Methods for treating neurological symptoms associated with lysosomal storage diseases

Abstract

To provide a pharmaceutical composition effective in alleviating or managing neurological symptoms associated with Gaucher disease type 3.SOLUTION: There is provided a pharmaceutical composition for use in a method of enhancing neuronal connectivity in the brain of a subject, which comprises an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or prodrug thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention particularly relates to methods for treating or preventing symptoms and disorders associated with, for example, lysosomal storage diseases, using a quinuclidine compound of formula (I), optionally in combination with an enzyme replacement therapy drug. These methods enhance neuronal connectivity in the brain of a subject, increase brain tissue volume, or prevent or delay the loss of brain tissue volume in a subject. The present invention also relates to methods for monitoring the progression or regression of, or assessing the onset of, neurological disorders associated with lysosomal storage diseases, in which the brain tissue volume of a subject is measured. [Background technology]

[0002] Lysosomal storage disorders Lysosomal storage diseases (LSDs) are a group of approximately 50 rare, inherited metabolic disorders caused by defects in lysosomal function. Patients with LSDs typically accumulate harmful levels of substrate (i.e., storage material) in lysosomes due to a deficiency or defect in an enzyme responsible for substrate metabolism or due to a deficiency in an enzyme activator essential for proper enzyme function. Most LSDs are caused by a defect or deficiency in a single enzyme, usually an enzyme involved in lipid or glycoprotein metabolism. Some of the more common LSDs include Gaucher disease, Fabry disease, and Niemann-Pick disease (type C). Gaucher, Fabry, and Niemann-Pick are examples of sphingolipidosis. Each of these diseases is associated with a set of symptoms caused directly or indirectly by an underlying genetic defect. Consequently, it is often difficult to predict whether the symptoms or disorders associated with each disease can be effectively treated with different treatment methods. Symptoms common to several LSDs include alterations in saccadic eye movements, cognitive impairment, and gait disorders, such as ataxia. These symptoms are particularly common in Gaucher disease (eg, type 3) and Niemann-Pick disease (type C).

[0003] Gaucher disease (GD) is a rare autosomal recessive lysosomal storage disorder. Patients with GD have mutations in the GBA1 gene, which encodes glucosylceramidase (GC), also known as beta-glucocerebrosidase. This enzyme is responsible for breaking down glycosphingolipids into their constituent components, such as glucosylceramide (GLC; also known as glucocerebroside) into glucose and ceramide. Monocytes and macrophages have particularly high levels of lysosomes containing GLC, and in GD patients, these cells become enlarged and accumulate toxic concentrations of GLC. These so-called "Gaucher cells" accumulate in several organs, including bone, bone marrow, spleen, liver, lungs, and brain. This leads to systemic splenomegaly, hepatomegaly, anemia, thrombocytopenia, leukopenia, osteopenia, osteonecrosis, and other pathological abnormalities.

[0004] There are three subtypes of Gaucher disease, which differ in age of onset, severity, and the presence of neurological findings. Type 1 Gaucher disease (GD-1), a non-neuronopathic form of Gaucher disease, is the most common form, with a median age of diagnosis of 28 years and a slightly shorter life expectancy. In GD-1, the GC enzyme retains some function and there is no neurological involvement. Type 2 Gaucher disease (GD-2) is an acute neuropathic form of Gaucher disease, diagnosed in infancy, with severe neurological involvement, and death usually within the first two years of life. The GC enzyme in type 2 patients is significantly impaired in function compared with GD-1. Type 3 Gaucher disease (GD-3) is a chronic neuropathic form of Gaucher disease, diagnosed in childhood, with a slow worsening of neurological involvement, and a life expectancy usually within 30 years. Symptoms of GD-3 include splenic and liver abnormalities, fatigue, bleeding, seizures, and supranuclear gaze palsy. Neurological findings in GD-3 patients vary over the course of the disease. The disease progresses gradually with the progression of GD-3. One of the more debilitating features is gaze paresis, which is a defect in the neural pathways that control saccadic eye movements. During the early phase of the disease, horizontal saccades are slowed. The disease progresses to complete horizontal saccade paresis and varying degrees of vertical saccade paresis. The VOR may also be impaired in GD-3 patients. These features of the disease can severely affect the quality of life of GD-3 patients and interfere with their educational and employment prospects.

[0005] Existing treatments for GD-1 and GD-3 are limited to recombinant enzyme replacement therapy (ERT) using imiglucerase, velaglucerase, or taliglucerase, and substrate reduction therapy (SRT) using miglustat or eliglustat. See, for example, Non-Patent Document 1. The primary treatment regimen, imiglucerase, is a recombinant version of human GC, produced from Chinese hamster ovary cells and administered slowly (typically over 1–2 hours) by intravenous infusion every 1–2 weeks. Velaglucerase, another recombinant human GC analogue, has been available in the United States since 1998 and is produced from a fibrosarcoma cell line and approved by the FDA in 2010. Taliglucerase is similar, produced using genetically modified carrot root cells and approved since 2012. All of these treatments require IV administration in a hospital or other medical facility, and the recombinant enzymes do not cross the blood-brain barrier, resulting in failure to treat the neurological symptoms of GD. Thus, these ERT regimens have proven effective in treating GD-1 patients, but in GD-3 patients, they are only effective in treating non-neurological symptoms of the disease.

[0006] Substrate reduction therapy is an alternative approach to treating GD. The goal of this therapy is to reduce GLC accumulation by inhibiting the enzyme responsible for GLC synthesis. Glucosylceramide synthase (GCS), also known as UDP-glucoseceramide synthase, is the enzyme that catalyzes the first glycosylation step of ceramide to form glucosylceramide.

[0007] GCS inhibitors have been proposed for treating various diseases, including glycolipid storage diseases and lysosomal storage diseases, including Gaucher disease. See, for example, U.S. Patent No. 5,949,999 (Actelion Pharm. Ltd.). Miglustat (Zavesca) is an imino-glucose GCS inhibitor. It is an N-alkylated iminosugar that acts as a reversible competitive inhibitor of GCS, binding to the active site of the enzyme. While it has been developed to treat neurotoxic forms of GD, GD-2, and GD-3, it is only approved by the FDA as second-line therapy for the treatment of patients with mild to moderate GD-1 (patients who should not be able to receive ERT treatment). Miglustat crosses the blood-brain barrier, but clinical trials have shown no efficacy in treating the neurological manifestations of GD-3. Eliglustat is also a GCS inhibitor and is a ceramide analog. It is FDA-approved only for the treatment of systemic symptoms in patients with GD-1.

[0008] Niemann-Pick disease type C (NPC) is also a lysosomal storage disease. Its cause differs significantly from Gaucher disease in some respects, but the end result is similar. NPC is caused by mutations in either the NPC1 or NPC2 gene. NPC1 is a membrane protein that mediates intracellular transport of cholesterol to post-lysosomal destinations. In particular, NPC1 acts in concert with NPC2 to facilitate the export of cholesterol from endosomal / lysosomal compartments. Unesterified cholesterol released from low-density lipoproteins in the lumen of late endosomes / lysosomes is transferred by NPC2 to the cholesterol-binding pocket of NPC1. Approximately 95% of NPC patients have mutations in NPC1, while the majority of the remainder have mutations in NPC2. One of the consequences of this disrupted cholesterol transport is the loss of cholesterol in the liver, spleen, and liver. The accumulation of cholesterol and glycosphingolipids (including GLC) in brain cells is a hallmark of NPC, such as GD-3, and is characterized by the gradual onset of supranuclear gaze paresis, including horizontal and vertical saccade paresis.

[0009] Another group of diseases and disorders commonly associated with saccadic gaze paresis are the GM2-gangliosidoses (e.g., Tay-Sachs disease, Sandhoff disease, and AB variant GM2 gangliosidoses).

[0010] GM2 gangliosidosis, like Gaucher disease, is a lysosomal storage disease characterized by a genetic defect in glycosphingolipid metabolism. GM2 gangliosidosis is characterized by defects in the enzyme hexosaminidase A and / or its cofactor, GM2 activator protein, responsible for the degradation of GM2 to GM3. GM2 and GM3 are related gangliosides and are part of the same metabolic pathway in which glucosylceramide is degraded to ceramide. Therefore, GM3 is produced through a stepwise process beginning with the conversion of ceramide to glucosylceramide (by GLC), followed by conversion to galactosyl-glucosylceramide, then to GM3 (N-acetyl-α-neuramidinyl-galactosyl-glucosylceramide), and then to GM2 (N-acetyl-galactosyl-N-acetyl-α-neuramidinyl-galactosyl-glucosylceramide). Therefore, the pathological accumulation of GM2, which is characteristic of GM2 gangliosidosis, can be reversed by GCS inhibitors that block the initial synthesis step of glucosylceramide.

[0011] The quinuclidine compounds described herein have the activity of enzyme glucosylceramide synthase (GCS) inhibitor.These compounds are disclosed as being generally useful in treating lysosomal storage diseases, such as Fabry disease, Gaucher disease and Niemann-Pick disease.See, for example, Patent Document 2 and Patent Document 3. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] WO2005 / 068426 [Patent Document 2] WO2012 / 129084 [Patent Document 3] US2016 / 0361301 [Non-patent literature]

[0013] [Non-Patent Document 1] Lunawati L.Bennett&Chris Fellner,Pharmacotherapy of Gaucher Disease:Current and Future Options,P&T43(5):274-280,309(2018) Summary of the Invention [Problem to be solved by the invention]

[0014] There is a real need in the art to develop treatments that are effective in reducing or managing the neurological symptoms associated with Gaucher disease type 3. [Means for solving the problem]

[0015] The present invention relates to a quinuclidine compound (Compound 1) according to formula (I) or a pharmaceutically acceptable salt or prodrug thereof. [ka] (In the formula: R 1 is hydrogen, halogen (e.g., fluorine), cyano, nitro, hydroxy, thio, amino, C 1~6 - alkyl (e.g., methyl or ethyl), C 2~6 -Alkenyl, C 2~6 -alkynyl, C 1~6 -Alkyloxy, C 2~6-alkenyloxy, and C 2~6 -alkynyloxy, wherein said alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy, or alkynyloxy is optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from halogen, cyano, nitro, hydroxy, thio, and amino; R 2 and R 3 is C optionally substituted by one or more (e.g., 1, 2, or 3) halogens; 1~3 -alkyl, or R 2 and R 3 together form a cyclopropyl or cyclobutyl group optionally substituted with one or more (e.g., one or two) halogens; R 4 , R 5 , and R 6 is hydrogen, halogen, nitro, hydroxy, thio, amino, C 1~6 -alkyl, and C 1-6 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen, hydroxy, cyano, and C 1~6 -optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; A is halogen, hydroxy, thio, amino, nitro, C 1~6 Alkoxy, and C 1~6 alkyl).

[0016] In a first aspect, the present application provides a method for treating or preventing cognitive impairment and / or gait abnormalities, including ataxia, associated with a lysosomal storage disease in a subject, such as a subject in need thereof, the method comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to Formula (I). In other aspects, the present application further provides use of a quinuclidine compound described herein for treating or preventing cognitive impairment and / or gait abnormalities, including ataxia, associated with a lysosomal storage disease and / or for the manufacture of a medicament for treating or preventing cognitive impairment and / or gait abnormalities, including ataxia, associated with a lysosomal storage disease.

[0017] In a second aspect, the present application provides a method for enhancing neuronal connectivity in the brain in a subject, such as a subject in need thereof, the method comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to Formula (I). In another aspect, the present application further provides the use of a quinuclidine compound described herein for enhancing neuronal connectivity in the brain of a subject and / or for the manufacture of a medicament for enhancing neuronal connectivity in the brain of a subject.

[0018] In a third aspect, the application provides a method for increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject, such as a subject in need thereof, said method comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to Formula (I). In other aspects, the application further provides a quinuclidine compound described herein for use in increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject in need thereof and / or for the manufacture of a medicament for increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject in need thereof.

[0019] In a fourth aspect, the present application provides a method for monitoring the progression or regression of a neurological disorder associated with a lysosomal storage disease in a subject, wherein the subject is undergoing treatment comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound of Formula (I); said method comprising measuring the subject's brain tissue volume over a period of time during the course of treatment, e.g., using volumetric magnetic resonance imaging (vMRI), and assessing the degree of change in brain tissue volume over said period.

[0020] In a fifth aspect, the present application provides a method for assessing the onset of a neurological disorder associated with a lysosomal storage disease in a subject at risk of developing the neurological disorder, the method comprising: a) measuring the subject's brain tissue volume (e.g., using vMRI) and comparing it with a reference standard to assess whether the brain tissue volume is lower than the reference standard; and b) identifying the onset of the neurological disorder if the brain tissue volume identified in step (a) is lower than the reference standard; and optionally further comprising: c) initiating treatment of the subject by administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound of Formula (I), or a pharmaceutically acceptable salt or prodrug thereof.

[0021] Additional features and advantages of the compounds, compositions and methods disclosed herein will become apparent from the following detailed description. DETAILED DESCRIPTION OF THE INVENTION

[0022] While specific embodiments of the present disclosure will now be described with reference to preparations and schemes, it should be understood that such embodiments are merely illustrative and merely a few of the many possible specific embodiments that can represent applications of the principles of the present disclosure. Various changes and modifications will be apparent to those skilled in the art given the benefit of this disclosure and are deemed to be within the spirit and scope of the present disclosure as further defined in the appended claims.

[0023] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this disclosure belongs.Although any method and material similar or equivalent to those described herein can be used to implement or test the present invention, exemplary methods, devices, and materials are described here.All technical and patent publications cited herein are incorporated herein by reference in their entirety.Nothing herein should be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention.

[0024] The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of tissue culture, immunology, molecular biology, microbiology, cell biology and recombinant DNA, which are within the skill of the art.

[0025] All numerical designations, including ranges, e.g., pH, temperature, time, concentration, molecular weight, are approximations that are varied (+) or (-) by increments of 0.1 or 1.0, as appropriate. It is understood, although not always explicitly stated, that all numerical designations are preceded by the term "about." It is also understood, although not always explicitly stated, that the reagents described herein are exemplary only, and that equivalents of such are known in the art.

[0026] As used herein, the term "optionally substituted" is meant to be equivalent to the phrase "unsubstituted or substituted by."

[0027] As used herein, the phrase "in a method of treating or preventing" (e.g., the phrase "in a method of treating or preventing pain") is meant to be equivalent to the phrase "in the treatment or prevention of" (e.g., the phrase "in the treatment or prevention of pain").

[0028] As used in the specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a cell" includes a plurality of cells, including mixtures thereof. Unless otherwise stated or obvious from the context, the term "or" is understood to be inclusive when used herein. The term "including" is used interchangeably herein to mean the phrase "including, but not limited to."

[0029] As used herein, the terms "comprising" or "comprises" are intended to mean that the compositions and methods include the recited elements, but do not exclude others. "Substantially consisting of," when defining compositions and methods, is intended to mean excluding any other elements that are essentially important to the combination for the described purpose. Thus, a composition consisting essentially of the elements defined herein would not exclude trace contaminants from isolation and purification methods, as well as pharmaceutically acceptable carriers, e.g., phosphate-buffered saline, preservatives, and the like. "Consisting of" is intended to mean excluding not only trace elements of other components, but also substantial method steps for administering the compositions of the invention or for producing the compositions or achieving an intended result. Embodiments defined by each of these transitional terms are within the scope of the present invention. The use of the term "comprising" herein is intended to encompass "substantially consisting of" and "consisting of."

[0030] The terms "subject," "individual," or "patient" are used interchangeably herein to refer to vertebrates, e.g., mammals. Mammals include, but are not limited to, mice, rats, rabbits, monkeys, cows, sheep, pigs, dogs, cats, farm animals, sport animals, pets, horses, primates, and humans. In one embodiment, mammals include horses, dogs, and cats. In some embodiments, the mammal is a human, e.g., a human suffering from a particular disease or disorder, e.g., Gaucher disease (e.g., GD-3) or Niemann-Pick disease type C.

[0031] "Administering" is defined herein as a means of providing a drug or a composition containing a drug to a subject in a manner that results in the drug being present inside the subject's body. Such administration may be by any route, including, but not limited to, oral, transdermal (e.g., vaginal, rectal, oral mucosal), by injection (e.g., subcutaneous, intravenous, parenteral, intraperitoneal, intra-CNS), or by inhalation (e.g., oral or nasal). Pharmaceutical Preparations is of course given in a form suitable for each administration route.

[0032] "Treating" or "treatment" of a disease generally includes (1) inhibiting the disease, i.e., arresting or reducing the onset of the disease or its clinical symptoms; and / or (2) palliating the disease, i.e., causing regression of the disease or its clinical symptoms.

[0033] As used herein, " treating " and " treatment " also refer to either the reversal of cognitive impairment and / or gait abnormality of disease, or the stabilization of such symptoms.This is because the diseases and disorders described herein are progressive disorders - if not treated, patient's condition will continue to deteriorate.For example, in the early stage of disease, patient may suffer from mild cognitive impairment and / or gait abnormality, but as disease progresses, patient may develop much more severe symptoms.Therefore, treatment encompasses both the delay (for example, stabilization) of this progressive deterioration and the reversal (for example, improvement) of this progressive deterioration.

[0034] "Preventing" or "preventing" a disease generally involves preventing the development of clinical symptoms of the disease in a patient who may be susceptible to the disease but who has not yet experienced or exhibited symptoms of the disease.

[0035] As used herein, " preventing " or " preventing " also encompasses preventing the development of cognitive impairment and / or gait abnormality in patients who are suspected of having or diagnosed with the diseases or disorders described herein.Because the diseases and disorders described herein are progressive disorders, various signs and symptoms may gradually appear as the disease progresses.Therefore, for example, a patient may be diagnosed with GD-3 or NPC before cognitive impairment and / or gait abnormality begins to develop.In such patients, the treatment methods described herein can be effective in preventing the development of cognitive impairment and / or gait abnormality.

[0036] The term "mild paralysis" is synonymous with "complete paralysis" and includes any degree of loss of motor function of one or more skeletal muscles. Therefore, as used herein, the term "mild paralysis" encompasses both complete mild paralysis, i.e., complete and complete paralysis, and partial mild paralysis, i.e., partial and complete paralysis. Complete mild paralysis means that a muscle or muscle group, such as an extraocular muscle, has lost the ability to contract. Thus, the affected eye or eyes cannot move. Partial mild paralysis may manifest as inhibition of movement, slowness of movement, or other defects in movement. These may include loss of range of motion. When applied to saccades, this may include inhibition of saccade initiation (e.g., in response to a stimulus), changes in saccade frequency, changes in saccade peak velocity, changes in saccade amplitude, changes in intersaccade latency, and / or loss of the ability to maintain or shift gaze. As used herein, in some embodiments, mild paralysis includes ophthalmoplegia and / or ophthalmoplegia. Thus, the term encompasses both weakness and complete paralysis of the extraocular muscles, including any one or more of the superior rectus, inferior rectus, medial rectus, lateral rectus, inferior oblique, and superior oblique muscles of the eye. Weakness and / or complete paralysis may involve one or more of horizontal, vertical, or rotational movements.

[0037] The term "suffering" when associated with the term "treatment" refers to a patient or individual who has been diagnosed with a disease. The term "suffering" when associated with the term "prevention" refers to a patient or individual who is susceptible to a disease. A patient may be a person who is susceptible to a disease due to a history of the disease in their family. It can also refer to someone who is "at risk of" having a disease due to the presence of a genetic mutation associated with the disease. A patient who is at risk for a disease has not yet developed all or some of the characteristic symptoms of the disease.

[0038] The term "increase" with respect to methods for increasing brain tissue volume refers to increasing the volume of at least one, and preferably multiple, individual brain tissue regions, typically involving an increase in total brain tissue volume (i.e., the total volume of brain tissue in the subject).

[0039] An "effective amount" or "therapeutically effective amount" is an amount sufficient to produce beneficial or desired results. An effective amount may be administered in one or more administrations, applications, or dosages. Such delivery depends on many variables, including the duration of use of individual dosage units, the bioavailability of the therapeutic agent, and the route of administration. However, it is understood that the specific dosage level of a therapeutic agent of the present invention for any particular subject will depend on various factors, including, for example, the activity of the specific compound used, the subject's age, weight, general health, sex, and diet, the timing of administration, the rate of excretion, the drug combination, and the severity and administration form of the particular disorder being treated. Treatment dosages may generally be titrated to optimize safety and efficacy. Typically, dosage-effect relationships from in vitro and / or in vivo studies can initially provide useful guidance regarding appropriate dosages to administer to patients. Generally, it will be desirable to administer an amount of the compound effective to achieve serum levels commensurate with concentrations found to be effective in vitro. Determination of these parameters is well within the skill of the art. These considerations, as well as effective formulations and administration procedures, are well known in the art and are described in standard textbooks. Retaining this definition, as used herein, the term "therapeutically effective amount" refers to an amount sufficient to treat (e.g., ameliorate) one or more symptoms associated with a disease or disorder described herein (e.g., in any of Methods 1 et seq.), ex vivo, in vitro, or in vivo.

[0040] As used herein, the term "pharmaceutically acceptable excipient" includes any of the standard pharmaceutical excipients, including carriers such as phosphate buffered saline solution, water, and emulsions, such as oil / water or water / oil emulsions, as well as various types of wetting agents. Pharmaceutical compositions also include stabilizers and preservatives. For example, examples of carriers, stabilizers, and adjuvants can be found in Remington's Pharmaceuticals, Inc. See Sciences (20th ed., Mack Publishing Co. 2000).

[0041] As used herein, the term "prodrug" refers to a pharmacological derivative of a parent drug molecule that requires spontaneous or enzymatic biotransformation in an organism to release the active drug. For example, a prodrug is a modified form or derivative of a quinuclidine compound described herein that has a group cleavable under certain metabolic conditions, which, when cleaved, becomes a quinuclidine compound described herein, e.g., a compound of Formula (I). Such prodrugs then become pharmaceutically active in vivo when they undergo solvolysis or enzymatic degradation under physiological conditions. As used herein, prodrug compounds may be referred to as single, double, triple, etc., depending on the number of biotransformation steps required to release the active drug in an organism and the number of functional groups present in the precursor form. Prodrug forms often offer advantages of solubility, tissue compatibility, or delayed release in mammalian organisms.

[0042] Commonly known prodrugs in the art include, for example, esters prepared by reaction of an acid compound with a suitable alcohol, or by reaction of an acid compound with an acylated base derivative. Examples of prodrug derivatives include well-known acid derivatives such as amides prepared by reaction with amines, which are basic groups that react to form prodrugs. Other prodrug derivatives may be combined with other features disclosed herein to enhance bioavailability. Thus, those skilled in the art will understand that, for example, some of the compounds disclosed herein that have free amino or hydroxy groups can be converted into prodrugs. Prodrugs include compounds having an amino acid residue or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues covalently attached via peptide bonds to free amino, hydroxy, or carboxylic acid groups of the compounds disclosed herein. Amino acid residues include the 20 naturally occurring amino acids, commonly represented by their three-letter symbols, including 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Prodrugs also include compounds having a carbonate, carbamate, amide, or alkyl ester moiety covalently bonded to any of the above substituents disclosed herein.

[0043] As used herein, the term "pharmaceutically acceptable salt" means a pharmaceutically acceptable acid addition salt or a pharmaceutically acceptable base addition salt of the presently disclosed compounds that may be administered without any substantial undesired biological effects or any adverse interactions as a result with other components of the pharmaceutical composition in which it may be included.

[0044] As used herein, "C 1~6 The term "-alkyl" means a saturated, straight-chain or branched free radical consisting essentially of 1 to 6 carbon atoms and a corresponding number of hydrogen atoms. 1-6 -Alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, and isobutyl. 1~6-alkyl groups will be readily apparent to those of ordinary skill in the art given the benefit of this disclosure. 1~3 -alkyl", "C 1~4 The terms "-alkyl" and the like have the same meaning, i.e., a saturated, linear or branched free radical consisting essentially of 1 to 3 (or 4) carbon atoms and the corresponding number of hydrogen atoms.

[0045] As used herein, "C 2~6 The term "-alkenyl" refers to an unsaturated, linear or branched free radical consisting essentially of 2 to 6 carbon atoms and a corresponding number of hydrogen atoms, and containing at least one carbon-carbon double bond. 2~6 -alkenyl groups include ethenyl, prop-1-enyl, prop-2-enyl, isopropenyl, but-1-enyl, 2-methyl-prop-1-enyl, and 2-methyl-prop-2-enyl. 2~6 -alkenyl groups will be readily apparent to those of skill in the art given the benefit of this disclosure.

[0046] As used herein, "C 2~6 The term "alkynyl" refers to an unsaturated, linear or branched free radical consisting essentially of 2 to 6 carbon atoms and a corresponding number of hydrogen atoms, and containing at least one carbon-carbon triple bond. 2~6 -alkynyl groups include ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, and 3-methyl-but-1-ynyl. 2~6 -alkynyl groups will be readily apparent to those of skill in the art given the benefit of this disclosure.

[0047] As used herein, "C 1~6 The term "-alkyloxy" means a saturated, linear or branched free radical consisting essentially of 1 to 6 carbon atoms (and the corresponding number of hydrogen atoms) and oxygen atoms. 1~6 -Alkyloxy group is an oxygen atom The bond is via an example C 1~6 -Alkyloxy groups include methyloxy, ethyloxy, n-propyloxy, isopropyloxy, n-butyloxy, and isobutyloxy. 1~6 -alkyloxy groups will be readily apparent to those of ordinary skill in the art given the benefit of this disclosure. 1~3 -alkyloxy", "C 1~4 The terms "-alkyloxy" and the like have the same meaning, i.e., a saturated linear or branched free radical consisting essentially of 1 to 3 (or 4) carbon atoms (and the corresponding number of hydrogen atoms) and oxygen atoms, in which the group is attached via an oxygen atom.

[0048] As used herein, "C 2~6 The term "-alkenyloxy" refers to an unsaturated, linear or branched free radical consisting essentially of 2 to 6 carbon atoms (and the corresponding number of hydrogen atoms) and oxygen atoms, and containing at least one carbon-carbon double bond. 2~6 The -alkenyloxy group is bonded via an oxygen atom. 2~6 -an alkenyloxy group is ethenyloxy; others will be readily apparent to those of skill in the art given the benefit of this disclosure.

[0049] As used herein, "C 2~6 The term "alkynyloxy" refers to an unsaturated, linear or branched free radical consisting essentially of 2 to 6 carbon atoms (and the corresponding number of hydrogen atoms) and oxygen atoms, and containing at least one carbon-carbon triple bond. 2~6 The -alkenyloxy group is bonded via an oxygen atom. 2~6 -an alkenyloxy group is ethynyloxy; others will be readily apparent to those of skill in the art given the benefit of this disclosure.

[0050] As used herein, the term "heteroaryl" refers to an aromatic free radical having 5 or 6 atoms (i.e., ring atoms) forming a ring, wherein 1 to 5 ring atoms are carbon and the remaining 1 to 5 ring atoms (i.e., hetero ring atoms) are independently selected from the group consisting of nitrogen, sulfur, and oxygen. Exemplary 5-membered heteroaryl groups include furyl, thienyl, thiazolyl (e.g., thiazol-2-yl), pyrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl, imidazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 6-membered heteroaryl groups include pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, 1,2,4-triazinyl, benzoxazolyl, benzothiazolyl, benzisothiazolyl, benzisoxazolyl, and benzimidazolyl. Other heteroaryl groups will be readily apparent to those of ordinary skill in the art given the benefit of this disclosure. In general, heteroaryl groups are typically attached to the main structure via a carbon atom. However, one skilled in the art will recognize that certain other atoms, for example, heterocyclic atoms, can be attached to the main structure.

[0051] As used herein, the term "aryl" refers to an aromatic free radical having 5 or 6 atoms forming a ring (i.e., ring atoms), all of which are carbon. An exemplary aryl group is a phenyl group.

[0052] As used herein, the term "aliphatic" refers to a non-aromatic compound containing carbon and hydrogen atoms, e.g., 1 to 9 carbon atoms. Aliphatic compounds may be linear or branched, may contain one or more ring structures, and may contain one or more carbon-carbon double bonds (provided that the compound does not contain an unsaturated ring structure with aromatic character). Examples of aliphatic compounds include ethane, propylene, cyclobutane, and cyclohexadiene.

[0053] As used herein, the terms "halo" and "halogen" refer to fluorine, "Halogen" refers to chlorine, bromine, or iodine. These terms are used interchangeably and may refer to a halogen free radical group or the halogen atom itself. Those skilled in the art will be able to easily ascertain this identity given the context in which the term is used in this disclosure.

[0054] As used herein, the term "cyano" refers to a free radical having a carbon atom linked to a nitrogen atom through a triple bond. The cyano radical is bonded through that carbon atom.

[0055] As used herein, the term "nitro" refers to the -NO2 radical attached through its nitrogen atom.

[0056] As used herein, the terms "hydroxy" and "hydroxyl" refer to an -OH radical attached through its oxygen atom. The term "thio" refers to an -SH radical attached through its sulfur atom.

[0057] As used herein, the term "amino" refers to a free radical having a nitrogen atom and one or two hydrogen atoms.Therefore, the term "amino" generally refers to primary and secondary amines.In this regard, as used herein, tertiary amines are represented by the general formula RR'N- (wherein R and R' are carbon radicals that may or may not be identical).Nevertheless, the term "amino" may be used generally herein to describe primary, secondary, or tertiary amines, and those skilled in the art will be able to easily confirm the identity in light of the context in which this term is used in this disclosure.

[0058] As used herein, the term "oxo" refers to an oxygen radical linked through a double bond. When the atom bonded to the oxygen is a carbon atom, the bond is a carbon-oxygen double bond, which may be written as -(C=O)- and referred to as a ketone.

[0059] The recitation of a list of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment of a variable or aspect herein includes that embodiment as any single embodiment or in combination with other embodiments or portions thereof.

[0060] Any composition or method provided herein may be combined with any one or more of the other compositions and methods provided herein.

[0061] The following abbreviations are used herein: br Broad signal CDI Carbonyldiimidazole CNS central nervous system CSF cerebrospinal fluid d doublet dd Doublet of Doublets DME Dimethoxyethane DMSO-d6 Dimethyl sulfoxide-d6 DMF Dimethylformamide DNA deoxyribonucleic acid EDTA Ethylenediaminetetraacetic acid EtMgBr Ethyl magnesium bromide EtOAc ethyl acetate GL1 Glucosylceramide (GlcCer) GM3 monosialodihexosylganglioside HPLC High Pressure / Performance Liquid Chromatography HSA Human serum albumin IPA Isopropyl Alcohol J coupling constant LCMS Liquid Chromatography Mass Spectrometry m multiplet ppm (parts per million) rHA Recombinant Human Albumin s singlet TBME Tert-butyl methyl ether THF tetrahydrofuran Tris Tris(hydroxymethyl)aminomethane TWEEN 20 Polysorbate 20 TWEEN80 Polysorbate 80 VOR vestibulo-ocular reflex UPLCMS Ultra-High Performance Liquid Chromatography Mass Spectrometry

[0062] compound The present disclosure relates to quinuclidine compounds for use in therapeutic methods related to the treatment or prevention of the diseases and disorders discussed herein. In all of its various aspects, the present invention relates to a quinuclidine compound (Compound 1) according to formula (I) or a pharmaceutically acceptable salt or prodrug thereof: [ka] (In the formula: R 1 is hydrogen, halogen (e.g., fluorine), cyano, nitro, hydroxy, thio, amino, C 1~6 - alkyl (e.g., methyl or ethyl), C 2~6 -Alkenyl, C 2~6 -alkynyl, C 1~6 -Alkyloxy, C 2~6 -alkenyloxy, and C 2~6 -alkynyloxy, wherein said alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy, or alkynyloxy is optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from halogen, cyano, nitro, hydroxy, thio, or amino; R 2 and R 3is C optionally substituted by one or more (e.g., 1, 2, or 3) halogens; 1~3 -alkyl, or R 2 and R 3 together form a cyclopropyl or cyclobutyl group optionally substituted with one or more (e.g., one or two) halogens; R 4 , R 5 and R 6 is hydrogen, halogen, nitro, hydroxy, thio, amino, C 1~ 6-Alkyl, and C 1~6 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen, hydroxy, cyano, and C 1~6 -optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; A is halogen, hydroxy, thio, amino, nitro, C 1~6 -alkoxy and C 1~6 -a 5- or 6-membered aryl or heteroaryl group (e.g., phenyl or thiazolyl) optionally substituted with 1, 2, or 3 groups independently selected from alkyl.

