Pyridine inhibitors of glucosylceramide synthase and methods of treatment using them

Abstract

Glucosylceramide synthase inhibitors and compositions containing same are disclosed. Methods of using glucosylceramide synthase inhibitors in the treatment of diseases and conditions that benefit from inhibition of glucosylceramide synthase, such as Gaucher disease and Fabry disease, are also disclosed.

Description

[Technical Field]

[0001] Government funding This invention was made with government support under NS092981 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0002] The present invention relates to pyridine inhibitors of glucosylceramide synthase (GCS) and methods of treating conditions and diseases that benefit from inhibition of GCS. [Background technology]

[0003] Lysosomal storage diseases (LSDs), such as Gaucher disease and Fabry disease, occur when glycolipids accumulate in lysosomes due to impaired lysosomal catabolism. There are two general strategies for treating lysosomal storage diseases. The first strategy involves replacing or repairing defective or absent catabolic enzymes (e.g., recombinant enzyme infusion, chaperone therapy, bone marrow transplantation, or gene therapy) (1). Enzyme replacement therapy has been clinically approved for lysosomal storage diseases with peripheral symptoms, but is limited by the inability of the infused recombinant enzyme to distribute within the CNS and the frequent development of autoantibodies against the protein in patients with null mutations.

[0004] The second strategy involves synthetic inhibition therapy, focusing on the identification of small molecule inhibitors of GCS (2). Various classes of GCS inhibitors have been described, including analogs of iminosugars and D-threo-1-phenyl-2-decanoylamino-3-morpholino-propanol (PDMP) (3). The iminosugar N-butyldeoxynojirimycin (NBDNJ) is limited by its micromolar inhibitory activity and limited specificity for the synthase. This limited specificity is associated with a high level of undesirable effects arising from secondary sites of action unrelated to glycolipid synthesis inhibition. These effects, most notably diarrhea, weight loss, and tremors, limit the approved use of NBDNJ in the United States (4). One advantage of NBDNJ over previously reported PDMP-based homologs is its ability to distribute to the CNS. However, recent studies have raised questions about the ability of NBDNJ to reduce glycolipid levels in the CNS (KMAshe et al. Plos One 6:e21758 (2011)).

[0005] Several GCS inhibitors are disclosed, for example, in U.S. Patent Nos. 5,302,609, 5,472,969, 5,525,616, 5,916,911, 5,945,442, 5,952,370, 6,030,995, 6,051,598, 6,255,336, 6,569,889, 6,610,703, 6,660,794, 6,855,830, and 6,855,830. Nos. 6,916,802, 7,253,185, 7,196,205, 7,615,573, 7,994,198, 10,189,784, 10,202,340, and U.S. Patent Publication Nos. 2008 / 0234324, 2016 / 0280643, 2018 / 0044302, 2018 / 0093981, and 2019 / 0031652. Further GCS inhibitors and treatments are disclosed in WO2003 / 035626, WO2008 / 150486, WO2009 / 117150, WO2010 / 014554, WO2010 / 091104, WO2010 / 091164, WO2012 / 129084, WO2015 / 042397, WO2015 / 065937, WO2017 / 204319, and EP331827.

[0006] A compound structurally related to PDMP is N-((1R,2R)-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)octanamide, also known as Genz-112638, and eliglustat tartrate (5). A phase 2 clinical trial of this drug in type 1 Gaucher disease demonstrated efficacy comparable to or greater than that of recombinant β-glucocerebrosidase, as evidenced by reversal of splenomegaly and hepatomegaly, correction of anemia, and improvement in thrombocytopenia and bone mineral density (6). A phase 3 trial with eliglustat tartrate was published, leading to global approval of eliglustat tartrate for type 1 Gaucher disease (7).

[0007] GSC inhibition also holds promise for treating six other lysosomal storage diseases with CNS involvement, including early- and late-onset Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis, and types 2 and 3 Gaucher disease. For example, experimental models of genetic epistasis have shown significantly improved survival in a mouse model of Sandhoff disease that also lacks GM2 synthase (8). However, drug distribution studies suggest that eliglustat tartrate does not transport across the blood-brain barrier (BBB) (5). A possible reason for the poor brain distribution of eliglustat tartrate may be that the drug is a substrate for the p-glycoprotein (MDR1) transporter, resulting in drug efflux (Larsen et al., J Lipid Res 2012, 53, 282-91). The lack of brain penetration offers a clinical advantage for diseases without neurological involvement. These include type 1 Gaucher disease and Fabry disease, the two most common lysosomal storage diseases. Miglustat is approved for type 1 Gaucher disease, but is an oral drug limited to patients intolerant to enzyme replacement therapy. Although it has demonstrated brain penetration, it suffers from several toxicities due to its off-target activity, including neurological activity, such as tremors. These toxicities have not been observed in patients treated with eliglustat (Orphanet et al., J Rare Dis 2019, 14, 128).

[0008] Compounds that inhibit GCS have the potential to treat conditions associated with glycolipid accumulation. However, many GCS inhibitors are limited by poor CNS penetration and / or low activity. A significant advance in the art would be the discovery of GCS inhibitors, particularly those that can cross the BBB and are useful for treating diseases in which GCS inhibition would be beneficial, such as type II or III Gaucher disease, Tay-Sachs disease, Sandhoff disease, diabetes, lupus, and other diseases and conditions associated with glycolipid accumulation in lysosomes. Thus, there remains a need in the art for effective compounds, compositions, and methods useful for treating such diseases, either alone or in conjunction with other therapies used to treat these diseases and conditions. The present invention is directed to meeting this need. Summary of the Invention

[0009] The present invention relates to inhibitors of GCS, methods for preparing GCS inhibitors, compositions comprising the inhibitors, and methods of using the inhibitors in the therapeutic treatment of conditions and diseases that benefit from inhibition of GCS. The compounds of the present invention are potent inhibitors of GCS and, in some embodiments, can cross the BBB, and in some embodiments, have low permeability across the BBB.

[0010] More specifically, the present invention provides a compound having structural formula (I): [ka]

[0011] wherein each X is independently CH, CR 1 , or N, except that exactly one X must be N, A is CH2 or O, R 1 is H, halogen, or OR 5 and R 2 is H, C1-C5 alkyl, or C3-C6 cycloalkyl; R 3 is H or CH3, R 4 is H or F, R 5 is C1-C4 alkyl or C3-C6 cycloalkyl optionally substituted with one or more F; R 6 and R 7 are independently H or C1-C3 alkyl, or R 6 and R 7 taken together with the nitrogen atom to which they are attached form a C4-C8 heterocycloalkyl or heterobicycloalkyl ring, optionally substituted with F or CH3, or a pharmaceutically acceptable salt thereof.

[0012] In one embodiment, the present invention provides a method for treating a condition or disease of interest by administering to an individual in need thereof a therapeutically effective amount of a compound of structural formula (I). Diseases or conditions, such as Gaucher disease, Fabry disease, Sandhoff disease, Tay-Sachs disease, and Parkinson's disease, are treatable by inhibition of the GCS.

[0013] In yet another embodiment, the present invention provides a method of treating a subject having type 2 diabetes, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0014] Also included in the present invention is a method for treating a subject having renal hypertrophy or hyperplasia, hyperplasia associated with diabetic nephropathy, or autosomal dominant polycystic kidney disease (ADPKD), or a viral disease, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0015] Also included in the present invention is a method for reducing plasma TNF-α in a subject in need thereof. Indications treatable by compounds of structural formula (I) include, but are not limited to, multiple myeloma (due to glucosylsphinogosine depletion) and the inhibition of viral and bacterial diseases, including hemolytic uremic syndrome, in which Shiga toxin receptor depletion, i.e., globotriaosylceramide, can be depleted from vascular endothelial cells. The method comprises administering a therapeutically effective amount of a compound of structural formula (I) to a subject.

[0016] Also included in the present invention is a method for lowering blood glucose levels in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0017] Also included in the present invention is a method for reducing glycated hemoglobin levels in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0018] Also included in the present invention is a method for inhibiting glucosylceramide synthase or reducing glycosphingolipid levels in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0019] The present invention also relates to a method for treating a glomerular disease selected from the group consisting of mesangial proliferative glomerulonephritis, collapsing glomerulopathy, proliferative lupus nephritis, crescentic glomerulonephritis, and membranous nephropathy in a subject, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0020] In another embodiment, the present invention relates to a method of treating lupus in a subject, comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0021] In yet another embodiment, in treating the diseases disclosed above, the compounds of structural formula (I) can be administered as the sole therapeutic agent or in combination with a second therapeutic agent known to treat the disease of interest.

[0022] Another embodiment of the present invention is to provide a composition comprising (a) a GCS inhibitor of structural formula (I) and (b) an excipient and / or pharmaceutically acceptable carrier useful in the treatment of a disease or condition benefiting from inhibition of GCS.

[0023] Another embodiment of the present invention is the use of a composition comprising a compound of structural formula (I) and a second therapeutically active agent in a method of treating an individual for a disease or condition in which inhibition of GCS would be beneficial.

[0024] In a further embodiment, the invention provides the use of a composition comprising a GCS inhibitor of structural formula (I) and an optional second therapeutic agent for the manufacture of a medicament for treating a disease or condition of interest, e.g., Gaucher disease or Fabry disease.

[0025] Yet another embodiment of the present invention provides a kit for pharmaceutical use in humans, comprising: (a) a container; (b1) a packaged composition comprising a GCS inhibitor of structural formula (I); and, optionally, (b2) a packaged composition comprising a second therapeutic agent useful in treating a disease or condition of interest; and (c) a package insert containing directions for the use of one or more compositions administered simultaneously or sequentially in treating the disease or condition.

[0026] The GCS inhibitor of structural formula (I) and the second therapeutic agent can be administered together in a single unit dose or separately in multiple unit doses, with the GCS inhibitor of structural formula (I) being administered before the second therapeutic agent, or vice versa. It is contemplated that more than one dose of the GCS inhibitor of structural formula (I) and / or more than one dose of the second therapeutic agent can be administered.

[0027] In one embodiment, the GCS inhibitor of structural formula (I) and the second therapeutic agent are administered simultaneously. In a related embodiment, the GCS inhibitor of structural formula (I) and the second therapeutic agent are administered from a single composition or from separate compositions. In a further embodiment, the GCS inhibitor of structural formula (I) and the second therapeutic agent are administered sequentially. When used in the present invention, the GCS inhibitor of structural formula (I) may be administered in an amount of about 0.005 to about 500 milligrams per dose, about 0.05 to about 250 milligrams per dose, or about 0.5 to about 100 milligrams per dose.

[0028] The compounds of the present invention inhibit GCS and are useful research tools for the in vitro study of GCS and its role in biological processes.

[0029] These and other novel aspects of the present invention will become apparent from the following detailed description of embodiments of the invention. DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention will be described in connection with preferred embodiments. However, it should be understood that the present invention is not limited to the disclosed embodiments. It should be understood that various modifications may be made by those skilled in the art in light of the description of the embodiments of the present invention herein. Such modifications are intended to be encompassed within the scope of the following claims.

[0031] As used herein, the term "GCS" means glucosylceramide synthase.

[0032] The term "diseases or conditions that would benefit from inhibition of GCS" refers to conditions in which GCS and / or the action of GCS is important or necessary, for example, for the onset, progression, or manifestation of the disease or condition, or diseases or conditions that are known to be treated by GCS inhibitors (such as eliglustat tartrate). Examples of such conditions include, but are not limited to, Gaucher disease and Fabry disease. One skilled in the art can readily determine whether a compound treats a disease or condition mediated by GCS, for example, by assays that can be conveniently used to assess the activity of a particular compound.

