AU2015258306A1 - Opsin-binding ligands, compositions and methods of use - Google Patents

Abstract

OPSIN BINDING LIGANDS, COMPOSITIONS AND METHODS OF USE Abstract Figure 1 Compounds and compositions of said compounds along with methods of use of compounds are disclosed for treating ophthalmic conditions related to mislocalization of opsin proteins, the misfolding of mutant opsin proteins and the production of toxic visual cycle products that accumulate in the eye. Compounds and compositions useful in the these methods, either alone or in combination with other therapeutic agents, are also described.

Description

1 OPSIN-BINDING LIGANDS, COMPOSITIONS AND METHODS OF USE 5 PRIORITY CLAIM This application claims priority of U.S. Provisional Application 61/268,757, filed 16 June 2009, the disclosure of which is hereby incorporated by reference in its entirety. 10 FIELD OF THE INVENTION The present invention relates to compounds and compositions thereof 15 for use in the treatment and/or prevention of ophthalmic diseases. BACKGROUND OF THE INVENTION 20 A diminished visual acuity or total loss of vision may result from a number of eye diseases or disorders caused by dysfunction of tissues or structures in the anterior segment of the eye and/or posterior segment of the eye. Of those that occur as a consequence of a dysfunction in the anterior segment, aberrations in the visual cycle are often involved. The visual cycle 25 (also frequently referred to as the retinoid cycle) comprises a series of light driven and/or enzyme catalyzed reactions whereby a light-sensitive chromophore (called rhodopsin) is formed by covalent bonding between the protein opsin and the retinoid agent 11-cis-retinal and subsequently, upon exposure to light, the 1 1-cis-retinal is converted to all-trans-retinal, which can 30 then be regenerated into 11-cis-retinal to again interact with opsin. A number of visual, ophthalmic, problems can arise due to interference with this cycle. It 2 is now understood that at least some of these problems are due to improper protein folding, such as that of the protein opsin. The main light and dark photoreceptor in the mammalian eye is the rod 5 cell, which contains a folded membrane containing protein molecules that can be sensitive to light, the main one being opsin. Like other proteins present in mammalian cells, opsin is synthesized in the endoplasmic reticulum (i.e., on ribosomes) of the cytoplasm and then conducted to the cell membrane of rod cells. In some cases, such as due to genetic defects and mutation of the opsin 10 protein, opsin can exhibit improper folding to form a conformation that either fails to properly insert into the membrane of the rod cell or else inserts but then fails to properly react with 11-cis-retinal to form native rhodopsin. In either case, the result is moderate to severe interference with visual perception in the animal so afflicted. 15 Among the diseases and conditions linked to improper opsin folding is retinitis pigmentosa (RP), a progressive ocular-neurodegenerative disease (or group of diseases) that affects an estimated 1 to 2 million people worldwide. In RP, photoreceptor cells in the retina are damaged or destroyed, leading to 20 loss of peripheral vision (i.e., tunnel vision) and subsequent partial or near total blindness. In the American population the most common defect occurs as a result of replacement of a proline residue by a histidine residue at amino acid 25 number 23 in the opsin polypeptide chain (dubbed "P23H"), caused by a mutation in the gene for opsin. The result is production of a destabilized form of the protein, which is misfolded and aggregates in the cytoplasm rather than being transported to the cell surface. Like many other protein conformational diseases (PCDs), the clinically common P23H opsin mutant associated with 30 autosomal dominant RP is misfolded and retained intracellularly. The aggregation of the misfolded protein is believed to result in photoreceptor damage and cell death.

3 Recent studies have identified small molecules that stabilize misfolded mutant proteins associated with disease. Some of these, dubbed "chemical chaperones," stabilize proteins non-specifically. Examples of these include glycerol and trimethylamine oxide. These are not very desirable for treating 5 ophthalmic disease because such treatment usually requires high .dosages that may cause toxic side effects. Other agents, dubbed "pharmacological chaperones," (which include native ligands and substrate analogs) act to stabilize the protein by binding to specific sites and have been identified for many misfolded proteins, e.g., G-protein coupled receptors. Opsin is an 10 example of a G-protein coupled receptor and its canonical pharmacological chaperones include the class of compounds referred to as retinoids. Thus, certain retinoid compounds have been shown to stabilize mutant opsin proteins (see, for example, U.S. Patent Pub. 2004-0242704, as well as Noorwez et al., J. Biol. Chem., 279(16): 16278-16284 (2004)). 15 The visual cycle comprises a series of enzyme catalyzed reactions, usually initiated by a light impulse, whereby the visual chromophore of rhodopsin, consisting of opsin protein bound covalently to 11-cis-retinal, is converted to an all-trans-isomer that is subsequently released from the 20 activated rhodopsin to form opsin and the all-trans-retinal product. This part of the visual cycle occurs in the outer portion of the rod cells of the retina of the eye. Subsequent parts of the cycle occur in the retinal pigmented epithelium (RPE). Components of this cycle include various enzymes, such as dehydrogenases and isomerases, as well as transport proteins for conveying 25 materials between the RPE and the rod cells. As a result of the visual cycle, various products are produced, called visual cycle products. One of these is all-trans-retinal produced in the rod cells as a direct result of light impulses contacting the 11-cis-retinal moiety of 30 rhodopsin. All-trans-retinal, after release from the activated rhodopsin, can be regenerated back into 11-cis-retinal or can react with an additional molecule of all-trans-retinal and a molecule of phosphatidylethanolamine to produce N retinylidene-N-retinylethanolamine (dubbed "A2E"), an orange-emitting 4 fluorophore that can subsequently collect in the rod cells and in the retina pigmented epithelium (RPE). As A2E builds up (as a normal consequence of the visual cycle) it can also be converted into lipofuscin, a toxic substance that has been implicated in several abnormalities, including ophthalmic conditions 5 such as wet and dry age related macular degeneration (ARMD). A2E can also prove toxic to the RPE and has been associated with dry ARMD. Because the build-up of toxic visual cycle products is a normal part of the physiological process, it is likely that all mammals, especially all humans, 10 possess such an accumulation to some extent throughout life. However, during surgical procedures on the eye, especially on the retina, where strong light is required over an extended period, for example, near the end of cataract surgery and while implanting the new lens, these otherwise natural processes can cause toxicity because of the build-up of natural products of 15 the visual cycle. Additionally, excessive rhodopsin activation as a result of bright light stimulation can cause photoreceptor cell apoptosis via an AP-1 transcription factor dependent mechanism. Because of this, there is a need for agents that can be administered prior to, during or after (or any combination of these) the surgical process and that has the effect of inhibiting 20 rhodopsin activation as well as reducing the production of visual cycle products that would otherwise accumulate and result in toxicity to the eye, especially to the retina. The present invention answers this need by providing small molecules 25 which noncovalently bind to opsin or mutated forms of opsin for treating and/or amelioration such conditions, if not preventing them completely. Importantly, such agents are not natural retinoids and thus are not tightly controlled for entrance into the rod cells, where mutated forms of opsin are synthesized and/or visual cycle products otherwise accumulate. Therefore, 30 such agents can essentially be titrated in as needed for facilitating the proper folding trafficking of mutated opsins to the cell membrane or prevention of rhodopsin activation that can lead to the excessive build-up of visual cycle products like all-trans-retinal that in turn can lead to toxic metabolic products.

5 Such compounds may compete with 11-cis-retinal to reduce all-trans-retinal by tying up the retinal binding pocket of opsin to prevent excessive all-trans retinal build up. Thus, the compounds provided by the present invention have the advantage that they do not directly inhibit the enzymatic processes by 5 which 11-cis-retinal is produced in the eye (thus not contributing to retinal degeneration). Instead, the formation of all-trans-retinal is limited and thereby the formation of A2E is reduced. Finally, by limiting the ability of 11-cis-retinal to combine with opsin to form rhodopsin, rhodopsin activation caused by bright light stimulation especially during ophthalmic surgery is also diminished 10 thus preventing the photocell death that results. Mislocalization of photoreceptor cell visual pigment proteins (opsins) can occur in various ocular diseases, and also with normal aging. In both cases the accumulation of mislocalized opsin leads to the decline in viability of 15 photoreceptor cells. With time this mislocalized opsin accumulation leads to rod and cone cell death, retinal degeneration, and loss of vision. The present invention solves this problem by providing a method of correcting mislocalized opsin within a photoreceptor cell by contacting a mislocalized opsin protein with an opsin-binding agent that binds reversibly and/or non-covalently to said 20 mislocalized opsin protein, and promotes the appropriate intracellular processing and transport of said opsin protein. This correction of mislocalization relieves photoreceptor cell stress, preventing decline in viability and death of photoreceptor cells in various diseases of vision loss, and in normal age-related decline in dim-light and peripheral rod-mediated 25 vision, central cone-mediated vision, and loss of night vision. Computer-assisted molecular docking has lead to the successful discovery of novel ligands for more than 30 targets (Shoichet et al., Curr. Opin. in Chem. Biol. 6: 439-46 (2002)). This strategy has been applied 30 primarily to enzymes, such as aldose reductase (Iwata et al., J. Med. Chem. 44: 1718-28 (2001)), Bcl-2, matriptase (Enyedy et al., J. Med. Chem. 44: 1349-55 (2001)), adenovirus protease (Pang et al., FEBS Letters 502: 93-97 (2001)), AmpC fl-lactamase, carbonic anhydrase (Gruneberg et al., J. Med.

6 Chem. 45: 3588-602 (2002)), HPRTase (Freymann et al., Chemistry & Biology 7: 957-68 (2000)), dihydrodipicolinate (Paiva et al., Biochimica Biophysica Acta 1545: 67-77 (2001)) and Cdk4 (Honma et al., J. Med. Chem. 44: 4615-27 (2001)). Improvements in docking algorithms and multiprocessor 5 resources have improved the technique of computer-assisted molecular docking such that it can now be applied to more challenging problems. For example, this approach has recently been applied to defining small molecules that target protein-protein interfaces, which are relatively broad and flat compared to easily targeted enzyme active sites. 10 More recently, a new computational technique defining the thermodynamic properties and phase behavior of water in confined regions of protein pockets has been developed (Young et. al., PNAS 104: 808-13 (2007)). The algorithm developed has been utilized to characterize the 15 solvation of protein pockets. The molecular dynamics simulations and solvent analysis techniques have characterized the solvation of hydrophobic enclosures and correlated hydrogen bonds as inducing atypical entropic and enthalpic penalties of hydration which stabilize the protein-ligand complex with respect to the independently solvated ligand and protein. These criteria, 20 commonly referred to as the water map, have been used to rationalize Factor Xa ligand binding (Abel et. al., JACS 130: 2817-31 (2008)). BRIEF SUMMARY OF THE INVENTION 25 In one aspect, the present invention provides compounds having the structure of Formula 1, including pharmaceutically acceptable salts, solvates and hydrates thereof, and compositions of said compounds: 30 A-B-Q-V Formula I wherein A, B, Q, and V are as described elsewhere herein.

7 In another aspect, the present invention provides compounds having the structure of Formula 11, A-B-Q-N2 P W Formula Il 5 wherein A, B, Q, W and p are as described elsewhere herein, including pharmaceutically acceptable salts, solvates and hydrates thereof, and compositions of said compounds. 10 In a related aspect, the present invention relates to a method of inhibiting the formation or accumulation of a visual cycle product, comprising contacting an opsin protein with a compound recited herein to inhibit formation of said visual cycle product relative to when said contacting does not occur. 15 In a further aspect, the present invention relates to a method to reduce the light toxicity associated with ophthalmic surgery by preventing rhodopsin regeneration during surgery to a mammalian eye and/or prevent or slow the formation of toxic visual cycle products by fractionally preventing rhodopsin formation during periods of light activation thereby providing a treatment of 20 ocular conditions associated with the build up of visual products such as wet or dry ARMD. In yet a further aspect, the present invention relates to a method of correcting the proper folding and trafficking of mutated opsin proteins, 25 comprising contacting a mutated opsin protein with a compound that stabilizes the proper three dimensional conformation of the protein relative to when said contacting does not occur wherein the compound has the structure of Formula I and/or Formula II including pharmaceutically acceptable salts, solvates and hydrates thereof. 30 8 In one embodiment, the ligand selectively binds reversibly or non covalently to opsin. In another embodiment, the ligand binds at or near the 11-cis- retinal binding pocket of the opsin protein. In yet another embodiment, the ligand binds to the opsin protein so as to inhibit or slow the covalent 5 binding of 11-cis-retinal to the opsin protein when the 11-cis-retinal is contacted with the opsin protein in the presence of the ligand. In yet another embodiment, the ligand binds to the opsin in the retinal binding pocket of opsin protein or disrupts 11-cis-retinal binding to the retinal binding pocket of opsin. In yet another embodiment, the ligand binds to the opsin protein so as 10 to inhibit covalent binding of 11-cis-retinal to the opsin protein. In yet another embodiment, the mammal is a human being. In yet another embodiment, slowing or halting the progression of wet or dry ARMD is associated with reducing the level of a visual cycle product, for 15 example, a visual cycle product formed from all-trans-retinal, such as lipofuscin or N-retinylidine-N-retinylethanolamine (A2E). In yet another embodiment slowing or halting the progression of RP is associated with correcting the folding of mutated opsins. In another embodiment, the administering is topical administration, local administration (e.g., intraocular or 20 periocular injection or implant) or systemic administration (e.g., oral, injection). In yet another embodiment, the light toxicity is related to an ophthalmic procedure (e.g., ophthalmic surgery). In still another embodiment, the administering occurs prior to, during, or after the ophthalmic surgery. 25 In one aspect, the invention provides a method of correcting mislocalized opsin within a photoreceptor cell, comprising contacting a mislocalized opsin protein with an opsin-binding agent that binds reversibly and/or non-covalently to said mislocalized opsin protein to promote the appropriate intracellular processing and transport of said opsin protein. 30 In various embodiments, the ophthalmic condition is any one or more of wet or dry form of macular degeneration, retinitis pigmentosa, a retinal or macular dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal 9 dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity, retinitis pigmentosa, normal vision loss related aging and normal loss of night vision related to aging. 5 In still another embodiment, the method further involves administering to a mammal, preferably a human being, an effective amount of at least one additional agent selected from the group consisting of a proteasomal inhibitor, an autophagy inhibitor, a lysosomal inhibitor, an inhibitor of protein transport 10 from the ER to the Golgi, an Hsp90 chaperone inhibitor, a heat shock response activator, a glycosidase inhibitor, and a histone deacetylase inhibitor. In yet another embodiment, the opsin binding ligand and the additional agent are administered simultaneously. 15 In still another embodiment, the opsin binding ligand and the additional agent are each incorporated into a composition that provides for their long term release. In another embodiment, the composition is part of a microsphere, nanosphere, nano emulsion or implant. In another embodiment, the composition further involves administering a mineral supplement, at least 20 one anti-inflammatory agent, such as a steroid (e.g., any one or more of cortisone, hydrocortisone, prednisone, prednisolone, methylprednisolone, triamcinolone, betamethasone, beclamethasone and dexamethasone), or at least one anti-oxidant, such as vitamin A, vitamin C and vitamin E. In various embodiments, the opsin binding ligand, the anti-inflammatory agent, and/or 25 the anti-oxidant are administered simultaneously. 30 10 BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows predicted hydration of the rod opsin retinal binding pocket as developed from a homology model of human rhodopsin based upon 5 the crystal structure of bovine rhodopsin. As a reference, the surface volume of 11-cis retinal is indicated by general outline and the structure of 11-cis retinal is indicated by bold black lines . Specific hydration sites are shown as circles where water molecules would be predicted to reside within the pocket in the absence of a ligand. Circles labeled with a "D" designate hydration sites 10 that are in very hydrophobic environments and thus upon displacement by a ligand are predicted to lower the energy of the ligand protein complex relative to the hydrated apoprotein. Circles labeled with a "R" designate hydration sites where the water molecule is forming stable hydrogen bonds with functional groups on the protein and thus signify coordinates within the 15 binding pocket where suitable hydrogen bonding functionality of the ligand should be incorporated to replace the hydrogen bonding interactions that are broken between the water molecule and the protein upon binding of the ligand. 20 Figure 2 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 gM of compound 6 during mutant protein production relative to pigment formation in the presence of vehicle, here dimethylsulfoxide (DMSO), alone. 25 Figure 3 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 gM of compound 13 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 30 Figure 4 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 M of compound 33 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone.

11 Figure 5 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 37 during mutant protein production relative to pigment 5 formation in the presence of vehicle (DMSO) alone. Figure 6 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 50 during mutant protein production relative to pigment 10 formation in the presence of vehicle (DMSO) alone. Figure 7 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 51 during mutant protein production relative to pigment 15 formation in the presence of vehicle (DMSO) alone. Figure 8 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 52 during mutant protein production relative to pigment 20 formation in the presence of vehicle (DMSO) alone. Figure 9 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 53 during mutant protein production relative to pigment 25 formation in the presence of vehicle (DMSO) alone. Figure 10 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 55 during mutant protein production relative to pigment 30 formation in the presence of vehicle (DMSO) alone.

12 Figure 11 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 57 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 5 Figure 12 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 63 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 10 Figure 13 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 71 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 15 Figure 14 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 73 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 20 Figure 15 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 80 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 25 Figure 16 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 pM of compound 105 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 30 Figure 17 shows the increase in regeneration of 500 nm absorbing pigment upon treatment with retinal from P23H opsin that was treated with 20 13 pM of compound 106 during mutant protein production relative to pigment formation in the presence of vehicle (DMSO) alone. 5 DEFINITIONS As used throughout the disclosure, the following terms, unless 10 otherwise indicated, shall be understood to have the following meanings. By "mislocalization" of a photoreceptor cell visual pigment protein (for example, opsin, especially human opsin) is meant that the synthesized protein is not found at the normal or appropriate cellular location. 15 "Pharmacologic chaperones" refer to small molecular weight chemical compounds that interact with a protein (usually with a mis-folded, or un-folded protein) in such a way as to alter the folding or confirmation of said protein. Such an interaction can have diverse consequences on the cellular fate of 20 the protein, including but not limited to leading to increased stability and increased levels of functional protein, increased stability and increased levels of non-functional protein, or decreased stability and decreased levels of functional or non-functional protein. 25 "Productive chaperone" refers to a pharmacologic chaperone that when interacting with a protein leads to an increased level of functional protein. "Counterproductive, shipwreck or destructive chaperone" refers to a pharmacologic chaperone that interacts with a protein (usually with a mis 30 folded, or un-folded protein) and this interaction leads to a decreased stability and/or decreased levels of functional or non-functional protein. By "proteasomal inhibitor" is meant a compound that reduces a proteasomal activity, such as the degradation of a ubiquinated protein.

14 By "autophagy inhibitor" is meant a compound that reduces the degradation of a cellular component by a cell in which the component is located. 5 By "lysosomal inhibitor" is meant a compound that reduces the intracellular digestion of macromolecules by a lysosome. In one embodiment, a lysosomal inhibitor decreases the proteolytic activity of a lysosome. 10 By "Inhibitor of ER-Golgi protein transport" is meant a compound that reduces the transport of a protein from the ER (endoplasmic reticulum) to the Golgi, or from the Golgi to the ER. By "HSP90 chaperone inhibitor" is meant a compound that reduces the 15 chaperone activity of heat shock protein 90 (HSP90). In one embodiment, the inhibitor alters protein binding to an HSP90 ATP/ADP pocket. By "heat shock response activator" is meant a compound that increases the chaperone activity or expression of a heat shock pathway 20 component. Heat shock pathway components include, but are not limited to, HSP100, HSP90, HSP70, HASP60, HSP40 and small HSP family members. By "glycosidase inhibitor" is meant a compound that reduces the activity of an enzyme that cleaves a glycosidic bond. 25 By "histone deacetylase inhibitor" is meant a compound that reduces the activity of an enzyme that deacetylates a histone. By "reduces" or "increases" is meant a negative or positive alteration, 30 respectively. In particular embodiments, the alteration is by at least about 10%, 25%, 50%, 75%, or 100% of the initial level of the protein produced in the absence of the opsin binding ligand.

15 As used herein, the term "wild-type conformation" refers to the three dimensional conformation or shape of a protein that is free of mutations to its amino acid sequence. For opsin, this means a protein free from mutations that cause misfiring, such as the mutation designated P23H (meaning that a 5 proline is replaced by a histidine at residue 23 starting from the N-terminus). Opsin in a "wild-type conformation" is capable of opsin biological function, including but not limited to, retinoid binding, visual cycle function, and insertion into a photoreceptor membrane. 10 By "agent" is meant a small compound (also called a "compound"), polypeptide, polynucleotide, or fragment thereof. The terms compound and agent are used interchangeably unless specifically stated otherwise herein for a particular agent or compound. 15 By "correcting the conformation" of a protein is meant inducing the protein to assume a conformation having at least one biological activity associated with a wild-type protein. By "misfolded opsin protein" is meant a protein whose tertiary structure 20 differs from the conformation of a wild-type protein, such that the misfolded protein lacks one or more biological activities associated with the wild-type protein. By "selectively binds" is meant a compound that recognizes and binds 25 a polypeptide of the invention, such as opsin, but which does not substantially recognize and bind other molecules, especially non-opsin polypeptides, in a sample, for example, a biological sample. By "effective amount" or "therapeutically effective amount" is meant a 30 level of an agent sufficient to exert a physiological effect on a cell, tissue, or organ or a patient. As used herein, it is the amount sufficient to effect the methods of the invention to achieve the desired result.

16 By "pharmacological chaperone" is meant a molecule that upon contacting a mutant protein is able to facilitate/stabilize the proper folding of the protein such that it acts and functions much more like wild type protein than would be the case in the absence of the molecule. 5 By "control" is meant a reference condition. For example, where a cell contacted with an agent of the invention is compared to a corresponding cell not contacted with the agent, the latter is the "control" or "control" cell. 10 By "treat" is meant decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease, preferably an ocular disease, such as RP, AMD and/or light toxicity. By "prevent" is meant reduce the risk that a subject will develop a 15 condition, disease, or disorder, preferably an ocular disease, such as RP, AMD and/or light toxicity. By "competes for binding" is meant that a compound of the invention and an endogenous ligand are incapable of binding to a target at the same 20 time. Assays to measure competitive binding are known in the art, and include, measuring a dose dependent inhibition in binding of a compound of the invention and an endogenous ligand by measuring t 1

/

2 , for example. A "pharmaceutically acceptable salt" is a salt formed from an acid or a 25 basic group of one of the compounds of the invention. Illustrative salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbatc, succinate, maleate, gentisinate, fumarate, gluconate, 30 glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesuifonate, and pamoate (i.e., 1,1'-methytene-bis-(2-hydroxy-3-naphthoate)) salts.

17 The term "pharmaceutically acceptable salt" also refers to a salt prepared from a compound of the invention having an acidic functional group, such as a carboxylic acid functional group, and a pharmaceutically acceptable inorganic or organic base. Suitable bases include, but are not limited to, 5 hydroxides of alkali metals such as sodium, potassium, and lithium; hydroxides of alkaline earth metal such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia, and organic amines, such as unsubstituted or hydroxy-substituted mono-, di-, or trialkylamines; dicyclohexylamine; tributyl amine; pyridine; N-methyl-N 10 ethylamine; diethylamine; triethylamine; mono-, bis-, or tris-(2-hydroxy-lower alkylamines), such as mono-, bis-, or tris-(2-hydroxyethyl)- amine, 2-hydroxy tert-butylamine, or tris-(hydroxymethyl)methylamine, N, N,-di-lower alkyl-N (hydroxy lower alkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl)-amine, or tri-(2-hydroxyethyl)amine; N-methyl-D-glucamine; and amino acids such as 15 arginine, lysine, and the like. The term "pharmaceutically acceptable salt" also refers to a salt prepared from a compound disclosed herein, e.g., a salt of a compound of Example 1 , having a basic functional group, such as an amino functional 20 group, and a pharmaceutically acceptable inorganic or organic acid. Suitable acids include, but are not limited to, hydrogen sulfate, citric acid, acetic acid, oxalic acid, hydrochloric acid, hydrogen bromide, hydrogen iodide, nitric acid, phosphoric acid, isonicotinic acid, lactic acid, salicylic acid, tartaric acid, ascorbic acid, succinic acid, maleic acid, besylic acid, fumaric acid, 25 gluconic acid, glucaronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. The term "pharniaceutically-acceptable excipient" as used herein 30 means one or more compatible solid or liquid tiller, diluents or encapsulating substances that are suitable for administration into a human. The term "excipient" includes an inert substance added to a pharmacological composition to further facilitate administration of a compound. Examples of 18 excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols. 5 The term "carrier" denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate administration. The term "parenteral" includes subcutaneous, intrathecal, intravenous, 10 intramuscular, intraperitoncal, or infusion. The term "visual cycle product" refers to a chemical entity produced as a natural product of one or more reactions of the visual cycle (the reactive cycle whereby opsin protein binds 11-cis-retinal to form rhodopsin, which 15 accepts a light impulse to convert 11-cis-retinal to all trans-retinal, which is then released from the molecule to regenerate opsin protein with subsequent binding of a new 11-cis-retinal to regenerate rhodopsin). Such visual cycle products include, but are not limited to, all-trans-retinal, lipofuscin and A2E. 20 The term "light toxicity" refers to any condition affecting vision that is associated with, related to, or caused by the production and/or accumulation of visual cycle products. Visual cycle products include, but are not limited to, all-trans-retinal, lipofuscin or A2E. In one particular embodiment, light toxicity is related to exposure of the eye to large amounts of light or to very high light 25 intensity, occurring, for example, during a surgical procedure on the retina. The term "opsin" refers to an opsin protein, preferably a mammalian opsin protein, most preferably a human opsin protein. In one embodiment, the opsin protein is in the wild-type (i.e., physiologically active) conformation. 30 One method of assaying for physiological activity is assaying the ability of opsin to bind 11-cis-retinal and form active rhodopsin. A mutant opsin, such as the P23H mutant, that is ordinarily misfolded has a reduced ability to bind 11-cis-retinal, and therefore forms little or no rhodopsin. Where the 19 conformation of the mutant opsin has been corrected (for example, by binding to a pharmacological chaperone), the opsin is correctly inserted into the rod cell membrane so that its conformation is the same, or substantially the same, as that of a non-mutant opsin. This allows the mutant opsin to bind 11-cis 5 retinal to form active rhodopsin. Therefore, the methods of the invention operate to reduce the formation of visual cycle products. "Alkyl" refers to an unbroken non-cyclic chain of carbon atoms that may be substituted with other chemical groups. It may also be branched or 10 unbranched, substituted or unsubstituted. "Lower alkyl" refers to a branched or straight chain acyclic alkyl group comprising one to ten carbon atoms, preferably one to eight carbon atoms, more preferably one to six carbon atoms. Exemplary lower alkyl groups 15 include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, neopentyl, iso-amyl, hexyl, and octyl. All alkyl, alkenyl or alkynyl groups disclosed herein may be substituted with one or more of the following: lower alkyl, hydroxy, ester, amidyl, oxo, 20 carboxyl, carboxamido, halo, cyano, nitrate, nitrite, thionitrate, thionitrite sulfhydryl and amino groups (as elsewhere defined herein). "Haloalkyl" refers to a lower alkyl group, an alkenyl group, an alkynyl group, a bridged cycloalkyl group, a cycloalkyl group or a heterocyclic ring, as 25 defined herein, to which is appended one or more halogens, as defined herein. Exemplary haloalkyl groups include trifluoromethyl, chloromethyl, 2 bromobutyl and 1-bromo-2-chloro-pentyl. "Alkenyl" refers to a branched or straight chain C2-C10 hydrocarbon 30 (preferably a C2-C8 hydrocarbon, more preferably a C2-C hydrocarbon) that can comprise one or more carbon-carbon double bonds. Exemplary alkenyl groups include propylenyl, buten-1-yl, isobutenyl, penten-1 -yl, 2,2 methylbuten-1-yl, 3-methylbuten-1-yi, hexan-1-yl, hepten-1-yl and octen-1-yl.

20 "Lower alkenyl" refers to a branched or straight chain C 2

-C

4 hydrocarbon that can comprise one or two carbon-carbon double bonds. 5 "Substituted alkenyl" refers to a branched or straight chain C 2 -C1O hydrocarbon (preferably a C 2

-C

8 hydrocarbon, more preferably a C 2

-C

6 hydrocarbon) which can comprise one or more carbon-carbon double bonds, wherein one or more of the hydrogen atoms have been replaced with one or more R100 groups, wherein each R 100 is independently a hydroxy, an oxo, a 10 carboxyl, a carboxamido, a halo, a cyano or an amino group, as defined herein. "Alkynyl" refers to an unsaturated acyclic C 2

-C

10 hydrocarbon (preferably a C 2

-C

8 hydrocarbon, more preferably a C 2

-C

6 hydrocarbon) that 15 can comprise one or more carbon-carbon triple bonds. Exemplary alkynyl groups include ethynyl, propynyl, butyn-1-yl, butyn-2-yl, pentyl-1-yl, pentyl-2 yl, 3-methylbutyn-1-yl, hexyl-1-yl, hexyl-2-yl, hexyl-3-yl and 3,3-dimethyl butyn-1-yl. 20 "Lower alkynyl" refers to a branched or straight chain C 2

-C

4 hydrocarbon that can comprise one or two carbon-carbon triple bonds "Bridged cycloalkyl" refers to two or more cycloalkyl groups, heterocyclic groups, or a combination thereof fused via adjacent or non 25 adjacent atoms. Bridged cycloalkyl groups can be unsubstituted or substituted with one, two or three substituents independently selected from alkyl, alkoxy,- amino, alkylamino, dialkylamino, hydroxy, halo, carboxyl, alkylcarboxylic acid, aryl, amidyl, ester, alkylcarboxylic ester, carboxamido, alkylcarboxamido, oxo and nitro. Exemplary bridged cycloalkyl groups include 30 adamantyl, decahydronapthyl, quinuclidyl, 2,6-dioxabicyclo(3.3.0)octane, 7 oxabicyclo(2.2.1)heptyl and 8-azabicyclo(3,2,1)oct-2-enyl.

21 "Cycloalkyl" refers to a saturated or unsaturated cyclic hydrocarbon comprising from about 3 to about 10 carbon atoms. Cycloalkyl groups can be unsubstituted or substituted with one, two or three substituents independently selected from alkyl, alkoxy, amino, alkylamino, dialkylamino, arylamino, 5 diarylamino, alkylarylamino, aryl, amidyl, ester, hydroxy, halo, carboxyl, alkylcarboxylic acid, alkylcarboxylic ester, carboxamido, alkylcarboxamido, oxo, alkylsulfinyl, and nitro. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl and cyclohepta-1,3-dienyl. 10 "Heterocyclic ring or group" refers to a saturated or unsaturated cyclic or polycyclic hydrocarbon group having about 2 to about 12 carbon atoms where 1 to about 4 carbon atoms are replaced by one or more nitrogen, oxygen and/or sulfur atoms. Sulfur may be in the thio, sulfinyl or sulfonyl oxidation state. The heterocyclic ring or group can be fused to an aromatic 15 hydrocarbon group. Heterocyclic groups can be unsubstituted or substituted with one, two or three substituents independently selected from alkyl, alkoxy, amino, alkylthio, aryloxy, arylthio, arylalkyl, hydroxy, oxo, thial, halo, carboxyl, carboxylic ester, alkylcarboxylic acid, alkylcarboxylic ester, aryl, arylcarboxylic acid, arylcarboxylic ester, amidyl, ester, alkylcarbonyl, arylcarbonyl, 20 alkylsulfinyl, carboxamido, alkylcarboxamido, arylcarboxamido, sulfonic acid, sulfonic ester, sulfonamide nitrate and nitro. Exemplary heterocyclic groups ,include pyrrolyl, furyl, thienyl, 3-pyrrolinyl,4,5,6-trihydro-2H-pyranyl, pyridinyl, 1,4-dihydropyridinyl, pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl, oxazolyl, thiazolyl, thieno[2,3-d]pyrimidine, 4,5,6,7-tetrahydrobenzo[b]thiophene, 25 imidazolyl, indolyl, thiophenyl, furanyl, tetrahydrofuranyl, tetrazolyl, pyrrolinyl, pyrrolindinyl, oxazolindinyl 1,3-dioxolanyl, imidazolinyl, imidazolindinyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3 triazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4H-pyranyl, piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl, pyrazinyl, piperazinyl, 1,3,5 30 triazinyl, 1,3,5-trithianyl, benzo(b)thiophenyl, benzimidazolyl, benzothiazolinyl, quinolinyl and 2,6-dioxabicyclo(3.3.0)octane.

22 "Heterocyclic compounds" refer to mono- and polycyclic compounds comprising at least one aryl or heterocyclic ring. "Aryl" refers to a monocyclic, bicyclic, carbocyclic or heterocyclic ring 5 system comprising one or two aromatic rings. Exemplary aryl groups include phenyl, pyridyl, napthyl, quinoyl, tetrahydronaphthyl, furanyl, indanyl, indenyl, indoyl. Aryl groups (including bicyclic aryl groups) can be unsubstituted or substituted with one, two or three substituents independently selected from alkyl, alkoxy, alkylthio, amino, alkylamino, dialkylamino, arylamino, 10 diarylamino, alkylarylamino, halo, cyano, alkylsulfinyl, hydroxy, carboxyl, carboxylic ester, alkylcarboxylic acid, alkylcarboxylic ester, aryl, arylcarboxylic acid, arylcarboxylic ester, alkylcarbonyl, arylcarbonyl, amidyl, ester, carboxamido, alkylcarboxamido, carbomyl, sulfonic acid, sulfonic ester, sulfonamido and nitro. Exemplary substituted aryl groups include 15 tetrafluorophenyl, pentafluorophenyl, sulfonamide, alkylsulfonyl and arylsulfonyl. "Cycloalkenyl" refers to an unsaturated cyclic C 3

-C

10 hydrocarbon (preferably a C 3

-C

8 hydrocarbon, more preferably a .C 3

-C

6 hydrocarbon), 20 which can comprise one or more carbon-carbon double bonds. "Alkylaryl" refers to an alkyl group, as defined herein, to which is appended an aryl group, as defined herein. Exemplary alkylaryl groups include benzyl, phenylethyl, hydroxybenzyl, fluorobenzyl and 25 fluorophenylethyl. "Arylalkyl" refers to an aryl radical, as defined herein, attached to an alkyl radical, as defined herein. Exemplary arylalkyl groups include benzyl, phenylethyl, 4-hydroxybenzyl, 3-fluorobenzyl and 2-fluorophenylethyl. 30 "Arylalkenyl" refers to an aryl radical, as defined herein, attached to an alkenyl radical, as defined herein. Exemplary arylalkenyl groups include styryl and propenylphenyl.