[0063] In further embodiments of any aspect of the disclosure, the disclosure further relates to compounds such as: 1.1 Compound 1 (wherein R 1 is hydrogen, halogen, cyano, nitro, hydroxy, thio, amino, C 1~6 -Alkyl, C 1~6 -alkyloxy, wherein said alkyl or alkyloxy is optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from halogen, cyano, nitro, hydroxy, thio, or amino; 1.2 Compound 1 (wherein R 1 is hydrogen, halogen, C 1~6 -Alkyl, C 1~6-alkyloxy, wherein said alkyl or alkyloxy is optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from halogen, cyano, nitro, hydroxy, thio, or amino; 1.3 Compound 1 (wherein R 1 is hydrogen, halogen, C 1~4 -alkyl, and C 1~4 -alkyloxy, wherein said alkyl or alkyloxy is optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from halogen, cyano, nitro, hydroxy, thio, and amino; 1.4 Compound 1 (wherein R 1 is hydrogen, halogen, C 1~4 -alkyl, and C 1~4 -alkyloxy, wherein said alkyl or alkyloxy is optionally substituted with one or more (e.g., 1, 2, or 3, or 1 or 2) groups selected from cyano, nitro, hydroxy, thio, and amino; 1.5 Compound 1, wherein R 1 is hydrogen, halogen, and C 1~4 -alkyl, wherein said alkyl is optionally substituted with one or more (e.g., one or two) groups selected from halogen, hydroxy, thio, and amino; 1.6 Compound 1, wherein R 1 is selected from hydrogen, fluorine, methyl, and ethyl, wherein said methyl or ethyl is optionally substituted with one or two groups selected from halogen, hydroxy, thio, and amino; 1.7 Compound 1 (wherein R 1 is selected from hydrogen and methyl, wherein said methyl is optionally substituted with one or two halogens; 1.8 Compound 1 (wherein R 1 is hydrogen); 1.9 Compound 1 or any of 1.1 to 1.8 (wherein R 1 is not attached to the nitrogen atom of the quinuclidine moiety); 1.10 Compound 1 or any of 1.1 to 1.9 (wherein R 2 and R 3 are each independently a C optionally substituted by one or more (e.g., 1, 2, or 3) halogens. 1~3 - alkyl); 1.11 Compound 1.10 (wherein R 2 and R 3 are each independently methyl or ethyl optionally substituted with one or two halogens; 1.12 Compound 1.10 (wherein R 2 and R 3 are each independently selected from methyl and ethyl optionally substituted with one or more fluorines, e.g., 1, 2, 3 or 4 fluorines; 1.13 Compound 1.10 (wherein R 2 and R 3 are each independently methyl substituted with 0, 1, 2, or 3 fluorines; 1.14 Compound 1.10 (wherein R 2 and R 3 are methyl or trifluoro methyl); 1.15 Compound 1.10(R 2 and R 3 are each methyl); 1.16 Compound 1 or any of 1.1 to 1.9 (wherein R 2 and R 3 together form a cyclopropyl or cyclobutyl group optionally substituted with one or more (e.g., one or two) halogens; 1.17 Compound 1.16 (wherein R 2 and R 3 together form a cyclopropyl group); 1.18 Compound 1 or any of 1.1 to 1.9 (wherein R 2 and R 3 are each methyl, or R 2 and R3 together form a cyclopropyl group; 1.19 Compound 1, or any of 1.1-1.9 (wherein R 4 , R 5 and R 6 is hydrogen, halogen, C 1~6 -alkyl, and C 1~6 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen, hydroxy, cyano, and C 1~6 -optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; 1.20 Compound 1 or any of 1.1 to 1.9 (wherein R 4 , R 5 and R 6 is hydrogen, halogen, C 1~3 -alkyl, and C 1~3 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen, hydroxy, cyano, and C 1~3 -optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; 1.21 Compound 1.19 (wherein R 4 , R 5 and R 6 is hydrogen, halogen, C 1~3 -alkyl, and C 1~3 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen, cyano, and C 1~3 - optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; 1.22 Compound 1.19 (wherein R 4 , R 5 and R 6 is hydrogen, halogen, C 1~3 -alkyl, and C 1~3 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen and C 1~3- optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; 1.23 Compound 1.19 (wherein R 4 , R 5 and R 6 is a halogen, C 1~3 -alkyl, and C 1~3 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen and C 1~3 - optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; 1.24 Compound 1 or any of 1.19 to 1.23 (wherein R 4 is hydrogen, halogen, C 1~3 -alkyl, and C 1~3 -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen and C 1~3 - optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; 1.25 Compound 1.24(R 4 is a halogen (e.g., fluorine), C 1~3 -alkyl (e.g., methyl), and C 1~3 -alkyloxy (e.g., methoxy or ethoxy), wherein said alkyl or alkyloxy is selected from halogen and C 1~3 - optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy (e.g., methoxy or ethoxy); 1.26 Compound 1.25(R 4 is a halogen (e.g., fluorine) and C 1~3 -alkyloxy (e.g., methoxy or ethoxy), wherein said alkyloxy is selected from halogen and C 1~3 - optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy (e.g., methoxy or ethoxy); 1.27 Compound 1.26 (wherein R 4 is fluorine or halogen and C 1~3 -a C optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy (e.g., methoxy), 1~3 - alkyloxy (e.g., ethoxy); 1.28 Compound 1.26 (wherein R 4 is fluorine or one or more (e.g., 1, 2, or 3) C 1~3 -ethoxy optionally substituted by alkyloxy (e.g., methoxy); 1.29 Compound 1 or any of 1.19 to 1.28 (wherein R 6 is hydrogen); 1.30 Compound 1 or any of 1.19 to 1.28 (wherein R 5 and R 6 are hydrogens); 1.31 Compound 1 or any of 1.19 to 1.28 (R 5 and R 6 are hydrogen, and R 4 is fluorine or halogen and C 1~3 -C optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy (e.g., methoxy), 1~3 - alkyloxy (e.g., ethoxy); 1.32 Compound 1.31 (wherein R 5 and R 6 are hydrogen, and R 4 is fluorine or one or more (e.g., 1, 2, or 3) C 1~3 -ethoxy optionally substituted by alkyloxy (e.g., methoxy); 1.33 Compound 1.32 (wherein R 5 and R 6 are hydrogen, and R 4is fluorine, or ethoxy substituted with methoxy (e.g., 2-methoxyethoxy); 1.34 Compound 1.32 (wherein R 4 is fluorine or 2-methoxyethoxy); 1.35 Compound 1, or any of 1.1 to 1.34, where R 4 , R 5 and R 6 at least one of which is not hydrogen); 1.36 Compound 1 or any of 1.1 to 1.35 (wherein R 6 is hydrogen and R 4 and R 5 are located at the 2-, 4-, or 6-position of the phenyl ring to which they are attached (i.e., ortho or para to the A substituent); 1.37 Compound 1 or any of 1.1 to 1.35 (wherein R 6 is hydrogen and R 4 and R 5 are independently located (relative to the A substituents) at the 2- and 3-positions (i.e., adjacent ortho and meta), at the 3- and 4-positions (i.e., adjacent meta and para), or at the 3- and 5-positions (i.e., meta) of the phenyl ring to which they are attached; 1.38 Compound 1, or any of 1.1-1.35 (wherein R 6 is hydrogen and R 4 and R 5 are located at the 3 and 5 positions (i.e., meta) of the phenyl ring to which they are attached (relative to the A substituent); 1.39 Compound 1 or any of 1.1 to 1.35 (wherein R 5 and R 6 is hydrogen and R 4 is located at the 2-, 3-, or 4-position of the phenyl ring to which it is attached (e.g., ortho, meta, or para to the A substituent); 1.40 Compound 1 or any of 1.1 to 1.35 (wherein R 5 and R 6 is hydrogen and R 4is located at the 2- or 4-position of the phenyl ring to which it is attached (e.g., ortho or para to the A substituent); 1.41 Compound 1 or any of 1.1 to 1.35 (wherein R 5 and R 6 is hydrogen and R 4 is located at the 4-position of the phenyl ring to which it is attached (e.g., para to the A substituent); 1.42 Compound 1 or any of 1.1 to 1.35 (wherein R 4 , R 5 and R 6 are not hydrogen, and each R 4 , R 5 , and R 6 are independently located at the 2-, 4-, or 6-position of the phenyl ring to which they are attached (i.e., ortho or para to the A substituent); 1.43 Compound 1 or any of 1.1 to 1.42 (wherein R 4 is located at the 4-position of the phenyl ring to which it is attached (i.e., para to the A substituent); 1.44 Compound 1 or any of 1.1-1.43, wherein A is a 6-membered aryl group , a 5-membered heteroaryl group (e.g., containing 1, 2, or 3 heteroatoms in the heteroaryl ring independently selected from N, O, and S), or a 6-membered heteroaryl group (e.g., containing 1, 2, or 3 nitrogen atoms in the heteroaryl ring); 1.45 Compound 1.44, where A is a 6-membered aryl group or a 5-membered heteroaryl group (e.g., containing 1, 2, or 3 heteroatoms in the heteroaryl ring independently selected from N, O, and S), optionally wherein the 5-membered heteroaryl group contains 1 or 2 heteroatoms selected from N and S (e.g., 1 N and / or 1 S); 1.46 Compound 1.44 or 1.45, wherein A is selected from the group consisting of phenyl, furyl, thienyl, thiazolyl, pyrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl, imidazolyl, oxadiazolyl, and thiadiazolyl; 1.47 Compound 1.46, wherein A is selected from the group consisting of phenyl, thienyl, thiazolyl, pyrrolyl, and imidazolyl; 1.48 Compound 1.46, wherein A is selected from the group consisting of phenyl and thiazolyl, e.g., 2-thiazol-4-yl or 4-thiazol-2-yl; 1.49 Compound 1 or any of 1.1 to 1.48 (wherein A is unsubstituted) 1.50 Compound 1 or any of 1.1 to 1.48 (wherein A is halogen, hydroxy, thio, amino, nitro, C 1~6 Alkoxy, and C 1~6 substituted with one or more (e.g., 1, 2, or 3) groups independently selected from alkyl (e.g., methyl); 1.51 Compound 1.50, where A is one halogen (e.g., fluorine) or C 1~6 thiazolyl substituted with alkyl (e.g., methyl); 1.52 Compound 1.50 (wherein A is a halogen (e.g., fluorine) and C 1~6 phenyl substituted with one, two, or three groups independently selected from alkyl (e.g., methyl); 1.53 Compound 1.52 (wherein A is phenyl substituted with one or two fluorine or methyl groups); 1.54 Compound 1 or any of 1.1-1.53 ​​(wherein the two groups attached to the A substituent (i.e., the phenyl ring (-(CHR 4 R 5 R 6 )) and -C(R 2 R 3 )-groups) are positioned in a 1,2, 1,3, or 1,4 relationship to each other (i.e., ortho, meta, or para); 1.55 Compound 1.54 (wherein the two groups attached to the A substituents are positioned in a 1,3 relationship (i.e., meta) to each other); 1.56 Compound 1.54 (wherein the two groups attached to the A substituent are positioned in a 1,4 relationship (i.e., para) to each other); 1.57 Any of compounds 1.54 through 1.56, wherein the A substituent is a 5-membered heteroaryl group and the two groups attached to the A substituent (i.e., the phenyl ring (-(CHR 4 R 5 R 6 )) or the-C(R 2 R 3 )-groups) are bonded to a carbon atom of the heteroaryl ring, and optionally both such groups are bonded to a carbon atom of the heteroaryl ring;

[0064] 1.58 Compound 1, or any of 1.1-1.57, wherein the compound of formula (I) may be represented by one or more of the following structures: [ka] [ka] [ka]

[0065] 1.59 Compound 1, or any of 1.1 to 1.58 (wherein the compound of formula (I) or any of formulas (II) to (XII) has the (S) configuration); 1.60 Compound 1, or any of 1.1 to 1.58 (wherein the compound of formula (I) or any of formulas (II) to (XII) has the (R) configuration); 1.61 Compound 1, or any of 1.1-1.60, wherein the compound of formula (I) or any of formulas (II) to (XII) has an enantiomeric excess (e.g., of the (S) configuration) of at least 90%, e.g., at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9%; 1.62 Compound 1, or any of 1.1-1.58 (wherein the compound of formula (I) or any of formulas (II) to (XII) is a racemate (i.e., approximately a 50:50 ratio of enantiomers) or a mixture of enantiomers in some other ratio (e.g., less than or greater than 50:50);

[0066] 1.63 Compound 1, or any of 1.1 to 1.62, wherein the compound of formula (I) is selected from the group consisting of the following tables: [Table 1-1] [Table 1-2]

[0067] 1.64 Compound 1, or any of 1.1-1.63, wherein the compound is selected from quinuclidin-3-yl(2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate, (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, and (S)-quinuclidin-3-yl(2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate; 1.65 Compound 1, or any of 1.1 to 1.63 (wherein the compound is quinuclidin-3-yl(2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate); 1.66 Compound 1 or any of 1.1 to 1.63 (wherein the compound is quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, for example, (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate); 1.67 Compound 1 or any of 1.1 to 1.66 (wherein the compound of formula (I), or any of formulas (II) to (XII), is in the form of a free base); 1.68 Compound 1, or any of 1.1 to 1.66 (wherein the compound of formula (I), or any of (II) to (XII), is in the form of a pharmaceutically acceptable salt); 1.69 Compound 1.68 (wherein said salt form is an acid addition salt form); 1.70 Compound 1.69, wherein the acid addition salt form is a salt selected from hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, hydroxysuccinate, malate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, and pamoate; 1.71 Compound 1.70, wherein the acid addition salt form is selected from hydrochloride, hydroxysuccinate (e.g., 2-hydroxysuccinate), and malate; 1.72 Compound 1.68 (wherein said salt form is a base addition salt form); 1.73 Compound 1, or any of 1.1-1.72, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazolinone)) in the form of a malate. (4-yl)propan-2-yl)carbamate); 1.74 Compound 1, or any of 1.1-1.73, wherein the compound of formula (I), or any of (II) to (XII), is in the form of a prodrug as described herein; 1.75 Compound 1, or any of 1.1 to 1.74, wherein the compound of formula (I), or any of (II) to (XII), is in the form of a hydrate, solvate and / or polymorph.

[0068] salt The compounds disclosed herein, such as compounds 1 or any of 1.1 through 1.75, are essentially basic and can generally form a variety of different salts with various inorganic and / or organic acids. While such salts are generally pharmaceutically acceptable for administration to animals and humans, it is often practically desirable to first isolate the compound from the reaction mixture as a pharmaceutically unacceptable salt, then simply convert the latter back to the free base compound by treatment with an alkaline reagent, and then convert the free base to a pharmaceutically acceptable acid addition salt. Acid addition salts of basic compounds may be readily prepared using conventional techniques, for example, by treating the basic compound with a substantially equivalent amount of a selected inorganic or organic acid in an aqueous solvent medium or in a suitable organic solvent, such as methanol or ethanol. Careful evaporation of the solvent yields the desired solid salt. The compounds disclosed herein are positively charged, including, for example, quaternary ammonium, and can also form salts with the anionic components of various inorganic and / or organic acids.

[0069] Acids which may be used to prepare pharmaceutically acceptable salts of quinuclidine compounds are those capable of forming non-toxic acid addition salts, e.g., salts containing pharmacologically acceptable anions, such as chloride, bromide, iodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate or acid citrate, tartrate or bitartrate, succinate, malate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, and pamoate [i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)] salts.

[0070] Compounds disclosed herein that are acidic in nature, e.g., compounds containing a thiol moiety, can generally form a variety of different salts with various inorganic and / or organic bases. While such salts are generally pharmaceutically acceptable for administration to animals and humans, it is often practically desirable to first isolate the compound from the reaction mixture as a pharmaceutically unacceptable salt, then simply convert the latter back to the free acid compound by treatment with an acidic reagent, and then convert the free acid to a pharmaceutically acceptable base addition salt. These base addition salts may be readily prepared using conventional techniques, for example, by treating the corresponding acidic compound with an aqueous solution containing the desired pharmacologically acceptable cation, and then evaporating the resulting solution to dryness, e.g., under reduced pressure. Alternatively, they may be prepared by mixing lower alkanol solutions of the acidic compound with the desired alkali metal alkoxide, and then evaporating the resulting solution to dryness in the same manner as described above. In either case, stoichiometric amounts of reagents may be used to ensure completeness of the reaction and maximize the yield of the desired solid salt product.

[0071] Bases that may be used to prepare pharmaceutically acceptable base addition salts of quinuclidine compounds are those capable of forming non-toxic base addition salts, for example, salts containing pharmacologically acceptable cations, such as alkali metal cations (e.g., potassium and sodium), alkaline earth metal cations (e.g., calcium and magnesium), ammonium or other water-soluble amines, e.g., N-methylglucamine (meglumine), lower alkanolammonium, and other organic amine bases.

[0072] In one embodiment, the pharmaceutically acceptable salt is a succinate salt. In another embodiment, the pharmaceutically acceptable salt is a 2-hydroxysuccinate salt, for example, (S)-2-hydroxysuccinate salt. In another embodiment, the pharmaceutically acceptable salt is a hydrochloride salt (i.e., a salt with HCl). In another embodiment, the pharmaceutically acceptable salt is a malate salt.

[0073] Prodrug The present disclosure further encompasses prodrugs of compounds 1 and 1.1 to 1.75. The pharmaceutically acceptable prodrugs disclosed herein are derivatives of quinuclidine compounds that can be converted in vivo to the quinuclidine compounds described herein. The prodrugs may themselves have some activity, for example, become pharmaceutically active in vivo when subjected to solvolysis under physiological conditions or enzymatic degradation. Methods for preparing prodrugs of the compounds described herein will be apparent to those skilled in the art based on the present disclosure.

[0074] In one embodiment, the carbamate moiety of the quinuclidine compound is modified. For example, the carbamate moiety of the quinuclidine compound may be modified by adding water and / or one or two aliphatic alcohols. In this case, the carbon-oxygen double bond of the carbamate moiety adopts what can be considered a hemiacetal or acetal functionality. In one embodiment, the carbamate moiety of the quinuclidine compound may be modified by adding an aliphatic diol, such as 1,2-ethanediol.

[0075] In one embodiment, one or more of the hydroxy, thio, or amino groups in the quinuclidine compound are modified. For example, one or more of the hydroxy, thio, and / or amino groups in the quinuclidine compound may be modified to form an acid derivative, such as an ester, a thioester (or a thiol ester), and / or an amide. The acid derivative may be formed, for example, by reacting a quinuclidine compound containing one or more hydroxy, thio, or amino groups with an acetylating agent. Examples of acetylating agents include anhydrides, such as acetic anhydride, acid chlorides, such as benzyl chloride, and bicarbonates, such as di-tert-butyl dicarbonate.

[0076] stereochemistry The present disclosure further encompasses stereoisomers and mixtures of stereoisomers of compounds 1 and 1.1 through 1.75. Stereoisomers (e.g., cis and trans isomers) and all optical isomers (e.g., R- and S-enantiomers) of the compounds disclosed herein, as well as racemic, diastereomeric, and other mixtures of such isomers, are within the scope of the present disclosure.

[0077] In one embodiment, the quinuclidine-3-yl group of the quinuclidine compounds defined herein has the R-configuration. Thus, the quinuclidine compound may be selected from the group consisting of compounds of formula (Ia) to (XIIa), and pharmaceutically acceptable salts and prodrugs thereof: [ka] [ka] [ka]

[0078] In another embodiment, the quinuclidine-3-yl group of the quinuclidine compounds defined herein has the S-configuration. Thus, the quinuclidine compound may be selected from the group consisting of compounds of formula (Ib) to (XIIb), and pharmaceutically acceptable salts and prodrugs thereof: [ka] [ka] [ka]

[0079] In one embodiment, the quinuclidine compound is a compound of Formula (Xb) or a pharmaceutically acceptable salt or prodrug thereof. In another embodiment, the quinuclidine compound is a compound of Formula (XIIb) or a pharmaceutically acceptable salt or prodrug thereof.

[0080] In one embodiment, the quinuclidine-3-yl group of the quinuclidine compound defined herein exists as a mixture of isomers having R- and S-configuration.For example, the quinuclidine compound may be a mixture of compounds selected from the group consisting of compounds of formula (Ia) and (Ib), (IIa) and (IIb), (IIIa) and (IIIb), (IVa) and (IVb), (Va) and (Vb), (VIa) and (VIb), (VIIa) and (VIIb), (VIIIa) and (VIIIb), (IXa) and (IXb), (Xa) and (Xb), (XIa) and (XIb), and (XIIa) and (XIIb), and pharmaceutically acceptable salts and prodrugs thereof.In one embodiment, the quinuclidine compound exists as a racemic mixture, for example, the R- and S-isomers of the quinuclidine-3-yl group are present in approximately equal amounts. In another embodiment, the quinuclidine compound exists as a mixture of isomers having R- and S-configuration, and the R- and S-isomers are present in different amounts. In one embodiment, the S-isomer is present in an enantiomeric excess of at least about 5%, 10%, 25%, 40%, 70%, 80%, 90%, 95%, 97%, 98%, or 99%, for example, about 100%. In another embodiment, the R-isomer is present in an enantiomeric excess of at least about 5%, 10%, 25%, 40%, 70%, 80%, 90%, 95%, 97%, 98%, or 99%, for example, about 100%.

[0081] Methods for preparing enantiomerically enriched and / or enantiopure quinuclidine compounds will be apparent to those skilled in the art based on the present disclosure.

[0082] The compounds disclosed herein may exist in several tautomeric forms, including enol and imine forms, and keto and enamine forms, as well as geometric isomers, and mixtures thereof. Tautomers exist as a mixture of tautomeric sets in solution. In solid form, one tautomer usually predominates. Although one tautomer may be described, all tautomers are within the scope of this disclosure.

[0083] Atropisomers are also within the scope of this disclosure. Atropisomers refer to compounds that can be decomposed into isomers with restricted rotation.

[0084] Other forms The present disclosure further encompasses hydrates, solvates, and polymorphs of compounds 1 and 1.1 to 1.75. Pharmaceutically acceptable hydrates, solvates, and polymorphs of the quinuclidine compounds described herein are within the scope of the present disclosure. The compound may be in amorphous form and / or in one or more crystalline forms.

[0085] Isotopically labeled compounds are also within the scope of the present disclosure.As used herein, " isotope-labeled compounds " refers to the compounds disclosed herein, including pharmaceutical salts and their prodrugs as described herein, wherein one or more atoms are replaced by atoms with atomic mass or mass number that is different from the atomic mass or mass number that is found in nature.The examples of isotopes that can be incorporated into compounds disclosed herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, for example, respectively: 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, and 36 There is Cl.

[0086] Medical indication The quinuclidine compounds described herein and the pharmaceutical compositions comprising them are useful, for example, in therapy, particularly in the therapeutic treatment of neurological disorders, including dementia and gait disorders, in patients with diseases such as Gaucher's disease.The subjects to be treated according to the methods described herein include vertebrates, for example mammals.In certain embodiments, the mammal is a human patient.

[0087] As mentioned above, one of the characteristics of glycogen storage diseases is the abnormal accumulation of various glycolipids or glycosphingolipids in the body's cells. This accumulation is responsible for the observable symptoms and signs of the disease and also serves as a diagnostic marker that demonstrates the presence and / or progression of the disease. As used herein, the phrase "significant accumulation" with respect to the measurement of GL-3, GL-1, and other biomarkers in plasma, skin, or other soft tissues means accumulation of more than 25% above the maximum normal concentration of the compound. In some embodiments, "significant accumulation" means accumulation of more than 50% above the maximum normal concentration of the compound.

[0088] In a first aspect, the present invention provides a method (Method 1) for treating or preventing cognitive impairment and / or gait abnormalities, including ataxia, associated with a lysosomal storage disease in a subject, such as a subject in need thereof, the method comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or Compounds 1 or 1.1-1.75. Also provided are quinuclidine compounds described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or Compounds 1 or 1.1-1.75, for use in a method for treating or preventing cognitive impairment and / or gait abnormalities, including ataxia, associated with a lysosomal storage disease in a subject in need thereof, e.g., for use in Method 1 or 1.1-1.64. Further provided is the use of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compound 1 or any of 1.1 to 1.75, in the manufacture of a medicament for use in a method of treating or preventing cognitive impairment and / or gait abnormalities, including ataxia, associated with a lysosomal storage disease in a subject in need thereof, e.g., in the manufacture of a medicament for use in any of Methods 1 or 1.1-1.64.

[0089] In certain further embodiments of Method 1, the present disclosure provides: 1.1. Method 1, comprising administering to a subject an effective amount of a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compound 1 or any of 1.1 to 1.75; 1.2. An effective amount of Compound 1 or any one of Compounds 1.1 to 1.75 or Method 1, comprising administering to a subject: 1.3. Any of Methods 1 or 1.1-1.2, comprising administering to a subject an effective amount of a pharmaceutical composition comprising a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compound 1 or any of 1.1 to 1.75; 1.4. Any of Methods 1 or 1.1-1.2, comprising administering to a subject an effective amount of a pharmaceutical composition comprising Compound 1 or any one or more of Compounds 1.1 to 1.75; 1.5. Method 1.3 or 1.4, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient described herein; 1.6. Any of Methods 1 or 1.1-1.5, comprising administering a pharmaceutical dosage form comprising an effective amount of a compound or an effective amount of a pharmaceutical composition; 1.7. Method 1.6, wherein the dosage form is an oral dosage form (e.g., a pill, capsule, caplet, tablet, dragee, powder, granules, film, lozenge, or liquid); 1.8. Method 1.7, wherein the dosage form is a chewable tablet; 1.9. The dosage form is a parenteral dosage form (e.g., when the pharmaceutical composition is formulated for injection), method 1.6; 1.10. The injection is intravenous, intramuscular, intrathecal, or subcutaneous, optionally sterile, Method 1.9; 1.11. Method 1.6, wherein the dosage form is a topical or rectal dosage form; 1.12. Method 1.6, wherein the dosage form is an intranasal dosage form (e.g., an aerosol); 1.13. Any of Methods 1 or 1.1 through 1.12, further comprising concurrently administering in a patient in need thereof a second active agent described herein, e.g., a second compound capable of treating or preventing cognitive impairment and / or gait abnormalities; 1.14. Method 1.13, wherein the second active agent is administered in the same pharmaceutical composition or dosage form as the quinuclidine compound; 1.15. Methods 1.13 or 1.14, wherein the second active agent is a GCS inhibitor (e.g., miglustat or eliglustat); 1.16. Method 1 or any of 1.1 to 1.15, wherein the subject is a mammal; 1.17. Method 1.16, wherein the subject is a primate; 1.18. Method 1.17, wherein the subject is a human; 1.19. The ataxia is cerebellar ataxia, method 1 or any of 1.1 to 1.18; 1.20. Ataxia presents with symptoms selected from gait instability, asthenia, insynergia, slowed reaction time, dyschronometria, dysarthria, dysphagia, hypotonia, dysmetria, hypometria, hypermetria, dyspraxia of antagonistic movements, speech slurring, vocal tremor, ataxic breathing, postural instability, and combinations thereof, for example, the predominant ataxic deficit is gait instability, Method 1.19; 1.21. Subjects will complete the Scale for Assessment and Rating of Ataxia at the start of treatment with the method. having a baseline ataxia of at least 0.5 on the Standardized Ataxia Assessment (SARA) scale, e.g., a baseline SARA score of at least 1, or at least 2, or at least 3, or at least 4, or at least 5, or at least 10, or at least 20, Method 1.19 or 1.20; 1.22. The cognitive impairment is dementia, method 1 or any of 1.1 to 1.21; 1.23. Dementia may be present, for example, with a TMT-A score of more than 30 seconds, or more than 45 seconds, or more than 60 seconds and / or a score of more than 70 seconds, or more than 90 seconds, or more than 120 seconds, or more than 150 seconds, or show signs of deficits in visual search speed, processing scanning speed, mental flexibility, and / or executive function as evidenced by TMT-B of greater than 180 seconds or greater than 40 seconds, or greater than 60 seconds, or greater than 90 seconds, or greater than 120 seconds, Method 1.22; 1.24. The subject has Gaucher disease type 3, method 1, or any of 1.1-1.23; 1.25. The subject has Niemann-Pick disease type C, and any of Methods 1 or 1.1-1.24; 1.26. The subject has GM2-gangliosidosis (e.g., Tay-Sachs disease, Sandhoff disease, or GM2 gangliosidosis AB variant), according to Method 1 or any of Methods 1.1-1.24; 1.27. The subject has been diagnosed with a mutation in the gene GBA1, according to method 1 or any of methods 1.1 to 1.24; 1.28. Method 1 or any of 1.1-1.24, wherein the subject has been diagnosed with a mutation in the genes NPC1 and / or NPC2; 1.29. Method 1, or any of 1.1-1.24, wherein the subject has been diagnosed with a mutation in the gene HEXA (encoding hexosaminidase A) and / or a mutation in the gene HEXB (encoding hexosaminidase B) and / or a mutation in the gene GM2A (encoding GM2 ganglioside activator protein); 1.30. The subject has been diagnosed with Parkinson's disease, either Method 1 or 1.1-1.29; 1.31. Method 1, or any of 1.1-1.30, wherein the subject is undergoing concurrent treatment with an enzyme replacement therapy (ERT), e.g., with glucocerebrosidase (e.g., imiglucerase, velaglucerase, or taliglucerase), optionally each such enzyme being a recombinant enzyme; 1.32. Method 1.31, wherein the subject is undergoing concurrent treatment with one or more of imiglucerase, velaglucerase (e.g., velaglucerase alfa), and taliglucerase (e.g., taliglucerase alfa); 1.33. The subject is receiving concurrent treatment with imiglucerase, Method 1.32; 1.34. The subject is receiving concurrent treatment with imiglucerase (1 unit of imiglucerase is the amount of enzyme that catalyzes the hydrolysis of 1 micromole of the synthetic substrate p-nitrophenyl-β-D-glucopyranoside per minute at 37°C) at a dosage of 2.5 units / kg body weight to 80 units / kg body weight every 1 to 3 weeks, e.g., 40 to 60 units / kg body weight every 2 weeks, Method 1.33; 1.35. Method 1.34, wherein the subject's dosage of imiglucerase in each administration (e.g., every 1 to 3 weeks, e.g., every 2 weeks) is administered as an intravenous (IV) infusion over 1 to 3 hours (e.g., 1 to 2 hours); 1.36. Any of Methods 1 or 1.1-1.35, wherein the subject has been administered an enzyme replacement therapy (e.g., imiglucerase, velaglucerase, and / or taliglucerase) prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 1.37. Method 1.36, wherein the subject has been administered an imiglucerase therapeutic agent for at least 6 months, e.g., at least 12 months (1 year), or at least 18 months, or at least 2 years, or at least 3 years, prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or compound 1 or any of 1.1-1.75). 1.38. The subject is a compound of formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or a compound 1 or 1.1 to 1.75 have been receiving imiglucerase treatment at a stable dose for at least 6 months prior to initiating treatment with the compound according to either method 1.36 or 1.37; 1.39. Any of Methods 1 or 1.1-1.38, further comprising the step of transitioning the subject from an ERT therapeutic agent (e.g., imiglucerase, velaglucerase, or taliglucerase) to treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 1.40. Any of Methods 1 or 1.1-1.39, wherein the subject's hemoglobin level is at least 11 g / dL for women and at least 12 g / dL for men; 1.41. The subject's platelet count is at least 100,000 per cubic millimeter, as described in Method 1 or any of Methods 1.1 through 1.40; 1.42. Subject has a spleen volume less than 10 times normal (10 MN) and / or a liver volume less than 1.5 MN, either Method 1 or 1.1-1.41; 1.43. The subject has been diagnosed with a co-occurring dementia, e.g., Alzheimer's disease or Parkinson's disease, according to any of Methods 1 or 1.1-1.42; 1.44. Method 1, or any of 1.1-1.43, wherein the subject is at least 18 years of age (e.g., 18-30 years of age) at the start of treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 1.45. The subject's glucosylceramide (GL1) concentration is 4.4 to 11.1 ng / mL in cerebrospinal fluid (CSF) and 4.9 to 8.3 μg / mL in plasma, according to any of Methods 1 or 1.1 to 1.44; 1.46. The subject's glucosylsphingosine (lyso-GL1) concentration is 20.1-67.6 pg / mL in CSF and 8.8-159.0 ng / mL in plasma, either Method 1 or 1.1-1.45; 1.47. Method 1 or any of 1.1-1.46, wherein the subject is administered a daily dose of a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compound 1 or any of 1.1-1.75) selected from about 1 mg to about 150 mg, e.g., 5 to 50 mg, or 10 to 40 mg, or 10 to 30 mg, or 10 to 20 mg, or 20 to 30 mg, or 30 to 40 mg, or 40 to 50 mg, or 5 to 25 mg, or 20 to 50 mg, or 5 to 15 mg, or 15 to 30 mg, or about 15 mg, or 2, 5, 15, 25, 50, 100, or 150 mg; 1.48. Method 1, or any of 1.1-1.47, wherein the subject is a human adult patient, e.g., between 18 and 80 years of age, e.g., between 18 and 60 years of age, or between 18 and 40 years of age, or between 18 and 30 years of age, or between 18 and 25 years of age; 1.49. Method 1, or any of 1.1-1.47, wherein the subject is a human pediatric patient, e.g., 0 to 18 years of age, e.g., 1 to 15 years of age, or 1 to 5 years of age, or 5 to 10 years of age, or 10 to 15 years of age, or 10 to 18 years of age; 1.50. Any of Methods 1 or 1.1-1.49, which is effective to reduce the SARA ataxia scale by at least 0.5, e.g., to reduce the SARA score by at least 1, or at least 2, or at least 3, or at least 5, or at least 10; or the method is effective to reduce the SARA score between 0.00 and 3.00, or between 0.00 and 2.00, or between 0.00 and 1.50, or between 0.00 and 1.00, or between 0.00 and 0.50. 1.51. For example, a reduction in the time taken to complete the Trail Making Test (TMT), TMT-A and / or TMT-B, a reduction in the difference between TMT-A and TMT-B times (TMT-A-TMT-B), for example, by at least 10%, or or is effective in improving cognitive performance or reducing cognitive deficits as measured by a decrease of at least 20%, or at least 30%, or at least 40%, or at least 50% (e.g., a 5-20% decrease in TMT-A and / or a 25-30% decrease in TMT-B and / or a 25-30% decrease in [TMT-A-TMT-B]), either Method 1 or 1.1-1.50; 1.52. Any of Methods 1 or 1.1-1.51, wherein the method results in at least a 30%, e.g., at least a 40%, at least a 50%, at least a 60%, or at least a 70% decrease in glucosylceramide levels in the CSF and / or in the plasma after 6 months of treatment; 1.53. Any of Methods 1 or 1.1-1.52, wherein the method results in an increase in glucosylsphingosine concentration in CSF and / or plasma of at least 30%, e.g., at least 40%, at least 50%, at least 60%, or at least 70% after 6 months of treatment; 1.54. Any of Methods 1 or 1.1-1.53, wherein the method results in a statistically or clinically unchanged modified Severity Scoring Tool (mSST) value for neurological disease after 6 months of treatment; 1.55. Any of Methods 1 or 1.1-1.54, wherein the method results in increased blood flow in the brain (e.g., in one or more of the frontal lobe, occipital lobe, parietal lobe, or temporal lobe), e.g., as shown by fMRI imaging; 1.56. Any of Methods 1 or 1.1-1.54, wherein the method results in increased nodal connectivity in the brain (e.g., between the posterior and anterior sides of the brain, and / or between occipito-parietal structures and frontal, temporal, and / or limbic structures), e.g., as shown by fMRI imaging; 1.57. Method 1, or any of 1.1 to 1.56, wherein a compound according to Formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or Compound 1 or any of 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a parenteral or non-parenteral route; 1.58. The route of administration is oral (enteral), Method 1.57; 1.59. Method 1.57, in which the route of administration is parenteral, e.g., by injection, e.g., by intravenous injection; 1.60. Method 1, or any of 1.1 to 1.59, wherein a compound according to Formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or Compound 1 or any of 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by local administration, e.g., topical administration; 1.61. Method 1, or any of 1.1 through 1.60, wherein the compound is (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate or quinuclidin-3-yl (2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate; 1.62. The compound dosage is 15 mg / day orally, Method 1.61; 1.63. The compound dosage is 15 mg / day in a single oral dose, Method 1.62; 1.64. Method 1, or any of 1.1-1.63, wherein the subject is administered a single daily dose of 5 mg, 10 mg, 15 mg, or 20 mg of a compound, e.g., (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate.

