CH ２ The compound according to claim 10, wherein said compound is selected from the group consisting of:

12. L １ but, 【Chemistry 6】 11. The compound of claim 10,

13. L １ but, 【Chemistry 7】 13. The compound of claim 12, selected from:

14. L １ but, 【Chemistry 8】 11. The compound of claim 10, selected from:

15. The compound of any one of claims 1, 2, and 9-14, having a structure selected from: 【Chemistry 9】

16. Y １ The compound of any one of claims 1 to 15, wherein is unsubstituted aryl.

17. Y １ 17. The compound of claim 16, wherein is selected from unsubstituted phenyl and unsubstituted naphthyl.

18. Y １ The compound of any one of claims 1 to 15, wherein is substituted aryl.

19. Y １ but, 【Chemistry 10】 and R １ , R ２ , R ３ , R ４ , and R ５ is -H, halogen, -CN, 、 -CF ３ , -CHF ２ , -CF ２ CH ３ , -OCF ３ , -OCHF ２ , alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl, where R １ , R ２ , R ３ , R ４ , and R ５ The compound of claim 18, wherein one of is not -H.

20. R １ ,R ２ ,R ３ ,R ４ 、RRR ５ が、-H、-D、-C-、-Br、-CN、-CH ３ 、-CH ２ CH ３、 -CF ３ 、-CHF ２ 、-CF ２ CH ３、 -OCH ３ 、-OCF ３ ,-OCHF ２ 、 【Chemistry 11】 20. The compound of claim 19, independently selected from:

21. R １ , R ２ , R ３ , R ４ , and R ５ is -H, -F, -Cl, -Br, -CN, -CH ３ , -CH ２ CH ３、 - OCF ３ , and 【Chemistry 12】 20. The compound of claim 19, independently selected from:

22. R １ , R ２ , R ３ , R ４ , and R ５ The compound according to any one of claims 19 to 21, wherein two of are not -H.

23. R １ , R ２ , R ３ , R ４ , and R ５ The compound according to any one of claims 19 to 21, wherein three of are not -H.

24. Y １ but, 【Chemistry 13】 20. The compound of claim 19, selected from:

25. Y １ but, 【Chemistry 14】 20. The compound of claim 19, selected from:

26. Y １ The compound of any one of claims 1 to 15, wherein is unsubstituted heteroaryl.

27. Y １ but, 【Chemistry 15】 24. The compound of claim 23, selected from:

28. Y １ The compound of any one of claims 1 to 15, wherein is substituted heteroaryl.

29. Y １ but, 【Chemistry 16】 is selected from R ６ , R ７ , R ８ , and R ９ Each occurrence of is -H, halogen, -CN, -OCF ３ , -OCHF ２ , alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, aryl, and heteroaryl, where R ６ , R ７ , R ８ , and R ９ The compound of claim 28, wherein at least one of is not -H.

30. L ２ The compound according to any one of claims 1 to 29, wherein is absent.

31. L ２ But -CH ２ The compound according to any one of claims 1 to 29, wherein

32. L ３ The compound according to any one of claims 1 to 31, wherein is absent.

33. L ３ The compound according to any one of claims 1 to 31, wherein is -C(O)-.

34. The compound according to any one of claims 1 to 33, wherein n is 0.

35. The compound according to any one of claims 1 to 33, wherein n is 1.

36. The compound according to any one of claims 1 to 33, wherein n is 2.

37. 35. The compound of any one of claims 1, 32, and 34, having a structure selected from: 【Chemistry 17】

38. 36. The compound of any one of claims 1, 32, and 35, having a structure selected from: 【Chemistry 18】

39. 37. The compound of any one of claims 1, 32, and 36, having a structure selected from: 【Chemistry 19】

40. Y ２ The compound of any one of claims 37 to 39, wherein is unsubstituted heteroaryl.

41. Y ２ but, 【Chemistry 20】 41. The compound of claim 40, selected from:

42. Y ２ but, 【Chemistry 21】 42. The compound of claim 41, wherein

43. Y ２ The compound of any one of claims 37 to 39, wherein is substituted heteroaryl.

44. Y ２ but, 【Chemical 22】 and R １０ , R １１ , and R １２ is -H, halogen, -CN, -OH 、 -NH ２ , -OCF ３ , -OCHF ２ , -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR １３ R １４、 -CO ２ R １５ and -C(O)NHSO ２ R １５ Independently selected from １０ , R １１ , and R １２ At least one of is not -H, R １３ , R １４ , and R １５ 44. The compound of claim 43, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

45. R １０ , R １１ , and R １２ is -H, -F, -Cl, -Br, -CN, -CH ３ , -CH ２ CH ３、 -CF ３ , -CHF ２ , -CF ２ CH ３、 -OCH ３ , -OCF ３ , -OCHF ２ , -OAc, -NH ２ , -NHCH ３ , -NHAc, -C(O)NH ２ , -C(O)NHCH ３ , -C(O)NHCH ２ CH ３ , -C(O)NHSO ２ CH ３ , -C(O)NHSO ２ CH ２ CH ３ , -CO ２ 45. The compound of claim 44, wherein each of the groups is independently selected from H, phenyl, cyclopropyl, cyclobutyl, imidazolyl, and tetrazolyl.