23 "Cycloalkylalkyl" refers to a cycloalkyl radical, as defined herein, attached to an alkyl radical, as defined herein. 5 "Cycloalkylalkoxy" refers to a cycloalkyl radical, as defined herein, attached to an alkoxy radical, as defined herein. "Cycloalkylalkylthio" refers to a cycloalkyl radical, as defined herein, attached to an alkylthio radical, as defined herein. 10 "Heterocyclicalkyl" refers to a heterocyclic ring radical, as defined herein, attached to an alkyl radical, as defined herein. "Arylheterocyclic ring" refers to a bi- or tricyclic ring comprised of an 15 aryl ring, as defined herein, appended via two adjacent carbon atoms of the aryl ring to a heterocyclic ring, as defined herein. Exemplary arylheterocyclic rings include dihydroindole and 1,2,3,4-tetra-hydroquinoline. "Alkylheterocyclic ring" refers to a heterocyclic ring radical, as defined 20 herein, attached to an alkyl radical, as defined herein. Exemplary alkylheterocyclic rings include 2-pyridylmethyl and 1-methylpiperidin-2-one-3 methyl. "Alkoxy" refers to R 5 oO-, wherein R 50 is an alkyl group, as defined 25 herein (preferably a lower alkyl group or a haloalkyl group, as defined herein). Exemplary alkoxy groups include methoxy, ethoxy, t-butoxy, cyclopentyloxy and trifluoromethoxy. "Aryloxy" refers to R 55 0-, wherein R 55 is an aryl group, as defined 30 herein. Exemplary arylkoxy groups include napthyloxy, quinolyloxy, isoquinolizinyloxy.

24 "Alkylthio" refers to R 50 S-, wherein R 50 is an alkyl group, as defined herein. "Lower alkylthio" refers to a lower alkyl group, as defined herein, 5 appended to a thio group, as defined herein. "Arylalkoxy" or "alkoxyaryl" refers to an alkoxy group, as defined herein, to which is appended an aryl group, as defined herein. Exemplary arylalkoxy groups include benzyloxy, phenylethoxy and chlorophenylethoxy. 10 "Arylalklythio" refers to an alkylthio group, as defined herein, to which is appended an aryl group, as defined herein. Exemplary arylalklythio groups include benzylthio, phenylethylthio and chlorophenylethylthio. 15 "Arylalkylthioalkyl" refers to an arylalkylthio group, as defined herein, to which is appended an alkyl group, as defined herein. Exemplary arylalklythioalkyl groups include benzylthiomethyl, phenylethylthiomethyl and chlorophenylethylthioethyl. 20 "Alkylthioalkyl" refers to an alkylthio group, as defined herein, to which is appended an alkyl group, as defined herein. Exemplary alkylthioalkyl groups include allylthiomethyl, ethylthiomethyl and trifluoroethylthiomethyl. "Alkoxyalkyl" refers to an alkoxy group, as defined herein, appended to 25 an alkyl group, as defined herein. Exemplary alkoxyalkyl groups include methoxymethyl, methoxyethyl and isopropoxymethyl. "Alkoxyhaloalkyl" refers to an alkoxy group, as defined herein, appended to a haloalkyl group, as defined herein. Exemplary alkoxyhaloalkyl 30 groups include 4- methoxy-2-chlorobutyl.

25 "Cycloalkoxy" refers to R 54 0-, wherein R 54 is a cycloalkyl group or a bridged cycloalkyl group, as defined herein. Exemplary cycloalkoxy groups include cyclopropyloxy, cyclopentyloxy and cyclohexyloxy. 5 "Cycloalkylthio" refers to R 54 S-, wherein R 54 is a cycloalkyl group or a bridged cycloalkyl group, as defined herein. Exemplary cycloalkylthio groups include cyclopropylthio, cyclopentylthio and cyclohexylthio. "Haloalkoxy" refers to an alkoxy group, as defined herein, in which one 10 or more of the hydrogen atoms on the alkoxy group are substituted with halogens, as defined herein. Exemplary haloalkoxy groups include 1,1,1 trichloroethoxy and 2-bromobutoxy. "Hydroxy" refers to -OH. 15 "Oxy" refers to -0-. "Oxo" refers to =0. 20 "Oxylate" refers to -0- R 77 + wherein R 77 is an organic or inorganic cation. "Thiol" refers to -SH. 25 "Thio" refers to -S-. "Oxime" refers to =N-OR 8 1 wherein R 8 1 is a hydrogen, an alkyl group, an aryl group, an alkylsulfonyl group, an arylsulfonyl group, a carboxylic ester, an alkylcarbonyl group, an arylcarbonyl group, a carboxamido group, an 30 alkoxyalkyl group or an alkoxyaryl group. "Hydrazone" refers to =N-N(R 81

)(R'

81 ) wherein R' 81 is independently selected from R 81 , and R 8 1 is as defined herein.

26 "Hydrazino" refers to H 2 N-N(H)-. "Organic cation" refers to a positively charged organic ion. Exemplary 5 organic cations include alkyl substituted ammonium cations. "Inorganic cation" refers to a positively charged metal ion. Exemplary inorganic cations include Group I metal cations such as for example, sodium, potassium, magnesium and calcium. 10 "Hydroxyalkyl" refers to a hydroxy group, as defined herein, appended to an alkyl group, as defined herein. "Nitrate" refers to -O-NO 2 i.e. oxidized nitrogen. 15 "Nitrite" refers to -O-NO i.e. oxidized nitrogen. "Nitro" refers to the group -NO 2 and "nitrosated" refers to compounds that have been substituted therewith. 20 "Nitroso" refers to the group -NO and "nitrosylated" refers to compounds that have been substituted therewith. "Nitrile" and "cyano" refer to -CN. 25 "Halogen" or "halo" refers to iodine (1), bromine (Br), chlorine (Cl), and/or fluorine (F). "Imine" refers to -C(=N-R 5 1 )- wherein R 5 1 is a hydrogen atom, an alkyl 30 group, an aryl group or an arylheterocyclic ring, as defined herein. "Amine" refers to any organic compound that contains at least one basic nitrogen atom.

27 "Amino" refers to -NH 2 , an alkylamino group, a dialkylamino group, an arylamino group, a diarylamino group, an alkylarylamino group or a heterocyclic ring, as defined herein. 5 "Alkylamino" refers to R 50 NH-, wherein R 50 is an alkyl group, as defined herein. Exemplary alkylamino groups include methylamino, ethylamino, butylamino, and cyclohexylamino. 10 "Arylamino" refers to R 55 NH-, wherein R 55 is an aryl group, as defined elsewhere herein. "Dialkylamino" refers to R 52

R

53 N-, wherein R 52 and R 5 3 are each independently an alkyl group, as defined herein. Exemplary dialkylamino 15 groups include dimethylamino, diethylamino and methyl propargylamino. "Diarylamino" refers to R 55

R

60 N-, wherein R 55 and R 60 are each independently an aryl group, as defined herein. 20 "Alkylarylamino" or "arylalkylamino" refers to R 52

R

55 N-, wherein R 5 2 is an alkyl group, as defined herein, and R 55 is an aryl group, as defined herein. "Alkylarylalkylamino " refers to R 52

R

79 N-, wherein R 5 2 is an alkyl group, as defined herein, and R 79 is an arylalkyl group, as defined herein. 25 "Alkylcycloalkylamino" refers to R 52

R

80 N-, wherein R 52 is an alkyl group, as defined herein, and R 80 is a cycloalkyl group, as defined herein. "Aminoalkyl" refers to an amino group, an alkylamino group, a 30 dialkylamino group, an arylamino group, a diarylamino group, an alkylarylamino group or a heterocyclic ring, as defined herein, to which is appended an alkyl group, as defined herein. Exemplary aminoalkyl groups 28 include dimethylaminopropyl, diphenylaminocyclopentyl and methylaminomethyl. "Aminoaryl" refers to an aryl group to which is appended an alkylamino 5 group, an arylamino group or an arylalkylamino group. Exemplary aminoaryl groups include anilino, N-methylanilino and N-benzylanilino. "Thio" refers to -S-. 10 "Sulfinyl" refers to -S(O)-. "Methanthial" refers to -C(S)-. "Thial" refers to =S. 15 "Sulfonyl" refers to -S(0)2~ "Sulfonic acid" refers to -S(0) 2 0R 7 6 , wherein R 76 is a hydrogen, an organic cation or an inorganic cation, as defined herein. 20 "Alkylsulfonic acid" refers to a sulfonic acid group, as defined herein, appended to an alkyl group, as defined herein. "Arylsulfonic acid" refers to a sulfonic acid group, as defined herein, 25 appended to an aryl group, as defined herein. "Sulfonic ester" refers to -S(0) 2 0R 58 , wherein R 58 is an alkyl group, an aryl group, or an aryl heterocyclic ring, as defined herein. 30 "Sulfonamido" refers to -S(0) 2

-N(R

1

)(R

5 7 ), wherein R 5 1 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group or an aryiheterocyclic ring, as defined herein, or R 5 1 and R 57 when taken together 29 are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein. "Alkylsulfonamido" refers to a sulfonamido group, as defined herein, 5 appended to an alkyl group, as defined herein. "Arylsulfonamido" refers to a sulfonamido group, as defined herein, appended to an aryl group, as defined herein. 10 "Alkylthio" refers to R 50 S-, wherein R 50 is an alkyl group, as defined herein (preferably a lower alkyl group, as defined herein). "Arylthio" refers to R 55 S-, wherein R 55 is an aryl group, as defined herein. 15 "Arylalkylthio" refers to an aryl group, as defined herein, appended to an alkylthio group, as defined herein. "Alkylsulfinyl" refers to R 50 -S(O)-, wherein R 50 is an alkyl group, as 20 defined herein. "Alkylsulfonyl" refers to R 50 -S(0) 2 -, wherein R 50 is an alkyl group, as defined herein. 25 "Alkylsulfonyloxy" refers to R 50

-S(O)

2 -0-, wherein R 50 is an alkyl group, as defined herein. "Arylsulfinyl" refers to R 55 -S(O)-, wherein R 55 is an aryl group, as defined herein. 30 "Arylsulfonyl" refers to R 55

-S(O)

2 -, wherein R 55 is an aryl group, as defined herein.

30 "Arylsulfonyloxy" refers to R 55

-S(O)

2 -0-, wherein R 55 is an aryl group, as defined herein. "Amidyl" refers to R 51

C(O)N(R

57 )- wherein R 5 1 and R 57 are each 5 independently a hydrogen atom, an alkyl group, an aryl group or an arylheterocyclic ring, as defined herein. "Ester" refers to R 5 1C(O)R 8 2 - wherein R 5 1 is a hydrogen atom, an alkyl group, an aryl group or an arylheterocyclic ring, as defined herein and R 82 is 10 oxygen or sulfur. "Carbamoyl" refers to -O-C(O)N(R 5 1

)(R

57 ), wherein R 5 1 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group or an aryiheterocyclic ring, as defined herein, or R 5 1 and R 57 taken together are a 15 heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein. "Carboxyl" refers to -C(O)OR 76 , wherein R 76 is a hydrogen, an organic cation or an inorganic cation, as defined herein. 20 "Carbonyl" refers to -C(O)-. "Alkylcarbonyl" refers to R 52 -C(O)-, wherein R 5 2 is an alkyl group, as defined herein. 25 "Arylcarbonyl" refers to R 55 -C(O)-, wherein R 55 is an aryl group, as defined herein. "Arylalkylcarbonyl" refers to R 5 5

-R

52 -C(O)-, wherein R 55 is an aryl 30 group, as defined herein, and R 5 2 is an alkyl group, as defined herein. "Alkylarylcarbonyl" refers to R 52

-R

55 -C(O)-, wherein R 55 is an aryl group, as defined herein, and R 52 is an alkyl group, as defined herein.

31 "Heterocyclicalkylcarbonyl" refer to R 78 C(O)- wherein R 78 is a heterocyclicalkyl group, as defined herein. 5 "Carboxylic ester" refers to -C(O)OR 58 , wherein R 58 is an alkyl group, an aryl group or an aryl heterocyclic ring, as defined herein. "Alkylcarboxylic acid" and "alkylcarboxyl" refer to an alkyl group, as defined herein, appended to a carboxyl group, as defined herein. 10 "Alkylcarboxylic ester" refers to an alkyl group, as defined herein, appended to a carboxylic ester group, as defined herein. "Alkyl ester" refers to an alkyl group, as defined herein, appended to an 15 ester group, as defined herein. "Arylcarboxylic acid" refers to an aryl group, as defined herein, appended to a carboxyl group, as defined herein. 20 "Arylcarboxylic ester" and "arylcarboxyl" refer to an aryl group, as defined herein, appended to a carboxylic ester group, as defined herein. "Aryl ester" refers to an aryl group, as defined herein, appended to an ester group, as defined herein. 25 "Carboxamido" refers to -C(O)N(R 5 1

)(R

57 ), wherein R 5 1 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group or an aryiheterocyclic ring, as defined herein, or R 5 1 and R 57 when taken together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as 30 defined herein. "Alkylcarboxamido" refers to an alkyl group, as defined herein, appended to a carboxamido group, as defined herein.

32 "Arylcarboxamido" refers to an aryl group, as defined herein, appended to a carboxamido group, as defined herein. "Urea" refers to -N(R 59

)-C(O)N(R

5 1)(R 57 ) wherein R 51 , R 57 , and R 59 are 5 each independently a hydrogen atom, an alkyl group, an aryl group or an aryiheterocyclic ring, as defined herein, or R 51 and R 57 taken together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein. 10 "Phosphoryl" refers to -P(R 70

)(R

71

)(R

72 ), wherein R 70 is a lone pair of electrons, thial or oxo, and R 7 1 and R 7 2 are each independently a covalent bond, a hydrogen, a lower alkyl, an alkoxy, an alkylamino, a hydroxy, an oxy or an aryl, as defined herein. 15 "Phosphoric acid" refers to -P(O)(OR 1 )OH wherein R 51 is a hydrogen atom, an alkyl group, an aryl group or an arylheterocyclic ring, as defined herein. "Phosphinic acid" refers to -P(O)(R 5 1 )OH wherein R 5 1 is a hydrogen 20 atom, an alkyl group, an aryl group or an aryiheterocyclic ring, as defined herein. "Silyl" refers to -Si(R 73

)(R

74 )(R7s), wherein R 73 , R 74 and R 75 are each independently a covalent bond, a lower alkyl, an alkoxy, an aryl or an 25 arylalkoxy, as defined herein. "Organic acid" refers to compound having at least one carbon atom and one or more functional groups capable of releasing a proton to a basic group. The organic acid preferably contains a carboxyl, a sulfonic acid or a 30 phosphoric acid moeity. Exemplary organic acids include acetic acid, benzoic acid, citric acid, camphorsulfonic acid, methanesulfonic acid, taurocholic acid, chlordronic acid, glyphosphate and medronic acid.

33 "Inorganic acid" refers to a compound that does not contain at least one carbon atom and is capable of releasing a proton to a basic group. Exemplary inorganic acids include hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid. 5 "Organic base" refers to a carbon containing compound having one or more functional groups capable of accepting a proton from an acid group. The organic base preferably contains an amine group. Exemplary organic bases include triethylamine, benzyldiethylamine, dimethylethyl amine, imidazole, 10 pyridine and pipyridine. "Independently selected" groups are groups present in the same structure that need not all represent the same substitution. For example, where two substituents are represented as NORA and each RA is said to be 15 independently selected from H, methyl, ethyl, etc., this means that where one RA is methyl, the other RA may be methyl but could be H or ethyl (or any other recited substitution). Some of the compounds for use in the methods of the present 20 invention may contain one or more chiral centers and therefore may exist in enantiomeric and diastereomeric forms. The scope of the present invention is intended to cover use of all isomers per se, as well as mixtures of cis and trans isomers, mixtures of diastereomers and racemic mixtures of enantiomers (optical isomers) as well. Further, it is possible using well known 25 techniques to separate the various forms, and some embodiments of the invention may feature purified or enriched species of a given enantiomer or diastereomer. A "pharmacological composition" refers to a mixture of one or more of 30 the compounds described herein, or pharmaceutically acceptable salts thereof, with other chemical components, such as pharmaceutically acceptable carriers and/or excipients. The purpose of a pharmacological composition is to facilitate administration of a compound to an organism.

34 The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject agent from one organ, or 5 portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato 10 starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol 15 and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. A physiologically 20 acceptable carrier should not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. A "solvate" is a complex formed by the combination of a solute (e.g., 25 a metalloprotease inhibitor) and a solvent (e.g., water). See J. Honig et al., The Van Nostrand Chemist's Dictionary, p. 650 (1953). The terms "optical isomer", "geometric isomer" (e.g., a cis and/or trans isomer), "stereoisomer", and "diastereomer" have the accepted 30 meanings (see, e.g., Hawley's Condensed Chemical Dictionary, 11th Ed.). The illustration of specific protected forms and other derivatives of the compounds of the instant invention is not intended to be limiting. The 35 application of other useful protecting groups, salt forms, prodrugs etc., is within the ability of the skilled artisan. A "prodrug" is a form of a drug that must undergo chemical conversion 5 by metabolic processes before becoming an active, or fully active, pharmacological agent. A prodrug is not active, or is less active, in its ingested or absorbed or otherwise administered form. For example, a prodrug may be broken down by bacteria in the digestive system into products, at least one of which will become active as a drug. Alternatively, it may be 10 administered systemically, such as by intravenous injection, and subsequently be metabolized into one or more active molecules. 15 DETAILED DESCRIPTION OF THE INVENTION In accordance with the present invention, it has been found that certain small molecule ligands are capable of reversibly binding non-covalently to the opsin protein and inhibiting the binding of 11-cis-retinal, to an opsin retinal 20 binding pocket. Such interference with retinal binding reduces the formation of visual cycle products, such as all-trans-retinal, and thereby inhibits the production of compounds such as lipofuscin and A2E with resulting reduced risk and occurrence of toxicity that can result from accumulation of these substances. Such compounds, acting as pharmacologic chaperones, are also 25 able to facilitate the proper folding and trafficking of mutant opsins associated with RP. Additionally, by inhibiting 11-cis-retinal binding and rhodopsin formation, the excessive stimulation and resulting activation of rhodopsin caused by exposure of the retina to bright light especially during retinal surgery reduces photocell death. 30 Certain synthetic retinoids (compounds structurally related to retinol (Vitamin A alcohol)) have been reported to bind to opsin. In the embodiments of the present invention, non-retinoid small molecules (compounds having a 36 molecular weight less than about 1000 daltons, less than 800, less than 600, less than 500, less than 400, or less than about 300 daltons) have been found to bind to opsin. 5 The invention features compositions and methods that are useful for reducing formation of visual cycle products and toxicity associated with the accumulation of such products in vivo, reducing the probability of apoptotic events associated with excessive rhodopsin activation as well as preventing rod cell death due to aberrant processing and trafficking of mutant opsin 10 proteins associated with RP. Mislocalization of photoreceptor cell visual pigment proteins (opsins) can occur in various ocular diseases, and also with normal aging. In such cases the accumulation of mislocalized opsin leads to the decline in viability of 15 photoreceptor cells. With time this mislocalized opsin accumulation leads to rod and cone cell death, retinal degeneration, and loss of vision. In one aspect, the invention provides a method of correcting mislocalized opsin within a photoreceptor cell, comprising contacting a 20 mislocalized opsin protein with an opsin-binding agent that binds reversibly and/or non-covalently to said mislocalized opsin protein, thereby promoting correct intracellular processing and transport of said opsin protein. Such opsin-binding agent is referred to as a "Productive Chaperone." 25 Such correction of mislocalization reduces photoreceptor cell stress, preventing photoreceptor cell decline in viability and death in various diseases of vision loss, and in normal age-related decline in dim-light and peripheral rod-mediated vision, central cone-mediated vision, and loss of night vision. 30 In another aspect of the invention, the opsin-binding agent promotes the degradation of the mislocalized opsin protein. This type of opsin-binding agent is referred to as a "Counterproductive", Shipwreck", or "Destructive Chaperone." 37 Enhancing the degradation of the mislocalized opsin by such an agent reduces the amount of mislocalized protein, thereby relieving photoreceptor cell stress, preventing decline in viability and death of photoreceptor cells in diseases of vision loss, as well as in normal age-related decline in dim-light 5 and peripheral rod-mediated vision, central cone-mediated vision, and loss of night vision. In embodiments of the foregoing, the ophthalmic condition is one or more of wet or dry form of macular degeneration, retinitis pigmentosa, a 10 retinal or macular dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity, retinitis pigmentosa, normal vision loss related aging and normal loss of night vision related to aging. 15 Opsin, the GPCR (G-protein coupled receptor) responsible for vision, readily regenerates with 11 -cis-retinal to form the visual pigment rhodopsin. The pigment is generated by formation of a protonated Schiff base between the aldehyde group of 1 1-cis-retinal and the s-amino group of L-lysine in opsin 20 (Matsumoto and Yoshizawa, Nature 1975 Dec 11;258(5535):523-6). Thus, the present invention provides compositions and methods of use of small molecule compounds that bind to wild type and mutant opsins and compete with, or other wise prevent, 1 1-cis-retinal from combining with opsin 25 to form rhodopsin and thereby inhibit formation of 1 1-cis-retinal and other visual cycle products. Binding to this site may be predicted by the efficiency upon which the ligand is able to displace and/or replace the waters in the various hydration 30 sites in the 11-cis retinal binding pocket as defined by the water map technology. Hydration sites labeled with an "R" (Figure 1 shows hydration sites as circles or spheres) that are occupied by waters that are predicted to have hydrogen bonding interactions with the protein. Thus, ligands that 38 displace these waters will ideally have functionality suitably oriented when the ligand binds to replace those hydrogen bonds that are broken in the process of the compound occupying the binding pocket. 5 In accordance with the present invention, ligand binding potency is enhanced by compounds that efficiently displace highly unstable waters from the opsin binding pocket. Occupation of the pocket by a pharmacologic chaperone creates interactions between the ligand and the protein which induce the proper folding and/or stabilization of the native 3-dimentional 10 conformation of the protein that leads to it being properly processed and trafficked to its proper location in the cell membrane. Alternatively, hydration sites labeled with a "D" (Figure 1) locate waters that are in hydrophobic environments and therefore it is optimal for the binding 15 compound to displace all of these waters with nonpolar substituents that compliment the hydrophobic environment of the protein. Thus, displacing waters in hydrophobic enviromments while replacing the hydrogen bonds of waters in hydration sites redicted to have hydrogen bonding interactions with the protein with functionality on the ligand that can act as water mimetics 20 when these waters are displaced leads to optimal potency and efficacy. Alternatively, displacing waters in hydration sites abeled with a "D" in Figure 1 and leaving those waters in hydration sites labeled with an "R: (shown in Figure 1) unperturbed such that their environment with the ligand bound does not adversely affect the intrinsic stability of these waters in the pocket in the 25 absence of ligand occupation leads to potent and efficacious compounds. The hydration sites are predicted locations of waters in the absence of a ligand based on the hydration map. Binding of a ligand of the invention may follow one of four possible mechanisms: (i) displacing a water occupying a hydration site, (ii) replacing a hydrogen bond between protein and a water in a hydration 30 site by a functionality of the ligand, (iii) binding of a ligand and leaving a water in the hydration site intact, and (iv) forming an extended hydrogen bonding network with the water in a hydration site while not displacing it.

39 In one embodiment, the invention provides opsin binding ligands of Formula (I) and pharmaceutically acceptable salts thereof: 5 A-B-Q-V Formula I wherein A is: 10 1)

R

1

R

2 Rd Rc R 3 Ra Rb 2)

R

1

R

2 Rd R24 Rc R 3 Ra Rb 15 3) R4 R5

R

T ; 4) N R8 40 5) R12 Rl Rl 6) NH 0 5 7) R14

R

16 8) R12 '. T R5 9)

R

17

R

18 10 T 10) RT9 R20/T 41 11) E 12) I'LN R21 5 13) 0 R T 14) R R 2 R93 Re; or 10 15) RI R 2 R9 B is: 1) -(CH 2 )n-; 42 2) -CH=CH-; 3) -CH 2

-N(R

2 2 )_; 4) -CH 2 -0 5) -C(O)-CH 2

-C(O)

5 6) N-E ;or 7) -C(O)NR 2 2 _ wherein n = 0, 1 or 2 and 10 E is: 1) -N(R 2 2 )-; or 2) oxygen; 15 Q is: 1) -C(O)-; 2) -(CH 2 )a 3) -S(0 2 )-; or 4) -CH 2

-C(O)

20 wherein a is 1 or 2; V is: 1) NR 2 1

R

22 ; 25 2)

R

25 R2 ) a -N Y

R

23

X

43 3) Y 4) ---N R26 0 5 5) N Y R22 ;or 6) R22 SN Q

R

23 wherein b is 1 or 2 and a is 1 or 2; 10 Y is: 1) NR 22 ; 2) N-Q-U; 3) CR 22

R

23 ; 15 4) oxygen; 5) S(O)n; 6) N-C(S)-NR 22

R

23 ; 7) N-(C=N-CN)-NR 22

R

23 8) N-(C=N-SO 2

CH

3

)-NR

22

R

23 ; 20 9) C=NOR 22 ; 10) C=N-NR 22

R

23 ;or 11) C-Q-U; and n is 0, 1 or 2; 25 U is: 1) NR 22

R

23

;

44 2) lower alkyl; 3) haloalkyl; 4) alkoxy; 5) OR 22 ; or 5 6) hydrogen; X is: 1) hydrogen 2) alkyl; or 10 3) -C=CR 9 ;

R

1 and R 2 are independently: 1) -CH 3 ; or 2) -CH 2

CH

3 ; 15 R3 is: 1) hydrogen; 2) -CH 3 ; or 3) -CH 2

CH

3 ; 20 Ra, and Rb, are each independently: 1) hydrogen; 2) deutero; or 3) -CH 3 25 Re, and Rd, are each independently: 1) hydrogen; 2) alkoxy; 3) lower alkyl; or 30 4) alkenyl; R4 is: 1) -CH 3

;

45 2) -CF 3 ; 3) -C 2

H

5 ; or 4) -C 3

H

5 ; 5 R , R 6 and R 7 are each independently: 1) hydrogen; 2) lower alkyl; 3) halogen; 4) dialkylamine; 10 5) nitro; or 6) dialkylamine; Z is: 1) CR 3 ; 15 2) CH; or 3) nitrogen; R is: 1) -CH 2 -; or 20 2) -C(O)-;

R

9 , R 14 and R 16 are each independently: 1) hydrogen; or 2) -CH 3 ; 25 R10 is: 1) N-R 13 ; 2) sulfur; or 3) oxygen; 30 R" is: 1) =N-; or 2) =C(CH 3

)-;

46 R is: 1) lower alkyl; 2) alkoxy; or 3) haloalkyl; 5 R is: 1) phenyl; 2) lower alkyl; or 3) haloalkyl; 10 R" is: 1) hydrogen; or 2) -C(O)CH 3 ; 15 R 17 and R 18 together are: 1) -(CH 2

)

4 -; or 2) -CH=CH-CH=CH

R

19 and R 20 together are: 20 1) -CH 2

-C(CH

3

)

2

-CH

2 -C(O)-; or 2) -CH=CH-CH=CH-; R21 is: 1) hydrogen; 25 2) -C(O)CH 3 ; 3) -CH 3 ; or 4) -CH 2

CH

3 ;

R

2 2 and R 2 3 are each independently: 30 1) hydrogen; or 2 lower alkyl;

R

24 and R 25 are each independently: 47 1) hydrogen; or 2) -CH 3 ;

R

26 is: 5 1) NR 22

R

23 : or 2) alkoxy; And wherein R 1 and R 2 taken together or Ra and Rb taken together 10 along with the carbon to which they are attached can form cyclopropyl;

R

24 and R 25 taken together along with the two carbons to which they are attached can form cyclopropyl: 15 R 24 and R 25 taken together can form oxo; And wherein T is: 1) oxygen; 20 2) -N(R 1 6 )-; or 3) sulfur; E is: 1) oxygen; 25 2) -N(R 1 6 )-; 3) sulfur; or 4) -C(O)-. In its broadest embodiments, R 1 , R 2 and R 3 are each independently 30 lower alkyl. In preferred embodiments, the compound has the structure of Formula I wherein V is: 48

R

25 R2 1#\a -N Y R23 X and wherein a and b are each independently 1 or 2, more preferably wherein at least one of a or b is 1, most preferably wherein both a and b are 1, X is 5 hydrogen, R 23 , R 24 and R 25 are hydrogen, Y is C-C(O)NR 22

R

23 or N

C(O)NR

22

R

23 and R 22 and R 23 are both hydrogen. In preferred examples of the invention, the compound has the structure of Formula I wherein A is: 10 RI R 2 Rd Rc R3 Ra Rb In preferred embodiments thereof, one or more of R 1 and R 2 is methyl, more preferably both are methyl, and R 3 is a hydrogen or a methyl 15 group. In other specific embodiments, Ra and Rb are independently hydrogen, deutero or methyl, preferably hydrogen or methyl, Re and Rd are preferably hydrogen or lower alkyl, most preferably hydrogen or methyl. In another preferred example, the compound has the structure of 20 Formula I wherein A is: RI R 2 Rd

R

24 Rc*R3 Ra Rb 49 In preferred embodiments thereof, R', R 2 and R 3 are each methyl, and

R

2 4 is a methyl or hydrogen, preferably a hydrogen. In other specific embodiments, Ra and Rb are independently hydrogen, 5 deutero or methyl, preferably hydrogen or methyl, Re and Rd are hydrogen lower alkyl, alkoxy or alkoxymethyl, more preferably hydrogen, alkoxy or lower alkyl, most preferably hydrogen. In another preferred example, the compound has Formula I wherein B 10 is -CH=CH-, -CH 2

-CH

2 - or -CH 2

-N(R

2 2 )-, preferably -CH=CH or -CH 2

-CH

2 -, and most preferably -CH=CH-. In another preferred example, the compound has Formula I wherein Q is -C(O)- or -CH 2 - , most preferably -C(O)-. 15 In another preferred example, the compound has Formula I wherein X is hydrogen, lower alkyl or -CaCR 9 , more preferably hydrogen or -C=CR 9 wherein R 9 is hydrogen or methyl. 20 In another preferred example, the compound has Formula I wherein Y is oxygen or N-C(O)-NR 2 2

R

2 3 , more preferably N-C(O)-NR 2 2

R

2 3 , most preferably N-C(O)-NR 22

R

23 wherein R 2 2 and R 2 3 are hydrogen. In another preferred example, the compound has Formula I wherein 25 R 24 and R 25 are each hydrogen. In another embodiment, the invention provides opsin binding ligands of Formula (1l) and pharmaceutically acceptable salts thereof: A-B-Q-NcjP W 30 Formula Il 50 wherein W is: 1) -OR 2 2 ; 2) -NR 22

R

2 3 3) -N(R 2 2

)-C(O)-NR

2 2

R

2 3 5 4) -O-C(O)-NR 22

R

23 ; 5) -N(R 2 2

)-C(S)-NR

22

R

2 3 6) -O-C(S)-NR 2 2

R

2 3 ; 7) -S-C(O)-NR 2 2

R

2 3 ; 8) -N(R 22

)-(C=N-CN)-NR

2 2

R

23 ; 10 9) -N(R 22

)-(C=N-SO

2 Me)-NR 22

R

23 ; or 10) -C(O)N(R 9

)N(R

4

)(R

16 ); In preferred examples, the compound has the structure of Formula Il wherein A is: 15

R

1

R

2 Rd Rc R 3 Ra Rb In preferred embodiments thereof, one or more of R 1 and R 2 is a methyl or ethyl group, preferably a methyl group, and R 3 is a hydrogen or a 20 methyl group. In other specific embodiments, Ra and Rb are independently hydrogen, deutero or methyl, preferably hydrogen or methyl, Re and Rd are hydrogen lower alkyl, alkoxy or alkoxymethyl, more preferably hydrogen, alkoxy or lower alkyl, most preferably hydrogen. 25 In another preferred example, the compound has Formula II wherein A is: 51

R

1

R

2 Rd R24 Ra Rb In preferred embodiments thereof, R 1 , R 2 , and R 3 is a ethyl or methyl group, more preferably a methyl group and R 24 is a methyl or hydrogen, 5 preferably a hydrogen. In other specific embodiments, Ra and Rb are independently hydrogen, deutero or methyl, preferably hydrogen or methyl, Rc and Rd are hydrogen lower alkyl, alkoxy or alkoxymethyl, more preferably hydrogen, alkoxy or lower alkyl, most preferably hydrogen. 10 In another preferred example, the compound has Formula II wherein B is -CH=CH-, -CH 2

-CH

2 - or -CH 2

-N(R

22 )-, preferably -CH=CH or -CH 2

-CH

2 -, and most preferably -CH=CH-. In another preferred example, the compound has Formula Il wherein Q 15 is -C(O)- or -CH 2 - most preferably -C(O)-. In another preferred example, the compound has Formula II wherein p is 0 or 1, most preferably 1. 20 In another preferred example, the compound has Formula 11 wherein W is -O-C(O)-NR 22

R

23 or -N(R 9

)-C(O)-NR

22

R

23 , more preferably -N(R 9

)-C(O)

