Glucosylceramide synthase inhibitor and therapeutic method using the same

Abstract

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

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Application No. 62 / 924,959, filed October 23, 2019, which is hereby incorporated by reference in its entirety.

[0002] Government Funding This invention was made with government support under Grant No. NS092981 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0003] The present invention relates to glucosylceramide synthase (GCS) inhibitors and methods of treatment for treating conditions and diseases in which inhibition of GCS provides a benefit.

Background Art

[0004] Lysosomal storage diseases (LSDs), such as Gaucher disease and Fabry disease, occur when glycolipids accumulate in lysosomes due to defects in catabolism. There are two general strategies for treating lysosomal storage diseases. The first strategy involves replacement or restoration of defective or absent catabolic enzymes (e.g., infusion of recombinant enzymes, chaperone therapy, bone marrow transplantation, or gene therapy)(1). Enzyme replacement therapy is clinically approved for lysosomal storage diseases with peripheral symptoms, but is limited by the inability of infused recombinant enzymes to distribute into the CNS and the frequent expression of autoantibodies against the protein in patients with null mutations.

[0005] The second strategy involves synthetic inhibitor therapies focused on the identification of small molecule inhibitors of GCS (2). Various classes of GCS inhibitors have been described, including iminosugars and analogs of 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 synthase. The limited specificity is associated with high levels of unwanted effects due to secondary sites of action not related to glycolipid synthesis inhibition. These effects include, most prominently, diarrhea, weight loss, and tremors, and in the United States, the approved use of NBDNJ is limited (4). One advantage of NBDNJ over previously reported PDMP-based homologs is its ability to distribute to the CNS. However, recent studies have questioned NBDNJ's ability to lower CNS glycolipid levels (K.M. Ashe et al., Plos One 6:e21758 (2011)).

[0006] 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, 6,916,802, 7,253,185, 7,196,205, 7,615,573, and 10,202,340. Further GCS inhibitors and therapies are disclosed in WO2008 / 150486, WO2009 / 117150, WO2010 / 014554, and WO2012 / 129084.

[0007] Compounds structurally related to PDMP are 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). In a phase 2 clinical trial using this drug for type 1 Gaucher disease, efficacy equal to or greater than that of recombinant β-glucocerebrosidase was shown, as evident from the reversal of spleen and liver enlargement, correction of anemia, and improvement of thrombocytopenia and bone density (6). A phase 3 trial using eliglustat tartrate has been published, and eliglustat tartrate is approved worldwide for type 1 Gaucher disease (7).

[0008] GSC inhibition is also expected to treat other six lysosomal storage diseases involving the CNS, including early and late-onset Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis, and type 2 and 3 Gaucher diseases. For example, an experimental model of genetic epistasis showed a significant improvement in survival in a mouse model of Sandhoff disease that also lacked GM2 synthase (8). However, drug distribution studies have shown that eliglustat tartrate is not transported across the blood-brain barrier (BBB) (5). It is considered that the reason for the insufficient brain distribution of eliglustat tartrate may be that the drug is a substrate of the p-glycoprotein (MDR1) transporter, which may result in drug efflux.

[0009] Compounds that inhibit GCS have the potential to treat conditions associated with the accumulation of glycolipids. However, current GCS inhibitors are limited due to insufficient penetration into the central nervous system and / or low activity. An important advance in the art is the discovery of GCS inhibitors, particularly those that can cross the BBB, which are useful in the treatment of diseases such as type I, II, or III Gaucher disease, Fabry disease, Tay-Sachs disease, Sandhoff disease, diabetes, lupus, and other diseases and conditions associated with the accumulation of glycolipids in lysosomes where GCS inhibition provides a benefit. Accordingly, there remains a need in the art for effective compounds, compositions, and methods useful in the treatment of such diseases, either alone or in combination with other therapies used to treat these diseases and conditions. The present invention is directed to meeting this need.

Summary of the Invention

[0010] The present invention is directed to inhibitors of GCS, methods of preparing GCS inhibitors, compositions containing the inhibitors, and methods of using the inhibitors in therapeutic treatment of conditions and diseases where inhibition of GCS provides a benefit. The compounds of the present invention are potent inhibitors of GCS and, in some embodiments, can cross the BBB.

[0011] More specifically, the present invention is a compound having the following structural formula (I),

Chemical

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

[0014] Also included in the present invention are methods of treating a subject having renal hypertrophy, hyperplasia associated with diabetic neuropathy, or autosomal dominant polycystic kidney disease (ADPKD). The methods comprise administering to the subject a therapeutically effective amount of a compound of 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 with the compounds of structural formula (I) include, but are not limited to, multiple myeloma (due to depletion of glucosylsphingosine) and inhibition of viral and bacterial diseases, including, for the latter, depletion of the receptor for Shiga toxin, i.e., globotriaosylceramide, which can be depleted from vascular endothelial cells, including hemolytic uremic syndrome. The method includes administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0016] Also included in the present invention is a method for reducing blood glucose level in a subject in need thereof. The method includes 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 in a subject in need thereof. The method includes 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 the concentration of sphingolipids in a subject in need thereof. The method includes 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, focal segmental glomerulosclerosis, proliferative lupus nephritis, crescentic glomerulonephritis, and membranous nephropathy in a subject, which includes 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 for treating lupus in a subject, which includes administering to the subject a therapeutically effective amount of a compound of structural formula (I).

[0021] In yet another embodiment, in the treatment of the above-disclosed diseases, the compound 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 invention provides a composition comprising (a) a GCS inhibitor of structural formula (I) and (b) an excipient and / or a pharmaceutically acceptable carrier useful in the treatment of a disease or condition in which inhibition of GCS provides a benefit.

[0023] Another embodiment of the invention is the use of a composition comprising a compound of structural formula (I) and a second therapeutic agent in a method of treating an individual for a disease or condition in which inhibition of GCS provides a benefit.

[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, such as Gaucher disease or Fabry disease.

[0025] Yet another embodiment of the invention provides a kit for human pharmaceutical use comprising (a) a container and (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 the treatment of a disease or condition of interest, and (c) a package insert containing instructions for the use of the composition or compositions to be administered simultaneously or sequentially in the treatment of the disease or condition.

[0026] The GCS inhibitor of structural formula (I) and the second therapeutic agent can be administered together as a single unit dose or separately as multiple unit doses, and the GCS inhibitor of structural formula (I) can be administered before or vice versa to the second therapeutic agent. It is contemplated that one or more doses of the GCS inhibitor of structural formula (I) and / or one or more doses 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 related embodiments, the GCS inhibitor of structural formula (I) and the second therapeutic agent are administered from a single composition or from separate compositions. In further embodiments, the GCS inhibitor of structural formula (I) and the second therapeutic agent are administered sequentially. The GCS inhibitor of structural formula (I) used in the methods of the invention can 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 invention inhibit GCS and are useful research tools for in vitro studies of GCS and its role in biological processes.

[0029] These and other novel aspects of the invention will become apparent from the following detailed description of the embodiments.

Best Mode for Carrying Out the Invention

[0030] The present invention is described in relation to preferred embodiments. However, it should be recognized that the present invention is not limited to the disclosed embodiments. Considering the description of the embodiments of the present invention herein, it will be understood that various modifications can be made by those skilled in the art. Such modifications are included in the claims set forth below.

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

[0032] The term "disease or condition for which inhibition of GCS provides a benefit" relates to a condition where GCS, and / or the action of GCS, is important or necessary, for example, for the onset, progression, manifestation of that disease or condition, or a disease or condition known to be treated by a GCS inhibitor (eliglustat tartrate). Examples of such conditions include, but are not limited to, Gaucher disease and Fabry disease. One of ordinary skill in the art can readily determine whether a compound treats a disease or condition mediated by GCS, for example, by an assay that can be conveniently used to evaluate the activity of a particular compound.

[0033] The term "second therapeutic agent" refers to a therapeutic agent known to treat a disease or condition of interest that is different from the GCS inhibitor of formula (I). In particular, it can act synergistically with a second therapeutic agent that increases the activity of beta-glucosylceramidase using the GCS inhibitor of formula (I). For example, if Gaucher disease is the disease or condition of interest, the second therapeutic agent can be, for example, isofagamine, enzyme replacement therapy, or gene therapy, etc., which may be known for the treatment of type (I) Gaucher disease or Fabry disease.

[0034] The term "disease" or "condition" is generally considered to be a pathological condition or function, meaning a disorder and / or abnormality that can manifest themselves in the form of specific signs, symptoms, and / or forms of dysfunction. As shown below, the compound of formula (I) is an inhibitor of GCS and can be used for the treatment of diseases and conditions for which inhibition of GCS provides a benefit.

[0035] As used herein, the terms "treat", "treating", "treatment" and the like refer to eliminating, reducing, or alleviating a disease or condition and / or symptoms associated therewith. Without being limiting, treating a disease or condition does not require that the disease, condition, or symptoms associated therewith be completely eliminated. As used herein, the terms "treat", "treating", "treatment" and the like may include "preventive treatment" which refers to reducing the recurrence of a disease or condition, or the probability of recurrence of a disease or condition in a subject who does not have a recurrence of the disease or condition but is at risk of or susceptible to such recurrence, or recurrence of a previously controlled disease or condition. The term "treat" and synonyms are intended to administer a therapeutically effective amount of a compound of the invention to an individual in need of such treatment.

[0036] Within the meaning of the present invention, "treatment" also includes prevention of recurrence or co-prevention, as well as treatment of acute or chronic signs, symptoms and / or dysfunctions. Treatment can be symptomatically adapted, for example, to suppress symptoms. This can be brought about over a short period, adapted over a medium term, or can be long-term treatment, for example, in the context of maintenance therapy.

[0037] As used herein, the term "therapeutically effective amount" or "effective dose" refers to an amount of an active agent that is sufficient to effectively deliver an active agent for the treatment of a condition or disease of interest to an individual in need of treatment when administered by the methods of the present invention. In the case of lysosomal storage diseases, a therapeutically effective amount of the agent reduces (i.e., retards to some extent, preferably halts) the accumulation of unwanted glycolipids and / or alleviates to some extent one or more 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.

[0039] The term "package insert" means the information accompanying a pharmaceutical product that provides instructions on the method of administration of the product, together with safety and efficacy data necessary for a physician, pharmacist, and patient to make a decision based on sufficient information regarding the use of the product. The package insert is generally regarded as the "label" of the pharmaceutical product.