[0090] In some embodiments of the present disclosure, a subject or patient has been diagnosed with a particular disease or disorder and has also been diagnosed with a particular genetic mutation, e.g., one known to result in the disease or disorder in question, but the disease or disorder in a particular patient is not known to result in the disease or disorder in question. However, it is often not possible to prove that a specific mutation that a person has been diagnosed with is caused by the mutation. When used in this way, the term "diagnosed to have a specific genetic mutation" means that a subject or patient has been tested, for example, by DNA or RNA sequencing, protein profiling, or other suitable means, and has been found to have the mutation in question. However, as will be further explained below, many genetic diseases and disorders may have multiple genetic causes (e.g., mutations), and a patient may have multiple mutations that may each be sufficient to cause a disease or disorder under some circumstances, and it is not possible to prove that a specific mutation causes a specific disease or disorder in a specific patient.

[0091] Methods according to Method 1 and subsequent methods may be beneficial for subjects who have been diagnosed with a lysosomal storage disease, such as Gaucher Type 3 or Niemann-Pick Type C, but who have not yet experienced the cognitive and / or ataxic symptoms associated with the condition. Methods according to Method 1 and subsequent methods may also be beneficial for subjects who are at risk of developing a lysosomal storage disease, such as Gaucher Type 3 or Niemann-Pick Type C, for example, due to a mutation in the subject or the subject's family known to cause such a disease. Thus, in some embodiments of the methods described herein, the subject has been diagnosed as being at risk for developing the disease or disorder, and the method prevents or delays the onset and / or development of cognitive and / or ataxic symptoms of the disease or disorder in the subject. In some embodiments, the subject has been diagnosed as being at risk for developing the disease or disorder due to having a mutation in a gene described herein.

[0092] In a second aspect, the present invention provides a method (Method 2) for enhancing neuronal connectivity in the brain in a subject, such as a subject in need thereof, the method comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compounds 1 or any of 1.1-1.75. Also provided are quinuclidine compounds described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compounds 1 or any of 1.1-1.75, for use in Method 2 or 2.1-2.67 for enhancing neuronal connectivity in the brain of a subject. Further provided is the use of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compound 1 or any of 1.1-1.75, in the manufacture of a medicament for enhancing neuronal connectivity in the brain of a subject, e.g., in the manufacture of a medicament for use in Method 2 or 2.1-2.67.

[0093] In certain further embodiments of Method 2, the present disclosure provides: 2.1. Method 2, comprising administering to a subject an effective amount of a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compound 1 or any of 1.1 to 1.75; 2.2. Method 2, comprising administering to a subject an effective amount of Compound 1 or any one or more of Compounds 1.1 to 1.75; 2.3. Any of Methods 2 or 2.1-2.2, comprising administering to a subject an effective amount of a pharmaceutical composition comprising a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compound 1 or any of 1.1 to 1.75; 2.4. Method 2 or any of Methods 2.1-2.2, comprising administering to a subject an effective amount of a pharmaceutical composition comprising Compound 1 or any one or more of Compounds 1.1 to 1.75. Either; 2.5. Method 2.3 or 2.4, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient described herein; 2.6. Any of Methods 2 or 2.1-2.5, comprising administering a pharmaceutical dosage form comprising an effective amount of a compound or an effective amount of a pharmaceutical composition; 2.7. Method 2.6, wherein the dosage form is an oral dosage form (e.g., a pill, capsule, caplet, tablet, dragee, powder, granules, film, lozenge, or liquid); 2.8. Method 2.7, wherein the dosage form is a chewable tablet; 2.9. The dosage form is a parenteral dosage form (e.g., when the pharmaceutical composition is formulated for injection), Method 2.6; 2.10. Method 2.9, in which the injection is intravenous, intramuscular, intrathecal, or subcutaneous, and optionally is a sterile injection; 2.11. Method 2.6, wherein the dosage form is a topical or rectal dosage form; 2.12. Method 2.6, wherein the dosage form is an intranasal dosage form (e.g., an aerosol); 2.13. Any of Methods 2 or 2.1-2.12, further comprising simultaneously administering a second active agent, e.g., a second compound described herein that can reduce glycosylceramide levels in a patient in need thereof; 2.14. Method 2.13, wherein the second active agent is administered in the same pharmaceutical composition or dosage form as the quinuclidine compound; 2.15. Method 2.13 or 2.14, wherein the second active agent is a GCS inhibitor (e.g., miglustat or eliglustat); 2.16. Method 2 or any of 2.1-2.15, wherein the subject is a mammal; 2.17. Method 2.16, wherein the subject is a primate; 2.18. Method 2.17, wherein the subject is a human; 2.19. Method 2, or any of 2.1-2.18, wherein the subject has a symptom selected from ataxia, e.g., gait instability, asthenia, insynergia, slowed reaction time, dyschronometry, dysarthria, dysphagia, hypotonia, dysmetria, hypometria, hypermetria, dyspraxia of antagonistic movements, speech disorder, vocal tremor, ataxic breathing, postural instability, and combinations thereof, e.g., wherein the predominant ataxic deficit is gait instability; 2.20. Subjects will complete the Scale for Assessment and Rating of Ataxia at the start of treatment with the method. Method 2.19, having a baseline ataxia of at least 0.5 on the State of Ataxia (SARA) scale, e.g., a baseline SARA score of at least 1, or at least 2, or at least 3, or at least 4, or at least 5, or at least 10, or at least 20; 2.21. The subject has cognitive impairment (e.g., dementia), Method 2, or any of Methods 2.1-2.20; 2.22. Cognitive impairment is dementia, Method 2.21; 2.23. Dementia is characterized by signs of deficits in visual search speed, processing scanning speed, mental flexibility, and / or executive function, as evidenced, for example, by TMT-A of greater than 30 seconds, or greater than 45 seconds, or greater than 60 seconds, and / or TMT-B of greater than 70 seconds, or greater than 90 seconds, or greater than 120 seconds, or greater than 150 seconds, or greater than 180 seconds, and / or TMT-B minus TMT-A of greater than 40 seconds, or greater than 60 seconds, or greater than 90 seconds, or greater than 120 seconds, Method 2.22. 2.24. The subject has Gaucher disease type 3, method 2 or any of 2.1-2.23; 2.25. The subject has Niemann-Pick disease type C, method 2 or any of 2.1-2.24; 2.26. Subjects are diagnosed with GM2-gangliosidosis (e.g., Tay-Sachs disease, Sandoz disease) Hoff's disease, or GM2 gangliosidosis AB variant), either method 2 or 2.1–2.24; 2.27. The subject has been diagnosed with a mutation in the gene GBA1, according to any of methods 2 or 2.1 to 2.24; 2.28. The subject has been diagnosed with a mutation in the genes NPC1 and / or NPC2, according to any of methods 2 or 2.1-2.24; 2.29. Any of Methods 2 or 2.1-2.24, wherein the subject has been diagnosed with a mutation in the gene HEXA (encoding hexosaminidase A) and / or a mutation in the gene HEXB (encoding hexosaminidase B) and / or a mutation in the gene GM2A (encoding GM2 ganglioside activator protein); 2.30. Subjects have been diagnosed with Parkinson's disease, either Method 2 or 2.1-2.29; 2.31. Any of Methods 2 or 2.1-2.30, wherein the subject is undergoing concurrent treatment with an enzyme replacement therapy (ERT), e.g., with glucocerebrosidase (e.g., imiglucerase, velaglucerase, or taliglucerase), and optionally, each such enzyme is a recombinant enzyme; 2.32. Method 2.31, wherein the subject is undergoing concurrent treatment with one or more of imiglucerase, velaglucerase (e.g., velaglucerase alfa), and taliglucerase (e.g., taliglucerase alfa); 2.33. The subject is receiving concurrent treatment with imiglucerase, Method 2.32; 2.34. The subject is receiving concurrent treatment with imiglucerase (1 unit of imiglucerase is the amount of enzyme that catalyzes the hydrolysis of 1 micromole of the synthetic substrate p-nitrophenyl-β-D-glucopyranoside per minute at 37°C) at a dosage of 2.5 units / kg body weight to 80 units / kg body weight every 1 to 3 weeks, e.g., 40 to 60 units / kg body weight every 2 weeks, Method 2.33; 2.35. Method 2.34, wherein the subject's dosage of imiglucerase in each administration (e.g., every 1 to 3 weeks, e.g., every 2 weeks) is administered as an intravenous (IV) infusion over 1 to 3 hours (e.g., 1 to 2 hours); 2.36. Any of Methods 2 or 2.1 through 2.35, wherein the subject has been administered an enzyme replacement therapy (e.g., imiglucerase, velaglucerase, and / or taliglucerase) prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 2.37. Method 2.36, wherein the subject has been administered an imiglucerase treatment for at least 6 months, e.g., at least 12 months (1 year), or at least 18 months, or at least 2 years, or at least 3 years, prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75). 2.38. Method 2.36 or 2.37, wherein the subject has been receiving imiglucerase therapy at a stable dose for at least 6 months prior to initiating therapy with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 2.39. Any of Methods 2 or 2.1-2.38, further comprising the step of transitioning the subject from an ERT therapeutic agent (e.g., imiglucerase, velaglucerase, or taliglucerase) to treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 2.40. The subject's hemoglobin level is at least 11 g / dL for women and at least 12 g / dL for men, using Method 2 or any of Methods 2.1 through 2.39. Either; 2.41. The subject's platelet count is at least 100,000 per cubic millimeter, as described in Method 2 or any of Methods 2.1 through 2.40; 2.42. Subject has a spleen volume less than 10 times normal (10 MN) and / or a liver volume less than 1.5 MN, either Method 2 or 2.1-2.41; 2.43. The subject has been diagnosed with a co-occurring dementia, e.g., Alzheimer's disease or Parkinson's disease, according to Method 2, or any of Methods 2.1-2.42; 2.44. Any of Methods 2 or 2.1-2.43, wherein the subject is at least 18 years of age (e.g., 18-30 years of age) at the start of treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 2.45. The subject's glucosylceramide (GL1) concentration is 4.4 to 11.1 ng / mL in cerebrospinal fluid (CSF) and 4.9 to 8.3 μg / mL in plasma, according to any of methods 2 or 2.1 to 2.44; 2.46. The subject's glucosylsphingosine (lyso-GL1) concentration is 20.1 to 67.6 pg / mL in CSF and 8.8 to 159.0 ng / mL in plasma, either in Methods 2 or 2.1 to 2.45; 2.47 any of Methods 2 or 2.1-2.46, administering to the subject a daily dose of a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75) selected from about 1 mg to about 150 mg, e.g., 5 to 50 mg, or 10 to 40 mg, or 10 to 30 mg, or 10 to 20 mg, or 20 to 30 mg, or 30 to 40 mg, or 40 to 50 mg, or 5 to 25 mg, or 20 to 50 mg, or 5 to 15 mg, or 15 to 30 mg, or about 15 mg, or 2, 5, 15, 25, 50, 100, or 150 mg; 2.48. Any of Methods 2 or 2.1-2.47, wherein the subject is a human adult patient, e.g., between 18 and 80 years of age, e.g., between 18 and 60 years of age, or between 18 and 40 years of age, or between 18 and 30 years of age, or between 18 and 25 years of age; 2.49. Any of Methods 2 or 2.1-2.47, wherein the subject is a human pediatric patient, e.g., 0 to 18 years of age, e.g., 1 to 15 years of age, or 1 to 5 years of age, or 5 to 10 years of age, or 10 to 15 years of age, or 10 to 18 years of age; 2.50. Any of Methods 2 or 2.1 through 2.49, which is effective to reduce the SARA ataxia scale by at least 0.5, e.g., to reduce the SARA score by at least 1, or at least 2, or at least 3, or at least 5, or at least 10; or the method is effective to reduce the SARA score between 0.00 and 3.00, or between 0.00 and 2.00, or between 0.00 and 1.50, or between 0.00 and 1.00, or between 0.00 and 0.50. 2.51. Effective in improving cognitive performance or reducing a cognitive deficit, e.g., as measured by a reduction in the time it takes to complete the Trail Making Test (TMT), TMT-A and / or TMT-B, a reduction in the difference between TMT-A and TMT-B times (TMT-A-TMT-B), e.g., by at least a 10%, or at least a 20%, or at least a 30%, or at least a 40%, or at least a 50% reduction (e.g., a 5-20% reduction in TMT-A and / or a 25-30% reduction in TMT-B and / or a 25-30% reduction in [TMT-A-TMT-B]), in any of Methods 2 or 2.1-2.50; 2.52. Any of Methods 2 or 2.1-2.51, wherein the method results in at least a 30%, e.g., at least a 40%, at least a 50%, at least a 60%, or at least a 70% decrease in glucosylceramide levels in the CSF and / or in the plasma after 6 months of treatment; 2.53. Any of Methods 2 or 2.1-2.52, wherein the method results in an increase in glucosylsphingosine concentration in the CSF and / or in the plasma of at least 30%, e.g., at least 40%, at least 50%, at least 60%, or at least 70%, after 6 months of treatment; 2.54. Any of Methods 2 or 2.1 through 2.53, wherein the method results in statistically or clinically unchanged modified Severity Scoring Tool (mSST) values ​​for neurological disease after 6 months of treatment; 2.55. Any of Methods 2 or 2.1-2.54, wherein the method results in increased blood flow in the brain (e.g., in one or more of the frontal lobe, occipital lobe, parietal lobe, or temporal lobe), e.g., as shown by fMRI imaging; 2.56. Any of Methods 2 or 2.1-2.54, wherein the method results in increased nodal connectivity in the brain (e.g., between the posterior and anterior sides of the brain, and / or between occipito-parietal structures and frontal, temporal, and / or limbic structures), e.g., as shown by fMRI imaging; 2.57. Any of Methods 2 or 2.1-2.56, where the method results in enhanced connectivity in brain regions associated with executive function; 2.58. Either Method 2 or 2.1-2.57, where the method results in a resting-state functional network with improved connectivity between the default mode and medial frontal networks; 2.59. Either Method 2 or 2.1–2.58, which results in enhanced connectivity between RSNs 1, 2, and 3 (perceptual-visual, cognitive-linguistic-orthographic, cognitive-spatial) and RSNs 6, 7, and 8 (sensorimotor, auditory, and executive control); 2.60. Method 2, or any of 2.1 to 2.59, wherein a compound according to Formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or any of Compounds 1 or 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a parenteral or non-parenteral route; 2.61. The route of administration is oral (enteral), Method 2.60; 2.62. Method 2.60, in which the route of administration is parenteral, e.g., by injection, e.g., by intravenous injection; 2.63. Method 2, or any of 2.1 to 2.62, wherein a compound according to formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or any of compounds 1 or 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by local administration, e.g., topical administration; 2.64. Method 2, or any of 2.1 through 2.63, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate or quinuclidin-3-yl(2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate; 2.65. The compound dosage is 15 mg / day orally, Method 2.64; 2.66. The compound dosage is 15 mg / day in a single oral dose, Method 2.65; 2.67. Method 2, or any of 2.1-2.66, wherein the subject is administered a single daily dose of 5 mg, 10 mg, 15 mg, or 20 mg of a compound, e.g., (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate.

[0094] Methods according to Method 2 and subsequent methods may be beneficial for subjects who have been diagnosed with a lysosomal storage disease, e.g., Gaucher Type 3 or Niemann-Pick Type C, but who have not yet experienced the cognitive and / or ataxic symptoms associated with the condition. Methods according to Method 2 and subsequent methods may be beneficial for subjects who have been diagnosed with a lysosomal storage disease, e.g., Gaucher Type 3 or Niemann-Pick Type C, but who have not yet experienced the cognitive and / or ataxic symptoms associated with the condition, e.g., due to a mutation in the subject or in the subject's family known to cause such a disease. It may also be beneficial for subjects at risk of developing lysosomal storage disease, such as Gaucher type 3 or Niemann-Pick type C.Therefore, in some embodiments of the methods described herein, the subject has been diagnosed as being at risk of developing the disease or disorder, and the method prevents or delays the onset and / or development of cognitive and / or ataxic symptoms of the disease or disorder in the subject.In some embodiments, the subject has been diagnosed as being at risk of developing the disease or disorder because they have a mutation in the gene described herein.

[0095] In a third aspect, the present invention provides a method for increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject, such as a subject in need thereof, the method comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compounds 1 or any of 1.1-1.75 (Method 3). Also provided are quinuclidine compounds described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compounds 1 or any of 1.1-1.75, for use in increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject in need thereof, e.g., for use in Method 3 or 3.1-3.65. Further provided is the use of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compound 1 or any of 1.1 to 1.75, in the manufacture of a medicament for increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject in need thereof, e.g., in the manufacture of a medicament for use in Method 3 or 3.1-3.65.

[0096] In certain further embodiments of Method 3, the present disclosure provides: 3.1. Method 3, comprising administering to a subject an effective amount of a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75; 3.2. Method 3, comprising administering to a subject an effective amount of Compound 1 or any one or more of Compounds 1.1-1.75; 3.3. Method 3, or any of Methods 3.1-3.2, comprising administering to a subject an effective amount of a pharmaceutical composition comprising a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75; 3.4. Any of Methods 3 or 3.1-3.3, comprising administering to a subject an effective amount of a pharmaceutical composition comprising Compound 1 or any one or more of Compounds 1.1-1.75; 3.5. Method 3.3 or 3.4, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient described herein; 3.6. Any of Methods 3 or 3.1-3.5, comprising administering a pharmaceutical dosage form comprising an effective amount of a compound or an effective amount of a pharmaceutical composition; 3.7. Method 3.6, wherein the dosage form is an oral dosage form (e.g., a pill, capsule, caplet, tablet, dragee, powder, granules, film, lozenge, or liquid); 3.8. Method 3.7, wherein the dosage form is a chewable tablet; 3.9. The dosage form is a parenteral dosage form (e.g., when the pharmaceutical composition is formulated for injection), Method 3.6; 3.10. Method 3.9, in which the injection is intravenous, intramuscular, intrathecal, or subcutaneous, and optionally is a sterile injection; 3.11. Method 3.6, wherein the dosage form is a topical or rectal dosage form; 3.12. Method 3.6, wherein the dosage form is an intranasal dosage form (e.g., an aerosol); 3.13. Any of Methods 3 or 3.1-3.12, further comprising simultaneously administering a second active agent, e.g., a second compound described herein that can reduce glycosylceramide levels in a patient in need thereof; 3.14. Method 3.13, wherein the second active agent is administered in the same pharmaceutical composition or dosage form as the quinuclidine compound; 3.15. Methods 3.13 or 3.14, wherein the second active agent is a GCS inhibitor (e.g., miglustat, or eliglustat); 3.16. Method 3 or any of 3.1-3.15, wherein the subject is a mammal; 3.17. Method 3.16, wherein the subject is a primate; 3.18. Method 3.17, wherein the subject is a human; 3.19. Method 3, or any of 3.1-3.18, wherein the subject has a symptom selected from ataxia, e.g., gait instability, asthenia, insynergia, slowed reaction time, dyschronometry, dysarthria, dysphagia, hypotonia, dysmetria, hypometria, hypermetria, dyspraxia of antagonistic movements, speech disorder, vocal tremor, ataxic breathing, postural instability, and combinations thereof, e.g., wherein the predominant ataxic deficit is gait instability; 3.20. Subjects will complete the Scale for Assessment and Rating of Ataxia at the start of treatment with the method. Method 3.19, having a baseline ataxia of at least 0.5 on the State of Ataxia (SARA) scale, e.g., a baseline SARA score of at least 1, or at least 2, or at least 3, or at least 4, or at least 5, or at least 10, or at least 20; 3.21. The subject has cognitive impairment (e.g., dementia), Method 3, or any of Methods 3.1-3.20; 3.22. Cognitive impairment is dementia, Method 3.21; 3.23. Dementia is characterized by signs of deficits in visual search speed, processing scanning speed, mental flexibility, and / or executive function, as evidenced, for example, by TMT-A of greater than 30 seconds, or greater than 45 seconds, or greater than 60 seconds, and / or TMT-B of greater than 70 seconds, or greater than 90 seconds, or greater than 120 seconds, or greater than 150 seconds, or greater than 180 seconds, and / or TMT-B minus TMT-A of greater than 40 seconds, or greater than 60 seconds, or greater than 90 seconds, or greater than 120 seconds, Method 3.22. 3.24. The subject has Gaucher disease type 3, method 3 or any of methods 3.1-3.23; 3.25. The subject has Niemann-Pick disease type C, and any of Methods 3 or 3.1 through 3.24; 3.26. The subject has GM2-gangliosidosis (e.g., Tay-Sachs disease, Sandhoff disease, or GM2 gangliosidosis AB variant), according to any of Methods 3 or 3.1-3.24; 3.27. The subject has been diagnosed with a mutation in the gene GBA1, according to any of methods 3 or 3.1 through 3.24; 3.28. The subject has been diagnosed with a mutation in the genes NPC1 and / or NPC2, according to any of Methods 3 or 3.1-3.24; 3.29. Any of Methods 3 or 3.1-3.24, wherein the subject has been diagnosed with a mutation in the gene HEXA (encoding hexosaminidase A) and / or a mutation in the gene HEXB (encoding hexosaminidase B) and / or a mutation in the gene GM2A (encoding GM2 ganglioside activator protein); 3.30. The subject has been diagnosed with Alzheimer's disease or Parkinson's disease, Method 3, or any of Methods 3.1 through 3.29; 3.31. Any of Methods 3 or 3.1-3.30, wherein the subject is undergoing concurrent treatment with an enzyme replacement therapy (ERT), e.g., with glucocerebrosidase (e.g., imiglucerase, velaglucerase, or taliglucerase), and optionally, each such enzyme is a recombinant enzyme; 3.32. Method 3.31, wherein the subject is undergoing concurrent treatment with one or more of imiglucerase, velaglucerase (e.g., velaglucerase alfa), and taliglucerase (e.g., taliglucerase alfa); 3.33. The subject is receiving concurrent treatment with imiglucerase, Method 3.32; 3.34. The subject is receiving concurrent treatment with imiglucerase at a dosage of 2.5 units / kg body weight to 80 units / kg body weight every 1 to 3 weeks, e.g., 40 to 60 units / kg body weight every 2 weeks (1 unit of imiglucerase is the amount of enzyme that catalyzes the hydrolysis of 1 micromole of the synthetic substrate p-nitrophenyl-β-D-glucopyranoside per minute at 37°C), Method 3.33; 3.35. Method 3.34, wherein the subject's dosage of imiglucerase in each administration (e.g., every 1 to 3 weeks, e.g., every 2 weeks) is administered as an intravenous (IV) infusion over 1 to 3 hours (e.g., 1 to 2 hours); 3.36. Any of Methods 3 or 3.1 through 3.35, wherein the subject is administered an enzyme replacement therapy (e.g., imiglucerase, velaglucerase, and / or taliglucerase) prior to initiating treatment with a compound according to Formula (I) (or any of (II) through (XII), (Ia) through (XIIa), or (Ib) through (XIIb), or any of Compounds 1 or 1.1 through 1.75); 3.37. Method 3.36, wherein the subject has been administered an imiglucerase treatment for at least 6 months, e.g., at least 12 months (1 year), or at least 18 months, or at least 2 years, or at least 3 years, prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75). 3.38. Method 3.36 or 3.37, wherein the subject has been receiving imiglucerase therapy at a stable dose for at least 6 months prior to initiating therapy with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 3.39. Any of Methods 3 or 3.1-3.38, further comprising the step of transitioning the subject from an ERT therapeutic agent (e.g., imiglucerase, velaglucerase, or taliglucerase) to treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 3.40. The subject's hemoglobin level is at least 11 g / dL for women and at least 12 g / dL for men, according to Method 3 or any of Methods 3.1 through 3.39; 3.41. The subject's platelet count is at least 100,000 per cubic millimeter, according to Method 3 or any of Methods 3.1 through 3.40; 3.42. Subject has a spleen volume less than 10 times normal (10 MN) and / or a liver volume less than 1.5 MN, Method 3, or any of 3.1-3.41; 3.43. The subject has been diagnosed with a co-occurring dementia, e.g., Alzheimer's disease or Parkinson's disease, according to Method 3, or any of Methods 3.1-3.42; 3.44. Any of Methods 3 or 3.1-3.43, wherein the subject is at least 18 years of age (e.g., 18-30 years of age) at the start of treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 3.45. The subject's glucosylceramide (GL1) concentration is 4.4 to 11.1 ng / mL in cerebrospinal fluid (CSF) and 4.9 to 8.3 μg / mL in plasma. Law 3 or any of 3.1-3.44; 3.46. The subject's glucosylsphingosine (lyso-GL1) concentration is 20.1 to 67.6 pg / mL in CSF and 8.8 to 159.0 ng / mL in plasma, according to any of Methods 3 or 3.1 to 3.45; 3.47. Any of Methods 3 or 3.1-3.46, administering to the subject a daily dose of a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75) selected from about 1 mg to about 150 mg, e.g., 5 to 50 mg, or 10 to 40 mg, or 10 to 30 mg, or 10 to 20 mg, or 20 to 30 mg, or 30 to 40 mg, or 40 to 50 mg, or 5 to 25 mg, or 20 to 50 mg, or 5 to 15 mg, or 15 to 30 mg, or about 15 mg, or 2, 5, 15, 25, 50, 100, or 150 mg; 3.48. Method 3 or any of 3.1-3.47, wherein the subject is a human adult patient, e.g., between 18 and 80 years of age, e.g., between 18 and 60 years of age, or between 18 and 40 years of age, or between 18 and 30 years of age, or between 18 and 25 years of age; 3.49. Method 3 or any of 3.1-3.47, wherein the subject is a human pediatric patient, e.g., 0 to 18 years of age, e.g., 1 to 15 years of age, or 1 to 5 years of age, or 5 to 10 years of age, or 10 to 15 years of age, or 10 to 18 years of age; 3.50. Any of Methods 3 or 3.1 through 3.49, wherein the method results in an increase in brain tissue volume, or prevention or delay of brain tissue volume loss, in one or more brain regions selected from the right nucleus accumbens, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left pericalcarine, right amygdala, left cuneus, and left lingual gyrus, as measured, e.g., using volumetric magnetic resonance imaging (vMRI); 3.51. Any of Methods 3 or 3.1 through 3.50, wherein the method results in an increase in brain tissue volume in one or more brain regions associated with executive function; 3.52. Method 3.50 or 3.51, wherein an increase in brain tissue volume in one or more brain regions is accompanied by enhanced neuronal connectivity within one or more brain regions, as shown, for example, using functional magnetic resonance imaging (fMRI); 3.53. Any of Method 3 or 3.1 through 3.52, wherein the method results in an increase in total brain tissue volume; 3.54. The increase in brain tissue volume is at least 5 mm in any one or more brain regions. 3 , e.g., at least 10 mm in any one or more brain regions. 3 , at least 15mm 3 , at least 20 mm 3 , at least 30mm 3 , at least 50mm 3 , at least 70mm 3 , or at least 90mm 3 , and / or up to 100 mm in any one or more brain regions 3 , or up to 150mm 3 , either Method 3 or 3.1 to 3.53; 3.55. The increase in total brain tissue volume is at least 5 mm 3 , e.g., at least 30 mm 3 , at least 60mm 3 , at least 90mm 3 , at least 120mm 3 , at least 150mm 3 , at least 200mm 3 , or at least 250mm 3 , and / or total brain volume up to 400 mm 3 , or up to 500mm 3 , either Method 3 or 3.1 to 3.54; 3.56. Any of Methods 3 or 3.1-3.55, wherein the increase in brain tissue volume is at least 0.1%, e.g., 0.1% to 10.0%, e.g., at least 0.50%, 0.75%, 1.0%, 2.0%, or 5.0%, in any one or more brain regions, compared to the initial brain tissue volume; 3.57. Method 3 or any of 3.1-3.56, wherein the increase in total brain tissue volume is at least 0.05%, e.g., 0.05% to 0.30%, e.g., at least 0.10%, 0.15%, 0.20%, or 0.25%, compared to the initial total brain tissue volume; 3.58. Compounds according to formula (I) (or (II) to (XII), (Ia) to (XII) Method 3, or any of 3.1 to 3.57, wherein compound 1 or any of 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a parenteral or non-parenteral route; 3.59. The route of administration is oral (enteral), Method 3.58; 3.60. Method 3.58, in which the route of administration is parenteral, e.g., by injection, e.g., intravenous injection; 3.61. Method 3, or any of 3.1 to 3.57, wherein a compound according to Formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or any of Compounds 1 or 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by local administration, e.g., topical administration; 3.62. Method 3, or any of 3.1 through 3.61, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate or quinuclidin-3-yl(2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate; 3.63. The compound dosage is 15 mg / day orally, Method 3.62; 3.64. The compound dosage is 15 mg / day in a single oral dose, Method 3.63; 3.65. Method 3, or any of 3.1-3.61, wherein the subject is administered a single daily dose of 5 mg, 10 mg, 15 mg, or 20 mg of a compound, e.g., (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate.

[0097] Methods according to Method 3 and beyond may be beneficial for subjects who have been diagnosed with a lysosomal storage disease, such as Gaucher Type 3 or Niemann-Pick Type C, but who have not yet experienced the cognitive and / or ataxic symptoms associated with the condition. Methods according to Method 3 and beyond may also be beneficial for subjects who are at risk of developing a lysosomal storage disease, such as Gaucher Type 3 or Niemann-Pick Type C, for example, due to a mutation in the subject or the subject's family known to cause such a disease. Thus, in some embodiments of the methods described herein, the subject has been diagnosed as being at risk for developing the disease or disorder, and the method prevents or delays the onset and / or development of cognitive and / or ataxic symptoms of the disease or disorder in the subject. In some embodiments, the subject has been diagnosed as being at risk for developing the disease or disorder because they have a mutation in a gene described herein.

