Treatment of ciliary diseases using glucosylceramide synthase (GCS) inhibitors

Abstract

The present disclosure relates to a method for treating a ciliopathic disease in a subject, the method comprising administering to the subject an effective amount of a quinuclidine compound. Also disclosed are pharmaceutical compositions comprising the quinuclidine compound for use in the method.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is an international application claiming priority to and benefit of U.S. Provisional Patent Application No. 62 / 800,993, filed February 4, 2019, and U.S. Provisional Patent Application No. 62 / 851,430, filed May 22, 2019, the contents of each of which are hereby incorporated by reference in their entirety.

[0002] The present invention relates to methods for treating ciliary diseases, such as Bardet-Biedl Syndrome (BBS) and Joubert Syndrome, using the quinuclidine compounds of formula (I). [Background technology]

[0003] Ciliopathies are a group of diseases / disorders associated with genetic mutations that code for defective proteins, resulting in abnormal formation and function of cilia. Cilia are components of most types of cells in the body. Thus, abnormalities in cilia formation and function can result in a collection of features including, but not limited to, retinal degeneration, kidney disease, and brain abnormalities. Some of the diseases / disorders result from these ciliopathies, including Joubert syndrome, Meckel-Gruber syndrome, Senior-Loken syndrome, Orofacial-Digital syndrome type I, Leber congenital amaurosis, Bardet-Biedl syndrome (BBS), Amström syndrome, June asphyxiating thoracic dystrophy, Ellis-van Creveld syndrome, Sensenbrenner syndrome, primary ciliary dyskinesia (also known as Kartagener syndrome), and various other diseases and disorders.

[0004] For example, among ciliopathies, BBS represents a completely unmet clinical need, and there are currently no approved treatment options for BBS patients. BBS is a rare autosomal recessive multisystem genetic disorder with a prevalence of 1:160,000 in the United States and Northern Europe. BBS can result from mutations in at least 21 different genes, with mutations in BBS1, BBS2, and BBS10 accounting for approximately 50% of cases. The genes affected in BBS are essential for the assembly of the BBSome, a component of the basal body, which is involved in the formation, maintenance, and function of primary cilia. At the same time, primary cilia and their anchoring structure, the basal body, are essential for the proper function of many key biological signaling pathways. Loss of a properly formed BBSome leads to widespread loss of cilia, which is manifested by multiple clinical features. Eight BBS proteins (BBS1, BBS2, BBS4, BBS5, BBS7, BBS8, BBS9, and BBS18) assemble to form the BBSome complex. The functions of these BBS proteins are partially overlapping, which is consistent with the phenotypic similarities observed with mutations in individual BBS genes. Studies have shown that loss of individual BBS protein functions can result in the same phenotypic deficiencies, and more than one BBS gene or protein can be targeted simultaneously to achieve the same therapeutic effect. For example, in vitro inhibition of BBS4, BBS10, and BBS12 in differentiating preadipocytes promotes adipogenesis and fat accumulation (Non-Patent Document 1; Non-Patent Document 2). Loss of BBS1 and BBS4 results in defects in the localization of certain proteins, and the olfactory epithelium fails to fully develop cilia (Non-Patent Document 3). Furthermore, loss of BBS8 results in loss of cilia in olfactory sensory neurons, mislocalization of cilia-associated proteins, and reduced response to olfactory stimuli (Non-Patent Document 4). Finally, it has been observed that deletion of BBS2 reduces adenylate cyclase III activity in the primary olfactory epithelium, the same being observed in BB1, BBS4 and BBS8 null mice.This effect is ameliorated by treatment with glucosylceramide synthase (GCS) inhibitors.

[0005] The primary features of BBS are cone-rod dystrophy beginning with night blindness followed by childhood-onset blindness, postaxial polydactyly, truncal obesity established during infancy and maintained through adulthood, renal abnormalities and learning difficulties, and a number of secondary features including anosmia and hepatic involvement. Cilia dysfunction leads to the loss of key signaling pathways essential for proper cellular function and has been shown to be directly associated with loss of vision, increased lipogenesis, and hyperphagia in this patient population. To date, no clear genotype-phenotype correlations have been identified (see Non-Patent Document 5). The current standard of care for BBS is management of clinical symptoms and supportive care for both patients and caregivers.

[0006] Targeting BBS through therapeutic modalities such as gene therapy and oligonucleotide therapy has been difficult due to the fact that many different genes may be mutated in BBS. Gene therapy efforts targeting the olfactory and retinal defects of BBS have had only modest success. In the ORPK mouse model, adenovirus-mediated expression of IFT88 restores cilia in the olfactory epithelium and improves olfactory responses (Non-Patent Document 6). A similar study in BBS1 mutant mice showed that AAV-mediated delivery of wild-type BBS1 restored cilia in olfactory sensory neurons and restored olfactory responses. However, the 60-90 day turnover of these sensory neurons, combined with the inability to administer multiple doses of AAV gene therapy vectors, limits the applicability of this approach. Furthermore, such intranasal delivery only reaches cells exposed at the apical surface of the olfactory epithelium and not deeper immature neurons (Non-Patent Document 7).

[0007] The genetic heterogeneity of BBS requires correcting each individual genetic defect, and as a result, dosage and toxicity must be established for each individual gene-specific therapeutic. Conversely, targeting the cilia defects underlying BBS regardless of genetic lesion site represents a treatment modality that can ameliorate multiple manifestations of BBS.

[0008] Sphingolipids and glycosphingolipids are key bioreactive molecules essential for the regulation of important cellular processes including differentiation, proliferation, aging, and cell-cell interactions. They are also central components of ciliary structure and contribute to ciliary signaling. Gangliosides GM1 and GM3 characterize distinct lipid microdomains in the apical membrane of epithelial cells, and ceramides are known to be abundant in the centrosomal / periocentriolar cell compartments. Ceramides also control the formation of primary cilia, and recent studies suggest that ciliary length may be controlled by the size or ceramide content of the ciliary base and its lipid flux to the cilium (Non-Patent Document 8).

[0009] The quinuclidine compounds described herein have activity as enzyme glucosylceramide synthase (GCS) inhibitors. Such compounds have utility in the treatment of conditions including lysosomal storage diseases, such as Gaucher's disease (e.g., US Pat. No. 5,399,623), proteinopathies, such as Alzheimer's disease (e.g., US Pat. No. 5,399,623), and cystic diseases, such as polycystic kidney disease (e.g., US Pat. No. 5,399,623). It has been suggested that quinuclidine compounds can act in these treatments, for example, by lowering glycolipid levels in the case of Gaucher's disease, or by reducing protein aggregation, for example, in the case of Alzheimer's disease, or by apoptosis, for example, in the case of polycystic kidney disease. The effect of these quinuclidine compounds on cilia, for example, on abnormal cilia associated with ciliopathies, has not been reported so far. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] WO2012 / 129084 [Patent Document 2] WO2016 / 145046 [Patent Document 3] WO2014 / 152215 [Non-Patent Document]

[0011] [Non-Patent Document 1] Marion, V et al., Proc. Natl. Acad. Sci. USA, 106(6):1820 - 26(2009) [Non-Patent Document 2] Aksanov, et al., Cell Mol. Life Sci., 71(17):3381 - 92(2014) [Non-Patent Document 3] Kulaga HM, et al., Nature Genetics, 36(9):944 - 48(2004) [Non-Patent Document 4] Tadenev AL et al., Proc. Natl. Acad. Sci. USA, 108(25):10320 - 25(2011) [Non-Patent Document 5] Haws R.et al, New Horizons in Translational Medicine, 2015, 2:102 - 109 [Non-Patent Document 6] McIntyre et al., Nature Med., 18(9):1423 - 28(2012) [Non-Patent Document 7] Williams CL et al., Molecular Therapy, 25(4):904 - 916(2017) [Non-Patent Document 8] Janich P.et al., FEBS Letters, 581(-):1783 - 1787(2007) [Summary of the Invention] [Problem to be solved by the invention]

[0012] There is a real need in the art to develop therapeutics that are effective in reducing or managing symptoms associated with ciliopathies, particularly ciliopathies such as BBS and Joubert Syndrome. There is also a particular need to develop therapeutics that are effective in treating the underlying pathophysiology of ciliopathies. [Means for solving the problem]

[0013] The present invention relates to a quinuclidine compound (Compound 1) according to formula (I) or a pharma- ceutically 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, where 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 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, where said alkyl or alkyloxy is halogen, hydroxy, cyano, and C 1~6 - optionally substituted by one or more (e.g., 1, 2 or 3) groups selected from alkyl, aryl, aryloxy ... A is halogen, hydroxy, thio, amino, nitro, C 1~6 Alkoxy or C 1~6 is a 5- or 6-membered aryl or heteroaryl group optionally substituted with 1, 2, or 3 groups independently selected from alkyl.

[0014] In a first aspect, the application provides a method for treating a ciliopathies in a subject in need thereof, comprising administering to the subject an effective amount of a quinuclidine compound as described herein, e.g., a compound according to Formula I. In a second aspect, the application provides a method for treating a disease or disorder selected from obesity, liver disease, retinal degeneration, olfactory dysfunction, hyperlipidemia, type 2 diabetes, and metabolic syndrome in a subject suffering from a ciliopathies, comprising administering to the subject an effective amount of a quinuclidine compound as described herein, e.g., a compound according to Formula I. In a third aspect, the application provides a method for preserving or improving ciliary function in a subject in need thereof, optionally a subject with a ciliopathies, comprising administering to the subject an effective amount of a quinuclidine compound as described herein, e.g., a compound according to Formula I.

[0015] Additional features and advantages of the compounds, compositions and methods disclosed herein will become apparent from the following detailed description. [Brief description of the drawings]

[0016] [Figure 1A]FIG. 1 shows quantification of cilia length in Wt and Bbs2− / − immortalized kidney epithelial cells. [Figure 1B] FIG. 1 shows immunofluorescence analysis of GSL localization in Wt and Bbs2− / − immortalized kidney epithelial cells. [Figure 1C] FIG. 1 shows the effect of treatment with Compound 1 on cilium length and levels of GM3 cilium in Wt and Bbs2− / − immortalized kidney epithelial cells. [Diagram 2] FIG. 1 shows the effect on glucosylceramide (GlcCer or GL1) levels in multiple different tissues in Wt and Bbs2− / − mouse models when treated with Compound 1. [Figure 3A] FIG. 1 shows changes in metabolic parameters including food consumption, body weight, body fat percentage and serum leptin measured in Wt mice, Bbs2− / − mice and Bbs2− / − mice treated with compound 1 from 1 to 6 months of age. [Figure 3B] Representative H&E stained white adipose tissue (top) and quantification of adipocyte volume (bottom) from Wt mice, Bbs2- / - mice, and Bbs2- / - mice treated with compound 1 at 1 to 6 months of age. [Figure 3C] FIG. 1 shows mRNA analysis of pro-adipogenic genes in white adipose tissue from 1- to 6-month-old Wt, Bbs2− / −, and Bbs2− / − mice treated with Compound 1. [Figure 4A] FIG. 1 shows the effect of short-term treatment with Compound 1 on metabolic parameters including body weight, percentage of body fat and serum leptin measured in Bbs2− / − mice (4-6 month old mice) exhibiting established metabolic disease. [Figure 4B]FIG. 1 shows representative H&E stained white adipose tissue and quantification of adipocyte volume from Wt mice, Bbs2− / − mice with established metabolic disease (mice aged 4 to 6 months), and Bbs2− / − mice with established metabolic disease (mice aged 4 to 6 months) treated with short-term treatment with Compound 1. [Diagram 5] FIG. 1 shows analysis of cilia in the hypothalamus of Wt, Bbs2− / − and Bbs2− / − mice treated with Compound 1 in the diet. [Figure 6] FIG. 1 shows changes in liver parameters including liver weight, serum ALT and serum triglycerides measured in Wt mice, Bbs2− / − mice and Bbs2− / − mice treated with compound 1 from 1 month to 6 months of age. [Figure 7A] FIG. 13 shows optical coherence tomography analysis of outer nuclear layer (ONL) thickness in Wt and Bbs2− / − mice. [Figure 7B] FIG. 1 shows an analysis of the outer nuclear layer (ONL) / inner nuclear layer (INL) ratio in Wt mice, Bbs2− / − mice, and Bbs2− / − mice treated with Compound 1 at 1 to 6 months of age. [Figure 7C] FIG. 1 shows analysis of the expression of rhodopsin (top) and cone arrestin (bottom)—specific for rods and cones, respectively—in eye sections from 1- to 6-month-old Wt, Bbs2− / −, and Bbs2− / − mice treated with compound 1. [Figure 8A] FIG. 1 shows the results of an in vivo buried treat test used to measure the latency to remove a hidden treat in 1- to 6-month-old Wt, Bbs2− / −, and Bbs2− / − mice treated with Compound 1. [Figure 8B]FIG. 1 shows analysis of acetylated tubulin, a marker of cilia, in nasal sections from 1- to 6-month-old Wt, Bbs2− / −, and Bbs2− / − mice treated with Compound 1. [Figure 8C] FIG. 1 shows analysis of adenylate cyclase III, a marker of odorant signaling, in nasal sections from 1- to 6-month-old Wt, Bbs2− / −, and Bbs2− / − mice treated with compound 1. [Figure 9] FIG. 1 shows analysis of markers of cell layers of the main olfactory epithelium (MOE) in nasal sections from 1- to 6-month-old Wt, Bbs2− / − and Bbs2− / − mice treated with compound 1, namely horizontal basal cells (CK14, cytokeratin 14), globular basal and supporting cells (Sox2, SRY-Box2), immature neurons (Dcx, doublecortin) and mature neurons (OMP, olfactory marker protein). [Figure 10A] FIG. 1 shows an in vitro assay of human adipocyte differentiation showing accumulation of lipid (globular) in mature adipocytes upon knockdown of BBS1, BBS2, and BBS10 genes using siRNA. [Figure 10B] FIG. 13 shows an in vitro assay of human adipocyte differentiation showing that leptin concentration in adipocyte conditioned medium is higher after knockdown of the BBS gene. [Figure 10C] FIG. 1 shows an in vitro assay of human adipocyte differentiation showing a dose-dependent effect on lipid (globular) accumulation by treatment with compound 1 (1.25-10 μM). [Figure 10D] FIG. 1 shows an in vitro assay of human adipocyte differentiation showing a dose-dependent effect on leptin secretion by treatment with compound 1 (1.25-10 μM). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Specific embodiments of the present disclosure will now be described with reference to preparations and schemes, but it should be understood that such embodiments are merely illustrative and are merely a few of the many possible specific embodiments that can represent applications of the principles of the present disclosure by way of example. Various changes and modifications will be apparent to those skilled in the art upon consideration of 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.

[0018] 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 may be used to carry out or test the present invention, exemplary methods, devices, and materials are described herein. 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.

[0019] 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.

[0020] All numerical indications, including ranges, such as pH, temperature, time, concentration, molecular weight, are approximate values ​​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 indications are preceded by the term "about". It is also understood, although not always explicitly stated, that the reagents described herein are merely exemplary, and that equivalents of such are known in the art.

[0021] As used herein, the term "optionally substituted" is meant to be equivalent to the phrase "unsubstituted or substituted with."

[0022] 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").

[0023] 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 herein to mean the phrase "including, but not limited to," and is used interchangeably.