[0040] The terms "co-administration", "administer in combination", "concomitant administration", and similar phrases mean that two or more agents are administered simultaneously to the subject being treated. "Concurrently" means that each agent is administered either simultaneously or sequentially in any order at different times. However, if not administered simultaneously, it means that each agent is administered to the individual in order and at times sufficiently close so as to provide the desired therapeutic effect and be able to act in concert. For example, the GCS inhibitor of structural formula (I) can be administered simultaneously with, or sequentially at different times in any order with, a second therapeutic agent. The present GCS inhibitor and the second therapeutic agent can be administered separately in any suitable form and by any suitable route. It is understood that if the present GCS inhibitor and the second therapeutic agent are not administered simultaneously, they can be administered to the subject in need thereof in any order.

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

[0042] The terms "a", "an", "the", and uses of similar referents in the context of describing the present invention (especially in the context of the claims) are to be construed to cover both the singular and the plural forms unless otherwise specified. The recitation of ranges of values herein is merely intended to serve as a concise way of referring individually to each separate value falling within the range, 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., "such as") provided herein is intended to better illustrate the invention and is not a limitation on the scope of the invention unless otherwise claimed. No language in this specification should be construed as indicating that any non-claimed element is essential for the practice of the invention.

[0043] Compounds that inhibit glycolipid synthesis are known. As such, these compounds can be used in the treatment of diabetes and lysosomal storage diseases such as Tay-Sachs disease, Sandhoff disease, Gaucher disease, Fabry disease, etc. However, to date, these compounds have been limited by low activity, insufficient CNS penetration, or both.

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

[0045] Phase 2 clinical data for eliglustat tartrate showed a clinical response in type 1 Gaucher disease comparable to enzyme replacement therapy, as measured by reduction in spleen and liver volume, correction of anemia, and improvement of thrombocytopenia. Adverse effects observed with NBDNJ, including weight loss, diarrhea, and tremors, were not observed in this clinical trial or in the continuation trial. These observations are consistent with the high specificity of eliglustat tartrate and its lack of CNS penetration. The lack of distribution of eliglustat tartrate into the brain may be advantageous for glycolipid storage disorders without CNS symptoms, including type 1 Gaucher disease and Fabry disease, but the identification of compounds of structural formula (I) that cross the BBB has a therapeutic effect on disorders such as GM2 gangliosidosis, Tay-Sachs disease, Sandhoff disease, and type 2 and 3 Gaucher disease that exhibit CNS symptoms.

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

[0047] A method for preventing the growth of unwanted glycolipid accumulation in a subject, the method comprising administering to a subject having a lysosome with a condition characterized by unwanted glycolipid accumulation a therapeutically effective amount of a compound of structural formula (I). In some embodiments, the compound of structural formula (I) can cross the BBB and is thus useful for the treatment of lysosomal storage diseases, such as type II and type III Gaucher's disease, which were previously not treatable with GCS inhibitors.

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

Chemical formula

[0049] The compounds of structural formula (I) are used in a method of treating a disease or condition in which inhibition of GCS provides a benefit, such as Gaucher disease, Fabry disease, Tay–Sachs disease, Sandhoff disease, diabetes, hypertrophy or hyperplasia associated with diabetic neuropathy, lupus, increased plasma TNF-α, elevated glycated hemoglobin levels, and glomerular diseases. The method comprises administering to an individual in need thereof a therapeutically effective amount of a compound of structural formula (I). The method also encompasses administering to the individual a second therapeutic agent in addition to the compound of structural formula (I). The second therapeutic agent is selected from drugs known to be useful in the treatment of a disease or condition afflicting the individual in need thereof. Such diseases and conditions include, but are not limited to, Parkinson's disease, multiple myeloma, and ADPKD.

[0050] As used herein, the term "C 1~4 alkyl" refers to straight-chain and branched saturated hydrocarbon groups, non-limiting examples of which include methyl, ethyl, straight-chain and branched propyl groups, and straight-chain and branched butyl groups.

[0051] The term "C 3~6 cycloalkyl" refers to saturated cycloalkyl groups containing 3 to 6 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In some embodiments, the -CH2- of the cycloalkyl group is replaced by an oxygen atom.

[0052] C 1~4 alkyl and C 3~6 cycloalkyl groups can be substituted with one or more Fs.

[0053] In various embodiments, R 1 is -H, -OC(CH3)3, -OCH3, -OCF3, -OCHF2,

Chem.

[0054] In another embodiment, R 1 and R 2 together form

Chem.

[0055] In a preferred embodiment, R 3 is H or F.

[0056] In another preferred embodiment, R 4 is H or CH3.

[0057] In another preferred embodiment, R 7 and R 8 one of which is H and the other is OCH3.

[0058] In another preferred embodiment, -NR 5 R 5’ is

Chem.

[0059] In various embodiments,

Chem.

Chem.

[0060] In addition, salts, hydrates, and solvates of the present compounds are also included in the present invention and can be used in the methods disclosed herein. The present invention further encompasses all possible stereoisomers and geometric isomers of the compounds of structural formula (I). The present invention includes both racemic compounds and optically active isomers. When a compound of structural formula (I) is desired as a single enantiomer, it can be obtained either by resolution of the final product or by stereospecific synthesis from either the use of enantiomerically pure starting materials or chiral auxiliaries, see, for example, Z. Ma et al., Tetrahedron: Asymmetry, 8(6), pages 883 - 888 (1997). Resolution of the final product, intermediate, or starting material can be achieved by any suitable method known in the art. In addition, in situations where tautomers of the compounds of structural formula (I) are possible, the present invention is intended to include all tautomeric forms of the compounds.

[0061] The compounds of the present invention can exist as salts. 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 salts" refers to salts or zwitterionic forms of the compounds of structural formula (I). The salts of the compounds of formula (I) can be prepared separately during the final isolation and purification of the compound or by reacting the compound with an acid having a suitable cation. Pharmaceutically acceptable salts of the compounds of structural formula (I) can be acid addition salts formed with pharmaceutically acceptable acids. 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, as well as 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, hydrogen phosphate, acetate, adipate, alginate, aspartate, benzoate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, succinate, fumarate, maleate, ascorbate, isethionate, salicylate, methanesulfonate, mesitylenesulfonate, naphthalenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, trichloroacetate, trifluoroacetate, phosphate, glutamate, glutarate, bicarbonate, paratoluenesulfonate, undecanoate, lactate, citrate, tartrate, gluconate, methanesulfonate, ethanedisulfonate, benzenesulfonate, and p-toluenesulfonate, but are not limited thereto. In addition, available amino groups present in the compounds of the present invention can 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 bromide and phenethyl bromide.In view of the above, any reference in this specification to a compound of the present invention is intended to include the compound of structural formula (I), as well as its pharmaceutically acceptable salts, hydrates, or solvates.

[0062] Some specific embodiments of the present invention include, but are not limited to, the following.

[0063] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Examples]

[0064] Synthesis of Compounds The compounds of the present invention were prepared as follows. The following synthetic schemes represent the reactions used to synthesize the compounds of structural formula (I). Modifications and alternative schemes for preparing the GCS inhibitors of the present invention are readily within the capabilities of those skilled in the art.

[0065] Preparation and Spectral Analysis Data of Compounds of Structural Formula (I) Chemical names follow CAS or IUPAC nomenclature. Starting materials 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 pre-coated silica gel 60F254 plates. Silica gel chromatography was performed using silica gel (220 - 240 mesh) obtained from Silicycle.

[0066] The NMR spectra were recorded on a Bruker 500 MHz spectrometer. Chemical shifts are reported in δ (parts per million) by referencing the hydrogenation residues of the deuterated solvents with the internal standard CDCl3: δ = 7.28( 1 H NMR). The mass spectra were recorded on a Micromass LCT time-of-flight instrument using positive electrospray ionization mode. The purity of the compounds was evaluated by analytical reverse-phase HPLC (Agilent Eclipse Plus C18 4.6×75 mm column (3.5 μm silica), 254 nm detection) using a gradient of 10 - 90% CH3CN / water over 6 minutes.

[0067] Unless otherwise specified, all temperatures are in degrees Celsius.

[0068] In these examples and elsewhere, the abbreviations have the following meanings.

[0069]

Table 2-1

Table 2-2

[0070]

Chem.

[0071] Preparation of (R)-benzyl (3,8,8,9,9-pentamethyl-4-oxo-2,7-dioxa-3-aza-8-siladecane-5-yl)carbamate (3). To a mixture of Compound 1 (5.00 g, 24.4 mmol), Compound 2 (7.15 g, 73.5 mmol), and DIPEA (15.8 g, 122 mmol) in anhydrous CH3CN (60 mL) was added HATU (9.70 g, 25.5 mmol) under N2 at room temperature. The resulting mixture was stirred overnight. Next, the reaction mixture was partitioned between water and EtOAc. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was dried under high vacuum and then dissolved in THF (60 mL). To the resulting solution were added imidazole (2.99 g, 43.9 mmol), followed by TBDMSCl (5.50 g, 36.5 mmol). The reaction mixture was stirred at room temperature overnight. Next, the reaction mixture was concentrated under reduced pressure. The residue was partitioned between water and EtOAc. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 10% to 30% EtOAc / hexane to afford Compound 3 as a clear oil (7.10 g, 73%): 1 1H NMR (500 MHz, CDCl3) δ 7.33 - 7.28 (m, 5H), 5.60 (d, J = 8.5 Hz, 1H), 5.08 (ABq, J = 12.3 Hz, 2H), 4.78 (br s, 1H), 3.88 - 3.78 (m, 2H), 3.74 (s, 3H), 3.20 (s, 3H), 0.84 (s, 9H), 0.01 (s, 6H).

[0072] Preparation of (R)-benzyl (3-((tert-butyldimethylsilyl)oxy)-1-(3-chlorophenyl)-1-oxopropan-2-yl)carbamate (5). To a stirred mixture of Compound 4 (5.59 g, 23.4 mmol) in anhydrous THF (100 mL) was added 2 M iA PrMgCl solution was added dropwise. The resulting mixture was stirred for 30 minutes. To the resulting mixture, a solution of compound 3 (7.20 g, 18.2 mmol) in anhydrous THF (30 mL) was added. The reaction mixture was stirred at room temperature for 1 hour, then cooled to -20 °C, quenched with saturated aqueous NH4Cl, and extracted with EtOAc. 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 4% to 16% EtOAc / hexane to give compound 5 as a clear oil (7.58 g, 93%): ESI MS, m / z = 448 [M+H] + 。

[0073] Preparation of benzyl ((1R,2R)-3-((tert-butyldimethylsilyl)oxy)-1-(3-chlorophenyl)-1-hydroxypropan-2-yl)carbamate (6). To a stirred solution of compound 5 (7.58 g, 16.9 mmol) in anhydrous THF (80 mL) was added dropwise a 1 M solution of L-selectride in THF (33.8 mL, 33.8 mmol) at -78 °C under N2. The resulting mixture was stirred at -78 °C for 1 hour, then quenched with saturated aqueous NH4Cl and extracted with EtOAc. 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 8% to 30% EtOAc / hexane to give compound 6 as a clear oil (7.11 g, 93%): ESI MS, m / z = 450 [M+H] + 。

[0074] Preparation of benzyl ((1R,2R)-1-(3-chlorophenyl)-1,3-dihydroxypropan-2-yl)carbamate (7). A solution of compound 6 (7.11 g, 15.8 mmol) in HOAc (40 mL), THF (20 mL), and water (20 mL) was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was dissolved in EtOAc and washed with water. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 20% to 100% EtOAc / hexane to afford compound 7 as a clear syrup (4.92 g, 93%): ESI MS, m / z = 336 [M+H] + .