[0033] The term "second therapeutic agent" refers to a therapeutic agent known to treat the disease or condition of interest that is different from the GCS inhibitor of structural formula (I). In particular, a GCS inhibitor of structural formula (I) can be used to act synergistically with a second therapeutic agent that increases the activity of beta-glucocerebrosidase. For example, if Gaucher disease is the disease or condition of interest, the second therapeutic agent can be known for treating type 1 Gaucher disease or Fabry disease, such as isofagamine, enzyme replacement therapy, or gene therapy.

[0034] The term "disease" or "condition" refers to a disorder and / or abnormality that is primarily considered a pathological state or function and may manifest itself in the form of specific signs, symptoms, and / or dysfunction. As shown below, the compounds of structural formula (I) are inhibitors of GCS and can be used to treat diseases and conditions in which inhibition of GCS is beneficial.

[0035] As used herein, the terms "treat," "treating," "treatment," and the like refer to eliminating, alleviating, or ameliorating a disease or condition and / or its associated symptoms. Treating a disease or condition does not require that the disease, condition, or its associated symptoms be completely eliminated, although this is not entirely impossible. As used herein, the terms "treat," "treating," "treatment," and the like may also include "prophylactic treatment," which refers to reducing the likelihood of a disease or condition recurring, or the likelihood of a previously controlled disease or condition recurring, in a subject who does not have the disease or condition but who is at risk of or susceptible to recurrence or recurrence of the disease or condition. The term "treat" and cognates contemplate administering a therapeutically effective amount of a compound of the invention to an individual in need of such treatment.

[0036] In the sense of the present invention, "treatment" also includes the prevention of relapses or disease phases, as well as the treatment of acute or chronic signs, symptoms, and / or dysfunctions. Treatment can be considered symptomatic, for example to suppress symptoms. This can be achieved over the short term, can be considered over a medium period, or can be long-term treatment, for example as part of maintenance therapy.

[0037] As used herein, the term "therapeutically effective amount" or "effective dose" refers to an amount of an active agent that, when administered by the methods of the present invention, is sufficient to effectively deliver the active agent for the treatment of a condition or disease of interest to an individual in need thereof. In the case of lysosomal storage disorders, a therapeutically effective amount of an agent may reduce (i.e., slow to some extent, and preferably stop) the accumulation of unwanted glycolipids and / or alleviate to some extent one or more of the symptoms associated with the disorder.

[0038] The term "container" means any receptacle and its closure suitable for storing, shipping, dispensing, and / or handling a pharmaceutical product.

[0039] The term "package insert" means the information that accompanies a pharmaceutical product that provides instructions on how to administer the product, along with the safety and effectiveness data necessary to enable physicians, pharmacists, and patients to make informed decisions regarding the use of the product. The package insert is commonly considered the "label" of the pharmaceutical product.

[0040] "Concurrent administration," "administered in combination," "co-administration," and similar phrases refer to the simultaneous administration of two or more agents to a subject being treated. "Concurrently" means that each agent is administered simultaneously or sequentially in any order at different times. However, if not administered simultaneously, it means that they are administered to an individual sequentially and closely enough in time to produce the desired therapeutic effect and act simultaneously. For example, a GCS inhibitor of structural formula (I) can be administered simultaneously with a second therapeutic agent or sequentially in any order at different times. The GCS inhibitor of the present invention and the second therapeutic agent can be administered separately in any suitable form and by any suitable route. It should be understood that if the GCS inhibitor of the present invention and the second therapeutic agent are not administered concurrently, they can be administered to a subject in need thereof in any order.

[0041] For example, a GCS inhibitor of the present invention can be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concurrently with, or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) administration of a second therapeutic agent therapy (e.g., radiation therapy) to an individual in need thereof. In various embodiments, the GCS inhibitor of structural formula (I) and the second therapeutic agent are administered at 1 minute intervals, 10 minutes intervals, 30 minutes intervals, less than 1 hour intervals, 1 hour intervals, 1 to 2 hours intervals, 2 to 3 hours intervals, 3 to 4 hours intervals, 4 to 5 hours intervals, 5 to 6 hours intervals, 6 to 7 hours intervals, 7 to 8 hours intervals, 8 to 9 hours intervals, 9 to 10 hours intervals, 10 to 11 hours intervals, 11 to 12 hours intervals, less than 24 hours intervals, or less than 48 hours intervals. In one embodiment, the components of the combination therapy are administered at intervals of 1 minute to 24 hours.

[0042] In the context of describing the present invention (particularly in the context of the claims), use of the terms "a," "an," "the," and similar referents should be construed to encompass both the singular and the plural unless otherwise indicated. The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated herein as if it were individually recited herein. The use of any and all examples, or exemplary language (e.g., "e.g., "etc."), provided herein is intended to better illustrate the invention and is not a limitation on the scope of the invention unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0043] Compounds that inhibit glycolipid synthesis are known. As such, these compounds can be used to treat diabetes, polycystic kidney disease, Parkinson's disease, susceptible viral diseases, and lysosomal storage diseases such as Tay-Sachs disease, Sandhoff disease, Gaucher disease, and Fabry disease. However, to date, these compounds have been limited by low activity, poor CNS penetration, or both.

[0044] For example, inhibition of glycolipid synthesis is the basis for the treatment of type 1 Gaucher disease with the glucosylceramide (GCS) inhibitor eliglustat tartrate. However, the use of eliglustat for the treatment of glycosphingolipid storage diseases with CNS symptoms is limited by the drug's lack of brain penetration.

[0045] Phase 2 and 3 clinical data for eliglustat tartrate demonstrated clinical responses in type 1 Gaucher disease that were non-inferior to or superior to enzyme replacement therapy, as measured by reductions in spleen and liver volume, correction of anemia, and improvement in thrombocytopenia. Adverse effects observed with NBDNJ, including weight loss, diarrhea, and tremor, were not observed in this clinical trial or extension study. These observations are consistent with eliglustat tartrate's high specificity and lack of CNS penetration, which may be advantageous for glycosphingolipid metabolism disorders without CNS symptoms, including type 1 Gaucher disease and Fabry disease. Some compounds of Formula 1 exhibit poor permeability through the BBB, giving them similar advantages for the treatment of Gaucher disease and Fabry disease. Conversely, the identification of compounds of structural formula (I) that cross the BBB could be of therapeutic benefit for disorders with CNS symptoms, such as GM2 gangliosidosis, Tay-Sachs disease, Sandhoff disease, and type 2 and type 3 Gaucher disease.

[0046] The GCS inhibitors of the present invention are novel and potent inhibitors of GCS and are therefore useful in treating diseases and conditions resulting from the unwanted accumulation of glycolipids, including Gaucher disease and type II diabetes. Also provided is a method for treating a subject having unwanted accumulation of glycolipids, comprising administering a therapeutically effective amount of a compound of the present invention to a subject in need of such treatment.

[0047] Also provided are methods for preventing the growth of undesired glycolipid accumulation in a subject, comprising administering to a subject at risk of developing a condition characterized by undesired glycolipid accumulation a therapeutically effective amount of a compound of structural formula (I). In some embodiments, compounds of structural formula (I) are capable of crossing the BBB and are therefore useful in treating lysosomal storage diseases that previously could not be treated with a GCS inhibitor alone, such as Gaucher disease types II and III, Tay-Sachs disease, and Sandhoff disease.

[0048] More specifically, the present invention provides a compound having structural formula (I): [ka]

[0049] wherein each X is independently CH, CR 1 , or N, except that exactly one X must be N, A is CH2 or O, R 1 is H, halogen, or OR 5 and R 2 is H, C1-C5 alkyl, or C3-C6 cycloalkyl; R 3 is H or CH3, R 4 is H or F, R 5 is C1-C4 alkyl or C3-C6 cycloalkyl optionally substituted with one or more F; R 6 and R7 are independently H or C1-C3 alkyl, or R 6 and R 7 taken together with the nitrogen atom to which they are attached form a C4-C8 heterocycloalkyl or heterobicycloalkyl ring, optionally substituted with F or CH3, or a pharmaceutically acceptable salt thereof.

[0050] The compounds of structural formula (I) are used in methods for treating diseases or conditions that benefit from inhibition of GCS, such as Gaucher disease, Fabry disease, Tay-Sachs disease, Sandhoff disease, diabetes, hypertrophy or hyperplasia associated with diabetic neuropathy, polycystic kidney disease, viral diseases, lupus, elevated plasma TNF-α levels, elevated glycated hemoglobin levels, and glomerular diseases. The methods involve administering a therapeutically effective amount of a compound of structural formula (I) to an individual in need thereof. The methods also include administering to the individual, in addition to a compound of structural formula (I), a second therapeutic agent, including enzyme replacement therapy. The second therapeutic agent is selected from drugs or biologics known to be useful in treating diseases or conditions afflicting individuals in need thereof. Such diseases and conditions include, but are not limited to, Parkinson's disease, multiple myeloma, and ADPKD.

[0051] As used herein, the terms "C1-C5 alkyl" and "C1-C4 alkyl" refer to straight-chain and branched saturated hydrocarbon groups, non-limiting examples of which include methyl, ethyl, straight-chain and branched propyl groups, straight-chain and branched butyl groups, and straight-chain and branched pentyl groups.

[0052] The terms "C3-C6 cycloalkyl," "C4-C8 heterocycloalkyl or heterobicycloalkyl" refer to saturated cycloalkyl and heterocycloalkyl groups containing from 4 to 8 atoms, including, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, and corresponding rings containing a nitrogen atom.

[0053] The alkyl, cycloalkyl, and heterocycloalkyl groups may be substituted with one or more F.

[0054] In various embodiments, [ka] teeth, [ka] wherein X is CH or CR 1 is.

[0055] In one preferred embodiment, [ka] teeth [ka] and In the formula, X is CH or CR 1 is.

[0056] In various embodiments, the pyridine ring system is CR 1 If it contains, R 1 is —H, —OC(CH3)3, —OCH3, —OCF3, —OCH2CF3, or Cl.

[0057] In a preferred embodiment, R 2 is H or CH3.

[0058] In another preferred embodiment, —NR 6 R 7 teeth, [ka] is.

[0059] In various embodiments, [ka] teeth, [ka] is.

[0060] The present invention further includes all possible stereoisomers of the compound of structural formula (I). The present invention includes both racemates and optically active isomers. When a compound of structural formula (I) is desired as a single enantiomer, it can be obtained by resolution of the final product or by stereospecific synthesis from isomerically pure starting materials or by use of chiral auxiliary reagents (see, for example, Z. Ma et al., Tetrahedron: Asymmetry, 8(6), pp. 883-888 (1997)). Resolution of the final product, intermediate, or starting material can be achieved by any suitable method known in the art. Furthermore, in situations where tautomers of a compound of structural formula (I) are possible, the present invention is intended to include all tautomeric forms of the compound.

