Methods for treating erythropoietic protoporphyria, X-linked protoporphyria or congenital erythropoietic porphyria

Abstract

Embodiments of the present invention relate to methods of preventing or treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP) and / or congenital erythropoietic porphyria (CEP) and related syndromes using a glycine transporter inhibitor, such as a GlyT1 inhibitor, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, or a pharmaceutical composition thereof.

Description

[0001] This application is a divisional application of the Chinese patent application with application number 202180019091.1 (application date: January 8, 2021, invention name: Method for treating erythropoietic protoporphyria, X-linked protoporphyria or congenital erythropoietic porphyria with glycine transporter inhibitors).

[0002] Related applications

[0003] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 62 / 958,892, filed January 9, 2020, and U.S. Provisional Patent Application No. 62 / 085,942, filed September 30, 2020, which are hereby incorporated by reference in their entireties. Technical Field

[0004] Embodiments disclosed herein relate to methods and uses for preventing or treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP), or congenital erythropoietic porphyria (CEP) with a glycine transporter inhibitor, such as, but not limited to, a GlyT1 inhibitor, or a pharmaceutically acceptable salt, solvate, prodrug, or pharmaceutical composition thereof. Background Art

[0005] Erythropoietic protoporphyria (EPP) is a globally prevalent disease affecting approximately 5,000-10,000 individuals worldwide (Michaels et al. 2010). EPP is considered the most common form of porphyria in children. EPP is a form of porphyria that varies in severity and can be very painful. It results from a deficiency of the enzyme ferrochelatin, leading to abnormally high levels of protoporphyrin IX in red blood cells (erythrocytes), plasma, skin, and liver. Erythropoietic protoporphyria (EPP) is caused by an inherited or acquired deficiency in ferrochelatin activity. X-linked protoporphyria (XLPP) is caused by an inherited increase in the activity of delta-aminolevulinic acid synthase-2 (ALAS2). The enzymes that cause both EPP and XLPP are in the heme biosynthesis pathway. EPP and XLPP are clinically nearly identical. Congenital erythropoietic porphyria (CEP), also known as Gunther disease, is caused by mutations in the gene for uroporphyrinogen synthase that result in reduced activity of this enzyme and accumulation of the upstream metabolite coproporphyrin I. Current treatments for erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP), or congenital erythropoietic porphyria (CEP) are limited. Therefore, new methods and compositions for treating and / or preventing erythropoietic protoporphyria, X-linked protoporphyria, and congenital erythropoietic porphyria are needed. Methods and uses of glycine transporter inhibitors, such as, but not limited to, GlyT1 inhibitors, as described herein, meet these and other needs. Summary of the Invention

[0006] The present application provides a method for treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP), or congenital erythropoietic porphyria (CEP) in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter 1 (GlyT1) inhibitors or pharmaceutically acceptable salts thereof, or prodrugs of the one or more GlyT1 inhibitors or their salts.

[0007] The present application also provides a method for preventing, treating, or reducing the progression rate and / or severity of one or more complications of EPP, XLPP, or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more GlyT1 inhibitors or pharmaceutically acceptable salts thereof, or prodrugs of the one or more GlyT1 inhibitors or pharmaceutically acceptable salts thereof. In certain embodiments, the one or more complications of EPP, XLPP, or CEP are selected from the group consisting of acute photosensitivity, cutaneous photosensitivity, edema, erythema, anemia, hypochromic anemia, hemolytic anemia, hemolysis, mild hemolysis, severe hemolysis, chronic hemolysis, hypersplenism, palmar keratosis, bullae, lesions, scars, deformities, nail loss, toe loss, cholestasis, cell lysis, gallstones, cholestatic liver failure, cholelithiasis, mild liver disease, exacerbated liver disease, end-stage liver disease, erythrodontia, hypercellular bone marrow, myelodysplasia, thrombocytopenia, fetal hydrops, and / or intrauterine death. In certain such embodiments, the acute photosensitivity is due to sun exposure.

[0008] The present application further provides a method for preventing or treating EPP, XLPP or CEP in a subject, wherein the method comprises administering to the subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of one or more GlyT1 inhibitors or a pharmaceutically acceptable salt thereof.

[0009] The present application further provides a method for preparing a medicament for treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject at least one GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the one or more GlyT1 inhibitors or a pharmaceutically acceptable salt thereof.

[0010] The present application further provides a method for preparing a medicament for inhibiting protoporphyrin IX (PPIX) synthesis in vivo, comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of one or more GlyT1 inhibitors or a pharmaceutically acceptable salt thereof.

[0011] In certain embodiments, the subject has EPP. In other embodiments, the subject has XLPP. In still other embodiments, the subject has CEP.

[0012] In certain embodiments, the methods increase pain-free light exposure in a subject. In other embodiments, the methods decrease light sensitivity in a subject.

[0013] The present application further provides a method for inhibiting PPIX synthesis in vivo, comprising administering a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof to a subject.

[0014] The present application further provides a method for inhibiting zinc protoporphyrin IX (ZPPIX) synthesis in vivo, comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof.

[0015] The present application further provides a method for inhibiting the synthesis of uroporphyrin I and / or coproporphyrin I in vivo, comprising administering a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof to a subject.

[0016] The present application further provides a method for inhibiting 5-aminolevulinic acid (5-ALA) synthesis in vivo, comprising administering a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof to a subject.

[0017] In certain embodiments, the accumulation of one or more heme intermediates is inhibited, and wherein the one or more heme intermediates are selected from PPIX, ZPPIX, uroporphyrin I, coproporphyrin I and / or 5-ALA. In certain such embodiments, the accumulation of the one or more heme intermediates is inhibited in a dose-dependent manner.

[0018] In certain embodiments, the GlyT1 inhibitor exhibits an EC50 of less than 500 nM. In certain embodiments, the GlyT1 inhibitor exhibits an EC50 of less than 100 nM.

[0019] In certain embodiments, at least 50% cell viability is maintained. In certain embodiments, at least 90% cell viability is maintained.

[0020] In certain embodiments, prior to administration of the GlyT1 inhibitor, the subject's PPIX level is at least 10%, 20%, 30%, 40%, or 50% greater than the PPIX level of a healthy subject.

[0021] In certain embodiments, prior to administration of the GlyT1 inhibitor, the subject's ZPPIX levels are at least 10%, 20%, 30%, 40%, or 50% greater than the ZPPIX levels of healthy subjects.

[0022] In certain embodiments, the subject has an increased ratio of ZPPIX to free protoporphyrin IX (ZPPIX / PPIX ratio) compared to a subject with EPP.

[0023] In certain embodiments, prior to administration of the GlyT1 inhibitor, the subject's uroporphyrin I and / or coproporphyrin I levels are at least 10%, 20%, 30%, 40%, or 50% higher than uroporphyrin I and / or coproporphyrin I levels in healthy subjects.

[0024] In certain embodiments, prior to administration of the GlyT1 inhibitor, the subject's 5-ALA levels are at least 10%, 20%, 30%, 40%, or 50% higher than the 5-ALA levels of healthy subjects.

[0025] In certain embodiments, the PPIX level of the subject is reduced, and the hemoglobin level of the patient is significantly maintained. In certain embodiments, the PPIX level of the patient is reduced by at least 50% (for example, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%) and the hemoglobin level of the patient is reduced by no more than 10% (for example, 10%, 15%, 20%, 25% and 30%). In certain embodiments, the PPIX level of the patient is reduced by at least 85% and the hemoglobin level of the patient is reduced by no more than 15%. In certain embodiments, the hemoglobin level is reduced by no more than 10% (for example, 10%, 15%, 20%, 25% and 30%). In certain embodiments, the dosage of the pharmaceutical composition does not cause a significant reduction in hemoglobin level.

[0026] In certain embodiments, the free protoporphyrin IX level in the erythrocyte of experimenter increases.In certain embodiments, described method reduces the free protoporphyrin IX level of experimenter.In some such embodiments, described method makes the free protoporphyrin IX level of experimenter reduce by at least 10% (for example, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%).In certain embodiments, the protoporphyrin IX level increases in the feces of experimenter.In certain embodiments, described method reduces the protoporphyrin IX level in the feces of experimenter. In certain such embodiments, the method reduces the level of protoporphyrin IX in the feces of the subject by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0027] In certain embodiments, when irradiated with blue light (e.g., 400-420nm light), the plasma porphyrin of the experimenter fluoresces at a peak value of 634nm. In certain embodiments, when irradiated with blue light (e.g., 400-420nm light), the plasma porphyrin of the experimenter fluoresces at a peak value of 626nm and 634nm. In certain embodiments, when irradiated with blue light (e.g., 400-420nm light), the skin porphyrin of the experimenter fluoresces at a peak value of 632nm. In certain embodiments, when irradiated with blue light (e.g., 400-420nm light), the skin porphyrin of the experimenter fluoresces at a peak value of 626nm and 634nm.

[0028] In certain embodiments, protoporphyrin IX level increases in the skin of the experimenter. In certain embodiments, described method reduces the protoporphyrin IX level in the skin of the experimenter. In certain embodiments, described method reduces the protoporphyrin IX level in the skin of the experimenter. In some such embodiments, described method reduces the protoporphyrin IX level in the skin of the experimenter by at least 10% (for example, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%). In certain embodiments, protoporphyrin IX level is greater than 0.2 fluorescence unit (FDU) in the skin of the experimenter. In certain embodiments, protoporphyrin IX level is greater than 1.0FDU in the skin of the experimenter. In certain embodiments, the protoporphyrin IX level in the skin of the experimenter is between 1.0FDU and 2.5FDU. In certain embodiments, protoporphyrin IX level is greater than 2.5FDU in the skin of the experimenter. In certain embodiments, the method reduces the level of protoporphyrin IX in the skin of a subject to less than 0.5 FDU. In certain embodiments, the method reduces the level of protoporphyrin IX in the skin of a subject to less than 1.0 FDU. In certain embodiments, the method reduces the level of protoporphyrin IX in the skin of a subject to less than 1.5 FDU. In certain embodiments, the method reduces the level of protoporphyrin IX in the skin of a subject to less than 2.0 FDU. In certain embodiments, the method reduces the level of protoporphyrin IX in the skin of a subject to less than 2.5 FDU.

[0029] In certain embodiments, the protoporphyrin IX level in the erythrocyte of experimenter increases.In certain embodiments, described method reduces the protoporphyrin IX level in the erythrocyte of experimenter.In some such embodiments, described method makes the protoporphyrin IX level in the erythrocyte of experimenter reduce by at least 10% (for example, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%). In certain embodiments, the protoporphyrin IX level in the erythrocyte of experimenter is greater than 31 μ mol L -1 In certain embodiments, the protoporphyrin IX level in the subject's erythrocytes is between 31 μmol L -1 With 53 μmol L -1 In certain embodiments, the protoporphyrin IX level in the subject's erythrocytes is greater than 53 μmol L -1 In certain embodiments, the method reduces the level of protoporphyrin IX in the subject's red blood cells to less than 53 μmol L -1 In certain embodiments, the method reduces the level of protoporphyrin IX in the subject's red blood cells to less than 31 μmol L -1 In certain embodiments, the method reduces the level of protoporphyrin IX in the subject's red blood cells to less than 15 μmol L -1 level.

[0030] In certain embodiments, the subject's ferrochelatase activity level is reduced to between 10% and 35% of the ferrochelatase activity level observed in normal subjects. In certain embodiments, the subject's ferrochelatase activity level is reduced to less than 50% of the ferrochelatase activity level observed in normal subjects.

[0031] In certain embodiments, the subject has a gain-of-function mutation in ALAS2. In certain embodiments, the subject has increased ALAS2 enzyme activity.

[0032] In certain embodiments, the zinc protoporphyrin IX level in the erythrocyte of the experimenter increases.In certain embodiments, the method reduces the zinc protoporphyrin IX level in the erythrocyte of the experimenter.In some such embodiments, the method reduces the zinc protoporphyrin IX level in the erythrocyte of the experimenter by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%).

[0033] In certain embodiments, the uroporphyrinogen III synthase activity of the experimenter is reduced. In certain embodiments, the uroporphyrin I and / or coproporphyrin I levels of the experimenter are increased. In certain embodiments, the uroporphyrin I and / or coproporphyrin I levels of increase are measured in the urine or red blood cells of the experimenter. In certain embodiments, the coproporphyrin I levels of increase are measured in the feces of the experimenter. In certain embodiments, the method reduces the uroporphyrin I and / or coproporphyrin I levels of the experimenter. In certain embodiments, the method reduces the uroporphyrin I levels of the experimenter. In certain embodiments, the method reduces the uroporphyrin I levels of the experimenter. In certain embodiments, the method reduces the uroporphyrin I levels of the experimenter by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%). In certain embodiments, the methods reduce the level of coproporphyrin I in the subject. In certain embodiments, the methods reduce the level of coproporphyrin I in the subject by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0034] In certain embodiments, the subject has a mutation in UROS.

[0035] In certain embodiments, the subject has a genetic defect in the GATA-1 erythroid-specific transcription factor.

[0036] In certain embodiments, the subject has red fluorescent urine.In certain embodiments, the subject has a peak between 615 nm and 620 nm using plasma porphyrin fluorescence analysis.

[0037] In certain embodiments, the subject has a liver disease associated with EPP, XLPP, or CEP. In certain embodiments, the liver disease associated with EPP, XLPP, or CEP is cholelithiasis. In certain embodiments, the liver disease associated with EPP, XLPP, or CEP is mild liver disease. In certain embodiments, the liver disease associated with EPP, XLPP, or CEP is advanced liver disease. In certain embodiments, the liver disease associated with EPP, XLPP, or CEP is end-stage liver disease.

[0038] In certain embodiments, the method further comprises administering to the subject an additional active agent and / or supportive therapy. In certain such embodiments, the additional active agent and / or supportive therapy is selected from the group consisting of: sun avoidance, topical sunscreen, skin protection, UVB phototherapy, afamelanotide Bortezomib, proteasome inhibitors, chemical chaperones, cholestyramine, activated charcoal, iron supplementation, liver transplantation, bone marrow transplantation, splenectomy, and blood transfusion.

[0039] In certain embodiments, the GlyT1 inhibitor is a compound having formula I wherein Ar is an unsubstituted or substituted aryl or a 6-membered heteroaryl containing 1, 2 or 3 nitrogen atoms, wherein the substituted aryl and the substituted heteroaryl are substituted by one or more substituents selected from the group consisting of hydroxy, halogen, NO2, CN, (C1-C6)-alkyl, (C1-C6)-alkyl substituted by halogen, (C1-C6)-alkyl substituted by hydroxy, (CH2)n-(C1-C6)-alkoxy, (C1-C6)-alkoxy substituted by halogen, NR 7 R 8 、C(O)R 9 、SO2R 10 and -C(CH3)=NOR 7 , or substituted by a 5-membered aromatic heterocyclic ring containing 1 to 4 heteroatoms selected from N and O, said aromatic heterocyclic ring being optionally substituted by a (C1-C6)-alkyl group; R 1 is hydrogen or (C1-C6)-alkyl; R 2 is hydrogen, (C1-C6)-alkyl, (C2-C6)-alkenyl, (C1-C6)-alkyl substituted by halogen, (C1-C6)-alkyl substituted by hydroxy, (CH2)n-(C3-C7)-cycloalkyl, CH(CH3)-(C3-C7)-cycloalkyl, (CH2) n+1 -C(O)-R 9 、(CH2) n+1 -CN, bicyclo[2.2.1]heptyl, (CH2) n+1 -O-(C1-C6)-alkyl, (CH2) n -heterocycloalkyl, (CH2) n -aryl or (CH2) containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur or nitrogen n -5- or 6-membered heteroaryl, wherein aryl, heterocycloalkyl and heteroaryl are unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, (C1-C6)-alkyl and (C1-C6)-alkoxy; R 3 、R 4 and R 6 are each independently hydrogen, hydroxy, halogen, (C1-C6)-alkyl, (C1-C6)-alkoxy or O-(C3-C6)-cycloalkyl; R 5 NO2, CN, C(O)R 9 or SO2R 10 ; R7 and R 8 are each independently hydrogen or (C1-C6)-alkyl; R 9 is hydrogen, (C1-C6)-alkyl, (C1-C6)-alkoxy or NR 7 R 8 ; R 10 is (C1-C6)-alkyl, (CH2) optionally substituted by halogen n -(C3-C6)-cycloalkyl, (CH2) n -(C3-C6)-alkoxy, (CH2) n -heterocycloalkyl or NR 7 R 8 ; n is 0, 1 or 2; or a pharmaceutically acceptable salt thereof or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0040] In certain embodiments, the GlyT1 inhibitor is a compound having the formula The compound bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0041] In certain embodiments, the GlyT1 inhibitor is Compounds of formula II, wherein R1 represents a heteroaryl group selected from the group consisting of imidazolyl, thiazolyl, pyridinyl, oxazolyl, pyrazolyl, triazolyl, oxadiazolyl, quinolinyl, isoxazolyl, pyrroloimidazolyl, and thiadiazole, wherein the heteroaryl group is optionally substituted with one or more substituents selected from the group consisting of -OH, -NR7R8, halogen, (C1-C8)alkyl, (C3-C 10 )cycloalkyl, (C1-C8)alkoxy, (C1-C 12 ) alkoxyalkyl, (C1-C8) hydroxyalkyl, (C6-C 14 ) aryl and benzyl; R2, R3 and A independently represent H or (C1-C8) alkoxy, wherein the alkyl is optionally substituted by one or more -OH, (C1-C8) alkoxy, -NR7R8 or halogen; Q represents -(CH2) n -, wherein n=1, 2, 3 or 4 or -(CH2) m -O-, wherein m=2, 3 or 4; Z represents (C6-C 14 ) aryl, (C1-C8) alkyl or (C3-C8) cycloalkyl; R4 and R5 each independently represent H, halogen, (C1-C8) alkyl, (C6-C 14 )aryl, (C6-C 14)aryloxy, (C1-C8)alkoxy, (3-10 membered)heterocycloalkyl or (C3-C8)cycloalkoxy; wherein R4 and R5 are optionally substituted by one or more -OH, (C1-C8)alkoxy, -NR7R8 or halogen; Y represents -R6, -(CH2)o-R6, -C(R6)3 or -CH(R6)2, wherein O=1, 2 or 3; R6 represents H, (C6-C 14 ) aryl, (C 1-10 )alkyl, (C3-C 10 )cycloalkyl, (C5-C 18 )bicycloalkyl, (C5-C 18 )tricycloalkyl, (3-10 membered)heterocycloalkyl, (5-10 membered)heteroaryl, -C(=O)NR7R8 or -C(=O)OR7, wherein the R6 group may be optionally substituted with one or more X groups; wherein X = -OH, (C1-C8)alkoxy, -NR 11 R 12 、-SO2R 10 、-C(=O)R 10 , halogen, cyano, (C1-C8) alkyl, (C1-C 10 )alkoxyalkyl, (5-10 membered)heteroaryl, (C6-C 14 )aryl, (C6-C 14 )aryloxy, benzyl or (C1-C8)hydroxyalkyl; wherein R7 and R8 independently represent H, (C1-C8)alkyl, (C3-C8)cycloalkyl, (5-10 membered)heterocycloalkyl, (C1-C8)hydroxyalkyl, (5-10 membered)heteroaryl or (C1-C 10 )alkoxyalkyl; wherein R7 and R8 may be optionally substituted with one or more X groups; or R7 and R8 together with the nitrogen to which they may be attached may form a (3-10 membered) heterocycloalkyl optionally substituted with one or more X groups; wherein R 10 represents (C1-C8)alkyl, (C3-C8)cycloalkyl, (3-10 membered)heterocycloalkyl, (C1-C8)hydroxyalkyl, (5-10 membered)heteroaryl or (C1-C 10 ) alkoxyalkyl; wherein R 11 and R 12 independently represents H, (C1-C8)alkyl, (C3-C8)cycloalkyl, (5-10 membered)heterocycloalkyl, (C1-C8)hydroxyalkyl, (5-10 membered)heteroaryl or (C1-C8) 10 ) alkoxyalkyl; or a pharmaceutically acceptable salt thereof or a prodrug of said compound or a pharmaceutically acceptable salt thereof. In certain such embodiments, the GlyT1 inhibitor is a compound having the formula In other such embodiments, the GlyT1 inhibitor is a compound of formula The compound PF-3463275 or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0042] In certain embodiments, the GlyT1 inhibitor is a compound of formula III A compound of the formula wherein Z 1 Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, halo C 1-4 Alkyl, phenyl, halogenated C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkylsulfinyloxy, C 1-4 Alkylsulfonyl, bromine and chlorine; Z 2 Selected from hydrogen, halogen, cyano, C 1-4 Alkyl, phenyl, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkoxy C 1-4 Alkyl and C 3-6 Cycloalkyl; Z 3 Selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, halo C 1-4 Alkyl, halogenated C 1-4 Alkoxy and C 3-6 Cycloalkyl; Z 4 Selected from hydrogen, halogen, C1-3 alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, phenyl, halo C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkoxy C 1-4 Alkyl and C 3-6 Cycloalkyl; Z 5 Selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, phenyl, halo C 1-4 Alkyl, halogenated C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkoxy C 1-4 Alkyl and C 3-6 Cycloalkyl; wherein if Z 1to Z 5 If more than one of them is a methoxy group, then only Z 1 and Z 5 is methoxy, R 3 and R 4 independently selected from hydrogen and C optionally substituted by one or more groups Y 1-4 Alkyl; or R 3 and R4 together with the nitrogen atom to which they are attached form a saturated or partially unsaturated A, 5, 6 or 7 membered carbocyclic ring optionally substituted by a group Y'; Y is selected from C 1-4 Alkoxy, hydroxy, halogenated C 1-4 Alkoxy and C 3-5 Cycloalkyl; Y' is selected from C 1-4 Alkyl, C 1-4 Alkoxy, halogen, hydroxyl, halogenated C 1-4 Alkoxy, C 3-5 Cycloalkyl and C 5-10 Aryl, or Y' forms a -CH2- or -CH2-CH2- bridge between two atoms on A, a 5-, 6-, or 7-membered carbon ring; R 5 and R 6 is independently C optionally substituted with one or more groups X 1-4 Alkyl; or R 5 and R 6 Together with the carbon atoms to which they are attached, they form a saturated 5- or 6-membered carbocyclic ring optionally substituted by one or more groups X', in which R 5 In the case where R and R6 together with the carbon atom to which they are attached form a 5-membered saturated carbocyclic ring, the ring may optionally further comprise an additional heteroatom group selected from O, N and S(O)m, wherein m=0, 1 or 2; X is selected from halogen, hydroxy, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy and C 5-10 Aryl; and X' is selected from halogen, hydroxy, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy and C 5-10 Aryl; wherein R 3 、R 4 、R 5 and R 6 are not all unsubstituted methyl groups at the same time; provided that when Z 1 is a propoxy group, Z 3 It is chlorine, Z 2 =Z 4 =Z 5 =H, and R 5 and R 6When both are methyl groups, then R 3 and R 4 Together with the nitrogen atom to which they are attached, they do not form a 2-methylpyrrolidino group; when Z 1 is methyl, Z 3 is methoxy, Z 2 =Z4=Z5=H, and R 5 and R 6 When both are methyl groups, then R 3 and R 4 Together with the nitrogen atom to which they are attached, they do not form a pyrrolidine group, or a pharmaceutically acceptable salt thereof or a prodrug of said compound or a pharmaceutically acceptable salt thereof. In certain such embodiments, the GlyT1 inhibitor is of the formula A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0043] In certain embodiments, the GlyT1 inhibitor is a compound of formula IV A compound of the formula wherein Z is (CH2) n , O, S, SO, SO2 or N-R5; n is 0, 1 or 2; X represents 1-3 independently selected from hydrogen, halogen, (C 1-6 ) alkoxy, (C 3-6 )cycloalkoxy, (C 6-12 ) aryloxy, (C 6-12 )aryl, thienyl, SR6, SOR6, SO2R6, NR6R6, NHR6, NH2, NHCOR6, NSO2R6, CN, COOR6 and (C 1-4 )alkyl, which is optionally substituted by halogen, (C 6-12 ) aryl, (C 1-6 ) alkoxy or (C 6-12 ) aryloxy substituted; or two substituents at adjacent positions together represent a fused (C 5-6 ) aryl, fused (C 5-6 )cycloalkyl ring or O-(CH2) m -O; m is 1 or 2; Y represents 1-3 independently selected from hydrogen, halogen, (C 1-4 ) alkoxy, SR6, NR6R6 and (C 1-4 ) alkyl substituents, optionally substituted by halogen; R1 is COOR7 or CONR8R9; R2 and R6 are (C 1-4 ) alkyl; R3, R4 and R5 are independently hydrogen or (C 1-4 ) alkyl; R7, R8 and R9 are independently hydrogen, (C 1-4 )alkyl, (C 6-12) aryl or aralkyl, or a pharmaceutically acceptable salt thereof or a prodrug of said compound or a pharmaceutically acceptable salt thereof. In certain such embodiments, the GlyT1 inhibitor is a compound having the formula

[0044] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0045] In certain embodiments, the GlyT1 inhibitor is