[0024] As used herein, the terms "comprising" or "comprising" are intended to mean that the compositions and methods include the recited elements but do not exclude others. "Consisting essentially of" is intended to mean, when defining compositions and methods, excluding any other elements that are essentially important to the combination for the purposes described. Thus, a composition that consists essentially of the elements defined herein will not exclude trace contaminants from the isolation and purification methods, as well as pharma- ceutically 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 method steps for producing the compositions or achieving the intended results. Embodiments defined by each of these transition terms are within the scope of the present invention. The use of the term "comprising" herein is intended to encompass "consisting essentially of" and "consisting of."

[0025] The term "ciliopathies" refers to diseases characterized by ciliary dysfunction. "Ciliary dysfunction" refers to abnormal formation and / or function of cilia, including abnormalities in ciliary position. Ciliary dysfunction may affect the extracellular and / or intracellular portions of the cilia, which may be characterized by structural and / or functional irregularities.

[0026] "Subject", "individual" or "patient" are used interchangeably herein to refer to vertebrates, such as 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 one embodiment, the mammal is a human.

[0027] "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 mucosa), by injection (e.g. subcutaneous, intravenous, parenteral, intraperitoneal, CNS), or by inhalation (e.g. oral or nasal). Pharmaceutical preparations are of course provided in a form suitable for each administration route.

[0028] "Treating" a disease or "treatment" thereof includes (1) inhibiting the disease, i.e., arresting or reducing the onset of the disease or its clinical symptoms; and / or (2) relieving the disease, i.e., causing regression of the disease or its clinical symptoms.

[0029] "Preventing" a disease or "prevention" includes preventing the development of clinical symptoms of the disease in patients who may be susceptible to the disease but who have not yet experienced or exhibited symptoms of the disease.

[0030] The term "suffering" in conjunction with the term "treatment" refers to a patient or individual who has been diagnosed with a disease. The term "suffering" in conjunction with the term "prevention" refers to a patient or individual who is susceptible to a disease. A patient may also refer to one who is "at risk for" a disease because of a history of the disease in their family or because of the presence of a genetic mutation associated with the disease. A patient at risk for a disease has not yet developed all or some of the characteristic symptoms of the disease.

[0031] An "effective amount" or a "therapeutically effective amount" is an amount sufficient to produce a beneficial or desired result. 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 the individual dosage units, the bioavailability of the therapeutic agent, and the route of administration. However, it is understood that the specific dosage level of the therapeutic agent of the present invention for any particular subject will depend on a variety of factors, including, for example, the activity of the specific compound used, the subject's age, weight, general health, sex, and diet, the time of administration, the rate of excretion, the drug combination, and the severity and form of administration 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 the appropriate dose to administer to a patient. In general, it will be desired to administer an effective amount of the compound to achieve serum levels commensurate with the concentration 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 ciliopathies, ex vivo, in vitro or in vivo.

[0032] As used herein, the term "pharmaceutical 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, and 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 Pharmaceutical Sciences (20th ed., Mack Publishing Co. 2000).

[0033] 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 an active drug. For example, a prodrug is a variant or derivative of a quinuclidine compound described herein that has a group that can be cleaved under certain metabolic conditions, which, when cleaved, becomes a quinuclidine compound described herein, such as a compound of formula I. Such prodrugs are then pharmacologic in vivo active 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 the advantages of solubility, tissue compatibility, or delayed release in mammalian organisms.

[0034] Prodrugs generally known in the art include well-known acid derivatives such as, for example, esters prepared by reaction of an acid compound with a suitable alcohol, and amides prepared by reaction of an acid compound with an amine, which is a basic group that reacts to form an acylated base derivative. 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 that have an amino acid residue or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues that are covalently linked to the free amino, hydroxy, or carboxylic acid groups of the compounds disclosed herein via peptide bonds. Amino acid residues include the 20 naturally occurring amino acids, commonly designated by their three letter symbols, and also include 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.

[0035] As used herein, the term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable acid addition salts or pharmaceutically acceptable base addition salts of the presently disclosed compounds which may be administered without any substantial undesirable biological effects or any adverse interactions as a result with other components of the pharmaceutical composition in which it may be included.

[0036] As used herein, "C 1~6 The term "-alkyl" refers to a saturated linear or branched free radical consisting essentially of one to six carbon atoms and a corresponding number of hydrogen atoms.1-6 -Alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, and isobutyl. Other C 1~6 -alkyl groups will be readily apparent to those of 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 one to three (or four) carbon atoms and the corresponding number of hydrogen atoms.

[0037] 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.

[0038] 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.

[0039] As used herein, "C 1~6The term "-alkyloxy" refers to 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 The -alkyloxy group is bonded via an oxygen atom. 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 skill in the art given the benefit of this disclosure. 1~3 -alkyloxy", "C 1~4 The terms "-alkyloxy" and the like have the equivalent 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 bonded via an oxygen atom.

[0040] 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 a 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 -alkenyloxy group is ethenyloxy; others will be readily apparent to the skilled artisan given the benefit of this disclosure.

[0041] 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 a 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-alkenyloxy group is ethynyloxy; others will be readily apparent to the skilled artisan given the benefit of this disclosure.

[0042] As used herein, the term "heteroaryl" refers to an aromatic free radical having 5 or 6 atoms (i.e., ring atoms) forming a ring, with 1 to 5 ring atoms being carbon, and the remaining 1 to 5 ring atoms (i.e., hetero ring atoms) being 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 one 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, it will be understood by those skilled in the art that certain other atoms, such as heterocyclic atoms, can be attached to the main structure.

[0043] 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.

[0044] As used herein, the term "aliphatic" refers to a non-aromatic compound containing carbon and hydrogen atoms, e.g., 1 to 9 carbon atoms. An aliphatic compound 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 having aromatic character). Examples of aliphatic compounds include ethane, propylene, cyclobutane, and cyclohexadiene.

[0045] As used herein, the terms "halo" and "halogen" refer to fluorine, chlorine, bromine, or iodine. These terms are used interchangeably and may refer to halogen free radical groups or halogen atoms themselves. Those skilled in the art will be able to easily confirm the identity in light of the context in which the terms are used in this disclosure.

[0046] 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.

[0047] As used herein, the term "nitro" refers to the -NO2 radical attached through the nitrogen atom.

[0048] 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.

[0049] As used herein, the term "amino" refers to a free radical having a nitrogen atom and one or two hydrogen atoms. Thus, the term "amino" generally refers to primary and secondary amines. In that respect, as used herein, tertiary amines are represented by the general formula RR'N-, where 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 the term is used in this disclosure.

[0050] As used herein, the term "oxo" refers to an oxygen radical linked through a double bond. If 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.

[0051] The recitation of a list of chemical groups in any definition of a variable herein includes definitions of the variable as any single group or combination of listed groups. The recitation of an embodiment of a variable or aspect herein includes that embodiment in any single embodiment or in combination with any other embodiment or portion thereof.

[0052] Any composition or method provided herein may be combined with any one or more of the other compositions and methods provided herein.

[0053] The following abbreviations are used herein: br Broad signal CDI Carbonyldiimidazole CNS Central Nervous System d doublet DAPI 4',6-diamidino-2-phenylindole dd Doublet of Doublets DME Dimethoxyethane DMEM Dulbecco's Modified Eagle's Medium DMSO-d6 Dimethylsulfoxide-d6 DMF Dimethylformamide DNA deoxyribonucleic acid DTBZ Carbon-11 dihydrotetrabenazine EDTA Ethylenediaminetetraacetic acid ELISA Enzyme-linked Immunosorbent Assay Et2O Diethyl ether EtMgBr Ethyl magnesium bromide EtOAc Ethyl acetate GL1 Glucosylceramide (GlcCer) GM1 monosialotetrahexosylganglioside GM3 monosialylated dihexosylganglioside GSL Glycosphingolipids H&E Hematoxylin and Eosin Staining HPLC High Pressure / Performance Liquid Chromatography HAS 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 TWEEN20 Polysorbate 20 TWEEN80 Polysorbate 80 Wt wild type UPLCMS Ultra-Performance Liquid Chromatography Mass Spectrometry

[0054] compound The present invention relates to quinuclidine compounds for use in the treatment of ciliopathies. In all its various aspects, the present invention relates to a quinuclidine compound (Compound 1) according to formula (I) or a pharma- ceutically 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, where 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 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, where said alkyl or alkyloxy is halogen, hydroxy, cyano, and C 1~6- optionally substituted by one or more (e.g., 1, 2 or 3) groups selected from alkyl, aryl, aryloxy ... A is halogen, hydroxy, thio, amino, nitro, C 1~6 Alkoxy and C 1~6 and alkyl, aryl, aryl group ...

[0055] In further embodiments of any of the aspects of the invention, 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, where 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, where 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, C 1~4 -alkyloxy, where 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; 1.4 Compound 1 (wherein R 1 is hydrogen, halogen, C 1~4 -Alkyl, C 1~4-alkyloxy, where 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 or amino; 1.5 Compound 1 (wherein R 1 is hydrogen, halogens, and C 1~4 -alkyl, wherein said alkyl is optionally substituted with one or more (e.g., 1 or 2) groups selected from halogen, hydroxy, thio, or 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 or 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 each independently represents a C optionally substituted with one or more (e.g., 1, 2, or 3) halogens; 1~3 - alkyl); 1.11 Compound 1.11 (wherein R 2 and R 3 are each independently methyl or ethyl optionally substituted with 1 or 2 halogens; 1.12 Compound 1.11 (wherein R 2 and R 3are each independently selected from methyl and ethyl optionally substituted with one or more fluorines, e.g., 1, 2 or 4 fluorines; 1.13 Compound 1.11 (wherein R 2 and R 3 are each independently methyl substituted with 0, 1, 2 or 3 fluorines; 1.14 Compound 1.11 (wherein R 2 and R 3 are each methyl or trifluoromethyl); 1.15 Compound 1.11(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., 1 or 2) 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 R 3 together form a cyclopropyl group); 1.19 Compound 1 or any of 1.1 to 1.9 (wherein R 4 , R 5 and R 6 is hydrogen, halogen, C 1~6 -Alkyl, and C 1~6 -alkyloxy, where said alkyl or alkyloxy is 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, where said alkyl or alkyloxy is 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, where 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 -alkyloxy) 1.24 Compound 1 or any of 1.19 to 1.23 (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 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 1.26(R 4 is 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(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); 1.28 Compound 1.26 (wherein R 4 is fluorine or one or more (e.g., 1, 2 or 3) C 1~3 -ethoxy optionally substituted with 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 hydrogen); 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 with alkyloxy (e.g. methoxy); 1.33 Compound 1.32 (wherein R 5 and R 6 are hydrogen, and R 4 is ethoxy substituted with fluorine, or 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, wherein 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 to 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 (e.g., ortho, meta, or para to the A substituent) of the phenyl ring to which it is attached); 1.40 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- 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 None of R is hydrogen. 4 , R 5 and R 6 are independently located at the 2-, 4-, or 6-position (i.e., ortho or para to the A substituent) of the phenyl ring to which they are attached); 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, where A is a 6-membered aryl group, a 5-membered heteroaryl group (e.g., containing 1, 2 or 3 heteroatoms in the heteroaryl ring 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 selected from N, O and S), optionally the 5-membered heteroaryl group containing 1 or 2 heteroatoms selected from N and S (e.g., one N and / or one S); 1.46 The 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, where 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, where A is a halogen (e.g., fluorine) and C 1~6 phenyl substituted with 1, 2, or 3 groups independently selected from alkyl (e.g., methyl); 1.53 Compound 1.52, where A is phenyl substituted with one or two fluorines or methyl groups; 1.54 Compound 1 or any of 1.1-1.53 ​​(wherein the two groups bonded 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 substituent 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 to 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 at least one of the aryl- and aryl-groups) is bonded to a carbon atom of the heteroaryl ring, and optionally both such groups are bonded to a carbon atom of the heteroaryl ring;

[0056] 1.58 Compound 1, or any of 1.1-1.57, wherein the compound of formula I may be represented by any one or more of the following structures: [ka] [ka] [ka]

[0057] 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 1.1-1.58, 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%, 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 racemic (i.e., approximately a 50:50 ratio of enantiomers) or is some other ratio (e.g., less than 50:50 or more than 50:50) of a mixture of enantiomers;

[0058] 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]

[0059] 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 1.1-1.63, 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 1.1-1.63, wherein the compound is 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 compound I, or any of II to XII, is in the form of the free base; 1.68 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 pharma- ceutically 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)thiazol-4-yl)propan-2-yl)carbamate in the malate form; 1.74 Compound 1, or any of 1.1-1.73, wherein the compound of formula I, or any of II-XII, is in the form of a prodrug as described herein; 1.75 Compound 1, or any of 1.1-1.74, wherein the compound of formula I, or any of II to XII, is in the form of a hydrate, solvate and / or polymorph.

[0060] salt The compounds disclosed herein, e.g., any of compounds 1 or 1.1 to 1.75, are basic in nature and can generally form a variety of different salts with various inorganic and / or organic acids. Such salts are generally pharma- ceutically acceptable for administration to animals and humans, but it is often actually desirable to first isolate the compound from the reaction mixture as a pharma- ceutically unacceptable salt, then simply convert the latter back to a free base compound by treatment with an alkaline reagent, and then convert the free base to a pharma-ceutically acceptable acid addition salt. Acid addition salts of basic compounds may be readily prepared using conventional techniques, e.g., 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, e.g., methanol or ethanol. When the solvent is carefully evaporated, the desired solid salt is obtained. The compounds disclosed herein are positively charged, and include, e.g., quaternary ammonium, and can also form salts with anionic components of various inorganic and / or organic acids.

[0061] Acids which may be used to prepare pharma- ceutically acceptable salts of the 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.

[0062] Compounds disclosed herein that are acidic in nature, e.g., compounds that contain a thiol moiety, can generally form a variety of different salts with various inorganic and / or organic bases. Although such salts are generally pharma-ceutically acceptable for administration to animals and humans, it is often actually desirable to first isolate the compound from the reaction mixture as a pharma-ceutically 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 pharma-ceutically acceptable base addition salt. These base addition salts may be readily prepared using conventional techniques, e.g., 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 and the desired alkali metal alkoxide together, 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.

[0063] Bases which may be used to prepare pharma- ceutically acceptable base addition salts of the 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, such as N-methylglucamine (meglumine), lower alkanolammonium, and other organic amine bases.

[0064] In one embodiment, the pharmaceutically acceptable salt is a succinate salt. In another embodiment, the pharmaceutically acceptable salt is a 2-hydroxysuccinate salt, such as (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.

[0065] Prodrug The present disclosure further includes prodrugs of compounds 1 and 1.1 to 1.75. The pharma- ceutically acceptable prodrugs disclosed herein are derivatives of quinuclidine compounds that can be converted in vivo to the quinuclidine compounds described herein. Prodrugs may have some activity themselves, for example, when they undergo solvolysis under physiological conditions or enzymatic degradation, they become pharma- ceutical active in vivo. Methods for preparing prodrugs of the compounds described herein will be clear to those skilled in the art based on the present disclosure.

[0066] 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 may 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, for example 1,2-ethanediol.

[0067] 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 the 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.

[0068] 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.

