WO2014011863A1

WO2014011863A1 - Method of treatment with 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane

Info

Publication number: WO2014011863A1
Application number: PCT/US2013/050057
Authority: WO
Inventors: Kristen G. Jordan; Sharon Rae Letchworth
Original assignee: Targacept Inc
Current assignee: Gyre Therapeutics Inc
Priority date: 2012-07-12
Filing date: 2013-07-11
Publication date: 2014-01-16
Anticipated expiration: 2015-01-12

Description

METHOD OF TREATMENT WITH

3-CYCLOPROPYLCARBONYL-3,6-DIAZABICYCLO[3.1.1]HEPTANE

Field of the Invention

The present invention relates to 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1]heptane, its salt forms, pharmaceutical compositions of these salt forms, and methods for treating a wide variety of conditions and disorders.

Background of the Invention

The compound, 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane (Compound A), is a neuronal nicotinic receptor (NNR) agonist with selectivity for α4β2* (a432-containing) and α6β2^* (a632-containing) NNRs. Compound A demonstrates efficacy in, among other things, preventing full onset of abnormal involuntary movements (AIMs) and also in attenuating existing AIMs in preclinical rodent models of levodopa-induced dyskinesias (LIDs). Compound A reduces LIDs in non-human primates (macaques) and does not impede general activity or levodopa's (L-dopa's) effect on motor deficits. Compound A exhibits neuroprotective effects against MPP+ toxicity in primary cultures of rat dopamine neurons. When tested in a nonhuman primate model, Compound A significantly reduced LIDs. This effect was especially strong at a 0.10 mg/kg dose. Compound A did not significantly alter either total on-time (total time when L-dopa's anti-parkinsonian effects are observed) or parkinsonian disability (the effectiveness of L-dopa treatment). Compound A significantly decreased the percent of bad quality on-time (time characterized by troubling dyskinesia) from 50-55% to approximately 30% and correspondingly increased the percent of good quality on-time (time without troubling dyskinesia) from 45-50% to approximately 70%. Total dyskinesias, as well as chorea and dystonia endpoints, were significantly decreased. Compound A was very well tolerated in the study, did not negatively impact the general motor activity of the animals, and did not counteract levodopa's effect on parkinsonism.

Compound A has the following structural formula:

One synthesis of Compound A is disclosed in PCT Application No.

PCT/US2010/058836 (WO 201 1/071758), which is incorporated by reference. Another synthesis, including formation of preferred salts, is disclosed in PCT/US2012/028691 , which is incorporated by reference.

Summary of the Invention One aspect of the present invention is a method, use, compound for use, or use for preparation of a medicament for treating or preventing one or more disease or disorder. In one embodiment, the disease or disorder is one or more tremor, including but not limited to Essential tremor, Parkinsonian tremor, Cerebellar tremor (including intention tremor and titubation), Rubral tremor (Holmes' tremor), Dystonic tremor, Wilson's disease, Tremor in multiple sclerosis, Tremor resulting from a neurodegenerative disorder other than those specifically listed, Tremor resulting from basal ganglia disease other than Parkinson's disease and dystonia, Familial tremor other than essential tremor and Wilson's disease, Tremor resulting from peripheral neuropathy, Tremor resulting from stroke, Tremor resulting from traumatic brain injury, Tumor-induced tremor, Enhanced or exaggerated physiologic tremor, Tremor resulting from metabolic disorders, Tremor resulting from chronic kidney disease, Tremor resulting from liver failure, Tremor resulting from phenylketonuria, Orthostatic tremor, Psychogenic tremor, Drug-induced tremor, Tremor resulting from alcoholism (asterixis), and Tremor resulting from toxins (e.g. heavy metal poisoning). In another embodiment, the disease or disorder is one or more dystonia, including but not limited to, Genetic or familial dystonia (one or more genes associated with various forms of dystonia), Myoclonic dystonia, General dystonia, lodiopathic torsion dystonia, Dystonia associated with Parkinson's disease, Dystonia associated with Wilson's disease, Focal dystonia such as, blepharospasm, cervical dystonia or torticollis, cranial dystonia, oromandibular dystonia, or spasmodic dystonia, Multifocal dystonia, Cranial dystonia or Meige's syndrome, Segmental dystonia, Hemidystonia, Torsion dystonia, Paroxysmal dystonia, Dystonia resulting from trauma, Dystonia resulting from stroke, Tumor-induced dystonia, Dystonia resulting from infection, Dystonia resulting from oxygen deprivation, Dystonia resulting from toxins (e.g. heavy metal poisoning, carbon dioxide poisoning), and Drug-induced dystonia.

Other aspects and embodiments of the present invention will be described herein. The scope of the present invention includes combinations of aspects, embodiments, and preferences.

Brief Description of the Figures

Figure 1 illustrates graphically the effects of acute propranolol and sub-chronic

Compound A administration on harmaline-induced tremor events. Mice (n = 12/group) were treated with Compound A (0.01 , 0.1 , or 1 mg/kg; i.p.) BID for seven (7) days and again 20 minutes prior to harmaline (30 mg/kg; s.c.) on test day (Day 8). Tremor events were measured 10 minutes following harmaline injection. Data are presented as mean ± SEM.

Detailed Description

Definitions The following definitions are meant to clarify, but not limit, the terms defined. If a particular term used herein is not specifically defined, such term should not be considered indefinite. Rather, terms are used within their accepted meanings.

3-Cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane may also be referred to as 3,6- diazabicyclo[3.1.1]heptan-3-yl(cyclopropyl)methanone, or, potentially, still other chemical names, depending upon the naming convention used. The choice of naming convention should not affect the scope of the present invention. As noted herein, the structure of the compound is:

and, for ease of reference, may also be referred to herein as Compound A.

The phrase "compound of the present invention" as used herein refers to 3- cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane or an acid addition salt thereof. The acid preferably is selected from hydrochloric, p-toluenesulfonic, L-aspartic, maleic, L-glutamic, 1- hydroxy-2-naphthoic (namely, xinafoate), fumaric, galactaric, hippuric, L-mandelic, succinic, adipic, or (+)-camphoric. In preferred embodiments, the acid addition salt is a p- toluenesulfonate, maleate, galactarate, benzoate, hippurate, xinafoate, or (+)-camphorate. In still more preferred embodiments, the salt is a galactarate, benzoate, hippurate, or xinafoate. A most preferred salt form is a hemigalactarate. The phrase "compound of the present invention" includes a hydrated or a solvated salt form.

Further, as used herein, the term "compound" may be used to mean the free base form, or alternatively, a salt form of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane, depending on the context, which will be readily apparent.

As used herein, the term "pharmaceutically acceptable" refers to carrier(s), diluent(s), excipient(s) or salt forms that are compatible with the other ingredients of the formulation and not deleterious to the recipient of the pharmaceutical composition.

As used herein, the term "pharmaceutical composition" refers to a compound of the present invention optionally admixed with one or more pharmaceutically acceptable carriers, diluents, excipients, or adjuvants. Pharmaceutical compositions preferably exhibit a degree of stability to environmental conditions so as to make them suitable for

manufacturing and commercialization purposes.

As used herein, the terms "effective amount," "therapeutic amount," or "effective dose" refer to an amount of active ingredient sufficient to elicit the desired pharmacological or therapeutic effects, thus resulting in effective prevention or treatment of a disorder. Prevention of a disorder may be manifested by delaying or preventing the progression of the disorder, as well as delaying or preventing the onset of the symptoms associated with the disorder. Treatment of the disorder may be manifested by a decrease or elimination of symptoms, inhibition or reversal of the progression of the disorder, as well as any other contribution to the well being of the patient.

The effective dose can vary, depending upon factors such as the condition of the patient, the severity of the symptoms of the disorder, and the manner in which the pharmaceutical composition is administered. Typically, to be administered in an effective dose, compounds are required to be administered in an amount of less than 5 mg/kg of patient weight. Often, the compounds may be administered in an amount from less than about 1 mg/kg patient weight to less than about 100 μg/kg of patient weight, and

occasionally between about 10 μg/kg to less than about 100 μg/kg of patient weight. The foregoing effective doses typically represent that amount administered as a single dose, or as one or more doses administered over a 24 h period. For human patients, the effective dose of the compounds may require administering the compound in an amount of at least about 1 mg/24 hr/patient, but not more than about 1000 mg/24 hr/patient, and often not more than about 500 mg/ 24 hr/ patient. Potential doses may be in the range of 500 μg to 2 mg, as free base equivalents.

As used herein, the phrase "substantially crystalline" includes greater than 20%, preferably greater than 30%, and more preferably greater than 40% (e.g. greater than any of 50, 60, 70, 80, or 90%) crystalline.

The term "stability" as defined herein includes chemical stability and solid state stability, where the phrase "chemical stability" includes the potential to store salts of the invention in an isolated form, or in the form of a formulation in which it is provided in admixture with pharmaceutically acceptable carriers, diluents, excipients, or adjuvants, such as in an oral dosage form, such as a tablet, capsule, or the like, under normal storage conditions, with an insignificant degree of chemical degradation or decomposition, and the phrase "solid state stability", includes the potential to store salts of the invention in an isolated solid form, or in the form of a solid formulation in which it is provided in admixture with pharmaceutically acceptable carriers, diluents, excipients, or adjuvants, such as in an oral dosage form, such as a tablet, capsule, or the like, under normal storage conditions, with an insignificant degree of solid state transformation, such as crystallization, recrystallization, solid state phase transition, hydration, dehydration, solvation, or desolvation.