[0075] Preparation of (2R,3R)-2-(((benzyloxy)carbonyl)amino)-3-(3-chlorophenyl)-3-hydroxypropyl 4-methylbenzenesulfonate (8). To a stirred solution of compound 7 (4.92 g, 14.7 mmol), Et3N (4.43 g, 43.8 mmol), and DMAP (179 mg, 1.47 mmol) in anhydrous CH2Cl2 (40 mL) at room temperature under N2 was added dropwise over 2 h (using a syringe pump) a solution of TsCl (3.07 g, 16.1 mmol) in anhydrous CH2Cl2 (120 mL). The reaction mixture was stirred for 1 h and then concentrated under reduced pressure (bath temperature < 30 °C). The resulting residue was purified directly by flash column chromatography on silica gel eluting with 10% to 40% EtOAc / hexane to afford compound 8 as a white foam (6.69 g, 93%): ESI MS, m / z = 490 [M+H] + .

[0076] Preparation of benzyl ((1R,2R)-1-(3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)carbamate (9). A solution of compound 8 (6.69 g, 13.7 mmol) and pyrrolidine (4.86 g, 68.3 mmol) in anhydrous CH3CN (80 mL) was heated in an oil bath at 75 °C for 2 h, then cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 2% to 20% MeOH / CH2Cl2 to afford compound 9 as a pale yellow foam (2.85 g, 54%): ESI MS, m / z = 389 [M+H] + .

[0077] Preparation of (1R,2R)-2-amino-1-(3-chlorophenyl)-3-(pyrrolidin-1-yl)propan-1-ol (10). To a stirred solution of compound 9 (2.85 g, 7.33 mmol) in HOAc (75 mL) was added aqueous 33% HBr in HOAc (15 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h and then concentrated under reduced pressure. The resulting residue was basified to pH 9 with 6N aqueous NaOH using an ice / water cooling bath and then, i extracted with PrOH / CHCl3 (1:10) (monitoring the aqueous phase by LC / MS). The combined extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting residue was purified by a C18 column eluting with 0% to 100% CH3CN / H2O (containing 0.05% TFA). The fractions containing the desired product were combined and concentrated under reduced pressure. The residue was basified with saturated aqueous NaHCO3 and, i extracted with PrOH / CHCl3 (1:10) (monitoring the aqueous phase by LC / MS). The combined extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford compound 10 as a clear oil (910 mg, 49%): ESI MS, m / z = 255 [M+H] + .

[0078] Preparation of 2,5-dioxopyrrolidin-1-yl 2-(2,3-dihydro-1H-inden-2-yl)acetate (11) To a solution of N-hydroxysuccinimide (1.94 g, 17.0 mmol) in CH2Cl2 (100 mL) were added 2-(2,3-dihydro-1H-inden-2-yl)acetic acid (3.00 g, 17.0 mmol) and EDC (3.58 g, 18.0 mmol). The resulting solution was stirred at room temperature for 18 h. The reaction mixture was diluted with saturated sodium bicarbonate (100 mL). The layers were separated and the aqueous phase was extracted with CH2Cl2. The combined organic extracts were washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound 11 as an off-white solid (4.40 g, 94%): ESI MS m / z 296 [M+Na] + .

[0079] Preparation of N-((1R,2R)-1-(3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 1). A solution of compound 10 (910 mg, 3.57 mmol), compound 11 (1.07 g, 3.92 mmol) and Et3N (1.09 g, 10.8 mmol) in anhydrous CH2Cl2 (50 mL) was stirred at room temperature overnight. The reaction mixture was diluted with CH2Cl2, washed with saturated aqueous NaHCO3, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 4% to 30% MeOH / CH2Cl2 and further purified by C18 column eluting with 0% to 100% CH3CN / H2O (containing 0.05% TFA). The fractions containing the desired product were combined and concentrated under reduced pressure. The residue was basified with saturated aqueous NaHCO3 and extracted with CH2Cl2. The combined extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound Example 1 as a white solid (884 mg, 60%): 11H NMR (500 MHz, CDCl3) δ 7.40 - 7.25 (m, 3H), 7.21 - 7.11 (m, 5H), 5.88 (d, J = 7.5 Hz, 1H), 5.07 (d, J = 2.7 Hz, 1H), 4.26 - 4.23 (m, 1H), 3.00 - 2.63 (m, 9H), 2.49 (dd, J = 15.6, 6.7 Hz, 1H), 2.33 (dd, J = 15.6, 6.6 Hz, 1H), 2.28 (dd, J = 14.1, 7.1 Hz, 1H), 2.18 (dd, J = 14.1, 8.1 Hz, 1H), 1.85 - 1.75 (m, 4H); ESI MS, m / z = 413 [M + H] + 。

[0080] The following compound was prepared as in typical Scheme 1 using the commercially available aryl iodide required in place of Compound 4.

Chemical formula

[0081] N-((1R,2R)-1-(4-(tert-Butoxy)-3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 2). 1 1H NMR (300 MHz, DMSO-d6) δ 7.55 - 7.52 (m, 1H), 7.40 (d, J = 1.8 Hz, 1H), 7.19 - 7.06 (m, 6H), 5.80 (bs, 1H), 4.81 (s, 1H), 4.26 - 4.05 (m, 1H), 2.90 - 2.65 (m, 3H), 2.60 - 2.40 (m, 5H), 2.39 - 2.05 (m, 4H), 1.68 (bs, 4H), 1.32 (s, 9H); ESI MS, m / z = 485 [M + H] + 。

Chemical formula

[0082] N-((1R,2R)-1-(3-chloro-4-(trifluoromethoxy)phenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 3). 1 1H NMR (500 MHz, CDCl3) δ 7.53 (d, J = 1.9 Hz, 1H), 7.32 - 7.28 (m, 1H), 7.27 - 7.23 (m, 1H), 7.15 - 7.08 (m, 4H), 5.99 - 5.90 (m, 1H), 5.11 - 5.07 (m, 1H), 4.30 - 4.24 (m, 1H), 3.04 - 2.96 (m, 2H), 2.95 - 2.83 (m, 2H), 2.82 - 2.62 (m, 5H), 2.54 - 2.44 (m, 1H), 2.37 - 2.24 (m, 2H), 2.22 - 2.13 (m, 1H), 1.91 - 1.77 (m, 4H); ESI MS m / z 497 [M+H] + . [Chemical Structure]

[0083] N-((1R,2R)-1-(3-chlorocyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 4). 1 1H NMR (500 MHz, CDCl3) δ 7.38 - 7.37 (m, 1H), 7.25 - 7.11 (m, 6H), 5.89 (d, J = 7.6 Hz, 1H), 5.01 (d, J = 2.8 Hz, 1H), 4.26 - 4.20 (m, 1H), 3.80 - 3.75 (m, 1H), 2.99 (dd, J = 15.6, 7.7 Hz, 1H), 2.94 - 2.83 (m, 3H), 2.81 - 2.62 (m, 5H), 2.51 (dd, J = 15.6, 6.7 Hz, 1H), 2.37 (dd, J = 15.6, 6.6 Hz, 1H), 2.30 (dd, J = 14.0, 7.0 Hz, 1H), 2.20 (dd, J = 14.1, 8.1 Hz, 1H), 1.85 - 1.75 (m, 4H), 0.85 - 0.75 (m, 4H); ESI MS, m / z = 469 [M+H] + . [Chemistry]

[0084] N-((1R,2R)-1-(3-chloro-4-(difluoromethoxy)phenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 13). 1 H NMR (500 MHz, CDCl3) δ 7.50~7.49 (m, 1H), 7.24~7.20 (m, 2H), 7.16~7.10 (m, 4H), 6.49 (t, J = 73.5 Hz, 1H), 5.92 (d, J = 7.6 Hz, 1H), 5.06 (d, J = 2.6 Hz, 1H), 4.26~4.22 (m, 1H), 3.02~2.95 (m, 2H), 2.91~2.85 (m, 2H), 2.78~2.62 (m, 5H), 2.50 (dd, J = 15.6, 6.7 Hz, 1H), 2.35 (dd, J = 15.6, 6.6 Hz, 1H), 2.29 (dd, J = 14.2, 7.1 Hz, 1H), 2.19 (dd, J = 14.2, 8.0 Hz, 1H), 1.85~1.76 (m, 4H); ESI MS, m / z = 479 [M+H] + . [Chemistry]

[0085] N-((1R,2R)-1-(3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(5-fluoro-2,3-dihydro-1H-inden-2-yl)acetamide (Example 9). Prepared as in representative Scheme 1, using 2-(5-fluoro-2,3-dihydro-1H-inden-2-yl)acetic acid instead of 2-(2,3-dihydro-1H-inden-2-yl)acetic acid. 11H NMR (500 MHz, DMSO-d6) δ 7.52 (bs, 1H), 7.39 (s, 1H), 7.35 - 7.23 (m, 3H), 7.12 (m, 1H), 6.96 - 6.86 (m, 2H), 5.72 (bs, 1H), 4.86 (s, 1H), 4.12 (bs, 1H), 2.76 - 2.68 (m, 2H), 2.67 - 2.23 (m, 8H), 2.18 - 2.05 (m, 2H), 1.68 (bs, 4H); ESI MS, m / z = 431 [M+H] + 。

Chem.

[0086] N-((1R,2R)-1-(4-(tert-Butyl)-3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 21). ESI MS, m / z = 469.2 [M+H] + 。

Chem.

[0087] N-((1R,2R)-1-(3-Chlorophenyl)-3-((R)-3-fluoropyrrolidin-1-yl)-1-hydroxypropyl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 10). Prepared as in representative Scheme 1 using (R)-3-fluoropyrrolidine instead of pyrrolidine. 1 1H NMR (300 MHz, CDCl3) δ 7.40 (s, 1H), 7.32 - 7.24 (m, 2H), 7.23 - 7.10 (m, 5H), 5.99 (d, J = 7.2 Hz, 1H), 5.28 - 5.05 (m, 2H), 4.25 - 4.18 (m, 1H), 3.18 - 2.67 (m, 8H), 2.65 - 2.46 (m, 2H), 2.38 - 1.96 (m, 5H); ESI MS, m / z = 431 [M+H] + 。

Chem.