[0098] The methods according to Method 3 and subsequent methods aim to increase brain tissue volume, and there are no particular limitations on possible brain regions that may be beneficially increased in terms of tissue volume. Examples of relevant brain regions include the right putamen, right postcentral lobe, right amygdala, right lingual gyrus, left cuneus, left lingual gyrus, right superior temporal lobe, right lateral orbitofrontal lobe, left calcarine sulcus lobe, left lateral temporal lobe, right temporal pole, right nucleus accumbens, left putamen, left entorhinal cortex, right globus pallidus, left nucleus accumbens, left temporal pole, right entorhinal cortex, left caudate nucleus, right frontal pole, right operculum, right lateral temporal lobe, right hippocampus, left paracentral lobule, left superior parietal lobe, left fusiform gyrus, left parietal of the STS, right paracentral lobe, right medial temporal lobe, left caudate-middle frontal lobe, right rostral anterior cingulate lobe, left paratriangular lobe, left precentral lobe, right orbital lobe, left middle temporal lobe, and left isthmus of the cingulate gyrus. lobe), right caudate-middle frontal lobe, entire right temporal lobe, left inferior temporal lobe, right lateral parietal lobe, right lateral occipital lobe, left supramarginal gyrus, entire left temporal lobe, right rostral middle frontal lobe, left rostral middle frontal lobe, left medial temporal lobe, and left superior frontal lobe, or any combination thereof. In at least some embodiments, specific brain regions that may show the greatest relative volume increase as a result of methods according to Method 3 or later include the right nucleus accumbens, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine sulcus, right amygdala, left cuneus, and left lingual gyrus.

[0099] Monitoring and evaluation methods In a fourth aspect, the present invention provides a method (Method 4) for monitoring the progression or regression of a neurological disorder associated with a lysosomal storage disease in a subject, wherein the subject is receiving treatment comprising administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or Compounds 1 or 1.1-1.75; the method comprises measuring the subject's brain tissue volume over a period of time during the course of treatment and assessing the degree of change in brain tissue volume over said period, e.g., using volumetric magnetic resonance imaging (vMRI), and optionally further comprising initiating or adjusting the subject's treatment by administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or Compounds 1 or 1.1-1.75. As will be appreciated, to assess the extent of brain tissue volume change as a result of treatment, measurements of brain tissue volume can be performed, for example, at the beginning of the above-described treatment, or shortly thereafter (e.g., 1-14 days), and at the end of a designated treatment period, or intermittently / periodically over the course of ongoing treatment (e.g., weekly, monthly, every 2, 3, 4, 6, 9, 12 months, etc.) to assess and reassess the extent of brain volume change over the course of treatment. By accumulating comparative results regarding the subject's condition over the period in which the subject is receiving treatment, greater accuracy can be achieved in determining the subject's condition and progression / regression of disease symptoms.

[0100] In certain further embodiments of Method 4, the present disclosure provides: 4.1. Method 4, wherein the treatment comprises administering to the subject an effective amount of a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75; 4.2. Method 4, wherein the treatment comprises administering to the subject an effective amount of Compound 1 or any one or more of Compounds 1.1-1.75; 4.3. Method 4, or any of 4.1-4.2, wherein the treatment comprises administering to the subject an effective amount of a pharmaceutical composition comprising a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75; 4.4. Any of Methods 4 or 4.1-4.2, wherein the treatment comprises administering to the subject an effective amount of a pharmaceutical composition comprising Compound 1 or any one or more of Compounds 1.1-1.75; 4.5. Method 4.3 or 4.4, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient described herein; 4.6. Any of Methods 4 or 4.1-4.5, wherein the treatment comprises administering a pharmaceutical dosage form containing an effective amount of the compound or an effective amount of the pharmaceutical composition; 4.7. Method 4.6, wherein the dosage form is an oral dosage form (e.g., a pill, capsule, caplet, tablet, dragee, powder, granules, film, lozenge, or liquid); 4.8. Method 4.7, wherein the dosage form is a chewable tablet; 4.9. The dosage form is a parenteral dosage form (e.g., when the pharmaceutical composition is formulated for injection), Method 4.6; 4.10. Method 4.9, in which the injection is intravenous, intramuscular, intrathecal, or subcutaneous, and optionally is a sterile injection; 4.11. Method 4.6, wherein the dosage form is a topical or rectal dosage form; 4.12. Method 4.6, wherein the dosage form is an intranasal dosage form (e.g., an aerosol); 4.13. The treatment is with a second active agent, e.g., a second compound described herein that can reduce glycosylceramide levels in a patient in need thereof. any of Methods 4 or 4.1 to 4.12, further comprising simultaneously administering 4.14. Method 4.13, wherein the second active agent is administered in the same pharmaceutical composition or dosage form as the quinuclidine compound; 4.15. Method 4.13 or 4.14, wherein the second active agent is a GCS inhibitor (e.g., miglustat or eliglustat); 4.16. Method 4 or any of 4.1-4.15, wherein the subject is a mammal; 4.17. Method 4.16, wherein the subject is a primate; 4.18. Method 4.17, wherein the subject is a human; 4.19. Method 4, or any of 4.1-4.18, wherein the subject has a symptom selected from ataxia, e.g., gait instability, asthenia, insynergia, slowed reaction time, dyschronometry, dysarthria, dysphagia, hypotonia, dysmetria, hypometria, hypermetria, dyspraxia of antagonistic movements, speech disorder, vocal tremor, ataxic breathing, postural instability, and combinations thereof, e.g., wherein the predominant ataxic deficit is gait instability; 4.20. Subjects will complete the Scale for Assessment and Rating of Ataxia at the start of treatment with the method. Method 4.19, having a baseline ataxia of at least 0.5 on the State of Ataxia (SARA) scale, e.g., a baseline SARA score of at least 1, or at least 2, or at least 3, or at least 4, or at least 5, or at least 10, or at least 20; 4.21. The subject has cognitive impairment (e.g., dementia), Method 4, or any of Methods 4.1-4.20; 4.22. Cognitive impairment is dementia, Method 4.21; 4.23. Dementia is characterized by signs of deficits in visual search speed, scanning processing speed, mental flexibility, and / or executive function, as evidenced, for example, by TMT-A of greater than 30 seconds, or greater than 45 seconds, or greater than 60 seconds, and / or TMT-B of greater than 70 seconds, or greater than 90 seconds, or greater than 120 seconds, or greater than 150 seconds, or greater than 180 seconds, and / or TMT-B minus TMT-A of greater than 40 seconds, or greater than 60 seconds, or greater than 90 seconds, or greater than 120 seconds, Method 4.22; 4.24. The subject has Gaucher disease type 3, method 4 or any of methods 4.1 through 4.23; 4.25. The subject has Niemann-Pick disease type C, and any of Methods 4 or 4.1 through 4.24; 4.26. The subject has GM2-gangliosidosis (e.g., Tay-Sachs disease, Sandhoff disease, or GM2 gangliosidosis AB variant), according to any of Methods 4 or 4.1-4.24; 4.27. The subject has been diagnosed with a mutation in the gene GBA1, according to any of methods 5 or 4.1-4.24; 4.28. Method 5 or any of 4.1-4.24, wherein the subject has been diagnosed with a mutation in the genes NPC1 and / or NPC2; 4.29. Any of Methods 5 or 4.1-4.24, wherein the subject has been diagnosed with a mutation in the gene HEXA (encoding hexosaminidase A) and / or a mutation in the gene HEXB (encoding hexosaminidase B) and / or a mutation in the gene GM2A (encoding GM2 ganglioside activator protein); 4.30. The subject has been diagnosed with Alzheimer's disease or Parkinson's disease, according to any of Methods 4 or 4.1 through 4.29; 4.31. Any of Methods 4 or 4.1-4.30, wherein the subject is undergoing concurrent treatment with an enzyme replacement therapy (ERT), e.g., with a glucocerebrosidase (e.g., imiglucerase, velaglucerase, or taliglucerase), optionally each such enzyme being a recombinant enzyme; 4.32. Method 4.31, wherein the subject is undergoing concurrent treatment with one or more of imiglucerase, velaglucerase (e.g., velaglucerase alfa), and taliglucerase (e.g., taliglucerase alfa); 4.33. The subject is receiving concurrent treatment with imiglucerase, Method 4.32; 4.34. The subject is receiving concurrent treatment with imiglucerase (1 unit of imiglucerase is the amount of enzyme that catalyzes the hydrolysis of 1 micromole of the synthetic substrate p-nitrophenyl-β-D-glucopyranoside per minute at 37°C) at a dosage of 2.5 units / kg body weight to 80 units / kg body weight every 1 to 3 weeks, e.g., 40 to 60 units / kg body weight every 2 weeks, Method 4.33; 4.35. Method 4.34, wherein the subject's dosage of imiglucerase in each administration (e.g., every 1 to 3 weeks, e.g., every 2 weeks) is administered as an intravenous (IV) infusion over 1 to 3 hours (e.g., 1 to 2 hours); 4.36. Any of Methods 4 or 4.1 to 4.35, wherein the subject has been administered an enzyme replacement therapy (e.g., imiglucerase, velaglucerase, and / or taliglucerase) prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1 to 1.75); 4.37. Method 4.36, wherein the subject has been administered an imiglucerase treatment for at least 6 months, e.g., at least 12 months (1 year), or at least 18 months, or at least 2 years, or at least 3 years, prior to initiating treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75). 4.38. Method 4.36 or 4.37, wherein the subject has been receiving imiglucerase therapy at a stable dose for at least 6 months prior to initiating therapy with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 4.39. Any of Methods 4 or 4.1-4.38, further comprising the step of transitioning the subject from an ERT therapeutic agent (e.g., imiglucerase, velaglucerase, or taliglucerase) to treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 4.40. The subject's hemoglobin level is at least 11 g / dL for women and at least 12 g / dL for men, according to any of Methods 4 or 4.1 through 4.39; 4.41. The subject's platelet count is at least 100,000 per cubic millimeter, according to Method 4 or any of Methods 4.1 through 4.40; 4.42. Subject has a spleen volume less than 10 times normal (10 MN) and / or a liver volume less than 1.5 MN, either Method 4 or 4.1-4.41; 4.43. The subject has been diagnosed with a co-occurring dementia, e.g., Alzheimer's disease or Parkinson's disease, according to any of Methods 4 or 4.1-4.42; 4.44. Any of Methods 4 or 4.1-4.43, wherein the subject is at least 18 years of age (e.g., 18-30 years of age) at the start of treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 4.45. Any of Methods 4 or 4.1 through 4.44, wherein the subject's glucosylceramide (GL1) concentration is 4.4 to 11.1 ng / mL in cerebrospinal fluid (CSF) and 4.9 to 8.3 μg / mL in plasma; 4.46. The subject's glucosylsphingosine (lyso-GL1) concentration is 20.1 to 67.6 pg / mL in CSF and 8.8 to 159.0 ng / mL in plasma, according to any of Methods 4 or 4.1 to 4.45; 4.47. Any of Methods 4 or 4.1-4.46, administering to the subject a daily dose of a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75) selected from about 1 mg to about 150 mg, e.g., 5 to 50 mg, or 10 to 40 mg, or 10 to 30 mg, or 10 to 20 mg, or 20 to 30 mg, or 30 to 40 mg, or 40 to 50 mg, or 5 to 25 mg, or 20 to 50 mg, or 5 to 15 mg, or 15 to 30 mg, or about 15 mg, or 2, 5, 15, 25, 50, 100, or 150 mg; 4.48. Any of Methods 4 or 4.1-4.47, wherein the subject is a human adult patient, e.g., between 18 and 80 years of age, e.g., between 18 and 60 years of age, or between 18 and 40 years of age, or between 18 and 30 years of age, or between 18 and 25 years of age; 4.49. Any of Methods 4 or 4.1-4.47, wherein the subject is a human pediatric patient, e.g., 0 to 18 years of age, e.g., 1 to 15 years of age, or 1 to 5 years of age, or 5 to 10 years of age, or 10 to 15 years of age, or 10 to 18 years of age; 4.50. Method 4, or any of 4.1-4.49, wherein the period during which the subject undergoes a course of treatment and brain volume is monitored is 3 to 24 months, e.g., 3 to 12 months, 3 to 6 months, 6 to 24 months, 6 to 18 months, 6 to 12 months, 12 to 24 months, or 12 to 18 months; 4.51. Method 4, or any of 4.1-4.50, wherein the measurement of the subject's brain tissue volume over said period is by brain positron emission tomography (PET) or volumetric magnetic resonance imaging (vMRI). 4.52. Method 4.51, in which the subject's brain tissue volume is measured multiple times intermittently or periodically over the course of treatment, e.g., weekly, monthly, every 2, 3, 4, 6, 9, 12 months, etc.; 4.53. If there is a decrease or no increase in total brain volume observed over said period, the method further comprises modifying the treatment by increasing the dosage of a compound of Formula (I) (or any of (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75) administered to the subject during treatment, and reassessing the extent of change in brain tissue volume after a further period over the course of the modified treatment with increased dosage. 4.54. Method 4, or any of 4.1-4.53, wherein if there is a decrease or no increase in volume in three or more of the following brain regions observed over said period: right nucleus accumbens region, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine sulcus, right amygdala, left cuneus, and left lingual gyrus, the method further comprises modifying the treatment by increasing the dosage of a compound of Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75) administered to the subject during treatment, and re-evaluating the degree of change in brain tissue volume after a further period over the course of the modified treatment with the increased dosage. 4.55. Any of Methods 4 or 4.1 through 4.54, wherein the treatment results in an increase in brain tissue volume, or prevents or slows the loss of brain tissue volume, over a period of time in one or more brain regions selected from the right nucleus accumbens, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine sulcus, right amygdala, left cuneus, and left lingual gyrus; 4.56. Any of Methods 4 or 4.1-4.55, wherein the treatment results in an increase in brain volume in one or more brain regions associated with executive function; 4.57. Method 4.55 or 4.56, wherein an increase in brain volume in one or more brain regions is accompanied by enhanced neuronal connectivity within one or more brain regions, as demonstrated using functional magnetic resonance imaging (fMRI); 4.58. Method 4 or 4.1, wherein the treatment results in an increase in total brain tissue volume. Any of 4.57; 4.59. The increase in brain tissue volume is at least 5 mm in any one or more brain regions. 3 , e.g., at least 10 mm in any one or more brain regions. 3 , at least 15mm 3 , at least 20 mm 3 , at least 30mm 3 , at least 50mm 3 , at least 70mm 3 , or at least 90mm 3 , and / or up to 100 mm in any one or more brain regions 3 , or up to 150mm 3 Either method 4 or 4.55-4.58; 4.60 The increase in total brain tissue volume is at least 5 mm 3 , e.g., at least 30 mm 3 , at least 60mm 3 , at least 90mm 3 , at least 120mm 3 , at least 150mm 3 , at least 200mm 3 , or at least 250mm 3 , and / or total brain volume up to 400 mm 3 , or up to 500mm 3 Either method 4 or 4.55-4.59; 4.61. Any of Methods 4 or 4.55-4.60, wherein the increase in brain tissue volume is at least 0.1%, e.g., 0.1% to 10.0%, e.g., at least 0.50%, 0.75%, 1.0%, 2.0%, or 5.0%, in any one or more brain regions, compared to the initial brain tissue volume; 4.62. Any of methods 4 or 4.55-4.61, wherein the increase in total brain tissue volume is at least 0.05%, e.g., 0.05% to 0.30%, e.g., at least 0.10%, 0.15%, 0.20%, or 0.25%, compared to the initial brain tissue volume; 4.63. Method 4, or any of 4.1 to 4.62, wherein a compound according to Formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or any of Compounds 1 or 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a parenteral or non-parenteral route; 4.64. The route of administration is oral (enteral), Method 4.63; 4.65. Method 4.63, in which the route of administration is parenteral, e.g., by injection, e.g., by intravenous injection; 4.66. Method 4, or any of 4.1 to 4.63, wherein a compound according to Formula (I) (or any of (II) to (XII), (Ia) to (XIIa) or (Ib) to (XIIb), or any of Compounds 1 or 1.1 to 1.75), or a pharmaceutically acceptable salt or prodrug thereof, is administered by local administration, e.g., topical administration; 4.67. Method 4, or any of 4.1 through 4.67, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate or quinuclidin-3-yl(2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate; 4.68. The compound dosage is 15 mg / day orally, Method 4.63; 4.69. The compound dosage is 15 mg / day in a single oral dose, Method 4.63; 4.70. Method 4, or any of 4.1 through 4.67, wherein the subject is administered a single daily dose of 5 mg, 10 mg, 15 mg, or 20 mg of a compound, e.g., (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate.

[0101] In certain embodiments, methods according to method 4 and subsequent methods may be performed using volumetric magnetic resonance imaging (vMRI) as described herein. For example, analytical tools to assist in the evaluation of vMRI data, including tensor-based morphometry (TBM), as described herein, may be used.

[0102] In some embodiments, methods 4 and beyond instead involve treating or preventing a neurological disorder associated with a lysosomal storage disease in a subject (e.g., a patient) in need thereof. It will be understood that the present invention can be viewed as a method of administering to a subject a quinuclidine compound according to any one of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compounds 1 or 1.1-1.75, in an amount of 100 mg / kg or more, and that the method can be considered a method of administering to a subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any one of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or Compounds 1 or 1.1-1.75; and that the method includes measuring the subject's brain tissue volume over a period of time during the course of treatment, e.g., using vMRI to assess the extent of change in brain tissue volume over said period of time, and adjusting the parameters of the treatment regimen accordingly.

[0103] In a fifth aspect, the present invention provides a method (Method 5) for assessing the onset of a neurological disorder associated with a lysosomal storage disease in a subject at risk of developing said neurological disorder, said method comprising: a) measuring the subject's brain tissue volume (e.g., using vMRI) and comparing it with a reference standard to assess whether the brain tissue volume is lower than the reference standard; and b) identifying the onset of said neurological disorder if the brain tissue volume identified in step (a) is lower than the reference standard; and optionally further comprising: c) initiating treatment of the subject by administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compound 1, or any of 1.1-1.75. As will be appreciated, by measuring a subject's brain tissue volume and comparing it to a reference standard, it is possible to determine whether a subject who has a lysosomal storage disease and is at risk of developing an associated neurological disorder has reached a stage in disease progression where treatment with, for example, a compound of formula (I), may be particularly beneficial.

[0104] Reference standard brain tissue volumes (whole brain tissue volume and / or individual brain tissue region volumes) can be determined based on healthy (dementia-free) population data (obtained or available from public sources), allowing age- and sex-specific normative volumetric data to be used as a reference standard based on the subject's age and sex. This allows for meaningful comparisons to assess a subject's disease progression based on the extent to which the subject's brain tissue volume is lower than the reference standard in brain tissue regions and / or based on whole brain tissue volume, from which the onset of a neurological disorder associated with a lysosomal storage disease can be determined.

[0105] In certain further embodiments of Method 5, the present disclosure provides: 5.1. Method 5, wherein the subject has Gaucher disease type 3; 5.2. The subject has Niemann-Pick disease type C, Method 5; 5.3. Method 5, wherein the subject has GM2-gangliosidosis (e.g., Tay-Sachs disease, Sandhoff disease, or GM2 gangliosidosis AB variant); 5.4. The subject has been diagnosed with a mutation in the gene GBA1, according to any of methods 5 or 5.1-5.3; 5.5. Any of Methods 5 or 5.1-5.3, wherein the subject has been diagnosed with a mutation in the genes NPC1 and / or NPC2; 5.6. Method 5 or any of 5.1-5.3, wherein the subject has been diagnosed with a mutation in the gene HEXA (encoding hexosaminidase A) and / or a mutation in the gene HEXB (encoding hexosaminidase B) and / or a mutation in the gene GM2A (encoding GM2 ganglioside activator protein); 5.7. Method 5 or any of 5.1-5.6, wherein the subject has been diagnosed with Alzheimer's disease or Parkinson's disease; 5.8. Method 5, wherein the subject is undergoing treatment with an enzyme replacement therapy (ERT), e.g., with glucocerebrosidase (e.g., imiglucerase, velaglucerase, or taliglucerase), and optionally each such enzyme is a recombinant enzyme. or any of 5.1-5.7; 5.9. Method 5.8, wherein the subject is undergoing treatment with one or more of imiglucerase, velaglucerase (e.g., velaglucerase alfa), and taliglucerase (e.g., taliglucerase alfa); 5.10. Method 5.9, wherein the subject is undergoing concurrent treatment with imiglucerase; 5.11. Method 5.10, wherein the subject is receiving treatment with imiglucerase (1 unit of imiglucerase is the amount of enzyme that catalyzes the hydrolysis of 1 micromole of the synthetic substrate p-nitrophenyl-β-D-glucopyranoside per minute at 37°C) at a dosage of 2.5 units / kg body weight to 80 units / kg body weight every 1 to 3 weeks, e.g., 40 to 60 units / kg body weight every 2 weeks; 5.12. Method 5.11, wherein the subject's dosage of imiglucerase in each administration (e.g., every 1 to 3 weeks, e.g., every 2 weeks) is administered as an intravenous (IV) infusion over 1 to 3 hours (e.g., 1 to 2 hours); 5.13. Any of Methods 5 or 5.1 to 5.12, wherein the subject has been administered an enzyme replacement therapy (e.g., imiglucerase, velaglucerase, and / or taliglucerase) prior to initiating any treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 5.14. Method 5.13, wherein the subject has been administered an imiglucerase treatment for at least 6 months, e.g., at least 12 months (1 year), or at least 18 months, or at least 2 years, or at least 3 years, prior to initiating any treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75). 5.15. Method 5.13 or 5.14, wherein the subject has been receiving imiglucerase therapy at a stable dose for at least 6 months prior to initiating any therapy with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 5.16. Any of Methods 5 or 5.1-5.15 further comprising the step of transitioning the subject from an ERT therapeutic agent (e.g., imiglucerase, velaglucerase, or taliglucerase) to any treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 5.17. Method 5 or any of 5.1-5.16, wherein the subject's hemoglobin level is at least 11 g / dL for women and at least 12 g / dL for men; 5.18. Method 5 or any of 5.1 through 5.17, wherein the subject's platelet count is at least 100,000 per cubic millimeter; 5.19. Method 5, or any of 5.1-5.18, where the subject has a spleen volume less than 10 times normal (10 MN) and / or a liver volume less than 1.5 MN; 5.20. The subject has been diagnosed with a co-occurring dementia, e.g., Alzheimer's disease or Parkinson's disease, according to any of Methods 5 or 5.1-5.19; 5.21. Any of Methods 5 or 5.1-5.20, wherein the subject is at least 18 years of age (e.g., 18-30 years of age) at the start of treatment with a compound according to Formula (I) (or any of (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75); 5.22. Any of Methods 5 or 5.1-5.21, wherein the subject's glucosylceramide (GL1) concentration is 4.4-11.1 ng / mL in cerebrospinal fluid (CSF) and 4.9-8.3 μg / mL in plasma; 5.23. The subject's glucosylsphingosine (lyso-GL1) concentration is 20.1-67.6 pg / mL in CSF and 8.8-159.0 ng / mL in plasma. , either Method 5 or 5.1 to 5.22; 5.24. Any of Methods 5 or 5.1-5.23, wherein the subject is a human adult patient, e.g., between 18 and 80 years of age, e.g., between 18 and 60 years of age, or between 18 and 40 years of age, or between 18 and 30 years of age, or between 18 and 25 years of age; 5.25. Any of Methods 5 or 5.1-5.23, wherein the subject is a human pediatric patient, e.g., 0 to 18 years of age, e.g., 1 to 15 years of age, or 1 to 5 years of age, or 5 to 10 years of age, or 10 to 15 years of age, or 10 to 18 years of age; 5.26. Measurement of the subject's brain tissue volume is by brain positron emission tomography (PET) or volumetric magnetic resonance imaging (vMRI), Method 5, or any of 5.1 through 5.25. 5.27. The subject's brain tissue volume is found to be lower than the reference standard, method 5, or any of 5.1 through 5.26; 5.28. Method 5.28, wherein comparison with a reference standard indicates that the subject has lower brain tissue volume in one or more brain regions selected from the right nucleus accumbens, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine sulcus, right amygdala, left cuneus, and left lingual gyrus. 5.29. Method 5.27 or 5.28, wherein comparison to a reference standard indicates that the subject has lower brain tissue volume in one or more brain regions associated with executive function; 5.30. Any of methods 5.27-5.29, wherein comparison with a reference standard indicates that the subject has lower brain tissue volume in one or more brain regions assessed as having loss of neuronal connectivity, as shown, for example, using functional magnetic resonance imaging (fMRI); 5.31. Any of the methods of 5.27-5.30, wherein comparison with a reference standard indicates that the subject has a lower total brain tissue volume; 5.32. Any of methods 5.27-5.31, further comprising initiating treatment of the subject by administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa) or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75; 5.33. Method 5.32, further comprising initiating treatment of the subject by administering to the subject an effective amount of a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compound 1, or any of 1.1-1.75; 5.34. Method 5.32 or 5.33, further comprising initiating treatment of the subject by administering to the subject an effective amount of Compound 1 or any one or more of Compounds 1.1-1.75; 5.35. Methods 5.32-5.34, wherein the treatment comprises administering to the subject an effective amount of a pharmaceutical composition comprising a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75; 5.36. Methods 5.32-5.35, wherein the treatment comprises administering to the subject an effective amount of a pharmaceutical composition comprising Compound 1 or any one or more of Compounds 1.1-1.75; 5.37. Method 5.35 or 5.36, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient described herein; 5.38. Methods 5.32-5.37, further comprising initiating treatment of the subject by administering a pharmaceutical dosage form containing an effective amount of the compound or an effective amount of the pharmaceutical composition; 5.39. Method 5.38, wherein the dosage form is an oral dosage form (e.g., a pill, capsule, caplet, tablet, dragee, powder, granules, film, lozenge, or liquid); 5.40. Method 5.39, wherein the dosage form is a chewable tablet; 5.41. The dosage form is a parenteral dosage form (e.g., when the pharmaceutical composition is formulated for injection), Method 5.38; 5.42. The injection is intravenous, intramuscular, intrathecal, or subcutaneous, optionally a sterile injection, Method 5.41; 5.43. Method 5.38, wherein the dosage form is a topical or rectal dosage form; 5.44. Method 5.38, wherein the dosage form is an intranasal dosage form (e.g., an aerosol); 5.45. Methods 5.32-5.44, wherein the treatment further comprises simultaneously administering a second active agent, e.g., a second compound described herein capable of reducing glycosylceramide levels in a patient in need thereof; 5.46. Method 5.45, wherein the second active agent is administered in the same pharmaceutical composition or dosage form as the quinuclidine compound; 5.47. Method 5.45 or 5.46, wherein the second active agent is a GCS inhibitor (e.g., miglustat or eliglustat); 5.48. Any of methods 5.32-5.47, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate or quinuclidin-3-yl(2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate; 5.49. The compound dosage is 15 mg / day orally, Method 5.48; 5.50. The compound dosage is 15 mg / day in a single oral dose, Method 5.49; 5.51. Any of methods 5.32-5.48, wherein the subject is administered a single daily dose of 5 mg, 10 mg, 15 mg, or 20 mg of a compound, e.g., (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate; 5.52. Any of methods 5.32-5.51, wherein the subject's brain tissue volume is measured multiple times intermittently or periodically, e.g., weekly, monthly, every 2, 3, 4, 6, 9, 12 months, etc., after treatment with a compound according to Formula (I) is initiated, to assess changes in brain tissue volume; 5.53. Method 5 or any of 5.1 through 5.52, wherein the subject is a mammal; 5.54. Method 5.53, in which the subject is a primate; 5.55. The subject is a human, method 5.54;

[0106] It will be understood that in some embodiments, methods 5 onward may instead be considered methods of treating or preventing a neurological disorder associated with a lysosomal storage disease in a subject (e.g., a patient) in need thereof and at risk of developing said neurological disorder, comprising assessing the onset of a neurological disorder associated with a lysosomal storage disease, said method comprising: a) measuring the subject's brain tissue volume (e.g., using vMRI) and comparing it with a reference standard to assess whether the brain tissue volume is lower than the reference standard; b) identifying the onset of said neurological disorder if the brain tissue volume identified in step (a) is lower than the reference standard; and c) initiating treatment of the subject by administering to the subject an effective amount of a quinuclidine compound described herein, e.g., a compound according to any of Formulas (I) or (II)-(XII), (Ia)-(XIIa), or (Ib)-(XIIb), or any of Compounds 1 or 1.1-1.75. In an embodiment, the treatment c) can be carried out by any one of Method 1, Method 2, and Method 3 described herein.

[0107] All of the methods described herein (e.g., Method 1, Method 2, Method 3, Method 4, Method 5, as well as any of the embodiments of the above methods) may be useful in subjects who meet one or more or all of the following criteria: a) be 18 years of age or older; b) have a clinical diagnosis of Gaucher disease, e.g., Gaucher disease (e.g., Gaucher disease type 3) Have been diagnosed with or determined to be at risk of developing a disorder, which can be diagnosed by any of the assessments described herein, for example, by having a mutation in a gene described herein; c) a demonstrated deficiency of acid beta-glucosidase activity; d) receiving treatment with an ERT, e.g., imiglucerase (Cerezyme), for at least 3 years with a stable monthly dose for at least 6 months prior to initiating treatment; e) a hemoglobin level of 11.0 g / dL or greater for women and 12.0 g / dL or greater for men; f) Platelet count 100,000 / mm 3 Being more than that; g) spleen volume less than 10 times normal (10MN) or total splenectomy (provided splenectomy occurred more than 3 years prior to randomization); h) liver volume less than 1.5 MN; i) no bone crisis within 3 months or 1 year prior to treatment initiation and no symptomatic bone disease (e.g., bone pain due to osteonecrosis and / or pathological fractures); j) having a history of seizures other than myoclonic seizures; k) Gaucher disease type 3, characterized by oculomotor apraxia (supranuclear gaze paresis) characterized by horizontal saccade abnormalities; and l) Having mild or moderate neurological involvement as measured using the modified Severity Scoring Tool (mSST; Davies, et al., 2011) at the start of treatment.

[0108] In an embodiment of Method 1 described herein, or a specific subembodiment of Method 1 described herein, the subject meets all of criteria (a) to (l). In an embodiment of Method 2 described herein, or a specific subembodiment of Method 2 described herein, the subject meets all of criteria (a) to (l). In an embodiment of Method 3 described herein, or a specific subembodiment of Method 3 described herein, the subject meets all of criteria (a) to (l). In an embodiment of Method 4 described herein, or a specific subembodiment of Method 4 described herein, the subject meets all of criteria (a) to (l). In an embodiment of Method 5 described herein, or a specific subembodiment of Method 5 described herein, the subject meets all of criteria (a) to (l).

[0109] In further embodiments, the subject is an adult or pediatric patient aged 12 years or older. In embodiments, the subject is an adult or pediatric patient aged 12 years or older with Gaucher disease type 3 (e.g., confirmed by meeting criteria b) and / or k) above), whose performance status has been stabilized with an ERT, e.g., imiglucerase (Cerezyme). Performance status can be characterized by the presence of markers of systemic disease, such as those associated with: i) spleen and liver volume (e.g., measured by magnetic resonance imaging (MRI)); ii) platelet count; and iii) hemoglobin level. In some embodiments, the subject with Gaucher disease type 3 has been treated with an ERT (e.g., using imiglucerase (Cerezyme)) for at least 3 years and / or has reached the following GD treatment goals: one or more or all of the above criteria e) to i) are met.

[0110] In a specific embodiment, the present invention provides a single oral dose of 15 mg / day of (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate, in combination with an ERT, e.g., imiglucerase (Cerezil). For use in the methods defined herein (e.g., Method 1, Method 2, Method 3, Method 4, Method 5, and any of the embodiments of said methods described above) in a subject with Gaucher disease type 3 (e.g., confirmed by meeting criteria b) and / or k) above) who has been stabilized with CYME, wherein the subject is an adult or pediatric patient aged 12 years or older.

[0111] As described in Method 1, Method 2, Method 3, and Method 4, respectively, a positive effect on cognitive dysfunction and / or neuronal connectivity and / or brain tissue volume and / or regression of neurological disorders can be understood as treatment of CNS findings. Thus, in a specific embodiment, the present invention provides (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate, at a single oral dose of 15 mg / day for use in treating CNS findings in subjects with Gaucher disease type 3 (e.g., confirmed by meeting criteria b) and / or k) above) who, if in good general condition (as defined above), are stabilized with an ERT, e.g., imiglucerase (Cerezyme), wherein the subjects are adult or pediatric patients aged 12 years or older.

[0112] In each embodiment of the methods described herein (e.g., Method 1, Method 2, Method 3, Method 4, Method 5, and any of the embodiments of the methods described above), the subject being treated is not receiving concomitant therapy with a CYP3A inducer, e.g., a strong CYP3A inducer such as rifampin, or a moderate CYP3A inducer such as phenobarbital or efavirenz. In an embodiment, the subject is not taking a dietary supplement identified as a strong or moderate inducer of CYP3A.

[0113] In one embodiment, the invention provides (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, optionally in the form of an acid addition salt of the malate, at a single oral dose of 15 mg / day for use in treating CNS findings in subjects with Gaucher disease type 3 (e.g., confirmed by meeting criteria b) and / or k) above) who, if in good condition (as defined above), are stabilized with an ERT, e.g., imiglucerase (Cerezyme), where the subject is an adult or pediatric patient 12 years of age or older, and the treated subject is not receiving concomitant therapy with a CYP3A inducer, e.g., a strong CYP3A inducer such as rifampin, or a moderate CYP3A inducer such as phenobarbital or efavirenz. In an embodiment, the subject is not taking dietary supplements identified as strong or moderate inducers of CYP3A.