[0069] In one embodiment, the quinuclidine-3-yl group of the quinuclidine compounds defined herein has the R-configuration. Thus, the quinuclidine compounds may be selected from the group consisting of compounds of formulae (Ia) to (XIIa), and pharma-ceutically acceptable salts and prodrugs thereof:

[0070] [ka] [ka] [ka]

[0071] In another embodiment, the quinuclidine-3-yl group of the quinuclidine compounds defined herein has the S-configuration. Thus, the quinuclidine compounds may be selected from the group consisting of compounds of formulae (Ib) to (XIIb), and pharma-ceutically acceptable salts and prodrugs thereof:

[0072] [ka] [ka] [ka]

[0073] In one embodiment, the quinuclidine compound is a compound of formula (Xb) or a pharma- ceutically acceptable salt or prodrug thereof. In another embodiment, the quinuclidine compound is a compound of formula (XIIb) or a pharma- ceutically acceptable salt or prodrug thereof.

[0074] In one embodiment, the quinuclidine-3-yl group of the quinuclidine compound defined herein exists in 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 the 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 pharma-ceutically 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 exist in approximately equal amounts. In another embodiment, the quinuclidine compound exists as a mixture of isomers with R- and S-configuration, and R- and S-isomers exist in different amounts.In one embodiment, the S-isomer exists 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 exists in an enantiomeric excess of at least about 5%, 10%, 25%, 40%, 70%, 80%, 90%, 95%, 97%, 98% or 99%, for example, about 100%.

[0075] Methods for preparing enantiomerically enriched and / or enantiopure quinuclidine compounds will be apparent to those of skill in the art based on the present disclosure.

[0076] The compounds disclosed herein may exist in several tautomers, including enol and imine forms, and keto and enamine forms, as well as geometric isomers, and mixtures thereof. Tautomers exist as a mixture of tautomer 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.

[0077] Atropisomers are also within the scope of this disclosure. Atropisomers refer to compounds that can be decomposed into isomers with restricted rotation.

[0078] 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 quinuclidine compounds described herein may be in amorphous form and / or in one or more crystalline forms.

[0079] Isotopically labeled compounds are also within the scope of the present disclosure.As used herein, "isotopically labeled compounds" refers to compounds disclosed herein, including pharmaceutical salts and their prodrugs, each of which is described herein, in which 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.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.

[0080] Medical Indication The quinuclidine compounds described herein and pharmaceutical compositions comprising them are useful in therapy, particularly in the therapeutic treatment of ciliopathies in subjects.The subjects that will be treated according to the methods described herein include vertebrates, such as mammals.In certain embodiments, the mammal is a human patient.

[0081] In a first aspect, the invention provides a method for treating a ciliopathies in a subject in need thereof (Method 1), comprising administering to the subject an effective amount of a quinuclidine compound as described herein, e.g., 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. Also provided is a quinuclidine compound as described herein, e.g., a compound according to formula I or any of II-XIIIa-XIIa or Ib-XIIb, or any of compounds 1 or 1.1 to 1.75, for use in the method of treating a ciliopathies in a subject, e.g., for use in Method 1 or 1.1 to 1.62. Further provided is the use of a quinuclidine compound as 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 a ciliopathic disorder in a subject, e.g., in the manufacture of a medicament for use in any of Methods 1 or 1.1-1.62.

[0082] 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 Method 1, comprising administering to a subject an effective amount of Compound 1 or any one or more of Compounds 1.1 to 1.75; 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 1.1-1.75; 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 1.1-1.75; 1.5 Method 1.3 or 1.4, wherein the pharmaceutical composition further comprises at least one pharma- ceutically acceptable excipient as described herein; 1.6 Any of Methods 1 or 1.1-1.5, wherein the method comprises administering an effective amount of a compound or a pharmaceutical dosage form comprising an effective amount of a pharmaceutical composition; 1.7 Method 1.6, where 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, in which the dosage form is a chewable tablet; 1.9 Method 1.6, where the dosage form is a parenteral dosage form (e.g., the pharmaceutical composition is formulated for injection); 1.10 The injection is intravenous, intramuscular, intrathecal or subcutaneous, optionally a sterile injection, Method 1.9; 1.11 Method 1.6, where 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 simultaneously administering in a patient in need thereof a second active agent as described herein, e.g., a second compound capable of treating or preventing a ciliopathies; 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 Method 1, or any of 1.1-1.14, wherein the subject is a mammal; 1.16 The subject is a primate, method 1.15; 1.17 The subject is a human, method 1.16; 1.18 Any of methods 1 or 1.1 to 1.17, wherein the ciliopathy is a disease selected from the group consisting of Joubert syndrome, Meckel-Gruber syndrome, Senior-Loken syndrome, orofacial-digital syndrome type I, Leber congenital amaurosis, Bardet-Biedl syndrome (BBS), Amström syndrome, June asphyxiating thoracic dystrophy, Ellis-van Creveld syndrome, Sensenbrenner syndrome, and primary ciliary dyskinesia, or a combination thereof; 1.19 The ciliary disease is BBS, either method 1 or 1.1-1.18; 1.20 The ciliopathy is Meckel-Gruber syndrome, method 1.18 or 1.19; 1.21 The ciliopathy is Senior-Loken syndrome, any of methods 1.18-1.20; 1.22 The ciliopathies are Joubert syndrome, any of methods 1.18-1.21; 1.23 The ciliopathy is Leber congenital amaurosis, any of methods 1.18 to 1.22; 1.24 The ciliopathy is orofacial-digital syndrome type I, any of methods 1.18-1.23; 1.25 The ciliopathies is Amström syndrome, any of methods 1.18-1.24; 1.26 The ciliopathy is Ellis-van Creveld syndrome, any of methods 1.18-1.25; 1.27 The ciliopathy is Sensenbrenner syndrome, any of methods 1.18-1.26; 1.28 The ciliopathies is primary ciliary dyskinesia, any of methods 1.18 to 1.27; 1.29 Any of methods 1 or 1.1-1.28, wherein the subject has been diagnosed with a mutation in one or more of the genes BBS1 (ARL), BBS2, BBS3, BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9 (B1), BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (C2ORF86 / FRITZ), BBS16 (SDCCAG8), BBS17, BBS18, BBS19, BBS20, and BBS21; 1.30 Method 1.29, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1, BBS2, BBS4, BBS5, BBS7, BBS8, BBS9, BBS10, and BBS18; 1.31 Method 1.30, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1, BBS2, and BBS10; 1.32 Method 1.31, wherein the subject has been diagnosed with a mutation in at least gene BBS2; 1.33 Any of methods 1 or 1.1-1.32, wherein the subject has been diagnosed with a mutation in one or more of the genes MKS1, MKS3, CEP290, RPGRIP1L, CC2D2A, and TMEM216; 1.34 Any of methods 1 or 1.1 to 1.32, wherein the subject has been diagnosed with a mutation in at least the gene MKS1; 1.35 Any of methods 1 or 1.1-1.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, TTC21B, KIF7, TCTN1, TMEM237, CEP41, TMEM138, C5ORF42, TCTN3, ZNF423, TMEM231, CSPP1, ARMC9, INPP5E, CXORF5, INVS, NPHP3, NPHP4, NPHP5(IQCB1), and SDCCAG8; 1.36 Any of methods 1 or 1.1-1.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, TTC21B, KIF7, TCTN1, TMEM237, CEP41, TMEM138, C5ORF42, TCTN3, ZNF423, TMEM231, CSPP1, ARMC9, INPP5E, and CXORF5; 1.37 Any of methods 1 or 1.1-1.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, INPP5E, and CXORF5; 1.38 Method 1 or any of 1.1 to 1.37, wherein the subject has been diagnosed with a mutation in one or more of the genes AHI1, ARL13B, INPP5E, and OFD1. 1.39 Any of methods 1 or 1.1-1.38, wherein the subject has been diagnosed with a mutation in one or more of the genes CEP290, NPHP1, INVS, NPHP3, NPHP4 and NPHP5. 1.40 The subject has been diagnosed with a mutation in gene OFD1, any of methods 1 or 1.1 to 1.39; 1.41 The subject has been diagnosed with a mutation in one or more of the genes GUCY2D, RPE65, SPATA7, AIPL1, LCA5, RPGRIPL1, CRX, CRB1, IMPD1, RD3, CEP290, NPHP5, and RDH12, according to any of methods 1 or 1.1-1.40; 1.42 Any of methods 1 or 1.1-1.40, wherein the subject has been diagnosed with a mutation in one or more of the genes GUCY2D, RPE65, SPATA7, AIPL1, LCA5, CRX, CRB1, IMPD1, RD3, and RDH12; 1.43 The subject has been diagnosed with a mutation in the gene ALMS1, according to any of methods 1 or 1.1 to 1.42; 1.44 The subject has been diagnosed with a mutation in gene IFT80, according to any of methods 1 or 1.1 to 1.43; 1.45 Any of methods 1 or 1.1-1.44, wherein the subject has been diagnosed with a mutation in one or more of the genes EVC1, EVC2, IFT122, IFT43 and WDR35; 1.46 Method 1.45, wherein the subject has been diagnosed with a mutation in one or more of the genes IFT122, IFT43 and WDR35; 1.47 Method 1.33, wherein the subject has been diagnosed with a mutation in one or more of genes EVC1 and EVC2; 1.48. Any of methods 1 or 1.1-1.47, wherein the subject has been diagnosed with a mutation in one or more of the genes DNAI1, DNAH5, TXNDC3, DNAH11, DNAI2, KTU, RSPH4A, RSPH9, and LRRC50; 1.49 Method 1, or any of 1.1-1.48, wherein the subject suffers from a comorbidity selected from obesity, liver disease, retinal degeneration, olfactory deficit, hyperlipidemia, type 2 diabetes, and metabolic syndrome; 1.50 The subject also suffers from a glycolipid storage or accumulation disease or disorder, any of methods 1, or 1.1-1.49; 1.51 The glycolipid storage or accumulation disease or disorder is selected from polycystic kidney disease (PKD) (e.g., autosomal dominant PKD [ADPKD]), gangliosidosis (e.g., GM1 gangliosidosis or GM2 gangliosidosis or GM3 gangliosidosis), Gaucher disease (e.g., Gaucher type 1, Gaucher type 2, or Gaucher type 3), Fabry disease, and Parkinson's disease (e.g., Duchenne Parkinson's disease), method 1.50; 1.52 The subject is also being treated with an enzyme replacement therapy (ERT), e.g., using glucocerebrosidase (e.g., alglucerase, imiglucerase, velaglucerase, or taliglucerase), alpha-galactosidase (e.g., agalsidase alpha or agalsidase beta), or beta-galactosidase, each of which optionally is a recombinant enzyme, Method 1, or any of 1.1-1.50; 1.53 Method 1, or any of 1.1-1.52, wherein the subject is administered a daily dose of a compound 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; 1.54 Method 1, or any of 1.1-1.53, 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; 1.55 any of Methods 1, or 1.1-1.54, wherein the method is effective to treat, reduce, or ameliorate one or more symptoms or signs selected from obesity, liver disease (e.g., elevated serum liver enzymes, e.g., ALT, AST, alkaline phosphatase, gamma glutamyl transpeptidase), retinal degeneration, hyperlipidemia (e.g., elevated serum total cholesterol, LDL, VLDL, or triglycerides), type 2 diabetes (e.g., elevated serum glucose), and olfactory dysfunction; 1.56 Any of Methods 1, or 1.1.-1.55, wherein the method is effective to preserve or improve ciliary function in the hypothalamus, retina and / or olfactory epithelium, e.g., to preserve or improve cilia function (e.g., motility) and / or to preserve or improve the amount or density of functioning cilia; 1.57 Method 1, or any of 1.1-1.56, wherein the compound, or a pharma- ceutically 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 Any of Methods 1, or 1.1-1.56, wherein the compound, or a pharma- ceutically acceptable salt or prodrug thereof, is administered by localized administration, e.g., by topical administration; 1.61 Method 1, or any of 1.1-1.60, wherein the compound is (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate; 1.62 Method 1, or any of 1.1-1.61, 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.

[0083] Diseases and disorders, such as ciliopathies, are often associated with one or more genetic mutations. In some embodiments of the present disclosure, a subject, i.e., a patient, is diagnosed with a particular disease or disorder, and is also diagnosed with a particular genetic mutation, e.g., one that is known to cause the disease or disorder in question, but it is often not possible to prove that a particular patient's disease or disorder is caused by the particular mutation that the person has been diagnosed with. When used in this way, the term "diagnosed with a particular genetic mutation" means that the subject, i.e., patient, has been tested, e.g., by DNA or RNA sequencing, protein profiling, or other suitable means, and is found to have the mutation in question. However, as further described below, many genetic diseases and disorders may have multiple genetic causes (e.g., mutations), and a patient may have multiple mutations that may be sufficient to cause the disease or disorder, respectively, under some circumstances, and is not subject to confirm that a particular mutation causes a particular disease or disorder in a particular patient.

[0084] Bardet-Biedl syndrome and Meckel-Gruber syndrome Bardet-Biedl syndrome (BBS) is a rare autosomal recessive multisystem genetic disorder (see Waters et al., Pediatr. Nephrol., 2011, 26:1039-1056). The prevalence of BBS in the United States and Northern Europe is 1:160,000. The main features of BBS include rod-cone dystrophy, polydactyly, obesity, learning disability, hypogonadism, and renal abnormalities. BBS can result from mutations in at least 21 different genes, although mutations in BBS1, BBS2, and BBS10 account for approximately 50% of cases. The genes affected in BBS are essential for the assembly of the BBSome, a macromolecular complex that is a component of the basal body and is involved in the formation, maintenance, and function of primary cilia.

[0085] Meckel-Gruber syndrome is an autosomal recessive lethal malformation that phenotypically overlaps with other ciliary diseases (Waters et al., supra). Clinical features include occipital encephalocele and other posterior fossa defects, cystic dysplastic kidneys, hepatobiliary proliferation, and polydactyly. Meckel-Gruber syndrome is caused by mutations in several genes, including MKS1, MKS3, CEP290, RPGRIP1L, CC2D2A, and TMEM216. Mutations in MKS1 have also been implicated in BBS.

[0086] Thus, in an embodiment, the ciliopathies are selected from BBS and Meckel-Gruber syndrome. In one embodiment, the ciliopathies are BBS. In another embodiment, the ciliopathies are Meckel-Gruber syndrome.

[0087] Joubert syndrome and Senior-Loken syndrome Joubert syndrome is a rare autosomal recessive genetic disorder affecting the cerebellum. It is characterized by hypotonia, ataxia, psychomotor retardation, irregular breathing patterns and oculomotor apraxia. Joubert syndrome shares phenotypic and genotypic overlap with Senior-Loken syndrome, a rare autosomal recessive disorder characterized by nephrophthisis and progressive eye disease (Waters et al., supra).

[0088] Thus, in an embodiment, the ciliopathies are selected from Joubert syndrome and Senior-Loken syndrome. In one embodiment, the ciliopathies are Joubert syndrome. In another embodiment, the ciliopathies are Senior-Loken syndrome.

[0089] Orofacial-digital syndrome type I Orofacial-digital syndrome type 1, also known as Papillon League and Psomme syndrome, is a rare X-linked congenital disorder. Mutations in the OFD1 gene have been described in patients with orofacial-digital syndrome type 1. OFD1 encodes a centrosomal protein that localizes to the basal body at the origin of the primary cilium, and OFD1 localizes to both centrosomes and basal bodies in human genetic cytoarchitectures. Reduced ciliogenesis has been observed with disease-associated mutations (Waters et al., supra).