Examples of "normal storage conditions" include one or more of temperatures of between -80 °C and 50 °C, preferably between 0 °C and 40 °C and more preferably ambient temperatures, such as 15 °C to 30 °C, pressures of between 0.1 and 2 bars, preferably at atmospheric pressure, relative humidity of between 5 and 95%, preferably 10 to 60%, and exposure to 460 lux or less of UV/visible light, for prolonged periods, such as greater than or equal to six months. Under such conditions, salts of the invention may be found to be less than 5%, more preferably less than 2%, and especially less than 1 %, chemically degraded or decomposed, or solid state transformed, as appropriate. The skilled person will appreciate that the above-mentioned upper and lower limits for temperature, pressure, and relative humidity represent extremes of normal storage conditions, and that certain combinations of these extremes will not be experienced during normal storage (e.g., a temperature of 50 °C and a pressure of 0.1 bar).

Compounds

One embodiment of the present invention includes 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1]heptane (Formula I) or a pharmaceutically acceptable salt thereof.

Formula I

Compound A

One embodiment includes use of 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1]heptane or a pharmaceutically acceptable salt thereof in the manufacture of a medicament.

One embodiment of the present invention includes a method, use, compound for use, or use for preparation of a medicament for treating or preventing of a variety of diseases, disorders, or dysfunctions, comprising administering to a mammal in need of such treatment, a therapeutically effective amount of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane or a pharmaceutically acceptable salt thereof. In one embodiment, the disease, disorder, or dysfunction is one or more tremor, including but not limited to Essential tremor, Parkinsonian tremor, Cerebellar tremor (including intention tremor and titubation), Rubral tremor (Holmes' tremor), Dystonic tremor, Wilson's disease, Tremor in multiple sclerosis, Tremor resulting from a neurodegenerative disorder other than those specifically listed, Tremor resulting from basal ganglia disease other than Parkinson's disease and dystonia, Familial tremor other than essential tremor and Wilson's disease, Tremor resulting from peripheral neuropathy, Tremor resulting from stroke, Tremor resulting from traumatic brain injury, Tumor-induced tremor, Enhanced or exaggerated physiologic tremor, Tremor resulting from metabolic disorders, Tremor resulting from chronic kidney disease, Tremor resulting from liver failure, Tremor resulting from phenylketonuria, Orthostatic tremor, Psychogenic tremor, Drug- induced tremor, Tremor resulting from alcoholism (asterixis), and Tremor resulting from toxins (e.g. heavy metal poisoning). In another embodiment, the disease, disorder, or dysfunction is one or more dystonia, including but not limited to, Genetic or familial dystonia (one or more genes associated with various forms of dystonia), Myoclonic dystonia, General dystonia, lodiopathic torsion dystonia, Dystonia associated with Parkinson's disease,

Dystonia associated with Wilson's disease, Focal dystonia such as, blepharospasm, cervical dystonia or torticollis, cranial dystonia, oromandibular dystonia, or spasmodic dystonia, Multifocal dystonia, Cranial dystonia or Meige's syndrome, Segmental dystonia,

Hemidystonia, Torsion dystonia, Paroxysmal dystonia, Dystonia resulting from trauma, Dystonia resulting from stroke, Tumor-induced dystonia, Dystonia resulting from infection, Dystonia resulting from oxygen deprivation, Dystonia resulting from toxins (e.g. heavy metal poisoning, carbon dioxide poisoning), and Drug-induced dystonia.

One embodiment of the present invention includes a pharmaceutical composition comprising a therapeutically effective amount of 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1]heptane or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carrier. Another embodiment of the present invention includes the use of a pharmaceutical composition of the present invention in the manufacture of a medicament for treatment of central nervous system disorders and dysfunctions. Another embodiment of the present invention includes 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1 Jheptane or a pharmaceutically acceptable salt thereof with reference to any one of the Examples. Another embodiment of the present invention includes 3- cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane or a pharmaceutically acceptable salt thereof for use as an active therapeutic substance.

Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structure except for the replacement of a hydrogen atom by deuterium or tritium, or the replacement of a carbon atom by ¹³C or ¹⁴C, or the replacement of a nitrogen atom by ¹⁵N, or the replacement of an oxygen atom with ¹⁷0 or¹⁸0 are within the scope of the invention. Such isotopically labeled compounds are useful as research or diagnostic tools.

As noted herein, the present invention includes specific representative compounds, which are identified herein with particularity. The compounds of this invention may be made by a variety of methods, including well-known standard synthetic methods. Illustrative general synthetic methods are set out below and then specific compounds of the invention are prepared in the working Examples.

In all of the examples described below, protecting groups for sensitive or reactive groups are employed where necessary in accordance with general principles of synthetic chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T. W. Green and P. G. M. Wuts, Protecting Groups in Organic Synthesis, 3^rd Edition, John Wiley & Sons, New York (1999)). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art. The selection of processes as well as the reaction conditions and order of their execution shall be consistent with the preparation of compounds of the present invention.

The present invention also provides a method for the synthesis of compounds useful as intermediates.

General Synthetic Methods

The manner in which Compound A can be synthesized can vary. In one approach,

Compound A can be prepared via the coupling of a 3,6-diazabicyclo[3.1.1]heptane in which the 6-position nitrogen atom has been protected (to prevent reaction with acylating agents) with a suitable cyclopropylcarbonyl derivative (such as cyclopropylmethanoyl chloride), followed by removal of the protecting group (typically with acid). Cyclopropylmethanoyl chloride may be prepared by treatment of the cyclopropylcarboxylic acid with, among other reagents, thionyl chloride or oxalyl chloride.

The synthesis of a suitably protected 3,6-diazabicyclo[3.1.1]heptane, 6-(tert- butoxycarbonyl)-3,6-diazabicyclo[3.1.1]heptane, is disclosed in WO 2005/108402 to Pinna, et al. (incorporated by reference with regard to such synthetic teaching). Those skilled in the art of organic synthesis will recognize that other suitably protected 3,6- diazabicyclo[3.1.1]heptanes can also be used to prepare Compound A (see, for example, T. W. Greene and P.G. M. Wuts, Protective Groups in Organic Synthesis, 3^rd Edition, John Wiley & Sons, New York (1999).

Another means of making Compound A is to couple a suitable 6-protected-3,6- diazabicyclo[3.1.1]heptane with a cyclopropylcarboxylic acid, followed by removal of the protecting group. This approach typically requires the use of a suitable activating agent, such as Ν,Ν'-dicyclohexylcarbodiimide (DCC), (benzotriazol-1- yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazol-1- yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), O-(benzotriazol-l-yl)- N,N,N',N'-bis(tetramethylene)uronium hexafluorophosphate (HBPyU), O-(benzotriazol-l-yl)- Ν,Ν,Ν',Ν'-tetramethyluronium hexafluorophosphate (HBTU), 0-(benzotriazol-1-yl)-N,N,N',N'- tetramethyluronium tetrafluoroborate (TBTU), or (1-ethyl-3-(3- dimethylaminopropyl)carbodiimide) (EDCI) with 1-hydroxybenzotriazole (HOBt). Other activating agents are well known to those skilled in the art, for example, see Kiso and Yajima, Peptides, pp 39-91 , Academic Press, San Diego, CA (1995). Scheme 1 : Synthesis of Compound A

Bn = benzyl; Ms = methanesulfonyl; DMF = dimethylformamide; THF = tetrahydrofuran;

TEA = triethylamine; DCM = dichloromethane; NMP = N-methyl-2-pyrrolidone.

Alternatively, and advantageously, Compound A can be made using a reaction sequence similar to that shown in Scheme 1 . This approach involves a ring closing process in which the anion of cyclopropylcarboxamide is reacted with a bis-electrophile, such as intermediate 6, decreasing the overall length of the synthesis compared with previously described syntheses (for instance, WO 201 1/071758). A specific example of this approach to the synthesis of Compound A is given in the Examples Section and summarized in Scheme 1 . However, the reagents used to accomplish the transformations of this approach can vary. For instance, a variety of alcohols can be used the esterification reaction, providing a variety of diester products (an example being diester 3). The physical properties of particular diesters may provide advantages in purification or handling. Likewise, a variety of amines and solvents can be utilized in the ring closure of the 4-membered (azetidine) ring, a particular amine/solvent mixture providing advantages in either reactivity or product purity (including stereochemical purity). The reduction of the dialkyl azetidinyl-2,4-dicarboxylate (intermediate 4, for example) to the corresponding dialcohol (such as dialcohol 5) can be accomplished by a variety of reagents (e.g., borohydride and aluminumhydride reagents) in a variety of solvents. It is important to choose a reducing reagent that preserves the cis relative stereochemistry around the azetidine ring. Conversion of the dialcohol into a suitable bis-electrophile can be accomplished by a variety of reagents known to those of skill in the art. Reactions for converting alcohols to the corresponding halides (e.g., chlorides, bromides, iodides), as well as those for converting alcohols to sulfonates, phosphates, and the like, are all well known in the chemical literature. The closure of the 6-membered (piperidine) ring with cyclopropylcarboxamide can also be accomplished with a variety of reagents, including a variety of bases for production of the anion and a variety of solvents. Finally, the deprotection of the azetidinyl nitrogen can be accomplished by a variety of conditions, depending on the nature of the protecting group.