[0046] With formula V A compound of the formula wherein n is an integer from 1 to 3; R 1 and R 2 are independently selected from hydrogen, alkyl, haloalkyl, alkoxy, haloalkoxy, aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein the above ring is optionally replaced by R a 、R b or R c Substituted, the R a 、R b or R c R is independently selected from alkyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxy, cyano, monosubstituted amino or disubstituted amino; or 1 and R 2 When attached to the same carbon atom, they can combine to form a cycloalkyl or monocyclic saturated heterocyclic group to give a spirocycle, wherein the cycloalkyl or monocyclic saturated heterocyclic group can be optionally replaced by R d 、R c or R f Substituted, the R d 、R c or R f R is independently selected from alkyl, alkoxy, fluorine, fluoroalkyl, fluoroalkoxy, hydroxy, monosubstituted amino or disubstituted amino; or 1 and R 2 When attached to the carbon atoms at positions 2 and 5 or 3 and 6 of the piperazine ring, they can combine to form a -C1-C3-alkylene chain, wherein one carbon atom in the alkylene chain is optionally replaced by -NR-, -O-, -S(O)n- (wherein R is hydrogen or alkyl and n is 0-2), and further wherein one or two hydrogen atoms in the alkylene chain can be optionally replaced by one or two alkyl groups; R 3 、R 4 and R 5 are independently hydrogen, alkyl, fluorine or fluoroalkyl; and Ar 1 and Ar 2 are independently aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein each of the above rings is optionally replaced by R g 、R h or Ri substituted, where Rg is an alkyl group, -C=CR 6 (where R 6 is aryl or heteroaryl), halogen, haloalkyl, haloalkoxy, alkylthio, cyano, alkoxy, amino, monosubstituted amino, disubstituted amino, sulfonyl, acyl, carboxyl, alkoxycarbonyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, aminosulfonyl, aminocarbonyl or acylamino, and R h and R i are independently selected from alkyl, halogen, haloalkyl, haloalkoxy, alkylthio, cyano, alkoxy, amino, monosubstituted amino, disubstituted amino, sulfonyl, acyl, carboxyl, alkoxycarbonyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, aminosulfonyl, aminocarbonyl, acylamino, aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein R g 、R h and R i The aromatic or alicyclic ring in is optionally replaced by R j 、R k or R l substituted, said Rj, Rk or Rl are independently selected from alkyl, halogen, haloalkyl, haloalkoxy, alkylthio, cyano, alkoxy, amino, monosubstituted amino, disubstituted amino, sulfonyl, acyl, carboxyl, alkoxycarbonyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, aminosulfonyl, aminocarbonyl or acylamino; or a pharmaceutically acceptable salt thereof, provided that: the compound of formula V is not 2-(4-diphenylmethylpiperazin-1-yl)acetic acid, 2-(4-( (4-chlorophenyl)(phenyl)methyl)piperazin-1-yl)acetic acid, 2-((2R,5S)-4-((R)-(4-(1H-tetrazol-5-yl)phenyl)(3-hydroxyphenyl)methyl)-2,5-dimethylpiperazin-1-yl)acetic acid, or 2-((2R,5S)-4-((R)-(4-cyanophenyl)(3-hydroxyphenyl)methyl)-2,5-dimethylpiperazin-1-yl)acetic acid, or a pharmaceutically acceptable salt thereof or a prodrug of said compound or a pharmaceutically acceptable salt thereof. In certain such embodiments, the GlyT1 inhibitor is a compound having the formula A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0047] In certain embodiments, the GlyT1 inhibitor is a compound having Formula VI A compound of the formula wherein A represents a group of the general formula N-R1, a group of the general formula N+(O-)R1 or a group of the general formula N+(R')R1, and wherein R1 represents a hydrogen atom, or a straight or branched (C1-C7)alkyl group optionally substituted with one or more fluorine atoms, or a (C4-C7)cycloalkyl group, or a (C3-C7)cycloalkyl (C1-C3)alkyl group, or a phenyl (C1-C3)alkyl group optionally substituted with one or two hydroxyl or methoxy groups, or a (C2-C4)alkenyl group, or a (C2-C4)alkynyl group; R' represents a straight or branched (C1-C7)alkyl group; X represents a hydrogen atom or one or more selected from halogen atom and trifluoromethyl, linear or branched (C1-C4) alkyl and (C1-C4) alkoxy substituents; R2 represents a hydrogen atom, or one or more substituents selected from halogen atoms and trifluoromethyl, (C1-C4) alkyl or (C1-C4) alkoxy, or an amino group of the general formula NR3R4, wherein R3 and R4 each independently represent a hydrogen atom or (C1-C4) alkyl, or form a pyrrolidine, piperidine or morpholine ring together with the nitrogen atom carrying them, or a phenyl group optionally substituted by an atom or group as defined above for symbol X, or a pharmaceutically acceptable salt thereof or a prodrug of said compound or a pharmaceutically acceptable salt thereof. In certain such embodiments, the GlyT1 inhibitor is a compound having the formula

[0048] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0049] In certain embodiments, the GlyT1 inhibitor is a compound having Formula VII

[0050] A compound of the formula wherein R 1 Yes - (CH2) n -R 1a , wherein n is independently 0-6, and R 1a Selected from: (1) C 1-6 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxyl, (2) R 2a 、R 2b and R 2c Substituted phenyl, (3)C 3-6 Cycloallyl, which is unsubstituted or replaced by C 1-6 Alkyl, 1-6 halogens, hydroxyl or -NR 10 R 11 Substitution, (4)-OC 1-6 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 Substitution, (5)-CO2R 9 , wherein R9 is independently selected from: (a) hydrogen, (b) -C1-6 alkyl, which is unsubstituted or substituted with 1 to 6 fluorine groups, (c) benzyl, and (d) phenyl,

[0051] (6)-NR 10 R 11 , where R 10 and R 11 are independently selected from: (a) hydrogen, (b) -C 1-6 Alkyl, which is unsubstituted or substituted by hydroxy, 1-6 fluorine or -NR 12 R 13 Substituted, where R 12 and R 13 are independently selected from hydrogen and -C 1-6 Alkyl, (c)-C 3-6 Cycloalkyl, which is unsubstituted or substituted by hydroxy, 1-6 fluorine or -NR 12 R 13 substituted, (d) benzyl, (e) phenyl, and (7) -CONR 10 R 11 ; R2 is selected from: (1) phenyl, which is R 2a 、R 2b and R 2c Replacement, (2) C 1-8 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy, -NR 10 R 11 , phenyl or heterocyclic ring, wherein the phenyl or heterocyclic ring is replaced by R 2a 、R 2b and R 2c Replacement, (3)C 3-6 Cycloalkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 Substitution, and (4)-C 1-6 Alkyl-(C 3-6 cycloalkyl), which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 Replacement; R 2a 、R 2b and R 2c Independently selected from: (1) hydrogen, (2) halogen, (3) -C 1-6 Alkyl, which is unsubstituted or substituted with: (a) 1-6 halogens, (b) phenyl, (c) C 3-6 Cycloalkyl or (d)-NR 10 R 11 , (4)-OC 1-6 Alkyl, which is unsubstituted or substituted with 1 to 6 halogens, (5) hydroxy, (6) -SCF3, (7) -SCHF2, (8) -SCH3, (9) -CO2R9 , (10)-CN, (11)-SO2R 9 ,(12)-SO2-NR 10 R 11 , (13)-NR 10 R 11 ,(14)-CONR 10 R 11 and (15)-NO2; R 3 Selected from: (1) C 1-6 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 Replacement, (2) C 3-6 Cycloalkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 Replacement, R 4 and R 5 are independently selected from: (1) hydrogen, and (2) C 1-6 Alkyl, which is unsubstituted or substituted by halogen or hydroxy, or R 4 and R 5 Together they form C 3-6 Cycloalkyl ring; A is selected from: (1) -O-, and (2) -NR 10 -; m is 0 or 1, wherein when m is 0, R 2 directly attached to the carbonyl group; and pharmaceutically acceptable salts thereof and individual enantiomers and diastereomers thereof, or pharmaceutically acceptable salts thereof or prodrugs of said compounds or pharmaceutically acceptable salts thereof. In certain such embodiments, the GlyT1 inhibitor is a compound having the formula

[0052] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0053] In certain embodiments, the GlyT1 inhibitor is a compound of formula VIII

[0054] A compound of the formula wherein R 1 are independently halogen, C1-C3 alkyl, C3-C6 cycloalkyl, OR 9 or SR 10 Phenyl substituted 1 to 5 times, wherein C1-C3 alkyl and C3-C6 cycloalkyl are optionally replaced by R 7 Replaced 1 to 10 times; R 2 It is H; R 3 and R4 are each independently H or CH3; R 5 Selected from: (1) hydrogen, (2) optionally replaced by R 7C1-C6 alkyl substituted 1 to 11 times, (3) geminal dialkyl, and (4) geminal dihalogen; or two R on the same carbon 5 The substituents, together with the carbon atom to which they are attached, may form a 7 3-, 4-, or 5-membered cycloalkyl substituted 1 to 10 times; or two R on adjacent carbons of the ring to which they are attached 5 The substituents may together form an optionally R 7 3-, 4-, 5- or 6-membered cycloalkyl substituted 1 to 10 times; R 6 yes wherein E, F and G are each independently nitrogen or carbon, and R 6a is C1-C2 alkyl, which is optionally substituted 1 to 5 times by halogen or deuterium; R 7 Selected from: (1) hydrogen, (2) halogen, (3) deuterium, (4) geminal dialkyl, (5) geminal dihalogen, (6) -OR 9 、-NR 11 R 12 、-NR 11 C(O) p R 10 、-S(O) p R 10 、-CN、-NO2、-C(O) p R 10 、-C(O)NR 11 R 12 or -NR 11 C(S)R 10 , and (7) oxo or thio; R 8 is selected from the group consisting of: (1) hydrogen, (2) halogen, (3) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl or C4-C7 cycloalkylalkyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each independently and optionally replaced by R 7 Substituted 1 to 11 times, or (4)-OR 9 、-NR 11 R 12 、-NR 11 C(O) p R 10 、-S(O) p R 10 、-CN、-NO2、-C(O) p R 10 、-C(O)NR 11 R 12 or -NR 11 C(S)R 10 ; R 9Selected from hydrogen, C1-C4 alkyl, C3-C7 cycloalkyl, C4-C7 cycloalkylalkyl, -C(O)NR 11 R 12 and -C(O) p R 10 , wherein C1-C4 alkyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each optionally replaced by R 7 Substituted 1 to 11 times; R 10 is selected from hydrogen, C1-C4 alkyl, C3-C7 cycloalkyl, C4-C7 cycloalkylalkyl, aryl and heteroaryl, wherein C1-C4 alkyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each optionally substituted 1 to 11 times with substituents as defined in R7, and aryl or heteroaryl is optionally substituted with R 8 Replaced 1 to 10 times; R 11 and R 12 are each independently selected from hydrogen, C1-C4 alkyl, C3-C7 cycloalkyl, C4-C7 cycloalkylalkyl, aryl and heteroaryl, wherein C1-C4 alkyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each optionally replaced by R 7 substituted 1 to 11 times by the substituents defined in 8 Replace 1 to 10 times, or R 11 and R 12 Together with the nitrogen to which they are attached, they form a 7 A is a saturated or partially saturated monocyclic or fused bicyclic heterocyclic ring substituted 1 to 11 times; X is N; Y is N; p is 1 or 2; and m is 0; provided that: R 6 It cannot be (a) 1H-1,2,3-triazol-4-yl, or (b) 5-methylisoxazol-4-yl; or an oxide thereof, a pharmaceutically acceptable salt of said compound or its oxide, or an individual enantiomer or diastereomer thereof.

[0055] In certain embodiments, the GlyT1 inhibitor is a compound having the formula

[0056]

[0057]

[0058] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0059] In certain embodiments, the GlyT1 inhibitor is a compound of Formula IX, where R 1represents a phenyl group or a 5- or 6-membered monocyclic heteroaryl group having 1, 2 or 3 heteroatoms independently selected from O, N or S, wherein the phenyl group or heteroaryl group is optionally replaced by one or more R 3 Replacement; R 2 represents an aryl group, a 5- or 6-membered monocyclic heteroaryl group or an 8- to 10-membered bicyclic heteroaryl group, said monocyclic or bicyclic heteroaryl group having 1, 2 or 3 heteroatoms independently selected from O, N or S, wherein said aryl or heteroaryl group is optionally replaced by one or more R 4 Replacement; R 3 Halogen, C 1-4 Alkyl or C 3-6 Cycloalkyl, wherein C 1-4 Alkyl or C 3-6 Cycloalkyl is optionally substituted with one or more halogens; and R 4 Is halogen, -CN, C 1-4 Alkyl, C 3-6 Cycloalkyl, -C 1-3 Alkyl-C 3-6 Cycloalkyl or -OC 1-6 Alkyl, where C 1-4 Alkyl, C 3-6 Cycloalkyl, -C 1-3 Alkyl-C 3-6 Cycloalkyl or -OC 1-6 The alkyl group is optionally substituted with one or more halogens; or a pharmaceutically acceptable salt thereof, or a tautomer or stereoisomer of the compound or a pharmaceutically acceptable salt thereof, or a mixture of any of the foregoing.

[0060] In certain embodiments, the GlyT1 inhibitor is a compound of formula X, where R 1 is selected from a) 5 or 6 membered monocyclic heteroaryl having 1, 2, 3 or 4 heteroatoms independently selected from O, N and S(O)r, b) 5 or 6 membered monocyclic partially saturated heterocycloalkyl having 1, 2 or 3 heteroatoms independently selected from O, N and S(O)r, and c) 9 or 10 membered bicyclic heteroaryl having 1, 2 or 3 heteroatoms independently selected from O, N and S(O)r. r wherein r is 0, 1 or 2; wherein the groups a), b) and c) are each optionally substituted by one or more substituents independently selected from the group consisting of: C 1-4 -alkyl-, C 1-4 -alkyl-O-, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, C 3-6 -cycloalkyl- and C 3-6 -cycloalkyl-O-, and in the case where a substituent is attached to a nitrogen ring atom, said substituent is selected from C 1-4 -alkyl-, C 1-4 -alkyl-CO-, C3-6 -cycloalkyl- and C 3-6 -cycloalkyl-CO-, and wherein the C 1-4 -alkyl-, C 1-4 -alkyl-O-, C 1-4 -alkyl-CO-, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, C 3-6 -cycloalkyl-, C 3-6 -cycloalkyl-CO- or C 3-6 -cycloalkyl-O-substituents may be substituted by one or more substituents independently selected from fluorine, -CF3, -CHF2, -CH2F and -CN; R 2 Selected from hydrogen, C 1-4 -alkyl-, C 1-4 -alkyl-O-, -CN and C 3-6 -cycloalkyl-, wherein the C 1-4 -alkyl-, C 1-4 -alkyl-O- and C 3-6 -cycloalkyl groups may each be optionally substituted with 1, 2, 3 or more substituents independently selected from fluorine, -CF3, -CHF2, -CH2F and -CN; R 3 Selected from C 1-6 -alkyl-O-, C 3-6 -cycloalkyl-O-, morpholino, pyrazolyl and 4 to 7 membered monocyclic heterocycloalkyl-O-, which has 1 oxygen atom as a ring member and optionally 1 or 2 independently selected from O, N and S(O) s heteroatoms, wherein s=0, 1 or 2, wherein the C 1-6 Alkyl-O- and the C 3-6 Cycloalkyl-O- may be optionally substituted with 1, 2, 3 or more substituents independently selected from fluorine, -CF3, -CHF2, -CH2F, -CN, C 1-4 -alkyl-, C 3-6 -cycloalkyl-, C 1-6 -alkyl-O- and C 3-6 -cycloalkyl-O-; R 4 is hydrogen; or R 3 and R 4 Together with the ring atoms of the phenyl group to which they are attached, they can form a 4-, 5- or 6-membered monocyclic partially saturated heterocycloalkyl or heteroaryl group, each of which has 1, 2 or 3 atoms independently selected from O, N and S(O) s wherein s=0, 1 or 2, of which in the general formula (I) there must be 1 heteroatom directly attached to R 3wherein the heterocycloalkyl group may be optionally substituted with 1, 2, 3 or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F, -CN, C 1-4 -alkyl-, C 3-6 -cycloalkyl-, C 1-6 -alkyl-O-, C 3-6 -cycloalkyl-O-, oxetanyl-O-, tetrahydrofuranyl-O- and tetrahydropyranyl-O-; R 5 is hydrogen; R 6 Selected from hydrogen, C 1-4 -alkyl-SO2-, C 3-6 -cycloalkyl-SO2 and -CN; R 7 is hydrogen; or a) R 6 and R 7 or b) R 6 and R 5 One of the pairs together with the ring atoms of the phenyl to which they are attached forms a ring having 1, 2 or 3 atoms independently selected from O, N and S(O) u 5 or 6 membered partially saturated monocyclic heterocycloalkyl containing a heteroatom, wherein u=0, 1 or 2, wherein in the general formula (I) there must be one directly connected to R 6 wherein the heterocycloalkyl group may be optionally substituted by 1, 2, 3 or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F, -CN, C 1-4 -alkyl-, C 1-6 -alkyl-O- and C 3-6 -cycloalkyl-O- or a pharmaceutically acceptable salt thereof.

[0061] In certain embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0062] In certain embodiments, the subject is a subject in need thereof.

[0063] In certain embodiments, the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, is administered in a therapeutically effective amount. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] Figure 1 Shown are Western blot determinations of ferrochelatase (FECH) protein expression levels of various K562 clones.

[0065] Figure 2 Shown are flow cytometric determinations of protoporphyrin IX (PPIX) levels of K562 clones.

[0066] Figure 3 Shown are heme and PPIX levels of WT K562 and clone 1-9 cells as determined by LC / MS / MS.

[0067] Figure 4 Shown are the effects of bitopertin and PF-03463275 on PPIX levels as determined by flow cytometry.

[0068] Figure 5 Shown are the effects of bitopertin and PF-03463275 on cell viability as measured by the Vi-CELL XR Complete System.

[0069] Figure 6 Shown are the effects of bitopertin treatment on 5-aminolevulinic acid (5-ALA) levels in clone 1-9 cells.

[0070] Figure 7 Shown are the effects of bitopertin treatment on PPIX levels in clone 1-9 cells.

[0071] Figure 8 Shown are the effects of bitopertin treatment on heme levels in clone 1-9 cells.

[0072] Figure 9 Shown are relative FECH mRNA levels in human hematopoietic stem cells after transduction with a lentiviral vector expressing shRNA for FECH.

[0073] Figure 10 Shown are flow cytometric determinations of the effects of bitopertin treatment on erythroid antigen profile and protoporphyrin IX (PPIX) levels in human hematopoietic stem cells.

[0074] Figure 11 It is shown that Biotopertin (100 nM) treatment reduced PPIX accumulation by 60%. DETAILED DESCRIPTION

[0075] Unless defined otherwise, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed embodiments belong.

[0076] As used herein, "a" or "an" means "at least one" or "one or more" unless the context clearly indicates otherwise.

[0077] As used herein, the term "about" means that the numerical value is approximate and that small variations do not significantly affect the practice of the disclosed embodiments. Where numerical limitations are used, unless the context indicates otherwise, "about" means that the numerical value can vary by ±10% and remain within the scope of the disclosed embodiments.

[0078] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0079] As used herein, the term "acylamino" refers to an amino group substituted with an acyl group (e.g., -OC(=O)-H or -OC(=O)-alkyl). Examples of acylamino groups are -NHC(=O)H or -NHC(=O)CH3. The term "lower acylamino" refers to an amino group substituted with a lower acyl group (e.g., -OC(=O)-H or -OC(=O)-C 1-6 Examples of lower acylamino groups are -NHC(=O)H or -NHC(=O)CH3.

[0080] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0081] As used herein, the term "alkenyl" refers to a straight or branched chain alkyl group having one or more carbon-carbon double bonds and 2-20 carbon atoms, including but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, etc. In some embodiments, the alkenyl chain is 2 to 10 carbon atoms in length, 2 to 8 carbon atoms in length, 2 to 6 carbon atoms in length, or 2 to 4 carbon atoms in length.

[0082] The terms "alkoxy," "phenoxy," "benzyloxy," and "pyrimidinyloxy" refer to optionally substituted alkyl, phenyl, benzyl, or pyrimidinyl groups, respectively, bonded through an oxygen atom. For example, the term "alkoxy" refers to a linear or branched -O-alkyl group of 1 to 20 carbon atoms, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, tert-butoxy, and the like. In some embodiments, the alkoxy chain is 1 to 10 carbon atoms in length, 1 to 8 carbon atoms in length, 1 to 6 carbon atoms in length, 1 to 4 carbon atoms in length, 2 to 10 carbon atoms in length, 2 to 8 carbon atoms in length, 2 to 6 carbon atoms in length, or 2 to 4 carbon atoms in length.

[0083] As used herein, the term "alkyl" refers to a straight or branched saturated hydrocarbon group. The alkyl group can contain 1 to 20, 2 to 20, 1 to 10, 2 to 10, 1 to 8, 2 to 8, 1 to 6, 2 to 6, 1 to 4, 2 to 4, 1 to 3 or 2 or 3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, tert-butyl, isobutyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, 2-methyl-1-propyl, 2-methyl-2 -propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2-methyl-1-pentyl, 2,2-dimethyl-1-propyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl and the like.

[0084] As used herein, the term "alkylamino" refers to an amino group substituted by an alkyl group having 1 to 6 carbon atoms. An example of an alkylamino group is -NHCH2CH3.

[0085] As used herein, the term "alkylene" or "alkylenyl" refers to a divalent alkyl linking group. An example of an alkylene group is methylene (-CH2-).

[0086] As used herein, the term "alkylthio" refers to an -S-alkyl group having 1 to 6 carbon atoms. An example of an alkylthio group is -SCH2CH3.

[0087] As used herein, the term "alkynyl" refers to a straight or branched chain alkyl group having one or more carbon-carbon triple bonds and 2-20 carbon atoms, including but not limited to acetylene, 1-propylene, 2-propylene, etc. In some embodiments, the alkynyl chain is 2 to 10 carbon atoms in length, 2 to 8 carbon atoms in length, 2 to 6 carbon atoms in length, or 2 to 4 carbon atoms in length.

[0088] As used herein, the term "amide" refers to a group

[0089]

[0090] Each R 30 independently represent hydrogen or a hydrocarbon group, or two R 30 Together with the nitrogen atom to which they are attached they form a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0091] As used herein, the term "amidino" refers to -C(=NH)NH2.

[0092] The terms "amine" and "amino" are art-recognized and refer to unsubstituted and substituted amines and salts thereof, for example, the moiety represented by:

[0093]

[0094] Each R 30 independently represent hydrogen or a hydrocarbon group, or two R 30 Together with the nitrogen atom to which they are attached they form a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0095] As used herein, the term "aminoalkoxy" refers to an alkoxy group substituted with an amino group. An example of an aminoalkoxy group is -OCH2CH2NH2.

[0096] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group. An example of an aminoalkyl group is -CH2CH2NH2.

[0097] As used herein, the term "aminosulfonyl" refers to -S(=O)2NH2.

[0098] As used herein, the term "aminoalkylthio" refers to an alkylthio group substituted with an amino group. An example of an aminoalkylthio group is -SCH2CH2NH2.

[0099] As used herein, the term "amphiphilic" refers to a three-dimensional structure having discrete hydrophobic and hydrophilic regions. Amphiphilic compounds suitably have both hydrophobic and hydrophilic elements present.

[0100] As used herein, the term "animal" includes, but is not limited to, humans and non-human vertebrates, such as wild animals, domestic animals, and farm animals.

[0101] As used herein, the term "aryl" refers to a monocyclic, bicyclic, or polycyclic (e.g., having 2, 3, or 4 fused rings) aromatic hydrocarbon. In some embodiments, the aryl group has 6 to 20 carbon atoms or 6 to 10 carbon atoms. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, indenyl, tetrahydronaphthyl, and the like. Examples of aryl groups include, but are not limited to:

[0102]

[0103]

[0104] As used herein, the term "arylalkyl" refers to a C 1-6 alkyl.

[0105] As used herein, the term "arylamino" refers to an amino group substituted with an aryl group. An example of an arylamino group is -NH(phenyl).

[0106] As used herein, the term "arylene" refers to an aryl linking group, ie, an aryl group that connects one group to another group in a molecule.

[0107] The term "carbamate" is art-recognized and refers to a group

[0108]

[0109] where R 29 and R 30 independently represents hydrogen or a hydrocarbon group, such as an alkyl group, or R 29 and R 30 Together with one or more intervening atoms, the heterocyclic ring comprises from 4 to 8 atoms in the ring structure.

[0110] As used herein, the term "carbamoyl" refers to -C(=O)-NH2.

[0111] As used herein, the term "carbocycle" refers to a 5- or 6-membered saturated or unsaturated ring, optionally containing an O, S, or N atom as part of the ring. Examples of carbocycles include, but are not limited to, cyclopentyl, cyclohexyl, cyclopenta-1,3-diene, phenyl, and any of the above heterocycles.

[0112] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0113] The term "carbonate" is art-recognized and refers to the group -OCO2-R 30 , where R 30 Represents a hydrocarbon group.

[0114] As used herein, the term "carboxyl" refers to a group represented by the formula CO2H.

[0115] As used herein, the term "carrier" refers to a diluent, adjuvant, or excipient with which a compound is administered. Pharmaceutical carriers can be liquids such as water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Pharmaceutical carriers can also be saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silicon dioxide, urea, and the like. In addition, adjuvants, stabilizers, thickeners, lubricants, and colorants can be used.

[0116] As used herein, the term "compound" refers to all stereoisomers, tautomers, and isotopes of the compounds described herein.

[0117] As used herein, the terms "comprising" (and any form of comprising, such as "comprise," "comprises," and "comprised"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "includes" and "include"), or "containing" (and any form of containing, such as "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0118] As used herein, the term "contacting" means bringing two elements together in an in vitro system or an in vivo system. For example, "contacting" a GlyT1 transporter inhibitor with a GlyT1 transporter and an individual or patient or cell includes administering the compound to an individual or patient, such as a human, as well as, for example, introducing the compound into a sample containing cells or a purified preparation (containing the GlyT1 transporter).

[0119] As used herein, the term "cyano" refers to -CN.