[0090] Leber congenital amaurosis Leber congenital amaurosis is a severe retinal dystrophy that occurs in the first year of life. Visual function is often poor and is frequently associated with nystagmus, reduced or near-absent pupillary response, photophobia, hyperopia, and keratoconus (Waters et al., supra).

[0091] Amström syndrome Amström syndrome is a rare autosomal recessive disorder characterized by multiple organ dysfunction including cone-rod dystrophy, obesity, progressive sensorineural hearing loss and dilated cardiomyopathy. Amström syndrome is caused by mutations in the gene ALMS1, which encodes a protein that is specifically localized to the proximal ends of centrioles and basal bodies (Waters et al., supra). The ALMS1 protein is involved in ciliary function, cell cycle control and intracellular trafficking.

[0092] June Asphyxiating Thoracic Dystrophy June asphyxiating thoracic dystrophy (June syndrome) is a rare autosomal recessive chondrodysplasia that affects the development of cartilage and bone in children. June syndrome can be caused by mutations in IFT80, which has been shown to localize to the basal bodies of cilia in a murine chondrocyte cell line (Waters et al., supra).

[0093] Ellis-van Creveld syndrome and Sensenbrenner syndrome Ellis-van Creveld syndrome is a rare chondroectodermal dysplasia characterized by skeletal abnormalities including postaxial polydactyly, short ribs, cleft palate, and carpal malformations. It can be caused by mutations in EVC1 or EVC2. EVC proteins have been shown to localize to the base of the primary cilium in chondrocytes (Waters et al., supra). Sensenbrenner syndrome (also known as cranioectodermal dysplasia) is an autosomal recessive disorder that is similar to Ellis-van Creveld syndrome. It can be caused by mutations in IFT122, IFT43, or WDR35, all of which code for ciliary proteins (Waters et al., supra).

[0094] Primary ciliary dyskinesia Primary ciliary dyskinesia (also known as Kartagener syndrome) is a rare autosomal recessive disorder that results in defects in the action of cilia lining the airways (lower and upper, paranasal sinuses, Eustachian tubes, middle ear), fallopian tubes, and the flagella of sperm cells.

[0095] The method of the present invention may be beneficial for subjects who have been diagnosed with ciliopathies but have not yet experienced the typical symptoms associated with pathology.The method of the present invention may also be beneficial for subjects who are known to cause ciliopathies or subjects who are at risk of developing ciliopathies due to mutations in the subjects' families.In one embodiment of the method described herein, the subject is diagnosed as at risk of developing said ciliopathies, and the method prevents or delays the onset and / or development of ciliopathies in the subject.In an embodiment, the subject is diagnosed as at risk of developing said ciliopathies because they have mutations in the genes described herein.

[0096] In a second aspect, the invention provides a method (Method 2) for treating a disease or disorder selected from obesity, liver disease, retinal degeneration, olfactory dysfunction, hyperlipidemia, type 2 diabetes, and metabolic syndrome in a subject suffering from a ciliopathy, comprising administering to the subject an effective amount of a quinuclidine compound as 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. There is also provided a quinuclidine compound as described herein, e.g., a compound according to Formulas I or II-XII, Ia-XIIa, or Ib-XIIb, or compounds 1 or any of 1.1-1.75, for use in a method for treating a disease or disorder selected from obesity, liver disease, retinal degeneration, olfactory dysfunction, hyperlipidemia, type 2 diabetes, and metabolic syndrome in a subject suffering from a ciliopathy, e.g., for use in any of Methods 2 or 2.1-2.61. Further provided is the use of a quinuclidine compound as described herein, e.g., 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, in the manufacture of a medicament for use in a method for treating a disease or disorder selected from obesity, liver disease, retinal degeneration, olfactory dysfunction, hyperlipidemia, type 2 diabetes, and metabolic syndrome in a subject suffering from a ciliopathic disorder, such as in the manufacture of a medicament for use in any of Methods 2 or 2.1-2.61.

[0097] 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 1.1-1.75; 2.4 any of Methods 2 or 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 1.1-1.75; 2.5 Method 2.3 or 2.4, wherein the pharmaceutical composition further comprises at least one pharma- ceutically acceptable excipient as described herein; 2.6 Any of Methods 2 or 2.1-2.5, wherein the method comprises administering a pharmaceutical dosage form comprising an effective amount of the compound or an effective amount of the pharmaceutical composition; 2.7 Method 2.6, where 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, where the dosage form is a chewable tablet; 2.9 Method 2.6, wherein the dosage form is a parenteral dosage form (e.g., the pharmaceutical composition is formulated for injection); 2.10 The injection is intravenous, intramuscular, intrathecal or subcutaneous, optionally a sterile injection, Method 2.9; 2.11 Method 2.6, where 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 through 2.12, further comprising simultaneously administering in a patient in need thereof a second active agent as described herein, e.g., a second compound capable of treating or preventing a ciliopathies; 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, or any of 2.1-2.14, wherein the subject is a mammal; 2.16 The subject is a primate, method 2.15; 2.17 The subject is a human, method 2.16; 2.18 Any of methods 2 or 2.1 to 2.17, wherein the ciliopathy is a disease selected from the group consisting of Joubert syndrome, Meckel-Gruber syndrome, Senior-Loken syndrome, orofacial-digital syndrome type I, Leber congenital amaurosis, Bardet-Biedl syndrome (BBS), Amström syndrome, June asphyxiating thoracic dystrophy, Ellis-van Creveld syndrome, Sensenbrenner syndrome, and primary ciliary dyskinesia, or a combination thereof; 2.19 Ciliopathy is BBS, either method 2 or 2.1-2.18; 2.20 The ciliopathy is Meckel-Gruber syndrome, method 2.18 or 2.19; 2.21 The ciliopathy is Senior-Loken syndrome, any of methods 2.18-2.20; 2.22 The ciliopathies are Joubert syndrome, methods 2.18-2.21; 2.23 The ciliopathy is Leber congenital amaurosis, any of methods 2.18 to 2.22; 2.24 The ciliopathy is orofacial-digital syndrome type I, any of methods 2.18-2.23; 2.25 The ciliopathies is Amström syndrome, any of methods 2.18-2.24; 2.26 The ciliopathy is Ellis-van Creveld syndrome, any of methods 2.18-2.25; 2.27 The ciliopathy is Sensenbrenner syndrome, any of methods 2.18-2.26; 2.28 The ciliopathies is primary ciliary dyskinesia, any of methods 2.18-2.27; 2.29 The subject has been diagnosed with a mutation in one or more of the genes BBS1 (ARL), BBS2, BBS3, BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9 (B1), BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (C2ORF86 / FRITZ), BBS16 (SDCCAG8), BBS17, BBS18, BBS19, BBS20, and BBS21, according to any of methods 2 or 2.1-2.28; 2.30 Method 2.29, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1, BBS2, BBS4, BBS5, BBS7, BBS8, BBS9, BBS10, and BBS18; 2.31 Method 2.30, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1, BBS2, and BBS10; 2.32 Method 2.31, wherein the subject has been diagnosed with a mutation in at least gene BBS2; 2.33 Any of methods 2 or 2.1-2.32, wherein the subject has been diagnosed with a mutation in one or more of the genes MKS1, MKS3, CEP290, RPGRIP1L, CC2D2A, and TMEM216; 2.34 Any of methods 2 or 2.1 to 2.32, wherein the subject has been diagnosed with a mutation in at least the gene MKS1; 2.35 The subject has been diagnosed with a mutation in one or more of the following genes: TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, TTC21B, KIF7, TCTN1, TMEM237, CEP41, TMEM138, C5ORF42, TCTN3, ZNF423, TMEM231, CSPP1, ARMC9, INPP5E, CXORF5, INVS, NPHP3, NPHP4, NPHP5(IQCB1), and SDCCAG8; 2.36 Any of methods 2 or 2.1-2.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, TTC21B, KIF7, TCTN1, TMEM237, CEP41, TMEM138, C5ORF42, TCTN3, ZNF423, TMEM231, CSPP1, ARMC9, INPP5E, and CXORF5; 2.37 Any of methods 2 or 2.1-2.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, INPP5E, and CXORF5; 2.38 Method 2 or any of 2.1 to 2.37, wherein the subject has been diagnosed with a mutation in one or more of the genes AHI1, ARL13B, INPP5E, and OFD1. 2.39. Any of methods 2 or 2.1-2.38, wherein the subject has been diagnosed with a mutation in one or more of the genes CEP290, NPHP1, INVS, NPHP3, NPHP4 and NPHP5. 2.40 The subject has been diagnosed with a mutation in gene OFD1, according to any of methods 2 or 2.1 to 2.39; 2.41 The subject has been diagnosed with a mutation in one or more of the genes GUCY2D, RPE65, SPATA7, AIPL1, LCA5, RPGRIPL1, CRX, CRB1, IMPD1, RD3, CEP290, NPHP5, and RDH12, according to any of methods 2 or 2.1-2.40; 2.42 Any of methods 2 or 2.1-2.40, wherein the subject has been diagnosed with a mutation in one or more of the genes GUCY2D, RPE65, SPATA7, AIPL1, LCA5, CRX, CRB1, IMPD1, RD3, and RDH12; 2.43 The subject has been diagnosed with a mutation in the gene ALMS1, according to any of methods 2 or 2.1 to 2.42; 2.44 The subject has been diagnosed with a mutation in gene IFT80, according to any of methods 2 or 2.1 to 2.43; 2.45 Any of methods 2 or 2.1-2.44, wherein the subject has been diagnosed with a mutation in one or more of the genes EVC1, EVC2, IFT122, IFT43, and WDR35; 2.46 Method 2.45, wherein the subject has been diagnosed with a mutation in one or more of the genes IFT122, IFT43 and WDR35; 2.47 Method 2.33, wherein the subject has been diagnosed with a mutation in one or more of genes EVC1 and EVC2; 2.48 Method 2 or any of 2.1-2.47, wherein the subject has been diagnosed with a mutation in one or more of the genes DNAI1, DNAH5, TXNDC3, DNAH11, DNAI2, KTU, RSPH4A, RSPH9, and LRRC50; 2.49 The subject also suffers from a glycolipid storage or accumulation disease or disorder, any of methods 2, or 2.1-2.48; 2.50 The glycolipid storage or accumulation disease or disorder is selected from polycystic kidney disease (PKD) (e.g., autosomal dominant PKD [ADPKD]), gangliosidosis (e.g., GM1 gangliosidosis or GM2 gangliosidosis or GM3 gangliosidosis), Gaucher disease (e.g., Gaucher type 1, Gaucher type 2, or Gaucher type 3), Fabry disease, and Parkinson's disease (e.g., Duchenne Parkinson's disease), method 2.49; 2.51 The subject is also being treated with an enzyme replacement therapy (ERT), e.g., using glucocerebrosidase (e.g., alglucerase, imiglucerase, velaglucerase, or taliglucerase), alpha-galactosidase (e.g., agalsidase alpha or agalsidase beta), or beta-galactosidase, each of which optionally is a recombinant enzyme, Method 2, or any of 2.1-2.49; 2.52 Method 2, or any of 2.1-2.51, wherein the subject is administered a daily dose of a compound from about 1 mg to about 150 mg, e.g., selected from 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; 2.53 Any of Methods 2, or 2.1-2.52, wherein the subject is a human pediatric patient, e.g., between 0 and 18 years of age, e.g., between 1 and 15 years of age, or between 1 and 5 years of age, or between 5 and 10 years of age, or between 10 and 15 years of age; 2.54 Method 2, or any of 2.1-2.53, wherein the method is effective to treat, reduce, or ameliorate one or more symptoms or signs selected from obesity, liver disease (e.g., elevated serum liver enzymes, e.g., ALT, AST, alkaline phosphatase, gamma glutamyl transpeptidase), retinal degeneration, hyperlipidemia (e.g., elevated serum total cholesterol, LDL, VLDL, or triglycerides), type 2 diabetes (e.g., elevated serum glucose), and olfactory dysfunction; 2.55 any of Methods 2, or 2.1-2.54, wherein the method is effective to preserve or improve ciliary function in the hypothalamus, retina and / or olfactory epithelium, e.g., to preserve or improve cilia function (e.g., motility) and / or to preserve or improve the quantity or density of functioning cilia; 2.56 Any of Methods 2, or 2.1-2.55, wherein the compound, or a pharma- ceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a parenteral or non-parenteral route; 2.57 The route of administration is oral (enteral), Method 2.56; 2.58 Method 2.56, in which the route of administration is parenteral, e.g., by injection, e.g., by intravenous injection; 2.59 Any of Methods 2, or 2.1-2.55, wherein the compound, or a pharma- ceutically acceptable salt or prodrug thereof, is administered by localized administration, e.g., by topical administration; 2.60 The compound is (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate, method 2, or any of 2.1-2.59; 2.61 Method 2, or any of 2.1-2.60, 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.

[0098] In a third aspect, the invention provides a method for preserving or improving ciliary function in a subject in need thereof, optionally a subject having a ciliopathic disease (Method 3), comprising administering to the subject an effective amount of a quinuclidine compound as described herein, such as a compound according to formula I or any of II-XII, Ia-XIIa or Ib-XIIb, or compounds 1 or any of 1.1 to 1.75. There is also provided a quinuclidine compound as described herein, such as a compound according to formula I or any of II-XII, Ia-XIIa or Ib-XIIb, or compounds 1 or any of 1.1 to 1.75, for use in a method for preserving or improving ciliary function in a subject in need thereof, for example for use in any of Methods 3 or 3.1 to 3.62. Further provided is the use of a quinuclidine compound as described herein, e.g., 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, in the manufacture of a medicament for use in a method for preserving or improving ciliary function in a subject in need thereof, e.g., in the manufacture of a medicament for use in any of methods 3 or 3.1-3.62.