NR

22

R

23 , most preferably -N(R 9

)-C(O)-NR

22

R

23 wherein each of R 9 , R 22 and

R

23 is hydrogen. 25 In specific embodiments the opsin binding compound of Formula I or Formula (11) is (wherein each compound number corresponds to the number of the example where it was prepared): (E)-3-(2,6,6-Trimethylcyclohex-1-enyl)acrylamide (Compound 2b); 52 (E)-N-Methyl-3-(2,6,6-trimethylcyclohex-1-enyl)acrylamide (Compound 3); (E)-N,N-Dimethyl-3-(2,6,6-trimethylcyclohex-1-enyl)acrylamide (Compound 4); (E)- -(Piperidin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 -one (Compound 5); 5 (E)-1 -Morpholino-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 -one (Compound 6); (E)-tert-Butyl 4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)-1,4-diazepane-1 carboxylate (Compound 7a); (E)-1 -(1,4-Diazepan-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 -one 10 (Compound 7b); (E)-1 -(4-Methyl- 1,4-diazepan-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2 en-1-one (Compound 7c); (E)-1-(4-Ethyl-1,4-diazepan-1-yl)-3-(2,6,6-trimethylcyclohex-1-enyl) prop-2-en 1-one (Compound 8); 15 (E)-1-(4-propyl-1,4-diazepan-1-yl)-3-(2,6,6-trimethylcyclohex-1-enyl) prop-2 en-i-one (Compound 9); (E)-1-(4-Acetyl-1,4-diazepan-1-yI)-3-(2,6,6-trimethylcyclohex-1-enyl) prop-2 en-i-one (Compound 10); (E)-1-(4-Propionyl-1,4-diazepan-1-yl)-3-(2,6,6-trimethylcyclohex-1-enyl)prop 20 2-en-1-one (Compound 11); (E)-1-(4-(2,2,2-Trifluoroacetyl)-1,4-diazepan-1-yi)-3-(2,6,6-trimethyl cyclohex 1-enyl)prop-2-en-1-one (Compound 12); (E)-4-(3-(2,6,6-Trimethylcyclohex-1-enyl)acryloyl)-1,4-diazepane-1 carboxamide (Compound 13); 25 (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)-1,4-diazepane-1 carboxamide (Compound 14); (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)-1,4-diazepane-1 carboxamide (Compound 15); (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)-1,4-diazepane-1 30 carboxamide (Compound 16); (E)-N-I sopropyl-4-(3-(2,6,6-trimethylcyclohex- 1 -enyl)acryloyl)-1,4-diazepane 1-carboxamide (Compound 17); 53 (E)-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)-1,4-diazepane-1 carboxylate (Compound 18); (E)-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)-1,4-diazepane-1 carboxylate (Compound 19); 5 (E)-1 -(4-(Methylsulfonyl)-1,4-diazepan-1 -yl)-3-(2,6,6-trimethylcyclo-hex-1 enyl)prop-2-en-1 -one (Compound 20); (E)-1 -(4-(Ethylsulfonyl)-1,4-diazepan-1 -yl)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1-one (Compound 21); (E)-1-(4-(Trifluoromethylsulfonyl)-1,4-diazepan-1-yl)-3-(2,6,6-tri 10 methylcyclohex-1-enyl)prop-2-en-1 -one (Compound 22); (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)-1,4-diazepane-1 carbothioamide (Compound 23); (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)-1,4-diazepane-1 carbothioamide (Compound 24); 15 (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)-1,4-diazepane-1 carbothioamide (Compound 25); (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)-1,4-diazepane 1-carbothioamide (Compound 26); (E)-1-(4-Methylpiperazin-1-yl)-3-(2,6,6-trimethylcyclohex-1-enyl)prop-2-en-1 20 one (Compound 27), (E)-tert-Butyl 4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl) piperazine-1 carboxylate (Compound 28a); (E)-i -(Piperazin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 -one (Compound 28b); 25 (E)-i -(4-Ethylpiperazin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 one (Compound 28c); (E)-i -(4-Propylpiperazin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 one (Compound 29), (E)-i -(4-Acetylpiperazin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 30 one (Compound 30); (E)-1-(4-Propionylpiperazin-1-yI)-3-(2,6,6-trimethylcyclohex-1-enyl) prop-2-en 1-one (Compound 31); 54 (E)-1 -(4-(2,2,2-Trifluoroacetyl) piperazin-1 -yl)-3-(2,6,6-trimethylcyclo-hex-1 enyl)prop-2-en-1-one (Compound 32); (E)-4-(3-(2,6,6-Trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 -carboxamide (Compound 33); 5 (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)piperazine-1 carboxamide (Compound 34); (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)piperazine-1 carboxamide (Compound 35); (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 10 carboxamide (Compound 36); (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 carboxamide (Compound 37); (E)-Methyl 4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl) piperazine-1 carboxylate (Compound 38); 15 (E)-Ethyl 4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl) piperazine-1 carboxylate (Compound 39); (E)-1 -(4-(Methylsulfonyl)piperazin-1 -yl)-3-( 2 ,6,6-trimethylcyclohex-1 -enyl)prop 2-en-i-one (Compound 40); (E)-1 -(4-(Ethylsulfonyl)piperazin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop 20 2-en-i-one (Compound 41); (E)-1-(4-(Trifluoromethylsulfonyl)piperazin-1-yl)-3-(2,6,6-trimethyl-cyclohex-1 enyl)prop-2-en-1-one (Compound 42); (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 carbothioamide (Compound 43); 25 (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 carbothioamide (Compound 44); (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 carbothioamide (Compound 45); (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 30 carbothioamide (Compound 46); (S,E)-1 -(3-Hydroxypyrrolidin-1 -yI)-3-(2,6,6-trimethylcyclohex-1 -enyl) prop-2 en-i-one (Compound 47); 55 (S,E)-1-(3-(2,6,6-Trimethylcyclohex-1-enyl)acryloyl)pyrrolidin-3-yI carbamate (Compound 48); (E)-tert-Butyl 1-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) pyrrolidin-3-yl carbamate (Compound 49a); 5 (E)-1 -(3-Aminopyrrolidin-1 -yI)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 one (Compound 49b); (E)-1 -(1 -(3-(2,6,6-Trimethylcyclohex-1 -enyl)acryloyl) pyrrolidin-3-yi) urea (Compound 50); 1-(Piperazin-1-yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)propan-1 -one (Compound 10 51a); 4-(3-(2,6,6-Trimethylcyclohex-1 -enyl)propanoyl)piperazine-1 -carboxamide (Compound 51b); (S, E)-2-Methyl-4-(3-(2,6,6-trimethylcyclohex- 1 -enyl)acryloyl)piperazine-1 carboxamide (Compound 52); 15 (R, E)-2-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 carboxamide (Compound 53);

(E)-N

2 -Methyl-4-(3-(2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl)piperazine-1,2-dicarboxamide (Compound 54); N-((2,6,6-Trimethylcyclohex-1 -en-1 -yl)methyl)piperazine-1,4 20 dicarboxamide (Compound 55); N'-Methyl-N 1 -((2,6,6-trimethylcyclohex-1 -en-1 -yl)methyl)piperazine 1,4-dicarboxamide (Compound 56); (R, E)-1 -(3-Hydroxypyrrolidin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -en-1 yl)prop-2-en-1-one (Compound 57a); 25 (RE)-1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl) pyrrolidin-3-yl carbamate (Compound 57b); (S,E)-1-(3-Aminopyrrolidin-1-yl)-3-(2,6,6-trimethylcyclohex-1-en-1 yl)prop-2-en-1-one (Compound 58b); (S,E)-1-(1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl)pyrrolidin-3 30 yl)urea (Compound 58c); (R,E)-1-(3-Aminopyrrolidin-1-yl)-3-(2,6,6-trimethylcyclohex-1-en-1 yl)prop-2-en-1-one (Compound 59b); (R,E)-1-(1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl) pyrrolidin-3- 56 yl)urea (Compound 59c); (E)-4-(3-(2,6,6-Trimethyl-3-oxocyclohex-1 -en-1 yl)acryloyl)piperazine-1-carboxamide (Compound 60); (E)-4-(3-(3,3-Difluoro-2,6,6-trimethylcyclohex-1-en-1 5 yl)acryloyl)piperazine-1-carboxamide (Compound 62); (E)-4-(3-(3,3-Dideutero-2,6,6-trimethylcyclohex-1 -en-1 yl)acryloyl) piperazine-1-carboxamide (Compound 63); (E)-1 -(1,1 -Dioxidothiomorpholino)-3-(2,6,6-trimethylcyclohex-1 eyl)prop-2-en-i-one (Compound 68); 10 (E)- -Thiomorpholino-3-(2,6,6-trimethylcyclohex-1 -en-1 yl)prop-2-en-i-one (Compound 69); (E)-1 -(4,4-Difluoropiperidin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 en-1 -yl)prop-2-en-1-one (Compound 70); (±)-4-((E)-3-((1,6-anti)-2,2,6-trimethylcyclohexyl) 15 acryloyl)piperazine-1-carboxamide (Compound 71); (-)-4-((E)-3-((i R, 6R)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxamide (Compound 72); (+)-4-((E)-3-((1 S,6S)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxamide (Compound 73); 20 (E)-i -Morpholino-3-((i R,6R)-2,2,6-trimethylcyclohexyl)prop-2 en-i-one (Compound 74); (E)-1 -Thiomorpholino-3-((i R,6R)-2,2,6-trimethylcyclohexyl) prop-2-en-i-one (Compound 75); (E)-4-(3-(2,5,6,6-tetramethylcyclohex-2-en-1 -yl)acryloyl) 25 piperazine-1-carboxamide (Compound 76); 4-((E)-3-((i R,6S)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 -carboxamide and 4-((E)-3-((i S,6R)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 carboxamide (Compound 77); (E)-4-(3-(2,6,6-trimethylcyclohex-2-en-1 -yl)acryloyl)piperazine 30 1-carboxamide (Compound 78); 4-(3-((i R,6S)-2,2,6-trimethyIcyclohexyl)propanoyl)piperazine-1 -carboxamide (Compound 80); 57 4-(3-((1 S,6R)-2,2,6-trimethylcyclohexyl)propanoyl)piperazine-1 -carboxamide (Compound 81): (E)-1 -Morpholino-3-((1 S,6S)-2,2,6-trimethylcyclohexyl)prop-2 en-1-one (Compound 83); 5 (E)-4-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl)piperazin 2-one (Compound 84); (E)-4-(3-(2,6,6-Trimethylcyclohex-1 -en-1 -yl)acryloyl)piperazine 1-carbaldehyde (Compound 88); (E)-1-(4-(2-hydroxyethyl)piperazin-1-yl)-3- (2,6,6 10 trimethylcyclohex- 1-en-1 -yl)prop-2-en-1 -one (Compound 89); (±)-3,5-cis-Dimethyl-4-((E)-3-(2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl) piperazine-1-carboxamide (Compound 90); (E)-4-(3-(2,2,6-trimethylbicyclo[4. 1.0]heptan-1 -yl)acryloyl) piperazine-1 carboxamide (Compound 91); 15 (±)-(E)-4-(3-(4-Methoxy-2,6,6-trimethylcyclohex-1-en-1 -yl) acryloyl) piperazine-1-carboxamide (Compound 93); (-)-((1 R,6S)-2,2,6-trimethylcyclohexyl)methyl 4-carbamoylpiperazine-1 carboxylate (Compound 94);

(-)-N

1 -Methyl-N-(((1 R,6S)-2,2,6-trimethylcyclohexyl)methyl) piperazine-1,4 20 dicarboxamide (Compound 95); N'-(((1 R,6S)-2,2,6-trimethylcyclohexyl)methyl)piperazine-1,4-dicarboxamide (Compound 96); 4-(((1 R,6S)-2,2,6-trimethylcyclohexanecarboxamido)methyl)piperidine-1 carboxamide (Compound 97); 25 (E)-2-(1 -(3-(2,6,6-trimethylcyclohex-1 -en-1 -yl)acryloyl)azetidin-3-yl) acetamide (Compound 98); (E)-3-(3-(2,6,6-Trimethylcyclohex-1 -en-1 -yl)acrylamido) azetidine-1 carboxamide (Compound 99); (E)-3-(2,6,6-Trimethylcyclohex-1 -en-1 -yl)-N-(2-ureidoethyl)acrylamide 30 (Compound 100); (E)-N-Methyl-N-(2-(1 -methylu reido)ethyl)-3-(2,6,6-trimethylcyclohex- 1-en-1 -yl) acrylamide (Compound 101); 58 (E)-4-(3-(2,2,6,6-Tetramethylcyclohexyl)acryloyl)piperazine-1 -carboxamide (Compound 102); (E)-1 -Morpholino-3-(2,2,6,6-tetramethylcyclohexyl)prop-2-en-1 -one (Compound 103); 5 N-((2,6,6-Trimethylcyclohex-1 -en-1 -yl)methyl)morpholine-4-carboxamide (Compound 104); (E)-4-(3-(2,6,6-Trimethylcyclohex-1 -en-1 -yl)acryloyl) piperazine-1 carbothioamide (Compound 105); (E)-2-Ethynyl-4-(3-(2,6,6-trimethylcyclohex-1 -en-1 -yl)acryloyl)piperazine- 1 10 carboxamide (Compound 106); (E)-1 -Morpholino-3-(3,3,6,6-tetramethylcyclohex-1 -enyl)prop-2-en-1 -one (Compound 107); (E)-4-(3-(3,3,6,6-Tetramethylcyclohex-1 -enyl)acryloyl) piperazine-1 carboxamide (Compound 108); 15 (E)-4-(3-(3,6,6-Trimethylcyclohex-1 -enyl)acryloyl)piperazine-1 -carboxamide (Compound 109); and (E)-1 -Morpholino-3-(3,6,6-trimethylcyclohex-1 -enyl)prop-2-en-1 -one (Compound 110); including all pharmaceutically acceptable salts, hydrates, or solvates thereof. 20 All compound names were derived using ChemBioDraw 11.0.1. Especially preferred examples of the compounds of the invention, and methods using said compounds, include compounds of Table 1, and are also 25 selected from one or more of the group consisting of compounds 6, 13, 14, 22, 33, 34, 37, 44, 45, 50, 51a, 51b, 52, 53, 55, 57, 60b, 63, 69, 71, 72, 73, 80, 84, 105, 106, 107, 108,109 and 110 including all pharmaceutically acceptable salts, solvates and hydrates thereof. 30 Another embodiment of the invention provides the opsin binding ligand metabolites of the opsin binding compounds. These metabolites, include but 59 are not limited to, degradation products, hydrolysis products, gluconoride adducts and the like, of the opsin binding compounds and pharmaceutically acceptable salts thereof, of the opsin compounds. 5 Another embodiment of the invention provides processes for making the novel compounds of the invention and to the intermediates useful in such processes. The reactions are performed in solvents appropriate to the reagents and materials used are suitable for the transformations being effected. It is understood by one skilled in the art of organic synthesis that the 10 functionality present in the molecule must be consistent with the chemical transformation proposed. This will, on occasion, necessitate judgment by the routineer as to the order of synthetic steps, protecting groups required, and deprotection conditions. Substituents on the starting materials may be incompatible with some of the reaction conditions required in some of the 15 methods described, but alternative methods and substituents compatible with the reaction conditions will be readily apparent to one skilled in the art. The use of sulfur, nitrogen and oxygen protecting groups is well known for protecting thiol, amino and alcohol groups against undesirable reactions during a synthetic procedure and many such protecting groups are known and 20 described by, for example, Greene and Wuts, Protective Groups in Organic Synthesis, Third Edition, John Wiley & Sons, New York (1999). Compounds of the invention that have one or more asymmetric carbon atoms may exist as the optically pure enantiomers, pure diastereomers, 25 mixtures of enantiomers, mixtures of diastereomers, racemic mixtures of enantiomers, diasteromeric racemates or mixtures of diastereomeric racemates. It is to be understood that the invention anticipates and includes within its scope all such isomers and mixtures thereof. 30 The chemical reactions described herein are generally disclosed in terms of their broadest application to the preparation of the compounds of this invention. Occasionally, the reactions may not be applicable as described to each compound included within the disclosed scope. The compounds for 60 which this occurs will be readily recognized by one skilled in the art. In all such cases, either the reactions can be successfully performed by conventional modifications known to one skilled in the art, e.g., by appropriate protection of interfering groups, by changing to alternative conventional 5 reagents, by routine modification of reaction conditions, or other reactions disclosed herein or otherwise conventional, will be applicable to the preparation of the corresponding compounds of this invention. In all preparative methods, all starting materials are known or readily prepared from known starting materials. 10 Methods of the invention The present invention provides a method of using compounds of the 15 Formula I and/or Formula 11 for reducing the formation of toxic visual cycle products, comprising contacting an opsin protein with small molecule ligands that reversibly bind to said opsin protein to inhibit 11-cis-retinal binding in said binding pocket, thereby reducing formation of toxic visual cycle products associated with wet or dry ARMD. and reducing photocell apoptosis 20 associatiated with excessive rhodopsin activation as a result of bright light stimulation. The present invention also provides a method of use of compounds of the Formula I and/or Formula 11 for treating, preventing or reducing the risk of 25 light toxicity in a mammal, comprising administering to a mammal, at risk of developing an ophthalmic condition that is related to the formation or accumulation of a visual cycle product or apoptotic photocell death. The present invention also provides a method of use of compounds of 30 the Formula I and/or Formula 11 for treating, preventing or reducing the risk of light toxicity in a mammal, comprising administering to a mammal, at risk of developing an ophthalmic condition that is related to the formation or accumulation of a visual cycle product or apoptotic photocell death, an 61 effective amount of a that small molecule ligand that reversibly binds (for example, at or near the retinal binding pocket) to an opsin protein present in the eye of said mammal, for example, to inhibit 11-cis-retinal binding in said binding pocket, thereby reducing light toxicity and photocell apoptosis. 5 The present invention also provides a method of use of compounds of the Formula I and/or Formula I for treating, preventing or reducing the risk of RP in a mammal, comprising administering to a mammal, at risk of RP related to the improper folding and trafficking of mutant opsins, an effective amount of 10 a that small molecule ligand that reversibly binds (for example, at or near the retinal binding pocket) to an opsin protein present in the eye of said mammal, for example, to inhibit 11-cis-retinal binding in said binding pocket, thereby reducing the vision loss caused by RP. 15 In specific examples of such methods, the small molecule ligand is selective for binding to opsin and/or the small molecule ligand binds to said opsin in the retinal binding pocket of said opsin protein and/or the small molecule ligand binds to said opsin protein so as to inhibit covalent binding of 11 -cis-retinal to said opsin protein when said 11 -cis-retinal is contacted with 20 said opsin protein when said small molecule ligand is present and/or the mammal is a human being. In one embodiment, light toxicity is related to an ophthalmic procedure, for example, ophthalmic surgery. Said agent may be administered prior to, 25 during or after said surgery (or at any one or more of those times). In specific embodiments of the methods of the invention, the native opsin protein is present in a cell, such as a rod cell, preferably, a mammalian and more preferably a human cell. In specific embodiments, the small 30 molecule ligands of the invention inhibit binding of 11-cis-retinal in the binding pocket of opsin and slow the visual cycle thereby reducing the formation of all trans-retinal, or a toxic visual cycle product formed from it, such as lipofuscin or N-retinylidene-N-retinylethanolamine (A2E). Alternatively, photocell 62 apoptosis as a result of excessive rhodopsin activation is reduced or prevented by inhibition of rhodopsin formation. Additionally, improper folding and trafficking of mutant opsin proteins associated with RP is reduced. 5 In methods of the invention, administering is preferably by topical administration (such as with an eye wash) or by systemic administration (including oral, intraocular injection or periocular injection). By way of preferred example, the ophthalmic condition to be treated is light toxicity, such as that resulting from ocular surgery, for example, retinal or cataract surgery. 10 Also encompassed is an ophthalmologic composition comprising an effective amount of compounds of the Formula I and/or Formula 11 in a pharmaceutically acceptable carrier, wherein said agent reversibly binds non covalently (for example, at or near the retinal binding pocket) to said opsin 15 protein to inhibit 11-cis-retinal binding in said pocket, preferably where the small molecule ligand is selective for opsin protein. The present invention further provides a screening method for identifying a small molecule ligand that reduces light toxicity in a mammalian 20 eye, comprising: (a) contacting a native opsin-protein with a test compound in the presence of 11-cis-retinal and under conditions that promote the binding of the test compound and the 11-cis-retinal to the native opsin protein; and (b) determining a reversible reduction in rate of formation of rhodopsin 25 relative to the rate when said test compound is not present, thereby identifying said test compound as a small molecule ligand that reduces light toxicity in a mammalian eye. In a preferred embodiment, said test compound is structurally related to a compound disclosed herein. 30 In a typical competition assay of the invention, a compound is sought that will tie up the retinal binding pocket of the opsin protein. Thus, the assay seeks to identify a small molecule opsin binding compound (one that will not be tightly regulated by the retina as to amount entering rod cells) that 63 competes with or prevents 1 1-cis-retinal or 9-cis-retinal from forming rhodopsin or isorhodopsin. Over time, this will slow the rate of formation of rhodopsin relative to the rate when 11-cis-retinal alone is present. In one embodiment, the assay is conducted in the presence of 11-cis-retinal, and the 5 rate of formation of rhodopsin is measured as a way of determining competition for the retinal binding pocket, for example, by determining the rate of increase in the 500 nm peak characteristic for rhodopsin. No antibodies for rhodopsin are required for this assay. A useful compound will exhibit a rate of rhodopsin formation that is at least about 2 to 5 fold lower than that observed 10 in the presence of 11 -cis-retinal when said test compound is not present. The compounds of the Formula I and/or Formula il may be administered along with other agents, including a mineral supplement, an anti inflammatory agent, such as a steroid, for example, a corticosteroid, and/or an 15 anti-oxidant. Among the corticosteroids useful for such administration are those selected from the group consisting of cortisone, hydrocortisone, prednisone, prednisolone, methylprednisolone, triamcinolone, betamethasone, beclamethasone and dexamethasone. Useful anti-oxidants include vitamin A, vitamin C and vitamin E. 20 The methods of the invention also contemplate reducing light toxicity by using at least one additional agent (in addition to the compounds of the Formula I and/or Formula I selected from the group consisting of a proteasomal inhibitor, an autophagy inhibitor, a lysosomal inhibitor, an 25 inhibitor of protein transport from the ER to the Golgi, an Hsp90 chaperone inhibitor, a heat shock response activator, a glycosidase inhibitor, and a histone deacetylase inhibitor, wherein the small molecule opsin binding and the additional compound are administered simultaneously or within fourteen days of each other in amounts sufficient to treat the subject. 30 In a particular example of the methods of the invention, the compounds of the Formula I and/or Formula 11 and the additional compound are administered within ten days of each other, within five days of each other, 64 within twenty-four hours of each other and preferably are administered simultaneously. In one example, the small molecule opsin binding and the additional compound are administered directly to the eye. Such administration may be intraocular or intravitrial. In other examples, the small molecule opsin 5 binding and the additional compound are each incorporated into a composition that provides for their long-term release, such as where the composition is part of a microsphere, nanosphere, nano emulsion or implant. As described herein, the compounds of the Formula I and/or Formula 11 10 useful in the methods of the invention are available for use alone or in combination with one or more additional compounds to treat or prevent conditions associated with excessive rhodopsin activation, such as light toxicity, for example, resulting from ocular surgical procedures. In one embodiment, compounds of the Formula I and/or Formula II of the invention is 15 administered without an additional active compound. In another embodiment, compounds of the Formula I and/or Formula II of the invention is used in combination and with another active compound (e.g., as discussed herein). In still another exemplary embodiment, compounds of the Formula I and/or Formula 11 are administered in combination with the proteasomal inhibitor 20 MG132, the autophagy inhibitor 3-methyladenine, a lysosomal inhibitor ammonium chloride, the ER-Golgi transport inhibitor brefeldin A, the Hsp9O chaperone inhibitor Geldamycin, the heat shock response activator Celastrol, the glycosidase inhibitor, and the histone deacetylase inhibitor Scriptaid, can be used to reduce formation of visual cycle products and cell apoptosis as a 25 result of excessive rhodopsin activation. As described herein, the compounds of the Formula I and/or Formula II useful in the methods of the invention are available for use alone or in combination with one or more additional compounds to treat or prevent the 30 aberrant processing and trafficking of mutant opsin proteins associated with rod cell death as a result of RP. In one embodiment, compounds of the Formula I and/or Formula II of the invention is administered without an additional active compound. In another embodiment, compounds of the 65 Formula I and/or Formula II of the invention is used in combination and with another active compound (e.g., as discussed herein). In still another exemplary embodiment, compounds of the Formula I and/or Formula 11 are administered in combination with the proteasomal inhibitor MG132, the 5 autophagy inhibitor 3-methyladenine, a lysosomal inhibitor ammonium chloride, the ER-Golgi transport inhibitor brefeldin A, the Hsp90 chaperone inhibitor Geldamycin, the heat shock response activator Celastrol, the glycosidase inhibitor, and the histone deacetylase inhibitor Scriptaid, can be used to reduce or prevent the rod cell death and resulting blindness 10 associated with RP. As described herein, the compounds of the Formula I and/or Formula i useful in the methods of the invention are available for use alone or in combination with one or more additional compounds to treat or prevent 15 conditions associated with production and accumulation of toxic visual cycle products derived from all-trans-retinal, such as lipofucin and A2E, for example, the blindness associated with wet or dry ARMD. In one embodiment, compounds of the Formula I and/or Formula II of the invention is administered without an additional active compound. In another embodiment, 20 compounds of the Formula I and/or Formula II of the invention is used in combination and with another active compound (e.g., as discussed herein). In still another exemplary embodiment, compounds of the Formula I and/or Formula Il are administered in combination with the proteasomal inhibitor MG132, the autophagy inhibitor 3-methyladenine, a lysosomal inhibitor 25 ammonium chloride, the ER-Golgi transport inhibitor brefeldin A, the Hsp90 chaperone inhibitor Geldamycin, the heat shock response activator Celastrol, the glycosidase inhibitor, and the histone deacetylase inhibitor Scriptaid, can be used to reduce formation of toxic visual cycle product metabolites and photo cell death as a result of dry ARMD. 30 In specific embodiments of the methods of the invention, the mis-folded opsin protein comprises a mutation in its amino acid sequence, for example, one of the mutations T17M, P347S or P23H, preferably P23H.