Those skilled in the art of organic synthesis will appreciate that there are multiple means of producing Compound A in which one or more atoms are labeled with a

radioisotope appropriate to various uses. For example, coupling of ¹¹C-labeled

cyclopropylcarboxylic acid with 6-(tert-butoxycarbonyl)-3,6-diazabicyclo[3.1.1]heptane, using one of the activating agents listed above, will produce 3-¹¹C-labeled-cyclopropylcarbonyl-6- (tert-butoxycarbonyl)-3,6-diazabicyclo[3.1.1 Jheptanes. Subsequent removal of the tert- butoxycarbonyl protecting group, as described above will produce a compound suitable for use in positron emission tomography. Likewise, coupling of ³H- or ¹⁴C-labeled

cyclopropylcarboxylic acid with 6-(tert-butoxycarbonyl)-3,6-diazabicyclo[3.1.1]heptanes, followed by removal of the protecting group as described above will produce an isotopically modified Compound A, suitable for use in receptor binding and metabolism studies or as an alternative therapeutic compound.

As mentioned hereinabove, solid salt forms are generally preferred for oral formulations due to their tendency to exhibit these properties in a preferential way; and in the case of basic drugs, such as 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane, acid addition salts are often the preferred salt form.

Different salt forms vary greatly in their ability to impart these properties, and such properties cannot be predicted with accuracy. For example, some salts are solids at ambient temperatures, while other salts are liquids, viscous oils, or gums at ambient temperatures. Furthermore, some salt forms are stable to heat and light under extreme conditions and others readily decompose under much milder conditions. Thus, the development of a suitable acid addition salt form of a basic drug for use in a pharmaceutical composition is a highly unpredictable process. There is a need for salt forms that display improved properties, including purity, stability, solubility, and bioavailability. Preferential characteristics of these novel salt forms include those that would increase the ease or efficiency of manufacture of the active ingredient and its formulation into a commercial product. Lastly, there is a need for stable polymorphic forms of these salts that allows for an increased ease or efficiency of manufacture of the active ingredient and its formulation into a commercially product.

The degree (%) of crystallinity may be determined by the skilled person using x-ray powder diffraction (XRPD). Other techniques, such as solid state NMR, FT-IR, Raman spec- troscopy, differential scanning calorimetry (DSC) and microcalorimetry, may also be used. For compounds of the current invention, it has been found to be possible to produce salts in forms which are substantially crystalline. Several of these crystalline salts demonstrated stability that is sufficient to establish their promise in the production of pharmaceutical preparations. Such stability can be demonstrated in a variety of ways. Propensity to gain and release atmospheric moisture can be assessed by dynamic vapor sorption (DVS). Stability to elevated temperatures and humidity can be studied by storing the solid salts at 40°C/75% RH for up to eight days, and then re-examining each by weight, appearance under the microscope, and XRPD.

Polymorphs

The compounds of the present invention may crystallize in more than one form, a characteristic known as polymorphism, and such polymorphic forms ("polymorphs") are within the scope of the present invention. Polymorphism generally can occur as a response to changes in temperature, pressure, or both. Polymorphism can also result from variations in the crystallization process. Polymorphs can be distinguished by various physical characteristics known in the art such as XRPD patterns (diffractograms), solubility in various solvents, and melting point.

The present invention includes various polymorphic forms of the salt forms of 3- cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane, including hydrates and solvates of the salts. Such polymorphic forms are characterized by their x-ray powder diffraction (XRPD) patterns (diffractograms).

As noted, the salt forms of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane may exist in solvated, for example hydrated, as well as unsolvated forms. The present invention encompasses all such forms.

The present invention also includes isotopically labeled compounds wherein one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, and oxygen, such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸0, and ¹⁷0. Such isotopically labelel compounds are useful as research or diagnostic tools.

Pharmaceutical Compositions

Although it is possible to administer the compounds of the present invention in the form of a bulk active chemical, it is preferred to administer the compound in the form of a pharmaceutical composition or formulation. Thus, one aspect the present invention includes pharmaceutical compositions comprising the compound of the present invention and one or more pharmaceutically acceptable carriers, diluents, or excipients. Another aspect of the invention provides a process for the preparation of a pharmaceutical composition, including admixing the compound of the present invention with one or more pharmaceutically acceptable carriers, diluents or excipients.

The manner in which the compounds of the present invention are administered can vary. The compounds of the present invention are preferably administered orally. Preferred pharmaceutical compositions for oral administration include tablets, capsules, caplets, syrups, solutions, and suspensions. The pharmaceutical compositions of the present invention may be provided in modified release dosage forms such as time-release tablet and capsule formulations.

The pharmaceutical compositions can also be administered via injection, namely, intravenously, intramuscularly, subcutaneously, intraperitoneally, intraarterially, intrathecally, and intracerebroventricularly. Intravenous administration is a preferred method of injection. Suitable carriers for injection are well known to those of skill in the art and include 5% dextrose solutions, saline, and phosphate buffered saline.

The formulations may also be administered using other means, for example, rectal administration. Formulations useful for rectal administration, such as suppositories, are well known to those of skill in the art. The compounds can also be administered by inhalation, for example, in the form of an aerosol; topically, such as, in lotion form; transdermal^, such as, using a transdermal patch (for example, by using technology that is commercially available from Novartis and Alza Corporation), by powder injection, or by buccal, sublingual, or intranasal absorption.

Pharmaceutical compositions may be formulated in unit dose form, or in multiple or subunit doses

The administration of the pharmaceutical compositions described herein can be intermittent, or at a gradual, continuous, constant or controlled rate. The pharmaceutical compositions may be administered to a warm-blooded animal, for example, a mammal such as a mouse, rat, cat, rabbit, dog, pig, cow, or monkey; but advantageously is administered to a human being. In addition, the time of day and the number of times per day that the pharmaceutical composition is administered can vary.

The compounds of the present invention may be used in the treatment of a variety of disorders and conditions and, as such, may be used in combination with a variety of other suitable therapeutic agents useful in the treatment or prophylaxis of those disorders or conditions. Thus, one embodiment of the present invention includes the administration of the compound of the present invention in combination with other therapeutic compounds. For example, the compound of the present invention can be used in combination with other NNR ligands (such as varenicline), antioxidants (such as free radical scavenging agents), antibacterial agents (such as penicillin antibiotics), antiviral agents (such as nucleoside analogs, like zidovudine and acyclovir), anticoagulants (such as warfarin), anti-inflammatory agents (such as NSAI Ds), anti-pyretics, analgesics, anesthetics (such as used in surgery), acetylcholinesterase inhibitors (such as donepezil and galantamine), antipsychotics (such as haloperidol, clozapine, olanzapine, and quetiapine), immuno-suppressants (such as cyclosporin and methotrexate), neuroprotective agents (such as A₂A inhibitors and caffeine), blood-brain barrier permeability enhancers, steroids (such as steroid hormones), corticosteroids (such as dexamethasone, predisone, and hydrocortisone), vitamins, minerals, nutraceuticals, anti-depressants (such as imipramine, fluoxetine, paroxetine, escitalopram, sertraline, venlafaxine, and duloxetine), anxiolytics (such as alprazolam and buspirone), anticonvulsants (such as phenytoin and gabapentin), vasodilators (such as prazosin and sildenafil), mood stabilizers (such as valproate and aripiprazole), anti-cancer drugs (such as anti-proliferatives), antihypertensive agents (such as atenolol, clonidine, amlopidine, verapamil, and olmesartan), laxatives, stool softeners, diuretics (such as furosemide), anti-spasmotics (such as dicyclomine), anti-dyskinetic agents, and anti-ulcer medications (such as esomeprazole). One preferred use of the compounds of the present invention is the treatment and prevention of Parkinson's Disease, AIMs, and LIDs, and thus compounds of the present invention may be used in combination with pharmaceutical agents used to treat Parkinson's Disease, AIMs, and LIDs. Such agents include NNR agonists (α4β2, α7, etc.), dopamine precursors (such as levodopa-carbidopa, levodopa-benserazide, and duodopa), dopamine agonists (such as bromocriptine, cabergoline, lisuride, pergolide, pramipexole, popinirole, talipexole, rotigotine, and apomorphine), dopa carboxylase inhibitors, MAO-B inhibitors (such as selegiline, rasagoline, and safinamide), COMT inhibitors (such as entacapone and tolcapone), antiglutamatergic agents (such as amantadine), anticholinergic agents (such as trihexyphenidyl, benztropine, and biperiden), anti-dementia agents (such as rivastigmine, donepezil, galantamine, and memantine), antipsychotic agents (such as quetiapine, clozapine, and resperidone), antiepileptic agents (such as zonisamide), noradrenaline precursors (such as droxidopa), mGluR4 agonists, mGluR5 inhibitors, 5HT1A 1 B agonists, 5HT2A antagonists/inverse agonists, opioid antagonists (mu, delta, kappa), and AMPA receptor blockers. Such a combination of pharmaceutically active agents may be administered together or separately and, when administered separately, administration may occur simultaneously or sequentially, in any order. The amounts of the compounds or agents and the relative timings of administration will be selected in order to achieve the desired therapeutic effect. The administration in combination of a compound of the present invention with other treatment agents may be in combination by administration concomitantly in: (1 ) a unitary pharmaceutical composition including both compounds; or (2) separate pharmaceutical compositions each including one of the compounds. Alternatively, the combination may be administered separately in a sequential manner wherein one treatment agent is administered first and the other second. Such sequential administration may be close in time or remote in time.