[0061] Salts of the compounds of the present invention are also encompassed by the present invention and can be used in the methods disclosed herein. Pharmaceutically acceptable salts of the compounds of the present invention are often preferred in the methods of the present invention. As used herein, the term "pharmaceutically acceptable salt" refers to a salt or zwitterionic form of a compound of structural formula (I). Salts of compounds of formula (I) can be prepared during the final isolation and purification of the compound, or can be prepared separately by reacting the compound with an acid having a suitable cation. A pharmaceutically acceptable salt of a compound of structural formula (I) can be an acid addition salt formed using a pharmaceutically acceptable acid. Examples of acids that can be used to form pharmaceutically acceptable salts include inorganic acids such as nitric acid, boric acid, hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, and organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid. Non-limiting examples of salts of the compounds of the present invention include hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, 2-hydroxyethanesulfonate, phosphate, hydrogenphosphate, acetate, adipate, alginate, aspartate, benzoate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerolphosphate, hemisulfate, heptanoate, formate, succinate, fumarate, maleate, ascorbate, isethionate, salicylate, methanesulfonate, mesitylenesulfonate, sodium phosphate ... These include, but are not limited to, phthalenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylproprionate, picrate, pivalate, propionate, trichloroacetate, trifluoroacetate, phosphate, glutamate, glutarate, bicarbonate, paratoluenesulfonate, undecanoate, lactate, citrate, tartrate, gluconate, methanesulfonate, ethanedisulfonate, benzenesulfonate, and p-toluenesulfonate.In addition, available amino groups present in the compounds of the invention may be quaternized with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and steryl chlorides, bromides, and iodides, and benzyl and phenethyl bromides. In view of the above, any reference to a compound of the invention presented herein is intended to include the compound of structural formula (I), as well as pharmaceutically acceptable salts, hydrates, or solvates thereof.

[0062] Some specific embodiments of the present invention include, but are not limited to, the following. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0063] Compound synthesis The compounds of the present invention were prepared as follows: The following synthetic scheme is representative of the reactions used to synthesize compounds of structural formula (I). Modifications and alternative schemes for preparing the GCS inhibitors of the present invention are readily within the capabilities of one of ordinary skill in the art.

[0064] Preparation and spectroscopic data of compounds of formula (I) Chemical names follow CAS or IUPAC nomenclature. Starting materials and reagents were purchased from Fisher, Sigma-Aldrich Lancaster, Fluka, or TCI-America and used without purification. All reaction solvents were purchased from Fisher and used as received. Reactions were monitored by TLC using precoated silica gel 60F254 plates or analytical reverse-phase HPLC, as described in the following paragraphs. Silica gel chromatography was performed on silica gel (220-240 mesh) obtained from Silicycle.

[0065] NMR spectra were recorded on a Bruker 500 MHz spectrometer. Chemical shifts were referenced to the internal standard CDCl3: δ = 7.28 ( 1 H NMR (H NMR) is reported in δ (parts per million) referenced to the hydrogen residue of the deuterated solvent. Mass spectra were recorded on a Micromass LCT time-of-flight instrument using positive ion electrospray ionization mode. Compound purity was assessed by analytical reverse-phase HPLC (Agilent Eclipse Plus C18 4.6 x 75 mm column (3.5 μm silica), 254 nm detection) using a gradient of 10-90% CH3CN / water over 6 min.

[0066] Unless otherwise stated, all temperatures are in degrees Celsius.

[0067] In these examples and elsewhere, the abbreviations have the following meanings: [Table 2-1] [Table 2-2]

[0068] Representative Scheme 1 [ka] (1R,3S,5S)-1,3-bis(6-methoxypyridin-3-yl)-5-phenyltetrahydro-3H,8H-oxazolo[4,3-c][1,4]oxazin-8-one (1-3) (S)-5-Phenylmorpholin-2-one (1-2, 2.2 g, 12.42 mmol) and 6-methoxy-3-nicotinaldehyde (1-1, 5.11 g, 37.2 mmol) were combined in toluene (60 mL). The flask was equipped with a magnetic stir bar and a Dean-Stark trap. The reaction mixture was heated at reflux under nitrogen for 19 hours (pot temperature 152 °C). It was concentrated in vacuo. Purification was carried out by flash chromatography (EtOAc / hexanes): hexanes to 10% EtOAc / hexanes to 20% to 30%. (1R,3S,5S)-1,3-bis(6-methoxypyridin-3-yl)-5-phenyltetrahydro-3H,8H-oxazolo[4,3-c][1,4]oxazin-8-one (2.7 g, 6.2 mmol, 50.2% yield). The product solidified upon concentration to give a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.23(d,J=2.4Hz,1H), 8.00(d,J=2.3Hz,1H), 7.81(dd,J=8.6,2.5Hz,1H) , 7.60(m,1H), 7.34-7.07(m,5H), 6.89(d,J=8.6Hz,1H), 6.56(d,J=8.5Hz ,1H), 5.42(d,J=6.6Hz,1H), 5.30(s,1H), 4.62-4.42(m,2H), 4.35(dd,J= 11.4,3.7Hz,1H), 4.19(dd,J=10.4,3.7Hz,1H), 3.85(s,3H), 3.72(s,3H). HPLC:t r 7.15 minutes

[0069] (2S,3R)-3-hydroxy-2-(((R)-2-hydroxy-1-phenylethyl)amino)-3-(6-methoxypyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-one (1-4) A solution of (1R,3S,5S)-1,3-bis(6-methoxypyridin-3-yl)-5-phenyltetrahydro-3H,8H-oxazolo[4,3-c][1,4]oxazin-8-one (1-3, 1.4 g, 3.23 mmol) and pyrrolidine (1.6 g, 22.6 mmol) in CHCl (40 mL) was stirred overnight at room temperature. The mixture was concentrated. The mixture was treated with 2 M HCl until acidic. MeOH was added and stirred at 60 °C for 2 h. The mixture was concentrated. The aqueous layer was basified with saturated aqueous NaHCO and partitioned between ether and water. The aqueous layer was extracted (3 × EtOAc). The combined extracts were washed with saturated NaCl solution and dried over MgSO. Purification was performed by flash chromatography (MeOH / CHCl). (3R)-3-Hydroxy-2-(((S)-2-hydroxy-1-phenylethyl)amino)-3-(6-methoxypyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-one was obtained (0.9 g, 2.3 mmol, 72.3% yield). 1 H NMR (400MHz, chloroform-d) δ7.43-7.16(m,4H), 7.13(m,1H), 7.00(m,1H), 6.83(t,J=8.5Hz,1H), 4.50(d,J=8.6Hz,1H), 3.83(s,3H), 3.79 -3.59(m,2H), 3.14-2.95(m,2H), 2.95-2.76(m,2H), 2.95-2.22(m,1H), 2.04-1.68(m,2H), 2.51-0.99(m,4H)HR-MS(ESI)m / z:[M+H+Na] + Calculated for 408.190; Found: 408.189. HPLC: t r 4.33 minutes

[0070] (1R,2R)-2-(((R)-2-hydroxy-1-phenylethyl)amino)-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-1-ol (1-5) To a solution of (2S,3R)-3-hydroxy-2-(((R)-2-hydroxy-1-phenylethyl)amino)-3-(6-methoxypyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-one (1-4, 1 g, 2.3 mmol) in dry THF (20 mL) was added lithium aluminum(III) hydride (2.94 mL, 7.07 mmol). The resulting mixture was stirred at room temperature overnight. It was heated to 50° C. for 1 h. It was cooled to 0° C. and treated with 0.15 mL of water, followed by 0.15 mL of 15% NaOH, and then 0.45 mL of water. It was stirred for 60 min and the solid material was filtered through Celite. It was concentrated. It was treated with 0.2 mL of 10% citric acid. It was placed under high vacuum overnight. Purification by flash chromatography using 2% MeOH / CHCl to 5%, then 7% ammonia / methanol / CHCl from 2% to 5%, then 10%. Concentration of the appropriate fractions gave 2-(((S)-2-hydroxy-1-phenylethyl)amino)-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-1-ol (0.45 g, 1.211 mmol, 46.7% yield). 1 H NMR (400MHz, chloroform-d) δ8.10(dt,J=2.5,0.7Hz,1H), 7.56(m,1H), 7.40-7.1 4(m,4H), 6.73(dd,J=8.5,0.7Hz,1H), 4.52(d,J=5.2Hz,1H), 3.93(s,3H),3. 79(m,1H), 3.62(dd,J=11.0,4.4Hz,1H), 3.57-3.40(m,1H), 2.97(m,1H), 2.5 5(dd,J=12.5,8.2Hz,2H), 2.37(m,3H), 2.30-2.04(m,1H), 1.89-1.31(m,4H). HR-MS(ESI)m / z:[M+H] + Calculated for 372.221; Found: 372.225. HPLC: t r 4.19 minutes

[0071] (1R,2R)-2-amino-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-1-ol (1-6) To (1R,2R)-2-(((R)-2-hydroxy-1-phenylethyl)amino)-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-1-ol (1-5, 0.45 g, 1.2 mmol) in MeOH (15 mL) and 2 M HCl (5 mL) was added 10% Pd / C Degussa (25 mg). Nitrogen was bubbled through the solution for 5 minutes, and the vessel was placed on a Parr hydrogenation apparatus. The solution was placed under vacuum and then filled with H2 gas. The mixture was stirred under hydrogen overnight. The vessel was placed under vacuum and the H2 gas was removed. The mixture was filtered over Celite with MeOH eluent. Concentrated and used without purification. (0.23 g, 0.915 mmol, 76% yield)

[0072] 1 H NMR (500 MHz, chloroform-d) δ 7.62-7.61 (m, 1H), 7.03-6.71 (m, 2H), 4.59 (m, 1H), 3.79 (m, 2H), 3.14 (m, 1H), 2.75-2.26 (m, 5H), 1.78 (br s, 4H). HPLC: t r 0.95 minutes

[0073] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (1-7, Example 1) To a solution of (1R,2R)-2-amino-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-1-ol (1-6, 0.1 g, 0.39 mmol) in dry DMF was added 1H-benzo[d][1,2,3]triazol-1-ol (0.07 g, 0.52 mmol), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (0.10 g, 0.52 mmol), and Hunig's base. 2-(2,3-Dihydro-1H-inden-2-yl)acetic acid (0.08 g, 0.44 mmol) was added, and the resulting mixture was stirred at room temperature overnight. It was diluted with EtOAc / ether and washed with saturated NaCl. It was purified by flash chromatography (MeOH / CHCl). 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide was obtained (0.07 g, 0.171 mmol, yield 43.0%). 1 H NMR (400MHz, chloroform-d) δ8.19(d,J=2.8Hz,1H), 7.50(d,J=8.7Hz,1H), 7.36-7.21(m,1H), 7.21-6.99(m,3H), 6.91(d,J=8.1Hz, 1H), 4.99(d,J=2.9Hz,1H), 4.55(m,1H), 3.84(s,3H), 2.95(m,2H), 2.85-2.34(m,8H), 2.37-2.07(m,3H), 1.78(d,J=3.3Hz,4H). HR-MS(ESI)m / z:[M+H] + Calculated for 409.237; Found: 409.243. HPLC: t r 4.5 minutes

[0074] Example 2 was prepared as described in Representative Scheme 1 of Example 1 (1S,3S,5S)-1,3-bis(5-methoxypyridin-2-yl)-5-phenyltetrahydro-3H,8H-oxazolo[4,3-c][1,4]oxazin-8-one (S)-5-Phenylmorpholin-2-one (0.52 g, 2.92 mmol) and 5-methoxypicolinaldehyde (1, 7.29 mmol) were added to a flask equipped with a magnetic stir bar and a Dean-Stark trap in toluene (200 mL). The reaction mixture was heated under nitrogen at reflux (pot temperature 152 °C) for 19 h. The mixture was concentrated in vacuo. Purification was carried out by flash chromatography (EtOAc / hexanes) using hexanes to 10% EtOAc / hexanes to 20% to 30% EtOAc / hexanes to give (1S,3S,5S)-1,3-bis(5-methoxypyridin-2-yl)-5-phenyltetrahydro-3H,8H-oxazolo[4,3-c][1,4]oxazin-8-one (0.65 g, 1.50 mmol, 51.4% yield). The product solidified upon concentration to give a white solid. Trituration with ether / hexanes was performed. NMR and HPLC were performed. 1 H NMR (400MHz, chloroform-d) δ8.32(d,J=2.9Hz,1H), 8.01(d,J=2.9Hz,1H), 7.47(d,J=8.6Hz,1H), 7.43-7.07(m,6H) ), 6.98(dd,J=8.6,2.9Hz,1H), 5.70-5.37(m,2H), 4.95(d,J=7.2Hz,1H), 4.63-4.06(m,4H), 4.06-3.52(m,6H). HPLC:t r 6.9 minutes