66 Preferably, in any of the methods of the invention, the opsin-binding agent binds to opsin in its retinal binding pocket. In one aspect, the present invention provides a method of inhibiting the 5 formation or accumulation of a visual cycle product, comprising contacting an opsin protein with a compound that reduces hydration of said opsin protein, preferably wherein said compound competes with one or more water molecules for binding to opsin. In specific embodiments of such methods, the compound binds chemically to the opsin protein, for example, through 10 hydrogen bonding. In specific examples of the methods of the invention, a compound useful therein may bind to opsin at any hydration site found within the retinal binding pocket of the opsin molecule so long as said binding excludes wholly, 15 or in part, the binding of one or more water molecules in said binding pocket. Preferably the compound used in such method binds so as to occupy the left side of the binding pocket as shown in Figure 1 and displace waters in hydration sites 5-20 (numbered circles in Figure 1), more preferably binds so that waters in hydration sites 5-20 are displaced, and waters at hydration sites 20 3 or 4 as shown in Figure 1 are displaced and replaced with functionality on the ligand that mimics the hydrogen bonding interactions that these waters are predicted to have with residues on the protein. A specific example of these methods contemplates binding of a 25 compound by chemical interaction with Cysl87 or Glul13 of the opsin protein. In separate embodiments thereof, said interaction is with Cys187 or said interaction is with Glu113 or is with both sites. A preferred mode of said interaction is hydrogen bonding. 30 In other specific examples, said interaction is with a carbonyl group on the opsin protein. In specific embodiments thereof, said carbonyl is on Cysl87 or Glu113 of said opsin protein. Separate embodiments include where the carbonyl is on Cys1 87 of the opsin protein or where the carbonyl is on Glul 13 67 of the opsin protein. In one embodiment, the carbonyl is in the gamma carboxyl group of GIu113 of the opsin protein. A preferred embodiment is where the interaction is through an amine, carboxamido or urea group on the compound. 5 While use of any of the compounds disclosed herein as a means of reducing hydration in the opsin binding pocket should be considered a preferred embodiment of such method, the reduction of formation of a visual cycle product by reducing the formation of rhodopsin is a general method of 10 the invention for reducing such visual cycle product formation, especially production of lipofuscin and/or A2E, and for treating an ophthalmic disease by reducing said hydration is a general aim of the invention and is not necessarily limited in scope only to the use of chemicals disclosed herein but may include use of other known or yet to be known chemical compounds so 15 long as they function in the methods of the invention and reduce hydration (i.e., binding of water) in the retinal binding pocket of opsin. It should be noted that the compounds disclosed herein for use in the methods of the invention may not function to reduce hydration in the retinal 20 binding pocket of opsin but may still function in one or more of the methods of the invention. For example, a compound of Formula I and/or Formula 11 may bind to an allosteric site on the protein thereby excluding retinal from the retinal binding site without necessarily decreasing hydration yet still reduce formation of a visual cycle product, such as lipofuscin and/or A2E, by virtue of 25 its excluding retinal from the binding pocket, thus non-covalently reducing the activity of the visual cycle. In embodiments of any of the compositions and methods of the invention, the opsin-binding agent (e.g., a non-retinoid binding agent) is 30 selective for binding to opsin. Such selectivity is not to be taken as requiring exclusivity that said agent may bind to other proteins as well as to opsin but its binding to opsin will be at least selective, whereby the binding constant (or dissociation constant) for binding to opsin will be lower than the average value 68 for binding to other proteins that also bind retinoids, such as retinal analogs. Preferably, opsin binding agents are non-retinoid opsin-binding agents that bind non-covalently to opsin. Preferably, the opsin binding agent binds at or near the opsin retinal binding pocket, where the native ligand, 11-cis-retinal, 5 normally binds. Without wishing to be bound by theory, in one embodiment the binding pocket accommodates retinal or an agent of the invention, but not both. Accordingly, when an agent of the invention is bound at or near the retinal binding pocket, other retinoids, such as 11-cis-retinal, are unable to bind to opsin. Binding of an agent of the invention inside the retinal binding 10 pocket of a mis-folded opsin molecule serves to direct formation of the native or wild-type conformation of the opsin molecule or to stabilize a correctly folded opsin protein, thereby facilitating insertion of the now correctly-folded opsin into the membrane of a rod cell. Again, without wishing to be bound by theory, said insertion may help to maintain the wild-type conformation of opsin 15 and the opsin-binding agent is free to diffuse out of the binding pocket, whereupon the pocket is available for binding to retinal to form light-sensitive rhodopsin. Other methods of the invention provide a means to restore 20 photoreceptor function in a mammalian eye containing a mis-folded opsin protein that causes reduced photoreceptor function, comprising contacting said mis-folded opsin protein with an opsin-binding agent (e.g., a non-retinoid) that reversibly binds (e.g., that binds non-covalently) at or near the retinal binding pocket. In other embodiments, binding of the opsin-binding agent to 25 the mis-folded opsin protein competes with 11 -cis-retinal for binding in said binding pocket. Desirably, binding of the opsin-binding agent restores the native conformation of said mis-folded opsin protein. In preferred embodiments, the mammalian eye is a human eye. In 30 additional embodiments, said contacting occurs by administering said opsin binding agent (e.g., non-retinoid) to a mammal afflicted with an ophthalmic condition, such as a condition characterized by reduced photoreceptor function. In various embodiments, the condition is the wet or dry form of 69 macular degeneration, diabetic RP, a retinal or macular dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity (e.g., due 5 to retinal surgery), or retinitis pigmentosa. The administration may be topical administration or by systemic administration, the latter including oral administration, intraocular injection or periocular injection. Topical administration can include, for example, eye drops containing an effective amount of an agent of the invention in a suitable pharmaceutical carrier. 10 In another embodiment, the present invention also provides a method of stabilizing a mutant opsin protein, comprising contacting said mutant opsin protein with a non-retinoid opsin-binding agent that reversibly binds non covalently (for example, at or in the retinal binding pocket) to said mutant 15 opsin protein to prevent retinoid binding in said binding pocket, thereby stabilizing said mutant opsin protein. The present invention also provides a method of ameliorating loss of photoreceptor function in a mammalian eye, comprising administering an 20 effective amount of an opsin-binding agent, such as a non-retinoid, to a mammal afflicted with a mutant opsin protein that has reduced affinity for 11 cis-retinal, whereby the opsin binding agent reversibly binds (e.g., non covalently) to the retinal binding pocket of said mutant opsin, thereby ameliorating loss of photoreceptor function in said mammalian eye. In one 25 embodiment, the contacting occurs by administering said opsin-binding agent to a mammal afflicted with said reduced photoreceptor function, wherein said administering may be by topical administration or by systemic administration, the latter including oral, intraocular injection or periocular injection, and the former including the use of eye drops containing an agent of the invention. 30 Such loss of photoreceptor function may be a partial loss or a complete loss, and where a partial loss it may be to any degree between 1% loss and 99% loss. In addition, such loss may be due to the presence of a mutation that causes mis-folding of the opsin, such as where the mutation is the P23H 70 mutation. In another embodiment, the opsin binding agent is administered to ameliorate an opthalmic condition related to the mislocalization of an opsin protein. In one embodiment, the invention provides for the treatment of a subject having the dry form of age-related macular degeneration, where at 5 least a portion of the opsin present in an ocular photoreceptor cell (e.g., a rod or cone cell) is mislocalized. The mislocalized protein fails to be inserted into the membrane of a photoreceptor cell, where its function is required for vision. Administration of the opsin binding agent to a subject having a mislocalized opsin protein rescues, at least in part, opsin localization. Accordingly, the 10 invention is useful to prevent or treat an ophthalmic condition related to opsin mislocalization or to ameliorate a symptom thereof. The present invention provides a method for treating and/or preventing an ophthalmic condition or a symptom thereof, including but not limited to, wet 15 or dry form of macular degeneration, retinitis pigmentosa, a retinal or macular dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity (e.g., due to retinal surgery), or retinitis pigmentosa in 20 a subject, such as a human patient, comprising administering to a subject afflicted with, or at risk of developing, one of the aforementioned conditions or another ophthalmic condition related to the expression of a misfolded or mislocalized opsin protein using a therapeutically effective amount of an opsin-binding agent, e.g., an agent that shows positive activity when tested in 25 any one or more of the screening assays of the invention. Such a method may also comprise administering to said subject at least one additional agent selected from the group consisting of a proteasomal inhibitor, an autophagy inhibitor, a lysosomal inhibitor, an 30 inhibitor of protein transport from the ER to the Golgi, an Hsp90 chaperone inhibitor, a heat shock response activator, a glycosidase inhibitor, and a histone deacetylase inhibitor, wherein the opsin-binding compound and the additional compound are administered simultaneously or within fourteen days 71 of each other in amounts sufficient to treat the subject. Here again the patient may comprise a mutation that affects protein folding where said mutation(s) causes mis-folding, e.g., in an opsin protein, 5 and may be any of the mutations recited elsewhere herein, such as a P23H mutation. In other embodiments, the patient has an ophthalmic condition that is related to the mislocalization of an opsin protein. The mislocalized opsin fails to insert into the membrane of a photoreceptor cell (e.g., a rod or cone cell). In general, this failure in localization would effect only a portion of the 10 opsin present in an ocular cell of a patient. In particular examples of the methods of the invention, the opsin binding compound and the additional compound are administered within ten days of each other, more preferably within five days of each other, even more 15 preferably within twenty-four hours of each other and most preferably are administered simultaneously. In one example, the opsin-binding compound and the additional compound are administered directly to the eye. Such administration may be intra-ocular. In other examples, the opsin-binding compound and the additional compound are each incorporated into a 20 composition that provides for their long-term release, such as where the composition is part of a microsphere, nanosphere, or nano emulsion. In one example, the composition is administered via a drug-delivery device that effects long-term release. Such methods also contemplate administering a vitamin A supplement along with an agent of the invention. 25 As described herein, the opsin-binding agents useful in the methods of the invention are available for use alone or in combination with one or more additional compounds to treat or prevent conditions associated with the wet or dry form of macular degeneration, retinitis pigmentosa, a retinal or macular 30 dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity (e.g., due to retinal surgery), retinitis pigmentosa or 72 another ophthalmic condition related to the expression of a misfolded or mislocalized opsin protein. In one embodiment, an opsin-hinding compound of the invention (e.g., a non-retinoid or a retinoid that fails to covalently bind to opsin) is administered to a subject identified as having or at risk of developing 5 such a condition. Optionally, the opsin binding agent is administered together with another therapeutic agent. In another embodiment, a non-retinoid opsin binding compound of the invention is used in combination with a synthetic retinoid (e.g., as disclosed in U.S. Patent Publication No. 2004-0242704), and optionally with another active compound (e.g., as discussed herein). In still 10 another exemplary embodiment, an opsin-binding compound is administered in combination with the proteasomal inhibitor MG132, the autophagy inhibitor 3-methyladenine, a lysosomal inhibitor, such as ammonium chloride, the ER Golgi transport inhibitor brefeldin A, the Hsp90 chaperone inhibitor Geldamycin, the heat shock response activator Celastrol, the glycosidase 15 inhibitor, and/or the histone deacetylase inhibitor Scriptaid, or any other agent that can stabilize a mutant P23H opsin protein in a biochemically functional conformation that allows it to associate with 1 1-cis-retinal to form rhodopsin. In specific embodiments, an opsin-binding compound is a non 20 polymeric (e.g., a small molecule, such as those disclosed herein for use in the methods of the invention) compound having a molecular weight less than about 1000 daltons, less than 800, less than 600, less than 500, less than 400, or less than about 300 daltons. In certain embodiments, a compound of the invention increases the amount (e.g., from or in a cell) of a stably-folded 25 and/or complexed mutant protein by at least 10%, 15%, 20%, 25%, 50%, 75%, or 100% compared to an untreated control cell or protein. Proteasomal inhibitors 30 The 26S proteasome is a multicatalytic protease that cleaves ubiquinated proteins into short peptides. MG-132 is one proteasomal inhibitor that may be used. MG- 132 is particularly useful for the treatment of light toxicity and other ocular diseases related to the accumulation of visual cycle 73 products (e.g., all-trans-retinal, A2E, lipofuscin), protein aggregation or protein misfolding. Other proteasomal inhibitors useful in combination with of the invention in the methods of the invention include lactocystin (LC), clasto lactocystin-beta-lactone, PSI (N-carbobenzoyl-Ile-Glu-(OtBu)-Ala-Leu-CHO), 5 MG-132 (N-carbobenzoyl-Leu-Leu-Leu-CHO), MG-115 (Ncarbobenzoyl-Leu Leu-Nva-CHO), MG-101 (N-Acetyl-Leu-Leu-norLeu-CHO), ALLM (NAcetyl Leu-Leu-Met-CHO), N-carbobenzoyl-Gly-Pro-Phe-leu-CHO, N-carbobenzoyl Gly-Pro-Ala-Phe-CHO, N-carbobenzoyl-Leu-Leu-Phe-CHO, and salts or analogs thereof. Other proteasomal inhibitors and their uses are described in 10 U.S. Patent No. 6,492,333. Autophagy inhibitors Autophagy is an evolutionarily conserved mechanism for the 15 degradation of cellular components in the cytoplasm, and serves as a cell survival mechanism in starving cells. During autophagy pieces of cytoplasm become encapsulated by cellular membranes, forming autophagic vacuoles that eventually fuse with lysosomes to have their contents degraded. Autophagy inhibitors may be used in combination with an opsin-binding or 20 opsin-stabilizing compound of the invention. Autophagy inhibitors useful in combination with a of the invention in the methods of the invention include, but are not limited to, 3-methyladenine, 3-methyl adenosine, adenosine, okadaic acid, N -mercaptopurine riboside (N 6 -MPR), an aminothiolated adenosine analog, 5-amino-4-imidazole carboxamide riboside (AICAR), 25 bafilomycin Al, and salts or analogs thereof. Lysosomal inhibitors The lysosome is a major site of cellular protein degradation. 30 Degradation of proteins entering the cell by receptor-mediated endocytosis or by pinocytosis, and of plasma membrane proteins takes place in lysosomes. Lysosomal inhibitors, such as ammonium chloride, leupeptin, trans epoxysaccinyl-L-leucylamide-(4-guanidino) butane, L-methionine methyl 74 ester, ammonium chloride, methylamine, chloroquine, and salts or analogs thereof, are useful in combination with an opsin-binding or opsin-stabilizing compound of the invention. 5 HSP90 chaperone inhibitors Heat shock protein 90 (Hsp9O) is responsible for chaperoning proteins involved in cell signaling, proliferation and survival, and is essential for the conformational stability and function of a number of proteins. HSP-90 10 inhibitors are useful in combination with an opsin-binding or opsin-stabilizing compound in the methods of the invention. HSP-90 inhibitors include benzoquinone ansamycin antibiotics, such as geldanamycin and 17 allylamino-17-demethoxygeldanamycin (17-AAG), which specifically bind to Hsp9O, alter its function, and promote the proteolytic degradation of substrate 15 proteins. Other HSP-90 inhibitors include, but are not limited to, radicicol, novobiocin, and any Hsp9O inhibitor that binds to the Hsp9O ATP/ADP pocket. Heat shock response activators 20 Celastrol, a quinone methide triterpene, activates the human heat shock response. In combination with an opsin-binding or opsin-stabilizing compound in methods of the invention, celastrol and other heat shock response activators are useful for the treatment of PCD. Heat shock response 25 activators include, but are not limited to, celastrol, celastrol methyl ester, dihydrocelastrol diacetate, celastrol butyl ester, dihydrocelastrol, and salts or analogs thereof. Histone deacetylase inhibitors 30 Regulation of gene expression is mediated by several mechanisms, including the post-translational modifications of histones by dynamic acetylation and deacetylation. The enzymes responsible for reversible 75 acetylationl/deacetylation processes are histone acetyltransferases (HATs) and histone deacetylases (HDACs), respectively. Histone deacetylase inhibitors include Scriptaid, APHA Compound 8, Apicidin, sodium butyrate, (-) Depudecin, Sirtinol, trichostatin A, and salts or analogs thereof. Such 5 inhibitors may be used in combination with compounds of the invention in the methods disclosed herein. Glycosidase inhibitors 10 Glycosidase inhibitors are one class of compounds that are useful in the methods of the invention, when administered in combination with an opsin-binding or opsin-stabilizing compound of the invention. Castanospermine, a polyhydroxy alkaloid isolated from plant sources, inhibits enzymatic glycoside hydrolysis. Castanospermine and its derivatives are 15 particularly useful for the treatment of light toxicity or of an ocular Protein Conformation Disorder, such as RP. Also useful in the methods of the invention are other glycosidase inhibitors, including australine hydrochloride, 6-Acetamido-6-deoxy-castanosperrnine, which is a powerful inhibitor of hexosaminidases, Deoxyfuconojirimycin hydrochloride (DFJ7), 20 Deoxynojirimycin (DNJ), which inhibits glucosidase I and II, Deoxygalactonojirimycin hydrochloride (DGJ), winch inhibits aX-D galactosidase, Deoxymannojirimycin hydrochloride (DM1), 2R,5R Bis(hydroxymethyl)-3R,4R-dihydroxypyrrolidine (DMDP), also known as 2,5 dideoxy-2,5-imino-D-mannitol, 1,4-Dideoxy-1,4-imino-D-mannitol 25 hydrochloride, (3R,4R,5R,6R)-3,4,5,6-Tetrahydroxyazepane Hydrochloride, which inhibits b-N-acetylglucosaminidase, 1,5-Dideoxy-1,5-imino-xylitol, which inhibits p-glucosidase, and Kifunensine, an inhibitor of mannosidase 1. Also useful in combination with an opsin-binding or opsin-stabilizing compound are N-butyldeoxynojirimycin (EDNJ), N-nonyl DNJ (NDND, N-hexyl DNJ (15TDNJ), 30 N-methyldeoxynojirimycin (MDNJ), and other glycosidase inhibitors known in the art. Glycosidase inhibitors are available commercially, for example, from Industrial Research Limited (Wellington, New Zealand) and methods of using them are described, for example, in U.S. Patent Nos. 4,894,388, 5,043,273, 76 5,103,008, 5,844,102, and 6,831,176; and in U.S. Patent Publication Nos. 20020006909. Pharmaceutical Compositions 5 The present invention features pharmaceutical preparations comprising compounds together with pharmaceutically acceptable carriers, where the compounds provide for the inhibition of visual cycle products, such as all trans-retinal or other products formed from 11-cis-retinal. Such preparations 10 have both therapeutic and prophylactic applications. In one embodiment, a pharmaceutical composition includes an opsin-binding or stabilizing compound (e.g., a compound identified using the methods of Example 1) or a pharmaceutically acceptable salt thereof; optionally in combination with at least one additional compound that is a proteasomal inhibitor, an autophagy 15 inhibitor, a lysosomal inhibitor, an inhibitor of protein transport from the ER to the Golgi, an Hsp90 chaperone inhibitor, a heat shock response activator, a glycosidase inhibitor, or a histone deacetylase inhibitor. The opsin-binding or opsin-stabilizing compound is preferably not a natural or synthetic retinoid. The opsin-binding or opsin-stabilizing compound and the additional compound 20 are formulated together or separately. Compounds of the invention may be administered as part of a pharmaceutical composition. The non-oral compositions should be sterile and contain a therapeutically effective amount of the opsin-binding or opsin-stabilizing compound in a unit of weight or volume suitable for administration to a subject. The compositions and 25 combinations of the invention can be part of a pharmaceutical pack, where each of the compounds is present in individual dosage amounts. The phrase "pharmaceutically acceptable" refers to those compounds of the present invention, compositions containing such compounds, and/or 30 dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

77 Non-oral pharmaceutical compositions of the invention to be used for prophylactic or therapeutic administration should be sterile. Sterility is readily accomplished by filtration through sterile filtration membranes (e.g., 0.2 pLm membranes), by gamma irradiation, or any other suitable means known to 5 those skilled in the art. Therapeutic opsin-binding or opsin-stabilizing compound compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle. These compositions ordinarily will be stored in unit or multi-dose containers, for example, sealed ampoules 10 or vials, as an aqueous solution or as a Iyophilized formulation for reconstitution. The compounds may be combined, optionally, with a pharmaceutically acceptable excipient. The components of the pharmaceutical compositions also are capable 15 of being co-mingled with the molecules of the present invention, and with each other, in a manner such that there is no interaction that would substantially impair the desired pharmaceutical efficacy. Compounds of the present invention can be contained in a 20 pharmaceutically acceptable excipient. The excipient preferably contains minor amounts of additives such as substances that enhance isotonicity and chemical stability. Such materials are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetate, lactate, tartrate, and other organic acids or their salts; tris 25 hydroxymethylaminomethane (TRIS), bicarbonate, carbonate, and other organic bases and their salts; antioxidants, such as ascorbic acid; low molecular weight (for example, less than about ten residues) polypeptides, e.g., polyarginine, polylysine, polyglutamate and polyaspartate; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as 30 polyvinylpyrrolidone (PVP), polypropylene glycols (PPGs), and polyethylene glycols (PEGs); amino acids, such as glycine, glutamic acid, aspartic acid, histidine, lysine, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, mannose, 78 sucrose, dextrins or sulfated carbohydrate derivatives, such as heparin, chondroitin sulfate or dextran sulfate; polyvalent metal ions, such as divalent metal ions including calcium ions, magnesium ions and manganese ions; chelating agents, such as ethylenediamine tetraacetic acid (EDTA); sugar 5 alcohols, such as mannitol or sorbitol; counterions, such as sodium or ammonium; and/or nonionic surfactants, such as polysorbates or poloxamers. Other additives may be included, such as stabilizers, anti-microbials, inert gases, fluid and nutrient replenishers (i.e., Ringer's dextrose), electrolyte replenishers, which can be present in conventional amounts. 10 The compositions, as described above, can be administered in effective amounts. The effective amount will depend upon the mode or administration, the particular condition being treated and the desired outcome. It may also depend upon the stage of the condition, the age and physical 15 condition of the subject, the nature of concurrent therapy, if any, and like factors well known to the medical practitioner. For therapeutic applications, it is that amount sufficient to achieve a medically desirable result. With respect to a subject suffering from, or at risk of developing, light 20 toxicity, such as that due to ocular surgery, an effective amount is an amount sufficient to reduce the rate or extent of formation and accumulation of visual cycle products, such as all-trans-retinal, or lipofuscin, or A2E as well as preventing photocell apoptosis as a result of excessive rhodopsin activation. Here, the compounds of the present invention would be from about 0.01 25 mg/kg per day to about 1000 mg/kg per day. It is expected that doses ranging from about 50 to about 2000 mg/kg will be suitable. Lower doses will result from certain forms of administration, such as intravenous administration. In the event that a response in a subject is insufficient at the initial doses applied, higher doses (or effectively higher doses by a different, more 30 localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of a composition of the present invention.

79 A variety of administration routes are available. The methods of the invention, generally speaking, may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of the active compounds without causing clinically 5 unacceptable adverse effects. In one preferred embodiment, a composition of the invention is administered intraocularly. Other modes of administration include oral, rectal, topical, intraocular, buccal, intravaginal, intracisternal, intracerebroventricular, intratracheal, nasal, transdermal, within/on implants, or parenteral routes. Compositions comprising a composition of the invention 10 can be added to a physiological fluid, such as to the intravitreal humor. For CNS administration, a variety of techniques are available for promoting transfer of the therapeutic across the blood brain barrier including disruption by surgery or injection, drugs which transiently open adhesion contact between the CNS vasculature endothelial cells, and compounds that facilitate 15 translocation through such cells. Oral administration can be preferred for prophylactic treatment because of the convenience to the patient as well as the dosing schedule. Pharmaceutical compositions of the invention can optionally further 20 contain one or more additional proteins as desired, including plasma proteins, proteases, and other biological material, so long as it does not cause adverse effects upon administration to a subject. Suitable proteins or biological material may be obtained from human or mammalian plasma by any of the purification methods known and available to those skilled in the art; from 25 supernatants, extracts, or lysates of recombinant tissue culture, viruses, yeast, bacteria, or the like that contain a gene that expresses a human or mammalian plasma protein which has been introduced according to standard recombinant DNA techniques; or from the fluids (e.g., blood, milk, lymph, urine or the like) or transgenic animals that contain a gene that expresses a 30 human plasma protein which has been introduced according to standard transgenic techniques. Pharmaceutical compositions of the invention can comprise one or 80 more pH buffering compounds to maintain the pH of the formulation at a predetermined level that reflects physiological pH, such as in the range of about 5.0 to about 8.0 (e.g., 6.0, 6.5, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.8). The pH buffering compound used in the aqueous liquid formulation can 5 be an amino acid or mixture of amino acids, such as histidine or a mixture of amino acids such as histidine and glycine. Alternatively, the pH buffering compound is preferably an agent which maintains the pH of the formulation at a predetermined level, such as in the range of about 5.0 to about 8.0, and which does not chelate calcium ions. Illustrative examples of such pH 10 buffering compounds include, but are not limited to, imidazole and acetate ions. The pH buffering compound may be present in any amount suitable to maintain the pH of the formulation at a predetermined level. Pharmaceutical compositions of the invention can also contain one or 15 more osmotic modulating agents, i.e., a compound that modulates the osmotic properties (e.g., tonicity, osmolality and/or osmotic pressure) of the formulation to a level that is acceptable to the blood stream and blood cells of recipient individuals. The osmotic modulating agent can be an agent that does not chelate calcium ions. The osmotic modulating agent can be any 20 compound known or available to those skilled in the art that modulates the osmotic properties of the formulation. One skilled in the art may empirically determine the suitability of a given osmotic modulating agent for use in the inventive formulation. Illustrative examples of suitable types of osmotic modulating agents include, but are not limited to: salts, such as sodium 25 chloride and sodium acetate; sugars, such as sucrose, dextrose, and mannitol; amino acids, such as glycine; and mixtures of one or more of these agents and/or types of agents. The osmotic modulating agent(s) maybe present in any concentration sufficient to modulate the osmotic properties of the formulation. 30 Compositions comprising an opsin-binding or opsin-stabilizing compound of the present invention can contain multivalent metal ions, such as calcium ions, magnesium ions and/or manganese ions. Any multivalent 81 metal ion that helps stabilize the composition and that will not adversely affect recipient individuals may be used. The skilled artisan, based on these two criteria, can determine suitable metal ions empirically and suitable sources of such metal ions are known, and include inorganic and organic salts. 5 Pharmaceutical compositions of the invention can also be a non aqueous liquid formulation. Any suitable non-aqueous liquid may be employed, provided that it provides stability to the active agents (a) contained therein. Preferably, the non-aqueous liquid is a hydrophilic liquid. Illustrative 10 examples of suitable non-aqueous liquids include: glycerol; dimethyl sulfoxide (DMSO); polydimethylsiloxane (PMS); ethylene glycols, such as ethylene glycol, diethylene glycol, triethylene glycol, polyethylene glycol ("PEG") 200, PEG 300, and PEG 400; and propylene glycols, such as dipropylene glycol, tripropylene glycol, polypropylene glycol ("PPG") 425, PPG 15 725, PPG 1000, PEG 2000, PEG 3000 and PEG 4000. Pharmaceutical compositions of the invention can also be a mixed aqueous/non-aqueous liquid formulation. Any suitable non-aqueous liquid formulation, such as those described above, can be employed along with any 20 aqueous liquid formulation, such as those described above, provided that the mixed aqueous/non-aqueous liquid formulation provides stability to the compound contained therein. Preferably, the non- aqueous liquid in such a formulation is a hydrophilic liquid. Illustrative examples of suitable non aqueous liquids include: glycerol; DMSO; EMS; ethylene glycols, such as 25 PEG 200, PEG 300, and PEG 400; and propylene glycols, such as PPG 425, PPG 725, PEG 1000, PEG 2000, PEG 3000 and PEG 4000. Suitable stable formulations can permit storage of the active agents in a frozen or an unfrozen liquid state. Stable liquid formulations can be stored at a temperature of at least -70 0 C, but can also be stored at higher temperatures 30 of at least 0 0 C, or between about 0 0 C and about 42 0 C, depending on the properties of the composition. It is generally known to the skilled artisan that proteins and polypeptides are sensitive to changes in pH, temperature, and a multiplicity of other factors that may affect therapeutic efficacy.

82 In certain embodiments a desirable route of administration can be by pulmonary aerosol. Techniques for preparing aerosol delivery systems containing polypeptides are well known to those of skill in the art. Generally, such systems should utilize components that will not significantly impair the 5 biological properties of the antibodies, such as the paratope binding capacity (see, for example, Sciarra and Cutie, "Aerosols," in Remington's Pharmaceutical Sciences 18th edition, 1990, pp 1694-1712; incorporated by reference). Those of skill in the art can readily modify the various parameters and conditions for producing polypeptide aerosols without resorting to undue 10 experimentation. Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of compositions of the invention, increasing convenience to 15 the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art. They include polymer base systems such as polylactides (U.S. Pat. No. 3,773,919; European Patent No. 58,481), poly(lactide-glycolide), copolyoxalates polycaprolactones, polyesteramides, polyorthoesters, poiyhydroxybutyric acids, such as poly-D-( 20 )-3-hydroxybutyric acid (European Patent No. 133,988), copolymers of L glutamic acid and gamma-ethyl-L-glutamate (Sidman, KR. et at, Biopolymers 22: 547-556), poly (2-hydroxyethyl methacrylate) or ethylene vinyl acetate (Langer, et al., J. Biomed. Mater. Res. 15:267-277; Langer, B.. Chem. Tech. 12:98-105), and polyanhydrides. 25 Other examples of sustained-release compositions include semi permeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Delivery systems also include non-polymer systems that are: lipids including sterols such as cholesterol, cholesterol esters and fatty acids 30 or neutral fats such as mono-, di- and tri-glycerides; hydrogel release systems such as biologically-derived bioresorbable hydrogel (i.e., chitin hydrogels or chitosan hydrogels); sylastic systems; peptide based systems; wax coatings; compressed tablets using conventional binders and excipients; partially filled 83 implants; and the like. Specific examples include, but are not limited to: (a) aerosional systems in which the agent is contained in a form within a matrix such as those described in 13.5. Patent Nos. 4,452,775, 4,667,014, 4,748,034 and 5,239,660 and (b) diffusional systems in which an active component 5 permeates at a controlled rate from a polymer such as described in U.S. Patent Nos. 3,832,253, and 3,854,480. Another type of delivery system that can be used with the methods and compositions of the invention is a colloidal dispersion system. Colloidal 10 dispersion systems include lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. Liposomes are artificial membrane vessels, which are useful as a delivery vector in vivo or in vitro. Large unilamellar vessels (LUV), which range in size from 0.2 - 4.0 ptm, can encapsulate large macromolecules within the aqueous interior and be 15 delivered to cells in a biologically active form (Fraley, R., and Papahadjopoulos, D., Trends Biochem. Sci. 6: 77-80). Liposomes can be targeted to a particular tissue by coupling the liposome to a specific ligand such as a monoclonal antibody, sugar, glycolipid, 20 or protein. Liposomes are commercially available from Gibco BRL, for example, as LIPOFECTIN TM and LIPOFECTACE T M , which are formed of cationic lipids such as N-[1-(2, 3 dioleyloxy)-propyl]-N,N,N trimethylammonium chloride (DOTMA) and dimethyl dioctadecylammonium bromide (DDAB). Methods for making liposomes are well known in the art and 25 have been described in many publications, for example, in DE 3,218,121; Epstein et al., Proc. Natl. Acad. Sci. (USA) 82:3688-3692 (1985); K. Hwang et al., Proc. Natl, Acad. Sci. (USA) 77:4030-4034 (1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641; Japanese Pat. Apple. 83-118008; U.S. Pat. Nos. 4,485,045 and 4,544,545; and EP 102,324. Liposomes also have 30 been reviewed by Gregoriadis, G., Trends Biotechnol., 3: 235-241. Another type of vehicle is a biocompatible microparticle or implant that is suitable for implantation into the mammalian recipient. Exemplary 84 bioerodible implants that are useful in accordance with this method are described in PCT International application no. PCTIUS/03307 (Publication No WO 95/24929, entitled "Polymeric Gene Delivery System"). PCT/US/0307 describes biocompatible, preferably biodegradable polymeric matrices for 5 containing an exogenous gene under the control of an appropriate promoter. The polymeric matrices can be used to achieve sustained release of the exogenous gene or gene product in the subject. The polymeric matrix preferably is in the form of a microparticle such 10 as a microsphere (wherein an agent is dispersed throughout a solid polymeric matrix) or a microcapsule (wherein an agent is stored in the core of a polymeric shell). Microcapsules of the foregoing polymers containing drugs are described in, for example, U.S. Patent 5,075,109. Other forms of the polymeric matrix for containing an agent include films, coatings, gels, 15 implants, and stents. The size and composition of the polymeric matrix device is selected to result in favorable release kinetics in the tissue into which the matrix is introduced. The size of the polymeric matrix further is selected according to the method of delivery that is to be used. Preferably, when an aerosol route is used the polymeric matrix and composition are encompassed 20 in a surfactant vehicle. The polymeric matrix composition can be selected to have both favorable degradation rates and also to be formed of a material, which is a bioadhesive, to further increase the effectiveness of transfer. The matrix composition also can be selected not to degrade, but rather to release by diffusion over an extended period of time. The delivery system can also be 25 a biocompatible microsphere that is suitable for local, site-specific delivery. Such microspheres are disclosed in Chickering, D.B., et al., Biotechnot. Bioeng, 52: 96-101; Mathiowitz, B., et at., Nature 386: 410-414. Both non-biodegradable and biodegradable polymeric matrices can be 30 used to deliver the compositions of the invention to the subject. Such polymers may be natural or synthetic polymers. The polymer is selected based on the period of time over which release is desired, generally in the order of a few hours to a year or longer. Typically, release over a period 85 ranging from between a few hours and three to twelve months is most desirable. The polymer optionally is in the form of a hydrogel that can absorb up to about 90% of its weight in water and further, optionally is cross-linked with multivalent ions or other polymers. 5 Exemplary synthetic polymers which can be used to form the biodegradable delivery system include: polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates, polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, polyvinyl 10 halides, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes and copolymers thereof, alkyl cellulose, hydroxyaikyl celluloses, cellulose ethers, cellulose esters, nitro celluoses, polymers of acrylic and methacrylic esters, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose 15 acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxylethyl cellulose, cellulose triacetate, cellulose sulfate sodium salt, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl mcthacrylate), poly(lauryl methacrylate), poly(phenyl 20 methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene, polypropylene, poly(ethylene glycol), poly(ethylene oxide), poly(ethylene terephthalate), poly(vinyl alcohols), poly(vinyl acetate), poly(vinyl chloride), polystyrene, poly(viny lpyrrolidone), and polymers of lactic acid and glycolic acid, polyanhydrides, 25 poly(ortho)esters, poly(butic acid), poly(valeric acid), and poly(lactide cocaprolactone), and natural polymers such as alginate and other polysaccharides including dextran and cellulose, collagen, chemical derivatives thereof (substitutions, additions of chemical groups, for example, alkyl, alkylene, hydroxylations, oxidations, and other modifications routinely 30 made by those skilled in the art), albumin and other hydrophilic proteins, zein and other prolamines and hydrophobic proteins, copolymers and mixtures thereof. In general, these materials degrade either by enzymatic hydrolysis or exposure to water in vivo, by surface or bulk erosion.

86 Methods of Ocular Delivery The compositions of the invention are particularly suitable for treating ocular diseases or conditions, such as light toxicity, in particular light toxicity 5 related to an ocular surgical procedure. In one approach, the compositions of the invention are administered through an ocular device suitable for direct implantation into the vitreous of the eye. The compositions of the invention may be provided in sustained 10 release compositions, such as those described in, for example, U.S. Pat. Nos. 5,672,659 and 5,595,760. Such devices are found to provide sustained controlled release of various compositions to treat the eye without risk of detrimental local and systemic side effects. An object of the present ocular method of delivery is to maximize the amount of drug contained in an 15 intraocular device or implant while minimizing its size in order to prolong the duration of the implant. See, e.g., U.S. Patents 5,378,475; 6,375,972, and 6,756,058 and U.S. Publications 20050096290 and 200501269448. Such implants may be biodegradable and/or biocompatible implants, or may be non-biodegradable implants. 20 Biodegradable ocular implants are described, for example, in U.S. Patent Publication No. 20050048099. The implants may be permeable or impermeable to the active agent, and may be inserted into a chamber of the eye, such as the anterior or posterior chambers or may be implanted in the 25 sclera, transchoroidal space, or an avascularized region exterior to the vitreous. Alternatively, a contact lens that acts as a depot for compositions of the invention may also be used for drug delivery. In a preferred embodiment, the implant may be positioned over an 30 avascular region, such as on the sclera, so as to allow for transcleral diffusion of the drug to the desired site of treatment, e.g. the intraocular space and macula of the eye. Furthermore, the site of transcleral diffusion is preferably in proximity to the macula. Examples of implants for delivery of a composition of 87 the invention include, but are not limited to, the devices described in U.S. Pat. Nos. 3,416,530; 3,828,777; 4,014,335; 4,300,557; 4,327,725; 4,853,224; 4,946,450; 4,997,652; 5,147,647; 164,188; 5,178,635; 5,300,114; 5,322,691; 5,403,901; 5,443,505; 5,466,466; 5,476,511; 5,516,522; 5,632,984; 5 5,679,666; 5,710,165; 5,725,493; 5,743,274; 5,766,242; 5,766,619; 5,770,592; 5,773,019; 5,824,072; 5,824,073; 5,830,173; 5,836,935; 5,869,079, 5,902,598; 5,904,144; 5,916,584; 6,001,386; 6,074,661; 6,110,485; 6,126,687; 6,146.366; 6,251,090; and 6,299,895, and in WO 01/30323 and WO 01/28474, all of which are incorporated herein by 10 reference. Examples include, but are not limited to the following: a sustained release drug delivery system comprising an inner reservoir comprising an effective amount of an agent effective in obtaining a desired local or systemic 15 physiological or pharmacological effect, an inner tube impermeable to the passage of the agent, the inner tube having first and second ends and covering at least a portion of the inner reservoir, the inner tube sized and formed of a material so that the inner tube is capable of supporting its own weight, an impermeable member positioned at the inner tube first end, the 20 impermeable member preventing passage of the agent out of the reservoir through the inner tube first end, and a permeable member positioned at the inner tube second end, the permeable member allowing diffusion of the agent out of the reservoir through the inner tube second end; a method for administering a compound of the invention to a segment of an eye, the 25 method comprising the step. of implanting a sustained release device to deliver the compound of the invention to the vitreous of the eye or an implantable, sustained release device for administering a compound of the invention to a segment of an eye; a sustained release drug delivery device comprising: a) a drug core comprising a therapeutically effective amount of at 30 least one first agent effective in obtaining a diagnostic effect or effective in obtaining a desired local or systemic physiological or pharmacological effect; b) at least one unitary cup essentially impermeable to the passage of the agent that surrounds and defines an internal compartment to accept the drug 88 core, the unitary cup comprising an open top end with at least one recessed groove around at least some portion of the open top end of the unitary cup; c) a permeable plug which is permeable to the passage of the agent, the permeable plug is positioned at the open top end of the unitary cup wherein 5 the groove interacts with the permeable plug holding it in position and closing the open top end, the permeable plug allowing passage of the agent out of the drug core, though the permeable plug, and out the open top end of the unitary cup; and d) at least one second agent effective in obtaining a diagnostic effect or effective in obtaining a desired local or systemic physiological or 10 pharmacological effect; or a sustained release drug delivery device comprising: an inner core comprising an effective amount of an agent having a desired solubility and a polymer coating layer, the polymer layer being permeable to the agent, wherein the polymer coating layer completely covers the inner core. 15 Other approaches for ocular delivery include the use of liposomes to target a compound of the present invention to the eye, and preferably to retinal pigment epithelial cells and/or Bruch's membrane. For example, the compound maybe complexed with liposomes in the manner described above, 20 and this compound/liposome complex injected into patients with an ophthalmic condition, such as light toxicity, using intravenous injection to direct the compound to the desired ocular tissue or cell. Directly injecting the liposome complex into the proximity of the retinal pigment epithelial cells or Bruch's membrane can also provide for targeting of the complex with some 25 forms of ocular PCD. In a specific embodiment, the compound is administered via intra-ocular sustained delivery (such as VITRASERT or ENVISION. In a specific embodiment, the compound is delivered by posterior subtenons injection. In another specific embodiment, microemulsion particles containing the compositions of the invention are delivered to ocular tissue to take up lipid 30 from Bruchs membrane, retinal pigment epithelial cells, or both. Nanoparticles are a colloidal carrier system that has been shown to improve the efficacy of the encapsulated drug by prolonging the serum half- 89 life. Polyalkylcyanoacrylates (PACAs) nanoparticles are a polymer colloidal drug delivery system that is in clinical development, as described by Stella et al, J. Pharm. Sci., 2000. 89: p. 1452-1464; Brigger et al., Tnt. J. Pharm., 2001. 214: p. 37-42; Calvo et al., Pharm. Res., 2001. 18: p. 1157-1166; and Li et al., 5 Biol. Pharm. Bull., 2001. 24: p. 662-665. Biodegradable poly (hydroxyl acids), such as the copolymers of poly (lactic acid) (PLA) and poly (lactic-co glycolide) (PLGA) are being extensively used in biomedical applications and have received FDA approval for certain clinical applications. In addition, PEG PLGA nanoparticles have many desirable carrier features including (i) that the 10 agent to be encapsulated comprises a reasonably high weight fraction (loading) of the total carrier system; (ii) that the amount of agent used in the first step of the encapsulation process is incorporated into the final carrier (entrapment efficiency) at a reasonably high level; (iii) that the carrier have the ability to be freeze-dried and reconstituted in solution without aggregation; (iv) 15 that the carrier be biodegradable; (v) that the carrier system be of small size; and (vi) that the carrier enhance the particles persistence. Nanoparticles are synthesized using virtually any biodegradable shell known in the art. In one embodiment, a polymer, such as poly (lactic-acid) 20 (PLA) or poly (lactic-co-glycolic acid) (PLGA) is used. Such polymers are biocompatible and biodegradable, and are subject to modifications that desirably increase the photochemical efficacy and circulation lifetime of the nanoparticle. In one embodiment, the polymer is modified with a terminal carboxylic acid group (COOH) that increases the negative charge of the 25 particle and thus limits the interaction with negatively charge nucleic acid aptamers. Nanoparticles are also modified with polyethylene glycol (PEG), which also increases the half-life and stability of the particles in circulation. Alternatively, the COOH group is converted to an N-hydroxysuccinimide (NHS) ester for covalent conjugation to amine-modified aptamers. 30 Biocompatible polymers useful in the composition and methods of the invention include, but are not limited to, polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene 90 terephthalates, polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, polyvinyl halides, poly(viny lpyrrolidone), polyglycolides, polysiloxanes, polyurethanes and copolymers thereof, alkyl cellulose, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, polymers of acrylic and methacrylic 5 esters, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxylethyl cellulose, cellulose triacetate, cellulose sulfate sodium salt poly-methyl methacrylate), poly(ethyl methacrylate), poly(butyl 10 methacrylate), poly(isobutyl methacrylate\ poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene, polypropylene poly(ethylene glycol), poly(ethylene oxide), poly(ethylene terephthalate), poly(vinyl 15 alcohols), polyvinyl acetate, polyvinyl chloride polystyrene, poly(vinyl pyrrolidone), polyhyaluronic acids, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylates), poly(ethyl methacrylates), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate) poly(isodecyl methaerylate), poly(lauryl 20 methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylatee), poly(isobutyl acrylate), poly(octadecyl acrylate) and combinations of any of these, In one embodiment, the nanoparticles of the invention include PEG-PLGA polymers. 25 Compositions of the invention may also be delivered topically. For topical delivery, the compositions are provided in any pharmaceutically acceptable excipient that is approved for ocular delivery. Preferably, the composition is delivered in drop form to the surface of the eye. For some application, the delivery of the composition relies on the diffusion of the 30 compounds through the cornea to the interior of the eye. Those of skill in the art will recognize that treatment regimens for using the compounds of the present invention to treat light toxicity or other 91 opthalmic conditions (e.g., RP) can be straightforwardly determined. This is not a question of experimentation, but rather one of optimization, which is routinely conducted in the medical arts. In vivo studies in nude mice often provide a starting point from which to begin to optimize the dosage and 5 delivery regimes. The frequency of injection will initially be once a week, as has been done in some mice studies. However, this frequency might be optimally adjusted from one day to every two weeks to monthly, depending upon the results obtained front the initial clinical trials and the needs of a particular patient. 10 Human dosage amounts can initially be determined by extrapolating from the amount of compound used in mice, as a skilled artisan recognizes it is routine in the art to modify the dosage for humans compared to animal models. For certain embodiments it is envisioned that the dosage may vary 15 from between about 1 mg compound/Kg body weight to about 5000 mg compound/Kg body weight; or from about 5 mg/Kg body weight to about 4000 mg/Kg body weight or from about 10mg/Kg body weight to about 3000 mg/Kg body weight; or from about 50mg/Kg body weight to about 2000 mg/Kg body weight; or from about 100 mg/Kg body weight to about 1000 mg/Kg body 20 weight; or from about 150 mg/Kg body weight to about 500 mg/Kg body weight. In other embodiments this dose maybe about 1, 5, 10, 25, 50,75, 100, 150, 10 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2500, 3000, 3500, 4000, 4500, 5000 25 mg/Kg body weight. in other embodiments, it is envisaged that lower does may be used, such doses may be in the range of about 5 mg compound/Kg body to about 20 mg compound/Kg body. In other embodiments the doses may be about 8, 10, 12, 14, 16 15 or 18 mg/Kg body weight. Of course, this dosage amount may be adjusted upward or downward, as is routinely done in 30 such treatment protocols, depending on the results of the initial clinical trials and the needs of a particular patient.