[0099] 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 Compound 1 or any of 1.1 to 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 to 1.75; 3.3 any of Methods 3 or 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 Compound 1 or any of 1.1-1.75; 3.4 Any of Methods 3 or 3.1-3.2, comprising administering to a subject an effective amount of a pharmaceutical composition comprising Compound 1, or any one or more of Compounds 1.1-1.75; 3.5 Method 3.3 or 3.4, wherein the pharmaceutical composition further comprises at least one pharma- ceutically acceptable excipient as described herein; 3.6 Any of Methods 3 or 3.1-3.5 comprising administering an effective amount of a compound or a pharmaceutical dosage form comprising an effective amount of a pharmaceutical composition; 3.7 Method 3.6, where 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, where the dosage form is a chewable tablet; 3.9 Method 3.6, wherein the dosage form is a parenteral dosage form (e.g., the pharmaceutical composition is formulated for injection); 3.10 The injection is intravenous, intramuscular, intrathecal or subcutaneous, optionally a sterile injection, Method 3.9; 3.11 Method 3.6, where the dosage form is a topical or rectal dosage form; 3.12 Method 3.6, where the dosage form is an intranasal dosage form (e.g., an aerosol); 3.13 Any of Methods 3 or 3.1 through 3.12, further comprising simultaneously administering in a patient in need thereof a second active agent as described herein, e.g., a second compound capable of treating or preventing a ciliopathies; 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 Method 3, or any of 3.1-3.14, wherein the subject is a mammal; 3.16 Method 3.15, in which the subject is a primate; 3.17 The subject is a human, method 3.16; 3.18 Any of Methods 3 or 3.1-3.17, wherein the subject has a ciliary disease, e.g., a disease selected from the group consisting of Joubert syndrome, Meckel-Gruber syndrome, Senior-Loken syndrome, Orofacial-Digital syndrome type I, Leber congenital amaurosis, Bardet-Biedl syndrome (BBS), Amstrom syndrome, June asphyxiating thoracic dystrophy, Ellis-van Creveld syndrome, Sensenbrenner syndrome, and primary ciliary dyskinesia, or a combination thereof; 3.19 Ciliopathy is BBS, method 3.18; 3.20 The ciliopathy is Meckel-Gruber syndrome, method 3.18 or 3.19; 3.21 The ciliopathy is Senior-Loken syndrome, any of methods 3.18-3.20; 3.22 The ciliopathy is Joubert syndrome, any of methods 3.18-3.21; 3.23 The ciliopathy is Leber congenital amaurosis, any of methods 3.18 to 3.22; 3.24 The ciliopathy is orofacial-digital syndrome type I, any of methods 3.18-3.23; 3.25 The ciliopathies is Amström syndrome, any of methods 3.18-3.24; 3.26 The ciliopathy is Ellis-van Creveld syndrome, any of methods 3.18-3.25; 3.27 The ciliopathy is Sensenbrenner syndrome, any of methods 3.18-3.26; 3.28 The ciliopathies is primary ciliary dyskinesia, any of methods 3.18-3.27; 3.29 Any of methods 3 or 3.1-3.28, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1 (ARL), BBS2, BBS3, BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9 (B1), BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (C2ORF86 / FRITZ), BBS16 (SDCCAG8), BBS17, BBS18, BBS19, BBS20, and BBS21; 3.30 Method 3.29, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1, BBS2, BBS4, BBS5, BBS7, BBS8, BBS9, BBS10, and BBS18; 3.31 Method 3.30, wherein the subject has been diagnosed with a mutation in one or more of genes BBS1, BBS2, and BBS10; 3.32 Method 3.31, wherein the subject has been diagnosed with a mutation in at least gene BBS2; 3.33 Any of methods 3 or 3.1-3.32, wherein the subject has been diagnosed with a mutation in one or more of the genes MKS1, MKS3, CEP290, RPGRIP1L, CC2D2A, and TMEM216; 3.34 Any of methods 3 or 3.1-3.32, wherein the subject has been diagnosed with a mutation in at least the gene MKS1; 3.35 The subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, TTC21B, KIF7, TCTN1, TMEM237, CEP41, TMEM138, C5ORF42, TCTN3, ZNF423, TMEM231, CSPP1, ARMC9, INPP5E, CXORF5, INVS, NPHP3, NPHP4, NPHP5(IQCB1), and SDCCAG8; 3.36 Method 3 or any of 3.1-3.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, TTC21B, KIF7, TCTN1, TMEM237, CEP41, TMEM138, C5ORF42, TCTN3, ZNF423, TMEM231, CSPP1, ARMC9, INPP5E, and CXORF5; 3.37 Any of methods 3 or 3.1-3.34, wherein the subject has been diagnosed with a mutation in one or more of the genes TMEM216, AHI1, NPHP1, CEP290, TMEM67, RPGRIP1L, ARL13B, CC2D2A, INPP5E, and CXORF5; 3.38 Method 3 or any of methods 3.1 to 3.37, wherein the subject has been diagnosed with a mutation in one or more of the genes AHI1, ARL13B, INPP5E, and OFD1. 3.39. Any of methods 3 or 3.1-3.38, wherein the subject has been diagnosed with a mutation in one or more of the genes CEP290, NPHP1, INVS, NPHP3, NPHP4 and NPHP5. 3.40 The subject has been diagnosed with a mutation in gene OFD1, according to any of methods 3 or 3.1 to 3.39; 3.41 any of methods 3 or 3.1-3.40, wherein the subject has been diagnosed with a mutation in one or more of the genes GUCY2D, RPE65, SPATA7, AIPL1, LCA5, RPGRIPL1, CRX, CRB1, IMPD1, RD3, CEP290, NPHP5, and RDH12; 3.42 Any of methods 3 or 3.1-3.40, wherein the subject has been diagnosed with a mutation in one or more of the genes GUCY2D, RPE65, SPATA7, AIPL1, LCA5, CRX, CRB1, IMPD1, RD3, and RDH12; 3.43 The subject has been diagnosed with a mutation in the gene ALMS1, according to any of methods 3 or 3.1 to 3.42; 3.44 The subject has been diagnosed with a mutation in gene IFT80, any of methods 3 or 3.1-3.43; 3.45 Any of methods 3 or 3.1-3.44, wherein the subject has been diagnosed with a mutation in one or more of the genes EVC1, EVC2, IFT122, IFT43, and WDR35; 3.46 Method 3.45, wherein the subject has been diagnosed with a mutation in one or more of genes IFT122, IFT43 and WDR35; 3.47 Method 3.33, wherein the subject has been diagnosed with a mutation in one or more of genes EVC1 and EVC2; 3.48. Any of methods 3 or 3.1-3.47, wherein the subject has been diagnosed with a mutation in one or more of the genes DNAI1, DNAH5, TXNDC3, DNAH11, DNAI2, KTU, RSPH4A, RSPH9, and LRRC50; 3.49 The subject also suffers from a glycolipid storage or accumulation disease or disorder, any of Methods 3, or 3.1-3.48; 3.50 The glycolipid storage or accumulation disease or disorder is selected from polycystic kidney disease (PKD) (e.g., autosomal dominant PKD [ADPKD]), gangliosidosis (e.g., GM1 gangliosidosis or GM2 gangliosidosis or GM3 gangliosidosis), Gaucher disease (e.g., Gaucher type 1, Gaucher type 2, or Gaucher type 3), Fabry disease, and Parkinson's disease (e.g., Duchenne Parkinson's disease), Method 3.49; 3.51 The subject is also being treated with an enzyme replacement therapy (ERT), e.g., using glucocerebrosidase (e.g., alglucerase, imiglucerase, velaglucerase, or taliglucerase), alpha-galactosidase (e.g., agalsidase alpha or agalsidase beta), or beta-galactosidase, each of which optionally is a recombinant enzyme, Method 3, or any of 3.1-3.49; 3.52 Method 3, or any of 3.1-3.51, wherein the subject is administered a daily dose of a compound from about 1 mg to about 150 mg, e.g., selected from 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; 3.53 Any of Methods 3, or 3.1-3.52, wherein the subject is a human pediatric patient, e.g., between 0 and 18 years of age, e.g., between 1 and 15 years of age, or between 1 and 5 years of age, or between 5 and 10 years of age, or between 10 and 15 years of age; 3.54 The subject has a comorbidity selected from obesity, liver disease, retinal degeneration, olfactory defects, hyperlipidemia, type 2 diabetes, and metabolic syndrome, method 3, or any of 3.1-3.53; 3.55 Method 3, or any of 3.1-3.54, wherein the method is effective to treat, reduce, or ameliorate one or more symptoms or signs selected from obesity, liver disease (e.g., elevated serum liver enzymes, e.g., ALT, AST, alkaline phosphatase, gamma glutamyl transpeptidase), retinal degeneration, hyperlipidemia (e.g., elevated serum total cholesterol, LDL, VLDL, or triglycerides), type 2 diabetes (e.g., elevated serum glucose), and olfactory dysfunction; 3.56 Any of Methods 3, or 3.1-3.55, wherein the method is effective to preserve or improve ciliary function in the hypothalamus, retina and / or olfactory epithelium, e.g., to preserve or improve function (e.g., motility) of cilia and / or to preserve or improve the quantity or density of functioning cilia; 3.57 Method 3, or any of 3.1-3.56, wherein the compound, or a pharma- ceutically acceptable salt or prodrug thereof, is administered by systemic administration, e.g., via a parenteral or non-parenteral route; 3.58 The route of administration is oral (enteral), Method 3.57; 3.59 Method 3.57, in which the route of administration is parenteral, e.g., by injection, e.g., by intravenous injection; 3.60 Any of Methods 3, or 3.1-3.56, wherein the compound, or a pharma- ceutically acceptable salt or prodrug thereof, is administered by localized administration, e.g., by topical administration; 3.61 Method 3, or any of 3.1-3.60, wherein the compound is (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate; 3.62 Method 3, or any of methods 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.

[0100] Pharmaceutical Compositions The present disclosure also provides pharmaceutical compositions comprising at least one quinuclidine compound described herein and at least one pharma- ceutically acceptable excipient, for example, for use according to the methods disclosed herein.The pharma-ceutically acceptable excipient may be any excipient known in the art, including, for example, those described in Remington's Pharmaceutical Sciences, Mack Publishing Co. (AR Gennaro edit. 1985).The 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 pharma-ceutically acceptable excipient.

[0101] Thus, in one embodiment, the invention provides a pharmaceutical dosage form comprising a quinuclidine compound described herein and a pharma- ceutically acceptable excipient, the dosage form being formulated to provide a sufficient amount of the compound when administered (e.g., when administered orally) to treat a ciliopathic disorder.

[0102] The pharmaceutical compositions or dosage forms of the present invention may contain a drug and another carrier, such as an inactive or active compound or composition, such as a detectable agent, a label, an adjuvant, a diluent, a binder, a stabilizer, a buffer, a salt, a lipophilic solvent, a preservative, an adjuvant, 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, such as alditols, aldonic acids, esterified sugars, and the like; 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, such as 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, examples of which include, but are 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.

[0103] 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 salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid; Tris, tromethamine hydrochloride, or phosphate buffers. Additional carriers include polymer excipients / additives, such as polyvinylpyrrolidone, ficoll (polymeric sugar), dextrates (e.g., cyclodextrins, such as 2-hydroxypropyl-β-cyclodextrin), polyethylene glycol, flavoring agents, antibacterial agents, sweeteners, antioxidants, antistatic agents, surfactants (e.g., polysorbates, such as "TWEEN 20" and "TWEEN 80"), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol), and chelating agents (e.g., EDTA).

[0104] The present disclosure also provides pharmaceutical compositions and kits comprising said compositions, comprising at least one quinuclidine compound and at least one additional pharmacoactive agent as described herein.These pharmaceutical compositions and kits may be adapted to allow simultaneous, sequential and / or separate administration of quinuclidine compound and additional active agent.For example, quinuclidine compound and additional active agent may be formulated in separate dosage forms, for example, separate tablets, capsules, lyophilisates or liquids, or in the same dosage form, for example, the same tablets, capsules, lyophilisates or liquids.When quinuclidine compound and additional active agent are formulated in the same dosage form, quinuclidine compound and additional active agent may be substantially present in additive mixture, for example, in the core of a tablet, or in separate regions of the dosage form, for example, in separate layers of the same tablet.In one embodiment, the pharmaceutical dosage form comprises additional agents that can treat or prevent ciliopathies, for example, ciliopathies as described herein.

[0105] In a further aspect, the present invention provides a pharmaceutical composition comprising: (i) a quinuclidine compound as described herein; (ii) an additional active agent; and (iii) a pharmaceutically acceptable excipient.In one embodiment, the additional active agent is an agent that can treat or inhibit a ciliopathy, for example, a ciliopathy as described herein.In one embodiment, the additional active agent can treat or inhibit a ciliopathy when orally administered to a subject.

[0106] Examples of further drugs that can treat proteinopathies, such as Parkinson's disease, include, for example, dopamine precursors (e.g., L-dopa), dopamine agonists (e.g., bromocriptine, cabergoline, pergolide, pramipexole and apomorphine), MAO-B inhibitors (e.g., rasagiline and selegiline), anticholinergics (e.g., orphenadrine, procyclidine and trihexyphenidyl), β-glucocerebrosidase activity enhancers (e.g., ambroxol and afegostat) and amantadine.Examples of drugs that can treat Alzheimer's disease include, for example, acetylcholinesterase inhibitors, such as tacrine, rivastigmine, galantamine, donepezil and memantine.

[0107] Additional treatments for proteinopathies that may be combined with the methods described herein include psychosocial interventions, behavioral interventions, reminiscence therapy, validation therapy, supportive psychotherapy, sensory integration, cognitive retraining, rehabilitation, speech therapy, etc. Other interventions include surgery, rehabilitation, and dietary management.

[0108] 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, e.g., via a route other than parenteral. In one embodiment, the quinuclidine compounds or pharmaceutical compositions are formulated for oral administration, e.g., 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).

[0109] The pharmaceutical composition may be formulated to delay, extend or control the release of the active ingredient therein, for example, by using various ratios of hydroxypropylmethylcellulose, other polymer matrices, liposomes and / or microspheres to provide the desired release profile. The pharmaceutical composition may also optionally contain opacifying agents, and may optionally be a composition that releases only or preferentially in a certain part of the gastrointestinal tract in a delayed manner, for example, by using an enteric coating. Examples of embedding compositions include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, where appropriate, with one or more pharma- ceutically acceptable carriers, excipients, or diluents known in the art (see, for example, Remington's). The compounds disclosed herein may 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.

[0110] 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 pharma- ceutically 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, alginates, gelatin, pregelatinized maize starch, polyvinylpyrrolidone, hydroxypropylmethylcellulose, sucrose, and / or acacia; (3) humectants, such as glucose, glyceryl stearate, glyceryl stearate, and / or glyceryl stearate; (4) humectants, such as glyceryl stearate, glyceryl stearate, and / or glyceryl stearate; (5) humectants, such as glyceryl stearate, glyceryl stearate, and / or glyceryl stearate; (6) humectants, such as glyceryl stearate, glyceryl stearate, and / or glyceryl stearate; (7) humectants, such as glyceryl stearate, glyceryl stearate, and / or glyceryl stearate; glycerol;(4) disintegrants such as agar-agar, calcium carbonate, sodium starch glycolate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate;(5) dissolution retarders such as paraffin;(6) absorption enhancers 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 glycols, sodium lauryl sulfate, and mixtures thereof; and(10) mixed with coloring agents. In the case of capsules, tablets, and pills, the pharmaceutical composition may also contain buffering agents. Similar types of solid compositions may also be prepared in soft and hard filled gelatin capsules using fillers and excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols.

[0111] Tablets may be prepared 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 prepared 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 be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the art.

[0112] In an embodiment, pharmaceutical compositions are orally administered in liquid form.Liquid dosage forms for oral administration of active ingredients include pharma- ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.Liquid preparations for oral administration may be present as dry products, for which they are made up with water or other suitable vehicles before use.In addition to active ingredients, liquid dosage forms may 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 the inert diluent, 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 pharma-ceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, methylcellulose or hydrogenated edible fats); emulsifying agents (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-hydroxybenzoates or sorbic acid). The active ingredient may also be administered as a bolus, electuary, or paste.

[0113] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0114] In embodiments, the pharmaceutical compositions are administered by parenteral means, e.g., topical application, transdermal application, injection, etc. In related embodiments, the pharmaceutical compositions are administered parenterally by injection, infusion, or implantation (e.g., intravenous, intramuscular, intraarterial, subcutaneous, etc.).

[0115] The compounds disclosed herein may be formulated for parenteral administration by injection, including using conventional catheter techniques or infusion. The formulations for injection may be in unit dosage form, for example, in ampoules or in multi-dose containers, with the addition of preservatives. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulating 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, for example, sterile pyrogen-free water, before use.

[0116] 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 avoiding 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 the 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 one 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).