[0075] (2S,3R)-3-Hydroxy-2-(((R)-2-hydroxy-1-phenylethyl)amino)-3-(5-methoxypyridin-2-yl)-1-(pyrrolidin-1-yl)propan-1-one (1S,3S,5S)-1,3-bis(5-methoxypyridin-2-yl)-5-phenyltetrahydro-3H,8H-oxazolo[4,3-c][1,4]oxazin-8-one (0.65 g, 1.5 mmol) and pyrrolidine (0.64 g, 9.0 mmol) in CHCl (40 mL) were stirred at room temperature overnight. Concentrated. Treated with 2 M HCl until acidic. MeOH was added and stirred at 60 °C for 2 h. Concentrated. The aqueous layer was basified. Partitioned between ether and water. The aqueous layer was extracted (3 × EtOAc). The combined extracts were washed with saturated NaCl solution and dried over MgSO. Purified by flash chromatography (MeOH / CHCl). (3S)-3-Hydroxy-2-(((S)-2-hydroxy-1-phenylethyl)amino)-3-(5-methoxypyridin-2-yl)-1-(pyrrolidin-1-yl)propan-1-one was obtained (0.32 g, 0.83 mmol, 55.4% yield). 1 H NMR (400MHz, chloroform-d) δ8.32-8.18(m,1H), 7.42(d,J=8.6Hz,1H), 7.39-7.03(m,4H), 4.74(d,J=5.2Hz,2H), 4.23-3.95( m,2H), 3.85(d,J=2.2Hz,3H), 3.78-3.40(m,4H), 3.14(m,2H), 3.01(m,1H), 2.83(m,1H), 2.71(m,2H), 1.81-1.34(m,4H). HR-MS(ESI)m / z:[M+H+Na] + Calculated for 409.237; Found: 409.243. HPLC: t r 4.3 minutes

[0076] (1R,2R)-2-(((R)-2-hydroxy-1-phenylethyl)amino)-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol To a 0° C. solution of (2S,3R)-3-hydroxy-2-(((R)-2-hydroxy-1-phenylethyl)amino)-3-(5-methoxypyridin-2-yl)-1-(pyrrolidin-1-yl)propan-1-one (0.34 g, 0.88 mmol) in dry THF (15 mL) was added lithium aluminum(III) hydride (0.067 g, 1.76 mmol). The resulting mixture was stirred at room temperature overnight. It was heated to 50° C. for 1 h. It was cooled to 0° C. and treated with 0.15 mL of water, followed by 0.15 mL of 15% NaOH, then 0.45 mL of water. It was stirred for 60 min and the solid material was filtered through Celite. It was concentrated. It was treated with 0.2 mL of 10% citric acid. It was placed under high vacuum overnight. Purification by flash chromatography using 2% MeOH / CHCl to 5%, then 7% ammonia / methanol / CHCl from 2% to 5%, then 10% afforded (1S)-2-(((S)-2-hydroxy-1-phenylethyl)amino)-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol (0.14 g, 0.38 mmol, 42.7% yield). 1 H NMR (400MHz, chloroform-d) δ8.10(m,1H), 7.56m,1H), 7.39-7.21(m,8H), 6.73(d,J=8.6Hz,1H), 4.52(d,J=5.4Hz,1H), 3.94(s,3H), 3. 79(m,1H), 3.62(d,J=1.0Hz,1H), 3.56-3.42(m,2H), 2.97(m,1H), 2.55(m,1H), 2.37(m,3H), 2.25-2.13(m,1H), 1.76-1.62(m,3H). HR-MS(ESI)m / z:[M+H] + Calculated for 372.221; Found: 372.225. HPLC: t r 4.5 minutes

[0077] (1R,2R)-2-amino-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol To a solution of (1R,2R)-2-(((R)-2-hydroxy-1-phenylethyl)amino)-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol (0.45 g, 1.21 mmol) in MeOH (15 mL) and 2 M HCl was added 10% Pd / C Degussa (25 mg). Nitrogen was bubbled through the solution for 5 minutes and the vessel was placed on a Parr hydrogenation apparatus. The solution was placed under vacuum and then filled with H2 gas. The mixture was stirred under hydrogen overnight. The vessel was placed under vacuum and the H2 gas was removed. The mixture was filtered over Celite with MeOH eluent. Concentrated and used without purification. (1S)-2-Amino-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol (0.08 g, 0.32 mmol, 84%) oil.

[0078] 1 H NMR (400MHz, chloroform-d) δ8.36-7.99(m,1H), 7.58(dd,J=8.5,2.4Hz,1H), 6.74(d,J=8.5Hz,1H ), 4.62(d,J=3.9Hz,1H), 3.93(d,J=0.6Hz,3H), 3.13(m,1H), 2.81-2.33(m,7H), 1.77(m,4H). HPLC:t r 1.0 minutes

[0079] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 2) To a solution of (1R,2R)-2-amino-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol (0.1 g, 0.39 mmol) in dry DMF was added 1H-benzo[d][1,2,3]triazol-1-ol (0.070 g, 0.52 mmol), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (0.10 g, 0.52 mmol), and Hunig's base. 2-(2,3-Dihydro-1H-inden-2-yl)acetic acid (0.08 g, 0.44 mmol) was added, and the resulting mixture was stirred at room temperature overnight. It was diluted with EtOAc / ether and washed with saturated NaCl. It was purified by flash chromatography (MeOH / CHCl). 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(6-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide was obtained (0.05 g, 0.122 mmol, 30.7% yield). 1 H NMR (400MHz, chloroform-d) δ7.89(dd,J=6.3,3.1Hz,1H), 7.44(dd,J=6.3,3.0Hz,1H), 7.38-6.93(m,7H) , 3.34-3.09(m,5H), 3.08-2.77(m,5H), 2.77-2.26(m,7H), 1.42(d,J=6.7Hz,1H), 1.29-1.01(m,5H). HPLC:t r 4.5 minutes

[0080] Representative Scheme 2 [ka]

[0081] Preparation of (R)-benzyl (3-hydroxy-1-(methoxy(methyl)amino)-1-oxopropan-2-yl)carbamate (2-3, step 2-1).

[0082] To a stirred solution of compound 2-1 (5.00 g, 20.9 mmol), compound 2-2 (6.11 g, 62.6 mmol), and DIPEA (18.2 mL, 104 mmol) in anhydrous DMF (50 mL) was added HATU (8.74 g, 23.0 mmol) at room temperature under nitrogen. The resulting mixture was stirred overnight. The reaction mixture was then quenched with water (100 mL) and extracted with EtOAc (3×). The combined extracts were washed with 10% aqueous LiCl (3×), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was further dried under high vacuum to give compound 2-3 (5.24 g, 88%) as a colorless syrup: ESI MS, m / z = 283 [M+H]. + .

[0083] Preparation of (R)-benzyl (1-(methoxy(methyl)amino)-1-oxo-3-(pyrrolidin-1-yl)propan-2-yl)carbamate (2-4, step 2-2).

[0084] To a stirred solution of compound 2-3 (3.50 g, 12.4 mmol) and EtN (5.2 mL, 37.3 mmol) in anhydrous THF (50 mL) was added MsCl (1.4 mL, 18.1 mmol) dropwise under nitrogen at 0 °C. The resulting mixture was stirred at 0 °C for 30 min. LC / MS indicated the reaction was complete. The reaction mixture was then diluted with EtOAc and quenched with water. The layers were separated, and the aqueous layer was back-extracted with EtOAc (3x). The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was further dried under high vacuum and dissolved in anhydrous THF (50 mL). To the resulting solution, KCO (3.43 g, 24.8 mmol), NaI (1.86 g, 12.4 mmol), and pyrrolidine (2.1 mL, 25.2 mmol) were added at room temperature under nitrogen. The reaction mixture was then heated at 50° C. for 1.5 hours. After this time, the reaction mixture was cooled to room temperature and filtered. The filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 2% to 5% MeOH / CHCl to give compound 2-4 (1.57 g, 38%) as a dark red syrup: ESI MS, m / z=336 [M+H] + .

[0085] Preparation of (+ / -)-benzyl (1-(4-chloropyridin-2-yl)-1-oxo-3-(pyrrolidin-1-yl)propan-2-yl)carbamate (2-6, step 2-3).

[0086] To a stirred mixture of compound 2-4 (2.73 g, 8.14 mmol) and compound 2-5 (2.14 g, 8.94 mmol) in anhydrous THF (60 mL) was added 2M HCl in THF (16.3 mL, 32.6 mmol). iThe PrMgCl solution was added dropwise over 60 minutes at room temperature under N2. The resulting mixture was stirred for 60 minutes. After this time, the reaction mixture was quenched with brine and extracted with EtOAc (3x). The combined extracts were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was further dried under high vacuum to give crude compound 2-6 as an orange oil (3.10 g), which was carried on to the next step without further purification: ESI MS, m / z = 388 [M+H] + .

[0087] Preparation of (+ / -)-benzyl (1-(4-chloropyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)carbamate (2-7, step 2-4).

[0088] To a stirred solution of crude compound 2-6 (3.10 g) in anhydrous THF (120 mL) was added dropwise a 1 M solution of L-Selectride in THF (16.0 mL, 16.0 mmol) under N at −78 °C. The resulting mixture was stirred at −78 °C for 2 h, then quenched with saturated aqueous NH4Cl and extracted with EtOAc (3×). The combined extracts were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 5% to 50% (80:18:2 = CHCl2 / MeOH / NH4OH) / CHCl2 to give compound 2-7 as a brown foam (1.50 g, 47% for two steps): ESI MS, m / z = 390 [M+H]. + .

[0089] Preparation of (+ / -)-2-amino-1-(4-chloropyridin-2-yl)-3-(pyrrolidin-1-yl)propan-1-ol hydrobromide (2-8, step 2-5).

[0090] To a stirred solution of compound 2-7 (1.37 g, 3.52 mmol) in HOAc (50 mL) was added a 33% HBr solution in HOAc (8 mL) dropwise over 20 minutes at room temperature. The reaction mixture was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The resulting residue was further dried under high vacuum to give crude compound 2-8 as a brown foam (1.75 g), which was carried on to the next step without further purification: ESI MS, m / z = 256 [M+H] + .

[0091] Preparation of (+ / -)-N-(1-(4-chloropyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (2-10, step 2-6) (Example 23).

[0092] To a stirred solution of crude compound 2-8 (1.75 g), compound 2-9 (618 mg, 3.51 mmol), and DIPEA (6.11 mL, 35.1 mmol) in anhydrous THF (80 mL) was added HATU (1.33 g, 3.50 mmol). The reaction mixture was stirred for 1 h. After this time, the reaction was quenched with brine and extracted with EtOAc (3x). The combined extracts were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 5% to 40% (80:18:2 CHCl / MeOH / NHOH) / CHCl to give compound 2-10 as a white solid (720 mg, 50% over two steps): ESI MS, m / z = 414 [M+H]. + . 1 H NMR(500MHz,DMSO-d6)δ 8.47(d,J=5.0Hz,1H), 7.53(d,J=2.0Hz,1H), 7.48(bs,1H), 7.40(d,J=3.5Hz,1H), 7.15-7.05(m,4H), 5.89 (bs,1H), 4.91(s,1H), 4.34(bs,1H), 2.83(bs,1H), 2.76-2.42(m,7H), 2.37-2.08(m,5H), 1.71(bs,4H);ESI MS, m / z=414[M+H] + .