92 Screening Assays Useful compounds of the invention are compounds of the formual (1) that reversibly bind to a native or mutated opsin protein, such as in or near the 5 11-cis-retinal binding pocket. The non bleachable or slowly bleachable pigment rhodopsins formed from these small molecule opsin bindings will prevent light toxicity related to, for example, the accumulation of visual cycle products as well as apoptotic photocell death resulting from excessive rhodopsin stimulation. Such binding will commonly inhibit, if not prevent, 10 binding of retinoids, especially 11-cis-retinal, to the binding pocket and thereby reduce formation of visual cycle products, such as all-trans-retinal. Any number of methods are available for carrying out screening assays to identify such compounds. In one approach, an opsin protein is contacted with a candidate compound or test compound that is a non-retinoid in the presence 15 of 11-cis-retinal or retinoid analog and the rate or yield of formation of chromophore is determined. If desired, the binding of the non-retinoid to opsin is characterized. Preferably, the non-retinoid binding to opsin is non covalent and reversible. Thus, inhibition of rhodopsin formation by a non retinoid indicates identification of a successful test compound. An increase in 20 the amount of rhodopsin is assayed, for example, by measuring the protein's absorption at a characteristic wavelength (e.g., 498 nm for rhodopsin) or by measuring an increase in the biological activity of the protein using any standard method (e.g., enzymatic activity association with a ligand). Useful compounds inhibit binding of 11-cis-retinal (and formation of rhodopsin) by at 25 least about 10%, 15%, or 20%, or preferably by 25%, 50%, or 75%, or most preferably by up to 90% or even 100%. The efficacy of the identified compound is assayed in an animal model showing the effects of light toxicity. For example, the efficacy of compounds 30 disclosed herein have been demonstrated using transgenic mice that contain a mutant elaov 4 gene important in fatty acid synthesis and transgenic mice that produce a mutant ABCR protein that affects how all-trans-retinal is shuttled. The amount of lipofuscin produced in such mice was determined 93 using compounds of the invention and shown to be produced at a reduced rate resulting in slower accumulation of toxic visual cycle products. In either case, the cellular phenotype is the same and lipofuscin is accumulated at an accelerated rate when successful test compounds are not administered. 5 Alternatively, the efficacy of compounds useful in the methods of the invention may be determined by exposure of a mammalian eye to a high intensity light source prior to, during, or following administration of a test compound, followed by determination of the amount of visual cycle products 10 (e.g., all-trans retinal, A2E, or lipofuscin) formed as a result of exposure to the high intensity light source, wherein a compound of the invention will have reduced the amount of visual cycle products related to the exposure. In sum, preferred test compounds identified by the screening methods 15 of the invention are non-retinoids, are selective for opsin and bind in a reversible, non-covalent manner to opsin protein. In addition, their administration to transgenic animals otherwise producing increased lipofuscin results in a reduced rate of production or a reduced accumulation of lipofuscin in the eye of said animal. Compounds identified according to the methods of 20 the invention are useful for the treatment of light toxicity or other ophthalmic condition in a subject, such as a human patient. Combination Therapies 25 Compositions of the invention useful for the prevention of light toxicity, as well as AMD and retinitis pigmentosa, can optionally be combined with additional therapies as heretofore described. 30 94 EXAMPLES The following non-limiting examples further describe and enable one of ordinary skill in the art to make use of the invention. 5 Example 1: (E)-3-(2,6,6-Trimethylcyclohex-1-enyl)acrylic acid The title compound, obtained as a colorless crystalline solid (14.2 g, 52%), was prepared from p-ionone (26.7 g, 0.139 mol) according to the procedure of [Shimasaki, H.; Kagechika, H.; Fukasawa, H.; Kawachi, E.; 10 Shudo, K. Chem. Pharm. Bull. 1995, 43, 100-107]. Rf = 0.4 (25:75 ethyl acetate: hexanes); 'H-NMR (400 MHz, CDCl 3 ) 6 12.16 (br s, 1H), 7.56 (d, J = 16.0 Hz, 1H), 5.85 (d, J = 16.0 Hz, 1H), 2.08 (t, J = 6.0 Hz, 2H), 1.79 (s, 3H), 1.66-1.58 (m, 2H), 1.50-1.46 (m, 2H), 1.08 (s, 6H) ppm. 15 Example 2: (E)-3-(2,6,6-Trimethylcyclohex-1-enyl)acrylamide 2a. (E)-3-(2,6,6-Trimethylcyclohex-1-enyl)acryloyl chloride To a round bottom flask charged with (E)-3-(2,6,6-trimethylcyclohex-1 enyl) acrylic acid (1, 6.00 g, 3.00 mmol) in anhydrous dichloromethane (2.5 20 mL) under argon was added oxalyl chloride (0.50 mL, 5.50 mmol) dropwise via syringe. To this stirred solution was added two drops of N,N dimethylformamide (DMF) and the reaction was stirred at room temperature for 3 hours. The reaction mixture was concentrated in vacuo (40 0 C) to yield a yellow-brown oil which was carried forward without further purification. 25 2b. (E)-3-(2,6,6-Trimethylcyclohex-1 -enyl)acrylamide In a round bottom flask (E)-3-(2,6,6-trimethylcyclohex-1-enyl)acryloyI chloride (320 mg, 1.50 mmol) was dissolved in tetrahydrofuran (THF, 6.0 mL). The reaction mixture was cooled to 0*C and a solution of ammonium 30 hydroxide (0.4 mL) was added. The reaction mixture was warmed to room temperature while stirred for 4 hours. The crude reaction mixture was concentrated in vacuo (35 0 C) and purified by preparative plate thin layer chromatography (5:95 methanol: chloroform) to afford a yellow amorphous 95 solid (153 mg, 53%). Rf = 0.60 (3:97 methanol:chloroform); 1 H-NMR (400 MHz, CDC 3 ) 8 7.37 (d, J = 15.5 Hz, 1H), 6.42 (s, 1H), 5.83 (d, J = 15.5 Hz, 1H), 5.61 (s, 1H), 2.09-2.04 (m, 2H), 1.76 (s, 3H), 1.65-1.62 (m, 2H), 1.50 1.48 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 193.9 (MH*). 5 Example 3: (E)-N-Methyl-3-(2,6,6-trimethylcyclohex-1-enyl)acrylamide To a solution of (E)-3-(2,6,6-trimethylcyclohex-1-enyl) acrylic acid (1, 50.0 mg, 0.257 mmol) in DMF (1.0 mL) was added 2-(7-aza-1 H-benzotriazole 1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU, 97.7 mg, 0.257 10 mmol). The solution was stirred at room temperature for 30 minutes then diisopropylethylamine (66.4 mg, 0.514 mmol) and methylamine hydrochloride (17.4 mg, 0.257 mmol) was added to the reaction mixture. The reaction was then stirred at room temperature for 4 hours. The reaction was quenched with a 1M solution of hydrochloric acid (2 15 mL) and the biphasic mixture was separated. The organic layer was concentrated in vacuo (40*C) and the crude material loaded on to silica gel for purification via flash column chromatography running an isocratic eluent of 30% ethyl acetate in hexanes. The title compound was isolated as a white solid (56 mg, 86%). Mp = 84-88*C; Rf = 0.34 (50:50 ethyl acetate: hexanes); 20 1 H NMR (400 MHz, CDCl 3 ) 7.30 (d, J = 15.5, 1H), 5.74 (d, J = 15.5 Hz, 1H), 5.48 (d, J = 1.5 Hz, 1 H), 2.93 (d, J = 5.0 Hz, 3H), 2.05 (t, J = 6.0 Hz, 2H), 1.74 (s, 3H), 1.65-1.60 (m, 2H), 1.49 (dd, J = 7.5, 4.0 Hz, 2H), 1.06 (s, 6H); Mass spectrum (ESI +ve) m/z 208 (MH*). 25 Example 4: (E)-NN-Dimethyl-3-(2,6,6-trimethylcyclohex-1 enyl)acrylamide The title compound, obtained as an colorless oil (18 mg, 32%), was prepared from the product of Example 1 by following the procedure of Example 3 except dimethylamine (2.0 M solution in tetrahydrofuran) was 30 substituted for methylamine hydrochloride. Rf = 0.44 (50:50 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.34 (d, J = 15.6 Hz, 1H), 6.25 (d, J = 15.5 Hz, 1H), 3.10 (s, 3H), 3.05 (s, 3H), 2.05 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 96 1.64 (dd, J = 8.0, 4.0 Hz, 2H), 1.51-1.47 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 221 (MH*). Example 5: (E)-1-(Piperidin-1-yI)-3-(2,6,6-trimethylcyclohex-1-enyl)prop 5 2-en-1-one The title compound, obtained as an colorless oil (62 mg, 95%), was prepared from the product of Example 1 by following the procedure of Example 3 except piperidine was substituted for methylamine hydrochloride. Rf = 0.50 (40:60 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.30 10 (d, J = 13.5 Hz, 1H), 6.24 (d, J = 15.5 Hz, 1H), 3.70-3.45 (m, 4H), 2.04 (t, J = 6.0 Hz, 2H), 1.75 (s, 3H), 1.70-1.56 (m, 8H), 1.48 (dd, J = 7.5, 4.0 Hz, 2H), 1.06 (s, 6H); Mass spectrum (ESI +ve) m/z 262 (MH*). Example. 6: (E)-1-Morpholino-3-(2,6,6-trimethylcyclohex-1-enyl)prop-2 15 en-1-one The title compound, obtained as an colorless oil (60.0 mg, 89%), was prepared from the product of Example 1 by following the procedure of Example 3 except morpholine was substituted for methylamine hydrochloride and O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate 20 (HBTU) was substituted for 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate (HATU). Rf = 0.40 (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.36 (d, J = 15.5 Hz, 1H), 6.20 (d, J = 17.0 Hz, 1H), 3.77-3.47 (m, 7H), 3.10-3.02 (m, 1H), 2.03 (d, J = 5.5 Hz, 2H), 1.74 (s, 3H), 1.61 (dd, J = 7.5, 4.0 Hz, 2H), 1.51-1.42 (m, 2H), 25 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 264 (MH*). Example 7: (E)-1-(4-Methyl-1,4-diazepan-1-yl)-3-(2,6,6-trimethylcyclohex 1-enyl) prop-2-en-1 -one 30 7a. (E)-tert-Butyl 4.(3-(2,6,6-trimethylcyclohex-1 -enyl)acryloyl)-1,4 diazepane-1 -carboxylate 97 The title compound, obtained as an colorless oil (193 mg, 98%), was prepared from the product of Example 1 by following the procedure of Example 3 except tert-butyl 1,4-diazepane-1-carboxylate was substituted for methylamine hydrochloride and O-benzotriazole-NN,N', 5 tetramethyluronium hexafluorophosphate (HBTU) was substituted for 2-(7 aza-1H-benzotriazole-1-yi)-1,1,3,3-tetramethyluronium hexafluoro-phosphate (HATU). Rf = 0.40 (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.47-7.30 (m, 1H), 6.18 (d, J = 15.5 Hz, 1H), 3.73-3.31 (m, 8H), 2.04 (s, 2H), 1.95-1.83 (m, 2H), 1.75 (d, J = 12.0 Hz, 3H), 1.61 (dd, J = 12.0, 6.0 Hz, 2H), 10 1.45 (m, 11 H), 1.06 (s, 6H); Mass spectrum (ESI +ve) m/z 377 (MH*). 7b. (E)-1 -(1,4-Diazepan-1 -yI)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2 en-1-one To a solution of the product of Example 7a (160 mg, 0.425 mmol) in 15 dichloromethane (3 mL) was added dropwise a 2.0 M solution of hydrochloric acid in diethyl ether (0.43 mL, 0.85 mmol). The reaction mixture was stirred at room temperature for 18 hours. The title compound was isolated by flash column chromatography using a 10-25% methanol in dichloromethane solvent gradient to yield a dark yellow oil (115 mg, 87%). Rf = 0.30 (90:10 20 dichloromethane:methanol); 1 H NMR (400 MHz, CDCl 3 ) 8 7.78-7.23 (m, 2H), 6.40-6.01 (m, 2H), 4.32-4.13 (m, 1H), 3.81 (m, 4H), 3.22 (m, 4H), 2.12 (m, 3H), 1.84-1.25 (m, 9H), 1.08 (m, 5H), 0.92 (m, 3H); Mass spectrum (ESI +ve) m/z 277 (MH*). 25 7c. (E)-1-(4-Methyl-1,4-diazepan-1 -yI)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1 -one To a solution of (E)-1-(1,4-diazepan-1-yl)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1-one (7b, 50.0 mg, 0.181 mmol) and potassium carbonate (50.0 mg, 0.362 mmol) were dissolved in dichloromethane (3mL) at room 30 temperature. To this stirred solution was added iodomethane (25.7 mg, 0.181 mmol) dropwise via syringe. The reaction mixture was stirred for 18 hours at room temperature.

98 The reaction mixture was filtered to remove the excess potassium carbonate and then concentrated in vacuo (400C) to yield a crude solid. The crude material was purified using flash column chromatography (98:2 dichloromethane: methanol) to provide a pale yellow oil (16.0 mg, 31%). Rf = 5 0.75 (90:9:1 dichloromethane: methanol: ammonium hydroxide); 1 H NMR (400 MHz, CDCl 3 ) 8 7.34 (dd, J = 15.5, 7.5 Hz, 1H), 6.18 (dd, J = 15.5, 6.0 Hz, 1H), 3.80-3.57 (m, 4H), 2.74-2.50 (m, 4H), 2.38 (d, J = 8.5 Hz, 3H), 2.07-1.89 (m, 4H), 1.73 (s, 3H), 1.61 (td, J = 12.5, 6.0 Hz, 2H), 1.50-1.41 (m, 2H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 291 (MH*). 10 Example 8: (E)-1-(4-Ethyl-1,4-diazepan-1-yI)-3-(2,6,6-trimethylcyclohex-1 enyl) prop-2-en-1 -one The title compound, obtained as yellow oil (37 mg, 34%), was prepared from the product of Example 7b by following the procedure of Example 7c 15 except ethyliodide was substituted for iodomethane. Rf = 0.23 in (5:95 methanol: chloroform); 1 H NMR (400 MHz, CDCl 3 ) 6 7.34 (m, 1H), 6.19 (d, J = 15.5 Hz, 1H), 3.82-3.67 (m, 2H), 3.68-3.57 (m, 2H), 2.81-2.50 (m, 6H), 2.02 (m, 2H), 1.93 (m, 2H), 1.74 (s, 3H), 1.67-1.56 (m, 2H), 1.53-1.41 (m, 2H), 1.25 (s, 1H), 1.13-0.98 (m, 8H); Mass spectrum (ESI +ve) m/z 305 (MH*). 20 Example 9: (E)-1-(4-propyl-1,4-diazepan-1-yI)-3-(2,6,6-trimethylcyclohex 1-enyl) prop-2-en-1 -one 25 The title compound, obtained as yellow oil (12 mg, 21%), was prepared from the product of Example 7b by following the procedure of Example 7c except propyl bromide was substituted with iodomethane. Rf = 0.23 in (5:95 methanol: chloroform); 1 H NMR (400 MHz, CDCl 3 ) 6 7.32 (dd, J = 15.5, 7.5 Hz, 1H), 6.16 (m, 1H), 3.76-3.63 (m, 2H), 3.58 (t, J = 5.0 Hz, 2H), 2.78-2.66 30 (m, 2H), 2.67-2.56 (m, 3H), 2.51-2.36 (m, 2H), 2.00 (t, J = 6.0 Hz, 2H), 1.87 (m, 3H), 1.71 (s, 3H), 1.59 (m, 2H), 1.52-1.40 (m, 4H), 1.02 (s, 6H), 0.85 (m, 3H); Mass spectrum (ESI +ve) m/z 319 (MH*).

99 Example 10: (E)-1-(4-Acetyl-1,4-diazepan-1-yI)-3-(2,6,6-trimethylcyclohex 1-enyl) prop-2-en-1-one The title compound, obtained as a colorless oil (40.4 mg, 71%), was prepared from the product of Example 7b by following the procedure of 5 Example 7c except acetyl chloride was substituted for iodomethane. Rf = 0.80 (90:10 dichloromethane: methanol); 1 H NMR (400 MHz, CDCl 3 ) 6 7.47-7.27 (m, 1H), 6.15 (d, J = 15.5 Hz, 1H), 3.58 (m, 9H), 2.77 (s, 1H), 2.11-1.96 (m, 5H), 1.97-1.77 (m, 3H), 1.69 (d, J = 16.5 Hz, 3H), 1.58 (dd, J = 7.5, 4.0 Hz, 2H), 1.48-1.39 (m, 2H), 1.00 (s, 6H); Mass spectrum (ESI +ve) m/z 319 (MH*). 10 Example 11: (E)-1 -(4-Propionyl-1,4-diazepan-1 -yl)-3-(2,6,6 trimethylcyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as a colorless oil (56.1 mg, 93%), was prepared from the product of Example 7b by following the procedure of 15 Example 7c except propionyl chloride was substituted for iodomethane. Rf = 0.25 (75:25 dichloromethane: ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) 8 7.37 (q, J = 15.5 Hz, 1H), 6.17 (dd, J = 15.5, 6.0 Hz, 1H), 3.76-3.44 (m, 8H), 2.38-2.27 (m, 2H), 2.02 (s, 2H), 1.94-1.82 (m, 2H), 1.73 (s, 3H), 1.59 (d, J = 6.0 Hz, 2H), 1.50-1.41 (m, 2H), 1.14 (td, J = 14.5, 7.5 Hz, 3H), 1.03 (s, 6H); 20 Mass spectrum (ESI +ve) m/z 333 (MH*). Example 12: (E)-1-(4-(2,2,2-Trifluoroacetyl)-1,4-diazepan-1-yl)-3-(2,6,6 trimethyl cyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as a colorless oil (40.5 mg, 60%), was 25 prepared from the product of Example 7b by following the procedure of Example 7c except trifluoroacetic anhydride was substituted for iodomethane. Rf = 0.75 (75:25 dichloro- methane: ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) 8 7.41 (m, 1H), 6.17 (dd, J = 15.5, 6.5 Hz, 1H), 3.64 (m, 8H), 2.09-1.91 (m, 4H), 1.74 (s, 3H), 1.68-1.56 (m, 2H), 1.48 (dd, J = 7.5, 4.0 Hz, 2H), 1.05 (s, 30 6H); Mass spectrum (ESI +ve) m/z 373 (MH*). Example 13: (E)-4-(3-(2,6,6-Trimethylcyclohex-1 -enyl)acryloyl)-1,4 diazepane-1 -carboxamide 100 The title compound, obtained as yellow oil (20 mg, 21%), was prepared from the product of Example 7b by following the procedure of Example 7c except trimethylsilyl isocyanate was substituted for iodomethane. Rf = 0.57 (100% ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) 8 7.39 (m, 1H), 6.18 (d, J = 5 15.0 Hz, 1H), 5.19-4.66 (m, 2H), 3.84-3.32 (m, 8H), 2.02 (m, 2H), 1.73 (s, 3H), 1.60 (m, 2H), 1.46 (m, 2H), 1.25 (m, 2H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 320 (MH*). Example 14: (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 10 1,4-diazepane-1 -carboxamide (E)-1 -(1,4-Diazepan-1 -yl)-3-(2,6,6-trimethylcyclohex-1 -enyl)prop-2-en 1-one (7b, 50.0 mg, 0.181 mmol) and potassium carbonate (100 mg, 0.362 mmol) were dissolved in anhydrous dichloromethane (5 mL) under argon at room temperature. To this stirred solution was added 4-nitrophenyl 15 chloroformate (73.0 mg,- 0.362 mmol). The reaction mixture was stirred for 18 hours at room temperature. The reaction mixture was transferred to a microwave vial and the solvent was removed in vacuo. To the residue was added methylamine (56.2 mg, 1.81 mmol) and the sealed microwave vial was heated at 1000C for 30 20 minutes in a microwave reactor. The contents of the reaction vial were then poured into 10 mL of water, extracted with dichloromethane (3 x 10 mL) and the combined organic layers were dried over sodium sulfate. The solvent was removed in vacuo (400C) to yield a white solid. The crude material was purified using flash column chromatography (98:2 dichloromethane: methanol) 25 to provide a colorless oil (31 mg, 0.092 mmol, 25%). Rf = 0.23 (98:2 dichloromethane: methanol); 1 H NMR (400 MHz, CDCi 3 ) 8 7.37 (dd, J = 15.5, 15.5 Hz, 1H), 6.18 (dd, J = 15.5, 6.0 Hz, 1H), 4.48-4.38 (m, 1H), 3.76-3.25 (m, 8H), 2.81 (s, 3H), 2.08-1.82 (m, 4H), 1.74 (s, 3H), 1.68-1.60 (m, 4H), 1.48 1.42 (m, 2H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 334 (MH*). 30 101 Example 15: (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 1,4-diazepane-1 -carboxamide The title compound, obtained as colorless oil (63 mg, 100%), was prepared from the product of Example 7b by following the procedure of 5 Example 7c except ethyl isocyanate was substituted for iodomethane. Rf = 0.1 (80:20 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 5 7.40 (t, J = 15.5 Hz, 1H), 6.21 (dd, J = 15.5, 6.5 Hz, 1H), 4.38 (m, 1H), 3.71 (d, 2H), 3.54 (m, 4H), 3.34-3.24 (m, 2H), 2.83 (s, 2H), 2.06 (t, J = 6.0 Hz, 2H), 2.03-1.96 (m, 1H), 1.90 (t, J = 6.0 Hz, 1H), 1.7 (s, 3H), 1.64 (m, 2H), 1.52-1.46 (m, 2H), 1.16 10 (t, J = 7.0 Hz, 3H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 348 (MH*). Example 16: (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 1,4-diazepane-1 -carboxamide 15 The title compound, obtained as a pale yellow oil (61.0 mg, 93%), was prepared from the product of Example 7b by following the procedure of Example 7c except propyl isocyanate was substituted for iodomethane. Rf = 0.15 (50:50 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCla) 5 7.37 (t, J = 15.5 Hz, 1H), 6.18 (dd, J = 15.5, 7.0 Hz, 1H), 4.51-4.33 (m, 2H), 3.74 (t, J = 20 5.5 Hz, 1H), 3.62 (t, J = 5.5 Hz, 2H), 3.57-3.44 (m, 4H), 3.34 (t, J = 6.0 Hz, 1H), 3.15 (m, 3H), 2.08-1.93 (m, 3H), 1.86 (dd, J = 12.0, 6.0 Hz, 1H), 1.74 (s, 4H), 1.67-1.42 (m, 7H), 1.04 (s, 6H), 0.91 (t, J = 7.5 Hz, 5H); Mass spectrum (ESI +ve) m/z 350.3 (MH*). 25 Example 17: (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)-1,4-diazepane-1 -carboxamide The title compound, obtained as a pale yellow oil (66.0 mg, 99%), was prepared from the product of Example 7b by following the procedure of Example 7c except isopropyl isocyanate was substituted for iodomethane. Rf 30 = 0.70 (90:10 dichloro-methane: methanol); 1 H NMR (400 MHz, CDC 3 ) 8 7.36 (t, J = 16.0 Hz, 1H), 6.18 (dd, J = 15.5, 6.5 Hz, 1H), 4.29 (m, 1H), 3.95 (dd, J = 12.5, 6.0 Hz, 1H), 3.58 (m, 7H), 3.33 (s, 1H), 2.79 (d, J = 2.5 Hz, 1H), 2.11- 102 1.78 (m, 4H), 1.73 (s, 3H), 1.61 (dd, J = 5.5, 3.5 Hz, 2H), 1.52-1.38 (m, 3H), 1.12 (m, 6H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 362 (MH*). Example 18: (E)-Methyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl)-1,4 5 diazepane-1 -carboxylate The title compound, obtained as a colorless solid (40.1 mg, 67%), was prepared from the product of Example 7b by following the procedure of Example 7c except methyl chloroformate was substituted for iodomethane. Rf = 0.45 (75:25 dichloromethane: ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) 10 8 7.36 (t, J = 17.0 Hz, 1H), 6.15 (t, J = 12.5 Hz, 1H), 3.75-3.34 (m, 11H), 2.02 (t, J = 6.0 Hz, 2H), 1.86 (m, 3H), 1.73 (s, 3H), 1.60 (m, 2H), 1.51-1.38 (m, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 335 (MH*). Example 19: (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 15 1,4-diazepane-1 -carboxylate The title compound, obtained as pale yellow oil (61 mg, 98%), was prepared from the product of Example 7b by following the procedure of Example 7c except ethyl chloroformate was substituted for iodomethane. Rf = 0.1 (25:75 ethyl acetate: dichloromethane); 1 H NMR (400 MHz, CDC 3 ) 5 7.34 20 (d, 1H), 6.16 (dd, J = 15.5, 6.5 Hz, 1H), 4.52 (s, 1H), 3.75-3.41 (m, 7H), 3.37 3.15 (m, 3H), 2.05-1.78 (m, 4H), 1.71 (s, 3H), 1.63-1.53 (m, 2H), 1.48-1.39 (m, 2H), 1.10 (t, J = 7.0 Hz, 3H), 1.01 (s, 6H); Mass spectrum (ESI +ve) m/z 349 (MH*). 25 Example 20: (E)-1-(4-(Methylsulfonyl)-1,4-diazepan-1-yl)-3-(2,6,6 trimethylcyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as colorless oil (48 mg, 75%), was prepared from the product of Example 7b by following the procedure of Example 7c except methanesulfonyl chloride was substituted for 30 iodomethane. Rf = 0.7 (10:90 methanol: dichloromethane); 1 H NMR (400 MHz, CDCl 3 ) 8 7.41 (m, 1H), 6.25-6.11 (m, 1H), 3.75 (m, 4H), 3.41 (m, 4H), 2.84 (s, 3H), 1.98 (m, 2H), 1.74 (s, 3H), 1.65-1.54 (m, 2H), 1.51-1.39 (m, 4H), 1.09-0.98 (s, 6H); Mass spectrum (ESI +ve) m/z 355 (MH*).

103 Example 21: (E)-1-(4-(Ethylsulfonyl)-1,4-diazepan-1-yl)-3-(2,6,6 trimethylcyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as pale yellow oil (58 mg, 86%), was 5 prepared from the product of Example 7b by following the procedure of Example 7c except ethanesulfonyl chloride was substituted for iodomethane. Rf = 0.75 (10:90 methanol: dichloromethane); 1 H NMR (400 MHz, CDCl 3 ) 6 7.40 (m, 1H), 6.18 (m, 1H), 3.72 (m, 5H), 3.53-3.30 (m, 2H), 2.99 (m, 2H), 2.78 (s, 2H), 2.08-1.89 (m, 4H), 1.73 (s, 3H), 1.45 (m, 2H), 1.31 (t, J = 7.5 Hz, 10 2H), 1.21 (t, J = 7.5 Hz, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 369 (MH*). Example 22: (E)-1-(4-(Trifluoromethylsulfonyl)-1,4-diazepan-1-yl)-3-( 2

,

6

,

6 tri-methylcyclohex-1 -enyl)prop-2-en-1 -one 15 The title compound, obtained as colorless oil (22 mg, 30%), was prepared from the product of Example 7b by following the procedure of Example 7c except trifluoromethanesulfonyl chloride was substituted for iodomethane. Rf = 0.25 (25:75 ethyl acetate: dichloromethane); 1 H NMR (400 MHz, CDCI 3 ) 8 7.45 (m, 1H), 6.17 (m, 1H), 3.67 (m, 8H), 2.09-1.95 (m, 4H), 20 1.74 (s, 3H), 1.61 (m, 2H), 1.53-1.44 (m, 2H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 409 (MH*). Example 23: (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 1,4-diazepane-1 -carbothioamide 25 The title compound, obtained as pale yellow oil (50 mg, 79%), was prepared from the product of Example 7b by following the procedure of Example 7c except methyl thioisocyanate was substituted for iodomethane. Rf = 0.15 (25:75 ethyl acetate: dichloromethane); 1 H NMR (400 MHz, CDCl 3 ) 6 7.35 (t, J = 16.0.Hz, 1H), 6.19 (m, 1H), 5.87 (m, 1H), 4.17 (t, J = 5.0 Hz, 1H), 30 3.98-3.72 (m, 5H), 3.57 (m, 3H), 3.12 (d, J = 4.0 Hz, 3H), 2.78 (s, 2H), 1.98 (m, 4H), 1.73 (s, 3H), 1.64-1.55 (m, 2H), 1.46 (m, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 350 (MH*).

104 Example 24: (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 1,4-diazepane-1 -carbothioamide The title compound, obtained as pale yellow oil (66 mg, 100%), was prepared from the product of Example 7b by following the procedure of 5 Example 7c except ethyl thioisocyanate was substituted for iodomethane. Rf = 0.28 (80:20 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 8 7.43 (t, J = 15.5 Hz, 1H), 6.23 (m, 1H), 5.48 (s, 1H), 4.20 (t, J = 5.0 Hz, 1H), 4.02-3.90 (m, 2H), 3.82-3.76 (m, 1H), 3.71 (m, 2H), 3.60 (m, 3H), 2.83 (s, 2H), 2.07 (m, 2H), 2.03-1.94 (m, 1H), 1.77 (s, 3H), 1.69-1.58 (m, 4H), 1.54-1.47 (m, 2H), 10 1.26 (t, J = 7.0 Hz, 3H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 364 (MH*). Example 25: (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 1,4-diazepane-1 -carbothioamide 15 The title compound, obtained as pale yellow oil (64 mg, 94%), was prepared from the product of Example 7b by following the procedure of Example 7c except propyl thioisocyanate was substituted for iodomethane. Rf = 0.55 (25:75 ethyl acetate: dichloromethane); 1 H NMR (400 MHz, CDCl 3 ) 8 7.38 (t, J = 16.5 Hz, 1H), 6.21 (m, 1H), 5.67 (m, 1H), 4.18 (m, 1H), 3.99-3.74 20 (m, 4H), 3.65-3.50 (m, 5H), 2.09-1.93 (m, 4H), 1.75 (s, 3H), 1.68-1.56 (m, 4H), 1.48 (m, 2H), 1.05 (s, 6H), 0.95 (t, J = 7.5 Hz, 3H); Mass spectrum (ESI +ve) m/z 378 (MH*). Example 26: (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 25 enyl)acryloyl)-1,4-diazepane-1 -carbothioamide The title compound, obtained as pale yellow oil (64 mg, 94%), was prepared from the product of Example 7b by following the procedure of Example 7c except isopropyl thioisocyanate was substituted for iodomethane. Rf = 0.5 (25:75 ethyl acetate: dichloromethane); 1 H NMR (400 MHz, CDC 3 ) 6 30 7.38 (t, J = 16.5 Hz, 1H), 6.21 (m, 1H), 5.34 (m, 7.5 Hz, 1H), 4.64 (m, 1H), 4.13 (m, 1H), 3.83 (m, 4H), 3.57 (m, 3H), 2.08-1.92 (m, 4H), 1.75 (s, 3H), 1.62 (m, 2H), 1.51-1.43 (m, 2H), 1.24 (d, J = 6.5 Hz, 6H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 378 (MH*).