[0117] The pharmaceutical compositions may be in the form of sterile injections. The pharmaceutical compositions may be rendered sterile, for example, by filtration through a bacteria-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 prior to 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. The pharmaceutical compositions may also contain one or more preservatives, for example, methyl, ethyl, or n-propyl p-hydroxybenzoate. To improve solubility, a solution enhancer or solubilizer may be added, or the solvent may contain 10-60% w / w propylene glycol or similar.

[0118] The pharmaceutical compositions may comprise one or more pharma- ceutically acceptable sterile isotonic aqueous or nonaqueous 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; bacteriostatic agents; solutes that render the formulation isotonic with the blood of the intended recipient; suspending agents; thickening agents; preservatives; and the like.

[0119] 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, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity may 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, in order to prolong the effect of the active ingredient, it is desirable to delay the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the active ingredient then depends on its rate of dissolution, which may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered active ingredient is accomplished by dissolving or suspending the compound in an oil vehicle. In addition, sustained absorption of an injectable pharmaceutical form may be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0120] The controlled release parenteral composition may be in the form of an aqueous suspension, microspheres, microcapsules, magnetic microspheres, oil solutions, oil suspensions, emulsions, or the active ingredient may be incorporated into a biocompatible carrier, liposomes, nanoparticles, implants, or infusion devices. Materials for use in the preparation of microspheres and / or microcapsules include, but are not limited to, biodegradable / bioerodible polymers, such as polyglactin, poly-(isobutylcyanoacrylate), 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).

[0121] For topical administration, the compounds disclosed herein may be formulated into ointments or creams.The compounds disclosed herein may also be formulated into rectal compositions, such as suppositories or retention enemas, for example, containing conventional suppository bases such as cocoa butter or other glycerides.

[0122] For intranasal administration or administration by inhalation, 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, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of pressurized aerosol, the dosage unit 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, such as lactose or starch.

[0123] Generally, the reagents and compositions described herein are administered in an effective amount or amounts sufficient to treat or prevent ciliopathies in subjects.Typically, the dosage may 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 capabilities of those skilled in the art.

[0124] The proposed dose of the quinuclidine compounds described herein for oral, parenteral or buccal administration to an average adult to treat ciliopathies is about 0.1 mg to about 2000 mg. In certain embodiments, the proposed dose is about 0.2 mg to about 1000 mg of active ingredient per unit dose. Regardless of the amount of the proposed dose, administration of the compound may occur, for example, 1 to 4 times a day. In one embodiment, the dose for oral administration is about 0.5 to about 2000 mg, for example, about 1 to about 750 mg. In one embodiment, the dose for direct administration to the central nervous system is about 1 μg to about 1 mg, for example, about 5 μg to about 0.5 mg, or about 10 μg to about 0.1 mg. Aerosol formulations for treating or preventing the above conditions in an average adult may be prepared so that each metered dose or "puff" of aerosol contains about 1 mg to about 10 g, for example, about 2 mg to about 1 g, of the compound disclosed herein. Administration may be several times a day, for example 2, 3, 4 or 8 times, for example giving 1, 2 or 3 doses each time. In some embodiments, administration may be by a single daily dose of 5 mg, 10 mg, 15 mg or 20 mg. In some embodiments, administration may be by a single daily dose of 2, 5, 15, 25, 50, 100 or 150 mg.

[0125] In another aspect, the invention provides a dosage form or pharmaceutical composition as described herein for use in therapy, e.g. for use in a method defined herein.

[0126] Having been generally described herein, the following non-limiting examples are provided to further illustrate the invention. EXAMPLES

[0127] 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 (concentration about 0.2 M) at room temperature was added triphosgene (0.35 equiv.). 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.

[0128] To a solution of the alcohol (1.5 eq.) in THF (concentration about 0.2 M) at room temperature was added NaH [60%, oil] (1.5 eq.). 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.

[0129] General Procedure B: Alkylation with Organocerium A suspension of CeCl3 (4 eq.) in THF (concentration about 0.2 M) was stirred at room temperature for 1 h. The suspension was cooled to -78°C and MeLi / ether [1.6 M] (4 eq.) was added dropwise. The organocerium complex was allowed to form for 1 h and a solution of the nitrile (1 eq.) in THF (concentration 2.0 M) 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 (about 1 mL), followed by the addition of 50% aqueous ammonium hydroxide (about 3 mL) until a precipitate formed and sank to the bottom of the flask. The mixture was filtered through a Celite pad and concentrated. The crude material was treated with HCl / dioxane [4.0 M] solution. The intermediate arylpropan-2-amine hydrochloride was triturated in ether and used directly in the next step. Alternatively, the crude free-based amines were purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding arylpropylamines.

[0130] General Procedure C: Suzuki Coupling To a solution of aryl halide (1 equiv.) in a mixture of DME / water [4:1] (concentration ca. 0.2 M) 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 plug of Celite and concentration, the crude product was purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding coupling adduct.

[0131] Alternative method: To a solution of aryl halide (1 equiv.) in a mixture of toluene / water [20:1] (concentration about 0.2 M) was added 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.). The reaction mixture was microwaved at 150 °C for 25 min. After filtration through a plug of Celite and concentration, the crude product was purified by Combiflash (SiO2 cartridge, CHCl3 and 2N NH3 in MeOH) to give the corresponding coupling adduct.

[0132] General Procedure D: Cyclopropanation To a mixture of aryl nitrile (1 equiv.) and Ti(Oi-Pr)4 (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. To the above mixture, BF3·Et2O (3 equiv.) was added dropwise at 25°C. After the addition, the mixture was stirred for another 2 h and then quenched with aqueous HCI [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 Na2SO4, filtered and concentrated. The crude material was purified by silica gel column chromatography (eluted with petroleum ether / EtOAc: 10 / 1 to 1 / 1) to give the corresponding 1-aryl-cyclopropanamine.

[0133] General Procedure E: Biaryl Coupling Using Suzuki Conditions To a stirred solution of aryl halide component (1 equiv.) in 5:1 (v / v) dioxane / water (~0.15 M) or 5:1 (v / v) N,N-dimethylformamide (~0.15 M) was added aryl boronate or aryl boronic acid component (1-1.5 equiv.), sodium carbonate (2-3 equiv.) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.05 equiv.). 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.

[0134] 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 eq) in tetrahydrofuran (ca. 0.1 M) was added triethylamine (2 eq). The reaction was cooled (0° C.) and treated with isobutyl chloroformate (1.5 eq). After 1 h at 0° C., a solution of sodium azide (2 eq) in water (ca. 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 (ca. 0.1 M). The stirred solution was refluxed for 2-2.5 h, cooled and treated with the alcohol component (1.25-2 eq). 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 products were purified by flash chromatography on silica using chloroform / methanol (less polar carbamates) or chloroform / methanol / ammonia (more polar carbamates) solvent gradients. EXAMPLES

[0135] 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 afforded 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.

[0136] (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 partitioned between ethyl acetate (200 mL) and aqueous NaHCO3 (200 mL). The organic layer was combined with a back-extraction of the aqueous layer (ethyl acetate, 1 x 75 mL), dried (Na2SO4), 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 (13.58 g, 89%).

[0137] 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 min, 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 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 (Na2SO4) and concentrated. The resulting amber oil was purified by flash chromatography using a hexanes / ethyl acetate gradient to give ethyl 2-(2-(4-fluorophenyl)thiazol-4-yl)-2-methylpropanoate as a colorless oil (4.57 g, 66%).

[0138] 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 addition of 1.0 N HCl (80 mL), and extracted with ethyl acetate (2×70 mL). The combined extracts were dried (Na2SO4) 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.

[0139] 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 hour 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 NaHCO3 (1×150 mL) and brine (1×100 mL), dried (Na2SO4), and concentrated. After coevaporation with toluene (2 x 50 mL), the resulting white solid was taken up in toluene (100 mL) and refluxed for 4 h. (S)-3-quinuclidinol (3.87 g, 30.4 mmol) was then added and reflux continued overnight. The reaction was concentrated and the residue was partitioned between ethyl acetate (100 mL) and aqueous NaHCO3 (150 mL). The organic layer was washed with water (1 x 150 mL), dried (Na2SO4) 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.

[0140] (S)-Quinuclidin-3-yl (2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 3) Using general procedure E and using ethyl 2-(4-bromophenyl)-2-methylpropanoate and 4-(2-methoxyethoxy)phenylboronic acid as inputs in the reaction, ethyl 2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)-2-methylpropanoate 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 (Na2SO4) 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 yield 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.

[0141] 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 afforded 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.

[0142] (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.

[0143] 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.

[0144] Using general procedure C, the above bromide (200 mg, 0.540 mmol), phenylboronic acid (133 mg, 1.10 mmol) and [PdCl2(pddf)]CH2Cl2 afforded 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.

[0145] 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.

[0146] 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.

[0147] Using general procedure C, the above carbamate (400 mg, 1.12 mmol), phenylboronic acid (267 mg, 2.22 mmol) and [PdCl2(pddf)]CH2Cl2 afforded 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.

[0148] (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 afforded 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.

[0149] (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)2 (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.

[0150] 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)2 (0.016 g, 0.071 mmol) gave the title compound as a grey solid (0.337 mg, 65%). 1H 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.

[0151] 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-bromothiophene-3-yl)propan-2-ol as a light amber oil (8.05 g, 64%).

[0152] 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 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 NaHCO3 (1×250 mL), dried (Na2SO4), 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 NH4OH and extracted with ethyl acetate (2 x 100 mL). These extracts were dried (Na2SO4) and concentrated. The resulting amber oil was purified by flash chromatography using a methylene chloride / methanol / ammonia gradient to give 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%).

[0153] 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). 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 NaHCO3 (1 x 150 mL), water (2 x 150 mL). The organic layer was dried (Na2SO4) and concentrated to give the crude 4-nitrophenyl quinuclidin-3-yl carbonate product, which was used in the next step without purification.

[0154] To a stirred solution of 2-(5-bromothiophen-3-yl)propan-2-amine (0.366 g, 1.66 mmol) in THF (10 mL) was added 4-nitrophenylquinuclidin-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 NaHCO3 (50 mL). The organic layer was washed again with aqueous NaHCO3 (1 x 50 mL), dried (Na2SO4), 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%).

[0155] 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) afforded the title compound as a grey 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. 13C NMR (400 MHz, CDCl3) δ 162.3 (d, J = 247.1 Hz), 154.5, 149.8, 143.6, 130.7, 127.4 (d, J = 8.1 Hz), 121.8, 118.9, 115.8 (d, J = 21.6 (Hz), 70.8, 55.5, 53.4, 47.3, 46.4, 29.0, 25.4, 24.4, 19.4 ppm. Purity: 95.8 % UPLCMS (210 nm & 254 nm); retention time 0.90 min; (M+1) 389.

[0156] (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 [ca. 1.9 M] (10.8 mL, 20.5 mmol). The reaction was heated to reflux for 2 h 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. 13C 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.

[0157] (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-extraction 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)thiazol-2-yl)acetate as an off-white solid (29.92 g, 83%).

[0158] 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 t-butoxide in THF [1.0 M] (136 mL, 136 mmol) followed by 18-crown-6 (1.6 mL, 7.5 mmol). After a further 30 min at -78°C, iodomethane (8.5 mL) was added dropwise over 5 min. The reaction was stirred cold for a further 2 h before being poured into water (450 mL) and extracted with ethyl acetate (2 x 150 mL). The combined extracts were washed with brine (1 x 200 mL), dried (Na2SO4) 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%).

[0159] 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 addition of 1N HCl (15 mL), and extracted with ethyl acetate (2×50 mL). The combined extracts were dried (Na2SO4) and concentrated to give 2-(4-(4-fluorophenyl)thiazol-2-yl)-2-methylpropanoic acid as a pale gold solid (0.808 g, 99%).

[0160] 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 hour 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 NaHCO3 (1×150 mL) and brine (1×100 mL), dried (Na2SO4), and concentrated. After coevaporation with toluene (2×30 mL), the resulting off-white solid was taken up in toluene (25 mL) and refluxed for 4 h. (S)-3-quinuclidinol (0.753 g, 5.92 mmol) was then added and reflux continued for 3 h. 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.

[0161] 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 produce the title compound as a colorless glassy solid (23%). NMR data was consistent with that of Example 3. Purity: 100%, 99.1% (210 and 254 nm) UPLCMS; Retention time: 0.87 min; (M+H + )439.0.

[0162] (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. 1H 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.

[0163] 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. 1H 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.

[0164] 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 (Na2SO4) 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 1N 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 as a waxy off-white solid (0.630 g, 99%). This intermediate and quinuclidin-3-ol were reacted according to general procedure F to yield 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.

[0165] 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 light 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 chloroform / methanol / ammonia eluent afforded 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.

[0166] Quinuclidin-3-yl (2-(4'-(2-hydroxyethyl)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate (Compound 18) Using general procedure E and employing 1-(2-(benzyloxy)ethyl)-4-bromobenzene and ethyl 2-methyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate in the reaction, ethyl 2-(4'-(2-(benzyloxy)ethyl)-[1,1'-biphenyl]-4-yl)-2-methylpropanoate was prepared 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 part 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 1.0 N hydrochloric acid (1 mL) and 10% palladium on carbon (50% water; 0.087 g). 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. 1H 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.

[0167] 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 h, 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 h, 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 min. The resulting red suspension was cooled (0° C.) and treated dropwise with iodomethane (9.80 mL, 157 mmol) over 10 min. The cooling bath was removed and the reaction was stirred for 4 h, then concentrated and the residue partitioned between ethyl acetate and water. The organic layer was washed two more times 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 light 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 min. After stirring overnight, the reaction was quenched by 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 h, 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 (Na2SO4), 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 partitioned between water and diethyl ether. The aqueous layer was treated with 1.0 N hydrochloric acid (40 mL) and extracted with ethyl acetate. The combined extracts were dried (Na2SO4) 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. 13C 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.

[0168] 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 2 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 (Na2SO4) 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.

[0169] 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, using 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 before adding a solution of the above arylborate (1.83 g, 7.20 mmol) in tetrahydrofuran (20 mL). 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 (Na2SO4) and concentrated. The residue was purified by flash chromatography on silica using ethyl acetate eluent to give 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) for 1 h. After the addition, the reaction was stirred for an additional 30 minutes before removing the top toluene layer and drying (Na2SO4). 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 minutes and then added to the crude isocyanate solution in toluene. The reaction was stirred for 10 minutes, quenched by the addition of brine (5 mL) and extracted with ethyl acetate.The combined extracts were dried (Na2SO4) 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%). 1 H 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.

[0170] 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 (Na2SO4), 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 to 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. 13C 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.

[0171] 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. EXAMPLES

[0172] 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 RB flask was equipped with a thermometer, addition funnel and 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 the powder funnel followed by THF (60 mL). A cloudy solution resulted when most of the potassium tert-butoxide had 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 as a 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 min, maintaining the internal temperature below 10 °C throughout the addition. After the addition was complete, the slightly cloudy mixture was stirred for an additional 1 h, during which time the internal temperature dropped to 0-5 °C. After stirring at 0-5 °C for 1 h, the reaction mixture was quenched by the slow dropwise addition of 5.0 M aqueous HCl (8 mL) over 5-7 min. 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 min. Stirring was stopped 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 vacuum 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.