[0093] Preparation of N-((1S,2R)-1-(4-chloropyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 25, EEF-AN-21, steps 2-7).

[0094] SFC chiral separation of compounds (+ / -)-2-10: Analytical SFC method for compound (+ / -)-2-10: Chiralpak® OD-H (1.6 x 100 mm, 5 microns); Mobile phase: 20% MeOH in CO2; 1.5 ml / min; 8 min at 40°C; 1750 psi; 220 nm; Injection: 4 uL of a 1 mg / ml solution in MeOH, retention time (t r ): 2.94 minutes (Example 25) and 3.57 minutes (Example 26).

[0095] Preparative SFC chiral separation of compound (+ / -)-2-10 (145 mg): ChiralCel® OD-H (30 × 250 mm, 5 microns); mobile phase: 20% MeOH in CO; 100 g / min at 40 °C; 120 bar; 220 nm; injection: 0.7 mL of 0.1 M MeOH solution. After separation from SFC, the resulting residue (after concentration under reduced pressure) containing the desired enantiomer was further purified by flash column chromatography on silica gel eluting with 0% to 80% (80:18:2 CHCl / MeOH / NHOH) / CHCl to give compound 25 (48 mg, 33%) as a white solid: ESI MS, m / z = 414 [M+H]. + .

[0096] Example 25: 1H NMR(300MHz,CDCl3)δ 8.40(d,J=5.4Hz,1H), 7.66(d,J=2.0Hz,1H), 7.25-7.23(m,1H), 7.18-7.11(m,4H), 7.00-6.96(m,1H), 5.02-5 ESI MS m / z 414[M+H] + .

[0097] Example 26: 1 H NMR(300MHz,CDCl3)δ 8.40(d,J=6.0Hz,1H), 7.67(d,J=2.1Hz,1H), 7.25-7.22(m,1H), 7.18-7.10(m,4H), 7.00-6.97(m,1H), 5.02-5 .00(m,1H), 4.64-4.56(m,1H), 3.08-2.91(m,2H), 2.88-2.40(m,9H), 2.36-2.19(m,2H), 1.86-1.73(m,4H); MS m / z 414[M+H] + .

[0098] The following examples were prepared by the synthetic route shown in representative Scheme 2, using the same procedures as described for Examples 23, 25, and 26. Typically, the final products were isolated as racemates, although in some cases the racemates were separated into individual enantiomers by chiral chromatography (as described for Examples 25 and 26).

[0099] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-1-(6-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 3) [ka]

[0100] 1H NMR (500 MHz, DMSO-d6) δ 9.1(s,1H), 7.78(d,J=9.5Hz,1H), 7.71(t,J=7.5Hz,1H), 7.11-7.05(m, 5H), 6.71(d,J=8.5Hz,1H), 4.79-4.77(m,2H), 3.87(s,3H), 3.65-3.50( m,3H), 3.36-3.25(m,1H), 3.16-3.07(m,2H), 2.75-2.70(m,1H), 2.55-2 .45(m,2H), 2.39-2.36(m,1H), 2.18-2.09(m,3H), 2.01-1.80(m,4H);ESI MS, m / z=410 [M+H] + .

[0101] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-1-(pyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 4) [ka]

[0102] 1 H NMR(500MHz,DMSO-d6)δ 8.47(bs,1H), 7.73(dd,J=1.5,7.5Hz,1H), 7.47(d,J=8.0Hz,1H), 7.38(d,J=9.0Hz,1H), 7.23(m,1H), 7.12-7.06(m,4H), 5.68(d,J= ESI MS, m / z=380[M+H] + .

[0103] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-3-(pyrrolidin-1-yl)-1-(5-(trifluoromethoxy)pyridin-2-yl)propan-2-yl)acetamide (Example 5) [ka]

[0104] 1 H NMR(500MHz,DMSO-d6)δ 8.57(d,J=2.8Hz,1H), 7.85(dd,J=1.6,8.6Hz,1H), 7.60(d,J=8.6Hz,1H), 7.46(d,J=9.2Hz,1H), 7.11-7.06(m,4H) ), 5.90(bs,1H), 4.93(d,J=1.8Hz,1H), 4.35-4.28(m,1H), 2.85-2.45(m,7H), 2.40-2.05(m,5H), 1.69(bs,4H);ESI MS, m / z=464[M+H] + .

[0105] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-3-(pyrrolidin-1-yl)-1-(5-(2,2,2-trifluoroethoxy)pyridin-2-yl)propan-2-yl)acetamide (Example 6) [ka]

[0106] 1 H NMR(500MHz,DMSO-d6)δ 8.30(d,J=2.7Hz,1H), 7.50-7.46(m,1H), 7.45-7.41(m,1H), 7.39-7.34(m,1H), 7.13-7.05(m,4H), 5.68-5.64(m,1H), 4.87-4.76(m,3H), ESI M.S. m / z 478[M+H] + .

[0107] N-((1S,2R)-1-(5-(tert-butoxy)pyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 7) [ka]

[0108] 1 H NMR(500MHz,DMSO-d6)δ 8.14(s,1H), 7.38-7.36(m,3H), 7.12-7.11(m,4H), 5.80(bs,1H), 4.85(d,J=2.5Hz,1H), 4.28-4.22(m,1H), 2.78-2.7 4(m,2H), 2.68-2.62(m,1H), 2.54-2.43(m,4H), 2.31-2.29(m,4H), 2.11-2.08(m,2H), 1.69(bs,4H), 1.28(s,9H);ESI MS, m / z=452[M+H] + .

[0109] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(2-methoxypyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 8) [ka]

[0110] 1 H NMR(500MHz,DMSO-d6)δ 8.04(d,J=5.5Hz,1H), 7.50(d,J=9.0Hz,1H), 7.12-7.06(m,4H), 6.90(dd,J=1.0,5.5Hz,1H), 6.75(s,1H), 5.75(bs,1H), 4.82(s,1H), 4.16-4.10(m,1H), 3.81(s,3H), 2.77-2.63(m,3H), 2.54-2.46(m,3H), 2.40-2.12(m,6H), 1.69(bs,4H);ESI MS, m / z=410[M+H] + .

[0111] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(5-methoxypyridin-3-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 9) [ka]

[0112] 1 H NMR (500 MHz, DMSO-d6) δ 8.15-8.11(m,2H), 7.56(d,J=9.2Hz,1H), 7.30-7.28(m,1H), 7.14-7.04 (m,4H), 5.76-5.71(m,1H), 4.92-4.89(m,1H), 4.17-4.09(m,1H), 3.78( s,3H), 2.83-2.70(m,2H), 2.69-2.60(m,1H), 2.50-2.48(m,4H), 2.47-2 .38(m,1H), 2.37-2.25(m,2H), 2.19-2.11(m,2H), 1.73-1.46(m,4H);ESI MS m / z 410[M+H] + .

[0113] 2-(2,3-Dihydro-1H-2-(2,3-dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)-N-methylacetamide (Example 10) [ka]

[0114] 1H NMR(500MHz,DMSO-d6)δ 8.15-8.05(m,1H), 7.36-7.33(m,2H), 7.15-7.06(m,4H), 5.61-4.11(m,3H), 3 .78-3.75(m,3H), 3.00-2.53(m,7H), 2.49-2.10(m,9H), 1.66-1.55(m,4H);ESI MS, m / z=424[M+H] +

[0115] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2S)-1-hydroxy-1-(5-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 11) [ka]

[0116] 1 H NMR(500MHz,DMSO-d6)δ 8.19-8.18(m,1H), 7.40-7.31(m,3H), 7.12-7.06(m,4H), 5.61(bs,1H), 4.83(s,1H), 4.30-4.20(m ESI MS, m / z=410[M+H] +

[0117] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-1-(4-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 12) [ka]

[0118] 1H NMR(500MHz,DMSO-d6)δ 8.29(s,1H), 7.39(d,J=9.0Hz,1H), 7.11-7.04(m,5H), 6.82(m,1H), 5.72(d,J=5.0Hz,1H), 4.84-4.83(m,1H), 4.33-4.29(m,1H) , 3.77(s,3H), 2.80-2.70(m,2H), 2.57-2.54(m,3H), 2.50-2.47(m,2H), 2.46-2.25(m,3H), 2.14-2.07(m,2H), 1.69(bs,4H);ESI MS, m / z=410[M+H] + .

[0119] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2R)-1-hydroxy-1-(pyridin-3-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 13) [ka]

[0120] 1 H NMR(500MHz,DMSO-d6)δ 8.52(d,J=2.1Hz,1H), 8.42(dd,J=1.6,4.7Hz,1H), 7.71-7.67(m,1H), 7.55(d,J =9.1Hz,1H), 7.30(dd,J=4.7,7.7Hz,1H), 7.16-7.05(m,4H), 5.73-5.69(m,1H), 4.91-4.88(m,1H), 4.17-4.10(m,1H), 2.82-2.61(m,3H), 2.57-2.49(m,5H), 2.4 6-2.39(m,1H), 2.36-2.24(m,2H), 2.14(d,J=7.5Hz,2H), 1.73-1.63(m,4H);ESI MS m / z 380[M+H] + .

[0121] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-1-(3-methoxypyridin-2-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamide (Example 14) [ka]

[0122] 1 H NMR(500MHz,DMSO-d6)δ 8.07(d,J=4.0Hz,1H), 7.42(d,J=8.0Hz,1H), 7.38-7.25(m,2H), 7.16-7.04(m,4H), 5.08(s,1H), 4.78(bs,1H), 4.47(b ESI MS, m / z=410[M+H] + .

[0123] N-((1S,2R)-3-(3-azabicyclo[3.1.0]hexan-3-yl)-1-hydroxy-1-(5-methoxypyridin-2-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 15) [ka]

[0124] 1 H NMR (500 MHz, DMSO-d6) δ 8.18(d,J=2.7Hz,1H), 7.39-7.29(m,3H), 7.14-7.05(m,4H), 5.53(d,J=5 .2Hz,1H), 4.77-4.73(m,1H), 4.22-4.14(m,1H), 3.79(s,3H), 3.05-2.97 (m,2H), 2.79-2.71(m,1H), 2.70-2.60(m,2H), 2.43-2.22(m,5H), 2.14-2 .05(m,2H), 1.39-1.28(m,2H), 0.59-0.54(m,1H), 0.32-0.26(m,1H);ESI MS m / z 421[M+H] + .

[0125] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-3-(3,3-dimethylpyrrolidin-1-yl)-1-hydroxy-1-(5-methoxypyridin-2-yl)propan-2-yl)acetamide (Example 16) [ka]

[0126] 1 H NMR(500MHz,DMSO-d6)δ 8.19(d,J=3.0Hz,1H), 7.39(d,J=8.5Hz,1H), 7.33(m,2H), 7.13-7.06(m,4H), 5.60(bs,1H), 4.83(s,1H), 4.22(bs,1H), 3 ESI MS, m / z=438[M+H] + .