[0114] All methods described herein (e.g., Method 1, Method 2, Method 3, Method 4, Method 5, and any of the above-mentioned method embodiments) may be deemed inappropriate for certain patient groups, such as those with certain pre-existing conditions, those with current or past treatment with certain medications, and those with a history of treatments such as surgical procedures as evaluated and described herein.The prescribing physician is entitled to determine whether the subject's specific condition and / or current or past medications affect their suitability for treatment according to the methods disclosed herein.

[0115] In embodiments, subjects treated by the methods described herein (e.g., Method 1, Method 2, Method 3, Method 4, Method 5, as well as any of the above method embodiments) do not include subjects who meet one or more or all of the following criteria: 1) had received substrate reduction therapy or chaperone therapy for Gaucher disease within 6 months prior to treatment initiation; 2) had undergone partial or total splenectomy within 3 years prior to the start of treatment; 3) transfusion dependent; 4) Current liver enzymes (alanine aminotransferase [ALT] / aspartate aminotransferase [AST]) or total bilirubin greater than twice the upper limit of normal, unless the patient has previously had esophageal varices or hepatic infarction or has been diagnosed with Gilbert syndrome. 5) have a clinically significant disease other than Gaucher disease, such as cardiovascular (congenital heart disease, coronary artery disease, valvular disease, or left-sided heart failure; clinically significant arrhythmia or conduction defect), hepatic, gastrointestinal, pulmonary, neurological, endocrine, metabolic (e.g., hypokalemia, hypomagnesemia), or psychiatric disease, other medical condition, or serious intercurrent illness that may prevent participation; 6)30mL / min / 1.73m 2 have renal failure as defined by an estimated glomerular filtration rate of less than 7) have a history of cancer except for basal cell carcinoma; 8) have myoclonic seizures; 9) pregnant or breastfeeding; 10) have a cortical cataract exceeding one-quarter of the lens circumference (cortical cataract grade-2) or a posterior subcapsular cataract exceeding 2 mm (posterior subcapsular cataract grade-2) according to the World Health Organization (WHO) grading; 11) the need to use invasive assisted ventilation; 12) the need to use noninvasive assisted ventilation while awake for more than 12 hours each day; 13) currently receiving medications that may cause cataracts (corticosteroids, psoralens used in dermatology with ultraviolet light therapy [PUVA], typical antipsychotics, and glaucoma medications) or that may worsen vision in cataract patients (e.g., alpha-adrenergic glaucoma medications); 14) received a strong or moderate inducer or inhibitor of CYP3A prior to treatment initiation, within 15 days or 5 half-lives from screening, whichever is longer, or consumed grapefruit, grapefruit juice, or grapefruit-containing products within 72 hours of treatment initiation; 15) Patients who are scheduled to be hospitalized during the procedure, including elective surgery; and 16) Have received a major organ transplant (e.g., bone marrow or liver).

[0116] Pharmaceutical Composition The present disclosure also provides pharmaceutical compositions comprising at least one quinuclidine compound described herein and at least one pharmaceutically acceptable excipient, e.g., for use in accordance with the methods disclosed herein. Pharmaceutically acceptable excipients can be found, for example, in the literature, such as those described in Remington's Pharmaceutical Sciences, Mack The excipient may be any excipient known in the art, including those described in A.R. Gennaro Publishing Co. (A.R. Gennaro, ed. 1985). Pharmaceutical compositions of the compounds disclosed herein may be prepared by conventional means known in the art, for example, by mixing at least one compound disclosed herein with a pharmaceutically acceptable excipient.

[0117] Accordingly, in one aspect, the disclosure provides a pharmaceutical dosage form comprising a quinuclidine compound described herein and a pharmaceutically acceptable excipient, wherein the dosage form is formulated to provide a sufficient amount of the compound when administered (e.g., when administered orally) to treat a disease or disorder provided in any of the methods described herein (e.g., Method 1 or later, Method 2 or later, Method 3 or later, Method 4 or later, or Method 5 or later).

[0118] The pharmaceutical composition or dosage form of the present invention comprises a drug and another carrier, e.g., an inert or Active compounds or compositions may include, for example, detectable agents, labels, adjuvants, diluents, binders, stabilizers, buffers, salts, lipophilic solvents, preservatives, adjuvants, etc. Carriers also include pharmaceutical excipients and additives, such as proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and oligosaccharides; derivatized sugars, e.g., alditols, aldonic acids, esterified sugars, etc.; and polysaccharides or sugar polymers), which may be present alone or in combination, including 1 to 99.99% by weight or volume. Exemplary protein excipients include serum albumins, e.g., human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acids / antibody components that can also function in a buffering capacity include alanine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, etc. Carbohydrate excipients are also contemplated within the scope of the present invention, including, but not limited to, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, etc.; disaccharides such as lactose, sucrose, trehalose, cellobiose, etc.; polysaccharides such as raffinose, melezitose, maltodextrin, dextran, starch, etc.; and alditols such as mannitol, xylitol, maltitol, lactitol, xylitol, sorbitol (glucitol), and myo-inositol.

[0119] Carriers that may be used include buffers or pH adjusters; typically, buffers are salts prepared from organic acids or bases. Representative buffers include organic acid salts such as citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid salts; Tris, tromethamine hydrochloride, or phosphate buffers. Additional carriers include polymer excipients / additives such as polyvinylpyrrolidone, Ficoll (polymeric sugar), dextrates (e.g., cyclodextrin, e.g., 2-hydroxypropyl-β-cyclodextrin), polyethylene glycol, flavoring agents, antibacterial agents, sweeteners, antioxidants, antistatic agents, surfactants (e.g., polysorbates, e.g., "TWEEN 20" and "TWEEN 80"), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol), and chelating agents (e.g., EDTA).

[0120] The present disclosure also provides pharmaceutical compositions and kits comprising said compositions, comprising at least one quinuclidine compound described herein and at least one additional pharmaceutically active agent.These pharmaceutical compositions and kits can be adapted to allow the quinuclidine compound and the additional active agent to be administered simultaneously, sequentially, and / or separately.For example, the quinuclidine compound and the additional active agent can be formulated into separate dosage forms, for example, separate tablets, capsules, lyophilized agents, or liquids, or can be formulated into the same dosage form, for example, the same tablet, capsule, lyophilized agent, or liquid.When the quinuclidine compound and the additional active agent are formulated into the same dosage form, the quinuclidine compound and the additional active agent can be substantially present in an additive mixture, for example, in the core of a tablet, or can be substantially present in separate regions of the dosage form, for example, in separate layers of the same tablet. In one embodiment, the pharmaceutical dosage form comprises an additional agent described herein that can treat or prevent cognitive impairment and / or gait abnormalities, e.g., in patients diagnosed with or susceptible to a lysosomal storage disease, e.g., Gaucher Type 3 or Niemann-Pick Type C.

[0121] In a further aspect, the present disclosure provides a pharmaceutical composition comprising: (i) a quinuclidine compound described herein; (ii) an additional active agent; and (iii) a pharmaceutically acceptable excipient. In one embodiment, the additional active agent is a compound described herein, e.g., a compound associated with a lysosomal storage disease, e.g., Gaucher Type 3 or Niemann-Pick Type C. The agent can treat or prevent cognitive impairment and / or gait abnormality in patients who have been diagnosed with or are susceptible to such diseases.In one embodiment, when orally administered to a subject, the additional active agent can treat or prevent gait disorder (e.g., ataxia) or dementia in patients who have been diagnosed with or are susceptible to such diseases, for example, lysosomal storage disease, as described herein, for example, Gaucher type 3 or Niemann-Pick type C.

[0122] The quinuclidine compounds and pharmaceutical compositions disclosed herein may be used in animals or humans.Thus, the compounds disclosed herein may be formulated as pharmaceutical compositions for oral, buccal, parenteral (e.g., intravenous, intramuscular, or subcutaneous), topical, rectal, or intranasal administration, or in a form suitable for administration by inhalation or insufflation.In certain embodiments, the quinuclidine compounds or pharmaceutical compositions are formulated for systemic administration, for example, via a route other than parenteral.In one embodiment, the quinuclidine compounds or pharmaceutical compositions are formulated for oral administration, for example, in solid form.Such modes of administration and methods for preparing suitable pharmaceutical compositions are described, for example, in Gibaldi's Drug Delivery Systems in Pharmaceutical Care (1st ed., American Society of Health-System Pharmacists 2007).

[0123] Pharmaceutical compositions may be formulated to delay, extend, or control the release of the active ingredient therein, for example, using various proportions of hydroxypropylmethylcellulose, other polymer matrices, liposomes, and / or microspheres to provide the desired release profile. The pharmaceutical compositions may also optionally contain opacifying agents, and may optionally be compositions that release the active ingredient only or preferentially in a specific portion of the gastrointestinal tract in a delayed manner, for example, by using an enteric coating. Examples of implant compositions include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, where appropriate, with one or more pharmaceutically acceptable carriers, excipients, or diluents known in the art (see, e.g., Remington's). The compounds disclosed herein may also be formulated for sustained delivery according to methods known to those skilled in the art. Examples of such formulations can be found in U.S. Patent Nos. 3,119,742; 3,492,397; 3,538,214; 4,060,598; and 4,173,626.

[0124] In solid dosage forms for oral administration (e.g., capsules, tablets, pills, dragees, powders, granules, and the like), the active ingredient may be combined with one or more pharmaceutically acceptable carriers, excipients, or diluents, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, microcrystalline cellulose, calcium phosphate, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginate, gelatin, pregelatinized maize starch, polyvinylpyrrolidone, hydroxypropyl methylcellulose, sucrose, and / or acacia; (3) humectants, such as glutaraldehyde, glycerin, PEG-10 ... Glycerol; (4) disintegrants, such as agar-agar, calcium carbonate, sodium starch glycolate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retardants, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as sodium lauryl sulfate, acetyl alcohol, and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, silica, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets, and pills, the pharmaceutical compositions may also contain buffering agents. Similar types of solid compositions may also be used. The composition may also be prepared in soft and hard-filled gelatin capsules using fillers and excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

[0125] Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface-active agents, and / or dispersants. Molded tablets may be made by molding a mixture of powdered active ingredient moistened with an inert liquid diluent in a suitable machine. Tablets and other solid dosage forms, such as sugar-coated tablets, capsules, pills, and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the art.

[0126] In some embodiments, pharmaceutical compositions are orally administered in liquid form.Liquid dosage forms for oral administration of active ingredients include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.Liquid preparations for oral administration can also be in the form of a dry product, which can be reconstituted with water or other suitable vehicles before use.In addition to active ingredients, liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (such as cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, liquid pharmaceutical compositions may contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweeteners, flavorings, coloring agents, perfumes, and preservatives.In addition to the active ingredient, suspensions may contain suspending agents, such as, but not limited to, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.Suitable liquid preparations may be prepared by conventional means using pharmaceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, methylcellulose, or hydrogenated edible fats); emulsifiers (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, or ethyl alcohol); and / or preservatives (e.g., methyl or propyl p-hydroxybenzoate or sorbic acid). The active ingredient may also be administered as a bolus, electuary or paste.

[0127] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0128] In embodiments, the pharmaceutical composition is administered by parenteral means, e.g., topical application, transdermal application, injection, etc. In related embodiments, the pharmaceutical composition is administered parenterally by injection, infusion, or implantation (e.g., intravenous, intramuscular, intraarterial, subcutaneous, etc.).

[0129] The compounds disclosed herein may be formulated for parenteral administration by injection, including using conventional catheterization techniques or infusion. The injectable formulations may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may be prepared in an oil-based formulation. The active ingredient may take such forms as suspensions, solutions, or emulsions in aqueous or non-aqueous vehicles and may contain formulatory agents recognized by those skilled in the art, such as suspending, stabilizing, and / or dispersing agents. Alternatively, the active ingredient may be in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0130] The pharmaceutical composition may be administered directly to the central nervous system. Thus, in certain embodiments, the composition is administered directly to the central nervous system, thereby bypassing the blood-brain barrier. In some embodiments, the composition may be administered via direct spinal injection. In embodiments, the composition is administered by intrathecal injection. In some embodiments, the composition is administered via intraventricular injection. In embodiments, the composition is administered to a lateral ventricle. In embodiments, the composition is administered to both lateral ventricles. In additional embodiments, the composition is administered via intrahippocampal injection. The composition may be administered in a single injection or multiple injections. In other embodiments, the composition is administered to more than one location (e.g., two locations in the central nervous system).

[0131] Pharmaceutical compositions may be in sterile injectable form. Pharmaceutical compositions may be rendered sterile, for example, by filtration through a bacterial-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in water or some other sterile injectable medium immediately before use. To prepare such compositions, the active ingredient is dissolved or suspended in a parenterally acceptable liquid vehicle. Exemplary vehicles and solvents include, but are not limited to, water, water adjusted to a suitable pH by adding an appropriate amount of hydrochloric acid, sodium hydroxide, or a suitable buffer, 1,3-butanediol, Ringer's solution, and isotonic sodium chloride solution. Pharmaceutical compositions may also contain one or more preservatives, such as methyl, ethyl, or n-propyl p-hydroxybenzoates. To improve solubility, a solution-enhancing agent or solubilizer may be added, or the solvent may contain 10-60% w / w propylene glycol or a similar substance.

[0132] Pharmaceutical compositions may comprise one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately prior to use. Such pharmaceutical compositions may also contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, suspending agents, thickening agents, preservatives, etc.

[0133] Examples of suitable aqueous and non-aqueous carriers that may be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. In some embodiments, to prolong the effect of an active ingredient, it is desirable to delay absorption of the compound from subcutaneous or intramuscular injection. This may be achieved by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the active ingredient then depends upon its rate of dissolution, which may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered active ingredient can be achieved by dissolving or suspending the compound in an oil vehicle. Furthermore, prolonged absorption of injectable pharmaceutical forms can be achieved by including agents that delay absorption, for example, aluminum monostearate and gelatin.

[0134] Controlled release parenteral compositions include aqueous suspensions, microspheres, microcapsules, magnetic The active ingredient may be in the form of an aqueous microsphere, oil solution, oil suspension, or emulsion, or the active ingredient may be incorporated into a biocompatible carrier, liposome, nanoparticle, implant, or infusion device. Materials for use in preparing microspheres and / or microcapsules include, but are not limited to, biodegradable / bioerodible polymers such as polyglactin, poly-(isobutyl cyanoacrylate), poly(2-hydroxyethyl-L-glutamine), and poly(lactic acid). Biocompatible carriers that may be used when formulating controlled-release parenteral formulations include carbohydrates, such as dextran, proteins, such as albumin, lipoproteins, or antibodies. Materials for use in implants may be non-biodegradable, such as polydimethylsiloxane, or biodegradable, such as poly(caprolactone), poly(lactic acid), poly(glycolic acid), or poly(orthoesters).

[0135] For topical administration, the compounds disclosed herein may be formulated into ointments or creams. The compounds disclosed herein may also be formulated in rectal compositions such as suppositories or retention enemas, for example, containing conventional suppository bases such as cocoa butter or other glycerides.

[0136] For intranasal administration or inhalation administration, the compounds disclosed herein can be conveniently delivered in the form of a solution or suspension from a pump spray container that is pushed or pumped by the patient, or as an aerosol spray presented from a pressurized container or nebulizer using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas.In the case of pressurized aerosol, dosage units can be determined by providing a valve to deliver a metered amount.The pressurized container or nebulizer can contain a solution or suspension of the compounds disclosed herein.Inhalers or insufflators for use in capsules and cartridges (e.g., made from gelatin) can be formulated to contain a powder mix of the compounds disclosed herein and a suitable powder base, for example, lactose or starch.

[0137] Generally, the medicament and composition described herein is administered in an effective amount or amount that is sufficient to treat or prevent cognitive dysfunction and / or gait abnormality in the subject who needs it.Typically, dosage can be adjusted within this range based on, for example, age, physical condition, weight, sex, diet, administration time and other clinical factors.The determination of effective amount is well within the ability of those skilled in the art.

[0138] Having generally described the present invention, the following non-limiting examples and accompanying figures are provided to further illustrate the invention. [Brief explanation of the drawings]

[0139] [Figure 1] 1 is a graph showing the change in mean whole brain volume (WBV) after 52 weeks of treatment with the compound of formula (I) for Group A patients, and the change in WBV for patient 5, as determined from vMRI measurements and TBM analysis according to Example 5 herein. [Example]

[0140] General Procedure for Chemical Synthesis General Procedure A: Carbamate Formation with Triphosgene To a suspension of amine hydrochloride (1 equiv.) and triethylamine (3-4 equiv.) in THF (approximately 0.2 M concentration) at room temperature, triphosgene (0.35 equiv.) was added. The reaction mixture was stirred for 10 min, and a small amount of ether (1-2 mL) was added. The triethylammonium salt was filtered off to give a clear solution of the isocyanate in THF / ether.

[0141] To a solution of the alcohol (1.5 equiv.) in THF (concentration approximately 0.2 M) at room temperature was added NaH [60%, oil] (1.5 equiv.). The reaction mixture was stirred for 15 min, and the above solution (isocyanate in THF / ether) was added dropwise. For standard workup, the reaction was quenched with brine. The solution was extracted with EtOAc, and the organic layer was dried over Na2SO4, filtered, and concentrated. The crude material was purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding carbamate.

[0142] General Procedure B: Alkylation with Organocerium A suspension of CeCl3 (4 equiv.) in THF (approximately 0.2 M) was stirred at room temperature for 1 h. The suspension was cooled to -78 °C, and 1.6 M MeLi / ether (4 equiv.) was added dropwise. The organocerium complex was allowed to form for 1 h, and a solution of the nitrile (1 equiv.) in 2.0 M THF was added dropwise. The reaction mixture was warmed to room temperature and stirred for 18 h. The solution was cooled to 0 °C and quenched with water (approximately 1 mL), followed by the addition of 50% aqueous ammonium hydroxide (approximately 3 mL) until a precipitate formed and sank to the bottom of the flask. The mixture was filtered through a pad of Celite and concentrated. The crude material was treated with 4.0 M HCl / dioxane solution. The intermediate arylpropan-2-amine hydrochloride was triturated with ether and used directly in the next step. Alternatively, the crude free-based amine was purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding arylpropylamine.

[0143] General Procedure C: Suzuki Coupling To a solution of aryl halide (1 equiv.) in a 4:1 mixture of DME / water (approximately 0.2 M concentration) was added boronic acid (2 equiv.), palladium catalyst (0.1–0.25 equiv.), and sodium carbonate (2 equiv.). The reaction mixture was microwaved at 150 °C for 25 min. After filtration through a Celite plug and concentration, the crude product was purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding coupling adduct.

[0144] Alternative method: To a solution of aryl halide (1 equiv.) in a toluene / water [20:1] mixture (approximately 0.2 M concentration), boronic acid (1.3–2.5 equiv.), palladium catalyst (0.05–0.15 equiv.), tricyclohexylphosphine (0.15–0.45 equiv.), and potassium phosphate (5 equiv.) were added. The reaction mixture was microwaved at 150 °C for 25 min. After filtration through a Celite plug and concentration, the crude product was purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding coupling adduct.

[0145] General Procedure D: Cyclopropanation To a mixture of arylnitrile (1 equiv.) and Ti(Oi-Pr) (1.7 equiv.) stirred at -70 °C, EtMgBr (3.0 M in ether) (1.1 equiv.) was added dropwise. The reaction mixture was warmed to 25 °C and stirred for 1 h. BF·EtO (3 equiv.) was added dropwise to the above mixture at 25 °C. After the addition, the mixture was stirred for an additional 2 h and then quenched with aqueous HCl (2 M). The resulting solution was then basified by adding aqueous NaOH (2 M). The organic material was extracted with ethyl ether. The organic layers were combined, dried over NaSO, filtered, and concentrated. The crude material was purified by silica gel column chromatography (eluting with petroleum ether / EtOAc: 10 / 1 to 1 / 1) to give the corresponding 1-aryl-cyclopropanamine.

[0146] General Procedure E: Biaryl Coupling Using Suzuki Conditions 5:1 (v / v) dioxane / water (approximately 0.15 M) or 5:1 (v / v) N,N To a stirred solution of aryl halide component (1 equivalent) in dimethylformamide (approximately 0.15 M) was added aryl boronate or aryl boronic acid component (1-1.5 equivalents), sodium carbonate (2-3 equivalents), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.05 equivalents). The mixture was heated (90 °C) overnight and then filtered through a Celite plug. The Celite was rinsed with ethyl acetate, and the combined filtrate was washed with brine, dried (Na2SO4), and concentrated. The residue was purified by silica flash chromatography.

[0147] General Procedure F: Carbamate formation using isocyanates generated via the mixed anhydride / Curtius rearrangement route To a stirred solution of the carboxylic acid component (1 equivalent) in tetrahydrofuran (approximately 0.1 M) was added triethylamine (2 equivalents). The reaction was cooled (0 °C) and treated with isobutyl chloroformate (1.5 equivalents). After 1 h at 0 °C, a solution of sodium azide (2 equivalents) in water (approximately 1 M) was added, and the reaction was allowed to warm to room temperature. After stirring overnight, the reaction was diluted with water and extracted with ethyl acetate. The combined extracts were washed with aqueous sodium bicarbonate and brine, dried (Na2SO4), and concentrated. The crude acyl azide was further dried via coevaporation with toluene and then taken up in toluene (approximately 0.1 M). The stirred solution was refluxed for 2–2.5 h, cooled, and treated with the alcohol component (1.25–2 equivalents). The reaction was heated to reflux overnight and then concentrated. The residue was taken up in either ethyl acetate or chloroform, washed with aqueous sodium carbonate (Na2SO4), and concentrated. The crude product was purified by silica flash chromatography using a chloroform / methanol (less polar carbamates) or chloroform / methanol / ammonia (more polar carbamates) solvent gradient. Example 1

[0148] Synthesis of quinuclidine compounds 1-Azabicyclo[2.2.2]oct-3-yl[2-(4'-fluorobiphenyl-3-yl)propan-2-yl]carbamate (Compound 1) Using general procedure C, 1-azabicyclo[2.2.2]oct-3-yl[2-(3-bromophenyl)propan-2-yl]carbamate (600 mg, 1.63 mmol), 4-fluorophenylboronic acid (457 mg, 3.27 mmol), and palladium(II) acetate gave the title compound as a white solid (373 mg; 60%). 1 H NMR (400 MHz, CDCl3) δ 7.56 (s, 1H), 7.52 (dd, J = 5.4, 8.4 Hz, 2H), 7.42-7.38 (m, 3H), 7.12 (m, 2H), 5.18 (5, 1H), 4.62 (s, 1H), 2.66 (m, 6H), 1.72 (s, 6H), 2.01-0.83 (m, 5H) ppm. 13 C NMR (100 MHz, CDCl3) δ 125.0, 124.0, 123.8, 116.0, 116.0, 71.3, 55.9, 55.5, 47.6, 46.7, 29.6, 25.6, 24.8, 19.8 ppm. Purity: 98.0% UPLCMS (210 nm); retention time 0.95 min; (M+1) 382.9. Anal. Calcd. for C 23 H 27 FN2O2·0.37(CHCl3): C, 65.86; H, 6.47; N, 6.57. Found: C, 65.85; H, 6.69; N, 6.49.

[0149] (S)-Quinuclidin-3-yl 2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-ylcarbamate (Compound 2) To a stirred solution of 4-fluorothiobenzamide (8.94 g, 57.6 mmol) in ethanol (70 mL) was added ethyl 4-chloroacetate (7.8 mL, 58 mmol). The reaction was heated to reflux for 4 h, treated with an aliquot of ethyl 4-chloroacetoacetate (1.0 mL, 7.4 mmol), and refluxed for an additional 3.5 h. The reaction was then concentrated, and the residue was partitioned between ethyl acetate (200 mL) and aqueous NaHCO (200 mL). The organic layer was combined with a back-extraction of the aqueous layer (ethyl acetate, 1 × 75 mL), dried (Na SO ), and concentrated. The resulting amber oil was purified by flash chromatography using a hexane / ethyl acetate gradient to give ethyl 2-(2-(4-fluorophenyl)thiazol-4-yl)acetate as a low-melting, nearly colorless solid. (13.58g, 89%)

[0150] To a stirred solution of ethyl 2-(2-(4-fluorophenyl)thiazol-4-yl)acetate (6.28 g, 23.7 mmol) in DMF (50 mL) was added sodium hydride [60% dispersion in mineral oil] (2.84 g, 71.0 mmol). The foamy mixture was stirred for 15 minutes, then cooled in an ice bath and iodomethane (4.4 mL, 71 mmol) was added. The reaction was stirred overnight, allowing the cooling bath to slowly warm to room temperature. The mixture was then concentrated and the residue was partitioned between ethyl acetate (80 mL) and water (200 mL). The organic layer was washed with a second portion of water (1 × 200 mL), dried (NaSO), and concentrated. The resulting amber oil was purified by flash chromatography using a hexane / ethyl acetate gradient to give ethyl 2-(2-(4-fluorophenyl)thiazol-4-yl)-2-methylpropanoate as a colorless oil (4.57 g, 66%).

[0151] To a stirred solution of ethyl 2-(2-(4-fluorophenyl)thiazol-4-yl)-2-methylpropanoate (4.56 g, 15.5 mmol) in 1:1:1 THF / ethanol / water (45 mL) was added lithium hydroxide monohydrate (2.93 g, 69.8 mmol). The reaction was stirred overnight, concentrated, and redissolved in water (175 mL). The solution was washed with ether (1 × 100 mL), acidified by adding 1.0 N HCl (80 mL), and extracted with ethyl acetate (2 × 70 mL). The combined extracts were dried (NaSO) and concentrated to give 2-(2-(4-fluorophenyl)thiazol-4-yl)-2-methylpropanoic acid as a white solid (4.04 g, 98%). This material was used in the next step without purification.

[0152] To a stirred, cooled (0 °C) solution of 2-(2-(4-fluorophenyl)thiazol-4-yl)-2-methylpropanoic acid (4.02 g, 15.2 mmol) in THF (100 mL) was added trimethylamine (4.2 mL, 30 mmol), followed by isobutyl chloroformate (3.0 mL, 23 mmol). The reaction was stirred cold for an additional 1 h before a solution of sodium azide (1.98 g, 30.5 mmol) in water (20 mL) was added. The reaction was stirred overnight, allowing the cooling bath to slowly warm to room temperature. The mixture was then diluted with water (100 mL) and extracted with ethyl acetate (2 × 60 mL). The combined extracts were washed with aqueous NaHCO (1 × 150 mL) and brine (1 × 100 mL), dried (NaSO), and concentrated. After coevaporation with toluene (2 × 50 mL), the resulting white solid was taken up in toluene (100 mL) and refluxed for 4 hours. (S)-3-quinuclidinol (3.87 g, 30.4 mmol) was then added, and refluxing was continued overnight. The reaction was concentrated, and the residue was partitioned between ethyl acetate (100 mL) and aqueous NaHCO (150 mL). The organic layer was washed with water (1 × 150 mL), dried (Na SO ), and concentrated. The resulting off-white solid was purified by flash chromatography using a chloroform / methanol / ammonia gradient to give the title compound as a white solid (4.34 g, 73%). 1H NMR (400 MHz, CDCl3) δ 7.96-7.88 (m, 2H), 7.16-7.04 (m, 3H), 5.55 (br s, 1H), 4.69-4.62 (m, 1H), 3.24-3.11 (m, 1H), 3.00-2.50 (m, 5H), 2.01-1.26 (m, 11H) ppm. 13 C NMR (400 MHz, CDCl3) δ 166.4, 165.1, 163.8 (d, J=250.3 Hz), 162.9, 155.0, 130.1 (d, J=3.3 Hz), 128.4 (d, J= 8.5 Hz), 115.9 (d, J= 22.3 (Hz), 112.5, 71.2, 55.7, 54.2, 47.5, 46.5, 28.0, 25.5, 24.7, 19.6 ppm. Purity: 100 % UPLCMS (210 nm & 254 nm); retention time 0.83 min; (M+1) 390.

[0153] (S)-Quinuclidin-3-yl(2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 3) Using general procedure E, the reaction used ethyl 2-(4-bromophenyl)-2-methylpropanoate and 4-(2-methoxyethoxy)phenylboronic acid (input), and ethyl 2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)-2- Methyl propanoate was prepared as an off-white solid. To a stirred solution of this compound (3.01 g, 8.78 mmol) in 1:1:1 (v / v / v) tetrahydrofuran / ethanol / water (45 mL) was added lithium hydroxide monohydrate (1.47 g, 61.4 mmol). The mixture was heated to reflux overnight and then concentrated. The residue was dissolved in water, treated with 1N hydrochloric acid (65 mL), and extracted with ethyl acetate. The combined organic layers were washed with brine, dried (NaSO), and concentrated to give 2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)-2-methylpropanoic acid as a white solid (2.75 g, 100%). This intermediate and (S)-quinuclidin-3-ol were reacted according to General Procedure F to produce the title compound as a colorless glassy solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.62-7.29 (m, 7H), 7.01 (d, J = 8.9 Hz, 2H), 4.47-4.37 (m, 1H), 4.17-4.08 (m, 2H), 3.72-3.62 (m, 2H), 3.32 (s, 3H), 3.09-2.25 (m, 6H), 2.05-1.18 (m, 11H) ppm. 13 C NMR (100 MHz, DMSO-d6) δ 157.9, 154.5, 146.7, 137.4, 132.5, 127.5, 125.7, 125.2, 114.8, 70.4, 70.0, 66.9, 58.2, 55.4, 54.2, 46.9, 45.9, 29.4, 25.3, 24.2, 19.2 ppm. Purity: 100%, 100% (210 & 254 nm) UPLCMS; retention time: 0.87 min; (M+H + ) 439.5.

[0154] 1-Azabicyclo[2.2.2]oct-3-yl[2-(biphenyl-3-yl)propan-2-yl]carbamate (Compound 4) Using general procedure C, 1-azabicyclo[2.2.2]oct-3-yl[2-(3-bromophenyl)propan-2-yl]carbamate (600 mg, 1.63 mmol), phenylboronic acid (398 mg, 3.27 mmol), and palladium(II) acetate gave the title compound as a white solid (379 mg, 64%). 1 H NMR (400 MHz, CDCl3) δ 7.61 (s, 1H), 7.56 (d, J= 7.4 Hz, 2H), 7.50-7.38 (m, 4H), 7.34 (m, 2H), 5.16 (s, 1H), 4.63 (s, 1H), 3.39-2.09 (m, 6H), 1.72 (s, 6H), 2.02-0.73 (m, 5H) ppm. 13 C NMR (100 MHz, CDCl3) δ 154.8, 147.8, 141.6, 129.0, 129.0, 128.6, 127.5, 125.8, 125.0, 124.0, 71.6, 71.3, 55.9, 55.5, 47.6, 46.8, 31.5, 30.2, 30.0, 29.5, 25.6, 24.8, 19.8 ppm. Purity: 99% UPLCMS (210 nm); retention time 0.84 min; (M+1) 365.0. Anal. Calcd. for C 23 H 28 N2O2·0.29(CHCl3): C, 70.02; H, 7.14; N, 7.01. Found: C, 70.02; H, 7.37; N, 6.84.

[0155] (S)-Quinuclidin-3-yl 2-(biphenyl-4-yl)propan-2-ylcarbamate (Compound 5) Using general procedure B, bromobenzonitrile (2.00 g, 11.0 mmol) was converted to the corresponding 2-(4-bromophenyl)propan-2-amine (1.20 g, 51%) as a brown oil.

[0156] Using general procedure A, 2-(4-bromophenyl)propan-2-amine (1.0 g, 4.7 mmol) and (S)-quinuclidin-3-ol gave (S)-quinuclidin-3-yl 2-(4-bromophenyl)propan-2-ylcarbamate (1.0 g, 58%) as a brown oil.

[0157] Using general procedure C, the above bromide (200 mg, 0.540 mmol), phenylboronic acid (133 mg, 1.10 mmol), and [PdCl 2 (pddf)]CH 2 Cl 2 gave the title compound as a white solid (70 mg, 35%). 1 H NMR (500 MHz, CDCl3) δ 7.60-7.53 (m, 4H), 7.47 (d, J = 8.5 Hz, 2H), 7.42 (t, J = 7.5 Hz, 2H), 7.33 (t, J = 7.5 Hz, 1H), 5.26 (br s, 1H), 4.64 (m, 1H), 3.33-3.15 (m, 1H), 3.10-2.45 (m, 5H), 2.40-1.80 (m, 2H), 1.78-1.58 (m, 7H), 1.55-1.33 (m, 2H) ppm. 13 C NMR (125 MHz, CDCl3) δ154.5, 146.1, 140.8, 139.5, 128.7, 127.2, 127.1, 127.1, 125.2, 70.9, 55.5, 55.1, 47.4, 46.4, 31.1, 29.5, 25.3, 24.5, 19.5 ppm. Purity: 100 % LCMS (214 nm & 254 nm); retention time 1.56 min; (M+1) 365.

[0158] Quinuclidin-3-yl 1-(biphenyl-4-yl)cyclopropylcarbamate (Compound 6) Using general procedure D, bromobenzonitrile (3.00 g, 16.5 mmol) was converted to the corresponding 1-(4-bromophenyl)cyclopropanamine (1.80 g, 51%) as a yellow solid.