Method of Treatment

3-Cyclopropylcarbonyl-3,6-diazabicyclo[3.1 .1 ]heptane, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing such, can be used for the prevention or treatment of various conditions or disorders for which other types of nicotinic compounds have been proposed or are shown to be useful as therapeutics, such as CNS disorders (including neurodegenerative disorders), inflammation, inflammatory response associated with bacterial and/or viral infection, pain, diabetes, metabolic syndrome, autoimmune disorders, dermatological conditions, addictions, obesity or other disorders described in further detail herein. This compound can also be used as a diagnostic agent in receptor binding studies (in vitro and in vivo). Such therapeutic and other teachings are described, for example, in references previously listed herein, including Williams et al., Drug News Perspec. 7(4): 205 (1994), Arneric et al., CNS Drug Rev. 1(1): 1- 26 (1995), Arneric et al., Exp. Opin. Invest. Drugs 5(1): 79-100 (1996), Yang et al., Acta Pharmacol. Sin. 30(6): 740-751 (2009), Bencherif et al., J. Pharmacol. Exp. Ther. 279: 1413 (1996), Lippiello et al., J. Pharmacol. Exp. Ther. 279: 1422 (1996), Damaj et al., J.

Pharmacol. Exp. Ther. 291 : 390 (1999); Chiari et al., Anesthesiology 91 : 1447 (1999), Lavand'homme and Eisenbach, Anesthesiology 91 : 1455 (1999), Holladay et al., J. Med. Chem. 40(28): 4169-94 (1997), Bannon et al., Science 279: 77 (1998), PCT WO 94/08992, PCT WO 96/31475, PCT WO 96/40682, and U.S. Patent Nos. 5,583,140 to Bencherif et al., 5,597,919 to Dull et al., 5,604,231 to Smith et al. and 5,852,041 to Cosford et al.

CNS Disorders

The compounds and their pharmaceutical compositions are useful in the treatment or prevention of a variety of CNS disorders, including neurodegenerative disorders, neuropsychiatric disorders, neurologic disorders, and addictions. The compounds and their pharmaceutical compositions can be used to treat or prevent cognitive deficits and dysfunctions, age-related and otherwise; attentional disorders and dementias, including those due to infectious agents or metabolic disturbances; to provide neuroprotection; to treat convulsions and multiple cerebral infarcts; to treat mood disorders, compulsions and addictive behaviors; to provide analgesia; to control inflammation, such as mediated by cytokines and nuclear factor kappa B; to treat inflammatory disorders; to provide pain relief; and to treat infections, as anti-infectious agents for treating bacterial, fungal, and viral infections. Among the disorders, diseases and conditions that the compounds and pharmaceutical compositions of the present invention can be used to treat or prevent are: age-associated memory impairment (AAMI), mild cognitive impairment (MCI), age-related cognitive decline (ARCD), pre-senile dementia, early onset Alzheimer's disease, senile dementia, dementia of the Alzheimer's type, Alzheimer's disease, cognitive impairment no dementia (CIND), Lewy body dementia, HIV-dementia, AIDS dementia complex, vascular dementia, Down syndrome, head trauma, traumatic brain injury (TBI), dementia pugilistica, Creutzfeld-Jacob Disease and prion diseases, stroke, central ischemia, peripheral ischemia, attention deficit disorder, attention deficit hyperactivity disorder, dyslexia, schizophrenia, schizophreniform disorder, schizoaffective disorder, cognitive dysfunction in schizophrenia, cognitive deficits in schizophrenia, Parkinsonism including Parkinson's disease, postencephalitic parkinsonism, parkinsonism-dementia of Gaum, frontotemporal dementia Parkinson's Type (FTDP), Pick's disease, Niemann-Pick's Disease, Huntington's Disease, Huntington's chorea, abnormal involuntary movements, dyskinesias, L-dopa induced dyskinesia, tardive dyskinesia, spastic dystonia, hyperkinesia, progressive supranuclear palsy, progressive supranuclear paresis, restless leg syndrome, Creutzfeld-Jakob disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), motor neuron diseases (MND), multiple system atrophy (MSA), corticobasal degeneration, Guillain-Barre Syndrome (GBS), and chronic inflammatory demyelinating polyneuropathy (CIDP), epilepsy, autosomal dominant nocturnal frontal lobe epilepsy, mania, anxiety, depression, premenstrual dysphoria, panic disorders, bulimia, anorexia, narcolepsy, excessive daytime sleepiness, bipolar disorders, generalized anxiety disorder, obsessive compulsive disorder, rage outbursts, conduct disorder, oppositional defiant disorder, Tourette's syndrome, autism, drug and alcohol addiction, tobacco addiction, compulsive overeating and sexual dysfunction.

In addition, the compounds and their pharmaceutical compositions are useful in the treatment of involuntary muscle contractions or relaxations involving movements of one or more body parts, whether such is a disorder itself or as a symptom of another disorder. In one embodiment, the compounds and their pharmaceutical compositions are useful in the treatment of one or more tremor, including but not limited to Essential tremor, Parkinsonian tremor, Cerebellar tremor (including intention tremor and titubation), Rubral tremor (Holmes' tremor), Dystonic tremor, Wilson's disease, Tremor in multiple sclerosis, Tremor resulting from a neurodegenerative disorder other than those specifically listed, Tremor resulting from basal ganglia disease other than Parkinson's disease and dystonia, Familial tremor other than essential tremor and Wilson's disease, Tremor resulting from peripheral neuropathy, Tremor resulting from stroke, Tremor resulting from traumatic brain injury, Tumor-induced tremor, Enhanced or exaggerated physiologic tremor, Tremor resulting from metabolic disorders, Tremor resulting from chronic kidney disease, Tremor resulting from liver failure, Tremor resulting from phenylketonuria, Orthostatic tremor, Psychogenic tremor, Drug- induced tremor, Tremor resulting from alcoholism (asterixis), and Tremor resulting from toxins (e.g. heavy metal poisoning).

Similarly, the compounds and their pharmaceutical compositions are useful in the treatment of sustained muscle contractions resulting in twisting or repetitive movements or abnormal postures, whether such is a disorder itself or as a symptom of another disorder. In another embodiment, the compounds and their pharmaceutical compositions are useful in the treatment of one or more dystonia, including but not limited to, Genetic or familial dystonia (one or more genes associated with various forms of dystonia), Myoclonic dystonia, General dystonia, lodiopathic torsion dystonia, Dystonia associated with Parkinson's disease, Dystonia associated with Wilson's disease, Focal dystonia such as,

blepharospasm, cervical dystonia or torticollis, cranial dystonia, oromandibular dystonia, or spasmodic dystonia, Multifocal dystonia, Cranial dystonia or Meige's syndrome, Segmental dystonia, Hemidystonia, Torsion dystonia, Paroxysmal dystonia, Dystonia resulting from trauma, Dystonia resulting from stroke, Tumor-induced dystonia, Dystonia resulting from infection, Dystonia resulting from oxygen deprivation, Dystonia resulting from toxins (e.g. heavy metal poisoning, carbon dioxide poisoning), and Drug-induced dystonia.

Cognitive impairments or dysfunctions may be associated with psychiatric disorders or conditions, such as schizophrenia and other psychotic disorders, including but not limited to psychotic disorder, schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, shared psychotic disorder, and psychotic disorders due to a general medical conditions, dementias and other cognitive disorders, including but not limited to mild cognitive impairment, pre-senile dementia, Alzheimer's disease, senile dementia, dementia of the Alzheimer's type, age-related memory impairment, Lewy body dementia, vascular dementia, AIDS dementia complex, dyslexia, Parkinsonism including Parkinson's disease, dyskinesias, levodopa-induced dyskinesias (LIDs), abnormal involuntary movements (AIMs), cognitive impairment and dementia of Parkinson's Disease, cognitive impairment of multiple sclerosis, cognitive impairment caused by traumatic brain injury, dementias due to other general medical conditions, anxiety disorders, including but not limited to panic disorder without agoraphobia, panic disorder with agoraphobia, agoraphobia without history of panic disorder, specific phobia, social phobia, obsessive- compulsive disorder, post-traumatic stress disorder, acute stress disorder, generalized anxiety disorder and generalized anxiety disorder due to a general medical condition, mood disorders, including but not limited to major depressive disorder, dysthymic disorder, bipolar depression, bipolar mania, bipolar I disorder, depression associated with manic, depressive or mixed episodes, bipolar II disorder, cyclothymic disorder, and mood disorders due to general medical conditions, sleep disorders, including but not limited to dyssomnia disorders, primary insomnia, primary hypersomnia, narcolepsy, parasomnia disorders, nightmare disorder, sleep terror disorder and sleepwalking disorder, mental retardation, learning disorders, motor skills disorders, communication disorders, pervasive developmental disorders, attention-deficit and disruptive behavior disorders, attention deficit disorder, attention deficit hyperactivity disorder, feeding and eating disorders of infancy, childhood, or adults, tic disorders, elimination disorders, substance- related disorders, including but not limited to substance dependence, substance abuse, substance intoxication, substance withdrawal, alcohol-related disorders, amphetamine or amphetamine-like-related disorders, caffeine-related disorders, cannabis-related disorders, cocaine-related disorders, hallucinogen-related disorders, inhalant-related disorders, nicotine-related disorders, opioid- related disorders, phencyclidine or phencyclidine-like-related disorders, and sedative-, hypnotic- or anxiolytic-related disorders, personality disorders, including but not limited to obsessive-compulsive personality disorder and impulse-control disorders. Cognitive performance may be assessed with a validated cognitive scale, such as, for example, the cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-cog). One measure of the effectiveness of the compounds of the present invention in improving cognition may include measuring a patient's degree of change according to such a scale.

Regarding compulsions and addictive behaviors, the compounds of the present invention may be used as a therapy for nicotine addiction and for other brain-reward disorders, such as substance abuse including alcohol addiction, illicit and prescription drug addiction, eating disorders, including obesity, and behavioral addictions, such as gambling, or other similar behavioral manifestations of addiction.