[0127] N-((1S,2R)-1-(5-(tert-butoxy)pyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 17) [ka]

[0128] 1 H NMR(500MHz,DMSO-d6)δ 8.14(dd,J=1.0,2.4Hz,1H), 7.39-7.35(m,3H), 7.12-7.06(m,4H), 5.65(bs,1H), 4.85(s,1H), 4.28- 4.18(m,1H), 2.80-2.50(m,5H), 2.49-2.25(m,6H), 2.12-2.02(m,2H), 1.69(bs,4H), 1.29(s,9H);ESI MS, m / z=452[M+H] + .

[0129] N-((1R,2S)-1-(5-(tert-butoxy)pyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 18) [ka]

[0130] 1 H NMR(500MHz,DMSO-d6)δ 8.16-8.15(m,1H), 7.55-7.40(m,3H), 7.12-7.07(m,4H), 5.80(bs,1H), 4.83(s,1H) , 4.40(bs,1H), 3.10-2.50(m,7H), 2.49-2.55(m,6H), 1.75(bs,4H), 1.29(s,9H);ESI MS, m / z=452[M+H] + .

[0131] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-3-((R)-3-fluoropyrrolidin-1-yl)-1-hydroxy-1-(5-methoxypyridin-2-yl)propan-2-yl)acetamide (Example 19) [ka]

[0132] 1 H NMR(500MHz,DMSO-d6)δ 8.19(d,J=2.7Hz,1H), 7.43-7.36(m,2H), 7.35-7.31(m,1H), 7.13-7.03(m, 4H), 5.60(bs,1H), 5.27-5.10(m,1H), 4.84-4.81(m,1H), 4.29-4.18(m,1H) , 3.80(s,3H), 3.30-3.23(m,1H), 2.92-2.70(m,4H), 2.69-2.51(m,2H), 2.4 4-2.31(m,3H), 2.30-2.22(m,1H), 2.18-2.03(m,3H), 197-1.80(m,1H);ESI MS m / z 428[M+H]+ .

[0133] N-((1S,2R)-1-(6-chloropyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 20) [ka]

[0134] 1 H NMR(500MHz,DMSO-d6)δ 7.80(t,J=7.5Hz,1H), 7.45(d,J=7.5Hz,2H), 7.35(d,J=8.0Hz,1H), 7.12-7.08(m,4H), 5.84(bs,1H), 4.82(s,1 ESI MS, m / z=414[M+H] + .

[0135] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1S,2R)-1-hydroxy-3-(pyrrolidin-1-yl)-1-(5-(trifluoromethoxy)pyridin-2-yl)propan-2-yl)acetamide (Example 21) [ka]

[0136] 1H NMR(500MHz,DMSO-d6)δ 8.56(d,J=2.7,1H), 7.86-7.82(m,1H), 7.60(d,J=8.6Hz,1H), 7.45(d,J=9.2Hz,1H) , 7.12-7.04(m,4H), 5.87-5.83(m,1H), 4.95-4.90(m,1H), 4.35-4.27(m,1H), 2.84-2 .71(m,2H), 2.66-2.59(m,1H), 2.58-2.52(m,2H), 2.51-2.42(m,2H), 2.41-2.34(m, 1H)、2.33-2.27(m,1H)、2.26-2.17(m,1H)、2.14-2.04(m,2H)、1.73-1.62(m,4H);ESI MS m / z 464[M+H] + .

[0137] 2-(2,3-Dihydro-1H-inden-2-yl)-N-((1R,2S)-1-hydroxy-3-(pyrrolidin-1-yl)-1-(5-(trifluoromethoxy)pyridin-2-yl)propan-2-yl)acetamide (Example 22) [ka]

[0138] 1 H NMR(500MHz,DMSO-d6)δ 8.56(d,J=2.7,1H), 7.87-7.82(m,1H), 7.60(d,J=8.6Hz,1H), 7.46(d,J=9.3Hz ,1H), 7.12-7.05(m,4H), 5.87-5.83(m,1H), 4.95-4.91(m,1H), 4.35-4.28(m,1H) ), 2.84-2.71(m,2H), 2.66-2.52(m,3H), 2.51-2.43(m,2H), 2.42-2.34(m,1H), 2.33-2.27(m,1H), 2.26-2.19(m,1H), 2.15-2.03(m,2H), 1.72-1.64(m,4H);ESI MS m / z 464[M+H] + .

[0139] N-((1S,2R)-1-(5-(tert-butoxy)pyridin-2-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(5-fluoro-2,3-dihydro-1H-inden-2-yl)acetamide (Example 24) [ka]

[0140] 1 H NMR(500MHz,DMSO-d6)δ 8.14(d,J=2.0Hz,1H), 7.45-7.30(m,3H), 7.15-6.85(m,3H), 5.65(bs,1H), 4.84(s,1H), 4.29(b ESI MS, m / z=470[M+H] + .

[0141] N-((1R,2R)-1-(2-chloropyridin-4-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 27) [ka]

[0142] 1 H NMR(400MHz,CDCl3)δ 8.38(d,J=5Hz,1H), 7.42(s,1H), 7.13-7.22(m,5H), 5.91(d,J=7.6Hz,1H), 5.12(d,J=2Hz,1H), 4.33 (m,1H), 2.8-3.2(m,9H), 2.51(dd,J=6,16Hz,1H), 2.31(m,2H), 2.18(dd,J=8,14Hz,1H), 1.84(m,5H). ESI MS, m / z=414.1[M+H] + .

[0143] GCS inhibition GCS inhibitors are known. Some GCS inhibitors, such as eliglustat and miglustat, have sufficient activity to inhibit GCS activity and have therefore been proposed as suitable for the treatment of diseases associated with glycolipid accumulation. However, these compounds and / or their pharmacological profiles are not entirely satisfactory. For example, miglustat can cross the blood-brain barrier (BBB), but its IC 50

[0003] Eliglustat does not achieve levels exceeding 0.05% and many of its effects are either off-target or attributable to its potential activity as a chemical chaperone for beta-glucocerebrosidase. Although eliglustat is more potent than miglustat in inhibiting GCS, it cannot cross the BBB. Therefore, diseases that require therapeutic agents to cross the BBB cannot be treated. Consequently, there is a continuing need to provide novel compounds that effectively and selectively inhibit GCS and, in some embodiments, can cross the BBB. Selected compounds of structural formula (I) exhibit these beneficial properties. At the same time, there remains a need to treat non-CNS diseases, such as type 1 Gaucher disease and Fabry disease, with potent GCS inhibitors that do not efficiently cross the BBB. Selected compounds of structural formula (I) exhibit this property.

[0144] To demonstrate the ability of the GCS inhibitors of the present invention to reduce glycolipid accumulation in lysosomes and assess their ability to cross the BBB, compounds of the present invention were prepared and tested in in vitro and in vivo assays. In vitro assays measured the inhibition of glucosylceramide formation in cell homogenates and the reduction in glucosylceramide content in Madin-Darby canine kidney (MDCK) cells 24 hours after inhibitor addition. In vivo assays measured the reduction in brain and liver glucosylceramide and glucosylsphingosine levels in mice on a D409V background treated in parallel with conduritol B epoxide (CBE), an irreversible inhibitor of GBA (beta-glucocerebrosidase). Compounds of structural formula (I) are also more potent GCS inhibitors in cells and exhibit improved metabolic stability compared to prior art GCS inhibitors.

[0145] Compounds of structural formula (I) have been found to inhibit GCS in vitro at low nanomolar concentrations and to reduce glycosphingolipid levels in vivo with varying degrees of selectivity for brain and liver.

[0146] Assay GCS inhibition in MDCK cell homogenates (lysate IC50, Table 1) The IC50 values of the novel compounds of the present invention were determined as reported in Shayman JA, Abe A. 2000. Glucosylceramide synthase: assay and properties. Methods Enzymol. 311:42-49.

[0147] GCS inhibition in MDCKII cells (MDCKII IC50, Table 1) MDCKII cells were routinely maintained in a medium consisting of Opti-MEM / F12 (1:1), 5% FBS, 100 U / mL penicillin, 100 μg / mL streptomycin, and 200 mM L-glutamine. MDCKII cells were freshly thawed from frozen ampoules every 2 months.

[0148] Stock solutions of water-insoluble glycosphingolipid inhibitors (100 mM) were prepared by dissolving each inhibitor in 100% ethanol as previously described (3). The inhibitor-ethanol solution was then diluted 50-fold into 2 mM delipidated bovine serum albumin-phosphate-buffered saline to produce water-soluble glycosphingolipid inhibitor-bovine serum albumin conjugates. The inhibitor-bovine serum albumin conjugates were sterile filtered and stored at -20°C. Before use, a portion of the inhibitor-bovine serum albumin conjugate was further diluted with Opti-F12 to produce treatment solutions. Equal volumes of bovine serum albumin and ethanol were added to control cultures. Cells (5 × 10 cells) were plated in a 10 cm culture dish containing 10 ml of Opti-F12 with 5% FBS. 5 After 24 hours, the medium was replaced with fresh serum-free Opti-F12 medium, and the cells were exposed to candidate GCS inhibitors at concentrations of 0, 1, 3, 10, 30, 100, and 300 nM for 24 hours.

[0149] Cellular lipid analysis After inhibitor treatment, total cellular lipids from wild-type MDCKII cells were extracted as previously described in detail (10). Briefly, cells were washed with ice-cold phosphate-buffered saline, fixed with methanol, and collected with a rubber scraper. Chloroform was then added to obtain a theoretical chloroform:methanol:water ratio of 1:2:0.8 (v / v / v) to form a single phase. Cell debris and proteins were removed by centrifugation at 2200 × g for 30 min. The supernatant was partitioned by adding chloroform and 0.9% NaCl. The lower organic phase, containing neutral glycosphingolipids, was washed with methanol and 0.9% NaCl and subjected to base and acid hydrolysis (10). A portion of the purified glycosphingolipids, normalized to 100 nmol of total phospholipids, was analyzed by high-performance thin-layer chromatography. Thin-layer chromatography separations were performed twice. Plates pretreated with 1% sodium borate were first developed with a solvent system consisting of chloroform / methanol (98 / 2, v / v). After air drying, plates were developed in a solvent system containing chloroform / methanol / water (70 / 30 / 4, v / v / v). Glucosylceramide levels were detected by charring with 8% copper sulfate in 8% phosphoric acid and quantified by densitometric scanning using ImageJ, NIH Image. Image data were analyzed, and the IC values for each inhibitor were calculated using GraphPad Prism (version 5.03). 50 was calculated, and the results are summarized in Table 1.

[0150] Decreased glucosylceramide and glucosylsphingosine in brain and liver in CBE mice The reduction of brain glucosylceramide and glucosylsphingosine content by inhibitors of GCS was determined using mice carrying a Gba1 mutant allele, designated D409V / null. These are knock-in mice with a Gba1 point mutation encoding valine (V) at 409 instead of wild-type (WT) aspartic acid (D) on one missense allele and a null heterozygous allele [D409V / null (9V / null)]. D409V / null and WT littermates were of mixed sex and mixed but matched C57BL / 129Sv / FVB genetic backgrounds. (Xu YH, Sun Y, Barnes S, Grabowski GA (2010) Comparative therapeutic effects of velaglucerase alfa and imiglucerase in a Gaucher disease mouse model. PLoS One 5:e10750.) Starting on postnatal day 15, all groups of mice were treated intraperitoneally with conduritol B epoxide (CBE) (25 mg / kg / day), an irreversible inhibitor of beta-glucocerebrosidase. Mice were treated in parallel with GCS inhibitors or vehicle control for 14 days. Mice were sacrificed 2 hours after the last injection, and tissues including brain, liver, spleen, kidney, and lung were analyzed for glucosylceramide and glucosylsphingosine by mass spectrometry.