46. R １０ and R １２ are each -H, and R １１ But -CN, -CF ３ , -CH ３ , -OCH ３ , -NH ２ , -NHCH ３ , -NHAc, -CO ２ H, —C(O)NH ２ , -C(O)NHCH ３ , -C(O)NHCH ２ CH ３ , 【Chemistry 23】 46. ​​The compound of claim 45, selected from:

47. R １１ and R １２ are each -H, and R １０ But -CN, -CF ３ , -CH ３ , -OCH ３ , -NH ２ , -NHCH ３ , -NHAc, -CO ２ H, -C(O)NH ２ , -C(O)NHCH ３ , -C(O)NHCH ２ CH ３ , 【Chemistry 24】 46. ​​The compound of claim 45, selected from:

48. R １０ and R １１ are each -H, and R １２ But -CN, -CF ３ , -CH ３ , -OCH ３ , -NH ２ , -NHCH ３ , -NHAc, -CO ２ H, —C(O)NH ２ , -C(O)NHCH ３ , -C(O)NHCH ２ CH ３ , 【Chemistry 25】 46. ​​The compound of claim 45, selected from:

49. Y ２ but, 【Chemistry 26】 is selected from R １６ For each occurrence, halogen, -CN, -NH ２ , -OCF ３ , -OCHF ２ , -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR １３ R １４ , -CO ２ R １５ are independently selected from R １３ , R １４ , and R １５ 44. The compound of claim 43, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

50. R １６ -CN, -CH ３ , -CF ３ , -C(O)NH ２ , -CO ２ CH ２ CH ３ , and 【Chemistry 27】 50. The compound of claim 49, selected from:

51. Y ２ but, 【Chemistry 28】 is selected from R １７ , R １８ , R １９ , R ２０ , and R ２１ Each occurrence of is -H, halogen, -CN, -NH ２ , -OCF ３ , -OCHF ２ , -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR １３ R １４、 and -CO ２ R １５ Independently selected from １７ , R １８ , R １９ , R ２０ , and R ２１ At least one of is not -H, R １３ , R １４ , and R １５ 44. The compound of claim 43, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

52. R １７ , R １８ , R １９ , R ２０ , and R ２１ is -H, -CN, -CH ３ , and -OCH ３ 52. The compound of claim 51 , independently selected from:

53. Y ２ but, 【Chemical 29】 44. The compound of claim 43, selected from:

54. 35. The compound of any one of claims 1, 33, and 34, having a structure selected from: 【Chemistry 30】

55. 36. The compound of any one of claims 1, 33, and 35, having a structure selected from: 【Chemistry 31】

56. 37. The compound of any one of claims 1, 33, and 36, having a structure selected from: 【Chemistry 32】

57. Y ２ The compound of any one of claims 54 to 56, wherein is unsubstituted cycloalkyl or heterocyclyl.

58. Y ２ but, 【Chemical 33】 58. The compound of claim 57, selected from:

59. Y ２ but, 【Chemical 34】 58. The compound of claim 57, selected from:

60. Y ２ The compound of any one of claims 54 to 56, wherein is substituted cycloalkyl or heterocyclyl.

61. Y ２ but, 【Chemistry 35】 61. The compound of claim 60, selected from:

62. Y ２ 57. The compound of any one of claims 54 to 56, wherein is selected from alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, and hydroxyalkyl.

63. Y ２ But -CH ３ , -CH ２ CH (CH ３ ) ２ , -CH ２ CH ２ C≡CH, -CH ２ CH ２ OCH ３ , -C(H)(CH ３ ) CH ２ OCH ３ , -OCH ３ , -CH ２ OH, -CH ２ CH ２ OH, -C(CH ３ ) ２ OH and -CH ２ OCH ３ 63. The compound of claim 62, selected from:

64. Y ２ But -CH ２ OH and -CH ２ CH ２ 64. The compound of claim 63, wherein said compound is selected from: OH.

65. Y ２ The compound of any one of claims 54 to 56, wherein is heteroaryl.

66. Y ２ but, 【Chemical 36】 66. The compound of claim 65, selected from:

67. Y ２ but, 【Chemical 37】 and R １０ , R １１ , and R １２ is -H, halogen, -CN, -OH 、 -NH ２ , -OCF ３ , -OCHF ２ , -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR １３ R １４、 and -CO ２ R １５ are independently selected from R １３ , R １４ , and R １５ 66. The compound of claim 65, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