[0120] As used herein, the term "cycloalkyl" means a non-aromatic cyclic hydrocarbon, including cyclized alkyl, alkenyl and alkynyl groups containing up to 20 ring carbon atoms. Cycloalkyl can include monocyclic or polycyclic systems, such as fused ring systems, bridged ring systems and spirocyclic systems. In some embodiments, the polycyclic system comprises 2, 3 or 4 fused rings. Cycloalkyl can contain 3 to 15, 3 to 10, 3 to 8, 3 to 6, 4 to 6, 3 to 5 or 5 or 6 ring carbon atoms. The ring carbon atoms of cycloalkyl can optionally be substituted with oxo or sulfido. The example of cycloalkyl includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinenyl, norcaryl, adamantyl etc. Also included within the definition of cycloalkyl are moieties having one or more aromatic rings fused (having a common bond) to a cycloalkyl ring, such as benzo or thienyl derivatives of pentane, pentene, hexane, and the like (e.g., 2,3-dihydro-1H-inden-1-yl or 1H-inden-2(3H)-on-1-yl).

[0121] As used herein, the term "cycloalkylalkyl" refers to a C 1-6 alkyl.

[0122] As used herein, the term "dialkylamino" means an amino group substituted with two alkyl groups each having 1 to 6 carbon atoms.

[0123] As used herein, the term "diazoamino" refers to -N(NH2)2.

[0124] As used herein, the term "ester" refers to the group -C(O)OR 30 , where R 30 Represents a hydrocarbon group.

[0125] As used herein, the term "ether" refers to a hydrocarbon group attached to another hydrocarbon group through an oxygen. Thus, an ether substituent of a hydrocarbon group can be hydrocarbon-O-. Ethers can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl," which can be represented by the general formula alkyl-O-alkyl.

[0126] As used herein, the term "facially amphiphilic" or "facialamphiphilicity" refers to compounds having polar (hydrophilic) and nonpolar (hydrophobic) side chains that adopt one or more conformations that result in the separation of the polar and nonpolar side chains into opposite faces or separate regions of a structure or molecule.

[0127] As used herein, the term "glycine transporter" or "GlyT" refers to a membrane protein that facilitates the transport of glycine across the plasma membrane. Non-limiting examples of glycine transporters include glycine transporter 1 (GlyT1) and glycine transporter 2 (GlyT2).

[0128] As used herein, the term "GlyT1" or "GlyT1 transporter" means sodium- and chloride-dependent glycine transporter 1, also known as glycine transporter 1, a protein encoded by the SLC6A9 gene in humans (Kim KM, Kingsmore SF, Han H, Yang-Feng TL, Godinot N, Seldin MF, Caron MG, Giros B (June 1994). "Cloning of the human glycine transporter type 1: molecular and pharmacological characterization of novel isoform variants and chromosomal localization of the gene in the human and mouse genomes". Mol Pharmacol. 45(4):608-17; Jones EM, Fernald A, Bell GI, Le Beau MM (November 1995). "Assignment of SLC6A9 to human chromosome band 1p33 by in situ hybridization". Cytogenet Cell Genet. 71(3):211), which is hereby incorporated by reference in its entirety.

[0129] As used herein, the term "GlyT2" or "GlyT2 transporter" means sodium- and chloride-dependent glycine transporter 2, also known as glycine transporter 2, a protein encoded by the SLC6A5 gene in humans (Morrow JA, Collie IT, Dunbar DR, Walker GB, Shahid M, Hill DR (November 1998). "Molecular cloning and functional expression of the human glycine transporter GlyT2 and chromosomal localisation of the gene in the human genome". FEBS Lett. 439(3):334-40), which is hereby incorporated by reference in its entirety.

[0130] As used herein, the term "GlyT1 inhibitor" refers to a compound that inhibits or blocks the activity of the GlyT1 transporter, including compounds that inhibit the activity of any GlyT1 isoform. Non-limiting examples of GlyT1 inhibitors are provided herein. In some embodiments, the GlyT1 inhibitor is a specific GlyT1 inhibitor, meaning that the inhibitor has greater inhibitory activity against GlyT1 than against GlyT2. In some embodiments, the inhibitor selectively inhibits GlyT1 by at least or about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% compared to GlyT2. In some embodiments, the GlyT1 inhibitor inhibits GlyT1 but does not inhibit or significantly inhibits the activity of GlyT2. If the GlyT1 inhibitor inhibits the activity of GlyT2 by less than 5%, 4%, 3%, 2%, or 1%, then the GlyT1 inhibitor does not significantly inhibit the activity of GlyT2. The selectivity of GlyT1 inhibitors is determined based on assays known in the art, such as those described in published journal articles (BN Atkinson, SC Bell, M. De Vivo, LR Kowalski, SM Lechner, VIOgnyanov, C.-S. Tham, C. Tsai, J. Jia, D. Ashton and MA Klitenick, ALX 5407: A Potent, Selective Inhibitor of the hGlyT1 Glycine Transporter, Molecular Pharmacology 2001 November, 60(6): 1414-1420), which is incorporated in its entirety.

[0131] As used herein, the term "GlyT2 inhibitor" refers to a compound that inhibits or blocks the activity of the GlyT2 transporter, including compounds that inhibit the activity of any GlyT2 isoform. In some embodiments, the GlyT2 inhibitor is a nonspecific inhibitor, which means that it can also inhibit or block the activity of GlyT1. In some embodiments, the GlyT2 inhibitor is a specific GlyT2 inhibitor, which means that the inhibitor has greater inhibitory activity against GlyT2 than against GlyT1. In some embodiments, the inhibitor selectively inhibits GlyT2 by at least or about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% compared to GlyT1. In some embodiments, the GlyT2 inhibitor inhibits GlyT2 activity but does not inhibit or significantly inhibits the activity of GlyT1. A GlyT1 inhibitor does not significantly inhibit the activity of GlyT1 if the GlyT2 inhibitor inhibits the activity of GlyT1 by less than 5%, 4%, 3%, 2% or 1%. The selectivity of a GlyT2 inhibitor is determined based on assays known in the art, such as those described in a published journal article (BN Atkinson, S.C. Bell, M. De Vivo, L.R. Kowalski, S.M. Lechner, V.O. Gnyanov, C.-S. Tham, C. Tsai, J. Jia, D. Ashton and M.A. Klitenick, ALX 5407: A Potent, Selective Inhibitor of the hGlyT1 Glycine Transporter, Molecular Pharmacology 2001 November, 60(6): 1414-1420), which is incorporated herein by reference in its entirety.

[0132] As used herein, the term "guanidino" refers to -NH(=NH)NH2.

[0133] As used herein, the term "halo" refers to a halogen group including, but not limited to, fluoro, chloro, bromo, and iodo.

[0134] As used herein, the term "haloalkoxy" refers to -O-haloalkyl. An example of a haloalkoxy group is OCF3.

[0135] As used herein, the term "haloalkyl" refers to a C 1-6 Examples of haloalkyl groups include, but are not limited to, CF3, C2F5, CH2F, CHF2, CCl3, CHCl2, CH2CF3, and the like.

[0136] As used herein, the term "heteroaryl" means an aromatic heterocycle having up to 20 ring atoms (e.g., C) and having at least one heteroatom ring member (ring atoms) such as sulfur, oxygen or nitrogen. In some embodiments, heteroaryl has at least one or more heteroatom ring atoms, each of which is independently sulfur, oxygen or nitrogen. In some embodiments, heteroaryl has 3 to 20 ring atoms, 3 to 10 ring atoms, 3 to 6 ring atoms or 3 to 5 ring atoms. In some embodiments, heteroaryl contains 2 to 14 carbon atoms, 2 to 7 carbon atoms or 5 or 6 carbon atoms. In some embodiments, heteroaryl has 1 to 4 heteroatoms, 1 to 3 heteroatoms or 1 or 2 heteroatoms. Heteroaryl includes monocyclic and polycyclic (e.g., with 2, 3 or 4 condensed rings) systems. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furanyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl (such as indol-3-yl), pyrrolyl, oxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolyl, indolinyl, pyranyl, oxadiazolyl, isoxazolyl, triazolyl, thianthrenyl, pyrazolyl, indolizinyl, isoindolyl, isobenzofuranyl, benzoxazolyl, xanthenyl, 2H-pyrrolyl, pyrrolyl, 3H-indolyl, 4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinazolinyl, phenanthridinyl, acridinyl, piperidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furanyl, phenoxazinyl, and the like. Suitable heteroaryl groups include 1,2,3-triazole, 1,2,4-triazole, 5-amino-1,2,4-triazole, imidazole, oxazole, isoxazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 3-amino-1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, pyridine, and 2-aminopyridine.

[0137] As used herein, the term "heteroarylalkyl" refers to a C 1-6 alkyl.

[0138] As used herein, the term "heteroarylamino" refers to an amino group substituted by a heteroaryl group. An example of a heteroarylamino group is -NH-(2-pyridyl).

[0139] As used herein, the term "heteroarylene" means a heteroaryl linking group, ie, a heteroaryl group that connects one group to another group in a molecule.

[0140] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Exemplary heteroatoms are nitrogen, oxygen, and sulfur.

[0141] As used herein, the term "heterocycle" or "heterocyclic ring" means a 5- to 7-membered monocyclic or bicyclic or 7- to 10-membered bicyclic heterocyclic ring system, wherein any ring may be saturated or unsaturated and consists of carbon atoms and 1 to 3 heteroatoms selected from N, O, and S, and wherein the N and S heteroatoms may be optionally oxidized and the N heteroatom may be optionally quaternized, and includes any bicyclic group in which any of the above-defined heterocycles is fused to a benzene ring. Particularly useful are rings containing one oxygen or sulfur, one to three nitrogen atoms, or one oxygen or sulfur in combination with one or two nitrogen atoms. The heterocycle may be attached to any heteroatom or carbon atom, resulting in a stable structure. Examples of heterocyclic groups include, but are not limited to, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepanyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furanyl, tetrahydrofuranyl, tetrahydropyranyl, thienyl, benzothienyl, thiomorpholinyl, thiaminyl sulfoxide, thiomorpholinyl sulfone, and oxadiazolyl. Morpholino is the same as morpholinyl.

[0142] As used herein, the term "heterocycloalkyl" means a non-aromatic heterocycle having up to 20 ring atoms, including cyclized alkyl, alkenyl and alkynyl groups, wherein one or more ring carbon atoms are replaced by heteroatoms such as O, N or S atoms. Heterocycloalkyl can be monocyclic or polycyclic (e.g., fused, bridged or spirocyclic systems). In some embodiments, heterocycloalkyl has 1 to 20 carbon atoms or 3 to 20 carbon atoms. In some embodiments, heterocycloalkyl contains 3 to 14 ring atoms, 3 to 7 ring atoms or 5 or 6 ring atoms. In some embodiments, heterocycloalkyl has 1 to 4 heteroatoms, 1 to 3 heteroatoms or 1 or 2 heteroatoms. In some embodiments, heterocycloalkyl contains 0 to 3 double bonds. In some embodiments, heterocycloalkyl contains 0 to 2 triple bonds. Examples of heterocycloalkyl include, but are not limited to, morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxole, benzo-1,4-dioxane, piperidinyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, pyrazolidinyl, thiazolidinyl, imidazolidinyl, pyrrolidin-2-one-3-yl, etc. In addition, the ring-forming carbon atoms and heteroatoms of heterocycloalkyl can be optionally substituted with oxo or thio-annul groups. For example, the ring-forming S atoms can be substituted with 1 or 2 oxo groups (forming S (O) or S (O) 2). For another example, the ring-forming C atoms can be substituted with oxo groups (forming carbonyl). Also included within the definition of heterocycloalkyl are moieties having one or more aromatic rings fused to a non-aromatic heterocycle (having a common bond), including but not limited to pyridyl, thienyl, phthalimido, naphthalimido, and benzo derivatives of heterocycles such as indolene, isoindolene, 4,5,6,7-tetrahydrothieno[2,3-c]pyridin-5-yl, 5,6-dihydrothieno[2,3-c]pyridin-7(4H)-on-5-yl, isoindolin-1-on-3-yl, and 3,4-dihydroisoquinolin-1(2H)-on-3-yl. The ring carbon atoms and heteroatoms of the heterocycloalkyl may be optionally substituted with oxo or thioxanthio.

[0143] As used herein, the term "heterocycloalkylalkyl" refers to a C 1-6 alkyl.

[0144] As used herein, the term "hydroxy" or "hydroxyl" refers to an -OH group.

[0145] As used herein, the term "hydroxyalkyl" or "hydroxylalkyl" refers to an alkyl group substituted with a hydroxy group. Examples of hydroxyalkyl groups include, but are not limited to, -CH2OH and -CH2CH2OH.

[0146] As used herein, the terms "subject" or "patient," which are used interchangeably, mean any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates such as humans.

[0147] As used herein, the phrase "inhibit an activity" such as enzyme activity or transporter activity means reducing the activity of the enzyme or transporter (such as GlyT1 transporter) by any measurable amount.

[0148] As used herein, the phrase "in need thereof" means that an animal or mammal has been identified as being in need of a particular method or treatment. In some embodiments, identification can be performed by any diagnostic means. An animal or mammal may be in need of any of the methods and treatments described herein. In some embodiments, the animal or mammal is in an environment where a particular disease, disorder, or condition is prevalent or is about to be transported to such an environment.

[0149] As used herein, the phrase "in situ gelable" is meant to include not only low viscosity liquids that form gels upon contact with the eye or tear fluid outside the eye, but also more viscous liquids such as semifluids and thixotropic gels that exhibit a significant increase in viscosity or gel stiffness when applied to the eye.

[0150] As used herein, the phrase "an integer from X to Y" means any integer including the endpoints. For example, the phrase "an integer from X to Y" means 1, 2, 3, 4, or 5.

[0151] When used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy, the term "lower" is intended to include groups wherein there are ten or fewer, preferably six or fewer, non-hydrogen atoms in the substituent. For example, "lower alkyl" refers to an alkyl group containing 10 or fewer, preferably 6 or fewer carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl or lower alkoxy, whether they occur alone or in combination with other substituents, such as the hydroxyalkyl and aralkyl groups listed (in this case, for example, when counting the carbon atoms in the alkyl substituent, the atoms in the aryl group are not counted).

[0152] As used herein, the term "mammal" means a rodent (ie, mouse, rat, or guinea pig), monkey, cat, dog, cow, horse, pig, or human. In some embodiments, the mammal is a human.

[0153] As used herein, the term "N-alkyl" refers to an alkyl chain substituted with an amine group. Non-limiting examples include, but are not limited to The alkyl chain can be linear, branched, cyclic, or any combination thereof. In some embodiments, the alkyl group contains 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2 carbons.

[0154] As used herein, the term "nitro" refers to -NO2.

[0155] As used herein, the term "n-membered" (where n is an integer) typically describes the number of ring atoms in a moiety, where the number of ring atoms is n. For example, pyridine is an example of a 6-membered heteroaryl ring and thiophene is an example of a 5-membered heteroaryl ring.

[0156] As used herein, the phrase "ophthalmologically acceptable" means having no persistent detrimental effect on the treated eye or its function, or on the general health of the treated subject. However, it should be recognized that transient effects such as mild irritation or a "stinging" sensation are common for topical ophthalmic administration of drugs, and the presence of such transient effects does not conflict with the composition, formulation, or ingredient (e.g., excipient) in question being "ophthalmologically acceptable" as defined herein.

[0157] As used herein, the phrase "optionally substituted" means that substitution is optional and, therefore, includes both unsubstituted and substituted atoms and moieties. A "substituted" atom or moiety means that any hydrogen on the designated atom or moiety can be replaced by a substituent selected from the designated substituent, provided that the normal valence of the designated atom or moiety is not exceeded and that the substitution results in a stable compound. For example, if a methyl group is optionally substituted, then three hydrogen atoms on the carbon atom can be replaced by substituents.

[0158] As used herein, the phrase "pharmaceutically acceptable" means those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals. In some embodiments, the term "pharmaceutically acceptable" means approved by a regulatory agency of the federal or state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0159] "Pharmaceutically acceptable salts" are intended to mean salts of the free acids or bases of the compounds represented herein that are non-toxic, biologically tolerable, or otherwise biologically suitable for administration to a subject. See generally SM Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977, 66, 1-19. Preferred pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of a subject without undue toxicity, irritation, or allergic response. The compounds described herein may have a sufficiently acidic group, a sufficiently basic group, both types of functional groups, or more than one of each type, and thus react with a variety of inorganic or organic bases, and inorganic and organic acids, to form pharmaceutically acceptable salts.

[0160] For compounds described herein that contain a basic group such as an amine, pharmaceutically acceptable salts can be prepared by any suitable method available in the art, for example, by treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric acid, phosphoric acid, or the like; or an organic acid such as acetic acid, phenylacetic acid, propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid, isethionic acid, succinic acid, valeric acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, oleic acid, palmitic acid, lauric acid, or the like. Acids, pyranosidic acids (such as glucuronic acid or galacturonic acid), α-hydroxy acids (such as mandelic acid, citric acid or tartaric acid), amino acids (such as aspartic acid or glutamic acid), aromatic acids (such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid or cinnamic acid), sulfonic acids (such as laurylsulfonic acid, p-toluenesulfonic acid, methanesulfonic acid or ethanesulfonic acid); or any compatible acid mixtures, such as those given as examples herein; and any other acids and mixtures thereof that are considered equivalents or acceptable substitutes according to the ordinary skill in the art.

[0161] For compounds described herein containing acidic groups (such as carboxylic acid groups), base addition salts can be prepared by any suitable method available in the art, for example, by treating such compounds with a sufficient amount of the desired base (either neat or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include, but are not limited to, lithium, sodium, potassium, calcium, ammonium, zinc or magnesium salts, or other metal salts; organic amino salts, such as alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts.

[0162] Other examples of pharmaceutically acceptable salts include, but are not limited to, camphorsulfonate, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, octanoate, acrylate, formate, isobutyrate, hexanoate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, Pharmaceutically acceptable salts include, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, benzenesulfonate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, and mandelate. A list of other suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pennsylvania, 1985.

[0163] The neutral form of the compound is preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties (such as solubility in polar solvents), but otherwise the salts are equivalent to the parent form of the compound for the purposes of this application.

[0164] As used herein, the term "phenyl" refers to -C6H5. A phenyl group may be unsubstituted or substituted with one, two or three suitable substituents.

[0165] The terms "polycyclic group," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings." Each ring of the polycyclic ring can be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring contains 3 to 10 atoms in the ring, preferably 5 to 7 atoms.

[0166] As used herein, the term "prodrug" means a derivative of a known direct-acting drug that has enhanced delivery characteristics and therapeutic value compared to the drug and is converted into an active drug by enzymatic or chemical methods. A common method for preparing a prodrug is to include one or more selected moieties that hydrolyze under physiological conditions to produce the desired molecule. In certain embodiments, the prodrug is converted by the enzymatic activity of the host animal. For example, a prodrug with a nitro group on an aromatic ring can be reduced by a reductase to produce the desired amino group of the corresponding active compound in vivo. In another example, a functional group such as a hydroxyl, carbonate, or carboxylic acid in the parent compound is presented as an ester, which can be cleaved by an esterase. In addition, the amine group in the parent compound is presented as, but not limited to, a carbamate, N-alkylated, or N-acylated form (Simplício et al., "Prodrugs for Amines," Molecules, (2008), 13: 519-547). In certain embodiments, some or all of the compounds described herein in the formulations represented above can be replaced with corresponding suitable prodrugs.

[0167] As used herein, the term "purified" means that when isolated, the isolate contains at least 90%, at least 95%, at least 98%, or at least 99% of a compound described herein by weight of the isolate.

[0168] As used herein, the phrase "quaternary ammonium salt" means a derivative of the disclosed compounds having one or more tertiary amine moieties, wherein at least one tertiary amine moiety in the parent compound is converted to a quaternary ammonium salt by alkylation (and the cation is converted to a quaternary ammonium salt by an anion such as Cl). - 、CH3COO - and CF3COO - The tertiary amine moiety is modified by converting it to a quaternary ammonium cation (e.g., methylation or ethylation).

[0169] As used herein, the term "semicarbazone" means =NNHC(=O)NH2.

[0170] As used herein, the phrase "solubilizer" means an agent that results in the formation of a micellar solution or a true solution of the drug.

[0171] As used herein, the term "solution / suspension" means a liquid composition wherein a first portion of the active agent is present in solution and a second portion of the active agent is present in suspension in the form of particles in a liquid matrix.

[0172] As used herein, the phrase "substantially isolated" means a compound that is at least partially or substantially separated from the environment in which it was formed or detected.

[0173] The term "substituted" refers to a portion of a substituent having a hydrogen on one or more carbons that replaces the main chain. It should be understood that "substituted" or "substituted by" includes implicit conditions, i.e., such substitution is consistent with the allowed valence of the substituted atom and the substituent, and that the substitution produces a stable compound, for example, the stable compound will not spontaneously transform such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all allowed substituents of an organic compound. In a broad sense, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. For appropriate organic compounds, allowed substituents may be one or more and be the same or different. For the purposes of this application, heteroatoms such as nitrogen may have hydrogen substituents and / or any allowed substituents of an organic compound described herein that satisfy the heteroatom valence.

[0174] Substituents may include any substituent described herein, such as halogen, hydroxy, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that substituents themselves may be substituted, if appropriate. Unless specifically indicated as "unsubstituted," reference to chemical moieties herein should be understood to include substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0175] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0176] The term "sulfonamide" is art-recognized and refers to a group represented by the following general formula:

[0177]

[0178] where R 29 and R 30 independently represents hydrogen or a hydrocarbon group, such as an alkyl group, or R 29 and R 30 Together with one or more intervening atoms, the heterocyclic ring comprises from 4 to 8 atoms in the ring structure.

[0179] The term "sulfoxide" is art-recognized and refers to the group -S(O)-R 30 , where R 30 Represents a hydrocarbon group.

[0180] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0181] The term "sulfone" is art-recognized and refers to the group -S(O)2-R 30 , where R 30 Represents a hydrocarbon group.

[0182] As used herein, the phrase "therapeutically effective amount" means the amount of an active compound or agent that elicits a biological or medical response in a tissue, system, animal, individual, or human being that is being sought by a researcher, veterinarian, physician, or other clinician. The therapeutic effect depends on the disorder being treated or the desired biological effect. Thus, the therapeutic effect can be a reduction in the severity of the symptoms associated with the disorder and / or an inhibition (partial or complete) of the disorder's progression, or the treatment, cure, prevention, or elimination of an improvement in the disorder or side effect. The amount required to elicit a therapeutic response can be determined based on the subject's age, health status, body shape, and sex. The optimal amount can also be determined based on monitoring the subject's response to treatment.

[0183] As used herein, the term "alkylthio" refers to an alkyl group substituted with a thiol group.

[0184] As used herein, the term "thioester" refers to the group -C(O)SR 30 or -SC(O)R 30 , where R 30 Represents a hydrocarbon group.

[0185] As used herein, the term "thioether" is equivalent to an ether in which the oxygen is replaced by sulfur.

[0186] As used herein, the term "treat, treated or treating" means therapeutic treatment and preventive measures, wherein the purpose is to slow down (mitigate) an undesirable physiological condition, disorder or disease, or to obtain a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to, relief of symptoms; alleviation of the extent of the condition, disorder or disease; a stable (i.e., non-worsening) state of the condition, disorder or disease; a delay or slowing of the onset of the condition, disorder or disease progression; an improvement or alleviation (whether partial or complete) of a detectable or undetectable condition, disorder or disease state; an improvement in at least one measurable physical parameter, which is not necessarily discernible by the patient; or an enhancement or improvement of the condition, disorder or disease. Treatment includes causing a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival time compared to the expected survival time if not receiving treatment. Thus, "treatment of erythropoietic protoporphyria" or "treating erythropoietic protoporphyria" means an activity that alleviates or ameliorates any primary phenomenon or secondary symptoms associated with erythropoietic protoporphyria or other conditions described herein.

[0187] The term "urea" is art-recognized and can be represented by the general formula

[0188]

[0189] where R 29 and R 30 independently represents hydrogen or a hydrocarbon group, such as an alkyl group, or any occurrence of R 29 With R 30 Together with one or more intervening atoms, it forms a heterocycle having 4 to 8 atoms in the ring structure.

[0190] At various places in this specification, substituents of compounds may be disclosed in groups or ranges. In particular, it is intended that the embodiments include every individual subcombination of the members of such groups and ranges. For example, the term "C 1-6 "Alkyl" is specifically intended to individually disclose methyl, ethyl, propyl, C4 alkyl, C5 alkyl, and C6 alkyl.

[0191] For compounds in which a variable occurs more than once, each variable may be a different part selected from the Markush group defining the variable. For example, where a structure is described as having two R groups present simultaneously on the same compound, the two R groups may represent different parts selected from the Markush group defined for R. In another example, when a structure is described as having two R groups present simultaneously on the same compound, the two R groups may represent different parts selected from the Markush group defined for R. When the form specifies optional multiple substituents, it is understood that the substituent R can appear s times on the ring and R can be a different moiety at each occurrence. 1 is defined to include hydrogen, such as when T 1 When it is CH2, NH, etc., any H may be replaced by a substituent.

[0192] It will also be appreciated that certain features described herein, which for clarity are described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features, which for brevity are described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

[0193] It should be understood that embodiments of the present invention include stereoisomers, diastereomers and optical stereoisomers and mixtures thereof of compounds used where applicable. Additionally, it should be understood that stereoisomers, diastereomers and optical stereoisomers and mixtures thereof of compounds are within the scope of the embodiments. As non-limiting examples, a mixture can be a racemate or a mixture can contain an unequal proportion of a specific stereoisomer relative to another. Additionally, compounds can be provided as substantially pure stereoisomers, diastereomers and optical stereoisomers (such as epimers).

[0194] The compounds described herein can be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers (such as enantiomers and diastereomers) are intended to be included within the scope of the embodiment. Compounds containing asymmetrically substituted carbon atoms can be separated in optically active or racemic form. Methods for preparing optically active forms from optically active starting materials are known in the art, such as by splitting racemic mixtures or preparing by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, etc. can also be present in the compounds described herein, and all such stable isomers are provided herein. The cis and trans geometric isomers of the compound are also included in this embodiment, and they can be separated as a mixture of isomers or as separate isomeric forms. When specifying a compound capable of stereoisomerization or geometric isomerization in its structure or name without mentioning a specific R / S or cis / trans configuration, all such isomers are intended to be covered.