105 Example 27: (E)-1-(4-Methylpiperazin-1-yI)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1-one The title compound, obtained as a yellow oil (112 mg, 81%) was prepared from the product of Example 1 by following the procedure of 5 Example 6 except N-methylpiperazine was substituted for morpholine. Rf = 0.28 (90:10 dichloromethane: methanol); 1 H NMR (400 MHz, CD 2

CI

2 ) 6 7.26 (d, 1H, J = 16 Hz), 6.25 (d, 1H, J = 16 Hz), 3.74-3.47 (m, 4H), 2.41-2.38 (m, 4H), 2.30 (s, 3H), 2.08 (t, 2H, J = 6.4 Hz), 1.77 (s, 3H), 1.69-1.63 (m, 2H), 1.53-1.50 (m, 2H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 277.2 (MH*). 10 Example 28: (E)-1-(4-Ethylpiperazin-1-yI)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1-one 28a. (E)-tert-Butyl 4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) 15 piperazine-1 -carboxylate The title compound, obtained as a yellow oil (3.60 g, 99%), was prepared from the product of Example 1 by following the procedure of Example 3 except tert-butyl piperazine-1-carboxylate was substituted for methylamine hydrochloride and O-benzotriazole-N,N,N', 20 tetramethyluronium hexafluorophosphate (HBTU) was substituted for 2-(7 aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluoro-phosphate (HATU). Rf = 0.21 in (20:80 ethyl acetate:hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.38 (d, J = 15.5 Hz, 1H), 6.21 (d, J = 13.0 Hz, 1H), 3.70-3.48 (m, 8H), 2.06 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.67-1.62 (m, 2H), 1.49 (s, 11H), 25 1.10 (s, 6H); Mass spectrum (ESI +ve) m/z 363 (MH*). 28b. (E)-1-(Piperazin-1-yl)-3-(2,6,6-trimethylcyclohex-1-enyl)prop-2-en 1-one The title compound, obtained as a yellow oil (250 mg, 96%) was 30 prepared from the product of Example 28a by following the procedure of example 7b. Rf = 0.36 (91:8:1 dichloromethane: methanol: ammonium hydroxide); 1 H NMR (400 MHz, CD 2

CI

2 ) 6 7.25 (d, J = 16.0 Hz, 1H), 6.24 (d, J = 16.0 Hz, 1H), 3.70-3.45 (m, 4H), 2.88-2.83 (m, 4H), 2.07 (t, J = 6.0 Hz, 2H), 106 1.77 (s, 3H), 1.69-1.63 (m, 2H), 1.53-1.50 (m, 2H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 263 (MH*). 28c. (E)-1-(4-Ethylpiperazin-1-yI)-3-(2,6,6-trimethylcyclohex-1 5 enyl)prop-2-en-1-one The title compound, obtained as a yellow oil (21 mg, 38%) was prepared from the product of Example 28b following the procedure of Example 7c except ethyl bromide was substituted for iodomethane, acetonitrile was substituted for dichloromethane and the reaction was heated 10 at 50 *C instead of room temperature. R f = 0.28 (95:5 dichloromethane: methanol); 1 H NMR (400 MHz, CDCla) 6 7.30 (d, J = 16.0 Hz, 1H), 6.23 (d, J = 16.0 Hz, 1H), 3.80-3.60 (m, 4H), 2.55-2.42 (m, 6H), 2.07 (t, J = 6.5 Hz, 2H), 1.78 (s, 3H), 1.69-1.62 (m, 2H), 1.53-1.50 (m, 2H), 1.12 (t, J = 6.0 Hz, 3H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 291 (MH*). 15 Example 29: (E)-1 -(4-Propylpiperazin-1 -yl)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1-one The title compound, obtained as a clear oil (3.5 mg, 6%) was prepared from the product of Example 28b following the procedure of Example 7c 20 except propyl iodide was substituted for iodomethane. Rf = 0.53 (90:10 dichloromethane: methanol); 1 H NMR (400 MHz, CDCla) 8 7.35 (d, J = 16.0 Hz, 1H), 6.22 (d, J = 16.0 Hz, 1H), 3.79-3.46 (m, 4H), 2.95-2.84 (m, 4H), 2.05 (t, J = 6.0 Hz, 2H), 1.76 (s, 3H), 1.71 (s, 2H), 1.63 (m, 2H), 1.53-1.45 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 305 (MH*). 25 Example 30: (E)-1-(4-Acetylpiperazin-1-yl)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1-one The title compound, obtained as a clear oil (52 mg, 90%) was prepared from the product of Example 28b following the procedure of Example 7c 30 except that acetyl chloride was substituted for iodomethane. Rf = 0.16 (98:2 dichloromethane: methanol); 1 H NMR (400 MHz, CD 2

CI

2 ) 6 7.32 (d, J = 16.0 Hz, 1H), 6.25 (d, J = 16.0 Hz, 1H), 3.79-3.46 (m, 8H), 2.14-2.03 (m, 5H), 1.78 107 (s, 3H), 1.70-1.61 (m, 2H), 1.55-1.50 (m, 2H), 1.09 (s, 6H); Mass spectrum (ESI +ve) m/z 305 (MH*). Example 31: (E)-1 -(4-Propionylpiperazin-1 -yl)-3-(2,6,6-trimethylcyclohex 5 1-enyl) prop-2-en-1 -one The title compound, obtained as a clear oil (31 mg, 51%) was prepared from the product of Example 28b following the procedure of Example 7c except that propionyl chloride was substituted for iodomethane. Rf = 0.10 (98:2 dichloromethane: methanol); 'H NMR (400 MHz, CDC 3 ) 6 7.40 (d, J = 10 16.0 Hz, 1H), 6.22 (d, J = 16.0 Hz, 1H), 3.81-3.47 (m, 8H), 2.40 (dt, J = 7.5, 5.0 Hz, 4H), 2.07 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.64 (m, 2H), 1.53-1.46 (m, 2H), 1.23-1.14 (m, 2H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 319 (MH*). Example 32: (E)-1 -(4-(2,2,2-Trifluoroacetyl) piperazin-1 -yI)-3-( 2

,

6

,

6 15 trimethylcyclo-hex-1 -enyl)prop-2-en-1 -one The title compound, obtained as a clear oil (65 mg, 96%) was prepared from the product of Example 28b following the procedure of Example 7c except that trifluoroacetic anhydride was substituted for iodomethane. Rf = 0.25 (98:2 dichloro-methane: methanol); 1 H NMR (400 MHz, CDCl 3 ) 6 7.42 (d, 20 J = 16.0 Hz, 1H), 6.20 (d, J = 16.0 Hz, 1H), 3.70 (m, 8H), 2.10-2.01 (m, 2H), 1.77 (s, 3H), 1.64 (m 2H), 1.53-1.45 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 359 (MH*). Example 33: (E)-4-(3-(2,6,6-Trimethylcyclohex-1-enyl)acryloyl)piperazine 25 1-carboxamide The product of Example 28b (50 mg, 0.189 mmol) and triethylamine (79 ptL, 0.567 mmol) were dissolved in anhydrous dichloromethane (4 mL) at room temperature under argon and stirred for 5 minutes. To this stirred reaction mixture was added trimethylsilyl isocyanate (76 ptL, 0.567 mmol). The 30 reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was poured into a saturated aqueous solution of ammonium chloride (10 mL) and was extracted with dichloromethane (3X10 mL). The combined organic phases were dried over sodium sulfate and the 108 concentrated in vacuo. The product was purified by flash column chromatography to yield the title compound as a white solid (60 mg, quantitative). Mp = 144 *C; Rf = 0.31 (96:4 dichloromethane: methanol); 1 H NMR (300 MHz, CDCla) 8 7.39 (d, J = 16.0 Hz, 1H), 6.21 (d, J = 16.0 Hz, 1H), 5 4.61-4.60 (m, 2H), 3.90-3.40 (m, 8H), 2.06 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.66-1.62 (m, 2H), 1.51-1.47 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 306 (MH*). Example 34: (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 10 enyl)acryloyl)piperazine-1 - carboxamide The title compound, obtained as a clear oil (51 mg, 85%) was prepared from the product of Example 28b following the procedure of Example 14. Rf = 0.50 (50:50 ethyl acetate: hexane); 1 H NMR (400 MHz, CDC3l) 8 7.42 (d, 1H, J = 16.0 Hz), 6.20 (d, 1H, J = 16.0 Hz), 4.45 (br s, 1H), 3.80-3.30 (m, 8H), 2.88 15 (s, 3H), 2.09 (t, 2H, J = 6.4 Hz)), 1.77 (s, 3H), 1.70-1.64 (m 2H), 1.53-1.45 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 320 (MH*). Example 35: (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)piperazine-1- carboxamide 20 The title compound, obtained as a white solid (25 mg, 40%) was prepared from the product of Example 28b following the procedure of Example 7c except that ethyl isocyanate was substituted for iodomethane. Mp = 125-128 *C; Rf = 0.50 (2:1 ethyl acetate: dichloromethane); 1 H NMR (400 MHz, CDCl 3 ) 6 7.41 (d, J = 16.0 Hz, 1H), 6.22 (d, J = 16.0 Hz, 1H), 4.40 (br s, 25 1H), 3.82-3.30 (m, 10H), 2.13 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.70-1.64 (m 2H), 1.53-1.45 (m, 2H), 1.18 (t, J = 6.0 Hz, 3H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 334 (MH*). Example 36: (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1 30 enyl)acryloyl)piperazine-1- carboxamide The title compound, obtained as a clear oil (42 mg, 57%) was prepared from the product of Example 28b following the procedure of Example 7c except that n-propyl isocyanate was substituted for iodomethane. Rf = 0.21 109 (98:2 dichloromethane: methanol); 1 H NMR (400 MHz, CDCl 3 ) S 7.39 (d, J = 16.0 Hz, 1H), 6.21 (d, J = 16.0 Hz, 1H), 4.48 (s, 1H), 3.84-3.33 (m, 8H), 3.24 (q, J = 6.0 Hz, 2H), 2.06 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.68-1.46 (m, 9H), 1.07 (s, 6H), 0.95 (t, J = 7.5 Hz, 3H); Mass spectrum (ESI +ve) m/z 348 5 (MH*). Example 37: (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)piperazine-1- carboxamide The title compound, obtained as a white solid (62 mg, 85%) was 10 prepared from the product of Example 28b following the procedure of Example 7c except that isopropyl isocyanate was substituted for iodomethane. Mp = 158-159 *C; Rf = 0.32 (98:2 dichloro-methane: methanol); 1 H NMR (400 MHz, CDCl 3 ) a 7.37 (d, J = 16.0 Hz, 1H), 6.20 (de J = 16.0 Hz, 1H), 4.30 (d, J = 7.0 Hz, 1H), 3.99 (m, 1H), 3.82-3.31 (m, 9H), 2.05 (t, J = 6.0 15 Hz, 2H), 1.76 (s, 3H), 1.67-1.59 (m, 2H), 1.49 (m, 2H), 1.17 (d, J = 6.5 Hz, 6H), 1.06 (s, 6H); Mass spectrum (ESI +ve) m/z 348 (MH*). Example 38: (E)-Methyl 4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) piperazine-1 -carboxylate 20 The title compound, obtained as a colorless oil (34.0 mg, 55%), was prepared from the product of example 28b by following the procedure of Example 7c except methyl chloroformate was substituted for iodomethane. Rf = 0.25 in (20:80 ethyl acetate: dichloromethane); 1 H-NMR (400 MHz, CDCl 3 ) a 7.31 (d, J = 15.5 Hz, 1H), 6.23 (d, J = 15.5 Hz, 1H), 3.80-3.48 (m, 8H), 2.12 25 2.01 (m, 2H), 1.76 (s, 3H), 1.73-1.56 (m, 5H), 1.54-1.50 (m, 2H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 321 (MH*). Example 39: (E)-Ethyl 4-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) piperazine-1 -carboxylate 30 The title compound, obtained as a colorless oil (58.0 mg, 91%), was prepared from the product of example 28b by following the procedure of Example 7c except ethyl chloroformate was substituted for iodomethane. Rf = 0.12 in (20:80 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) a 7.48 (d, J 110 = 15.5 Hz, 1H), 6.21 (d, J = 15.5 Hz, 1H), 4.11-4.38 (m, 2H), 3.78-3.51 (m, 8H), 2.08 (m, 2H), 2.07 (s, 3H), 1.72-1.61 (m, 2H), 1.56-1.49 (m, 2H), 1.32 (m, 3H) 1.09 (s, 6H); Mass spectrum (ESI +ve) m/z 335 (MH*). 5 Example 40: (E)-1 -(4-(Methylsulfonyl)piperazin-1 -yl)-3-( 2

,

6

,

6 trimethylcyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as a white solid (38.2 mg, 59%), was prepared from the product of example 28b by following the procedure of Example 7c except methylsulfonyl chloride was substituted for iodomethane. 10 Mp = 121-123*C; Rf= 0.39 in (1:99 methanol: hexanes); 1 H-NMR (400 MHz,

CD

2

CI

2 ) 8 7.35 (d, J = 12.0 Hz, 1H), 6.25 (d, J = 12.0 Hz, 1H), 3.75 (br m,4H), 3.25 (m, 4H), 2.78 (s, 3H), 2.08-2.01 (m, 2H), 1.85 (s, 3H), 1.65-1.60 (m, 2H), 1.50-1.45 (m, 2H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 341 (MH*). 15 Example 41: (E)-1-(4-(Ethylsulfonyl)piperazin-1-yl)-3-( 2

,

6

,

6 trimethylcyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as a white solid (35.0 mg, 52%), was prepared from the product of example 28b by following the procedure of Example 7c except ethylsulfonyl chloride was substituted for iodomethane. 20 Mp = 125-127*C; Rf = 0.15 in (2:98 methanol:hexanes); 'H-NMR (400 MHz, CDCl 3 ) 8 7.42 (d, J = 12.0 Hz, 1H), 6.22 (d, J = 12.0 Hz, 1H), 3.85-3.70 (m, 4H), 3.40-3.30 (m, 4H), 2.99 (q, J = 6.0 Hz, 2H), 2.08-2.01 (m, 2H), 1.70-1.65 (m 2H), 1.60 (s, 3H), 1.55-1.50 (m, 2H), 1.40 (t, J = 6.0 Hz, 3H), 1.06 (s, 6H); Mass spectrum (ESI +ve) m/z 355 (MH*). 25 Example 42: (E)-1 -(4-(Trifluoromethylsulfonyl)piperazin-1 -yl)-3-(2,6,6 trimethyl-cyclohex-1 -enyl)prop-2-en-1 -one The title compound, obtained as a colorless oil (2.50 mg, 2.8%), was prepared from the product of example 28b by following the procedure of 30 Example 7c except trifluoromethylsulfonyl chloride was substituted for iodomethane. Rf = 0.75 in (20:80 ethyl acetate: dichloromethane); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.45 (d, J = 15.5 Hz, 1H), 6.22 (d, J = 15.5 Hz, 1H), 3.99- 111 3.42 (m, 8H), 2.15 (s, 3H), 1.74-1.58 (m, 2H), 1.56-1.48 (m, 2H), 1.25-1.31 (m, 2H),1.09 (s, 6H); Mass spectrum (ESI +ve) m/z 395 (MH*). Example 43: (E)-N-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 5 enyl)acryloyl)piperazine-1-carbothioamide The title compound, obtained as a white solid (38.2 mg, 61%), was prepared from the product of example 28b by following the procedure of Example 7c except methyl isothiocyanate was substituted for iodomethane. Mp = 55-57*C; Rf = 0.37 in (65:35 ethyl acetate: dichloromethane); 1 H-NMR 10 (400 MHz, CDCl 3 ) 5 7.44 (d, J = 15.5 Hz, 1H), 6.22 (d, J = 15.5 Hz, 1H), 5.98 (s, 1H), 4.18 (s, 2H), 3.89-3.68 (m, 6H), 3.19 (s, 3H), 2.12-2.08 (m, 2H), 1.75 (s, 3H), 1.72-1.61 (m, 2H) 1.52-1.49 (m, 2H), 1.10 (s, 6H); Mass spectrum (ESI +ve) m/z 336 (MH*). 15 Example 44: (E)-N-Ethyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)piperazine-1 -carbothioamide The title compound, obtained as a white solid (42.0 mg, 63%), was prepared from the product of example 28b by following the procedure of Example 7c except ethyl isothiocyanate was substituted for iodomethane. Mp 20 = 133-135*C; Rf = 0.33 in (50:50 ethyl acetate: dichloromethane); 1 H-NMR (400 MHz, CDC 3 ) 5 7.42 (d, J = 12.0 Hz, 1H), 6.20 (d, J = 12.0 Hz, 1H), 5.50 (br s, 1H), 4.25-4.10 (m, 2H), 3.95-3.65 (m 8H), 2.12-2.08 (m, 2H), 1.75 (s, 3H), 1.68-1.58 (m, 2H) 1.55-1.49 (m, 2H), 1.30 (t, J = 6.0 Hz, 1H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 350 (MH*). 25 Example 45: (E)-N-Propyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)piperazine-1 -carbothioamide The title compound, obtained as a white solid (37.9 mg, 55%), was prepared from the product of example 28b by following the procedure of 30 Example 7c except propyl isothiocyanate was substituted for iodomethane. Mp = 128-130*C; Rf = 0.53 in (50:50 ethyl acetate: dichloromethane); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.46 (d, J = 15.5 Hz, 1H), 6.23 (d, J = 15.5 Hz, 1H), 5.59 112 (s, 1H), 4.19 (s, 2H), 3.91-3.64 (m, 8H), 2.09 (s, 2H), 1.86-1.62 (m, 7H), 1.49 (s, 2H), 1.13-0.93 (m, 9H); Mass spectrum (ESI +ve) m/z 364 (MH*). Example 46: (E)-N-Isopropyl-4-(3-(2,6,6-trimethylcyclohex-1 5 enyl)acryloyl)piperazine-1 -carbothioamide The title compound, obtained as a white solid (35.2 mg, 51%), was prepared from the product of example 28b by following the procedure of Example 7c except isopropyl isothiocyanate was substituted for iodomethane. Mp = 147-148'C; Rf = 0.51 in (50:50 ethyl acetate: dichloromethane); 'H-NMR 10 (400 MHz, CDCl 3 ) 8 7.48 (d, J = 15.5 Hz, 1H), 6.22 (d, J = 15.5 Hz, 1H), 5.25 (s, 1H), 4.77-4.62 (m, 1H), 4.18 (s, 1H), 3.92-3.63 (m, 6H), 2.18-2.06 (m, 2H), 1.78 (s, 3H), 1.71-1.63 (m, 3H) 1.48-1.50 (m, 2H), 1.29 (m, 6H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 364 (MH*). 15 Example 47: (S,E)-1 -(3-Hydroxypyrrolidin-1 -yl)-3-( 2

,

6

,

6 trimethylcyclohex-1-enyl) prop-2-en-1 -one The title compound, obtained as a clear oil (196 mg, 97%) was prepared from the product of Example 1 following the procedure of Example 3 except (S)-pyrrolidin-3-ol was substituted for methylamine hydrochloride and 20 acetonitrile was substituted for dichloromethane. Rf = 0.20 (ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) & 7.36 (m, 1H), 6.08 (dd, J = 15.5, 8.5, 1H), 4.54 (m, 1H), 3.76-3.58 (m, 4H), 3.03-2.88 (m, 1H), 2.05 (m, 4H), 1.76 (s, 3H), 1.63 (m, 2H), 1.49 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 264 (MH*). 25 Example 48: (S,E)-1-(3-(2,6,6-Trimethylcyclohex-1 enyl)acryloyl)pyrrolidin-3-yl carbamate Trichloroacetyl isocyanate (63 piL, 0.53 mmol) was added to a solution of the product of Example 47 (70 mg, 0.26 mmol). The solution was stirred overnight at room temperature and then quenched by treatment with water 30 (0.5 mL). Ethyl acetate (20 mL) was added to dilute the reaction mixture, and the organic phase was extracted with water (20 mL). The organic layer was separated and dried over magnesium sulfate and the solvent was removed in vacuo. The desired product was isolated by preparative plate thin layer 113 chromatography (100% EtOAc) to yield a clear oil (23 mg, 15%). Rf = 0.42 (ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) 8 7.40.(d, J = 15.0, 1H), 6.80 (dd, J = 15.5, 1H), 5.30 (m, 1H), 4.74 (m, 2H), 3.72-3.66 (m, 4H), 3.62 (m, 2H), 2.17 (m, 2H), 2.06 (s, 3H), 1.77 (d, J = 5.5, 2H), 1.63 (m, 2H), 1.08 (s, 6H); Mass 5 spectrum (ESI +ve) m/z 307 (MH*). Example 49: (E)-1 -(3-Aminopyrrolidin-1 -yI)-3-(2,6,6-trimethylcyclohex-1 enyl)prop-2-en-1 -one hydrochloride 10 49a. (E)-tert-Butyl 1-(3-(2,6,6-trimethylcyclohex-1-enyl)acryloyl) pyrrolidin-3-yi carbamate The title compound, obtained as white solid (210 mg, 75%), was prepared from the product of Example 1 following the procedure of Example 3 except tert-butyl pyrrolid i n-3-ylcarbamate was substituted for methylamine 15 hydrochloride. The crude product was carries forward without further purification. Rf = 0.2 in (40:60 ethyl acetate: hexanes); Mass spectrum (ESI +ve) m/z 363 (MH*). 49b. (E)-1-(3-Aminopyrrolidin-1-yI)-3-(2,6,6-trimethylcyclohex-1 20 enyl)prop-2-en-1-one The product of Example 49a was dissolved in a 4N solution of hydrochloric acid in 1,4-dioxane (2 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated in vacuo to approximately 0.5 mL. The crude product was added via pipette to diethyl ether (50 mL) where a 25 white precipitate was formed. The precipitate was filtered, washed with ether and dried under reduced pressure to yield the desired product as a white solid (70 mg, 84%). Mp = 188 0 C; 1 H NMR (400 MHz, DMSO-d 6 ) 8 8.40-8.35 (m, 3H), 7.17 (m, 1H), 6.15 (m, 1H), 3.79 (m, 2H), 3.62 (m, 2H), 2.26 (m, 1H), 2.04 (m, 4H), 1.73 (s, 3H), 1.58 (m, 2H), 1.45 (m, 2H), 1.03 (s, 6H); Mass 30 spectrum (ESI +ve) m/z 263 (MH*).

114 Example 50: (E)-1-(1-(3-(2,6,6-Trimethylcyclohex-1-enyl)acryloyl) pyrrolidin-3-yl) urea The title compound, obtained as colorless oil (6.7 mg, 9%), was prepared from the product of Example 49b by following the procedure of 5 Example 48. Rf = 0.25 (90:10 chloroform: methanol); 1 H NMR (400 MHz, CD1 3 ) 8 7.35 (d, J = 15.5, 1 H), 6.50 (m, 1 H), 6.08 (t, J = 15.5, 1 H), 5.04 (br s, 2H), 4.32 (m, 1H), 3.61 (m, 4H), 2.06-2.05 (m, 4H), 1.76 (s, 3H), 1.62 (m, 2H), 1.48 (m, 2H), 1.07 (m, 6H); Mass spectrum (ESI +ve) m/z 306 (MH*). 10 Example 51: 4-(3-(2,6,6-Trimethylcyclohex-1-enyl)propanoyl)piperazine 1-carboxamide 51a. 1 -(Piperazin-1 -yI)-3-(2,6,6-trimethylcyclohex-1 -enyl)propan-1 -one Example 28b (100 mg, 0.381 mmol) was dissolved in anhydrous 15 methanol (10 mL) under argon and stirred at room temperature. To this stirred reaction mixture was added magnesium turnings (83.0 mg, 3.41 mmol). The reaction mixture was stirred under argon at room temperature for 48 hours. Methanol was removed in vacuo and the residue was taken up in water 20 (20 mL). The aqueous phase was extracted with chloroform (3 x 10 mL) and the combined organic phases were dried over sodium sulfate. The solvent was removed in vacuo to provide a yellow oil (62 mg crude). The product was purified by preparative thin layer chromatography to yield the title compound, as clear oil (4.2 mg, 4%). Rf = 0.39 (95:5 dichloromethane: methanol); 1 H 25 NMR (400 MHz, CDCla) 8 3.66-3.60 (m, 2H), 3.50-3.43 (m, 2H), 2.88 (d, J = 4.5 Hz, 4H), 2.36 (s, 3H), 1.93 (t, J = 6.0 Hz, 2H), 1.64-1.55 (m, 5H), 1.44 (dd, J = 7.5, 4.0 Hz, 2H), 1.29 (m, 1H), 1.17-1.10 (m, 1H), 1.02 (s, 6H); Mass spectrum (ESI +ve) m/z 265 (MH*). 30 51b. 4-(3-(2,6,6-Trimethylcyclohex-1-enyl)propanoyl)piperazine-1 carboxamide The title compound, obtained as a white solid (3.0 mg, 61%) was prepared from the product of Example 51a following the procedure of 115 Example 33. Rf = 0.45 (95:5 dichloromethane:methanol); 1 H NMR (400 MHz, CDCl 3 ) 6 4.53 (s, 2H), 3.74-3.66 (m, 2H), 3.52 (s, 4H), 3.40 (s, 2H), 2.38 (s, 3H), 1.93 (t, J = 6.0 Hz, 2H), 1.67-1.55 (m, 6H), 1.48-1.41 (m, 2H), 1.33-1.26 (m, 2H), 1.14 (m, 2H), 1.02 (s, 6H); Mass spectrum (ESI +ve) m/z 308 (MH*). 5 Example 52: (S,E)-2-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)piper-azine-1 -carboxamide 52a. (S)-tert-Butyl 4-ca rbamoyl-3-methyl pi perazine-1 -carboxylate 10 The title compound, obtained as a yellow foam (480 mg, quant.) was prepared from (S)-1-Boc-3-methylpiperazine following the procedure of Example 33. [aMD = 14 0 (c = 0.005, EtOH); Rf = 0.52 (95:5 dichloromethane: methanol +0.1% (v/v) ammonium hydroxide); 1 H NMR (400 MHz, CDCl 3 ) 6 4.81 (s, 2H), 3.97 (m, 5H), 3.09 (m, 3H), 2.99-2.73 (m, 1H), 1.47 (s, 9H), 1.42 15 1.36 (m, 1H), 1.18 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 244 (MH*). 52b. (S)-2-Methylpiperazine-1-carboxamide The title compound, obtained as a yellow oil (480 mg) was prepared from the product of Example 52a following the procedure of Example 7b. The 20 crude product was carried forward without purification. Rf = 0.1 (95:5 dichloromethane: methanol +0.1% (v/v) ammonium hydroxide); Mass spectrum (ESI +ve) m/z 144 (MH*). 52c. (S,E)-2-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 25 enyl)acryloyl)piperazine-1-carboxamide The title compound, obtained as a clear oil (315 mg, 56%) was prepared from the product of Example 1 using the procedure of Example 3 except the product of Example 52b was substituted for methylamine hydrochloride. [aD = 16 0 (c = 0.005, CHCIA); Rf = 0.43 (95:5 dichloromethane: 30 methanol +0.1% (v/v) ammonium hydroxide); 1 H NMR (400 MHz, CDCl 3 ) 6 7.41 (d, J = 15.0 Hz, 1H), 6.29-6.10 (m, 1H), 4.57 (s, 3H), 4.41-4.23 (m, 1H), 4.12-3.55 (m, 3H), 3.51 (d, J = 5.0 Hz, 1H), 3.22 (s, 3H), 2.94-2.79 (m, 1H), 116 2.06 (t, J = 6.0 Hz, 2H), 1.76 (s, 3H), 1.71-1.59 (m, 3H), 1.50 (m, 2H), 1.21 (s, 3H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 320 (MH*). Example 53: (RE)-2-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 5 enyl)acryloyl)piper-azine-1-carboxamide 53a. (R)-tert-Butyl 4-carbamoyl-3-methylpiperazine-1 -carboxylate The title compound, obtained as a yellow foam (377 mg, 65%) was prepared from (R)-1-Boc-3-methylpiperazine following the procedure of 10 Example 7b. [aX]D = -35 0 (c = 0.005, EtOH); Rf = 0.48 (95:5 dichloromethane: methanol +0.1% (v/v) ammonium hydroxide); 1 H NMR (400 MHz, CDCl 3 ) 8 4.50 (s, 2H), 4.23-3.52 (m, 5H), 3.10 (m, 2H), 3.01-2.78 (m, 1H), 1.63 (s, 1H), 1.49 (s, 10H), 1.21 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 244 (MH*). 15 53b. (R)-2-Methylpiperazine-1-carboxamide The title compound, obtained as a yellow oil (552 mg) was prepared from the product of Example 53a following the procedure of example 7b. The crude product was carried forward without purification. Rf = 0.1 (95:5 20 dichloromethane: methanol +0.1% (v/v) ammonium hydroxide); Mass spectrum (ESI +ve) m/z 144 (MH*). 53c. (RE)-2-Methyl-4-(3-(2,6,6-trimethylcyclohex-1 enyl)acryloyl)piperazine-1-carboxamide 25 The title compound, obtained as a colorless amorphous solid (120 mg, 31%) was prepared from the product of Example 1 using the procedure of Example 3 except the product of Example 53b was substituted for methylamine hydrochloride. [aX]D = -11 0 (c = 0.005, CHC1 3 ); Rf = 0.33 (93:7 dichloromethane: methanol +0.1% (v/v) ammonium hydroxide); 1 H NMR (400 30 MHz, CDCl 3 ) 6 7.38 (d, J = 15.5 Hz, 1H), 6.29-6.07 (m, 1H), 4.55 (s, 2H), 4.38-4.21 (m, 1H), 3.94-3.09 (m, 6H), 2.04 (t, J = 6.0 Hz, 2H), 1.74 (s, 3H), 1.66-1.57 (m, 2H), 1.50-1.43 (m, 5H), 1.40 (d, J = 6.0 Hz, 2H), 1.18 (s, 3H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 320 (MH*).

117 Example 54: (E)-N 2 -Methyl-4-(3-(2,6,6-trimethylcyclohex-1 -en-1 yl)acryloyl)piperazine-1,2-dicarboxamide 5 54a. 4-((Benzyloxy)carbonyl)-1-(tert-butoxycarbonyl)piperazine-2 carboxylic acid The tile compound was prepared according to the procedure of [Kempf, D. J.; Norbeck, D. W.; Sham, H. L. U.S. Patent 5,455,351, Oct 3, 1995]. Piperazine-2-carboxylic acid (10.0 g, 77.0 mmol) was dissolved in a 1:1 10 solution of 1,4-dioxane:water (100 mL) at room temperature with vigorous stirring. The clear solution was adjusted to pH 11 by the addition of an aqueous solution of sodium hydroxide (80 mL of a 1N solution). The pH was monitored in situ with a pH meter throughout the reaction. The reaction flask was fitted with an addition funnel that contained a solution of N-a 15 (benzyloxycarbonyloxy) succinamide (13.6 g, 55 mmol) in 1,4-dioxane (50 mL). The N-a-(benzyloxycarbonyloxy) succinamide solution was added over 45 minutes at room temperature and the pH was kept above 10 by the periodic addition of 1 N sodium hydroxide. The pH of the solution was adjusted to 9.5 and 2-(tert-butoxycarbonyloxyimino)-2-phenylacetonitrile (13.4 g, 55 20 mmol) was added as a solution in 1,4-dioxane (50 mL) over 10 minutes. The pH was maintained at 9.5 and the solution was stirred at room temperature for 17 hours. The solution was then acidified to pH 2 and the aqueous solution was washed with diethyl ether (3 x 150 mL). The aqueous solution was cooled to OC and acidified by adding of concentrated hydrochloric acid. The acidic 25 solution was extracted with ethyl acetate (5 x 150 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was triturated with a 1:1 solution of dichloromethane: hexanes (150 mL) and the solvent was removed in vacuo to provide the product as a viscous yellow oil (15.7 g, 43 mmol, 80%). Rf = 0.60 (66:34 dichloromethane: 30 ethyl acetate + 0.1% (v/v acetic acid); 1 H-NMR (400 MHz, DMSO) 6 13.0 (br s, 1H), 7.37-7.36 (m, 5H), 5.05 (s, 2H), 4.54-4.33 (m, 2H), 3.90-3.66 (m, 2H), 3.07-2.81 (m, 4H), 1.38 (s, 9H); Mass spectrum (ESI +ve) m/z 365.1 (MH*).