The above conditions and disorders are discussed in further detail, for example, in the American Psychiatric Association: Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision, Washington, DC, American Psychiatric Association, 2000. This Manual may also be referred to for greater detail on the symptoms and diagnostic features associated with substance use, abuse, and dependence. This Manual is updated and revised from time-to-time and the description of the diseases and conditions herein is intended to remain consistent with such revision.

Preferably, the treatment or prevention of diseases, disorders and conditions occurs without appreciable adverse side effects, including, for example, significant increases in blood pressure and heart rate, significant negative effects upon the gastro-intestinal tract, and significant effects upon skeletal muscle.

The compounds of the present invention, when employed in effective amounts, are believed to modulate the activity of the α4β2* and/or α6β2* NNRs without appreciable interaction with the nicotinic subtypes that characterize the human ganglia, as demonstrated by a lack of the ability to elicit nicotinic function in adrenal chromaffin tissue, or skeletal muscle, further demonstrated by a lack of the ability to elicit nicotinic function in cell preparations expressing muscle-type nicotinic receptors. Thus, these compounds are believed capable of treating or preventing diseases, disorders and conditions without eliciting significant side effects associated activity at ganglionic and neuromuscular sites. Thus, administration of the compounds is believed to provide a therapeutic window in which treatment of certain diseases, disorders and conditions is provided, and certain side effects are avoided. That is, an effective dose of the compound is believed sufficient to provide the desired effects upon the disease, disorder or condition, but is believed insufficient, namely is not at a high enough level, to provide undesirable side effects.

Thus, the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable salt thereof, for use in therapy, such as a therapy described above.

In yet another aspect the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in the treatment of a CNS disorder, such as a disorder, disease or condition described hereinabove.

Inflammation

The nervous system, primarily through the vagus nerve, is known to regulate the magnitude of the innate immune response by inhibiting the release of macrophage tumor necrosis factor (TNF). This physiological mechanism is known as the "cholinergic antiinflammatory pathway" (see, for example, Tracey, "The Inflammatory Reflex," Nature 420: 853-9 (2002)). Excessive inflammation and tumor necrosis factor synthesis cause morbidity and even mortality in a variety of diseases. These diseases include, but are not limited to, endotoxemia, rheumatoid arthritis, osteoarthritis, psoriasis, asthma, atherosclerosis, idiopathic pulmonary fibrosis, and inflammatory bowel disease.

Inflammatory conditions that can be treated or prevented by administering the compounds described herein include, but are not limited to, chronic and acute inflammation, psoriasis, endotoxemia, gout, acute pseudogout, acute gouty arthritis, arthritis, rheumatoid arthritis, osteoarthritis, allograft rejection, chronic transplant rejection, asthma,

atherosclerosis, mononuclear-phagocyte dependent lung injury, idiopathic pulmonary fibrosis, atopic dermatitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, acute chest syndrome in sickle cell disease, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, ulcers, ulcerative colitis, acute cholangitis, aphthous stomatitis, cachexia, pouchitis, glomerulonephritis, lupus nephritis, thrombosis, and graft vs. host reaction.

Inflammatory Response Associated with Bacterial and/or Viral Infection

Many bacterial and/or viral infections are associated with side effects brought on by the formation of toxins, and the body's natural response to the bacteria or virus and/or the toxins. As discussed above, the body's response to infection often involves generating a significant amount of TNF and/or other cytokines. The over-expression of these cytokines can result in significant injury, such as septic shock (when the bacteria is sepsis), endotoxic shock, urosepsis, viral pneumonitis and toxic shock syndrome.

Cytokine expression is mediated by NNRs, and can be inhibited by administering agonists or partial agonists of these receptors. Those compounds described herein that are agonists or partial agonists of these receptors can therefore be used to minimize the inflammatory response associated with bacterial infection, as well as viral and fungal infections. Examples of such bacterial infections include anthrax, botulism, and sepsis. Some of these compounds may also have antimicrobial properties.

These compounds can also be used as adjunct therapy in combination with existing therapies to manage bacterial, viral and fungal infections, such as antibiotics, antivirals and antifungals. Antitoxins can also be used to bind to toxins produced by the infectious agents and allow the bound toxins to pass through the body without generating an inflammatory response. Examples of antitoxins are disclosed, for example, in U.S. Patent No. 6,310,043 to Bundle et al. Other agents effective against bacterial and other toxins can be effective and their therapeutic effect can be complemented by co-administration with the compounds described herein.

Pain

The compounds can be administered to treat and/or prevent pain, including acute, neurologic, inflammatory, neuropathic and chronic pain. The compounds can be used in conjunction with opiates to minimize the likelihood of opiate addiction (e.g., morphine sparing therapy). The analgesic activity of compounds described herein can be demonstrated in models of persistent inflammatory pain and of neuropathic pain, performed as described in U.S. Published Patent Application No. 20010056084 A1 (Allgeier et al.) (e.g., mechanical hyperalgesia in the complete Freund's adjuvant rat model of inflammatory pain and mechanical hyperalgesia in the mouse partial sciatic nerve ligation model of neuropathic pain).

The analgesic effect is suitable for treating pain of various genesis or etiology, in particular in treating inflammatory pain and associated hyperalgesia, neuropathic pain and associated hyperalgesia, chronic pain (e.g., severe chronic pain, post-operative pain and pain associated with various conditions including cancer, angina, renal or biliary colic, menstruation, migraine, and gout). Inflammatory pain may be of diverse genesis, including arthritis and rheumatoid disease, teno-synovitis and vasculitis. Neuropathic pain includes trigeminal or herpetic neuralgia, neuropathies such as diabetic neuropathy pain, causalgia, low back pain and deafferentation syndromes such as brachial plexus avulsion.

Other Disorders

In addition to treating CNS disorders, inflammation, and neovascularization, and pain, the compounds of the present invention can be also used to prevent or treat certain other conditions, diseases, and disorders in which NNRs play a role. Examples include autoimmune disorders such as lupus, disorders associated with cytokine release, cachexia secondary to infection (e.g., as occurs in AIDS, AIDS related complex and neoplasia), obesity, pemphitis, urinary incontinence, overactive bladder, diarrhea, constipation, retinal diseases, infectious diseases, myasthenia, Eaton-Lambert syndrome, hypertension, preeclampsia, osteoporosis, vasoconstriction, vasodilatation, cardiac arrhythmias, type I diabetes, type II diabetes, bulimia, anorexia and sexual dysfunction, as well as those indications set forth in published PCT application WO 98/25619. The compounds of this invention can also be administered to treat convulsions such as those that are symptomatic of epilepsy, and to treat conditions such as syphillis and Creutzfeld-Jakob disease. Lastly, the compounds of this invention may be used to treat a variety of dermatological disorders, including but not limited to psoriasis, dermatitis, acne, pustulosis, vitilago, and the like. Diagnostic Uses

The compounds can be used in diagnostic compositions, such as probes, particularly when they are modified to include appropriate labels. The probes can be used, for example, to determine the relative number and/or function of specific receptors, particularly the α4β2* and/or a6-containing receptor subtypes. For this purpose the compounds of the present invention most preferably are labeled with a radioactive isotopic moiety such as ¹¹C, which can be detected using positron emission tomography (PET). A high specific activity is desired to visualize the selected receptor subtypes at non-saturating concentrations. The administered doses typically are below the toxic range and provide high contrast images. The compounds are expected to be capable of administration in non-toxic levels.

Determination of dose is carried out in a manner known to one skilled in the art of radiolabel imaging. See, for example, U.S. Patent No. 5,969, 144 to London et al.

The compounds can be administered using known techniques. See, for example,

U.S. Patent No. 5,969, 144 to London et al., as noted. The compounds can be administered in formulation compositions that incorporate other ingredients, such as those types of ingredients that are useful in formulating a diagnostic composition. Compounds useful in accordance with carrying out the present invention most preferably are employed in forms of high purity. See, U.S. Patent No. 5,853,696 to Elmalch et al.

After the compounds are administered to a subject (e.g., a human subject), the presence of that compound within the subject can be imaged and quantified by appropriate techniques in order to indicate the presence, quantity, and functionality of selected NNR subtypes. In addition to humans, the compounds can also be administered to animals, such as mice, rats, dogs, and monkeys. PET imaging can be carried out using any appropriate technique and apparatus. See Villemagne et al., In: Arneric et al. (Eds.) Neuronal Nicotinic Receptors: Pharmacology and Therapeutic Opportunities, 235-250 (1998) and U.S. Patent No. 5,853,696 to Elmalch et al., each herein incporated by reference, for a disclosure of representative imaging techniques.

The radiolabeled compounds bind with high affinity to selective NNR subtypes (e.g., α4β2* and/or a6-containing) and preferably exhibit negligible non-specific binding to other nicotinic cholinergic receptor subtypes (e.g., those receptor subtypes associated with muscle and ganglia). As such, the compounds can be used as agents for noninvasive imaging of nicotinic cholinergic receptor subtypes within the body of a subject, particularly within the brain for diagnosis associated with a variety of CNS diseases and disorders.

In one aspect, the diagnostic compositions can be used in a method to diagnose disease in a subject, such as a human patient. The method involves administering to that patient a detectably labeled compound as described herein, and detecting the binding of that compound to selected NNR subtypes (e.g., α4β2^* and/or a6-containing receptor subtypes). Those skilled in the art of using diagnostic tools, such as PET, can use the radiolabeled compounds described herein to diagnose a wide variety of conditions and disorders, including conditions and disorders associated with dysfunction of the central and autonomic nervous systems. Such disorders include a wide variety of CNS diseases and disorders, including Alzheimer's disease, Parkinson's disease, and schizophrenia. These and other representative diseases and disorders that can be evaluated include those that are set forth in U.S. Patent No. 5,952,339 to Bencherif et al.