[0195] In some embodiments, the composition comprises a compound or a pharmaceutically acceptable salt, solvate, or prodrug thereof that is at least 90%, at least 95%, at least 98%, or at least 99%, or 100% enantiomerically pure, meaning that the ratio of one enantiomer to the other in the composition is at least 90:1, at least 95:1, at least 98:1, or at least 99:1, or that one enantiomer is present entirely in excess of the other. In certain embodiments, a compound enriched in one enantiomer is substantially free of the other enantiomer, where "substantially free" means, for example, that the substance in question comprises less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of the amount of the other enantiomer in the composition or compound mixture. For example, if a composition or compound mixture contains 98 grams of a first enantiomer and 2 grams of a second enantiomer, it would be said to contain 98 mol% of the first enantiomer and only 2% of the second enantiomer.

[0196] In certain embodiments, a compound enriched in one enantiomer is substantially free of the other enantiomer, where substantially free means, for example, that the substance in question comprises less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% of the amount of the other enantiomer in the composition or compound mixture. For example, if a composition or compound mixture contains 98 grams of a first enantiomer and 2 grams of a second enantiomer, it would be said to contain 98 mol% of the first enantiomer and only 2% of the second enantiomer.

[0197] The splitting of the racemic mixture of compound can be carried out by any of a variety of methods known in the art, including, for example, chiral HPLC, using the fractional recrystallization of chiral resolving acid, which is a kind of optically active salified organic acid. Suitable resolving agents for fractional recrystallization method include but are not limited to optically active acid, such as tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid and various optically active camphorsulfonic acids, such as β-camphorsulfonic acid, for example. Other resolving agents suitable for fractional crystallization method include but are not limited to α-methylbenzylamine (for example, S and R form or diastereoisomerically pure form), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane etc. The splitting of racemic mixture can also be carried out by eluting on a column filled with an optically active resolving agent (for example, dinitrobenzoylphenylglycine). Suitable elution solvent composition can be determined by those skilled in the art.

[0198] Compounds can also include tautomeric forms. Tautomeric forms are produced by the exchange of a single bond with an adjacent double bond and the concomitant migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include, but are not limited to, keto-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions of a heterocyclic ring system, including but not limited to 1H- and 3H-imidazoles, 1H-, 2H-, and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

[0199] Glycine transporter inhibitors, such as GlyT1 inhibitors, including pharmaceutically acceptable salts thereof (e.g., GlyT1 inhibitors as disclosed herein) can also exist as hydrates and solvates, as well as anhydrous and non-solvated forms. A "hydrate" is a compound present in a composition together with water molecules. The composition can contain a stoichiometric amount of water, such as a monohydrate or a dihydrate, or can contain a random amount of water. A "solvate" is a similar composition, except that a solvent other than water is used, such as methanol, ethanol, dimethylformamide, diethyl ether, etc., instead of water. For example, methanol or ethanol can form an "alcoholate," which can also be stoichiometric or non-stoichiometric. Mixtures of such solvates or hydrates can also be prepared. The source of such solvates or hydrates can be from the crystallization solvent, inherent in the preparation or crystallization solvent, or foreign to such solvent.

[0200] The compounds of the present invention, including pharmaceutically acceptable salts and prodrugs thereof, can exist in various polymorphs, pseudopolymorphs or in an amorphous state. As used herein, the term "polymorph" refers to different crystalline forms and other solid molecular forms of the same compound, including pseudopolymorphs, such as hydrates, solvates or salts of the same compound. Due to changes in temperature, pressure or changes in the crystallization process, different crystalline polymorphs have different crystal structures. The physical properties of polymorphs differ from each other, such as X-ray diffraction characteristics, stability, melting point, solubility or dissolution rate in certain solvents. Therefore, crystalline polymorphs are an important aspect of developing suitable dosage forms in the pharmaceutical industry.

[0201] Compounds can also include all isotopes of atoms present in the intermediates or final compound. Isotopes include those atoms that have the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0202] In some embodiments, the compound or its salt is substantially isolated. Partial isolation can include, for example, a composition enriched in the compound. Substantial isolation can include a composition containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% of the compound or its salt by weight. Methods for isolating compounds and their salts are conventional in the art.

[0203] Although the disclosed compounds are suitable, other functional groups can be introduced into the compounds in the hope of obtaining similar results. In particular, it is expected that thioamides and thioesters have very similar properties. The distance between the aromatic rings may affect the geometric pattern of the compound, and this distance can be changed by incorporating aliphatic chains of different lengths, which can optionally be substituted or can contain amino acids, dicarboxylic acids or diamines. The distance and relative orientation between monomers in the compound can also be changed by replacing the amide bond with a substitute with an extra atom. Therefore, replacing the carbonyl with a dicarbonyl group changes the distance between the monomers and the tendency of the dicarbonyl unit to adopt the anti-arrangement of the two carbonyl parts and change the periodicity of the compound. Pyromellitic anhydride represents another alternative to simple amide bonds, which can change the conformation and physical properties of the compound. Modern methods of solid phase organic chemistry (E.Atherton and RCSheppard, Solid Phase Peptide Synthesis A Practical Approach IRL Press Oxford 1989) now allow the synthesis of uniformly dispersed compounds with molecular weights close to 5,000 daltons. Other substitution patterns are equally effective.

[0204] The compounds also include derivatives known as prodrugs.

[0205] Compounds containing amine functional groups can also form N-oxides. Compounds containing amine functional groups mentioned herein also include N-oxides. When a compound contains several amine functional groups, one or more nitrogen atoms can be oxidized to form N-oxides. Examples of N-oxides include N-oxides of nitrogen atoms of tertiary amines or nitrogen-containing heterocycles. N-oxides can be formed by treating the corresponding amine with an oxidizing agent such as hydrogen peroxide or a peracid (e.g., peroxycarboxylic acid) (See, Advanced Organic Chemistry, Jerry March, 4th edition, Wiley Interscience).

[0206] By hereby retaining the right to limit or exclude any individual member of any such group that can be claimed according to scope or in any similar manner (including any sub-range or combination of sub-ranges in the group), for any reason, less than the full scope of the present disclosure can be claimed. In addition, by hereby retaining the right to limit or exclude any individual substituent, analog, compound, ligand, structure or its group or any member of the group that claims protection, for any reason, less than the full scope of the present disclosure can be claimed. Throughout this disclosure, multiple patents, patent applications and publications are mentioned. The disclosures of these patents, patent applications and publications are incorporated into this disclosure in their entirety by reference, to more fully describe the prior art known to those skilled in the art as of the date of this disclosure. In the event of any inconsistency between the cited patents, patent applications and publications and this disclosure, this disclosure will be the standard.

[0207] For convenience, certain terms employed in the specification, examples, and claims are collected here. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0208] Various embodiments of compounds and their salts are provided. Where a variable is not specifically listed, the variable can be any of the options described herein unless otherwise stated or indicated by the context.

[0209] In some embodiments, the compound is as described in the accompanying exemplary non-limiting claims, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0210] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0211]

[0212] in:

[0213] Ar is unsubstituted or substituted aryl or 6-membered heteroaryl containing 1, 2 or 3 nitrogen atoms, wherein the substituted aryl and the substituted heteroaryl are substituted by one or more substituents selected from the group consisting of hydroxy, halogen, NO2, CN, (C1-C6)-alkyl, (C1-C6)-alkyl substituted by halogen, (C1-C6)-alkyl substituted by hydroxy, (CH2)n-(C1-C6)-alkoxy, (C1-C6)-alkoxy substituted by halogen, NR 7 R 8 、C(O)R 9 、SO2R 10 and -C(CH3)=NOR 7, or substituted by a 5-membered aromatic heterocycle containing 1 to 4 heteroatoms selected from N and O, said aromatic heterocycle being optionally substituted by a (C1-C6)-alkyl group;

[0214] R 1 is hydrogen or (C1-C6)-alkyl;

[0215] R 2 is hydrogen, (C1-C6)-alkyl, (C2-C6)-alkenyl, (C1-C6)-alkyl substituted by halogen, (C1-C6)-alkyl substituted by hydroxy, (CH2)n-(C3-C7)-cycloalkyl, CH(CH3)-(C3-C7)-cycloalkyl, (CH2) n+1 -C(O)-R 9 、(CH2) n+1 -CN, bicyclo[2.2.1]heptyl, (CH2) n+1 -O-(C1-C6)-alkyl, (CH2) n -heterocycloalkyl, (CH2) n -aryl or (CH2) containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur or nitrogen n -5- or 6-membered heteroaryl, wherein aryl, heterocycloalkyl and heteroaryl are unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, (C1-C6)-alkyl and (C1-C6)-alkoxy;

[0216] R 3 、R 4 and R 6 are each independently hydrogen, hydroxy, halogen, (C1-C6)-alkyl, (C1-C6)-alkoxy or O-(C3-C6)-cycloalkyl;

[0217] R 5 NO2, CN, C(O)R 9 or SO2R 10 ;

[0218] R 7 and R 8 are each independently hydrogen or (C1-C6)-alkyl;

[0219] R 9 is hydrogen, (C1-C6)-alkyl, (C1-C6)-alkoxy or NR 7 R 8 ;

[0220] R 10 is (C1-C6)-alkyl, (CH2) optionally substituted by halogen n -(C3-C6)-cycloalkyl, (CH2)n -(C3-C6)-alkoxy, (CH2) n -heterocycloalkyl or NR 7 R 8 ;

[0221] n is 0, 1, or 2;

[0222] or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0223] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a GlyT1 inhibitor having the formula The compound bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0224] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0225]

[0226] in:

[0227] R1 represents a heteroaryl group selected from the group consisting of imidazolyl, thiazolyl, pyridinyl, oxazolyl, pyrazolyl, triazolyl, oxadiazolyl, quinolinyl, isoxazolyl, pyrroloimidazolyl, and thiadiazole, wherein the heteroaryl group is optionally substituted with one or more substituents selected from the group consisting of -OH, -NR7R8, halogen, (C1-C8)alkyl, (C3-C 10 )cycloalkyl, (C1-C8)alkoxy, (C1-C 12 ) alkoxyalkyl, (C1-C8) hydroxyalkyl, (C6-C 14 ) aryl and benzyl;

[0228] R2, R3 and A independently represent H or (C1-C8)alkoxy, wherein the alkyl group is optionally substituted with one or more -OH, (C1-C8)alkoxy, -NR7R8 or halogen;

[0229] Q represents -(CH2) n -, wherein n=1, 2, 3 or 4, or -(CH2) m -O-, wherein m=2, 3 or 4;

[0230] Z represents (C6-C 14 )aryl, (C1-C8)alkyl or (C3-C8)cycloalkyl;

[0231] R4 and R5 each independently represent H, halogen, (C1-C8) alkyl, (C6-C 14 )aryl, (C6-C14 )aryloxy, (C1-C8)alkoxy, (3-10 membered)heterocycloalkyl or (C3-C8)cycloalkoxy; wherein R4 and R5 are optionally substituted with one or more -OH, (C1-C8)alkoxy, -NR7R8 or halogen;

[0232] Y represents -R6, -(CH2)o-R6, -C(R6)3 or -CH(R6)2, where 0=1, 2 or 3;

[0233] R6 represents H, (C6-C 14 ) aryl, (C 1-10 )alkyl, (C3-C 10 )cycloalkyl, (C5-C 18 )bicycloalkyl, (C5-C 18 )tricycloalkyl, (3-10 membered)heterocycloalkyl, (5-10 membered)heteroaryl, -C(=O)NR7R8 or -C(=O)OR7, wherein the R6 group may be optionally substituted with one or more X groups;

[0234] Where X=-OH, (C1-C8) alkoxy, -NR 11 R 12 、-SO2R 10 、-C(=O)R 10 , halogen, cyano, (C1-C8) alkyl, (C1-C 10 )alkoxyalkyl, (5-10 membered)heteroaryl, (C6-C 14 )aryl, (C6-C 14 )aryloxy, benzyl or (C1-C8)hydroxyalkyl;

[0235] wherein R7 and R8 independently represent H, (C1-C8)alkyl, (C3-C8)cycloalkyl, (5-10 membered)heterocycloalkyl, (C1-C8)hydroxyalkyl, (5-10 membered)heteroaryl or (C1-C8) 10 ) alkoxyalkyl; wherein R7 and R8 may be optionally substituted with one or more X groups;

[0236] or R7 and R8 together with the nitrogen to which they may be attached may form a (3-10 membered) heterocycloalkyl optionally substituted with one or more X groups;

[0237] where R 10 represents (C1-C8)alkyl, (C3-C8)cycloalkyl, (3-10 membered)heterocycloalkyl, (C1-C8)hydroxyalkyl, (5-10 membered)heteroaryl or (C1-C 10 ) alkoxyalkyl;

[0238] where R 11 and R12 independently represents H, (C1-C8)alkyl, (C3-C8)cycloalkyl, (5-10 membered)heterocycloalkyl, (C1-C8)hydroxyalkyl, (5-10 membered)heteroaryl or (C1-C8) 10 )alkoxyalkyl; or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0239] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0240] or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0241] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0242] PF-3463275, or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0243] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0244]

[0245] in:

[0246] Z 1 Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, halo C 1-4 Alkyl, phenyl, halogenated C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkylsulfonyloxy, C 1-4 alkylsulfonyl, bromine, and chlorine;

[0247] Z 2 Selected from hydrogen, halogen, cyano, C 1-4 Alkyl, phenyl, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkoxy C 1-4 Alkyl and C 3-6 Cycloalkyl;

[0248] Z 3 Selected from hydrogen, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C1-4 Alkylthio, halo C 1-4 Alkyl, halogenated C 1-4 Alkoxy and C 3-6 Cycloalkyl;

[0249] Z 4 Selected from hydrogen, halogen, C1-3 alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, phenyl, halo C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkoxy C 1-4 Alkyl and C 3-6 Cycloalkyl;

[0250] Z 5 Selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, phenyl, halo C 1-4 Alkyl, halogenated C 1-4 Alkoxy, halogenated phenyl, C 1-4 Alkoxy C 1-4 Alkyl and C 3-6 Cycloalkyl;

[0251] If Z 1 to Z 5 If more than one of them is a methoxy group, then only Z 1 and Z 5 is methoxy, R 3 and R 4 independently selected from hydrogen and C optionally substituted by one or more groups Y 1-4 Alkyl; or R 3 and R4 together with the nitrogen atom to which they are attached form a saturated or partially unsaturated A, 5-, 6- or 7-membered carbocyclic ring optionally substituted with a group Y';

[0252] Y is selected from C 1-4 Alkoxy, hydroxy, halogenated C 1-4 Alkoxy and C 3-5 Cycloalkyl;

[0253] Y' is selected from C 1-4 Alkyl, C 1-4 Alkoxy, halogen, hydroxyl, halogenated C 1-4 Alkoxy, C 3-5 Cycloalkyl and C 5-10 Aryl, or Y' forms a -CH2- or -CH2-CH2- bridge between two atoms on A, a 5-, 6-, or 7-membered carbocyclic ring;

[0254] R 5and R 6 is independently C optionally substituted with one or more groups X 1-4 Alkyl; or R 5 and R 6 Together with the carbon atoms to which they are attached, they form a saturated 5- or 6-membered carbocyclic ring optionally substituted by one or more groups X', in which R 5 In case R6 and R6 together with the carbon atom to which they are attached form a 5-membered saturated carbocyclic ring, the ring may optionally further comprise an additional heteroatom group selected from O, N and S(O)m; wherein m=0, 1 or 2.

[0255] X is selected from halogen, hydroxyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy and C 5-10 Aryl; and X' is selected from halogen, hydroxy, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy and C 5-10 aryl;

[0256] where R 3 、R 4 、R 5 and R 6 Not all are unsubstituted methyl groups;

[0257] The condition is that when Z 1 is a propoxy group, Z 3 It is chlorine, Z 2 =Z 4 =Z 5 =H, and R 5 and R 6 When both are methyl groups, then R 3 and R 4 Together with the nitrogen atom to which they are attached, they do not form a 2-methylpyrrolidino group; when Z 1 is methyl, Z 3 is methoxy, Z 2 =Z4=Z5=H, and R 5 and R 6 When both are methyl groups, then R 3 and R 4 Together with the nitrogen atom to which they are attached, they do not form a pyrrolidine group, or a pharmaceutically acceptable salt thereof, or a prodrug of said compound or a pharmaceutically acceptable salt thereof.

[0258] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0259] a compound or a pharmaceutically acceptable salt thereof or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0260] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the following formula

[0261]

[0262] wherein:

[0263] Z is (CH2) n , O, S, SO, SO2 or N-R5;

[0264] n is 0, 1 or 2;

[0265] X represents 1-3 substituents independently selected from: hydrogen, halogen,

[0266] (C 1-6 )alkoxy, (C 3-6 )cycloalkoxy, (C 6-12 )aryloxy, (C 6-12 )aryl, thienyl, SR6, SOR6,

[0267] SO2R6, NR6R6, NHR6, NH2, NHCOR6, NSO2R6, CN, COOR6 and (C 1-4 )alkyl, which is optionally substituted by halogen, (C 6-12 )aryl, (C 1-6 )alkoxy or (C 6-12 )aryloxy; or two substituents in adjacent positions together represent a fused (C 5-6 )aryl, a fused (C 5-6 )cycloalkyl ring or O-(CH2) m -O; m is 1 or 2;

[0268] Y represents 1-3 substituents independently selected from hydrogen, halogen, (C 1-4 )alkoxy, SR6, NR6R6 and (C 1-4 )alkyl, which is optionally substituted by halogen;

[0269] R1 is COOR7 or CONR8R9;

[0270] R2 and R6 are (C 1-4 )alkyl;

[0271] R3, R4 and R5 are independently hydrogen or (C 1-4 )alkyl;

[0272] R7, R8 and R9 are independently hydrogen, (C 1-4)alkyl, (C 6-12 ) aryl or aralkyl, or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0273] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0274] ORG-25935, or a pharmaceutically acceptable salt thereof, or a prodrug of said compound or a pharmaceutically acceptable salt thereof.

[0275] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0276]

[0277] in:

[0278] n is an integer from 1 to 3;

[0279] R 1 and R 2 R is independently selected from hydrogen, alkyl, haloalkyl, alkoxy, haloalkoxy, aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein the above ring is optionally independently selected from the following R a 、R b or R c Substituted: alkyl, halogen, haloalkyl, alkoxy, haloalkoxy, hydroxy, cyano, monosubstituted amino or disubstituted amino; or R 1 and R 2 When attached to the same carbon atom, they may combine to form a cycloalkyl or monocyclic saturated heterocyclic group to give a spirocycle, wherein the cycloalkyl or monocyclic saturated heterocyclic group may optionally be independently selected from the following R d 、R c or R f Substituted: alkyl, alkoxy, fluorine, fluoroalkyl, fluoroalkoxy, hydroxy, monosubstituted amino or disubstituted amino; or R 1 and R 2 when attached to carbon atoms at positions 2 and 5 or 3 and 6 of the piperazine ring can combine to form a -C1-C3-alkylene chain, wherein one carbon atom in the alkylene chain is optionally replaced by -NR-, -O-, -S(O)n- (wherein R is hydrogen or alkyl and n is 0-2), and further wherein one or two hydrogen atoms in the alkylene chain can be optionally substituted by one or two alkyl groups;

[0280] R 3 、R 4 and R 5 are independently hydrogen, alkyl, fluorine or fluoroalkyl; and Ar1 and Ar 2 are independently aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein each of the above rings is optionally replaced by R g 、R h or Ri substituted, where R g is an alkyl group, -C=CR 6 (where R 6 is aryl or heteroaryl), halogen, haloalkyl, haloalkoxy, alkylthio, cyano, alkoxy, amino, monosubstituted amino, disubstituted amino, sulfonyl, acyl, carboxyl, alkoxycarbonyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, aminosulfonyl, aminocarbonyl or acylamino, and R h and R i are independently selected from alkyl, halogen, haloalkyl, haloalkoxy, alkylthio, cyano, alkoxy, amino, monosubstituted amino, disubstituted amino, sulfonyl, acyl, carboxyl, alkoxycarbonyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, aminosulfonyl, aminocarbonyl, acylamino, aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein R g 、R h and R i The aromatic or alicyclic ring in is optionally replaced by R j 、R k or R l substituted, said Rj, Rk or Rl are independently selected from alkyl, halogen, haloalkyl, haloalkoxy, alkylthio, cyano, alkoxy, amino, monosubstituted amino, disubstituted amino, sulfonyl, acyl, carboxyl, alkoxycarbonyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, aminosulfonyl, aminocarbonyl or acylamino; or a pharmaceutically acceptable salt thereof, provided that: the compound of formula V is not 2-(4-diphenylmethylpiperazin-1-yl)acetic acid, 2-(4-( (4-chlorophenyl)(phenyl)methyl)piperazin-1-yl)acetic acid, 2-((2R,5S)-4-((R)-(4-(lH-tetrazol-5-yl)phenyl)(3-hydroxyphenyl)methyl)-2,5-dimethylpiperazin-1-yl)acetic acid, or 2-((2R,5S)-4-((R)-(4-cyanophenyl)(3-hydroxyphenyl)methyl)-2,5-dimethylpiperazin-1-yl)acetic acid, or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0281] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0282] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0283] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0284]

[0285] in:

[0286] A represents a group of the general formula N-R1, a group of the general formula N+(O-)R1 or a group of the general formula N+(R')R1, and wherein R1 represents a hydrogen atom, or a straight-chain or branched (C1-C7)alkyl group optionally substituted by one or more fluorine atoms, or a (C4-C7)cycloalkyl group, or a (C3-C7)cycloalkyl (C1-C3)alkyl group, or a phenyl (C1-C3)alkyl group optionally substituted by one or two hydroxyl or methoxy groups, or a (C2-C4)alkenyl group, or a (C2-C4)alkynyl group,

[0287] R' represents a linear or branched (C1-C7) alkyl group,

[0288] X represents a hydrogen atom or one or more substituents selected from a halogen atom and a trifluoromethyl group, a linear or branched (C1-C4) alkyl group and a (C1-C4) alkoxy group;

[0289] R2 represents a hydrogen atom, or one or more substituents selected from a halogen atom and a trifluoromethyl group, a (C1-C4) alkyl group or a (C1-C4) alkoxy group, or an amino group of the general formula NR3R4, wherein R3 and R4 each independently represent a hydrogen atom or a (C1-C4) alkyl group, or form together with the nitrogen atom carrying them a pyrrolidine, piperidine or morpholine ring, or a phenyl group optionally substituted by an atom or group as defined above for symbol X, or a pharmaceutically acceptable salt thereof or a prodrug of said compound or a pharmaceutically acceptable salt thereof.

[0290] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0291] SSR-504734, or a pharmaceutically acceptable salt thereof, or a prodrug of said compound or a pharmaceutically acceptable salt thereof.

[0292] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0293]

[0294] in:

[0295] R1 Yes - (CH2) n -R 1a , wherein n is independently 0-6, and R 1a Selected from:

[0296] (1)C 1-6 Alkyl, which is unsubstituted or substituted by 1 to 6 halogens, hydroxyl groups,

[0297] (2) R 2a 、R 2b and R 2c substituted phenyl,

[0298] (3)C 3-6 Cycloallyl, which is unsubstituted or replaced by C 1-6 Alkyl, 1-6 halogens, hydroxyl or -NR 10 R 11 replace,

[0299] (4)-OC 1-6 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 replace,

[0300] (5)-CO2R 9 ,

[0301] wherein R9 is independently selected from:

[0302] (a) hydrogen,

[0303] (b)-C 1-6 Alkyl, which is unsubstituted or substituted by 1 to 6 fluorine groups,

[0304] (c) benzyl, and

[0305] (d) phenyl,

[0306] (6)-NR 10 R 11 ,

[0307] where R 10 and R 11 Independently selected from:

[0308] (a) hydrogen,

[0309] (b)-C 1-6 Alkyl, which is unsubstituted or substituted by hydroxy, 1-6 fluorine or -NR 12 R 13 Substituted, where R 12 and R 13 are independently selected from hydrogen and -C 1-6 alkyl,

[0310] (c)-C 3-6 Cycloalkyl, which is unsubstituted or substituted by hydroxy, 1-6 fluorine or -NR 12 R 13 replace,

[0311] (d) benzyl,

[0312] (e) phenyl, and

[0313] (7)-CONR 10 R 11 ;

[0314] R 2 Selected from:

[0315] (1) phenyl group, which is R 2a 、R 2b and R 2c replace,

[0316] (2)C 1-8 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy, -NR 10 R 11 , phenyl or heterocyclic ring, wherein the phenyl or heterocyclic ring is replaced by R 2a 、R 2b and R 2c replace,

[0317] (3)C 3-6 Cycloalkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 Replace, and

[0318] (4)-C 1-6 Alkyl-(C 3-6 cycloalkyl), which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 replace;

[0319] R 2a 、R 2b and R 2c Independently selected from:

[0320] (1) Hydrogen,

[0321] (2) halogens,

[0322] (3)-C 1-6 Alkyl, which is unsubstituted or substituted with:

[0323] (a) 1 to 6 halogens,

[0324] (b) phenyl,

[0325] (c)C 3-6 Cycloalkyl, or

[0326] (d)-NR 10 R 11 ,

[0327] (4)-OC 1-6 Alkyl, which is unsubstituted or substituted by 1 to 6 halogens,

[0328] (5) hydroxyl groups,

[0329] (6)-SCF3,

[0330] (7)-SCHF2,

[0331] (8)-SCH3,

[0332] (9)-CO2R 9 ,

[0333] (10)-CN,

[0334] (11)-SO2R 9 ,

[0335] (12)-SO2-NR 10 R 11 ,

[0336] (13)-NR 10 R 11 ,

[0337] (14)-CONR 10 R 11 ,and

[0338] (15)-NO2;

[0339] R 3 Selected from:

[0340] (1)C 1-6 Alkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 replace,

[0341] (2)C 3-6 Cycloalkyl, which is unsubstituted or substituted by 1-6 halogen, hydroxy or -NR 10 R 11 replace,

[0342] R 4 and R 5 Independently selected from:

[0343] (1) hydrogen, and

[0344] (2)C1-6 Alkyl, which is unsubstituted or substituted by halogen or hydroxy, or R 4 and R 5 Together they form C 3-6 cycloalkyl ring;

[0345] A is selected from:

[0346] (1) -O-, and

[0347] (2)-NR 10 -;

[0348] m is 0 or 1, wherein when m is 0, R 2 directly attached to the carbonyl group;

[0349] and pharmaceutically acceptable salts thereof and individual enantiomers and diastereomers thereof, or pharmaceutically acceptable salts thereof or prodrugs of said compounds or pharmaceutically acceptable salts thereof.