[0088] N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-3-((R)-3-fluoropyrrolidin-1-yl)-1-hydroxypropan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 11). Prepared as described for Example 4 using (R)-3-fluoropyrrolidine instead of pyrrolidine. 1 H NMR (500 MHz, CDCl3) δ 7.38 (d, J = 2.1 Hz, 1H), 7.38 - 7.25 (m, 1H), 7.19 - 7.10 (m, 5H), 5.98 (d, J = 6.6 Hz, 1H), 5.23 - 5.10 (m, 1H), 4.99 (d, J = 3.1 Hz, 1H), 4.23 - 4.17 (m, 1H), 3.79 - 3.75 (m, 1H), 3.14 - 2.98 (m, 3H), 2.92 - 2.49 (m, 7H), 2.39 (dd, J = 15.6, 6.7 Hz, 1H), 2.32 (dd, J = 14.1, 7.1 Hz, 1H), 2.23 (dd, J = 14.1, 8.0 Hz, 1H), 2.19 - 2.04 (m, 2H), 0.83 - 0.79 (m, 4H); ESI MS, m / z = 487 [M + H] + 。

[0089] N-((1R,2R)-3-(azetidin-1-yl)-1-(3-chlorophenyl)-1-hydroxypropan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 16) Prepared as in Scheme 1 using azetidine instead of pyrrolidine. 11H NMR (499 MHz, chloroform-d) δ 7.40 (s, 1H), 7.32 - 7.23 (m, 2H), 7.23 - 7.16 (m, 1H), 7.13 (br s, 4H), 5.81 (d, J = 7.8 Hz, 1H), 5.04 (d, J = 2.1 Hz, 1H), 4.12 (dq, J = 6.6, 3.2 Hz, 1H), 3.36 (dq, J = 23.1, 7.1 Hz, 4H), 3.02 (dd, J = 12.8, 3.9 Hz, 1H), 2.95 (dd, J = 15.6, 7.7 Hz, 1H), 2.85 (dd, J = 12.8, 3.2 Hz, 1H), 2.79 (dd, J = 15.4, 7.7 Hz, 1H), 2.71 (hept, J = 7.1 Hz, 1H), 2.47 (dd, J = 15.6, 6.8 Hz, 1H), 2.30 - 2.20 (m, 2H), 2.19 - 2.06 (m, 3H); HRMS (ESI) m / z: [M + H] + Calcd for 399.1839; Found: 399.1832.

[0090] The following compounds were prepared as in representative Scheme 1 using non-commercial aryl iodides instead of Compound 4. The syntheses of these iodides are included below.

[0091] N-((1R,2R)-1-(3-chloro-4-((3-methyloxetan-3-yl)oxy)phenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 5). Prepared as in Scheme 1 using 3-(2-chloro-4-iodophenoxy)-3-methyloxetane. 1 ​1H NMR (500 MHz, CDCl3) δ 7.44~7.43 (m, 1H), 7.17~7.08 (m, 5H), 6.43 (d, J = 8.5 Hz, 1H), 5.91 (d, J = 7.5 Hz, 1H), 5.00 (d, J = 2.8 Hz, 1H), 4.96 (d, J = 6.6 Hz, 2H), 4.55 (d, J = 7.2 Hz, 2H), 4.26~4.20 (m, 1H), 3.01 (dd, J = 15.6, 7.7 Hz, 1H), 2.94~2.83 (m, 3H), 2.81~2.61 (m, 5H), 2.52 (dd, J = 15.6, 6.6 Hz, 1H), 2.38 (dd, J = 15.6, 6.6 Hz, 1H), 2.30 (dd, J = 14.1, 7.1 Hz, 1H), 2.20 (dd, J = 14.1, 8.1 Hz, 1H), 1.85~1.75 (m, 4H), 1.73 (s, 3H); ESI MS, m / z = 499 [M+H] + 。

Chem.

[0092] N-((1R,2R)-1-(8-chloro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 7). Prepared as in Scheme 1 using 8-chloro-6-iodo-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin. 11H NMR (500 MHz, CDCl3) δ 7.17~7.11 (m, 4H), 6.97~6.96 (m, 1H), 6.80~6.79 (m, 1H), 5.88 (d, J = 7.4 Hz, 1H), 4.94 (d, J = 2.9 Hz, 1H), 4.20~4.17 (m, 1H), 3.84 (ABq, J = 11.1 Hz, 2H), 3.01 (dd, J = 15.6, 7.9 Hz, 1H), 2.94~2.85 (m, 3H), 2.80~2.61 (m, 5H), 2.52 (dd, J = 15.6, 6.7 Hz, 1H), 2.41 (dd, J = 15.6, 6.7 Hz, 1H), 2.30 (dd, J = 14.1, 7.1 Hz, 1H), 2.22 (dd, J = 14.0, 8.1 Hz, 1H), 1.83~1.75 (m, 4H), 1.37 (s, 6H); ESI MS, m / z = 499 [M+H] + 。

Chem.

[0093] N-((1R,2R)-1-(3-chloro-4-cyclopropylphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 8). Prepared as in Scheme 1 using 3-chloro-4-cyclopropyl-1-iodobenzene. 1 1H NMR (500 MHz, DMSO-d6) δ 7.52~7.48 (m, 1H), 7.36 (s, 1H), 7.18~7.05 (m, 5H), 6.93 (d, J = 8.5 Hz, 1H), 5.65 (bs, 1H), 4.81 (s, 1H), 4.10 (bs, 1H), 2.78~2.72 (m, 2H), 2.67~2.45 (m, 4H), 2.41 (dd, J = 15.5, 7.5 Hz, 1H), 2.30~2.25 (m, 3H), 2.18~2.11 (m, 3H), 1.68 (bs, 4H), 0.98 (m, 2H), 0.65 (m, 2H); ESI MS, m / z = 453 [M+H] + 。

[0094] Preparation of tert-butyl (4-bromo-3-chlorophenyl)carbamate [Chemistry] To a stirred solution of compound 4-bromo-3-chloroaniline (19.5 g, 94.4 mmol) in water (200 mL) was added Boc2O (30.9 g, 142 mmol) at room temperature. The resulting mixture was stirred at room temperature overnight. Next, the reaction mixture was extracted with EtOAc (3×). The organic layer was 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 20% EtOAc / hexane to afford compound 2 as a purple solid (21.5 g, 63%): 1 1H NMR (300 MHz, DMSO-d6) δ 9.71 (br s, 1H), 7.78 (d, J = 2.3 Hz, 1H), 7.62 (d, J = 8.8 Hz, 1H), 7.31 (dd, J = 8.8, 2.4 Hz, 1H), 1.47 (s, 9H).

[0095] Preparation of tert-butyl (3-chloro-4-cyclopropylphenyl)carbamate [Chemistry] A mixture of tert-butyl (4-bromo-3-chlorophenyl)carbamate (620 mg, 2.02 mmol), cyclopropyltrifluoro-λ 4 -borane, potassium salt (578 mg, 4.04 mmol), P(Cy)3 (113 mg, 0.403 mmol), and K3PO4 (1.50 g, 7.07 mmol) in water (5 mL) and toluene (10 mL) in a sealed tube was bubbled with argon for 5 minutes. To the resulting mixture was added Pd(OAc)2 (45 mg, 0.200 mmol) at room temperature. The resulting mixture was sealed and heated at 110 °C overnight. Thereafter, the reaction mixture was cooled to room temperature and extracted with EtOAc (3×). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 0.5% to 2% EtOAc / hexane to afford the title compound as a yellow oil (370 mg, 68%): 11H NMR (500 MHz, DMSO-d6) δ 9.42 (br s, 1H), 7.58 (d, J = 1.3 Hz, 1H), 7.24 (dd, J = 8.5, 2.1 Hz, 1H), 6.91 (d, J = 8.5 Hz, 1H), 1.98 - 2.08 (m, 1H), 1.47 (s, 9H), 0.94 - 0.91 (m, 2H), 0.62 - 0.60 (m, 2H).

[0096] Preparation of 3-Chloro-4-cyclopropylaniline

Chem.

[0097] Preparation of 2-Chloro-1-cyclopropyl-4-iodobenzene

Chem.

[0098] Preparation of diethyl 2-(4-bromo-2-chlorophenoxy)malonate

Chemical formula

[0099] Preparation of Diethyl 2-(4-Bromo-2-chlorophenoxy)-2-methylmalonate

Chem.

[0100] Preparation of 2-(4-Bromo-2-chlorophenoxy)-2-methylpropane-1,3-diol

Chem.

[0101] Preparation of 3-(4-bromo-2-chlorophenoxy)-3-methyloxetane

Chemical formula

[0102] Preparation of 3-(2-chloro-4-iodophenoxy)-3-methyloxetane [Chemical formula] A stirred suspension of 3-(4-bromo-2-chlorophenoxy)-3-methyloxetane (5.78 g, 20.8 mmol), trans-cyclohexane-1,2-diamine (325 mg, 2.28 mmol), NaI (6.24 g, 41.6 mmol), and CuI (198 mg, 1.04 mmol) in anhydrous 1,4-dioxane (50 mL) was heated to reflux overnight (115 °C oil bath) under nitrogen. The reaction mixture was then cooled to room temperature, diluted with EtOAc (50 mL) and hexane (100 mL), and filtered. The filter cake was washed with EtOAc (50 mL). The filtrate was washed with saturated aqueous NH4Cl (2 × 100 mL), 20% aqueous Na2S2O3 (100 mL), and brine (200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 4% to 20% EtOAc / hexane to afford the title compound as a clear syrup (6.47 g, 96%): 1 H NMR (300 MHz, CDCl3) δ 7.70 (d, J = 2.1 Hz, 1H), 7.43 (dd, J = 8.6, 2.1 Hz, 1H), 6.19 (d, J = 8.6 Hz, 1H), 4.95 (d, J = 6.6 Hz, 2H), 4.57 (d, J = 7.3 Hz, 2H), 1.74 (s, 3H).