[0159] Using general procedure A, 1-(4-bromophenyl)cyclopropanamine (1.0 g, 4.7 mmol) and quinuclidin-3-ol gave quinuclidin-3-yl 1-(4-bromophenyl)cyclopropyl-carbamate (1.3 g, 75%) as a white semi-solid.

[0160] Using general procedure C, the above carbamate (400 mg, 1.12 mmol), phenylboronic acid (267 mg, 2.22 mmol), and [PdCl 2 (pddf)]CH 2 Cl 2 gave the title compound as a viscous oil (100 mg, 25%). 1 H NMR (500 MHz, CDCl3) δ 7.47 (d, J= 7.5 Hz, 2H), 7.43 (d, J= 8.0 Hz, 2H), 7.33 (t, J= 7.5 Hz, 2H), 7.26-7.15 (m, 3H), 5.93 (br s, 0.6H), 5.89 (br s, 0.4H), 4.67 (m, 1H), 3.20-3.06 (m, 1H), 2.88-2.42 (m, 5H), 1.98-1.08 (m, 9H) ppm. 13 C NMR (125 MHz, CDCl3) δ 155.0, 141.0, 139.7, 138.2, 127.7, 126.1, 126.0, 124.8, 124.1, 70.0, 54.5, 46.3, 45.4, 34.1, 24.3, 23.2, 18.3, 17.0 ppm. Purity: 100 % LCMC (214 nm & 254 nm); retention time 1.52 min; (M+1) 363.

[0161] (S)-Quinuclidin-3-yl 1-(4'-fluorobiphenyl-4-yl)cyclopropylcarbamate (Compound 7) Using general procedure C, (S)-quinuclidin-3-yl 1-(4-bromophenyl)cyclopropylcarbamate, 4-F-phenylboronic acid, and [PdCl2(pddf)]CH2Cl2 gave the title compound as a white solid (45%). 1 H NMR (500 MHz, DMSO-d6) δ 8.06-7.83 (d, 1H), 7.69-7.66 (m, 2H), 7.59-7.55 (m, 2H), 7.29-7.22 (m, 4H), 4.56-4.54 (m, 1H), 3.13-2.32 (m, 6H), 1.91-1.19 (m, 9H) ppm. 13 C NMR (125 MHz, DMSO-d6) δ 163.2, 161.2, 156.4, 143.7, 136.9, 128.9, 128.8, 126.8, 125.6, 116.2, 116.0, 70.7, 55.8, 47.4, 46.4, 34.8, 25.7, 24.6, 19.6, 18.7, 18.6 ppm. Purity: > 97 % LCMS (214 nm & 254 nm); retention time 1.96 min; (M+1) 381.2.

[0162] (S)-1-Azabicyclo[2.2.2]oct-3-yl[1-(2',4'-difluorobiphenyl-4-yl)cyclopropyl]carbamate (Compound 8) Using general procedure C, (S)-quinuclidin-3-yl 1-(4-bromophenyl)cyclopropylcarbamate (0.446 g, 1.22 mmol), 2,4-difluorophenylboronic acid (0.386 g, 2.44 mmol) and Pd(OAc) (0.015 g, 0.067 mmol) gave the title compound as a tan solid (0.111 g, 23%). 1H NMR (CDCl3) δ 7.43 (dd, J = 8.4, 1.6 Hz, 2H), 7.40-7.33 (m, 1H), 7.31 (d, J = 7.7 Hz, 2H), 6.99-6.81 (m, 2H), 5.54 (d, J = 48.0 Hz, 1H), 4.82-4.65 (m, 1H), 3.30-3.07 (m, 1H), 2.98-2.44 (m, 5H), 1.97 (d, J = 32.7 Hz, 1H), 1.83 (d, J = 10.3 Hz, 1H), 1.64 (s, 1H), 1.52 (s, 1H), 1.39 (s, 1H), 1.31 (d, J = 6.8 Hz, 4H) ppm. 13 C NMR major rotomer (CDCl3) δ 162.2 (dd, J = 12.8, 249.1 Hz), 159.8 (dd, J = 11.8, 251.0 Hz), 156.9, 156.0, 142.6, 133.1, 131.3 (m), 128.9, 125.6, 124.9, 111.5 (dd, J = 3.9, 21.2 Hz) 104.4 (dd, J = 25.2, 29.4 Hz), 72.1, 71.6, 55.7, 47.4, 46.5, 35.7, 35.3, 25.5, 24.6, 24.4, 19.5, 18.1 ppm. Purity: LCMS > 99.3 % (214 nm & 254 nm); retention time 0.90 min; (M+1) 399.0.

[0163] 1-Azabicyclo[2.2.2]oct-3-yl[1-(4'-methoxybiphenyl-4-yl)cyclopropyl]carbamate (Compound 9) Using general procedure C, quinuclidin-3-yl 1-(4-bromophenyl)cyclopropylcarbamate (0.485 g, 1.33 mmol), 4-methoxyphenylboronic acid (0.404 g, 2.66 mmol), and Pd(OAc) (0.016 g, 0.071 mmol) gave the title compound as a gray solid (0.337 mg, 65%). 1 H NMR (CDCl3) δ 7.48 (dd, J = 8.6, 5.5 Hz, 4H), 7.29 (d, J = 7.6 Hz, 2H), 6.96 (d, J = 8.8 Hz, 2H), 5.58 (d, J = 48.7 Hz, 1H), 4.83-4.63 (m, 1H), 3.84 (s, 3H), 3.20 (dd, J = 24.0, 15.5 Hz, 1H), 2.97-2.42 (m, 5H), 1.97 (d, J = 30.9 Hz, 1H), 1.81 (s, 1H), 1.75-1.33 (m, 3H), 1.28 (d, J = 6.8 Hz, 4H) ppm. 13 C NMR major rotomer (CDCl3) δ 159.1, 156.0, 141.4, 139.0, 133.4, 128.0, 126.7, 125.9, 114.2, 71.5, 55.7, 55.3, 47.4, 46.5, 35.3, 25.5, 24.6, 19.6, 17.8 ppm. Purity: LCMS >97.1 % (214 nm & 254 nm); retention time 0.88 min; (M+1) 393.4.

[0164] Quinuclidin-3-yl 2-(5-(4-fluorophenyl)thiophen-3-yl)propan-2-ylcarbamate (Compound 10) To a stirred, cooled (0 °C) solution of ethyl 5-bromothiophene-3-carboxylate (13.30 g, 56.57 mmol) in THF (100 mL) was added dropwise over 20 min a solution of methylmagnesium bromide in diethyl ether [3.0 M] (55.0 mL, 165 mmol). After 2 h, the reaction solution was concentrated. The residue was taken up in aqueous NH4Cl (200 mL) and extracted with ethyl acetate (2 × 100 mL). The combined extracts were dried (Na2SO4) and concentrated. The resulting amber oil was purified by flash chromatography using a hexane / ethyl acetate gradient to give 2-(5-bromothiophen-3-yl)propan-2-ol as a light amber oil (8.05 g, 64%).

[0165] To a stirred solution of 2-(5-bromothiophen-3-yl)propan-2-ol (8.03 g, 36.3 mmol) in methylene chloride (80 mL) was added sodium azide (7.08 g, 109 mmol), followed by trifluoroacetic acid (8.0 mL; dropwise addition over 5–6 min). The thick suspension was stirred for 1.5 h, then diluted with water (350 mL) and extracted with ethyl acetate (1 × 200 mL). The organic layer was washed with aqueous NaHCO (1 × 250 mL), dried (NaSO), and concentrated to give the crude azide product. To a stirred solution of this material in THF (160 mL) was added water (11 mL), followed by triphenylphosphine (23.8 g, 90.7 mmol). The reaction was stirred for 2 days and then concentrated. The resulting residue was dissolved in ethyl acetate (250 mL) and extracted with 1 N aqueous HCl (4 × 75 mL). The combined extracts were basified with concentrated NHOH and extracted with ethyl acetate (2 × 100 mL). These extracts were dried (NaSO) and concentrated. The resulting amber oil was purified by flash chromatography using a methylene chloride / methanol / ammonia gradient to yield a mixture of 2-(5-bromothiophen-3-yl)propan-2-amine and triphenylphosphine oxide (approximately 70 / 30 ratio) as a viscous amber oil (1.32 g, 17%).

[0166] To a stirred solution of 3-quinuclidinol (3.00 g, 23.6 mmol) in THF (100 mL) was added 4-nitrophenyl chloroformate (5.94 g, 29.5 mmol). After stirring for 4 h, the precipitate was filtered off, rinsed with THF, and air-dried on a frit under house vacuum. The filtrate was dissolved in ethyl acetate (150 mL) and washed with aqueous NaHCO (1 × 150 mL) and water (2 × 150 mL). The organic layer was dried (NaSO) and concentrated to give the crude 4-nitrophenyl quinuclidin-3-yl carbonate product, which was used in the next step without purification.

[0167] 2-(5-bromothiophen-3-yl)propan-2-amine in THF (10 mL) To a stirred solution of quinuclidin-3-yl carbonate (0.366 g, 1.66 mmol) was added 4-nitrophenyl quinuclidin-3-yl carbonate (0.571 g, 1.95 mmol) and a small amount of granular 4-(dimethylamino)pyridine. The mixture was refluxed overnight, concentrated, and partitioned between ethyl acetate (50 mL) and aqueous NaHCO (50 mL). The organic layer was washed again with aqueous NaHCO (1 × 50 mL), dried (NaSO), and concentrated. The resulting dull yellow gum was purified by flash chromatography using a chloroform / methanol / ammonia gradient to give quinuclidin-3-yl (1-(5-bromothiophen-3-yl)cyclopropyl)carbamate as an off-white solid (0.305 g, 49%).

[0168] Using general procedure C, quinuclidin-3-yl(1-(5-bromothiophen-3-yl)cyclopropyl)carbamate (0.227 g, 0.742 mmol), 4-fluorophenylboronic acid (0.208 g, 1.49 mmol), tricyclohexylphosphine (0.021 g, 0.075 mmol), potassium phosphate (0.866, 4.08 mmol), and palladium acetate (8.0 mg, 36 μmol) gave the title compound as a gray solid (0.142 g, 49%). 1 H NMR (400 MHz, CDCl3) δ 7.60-7.45 (m, 2H), 7.24-7.19 (m, 1H), 7.10-6.97 (m, 3H), 5.23 (br s, 1H), 4.72-4.61 (m, 1H), 3.30-3.04 (m, 1H), 3.03-2.25 (m, 5H), 2.09-1.02 (m, 11H) ppm. 13 C NMR (400 MHz, CDCl3) δ 162.3 (d, J = 247.1 Hz), 154.5, 149.8, 143.6, 130.7, 127.4 (d, J = Purity: 95.8 % UPLCMS (210 nm) & 254 nm); retention time 0.90 min; (M+1) 389.

[0169] (S)-Quinuclidin-3-yl 2-(3-(4-fluorophenyl)isothiazol-5-yl)propan-2-ylcarbamate (Compound 11) To a stirred solution of 2-(3-(4-fluorophenyl)isothiazol-5-yl)propan-2-amine (1.21 g, 5.12 mmol) in toluene was added a solution of phosgene in toluene [approximately 1.9 M] (10.8 mL, 20.5 mmol). The reaction was heated to reflux for 2 hours and then concentrated. The residue was co-evaporated with toluene (2 × 15 mL) to give the crude isocyanate intermediate as a golden oil. This material was taken up in toluene (10 mL) and treated with (S)-3-quinuclidinol (0.749 g, 5.89 mmol). The reaction was heated to reflux overnight and concentrated. The residue was purified by flash chromatography using a chloroform / methanol / ammonia gradient to give the title compound as a white solid (0.971 g, 49%). 1 H NMR (400 MHz, DMSO-d6) δ 8.09-8.00 (m, 2H), 7.87 (br s, 1H), 7.75 (s, 1H), 7.35-7.25 (m, 2H), 4.54-4.45 (m, 1H), 3.14-2.92 (m, 1H), 2.87-2.17 (m, 5H), 1.98-0.98 (m, 11H) ppm. 13 C NMR (400 MHz, DMSO-d6) δ 180.1, 165.6, 162.6 (d, J = 246.4 Hz), 154.7, 131.2 (d, J = 3.0 Hz), 128.7 (d, J = 8.4 Hz), 118.2, 115.7 (d, J = (M+1) 390.

[0170] (S)-Quinuclidin-3-yl 2-(4-(4-fluorophenyl)thiazol-2-yl)propan-2-ylcarbamate (Compound 12) To a stirred solution of ethyl 3-amino-3-thioxopropanoate (20.00 g, 135.9 mmol) in ethanol (120 mL) was added 2-bromo-4'-fluoroacetophenone (29.49 g, 135.9 mmol). The mixture was refluxed for 1 h, concentrated, and partitioned between ethyl acetate (300 mL) and aqueous NaHCO3 (400 mL). The organic layer was combined with a back-extract of the aqueous layer (ethyl acetate, 1 x 100 mL), dried (Na2SO4), and concentrated. The resulting light brown solid was purified by flash chromatography using a hexane / ethyl acetate gradient to give ethyl 2-(4-(4-fluorophenyl)-2-methylpropanoate). )thiazol-2-yl)acetate as an off-white solid (29.92 g, 83%).

[0171] A stirred, cooled (−78°C) solution of ethyl 2-(4-(4-fluorophenyl)thiazol-2-yl)acetate (10.00 g, 37.69 mmol) in THF (250 mL) was added dropwise over 15 min to a solution of potassium tert-butoxide in 1.0 M THF (136 mL, 136 mmol), followed by 18-crown-6 (1.6 mL, 7.5 mmol). After an additional 30 min at −78°C, iodomethane (8.5 mL) was added dropwise over 5 min. The reaction was stirred cold for an additional 2 h, then poured into water (450 mL) and extracted with ethyl acetate (2 × 150 mL). The combined extracts were washed with brine (1 × 200 mL), dried (NaSO), and concentrated. The resulting brown oil was purified by flash chromatography using a hexane / ethyl acetate gradient to give ethyl 2-(4-(4-fluorophenyl)thiazol-2-yl)-2-methylpropanoate as a light amber oil (8.64 g, 78%).

[0172] To a stirred solution of ethyl 2-(4-(4-fluorophenyl)thiazol-2-yl)-2-methylpropanoate (0.900 g, 3.07 mmol) in 1:1:1 THF / ethanol / water (15 mL) was added lithium hydroxide monohydrate (0.451 g, 10.7 mmol). After stirring overnight, the reaction was concentrated and redissolved in water (80 mL). The solution was washed with ether (1 × 50 mL), acidified by the addition of 1 N HCl (15 mL), and extracted with ethyl acetate (2 × 50 mL). The combined extracts were dried (NaSO) and concentrated to give 2-(4-(4-fluorophenyl)thiazol-2-yl)-2-methylpropanoic acid as a pale gold solid (0.808 g, 99%).

[0173] To a stirred, cooled (0 °C) solution of 2-(4-(4-fluorophenyl)thiazol-2-yl)-2-methylpropanoic acid (0.784 g, 2.96 mmol) in THF (25 mL) was added triethylamine (0.82 mL, 5.9 mmol), followed by isobutyl chloroformate (0.58 mL, 4.4 mmol). The reaction was stirred cold for an additional 1 h before a solution of sodium azide (0.385 g, 5.92 mmol) in water (7 mL) was added. The reaction was stirred overnight, allowing the cooling bath to slowly warm to room temperature. The mixture was then diluted with water (100 mL) and extracted with ethyl acetate (2 × 60 mL). The combined extracts were washed with aqueous NaHCO (1 × 150 mL) and brine (1 × 100 mL), dried (NaSO), and concentrated. After coevaporation with toluene (2 x 30 mL), the resulting off-white solid was taken up in toluene (25 mL) and refluxed for 4 hours. (S)-3-quinuclidinol (0.753 g, 5.92 mmol) was then added and refluxing continued for 3 hours. The reaction was concentrated, and the residue was purified by flash chromatography using a chloroform / methanol / ammonia gradient to give the title compound as a white solid (0.793 g, 69%). 1 H NMR (400 MHz, CDCl3) δ 7.90-7.81 (m, 2H), 7.32 (s, 1H), 7.14-7.05 (m, 2H), 5.76 (br s, 1H), 4.72-4.65 (m, 1H), 3.26-3.10 (m, 1H), 3.03-2.37 (m, 5H), 2.05-1.23 (m, 11H) ppm. 13C NMR (400 MHz, CDCl3) δ 177.6, 162.6 (d, J = 248.4 Hz), 154.8, 153.6, 130.8 (d, J = 3.2 Hz), 128.1 (d, J = 8.1 Hz), 115.9 (d, J = 21.7 Hz), 112.2, 71.6, 55.7, 47.4, 46.5, 29.1, 25.4, 24.7, 19.6 ppm. Purity: 100 % UPLCMS (210 nm & 254 nm); retention time 0.82 min; (M+1) 390.

[0174] Quinuclidin-3-yl (2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 13) Using general procedure F, the reaction employed 2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)-2-methylpropanoic acid (prepared as described in Example 3) and quinuclidin-3-ol to afford the title compound as a colorless glassy solid (23%). NMR data were consistent with those of Example 3. Purity: 100%, 99.1% (210 and 254 nm) UPLCMS; Retention time: 0.87 min; (M+H + )439.0.

[0175] (S)-Quinuclidin-3-yl(2-(3'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 14) 2-(3'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)-2-methylpropanoic acid was prepared by exchanging 4-(2-methoxyethoxy)phenylboronic acid for 3-(2-methoxyethoxy)phenylboronic acid and using the reaction sequence outlined in Example 3. This intermediate and quinuclidin-3-ol were reacted according to general procedure F to yield the title compound as a glassy colorless solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.63-7.31 (m, 6H), 7.24-7.10 (m, 2H), 6.92 (dd, J = 8.2, 1.9 Hz, 1H), 4.51-4.34 (m, 1H), 4.21-4.08 (m, 2H), 3.72-3.64 (m, 2H), 3.32 (s, 3H), 3.09-2.26 (m, 5H), 2.04-1.22 (m, 9H) ppm. 13 C NMR (100 MHz, DMSO-d6) δ 158.9, 154.6, 147.6, 141.5, 137.6, 129.9, 126.3, 125.2, 118.9, 113.2, 112.5, 70.4, 70.0, 66.9, 58.2, 55.4, 54.2, 46.9, 45.9, 29.4, 25.3, 24.2, 19.2 ppm. Purity: 100%, 100% (210 & 254 nm) UPLCMS; retention time: 0.91 min; 15 (M+H + ) 439.4.

[0176] Quinuclidin-3-yl (2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate (Compound 15) 2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-3-yl)-2-methylpropanoic acid was prepared by exchanging ethyl 2-(4-bromophenyl)-2-methylpropanoate for ethyl 2-(3-bromophenyl)-2-methylpropanoate and using the reaction sequence outlined in Example 3. This intermediate and quinuclidin-3-ol were reacted according to general procedure F to produce the title compound as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.62-7.20 (m, 7H), 7.03 (d, J = 8.7 Hz, 2H), 4.48-4.35 (m, 2H), 4.18-4.08 (m, 2H), 3.72-3.62 (m, 2H), 3.32 (s, 3H), 3.10-2.19 (m, 6H), 2.10-1.10 (m, 11H) ppm.13 C NMR (100 MHz, DMSO-d6) δ 158.0, 154.6, 148.8, 139.5, 133.1, 128.5, 127.7, 123.8, 123.2, 122.7, 114.8, 70.4, 69.9, 67.0, 58.2, 55.3, 54.5, 47.0, 45.9, 29.4, 25.3, 24.2, 19.2 ppm. Purity: 97.4%, 94.6% (210 & 254 nm) UPLCMS; retention time: 0.88 min; (M+H + ) 439.3.

[0177] Quinuclidin-3-yl (2-(4'-(3-methoxypropoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 16) To a stirred solution of 4-iodophenol (10.05 g, 45.68 mmol) in acetonitrile (100 mL) was added potassium carbonate (6.95 g, 50.2 mmol) and 1-chloro-3-methoxypropane (6.4 mL, 57.1 mmol). The mixture was heated to reflux overnight and then concentrated. The residue was taken up in water and extracted with ethyl acetate. The combined extracts were washed with aqueous sodium bicarbonate, dried (NaSO), and concentrated. The crude material was purified by flash chromatography on silica using a hexane / ethyl acetate eluent to give 1-iodo-4-(3-methoxypropoxy)benzene as a colorless oil (4.39 g, 33%). This intermediate and ethyl 2-methyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate were reacted according to general procedure E to produce ethyl 2-(4'-(3-methoxypropoxy)-[1,1'-biphenyl]-4-yl)-2-methylpropanoate. To a stirred solution of this compound (0.693 g, 1.94 mmol) in 1:1:1 (v / v / v) tetrahydrofuran / ethanol / water (10 mL) was added lithium hydroxide monohydrate (0.326 g, 7.77 mmol). The mixture was heated to reflux overnight and then concentrated. The residue was dissolved in water, treated with 1 N hydrochloric acid (10 mL), and extracted with ethyl acetate. The combined organic layers were washed with brine, dried (Na2SO4), and concentrated to give 2-(4'-(3- Methoxypropoxy)-[1,1'-biphenyl]-4-yl)-2-methylpropanoic acid was obtained as a waxy off-white solid (0.630 g, 99%). This intermediate and quinuclidin-3-ol were reacted according to general procedure F to produce the title compound as a glassy colorless solid (62%). 1H NMR (400 MHz, DMSO-d6) δ 7.61-7.29 (m, 7H), 7.00 (d, J= 8.8 Hz, 2H), 4.47-4.36 (m, 1H), 4.05 (t, J= 6.4 Hz, 2H), 3.48 (t, J=6.3 Hz, 2H), 3.26 (s, 3H), 3.10-2.25 (m, 6H), 2.04-1.74 (m, 4H), 1.65-1.23 (m, 9H) ppm. 13 C NMR (100 MHz, DMSO-d6) δ 158.0, 154.5, 146.7, 137.4, 132.4, 127.5, 125.7, 125.2, 114.8, 69.9, 68.5, 64.6, 57.9, 55.4, 54.2, 46.9, 46.0, 29.4, 29.0, 25.2, 24.1, 19.2 ppm. Purity: 97.7%, 98.2% (210 & 254 nm) UPLCMS; retention time: 0.96 min; (M+H + ) 453.5.

[0178] Quinuclidin-3-yl (2-(4'-(hydroxymethyl)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 17) Using General Procedure E, ethyl 2-(4'-formyl-[1,1'-biphenyl]-4-yl)-2-methylpropanoate was prepared as a pale amber solid using ethyl 2-(4-bromophenyl)-2-methylpropanoate and 4-formylphenylboronic acid in the reaction. This intermediate and quinuclidin-3-ol were reacted according to General Procedure F to produce quinuclidin-3-yl (2-(4'-formyl-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate as a foamy yellow solid. To a stirred solution of this material (0.755 g, 1.92 mmol) in 2:1 (v / v) tetrahydrofuran / ethanol (15 mL) was added sodium borohydride (0.073 g, 1.93 mmol). After 45 min, the reaction was diluted with water and extracted with chloroform. The combined extracts were dried (Na2SO4) and concentrated onto silica. Flash chromatography on silica using a chloroform / methanol / ammonia eluent gave the title compound as a white solid (0.323 g, 43%). 1 H NMR (400 MHz, DMSO-d6) δ 7.66-7.29 (m, 9H), 5.18 (t, J= 5.7 Hz, 1H), 4.53 (d, J= 5.7 Hz, 2H), 4.46-4.37 (m, 1H), 3.11-2.19 (m, 6H), 2.11-1.10 (m, 11H) ppm. 13 C NMR (100 MHz, DMSO-d6) δ 154.7, 147.3, 141.5, 138.4, 137.7, 127.0, 126.2, 126.1, 125.3, 70.0, 62.6, 55.4, 54.2, 46.9, 45.9, 29.4, 25.3, 24.2, 19.2 ppm. Purity: 97.5%, 99.1 % (210 & 254 nm) UPLCMS; retention time: 0.73 min; (M+H + ) 395.

[0179] Quinuclidin-3-yl (2-(4'-(2-hydroxyethyl)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 18) Using general procedure E, ethyl 2-(4'-(2-(benzyloxy)ethyl)-4-bromobenzene and ethyl 2-methyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate were used in the reaction to prepare ethyl 2-(4'-(2-(benzyloxy)ethyl)-[1,1'-biphenyl]-4-yl)-2-methylpropanoate as a colorless gum. To a stirred solution of this compound (1.34 g, 3.33 mmol) in 1:1:1 (v / v / v) tetrahydrofuran / ethanol / water (18 mL) was added lithium hydroxide monohydrate (0.698 g, 16.6 mmol). After heating at reflux overnight, the reaction was concentrated and partitioned between water and diethyl ether. The resulting emulsion was repeatedly extracted with 0.2 N aqueous sodium hydroxide solution (5 x 50 mL). The clear portion of the aqueous layer was removed each time. The combined aqueous layers were then treated with 1.0 N hydrochloric acid (80 mL), and the resulting suspension of a white solid was extracted with ethyl acetate. The combined organic layers were dried (Na2SO4) and concentrated to give 2-(4'-(2-(benzyloxy)ethyl)-[1,1'-biphenyl]-4-yl)-2-methylpropanoic acid as a white solid (1.20 g, 96%). This compound and quinuclidin-3-ol were reacted according to General Procedure F to produce quinuclidin-3-yl (2-(4'-(2-benzyloxyethyl)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate. To a stirred solution of this material (0.435 g, 0.806 mmol) in methanol was added 1. 0N Hydrochloric acid (1 mL) and 10% palladium on carbon (50% water; 0.087 g) were added. The mixture was cycled between vacuum and nitrogen purge several times, finally evacuating and refilling with hydrogen. After 1.25 h, the reaction was filtered through Celite and concentrated. The residue was taken up in aqueous sodium carbonate and extracted with 4:1 (v / v) chloroform / isopropanol. The combined extracts were dried (Na2SO4) and concentrated onto silica. Flash chromatography on silica using a chloroform / methanol / ammonia gradient afforded the purified title compound as a colorless solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.85-7.63 (m, 1H), 7.63-7.19 (m, 8H), 4.78-4.62 (m, 2H), 3.71-2.78 (m, 8H), 2.76 (t, J= 6.8 Hz, 2H), 2.26-1.96 (m, 2H), 1.96-1.40 (m, 9H) ppm. 13 C NMR (100 MHz, DMSO-d6) δ 153.8, 146.8, 138.7, 137.9, 137.6, 129.4, 126.3, 126.1, 125.3, 66.2, 62.1, 54.4, 52.8, 45.4, 44.5, 38.6, 29.5, 29.2, 24.0, 19.9, 16.6 ppm. Purity: 100%, 100% (210 & 254 nm) UPLCMS; retention time: 0.75 min; (M+H + ) 409.

[0180] Quinuclidin-3-yl(2-(2-(4-(3-methoxypropoxy)phenyl)thiazol-4-yl)propan-2-yl)carbamate (Compound 19) To a stirred suspension of 4-methoxythiobenzamide (9.99 g, 59.7 mmol) in ethanol (75 mL) was added ethyl 4-chloroacetoacetate (8.1 mL, 60 mmol). The mixture was heated to reflux for 4 hours, then cooled, and additional ethyl 4-chloroacetoacetate (0.81 mL, 6.0 mmol) was added and returned to reflux. After heating for an additional 4 hours, the reaction was concentrated and partitioned between ethyl acetate and aqueous sodium bicarbonate. The organic layer was combined with additional ethyl acetate extracts, dried (Na2SO4), and concentrated. The crude product was purified by flash chromatography on silica using a hexane / ethyl acetate gradient to give ethyl 2-(2-(4-methoxyphenyl)thiazol-4-yl)acetate as a light amber oil (14.51 g, 87%). To a stirred solution of this compound (14.48 g, 52.2 mmol) in N,N-dimethylformamide (125 mL) was added sodium hydride (60% dispersion in mineral oil; 6.27 g, 157 mmol) in portions over 15 minutes. The resulting red suspension was cooled (0 °C) and treated dropwise with iodomethane (9.80 mL, 157 mmol) over 10 minutes. The cooling bath was removed and the reaction was stirred for 4 hours, then concentrated, and the residue was partitioned between ethyl acetate and water. The organic layer was washed twice more with water, dried (Na2SO4), and concentrated. The residue was purified by flash chromatography on silica using a hexane / ethyl acetate gradient to give ethyl 2-(2-(4-methoxyphenyl)thiazol-4-yl)-2-methylpropanoate as a pale amber oil (14.12 g, 89%). To a stirred solution of this intermediate (14.12 g, 46.24 mmol) in methylene chloride (250 mL) was added boron tribromide (11.0 mL, 116 mmol) dropwise over 5 minutes. After stirring overnight, the reaction was quenched by the slow addition of methanol (ca. 20 mL) and then concentrated. The residue was taken up in methanol (250 mL) and concentrated sulfuric acid (7.0 mL). The stirred solution was heated to reflux for 2 hours, concentrated, and partitioned between ethyl acetate and aqueous sodium bicarbonate.The organic layer was combined with the second ethyl acetate extract of the aqueous layer, dried (NaSO), and concentrated to give methyl 2-(2-(4-hydroxyphenyl)thiazol-4-yl)-2-methylpropanoate as a white solid (12.56 g, 98%). To a stirred solution of 1-bromo-3-methoxypropane (1.66 g, 10.8 mmol) in acetone (30 mL) was added the phenol intermediate (2.00 g, 7.21 mmol) and potassium carbonate (1.25 g, 9.04 mmol). The mixture was heated to reflux overnight, filtered, and concentrated. The residue was purified by flash chromatography on silica using a hexane / ethyl acetate gradient to give methyl 2-(2-(4-(3-methoxypropoxy)phenyl)thiazol-4-yl)-2-methylpropanoate as a faint amber gum (2.47 g, 98%). To a stirred solution of this compound (2.45 g, 7.01 mmol) in 1:1:1 (v / v / v) tetrahydrofuran / ethanol / water (45 mL) was added lithium hydroxide monohydrate (1.47 g, 35.0 mmol). After stirring overnight, the reaction was concentrated and washed with water and diethyl ether. The mixture was partitioned between 1.0 N hydrochloric acid (40 mL) and extracted with ethyl acetate. The combined extracts were dried (NaSO) and concentrated to give 2-(2-(4-(3-methoxypropoxy)phenyl)thiazol-4-yl)-2-methylpropanoic acid as a white solid (2.19 g, 4093%). This compound and quinuclidin-3-ol were reacted according to general procedure F to produce the title compound as a soft, faintly amber solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 8.9 Hz, 2H), 7.36 (br s, 1H), 7.24 (br s, 1H), 7.03 (d, J = 8.9 Hz, 2H), 4.49-4.41 (m, 1H), 4.07 (t, J = 6.4 Hz, 2H), 3.48 (t, J = 6.4 Hz, 2H), 3.26 (s, 3H), 3.09-2.26 (m, 6H), 2.02-1.91 (m, 2H), 1.91-1.03 (m, 11H) ppm.13 C NMR (100 MHz, DMSO-d6) δ165.8, 162.4, 160.0, 154.6, 127.5, 126.1, 114.9, 112.1, 70.1, 68.4, 64.8, 57.9, 55.4, 53.5, 46.9, 45.9, 28.9, 28.3, 25.2, 24.2, 19.2 ppm. Purity: 100%, 100% (210 & 254 nm) UPLCMS; retention time: 0.87 min; (M+H + ) 460.

[0181] Quinuclidin-3-yl(2-(2-(4-(2-methoxyethoxy)phenyl)thiazol-4-yl)propan-2-yl)carbamate (Compound 20) To a stirred solution of 2-bromoethyl methyl ether (1.88 g, 13.5 mmol) in acetone was added methyl 2-(2-(4-hydroxyphenyl)thiazol-4-yl)-2-methylpropanoate (prepared as described in Example 19, 2.00 g, 7.21 mmol) and potassium carbonate (1.56 g, 11.3 mmol). After heating at reflux overnight, the mixture was treated with additional 2-bromoethyl methyl ether (1.88 g, 13.5 mmol) and potassium carbonate (1.56 g, 11.3 mmol). The reaction was heated at reflux for two nights, filtered, and concentrated. The residue was purified by flash chromatography on silica using a hexane / ethyl acetate gradient to give methyl 2-(2-(4-(2-methoxyethoxy)phenyl)thiazol-4-yl)-2-methylpropanoate as a white solid (2.71 g, 90%). To a stirred solution of this compound (2.71 g, 8.08 mmol) in 1:1:1 (v / v / v) tetrahydrofuran / ethanol / water (50 mL) was added lithium hydroxide monohydrate (1.70 g, 40.5 mmol). After stirring overnight, the reaction was concentrated and partitioned between water and diethyl ether. The aqueous layer was treated with 1.0 N hydrochloric acid (41 mL) and extracted with ethyl acetate. The combined extracts were dried (NaSO) and concentrated to give 2-(2-(4-(2-methoxyethoxy)phenyl)thiazol-4-yl)-2-methylpropanoic acid as a white solid (2.57 g, 99%). This compound and quinuclidin-3-ol were reacted according to general procedure F to produce the title compound as a light amber solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 8.8 Hz, 2H), 7.36 (br s, 1H), 7.24 (br s, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.49-4.41 (m, 1H), 4.19-4.12 (m, 2H), 3.71-3.65 (m, 2H), 3.32 (s, 3H), 3.11-2.87 (m, 1H), 2.86-2.19 (m, 5H), 1.92-1.16 (m, 11H) ppm. 13C NMR (100 MHz, DMSO-d6) δ 165.7, 162.9, 159.9, 154.6, 127.5, 126.2, 114.9, 112.2, 70.3, 70.1, 67.1, 58.2, 55.4, 53.5, 46.9, 45.9, 28.3, 25.2, 24.3, 19.2 ppm. Purity: 100%, 100% (210 & 254 nm) UPLCMS; retention time: 0.85 min; (M+H + ) 446.