[0350] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0351] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0352] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0353]

[0354] in:

[0355] R 1 are independently halogen, C1-C3 alkyl, C3-C6 cycloalkyl, OR 9 or SR 10 Phenyl substituted 1 to 5 times, wherein C1-C3 alkyl and C3-C6 cycloalkyl are optionally replaced by R 7 Replace 1 to 10 times;

[0356] R 2 It is H;

[0357] R 3 and R 4 are each independently H or CH3;

[0358] R 5 Selected from:

[0359] (1) Hydrogen,

[0360] (2) optionally by R 71-C6 alkyl substituted 1 to 11 times,

[0361] (3) geminal dialkyl, and

[0362] (4) Geminal dihalogenated; or

[0363] Two R on the same carbon 5 The substituents, together with the carbon atom to which they are attached, may form a 7 3-, 4- or 5-membered cycloalkyl substituted 1 to 10 times; or

[0364] Two R on adjacent carbons of the ring to which they are attached 5 The substituents together may form an optionally R 7 3-, 4-, 5- or 6-membered cycloalkyl substituted 1 to 10 times;

[0365] R 6 yes

[0366] wherein E, F and G are each independently nitrogen or carbon, and R 6a is C1-C2 alkyl, which is optionally substituted 1 to 5 times by halogen or deuterium;

[0367] R 7 Selected from:

[0368] (1) Hydrogen,

[0369] (2) halogens,

[0370] (3) Deuterium,

[0371] (4) geminal dialkyl,

[0372] (5) Geminal dihalogenated,

[0373] (6)-OR 9 、-NR 11 R 12 、-NR 11 C(O) p R 10 、-S(O) p R 10 、-CN、-NO2、-C(O) p R 10 、-C(O)NR 11 R 12 , or -NR 11 C(S)R 10 ,and

[0374] (7) Oxo or thio;

[0375] R 8 Selected from:

[0376] (1) Hydrogen,

[0377] (2) halogens,

[0378] (3) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, or C4-C7 cycloalkylalkyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each independently and optionally replaced by R 7 Replace 1 to 11 times, or

[0379] (4)-OR 9 、-NR 11 R 12 、-NR 11 C(O) p R 10 、-S(O) p R 10 、-CN、-NO2、-C(O) p R 10 、-C(O)NR 11 R 12 , or -NR 11 C(S)R 10 ;

[0380] R 9 Selected from hydrogen, C1-C4 alkyl, C3-C7 cycloalkyl, C4-C7 cycloalkylalkyl, -C(O)NR 11 R 12 and -C(O) p R 10 , wherein C1-C4 alkyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each optionally replaced by R 7 Replace 1 to 11 times;

[0381] R 10 is selected from hydrogen, C1-C4 alkyl, C3-C7 cycloalkyl, C4-C7 cycloalkylalkyl, aryl and heteroaryl, wherein C1-C4 alkyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each optionally substituted 1 to 11 times with substituents as defined in R7, and aryl or heteroaryl is optionally substituted with R 8 Replace 1 to 10 times;

[0382] R 11 and R 12 are each independently selected from hydrogen, C1-C4 alkyl, C3-C7 cycloalkyl, C4-C7 cycloalkylalkyl, aryl and heteroaryl, wherein C1-C4 alkyl, C3-C7 cycloalkyl and C4-C7 cycloalkylalkyl are each optionally replaced by R 7substituted 1 to 11 times by the substituents defined in 8 Replace 1 to 10 times, or R 11 and R 12 Together with the nitrogen to which they are attached, they form a 7 a saturated or partially saturated monocyclic or fused bicyclic heterocyclic ring substituted 1 to 11 times;

[0383] A is

[0384] X is N;

[0385] Y is N;

[0386] p is 1 or 2; and

[0387] m is 0;

[0388] The conditions are: R 6 cannot be (a) 1H-1,2,3-triazol-4-yl, or (b) 5-methylisoxazol-4-yl;

[0389] or an oxide thereof, a pharmaceutically acceptable salt of the compound or its oxide, or an individual enantiomer or diastereomer thereof.

[0390] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula:

[0391]

[0392]

[0393] A compound or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0394] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a GlyT1 inhibitor having the formula

[0395] or a pharmaceutically acceptable salt thereof or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0396] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0397]

[0398] in:

[0399] R 1represents a phenyl group or a 5- or 6-membered monocyclic heteroaryl group having 1, 2 or 3 heteroatoms independently selected from O, N or S, wherein the phenyl group or heteroaryl group is optionally substituted by one or more R 3 replace;

[0400] R 2 represents an aryl group, a 5- or 6-membered monocyclic heteroaryl group or an 8- to 10-membered bicyclic heteroaryl group, said monocyclic or bicyclic heteroaryl group having 1, 2 or 3 heteroatoms independently selected from O, N or S, wherein said aryl or heteroaryl group is optionally replaced by one or more R 4 replace;

[0401] R 3 Halogen, C 1-4 -alkyl or C 3-6 -cycloalkyl, wherein the C 1-4 -alkyl or the C 3-6 - cycloalkyl is optionally substituted with one or more halogens; and

[0402] R 4 Halogen, -CN, C 1-4 -alkyl, C 3-6 -cycloalkyl, -C 1-3 -alkyl, -C 3-6 -cycloalkyl or -OC 1-6 Alkyl, wherein the C 1-4 -alkyl, C 3-6 -cycloalkyl, -C 1-3 -alkyl, -C 3-6 -cycloalkyl or -OC 1-6 - alkyl is optionally substituted with one or more halogens;

[0403] or a pharmaceutically acceptable salt thereof or a tautomer or stereoisomer of said compound or a pharmaceutically acceptable salt thereof, or a mixture of any of the foregoing.

[0404] In certain embodiments, the compound of Formula IX can be represented by a compound of Formula IX(a): or a pharmaceutically acceptable salt thereof or a tautomer of said compound or a pharmaceutically acceptable salt thereof, or a mixture of any of the foregoing.

[0405] In certain embodiments, the compound of Formula IX can be represented by a compound of Formula IX(b): or a pharmaceutically acceptable salt thereof or a tautomer of said compound or a pharmaceutically acceptable salt thereof, or a mixture of any of the foregoing.

[0406] In certain embodiments, the compound of Formula IX is a compound selected from any one of the following, a stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof:

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420] In some embodiments of the methods and uses disclosed herein, the GlyT1 inhibitor is a compound having the formula

[0421]

[0422] in:

[0423] R 1 Selected from:

[0424] a) a 5- or 6-membered monocyclic heteroaryl group having 1, 2, 3 or 4 heteroatoms independently selected from O, N and S(O)r,

[0425] b) a 5- or 6-membered monocyclic partially saturated heterocycloalkyl group having 1, 2 or 3 heteroatoms independently selected from O, N and S(O)r, and

[0426] c) having 1, 2 or 3 independently selected from O, N and S(O) r a 9- or 10-membered bicyclic heteroaryl group containing a heteroatom

[0427] where r is 0, 1, or 2;

[0428] wherein the groups a), b) and c) are each optionally substituted by one or more substituents independently selected from the group consisting of: 1-4 -alkyl-, C 1-4 -alkyl-O-, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, C 3-6 -cycloalkyl- and C 3-6 -cycloalkyl-O-, and in the case where a substituent is attached to a nitrogen ring atom, said substituent is selected from C 1-4 -alkyl-, C 1-4 -alkyl-CO-, C 3-6 -cycloalkyl- and C 3-6 -cycloalkyl-CO-,

[0429] And wherein said C 1-4 -alkyl-, C 1-4 -alkyl-O-, C 1-4 -alkyl-CO-, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, C 3-6 -cycloalkyl-, C 3-6 -cycloalkyl-CO- or C 3-6 -cycloalkyl-O- substituents may each be substituted with one or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F, and -CN;

[0430] R 2 Selected from hydrogen, C 1-4 -alkyl-, C 1-4 -alkyl-O-, -CN and C 3-6 -cycloalkyl-,

[0431] wherein the C 1-4 -alkyl-, C 1-4 -alkyl-O- and C 3-6 -cycloalkyl- each optionally substituted with 1, 2, 3 or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F and -CN;

[0432] R 3 Selected from C 1-6 -alkyl-O-, C 3-6 -cycloalkyl-O-, morpholino, pyrazolyl and 4 to 7 membered monocyclic heterocycloalkyl-O-, which has 1 oxygen atom as a ring member and optionally 1 or 2 independently selected from O, N and S(O) s heteroatoms, wherein s=0, 1 or 2,

[0433] wherein the C 1-6 -alkyl-O- and the C 3-6-cycloalkyl-O- may be optionally substituted with 1, 2, 3 or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F, -CN, C 1-4 -alkyl-, C 3-6 -cycloalkyl-, C 1-6 -alkyl-O- and C 3-6 -cycloalkyl-O-;

[0434] R 4 It is hydrogen;

[0435] or R 3 and R 4 Together with the ring atoms of the phenyl group to which they are attached, they can form a 4-, 5- or 6-membered monocyclic partially saturated heterocycloalkyl or heteroaryl group, each of which has 1, 2 or 3 atoms independently selected from O, N and S(O) s wherein s=0, 1 or 2, of which in the general formula (I) there must be 1 heteroatom directly attached to R 3 a ring oxygen atom of a ring carbon atom of the phenyl group to which it is attached;

[0436] wherein the heterocycloalkyl group may be optionally substituted by 1, 2, 3 or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F, -CN, C 1-4 -alkyl-, C 3-6 -cycloalkyl-, C 1-6 -alkyl-O-, C 3-6 -cycloalkyl-O-, oxetanyl-O-, tetrahydrofuranyl-O-, and tetrahydropyranyl-O-;

[0437] R 5 It is hydrogen;

[0438] R 6 Selected from hydrogen, C 1-4 -alkyl-SO2-, C 3-6 -cycloalkyl-SO2 and -CN;

[0439] R 7 It is hydrogen;

[0440] or a) R 6 and R 7 or b) R 6 and R 5 One of the pairs together with the ring atoms of the phenyl to which they are attached forms a ring having 1, 2 or 3 atoms independently selected from O, N and S(O) u a 5- or 6-membered partially saturated monocyclic heterocycloalkyl group containing a heteroatom of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 6 a -SO2- member of a ring carbon atom of the phenyl group to which it is attached;

[0441] wherein the heterocycloalkyl group may be optionally substituted by 1, 2, 3 or more substituents independently selected from the group consisting of fluorine, -CF3, -CHF2, -CH2F, -CN, C 1-4 -alkyl-, C 1-6 -alkyl-O- and C 3-6 -cycloalkyl-O- or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0442] In certain embodiments, the compound of Formula X is a compound selected from any one of the following, a stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof:

[0443]

[0444]

[0445]

[0446]

[0447]

[0448] For example, the compound of formula X may be a diastereomeric mixture or a single diastereoisomer of any of the following, or a pharmaceutically acceptable salt thereof:

[0449]

[0450]

[0451]

[0452]

[0453]

[0454]

[0455]

[0456] In certain methods and uses disclosed herein, the subject is a subject in need thereof.

[0457] In some embodiments of the uses and methods disclosed herein, a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a pharmaceutically acceptable salt thereof; or a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a prodrug of a pharmaceutically acceptable salt thereof, is administered in a therapeutically effective amount.

[0458] In some embodiments, the compound or its pharmaceutically acceptable salt, solvate or prodrug is selected from the compounds described herein. Any compound provided herein can be prepared as a pharmaceutically acceptable salt, solvate or prodrug and / or prepared as part of a pharmaceutical composition as described in the patents or patent application publications cited herein.

[0459] Although the compounds described herein may be shown as having a specific stereochemistry around certain atoms, such as cis or trans, the compounds may also be prepared in the opposite orientation or as racemic mixtures. Such isomers or racemic mixtures are included in the present disclosure. In addition, although the compounds are collectively shown in a table, any compound or pharmaceutically acceptable salt, solvate or prodrug thereof may be selected from the table and used in the embodiments provided herein.

[0460] The compounds described herein can be prepared according to the methods described in the patents or patent application publications cited herein.

[0461] The compounds can be used to inhibit the GlyT1 transporter. Thus, in some embodiments, the compounds can be referred to as GlyT1 transporter inhibiting compounds or GlyT1 inhibitors.

[0462] The compounds described herein can be administered in any conventional manner by any route that makes them active. Administration can be systemic, topical or oral. For example, administration can be, but is not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, buccal, sublingual or ocular, or intravaginal, by inhalation, by reservoir injection, or by implant. The mode of administration can depend on the condition or disease to be targeted or treated. The choice of a specific route of administration can be selected or adjusted by the clinician according to methods known to the clinician to obtain the desired clinical response.

[0463] In some embodiments, it may be desirable to administer one or more compounds, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, topically to the area in need of treatment. This can be achieved, for example, but not limited to, by local infusion during surgery, topical application (e.g., in conjunction with a wound dressing after surgery), by injection, by catheter, by suppository, or by implant, wherein the implant is a porous, non-porous, or gel-like material, including membranes, such as silicone rubber membranes or fibers.

[0464] The compounds described herein can be administered alone or in combination (simultaneously or sequentially) with other drugs. For example, the compounds can be administered in combination with other drugs used to treat EPP, XLPP, or CEP, etc. Examples of other drugs or agents are known to those skilled in the art and include, but are not limited to, those described herein.

[0465] Means and methods of administration are known in the art, and the skilled artisan can consult various pharmacology references for guidance (see, e.g., Modern Pharmaceutics, Banker & Rhodes, Marcel Dekker, Inc. (1979); and Goodman & Gilman's The Pharmaceutical Basis of Therapeutics, 6th ed., MacMillan Publishing Co., New York (1980)).

[0466] The amount of compound to be administered is a therapeutically effective amount. The dosage to be administered will depend on the characteristics of the experimenter being treated, such as the specific animal being treated, age, body weight, health status, the type (if any) and the treatment frequency of the treatment being treated, and can be easily determined by those skilled in the art (for example, clinicians). The standard dose of protamine can be used and adjusted (that is, increase or decrease) according to the above factors. The selection of specific dosage regimen can be selected or regulated or titrated by the clinician according to methods known to the clinician, to obtain the clinical response of expectation.

[0467] The amount of the compound described herein for effectively treating and / or preventing a particular disease, illness or disorder will depend on the nature and degree of the disease, illness or disorder, and can be determined by standard clinical techniques. In addition, in vitro or in vivo determinations can optionally be used to help identify the optimal dose range. The exact dosage to be used in the composition will also depend on the severity of the route of administration and the disorder, and should be determined according to the judgment of the practitioner and the situation of each patient. However, the suitable dosage range for oral administration is typically about 0.001 milligram to about 200 milligrams / kg body weight, about 0.01 milligram to about 100 milligrams / kg body weight, about 0.01 milligram to about 70 milligrams / kg body weight, about 0.1 milligram to about 50 milligrams / kg body weight, 0.5 milligram to about 20 milligrams / kg body weight or about 1 milligram to about 10 milligrams / kg body weight. In some embodiments, oral dose is about 5 milligrams / kg body weight.

[0468] In some embodiments, the suitable dosage range for intravenous (iv) use is about 0.01mg to about 500mg / kg body weight, about 0.1mg to about 100mg / kg body weight, about 1mg to about 50mg / kg body weight or about 10mg to about 35mg / kg body weight. Suitable dosage ranges for other modes of administration can be based on above-mentioned dosage calculations well known to those skilled in the art. For example, intranasal, through mucosal, intradermal, intramuscular, intraperitoneal, subcutaneous, epidural, sublingual, intracerebral, intravaginal, transdermal administration or the recommended dose used by inhalation is about 0.001mg to about 200mg / kg body weight, about 0.01mg to about 100mg / kg body weight, about 0.1mg to about 50mg / kg body weight or about 1mg to about 20mg / kg body weight. Effective dose can be extrapolated from the dose-response curve derived from in vitro or animal model test system. Such animal model and system are well known in the art.

[0469] The compounds described herein can be formulated for parenteral administration by injection, such as by push injection or continuous infusion. In some embodiments, the compound can be administered by subcutaneous continuous infusion over a period of about 15 minutes to about 24 hours. Injectable formulations can be presented in unit dosage form, such as in ampoules or multi-dose containers, and are optionally supplemented with preservatives. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle and can contain formulations such as suspending agents, stabilizers, and / or dispersants. In some embodiments, the injectable is a short-acting, depot, or implant and pellet form for subcutaneous or intramuscular injection. In some embodiments, the parenteral dosage form is in the form of a solution, suspension, emulsion, or dry powder.

[0470] For oral administration, the compounds described herein can be formulated by combining the compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds to be formulated into tablets, pills, dragees, capsules, emulsions, liquids, gels, syrups, cachets, pellets, powders, granules, slurries, lozenges, aqueous or oily suspensions, and the like for oral ingestion by the patient to be treated. Pharmaceutical preparations for oral use can be obtained, for example, by adding solid excipients, optionally grinding the resulting mixture, and, if desired, processing the granular mixture after adding suitable adjuvants to obtain tablets or dragee cores. Suitable excipients include, but are not limited to, fillers such as sugars, including but not limited to lactose, sucrose, mannitol, and sorbitol; cellulose preparations such as, but not limited to, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as, but not limited to, the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0471] Orally administered compositions may contain one or more optional agents, such as sweeteners, such as fructose, aspartame, or saccharin; flavorings, such as peppermint, oil of wintergreen, or cherry; coloring agents; and preservatives, to provide pharmaceutically palatable formulations. In addition, in the case of tablet or pill form, the composition may be coated to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect over an extended period of time. Selectively permeable membranes surrounding osmotically active driving compounds are also suitable for orally administered compounds. Oral compositions may contain standard vehicles, such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Such vehicles are preferably pharmaceutical grade.

[0472] Dragee cores may be provided with a suitable coating. For this purpose, concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablets or dragee coatings for identification or characterization of different combinations of active compound dosages.

[0473] Pharmaceutical preparations for oral use include, but are not limited to, push-fit capsules made of gelatin and soft, sealed capsules made of gelatin and a plasticizer (such as glycerol or sorbitol). Push-fit capsules may contain the active ingredient mixed with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, a stabilizer may be added.

[0474] For buccal administration, the compositions may take the form of tablets or lozenges, for example, formulated in conventional manner.

[0475] For administration by inhalation, the compounds described herein can be delivered in the form of an aerosol spray from a pressurized pack or 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 can be determined by providing a valve to deliver a metered amount. Capsules and cartridges such as gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0476] The compounds described herein can also be formulated into rectal compositions, such as suppositories or retention enemas, such as containing conventional suppository bases, such as cocoa butter or other glycerides. The compounds described herein can also be formulated into vaginal compositions, such as vaginal creams, suppositories, pessaries, vaginal rings, and intrauterine devices.

[0477] In transdermal administration, the compound can be applied to a patch or can be applied via a transdermal therapeutic system and subsequently provided to the organism. In some embodiments, the compound is present in a cream, solution, powder, fluid emulsion, fluid suspension, semisolid, ointment, paste, gel, jelly, and foam, or in a patch containing any of these.

[0478] Compounds as described herein can also be formulated into depot preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Depot injections can be administered at intervals of about 1 to about 6 months or longer. Therefore, for example, the compound can be formulated with a suitable polymeric material or hydrophobic material (e.g., as an emulsion in an acceptable oil) or an ion exchange resin, or formulated into a slightly soluble derivative (e.g., formulated into a slightly soluble salt).

[0479] In some embodiments, the compound can be delivered in a controlled release system. In one embodiment, a pump can be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng., 1987, 14, 201; Buchwald et al., Surgery, 1980, 88, 507; Saudek et al., N. Engl. J. Med., 1989, 321, 574). In some embodiments, polymeric materials may be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Press., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger et al., J. Macromol. Sci. Rev. Macromol. Chem., 1983, 23, 61; see also Levy et al., Science, 1985, 228, 190; During et al., Ann. Neurol., 1989, 25, 351; Howard et al., J. Neurosurg., 1989, 71, 105). In yet another embodiment, a controlled release system can be placed near the target of the compounds described herein, such as the liver, thereby requiring only a fraction of the systemic dose (see, e.g., Goodson, Medical Applications of Controlled Release, supra, Vol. 2, pp. 115-138 (1984)). Other controlled release systems discussed in the review by Langer, Science, 1990, 249, 1527-1533 can be used.

[0480] It is also known in the art that the compound can be included in such formulations together with pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic carriers, water-soluble carriers, emulsifiers, buffers, wetting agents, moisturizers, solubilizers, preservatives, etc. The pharmaceutical composition may also include a suitable solid or gel phase carrier or excipient. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycol. In some embodiments, the compounds described herein can be used together with medicaments including, but not limited to, topical analgesics (e.g., lidocaine), barrier devices (e.g., GelClair), or irrigants (e.g., Caphosol).

[0481] In some embodiments, the compounds described herein can be delivered in vesicles, particularly liposomes (see Langer, Science, 1990, 249, 1527-1533; Treat et al., Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, supra, pp. 317-327; see generally supra).

[0482] Suitable compositions include, but are not limited to, oral non-absorbable compositions. Suitable compositions also include, but are not limited to, saline, water, cyclodextrin solutions, and buffered solutions of pH 3-9.

[0483] The compounds described herein, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, can be formulated with a variety of excipients including, but not limited to, purified water, propylene glycol, PEG 400, glycerol, DMA, ethanol, benzyl alcohol, citric acid / sodium citrate (pH 3), citric acid / sodium citrate (pH 5), tris(hydroxymethyl)aminomethane HCl (pH 7.0), 0.9% saline, and 1.2% saline, and any combination thereof. In some embodiments, the excipient is selected from propylene glycol, purified water, and glycerol.

[0484] In some embodiments, the formulation can be lyophilized to a solid and reconstituted with, for example, water, before use.

[0485] When administered to mammals (eg, to animals for veterinary use or to humans for clinical use), the compound can be administered in isolated form.

[0486] When applied to humans, the compound can be sterile. When the compound of formula I-VIII is administered intravenously, water is a suitable carrier. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, glycol, water, ethanol, etc. If necessary, the compositions of the present invention can also contain a small amount of wetting agent or emulsifier or pH buffer.

[0487] The compositions described herein can take the form of solutions, suspensions, emulsions, tablets, pills, pellets, capsules, capsules containing liquids, powders, sustained release formulations, suppositories, aerosols, sprays, or any other form suitable for use. Examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, A.R. Gennaro (ed.) Mack Publishing Co.

[0488] In some embodiments, the compound is formulated into a pharmaceutical composition suitable for administration to a human being according to conventional procedures. Typically, the compound is a solution in a sterile isotonic aqueous buffer. If necessary, the composition may also include a solubilizing agent. Compositions for intravenous administration may optionally include a local anesthetic such as lidocaine to alleviate pain at the injection site. Typically, the ingredients are separated or mixed together and provided in unit dosage form, for example, as a dry lyophilized powder or anhydrous concentrate in an airtight sealed container, the airtight sealed container being, for example, an ampoule or sachet indicating the amount of the active agent. When the compound is administered by infusion, it can be dispensed, for example, with an infusion bottle containing sterile pharmaceutical grade water or saline. When the compound is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed before administration.

[0489] The pharmaceutical compositions can be in unit dosage form. In this form, the composition is divided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparation, such as packaged tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms.

[0490] In some embodiments, the composition is in liquid form, wherein the active agent (i.e., one of the surface amphiphilic polymers or oligomers disclosed herein) is present in the form of a solution, suspension, emulsion, or solution / suspension. In some embodiments, the liquid composition is in the form of a gel. In other embodiments, the liquid composition is aqueous. In other embodiments, the composition is in the form of an ointment.

[0491] In some embodiments, the composition is in the form of a solid product. For example, in some embodiments, the ophthalmic composition is a solid product that can be inserted into a suitable location in the eye, such as between the eye and the eyelid or in the conjunctival sac, where it releases the active agent, as described, for example, in U.S. Patent No. 3,863,633; U.S. Patent No. 3,867,519; U.S. Patent No. 3,868,445; U.S. Patent No. 3,960,150; U.S. Patent No. 3,963,025; U.S. Patent No. 4,186,184; U.S. Patent No. 4,303,637; U.S. Patent No. 5,443,505; and U.S. Patent No. 5,869,079. Typically, the product is released from the cornea via tear fluid that soaks the corneal surface, or directly onto the cornea itself, with the solid product typically being in close contact with the cornea. Solid products suitable for implantation into the eye in this manner are typically composed primarily of a polymer and can be bioerodible or non-bioerodible. Bioerodible polymers that can be used to prepare ocular implants carrying one or more compounds include, but are not limited to, aliphatic polyesters such as poly(glycolide), poly(lactide), poly(ε-caprolactone), poly(hydroxybutyrate), and poly(hydroxyvalerate) polymers and copolymers, polyamino acids, polyorthoesters, polyanhydrides, aliphatic polycarbonates, and polyether lactones. Suitable non-bioerodible polymers include silicone elastomers.

[0492] The compositions described herein may contain a preservative. Suitable preservatives include, but are not limited to, mercury-containing substances such as phenylmercuric salts (e.g., phenylmercuric acetate, phenylmercuric borate, and phenylmercuric nitrate) and thimerosal; stabilized chlorine dioxide; quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride; imidazolidinyl urea; parabens such as methylparaben, ethylparaben, propylparaben, and butylparaben, and their salts; phenoxyethanol; chlorophenoxyethanol; phenoxypropanol; chlorobutanol; chlorocresol; phenylethyl alcohol; disodium EDTA; and sorbic acid and its salts.