[0103] Preparation of 5-bromo-1-chloro-3-fluoro-2-(methoxymethoxy)benzene

Chemical formula

[0104] Preparation of 5-bromo-1-chloro-2-(methoxymethoxy)-3-((2-methylallyl)oxy)benzene and 5-bromo-1-chloro-2-(methoxymethoxy)-3-((2-methylprop-1-en-1-yl)oxy)benzene [Chemical formula] To a stirred solution of 2-methylprop-2-en-1-ol (3.86 g, 53.5 mmol) in anhydrous DMF (50 mL) was added NaH (60%, 2.13 g, 53.3 mmol) portionwise at 0 °C under nitrogen. The resulting mixture was stirred at 0 °C for 30 minutes. To the reaction mixture was added a solution of 5-bromo-1-chloro-3-fluoro-2-(methoxymethoxy)benzene [7.26 g (crude), 26.6 mmol (approx.)] in anhydrous DMF (8 mL). The resulting mixture was warmed to room temperature and stirred over the weekend. Thereafter, the reaction mixture was quenched with ice-cold water (200 mL) and extracted with 1:1 EtOAc / hexane (3 × 100 mL). The combined extracts were washed with 10% aqueous LiCl (2 × 50 mL) and brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel eluting with 1% to 2% EtOAc / hexane to afford the title compound as a clear oil (7.45 g, 86%): 1 H NMR (300 MHz, CDCl3) δ 7.19 (d, J = 2.3 Hz, 0.2H), 7.15 (d, J = 2.2 Hz, 1H), 6.99 (d, J = 2.3 Hz, 0.2H), 6.93 (d, J = 2.2 Hz, 1H), 6.08 (s, 0.2H), 5.18 (s, 0.4H), 5.16 (s, 2H), 5.08 (s, 1H), 5.02 (s, 1H), 4.43 (s, 2H), 3.65 (s, 0.6H), 3.62 (s, 3H), 1.76 (s, 3H), 1.71 (s, 0.6H), 1.70 (s, 0.6H).

[0105] Preparation of 4-bromo-2-chloro-6-((2-methylallyl)oxy)phenol and 4-bromo-2-chloro-6-((2-methylprop-1-en-1-yl)oxy)phenol

Chemical formula

[0106] Preparation of 6-bromo-8-chloro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin

Chemical formula

[0107] Preparation of 8-chloro-6-iodo-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine

Chemical Structure

[0108] Examples 22 and 23 are prepared as described in representative Scheme 1 starting from appropriately substituted aryl iodides.

[0109] [Chemical formula] N-((1R,2R)-1-(3-chloro-2-fluorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 22) 11H NMR (400 MHz, CDCl3) δ 7.41 (t, J = 6.9 Hz, 1H), 7.33 (t, J = 6.9 Hz, 1H), 7.19 - 7.04 (m, 5H), 6.04 (d, J = 7.8 Hz, 1H), 5.39 (s, 1H), 4.33 - 4.20 (m, 1H), 3.02 - 2.83 (m, 4H), 2.83 - 2.60 (m, 5H), 2.51 (dd, J = 15.6, 6.7 Hz, 1H), 2.37 (dd, J = 15.6, 6.7 Hz, 1H), 2.24 (qd, J = 14.1, 7.5 Hz, 2H), 1.88 - 1.72 (m, 4H); HRMS (ESI) m / z: [M+H] + Calculated value: 431.1902; Measured value: 431.1887.

[0110]

Chemical Structure

Chemical Structure

[0111] (4R,5R)-4-(((tert-Butyldimethylsilyl)oxy)methyl)-5-(3-chlorophenyl)oxazolidin-2-one (12) To a solution of benzyl ((1R,2R)-3-((tert-butyldimethylsilyl)oxy)-1-(3-chlorophenyl)-1-hydroxypropan-2-yl)carbamate 6 (2.00 g, 4.44 mmol) in THF (20 mL) was added a 60% NaH dispersion in oil (210 mg, 5.33 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 h. Saturated aqueous ammonium chloride solution was added to quench the reaction, diluted with water, and extracted twice with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered, and concentrated. The crude residue was purified by flash chromatography eluting with 0-50% EtOAc in hexane to afford the title compound as a colorless oil which solidified on standing. (1.21 g, 80%)

[0112] (4R,5R)-5-(3-Chlorophenyl)-4-(hydroxymethyl)oxazolidin-2-one (13). To a solution of (4R,5R)-4-(((tert-butyldimethylsilyl)oxy)methyl)-5-(3-chlorophenyl)oxazolidin-2-one 12 (1.00 g, 2.92 mmol) in acetonitrile (20 mL) was added 48% aqueous HF (1.80 mL, 49.7 mmol). After 1 h, the reaction mixture was concentrated and purified by column chromatography eluting with 0-10% methanol in dichloromethane to afford the title compound as a clear colorless oil. (606 mg, 91%)

[0113] ((4R,5R)-5-(3-Chlorophenyl)-2-oxooxazolidin-4-yl)methyl 4-methylbenzenesulfonate (14) A solution of (4R,5R)-5-(3-chlorophenyl)-4-(hydroxymethyl)oxazolidin-2-one 13 (426 mg, 1.87 mmol) in pyridine (2.5 mL) was treated with 4-methylbenzenesulfonyl chloride (535 mg, 2.81 mmol). The reaction was stirred at room temperature for 5 h and then concentrated. The crude residue was purified by column chromatography eluting with 0-10% methanol in dichloromethane to afford the title compound, which was crystallized from dichloromethane / hexanes to give the title compound as a white solid. (457 mg, 64%).

[0114] (4R,5R)-5-(3-chlorophenyl)-4-(pyrrolidin-1-ylmethyl)oxazolidin-2-one (15) A solution of ((4R,5R)-5-(3-chlorophenyl)-2-oxooxazolidin-4-yl)methyl 4-methylbenzenesulfonate (591 mg, 1.55 mmol) in acetonitrile (8 mL) was treated with pyrrolidine (0.64 mL, 7.7 mmol). The reaction was equipped with a reflux condenser, heated to 75 °C and stirred for 4 h. The reaction was concentrated. The crude residue was purified by column chromatography eluting with 0-10% methanol in dichloromethane to afford the title compound as a colorless film, which solidified on standing. (327 mg, 75%)

[0115] (1R,2R)-2-amino-1-(3-chlorophenyl)-3-(pyrrolidin-1-yl)propan-1-ol (10). A solution of (4R,5R)-5-(3-chlorophenyl)-4-(pyrrolidin-1-ylmethyl)oxazolidin-2-one 15 (272 mg, 0.969 mmol) in ethanol (5.0 mL) was treated with potassium hydroxide (2 M aqueous solution 2 mL, 4.84 mmol). The reaction was heated at 70 °C for 16 h in a sealed tube. The reaction was cooled to room temperature, then diluted with dichloromethane and washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated. The crude residue was purified by column chromatography eluting with 3 - 7% (7N NH3 in methanol) / dichloromethane to afford the title compound as a clear colorless gum. (211 mg, 86%).

[0116] 2-Methyl-2,3-dihydro-1H-inden-2-yl ((1R,2R)-1-(3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)carbamate (Example 14) A solution of (1R,2R)-2-amino-1-(3-chlorophenyl)-3-(pyrrolidin-1-yl)propan-1-ol 10 (50 mg, 0.20 mmol) in dichloroethane (1.5 mL) was treated with 2-methyl-2,3-dihydro-1H-inden-2-yl (4-nitrophenyl) carbonate (prepared from 2-methyl-2-indanol as described below for 2,3-dihydro-1H-inden-2-yl (4-nitrophenyl) carbonate) (80 mg, 0.26 mmol), followed by the addition of triethylamine (54 μL, 0.39 mmol). The reaction was heated at 60 °C for 16 h. Another 30 mg of 2-methyl-2,3-dihydro-1H-inden-2-yl (4-nitrophenyl) carbonate (prepared as described below for 2,3-dihydro-1H-inden-2-yl (4-nitrophenyl) carbonate) (30 mg, 0.96 mmol) was added along with additional triethylamine (50 μL, 0.33 mmol). After 4 h, the reaction was diluted with dichloromethane and washed with brine. The organic layer was dried over magnesium sulfate, filtered, and concentrated. The crude residue was purified by column chromatography eluting with 0–10% methanol in dichloromethane to afford the title compound as an amorphous solid (24 mg, 29%). 1 H NMR (400 MHz, chloroform-d) δ 7.36 (s, 1H), 7.29–7.19 (m, 2H), 7.17 - 7.15 (m, 5H), 5.08–4.98 (m, 1H), 4.89 (d, J = 8.6 Hz, 1H), 3.99 (m, 1H), 3.31 (dd, J = 41.2, 16.5 Hz, 2H), 3.06 (dd, J = 16.4, 10.9 Hz, 2H), 2.84 (ddd, J = 41.6, 12.9, 5.1 Hz, 2H), 2.67 (m, 4H), 1.72 (brs, 4H), 1.56 (s, 3H); HRMS (ESI) m / z: [M + H] + Calculated 429.1945; Found: 429.1939.

[0117] Preparation of 2,3-dihydro-1H-inden-2-yl (4-nitrophenyl) carbonate To a stirred solution of 2-indanol (1.34 g, 10 mmol) in anhydrous CH2Cl2 (50 mL) at 0 °C under nitrogen was added 4-nitrophenyl chloroformate (2.01 g, 10 mmol), followed by a solution of pyridine (0.81 mL, 10 mmol) in anhydrous CH2Cl2 (10 mL). After the addition was complete, the reaction mixture was warmed to room temperature and stirred overnight. The reaction mixture was then concentrated under reduced pressure. The residue was dissolved in EtOAc (50 mL), washed with 10% aqueous citric acid (2 × 40 mL) and brine (40 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with EtOAc and dried under high vacuum to give Compound 3 as an off-white solid (1.17 g, 39%): 1 1H NMR (500 MHz, CDCl3) δ 8.29 - 8.26 (m, 2H), 7.39 - 7.21 (m, 6H), 5.59 - 5.55 (m, 1H), 3.41 (dd, J = 17.0, 6.5 Hz, 2H), 3.20 (dd, J = 17.0, 2.5 Hz, 2H).

[0118] The following compounds were prepared as shown in Scheme 2 starting from the required aryl iodide, amine, and 4-nitrophenyl carbonate.

[0119] 2,3-Dihydro-1H-inden-2-yl ((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)carbamate (Example 12) 1 1H NMR (500 MHz, CDCl3) δ 7.33 - 7.15 (m, 7H), 5.40 - 5.37 (m, 1H), 5.07 (s, 1H), 5.02 (br s, 1H), 4.05 (br s, 1H), 3.80 - 3.77 (m, 1H), 3.30 - 2.60 (m, 10H), 1.89 (br s, 4H), 0.85 - 0.75 (m, 4H); ESI MS, m / z = 471 [M + H] + 。

[0120] 2,3-Dihydro-1H-inden-2-yl ((1R,2R)-1-(3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl) carbamate (Example 6) 1 H NMR (500 MHz, CDCl3) δ 7.34 (s, 1H), 7.23 - 7.04 (m, 7H), 6.80 (bs, 1H), 5.40 - 5.37 (m, 1H), 5.03 (s, 1H), 4.89 - 4.87 (m, 1H), 4.02 (bs, 1H), 3.27 - 3.18 (m, 2H), 2.97 - 2.93 (m, 1H), 2.88 - 2.55 (m, 7H), 1.79 (bs, 4H); ESI MS, m / z = 415 [M+H] + 。

[0121] 2,3-Dihydro-1H-inden-2-yl ((1R,2R)-1-(3-chlorophenyl)-3-((R)-3-fluoropyrrolidin-1-yl)-1-hydroxypropan-2-yl) carbamate oxalate (Example 15) 1 H NMR (400 MHz, DMSO-d6) δ 7.37 - 7.07 (m, 8H), 6.80 (d, J = 9.4 Hz, 1H), 5.33 (d, J = 54.4 Hz, 1H), 5.13 (m, 1H), 4.75 (br s, 1H), 3.98 (m, 1H), 3.39 - 2.51 (m, 10H), 2.31 - 1.90 (m, 2H); HRMS (ESI) m / z: [parent + H] + Calculated value 433.1694; Measured value: 433.1683.