118 54b. 4-Benzyl 1-tert-butyl 2-(methylcarbamoyl)piperazine-1,4 dicarboxylate 4-((Benzyloxy)carbonyl)-1 -(tert-butoxycarbonyl)piperazine-2-carboxylic acid (1.70 g, 4.70 mmol), DMF (20 mL), diisopropylethylamine (2.50 mL, 14.1 5 mmol) and methylamine hydrochloride (0.350 g, 5.20 mmol) were mixed together at room temperature under argon for 10 minutes. 2-(7-aza-1H benzotriazole-1-yI)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU, 2.00 g, 5.20 mmol) was then added to the reaction mixture in one portion. The mixture was stirred at room temperature under argon for 18 hours. The 10 reaction mixture was then poured into water (100 mL) and extracted with ethyl acetate (4 x 25 mL). The combined organic phases were washed with saturated ammonium chloride (3 x 15 mL), water (3 x 15 mL) and brine (70 mL). The combined organic phases were then dried over sodium sulfate, filtered and concentrated in vacuo. The product, obtained as a white foam 15 (0.91 g, 2.4 mmol, 51%) was purified by column chromatography (gradient elution 20:80 ethyl acetate: hexanes to 100% ethyl acetate). Rf = 0.10 (50:50 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.39-7.33 (m, 5H), 5.17 (br s, 2H), 4.68-4.58 (m, 2H), 3.98-3.88 (m, 2H), 3.23-3.08 (m, 4H), 2.07 (s, 3H), 1.50 (s, 9H); Mass spectrum (ESI +ve) m/z 378.0 (MH*). 20 54c. tert-Butyl 2-(methylcarbamoyl)piperazine-1 -carboxylate 4-Benzyl 1-tert-butyl 2-(methylcarbamoyl)piperazine-1,4-dicarboxylate (0.910 g, 2.40 mmol) was dissolved in methanol (10 mL) at room temperature with stirring and the vial was flushed with argon. Palladium on carbon (91.0 mg of 1Owt% on carbon) was added in one portion to the stirred reaction 25 mixture. The reaction flask was charged with hydrogen gas (1 atm) and stirred for 18 hours at room temperature. The palladium on carbon was removed by vacuum filtration through Celite and rinsed with additional methanol (5 x 10 mL). The combined filtrates were concentrated in vacuo. The product, obtained as a yellow solid (0.16 g, 0.66 mmol, 27%) was purified by column 30 chromatography (isocratic 3:97 methanol: dichloromethane + 0.1% (v/v) ammonium hydroxide). Rf = 0.32 (3:97 methanol: dichloromethane + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDC 3 ) 6 6.41 (br s, 1H), 119 4.61-4.59 (m, 1H), 3.68-3.65 (m, 1H), 3.18-2.85 (m, 6H), 2.47 (br s, 3H), 1.51 (s, 9H); Mass spectrum (ESI +ve) m/z 243.9 (MH*). 54d. (E)-tert-Butyl 2-(methylcarbamoyl)-4-(3-(2,6,6-trimethylcyclohex-l 5 en-1-yl)acryloyl) piperazine-1-carboxylate (E)-3-(2,6,6-trimethylcyclohex-1-en-1-yl)acrylic acid (0.120 g, 0.620 mmol), diisopropylethylamine (0.210 mL, 1.20 mmol), tert-butyl 2 (methylcarbamoyl)piperazine-1-carboxylate (0.150 g, 0.620 mmol) were dissolved in a 1:5 mixture of dichloromethane: acetonitrile at room 10 temperature under argon. 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate (HATU, 0.23 g, 0.62 mmol) was added in one portion and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was passed through a carbonate SPE cartridge (silica-carbonate Silicycle, 2 g, 230-400 mesh) followed by filtration 15 through a tosic acid SPE cartridge (silica-tosic acid Silicycle, 1 g, 230-400 mesh) and the solvent was removed in vacuo. The product, obtained as a clear oil (0.17 g, 0.41 mmol, 66%) was purified by column chromatography (gradient elution 30:70 ethyl acetate: hexanes to 100 % ethyl acetate). The proton NMR spectrum shows evidence that the product is a mixture of 20 rotamers. Rf = 0.14 (50:50 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.15-7.12 (m, 1H), 6.23-5.98 (m, 1H), 4.60-4.27 (m, 1H), 4.23-4.10 (m, 2H), 3.98-3.87 (m, 2H), 3.82-3.62 (m, 1H), 3.37-3.13 (m, 2H), 3.09-2.71 (m, 1H), 2.59 (s, 2H), 1.87 (s, 3H), 1.61-1.58 (m, 2H), 1.45-1.42 (m, 2H), 1.30 (s, 9H), 0.88 (s, 6H); Mass spectrum (ESI +ve) m/z 420.1 (MH*). 25 54e. (E)-N 2 -methyl-4-(3-(2,6,6-trimethylcyclohex-1 -en-I -yl)acryloyl) piperazine-1,2-dicarboxamide (E)-tert-Butyl 2-(methylcarbamoyl)-4-(3-(2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl)piperazine-1-carboxylate (0.170 g, 0.410 mmol) was dissolved in dichloromethane (5 mL) at room temperature. Trifluoroacetic acid (3 mL) was 30 added to the stirred solution and the reaction mixture was stirred at room temperature for 1 hour and then concentrated in vacuo to provide the crude 120 trifluoroacetic acid salt as a viscous yellow oil (0.42 g). The trifluoroacetic acid salt was analyzed by thin layer chromatography and LC-MS then used directly in the next reaction. Rf = 0.17 (7:93 methanol: dichloromethane + 0.1% (v/v) ammonium hydroxide, ninhydrin staining); Mass spectrum (ESI +ve) m/z 5 320.1 (MH*). The trifluoroacetic acid salt (0.4 g) and potassium carbonate (0.54 g, 3.9 mmol) were stirred at room temperature under argon for 20 minutes. Trimethylsilyl isocyanate (0.32 mL, 3.9 mmol) was added in one portion and the mixture was stirred for 18 hours at room temperature. The reaction 10 mixture was poured into saturated ammonium chloride (15 mL) and extracted with dichloromethane (3 x 10 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The product, obtained as a white solid (45 mg, 0.12 mmol, 32%) was purified by preparative thin layer chromatography (1000 pm thickness Si02 gel, 20 cm x 20 cm plate, 15 eluent 10:90 methanol: dichloromethane + 0.1% (v/v) ammonium hydroxide). The proton NMR spectrum shows evidence that the product is a mixture of rotamers. Mp = 100-112*C; Rf = 0.62 (10:90 methanol: dichloromethane + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.43-7.34 (m, 1H), 6.47-6.40 (m, 1H), 5.18-4.61 (m, 4H), 4.35-4.15 (m, 1H), 3.93-3.67 (m, 20 1H), 3.46-2.97 (m, 3H), 2.83-2.82 (m, 3H), 2.10-2.06 (m, 2H), 1.80 (s, 3H), 1.65-1.61 (m, 2H), 1.51-1.48 (m, 2H), 1.09 (m, 6H); Mass spectrum (ESI +ve) m/z 363.1 (MH*). Example 55: Nl-((2,6,6-Trimethylcyclohex-1 -en-1 -yI)methyl)piperazine 25 1,4-dicarboxamide 55a. tert-Butyl 4-(((2,6,6-Trimethylcyclohex-1-en-1-yl)methyl)carbamoyl) piperazine-1 -carboxylate To a stirred solution of 2-(2,6,6-trimethylcyclohex-1-en-1-yl)acetic acid (0.30 g, 1.6 mmol) in anhydrous benzene (16 mL) was added 30 diphenylphosphoryl azide (0.45 g, 1.6 mmol) and triethylamine (0.51 g, 4.8 mmol). The pale yellow reaction mixture was heated to reflux for 3 hours until 121 it turned a blue color. The reaction was cooled to room temperature and ter butyl piperazine-1-carboxylate (0.31 g, 1.6 mmol) and triethylamine (0.17 g, 1.6 mmol) were added to the reaction mixture. The reaction was stirred at room temperature for 18 hours overnight. 5 The reaction mixture was diluted with ethyl acetate (150 mL), transferred to a separatory funnel and extracted with a 50% saturated solution of ammonium chloride (2 x 75 mL), and a saturated solution of sodium bicarbonate (2 x 75 mL). the organic layer was then washed with brine (75 mL), dried over magnesium sulfate, filtered and concentrated in vacuo to give 10 a pale yellow solid (0.60 g, quantitative). Rf = 0.65 (25:75 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 8 4.00 (s, 1H), 3.79 (d, J = 4.0 Hz, 2H), 3.52-3.17 (m, 8H), 2.04-1.82 (m, 2H), 1.63 (s, 3H), 1.60-1.54 (m, 2H), 1.44 (s, 11 H), 0.98 (s, 6H); Mass spectrum (ESI +ve) m/z 366 (MH*). 55b. N-((2,6,6-Trimethylcyclohex-1-en-1-yl)methyl)piperazine-1 15 carboxamide The title compound, obtained as a yellow oil (265 mg, quantitative) was prepared from the product of Example 55a by following the procedure of example 49b. The crude product was carried forward without purification. Rf= 0.05 (95:5 chloroform: methanol); Mass spectrum (ESI +ve) m/z 266 (MH*). 20 55c. N'-((2,6,6-Trimethylcyclohex-1-en-1-yl)methyl)piperazine-1,4 dicarboxamide The title compound, obtained as a pale yellow solid (33 mg, 12%) was prepared from the product of Example 55b following the procedure of 25 Example 33. Mp = 168.9-169.7 0 C. Rf = 0.45 (90:10 chloroform: methanol); 1 H NMR (400 MHz, d 4 -MeOD) 8 3.79-3.68 (m, 3H), 3.27 (s, 3H), 1.96 (t, J = 6.0 Hz, 2H), 1.77-1.56 (m, 6H), 1.48-1.41 (m, 2H), 1.35-1.08 (m, 1H), 0.99 (s, 6H), 0.93 (d, J = 6.5 Hz, 1H), 0.88 (d, J = 6.5 Hz, 1H), 0.06 (d, J = 4.0 Hz, 1H); Mass spectrum (ESI +ve) m/z 308 (MH*). 30 122 Example 56: N'-Methyl-N1-((2,6,6-trimethycy clohex-1 -en-1 -yl)methyl) piperazine-1,4-dicarboxamide 56a. N-Methyl-N-((2,6,6-trimethylcyclohex-1-en-1-yl)methyl)-1H 5 imidazole-1-carboxamide N-Methyl-1-(2,6,6-cyclohex-1-en-1-yl)methanamine hydrochloride (1.00 g, 4.90 mmol) and triethylamine (0.750 mL, 5.40 mmol) were stirred under argon at room temperature in THF (12 mL) for 15 minutes. Carbonyldiimidazole (0.88 g, 5.4 mmol) was added in one portion to the 10 stirred reaction mixture and the reaction mixture was heated to reflux for 18 hours. The solvent was removed in vacuo and the residue was dissolved in dichloromethane (100 mL). The organic phase was washed with water (2 x 75 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford the product as a yellow oil (1.2 g, 4.6 mmol, 93%). Rf = 0.90 (7:93 methanol: 15 dichloromethane + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.90 (s, 1H), 7.24 (s, 1H), 7.10 (s, 1H), 4.26 (s, 2H), 2.94 (s, 3H), 2.07-2.04 (m, 2H), 1.72 (s, 3H), 1.68-1.62 (m, 2H), 1.50-1.47 (m, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 261.9 (MH*). 56b. 3-Methyl-1-(methyl((2,6,6-trimethylcyclohex-1-en-1 20 yI))methyl)carbamoyl)-1H-imidazol-3-ium iodide N-Methyl-N-((2,6,6-trimethylcyclohex-1 -en-1 -yl)methyl)-i H-imidazole-1 carboxamide (1.20 g, 4.60 mmol) and iodomethane (1.1 mL, 18 mmol) were dissolved in acetonitrile and stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo to provide the product as a 25 hygroscopic yellow foam (1.8 g, 4.4 mmol, 97%). 1 H-NMR (400 MHz, DMSO) 8 9.63 (br s, 1H), 8.07 (br s, 1H), 7.87 (s, 1H), 4.24 (br s, 2H), 3.91 (s, 3H), 2.88 (s, 3H), 2.10-2.08 (m, 2H), 1.70-1.59 (m, 5H), 1.46-1.44 (m, 2H), 1.03 0.95 (m, 6H); Mass spectrum (ESI +ve) m/z 275.9 (MH*). 30 123 56c. N'-Methyl-N'-((2,6,6-trimethylcyclohex-1-en-1-yl)methyl)piperazine 1,4-dicarboxamide 3-Methyl-1 -(methyl((2,6,6-trimethylcyclohex-1 -en-1 yl))methyl)carbamoyl)-1H-imidazol-3-ium iodide (0.200 g, 0.470 mmol), 5 piperazine-1-carboxamide hydrochloride (78.0 mg, 0.470 mmol) and triethylamine (0.130 mL, 0.930 mmol) were dissolved in a 1:4 mixture of acetonitrile: dichloromethane. The reaction mixture was stirred at room temperature under argon for 2 days. The reaction mixture was poured into saturated ammonium chloride (30 mL) and the organic layer was removed. 10 The aqueous layer was extracted with dichloromethane (4 x 15 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The product obtained as a white solid (11 mg, 0.03 mmol, 7%) was purified by preparative plate thin layer chromatography (1000 pm thickness SiO 2 gel, 20 cm x 20 cm plate, eluent 10:90 methanol: ethyl 15 acetate + 0.1% (v/v) ammonium hydroxide. Mp = 139.6-140.3*C; Rf = 0.62 (10:90 methanol: ethyl acetate + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, DMSO) 5 6.00 (s, 2H), 3.93 (s, 2H), 3.32-3.28 (m, 4H), 3.01-3.00 (m, 4H), 2.66 (s, 3H), 1.99-1.96 (m, 2H), 1.65 (s, 3H), 1.59-1.57 (m, 2H), 1.41 1.40 (m, 2H), 0.96 (s, 6H); Mass spectrum (ESI +ve) m/z 323.0 (MH*). 20 Example 57: (RE)-1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl) pyrrolidin-3-yi carbamate 57a. (RE)-1-(3-Hydroxypyrrolidin-1-yI)-3-(2,6,6-trimethylcyclohex-1-en-1 25 yl)prop-2-en-1-one The title compound, obtained as a colorless oil (0.35 g, 86%), was prepared by following the procedure of Example 3, except (R)-pyrrolidin-3-ol was substituted for methylamine hydrochloride. [aDo 23 = -13.78* (c = 0.005, methanol); Rf = 0.2 (100% ethyl acetate); 1 H-NMR (400 MHz, CDCl 3 ) 5 7.36 30 (d, J = 15.5 Hz, 1H), 6.09 (d, J = 15.5 Hz, 1H), 4.55 (m, 1H), 3.80-3.53 (m, 124 4H), 2.95 (m, 1H), 2.12-1.97 (m, 4H), 1.76 (s, 3H), 1.63 (m, 2H), 1.48 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 264 (MH*). 57b. (RE)-1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl) pyrrolidin-3-yi carbamate 5 Trichloroacetyl isocyanate (501 mg, 2.66 mmol) was added to a solution of 57a (350 mg, 1.33 mmol) in tetrahydrofuran (3 mL). The solution was stirred for 12 hours at room temperature and then treated with water (0.5 mL) to destroy the excess of trichloroacetyl isocyanate. The reaction mixture was diluted with ethyl acetate (50 mL) and extracted with water (30 mL). The 10 organic layer was separated, dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was carried forward without further purification. A solution of potassium carbonate (367 mg, 2.66 mmol) in water (8 mL) was added to a solution of the crude material (600 mg, 1.33 mmol) in 15 terahydrofuran (10 mL) and methanol (10 mL). The mixture was stirred for 3 hours at room temperature, and then diluted with ethyl acetate (60 mL) and water (60 mL). The organic layer was separated, dried over magnesium sulfate, filtered and concentrated in vacuo to yield the title compound as an off-white solid (72 mg, 18%); [a]D 23 = -12.42* (c = 0.006, chloroform); Rf = 0.2 20 (10:90 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.41 (d, J = 15.5 Hz, 1H), 6.08 (d, J = 15.5 Hz, 1H), 5.31 (m, 1H), 4.72 (m, 2H), 3.88-3.77 (m, 2H), 3.77-3.62 (m, 2H), 2.18 (m, 2H), 2.05 (m, 2H), 1.77 (s, 3H), 1.64 (m, 2H), 1.49 (m, 2H), 1.08 (m, 6H); Mass spectrum (ESI +ve) m/z 307 (MH*). 25 125 Example 58: (S,E)-1-(1-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acryloyl) pyrrolidin-3-yl)urea 58a. (S,E)-tert-Butyl (1-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acryloyl) 5 pyrrolidin-3-yi) carbamate The title compound, obtained as a colorless oil (62 mg, 35%), was prepared by following the procedure of Example 3, except (S)-tert-butyl pyrrolidin-3-yl carbamate was substituted for methylamine hydrochloride. Rf = 0.15 (100% ethyl acetate); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.39 (dd, J = 15.5, 10 6.0 Hz, 1H), 6.14-6.00 (m, 1H), 4.28 (m, 1H), 3.81 (m, 2H), 3.73-3.32 (m, 4H), 2.32-2.12 (m, 1H), 2.06 (s, 3H), 1.83 (d, J = 6.5 Hz, 1H), 1.79 (s, 3H), 1.65 (m, 4H), 1.48 (m, 9H), 1.07 (m, 6H); Mass spectrum (ESI +ve) m/z 363 (MH*). 58b. (S,E)-1-(3-Aminopyrrolidin-1-yI)-3-(2,6,6-trimethylcyclohex-1-en-1 15 yl)prop-2-en-1 -one The title compound, obtained as white solid (42 mg, 98%), was prepared from the product of Example 58a by following the procedure of Example 49b. Rf = 0.05 (10:89:1 methanol: dichloromethane: ammonium hydroxide); 1 H NMR (400 MHz, DMSO-d 6 ) 8 8.40 (br s, 3H), 7.15 (d, J = 16.0 20 Hz, 1H), 6.24-6.09 (m, 1H), 3.95-3.51 (m, 4H), 2.33-2.19 (m, 1H), 2.04 (m, 4H), 1.73 (s, 3H), 1.65-1.53 (m, 2H), 1.51-1.38 (m, 2H), 1.03 (s, 3H),- 1.01 (s, 3H); Mass spectrum (ESI +ve) m/z 263 (MH*). 58c. (S,E)-1-(1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl)pyrrolidin-3 yl)urea 25 The title compound, obtained as white film (6.7 mg, 12%), was prepared from the product of Example 58b by following the procedure of Example 57b. Rf = 0.2 (5:95 methanol: chloroform); 1 H NMR (400 MHz,) 8 7.13 (d, J = 15.5 Hz, 1H), 6.31 (m, 1H), 6.14 (d, J = 15.5 Hz, 1H), 5.45 (m, 2H), 4.08 (m, 1H), 3.58 (t, J = 7.0 Hz, 1H), 3.53-3.14 (m, 4H), 2.10-1.95 (m, 30 2H), 1.73 (s, 3H), 1.58 (m, 2H), 1.46 (m, 2H), 1.03 (m, 6H); Mass spectrum (ESI +ve) m/z 306 (MH*).

126 Example 59: (RE)-1 -(1 -(3-(2,6,6-trimethylcyclohex-1 -en-1 -yl)acryloyl) pyrrolidin-3-yl)urea 59a. (RE)-tert-Butyl (1 -(3-(2,6,6-trimethylcyclohex-1 -en-1 -yl)acryloyl) 5 pyrrolidin-3-yl) carbamate The title compound, obtained as a colorless oil (300 mg, 65%), was prepared by following the procedure of Example 3, except (R)-tert-butyl pyrrolidin-3-ylcarbamate was substituted for methylamine hydrochloride. Rf = 0.15 in (40:60 ethyl acetate:hexane); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.39 (dd, J 10 = 15.5, 6.0 Hz, 1H), 6.14-6.00 (m, 1H), 4.28 (m, 1H), 3.81 (m, 2H), 3.73-3.32 (m, 4H), 2.32-2.12 (m, 1H), 2.06 (s, 3H), 1.83 (d, J = 6.5 Hz, 1H), 1.79 (s, 3H), 1.65 (m, 4H), 1.48 (m, 9H), 1.07 (m, 6H); Mass spectrum (ESI +ve) m/z 363 (MH*). 59b. (RE)-1 -(3-Aminopyrrolidin-1 -yI)-3-(2,6,6-trimethylcyclohex-1 -en-1 15 yl)prop-2-en-1-one The title compound, obtained as white solid (70 mg, 84%), was prepared from the product of Example 59b by following the procedure of Example 49b. Rf = 0.05 (10:89:1 methanol: dichloromethane: ammonium hydroxide); 1 H NMR (400 MHz, DMSO-d 6 ) 8 8.4 (br s, 3H), 7.15 (d, J = 16.0 20 Hz, 1H), 6.24-6.09 (m, 1H), 3.95-3.51 (m, 4H), 2.33-2.19 (m, 1H), 2.04 (m, 4H), 1.73 (s, 3H), 1.65-1.53 (m, 2H), 1.51-1.38 (m, 2H), 1.01 (m, 6H); Mass spectrum (ESI +ve) m/z 263 (MH*). 59c. (RE)-1-(1-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl) pyrrolidin 3-yl)urea 25 The title compound, obtained as white film (21 mg, 10%), was prepared from the product of Example 59b by following the procedure of Example 57b: Rf = 0.2 (5:95 methanol: chloroform); 'H NMR (400 MHz, DMSO-d 6 ) 8 7.13 (d, J = 15.5 Hz, 1H), 6.31 (m, 1H), 6.14 (d, J = 15.5 Hz, 1H), 5.45 (m, 2H), 4.08 (m, 1H), 3.58 (t, J = 7.0 Hz, 1H), 3.53-3.14 (m, 4H), 2.10-1.95 (m, 2H), 1.73 30 (s, 3H), 1.58 (m, 2H), 1.46 (m, 2H), 1.03 (m, 6H); Mass spectrum (ESI +ve) m/z 306 (MH*).

127 Example 60: (E)-4-(3-(2,6,6-Trimethyl-3-oxocyclohex-I-en-1 yl)acryloyl)piperazine-I -carboxamide 60a. (E)-Methyl 3-(2,6,6-trimethylcyclohex-I-enyl)acrylate 5 (E)-3-(2,6,6-Trimethylcyclohex-1-enyl)acrylic acid (5.10 g, 26.3 mmol) was dissolved in acetone (20 mL) and anhydrous potassium carbonate (3.66 g, 26.5 mmol) was added and the reaction mixture was stirred vigorously. Methyl iodide (4.11 g, 1.80 mL, 28.9 mmol) was added via syringe and the reaction mixture was stirred at room temperature for 3 days. 10 The reaction was dissolved in diluted with diethyl ether (175 mL) then extracted with distilled water (100 mL), saturated sodium bicarbonate (100 mL) and brine (100 mL). The combined aqueous layers were extracted with diethyl ether (2 x 75 mL) and the combined organic layers were dried over sodium sulfate and concentrated in vacuo to yield a viscous yellow oil (5.17 15 g). The product was purified by flash chromatography to yield a clear oil (3.77 g, 70%). Rf = 0.21 (20:80 ethyl acetate:hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.42 (d, J = 16.0 Hz, 1H), 5.80 (d, J = 16.0 Hz, 1H), 3.74 (s, 3H), 2.03 (t, J = 6.4 Hz, 2H), 1.72 (s, 3H), 1.61-1.58 (m, 2H), 1.47-1.44 (m, 2H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 209 (MH*). 20 60b. (E)-Methyl 3-(2,6,6-trimethyl-3-oxocyclohex-I-enyl)acrylate (E)-Methyl 3-(2,6,6-trimethylcyclohex-1-enyl)acrylate (3.77 g, 18.0 mmol) was dissolved in 1,4-dioxane (60 mL) to which selenium dioxide (2.00 g, 18.0 mmol) was added and allowed to stir vigorously. The reaction mixture was sealed with a rubber septum and placed into an 800C oil bath for 16 25 hours. The reaction was filtered and concentrated in vacuo to yield a brown oil (5.20 g). The product was purified by flash column chromatography to yield a yellow oil (440 mg, 11%). Rf = 0.3 (10:90 ethyl acetate:hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.41 (d, J = 16.4 Hz, 1H), 5.92 (d, J =16.4 Hz, 1H), 3.80 30 (s, 3H), 2.51 (d, J= 6.8 Hz, 2H), 1.88 (d, J= 7.2 Hz, 2H), 1.80 (s, 3H), 1.18 (s, 6H); Mass spectrum (ESI +ve) m/z 223 (MH*).

128 60c. (E)-3-(2,6,6-Trimethyl-3-oxocyclohex-1-enyl)acrylic acid (E)-Methyl 3-(2,6,6-trimethyl-3-oxocyclohex- 1 -enyl)acrylate (830 mg, 4.50 mmol) was dissolved in tetrahydrofuran (30 mL) at room temperature under argon to which a solution of lithium hydroxide (210 mg, 5.00 mmol) in 5 water (5 mL) was added. The reaction was stirred vigorously at room temperature under argon for 2 hours. The reaction was acidified with 1M hydrochloric acid solution at 0*C, diluted with water (150 mL) and extracted with ethyl acetate (3 x 85 mL). The combined organic layers were washed with brine (120 mL) and dried over 10 sodium sulfate. The solvent concentrated in vacuo to yield a yellow oil (427 mg, 55%). 'H-NMR (400 MHz, CDCl 3 ) 6 7.49 (d, J = 16.0 Hz, 1H), 5.95 (d, J =16.4 Hz, 1H), 2.54 (t, J = 6.8 Hz, 2H), 1.89 (t, J = 6.8 Hz, 2H), 1.81 (s, 3H), 1.19 (s, 6H); Mass spectrum (ESI +ve) m/z 209 (MH*). 60d. (E)-tert-Butyl 4-(3-(2,6,6-trimethyl-3-oxocyclohex-1-en-1 15 yl)acryloyl)piperazine-1 -carboxylate The title compound, obtained as a clear oil (60.0 mg, 26%), was prepared from the product of Example 60c by following the procedure of Example 3. Rf = 0.25 (50:50 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDC1 3 ) 6 7.38 (d, J = 15.6, 1H), 6.32 (d, J = 15.6 Hz, 1H), 3.97-3.48 (m, 8H), 20 2.52 (t, J = 6.8 Hz, 2H), 1.88 (t, J = 6.8 Hz, 2H), 1.81 (s, 3H), 1.41 (s, 9H), 1.18 (s, 6H); Mass spectrum (ESI +ve) m/z 377 (MH*). 60e. (E)-4-(3-(2,6,6-Trimethyl-3-oxocyclohex-1-en-1 yl)acryloyl)piperazine-1 -carboxamide The product of example 60d dissolved in dichloromethane and a 4.0 M 25 solution of hydrochloric acid in 1,4-dioxane was added to the stirred reaction mixture. The reaction was stirred at room temperature for 4 hours and then concentrated in vacuo to afford a pale yellow crude oil. The title compound, obtained as a white film (23.0 mg, 45%), was prepared from the product of Example 60d by following the procedure of Example 33. Rf = 0.50 (10:90 30 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.39 (d, J = 15.6 Hz, 1H), 6.32 (d, J = 15.6 Hz, 1H), 4.64 (s, 2H), 3.76-3.45 (m, 8H), 2.52 (t, J = 6.8 Hz, 129 2H), 1.88 (t, J = 6.8 Hz, 2H), 1.80 (s, 3H), 1.18 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 319.9 (MH*). Example 61: (E)-4-(3-(3-Hydroxy-2,6,6-trimethylcyclohex-1-en-1-yl) 5 acryloyl)piperazine-1-carboxamide 61a. (E)-2,4,4-Trimethyl-3-(3-oxobut-1-en-1-yl)cyclohex-2-en-1-yI acetate To a solution of 1,4-benzoquinone (6.00 g, 55.5 mmol) and beta-ionone (10.6 g, 55.5 mmol) in acetic acid (180 mL) were added palladium 10 bis(trifluoroacetate) (900 mg, 3.00 mmol) and o-methoxyacetophenone (1.68 g, 11.1 mmol). The mixture was heated to 70 0 C for 12 hours. The solvent was concentrated in vacuo and then a solution of sodium hydroxide (200 mL, 6 N) was added, and the aqueous phase extracted with diethyl ether (5 x 50 mL). The combined organic extracts were washed with a saturated solution of 15 sodium carbonate (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (90:10 hexane: diethyl ether) to yield the title compound as a brown oil (7.8 g; 56%). Rf = 0.25 (30:70 diethyl ether: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 5 7.20 (d, J = 16.5 Hz, 1H), 6.15 (d, J = 16.5 Hz, 1H), 5.25 (m, 20 1H), 2.33 (s, 3H), 2.10 (s, 3H), 2.00-1.88 (m, 1H), 1.81-1.73 (m, 1H), 1.72 (s, 3H), 1.71-1.60 (m, 1H), 1.48 (m, 1H), 1.09 (s, 3H), 1.04(s, 3H); Mass spectrum (ESI +ve) m/z 251 (MH*). 61b. (E)-3-(3-Hydroxy-2,6,6-trimethylcyclohex-1-en-1-yl)acrylic acid The title compound, obtained as clear yellow oil, was prepared from the 25 product of Example 61a by following the procedure of Example 1, except (E) 2,4,4-trimethyl-3-(3-oxobut-1 -en-1 -yl)cyclohex-2-en-1 -yl acetate was substituted for beta-ionone. 1 H NMR (400 MHz, CDCl 3 ) 5 7.51-7.45 (m, 1H), 5.89 (d, J = 16.0 Hz, 1H), 4.06 (t, J = 4.5 Hz, 1H), 1.99-1.90 (m, 2H), 1.90 (s, 3H), 1.80-1.63 (m, 2H), 1.47 (m, 1H), 1.10 (s, 3H), 1.07 (s, 3H).

130 61 c. (E)-tert-Butyl 4-(3-(3-hydroxy-2,6,6-trimethylcyclohex-1 -en-1 yl)acryloyl)piperazine-1 -carboxylate The title compound, obtained as a colorless oil (50 mg, 98%), was prepared by following the procedure of Example 3, except tert-butyl 5 piperazine-1-carboxylate was substituted for methylamine hydrochloride and (E)-3-(3-hydroxy-2,6,6-trimethylcyclohex-1-en-1-yl)acrylic acid was substituted for (E)-3-(2,6,6-trimethylcyclohex-1-en-1-yl)acrylic acid. Rf = 0.10 (50:50 ethyl acetate: hexanes); 'H-NMR (400 MHz, CDC 3 ) 8 7.33 (s, 1H), 6.25 (d, J = 15.5 Hz, 1H), 4.03 (t, J = 4.5 Hz, 1H), 3.66 (m, 2H), 3.62-3.51 (m, 2H), 3.49 (m, 10 4H), 2.00-1.89 (m, 1H), 1.88 (s, 3H), 1.78-1.63 (m, 4H), 1.53-1.42 (m, 9H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 379 (MH*). 61 d. (E)-4-(3-(3-Hydroxy-2,6,6-trimethylcyclohex-1 -en-1 -yl) acryloyl) piperazine-1 -carboxamide The product of example 61c dissolved in dichloromethane and a 4.0 M 15 solution of hydrochloric acid in 1,4-dioxane was added to the stirred reaction mixture. The reaction was stirred at room temperature for 4 hours and then concentrated in vacuo to afford a pale yellow crude oil. The title compound, obtained as colorless oil (8 mg, 4%), was prepared from the product of Example 61c by following the procedure of Example 33. Rf = 0.15 (5:95 20 methanol: chloroform); 'H NMR (400 MHz, DMSO-d 6 ) 8 7.34 (d, J = 15.5 Hz, 1H), 6.25 (d, J = 15.5 Hz, 1H), 5.81-5.53 (m, 2H), 4.05 (m, 1H), 3.81 (m, 2H), 3.65 (m, 2H), 3.54 (m, 4H), 1.98 (m, 1H), 1.87 (s, 3H), 1.80-1.65 (m, 2H), 1.53-1.43 (m, 1H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 322 (MH*). 25 Example 62: (E)-4-(3-(3,3-Difluoro-2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl)piperazine-1-carboxamide 62a. (E)-Methyl 3-(6,8,8-trimethyl-1,4-dithiaspiro[4.5]dec-6-en-7 yl)acrylate (E)-Methyl 3-(2,6,6-trimethyl-3-oxocyclohex-1-en-1-yl)acrylate (0.550 g, 30 2.40 mmol) was dissolved in 1,2-ethanedithiol (0.710 mL, 8.50 mmol) at room temperature under argon. The homogenous mixture was then cooled to -15 0