[0493] Optionally, one or more stabilizers can be included in the composition to enhance chemical stability when needed. Suitable stabilizers include but are not limited to chelating agents or complexing agents, such as calcium complexing agent ethylenediaminetetraacetic acid (EDTA). For example, an amount of EDTA or its salt, for example disodium salt, can be included in the composition to complex excessive calcium ions and prevent the formation of gel during storage. The appropriate amount of EDTA or its salt can be approximately 0.01% to approximately 0.5%. In those embodiments containing the preservative except EDTA, EDTA or its salt, more particularly disodium EDTA, can exist with the amount of approximately 0.025 % by weight to approximately 0.1 % by weight.

[0494] One or more antioxidants may also be included in the composition. Suitable antioxidants include, but are not limited to, ascorbic acid, sodium metabisulfite, sodium bisulfite, acetylcysteine, polyquaternium-1, benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, phenylethyl alcohol, disodium edetate, sorbic acid, or other agents known to those skilled in the art. Such preservatives are typically used at a level of about 0.001% to about 1.0% by weight.

[0495] In some embodiments, the compound is at least partially solubilized by an acceptable solubilizing agent. Certain acceptable nonionic surfactants (e.g., polysorbate 80) can be used as solubilizing agents, as can ophthalmologically acceptable glycols, polyglycols (e.g., polyethylene glycol 400 (PEG-400)) and glycol ethers.

[0496] Suitable solubilizers for solutions and solution / suspension compositions are cyclodextrins. Suitable cyclodextrins may be selected from α-cyclodextrin, β-cyclodextrin,

[0497] γ-cyclodextrin, alkylcyclodextrin (e.g., methyl-β-cyclodextrin, dimethyl-β-cyclodextrin, diethyl-β-cyclodextrin), hydroxyalkylcyclodextrin (e.g., hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin), carboxy-alkylcyclodextrin (e.g., carboxymethyl-β-cyclodextrin), sulfoalkylethercyclodextrin (e.g., sulfobutylether-β-cyclodextrin), etc. The ophthalmic applications of cyclodextrins are reviewed in Rajewski et al., Journal of Pharmaceutical Sciences, 1996, 85, 1155-1159.

[0498] In some embodiments, the composition optionally contains a suspending agent. For example, in those embodiments where the composition is an aqueous suspension or solution / suspension, the composition may contain one or more polymers as suspending agents. Useful polymers include, but are not limited to, water-soluble polymers such as cellulosic polymers, e.g., hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers.

[0499] The composition may include one or more acceptable pH adjusters and / or buffers, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and tris; and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition within an acceptable range.

[0500] One or more acceptable salts, solvates or prodrugs can be included in the composition in an amount required to bring the osmolality of the composition to an acceptable range. Such salts include, but are not limited to, those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions. In some embodiments, salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate. In some embodiments, the salt is sodium chloride.

[0501] Optionally, one or more acceptable surfactants, such as, but not limited to, nonionic surfactants or cosolvents, may be included in the composition to increase the solubility of the composition components or to impart physical stability, or for other purposes. Suitable nonionic surfactants include, but are not limited to, polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkylphenyl ethers, such as octoxynol 10, octoxynol 40; polysorbates 20, 60, and 80; polyoxyethylene / polyoxypropylene surfactants (e.g., F-68, F84 and P-103); cyclodextrin; or other agents known to those skilled in the art. Typically, these co-solvents or surfactants are used in an amount of about 0.01% to about 2% by weight in the composition.

[0502] In some embodiments, a pharmaceutical package or kit is provided, comprising one or more containers filled with one or more compounds described herein. Optionally associated with such one or more containers may be a notice in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, the notice reflecting approval by the manufacturing, use or sales agency for human administration to treat conditions, diseases or disorders described herein. In some embodiments, the kit contains more than one compound described herein. In some embodiments, the kit comprises a compound described herein in a single injectable dosage form, such as a single dose in an injectable device (such as a syringe with a needle).

[0503] In some embodiments, the method comprises administering to a subject one or more compounds described herein, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, or pharmaceutical compositions thereof. In some embodiments, the subject is a subject in need of such treatment. As described herein, in some embodiments, the subject is a mammal, such as, but not limited to, a human.

[0504] In some embodiments, one or more of the above compounds, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, or pharmaceutical compositions comprising one or more of the above compounds, are also provided for use in the preparation of a medicament for treating and / or preventing EPP, XLPP, or CEP, or a syndrome related thereto, in a subject, including but not limited to the conditions described herein, such as those described herein. In some embodiments, the subject is a subject in need thereof.

[0505] This embodiment also provides the use of one or more of the above compounds or their pharmaceutically acceptable salts, solvates or prodrugs, or pharmaceutical compositions comprising one or more of the above compounds in inhibiting the GlyT1 transporter (such as the presence on the cell surface). In some embodiments, the compound, its pharmaceutically acceptable salt or pharmaceutical composition thereof inhibits the internalization, transport and / or degradation of the GlyT1 transporter.

[0506] As used herein, "inhibit" may refer to the inhibition of a specific activity. The activity of the GlyT1 transporter can be measured by any method known in the art, including but not limited to the methods described herein.

[0507] The compounds described herein are inhibitors of the GlyT1 transporter.The ability of a compound to inhibit GlyT1 transporter activity can be measured using any assay known in the art.

[0508] Generally, assays for testing compounds that inhibit GlyT1 transporter activity include determining any parameter that is indirectly or directly affected by the GlyT1 transporter, such as a functional, physical or chemical effect.

[0509] The sample that comprises the GlyT1 transporter processed with potential inhibitor or mensuration is compared with the control sample that does not contain inhibitor to check the degree of inhibition.The control sample (not processed with inhibitor) is assigned to the relative GlyT1 transporter activity value of 100%.When the GlyT1 transporter activity value relative to the control is about 80%, 50% or 25%, the inhibition of the GlyT1 transporter is realized.

[0510] The combination of ligand and GlyT1 transporter can be tested in various forms. The combination can be carried out in solution, in a bilayer membrane, attached to a solid phase, in a lipid monolayer or in a vesicle. For example, in a determination, the combination of a natural ligand and its transporter is measured in the presence of a candidate modulator (such as a compound as described herein). Alternatively, the combination of a candidate modulator can be measured in the presence of a natural ligand. Typically, competitive assays are used that measure the ability of a compound to compete with a natural ligand for binding to a transporter. Combination can be tested by measuring, for example, the change in spectral characteristics (for example, fluorescence, absorbance, refractive index), the change in fluid dynamics (for example, shape) or the change in chromatogram or solubility characteristics.

[0511] After translocator is expressed in the cell, the cell can be grown in a suitable culture medium in a suitable cell plate. The cell can be, for example, plated in a 384-well plate with 5000-10000 cells / well. In some embodiments, the cell is plated with about 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000 or 10000 cells / well. The plate can have any number of holes, and the number of cells can be changed accordingly.

[0512] Any pharmaceutical agent having utility in the applications described herein can be used for co-treatment, co-administration or co-formulation with the compositions described above.Thus, the compounds described herein can be administered to a subject before, simultaneously with or after such therapeutic agents are administered.

[0513] Additional agents can be administered in co-therapy (including co-formulation) with one or more of the compounds described herein.

[0514] In some embodiments, the response of the disease or disorder to treatment is monitored, and the treatment regimen is adjusted, if necessary, based on such monitoring.

[0515] The frequency of administration is typically such that the dosing interval, e.g., the time period between one dose and the next during waking hours, is about 1 to about 24 hours, about 2 to about 12 hours, about 3 to about 8 hours, or about 4 to about 6 hours. In some embodiments, the dose is administered 1, 2, 3, or 4 times per day. It will be understood by those skilled in the art that the appropriate dosing interval depends, to some extent, on the ability of the selected composition to maintain a concentration of one or more compounds in the subject and / or target tissue (e.g., above the EC 50 Ideally, concentrations above the EC should be maintained for at least 100% of the dosing interval. 50 When this is not possible, it is desirable that concentrations remain above the EC for at least about 60% of the dosing interval. 50, or should remain above the EC for at least about 40% of the dosing interval 50 .

[0516] How to use

[0517] The present application provides a method for preventing or treating a disorder associated with PPIX accumulation in a subject, the method comprising administering to the subject one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or prodrugs of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. In certain embodiments, the glycine transporter inhibitor is a GlyT1 inhibitor, such as a GlyT1 inhibitor disclosed herein. For example, the present application provides a method for preventing or treating a disorder associated with PPIX accumulation in a subject, the method comprising administering to the subject bitopertin, or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or a pharmaceutically acceptable salt thereof.

[0518] In part, the present disclosure relates to methods for treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP), or congenital erythropoietic porphyria (CEP) in a subject, the methods comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In certain embodiments, the present disclosure provides methods for preventing, treating, or reducing the rate of progression and / or severity of one or more complications of EPP, XLPP, or CEP in a subject, the methods comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof. These methods are particularly directed to therapeutic and prophylactic treatment of animals, and more particularly humans. The terms "subject," "individual," or "patient" are interchangeable throughout the specification and refer to humans or non-human animals. These terms include mammals such as humans, non-human primates, laboratory animals, livestock (including cattle, pigs, camels, etc.), companion animals (e.g., dogs, cats, other domestic animals, etc.), and rodents (e.g., mice and rats). In certain embodiments, the patient, subject, or individual is a human.

[0519] The present application provides a method for preventing or treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP) or congenital erythropoietic porphyria (CEP) or its related syndrome (e.g., EPP-related syndrome, XLPP-related syndrome or CEP-related syndrome) in a subject, the method comprising administering to the subject one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or prodrugs of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. The application also provides a method for preventing or treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or prodrugs of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. For example, the present application provides a method for treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or prodrugs of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 and / or GlyT2 inhibitors. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 inhibitors, such as one or more GlyT1 inhibitors disclosed herein. For example, the present application provides a method for preventing or treating EPP, XLPP or CEP in a subject, the method comprising administering bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or a pharmaceutically acceptable salt thereof to the subject.

[0520] The present application also provides a method for preventing or treating EPP, XLPP or CEP or its related syndrome (e.g., EPP-related syndrome, XLPP-related syndrome or CEP-related syndrome) in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or a prodrug of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. The present application also provides a method for preventing or treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or a prodrug of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. For example, the present application provides a method for treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or a prodrug of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 and / or GlyT2 inhibitors. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 inhibitors, such as one or more GlyT1 inhibitors disclosed herein. In certain of the above embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. For example, the present application provides a method for preventing or treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0521] Erythropoietic protoporphyria (EPP) and X-linked protoporphyria (XLPP) are erythropoietic cutaneous porphyrias characterized by acute non-blistering photosensitivity, intolerance to sunlight, and a significantly reduced quality of life. EPP is caused by a partial deficiency of the enzyme ferrochelatase (FECH), which catalyzes the final step in the heme biosynthesis pathway. FECH deficiency increases the level of metal-free erythrocyte PPIX (also referred to herein as "free protoporphyrin IX" and "PPIX"). XLPP is typically caused by C-terminal deletions in the ALAS2 gene that result in gain-of-function mutations. These gain-of-function mutations increase the enzymatic activity of ALAS2 and lead to the accumulation of both metal-free PPIX and zinc-bound PPIX. Both EPP and XLPP result in the accumulation of PPIX in erythrocytes and other tissues or biological fluids (e.g., skin, liver, bile, or feces). PPIX is lipophilic and eliminated through bile, and is hepatotoxic at high concentrations.

[0522] Patients with EPP or XLPP typically develop photosensitivity in early childhood. Patients often experience symptoms of burning, itching, painful erythema, and edema in sun-exposed areas. Skin symptoms are sometimes associated with abnormal liver enzyme activity, hepatobiliary damage such as jaundice and cirrhosis, iron deficiency, and associated microcytic anemia.

[0523] The diagnosis of EPP and XLPP can be determined by measuring the levels of total red blood cells, free protoporphyrin IX, and zinc-protoporphyrin IX in hemolyzed, anticoagulated whole blood. The diagnosis of EPP and / or XLPP can be based on an increase in the level of free protoporphyrin IX in the blood. Patients with XLPP have a significantly higher ratio of zinc-protoporphyrin IX to free protoporphyrin IX (e.g., >25%) compared to patients with EPP (e.g., ≤15%).

[0524] The diagnosis of EPP can also be determined by measuring the level of ferrochelatase activity in a subject. Ferrochelatase is a mitochondrial enzyme that catalyzes the insertion of ferrous iron into PPIX to form heme. Ferrochelatase also catalyzes the insertion of zinc to form zinc protoporphyrin IX (ZPPIX) from any PPIX remaining after heme synthesis is complete. In EPP, free PPIX accumulates in bone marrow reticulocytes because the formation of both heme and ZPPIX is impaired. In some embodiments, the present disclosure relates to methods of treating a subject whose ferrochelatase activity level is reduced to between 10% and 35% of the ferrochelatase activity level observed in a normal subject. In some embodiments, the present disclosure relates to methods of treating a subject whose ferrochelatase activity level is reduced to less than 50% of the ferrochelatase activity level observed in a normal subject.

[0525] XLPP has a similar phenotype to EPP and can be distinguished based on genetic analysis of ALAS2 or by determining the level of ALAS2 enzymatic activity. In some embodiments, the present disclosure relates to methods of treating a subject with a gain-of-function mutation in ALAS2. In some embodiments, the subject has increased ALAS2 enzymatic activity. Because ferrochelatase is not deficient in XLPP, some of the excess PPIX measured in red blood cells is ZPPIX, and a lower percentage (e.g., 50%-85%) is metal-free. In some embodiments, zinc-protoporphyrin IX levels are increased in the subject's red blood cells. In some embodiments, the methods reduce zinc-protoporphyrin IX levels in the subject's red blood cells. In some embodiments, the methods reduce zinc protoporphyrin IX levels in the subject's red blood cells by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0526] In certain aspects, the present disclosure relates to a method of treating erythropoietic protoporphyria (EPP) and / or X-linked protoporphyria (XLPP) in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or a pharmaceutically acceptable salt thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject has increased PPIX levels. In some embodiments, the method relates to a subject having a PPIX level that is at least 10%, 20%, 30%, 40%, or 50% higher than the PPIX level in a healthy subject before administration of the glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the method relates to a subject having a PPIX level that is at least 10% higher than the PPIX level in a healthy subject before administration of the glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the method relates to a subject having a PPIX level that is at least 20% higher than the PPIX level in a healthy subject before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the method relates to a subject having a PPIX level that is at least 30% higher than the PPIX level in a healthy subject before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the method relates to a subject having a PPIX level that is at least 40% higher than the PPIX level in a healthy subject before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the method relates to a subject having a PPIX level that is at least 50% higher than the PPIX level in a healthy subject before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the level of protoporphyrin IX in the feces of the subject increases. In some embodiments, the level of protoporphyrin IX in the skin of the subject increases. In some embodiments, the level of free protoporphyrin IX in the erythrocytes of the subject increases. In some embodiments, the subject has a protoporphyrin IX level in the red blood cells of greater than 31 μmol L-1. In some embodiments, the subject has a protoporphyrin IX level in the red blood cells of 31 μmol L-1 to 53 μmol L-1. In some embodiments, the subject has a protoporphyrin IX level in the red blood cells of greater than 53 μmol L-1.

[0527] The present application also provides a method for inhibiting PPIX synthesis in vivo, the method comprising administering to a subject a glycine transporter inhibitor or a pharmaceutically acceptable salt thereof or a prodrug of one or more glycine transporter inhibitors or a pharmaceutically acceptable salt thereof. In certain aspects, the present disclosure relates to a method for inhibiting PPIX synthesis in vivo, the method comprising administering to a subject a glycine transporter inhibitor (e.g., a GlyT1 inhibitor) or a pharmaceutically acceptable salt thereof or a prodrug of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor) or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure relates to a method for inhibiting PPIX synthesis in vivo by at least 10% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or at least 100%). In some embodiments, the present disclosure relates to a method for inhibiting PPIX synthesis in vivo by at least 20%. In some embodiments, the present disclosure relates to a method for inhibiting PPIX synthesis in vivo by at least 30%. In some embodiments, the present disclosure relates to a method for inhibiting PPIX synthesis in vivo by at least 40%. In some embodiments, the present disclosure relates to methods for inhibiting PPIX synthesis in vivo by at least 50%. In some embodiments, the present disclosure relates to methods for inhibiting PPIX synthesis in vivo by at least 60%. In some embodiments, the present disclosure relates to methods for inhibiting PPIX synthesis in vivo by at least 70%. In some embodiments, the present disclosure relates to methods for inhibiting PPIX synthesis in vivo by at least 80%. In some embodiments, the present disclosure relates to methods for inhibiting PPIX synthesis in vivo by at least 90%. In some embodiments, the present disclosure relates to methods for inhibiting PPIX synthesis in vivo by at least 100%. The present application also provides methods for reducing the rate of PPIX synthesis in vivo, comprising administering to a subject a glycine transporter inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof. In certain embodiments of the methods and uses disclosed herein, PPIX accumulation is inhibited directly or indirectly. In certain such embodiments, PPIX accumulation is inhibited in a dose-dependent manner. In certain embodiments of the foregoing methods, the glycine transporter inhibitor is a GlyT1 inhibitor, such as a GlyT1 inhibitor disclosed herein. For example, the present application provides a method for inhibiting PPIX synthesis in vivo, reducing the rate of PPIX synthesis in vitro and / or inhibiting PPIX accumulation in vivo, comprising administering bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or a pharmaceutically acceptable salt thereof to a subject.

[0528] In some embodiments, the method relates to a method for reducing the level of free protoporphyrin IX in a subject. In some embodiments, the method relates to a method for reducing the level of free protoporphyrin IX in the red blood cells of a subject. In some embodiments, the method reduces the level of protoporphyrin IX in the red blood cells of a subject to a level below 53 μmol L-1. In some embodiments, the method reduces the level of protoporphyrin IX in the red blood cells of a subject to a level below 31 μmol L-1. In some embodiments, the method reduces the level of protoporphyrin IX in the red blood cells of a subject to a level below 15 μmol L-1. In some embodiments, the method relates to reducing the level of protoporphyrin IX in the feces of a subject. In some embodiments, the method reduces the level of protoporphyrin IX in the skin of a subject. In some embodiments, the method involves a method for reducing the free protoporphyrin IX level of a subject by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%). In some embodiments, the method involves a method for reducing the free protoporphyrin IX level of a subject by at least 15%. In some embodiments, the method involves a method for reducing the free protoporphyrin IX level of a subject by at least 20%. In some embodiments, the method involves a method for reducing the free protoporphyrin IX level of a subject by at least 25%. In some embodiments, the method involves a method for reducing the free protoporphyrin IX level of a subject by at least 30%. In some embodiments, the method involves a method for reducing the free protoporphyrin IX level of a subject by at least 35%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 40%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 45%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 50%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 55%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 60%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 65%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 70%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 75%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 80%.In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 85%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 90%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 95%. In some embodiments, the method involves a method of reducing the free protoporphyrin IX level in a subject by at least 100%.

[0529] In certain aspects, the present disclosure relates to a method of treating X-linked protoporphyria (XLPP) in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or a pharmaceutically acceptable salt thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject has increased zinc-protoporphyrin IX (ZPPIX) levels. In some embodiments, the method relates to a subject having a ZPPIX level that is at least 10%, 20%, 30%, 40%, or 50% higher than the ZPPIX level in a healthy subject prior to administration of the glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the method relates to a subject having a ZPPIX level that is at least 10% higher than the ZPPIX level in a healthy subject prior to administration of the glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the methods relate to subjects having a ZPPIX level that is at least 20% higher than the ZPPIX level in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods relate to subjects having a ZPPIX level that is at least 30% higher than the ZPPIX level in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods relate to subjects having a ZPPIX level that is at least 40% higher than the ZPPIX level in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods relate to subjects having a ZPPIX level that is at least 50% higher than the ZPPIX level in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the ZPPIX level in the subject's red blood cells is increased.

[0530] In certain aspects, the present disclosure relates to a method of treating X-linked protoporphyria (XLPP) in a subject, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject has an increased ratio of zinc-protoporphyrin IX (ZPPIX) relative to free protoporphyrin IX (ZPPIX / PPIX ratio) compared to a subject with EPP. In some embodiments, the method relates to a subject having a ZPPIX / PPIX ratio of at least 15% (e.g., 15%, 20%, 25%, 30%, 35%, 40%, or 45%). In some embodiments, the method relates to a subject having a ZPPIX / PPIX ratio of at least 20%. In some embodiments, the method relates to a subject having a ZPPIX / PPIX ratio of at least 25%. In some embodiments, the methods involve subjects having a ZPPIX / PPIX ratio of at least 30%. In some embodiments, the methods involve subjects having a ZPPIX / PPIX ratio of at least 35%. In some embodiments, the methods involve subjects having a ZPPIX / PPIX ratio of at least 40%. In some embodiments, the methods involve subjects having a ZPPIX / PPIX ratio of at least 45%.

[0531] In certain aspects, the present disclosure relates to methods of inhibiting zinc protoporphyrin IX (ZPPIX) synthesis in vivo, comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 10% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or at least 100%). In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 20%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 30%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 40%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 50%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 60%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 70%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 80%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 90%. In some embodiments, the present disclosure relates to methods of inhibiting ZPPIX synthesis in vivo by at least 100%.

[0532] In certain aspects, the present disclosure relates to a method for treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP), or congenital erythropoietic porphyria (CEP) in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or a pharmaceutically acceptable salt thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject has increased 5-aminolevulinic acid (5-ALA) levels. In some embodiments, the method relates to a subject having a 5-ALA level that is at least 10%, 20%, 30%, 40%, or 50% higher than the 5-ALA level in a healthy subject before administering the glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the method relates to a subject having a 5-ALA level that is at least 10% higher than the 5-ALA level in a healthy subject before administering the glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the methods relate to subjects with 5-ALA levels that are at least 20% higher than 5-ALA levels in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods relate to subjects with 5-ALA levels that are at least 30% higher than 5-ALA levels in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods relate to subjects with 5-ALA levels that are at least 40% higher than 5-ALA levels in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods relate to subjects with 5-ALA levels that are at least 50% higher than 5-ALA levels in healthy subjects before administering a glycine transporter inhibitor (e.g., a GlyT1 inhibitor).

[0533] In certain aspects, the present disclosure relates to methods of inhibiting 5-aminolevulinic acid (5-ALA) synthesis in vivo, comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 10% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or at least 100%). In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 20%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 30%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 40%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 50%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 60%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 70%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 80%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 90%. In some embodiments, the present disclosure relates to methods of inhibiting 5-ALA synthesis in vivo by at least 100%.

[0534] The application also provides one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or the one or more glycine transporter inhibitors or prodrugs of their pharmaceutically acceptable salts in the preparation of a formulation for treating EPP, XLPP, CEP or its related syndromes (e.g., EPP-related syndrome, XLPP-related syndrome or CEP-related syndrome) in a subject. In some embodiments, the application provides one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or the one or more glycine transporter inhibitors or prodrugs of their pharmaceutically acceptable salts in the preparation of a formulation for treating EPP, XLPP or CEP in a subject. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 and / or GlyT2 inhibitors. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 inhibitors, such as one or more GlyT1 inhibitors disclosed herein. In certain such embodiments, the GlyT1 inhibitor is bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or its pharmaceutically acceptable salt. In certain of the foregoing embodiments, the formulation is administered in a therapeutically effective amount.

[0535] The present application provides the use of one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or prodrugs of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof in the preparation of a pharmaceutical composition for treating EPP, XLPP, or CEP or its related syndromes (e.g., EPP-related syndrome, XLPP-related syndrome or CEP-related syndrome) in a subject. In some embodiments, the present application provides the use of one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof or prodrugs of the one or more glycine transporter inhibitors or pharmaceutically acceptable salts thereof in the preparation of a pharmaceutical composition for treating EPP, XLPP or CEP in a subject. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 and / or GlyT2 inhibitors. In some embodiments, the one or more glycine transporter inhibitors are one or more GlyT1 inhibitors, such as one or more GlyT1 inhibitors disclosed herein. In certain such embodiments, the GlyT1 inhibitor is bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or a pharmaceutically acceptable salt thereof. In certain of the above embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0536] Congenital erythropoietic porphyria (CEP) is an erythropoietic cutaneous porphyria characterized by blistering cutaneous photosensitivity. Severe cases of CEP can present in utero with hydrops fetalis or shortly after birth with severe blistering photosensitivity, red urine, splenomegaly, hemolysis, and transfusion dependency. Milder cases and later-onset forms typically present with red urine, severe blistering, and hemolytic anemia.

[0537] Individuals with CEP are typically homozygous or compound heterozygous for UROS mutations. Some cases of CEP are due to mutations in the gene encoding the transcriptional regulator GATA1. These mutations result in reduced activity of uroporphyrinogen III synthase (UROIII-S), the fourth enzyme in the heme biosynthesis pathway. Reduced UROIII-S activity leads to the accumulation of hydroxymethylbilin, which spontaneously forms uroporphyrinogen I, which is further metabolized to coproporphyrinogen I. Uroporphyrinogen I and coproporphyrinogen I accumulate in tissues.

[0538] The diagnosis of CEP can be determined by analyzing the enzymatic activity of uroporphyrinogen III synthase (UROIII-S), by assessing mutations in the UROS gene, by assessing the function of the GATA-1 erythroid-specific transcription factor, by assessing mutations in GATA1, and by determining the uroporphyrin I and coproporphyrin I levels of the subject. In some embodiments, the subject has a mutation in UROS. In some embodiments, the subject has a gene defect in the GATA-1 erythroid-specific transcription factor. In some embodiments, the method relates to a method for treating a subject, wherein the subject has reduced uroporphyrinogen III synthase activity. In some embodiments, increased uroporphyrin I and / or coproporphyrin I levels are measured in the urine or erythrocytes of the subject. In some embodiments, increased coproporphyrin I levels are measured in the feces of the subject.