[0151] Mouse tissue lipid analysis Lipid extraction from liver, kidney, and brain was performed as previously described (7). Briefly, frozen liver (approximately 0.5 g), two kidneys (approximately 0.3 g), and whole brain (approximately 0.4 g) were individually homogenized using a Tri-R homogenizer in 0.2 g tissue / 1 mL of sucrose buffer (250 mM sucrose, pH 7.4, 10 mM HEPES, and 1 mM EDTA). 0.8 mL of each homogenate was mixed with 2 mL of methanol and 1 mL of chloroform, sonicated for 1 minute, and incubated at room temperature for 1 hour. Tissue debris was removed by centrifugation at 2,400 × gravity for 30 minutes. The pellet was re-extracted by mixing with 1 mL of methanol, 0.5 mL of chloroform, and 0.4 mL of 0.9% NaCl (chloroform / methanol / 0.9% NaCl, 1:2:0.8), incubated at room temperature for 1 hour, and centrifuged at 2,400 × gravity for an additional 30 minutes. The two extracts were combined and mixed with 4.5 mL of chloroform and 1.2 mL of 0.9% NaCl (chloroform / methanol / 0.9% NaCl, 2:1:0.8). After centrifugation at 800 × gravity for 5 minutes, the lower layer was washed with 3 mL of methanol and 2.4 mL of 0.9% NaCl. A second wash was performed with 3 mL of methanol, 2 mL of water, and 0.4 mL of 0.9% NaCl, followed by centrifugation at 800 × gravity for 5 minutes. The resulting lower phase was collected and dried under a stream of N2 gas.

[0152] Neutral glycosphingolipids from mouse liver, kidney, and brain were analyzed after alkaline methanolysis. Kidney lipids were incubated with 1 ml of 0.21 N NaOH in 2 ml of chloroform and methanol at room temperature for 2 h (kidney) or 7.5 h (liver and brain). For high-performance thin-layer chromatography analysis, lipid extracts were normalized to 0.5 μmol of total phospholipid phosphate (liver and kidney) or 2 μmol of total phospholipid phosphate (brain). After alkaline methanolysis, brain lipids were passed through a silica gel column (7). Borate-impregnated thin-layer chromatography plates were developed with two solvent systems: plates were first developed in chloroform / methanol (98:2, v / v). The kidney and liver lipid-loaded plates were then developed in chloroform / methanol / water (64:24:4, v / v / v), and brain lipids were further separated in chloroform / methanol / water (60:30:6, v / v / v). GlcCer levels were quantified by comparison with known standards.

[0153] Assay Results The activity of compounds of structural formula (I) in inhibiting GlcCer production in vitro is summarized in Table 1. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5]

[0154] The activity of selected compounds of structural formula (I) in inhibiting GlcCer production in vivo is summarized in Table 2. [Table 4]

[0155] The pharmacokinetics of selected prior art compounds of formula (I) and structurally related compounds lacking an aromatic nitrogen atom are summarized in Table 3. This demonstrates that incorporating nitrogen into the aromatic ring can reduce the brain / plasma ratio in mice, indicating that selected compounds of the present invention may be advantageous in treating peripheral glycolipid storage diseases such as type 1 Gaucher disease and Fabry disease. [Table 5]

[0156] Methods and Compositions The present invention provides GCS inhibitors, exemplified by compounds of structural formula (I), for the treatment of various diseases and conditions in which inhibition of the GCS has a beneficial effect. In one embodiment, the present invention relates to a method of treating an individual suffering from a disease or condition in which inhibition of the GCS would be beneficial, comprising administering to an individual in need thereof a therapeutically effective amount of a compound of structural formula (I).

[0157] Therefore, compounds of structural formula (I) can be used to treat a variety of diseases and conditions that benefit from GCS inhibition, including, but not limited to, Tay-Sachs disease, types I, II, and III Gaucher disease, Sandhoff disease, and Fabry disease; Parkinson's disease (JR Mazzulli et al., Cell 146:37-52, July 8, 2011); type 2 diabetes; renal hypertrophy or hyperplasia associated with diabetic nephropathy; elevated plasma TNF-α; elevated blood glucose; elevated glycosylated hemoglobin; lupus; and glomerular diseases selected from the group consisting of mesangial proliferative glomerulonephritis, collapsing glomerulopathy, proliferative lupus nephritis, crescentic glomerulonephritis, and membranous nephropathy, or viral diseases.

[0158] Compounds of structural formula (I) are useful for treating disorders involving cell proliferation and division, including cancer, collagen vascular disease, atherosclerosis, and renal hypertrophy in diabetic individuals (U.S. Pat. Nos. 6,916,802 and 5,849,326, each of which is incorporated herein by reference); for inhibiting the proliferation of arterial endothelial cells (U.S. Pat. Nos. 6,916,802 and 5,849,326, each of which is incorporated herein by reference); for treating patients suffering from infectious diseases (M. Svensson et al., Infect. And Immun., 62:4404-4410 (1994)); for preventing the host, i.e., patient, from producing antibodies against tumors (J. Inokuchi et al., Cancer Lett., 38:23-30 (1987)); and for treating tumors (S. Hakomori Cancer Cells 3:461-470 (1991)); J. Inokuchi et al., Cancer Res., 50L6731-6737(1990)); and (M. Ziche et al., Lab Invest., 67:711-715(1992)). The compounds of structural formula (I) can also be used to treat polycystic kidney disease, including both autosomal dominant and recessive forms (TA Natoli et al., Nat. Med. 16:788-792(2010)).

[0159] The methods of the present invention can be achieved by administering a compound of structural formula (I) as a pure compound or as a pharmaceutical composition. Administration of the pharmaceutical composition or pure compound of structural formula (I) can occur during or after the onset of the disease or condition of interest. Typically, the pharmaceutical composition is sterile and does not contain toxic, carcinogenic, or mutagenic compounds that would cause adverse reactions when administered. Kits are further provided that contain, packaged separately or together, a compound of structural formula (I) and, optionally, a second therapeutic agent useful in treating diseases and conditions benefiting from inhibition of GCS, as well as a package insert with instructions for using these active agents.

[0160] In many embodiments, a compound of structural formula (I) is administered in conjunction with a second therapeutic agent useful in the treatment of a disease or condition benefiting from inhibition of GCS. The second therapeutic agent is different from the compound of structural formula (I). The compound of structural formula (I) and the second therapeutic agent may be administered simultaneously or sequentially to achieve the desired effect. Furthermore, the compound of structural formula (I) and the second therapeutic agent may be administered from a single composition or two separate compositions.

[0161] The second therapeutic agent is administered in an amount that produces its desired therapeutic effect. Effective dosage ranges for each second therapeutic agent are known in the art, and the second therapeutic agent is administered to an individual in need thereof within such established ranges.

[0162] The compound of structural formula (I) and the second therapeutic agent can be administered together in a single unit dose or separately in multiple unit doses, with the compound of structural formula (I) administered before the second therapeutic agent, or vice versa. One or more doses of the compound of structural formula (I) and / or one or more doses of the second therapeutic agent can be administered. Thus, the compound of structural formula (I) can be used in conjunction with one or more second therapeutic agents, such as, but not limited to, enzyme replacement therapy, gene therapy, and isofagamine.

[0163] In methods for treating type 2 diabetes, the second therapeutic agent can be one or more of the following concomitant medications: insulin (e.g., NOVOLIN®, NOVOLOG®, VELOSULIN®); sulfonylureas (e.g., DIABINESE®, GLUCOTROL®, GLUCOTROL XL®, DIABETA®, AMARYL®, ORINASE®, TOLINASE®, MICRONASE®, and GLYNASE®); metformin; [alpha]-glucosidase inhibitors (e.g., GLYSET®); thiazolidinediones (e.g., ACTOS® and AVANDIA®), nateglinide (STARLIX®); repaglinide (PRANDIN®), and AVANDAMET® (AVANDIA® and metformin).

[0164] In methods for treating Parkinson's disease, the second therapeutic agent can be one or more of carbidopa / levodopa therapy; other medications including dopamine agonists (apomorphine hydrochloride, bromocriptine, rotigotine, pramipexole, ropinirole, pergolide), anticholinergics (benzotropine mesylate, trihexyphenidyl hydrochloride, procyclidine), MAO-B inhibitors (selegiline, rasagiline), COMT inhibitors (entacapone, tulcapone), and non-prescription, over-the-counter medications (amantadine, rivastigmine tartrate, creatine, coenzyme Q10).

[0165] Diseases and conditions that can be treated according to the present invention include, for example, Gaucher disease, Fabry disease, Tay-Sachs disease, polycystic kidney disease, and diabetes. In particular, the compound of structural formula (I) can cross the BBB and therefore can treat types II and III of Gaucher disease. Prior art GCS inhibitors either cannot cross the BBB or have low efficacy and selectivity, making them unable to treat various diseases associated with glycolipid accumulation.

[0166] In the methods of the present invention, a therapeutically effective amount of one or more compounds of structural formula (I), typically formulated according to pharmaceutical practice, is administered to a human in need thereof. Whether such treatment is indicated will depend on the individual case and is subject to a medical evaluation (diagnosis) that takes into account the signs, symptoms, and / or dysfunctions present, the risk of developing the particular signs, symptoms, and / or dysfunctions, and other factors.

[0167] Compounds of structural formula (I) can be administered by any suitable route, for example, oral, buccal, inhalation, sublingual, rectal, vaginal, intracisternal or intrathecal via lumbar puncture, urethral, nasal, percutaneous (i.e., transdermal), or parenteral (including intravenous, intramuscular, subcutaneous, intracoronary, intradermal, intramammary, intraperitoneal, intraarticular, intrathecal, retrobulbar, intrapulmonary injection, and / or surgical implantation at a particular site) administration. Parenteral administration can be accomplished using a needle and syringe or using high pressure techniques.

[0168] Pharmaceutical compositions include those in which a compound of structural formula (I) is administered in an effective amount to achieve its intended purpose. The exact formulation, route of administration, and dosage will be determined by the individual physician in light of the diagnosed condition or disease. Dosage amount and interval can be individually adjusted to provide a sufficient level of the compound of structural formula (I) to maintain therapeutic effect.

[0169] The toxicity and therapeutic efficacy of compounds of structural formula (I) can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the maximum tolerated dose (MTD) of the compound, defined as the highest dose that does not cause toxicity in animals. The dose ratio between the maximum tolerated dose and the therapeutic effect (e.g., inhibition of tumor growth) is the therapeutic index. Dosages can vary within this range depending on the dosage form and route of administration used. Determining a therapeutically effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0170] The therapeutically effective amount of the compound of formula (I) required for use in therapy varies depending on the nature of the condition being treated, the length of time for which activity is desired, and the age and condition of the patient, and is ultimately determined by the attending physician. The amount and interval of administration can be individually adjusted to provide a plasma level of the compound of formula (I) sufficient to maintain the desired therapeutic effect. The desired dose can conveniently be administered in a single dose or as multiple doses administered at appropriate intervals, for example, as divided doses once, twice, three times, four or more times per day. Often, multiple doses are desired or required. For example, a compound of structural formula (I) can be administered at a frequency of four doses delivered as one dose per day spaced four days apart (q4d×4); four doses delivered as one dose per day spaced three days apart (q3d×4); one dose per day spaced five days apart (qd×5); one dose per week for three weeks (qwk3); five daily doses with two days of rest and five more daily doses (5 / 2 / 5); or any dosing regimen determined to be appropriate for the circumstances.