[0182] Quinuclidin-3-yl 2-(5-(4-(2-methoxyethoxy)phenyl)pyridin-2-yl)propan-2-ylcarbamate (Compound 21) 5-(4-(2-Methoxyethoxy)phenyl)picolinonitrile was prepared using general procedure E, employing 5-bromopicolinonitrile and 2-(4-(2-methoxyethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the reaction. Cerium trichloride (8.05 g, 21.6 mmol) was placed in a flask and dried by heating (170 °C) under vacuum for 3 hours. The solid was placed in tetrahydrofuran (20 mL) and stirred vigorously for 30 minutes. The suspension was cooled to -78 °C and treated dropwise with a 3.0 M solution of methyllithium in diethyl ether (7.2 mL, 21.6 mmol). After the addition, the reaction was stirred at -78 °C for 1 hour, then the resulting solution was dissolved in tetrahydrofuran (20 mL). A solution of the above aryl borate (1.83 g, 7.20 mmol) was added. The mixture was maintained at −78° C. for 2 hours and then warmed to room temperature. At this time, the reaction was quenched by the addition of aqueous ammonium hydroxide (10 mL) and filtered through a Celite plug. The filtrate was extracted with ethyl acetate, and the combined extracts were washed with brine, dried (NaSO), and concentrated. The residue was purified by flash chromatography on silica gel using ethyl acetate as eluent to afford 2-(5-(4-(2-methoxyethoxy)phenyl)pyridin-2-yl)propan-2-amine as a yellow solid (0.800 g, 39%). To a stirred suspension of this intermediate (0.500 g, 1.75 mmol) in water (10 mL) and concentrated hydrochloric acid (0.44 mL) was added toluene (10 mL). The mixture was cooled (0°C) and simultaneously treated with a solution of triphosgene (0.776 g, 2.62 mmol) in toluene (10 mL) and sodium bicarbonate (2.2 g, 26 mmol) in water (20 mL) over 1 h. After the addition, the reaction was stirred for an additional 30 min, then the top toluene layer was removed and dried (NaSO). Concurrently, a stirred solution of quinuclidin-3-ol (0.445 g, 3.64 mmol) in tetrahydrofuran (10 mL) was treated with sodium hydride (60% dispersion in mineral oil; 0.154 g, 3.85 mmol). This mixture was stirred for 5 min and then added to a solution of the crude isocyanate in toluene. The reaction was stirred for 10 min, quenched by the addition of brine (5 mL), and extracted with ethyl acetate. The combined extracts were dried (NaSO) and concentrated. The residue was purified by reverse phase silica flash chromatography to give the title compound as a pale yellow solid (0.100 g, 13%). 1H NMR (500 MHz, CDCl3) δ 8.70-8.70 (d, J = 2.0 Hz, 1H), 7.83-7.81 (m, 1H), 7.49-7.47 (d, J = 9.0 Hz, 2H), 7.45-7.43 (d, J = 8.0 Hz, 1H), 7.03-7.01 (d, J = 8.5 Hz, 2H), 6.63 (br s, 1H), 4.68-4.66 (m, 1H), 4.16 (t, J = 5.0 Hz, 2H), 3.77 (t, J = 5.0 Hz, 2H), 3.45 (s, 3H), 3.19-2.70 (m, 6H), 2.15-1.89 (m, 2H), 1.76 (s, 6H), 1.73-1.36 (m, 3H) ppm. 13 C NMR (125 MHz, CDCl3) δ 162.7, 158.9, 154.9, 145.9, 134.8, 134.3, 130.1, 128.1, 119.2, 115.2, 71.0, 70.8, 67.4, 59.2, 55.9, 55.7, 47.4, 46.5, 46.4, 27.9, 25.4, 24.6, 19.5 ppm. Purity: >99% (214 & 254 nm) LCMS; retention time: 1.32 min; (M+H + ) 440.2.

[0183] Quinuclidin-3-yl (2-(4'-(3-cyanopropoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 22) To a stirred solution of 4-bromophenol (17.1 g, 98.8 mmol) in acetonitrile (150 mL) was added 1-bromobutylnitrile (12.3 mL, 124 mmol) and potassium carbonate (15.0 g, 109 mmol). The mixture was heated to reflux overnight, cooled, and concentrated. The residue was taken up in water and extracted with ethyl acetate. The combined extracts were dried (NaSO), concentrated, and the crude material was purified by flash chromatography on silica using a hexane / ethyl acetate eluent to give 4-(4-bromophenoxy)butanenitrile as a white solid (20.8 g, 88%). To a stirred solution of this product in N,N-dimethylformamide (100 mL) was added bis(pinacolato)diboron (4.60 g, 18.1 mmol), potassium acetate (7.41 g, 75.5 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.616 g, 1.04 mmol). The mixture was heated at reflux overnight and then concentrated. The residue was taken up in ethyl acetate and washed with water and brine. The organic layer was dried (Na2SO4), concentrated, and the crude product was purified by flash chromatography on silica using a hexane / ethyl acetate eluent to give 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)butanenitrile as a white solid (3.43 g, 79%). This product and quinuclidin-3-yl (2-(4-bromophenyl)propan-2-yl)carbamate (prepared by reacting quinuclidin-3-ol and 2-(4-bromophenyl)propan-2-amine using general procedure F) were reacted according to general procedure E to yield the title compound as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.67-7.26 (m, 7H), 7.02 (d, J = 8.8 Hz, 2H), 4.50-4.33 (m, 1H), 4.08 (t, J = 6.0 Hz, 2H), 3.14-2.18 (m, 8H), 2.04 (quin, J = 6.7 Hz, 2H), 1.94-1.70 (m, 11H) ppm.13 C NMR (100 MHz, DMSO-d6) δ 157.7, 154.5, 146.8, 137.4, 132.7, 127.6, 125.7, 125.2, 120.2, 114.9, 70.0, 65.8, 55.4, 54.2, 46.9, 45.9, 29.4, 25.3, 24.7, 24.2, 19.2, 13.4 ppm. Purity: 100%, 98.9% (210 & 254 nm) UPLCMS; retention time: 0.88 min; (M+H + ) 448.6.

[0184] Quinuclidin-3-yl (2-(4'-(cyanomethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 23) Using general procedure E, the reaction employed quinuclidin-3-yl (2-(4-bromophenyl)propan-2-yl)carbamate (prepared by reacting quinuclidin-3-ol and 2-(4-bromophenyl)propan-2-amine using general procedure F) and 4-(cyanomethoxy)phenylboronic acid to prepare the title compound as a light amber solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.65 (d, J= 8.2 Hz, 2H), 7.60-7.31 (m, 5H), 7.15 (d, J = 8.9 Hz, 2H), 5.21 (s, 2H), 4.53-4.30 (m, 1H), 3.18-2.19 (m, 6H), 2.05-1.18 (m, 11H) ppm. 13 C NMR (100 MHz, DMSO-d6) δ155.8, 154.6, 147.2, 137.2, 134.4, 127.8, 126.0, 125.3, 116.7, 115.3, 70.0, 55.4, 54.2, 53.5, 46.9, 45.9, 29.4, 25.2, 24.2, 19.2 ppm. Purity: 100%, 100% (210 & 254 nm) UPLCMS; retention time: 0.85 min; (M+H + ) 420.3. Example 2

[0185] Preparation of (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate free base Step 1: Dimethylation using methyl iodide [ka] A 3N round-bottom (RB) flask was equipped with a thermometer, an addition funnel, and a nitrogen inlet. The flask was flushed with nitrogen, and potassium tert-butoxide (MW 112.21, 75.4 mmol, 8.46 g, 4.0 equiv., white powder) was weighed and added to the flask via a powder funnel, followed by THF (60 mL). A cloudy solution resulted when most of the potassium tert-butoxide dissolved. This mixture was cooled to 0–2 °C (internal temperature) in an ice-water bath. In a separate flask, the starting ester (MW 265.3, 18.85 mmol, 5.0 g, 1.0 equiv.) was dissolved in THF (18 mL + 2 mL rinse) and transferred to the addition funnel. This solution was added dropwise to the cooled mixture over 25–30 min, maintaining the internal temperature below 5 °C during the addition. The reaction mixture was cooled back to 0–2 °C. In a separate flask, a solution of methyl iodide (MW 141.94, 47.13 mmol, 6.7 g, 2.5 equiv.) in THF (6 mL) was prepared and transferred to the addition funnel. The flask containing the methyl iodide solution was then rinsed with THF (1.5 mL) and then transferred to the addition funnel already containing the clear, colorless solution of methyl iodide in THF. This solution was carefully added dropwise to the dark brown reaction mixture over 30-40 minutes, maintaining the internal temperature below 10°C throughout the addition. After the addition was complete, the slightly cloudy mixture was stirred for an additional hour, during which time the internal temperature decreased to 0-5°C. After stirring at 0-5°C for 1 hour, the reaction mixture was quenched by the slow, dropwise addition of 5.0 M aqueous HCl (8 mL) over 5-7 minutes. The internal temperature was maintained below 20°C during this addition. After the addition, water (14 mL) was added, and the mixture was stirred for 2-3 minutes. Stirring was stopped. The mixture was cooled to room temperature and the two layers were separated. The two layers were then transferred to a 250 mL 1N RB flask and as much THF as possible was evaporated in vacuo to give a biphasic layer of THF / product and water. The two layers were separated. The THF solution of the product from step 1 was used in the next reaction.

[0186] Step 2: Hydrolysis of the ethyl ester using LiOH monohydrate [ka] The crude ester in THF was added to a reaction flask. Separately, LiOH.HO (MW 41.96, 75.0 mmol, 3.15 grams, 2.2 equivalents) was weighed into a 100 mL beaker equipped with a stir bar. Water (40 mL) was added, and the mixture was stirred until all solids dissolved, resulting in a clear, colorless solution. This aqueous solution was then added to a 250 mL RB flask containing a solution of the ester in tetrahydrofuran (THF). A condenser was attached to the neck of the flask, and a nitrogen inlet was attached to the top of the condenser. The mixture was heated to reflux for 16 hours. After 16 hours, heating was discontinued, and the mixture was allowed to cool to room temperature. The THF was evaporated in vacuo to yield a brown solution. An aliquot of the brown aqueous solution was analyzed by HPLC and LC / MS to confirm the completion of the hydrolysis of the ethyl ester. Water (15 mL) was added, and the aqueous basic solution was extracted with TBME (2 × 40 mL) to remove the t-butyl ester. The aqueous basic layer was cooled to 0-10 °C in an ice-water bath and acidified with concentrated HCl dropwise while stirring to a pH of approximately 1. TBME (60 mL) was added to the sticky solid in the aqueous acidic solution, and the mixture was shaken and then vigorously stirred to dissolve all of the acid in the TBME layer. The two layers were transferred to a separatory funnel, and the TBME layer was separated. The pale yellow aqueous acidic solution was re-extracted with TBME (40 mL), and the TBME layer was separated and combined with the previous TBME layer. The aqueous acidic layer was discarded. The combined TBME layers were dried over anhydrous Na2SO4, filtered, and evaporated in vacuo to remove TBME, yielding the crude acid as an orange / dark yellow oil, which solidified under high vacuum to a dull yellow solid. The crude acid was weighed and crystallized by heating it in heptane / TBME (3:1, 5 mL / g crude) to yield the acid as a yellow solid.

[0187] Step 3: Formation of Hydroxamic Acids with NHOH.HCl [ka] The carboxylic acid (MW 265.3, 18.85 mmol, 5.0 g, 1.0 equiv.) was weighed and transferred to a 25 mL 1N RB flask under nitrogen. Upon addition of THF (5.0 mL), the acid readily dissolved, yielding a clear, dark yellow to brown solution. The solution was cooled to 0-2 °C (bath temperature) in an ice bath, and N,N'-carbonyldiimidazole (CDI; MW 162.15, 20.74 mmol, 3.36 g, 1.1 equiv.) was added slowly in small portions over 10-15 min. The ice bath was removed, and the solution was stirred at room temperature for 1 h. After stirring for 1 h, The solution was then cooled again to 0–2°C (bath temperature) in an ice-water bath. Hydroxylamine hydrochloride (NH2OH.HCl; MW 69.49, 37.7 mmol, 2.62 g, 2.0 equiv.) was added slowly as a solid in small portions over 3–5 min because the addition was exothermic. After the addition was complete, water (1.0 mL) was added dropwise to the heterogeneous mixture over 2 min, and the reaction mixture was stirred in the ice-water bath at 0–10°C for 5 min. The cooling bath was removed, and the reaction mixture was stirred overnight at room temperature under nitrogen for 20–22 h. The solution became clear as all the NH2OH.HCl dissolved. After 20–22 h, an aliquot of the reaction mixture was analyzed by high-pressure liquid chromatography (HPLC). The THF was then evaporated in vacuo, and the residue was taken up in dichloromethane (120 mL) and water (60 mL). The mixture was transferred to a separatory funnel, shaken, and allowed to separate into two layers. The aqueous layer was discarded, and the dichloromethane layer was washed with 1 N hydrochloride (HCl; 60 mL). The acid layer was discarded. The dichloromethane layer was dried over anhydrous Na2SO4, filtered, and the solvent was evaporated in vacuo to give the crude hydroxamic acid as a pale yellow solid, which was dried under high vacuum overnight.

[0188] Step 3 continuation: Conversion of hydroxamic acid to cyclic intermediate (no isolation) [ka] The crude hydroxamic acid (MW 280.32, 5.1 g) was transferred to a 250 mL 1N RB flask equipped with a nitrogen inlet. A stir bar was added, followed by acetonitrile (50 mL). The solid was insoluble in acetonitrile. The yellow, heterogeneous mixture was stirred under nitrogen for 2–3 min, and CDI (MW 162.15, 20.74 mmol, 3.36 g, 1.1 equiv.) was added in one portion at room temperature. No exotherm was observed. The solid immediately dissolved, and the clear, yellow solution was stirred at room temperature for 2–2.5 h. After 2–2.5 h, an aliquot was analyzed by HPLC and LC / MS, which indicated conversion of the hydroxamic acid to the desired cyclic intermediate.

[0189] The acetonitrile was then evaporated in vacuo to give the crude cyclic intermediate as a thick reddish oil, which was taken up in toluene (60 mL) and the reddish mixture was heated to reflux for 2 h, during which time the cyclic intermediate released CO and rearranged to the isocyanate (see below).

[0190] [ka]

[0191] Step 3 continued: Conversion of isocyanate to free base [ka] The reaction mixture was cooled to 50-60°C, and (S)-(+)-quinuclidinol (MW 127.18, 28.28 mmol, 3.6 g, 1.5 equiv.) was added to the mixture as a solid in one portion. The mixture was again heated to reflux for 18 hours. After 18 hours, an aliquot was analyzed by HPLC and LC / MS, which indicated complete conversion of the isocyanate to the desired product. The reaction mixture was transferred to a separatory funnel, and toluene (25 mL) was added. The mixture was washed with water (2 × 40 mL), and the aqueous layer was separated. The combined aqueous layers were re-extracted with toluene (30 mL), and the aqueous layer was discarded. The combined toluene layers were extracted with 1 N HCl (2 × 60 mL), and the toluene layer (containing the O-acyl impurity) was discarded. The combined HCl layers were transferred to a 500 mL Erlenmeyer flask equipped with a stir bar. This stirred, clear yellow / reddish-orange solution was basified to pH 10-12 by dropwise addition of 50% w / w aqueous NaOH. The desired free base precipitated from solution as a dull yellow sticky solid, which could be captured with a stir bar. Isopropyl acetate (100 mL) was added to the mixture, and the mixture was stirred vigorously for 5 minutes, at which point the sticky solid was converted to isopropyl acetate. Stirring was stopped, and the two layers were separated. The yellow isopropyl acetate layer was separated, and the basic aqueous layer was re-extracted with isopropyl acetate (30 mL). The basic aqueous layer was discarded, and the combined isopropyl acetate layers were dried over anhydrous Na2SO4, filtered into a pre-weighed RB flask, and the solvent was evaporated in vacuo to give the crude free base as a beige to tan solid, which was dried under high vacuum overnight.

[0192] Step 3 continued: Recrystallization of crude free base The beige to tan crude free base was weighed and recrystallized from heptane / isopropyl acetate (3:1, 9.0 mL solvent / 1 g crude free base). The appropriate amount of heptane / isopropyl acetate was added to the crude free base and a stir bar, and the mixture was heated to reflux for 10 minutes. (The free base initially partially dissolved, but upon heating to reflux, it dissolved, yielding a clear, reddish-orange solution.) When a white precipitate formed, the heat source was removed, and the mixture was cooled to room temperature with stirring. After stirring at room temperature for 3-4 hours, the precipitate was filtered off using a Buchner funnel with a vacuum hose, washed with heptane (20 mL), and dried overnight on the Buchner funnel with a vacuum hose. The precipitate was transferred to a crystallizing dish and dried in a vacuum oven at 55 °C overnight. 1 H NMR (400 MHz, CDCl3) δ 8.04 - 7.83 (m, 2H), 7.20 - 6.99 (m, 3H), 5.53 (s, 1H), 4.73 - 4.55 (m, 1H), 3.18 (dd, J = 14.5, 8.4 Hz, 1H), 3.05 - 2.19 (m, 5H), 2.0 - 1.76 (m, 11H) ppm. 13 C NMR (100 MHz, CDCl3) δ 166.38, 165.02, 162.54, 162.8-155.0 (d, CF), 130.06, 128.43, 128.34, 116.01, 115.79, 112.46, 71.18, 55.70, 54.13, 47.42, 46.52, 27.94, 25.41, 24.67, 19.58 ppm. Example 3

[0193] Preparation of crystalline forms of (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate salt The crystalline salt of (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate may be formed from the free base prepared as described in Example 23.

[0194] For example, (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate free base (approximately 50 mmol) is dissolved in IPA (140 ml) at room temperature and filtered. The filtrate is added to a 1 L RB flask equipped with an overhead stirrer and nitrogen inlet / outlet. L-Malic acid (approximately 50 mmol) is dissolved in IPA (100 + 30 ml) at room temperature and filtered. The filtrate is added to the 1 L flask. The resulting solution is stirred under nitrogen at room temperature for 4 to 24 hours (with or without seeding). Crystals form during this time. The product is collected by filtration and washed with a small amount of IPA (30 ml). The crystalline solid is dried in a vacuum oven at 55° C. for 72 hours to yield the desired malate salt.

[0195] Other salt crystalline forms, for example acid addition salts with succinic acid or HCl, may be prepared in a similar manner. Example 4

[0196] In vitro GCS inhibition (compound 2 and analogs) Inhibition of glucosylceramide synthase activity may be measured using one or more assays. The first assay is a microsomal assay, which directly measures the conversion of ceramide to glucosylceramide by HPLC. Microsomes are the source of glucosylceramide synthase activity in the microsomal assay. The second assay is a cell-based phenotypic assay, which monitors the cell surface expression of the downstream lipid GM3 by antibody-mediated immunofluorescence. Specific protocols are provided below.

[0197] Microsomal assay for glucosylceramide synthase activity: Enzyme assay using microsomes as a source of glucosylceramide synthase activity. A fluorescent ceramide substrate is delivered to the membrane-bound enzyme as a complex with albumin. After the reaction, ceramide and glucosylceramide are separated and quantified by reverse-phase HPLC with fluorescence detection. Enzyme activity is assessed using a fluorescently labeled substrate and microsomes as a source of glucosylceramide synthase. C6-NBD-ceramide is delivered in a complex with albumin to microsomes isolated according to the procedure described below. The final concentration of C6-NBD-ceramide in the stock solution is 0.5 mM; the final concentration of BSA is 0.5 mM. Separation and quantification of the substrate and product (glucosylceramide) are achieved by reverse-phase HPLC with fluorescence detection.

[0198] Preparation of microsomes from A375 human melanoma cells; Microsomes are isolated from A375 human melanoma cells. 8 to 10 million cells are harvested by trypsinization and washed with ice-cold PBS. The cells are resuspended in ice-cold lysis buffer containing protease inhibitors. The cell lysate is sonicated on ice using a probe sonicator. After sonication, the cell lysate is separated from debris by centrifugation at 10,000 g for 10 minutes at 4°C. The supernatant is removed and clarified by an additional centrifugation at 100,000 g for 1 hour at 4°C. The pellet is then resuspended in lysis buffer, aliquoted, and stored at -80°C before use.

[0199] Glucosylceramide synthase assay To determine glucosylceramide synthase inhibition, substrates (fluorescent ceramide and UDP-glucose, 3 μM and 4 μM, respectively) at twice their Km were combined with microsomes (1:50 dilution) and incubated for 1 h at room temperature on a plate shaker in the dark. The reaction was stopped by adding 150 μL of 100 μM C8-ceramide in 50% aqueous isopropanol; 10 μL of the final mix was analyzed by HPLC (with a fluorescence detector). The mobile phase was 1% formic acid in 81% methanol / 19% water. The flow rate was 0.5 mL / min. Fluorescence was measured at λ ex = 470 nm and λ em Detection was at λ = 530 nm. Under these conditions, the retention time of NBD-C6-GluCer was approximately 1.7 min, and NBD-C6-Cer eluted from the column after approximately 2.1 min. Both peaks were resolved from each other and from the baseline and were automatically integrated by the HPLC software. The conversion rate of substrate to product was used as the readout for the inhibitor test.

[0200] GM3 Fluorescence-Linked Immunosorbent Assay (FLISA): This is a phenotypic assay that measures GM3 expression in B16 mouse melanoma or C32 human melanoma cells after treatment with a test compound. Cell surface GM3 expression is determined by antibody-mediated fluorescence.

[0201] Compounds are diluted with medium and plated in DMSO in 384-well plates. B16 and C32 cells are assayed at densities of 20,000 cells / ml and 62,500 cells / ml per well, respectively. Each titration curve contains 10 points, which are assayed in duplicate in each test run. The plates are incubated at 37°C in 5% CO2 for 48 hours and then washed once with TBS. Anti-GM3 antibody is added to each well, and the plates are then incubated for an additional hour at room temperature. The plates are then washed twice and incubated with labeled secondary antibody for an additional hour. After the final incubation, the plates are washed twice and λ ex = D640 / 20nm and λ em Fluorescence at 657 nm is detected in a fluorescence reader.

[0202] Assay Results The individual assay results for certain exemplified compounds in these assays are shown in the table below. The microsomal assay results were grouped into "GCS IC 50", which represents the concentration of compound that results in 50% inhibition of glucosylceramide synthase activity. The results of the cell-based assays are labeled "GM3B16IC" for the B16 and C32 assays, respectively. 50 " or "GM3C32IC 50 These values ​​represent the concentration of compound that results in 50% inhibition of GM3 expression on the cell surface.

[0203] [Table 2]

[0204] These comparative results demonstrate that compounds according to the present disclosure have comparable in-vitro activity to GCS inhibitors and are consequently expected to demonstrate similar in-vivo benefits. Example 5

[0205] Clinical Study of Compound 2 in GD-3 Patients A 156-week, multipart, open-label, multinational study (referred to as the LEAP or LEAP2IT trial) was initiated to evaluate the safety, tolerability, pharmacokinetics, pharmacodynamics, and exploratory efficacy of Compound 2 in combination with imiglucerase in adult patients with imiglucerase-stabilized Gaucher disease type 3. Compound 2 will be administered orally in the form of malate (L-malic acid) at a single daily dose of 15 mg / day (measured as the free base). Endpoint assessments will include safety, CSF biomarkers, pharmacokinetics / pharmacodynamics, systemic disease, and neuroimaging and neurological function (CNS / neurological findings).

[0206] At least 3 years of ERT and at least 6 months on a stable monthly dose prior to enrollment Patients aged 18 years or older with a clinical diagnosis of GD3 and documented acid beta-glucosidase deficiency who had previously received treatment with imiglucerase (Cerezyme) for some time were included in the study. Patients were required to have met the following GD treatment goals: a hemoglobin level of ≥11.0 g / dL for women and ≥12.0 g / dL for men; a platelet count of ≥100,000 / mm3; a spleen volume less than 10 times normal (10 MN) or total splenectomy (provided splenectomy occurred ≥3 years prior to randomization); a liver volume less than 1.5 MN; and no bone crisis or symptomatic bone disease, e.g., bone pain due to osteonecrosis and / or pathological fractures within the last year. Patients must have GD3 characterized by oculomotor apraxia (supranuclear gaze paresis) characterized by horizontal saccade abnormalities.

[0207] (A) 52-week interim analysis (N=6) An interim analysis of six patients was conducted upon completion of 52 weeks of concurrent treatment with (1) imiglucerase (Cerezyme, Sanofi Genzyme) under each patient's established regimen, and (2) compound 2, administered orally at a single dose of 15 mg / day. During the study, patients were evaluated for safety and tolerability, CSF and plasma biomarkers (glucosylceramide, GL-1; glucosylsphingosine, lyso-GL1), pharmacokinetics, markers of systemic disease (spleen and liver volumes measured by magnetic resonance imaging (MRI), platelet count, and hemoglobin level), signs of interstitial lung disease (high-resolution computed tomography (CT) of the lung), and horizontal saccadic eye movements. Additionally, exploratory biomarkers were quantified in the CSF of GD3 patients: ceramide (a precursor of GL-1), chitotriosidase (CHITO), GM3, and GPNMB. The SARA scale was used to measure ataxia symptoms, the Trail Making Test was used to measure neurological symptoms, and functional MRI was used to assess neural connectivity in the brain.

[0208] At baseline, five patients had mild neurological involvement and one had moderate neurological involvement, as measured using the modified Severity Scoring Tool (mSST; see, e.g., Davies, et al., J Inherit Metab Dis. (2011) 34(5), pp 1053-1059).

[0209] Analysis of Compound 2 plasma and CSF concentrations indicates that Compound 2 crosses the blood-brain barrier effectively in all patients. However, patient 5 was noted to have approximately 50% lower concentrations of Compound 2 in plasma and CSF at 26 weeks and undetectable concentrations at 52 weeks. This is likely due to either compliance or dosing error, and as a result, the analysis is repeated without including patient 5's week 26 and week 52 data. The data support the conclusion that steady-state concentrations of Compound 2 are reached in plasma and CSF at or before week 4:

[0210] [Table 3]

[0211] [Table 4]

[0212] At 52 weeks, the data further demonstrate sustained and significant improvements in plasma and CSF biomarkers for GD-3. Across all six GD3 patients, plasma and CSF GL-1 and lyso-GL-1 concentrations were as follows:

[0213] [Table 5]

[0214] Thus, at 52 weeks, compared with baseline, the changes in plasma and CSF concentrations were as follows:

[0215] [Table 6]

[0216] Additionally, exploratory biomarkers were quantified in the CSF of GD3 patients: ceramide (a precursor of GL-1), chitotriosidase (CHITO; an enzyme known to be elevated in GD patients), GM3 (a glycosphingolipid marker known to be elevated in GD patients), and GPNMB (glycoprotein nonmetastatic melanoma protein B, reportedly a biomarker of neurological impairment in GD3). After 52 weeks of treatment, no significant changes were observed in CSF concentrations of ceramide, CHITO, or GPNMB. Four of six patients had measurable concentrations of GM3 in their CSF at baseline, and each of these patients was found to have undetectable GM3 in their CSF at weeks 4, 26, and 52.

[0217] Furthermore, at 52 weeks, five of the six patients showed improvement in ataxia. The degree of ataxia at baseline and throughout the study was assessed by the Scale for Assessment and Rating of Ataxia (SARA; Schmitz-Hubsch et al.

[2006] ), which assesses eight different attributes of cerebellar ataxia on a scale of 0 to 40. The eight attributes were gait, posture, sitting, speech disturbance, finger tracking, nose-to-finger test, rapid alternating hand movements, and heel-shin slide. The resulting SARA ataxia scores for all six patients are shown in the chart below:

[0218] [Table 7]

[0219] As shown in the table, five of the six patients had mild ataxia at baseline, with a mean cumulative SARA score of 2.8 (SD = 1.2). The most common deficit at baseline was gait disturbance. Excluding patient 5 due to his low level of exposure to Compound 2 and his essentially normal baseline ataxia score (only 0.5), four of the five patients showed improvement in ataxia at 52 weeks (mean improvement = -0.9; SD = 3.2). Patient 4 showed an increase in ataxia scoring, from a score of 3 at baseline to 7.5 at 52 weeks. It should be noted that this apparent deterioration was almost entirely due to a change in the "posture" scoring parameter (posture score = 1 at baseline and 26 weeks; score = 5 at 52 weeks), and the patient complained of pain in the left knee at the time of examination. Furthermore, the subject had an injury to his left big toe prior to examination; this injury was considered to have resolved 11 days after examination. Excluding these outlier effects in patient 4, treatment with Compound 2 resulted in a significant reduction in mean SARA scores by 26 weeks, which further improved slightly by 52 weeks.

[0220] The Trail Making Test (TMT) was used to assess cognitive function in patients. The TMT is one of the most widely used neuropsychological tests and is included in most test series. It is a diagnostic tool for assessing general intelligence and cognitive impairment (Tombaugh et al., 2004; Cavaco et al., 2013). In Part A of the TMT (TMT-A), subjects are asked to connect groups of digits in ascending order (Trail A). This task combines visual search and general visual acuity and motor processing speed. Part B (TMT-B) presents a sequence that alternates between digits and letters (Trail B). Connecting these in ascending but alternating order requires subjects to actively switch between both categories. Therefore, this task is considered to include an executive function component, as subjects must actively switch between categories and connect symbols simultaneously (MacPherson et al., 2017).

[0221] TMT-A primarily assesses perceptual and psychomotor speed. TMT-B more specifically assesses mental flexibility and shifting ability. Subtracting the TMT-A score from the TMT-B score is used to remove variance attributable to the graphomotor and visual scanning components of TMT-A. This subtracted score reflects the unique task requirements of TMT-B.

[0222] In a study of exemplar data for TMT-A and TMT-B in community-dwelling individuals aged 18-89 years (n = 911), the mean (SD) values ​​for the 18-24 year age group (n = 155) were 22.9 seconds (6.9) for TMT-A and 49 seconds (12.7) for TMT-B (Tombauch et al., 2004). In contrast, the mean times to complete Trail A and Trail B for patients in the study were 67.8 seconds (SD = 60.3) and 193.8 seconds (SD = 197.0), respectively. At baseline, the mean difference in time to complete Trail B minus Trail A was 126.0 seconds (SD = 142.9). This indicates that GD-3 patients in this study demonstrated some degree of cognitive impairment at baseline.

[0223] At 52 weeks, the mean time to complete Trail A was 56.5 seconds (SD = 55.2 seconds) and Trail B was 122.7 seconds (SD = 91.8 seconds). Four of six patients showed a decrease in the time it took to complete Trail A, and six of six patients showed a decrease in the time it took to complete Trail B. Excluding patient 5 due to the low level of exposure to Compound 2 in this patient, four of five patients showed a decrease in TMT-A and five of five patients showed a decrease in TMT-B.

[0224] At week 52, five of six patients showed a decrease in (TMT-B - TMT-A) time. Individual results are shown in the table below.

[0225] [Table 8]

[0226] At 52 weeks, the mean difference in time to complete Trail B minus Trail A was 66.2 seconds (SD=54.3). Excluding patient 5, 4 of the 5 patients showed improvement in Trail B minus Trail A, with a mean improvement of -71.4 seconds (-31.6%) (SD 99.3 seconds (37.6%)) at 52 weeks.

[0227] Neurological function was further assessed using functional magnetic resonance imaging (fMRI). Patient 2 was excluded because fMRI data were not collected at the 52-week session. Resting-state fMRI screening sessions were conducted at the baseline screening, 26-week, and 52-week visits. Connectivity assessments from four subjects (patients 1, 3, 4, and 5) were entered into the second-level analysis as the "adherent" group. Patient 5 was isolated due to potential non-compliance with study medication, as described above. Analyses were performed as described elsewhere (Smith et al., 2009).