[0539] In certain aspects, the present disclosure relates to methods of treating congenital erythropoietic porphyria (CEP) in a subject, the methods comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject has increased levels of uroporphyrin I and / or coproporphyrin I. In some embodiments, the subject has increased levels of uroporphyrin I and / or coproporphyrin I. In some embodiments, the methods relate to subjects who have uroporphyrin I levels that are at least 10%, 20%, 30%, 40%, or 50% higher than the levels of uroporphyrin I in healthy subjects prior to administration of a glycine transporter inhibitor (e.g., GlyT1 inhibitor). In some embodiments, the methods involve subjects who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a uroporphyrin I level that was at least 10% higher than the uroporphyrin I level of a healthy subject. In some embodiments, the methods involve subjects who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a uroporphyrin I level that was at least 20% higher than the uroporphyrin I level of a healthy subject. In some embodiments, the methods involve subjects who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a uroporphyrin I level that was at least 30% higher than the uroporphyrin I level of a healthy subject. In some embodiments, the methods involve subjects who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a uroporphyrin I level that was at least 40% higher than the uroporphyrin I level of a healthy subject. In some embodiments, the methods involve a subject having a uroporphyrin I level that is at least 50% higher than a healthy subject's uroporphyrin I level prior to administration of a glycine transporter inhibitor (eg, a GlyT1 inhibitor).

[0540] In some embodiments, the present disclosure relates to methods of treating a subject who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a coproporphyrin I level that was at least 10%, 20%, 30%, 40%, or 50% higher than the coproporphyrin I level of a healthy subject. In some embodiments, the methods relate to a subject who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a coproporphyrin I level that was at least 10% higher than the coproporphyrin I level of a healthy subject. In some embodiments, the methods relate to a subject who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a coproporphyrin I level that was at least 20% higher than the coproporphyrin I level of a healthy subject. In some embodiments, the methods relate to a subject who, prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor), had a coproporphyrin I level that was at least 30% higher than the coproporphyrin I level of a healthy subject. In some embodiments, the methods involve subjects having coproporphyrin I levels that are at least 40% higher than the coproporphyrin I levels of healthy subjects prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor). In some embodiments, the methods involve subjects having coproporphyrin I levels that are at least 50% higher than the coproporphyrin I levels of healthy subjects prior to administration of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor).

[0541] In certain aspects, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I and / or coproporphyrin I in vivo, comprising administering to a subject a glycine transporter inhibitor (e.g., a GlyT1 inhibitor) or a pharmaceutically acceptable salt thereof, or a prodrug of a glycine transporter inhibitor (e.g., a GlyT1 inhibitor) or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 10% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or at least 100%). In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 20%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 30%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 40%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 50%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 60%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 70%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 80%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 90%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of uroporphyrin I in vivo by at least 100%.

[0542] In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 10% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or at least 100%). In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 20%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 30%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 40%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 50%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 60%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 70%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 80%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 90%. In some embodiments, the present disclosure relates to methods of inhibiting the synthesis of coproporphyrin I in vivo by at least 100%.

[0543] Porphyrins (e.g., PPIX, ZPPIX, uroporphyrin I and coproporphyrin I) can be found in various biological samples including skin, urine, feces, blood plasma and red blood cells. In some embodiments, porphyrins can be extracted from biological samples into a solution for fluorescence analysis. Porphyrins can be detected in these biological samples by direct inspection using long wavelength ultraviolet light (e.g., 400-420nm light). Porphyrins have a maximum absorption wavelength near 400-420nm, with their highest absorption peak occurring at 415nm. The emission maximum of porphyrins is typically about 600nm and varies slightly based on the type of porphyrin and the solvent used for analysis. In some embodiments, the diagnosis of EPP, XLPP and CEP can be performed using fluorescence analysis. In some embodiments, skin porphyrin levels (e.g., PPIX levels) can be measured by calculating the difference before and after complete photobleaching of PPIX using controlled illumination. See, e.g., Heerfordt IM. Br J Dermatol. 2016; 175(6): 1284-1289.

[0544] In some embodiments, when irradiated with blue light (e.g., 400-420nm light), the plasma porphyrin of the experimenter fluoresces at a peak value of 634nm. In some embodiments, when irradiated with blue light (e.g., 400-420nm light), the plasma porphyrin of the experimenter fluoresces at a peak value of 626nm and 634nm. In some embodiments, when irradiated with blue light (e.g., 400-420nm light), the skin porphyrin of the experimenter fluoresces at a peak value of 632nm. In some embodiments, when irradiated with blue light (e.g., 400-420nm light), the skin porphyrin of the experimenter fluoresces at a peak value of 626nm and 634nm. In some embodiments, in the skin of the experimenter, protoporphyrin IX level is greater than 0.2 fluorescence units (FDU). In some embodiments, in the skin of the experimenter, protoporphyrin IX level is greater than 1.0FDU. In some embodiments, the protoporphyrin IX level in the subject's skin is between 1.0 FDU and 2.5 FDU. In some embodiments, the protoporphyrin IX level in the subject's skin is greater than 2.5 FDU. In some embodiments, the method reduces the protoporphyrin IX level in the subject's skin to less than 0.5 FDU. In some embodiments, the method reduces the protoporphyrin IX level in the subject's skin to less than 1.0 FDU. In some embodiments, the method reduces the protoporphyrin IX level in the subject's skin to less than 1.5 FDU. In some embodiments, the method reduces the protoporphyrin IX level in the subject's skin to less than 2.0 FDU. In some embodiments, the method reduces the protoporphyrin IX level in the subject's skin to less than 2.5 FDU. In some embodiments, the subject has red fluorescent urine. In some embodiments, using plasma porphyrin fluorescence analysis, the subject has a peak between 615 nm and 620 nm.

[0545] In certain aspects, the present disclosure relates to a method for preventing, treating, or reducing the rate of progression and / or severity of one or more complications of EPP, XLPP, or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the one or more complications of EPP, XLPP, or CEP are selected from the group consisting of acute photosensitivity, cutaneous photosensitivity, edema, erythema, anemia, hypochromic anemia, hemolytic anemia, hemolysis, mild hemolysis, severe hemolysis, chronic hemolysis, hypersplenism, palmar keratosis, bullae, lesions, scars, deformities, nail loss, toe loss, cholelithiasis, cholestasis, cell lysis, gallstones, cholestatic liver failure, erythrodontia, hypercellular bone marrow, myelodysplasia, thrombocytopenia, fetal hydrops, and / or intrauterine death. In some embodiments, the present disclosure contemplates methods of treating one or more complications of EPP, XLPP, or CEP (e.g., acute photosensitivity, cutaneous photosensitivity, edema, erythema, anemia, hypochromic anemia, hemolytic anemia, hemolysis, mild hemolysis, severe hemolysis, chronic hemolysis, hypersplenism, palmar keratosis, bullae, lesions, scars, deformities, nail loss, toe loss, cholelithiasis, cholestasis, cell lysis, gallstones, cholestatic liver failure, erythrodontia, high cellularity bone marrow, myelodysplasia, thrombocytopenia, hydrops fetalis, and / or intrauterine death) comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, one or more complications are indirectly ameliorated. In some embodiments, the present disclosure contemplates a method for preventing one or more complications of EPP, XLPP, or CEP, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the present disclosure contemplates a method for reducing the rate of progression of one or more complications of EPP, XLPP, or CEP, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof.In some embodiments, the present disclosure contemplates methods of reducing the severity of one or more complications of EPP, XLPP, or CEP, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof.

[0546] Optionally, the methods disclosed herein for preventing, treating, or reducing the rate of progression and / or severity of one or more complications of EPP, XLPP, or CEP in a subject may further comprise administering to the patient one or more supportive therapies or additional active agents for treating EPP, XLPP, or CEP. For example, the patient may also be administered one or more supportive therapies or active agents selected from the group consisting of: sun avoidance, topical sunscreen, skin protection, UVB phototherapy, afamelanotide, Bortezomib, proteasome inhibitors, chemical chaperones, cholestyramine, activated charcoal, iron supplementation, liver transplantation, bone marrow transplantation, splenectomy, and blood transfusion. In some embodiments, the methods described herein may further comprise administering afamelanotide to the patient.

[0547] Photosensitization of porphyrins in EPP, XLPP, and CEP results in two distinct clinical syndromes: (1) acute photosensitivity to sun exposure with erythema and edema, and (2) a syndrome in which subepidermal bullae appear in sun-exposed areas of the skin. In certain aspects, the present disclosure relates to methods for preventing, treating, or reducing the rate of progression and / or severity of EPP, XLPP, or CEP in a subject, the methods comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the methods increase the subject's pain-free light exposure. In some embodiments, the methods increase the subject's pain-free light exposure by at least 10%, 20%, 30%, 40%, or 50% more than the pain-free light exposure prior to administration of the GlyT1 inhibitor. In some embodiments, the methods reduce the subject's photosensitivity. In some embodiments, the method reduces the subject's photosensitivity by at least 10%, 20%, 30%, 40%, or 50% compared to photosensitivity before administration of the GlyT1 inhibitor. In some embodiments, the subject has a history of phototoxic reactions from EPP. In some embodiments, the subject is an adult, child, infant, or pregnant woman.

[0548] Glycine is one of the key starting substrates for heme and globin synthesis. Therefore, reduced glycine levels due to GlyT1 inhibition may lead to reduced heme synthesis. In certain aspects, the present disclosure relates to a method for treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject's hemoglobin level is reduced by no more than 10% (e.g., 10%, 15%, 20%, 25% and 30%). In some embodiments, the present disclosure relates to a method for treating EPP, XLPP or CEP in a subject, wherein the subject's hemoglobin level is reduced by no more than 15%. In some embodiments, the present disclosure relates to a method for treating EPP, XLPP or CEP in a subject, wherein the subject's hemoglobin level is reduced by no more than 20%. In some embodiments, the present disclosure relates to methods of treating EPP, XLPP, or CEP in a subject, wherein the subject's hemoglobin levels are reduced by no more than 25%. In some embodiments, the present disclosure relates to methods of treating EPP, XLPP, or CEP in a subject, wherein the subject's hemoglobin levels are reduced by no more than 30%.

[0549] In certain aspects, the present disclosure relates to a method for treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the subject's PPIX level is reduced while the patient's hemoglobin level is significantly maintained. In some embodiments, the patient's PPIX level is reduced by at least 50% (e.g., 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%) and the patient's hemoglobin level is reduced by no more than 10% (e.g., 10%, 15%, 20%, 25% and 30%). In some embodiments, the patient's PPIX level is reduced by at least 85% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 80% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 75% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 70% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 65% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 60% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 55% and the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's PPIX level is reduced by at least 50% and the patient's hemoglobin level is reduced by no more than 15%.

[0550] In certain aspects, the present disclosure relates to a method of treating EPP, XLPP, or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof, wherein the dose of the pharmaceutical composition does not result in a significant decrease in hemoglobin levels. In some embodiments, the patient's PPIX level is reduced by at least 50% (e.g., 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%). In some embodiments, the patient's PPIX level is reduced by at least 55%. In some embodiments, the patient's PPIX level is reduced by at least 60%. In some embodiments, the patient's PPIX level is reduced by at least 65%. In some embodiments, the patient's PPIX level is reduced by at least 70%. In some embodiments, the patient's PPIX level is reduced by at least 75%. In some embodiments, the patient's PPIX level is reduced by at least 80%. In some embodiments, the patient's PPIX level is reduced by at least 85%. In some embodiments, the patient's PPIX level is reduced by at least 90%. In some embodiments, the patient's PPIX level is reduced by at least 95%. In some embodiments, the patient's PPIX level is reduced by at least 100%. In some embodiments, the patient's hemoglobin level is reduced by no more than 10% (e.g., 10%, 15%, 20%, 25% and 30%). In some embodiments, the patient's hemoglobin level is reduced by no more than 15%. In some embodiments, the patient's hemoglobin level is reduced by no more than 20%. In some embodiments, the patient's hemoglobin level is reduced by no more than 25%. In some embodiments, the patient's hemoglobin level is reduced by no more than 30%.

[0551] In some embodiments, the accumulation of one or more of the following heme intermediates is inhibited, wherein the one or more heme intermediates are selected from PPIX, ZPPIX, uroporphyrin I, coproporphyrin I and / or 5-ALA. In some embodiments, the present disclosure relates to a method of inhibiting PPIX accumulation, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the present disclosure relates to a method of inhibiting ZPPIX accumulation, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the present disclosure relates to a method of inhibiting the accumulation of uroporphyrin I, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the present disclosure relates to a method of inhibiting the accumulation of coproporphyrin I, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the present disclosure relates to a method of inhibiting the accumulation of 5-ALA, comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or prodrugs of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the accumulation of one or more heme intermediates (e.g., PPIX, ZPPIX, uroporphyrin I, coproporphyrin I, and / or 5-ALA) is inhibited in a dose-dependent manner. See, e.g., Figure 7 .

[0552] When the liver load exceeds the canalicular excretion capacity, the accumulation of protoporphyrin in EPP, XLPP, and CEP may cause liver damage. Accumulation of PPIX in hepatocytes and bile ductules may lead to cell damage, cholestasis, cell lysis, and further retention of protoporphyrin. Excess protoporphyrin can exert a cholestatic effect, leading to changes in the hepatobiliary system, which can range from mild inflammation to fibrosis and cirrhosis (e.g., cholelithiasis, mild liver disease, worsening liver disease, and end-stage liver disease). Between 3% and 5% of patients with EPP or XLPP develop protoporphyria liver disease, a serious liver disease that may progress rapidly and require liver transplantation. Approximately 2% of patients will develop severe liver disease.

[0553] In certain aspects, the present disclosure relates to a method for preventing, treating, or reducing the rate of progression and / or severity of a liver disease associated with EPP, XLPP, or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or pharmaceutically acceptable salts thereof, or a prodrug of the one or more glycine transporter inhibitors (e.g., GlyT1 inhibitors) or salts thereof. In some embodiments, the liver disease associated with EPP, XLPP, or CEP is cholelithiasis. In some embodiments, the liver disease associated with EPP, XLPP, or CEP is mild liver disease. In some embodiments, the liver disease associated with EPP, XLPP, or CEP is exacerbated liver disease. In some embodiments, the liver disease associated with EPP, XLPP, or CEP is end-stage liver disease.

[0554] The liver function of EPP, XLPP and CEP patients can be assessed using various known clinical assays. In some embodiments, liver function tests can be used to determine the levels of various biochemical parameters (e.g., elevated aspartate aminotransferase levels, alkaline phosphatase or gamma-glutamyl transferase levels). In some embodiments, the histopathology of a liver biopsy can be used to assess one or more parameters of a subject (e.g., protoporphyrin deposition, fibrosis, infiltration, portal fibrosis and periportal fibrosis). In some embodiments, ultrastructural studies of biopsy specimens can be used to determine whether there are vacuoles containing crystals in a subject. Along with the deterioration of liver function, urinary coproporphyrin excretion increases. In some embodiments, coproporphyrin excretion in urine can be analyzed to assess the liver function of a subject. In some embodiments, ultrasound or magnetic resonance elastography can be used to measure the liver stiffness of a subject.

[0555] In certain embodiments of the methods and uses disclosed herein, a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a pharmaceutically acceptable salt thereof; or a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a prodrug of a pharmaceutically acceptable salt thereof, exhibits PPIX inhibition with an EC50 of less than 500 nM, less than 400 nM, less than 300 nM, less than 200 nM, or less than 100 nM. In certain embodiments of the present application, a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a pharmaceutically acceptable salt thereof; or a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a prodrug of a pharmaceutically acceptable salt thereof, exhibits PPIX inhibition with an EC50 of less than 100 nM. In certain embodiments of the present application, a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a pharmaceutically acceptable salt thereof; or a glycine transporter inhibitor, such as a GlyT1 inhibitor (e.g., a GlyT1 inhibitor as disclosed herein) or a prodrug of a pharmaceutically acceptable salt thereof exhibits PPIX inhibition with an EC50 of less than 50 nM. In certain such embodiments, the EC50 is measured in a flow cytometry assay. In certain of the foregoing embodiments, the GlyT1 inhibitor is bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of bitopertin or a pharmaceutically acceptable salt thereof.

[0556] In certain embodiments of the methods and uses disclosed herein, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% cell viability is maintained. In certain such embodiments, at least 90% cell viability is maintained.

[0557] This disclosure also provides the following non-limiting embodiments:

[0558] In order to more efficiently understand the embodiments disclosed herein, examples are provided below. It should be understood that these examples are for illustrative purposes only and should not be construed as limiting the embodiments in any way. Unless otherwise stated, throughout these examples, there may be molecular cloning reactions and other standard recombinant DNA techniques described, which are carried out using commercially available reagents according to the methods described in Maniatis et al., Molecular Cloning-A Laboratory Manual, 2nd edition., Cold Spring Harbor Press (1989).

[0559] The following examples illustrate but do not limit the methods and compositions described herein. Other suitable modifications and adaptations of the various conditions and parameters normally encountered in the therapeutic, synthetic, and other embodiments disclosed herein are within the spirit and scope of the embodiments.

[0560] Example

[0561] Example 1: Synthesis of compounds

[0562] Compounds disclosed herein can be prepared according to well-known procedures and by methods known and disclosed in the art. For example, compounds of formula I, such as bitopertin, can be prepared according to the synthesis schemes provided in U.S. Patent Nos. 7,319,099, 9,877,963, and 7,812,161, the contents of which are incorporated herein by reference in their entirety. In addition, compounds of formula II, such as PF-3463275, can be prepared according to the synthesis schemes provided in U.S. Patent No. 8,124,639, the contents of which are incorporated herein by reference in their entirety.

[0563] Example 2: Treatment of patients with erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP) and congenital GlyT1 inhibitors for subjects with erythropoietic porphyria (CEP). (Prophetic Example)

[0564] To support the production of large amounts of hemoglobin, the synthesis of large amounts of heme is a fundamental requirement for developing red blood cells. In this cell lineage, the amount of heme required to meet this demand far exceeds that of any other cell type. Heme synthesis begins by condensing glycine with succinyl-CoA via the enzyme ALAS. This is the rate-limiting step in heme biosynthesis, ensuring that heme intermediates do not accumulate and cause toxicity. Erythroid cells have acquired an erythroid-specific form of ALAS (ALAS2) and the glycine transporter GlyT1 to increase glycine availability and meet this high demand for heme.

[0565] Animal and human studies that eliminate GlyT1 activity by gene deletion (Garcia-Santos et al., 2017) or reduce GlyT1 activity by administering specific GlyT1 inhibitors (Pinard et al., 2018) have shown that heme synthesis in erythroid cells is reduced, resulting in a moderate microglobulin hypochromic anemia as a result of impaired hemoglobin production. These findings suggest that regulation of glycine uptake in erythrocytes can regulate the heme biosynthesis pathway.

[0566] In patients with erythropoietic protoporphyria or congenital erythropoietic porphyria, specific mutations in single genes encoding enzymes of the heme biosynthetic pathway result in altered enzyme activity and accumulation of heme intermediates upstream of the affected enzyme. Accumulation of these metabolites occurs because the mutant enzyme becomes the rate-limiting step in the pathway and its activity is insufficient to completely convert the upstream metabolite to the next step in the pathway. Three disorders are of particular concern:

[0567] 1. EPP is caused by mutations in the ferrochelatase gene that result in decreased enzyme activity and accumulation of the upstream metabolite protoporphyrin IX (PPIX). An acquired form of EPP can rarely be observed in elderly individuals who have developed new clones containing ferrochelatase mutations as a characteristic feature of myelodysplasia.

[0568] 2. XLPP caused by activating mutations in the ALAS2 gene, which results in high levels of PPIX. In this case, because ferrochelatase cannot fully convert heme into heme even at normal levels, the metabolite accumulates downstream of the affected enzyme.

[0569] 3. CEP is caused by mutations in the uroporphyrinogen synthase gene, which leads to decreased enzyme activity and accumulation of the upstream metabolite coproporphyrin I.

[0570] These heme intermediates can escape from erythrocytes and cause toxicity by hemolysis (in CEP) or by active transport out of the cell (in EPP and XLPP). A consistent feature of all three diseases is a severe, painful, blistering skin reaction after sun exposure, which leads to permanent scarring and deformity. This is caused by the local production of active intermediates by the action of sunlight on PPIX or coproporphyrin I, which triggers a severe inflammatory response. PPIX is hydrophobic and is therefore excreted through the bile duct. High bile concentrations may lead to cholelithiasis, cholestasis, and severe liver damage, leading to liver failure. In the case of CEP, the accumulation of coproporphyrins in mature erythrocytes may lead to severe hemolytic anemia.

[0571] These disease manifestations of EPP, XLPP, and CEP are caused by overproduction of intermediate heme metabolites due to genetic abnormalities in the heme biosynthetic pathway. The accumulated metabolites accumulate in the skin and are toxic to red blood cells after sun exposure or due to bile excretion by the liver. GlyT1 controls the availability of one of the initial substrates in the heme biosynthetic pathway and has been shown to downregulate heme production in humans or animals with normal heme pathways as described above. Without being bound by any particular theory, GlyT1 can similarly reduce the production of intermediate metabolites of heme, particularly when those intermediates accumulate due to abnormal enzyme activity. Therefore, treating subjects with EPP, XLPP, or CEP with GlyT1 will reduce the production of toxic metabolites in erythroid cells in such subjects and lead to reduced skin accumulation of these metabolites, reduced hepatobiliary excretion, or, in the case of CEP, reduced hemolysis, all resulting in reduced disease severity. Thus, the disease is treated.

[0572] Example 3: MetGlyT1 inhibitors effectively reduce the expression of erythroleukemia cells containing EPP, XLPP or CEP pathogenic mutations Levels of heme metabolites in the system.

[0573] Erythroleukemia cells are genetically modified to create cell lines containing pathogenic mutations in EPP, XLPP, or CEP. These genetically modified cell lines are treated with GlyT1 inhibitors, and the production of heme metabolites is assessed photometrically, biochemically, or in radiolabeled studies. The level of photohemolysis caused by PPIX is assessed in these cell lines and found to be reduced in the presence of GlyT1 inhibitors.

[0574] Example 4: GlyT1 inhibitors effectively reduce hemoglobin in erythrocytes containing EPP, XLPP or CEP pathogenic mutations Levels of metabolites. (Prophetic Example)

[0575] Erythroid cells are obtained from the bone marrow or peripheral blood of animals with pathogenic mutations in specific genes that cause EPP, XLPP, or CEP. These cell lines are treated with GlyT1 inhibitors, and the production of heme metabolites is assessed photometrically, biochemically, or in radiolabeled studies. The level of photohemolysis caused by PPIX is assessed in these cell lines and found to be reduced in the presence of GlyT1 inhibitors.

[0576] Example 5: GlyT1 inhibitors effectively reduce hemoglobin in red blood cells of patients with EPP, XLPP or CEP pathogenic mutations Levels of erythroid metabolites. (Prophetic Example)

[0577] Erythroid cells (reticulocytes and erythrocytes) were obtained from patients with EPP, XLPP, and CEP (if available). Cells from patients were treated with GlyT1 inhibitors and the production of heme metabolites was assessed photometrically, biochemically, or in radiolabeled studies. The level of photohemolysis caused by PPIX was assessed in these cell lines and found to be reduced in the presence of GlyT1 inhibitors.

[0578] Example 6: GlyT1 inhibitors effectively reduce the severity of EPP or XLPP in animals. (Prophetic Example)

[0579] Animals with EPP and XLPP were treated with one or more GlyT1 inhibitors at varying doses over time and found to have reduced levels of toxic heme intermediates and improved severity of symptoms of these diseases, such as skin reactions, hepatobiliary disease, and / or hemolysis.

[0580] The embodiments and examples provided herein demonstrate that GlyT1 inhibitors can be used to treat EPP, XLPP or CEP. This is a surprising and unexpected result.

[0581] Example 7: EPP cell model

[0582] Knockout guide sequences were designed to target exon 3 of the ferrochelatase gene. The guide sequences tested are shown in Table 1.

[0583] Table 1: Tested guide sequences

[0584]

[0585] K562 cells were cultured in Iscove's modified Dulbecco's medium (IMDM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin / streptomycin (PS). CRISPR Cas9 RNP complexes with guide RNA were electroporated in K562 cells. Genomic DNA was isolated from the pooled cells, amplified by PCR, and sequenced by Sanger sequencing to determine the knockout efficiency. Single cell clones were isolated by fluorescence-assisted cell sorting (FACS). TA cloning and Sanger sequencing were used to confirm single cell clones and genotypes. Five clones (clone IDs: clone 1-7; clone 1-9; clone 1-10, clone 1-32; clone 1-51; and K562 WT) were selected for further characterization by Western blotting ( Figure 1 ) to determine the FECH protein expression level (antibody: FECH antibody rabbit polyclonal, Proteintech, 14466-1-AP) and PPIX level by flow cytometry ( Figure 2 LC / MS / MS confirmed the accumulation of PPIX in clones 1-9 compared to WT K562 cells ( Figure 3 The genotypic characteristics of the five clones are provided in Table 2.

[0586] Table 2: Genotypic characteristics of clones

[0587] <![CDATA[ Clone ID ]]> <![CDATA[ genotype ]]> K562WT WT / WT 1-7 KO / KO 1-9 Ko / missense T81H 1-10 KO / WT 1-32 KO / WT 1-51 KO / WT

[0588] Will be 2x10 5900 μL of K562 clone-9 cells in IMDM medium containing 10% FBS and 1% PS were plated into 24-well plates. After 24 hours of incubation, 100 μL of compound in DMSO / medium at varying concentrations was added. The final DMSO concentration was 0.1%. The compounds were incubated at 37°C for 96 hours. Cell viability and cell counts were measured using the Vi-CELL XR Complete System. Finally, the effects of the compounds on PPIX levels were determined by flow cytometry. Figure 4 Both bitopertin and PF-03463275 were shown to demonstrate dose-dependent inhibition of PPIX accumulation by flow cytometry, up to 50%. Bitopertin exhibited an EC50 of 7 nM, and PF-03463275 exhibited an EC50 of 46 nM. Figure 5 Bitopertin and PF-03463275 showed no negative impact on cell viability. Importantly, the LC / MS / MS method showed that bitopertin reduced 5-aminolevulinic acid (5-ALA) and PPIX levels in the EPP K562 cell model with minimal effect on heme formation ( Figure 6 、 7 and 8).