Chemical Structure

[0122] N-((1R,2R)-1-(3-chlorophenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)-N-(methyl-d3)acetamide (Example 17). Prepared as in Scheme 2 using (4R,5R)-4-(((tert-butyldimethylsilyl)oxy)methyl)-5-(3-chlorophenyl)-3-(methyl-d3)oxazolidin-2-one instead of Compound 12, and the synthesis is shown below.

[0123] 1 H NMR (400 MHz, CDCl3) (peaks of the reported major rotamer) 7.38 (t, J = 1.9 Hz, 1H), 7.25 - 7.16 (m, 5H), 7.16 - 7.10 (m, 2H), 4.93 (d, J = 5.2 Hz, 1H), 3.18 - 3.07 (m, 2H), 3.02 - 2.70 (m, 3H), 2.68 - 2.47 (m, 7H), 2.38 - 2.30 (m, 2H), 1.84 - 1.71 (m, 4H). δHRMS (ESI) m / z: [M+H] + Calculated value 430.2341; Measured value: 430.2327.

Chemical Structure

[0124] (4R,5R)-4-(((tert-butyldimethylsilyl)oxy)methyl)-5-(3-chlorophenyl)-3-(methyl-d3)oxazolidin-2-one To a solution of (4R,5R)-4-(((tert-butyldimethylsilyl)oxy)methyl)-5-(3-chlorophenyl)oxazolidin-2-one (1.20 g, 3.51 mmol) in DMF (15 mL) was slowly added sodium hydride (60% oil dispersion) (0.21 g, 5.26 mmol) at 0 °C. After stirring for 1 hour, iodomethane-d3 (2.54 g, 1.10 mL, 5 equiv, 17.5 mmol) was added. After 2 hours, the reaction was quenched by adding saturated ammonium chloride solution and extracted with dichloromethane (3×). The combined organic layers were dried over magnesium sulfate, filtered, and concentrated. Purification by column chromatography eluting with 0 - 50% ethyl acetate in hexane gave the title compound as a solid on standing and isolated as a colorless transparent oil (1.27 g, 100%).

Chemical formula

[0125] N-((1R,2R)-1-(3-chloro-2-methoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 18) 1 H NMR (400 MHz, CDCl3) δ 7.40 (dd, J = 7.8, 1.6 Hz, 1H), 7.31 (dd, J = 8.0, 1.6 Hz, 1H), 7.16 - 7.09 (m, 4H), 7.06 (t, J = 7.8, 6.9 Hz, 1H), 6.32 (d, J = 7.6 Hz, 1H), 5.36 (d, J = 2.8 Hz, 1H), 4.33 (m, 1H), 3.92 (s, 3H), 2.99 - 2.84 (m, 4H), 2.85 - 2.69 (m, 5H), 2.51 (dd, J = 15.5, 6.9 Hz, 1H), 2.40 (dd, J = 15.6, 6.9 Hz, 1H), 2.34 - 2.15 (m, 2H), 1.93 - 1.74 (m, 4H).

Chemical formula

[0126] N-((1R,2R)-1-(5-chloro-2-methoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 19) was prepared as in Scheme 2 using 4-chloro-2-iodo-1-methoxybenzene.

[0127] 1 1H NMR (400 MHz, CDCl3) δ 7.45 (d, J = 2.7 Hz, 1H), 7.23 - 7.04 (m, 5H), 6.75 (d, J = 8.7 Hz, 1H), 6.39 (d, J = 8.3 Hz, 1H), 5.40 - 5.34 (m, 1H), 4.54 - 4.42 (m, 1H), 3.82 (s, 3H), 3.32 - 3.05 (m, 6H), 2.90 (dd, J = 15.6, 7.5 Hz, 1H), 2.81 - 2.58 (m, 2H), 2.45 (dd, J = 15.6, 7.1 Hz, 1H), 2.38 - 2.30 (m, 2H), 2.30 - 2.15 (m, 2H), 2.08 - 1.94 (m, 4H). HRMS (ESI) m / z: [M + H] + Calculated value 443.2102; Measured value: 443.2087.

[0128] The following compounds were prepared as in representative Scheme 2 using non-commercial aryl iodides instead of Compound 4. The synthesis of this iodide is included below.

Chemical formula

[0129] N-((1R,2R)-1-(3-chloro-5-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(2,3-dihydro-1H-inden-2-yl)acetamide (Example 20) was prepared as in Scheme 2 using 1-chloro-3-cyclopropoxy-5-iodobenzene.

[0130] 11H NMR (400 MHz, CDCl3) δ 7.13 (m, 4H), 6.98 (m, 2H), 6.89 (m, 1H), 5.95 (d, J = 7.6 Hz, 1H), 5.00 (d, J = 2.8 Hz, 1H), 4.22 (m, 1H), 3.67 (m, 1H), 2.97 (dd, J = 15.6, 7.7 Hz, 1H), 2.92 - 2.82 (m, 3H), 2.82 - 2.56 (m, 5H), 2.50 (dd, J = 15.6, 6.8 Hz, 1H), 2.36 (dd, J = 15.5, 6.6 Hz, 1H), 2.32 - 2.13 (m, 2H), 1.78 (m, 4H), 0.74 (m, 4H). HRMS (ESI) m / z: [M + H] + Calculated value: 469.2258; Measured value: 469.2241.

Chemical Structure

[0131] 1-Chloro-3-cyclopropoxy-5-iodobenzene. To a solution of 3-chloro-5-iodophenol (1.2 g, 4.7 mmol) in N,N-dimethylacetamide (15 mL) were added cesium carbonate (6.40 g, 19.6 mmol) and bromocyclopropane (2.6 mL, 33 mmol). The reaction mixture was heated in a sealed tube at 150 °C overnight. After 24 h, an additional 1.5 mL of bromocyclopropane was added and the reaction mixture was stirred for 24 h. The reaction mixture was cooled to room temperature, poured into water, and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, filtered, and concentrated. The reaction mixture was purified by column chromatography eluting with 0 - 50% ethyl acetate in hexane. The title compound was isolated as a colorless transparent liquid (646 mg, 47%).

[0132] GCS inhibition GCS inhibitors are known. Some GCS inhibitors, such as eliglustat and miglustat, have sufficient activity to inhibit GCS activity and have thus been proposed as suitable for the treatment of diseases associated with the accumulation of glycolipids. Unfortunately, these compounds and / or their pharmacological profiles are not entirely satisfactory. For example, miglustat can cross the blood-brain barrier (BBB), but does not achieve levels above its IC 50 in the CNS, and much of its effect is either off-target or due to its potential activity as a chemical chaperone for beta-glucosylceramidase. Eliglustat is more potent than miglustat in inhibiting GCS, but cannot cross the BBB. Thus, diseases that require a therapeutic agent to cross the BBB cannot be treated. As a result, there is a continuing need to provide new compounds that effectively and selectively inhibit GCS and, in some embodiments, can cross the BBB. Compounds of structural formula (I) exhibit these beneficial properties.

[0133] To demonstrate the ability of the GCS inhibitors of the present invention to reduce the accumulation of glycolipids within lysosomes and cross the BBB, the compounds of the present invention were prepared and assayed. The compounds of structural formula (I) are also more potent GCS inhibitors intracellularly and show improved metabolic stability compared to previous GCS inhibitors.

[0134] The compounds were screened for inhibition of GCS and MDR1 substrate recognition in disrupted cell and whole cell assays. The compounds of structural formula (I) were found to inhibit GCS at low nanomolar concentrations with little or no apparent recognition by MDR1. Furthermore, three days of intraperitoneal administration of the compound to mice resulted in a significant dose-dependent decrease in the glucosylceramide content of the brain, an effect not seen in mice co-administered with eliglustat tartrate.

[0135] The compounds of formula (I) retain activity against GCS and eliminate substrate specificity for the MDR1 protein. As a result, novel compounds that inhibit GCS in both the brain and peripheral organs are provided.

[0136] Assay Materials N-((1R,2R)-1-(2,3-Dihydrobenzo-[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)octanamide (eliglustat tartrate) was provided by Genzyme Corporation. 3 H]Vinblastine and 14 C]Mannitol were purchased from American Radiolabeled Chemicals (St Louis, MO).

[0137] GCS Inhibition in MDCKII Cells Parental (wild-type-) 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 two months.

[0138] Stock solutions (100 mM) of water-insoluble sphingolipid inhibitors were prepared by dissolving each inhibitor in 100% ethanol as described above (3). Next, the inhibitor-ethanol solution was diluted 50-fold with 2 mM defatted bovine serum albumin-phosphate buffered saline to create a water-soluble sphingolipid inhibitor-bovine serum albumin complex. The inhibitor-bovine serum albumin complex was sterile filtered and stored at -20 °C. Prior to use, a portion of the inhibitor-bovine serum albumin complex was further diluted with Opti-F12 to create the treatment solution. Equal amounts of bovine serum albumin and ethanol were added to the control cultures. Cells (5×10 5Cells were seeded into a 10-cm culture dish containing 10 ml of Opti-F12 with 5% FBS. 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.

[0139] Cell lipid analysis After inhibitor treatment, total cell lipids of wild-type MDCKII cells were extracted as described in detail above (10). Briefly, cells were washed with ice-cold phosphate-buffered saline, fixed with methanol, and collected with a rubber spatula. Next, chloroform was added to form a single phase with a theoretical ratio of chloroform:methanol:water of 1:2:0.8 (v / v / v). Cell debris and proteins were removed by centrifugation at 2200 x g for 30 minutes. Chloroform and 0.9% NaCl were added to separate the supernatant. The lower organic phase containing neutral sphingoglycolipids and lipids was washed with methanol and 0.9% NaCl and subjected to base hydrolysis and acid hydrolysis (10). A portion of the purified sphingoglycolipids normalized to 100 nmol of total phospholipids was analyzed by high-performance thin-layer chromatography. The thin-layer chromatography separation was performed twice. Plates pretreated with 1% sodium borate were first developed in a solvent system consisting of chloroform / methanol (98 / 2, v / v). After air-drying, the plates were then developed in a solvent system containing chloroform / methanol / water (70 / 30 / 4, v / v / v). The level of glucosylceramide was detected by charring with 8% copper sulfate in 8% phosphoric acid and quantified by densitometric scanning using ImageJ, NIH Image. The image data were analyzed, and the IC 50 of each inhibitor was calculated using GraphPad Prism (version 5.03).