In another aspect, the diagnostic compositions can be used in a method to monitor selective nicotinic receptor subtypes of a subject, such as a human patient. The method involves administering a detectably labeled compound as described herein to that patient and detecting the binding of that compound to selected nicotinic receptor subtypes namely, the α4β2* and/or a6-containing receptor subtypes.

Receptor Binding

The compounds of this invention can be used as reference ligands in binding assays for compounds which bind to NNR subtypes, particularly the α4β2* and/or a6-containing receptor subtypes. For this purpose the compounds of this invention are preferably labeled with a radioactive isotopic moiety such as ³H, or ¹⁴C.

EXAMPLES

The following examples are provided to illustrate the present invention, and should not be construed as limiting thereof. In these examples, all parts and percentages are by weight, unless otherwise noted.

Example 1 : Synthesis of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane (Compound A)

A three-neck flask, equipped with a mechanical stirrer, two reflux condensers, a cold finger, a temperature probe, a nitrogen inlet and an exhaust outlet (leading to a aqueous sodium bisulfite/sodium hydroxide trap), was charged with glutaryl chloride (350 g, 2.07 mol) followed by bromine (160 ml_, 3.12 mol). The resulting mixture was stirred under nitrogen and heated at gentle reflux, as the internal temperature gradually increased from 58°C to 91 °C over a period of 7 h. During the heating period and as consumption/loss of bromine was observed, additional bromine was added twice (first 90 ml_ and later 120 ml_). The reaction mixture was then allowed to gradually cool to ambient temperature while stirring under a nitrogen atmosphere overnight. Analysis of an aliquot of the reaction mixture quenched in methanol indicated complete conversion to dimethyl 2,4-dibromoglutarate (>98 % analyzed as the based on GCMS and LCMS). This 2,4-dibromoglutaryl chloride was used without further purification in the next step.

A three-neck flask, equipped with a mechanical stirrer, a nitrogen inlet, an addition funnel and a temperature probe, was charged with anhydrous ether (7 L) followed by benzyl alcohol (493 g, 4.56 mol). The solution was stirred under nitrogen and cooled in an ice water bath until an internal temperature of 8°C was reached. Then 2,4-dibromoglutaryl chloride was added through the addition funnel to the stirred solution (aided by an anhydrous ether rinse) over 1 to 1 .5 h during which time a maximum observed exotherm of 12°C occurred. Following the addition, the reaction mixture was stirred on the cold bath for 1 h (internal temperature observed to be 12°C). The reaction mixture was then allowed to stir under a nitrogen atmosphere while gradually warming to ambient temperature overnight. Analysis of an aliquot of the reaction mixture diluted in methanol indicated complete consumption of 2,4- dibromoglutaryl chloride (i.e., no dimethyl 2,4-dibromoglutarate was observed) based on LCMS. The reaction mixture was transferred to a separatory funnel (aided with an ether rinse) and the organic solution was washed twice with water, twice with 1 M aqueous sodium bisulfite, twice with saturated aqueous sodium bicarbonate (the second wash tested alkaline with pH paper) and finally once with saturated aqueous sodium chloride. The collected organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure affording intermediate dibenzyl 2,4-dibromoglutarate as a pale yellow oil (1 .034 kg, quantitative crude yield) which was used directly in the next step without further purification. ¹H NMR (CDCI₃, 300 MHz): δ 7.38 (s, 10H), 5.23 (s, 4H), 4.59- 4.41 (m, 2H), 2.97-2.64 (m, 2H).

A three-neck flask, equipped with a mechanical stirrer, a nitrogen inlet, a temperature probe, an addition funnel and a reflux condenser, was charged with a solution of crude dibenzyl 2,4-dibromoglutarate (Intermediate 3) (1 .034 kg, 2.199 mol) in dimethylformamide (2.93 L). The mixture was stirred under nitrogen at ambient temperature (internal temperature of 18°C) after which benzylamine (707 g, 720 ml_, 6.60 mol) was added by addition funnel in one portion (< 5 min addition time), during which time an exotherm to a maximum temperature of 55-60°C was observed. Immediately following the complete addition, the reaction mixture was heated at 93-95°C for 4.5 h. LCMS analysis of an aliquot of the reaction mixture diluted in methanol indicated complete consumption of intermediate 3 and formation of product (in addition to its trans counterpart). The heating was stopped and the reaction mixture was allowed to gradually cool to ambient temperature under nitrogen overnight. The reaction solution was then poured into a stirred biphasic mixture of 1 : 1 ethyl acetate/hexanes (8 L) and water (6 L). After agitation, the organic and aqueous phases were separated. The collected organic phase was washed successively with water, saturated aqueous sodium bicarbonate (wash tested alkaline with pH paper) and saturated aqueous sodium chloride. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure, providing 790 g of adark colored oil. The crude material was loaded onto a silica gel plug (loading aided with a wash of a minimal amount of dichloromethane) after which elution with 15% ethyl acetate/hexanes was performed. Concentration of selected fractions under reduced pressure to give 543 g of dibenzyl N-benzylazetidine-2,4-dicarboxylate with a 82: 17 ratio of cis (Intermediate 4):trans stereochemistry and an overall chemical purity of 92%. Crystallization from diethyl ether and air drying provided 310 g (36% overall yield from glutaryl chloride) of dibenzyl c/s-N- benzylazetidine-2,4-dicarboxylate (Intermediate 4) as a white solid (purity >96% based on LCMS). ¹H NMR (CDCI₃, 300 MHz): δ 7.30 (m, 10H), 5.07 (dd, 4H), 3.89 (s, 2H), 3.67 (dd, 2H), 2.55 (dd, 1 H), 2.36 (dd, 1 H); LCMS (m/z): 416 (M+1 ).

A three-neck flask, equipped with a mechanical stirrer, a temperature probe and a nitrogen inlet was charged with dibenzyl c/s-N-benzylazetidine-2,4-dicarboxylate (173 g, 416 mmol) followed by methanol (1 .14 L) and tetrahydrofuran (560 ml_). The resulting mixture was stirred at ambient temperature under nitrogen until homogeneity was obtained. The solution was then cooled in a dry ice/acetone bath until an internal temperature of -4°C was reached. Then sodium borohydride (79.0 g, 2.08 mol) was added in portions over 1-1 .5 h, during which time a slight exotherm to a maximum temperature of 6°C was observed.

Following the addition, the resulting mixture was stirred under nitrogen while gradually warming to ambient temperature overnight [Note: during warming, a secondary exotherm up to a maximum temperature of 27°C was observed and was controlled by the use of an ice bath]. LCMS analysis of an aliquot of the reaction mixture diluted in methanol indicated consumption of stating material and formation of product (plus benzyl alcohol). The reaction mixture was then cooled in an ice water bath (internal temperature of 8°C) and quenched by the drop-wise addition of water (100 mL) via addition funnel. The resulting mixture was concentrated under reduced pressure to remove the bulk of volatiles. The remainder was partitioned between 10% methanol/dichloromethane (500 mL) and water (200 mL). After phase separation, the organic (upper layer) and aqueous phases were collected. The aqueous phase was again extracted with 10% methanol/dichloromethane (2 x 500 mL; organic layer is now the lower phase). Analysis of an aliquot of the aqueous phase at this stage shows no product present. The combined organic extracts were washed successively with water and saturated aqueous sodium chloride. Analysis of this aqueous (water plus saturated aqueous sodium chloride) phase showed the presence of product (plus benzyl alcohol), so the aqueous phases were back-extracted with 10% methanol/dichloromethane (3 x). All the methanol/dichloromethane extracts were combined and dried over anhydrous sodium sulfate. After filtration and concentration under reduced pressure, 177 g of a yellow colored oil was obtained. Further concentration of this residue under high vacuum in a water bath at 75°C was performed to remove the bulk of benzyl alcohol. After constant weight was observed, 100 g of an oil remained. This material was dissolved in dichloromethane and concentrated under reduced pressure giving a semisolid. The semisolid was twice suspended in hexanes and subsequently concentrated under reduced pressure. The remaining solid was suspended in hexanes (260 mL) and the resulting suspension was stirred overnight. The suspension was then filtered and the collected solids were washed with hexanes (2 x). LCMS analysis of a sample of the solid dissolved in methanol indicated a product purity of -90% (contaminated with benzyl alcohol). The solids were again suspended in hexanes (200 mL) and the suspension stirred (on rotovap) for 1 hour at 30°C. The suspension was filtered, and the collected solids were washed with additional hexanes (1 x). LCMS analysis of a sample of the solid dissolved in methanol indicated a product purity >96%. The solids were further air dried to constant weight, affording 81 .9 g (95% yield) of c/s-N-benzyl-2,4-bis(hydroxymethyl)azetidine (intermediate 5) as a white solid. ¹H NMR (CDCI₃, 300 MHz): δ 7.28 (m, 5H), 4.24 (dd, 2H), 3.63 (s, 2H), 3.12 (dd, 4H), 3.05 (m, 2H), 2.00 (dd, 1 H), 1 .57 (dd, 1 H); LCMS (m/z): 208 (M+1 ).