[0589] Another GlyT1 inhibitor also showed dose-dependent inhibition of PPIX accumulation, while the GlyT2 inhibitor ORG-25543, No inhibition was shown at the highest tested concentration of 10 μM (Table 3).

[0590] Table 3: EC values of test compounds in EPP cell model 50

[0591]

[0592]

[0593] Example 8: GlyT1 inhibitors effectively reduce the expression of FECH in cells transduced with lentivirus expressing small interfering RNA (shRNA) PPIX levels in human hematopoietic stem cells

[0594] To investigate the effects of GlyT1 inhibitors in human hematopoietic stem cells with EPP phenotype, lentiviral vectors expressing shRNA of FECH were constructed (Table 4) and transduced into human umbilical cord blood CD34+ cells purchased from Stemexpress at an MOI of 25.

[0595] Table 4: Oligonucleotides used to construct lentiviral vectors carrying shRNA sequences targeting FECH

[0596]

[0597] RT-qPCR of the resulting CD34+ cells showed a 60% decrease in FECH mRNA levels relative to cells treated with a control lentiviral vector ( Figure 9 ). Transduced CD34+ cells were differentiated into erythroid cells over 9 days in StemSpan SFEM II medium supplemented with StemSpan Erythroid Expansion Supplement in the presence of bitopertin (100 nM) or DMSO control. The erythroid cell antigen profile was analyzed using a cytofluorescence strategy employing the following surface markers: CD71 (PE mouse anti-human CD71, BD Biosciencess), glycoprotein A (APC mouse anti-human CD235a, BD Biosciences). After 9 days of differentiation culture, cell viability was greater than 60% in all samples, and more than 80% of cells transduced with lentivirus expressing FECH shRNA showed increased PPIX as determined by flow cytometry ( Figure 10 Biotopertin (100 nM) treatment had no negative effects on erythroid cell surface markers and reduced PPIX accumulation by 60% ( Figure 11 ).

[0598] Although the preferred embodiment of the application has been shown and described herein, it will be apparent to those skilled in the art that this type of embodiment is provided only by way of example. Without departing from the application, those skilled in the art will now expect many variations, changes and replacements. It should be understood that the various alternatives of the application's embodiments as herein described can be used to put the application into practice. It is contemplated that the following claims limit the scope of the application, and thus cover the methods and structures within the scope of these claims and their equivalents.

[0599] In summary, the present invention includes but is not limited to the following:

[0600] 1. A method for treating erythropoietic protoporphyria (EPP), X-linked protoporphyria (XLPP), or congenital erythropoietic porphyria (CEP) in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more glycine transporter 1 (GlyT1) inhibitors or pharmaceutically acceptable salts thereof, or prodrugs of the one or more GlyT1 inhibitors or salts thereof.

[0601] 2. A method for preventing, treating, or reducing the rate of progression and / or severity of one or more complications of EPP, XLPP, or CEP in a subject, the method comprising administering to the subject a pharmaceutical composition comprising one or more GlyT1 inhibitors or pharmaceutically acceptable salts thereof, or prodrugs of the one or more GlyT1 inhibitors or pharmaceutically acceptable salts thereof.

[0602] 3. The method of claim 2, wherein the one or more complications of EPP, XLPP or CEP are selected from the group consisting of acute photosensitivity, cutaneous photosensitivity, edema, erythema, anemia, hypochromic anemia, hemolytic anemia, hemolysis, mild hemolysis, severe hemolysis, chronic hemolysis, hypersplenism, palmar keratosis, bullae, lesions, scars, deformities, nail loss, toe loss, cholestasis, cell lysis, gallstones, cholestatic liver failure, cholelithiasis, mild liver disease, worsening liver disease, end-stage liver disease, erythrodontia, hypercellular bone marrow, myelodysplasia, thrombocytopenia, fetal hydrops and / or intrauterine death.

[0603] 4. A method for preventing or treating EPP, XLPP or CEP in a subject, wherein the method comprises administering to the subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of one or more GlyT1 inhibitors or a pharmaceutically acceptable salt thereof.

[0604] 5. A method for preparing a medicament for treating EPP, XLPP or CEP in a subject, the method comprising administering to the subject at least one GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the one or more GlyT1 inhibitors or a pharmaceutically acceptable salt thereof.

[0605] 6. A method for preparing a medicament for inhibiting protoporphyrin IX (PPIX) synthesis in vivo, the method comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of one or more GlyT1 inhibitors or a pharmaceutically acceptable salt thereof.

[0606] 7. The method of any one of items 1-6, wherein the subject suffers from EPP.

[0607] 8. The method of any one of items 1-6, wherein the subject suffers from XLPP.

[0608] 9. The method of any one of items 1-6, wherein the subject suffers from CEP.

[0609] 10. The method of claim 3, wherein the acute photosensitivity is due to sun exposure.

[0610] 11. The method of any one of items 1-10, wherein the method increases pain-free light exposure in the subject.

[0611] 12. The method of any one of items 1-10, wherein the method reduces the subject's photosensitivity.

[0612] 13. A method of inhibiting PPIX synthesis in vivo, the method comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof.

[0613] 14. A method of inhibiting zinc protoporphyrin IX (ZPPIX) synthesis in vivo, the method comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof.

[0614] 15. A method for inhibiting the synthesis of uroporphyrin I and / or coproporphyrin I in vivo, the method comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof.

[0615] 16. A method of inhibiting 5-aminolevulinic acid (5-ALA) synthesis in vivo, the method comprising administering to a subject a GlyT1 inhibitor or a pharmaceutically acceptable salt thereof, or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof.

[0616] 17. A method according to any one of items 1 to 16, wherein the accumulation of one or more heme intermediates is inhibited, and wherein the one or more heme intermediates are selected from PPIX, ZPPIX, uroporphyrin I, coproporphyrin I and / or 5-ALA.

[0617] 18. A method according to claim 17, wherein the accumulation of one or more heme intermediates is inhibited in a dose-dependent manner.

[0618] 19. The method according to any of the preceding items, wherein the GlyT1 inhibitor exhibits an EC50 of less than 500 nM.

[0619] 20. The method according to any of the preceding items, wherein the GlyT1 inhibitor exhibits an EC50 of less than 100 nM.

[0620] 21. The method according to any of the preceding items, wherein at least 50% of the cell viability is maintained.

[0621] 22. The method according to any of the preceding items, wherein at least 90% of the cell viability is maintained.

[0622] 23. The method of any one of items 1-22, wherein prior to administration of the GlyT1 inhibitor, the subject's PPIX levels are at least 10%, 20%, 30%, 40% or 50% higher than those of healthy subjects.

[0623] 24. The method of any one of items 1-22, wherein prior to administration of the GlyT1 inhibitor, the subject's ZPPIX levels are at least 10%, 20%, 30%, 40% or 50% higher than the ZPPIX levels of healthy subjects.

[0624] 25. The method of any one of items 1-22, wherein the subject has an increased ratio of ZPPIX to free protoporphyrin IX (ZPPIX / PPIX ratio) compared to a subject with EPP.

[0625] 26. The method of any one of items 1-22, wherein prior to administration of the GlyT1 inhibitor, the subject's uroporphyrin I and / or coproporphyrin I levels are at least 10%, 20%, 30%, 40% or 50% higher than the uroporphyrin I and / or coproporphyrin I levels of healthy subjects.

[0626] 27. The method of any one of items 1-22, wherein prior to administration of the GlyT1 inhibitor, the subject's 5-ALA levels are at least 10%, 20%, 30%, 40% or 50% higher than those of healthy subjects.

[0627] 28. The method of any one of items 1-27, wherein the subject's PPIX levels are reduced while the patient's hemoglobin levels are significantly maintained.

[0628] 29. A method according to any one of items 1-28, wherein the patient's PPIX level is reduced by at least 50% (e.g., 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 100%) and the patient's hemoglobin level is reduced by no more than 10% (e.g., 10%, 15%, 20%, 25% and 30%).

[0629] 30. The method of item 28 or 29, wherein the patient's PPIX level is reduced by at least 85% and the patient's hemoglobin level is reduced by no more than 15%.

[0630] 31. The method of any one of items 1-29, wherein hemoglobin levels are reduced by no more than 10% (e.g., 10%, 15%, 20%, 25%, and 30%).

[0631] 32. The method of any one of items 1-31, wherein the dose of the pharmaceutical composition does not cause a significant decrease in hemoglobin levels.

[0632] 33. The method of any one of items 1-8, 10-15, 17-25, and 27-32, wherein the subject has increased levels of free protoporphyrin IX in erythrocytes.

[0633] 34. The method of any one of items 1-8, 10-15, 17-25, and 27-33, wherein the method reduces the level of free protoporphyrin IX in the subject.

[0634] 35. The method of any one of items 1-8, 10-15, 17-25, and 27-34, wherein the method reduces the subject's free protoporphyrin IX level by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0635] 36. The method of any one of items 1-8, 10-15, 17-25, and 27-35, wherein the subject has increased levels of protoporphyrin IX in feces.

[0636] 37. The method of any one of items 1-8, 10-15, 17-25, and 27-36, wherein the method reduces protoporphyrin IX levels in the feces of the subject.

[0637] 38. The method of any one of items 1-8, 10-15, 17-25, and 27-37, wherein the method reduces the level of protoporphyrin IX in the subject's stool by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0638] 39. The method of any one of items 1-38, wherein the subject's plasma porphyrins fluoresce at a peak at 634 nm when illuminated with blue light (e.g., 400-420 nm light).

[0639] 40. The method of any one of items 1-39, wherein the subject's plasma porphyrins fluoresce at a peak between 626 nm and 634 nm when illuminated with blue light (e.g., 400-420 nm light).

[0640] 41. The method of any one of items 1-38, wherein the subject's skin porphyrins fluoresce at a peak at 632 nm when illuminated with blue light (e.g., 400-420 nm light).

[0641] 42. The method of any one of items 1-38, wherein the subject's skin porphyrins fluoresce at a peak between 626 nm and 634 nm when illuminated with blue light (e.g., 400-420 nm light).

[0642] 43. The method of any one of items 1-8, 10-15, 17-25, and 27-42, wherein the subject has increased levels of protoporphyrin IX in the skin.

[0643] 44. The method of any one of items 1-8, 10-15, 17-25, and 27-43, wherein the method reduces protoporphyrin IX levels in the subject's skin.

[0644] 45. The method of any one of items 1-8, 10-15, 17-25, and 27-44, wherein the method reduces the level of protoporphyrin IX in the subject's skin by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0645] 46. The method of any one of items 1-8, 10-15, 17-25, and 27-45, wherein the subject has a protoporphyrin IX level in the skin greater than 0.2 Fluorescent Units (FDU).

[0646] 47. The method of any one of items 1-8, 10-15, 17-25, and 27-45, wherein the subject has a protoporphyrin IX level in the skin greater than 1.0 FDU.

[0647] 48. The method of any one of items 1-8, 10-15, 17-25, and 27-45, wherein the protoporphyrin IX level in the subject's skin is between 1.0 FDU and 2.5 FDU.

[0648] 49. The method of any one of items 1-8, 10-15, 17-25, and 27-45, wherein the subject has a protoporphyrin IX level in the skin greater than 2.5 FDU.

[0649] 50. The method of any one of items 1-8, 10-15, 17-25, and 27-49, wherein the method reduces the level of protoporphyrin IX in the subject's skin to less than 0.5 FDU.

[0650] 51. The method of any one of items 1-8, 10-15, 17-25, and 27-49, wherein the method reduces the level of protoporphyrin IX in the subject's skin to less than 1.0 FDU.

[0651] 52. The method of any one of items 1-8, 10-15, 17-25, and 27-49, wherein the method reduces the level of protoporphyrin IX in the subject's skin to less than 1.5 FDU.

[0652] 53. The method of any one of items 1-8, 10-15, 17-25, and 27-49, wherein the method reduces the level of protoporphyrin IX in the subject's skin to less than 2.0 FDU.

[0653] 54. The method of any one of items 1-8, 10-15, 17-25, and 27-49, wherein the method reduces the level of protoporphyrin IX in the subject's skin to less than 2.5 FDU.

[0654] 55. The method of any one of items 1-8, 10-15, 17-25, and 27-54, wherein the subject has increased levels of protoporphyrin IX in erythrocytes.

[0655] 56. The method of any one of items 1-8, 10-15, 17-25, and 27-55, wherein the method reduces protoporphyrin IX levels in the subject's red blood cells.

[0656] 57. The method of any one of items 1-8, 10-15, 17-25, and 27-56, wherein the method reduces the level of protoporphyrin IX in the subject's red blood cells by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0657] 58. The method of any one of items 1-8, 10-15, 17-25, and 27-57, wherein the subject has a protoporphyrin IX level in erythrocytes greater than 31 μmol L -1 .

[0658] 59. The method of any one of items 1-8, 10-15, 17-25, and 27-58, wherein the protoporphyrin IX level in the erythrocytes of the subject is between 31 μmol L -1 With 53 μmol L -1 between.

[0659] 60. The method of any one of items 1-8, 10-15, 17-25, and 27-58, wherein the subject has a protoporphyrin IX level in erythrocytes greater than 53 μmol L -1 .

[0660] 61. The method of any one of items 1-8, 10-15, 17-25, and 27-58, wherein the method reduces the level of protoporphyrin IX in the subject's red blood cells to less than 53 μmol L -1 level.

[0661] 62. The method of any one of items 1-8, 10-15, 17-25, and 27-58, wherein the method reduces the level of protoporphyrin IX in the subject's red blood cells to less than 31 μmol L -1 level.

[0662] 63. The method of any one of items 1-8, 10-15, 17-25, and 27-58, wherein the method reduces the level of protoporphyrin IX in the subject's red blood cells to less than 15 μmol L -1 level.

[0663] 64. The method of any one of items 1-7, 10-14, 17-25, and 27-63, wherein the subject's ferrochelatase activity level is reduced to between 10% and 35% of the ferrochelatase activity level observed in normal subjects.

[0664] 65. The method of any one of items 1-7, 10-14, 17-25 and 27-64, wherein the subject's ferrochelatase activity level is reduced to less than 50% of the ferrochelatase activity level observed in a normal subject.

[0665] 66. The method of any one of items 1-6, 8, 10-15, 17-25, and 27-63, wherein the subject has a gain-of-function mutation in ALAS2.

[0666] 67. The method of any one of items 1-6, 8, 10-15, 17-25, 27-63, and 66, wherein the subject has increased ALAS2 enzyme activity.

[0667] 68. The method of any one of items 1-6, 8, 10-15, 17-25, 27-63, 66, and 67, wherein the subject has increased levels of zinc protoporphyrin IX in red blood cells.

[0668] 69. The method of any one of items 1-6, 8, 10-15, 17-25, 27-63, and 66-68, wherein the method reduces zinc protoporphyrin IX levels in the subject's red blood cells.

[0669] 70. The method of any one of items 1-6, 8, 10-15, 17-25, 27-63, and 66-69, wherein the method reduces the level of zinc protoporphyrin IX in the subject's red blood cells by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0670] 71. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, and 39-42, wherein the subject has reduced uroporphyrinogen III synthase activity.

[0671] 72. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71, wherein the subject has increased levels of uroporphyrin I and / or coproporphyrin I.

[0672] 73. The method of claim 72, wherein increased levels of uroporphyrin I and / or coproporphyrin I are measured in the urine or red blood cells of the subject.

[0673] 74. The method of claim 72, wherein increased levels of coproporphyrin I are measured in the feces of the subject.

[0674] 75. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-74, wherein the method reduces uroporphyrin I and / or coproporphyrin I levels in the subject.

[0675] 76. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-74, wherein the method reduces urinary porphyrin I levels in the subject.

[0676] 77. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-76, wherein the method reduces the subject's uroporphyrin I level by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0677] 78. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-77, wherein the method reduces coproporphyrin I levels in the subject.

[0678] 79. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-78, wherein the method reduces the subject's coproporphyrin I level by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or at least 100%).

[0679] 80. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-79, wherein the subject has a mutation in UROS.

[0680] 81. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-80, wherein the subject has a genetic defect in the GATA-1 erythroid-specific transcription factor.

[0681] 82. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-81, wherein the subject has red fluorescent urine.

[0682] 83. The method of any one of items 1-5, 9-13, 16-23, 26, 27, 31, 32, 39-42, and 71-82, wherein the subject has a peak between 615 nm and 620 nm using plasma porphyrin fluorescence analysis.

[0683] 84. The method of any one of items 1-83, wherein the subject has a liver disease associated with EPP, XLPP or CEP.

[0684] 85. A method according to any one of items 1-75, wherein the liver disease associated with EPP, XLPP or CEP is cholelithiasis.

[0685] 86. A method according to any one of items 1-75, wherein the liver disease associated with EPP, XLPP or CEP is a mild liver disease.

[0686] 87. A method according to any one of items 1-75, wherein the liver disease associated with EPP, XLPP or CEP is an exacerbated liver disease.

[0687] 88. A method according to any one of items 1-75, wherein the liver disease associated with EPP, XLPP or CEP is end-stage liver disease.

[0688] 89. The method of any one of items 1-88, further comprising administering to the subject an additional active agent and / or supportive therapy.

[0689] 90. The method of item 89, wherein the additional active agent and / or supportive therapy is selected from the group consisting of: sun avoidance, topical sunscreen, skin protection, UVB phototherapy, afamelanotide Bortezomib, proteasome inhibitors, chemical chaperones, cholestyramine, activated charcoal, iron supplementation, liver transplantation, bone marrow transplantation, splenectomy, and blood transfusion.

[0690] 91. The method according to any one of items 1-90, wherein the GlyT1 inhibitor is a compound having the formula

[0691]

[0692] in:

[0693] Ar is unsubstituted or substituted aryl or 6-membered heteroaryl containing 1, 2 or 3 nitrogen atoms, wherein the substituted aryl and the substituted heteroaryl are substituted by one or more substituents selected from the group consisting of hydroxy, halogen, NO2, CN, (C1-C6)-alkyl, (C1-C6)-alkyl substituted by halogen, (C1-C6)-alkyl substituted by hydroxy, (CH2)n-(C1-C6)-alkoxy, (C1-C6)-alkoxy substituted by halogen, NR 7 R 8 、C(O)R 9 、SO2R 10 and -C(CH3)=NOR 7 , or substituted by a 5-membered aromatic heterocyclic ring containing 1 to 4 heteroatoms selected from N and O, said aromatic heterocyclic ring being optionally substituted by a (C1-C6)-alkyl group; R 1 is hydrogen or (C1-C6)-alkyl;

[0694] R 2 is hydrogen, (C1-C6)-alkyl, (C2-C6)-alkenyl, (C1-C6)-alkyl substituted by halogen, (C1-C6)-alkyl substituted by hydroxy, (CH2)n-(C3-C7)-cycloalkyl, CH(CH3)-(C3-C7)-cycloalkyl, (CH2) n+1 -C(O)-R 9 、(CH2) n+1 -CN, bicyclo[2.2.1]heptyl, (CH2) n+1 -O-(C1-C6)-alkyl, (CH2) n -heterocycloalkyl, (CH2)n -aryl or (CH2) containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur or nitrogen n -5- or 6-membered heteroaryl, wherein aryl, heterocycloalkyl and heteroaryl are unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, (C1-C6)-alkyl and (C1-C6)-alkoxy;

[0695] R 3 、R 4 and R 6 are each independently hydrogen, hydroxy, halogen, (C1-C6)-alkyl, (C1-C6)-alkoxy or O-(C3-C6)-cycloalkyl;

[0696] R 5 NO2, CN, C(O)R 9 or SO2R 10 ;

[0697] R 7 and R 8 are each independently hydrogen or (C1-C6)-alkyl;

[0698] R 9 is hydrogen, (C1-C6)-alkyl, (C1-C6)-alkoxy or NR 7 R 8 ;

[0699] R 10 is (C1-C6)-alkyl, (CH2) optionally substituted by halogen n -(C3-C6)-cycloalkyl, (CH2) n -(C3-C6)-alkoxy, (CH2) n -heterocycloalkyl or NR 7 R 8 ;

[0700] n is 0, 1, or 2;

[0701] or a pharmaceutically acceptable salt thereof, or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0702] 92. The method of claim 91, wherein the GlyT1 inhibitor is a compound having the formula The compound bitopertin or a pharmaceutically acceptable salt thereof or a prodrug of the compound or a pharmaceutically acceptable salt thereof.

[0703] 93. The method of any one of items 1-92, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0704] 94. The method of any one of items 1-93, wherein the subject is a subject in need thereof.

[0705] 95. The method according to any one of items 1-94, wherein the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof or a prodrug of the GlyT1 inhibitor or a pharmaceutically acceptable salt thereof is administered in a therapeutically effective amount.

[0706] Incorporated by Reference

[0707] All references cited in this application and their respective references are hereby incorporated by reference in their entirety, to the extent appropriate to teach additional or alternative details, features and / or technical background.

Claims

1. Use of a GlyT1 inhibitor in the preparation of a medicament for treating erythropoietic protoporphyria (EPP) in a subject, wherein the GlyT1 inhibitor is a compound having the formula The compound bitopertin or a pharmaceutically acceptable salt thereof.

2. The use according to claim 1, wherein the use increases pain-free light exposure in the subject.

3. The use according to claim 1, wherein the use reduces the light sensitivity of the subject.

4. The use according to claim 1, wherein the GlyT1 inhibitor inhibits the accumulation of one or more heme intermediates, and wherein the one or more heme intermediates are selected from PPIX and / or 5-ALA.

5. The use according to claim 4, wherein the accumulation of the one or more heme intermediates is inhibited in a dose-dependent manner. The use according to claim 1 , wherein the GlyT1 inhibitor exhibits an EC50 of less than 500 nM.

7. The use according to claim 1, wherein the GlyT1 inhibitor exhibits an EC50 of less than 100 nM.

8. The use according to claim 4, wherein at least 50% of the cell viability is maintained.

9. The use according to claim 4, wherein at least 90% of the cell viability is maintained.

10. The use according to claim 1, wherein the subject's PPIX level is at least 10% higher than that of a healthy subject before administration of the GlyT1 inhibitor.

11. The use according to claim 1, wherein the subject's PPIX level is at least 20% higher than that of a healthy subject before administration of the GlyT1 inhibitor.

12. The use according to claim 1, wherein the subject's PPIX level is at least 30% higher than that of a healthy subject before administration of the GlyT1 inhibitor.

13. The use of claim 1, wherein prior to administration of the GlyT1 inhibitor, the subject's PPIX level is at least 40% higher than that of a healthy subject.

14. The use according to claim 1, wherein the subject's PPIX level is at least 50% higher than that of a healthy subject before administration of the GlyT1 inhibitor.

15. The use according to claim 1, wherein the subject's PPIX level is reduced while the subject's hemoglobin level is significantly maintained.

16. The use of claim 15, wherein the subject's PPIX level is reduced by at least 50% and the subject's hemoglobin level is reduced by no more than 30%.

17. The use according to claim 1, wherein the subject's plasma porphyrin fluoresces at a peak of 634 nm when illuminated with blue light.

18. The use according to claim 1, wherein the subject's plasma porphyrins fluoresce at a peak between 626 nm and 634 nm when illuminated with blue light.

19. The use of claim 1, wherein the subject has increased levels of protoporphyrin IX in the skin.

20. The use of claim 1, wherein the subject's ferrochelatase activity level is reduced to between 10% and 35% of the ferrochelatase activity level observed in healthy subjects.

21. The use of claim 1, wherein the treatment further comprises administering to the subject an additional active agent and / or supportive therapy.

22. The use according to claim 21, wherein the additional active agent and / or supportive therapy is selected from the group consisting of: sun avoidance, skin protection, UVB phototherapy, afamelanotide, bortezomib, cholestyramine, activated charcoal, iron supplementation, liver transplantation, bone marrow transplantation, splenectomy and blood transfusion.

23. The use according to claim 21, wherein the additional active agent and / or supportive therapy is a topical sunscreen.

24. The use according to any one of claims 1 to 23, wherein the medicament further comprises a pharmaceutically acceptable carrier.

25. The use of claim 16, wherein the subject's PPIX level is reduced by at least 50%.

26. The use of claim 16, wherein the subject's PPIX level is reduced by at least 55%.

27. The use of claim 16, wherein the subject's PPIX level is reduced by at least 60%.

28. The use of claim 16, wherein the subject's PPIX level is reduced by at least 65%.

29. The use of claim 16, wherein the subject's PPIX level is reduced by at least 70%.

30. The use of claim 16, wherein the subject's PPIX level is reduced by at least 75%.

31. The use of claim 16, wherein the subject's PPIX level is reduced by at least 80%.

32. The use of claim 16, wherein the subject's PPIX level is reduced by at least 85%.

33. The use of claim 16, wherein the subject's PPIX level is reduced by at least 90%.

34. The use of claim 16, wherein the subject's PPIX level is reduced by at least 95%.

35. The use of claim 16, wherein the subject's PPIX level is reduced by at least 100%.

36. The use of claim 16, wherein the subject's hemoglobin level is reduced by no more than 10%.

37. The use of claim 16, wherein the subject's hemoglobin level is reduced by no more than 15%.

38. The use of claim 16, wherein the subject's hemoglobin level is reduced by no more than 20%.

39. The use of claim 16, wherein the subject's hemoglobin level is reduced by no more than 25%.

40. The use according to claim 17, wherein the blue light is 400-420 nm light.

41. The use according to claim 18, wherein the blue light is 400-420 nm light.

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