[0228] Adherent subjects demonstrated enhanced connectivity across a more widely distributed set of brain regions than non-adherent subjects, most notably increased strength between posterior and anterior regions. At the anatomical level, adherent subjects demonstrated widespread and robust strengthening of connections between occipito-parietal structures and frontal, temporal, and limbic targets. Connectivity changes in Patient 5 were more subtle and restricted to spatially proximal structures. At the functional level, all subjects, except Patient 5, demonstrated enhanced connectivity between the default mode and medial frontal networks. This suggests that signals within these distinct networks become more coherent, allowing brain activity to communicate more effectively between cognitive reserve (posterior regions) and higher executive functions (anterior regions). Consistent reverse spatial mapping from resting-state networks (RSNs) 2 and 3 (cognitive-linguistic-orthographic and cognitive-spatial) to RSNs 8 and 9 (executive and left frontoparietal) is also evident. The spatial distribution of connectivity changes is even more focused for Patient 5, primarily reflecting overlap between medial-frontal and frontoparietal networks. Both perspectives suggest that the patient who fully adhered to the treatment protocol developed superior coherence between the posterior and anterior brain regions, thereby making the entire brain more susceptible to efficient information transfer. While evident, connectivity changes for Patient 5 appear within a narrower set of anterior brain regions, representing less overall evidence of therapeutic benefit.

[0229] The results are summarized in the table below. A spatial analysis of connectivity between different anatomical regions of the brain was performed to define correlation coefficients on the regressed voxel-wise mean intensities. The results showed that connectivity between the default mode (resting) network and the executive function network increased in patients 1, 3, 4, and 6, but decreased in patient 5.

[0230] [Table 9]

[0231] (B) 52-week interim analysis (N=11) An additional interim analysis was performed when 11 patients reached the 52-week milestone. The results of this analysis confirmed the observations made in the previous interim analysis.

[0232] The 11 patients in the second interim analysis included seven men and four women. Eight patients were homozygous for the p.Leu483Pro (also known as L444P) mutation in the beta-galactosidase gene, two were compound heterozygous for the p.Phe523Ile / p.Leu483Pro variant, and one was compound heterozygous for the p.Asp448His / p.Arg502Cys variant. Nine of the 11 patients had mild ataxia at baseline, primarily associated with gait disturbance. Two were considered to have no or very mild ataxia. All 11 patients remain on study. No major adverse events were reported; the most frequent were mild headache and back pain, which were deemed likely related to the lumbar puncture intervention for CSF sampling.

[0233] Plasma and CSF concentrations of Compound 2 were measured as described above. As previously observed, one patient (Patient 5) is an outlier, exhibiting very low plasma and CSF levels of Compound 2. The data continue to support the conclusion that steady-state concentrations of Compound 2 are reached in plasma and CSF at or before week 4:

[0234] [Table 10]

[0235] In the following chart, patient 5 has been excluded from the mean (N=10) values.

[0236] [Table 11]

[0237] One patient (Patient 1) also experienced a significant decrease in plasma and CSF concentrations of the compound from the 52-week measurement, which was traced to likely being the result of concomitant treatment with the CYP3A4 inducer rifampicin from study weeks 39 to 51. Because Compound 2 is suspected to be a CYP3A substrate, coadministration with a CYP3A inducer would be expected to decrease systemic exposure.

[0238] At week 52, the data further demonstrate sustained and significant improvements in plasma and CSF biomarkers for GD-3, excluding patient 5. Initial evidence of Compound 2 exposure correlated with decreases in lyso-GL1 and GL1 in CSF and plasma. Subsequently, decreases in Compound 2 exposure levels corresponded to increases in both lyso-GL1 and GL1 in both CSF and plasma. Because patient 5's CSF lyso-GL1 concentration was above the upper limit of quantitation (ULOQ; >100 pg / mL) at week 52, CSF lyso-GL1 biomarker results were estimated using the accurate ULOQ value. Thus, in patient 5, CSF lyso-GL1 and GL1 were found to increase by approximately 313% and 37%, respectively, at week 52 compared to baseline, and plasma lyso-GL1 and GL1 levels increased by approximately 43% and 14%, respectively, at week 52 compared to baseline. The mean results for other study subjects were as follows (N=10; excluding patient 5):

[0239] [Table 12]

[0240] Thus, at 52 weeks, compared with baseline, the changes in plasma and CSF concentrations were as follows: (N=10; excluding patient 5):

[0241] [Table 13]

[0242] The resulting SARA ataxia scores for the five new patients are shown in the chart below:

[0243] [Table 14]

[0244] As shown in the table, 4 of 4 patients showed improvement in ataxia at 52 weeks, except for patient 9, who did not have ataxia at baseline. Patient 7 had a temporary improvement in ataxia scoring. showed a significant increase that resolved by week 52.

[0245] To further demonstrate improvement in ataxia, two patients will be videotaped attempting to walk in a straight line at screening and at weeks 26 and 52. Comparison of the video evidence will demonstrate that, compared to baseline, both patients exhibit a more stable, better coordinated, and faster gait, with less contact with the nearby wall for support and less side-stepping.

[0246] TMT timing for new patients is shown in the chart below.

[0247] [Table 15]

[0248] At 52 weeks, three new patients showed a small increase in (TMT-B-TMT-A) time of uncertain clinical significance, two new patients showed a decrease in (TMT-B-TMT-A) time, and one patient showed a very large decrease (92% decrease).

[0249] Neurological function was further assessed in new patients using functional magnetic resonance imaging (fMRI), as described above. fMRI results continue to demonstrate that patients adequately exposed to Compound 2 developed greater coherence between the posterior and anterior brain regions, resulting in the entire brain being more susceptible to efficient information transfer. While connectivity changes were evident in a single patient (Patient 5) with insufficient Compound 2 exposure, they emerged within a narrower set of anterior brain regions, representing less overall evidence of therapeutic benefit. This enhanced connectivity was seen in regions associated with executive function. Resting-state functional MRI demonstrated enhanced connectivity between the default mode and medial frontal networks, suggesting that signals within these distinct networks became more coherent, allowing brain activity to more effectively communicate between cognitive reserve (posterior regions) and higher executive function (anterior regions).

[0250] In particular, the results show enhanced connectivity between RSNs 1, 2, and 3 (perceptual-visual, cognitive-linguistic-orthographic, and cognitive-spatial) and RSNs 6, 7, and 8 (sensorimotor, auditory, and executive control). At the anatomical level, there is widespread and robust strengthening of connections between occipito-parietal structures and frontal, temporal, and limbic targets. The data suggest increased functional connectivity, most prominent within the sensory, motor, and cerebellar networks thought to be disrupted in Gaucher disease.

[0251] For the change in correlation coefficient between baseline and week 52 (patient 2 was excluded due to missing data, as noted above), for all patients in RSNs 3 and 6 (RSNs 4 and 8 were not included), the results are summarized in the table below.

[0252] [Table 16]

[0253] Statistical analysis of SARA and TMT results compared with GL-1 concentrations in CSF at 52 weeks was found to provide good therapeutic correlation. For SARA results, where lower scores indicate improved treatment, 8 of 11 patients showed a positive correlation between the decrease in CSF GL-1 (ng / mL) and SARA score (-5 to 5). For TMT, where shorter times (trail B minus trail A) indicate improved treatment, 6 of 11 patients showed a positive correlation between the decrease in CSF GL-1 (ng / mL) and TMT time (seconds).

[0254] (C) Additional 52-week interim analysis (N=9) Neurological function was further assessed using volumetric magnetic resonance imaging (fMRI). vMRI data were collected at baseline screening sessions and after 52 weeks of treatment regimen for 8 patients ("Group A") and an additional isolated patient (Patient 5). Group A corresponds to the 11 patients described above, excluding Patient 5 and two other patients with insufficient analyzable vMRI data. Patient 5 was isolated due to plasma and CSF concentrations of Compound 2 lower than the LLOQ after 52 weeks, suggesting failure to comply with the treatment regimen.

[0255] vMRI data were acquired and subsequently analyzed using an anatomical parcellation and tensor-based morphometry (TBM) analysis cycle with FreeSurfer. FreeSurfer is an open-source software package for the analysis and visualization of structural and functional neuroimaging data developed by the Laboratory for Computational Neuroimaging. TBM analysis (also known as Jacobian integration) consists of estimating the volumetric changes captured within the deformation field resulting from the application of a symmetric deformable registration technique between a pair of MR scans (baseline and follow-up) using a nonlinear symmetric log-demons deformation technique (a symmetric process) with a robust cross-correlation metric to ensure transformation inversion. The deformation field is then analyzed by calculating the determinant of the Jacobian matrix, a measure of local volumetric change. Integration of the determinant over the region of interest provides an estimate of the rate of volume change in this brain region over time.

[0256] The overall pipeline takes baseline (BL) and follow-up (FU) images as input and consists of the following steps: 1. Preprocessing and reformatting; 2. Baseline (BL) and follow-up (FU) segmentation using FreeSurfer; 3. Multi-resolution rigid and affine registration of FU and BL to midspace; 4. Symmetric variable nonlinear registration between FU and BL; 5. Computation of Jacobian images; 6. Calculation of voxel-wise volume changes; 7. Integrating the volume change per area; and 8. Output of changes in the areas of interest specified in the study.

[0257] Thus, TBM is an image analysis technique that identifies regional structural differences from the gradient of a nonlinear deformation field that aligns an image to a general anatomical template. TBM is a well-known tool for analyzing vMRI data of the brain, and further discussion of the application of related techniques can be found, for example, in John Ashburner and Karl J Friston, Human Brain Function, Second edition, Academic Press 2004, Section 1, Chapter 6.3, pages 8 to 13, ISBN: 9780080472959; Moo K Chung, Computational Neuroanatomy, World Scientific 2012, Chapter 3, Pages 49 to 68, ISBN: 9789814472814. doi.org / 10.1142 / 8036; and Thomson et al., Ann NY Acad Sci. 2007 February; 1097: 183-214. doi: 10.1196 / annals.1379.017.

[0258] vMRI data were collected on the entire brain tissue, allowing analysis to quantify volumetric changes in individual regions of brain tissue. Results from Group A demonstrate that after 52 weeks of the treatment regimen, brain tissue volume increased in numerous individual brain regions, resulting in an increase in total brain volume. Brain regions with increased volume were also found to overlap with regions showing increased neuronal connectivity, as measured by fMRI. Conversely, results from Patient 5 demonstrate evidence of brain atrophy and a decrease in total brain volume.

[0259] After 52 weeks of treatment, Group A demonstrated increases in mean volume in at least the following brain regions: right nucleus accumbens region, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine lobe, right amygdala, left cuneus, and left lingual gyrus, as well as increases in mean whole brain volume. These results are shown in the table below.

[0260] [Table 17]

[0261] Also, as can be seen in Figure 1, while Group A experienced an overall increase in whole brain tissue volume as a result of the treatment regimen, Patient 5, on the other hand, experienced a decrease in whole brain volume. Patient 5, who was not successful in receiving the treatment regimen, showed a relatively severe decrease in whole brain volume, as well as volume loss in most of the individual brain regions mentioned above. These results indicate that the treatment regimen not only increased brain tissue volume in Group A, as shown in Patient 5, but also prevented and / or delayed potential loss of brain tissue volume in Group A as a result of GD3 disease progression. Example 6

[0262] Pharmacokinetics of Compound 2 in Healthy Human Volunteers A two-part Phase 1 clinical study was conducted to evaluate the pharmacokinetics, pharmacodynamics, safety, and tolerability of Compound 2, also known as benglustat, in healthy human volunteers in the presence and absence of food.

[0263] Research 1 Study 1 was a two-part, single-center trial in healthy adult male volunteers. Part 1 was a double-blind, randomized, placebo-controlled, ascending-dose, single-dose study of Compound 2 for safety, tolerability, and PK. Part 2 was an open-label, single-cohort, randomized, two-sequence, two-period, two-treatment, crossover study of Compound 2 for PK with and without a high-fat meal.

[0264] In Part 1 of the study, 55 healthy men (placebo, n=14; 2, 5, 15, 25, 50, and 100 mg doses, n=6 each; 150 mg dose, n=5) were enrolled and randomized. Eight healthy men participated in Part 2.

[0265] In Part 1, subjects were randomized to receive 2, 5, 15, 25, 50, 100, or 150 mg of Compound 2 (expressed in the L-malate form) or a matching placebo on the first morning after at least 10 hours of fasting. In Part 2, subjects were randomized to receive a single oral dose of 5 mg of Compound 2 administered either in the fasted state (at least 10 hours before and 4 hours after administration) or 30 minutes after a standardized high-fat breakfast (approximately 815 kcal). After a 7-day washout period, participants were crossed over to the other condition.

[0266] In Study 1, Part 1, blood was sampled for Compound 2 plasma concentrations at the time of study drug administration (0 hours) and at 0.5, 1, 2, 3, 4, 5, 6, 8, 10, 12, 16, 24, 48, 72, and 96 hours post-dose. Urine samples were collected and analyzed for Compound 2 concentrations beginning 2 hours before and continuing through 48 hours after study drug administration.

[0267] In Study 1, Part 2, blood was sampled for plasma concentrations of Compound 2 at 0, 0.5, 1, 2, 3, 4, 5, 6, 8, 10, 12, 16, 24, and 48 hours after dosing.

[0268] From Part 1, the maximum plasma concentrations (C) of Compound 2 were measured after single oral administration of doses of 2 to 150 mg. max ) occurred at a median time of 3 to 5.5 hours before the plasma concentration began to decline exponentially, and the geometric mean t 1 / 2 The exposure increased approximately dose-proportionally across the dose range: a 75-fold dose increase resulted in a geometric mean C max , AUC last , and AUC inf The PK results were 97.3-, 89.2-, and 85.9-fold increases in PK values, respectively. The PK results are shown in the table below (AUC = area under the time-concentration curve where a measurable concentration persists or is extrapolated to infinity; t 1 / 2 = terminal half-life; CL / F = apparent total clearance from plasma; CV = coefficient of variation; SD = standard deviation; tmax =C max time to release; Vss / F = apparent volume of distribution at steady state):

[0269] [Table 18]

[0270] Part 2 showed that a 5 mg dose administered with a high-fat meal had no effect on Compound 2 exposure compared to fasting conditions. Median t max was 6.00 h whether fed or fasted. The geometric mean fed / fasted ratio was C max and AUC last The values ​​were 0.92 and 0.91 for , , and , respectively. Within-subject variability (i.e., fed vs. fasted) accounted for less than half of the total subject variability.

[0271] Research 2 Study 2 was a single-center, double-blind, randomized, placebo-controlled, ascending-dose study of the safety, tolerability, PK, and pharmacodynamics of Compound 2 in healthy adult male and female volunteers.

[0272] The study enrolled and randomized 36 healthy adults (19 men and 17 women) (n = 9 per group) to receive Compound 2 (provided in the form of 5-mg capsules of the L-malate salt) or placebo once daily for 14 days after at least a 10-hour fast.

[0273] Blood was sampled for Compound 2 plasma concentrations as follows: Day 1, 0, 0.5, 1, 2, 3, 4, 5, 6, 8, 10, 12, and 16 hours post-dose; Days 2-5, 8, 11, and 13, 0 hours post-dose; Day 14, 0.5, 1, 2, 3, 4, 5, 6, 8, 10, and 12 hours post-dose; Days 15-17, 24, 48, and 72 hours post-dose on Day 14, respectively. Urine samples were collected and analyzed for Compound 2 concentrations 0-24 hours post-dose on Day 1 (0 hours post-dose) and serially on Day 14. Pharmacodynamic endpoints were analyzed. Time points (plasma GL-1, GL-3, and GM3 concentrations) were assessed at 0 h post-dose on days 1–5, 8, 11, 13, and 14; and 24 h post-dose on day 15.

[0274] In subjects receiving 5, 10, or 20 mg of Compound 2 once daily for 14 days, plasma C max C occurred at a median time of 2 to 5 hours after dosing on days 1 and 14. trough Compound 2 exposure increased approximately dose-proportionally over the 5-20 mg dose range: this four-fold dose increase resulted in a geometric mean C value of 0.01 at day 14. max and AUC 0-24 There was a 3.76- and 3.69-fold increase in values, respectively. The PK results from Study 2 are summarized in the table below:

[0275] [Table 19]

[0276] After 14 days of once-daily administration of Compound 2, the 24-hour urinary excretion rate of unchanged compound (mean fe 0-24 ) ranged between 26.3% and 33.1% and had no apparent dose-related association. R(0-24) ranged between 1.49 L / h and 2.07 L / h, approximately 3.18-3.86 times lower than the observed plasma CL / F.

[0277] Plasma GL-1, GL-3, and GM3 in placebo recipients remained similar across baseline, while across the three Compound 2 dose groups, plasma GL-1 and GM3 levels decreased from baseline in a time- and dose-dependent manner as shown in the following table (point estimates of treatment ratios for glucosylceramide (GL-1), globotriaosylceramide (GL-3), and GM3 ganglioside (GM3) at day 15 of the repeated ascending dose study):

[0278] [Table 20]

[0279] The maximum sustained effect for GL-1 occurred on Day 11 in the 5 and 10 mg groups and by Day 8 in the 20 mg group. The mean calculated GL-1 reduction from baseline on Day 15 was 41.9%, 69.6%, and 74.6% in the 5, 10, and 20 mg groups, respectively. GL-1 values ​​were below the lower limit of quantification (LLOQ) at baseline in one 5 mg Compound 2 recipient and on Day 15 in three, five, and nine subjects in the 5, 10, and 20 mg groups, respectively.

[0280] The greatest sustained GM3 reduction occurred across all dose groups of Compound 2, beginning on Day 13. Mean plasma GM3 levels on Day 15 were 42.7%, 49.4%, and 57.8% of baseline for the 5, 10, and 20 mg dose groups, respectively. GM3 was below the LLOQ on Day 15 in one and two subjects in the 10 and 20 mg dose groups, respectively.

[0281] Plasma GL-3 also decreased over time in all Compound 2 dose groups, but baseline GL-3 values ​​were variable and low compared to the LLOQ, limiting the calculated mean decrease in GL-3. In the placebo, 5, 10, and 20 mg dose groups, GL-3 values ​​were below the LLOQ in 1, 3, 1, and 6 subjects, respectively, at baseline, and in 4, 9, 7, and 9 subjects, respectively, at Day 15.

[0282] C of Compound 2 in the 5, 10, and 20 mg dose groups trough The mean estimated plasma GL-1 reductions from baseline (19.0, 47.5, and 69.9 ng / mL, respectively) (90% CI) were 67.0% (54.4-79.7%), 74.4% (63.7-85.2%), and 76.3% (64.8-87.8%), respectively.

[0283] conclusion In these studies, exposure to Compound 2 (C max The AUC and AUC were approximately dose-proportional when administered over a 14-day period at doses ranging from 2 to 150 mg as a single dose or from 5 to 20 mg as repeated once-daily doses. A high-fat meal, compared with fasting, had no effect on exposure in subjects receiving a single 5 mg dose. At repeated once-daily doses of 5 to 20 mg, steady-state was reached within 5 days; neither age nor gender affected accumulation. Pharmacodynamically, repeated once-daily doses of Compound 2 reduced plasma concentrations of GL-1 and GM3 in a time- and dose-dependent manner, consistent with Compound 2-mediated GCS inhibition, although baseline levels of GL-3 were too low to be useful as a pharmacodynamic biomarker. The dose-dependent reduction in GL-1 supported the intended mechanism of action of Compound 2: inhibition of GL-1 formation from ceramide by GCS.

[0284] In all studies, the safety profile, including serious adverse events [SAEs], ECG monitoring, clinical laboratory values, and physical examinations, was assessed by treatment-emergent adverse event (TEAE) monitoring for 10 days after the last dose of study medication. There were no deaths, SAEs, severe TEAEs, or TEAEs leading to study discontinuation in either study.

[0285] No clinically relevant blood or biochemical abnormalities were reported in any study. Vital signs showed no relevant changes from baseline in any study. ECG parameters showed no relevant changes in ascending single-dose and food effect studies; in the multiple ascending-dose study, ECG parameters did not change statistically significantly from mean baseline vs. placebo in Compound 2 recipients at any dose. While the present invention has been described in conjunction with the above embodiments, it should be understood that the foregoing descriptions and examples are intended to be illustrative and not limiting of the scope of the invention. Other aspects, advantages, and modifications within the scope of the invention will be apparent to those skilled in the art to which the present invention pertains.

[0286] Furthermore, when features or aspects of the invention are described in terms of a Markush group, one skilled in the art will recognize that the invention is also described with respect to any individual member or subgroup of members of the Markush group.

[0287] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety to the same extent as if each was individually incorporated by reference. In case of conflict, the present specification, including definitions, will control.

Claims

1. A method for enhancing neuronal connectivity in the brain in a subject, such as a patient, in need thereof, comprising administering to the subject an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or prodrug thereof. 【Chemistry 1】 (In the formula: R １ is hydrogen, halogen (e.g., fluorine), cyano, nitro, hydroxy, thio, amino, C １～６ - alkyl (e.g., methyl or ethyl), C ２～６ -alkenyl, C ２～６ -alkynyl, C １～６ -Alkyloxy, C ２～６ -alkenyloxy, and C ２～６ -alkynyloxy, wherein said alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy, or alkynyloxy is optionally substituted with one or more (e.g., 1, 2, or 3) groups selected from halogen, cyano, nitro, hydroxy, thio, and amino; R ２ and R ３ is a C optionally substituted by one or more (e.g., 1, 2, or 3) halogens; １～３ -alkyl, or R ２ and R ３ together form a cyclopropyl or cyclobutyl group optionally substituted by one or more (e.g., one or two) halogens; R ４ , R ５ and R ６ is hydrogen, halogen, nitro, hydroxy, thio, amino, C １～６ - alkyl, and C １～６ -alkyloxy, wherein said alkyl or alkyloxy is selected from halogen, hydroxy, cyano, and C １～６ - optionally substituted by one or more (e.g., 1, 2, or 3) groups selected from alkyloxy; A is halogen, hydroxy, thio, amino, nitro, C １～６ Alkoxy and C １～６ and a 5- or 6-membered aryl or heteroaryl group (e.g., phenyl or thiazolyl) optionally substituted with 1, 2, or 3 groups independently selected from alkyl.

2. 2. The method of claim 1, wherein the compound is selected from quinuclidin-3-yl (2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate; (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate; and (S)-quinuclidin-3-yl (2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate; and pharmaceutically acceptable salts and prodrugs thereof.

3. 2. The method of claim 1, wherein the compound is quinuclidin-3-yl (2-(4'-fluoro-[1,1'-biphenyl]-3-yl)propan-2-yl)carbamate.

4. The compound of claim 1, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate. method.

5. 2. The method of claim 1, wherein the compound is (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate in the form of a malate salt.

6. The method of any one of claims 1 to 5, wherein the subject has Gaucher disease type 3.

7. The method of any one of claims 1 to 6, wherein the compound, or a pharmaceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a route that is not parenteral.

8. 8. The method of claim 7, wherein the compound, or a pharmaceutically acceptable salt or prodrug thereof, is administered orally.

9. 9. The method of any one of claims 1-8, wherein the subject is undergoing concurrent treatment with an enzyme replacement therapy (ERT), e.g., with glucocerebrosidase (e.g., imiglucerase, velaglucerase, or taliglucerase).

10. 10. The method of any one of claims 1 to 9, wherein the subject is administered a daily dose of about 1 mg to about 50 mg of the compound, for example, 5 to 50 mg, or 10 to 40 mg, or 10 to 30 mg, or 10 to 20 mg, or 20 to 30 mg, or 30 to 40 mg, or 40 to 50 mg, or 5 to 25 mg, or 20 to 50 mg, or 5 to 15 mg, or 15 to 30 mg, or about 15 mg of the compound.

11. 11. The method of any one of claims 1 to 10, wherein the subject is administered a single daily dose of 15 mg (measured as the amount of free base) of (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate in the form of the malate salt.

12. 12. The method of any one of claims 1 to 11, wherein the compound is not administered simultaneously with a strong or moderate inducer of CYP3A, such as rifampin, phenobarbital, or efavirenz.

13. 13. The method of any one of claims 1 to 12, wherein the subject is an adult or pediatric patient 12 years of age or older with Gaucher disease type 3 who has been stabilized on enzyme replacement therapy (ERT) using imiglucerase for performance status, and the subject is administered a single daily dose of 15 mg (measured as the amount of the free base) of (S)-quinuclidin-3-yl(2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate in the form of the malate salt.

14. 14. The method of any one of claims 1 to 13, which is effective to improve cognitive performance or reduce a cognitive deficit, e.g., as measured by a reduction in the time taken to complete the Trail Making Test (TMT), TMT-A and / or TMT-B, a reduction in the difference between the TMT-A time and the TMT-B time (TMT-A-TMT-B), e.g., a reduction of at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50% (e.g., a 5-20% decrease in TMT-A, and / or a 25-30% decrease in TMT-B, and / or a 25-30% decrease in [TMT-A-TMT-B]).

15. 15. The method of any one of claims 1 to 14, wherein the method results in increased blood flow in the brain (e.g., in one or more of the frontal lobe, occipital lobe, parietal lobe, or temporal lobe), e.g., as shown by fMRI imaging.

16. 16. The method of any one of claims 1 to 15, wherein the method results in increased nodal connectivity in the brain (e.g., between the posterior and anterior sides of the brain, and / or between occipital-parietal structures and frontal, temporal and / or limbic structures, as shown by fMRI imaging).

17. 17. The method of any one of claims 1 to 16, resulting in enhanced connectivity in brain regions associated with executive function.

18. 18. The method of any one of claims 1 to 17, resulting in a resting state functional network with improved connectivity between the default mode and the medial and frontal networks.

19. 19. The method of any one of claims 1 to 18, wherein the method results in enhanced connectivity between RSNs 1, 2, and 3 (perceptual-visual, cognitive-linguistic-orthographic, cognitive-spatial) and RSNs 6, 7, and 8 (sensorimotor, auditory, and executive control).

20. 20. A compound as defined in any one of claims 1 to 5, or a pharmaceutically acceptable salt or prodrug thereof, for use in a method for enhancing neuronal connectivity in the brain of a subject, such as a method as defined in any one of claims 6 to 19.

21. For example, use of a compound as defined in any one of claims 1 to 5, or a pharmaceutically acceptable salt or prodrug thereof, in the manufacture of a medicament for enhancing neuronal connectivity in the brain of a subject, as defined in any one of claims 6 to 19.

22. 10. A method for increasing brain tissue volume or preventing or slowing the loss of brain tissue volume in a subject, such as a subject in need thereof, said method comprising administering to the subject an effective amount of a compound as defined in any one of claims 1 to 5.

23. 23. The method of claim 22, wherein an increase in brain tissue volume, or prevention or delay of loss of brain tissue volume, is achieved in one or more brain regions selected from the right nucleus accumbens, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine sulcus, right amygdala, left cuneus, and left lingual gyrus.

24. 24. The method of claim 22 or claim 23, wherein the method results in an increase in brain volume in one or more brain regions associated with executive function.

25. 25. The method of any one of claims 22 to 24, wherein the increase in brain volume in one or more brain regions is accompanied by enhanced neuronal connectivity within the one or more brain regions, as shown, for example, using functional magnetic resonance imaging (fMRI).

26. 26. The method of any one of claims 22 to 25, wherein the method results in an increase in total brain tissue volume.

27. 27. The method of any one of claims 22 to 26, wherein the administration of the subject and / or the compound of formula (I), or a pharmaceutically acceptable salt or prodrug thereof, is as defined in any one of claims 6 to 13.

28. A composition according to claim 1 for use in a method as defined in any one of claims 22 to 27.

6. A compound as defined in any one of claims 1 to 5, or a pharmaceutically acceptable salt or prodrug thereof.

29. 28. Use of a compound as defined in any one of claims 1 to 5, or a pharmaceutically acceptable salt or prodrug thereof, in the manufacture of a medicament for use in a method as defined in any one of claims 22 to 27.

30. A method for monitoring the progression or regression of a neurological disorder associated with a lysosomal storage disease in a subject, wherein the subject is undergoing treatment comprising administering to the subject an effective amount of a compound as defined in any one of claims 1 to 5, the method comprising measuring the subject's brain tissue volume over a period of time during the course of treatment, for example using volumetric magnetic resonance imaging (vMRI), and assessing the degree of change in brain tissue volume over said period.

31. 31. The method of claim 30, wherein the period is from 3 to 24 months, e.g., from 3 to 12 months, from 3 to 6 months, from 6 to 24 months, from 6 to 18 months, from 6 to 12 months, from 12 to 24 months, or from 12 to 18 months.

32. 32. The method of claim 30 or claim 31, further comprising modifying the treatment by increasing the dosage of the compound of formula (I) or a pharmaceutically acceptable salt or prodrug thereof administered to the subject if there is a decrease or no increase in total brain volume observed over said period of time, and reassessing the extent of change in brain tissue volume after a further period of time over the course of modified treatment with increased dosage.

33. 32. The method of claim 30 or claim 31, further comprising modifying the treatment by increasing the dosage of the compound of formula (I) or a pharmaceutically acceptable salt or prodrug thereof administered to the subject if there is a decrease or no increase in volume in three or more of the following brain regions observed over the period: right nucleus accumbens region, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcaneus, right amygdala, left cuneus, and left lingual gyrus, and re-evaluating the degree of change in brain tissue volume after a further period over the course of the modified treatment with increased dosage.

34. 34. The method of any one of claims 30 to 33, wherein the administration of the subject and / or the compound of formula (I), or a pharmaceutically acceptable salt or prodrug thereof, is as defined in any one of claims 6 to 13.

35. A compound as defined in any one of claims 1 to 5 for use in a method as defined in any one of claims 30 to 34.

36. 35. Use of a compound as defined in any one of claims 1 to 5 in the manufacture of a medicament for treatment by a method as defined in any one of claims 30 to 34.

37. 1. A method for assessing the development of a neurological disorder associated with a lysosomal storage disease in a subject at risk of developing said neurological disorder, comprising: a) measuring the subject's brain tissue volume (e.g., using vMRI) and comparing with a reference standard to assess whether the brain tissue volume is lower than the reference standard; b) identifying the onset of said neurological disorder if the brain tissue volume identified in step (a) is lower than a reference standard; c) initiating treatment of a subject by administering to the subject an effective amount of a compound as defined in any one of claims 1 to 5 or a pharmaceutically acceptable salt or prodrug thereof. The method optionally further comprising:

38. 38. The method of claim 37, further comprising administering to a subject a compound as defined in any one of claims 1 to 5, or a pharmaceutically acceptable salt or prodrug thereof.

39. The method of claim 37 or claim 38, wherein the subject is undergoing concurrent treatment with imiglucerase.

40. 40. The method of any one of claims 37 to 39, wherein the subject has been administered an enzyme replacement therapy drug (e.g. imiglucerase, velaglucerase and / or taliglucerase) prior to commencing any treatment with a compound as defined in any one of claims 1 to 5.

41. 41. The method of any one of claims 37 to 40, wherein the subject has been receiving imiglucerase therapy (optionally at a stable dose) for at least 6 months prior to commencing any therapy with a compound as defined in any one of claims 1 to 5.

42. 42. The method of any one of claims 37 to 41, further comprising the step of transitioning the subject from an ERT therapeutic agent (e.g., imiglucerase, velaglucerase or taliglucerase) to any treatment with a compound as defined in any one of claims 1 to 5.

43. 43. The method of any one of claims 37-42, wherein the subject has a hemoglobin level of at least 11 g / dL for women and at least 12 g / dL for men; a platelet count of at least 100,000 per cubic millimeter; a spleen volume less than 10 times normal (10 MN); and / or a liver volume less than 1.5 MN.

44. 44. The method of any one of claims 37 to 43, wherein measuring the subject's brain tissue volume is by brain positron emission tomography (PET) or volumetric magnetic resonance imaging (vMRI).

45. 45. The method of any one of claims 37 to 44, wherein the subject's brain tissue volume is found to be lower than a reference standard.

46. 46. ​​The method of any one of claims 37-45, wherein comparison with a reference standard indicates that the subject has lower brain tissue volume in one or more brain regions selected from the right nucleus accumbens, left putamen, left entorhinal cortex, right putamen, right postcentral lobe, left calcarine sulcus, right amygdala, left cuneus, and left lingual gyrus.

47. 47. The method of any one of claims 37 to 46, wherein comparison with a reference standard indicates that the subject has lower brain tissue volume in one or more brain regions associated with executive function.

48. 48. The method of any one of claims 37 to 47, wherein comparison with a reference standard indicates that the subject has lower brain tissue volume in one or more brain regions assessed as having loss of neuronal connectivity, as shown, for example, using functional magnetic resonance imaging (fMRI).

49. 49. The method of any one of claims 37 to 48, wherein comparison with a reference standard indicates that the subject has a lower total brain tissue volume.

50. The brain tissue volume of the subject is measured after treatment with a compound as defined in any one of claims 1 to 5.

50. The method of any one of claims 37 to 49, wherein the brain tissue volume is measured multiple times intermittently or periodically, for example weekly, monthly, every 2, 3, 4, 6, 9, 12 months, etc., after the measurement to assess changes in brain tissue volume.

51. 51. The method of any one of claims 37 to 50, wherein the subject and / or any administration of a compound of formula (I), or a pharmaceutically acceptable salt or prodrug thereof, is as defined in any one of claims 6 to 13.

52. A compound as defined in any one of claims 1 to 5 for use in a method as defined in any one of claims 37 to 51.

53. 53. Use of a compound as defined in any one of claims 1 to 5 in the manufacture of a medicament for treatment by a method as defined in any one of claims 37 to 52.

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