[0171] The compounds of structural formula (I) used in the methods of the invention can be administered in amounts of about 0.005 to about 500 milligrams per dose, about 0.05 to about 250 milligrams per dose, or about 0.5 to about 100 milligrams per dose. For example, the compounds of structural formula (I) can be administered in amounts of about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 milligrams per dose, including all doses from 0.005 to 500 milligrams.

[0172] The dosage of a composition containing a GCS inhibitor of structural formula (I), or a composition containing the same, can be about 1 ng / kg to about 200 mg / kg, about 1 μg / kg to about 100 mg / kg, or about 1 mg / kg to about 50 mg / kg. The dosage of the composition can be any dosage, including, but not limited to, about 1 μg / kg. The dosage of the composition can be about 1 μg / kg, 10 μg / kg, 25 μg / kg, 50 μg / kg, 75 μg / kg, 100 μg / kg, 125 μg / kg, 150 μg / kg, 175 μg / kg, 200 μg / kg, 225 μg / kg, 250 μg / kg, 275 μg / kg, 300 μg / kg, 325 μg / kg, 35 μg / kg, 400 μg / kg, 450 μg / kg, 460 μg / kg, 475 μg / kg, 480 μg / kg, 490 μg / kg, 500 μg / kg, 510 μg / kg, 520 μg / kg, 530 μg / kg, 540 μg / kg, 550 μg / kg, 560 μg / kg, 570 μg / kg, 580 μg / kg, 590 μg / kg, 600 μg / kg, 610 μg / kg, 620 μg / kg, 630 μg / kg, 640 μg / kg, 650 μg / kg, 660 μg / kg, 670 μg / kg, 680 μg / kg, 690 μg / kg, 700 μg / kg, 710 μg / kg, 720 μg / kg, 730 μ 0μg / kg, 375μg / kg, 400μg / kg, 425μg / kg, 450μg / kg, 475μg / kg, 500μg / kg, 525μg / kg, 550μg / kg, 575μg / kg, 600μg / kg, 625μg / kg, 650μg / kg, 675μg / kg, 700μg / kg, 725μg / kg , 750 μg / kg, 775 μg / kg, 800 μg / kg, 825 μg / kg, 850 μg / kg, 875 μg / kg, 900 μg / kg, 925 μg / kg, 950 μg / kg, 975 μg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 175 mg / kg, or 200 mg / kg. While the above dosages are exemplary of the average case, there may be individual instances in which higher or lower dosages are suitable, and such instances are within the scope of this invention. In practice, the physician will determine the actual dosing regimen that will be most suitable for an individual patient, which may vary with the age, weight, and response of the particular patient.

[0173] The compounds of the invention are typically administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. Pharmaceutical compositions for use in accordance with the invention are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and adjuvants that facilitate processing of the compounds of structural formula (I).

[0174] These pharmaceutical compositions can be prepared, for example, by conventional mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating, entrapping, or lyophilizing processes. Suitable formulations depend on the route of administration chosen. When a therapeutically effective amount of a compound of structural formula (I) is administered orally, the composition is typically in the form of a tablet, capsule, powder, solution, or elixir. When administered in tablet form, the composition may further contain a solid carrier such as gelatin or an adjuvant. Tablets, capsules, and powders contain about 0.01% to about 95%, preferably about 1% to about 50%, of a compound of structural formula (I). When administered in liquid form, a liquid carrier such as water, petroleum, or an oil of animal or vegetable origin can be added. Liquid forms of the composition may further contain physiological saline solution, dextrose or other sugar solution, or glycol. When administered in liquid form, the composition contains about 0.1% to about 90%, preferably about 1% to about 50%, by weight of a compound of structural formula (I).

[0175] When a therapeutically effective amount of a compound of structural formula (I) is administered by intravenous, cutaneous, or subcutaneous injection, the composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such a parenterally acceptable solution is within the skill of one in the art, taking into due consideration pH, isotonicity, stability, etc. Preferred compositions for intravenous, cutaneous, or subcutaneous injection typically contain an isotonic vehicle.

[0176] The compound of formula (I) can be easily combined with pharmaceutically acceptable carriers well known in the art. Such carriers allow the active agent to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral ingestion by the patient to be treated. Oral medicaments can be obtained by adding the compound of formula (I) to a solid excipient, optionally grinding the resulting mixture, and, if necessary, adding suitable adjuvants, followed by processing the granular mixture to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. If necessary, disintegrants can be added.

[0177] The compound of structural formula (I) can be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion. The injection preparation can be presented in unit dosage form, for example, in ampoules or multi-dose containers, with added preservatives. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and can contain formulatory agents such as suspending, stabilizing, and / or dispersing agents.

[0178] Pharmaceutical compositions for parenteral administration include aqueous solutions of the active agent in water-soluble form. Furthermore, suspensions of the compounds of structural formula (I) can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound, allowing for the preparation of highly concentrated solutions. Alternatively, the compositions of the present invention can be in powder form for constitution with a suitable vehicle, for example, sterile pyrogen-free water, before use.

[0179] The compounds of structural formula (I) can also be formulated into rectal compositions, such as suppositories or retention enemas, containing conventional suppository bases.In addition to the above-mentioned formulations, the compounds of structural formula (I) can also be formulated as depot preparations.Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection.Thus, for example, the compounds of structural formula (I) can be formulated with suitable polymers or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins.

[0180] Specifically, the compound of structural formula (I) can be administered orally, bucally, or sublingually in the form of tablets containing excipients such as starch or lactose, or in the form of capsules or suppositories alone or mixed with excipients, or in the form of elixirs or suspensions containing flavorings or colorings. Such liquid preparations can be prepared with pharmaceutically acceptable additives such as suspending agents. The compound of structural formula (I) can also be injected parenterally, for example, intravenously, intramuscularly, subcutaneously, or intracoronarily. For parenteral administration, the GCS inhibitor is best used in the form of a sterile aqueous solution, which may contain other substances, such as salts or simple sugars such as mannitol or glucose, to make the solution isotonic with blood.

[0181] In additional embodiments, the present invention includes kits containing one or more compounds or compositions packaged in a manner that facilitates their use to practice the methods of the invention. In one simple embodiment, the kit includes a compound or composition described herein (e.g., a composition comprising a compound of structural formula (I) and an optional second therapeutic agent) useful for practicing the methods, packaged in a container such as a sealed bottle or container, with a label affixed to the container or included in the kit that describes the use of the compound or composition to practice the methods of the invention. Preferably, the compound or composition is packaged in a unit dosage form. The kit can further include a device suitable for administering the composition according to the intended route of administration.

[0182] Prior art GCS inhibitors have properties that prevent their development as therapeutic agents. In accordance with an important feature of the present invention, compounds of structural formula (I) have been synthesized and evaluated as inhibitors of GCS, particularly for their ability to cross the BBB. The GCS inhibitors of the present invention are characterized by low nanomolar GCS inhibition, high specificity, and the absence of β-glucocerebrosidase binding.

[0183] References 1. NW Barton et al., N Engl J Med324, 1464-1470, (1991). 2.NS Radin, Glycoconj J13, 153-157, (1996). 3. JAShayman et al., Methods Enzymol311, 373-387, (2000). 4.NJWeinreb et al.,Am J Hematol80,223-229,(2005). 5. JAShayman, Drugs of the Future35, 613-621, (2010). 6. E. Lukina et al., Blood116(20):4095-8, (2010). 7. A. Abe et al., J Clin Invest105, 1563-1571, (2000). 8. Y. Liu et al., The Journal of clinical investigation 103, 497-505, (1999). 9. JAShayman et al., Methods Enzymol311, 42-49, (2000). 10. L. Shu et al., J Biol Chem278, 31419-31425, (2003).

Claims

1. A compound having the structure: 【Chemical 1】 During the ceremony, 【Chemistry 2】 but, 【Chemistry 3】 and Each X is independently CH or CR １ and A is CH ２ or O, R １ is H, halogen, or OR ５ and R ２ But H, C １ -C ５ Alkyl, or C ３ -C ６ is cycloalkyl, R ３ is H or CH ３ and R ４ is H or F, R ５ optionally substituted with one or more F; １ -C ４ Alkyl or C ３ -C ６ is cycloalkyl, R ６ and R ７ are independently H or C １ -C ３ alkyl or R ６ and R ７ together with the nitrogen atom to which they are attached, optionally F or CH ３ C substituted with ４ -C ８ Compounds forming a heterocycloalkyl or heterobicycloalkyl ring, or a pharmaceutically acceptable salt thereof.

2. R １ -H, -OC(CH ３ ) ３ , -OCH ３ , -OCF ３ , -OCH ２ CF ３ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

3. R ２ is H or CH ３ 2. The compound of claim 1, wherein:

4. NR ６ R ７ but, 【Chemistry 4】 2. The compound of claim 1, wherein: 【Request 5】 【Chemical 5】 but, 【Chemistry 6】 2. The compound of claim 1, wherein: 【Request 6】 【Chemical 7-1】 【Chemistry 7-2】 or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or vehicle.

8. (a) a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof; (b) a second therapeutic agent useful in the treatment of a disease or condition benefiting from inhibition of GCS, and (c) an optional excipient and / or pharmaceutically acceptable carrier.

9. 9. The composition of claim 8, wherein the second therapeutic agent comprises an agent useful in the treatment of Gaucher disease, Fabry disease, type II diabetes, Sandhoff disease, Tay-Sachs disease, or Parkinson's disease.

10. A medicament for treating a disease or condition benefiting from inhibition of GCS, comprising a therapeutically effective amount of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof.

11. The method of claim 10, further comprising administering a therapeutically effective amount of a second therapeutic agent useful in said treatment of said disease or condition.

12. The drug of claim 10 or 11, wherein the disease or condition is Gaucher disease, Fabry disease, Sandhoff disease, Tay-Sachs disease, Parkinson's disease, multiple myeloma, ADPCD, type 2 diabetes, hypertrophy or hyperplasia associated with diabetic neuropathy, elevated plasma TNF-α levels, elevated blood glucose levels, elevated glycated hemoglobin levels, glomerular disease, or lupus.

13. 13. The method of claim 12, wherein the Gaucher disease is type I, type II, or type III Gaucher disease.

14. The method of claim 12, wherein the glomerular disease is selected from the group consisting of mesangial proliferative glomerulonephritis, collapsing glomerulopathy, proliferative lupus nephritis, crescentic glomerulonephritis, and membranous nephropathy.

15. The drug described in claim 10 or 11, wherein the disease or condition is a disorder involving cell proliferation, a disorder involving cell division, collagen vascular disease, atherosclerosis, renal hypertrophy in diabetic individuals, proliferation of arterial endothelial cells, infection, tumor, and polycystic kidney disease.

16. 16. The method of claim 15, wherein the disease or condition is cancer or an autosomal dominant or recessive form of polycystic kidney disease.

17. 12. The method of claim 11, wherein the second therapeutic agent is one or more of enzyme replacement therapy, gene therapy, and isotagamine.

18. 12. The method of claim 11, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof and the second therapeutic agent are administered simultaneously.

19. 12. The method of claim 11, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof and the second therapeutic agent are administered separately.

20. 12. The method of claim 11, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof is administered before the second therapeutic agent.

21. The method of claim 11, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof is administered after the second therapeutic agent.

22. 12. The method of claim 11, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof and the second therapeutic agent are administered from a single composition.

23. 12. The method of claim 11, wherein the compound of claim 1 or a pharmaceutically acceptable salt thereof and the second therapeutic agent are administered from separate compositions.

24. A drug for inhibiting glucosylceramide synthase or reducing glycosphinolipid levels, comprising a therapeutically effective amount of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof.

Images