68. R １０ , R １１ , and R １２ The compound of claim 67, wherein at least one of is not -H.

69. Y ２ but, 【Chemical 38】 is selected from R １７ , R １８ , R １９ , R ２０ , and R ２１ Each occurrence of is -H, halogen, -CN, -NH ２ , -OCF ３ , -OCHF ２ , -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR １３ R １４、 and -CO ２ R １５ are independently selected from R １３ , R １４ , and R １５ 66. The compound of claim 65, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

70. R １７ , R １８ , R １９ , R ２０ , and R ２１ 70. The compound of claim 69, wherein at least one of is not --H.

71. Y ２ but, 【Chemical 39】 is selected from R ２２ , R ２３ , R ２４ , and R ２５ Each occurrence of is -H, halogen, -CN, -NH ２ , -OCF ３ , -OCHF ２ , -OAc, -NHAc, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, alkylamino, cycloalkyl, aryl, heteroaryl, -C(O)NR １３ R １４、 and -CO ２ R １５ are independently selected from R １３ , R １４ , and R １５ 66. The compound of claim 65, wherein each occurrence of is independently selected from -H, alkyl, aryl, and heteroaryl.

72. R ２２ , R ２３ , R ２４ , and R ２５ Each occurrence of is selected from -H, -CH ３ 72. The compound of claim 71, independently selected from:

73. Y ２ is -NH(Y ２ ') or Y ２ But -N(Y ２ '') ２ The compound according to any one of claims 54 to 56,

74. Y ２ 'が、-H、-OH、-CH ３ 、-CH ３ 、-CH ２ CH ２ OCH ３ 、 【Chemistry 40】 74. The compound of claim 73, selected from:

75. Each Y ２ '' is -CH ３ 74. The compound of claim 73, wherein:

76. Both Y's ２ 74. The compound of claim 73, wherein "" together with the nitrogen atom to which they are attached form morpholinyl.

77. Y ２ is -NH(Y ２ 57. The compound according to any one of claims 54 to 56, wherein

78. Y ２ 78. The compound of claim 77, wherein ' is selected from -H, alkyl, alkoxy, and hydroxyalkyl.

79. Y ２ ' is -H, -OCH ３ , -CH ３ , and -CH ２ CH ２ 79. The compound of claim 78, wherein said compound is selected from: OH.

80. Y ３ and Y ４ are both -H or both -F.

81. Y ３ But -F, -CF ３ , —OH and —OCH ３ is selected from Y ４ The compound of any one of claims 1 to 79, wherein is -H.

82. Y ４ But -F, -CF ３ , —OH and —OCH ３ is selected from Y ３ The compound of any one of claims 1 to 79, wherein is -H.

83. Y ５ and Y ６ are both -H or both -F.

84. Y ５ But -F, -CF ３ , —OH and —OCH ３ is selected from Y ６ The compound of any one of claims 1 to 79, wherein is -H.

85. Y ６ But -F, -CF ３ , —OH and —OCH ３ is selected from Y ５ The compound of any one of claims 1 to 79, wherein is -H.

86. A compound having the structure of any one of the following compounds or a pharma- ceutically acceptable salt thereof: 【Table 1】

87. A compound having the structure of any one of the following compounds or a pharma- ceutically acceptable salt thereof: 【Table 2】

88. A compound having the structure of any one of the following compounds or a pharma- ceutically acceptable salt thereof: 【Table 3】

89. A compound having the structure of any one of the following compounds or a pharma- ceutically acceptable salt thereof: 【Table 4】

90. A pharmaceutical composition comprising a compound according to any one of claims 1 to 89 and a pharma- ceutically acceptable excipient.

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 described in any one of claims 1 to 89.

92. 90. A method for treating or preventing phenylketonuria, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 89.

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 claims 1 to 89.

94. 94. The method of any one of claims 91 to 93, wherein the compound reduces phenylalanine levels in the body of the subject.

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 claims 1 to 89.

96. 96. The method of claim 95, wherein the compound decreases tyrosine levels in the subject's body.

97. 90. A method for treating or preventing nonketotic hyperglycinemia, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1-89.

98. 98. The method of claim 97, wherein the compound reduces glycine levels in the subject's body.

99. 90. A method for treating or preventing isovaleric acidemia, methylmalonic acidemia, propionic acidemia, maple syrup urine disease, DNAJC12 deficiency, a urea cycle disorder, or hyperammonemia, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 89.

100. 90. 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 claims 1 to 89.

101. The method of any one of claims 91 to 100, wherein the compound inhibits SLC6A19 in the subject.

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