[0173] Step 2: Hydrolysis of the ethyl ester using LiOH monohydrate [ka] The crude ester in THF was added to the reaction flask. Separately, LiOH.H2O (MW 41.96, 75.0 mmol, 3.15 grams, 2.2 equiv.) was weighed into a 100 mL beaker to which a stir bar was added. Water (40 mL) was added and the mixture was stirred until all solids were dissolved to give 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 cooled to room temperature. The THF was evaporated in vacuo to give a brown solution. An aliquot of the brown aqueous solution was analyzed by HPLC and LC / MS to indicate complete hydrolysis of the ethyl ester. Water (15 mL) was added and the aqueous basic solution was extracted with TBME (2 x 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 with stirring to a pH of approximately 1. TBME (60 mL) was added to this sticky solid in the aqueous acidic solution and the mixture was shaken and then vigorously stirred to dissolve all 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 aforementioned 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 and to give the crude acid as an orange / dark yellow oil which was 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 give the acid as a yellow solid.

[0174] Step 3: Formation of hydroxamic acid using 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. The acid readily dissolved upon addition of THF (5.0 mL) to give 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 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 as this 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 an 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 vacuum and the residue was taken up in dichloromethane (120 mL) and water (60 mL). The mixture was transferred to a separatory funnel, which was 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.

[0175] Continuation of step 3: 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.

[0176] 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 CO2 and rearranged to the isocyanate (see below). [ka]

[0177] 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 as a solid to the mixture in one portion. The mixture was again heated to reflux for 18 h. After 18 h, an aliquot was analyzed by HPLC and LC / MS, which showed that conversion of the isocyanate to the desired product was complete. 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 out of solution as a dull yellow sticky solid that could be captured with the stir bar. Isopropyl acetate (100 mL) was added to this mixture and the mixture was stirred vigorously for 5 min where the sticky solid transferred to the 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 that was dried under high vacuum overnight.

[0178] Continued from step 3: Recrystallization of the 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 stir bar and the mixture was heated to reflux for 10 minutes (the free base was initially partially dissolved, but when heated to reflux it dissolved and gave 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 on the Buchner funnel with a vacuum hose overnight. 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. EXAMPLES

[0179] 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.

[0180] For example, the free base of (S)-quinuclidin-3-yl (2-(2-(4-fluorophenyl)thiazol-4-yl)propan-2-yl)carbamate (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 give the desired malate salt.

[0181] Other salt crystalline forms, such as acid addition salts with succinic acid or HCl, may be prepared in a similar manner. EXAMPLES

[0182] Effect of Compound 1 on ciliary structure and signaling Mouse model ABbs2 - / - The mouse model was described by Nishimura DY et al. (Proc Natl Acad Sci, 101:16588-16593 (2004)) in which exons 5-14 of Bbs2 were replaced with a neomycin cassette. Mice were backcrossed to the 129 / SvJ background. - / - Mice were fed ad libitum with 0.033% w / w Compound 1 in their diet from 1 to 6 months of age. - / - and Wt control animals were fed a normal 5053 chow diet (LabDiet). - / - Mice were left untreated until the age of 4 months and then treated with Compound 1 incorporated into the diet at 5 and 6 months.

[0183] Cell culture and primary cilia staining Wild type and Bbs2- / - Renal epithelial cell lines were established and maintained as previously described (see Natoli T et al., Nat Med, 16:788-792 (2010); and Humes HD. et al., Am J Kidney Dis, 39:1078-1087 (2002)). Cells were cultured on collagen I-coated glass slides in DMEM with 1% penicillin / streptomycin, 10% FBS. To determine the effect of compounds on cilia and lipid localization, cells were cultured in serum-free medium for 24 hours, followed by addition of compound 1 for 6-24 hours. Cells were then fixed with 4% paraformaldehyde, followed by immunofluorescence with anti-GM3 (Creative Biolabs), anti-ceramide (Sigma Aldrich), anti-GM1 (Invitrogen), and anti-acetylated tubulin (Cell Signaling) antibodies. Cilia length was quantified using Metamorph software.

[0184] Discussion and Results To investigate the mechanism of action of the quinuclidine compounds described herein on ciliary structure and signaling, Wt and Bbs2 - / - We used immortalized kidney epithelial cells from mice. First, we analyzed the effect of the mutation Bbs2 on cilia length. - / - We observed that the cilia in the Bbs2 cells were shorter than those in the WT cells (Fig. 1A). - / - We examined the specific levels of glycosphingolipids (GSLs), GM3, GM1, and ceramide in the cells. Immunofluorescence analysis showed that GM3 and ceramide were localized to the primary cilium (Figure 1B). GM3 was upregulated in Bbs2 cells compared to Wt cells. - / - Compared to Wt mice, cytoplasmic Bbs2 was abundant in the primary cilia of the cells. - / - Increased ceramide levels in cells were also observed. GM1 was not localized to cilia, but was localized in intracellular vesicular compartments (Fig. 1B).

[0185] Finally, Wt and Bbs2 - / -We investigated the effect of treatment with compound 1 on GSL distribution in cells. Treatment with compound 1 had the greatest effect on the distribution of GM3. Treatment elongated cilia and restored GM3 localization to that observed in WT cells (Figure 1C). Taken together, these data suggest that Bbs2 - / - The cells were characterized by short cilia in which GSLs were mislocalized compared to wild-type cells, and it was demonstrated that treatment with quinuclidine compounds, such as Compound 1, could restore cilia length and GM3 localization to similar to that observed in wild-type cells. EXAMPLES

[0186] Preclinical in vivo efficacy studies of compound 1 GL1 level evaluation GL1 analysis Quantitative analysis of glycosylceramides was performed by liquid chromatography and tandem mass spectrometry (LC / MS / MS). Briefly, 100 mg of tissue was homogenized in 1 ml of water using a Mini Beadbeater (BioSpec Products, Inc., Bartlesville, OK). 10 μl of the homogenate was extracted with 1 ml of 90% 96:2:1:1 acetonitrile / methanol / acetic acid / water (v / v / v / v) (mobile phase A) and 10% 98:1:1 methanol / acetic acid / water (v / v / v) (mobile phase B), both containing 5 mM ammonium acetate. Samples were placed on a VX-2500 tube vortex (VWR International, LLC, MA) for 5 min and then centrifuged at 8,400 rpm for 4 min (Beckman Coulter, Inc., IN). The resulting supernatant was transferred to HPLC vials for analysis. Glycosylceramides were collected using an Acquity UPLC (Waters Corp., Milford, MA) interfaced to an AB Sciex API 5000 triple quadrupole mass spectrometer (Applied Biosystems, Foster City, CA). Glucosylceramide (GL1) and galactosylceramide were separated by normal phase LC using a 2.1 mm × 150 mm Waters Atlantis HILIC silica column. Quantitation was performed using a GL1 standard (glucocerebroside, Gaucher spleen; Matreya, LLC, Pleasant Gap, PA).

[0187] Discussion and Results Preclinical in vivo efficacy studies were performed using BBS2 - / - Compound 1 was used in a mouse model. - / - The mouse model reproduces the major clinical features of human BBS, including obesity, retinal degeneration, neurological and skeletal abnormalities, liver findings, and anosmia. To determine the therapeutic benefit of the quinuclidine compounds described herein in treating BBS, Bbs2 - / -Mice were treated with 0.033% w / w Compound 1 in the diet from 1 to 6 months of age. This treatment reduced the levels of GL1 in the brain, kidney, liver and serum, suggesting full target engagement (Figure 2) (*p<0.05). EXAMPLES

[0188] Effect of compound 1 on metabolic parameters body composition Body composition was analyzed using EchoMRI™. Fat mass and lean mass measurements were recorded and body fat percentage was calculated. Body composition was calculated using the following formula: fat mass / (fat mass+lean mass).

[0189] Food consumption Food weight was recorded from the food hopper and bedding. The average food consumption per animal per day was assessed using the following formula: [(weight of food at the start of the period - (weight of food in the hopper at the end of the period - weight of food in the bedding at the end of the period)] / #animals in cage / #number of days of observation.

[0190] Serum leptin Blood was collected during necropsy and incubated at room temperature for 15 minutes to allow clot formation. The clot was removed by centrifugation at 15,000 rpm for 5 minutes to collect serum. Leptin concentrations were measured using a leptin ELISA kit (R&D Systems) according to the manufacturer's instructions.

[0191] Real-time quantitative PCR Total RNA was isolated from dissected, homogenized adipose tissue from 6-month-old mice using TRizol and chloroform extraction followed by RNAeasy Mini Kit purification (Qiagen), quantified using a NanoDrop 2300 system, and reverse transcribed using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Primers for Fas and Srebf1 were purchased from Applied Biosystems. Quantitative PCR was performed in duplicate for each sample on an Applied Biosystems RT-PCR instrument using Taqman Universal Master Mix (Thermo Fisher) according to the manufacturer's instructions. Relative amounts of mRNA were quantified, determined using the comparative CT method, and normalized to GAPDH mRNA levels.

[0192] Calculating cell volume Automated digital image analysis was performed on the whole adipose tissue using Visiopharm Image Analysis software (DK-2970 Hoersholm, Denmark, version 6.9.1). Two custom-made applications were created and run consecutively on each digital image. The first application detected adipose tissue using thresholding and outlined the area as a region of interest (ROI). The second application used thresholding to classify the tissue within the ROI into three categories: cytoplasm (plasma membrane), fat (adipocytes), and other (undesirable artifacts, large blood vessels, other tissues). Post-processing steps included enclosing fat with cytoplasm and selecting appropriate adipocytes to count them by removing any fat with a formation factor below 0.5.

[0193] Immunofluorescence Wt and Bbs2 - / -Paraffin-embedded samples of brain tissue from mice were cut and 4-micrometer sections were boiled in antigen retrieval solution (DAKO) in a pressure cooker to unmask antigens. Sections were blocked with 3% BSA for 1 h and subsequently incubated with primary antibody against adenylate cyclase III (Santa Cruz Biotechnology) diluted in 3% BSA overnight at 4°C. Alexa Fluor 488 secondary antibody (Invitrogen) was used at a dilution of 1:1000. Images were acquired with a Leica DM5500B microscope fitted with 40x and 60x objectives using Leica Application Suite Advance Fluorescence software (Leica Microsystems).

[0194] Observations and Results Bbs2 of the quinuclidine compounds described herein - / - The effect on metabolic parameters in mice was evaluated. Metabolic abnormalities in BBS patients are known to be one of the main causes of morbidity and contribute to many secondary features of the disease. This example shows that treatment with compound 1 caused a significant decrease in food consumption, body weight, and body fat (Figure 3A) (*p<0.05).

[0195] Serum leptin, a hormone extracted by adipose tissue, was reduced in Bbs2 mice compared to wild-type animals. - / - When treated with Compound 1, serum leptin is reduced to wild-type levels (FIG. 3A).

[0196] Obesity in BBS has been suggested to be associated with two components: a peripheral component and a CNS-related component (see Marion V. et al., Cell Met., 16:363-377 (2012)). To analyze the role of the peripheral component in BBS-associated obesity, we analyzed Bbs2. - / - The effect of treatment on adipogenesis in mice was examined. - / -Analysis of white adipose tissue from mice yielded a heterogeneous population of adipocytes with a significant increase in adipocyte size compared to wild-type controls. Treatment with Compound 1 reduced adipocyte size (Figure 3B) (*p<0.05). - / - The increase in adipocyte size in mice correlated with increased expression of the pro-adipogenic gene Srebf1, which was corrected upon treatment to levels observed in Wt controls (Figure 3C) (*p<0.05).

[0197] To determine whether treatment with the quinuclidine compounds described herein has an effect on metabolic parameters in animals with established disease, Bbs2 - / - Mice were treated with a diet containing 0.033% w / w of Compound 1 for 2 months starting at 4 months of age. As shown in Figure 4A, this treatment reduced body weight, body fat and serum leptin after 1 month of treatment compared to untreated control animals. Analysis of adipose tissue at the final time point after 2 months of treatment also resulted in a reduction in adipocyte size (Figure 4B) (*p<0.05). These data suggest that the treatment may have an effect in a patient population with established disease, e.g., obese patients.

[0198] The CNS component of obesity has been suggested to be associated with loss of cilia in the hypothalamus, resulting in a decrease in leptin signaling and increased food consumption (see Guo, et al., PLOS., DOI:10.1371 (2016)). With this in mind, we also analyzed cilia in the hypothalamus. - / - A loss of cilia was observed in mice. This finding was consistent with the increased food consumption observed in this mouse model (see above). Furthermore, treatment with compound 1 reduced the Bbs2 - / - Hypothalamic cilia were preserved in mice (Figure 5). Taken together, these data suggest that treatment may have effects on both peripheral and CNS components of obesity associated with BBS. EXAMPLES

[0199] Bbs2 - / - Effect of Compound 1 on gene expression in mouse adipose tissue RNA extraction, next-generation sequencing library construction and data analysis Total RNA was extracted using TRIzol (Thermo Fisher Scientific) and chloroform extraction from 4 mice previously aged 6 months, Wt (Wt#1-4), Bbs2 - / - (Bbs#1-4), or Bbs2 treated with 0.033% w / w Compound 1 in the diet (Bbs+Cmpd1#1-4) from 1 to 6 months of age. - / - RNA samples were isolated from adipose tissue that had been sectioned and flash frozen. RNA samples were further purified with an RNAeasyMini Kit (Qiagen) to remove genomic DNA. RNA concentration and purity were assessed using a NanoDrop 8000 microspectrophotometer (Thermo Fisher Scientific). RNA integrity was then assessed using a 4200 TapeStation System (Agilent Technologies).

[0200] Sequencing libraries were generated using the TruSeq Stranded mRNA Library Prep Kit according to the manufacturer's recommendations (Illumina). Sequencing was performed using the High Output NextSeq 500 / 550 v2.5 kit on the Illumina NextSeq 500 platform (2 × 75 bp paired-end reads).

[0201] Data analysis was performed using Array Studio V10.1 (Omicsoft Corporation, a Qiagen company) and mapped to Genome Reference Consortium Mouse Build 38. Heat maps are generated using the Center Scale normalization algorithm on the median normalized Log2 expression values ​​according to the software developer (Omicsoft).

[0202] Discussion and Results Obesity is a major clinical feature of ciliopathies such as BBS (Beales, P., Curr. Opin. Genet. Dev., 15:315-323 (2005)). To better understand the role of compound 1 on adipogenesis, RNA sequencing analysis was performed on Wt, Bbs2 - / - and - Bbs2 treated with compound 1 - / The study was performed on adipose tissue from mice. Wt, Bbs2 - / - and Bbs2 - / - Heatmap analysis of differentially expressed mRNAs in adipose tissue from the +Compound 1 group showed 81 differentially expressed genes in the adipogenic pathway.

[0203] A similar analysis was performed for genes involved in the glycosphingolipid pathway and genes involved in the formation and homeostasis of the primary cilium, also referred to herein as ciliogenesis. Heat maps showed 33 and 52 differentially expressed genes in the glycosphingolipid and cilia pathways, respectively.

[0204] These data suggest that treatment with compound 1 normalizes the expression of genes in dysregulated pathways involved in adipogenesis, ciliogenesis, and genes involved in sphingolipid homeostasis that are dysregulated in adipose tissue associated with BBS. EXAMPLES

[0205] Effect of GCS inhibition on liver abnormalities Measurement of ALT and triglyceride levels Triglyceride and ALT levels were measured using a VetACE™ analyzer (Alfa Wasserman, West Caldwell, NJ).