A three-neck flask, equipped with a mechanical stirrer, a temperature probe, a nitrogen inlet and an addition funnel, was charged with c/s-N-benzyl-2,4- bis(hydroxymethyl)azetidine (81 .9 g, 395 mmol) followed by anhydrous dichloromethane (820 mL). The resulting solution was stirred under nitrogen, after which triethylamine (160 g, 220 mL, 1 .58 mol) was added in one portion. The resulting solution was cooled in a dry ice/acetone bath until an internal temperature of -6°C was reached. Then methanesulfonyl chloride (109 g, 73.4 mL, 948 mmol) was added drop-wise by addition funnel over 45 min, during which time an exotherm occurred up to a maximum observed temperature of 4°C. Following the addition, the reaction mixture was allowed to gradually warm to ambient temperature while stirring under nitrogen overnight. LCMS analysis of an aliquot of the reaction mixture diluted in acetonitrile indicated consumption of starting material and formation of product. To the reaction mixture was added water (250 mL) drop-wise followed by saturated aqueous sodium bicarbonate (250 mL). The resulting biphasic mixture was stirred vigorously for 20 min and transferred to a separatory funnel, aided by a

dichloromethane rinse. The organic and aqueous phases were separated and the organic layer was then washed with saturated aqueous sodium chloride. After phase separation (slow), the organic layer was collected, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, leaving 143 g (100% yield) of the bis-mesylate (Intermediate 6), as a brown colored oil, which was used without purification in the next step. ¹H NMR (CDCIa, 300 MHz): δ 7.33 (m, 5H), 3.97 (ddd, 4H), 3.74 (s, 2H), 3.44 (m, 2H), 2.92 (s, 6H), 2.28 (m, 1 H), 1 .91 (m, 1 H); LCMS (m/z): 364 (M+1 ).

A three-neck flask, equipped with a mechanical stirrer, a temperature probe, a nitrogen inlet and a reflux condenser, was charged with cyclopropanecarboxamide (36.8 g, 432 mmol) under an atmosphere of nitrogen. Anhydrous 1 -methyl-2-pyrrolidinone (N- methyl-2-pyrrolidone) (900 mL) was added and the resulting mixture was mechanically stirred under nitrogen while cooling in an ice water bath (internal temperature of 5°C). Then sodium hydride (39.4 g of a 60% dispersion in mineral oil, 980 mmol) was added in portions over 30 min, as the temperature varied between 5 and 12°C. The resulting mixture was stirred on the cold bath for an additional 10 min. The cold bath was allowed to warm to ambient temperature (over 30 min), at which point a slight exotherm (to a maximum temperature of 25°C) and gas evolution were observed. The resulting mixture was stirred at ambient temperature for an additional 50 min (no further gas evolution), after which a solution of the bis-mesylate (Intermediate 6) (143 g, 393 mmol) in anhydrous 1 -methyl-2- pyrrolidinone (600 mL) was added to the suspension in one lot. The flask containing crude Intermediate 6 was rinsed with additional anhydrous 1 -methyl-2-pyrrolidinone (2 x 100 mL), each rinse being added to the reaction suspension. The resulting mixture was then heated at 68°C for 3 h. LCMS analysis of an aliquot, withdrawn after 2 h of heating and quenched in moist acetonitrile, indicated that >96% of Intermediate 6 had been consumed and that desired product predominated. The reaction mixture was allowed to cool under nitrogen to ambient temperature overnight. The viscous solution was quenched, in an ice water bath cooling, by the addition of water (-10 mL) and diluted with methyl t-butyl ether (100 mL) and stirred for 10 min. The mixture was then transferred to a separatory funnel containing 5% aqueous sodium chloride (4.0 L) and 25% ethyl acetate/methyl t-butyl ether (3.0 L). The resulting mixture was agitated. After phase separation, the organic and aqueous layers were drawn off. The aqueous phase was extracted with additional 25% ethyl acetate/methyl t-butyl ether (2 x 3.0 L). The combined organic extracts were washed with water (1 x 3.0 L) and then saturated aqueous sodium chloride (1 x 3.0 L). The organic phase was then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, affording 86 g of brown colored oil. This residue was purified by silica gel chromatography, eluting initially with 0 to 40% ethyl acetate/hexanes and finally with 60 to 100% ethyl

acetate/hexanes + 0.5% triethylamine. Concentration of selected fractions gave 56 g of 6- benzyl-3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1 .1]heptane with HPLC purity of 78%. This was used in the next step without further purification. A three-neck flask, equipped with a mechanical stirrer, a temperature probe, a reflux condenser and a nitrogen inlet, was charged with a solution of 6-benzyl-3- cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane (56 g of 78%, 220 mmol) in ethanol (200 proof, 670 ml_) under nitrogen. Then 10% Pd/C (33.6 g, wet) was added followed by ammonium formate (82.6 g, 1.31 mol). The resulting mixture was rigorously stirred under a nitrogen atmosphere and gradually heated at 60 to 66°C for 6 h, an additional 20 g of 10% Pd/C and 60 g of ammonium formate being added after -3.5 h. The reaction was then heated (between 66°C and 71 °C for 6 h) and periodically monitored by LCMS, as more reagents were added in several portions (total additional ammonium formate = 96 g; total additional 10% Pd/C = 21 g). An analysis of an aliquot of the reaction mixture then indicated consumption of starting material. Heating was stopped and the reaction mixture was allowed to gradually cool to ambient temperature under nitrogen overnight. The reaction mixture was then filtered through a bed of diatomaceous earth. The filter cake was subsequently washed with methanol (4 x), and the combined filtrates were concentrated under reduced pressure, providing 35 g of a pale yellow colored oil. The material was purified by silica gel chromatography, eluting with 0 to 60% DCMA80 in dichloromethane. [Note: DCMA80 is a 80:18:2 mixture of dichloromethane, methanol, and aqueous ammonium hydroxide]. Selected fractions were combined and concentrated under reduced pressure, affording 9.2 g of a nearly white solid with HPLC purity of 97.7 %. The purity of this material could be increased to 98.7% (HPLC) by trituration with methyl t-butyl ether. Less pure fractions from the chromatography were also concentrated, yielding an additional 1 1 g of material (~ 85% purity by HPLC). ¹H NMR (D₂0, 300 MHz): δ 4.02 (s, 2H), 3.73 (m, 2H), 3.64 (dd, 2H), 2.66 (m, 1 H), 1.92 (m, 1 H), 1.52 (d, 1 H), 0.88 (m, 4H); LCMS (m/z): 167 (M+1 ).

Example 2: Synthesis of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane hemigalactarate monohydrate {3,6-diazabicyclo[3.1.1]heptan-3- yl(cyclopropyl)methanone hemigalactarate monohydrate}

A three neck round bottom flask, equipped with a mechanical stirrer, a temperature probe, a reflux condenser and a nitrogen inlet, was charged with 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1]heptane (132 g, 0.794 mol, representing multiple runs of the chemistry shown in Example 2) under nitrogen. Ethanol (200 proof, 925 mL) and water (1 10 mL) were added to the reaction flask, and the mixture was stirred at ambient temperature until solution was obtained. Then, galactaric acid (86.0 g, 0.397 mol) was added and the resulting mixture was heated at 50°C under nitrogen for 2 h. Heating was then stopped, and the resulting thick slurry was allowed to cool to ambient temperature while stirring under nitrogen overnight. The mixture was then cooled in an ice water bath to 0°C and suction filtered. The collected solids were air dried briefly and then dissolved in hot water (1000 mL; 60°C). The solution was filtered while hot to remove a small amount of insoluble material and then concentrated to near dryness. To this residue was added ethanol (200 proof, 800 ml_), and the mixture was stirred at 50°C for 30 min and then cooled to 0°C in an ice water bath where it remained for 45 min. The slurry was filtered, and the collected solids were dried under vacuum overnight. Further drying of the solid was performed under vacuum at 56°C to constant weight (over a period of 4 h), giving 208 g (95% yield) of 3-cyclopropylcarbonyl-3,6- diazabicyclo[3.1.1]heptane hemigalactarate monohydrate as a white solid (HPLC purity 99%). ¹H NMR (D₂0, 400 MHz): δ 4.39 (m, 2H), 4.20 (dd, 2H), 4.09 (s, 1 H), 3.88 (d, 1 H), 3.79 (s, 1 H), 3.74 (d, 1 H), 2.93 (m, 1 H), 1.77 (m, 2H), 0.80 (m, 4H); LCMS (m/z): 167 (M+1 ); Karl Fischer analysis of the dried material indicated 6.5% water content (corresponds to monohydrate stoichiometry).

Example 3: Synthesis of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane hydrochloride, using amide coupling procedure

To tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (50 mg, 0.25 mmol) in a 25 ml_ round bottom flask were added cyclopropanecarboxylic acid (26 mg, 24 μΙ_, 0.30 mmol), triethylamine (70 μΙ_, 0.50 mmol), dichloromethane (5 ml_) and O-benzotriazol-1-yl- tetramethyluronium hexafluorophosphate (191 mg, 0.50 mmol). The reaction was stirred at ambient temperature for 2 h. Saturated ammonium chloride (5 ml_) was added, and the reaction mixture was stirred for 30 min. The mixture was then passed through a phase extractor, and the solvent was removed under vacuum. The crude mixture was dissolved in 3 ml_ of ethyl acetate and concentrated HCI (1 ml_) was added. This mixture was stirred for 2 h. The solvent was removed under vacuum, and the residue was passed through a silica- based cation exchange column, which was pre-washed with 2 ml_ of methanol followed by 2 ml_ of dichloromethane/methanol (1 :1 ). The residue was taken up in 1 ml_

dichloromethane/methanol and passed through the column, eluting with 3 ml_

dichloromethane/methanol (1 :1 ) followed by methanol/ammonia (7M). The eluates were concentrated, and the residue was purified by silica gel column chromatography, eluting with a gradient of CMA90 (chloroform/methanol/aqueous ammonium hydroxide 90:9:1 ) in chloroform. Selected fractions were concentrated under vacuum, and the residue was combined with 1 M hydrogen chloride in methanol (2 ml_). The resulting mixture was filtered through a phase extractor (to remove any fine particles), and the solvent was removed under vacuum, leaving 3,6-diazabicyclo[3.1.1]heptan-3-yl(cyclopropyl)methanone hydrochloride, as an oil (14 mg, 27%). ¹H NMR (CD₃OD, 400 MHz): δ 4.49 (m, 2H), 4.32 (dd, 2H), 4.01 (d, 1 H), 3.88 (d, 1 H), 3.06 (m, 1 H), 1.94 (m, 2H), 0.93 (m, 4H). Note: While this material failed to crystallize, other samples of the hydrochloride salt were induced to crystallize (after trituration with acetone). The resulting solid is hygroscopic. Example 4: Preparation of Additional Salts

Pharmaceutically acceptable acids were selected for inclusion in a salt selection study as outlined in PCT7 US2012/028691 , incorporated by reference hereinabove. In summary, to a solution of 30.0 mg (0.165 mmol) of free base in the selected solvent at 50°C, the corresponding acid (0.5 equivalent or 1.1 equivalent depending on the number and strength of the acidic moieties of the acid) was added as a solution or suspension.