[0140] Reduction of glucosylceramide and glucosylsphingosine in the brain of CBE mice Reduction of the brain's glucosylceramide and glucosylsphingosine content by an inhibitor of GCS was determined by using mice with the Gba1 mutant allele called D409V / null. These are knock-in mice with a Gba1 point mutation encoding valine (V) at position 409 of the wild-type (WT) aspartic acid (D) of one missense allele and a null heterozygous allele [D409V / null (9V / null)]. D409V / null and wild-type littermates were sex-mixed and had a mixed C57BL / 129Sv / FVB genetic background but were otherwise identical. (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.) Mice were treated from day 15 after parturition with intraperitoneal conduritol B epoxide (CBE), an irreversible inhibitor of beta-glucocerebrosidase (25 mg / kg / day) in all groups. Mice were co-treated for 14 days with a GCS inhibitor or vehicle control. Mice were sacrificed 2 hours after the last injection, and tissues including the brain, liver, spleen, kidney, and lung were analyzed for glucosylceramide and glucosylsphingosine by mass spectrometry.

[0141] Mouse Tissue Lipid Analysis Lipid extraction from the liver, kidneys, and brain was performed as described above (7). Briefly, frozen liver (about 0.5 g), two kidneys (about 0.3 g), and whole brain (about 0.4 g) were individually homogenized with a Tri-R homogenizer in sucrose buffer (250 mM sucrose, pH 7.4, 10 mM HEPES, and 1 mM EDTA) at 0.2 g tissue / 1 mL of sucrose buffer. Each 0.8 mL of homogenate was mixed with 2 mL of methanol and 1 mL of chloroform, sonicated in a bath for 1 minute, and incubated at room temperature for 1 hour. Tissue debris was removed by centrifugation at 2,400 x 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 x 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 x gravity for 5 minutes, the lower layer was washed with 3 mL of methanol and 2.4 mL of 0.9% NaCl. The 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 x gravity for 5 minutes. The resulting lower phase was collected and dried under a stream of N2 gas.

[0142] Analysis of neutral sphingoglycolipids from mouse liver, kidney, and brain was processed after alkaline methanolysis. Kidney lipids were incubated with 2 mL of chloroform and 1 mL of 0.21 N NaOH in methanol for 2 h (kidney) or 7.5 h (liver and brain) at room temperature. Lipid extracts were normalized to 0.5 μmol of total phospholipid phosphate (liver and kidney) or 2 μmol of total phospholipid phosphate (brain) for high-speed thin-layer chromatography analysis. After alkaline methanolysis, brain lipids were passed through a silica gel column (7). Thin-layer chromatography plates impregnated with borate were developed in a 2-solvent system. The plates were first developed with chloroform / methanol (98:2, v / v). Next, plates loaded with kidney and liver lipids were developed with 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.

[0143] Assay results The activities of the compounds of structural formula (I) in the inhibition of GlcCer production in the cell disruption assay are summarized in Table 1.

[0144] [Table 3]

[0145] The following is the structure of the control compound used in all tests and assays disclosed herein. [Chemical formula]

[0146] In the present invention, it has been discovered that the chloro substituent at the 3-position of the phenyl ring provides an unexpected effect on the compound of structural formula (I) with respect to the MDCK IC50 inhibitor. As shown in Table 2 below, compounds of structural formula (I) having a chloro substituent at the 3-position of the phenyl ring were compared with the same compounds lacking the 3-chloro substituent. Typically, at least a 10-fold improvement was demonstrated.

[0147] As shown by the MLM T1 / 2 in Table 2, it has also been discovered that the chloro substituent at the 3-position generally provides an improvement in metabolic stability, as well as an improvement in drug exposure in the brain (Examples 5 and 7 in Table 2).

[0148] [Table 4]

[0149] Methods and Compositions The present invention provides a GCS inhibitor exemplified by a compound of structural formula (I) for the treatment of various diseases and conditions in which inhibition of 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 GCS provides a benefit, the method comprising administering to the individual in need thereof a therapeutically effective amount of a compound of structural formula (I).

[0150] Accordingly, the compounds of structural formula (I) can be used to treat a variety of diseases and conditions in which inhibition of GCS provides a benefit. Examples of such diseases and conditions include Tay-Sachs disease, types I, II, and III Gaucher disease, Sandhoff disease, and Fabry disease, Parkinson's disease (J.R. 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 levels, elevated glycated hemoglobin levels, lupus, and glomerular diseases selected from the group consisting of mesangial proliferative glomerulonephritis, focal segmental glomerulosclerosis, proliferative lupus nephritis, crescentic glomerulonephritis, and membranous nephropathy, but are not limited thereto.

[0151] Using the compound of structural formula (I) to treat disorders involving cell growth and division, including cancer, collagen vascular diseases, atherosclerosis, and renal hypertrophy in diabetic patients (see U.S. Pat. Nos. 6,916,802 and 5,849,326, each incorporated herein by reference), inhibiting the proliferation of arterial epithelial cells (see U.S. Pat. Nos. 6,916,802 and 5,849,326, each incorporated herein by reference), treating patients suffering from infectious diseases (M. Svensson et al., Infect. And Immun., 62:4404 - 4410 (1994)), preventing the host, i.e., the patient, from generating antibodies against a tumor (J. Inokuchi et al., Cancer Lett., 38:23 - 30 (1987)), and treating tumors (S. Hakomori Cancer Cells 3:461 - 470 (1991); J. Inokuchi et al., Cancer Res., 50:6731 - 6737 (1990); and (M. Ziche et al., Lab Invest., 67:711 - 715 (1992)) are also possible. Using the compound of structural formula (I), it is further possible to treat polycystic kidney disease, including both autosomal dominant and recessive forms (T.A. Natoli et al., Nat. Med. 16:788 - 792 (2010)).

[0152] The methods of the present invention can be achieved by administering the compound of structural formula (I) as an undiluted compound or as a pharmaceutical composition. Administration of the pharmaceutical composition, or the undiluted compound of structural formula (I), can be carried out 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 cause adverse reactions when administered. Further, a kit is provided that includes the compound of structural formula (I) and, optionally, a second therapeutic agent useful for the treatment of diseases and conditions in which inhibition of GCS provides a benefit, which is packaged separately or together, and a package insert describing the use of these active agents.

[0153] In many embodiments, the compound of structural formula (I) is administered with a second therapeutic agent useful for the treatment of a disease or condition in which inhibition of the GCS provides a benefit. The second therapeutic agent is different from the compound of structural formula (I). The compound of structural formula (I) and the second therapeutic agent can be administered simultaneously or sequentially to achieve the desired effect. Additionally, the compound of structural formula (I) and the second therapeutic agent can be administered from a single composition or two separate compositions.

[0154] The second therapeutic agent is administered in an amount that provides its desired therapeutic effect. The effective dosage range for each second therapeutic agent is known in the art, and the second therapeutic agent is administered to an individual in need thereof within such established range.

[0155] The compound of structural formula (I) and the second therapeutic agent can be administered together as a single unit dose or separately as multiple unit doses, and the compound of structural formula (I) can be administered before or after the second therapeutic agent. 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 with one or more second therapeutic agents, including, but not limited to, for example, enzyme replacement therapy, gene therapy, and isophagamine.

[0156] In a method of treating type 2 diabetes, the second therapeutic agent can be one or more of insulin (e.g., NOVOLIN®, NOVOLOG®, VELOSULIN®), sulfonylurea agents (e.g., DIABINESE®, GLUCOTROL®, GLUCOTROL XL®, DIABETA®, AMARYL®, ORINASE®, TOLINASE®, MICRONASE®, and GLYNASE®), metformin, α-glucosidase inhibitors (e.g., GLYSET®), thiazolidinediones (e.g., ACTOS® and AVANDIA®), nateglinide (STARLIX®), repaglinide (PRANDIN®), and combination drugs such as AVANDAMET® (AVANDIA® and metformin).

[0157] In a method of treating Parkinson's disease, the second therapeutic agent can be one or more of carbidopa / levodopa therapy, dopamine agonists (apomorphine hydrochloride, bromocriptine, rotigotine, pramipexole, ropinirole, pergolide), anticholinergic agents (benztropine mesylate, trihexyphenidyl hydrochloride, procyclidine), MAO-B inhibitors (selegiline, rasagiline), COMT inhibitors (entacapone, tolcapone), and other drugs including over-the-counter therapeutic agents (amantadine, rivastigmine tartrate, creatine, coenzyme Q10).

[0158] Diseases and conditions that can be treated by the present invention include, for example, Gaucher disease, Fabry disease, Tay-Sachs disease, and diabetes. In particular, type II and type III Gaucher diseases can be treated because the compound of structural formula (I) can cross the BBB. Conventional GCS inhibitors have not been able to treat various diseases associated with the accumulation of glycolipids because they cannot cross the BBB or have low efficacy and selectivity.

[0159] In this method, one or more compounds of structural formula (I), typically formulated according to pharmaceutical practice, are administered to a human in need thereof. Whether such treatment is indicated depends on the individual case and is subject to a medical evaluation (diagnosis) taking into account the existing signs, symptoms, and / or dysfunctions, the risk of developing specific signs, symptoms, and / or dysfunctions, and other factors.

[0160] The compounds of structural formula (I) can be administered by any suitable route, for example, orally, buccally, by inhalation, sublingually, rectally, intravaginally, intracisternal or intrathecal by lumbar puncture, transurethrally, intranasally, percutaneously, i.e., transdermally, or parenterally (including intravenous, intramuscular, subcutaneous, intracoronary, intradermal, intramammary, intraperitoneal, intra-articular, intrathecal, retrobulbar, intralung injection, and / or surgical implantation at a specific site). Parenteral administration can be achieved using needles and syringes or using high-pressure techniques.

[0161] Pharmaceutical compositions include those in which the compounds of structural formula (I) are administered in an amount effective to achieve their intended purpose. The exact formulation, route of administration, and dosage will be determined by the individual physician in view of the diagnosed condition or disease. Dosage and dosing intervals can be adjusted individually to provide a level of the compound of structural formula (I) that is sufficient to maintain a therapeutic effect.