[0206] Observations and Results Bbs2 - / - The effect of treatment with compound 1 on liver abnormalities in mice was also investigated. BBS patients have been shown to develop a liver phenotype that is strongly associated with obesity (see Day et al., Clin. Genet., 89:507-509 (2015)). Similar to these patients, Bbs2 - / - Mice were characterized as having several liver abnormalities, including elevations in liver weight, serum ALT and triglycerides, which were normalized when treated with Compound 1 ( FIG. 6 ). These data suggest that the effects of treatment with the quinuclidine compounds described herein may also ameliorate hepatic phenotypes associated with obesity. EXAMPLES

[0207] Effect of Compound 1 on retinal degeneration Retinal degeneration Optical coherence tomography (OCT), a non-invasive imaging technique using a Bioptigen Envisu R2200 instrument, was used to generate cross-sectional images of the retina with a resolution of 2 microns to measure the thickness of retinal cell layers in vivo.

[0208] Immunofluorescence Wt and Bbs2 - / - Paraffin-embedded eye samples from mice were prepared and analyzed as described above. Primary antibodies used were rhodopsin (Thermo Scientific) and cone arrestin (EMD Millipore).

[0209] Observations and Results Several mouse models of BBS show progressive retinal degeneration that leads to blindness shortly after birth (see Tobin JL et al., Pediatr. Nephrol., 7:926-936 (2007) and Nishimura DY et al., supra). - / - The mouse model was also characterized by extensive degeneration of the outer nuclear layer (ONL), a reduction in the number of rods and cones, and increased levels of apoptosis in the retina. These changes were found to be progressive, and Bbs2 - / - This was observed at early stages in animals (Figure 7A). Treatment with Compound 1 from 1 to 5 months of age increased the ONL thickness by approximately 2-fold (Figure 7B) (*p<0.05). Improvement of retinal cytoarchitecture, evident by partial restoration / preservation of rod-cone specific staining (Figure 7C), was also observed with treatment with Compound 1. EXAMPLES

[0210] Effects of Compound 1 on the main olfactory epithelium (MOE) Smell Animal olfaction was tested using the protocol adopted by Yang M. et al. (Curr Protoc Neurosci, DOI:10.1002 / 0471142301.ns0824s48(2009)). Prior to testing, animals were acclimated for 3 days and then fasted for 18 hours in Alpha Dri bedding cages. Rewards (Bioserv Supreme Mini-Treats, chocolate flavor) were buried at a depth of 1 cm in clean cages with 3 cm deep bedding. The time when the animal was placed in the cage and found and started to eat the reward was recorded. If the subject was unable to find the buried food after 10 minutes, the test was stopped and the latency score was recorded as 10 minutes.

[0211] Immunofluorescence Wt and Bbs2 - / -Paraffin-embedded samples of the nasal cavity from mice were prepared and analyzed as described above. Primary antibodies used were acetylated tubulin (Cell Signaling Technology), cytokeratin 14 (Protein Tech), SRY-Box 2 (Cell Signaling Technology), doublecortin (Cell Signaling Technology), and olfactory marker protein (Wako).

[0212] Discussion and Results Treatment with the quinuclidine compounds described herein was found to preserve cilia in the main olfactory epithelium (MOE). Improvement in olfaction was assessed in vivo using an olfactory function test based on assessing the latency to find a buried reward. - / - Mice had an olfactory defect compared to Wt animals, which was restored by treatment with Compound 1 (FIG. 8A) (*p<0.05).

[0213] Histological and immunofluorescence analysis revealed that Bbs2 - / - A significant decrease in cilia specific staining (acTubulin) was observed in mouse MOE (Figure 8B). - / - No obvious abnormalities in the respiratory epithelium were observed in mice (data not shown).

[0214] Treatment with compound 1 improved olfaction, which correlated with the preservation / restoration of cilia in the MOE (Figure 8B). Consistent with the in vivo data, Bbs2 - / -The mouse MOE was characterized by a decrease in the amount of adenylate cyclase III (ACIII), an enzyme that initiates the odorant signaling cascade. Treatment increased ACIII, suggesting activation of the odorant signaling cascade (Figure 8C). Further investigations along these same lines also provided the effect of treatment with the quinuclidine compounds described herein on cell differentiation in the MOE. The primary olfactory epithelium is a multicellular layer composed of four distinct cell layers: basal cells (stem cells), supporting cells, immature neurons, and mature neurons, which can be regenerated via differentiation of stem cells into mature neurons (McIntyre J et al., Nat Med, 18:1423-1428 (2012)). This differentiation process is associated with the upregulation of Bbs2, as demonstrated by the accumulation of immature neurons and the decrease of mature neurons. - / - In mice, the phenotype is impaired. Treatment with compound 1 corrected this phenotype to resemble that of wild-type animals (FIG. 9). These data suggest that the mechanism of action of the quinuclidine compounds described herein is via the regulation of differentiation.

[0215] The results shown in the above examples provide the in vivo effects of the quinuclidine compounds described herein on ciliopathies and successfully demonstrate the therapeutic potential of administering the quinuclidine compounds described herein to treat ciliopathies. EXAMPLES

[0216] Effect of compound 1 on adipocyte differentiation In vitro adipocyte differentiation assay To examine the effect of Compound 1 on other mutations, an in vitro differentiation assay in human preadipocytes was developed. The BBS genes BBS1, BBS2, or BBS10 were knocked down in cells using siRNA. For this, human preadipocyte SQ cells (Lonza) were cultured at 30,000 cells / cm in PGM-2 Preadipocyte Growth Medium-2 BulletKit, (Lonza). 2Cells were plated at 100 ng / ml density and grown overnight at 37°C. The next day, cells were transfected with BBS1, BBS2 or BBS10 specific siRNA in OptiMem (Invitrogen) in the presence of Lipofectamine 2000 (Invitrogen) and differentiated for 10 days in RDM-2 medium (Lonza) with or without compound 1 at concentrations of 1.25, 1.5, 5.0 and 10 μM (stock solution of compound 1 was prepared in 100% ethanol and diluted in RDM-2 medium to final concentration before adding to cells). A mixture of non-specific siRNA (scrambled) was used as a negative control. Culture medium was collected for leptin analysis using Quantikine ELISA kit (RnD Systems, Inc.) at days 0, 5, 7 and 10 of adipocyte differentiation. Photomicrography (Axiovert 25, Zeiss with LASv4.2, Leica) was used to quantify lipid accumulation, evident by the size and amount of lipid vacuoles in the cells.

[0217] Discussion and Results The effect of the quinuclidine compounds described herein on adipocyte differentiation was evaluated as measured by lipid accumulation and leptin secretion.The effect of BBS gene knockdown on adipocyte differentiation is evident by the increase in lipid accumulation in cells (Figure 10A) and the increase in the secretion of leptin into the medium (Figure 10B).It is shown that adipogenesis is suppressed by treatment with compound 1, resulting in a decrease in lipid accumulation (Figure 10C) and leptin secretion (Figure 10D) in a dose-dependent manner. EXAMPLES

[0218] In-vitro GCS inhibition (compound 1 and analogues) The 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. A specific protocol is provided below.

[0219] Microsomal assay for glucosylceramide synthase activity: Enzyme assay using microsomes as source of glucosylceramide synthase activity. Fluorescent ceramide substrate is delivered to the membrane-bound enzyme as a complex with albumin. After reaction, ceramide and glucosylceramide are separated and quantified by reversed-phase HPLC with fluorescence detection. Enzyme activity is assessed using a fluorescently labeled substrate and microsomes as source of glucosylceramide synthase. C6-NBD-ceramide is delivered in 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) is achieved by reversed-phase HPLC with fluorescence detection.

[0220] 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 min at 4° C. The supernatant is removed and clarified by additional centrifugation at 100,000 g for 1 h at 4° C. The pellet is then resuspended in lysis buffer, aliquoted, and stored at −80° C. before use.

[0221] Glucosylceramide synthase assay To determine glucosylceramide synthase inhibition, substrates (fluorescent ceramide and UDP-glucose, 3 μM and 4 μM, respectively) at twice their Km are combined 1:1 with microsomes (diluted 1:50) and incubated for 1 h at room temperature on a plate shaker in the dark. The reaction is stopped by adding 150 μL of 100 μM C8-ceramide in 50% aqueous isopropanol; 10 μL of the final mix is ​​analyzed by HPLC (equipped with a fluorescence detector). The mobile phase is 1% formic acid in 81% methanol / 19% water, with a flow rate of 0.5 mL / min. Fluorescence is measured at λ ex = 470 nm and λ em = 530 nm. Under these conditions, the retention time of NBD-C6-GluCer is about 1.7 min, and NBD-C6-Cer elutes from the column after about 2.1 min. Both peaks were separated from each other and from the baseline and were automatically integrated by the HPLC software. The conversion rate of substrate to product is used as the readout for the inhibitor test.

[0222] GM3 Fluorescence Linked Immunosorbent Assay (FLISA): This is a phenotypic assay that measures GM3 expression in B16 mouse melanoma or C32 human melanoma cells following treatment with a test compound. Cell surface GM3 expression is determined by antibody-mediated fluorescence.

[0223] Compounds are diluted in 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. Plates are incubated for 48 hours at 37°C with 5% CO2, and then washed once with TBS. Anti-GM3 antibody is added to each well, and the plate is then incubated for an additional hour at room temperature. The plate is then washed twice and incubated with labeled secondary antibody for an additional hour. After the final incubation, the plate is washed twice and incubated with λ ex = D640 / 20nm and λ em Fluorescence at 657 nm is detected in a fluorescence reader.

[0224] 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 listed as "GM3B16IC 50 " or "GM3C32IC 50 These values ​​represent the concentration of compound that results in 50% inhibition of GM3 expression at the cell surface.

[0225] [Table 2]

[0226] These comparative results demonstrate that compounds according to the present disclosure have comparable in-vitro activity as GCS inhibitors, and are therefore expected to demonstrate similar in-vivo benefits. EXAMPLES

[0227] 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.

[0228] 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 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.

[0229] Fifty-five 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 in part 1 of the study. Eight healthy men participated in part 2.

[0230] In part 1, subjects were randomized to receive 2, 5, 15, 25, 50, 100, or 150 mg of Compound 2 (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 given either on an empty stomach (at least 10 hours before and 4 hours after dosing) or simultaneously 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.

[0231] In Study 1, Part 1, blood was sampled for plasma concentrations of Compound 2 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 after dosing. Urine samples were collected and analyzed for Compound 2 concentrations beginning 2 hours before and continuing through 48 hours after study drug administration.

[0232] 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.

[0233] From Part 1, the maximum plasma concentrations (C 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 over the entire dose range: a 75-fold dose increase resulted in a geometric mean C max , AUC last , and AUC inf The PK results were 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; t max =C max time to saturation; Vss / F = apparent volume of distribution at steady state):

[0234] [Table 3]

[0235] Part 2 showed that a 5 mg dose administered with a high-fat meal had no effect on exposure to Compound 2 compared to fasting conditions. Median t max was 6.00 h whether fed or fasted. The geometric mean ratio of fed / fasted was C max and AUC last and 0.92 and 0.91, respectively. Within-subject variability (i.e., fed vs. fasted) accounted for less than half of the total subject variability.

[0236] 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.

[0237] The study enrolled and randomized 36 healthy adults (19 men and 17 women) (n=9 per group) to receive Compound 2 at 5, 10, or 20 mg (provided in the form of a 5-mg capsule of the L-malate salt) or placebo, administered once daily after at least a 10-hour fast, for 14 days.

[0238] Blood was sampled for plasma concentrations of Compound 2 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; 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 consecutively on Day 14. Pharmacodynamic endpoints (plasma GL-1, GL-3, and GM3 concentrations) were assessed 0 hours post-dose on Days 1-5, 8, 11, 13, and 14; and 24 hours post-dose on Day 15.

[0239] In subjects receiving 5, 10, or 20 mg of Compound 2 once daily for 14 days, plasma C max were observed to occur 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 dose range of 5 to 20 mg: this four-fold dose increase resulted in a geometric mean C 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 following table:

[0240] [Table 4]

[0241] 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%, without any clear dose-related association. R(0-24) ranged between 1.49 L / h and 2.07 L / h, approximately 3.18-3.86-fold lower than the observed plasma CL / F.

[0242] 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):

[0243] [Table 5]

[0244] 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.

[0245] 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, respectively, in the 10 and 20 mg dose groups.

[0246] 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.

[0247] C of Compound 2 in the 5, 10, and 20 mg dose groups trough The mean estimated reductions from baseline in plasma GL-1 attributable to treatment (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.

[0248] conclusion In these studies, exposure to compound 2 (C maxand AUC) were approximately dose proportional when administered over a range of 2 to 150 mg as a single dose or 5 to 20 mg as repeated daily doses for 14 days. A high-fat meal had no effect on exposure in subjects receiving a single 5 mg dose compared with fasting. At repeated daily doses of 5 to 20 mg, steady state was reached within 5 days; neither age nor sex influenced accumulation. Pharmacodynamically, repeated 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.

[0249] 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 any study.

[0250] No clinically relevant hematological 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 the 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.

[0251] While the 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 invention pertains.

[0252] Furthermore, when features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also described with respect to any individual members or subgroups of members of the Markush group.

[0253] 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 the case of conflict, the present specification, including definitions, will control.

Claims

1. (a) for treating a ciliopathies selected from Joubert syndrome, Meckel-Gruber syndrome, Senior-Loken syndrome, Orofacial-Digital syndrome type I, Leber congenital amaurosis, Bardet-Biedl syndrome (BBS), Amström syndrome, June asphyxiating thoracic dystrophy, Ellis-van Creveld syndrome, Sensenbrenner syndrome and primary ciliary dyskinesia in a subject in need thereof; or (b) for treating a disease or disorder selected from obesity, liver disease, retinal degeneration, olfactory dysfunction, hyperlipidemia, type 2 diabetes mellitus, and metabolic syndrome in a subject suffering from a ciliopathy selected from Joubert syndrome, Meckel-Gruber syndrome, Senior-Loken syndrome, Orofacial-Digital syndrome type I, Leber congenital amaurosis, Bardet-Biedl syndrome (BBS), Amström syndrome, June asphyxiating thoracic dystrophy, Ellis-van Creveld syndrome, Sensenbrenner syndrome, and primary ciliary dyskinesia; A pharmaceutical composition for use in the method, comprising an effective amount of a compound represented by formula (IX) or (XI) 【Chemistry 1】 (In the formula: R 4 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 -alkyloxy) or a pharma- ceutically acceptable salt thereof, The pharmaceutical composition.

2. R 4 The pharmaceutical composition of claim 1 , wherein is fluorine.

3. 2. The pharmaceutical composition of claim 1, wherein the compound is 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; (S)-quinuclidin-3-yl (2-(4'-(2-methoxyethoxy)-[1,1'-biphenyl]-4-yl)propan-2-yl)carbamate; and pharma-ceutically acceptable salts thereof.

4. The pharmaceutical composition of any one of claims 1 to 3, 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.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the ciliopathic disease is BBS.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the subject is a mammal, such as a human.

7. The pharmaceutical composition can be administered by systemic administration, e.g., by a non-parenteral route. The pharmaceutical composition according to any one of claims 1 to 6, wherein the pharmaceutical composition is administered via the intravenous route.

Images