The mixture was stirred at 50°C for 1 h, then slowly cooled to 0°C overnight

(0.1 °C/min). The solids were filtered and analyzed. If no solid was produced, the corresponding solution was allowed to evaporate slowly at ambient conditions.

In the case of clear solutions, gums, or oils, these were cooled to -20°C, and allowed to evaporate slowly to encourage crystallization. Thereafter, anti-solvent (co-solvent; methyl t-butyl ether) was then added, if oils/gums would not crystallize; followed by trituration with acetone. With this added effort, many salts gave a precipitate when placed in isopropyl acetate. Initially, a salt screen was performed using isopropyl acetate, however, for those experiments which produced non isolatable salts, a further salt screen was performed using isopropyl alcohol and 2-butanone (methyl ethyl ketone) (see the two tables below). All solids were filtered and analyzed by XRPD.

The hemigalactarate monohydrochloide salt appears to have preferential characteristics for commercialization as a drug product.

Example 5: Harmaline-induced Tremor Assay

The purpose of the study was to test Compound A in the harmaline-induced tremor assay in ICR mice. This study was performed at PsychoGenics, Inc., headquartered at 765 Old Saw Mill Road, Tarrytown, New York.

Test Procedures: Animals

Male ICR mice from Taconic (Germantown, NY) were used in this study. Upon receipt, mice were group housed in OPTI mouse ventilated cages. All animals remained group housed during the duration of the study. All mice were acclimated to the colony room for at least one week prior to testing and subsequently tested at an average age of 10 weeks. During the period of acclimation, mice were examined on a regular basis, handled, and weighed to assure adequate health and suitability. Mice were maintained on a 12/12 light/dark cycle. The room temperature was maintained between 20 and 23°C with a relative humidity maintained between 30% and 70%. Chow and water were provided ad libitum for the duration of the study. In each test, animals were randomly assigned across treatment groups.

Test Compounds

The following compounds were used: Harmaline {Sigma, Lot 026K1538; 30 mg/kg) was dissolved in sterile saline and administered s.c.

Compound A (0.01. 0.1 and 1 mg/kg) was administered ip twice daily for 7 days prior to testing at a dose volume of 10 ml/kg. On day 8, Compound A was administered 20 minutes prior to harmaline administration.

Reference Compound: Propranolol HCI {Sigma; 10 mg/kg) was dissolved in sterile saline and administered i.p. 20 minutes prior to harmaline administration. Mice in this treatment group will receive 7 days of vehicle injections (ip; BID).

Treatment Groups

Ten mice were used in each treatment group for a total of 50 mice.

1. Sub-Chronic Vehicle + Harmaline (30 mg/kg)

2. Acute Propranolol (10 mg/kg) + Harmaline (30 mg/kg)

3. Sub-Chronic TI-314049 (0.01 mg/kg) + Harmaline (30 mg/kg)

4. Sub-Chronic TI-314049 (0.1 mg/kg) + Harmaline (30 mg/kg)

5. Sub-Chronic TI-314049 (1 mg/kg) + Harmaline (30 mg/kg)

Test procedure: Harmaline-induced tremors

Group-housed mice were brought to the experimental room for at least 1 h acclimation prior to testing. During test trials, the recorded frequencies (1-64 hertz) of activity and the number of tremor events were captured electronically.

Data were analyzed by the tremor monitor software (San Diego Instruments) in a two part process. Using a Fast Fourier Transform (FFT), an output is provided showing the percentage of activity (energy) recorded at each frequency. A center frequency of activity between 14 - 15 Hz is chosen, along with a bandwidth of 10 Hz. Using these parameters, tremor events were tabulated as short, long, and total events. A long event is defined as being greater than 0.5 seconds in duration, and a short event as greater than 0.3 but less than 0.5 seconds in duration.

Mice were injected with either vehicle, propranolol, or Compound A and placed in separate holding cages for 20 minutes following which mice were injected with harmaline (30 mg/kg) and placed inside the Tremor Monitor (San Diego Instruments, SDI) chamber for a 10 minute acclimation period. After habituation, tremor activity of the mice was measured for approximately 8 min. Chambers were cleaned with alcohol following each run.

Statistical Analysis

Data were analyzed by analysis of variance (ANOVA) followed by Fisher PLSD post- hoc analysis. An effect is considered significant if p < 0.05. Statistical outliers that fell above or below 2 standard deviations from the mean were removed from the final analysis.

Results

Compound A significantly reduces harmaline-induced tremor events in this mouse model of essential tremor. Figure 1 graphically depicts the effects of acute propranolol and sub-chronic Compound A administration on harmaline-induced tremor events. The asterisks (^*p<0.05) indicate a significant difference in short (0.3-0.5 seconds in duration), long (>0.5 seconds) and total tremor events compared to saline vehicle for respective measures. The specific pharmacological responses observed may vary according to and depending on the particular active compound selected or whether there are present pharmaceutical carriers, as well as the type of formulation and mode of administration employed, and such expected variations or differences in the results are contemplated in accordance with practice of the present invention.

Although specific embodiments of the present invention are herein illustrated and described in detail, the invention is not limited thereto. The above detailed descriptions are provided as exemplary of the present invention and should not be construed as constituting any limitation of the invention. Modifications will be obvious to those skilled in the art, and all modifications that do not depart from the spirit of the invention are intended to be included with the scope of the appended claims.

Claims

What is claimed is:

1. A method for treating one or more tremor comprising administering 3- cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane or a pharmaceutically acceptable salt thereof.

2. Use of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1 Jheptane or a pharmaceutically acceptable salt thereof in manufacture of a medicament for the treatment of one or more tremor.

3. A compound 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1 Jheptane or a

pharmaceutically acceptable salt thereof for use in treating one or more tremor.

4. A compound 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1 Jheptane or a

pharmaceutically acceptable salt thereof for use in the preparation of a medicament to treat one or more tremor.

5. The method, use, or compound of claims 1 - 4, wherein the tremor is one or more of Essential tremor, Parkinsonian tremor, Cerebellar tremor (including intention tremor and titubation), Rubral tremor (Holmes' tremor), Dystonic tremor, Wilson's disease,

Tremor in multiple sclerosis, Tremor resulting from a neurodegenerative disorder other than those specifically listed, Tremor resulting from basal ganglia disease other than Parkinson's disease and dystonia, Familial tremor other than essential tremor and Wilson's disease, Tremor resulting from peripheral neuropathy, Tremor resulting from stroke, Tremor resulting from traumatic brain injury, Tumor-induced tremor,

Enhanced or exaggerated physiologic tremor, Tremor resulting from metabolic disorders, Tremor resulting from chronic kidney disease, Tremor resulting from liver failure, Tremor resulting from phenylketonuria, Orthostatic tremor, Psychogenic tremor, Drug-induced tremor, Tremor resulting from alcoholism (asterixis), and Tremor resulting from toxins (e.g. heavy metal poisoning).

6. A method for treating one or more dystonia comprising administering 3- cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1]heptane or a pharmaceutically acceptable salt thereof.

7. Use of 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1 Jheptane or a pharmaceutically acceptable salt thereof in manufacture of a medicament for the treatment of one or more dystonia.

8. A compound 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1 Jheptane or a

pharmaceutically acceptable salt thereof for use in treating one or more dystonia.

9. A compound 3-cyclopropylcarbonyl-3,6-diazabicyclo[3.1.1 Jheptane or a

pharmaceutically acceptable salt thereof for use in the preparation of a medicament to treat one or more dystonia.

10. The method, use, or compound of claims 6 - 9, wherein the dystonia is one or more of Genetic or familial dystonia, Myoclonic dystonia, General dystonia, lodiopathic torsion dystonia, Dystonia associated with Parkinson's disease, Dystonia associated with Wilson's disease, Focal dystonia such as, blepharospasm, cervical dystonia or torticollis, cranial dystonia, oromandibular dystonia, or spasmodic dystonia, Multifocal dystonia, Cranial dystonia or Meige's syndrome, Segmental dystonia, Hemidystonia, Torsion dystonia, Paroxysmal dystonia, Dystonia resulting from trauma, Dystonia resulting from stroke, Tumor-induced dystonia, Dystonia resulting from infection, Dystonia resulting from oxygen deprivation, Dystonia resulting from toxins (e.g.

heavy metal poisoning, carbon dioxide poisoning), and Drug-induced dystonia.