[0162] The toxicity and therapeutic efficacy of the 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, which is 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., suppression of tumor growth) is the therapeutic index. The dosage can vary within this range depending on the dosage form used and the route of administration utilized. Determination of a therapeutically effective amount is well within the capabilities of a person skilled in the art, especially in view of the detailed disclosure provided herein.

[0163] The therapeutically effective amount of the compound of formula (I) required for use in therapy will vary depending on the nature of the condition being treated, the length of time for which activity is desired, as well as the age and condition of the patient and will ultimately be determined by the attending physician. The dosage and interval can be adjusted individually to provide a plasma level of the compound of formula (I) sufficient to maintain the desired therapeutic effect. The desired dosage can be conveniently administered as a single dose or as multiple doses in divided doses, for example, once, twice, three, or four or more times daily at appropriate intervals. Multiple doses are often desirable or necessary. For example, the compound of formula (I) can be delivered as 4 doses (q4d×4) delivered as 1 dose per day at 4-day intervals, 4 doses (q3d×4) delivered as 1 dose per day at 3-day intervals, 1 dose per day delivered at 5-day intervals (qdx5), 1 dose per week for 3 weeks (qwk3), 5 daily doses, followed by a 2-day break, followed by 5 more daily doses (5 / 2 / 5), or in any dosing regimen determined to be appropriate for the situation.

[0164] The compound of formula (I) used in the method of the present invention can 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. For example, the compound of formula (I) can be administered in an amount 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 dosages from 0.005 to 500 milligrams per dose.

[0165] The dosage of the composition containing the GCS inhibitor of structural formula (I) or the composition containing the same may be from about 1 ng / kg to about 200 mg / kg, from about 1 μg / kg to about 100 mg / kg, or from 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 any dosage including, but not limited to, 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, 350 μg / kg, 375 μg / kg, 400 μg / kg, 425 μg / kg, 450 μg / kg, 475 μg / kg, 500 μg / kg, 525 μg / kg, 550 μg / kg, 575 μg / kg, 600 μg / kg, 625 μg / kg, 650 μg / kg, 675 μg / kg, 700 μg / kg, 725 μg / 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. The foregoing dosages are exemplary of average cases, but there may be individual cases that merit higher or lower dosages, and such cases are within the scope of the present invention. In practice, a physician determines the actual dosage regimen that is most suitable for an individual patient, which may vary depending on the age, weight, and response of the particular patient.

[0166] The compounds of the present 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 present invention are formulated in conventional manner using one or more physiologically acceptable carriers including excipients which facilitate processing of the compounds of formula (I) and auxiliary agents.

[0167] These pharmaceutical compositions can be produced, for example, by conventional processes of mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating, entrapping, or lyophilizing. Suitable formulations depend on the selected route of administration. When a therapeutically effective amount of the compound of formula (I) is administered orally, the composition is typically in the form of tablets, capsules, powders, solutions, or elixirs. When administered in tablet form, the composition can further contain a solid carrier such as gelatin or an adjuvant. Tablets, capsules, and powders contain from about 0.01% to about 95%, preferably from about 1% to about 50%, of the compound of formula (I). When administered in liquid form, a liquid carrier such as water, petroleum, or animal or vegetable oils can be added. The liquid form of the composition can further contain a physiological saline solution, a dextrose or other saccharide solution, or a glycol. When administered in liquid form, the composition contains from about 0.1% by weight to about 90% by weight, preferably from about 1% by weight to about 50% by weight, of the compound of formula (I).

[0168] When a therapeutically effective amount of the compound of formula (I) is administered by intravenous, cutaneous, or subcutaneous injection, the composition is in the form of a pyrogen-free parenterally acceptable aqueous solution. Taking into account pH, isotonicity, stability, etc., the preparation of such parenterally acceptable solutions is within the skill of the art. Compositions preferred for intravenous, cutaneous, or subcutaneous injection typically contain an isotonic vehicle.

[0169] The compounds of formula (I) can be readily combined with pharmaceutically acceptable carriers well-known in the art. For oral administration to a patient to be treated, such carriers enable the active agent to be formulated as tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions and the like. Pharmaceutical preparations for oral use can be obtained by adding the compound of formula (I) to a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries if desired, to obtain a tablet or dragee core. Suitable excipients include, for example, fillers and cellulose preparations. Disintegrating agents can be added if desired.

[0170] The compounds of formula (I) can be formulated for parenteral administration by injection, for example, bolus injection or continuous infusion. Preparations for injection can be provided in unit dosage form in, for example, ampoules or multi-dose containers with a preservative added. The composition can take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending agents, stabilizers, and / or dispersing agents.

[0171] Pharmaceutical compositions for parenteral administration contain an aqueous solution of the active agent in water-soluble form. In addition, suspensions of the compounds of formula (I) can be prepared as suitable 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 to enable the preparation of highly concentrated solutions. Alternatively, the composition can be in powder form for constitution with a suitable vehicle, for example, sterile water free from pyrogens, before use.

[0172] The compounds of formula (I) can also be formulated, for example, into rectal compositions such as suppositories or retention enemas, which contain conventional suppository bases. In addition to the previously described formulations, the compounds of formula (I) can also be formulated as depot preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds of formula (I) can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or with ion exchange resins.

[0173] In particular, the compounds of formula (I) can be administered orally, buccally, or sublingually, in the form of tablets containing excipients such as starch or lactose, or in capsules or ovules, either alone or in admixture with excipients, or in the form of elixirs or suspensions containing flavoring or coloring agents. Such liquid preparations can be prepared using pharmaceutically acceptable additives such as suspending agents. The compounds of formula (I) can also be administered parenterally, for example, by intravenous, intramuscular, subcutaneous, or intracoronary injection. For parenteral administration, the GCS inhibitor is optimally used in the form of a sterile aqueous solution which may contain other substances, such as salts, or monosaccharides such as mannitol or glucose, to render the solution isotonic with the blood.

[0174] As an additional embodiment, the present invention includes a kit comprising one or more compounds or compositions packaged in a manner that facilitates the use of one or more compounds or compositions for practicing the methods of the present invention. In one simple embodiment, the kit comprises a compound or composition described herein as being useful for practicing the method (e.g., a composition comprising a compound of structural formula (I) and an optional second therapeutic agent), packaged in a container such as a sealed bottle or sealed container, and a label describing the use of the compound or composition for practicing the methods of the present invention is affixed to or included in the kit. Preferably, the compound or composition is packaged in unit dosage form. The kit can further comprise a device suitable for administering the composition according to the intended route of administration.

[0175] Previous GCS inhibitors have had properties that have hindered their development as therapeutic agents. In accordance with an important feature of the present invention, the compounds of structural formula (I) have been synthesized and evaluated as inhibitors of GCS, particularly as inhibitors having the ability to cross the BBB. Current GCS inhibitors are characterized by inhibition of GCS at low nanomolar concentrations, high specificity, and lack of β-glucosylceramidase binding.

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Claims

1. A compound having the following structural formula (I), 【Chemical 1】 wherein R 1 and R 2 are independently H, C 1~4 alkyl, OC 1~4 alkyl, C 3~6 cycloalkyl, or OC 3~6 cycloalkyl, each optionally substituted with one or more F, and one carbon of the cycloalkyl group in said C 3~6 cycloalkyl or said OC 3~6 cycloalkyl is optionally substituted by O, or R 1 and R 2 together have one or two carbon atoms substituted by CH 3 and form an OCH 2 CH 2 O ring NR 5 R 5’ is selected from the group consisting of 1-pyrrolidinyl, 3-fluoro-1-pyrrolidinyl, and 1-azetidinyl, R 3 is H or F, and G is CH 2 or O, and R 4 is H or CH 3 and R 6 is H, C 1~3 alkyl, C 3~6 cycloalkyl, or CD 3 and R 7 and R 8 are each independently H, F, or OC 1~3 alkyl, a compound, or a pharmaceutically acceptable salt thereof.

2. R 1 is -H, -OC(CH 3 ) 3 , -OCH 3 , -OCF 3 , -OCHF 2 , or a group of the following formula: 【Chemical Formula 2】 The compound according to claim 1, wherein

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

4. R 2 The compound according to any one of claims 1 to 3, wherein R is H.

5. R 1 and R 2 together form the basis of the following formula: [Chemical Formula 3] The compound according to claim 1, which forms

6. R 3 The compound according to any one of claims 1 to 5, wherein R is H or F.

7. R 4 is H or CH 3 The compound according to any one of claims 1 to 6.

8. -NR 5 R 5’ is based on the following formula: [Chemical Formula 4] The compound according to any one of claims 1 to 7, wherein

9. R 1 , R 2 , R 7 , and R 8 wherein each of which is -H, the compound according to any one of claims 1 to 8.

10. R 7 is H, and R 8 is OCH 3 or R 7 is OCH 3 or R 8 is H, or R 7 is H, R 8 is H, the compound according to any one of claims 1 to 9.

11. The partial structure in the structural formula (I): [Chemical Formula 5] is a group consisting of the following groups: 【Chemical Formula 6】 The compound according to any one of claims 1 to 10, which is selected from the group consisting of

12. A compound of the following formula: 【Chemical Formula 7】 【Chemical Formula 8】 The compound selected from the group consisting of

13. A pharmaceutical composition comprising the compound according to any one of claims 1 to 12, which is used for treating a disease or condition in which inhibition of GCS provides a benefit, and the disease or condition in which inhibition of GCS provides a benefit is Gaucher disease, Fabry disease, Sandhoff disease, Tay-Sachs disease, Parkinson's disease, multiple myeloma, ADPKD, type 2 diabetes, hypertrophy or hyperplasia associated with diabetic nephropathy, an increase in plasma TNF-α level, an increase in blood glucose level, an increase in glycated hemoglobin level, glomerular disease, or lupus, a pharmaceutical composition.

14. The pharmaceutical composition according to claim 13, wherein the Gaucher disease is type I, type II, or type III Gaucher disease.

15. The pharmaceutical composition according to claim 13, wherein the glomerular disease is selected from the group consisting of mesangial proliferative glomerulonephritis, focal segmental glomerulosclerosis, proliferative lupus nephritis, crescentic glomerulonephritis, and membranous nephropathy.

16. The pharmaceutical composition according to claim 13, wherein the disease or condition in which inhibition of GCS provides a benefit is selected from the group consisting of disorders associated with cell proliferation, disorders associated with cell division, collagen vascular diseases, atherosclerosis, renal hypertrophy in diabetic patients, proliferation of arterial epithelial cells, infectious diseases, tumors, and polycystic kidney disease.

17. A pharmaceutical composition used for inhibiting glucosylceramide synthase or reducing the concentration of sphingolipids, the pharmaceutical composition comprising the compound according to any one of claims 1 to 12.

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