C

131 and stirred for 10 minutes. Zinc(II)chloride (17.0 mg, 0.120 mmol) was added in one portion and the mixture was stirred at -15*C for 3 hous and then at room temperature for 18 hours. The reaction mixture was then diluted with water (30 mL). The aqueous layer was extracted with ethyl acetate (3 x 20 5 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The product, obtained as white crystals (0.6 g, 2.0 mmol, 85%) was purified by column chromatography (isocratic 5% ethyl acetate: hexanes). Mp = 79.5-88.1 0 C; Rf = 0.70 in (15:85 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.36 (d, J = 16.2 Hz, 1H), 5.87 (d, J = 10 16.2 Hz, 1H), 3.79 (s, 3H), 3.42-3.31 (m, 4H), 2.32-2.25 (m, 2H), 2.00 (s, 3H), 1.75-1.69 (m, 2H), 1.58 (s, 2H), 1.06 (s, 6H); Mass spectrum (ESI +ve) m/z 298.8. 62b. (E)-Methyl 3-(3,3-difluoro-2,6,6-trimethylcyclohex-1 -en-I -yl)acrylate A slurry of N-iodosuccinamide (1.12 g, 5.00 mmol) and 15 dichloromethane (6 mL) in a Nalgene bottle was cooled to -78 0 C under argon. Hydrofluoric acid-pyridine complex (1.40 mL, 49.6 mmol) was slowly added to the slurry and stirred for 10 minutes under argon. A solution of (E)-Methyl 3 (6,8,8-trimethyl-1,4-dithiaspiro[4.5]dec-6-en-7-yl)acrylate (62a, 0.370 g, 1.20 mmol) in dichloromethane (1 mL) was added to the reaction mixture and 20 stirred for 1 hour at -78 0 C. The reaction mixture was poured into a 1:1 solution of saturated sodium sulfite: saturated sodium bicarbonate (100 mL). The aqueous solution was extracted with ethyl acetate (3 x 70 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The product, obtained as a bright yellow oil (0.16 g, 0.65 mmol, 51%) 25 was purified by column chromatography (isocratic 10% ethyl acetate: hexanes). Rf = 0.71 in (10:60 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.52 (d, J = 16.2 Hz, 1H), 5.90 (d, J = 16.2 Hz), 3.80 (s, 3H), 2.18 2.12 (m, 2H), 1.80 (s, 3H), 1.70-1.68 (m, 2H), 1.09 (s, 6H); 19 F-NMR (376 MHz, CDCl 3 ) 8 -93.6 (ddd, J = 3.0, 14.0, 14.0 Hz). 30 62c. (E)-3-(3,3-difluoro-2,6,6-trimethylcyclohex-1-en-1-yl)acrylic acid (E)-Methyl 3-(3,3-difluoro-2,6,6-trimethylcyclohex-1 -en-1 -yl)acrylate (62b, 57.0 mg, 0.230 mmol) was dissolved in a 3:1 solution of tetrahydrofuran: 132 water at room temperature under argon. A solution of lithium hydroxide (0.25 mL, 0.25 mmol, 1M aqueous solution) was added at room temperature and the reaction was stirred for 6 hours. The reaction mixture was poured into saturated ammonium chloride (20 mL) and extracted with dichloromethane (3 5 x 15 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. This provided the product as white crystals (54 mg, quantitative). Mp = 114.3-119.50C; Rf = 0.12 in (10:90 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.39 (d, J = 16 Hz, 1H), 5.91 (d, J = 16 Hz, 1H), 2.20-2.09 (m, 2H), 1.81 (s, 3H), 1.70-1.67 (m, 2H), 1.09 (s, 6H); 10 19 F-NMR (376 MHz, CDCl 3 ) 6 -94.1 (t, J = 14.0 Hz). 62d. (E)-(9H-fluoren-9-yl)methyl 4-(3-(3,3-difluoro-2,6,6 trimethylcyclohex-1-en-1-yl)acryloyl) piperazine-1 -carboxylate The title compound, obtained as a clear oil (0.11 g, 81%), was prepared from the product of Example 62c by following the procedure of 15 Example 3 except (9H-fluoren-9-yl)methyl piperazine-1-carboxylate hydrochloride was substituted for methylamine hydrochloride. Rf = 0.35 in (30:70 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.80 (d, J = 7.6 Hz, 2H), 7.58 (d, J = 7.2 Hz, 2H), 7.44 (t, J = 7.6 Hz, 2H), 7.35 (t, J = 7.6 Hz, 2H), 7.26 (d, J = 16 Hz, 1H), 6.28 (d, J = 16 Hz, 1H), 4.54 (d, J = 6 Hz, 2H), 20 4.27 (t, J = 6 Hz, 1H), 3.64-3.40 (m, 8H), 2.22-2.12 (m, 2H), 1.82 (s, 3H), 1.72-1.69 (m, 2H), 1.10 (s, 6H); 19 F-NMR (376 MHz, CDCl 3 ) 6 -93.8 (m); Mass spectrum (ESI +ve) m/z 521.1 (MH*). 62e. (E)-3-(3,3-difluoro-2,6,6-trimethylcyclohex-1-en-1-yI)-1-(piperazin-1 yl)prop-2-en-1 -one 25 (E)-(9H-fluoren-9-yl)methyl 4-(3-(3,3-difluoro-2,6,6-trimethylcyclohex-1 en-1-yl)acryloyl) piperazine-1-carboxylate (97.0 mg, 0.190 mmol), piperidine (0.4 mL, 20% v/v) and acetonitrile (2 mL) were stirred together at room temperature for 5 minutes. The reaction mixture was concentrated in vacuo and the residue was lyophilized for 18 hours. The product, obtained as a clear 30 oil (37 mg, 0.12 mmol, 67%) was purified by column chromatography (isocratic elution 5:95 methanol: dichloromethane + 0.1% (v/v) ammonium 133 hydroxide. Rf = 0.36 in (5:95 methanol: dichloromethane + 0.1% (v/v) ammonium hydroxide; 1 H-NMR (400 MHz, CDCI 3 ) 6 7.20 (d, J = 16Hz, 1H), 6.28 (d, J = 16 Hz), 3.71-3.68 (m, 2H), 3.59-3.49 (m, 2H), 2.95-2.86 (m, 4H), 2.19-2.09 (m, 2H), 1.80 (s, 3H), 1.69-1.66 (m, 2H), 1.07 s, 6H); 19 F-NMR (376 5 MHz, CDCl 3 ) 6 -93.3 (ddd, J = 3.3, 14.0, 14.0 Hz); Mass spectrum (ESI +ve) m/z 299.1 (MH*). 62f: (E)-4-(3-(3,3-Difluoro-2,6,6-trimethylcyclohex-1-en-1-yl)acryloyl) piperazine-1 -carboxamide 10 The title compound, obtained as white crystals (21 mg, 68%), was prepared from the product of Example 62e by following the procedure of Example 33. Mp = 181.3-182.4 0 C; Rf = 0.35 in (5:95 methanol: dichloromethane + 0.1% ammonium hydroxide; 1 H-NMR (400 MHz, CDCl 3 ) 6 7.26 (d, J = 16 Hz, 1H), 6.29 (d, J = 16 Hz, 1H), 3.80-3.40 (m, 8H), 2.24-2.10 15 (m, 2H), 1.95-1.86 (m, 2H), 1.81 (br s, 3H), 1.74-1.66 (m, 2H), 1.09 (s, 6H); 19 F-NMR (376 MHz, CDCI 3 ) 6 -93.8 (t, J = 14.0 Hz); Mass spectrum (ESI +ve) m/z 342.0 (MH*). Example 63: (E)-4-(3-(3,3-Dideutero-2,6,6-trimethylcyclohex-1 -en-1 20 yl)acryloyl) piperazine-1-carboxamide 63a. (E)-Methyl 3-(3-deutero-3-hydroxy-2,6,6-trimethylcyclohex-1-en-1 yl)acrylate To a stirred solution of (E)-methyl 3-(2,6,6-trimethyl-3-oxocyclohex-1 25 en-1-yl) acrylate (60b, 0.27 g, 1.2 mmol) in d 4 -methanol (6 mL) at 0 0 C was added sodium borodeuteride (0.051 g, 1.2 mmol) in one portion. The reaction was stirred at 0 0 C for 3 hours then quenched by adding a 50% saturated solution of ammonium chloride (25 mL) and ethyl acetate (50 mL). The biphasic reaction mixture was transferred to a separatory funnel and 30 separated, and the organic layer was further washed with water (25 mL) and brine (25 mL), and then dried over magnesium sulfate, filtered and 134 concentrated in vacuo to give a pale yellow oil (0.24 g, 88%). Rf = 0.33 (25:75 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 8 7.34 (d, J = 16.5 Hz, 1H), 5.84 (d, J = 16.5 Hz, 1H), 3.76 (s, 3H), 1.90 (ddd, J = 13.5, 11.0, 3.0 Hz, 1H), 1.84 (s, 3H), 1.72 (ddd, J = 13.5, 7.0, 2.0 Hz, 1H), 1.64 (m, 1H), 1.44 5 (ddd, J = 13.5, 7.0, 3.0 Hz, 1H), 1.23 (br s, 1H), 1.05 (s, 3H), 1.03 (s, 3H); Mass spectrum (ESI +ve) m/z 226 (MH*). 63b. (E)-Methyl 3-(3-acetoxy-3-deutero-2,6,6-trimethylcyclohex-1-en-1 yl)acrylate To a solution of (E)-methyl 3-(3-deutero-3-hydroxy-2,6,6 10 trimethylcyclohex-1-en-1-yI)acrylate (63a, 0.22 g, 0.98 mmol) in acetic anhydride (3 mL) was added N,N-dimethylaminopyridine (0.012 g, 0.098 mmol). The reaction was stirred at room temperature for 18 hours overnight. The reaction was diluted with ethyl acetate (25 mL) and transferred to a separatory funnel. The organic layer was extracted with a 1M solution of 15 sodium hydroxide (2 x 25 mL), washed with water (25 mL) and brine (25 mL), and then dried over magnesium sulfate, filtered and concentrated in vacuo to give a pale yellow oil (0.20 g, 78%). Rf = 0.45 (25:75 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 5 7.34 (d, J = 16.0 Hz, 1H), 5.85 (d, J = 16.0 Hz, 1H), 3.77 (s, 3H), 2.07 (s, 3H), 1.90 (ddd, J = 13.5, 11.0, 3.0 Hz, 1H), 1.73 20 (ddd, J = 10.0, 7.0, 3.5 Hz, 1H), 1.69 (s, 3H), 1.66-1.58 (m, 1H), 1.44 (ddd, J = 13.5, 7.0, 3.0 Hz, 1H), 1.07 (s, 3H), 1.03 (s, 3H); Mass spectrum (ESI +ve) m/z 208 (M-OAc*). 63c. (E)-Methyl 3-(3,3-dideutero-2,6,6-trimethylcyclohex-1-en-1 yl)acrylate 25 To a solution of (E)-methyl 3-(3-acetoxy-3-deutero-2,6,6 trimethylcyclohex-1-en-1-yl)acrylate (0.19 g, 0.71 mmol) in anhydrous tetrahydrofuran (18 mL) was added palladium tetrakistriphenylphosphine (0.55 g, 0.48 mmol) and sodium borodeuteride (0.13 g, 3.1 mmol). The reaction flask was sealed tightly to allow build up of pressure from the 30 liberated deuterium gas, and the reaction was stirred at room temperature for 18 hours overnight. The reaction was quenched by adding a 50% saturated solution of ammonium chloride (20 mL) and diethyl ether (50 mL). The 135 biphasic reaction mixture was filtered through Celite into a separatory funnel and separated, and the organic layer was further washed with a 50% saturated solution of ammonium chloride (50 mL) and brine (50 mL), then dried over magnesium sulfate, filtered and concentrated in vacuo to give a 5 pale yellow oil (0.10 g, 68%) which was carried forward without purification. Rf = 0.90 (25:75 ethyl acetate: hexanes). 63d. (E)-3-(3,3-Dideutero-2,6,6-trimethylcyclohex-1-en-1-yl)acrylic acid The title compound, obtained as a pale yellow solid (68 mg, 92%) was prepared from the product of Example 63c following the procedure of 10 Example 60c. The compound was carried forward without purification. Rf = 0.45 (25:75 ethyl acetate: hexanes). 63e. (E)-tert-Butyl 4-(3-(3,3-dideutero-2,6,6-trimethylcyclohex-1 -en-1 yl)acryloyl) piperazine-1-carboxylate The title compound, obtained as a pale yellow solid (126 mg, 15 quantitative) was prepared from the product of Example 63d following the procedure of Example 3 except tert-butyl piperazine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.55 (25:75 ethyl acetate: hexanes); Mass spectrum (ESI +ve) m/z 365 (MH*). 63f. (E)-3-(3,3-dideutero-2,6,6-trimethylcyclohex-1-en-1-yI)-1-(piperazin-1 20 yl) prop-2-en-1 -one The title compound, obtained as a yellow oil (51 mg, 56%) was prepared from the product of Example 63e by following the procedure of example 49b. The crude product was carried forward without purification. Rf = 0.30 (90:10 chloroform: methanol); Mass spectrum (ESI +ve) m/z 266 (MH*). 25 63g. (E)-4-(3-(3,3-dideutero-2,6,6-trimethylcyclohex-1 -en-1 -yl)acryloyl) piperazine-1 -carboxamide The title compound, obtained as an off-white solid (35 mg, 59%) was prepared from the product of Example 63f following the procedure of Example 33 except potassium carbonate was substituted for triethylamine. Mp = 148.4 30 149.7'C; Rf= 0.50 (90:10 chloroform: methanol); 1 H NMR (400 MHz, CDCl 3 ) 8 136 7.34 (d, J = 15.5 Hz, 1H), 6.17 (d, J = 15.5 Hz, 1H), 4.88 (s, 2H), 3.79-3.35 (m, 8H), 1.72 (s, 3H), 1.62-1.53 (m, 2H), 1.45 (dd, J = 7.5, 4.0 Hz, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 308 (MH*). 5 Example 64: (E)-N-(piperidin-4-yI)-3-(2,6,6-trimethylcyclohex-1-en-1 yl)acrylamide hydrochloride 64a. (E)-tert-Butyl 4-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acrylamido) piperidine-1 -carboxylate 10 The title compound, obtained as a clear oil (202 mg, 52%), was prepared from the product of Example la by following the procedure of Example 3 except tert-butyl 4-aminopiperidine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.20 (25:75 ethyl acetate: hexanes); 1

H

NMR (400 MHz, CDCl 3 ) 6 7.30 (d, J = 15.2, 1H), 5.70 (d, J = 15.2 Hz, 1H), 15 5.42-5.28 (m, 1H), 4.06-4.01 (m, 3H), 2.90-2.84 (m, 2H), 2.03-1.94 (m, 4H), 1.72 (s, 3H), 1.59 (t, J = 5.6 Hz, 2H), 1.58-1.47 (m, 11H), 1.34-1.31 (m, 2H), 0.99 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 376.9 (MH*). 64b. (E)-N-(Piperidin-4-yI)-3-(2,6,6-trimethylcyclohex-1 -en-1 -yl)acrylamide hydrochloride 20 The title compound, obtained as a yellow solid (166 mg, 99%), was prepared by following the procedure of Example 49b. Rf = 0.1 (90:10 dichloromethane: methanol + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.31 (d, J = 16.0, 1H), 5.99 (d, J = 16.0 Hz, 1H), 4.06-4.03 (m, 1H), 3.65 (s, 2H), 3.46-3.43 (m, 2H), 3.34-3.30 (m, 2H), 2.22-2.01 (m, 4H), 25 1.81-1.77 (m, 5H) 1.64-1.63 (m, 2H), 1.43-1.41 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 277 (MH*).

137 Example 65: (E)-N-Methyl-N-(piperidin-3-yI)-3-(2,6,6-trimethylcyclohex-1 en-1-yl)acrylamide hydrochloride 65a. (E)-tert-Butyl 3-(N-methyl-3-(2,6,6-trimethylcyclohex-1 -en-1 5 yl)acrylamido)piperidine-1-carboxylate The title compound, obtained as a yellow solid (335 mg, 83%), was prepared from the product of Example la by following the procedure of Example 3 except tert-butyl 3-(methylamino)piperid ine- 1-ca rboxylate was substituted for methylamine hydrochloride. Rf = 0.20 (25:75 ethyl acetate: 10 hexanes); 'H-NMR (400 MHz, CDC 3 ) 8 7.33 (d, J = 15.0, 1H), 6.50 (d, J = 15.0 Hz, 1H), 4.44-3.78 (m, 4H), 2.95 (s, 3H), 2.84-2.78 (m, 1H), 2.58-2.56 (m,1H), 2.04-2.02 (m, 2H), 1.75 (s, 4H), 1.63-1.60 (m, 4H), 1.48-1.44 (m, 11 H), 1.01 (s, 6H); Mass spectrum (ESI +ve) m/z 391 (MH*). 65b. (E)-N-Methyl-N-(piperidin-3-yI)-3-(2,6,6-trimethylcyclohex-1-en-1-yl) 15 acrylamide hydrochloride The title compound, obtained as a yellow oil (278 mg, 99%), was prepared following the procedure of Example 49b. Rf = 0.2 (90:10 dichloromethane: methanol + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.31 (d, J = 15.0, 1H), 6.34 (d, J = 15.0 Hz, 1H), 4.81-4.76 (m, 20 1H), 3.39-3.31 (m, 4H), 3.07-2.78 (m, 4H), 2.09 (t, J = 6.0 Hz, 3H), 2.05-1.82 (m, 3H), 1.79 (s, 3H), 1.53-1.49 (m, 2H), 1.28-1.24 (m, 2H), 1.08 (s, 6H); Mass spectrum (ESI +ve) m/z 291 (MH*). Example 66: (E)-N-(Piperidin-3-yl)-3-(2,6,6-trimethylcyclohex-1-en-1 25 yl)acrylamide hydrochloride 66a. (E)-tert-Butyl 3-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acrylamido) piperidine-1 -carboxylate The title compound, obtained as a yellow oil (103 mg, 27%), was 30 prepared from the product of Example la by following the procedure of 138 Example 3 except tert-butyl 3-aminopiperidine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.20 in (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.31 (d, J = 15.2, 1H), 5.69 (d, J = 15.2 Hz, 1H), 4.07 (s, 1H), 3.49-3.28 (m, 4H), 2.04-2.01 (m, 2H), 1.89 (s, 1H), 1.73 (s, 3H), 5 1.71-1.58 (m, 5H), 1.55-1.46 (m, 11H),1.05 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 377.0 (MH*). 66b. (E)-N-(Piperidin-3-yI)-3-(2,6,6-trimethylcyclohex-1 -en-1 -yI)acrylamide hydrochloride The title compound, obtained as a pale brown amorphous solid (85 mg, 10 99%), by following the procedure of Example 49b. Rf = 0.1 (90:10) dichloromethane: methanol + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDCl 3 ) 8 9.61-9.53 (m,2H), 8.03 (s, 1H), 7.33 (d, J = 14.5 Hz, 1H), 6.00 (d, J = 14.5 Hz, 1H), 4.51 (s, 1H), 3.30-3.01 (m, 4H), 2.01-1.98 (m, 1H), 1.74 (s, 2H), 1.58 (s, 3H), 1.45-1.43 (m, 2H), 1.24-1.22 (m, 2H), 1.04 (s, 6H); Mass 15 spectrum (ESI +ve) m/z 277 (MH*). Example 67: (E)-N-Methyl-N-(piperidin-3-yl)-3-(2,6,6-trimethylcyclohex-1 en-1-yl)acrylamide hydrochloride 20 67a. (E)-tert-Butyl 4-(N-methyl-3-(2,6,6-trimethylcyclohex-1-en-1-yl) acrylamido)piperidine-1-carboxylate The title compound, obtained as a yellow oil (195 mg, 49%), was prepared from the product of Example la by following the procedure of Example 3 except tert-butyl 4-(methylamino)piperidine-1-carboxylate was 25 substituted for methylamine hydrochloride. Rf = 0.25 in (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.35 (d, J = 14.4, 1H), 6.34 (d, J = 14.4 Hz, 1H), 4.71 (s, 1H), 4.21 (s, 2H), 2.89 (s, 3H), 2.87-2.66 (m, 2H), 2.04 2.02 (m, 2H), 1.75 (s, 3H), 1.65-1.58 (m, 6H), 1.48-1.46 (m, 11H), 1.05 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 391.0 (MH*).

139 67b. (E)-N-Methy -N-(piperidin-3-yl)-3-(2,6,6-trimethylcyclohex-1-en-1 yl)acrylamide hydrochloride The title compound, obtained as a yellow film (160 mg, 99%), was prepared by following the procedure of Example 49b. Rf = 0.3 (90:10 5 dichloromethane: methanol + 0.1% (v/v) ammonium hydroxide); 1 H-NMR (400 MHz, CDC1 3 ) 5 7.30 (d, J = 15.0, 1H), 6.34 (d, J = 15.0 Hz, 1H), 4.66 (s, 1H), 3.52-3.49 (m, 2H), 3.17-3.11 (m, 2H), 3.03-2.94 (m, 3H), 2.10-2.07 (m, 4H), 1.91-1.87 (m, 2H), 1.78 (s, 3H), 1.67-1.64 (m, 2H), 1.52-1.49 (m, 2H), 1.07 (s, 6H), 0.89-0.85 (m, 2H); Mass spectrum (ESI +ve) m/z 291 (MH*). 10 Example 68: (E)-1-(1,1-Dioxidothiomorpholino)-3-(2,6,6 trimethylcyclohex-1 -eyl)prop-2-en-1 -one The title compound, obtained as a pale yellow waxy solid (28.2 mg, 15 18%), was prepared from the product of Example la by following the procedure of Example 3 except thiomorpholine 1,1-dioxide was substituted for methylamine hydrochloride. Rf = 0.10 (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCI 3 ) 5 7.42 (d, J = 15.5, 1H), 6.20 (d, J = 15.5 Hz, 1H), 4.10 4.07 (m, 4H), 3.10-2.98 (m, 4H), 2.07-1.98 (m, 2H), 1.75 (s, 3H), 1.61 (t, J = 20 6.0 Hz, 2H), 1.48 (t, J = 6.0 Hz, 2H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 312 (MH*). Example 69: (E)-1-Thiomorpholino-3-(2,6,6-trimethylcyclohex-1-en-1 yl)prop-2-en-1 -one 25 The title compound, obtained as a clear oil (104 mg, 73%), was prepared from the product of Example la by following the procedure of Example 3 except thiomorpholine was substituted for methylamine hydrochloride. Rf = 0.20 (10:90 ethyl acetate: hexanes); 'H-NMR (400 MHz, 30 CDCl 3 ) 5 7.32 (d, J = 15.5, 1H), 6.18 (d, J = 15.5 Hz, 1H), 3.95-3.85 (m, 4H), 140 2.67-2.65 (m, 4H), 2.03 (t, J = 6.0 Hz, 2H), 1.74 (s, 3H), 1.61 (t, J = 6.0 Hz, 2H), 1.46 (t, J = 6.0 Hz, 2H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 280 (MH*). 5 Example 70: (E)-1-(4,4-Difluoropiperidin-1-yl)-3-(2,6,6-trimethylcyclohex 1-en-1 -yl)prop-2-en-1 -one The title compound, obtained as a white solid (155 mg, 81%), was prepared by following the procedure of Example 3, except 4,4 difluoropiperidine was substituted for methylamine hydrochloride. Mp = 67.3 10 70.7*C; Rf = 0.44 (20:80 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 5 7.37 (d, J = 15.5 Hz, 1H), 6.24 (d, J = 15.5 Hz, 1H), 3.75 (br m, 4H), 2.09 1.96 (m, 6H), 1.76 (s, 3H), 1.63 (m, 2H), 1.55-1.45 (m, 2H), 1.07 (s, 6H); Mass spectrum (ESI +ve) m/z 298 (MH*). 15 Example 71: (±)-4-((E)-3-((1,6-anti)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1 -carboxamide 71a. (±)-(E)-Ethyl 3-((1,6-anti)-2,2,6-trimethylcyclohexyl)acrylate In a round bottom flask, sodium hydride (60% dispersion, 0.780 g, 19.4 20 mmol) was suspended in hexanes (10 mL) and the solvent was decanted. The residue was suspended in anhydrous tetrahydrofuran (40 mL) and the reaction flask was charged with argon and cooled to 0*C. To the stirred slurry was added triethylphosphonoacetate (3.24 mL, 3.63 g, 16.2 mmol) dropwise as to prevent build-up of the foaming reaction mixture. The reaction mixture 25 was stirred for 30 minutes while warming to room temperature until a clear solution remained. To this stirred solution was added a solution of (1,6-anti) 2,2,6-trimethylcyclohexanecarbaldehyde (2.00 g, 12.9 mmol) in anhydrous tetrahydrofuran (10 mL). The reaction was heated to reflux and stirred for 18 hours.

141 Upon cooling to room temperature, the reaction mixture was transferred to a separatory funnel and diluted with ethyl acetate (300 mL). The organic phase was extracted with a 50% saturated solution of ammonium chloride (2 x 100 mL) and then washed with brine (100 mL), dried over 5 magnesium sulfate, filtered and concentrated in vacuo to give a brown crude oil (-5 g). The title compound was purified by flash column chromatography (solvent gradient of 1-10% ethyl acetate: hexanes) to yield a pale brown oil (0.726 g, 28%). Rf = 0.50 (5:95 ethyl acetate: hexanes); 'H NMR (400 MHz,

CDC

3 ) 8 6.71 (dd, J = 15.5, 10.0 Hz, 1H), 5.74 (d, J = 15.5 Hz, 1H), 4.16 (q, J 10 = 7.0 Hz, 2H), 1.76-1.65 (m, 1H), 1.57-1.35 (m, 5H), 1.27 (t, J = 7.0 Hz, 3H), 1.15 (m, 1H), 0.93-0.88 (m, 1H), 0.86 (s, 3H), 0.80 (s, 3H), 0.72 (d, J = 6 Hz, 3H); Mass spectrum (ESI +ve) m/z 225 (MH*). 71b. (±)-(E)-3-((1,6-anti)-2,2,6-Trimethylcyclohexyl)acrylic acid (E)-Ethyl 3-((1,6-anti)-2,2,6-trimethylcyclohexyl)acrylate (71a, 1.03 g, 15 4.59 mmol) was dissolved in a 2:1 mixture of tetrahydrofuran (16 mL) and water (8 mL) and lithium hydroxide (0.549 g, 22.9 mmol) was added to the solution. The reaction mixture was heated to reflux and stirred overnight for 18 hours. The reaction mixture was transferred to a separatory funnel and diluted 20 with a 1M solution of sodium hydroxide (100 mL) and extracted with hexanes (50 mL). The aqueous phase was then acidified by adding concentrated hydrochloric acid (-16 mL) and then extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over magnesium sulfate, filtered and concentrated in vacuo to give a pale yellow 25 solid (0.61 g, 67%). Rf = 0.20 (25:75 ethyl acetate: hexanes); 'H NMR (400 MHz, CDCI 3 ) 8 10.50 (br s, 1H), 6.86 (dd, J = 15.5, 10.0 Hz, 1H), 5.78 (d, J = 15.5 Hz, 1H), 1.73 (m, 1H), 1.50 (m, 5H), 1.25-1.12 (m, 1H), 0.98-0.91 (m, 1H), 0.89 (s, 3H), 0.83 (s, 3H), 0.75 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 197 (MH*). 30 71c. (±)-tert-Butyl 4-((E)-3-((1,6-anti)-2,2,6-Trimethylcyclohexyl)acryloyl) piperazine-1-carboxylate 142 The title compound, obtained as a white solid (0.40 g, 95%), was prepared from the product of Example 71b by following the procedure of Example 3 except tert-butyl piperazine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.40 in (25:75 ethyl acetate: hexanes); 'H 5 NMR (400 MHz, CDCl 3 ) 8 6.69 (dd, J = 15.0, 10.0 Hz, 1H), 6.16 (d, J = 15.0 Hz, 1H), 3.73-3.39 (m, 8H), 1.74 (m, 1H), 1.60-1.39 (m, 15H), 1.22-1.12 (m, 1H), 0.90 (s, 3H), 0.83 (s, 3H), 0.77 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 365 (MH*). 71d. 4-((E)-3-((1,6-anti)-2,2,6-Trimethylcyclohexyl)acryloyl)piperazine-1 10 carboxamide To a solution of tert-butyl 4-((E)-3-((1,6-anti)-2,2,6 trimethylcyclohexyl)acryloyl)piperazine-1-carboxylate (71c, 0.35 g, 0.96 mmol) in dichloromethane (10 mL) was added dropwise a 4.0 M solution of hydrochloric acid in 1,4-dioxane (1.2 mL, 4.8 mmol). The reaction mixture was 15 stirred at room temperature for 18 hours then concentrated in vacuo. The crude oil was dissolved in dichloromethane (10 mL) and potassium carbonate (0.67 g, 4.8 mmol) and trimethylsilyl isocyanate (1.3 mL, 9.6 mmol) were added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 18 hours. 20 The reaction mixture was poured into a saturated aqueous solution of ammonium chloride (10 mL) and was extracted with dichloromethane (3 x 10 mL). The combined organic phases were dried over sodium sulfate and the concentrated in vacuo. The product was purified by flash column chromatography to yield the title compound as a white solid (0.205 mg, 69%). 25 Rf = 0.20 in (5:95 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 6.68 (dd, J = 15.0, 10.0 Hz, 1H), 6.13 (d, J = 15.0 Hz, 1H), 4.76 (br s, 2H), 3.52 (m, 8H), 1.91 (s, 1H), 1.72 (m, 1H), 1.50 (m, 5H), 1.15 (m, 1H), 0.87 (s, 3H), 0.81 (s, 3H), 0.74 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 308 (MH*). 30 143 Example 72: (-)-4-((E)-3-((1R, 6R)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxamide 72a. (-)-(E)-Ethyl 3-((1R,6R)-2,2,6-trimethylcy clohexyl)acrylate 5 The title compound, obtained as a pale brown oil (1.3 g, 18%), was prepared by following the procedure of Example 71a except (1R,6R)-2,2,6 trimethylcyclohexanecarbaldehyde was substituted for (1,6-anti)-2,2,6 trimethylcyclohexanecarbaldehyde. [a]D 2 3 = -15.3' (c = 0.25, CHCl 3 ); Rf= 0.50 (5:95 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 8 6.71 (dd, J = 10 15.5, 10.0 Hz, 1H), 5.74 (d, J = 15.5 Hz, 1H), 4.16 (q, J = 7.0 Hz, 2H), 1.76 1.65 (m, 1H), 1.57-1.35 (m, 5H), 1.27 (t, J = 7.0 Hz, 3H), 1.15 (m, 1H), 0.93 0.88 (m, 1H), 0.86 (s, 3H), 0.80 (s, 3H), 0.72 (d, J = 6 Hz, 3H); Mass spectrum (ESI +ve) m/z 225 (MH*). 72b. (-)-(E)-3-((1R,6R)-2,2,6-Trimethylcyclohexyl)acrylic acid 15 The title compound, obtained as a pale yellow solid (1.02 g, 91%), was prepared from the product of 72a by following the procedure of Example 60c. [a]D23 = o20.0' (c = 0.25, CHCl 3 ); Rf= 0.20 (25:75 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 8 10.50 (br s, 1H), 6.86 (dd, J = 15.5, 10.0 Hz, 1H), 5.78 (d, J = 15.5 Hz, 1H), 1.73 (m, 1H), 1.50 (m, 5H), 1.25-1.12 (m, 1H), 0.98 20 0.91 (m, 1H), 0.89 (s, 3H), 0.83 (s, 3H), 0.75 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 197 (MH*). 72c. (-)-tert-Butyl 4-((E)-3-((1R,6R)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1 -carboxylate The title compound, obtained as a white solid (0.16 g, 86%), was 25 prepared from the product of Example 72b by following the procedure of Example 3 except tert-butyl piperazine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.40 in (25:75 ethyl acetate: hexanes); 1

H

NMR (400 MHz, CDCl 3 ) 8 6.69 (dd, J = 15.0, 10.0 Hz, 1H), 6.16 (d, J = 15.0 Hz, 1H), 3.73-3.39 (m, 8H), 1.74 (m, 1H), 1.60-1.39 (m, 15H), 1.22-1.12 (m, 30 1H), 0.90 (s, 3H), 0.83 (s, 3H), 0.77 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 365 (MH*).

144 72d. (-)-4-((E)-3-((1R,6R)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 carboxamide The title compound, obtained as a white solid (85 mg, 63%), was prepared from the product of example 72c by following the procedure of 5 Example 71d. []D123 = g22.4o (c = 0.25, CHCl 3 ); 1 H-NMR (400 MHz, CDCl 3 ) 6 6.68 (dd, J = 15.0, 10.0 Hz, 1H), 6.13 (d, J = 15.0 Hz, 1H), 4.76 (br s, 2H), 3.52 (m, 8H), 1.91 (s, 1H), 1.72 (m, 1H), 1.50 (m, 5H), 1.15 (m, 1H), 0.87 (s, 3H), 0.81 (s, 3H), 0.74 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 308 (MH*). 10 Example 73: (+)-4-((E)-3-((1S,6S)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1 -carboxamide 73a. (+)-(E)-Ethyl 3-((1S,6S)-2,2,6-trimethylcyclohexyl)acry late 15 The title compound, obtained as a pale brown oil (1.4 g, 28%), was prepared by following the procedure of Example 71a except (1S,6S)-2,2,6 trimethylcyclohexanecarbaldehyde was substituted for (1,6-anti)-2,2,6 trimethylcyclohexanecarbaldehyde. [a]D23 = +16.8* (c = 0.25, CHCI 3 ); Rf = 0.50 (5:95 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDCl 3 ) 5 6.71 (dd, J = 20 15.5, 10.0 Hz, 1H), 5.74 (d, J = 15.5 Hz, 1H), 4.16 (q, J = 7.0 Hz, 2H), 1.76 1.65 (m, 1H), 1.57-1.35 (m, 5H), 1.27 (t, J = 7.0 Hz, 3H), 1.15 (m, 1H), 0.93 0.88 (m, 1H), 0.86 (s, 3H), 0.80 (s, 3H), 0.72 (d, J = 6 Hz, 3H); Mass spectrum (ESI +ve) m/z 225 (MH*). 73b. (+)-(E)-3-((1S,6S)-2,2,6-Trimethylcyclohexyl)acrylic acid 25 The title compound, obtained as a pale yellow solid (0.61 g, 67%), was prepared from the product of 73a by following the procedure of Example 60c. [a]D23 = +21.6* (c = 0.25, CHCl 3 ); Rf= 0.20 (25:75 ethyl acetate: hexanes); 1 H NMR (400 MHz, CDC13) 5 10.50 (br s, 1H), 6.86 (dd, J = 15.5, 10.0 Hz, 1H), 5.78 (d, J = 15.5 Hz, 1H), 1.73 (m, 1H), 1.50 (m, 5H), 1.25-1.12 (m, 1H), 0.98- 145 0.91 (m, 1H), 0.89 (s, 3H), 0.83 (s, 3H), 0.75 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 197 (MH*). 73c. (+)-tert-Butyl 4-((E)-3-((1S,6S)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1 -carboxylate 5 The title compound, obtained as a white solid (0.40 g, 98%), was prepared from the product of Example 73b by following the procedure of Example 3 except tert-butyl piperazine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.40 in (25:75 ethyl acetate: hexanes); 1

H

NMR (400 MHz, CDCl 3 ) 8 6.69 (dd, J = 15.0, 10.0 Hz, 1H), 6.16 (d, J = 15.0 10 Hz, 1H), 3.73-3.39 (m, 8H), 1.74 (m, 1H), 1.60-1.39 (m, 15H), 1.22-1.12 (m, 1H), 0.90 (s, 3H), 0.83 (s, 3H), 0.77 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 365 (MH*). 73d. (+)-4-((E)-3-((1S,6S)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 carboxamide 15 The title compound, obtained as a white solid (0.21 g, 69%), was prepared from the product of example 73c by following the procedure of Example 71d. [a]D 23 = +19.6* (c = 0.25, CHCl 3 ); 1 H-NMR (400 MHz, CDCl 3 ) 6 6.68 (dd, J = 15.0, 10.0 Hz, 1H), 6.13 (d, J =-15.0 Hz, 1H), 4.76 (br s, 2H), 3.52 (m, 8H), 1.91 (s, 1H), 1.72 (m, 1H), 1.50 (m, 5H), 1.15 (m, 1H), 0.87 (s, 20 3H), 0.81 (s, 3H), 0.74 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 308 (MH*). Example 74: (E)-1-Morpholino-3-((1R,6R)-2,2,6-trimethylcyclohexyl)prop 2-en-1-one 25 The title compound, obtained as a clear viscous oil (57.0 mg, 86%), was prepared from the product of Example 72b by following the procedure of Example 3 except morpholine was substituted for methylamine hydrochloride. [a]D 23 = -21.6o (c = 0.32, chloroform);' Rf = 0.30 in (10:90 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDC 3 ) 8 6.81 (dd, J = 15.0, 8.5 Hz, 1H), 6.12 30 (d, J = 15.0 Hz, 1H), 3.69-3.56 (m, 8H), 1.75-1.71 (m, 2H), 1.51-1.35 (m, 5H), 146 0.88-0.87 (m, 4H), 0.82 (s, 3H), 0.75 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 266 (MH*). Example 75: (E)-1-Thiomorpholino-3-((1R,6R)-2,2,6-trimethylcyclohexyl) 5 prop-2-en-1-one The title compound, obtained as a clear oil (38.0 mg, 54%), was prepared from the product of Example 72b by following the procedure of Example 3 except thiomorpholine was substituted for methylamine hydrochloride. [a123 = -20.40 (c = 0.32, chloroform); Rf = 0.40 in (10:90 ethyl 10 acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 6 6.63 (dd, J = 15.0, 8.5 Hz, 1H), 6.18 (d, J = 15.0 Hz, 1H), 3.90-3.82 (m, 4H), 2.62 (s, 4H), 1.73-1.70 (m, 1H), 1.49-1.47 (m, 5H), 1.13-1.12 (m, 1H), 0.87-0.84 (m, 4H), 0.89 (s, 3H), 0.74 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 282.1 (MH*). 15 Example 76: (E)-4-(3-(2,5,6,6-tetramethylcyclohex-2-en-1-yl)acryloyl) piperazine-1 -carboxamide 76a. (E)-3-(2,5,6,6-tetramethylcyclohex-2-en-1-yl)acrylic acid The title compound, obtained as a clear viscous oil (1.90 g, 50%), was 20 prepared from irone (290% a:p isomers) by following the procedure of Example 1. Rf = 0.6 (25:75 ethyl acetate: hexanes + 0.1% (v/v) acetic acid); 1 H-NMR (400 MHz, CDCl 3 ) 6 11.71 (br s, 1H), 7.02-6.91 (m, 1H), 5.83 (q, J = 15.0 Hz, 1H), 5.49 (d, J = 17.5 Hz, 1H), 2.57 (d, J = 11.0 Hz, 0.43 H), 2.29 (d, J = 9.5 Hz, 0.57H), 2.03-1.91 (m, 1H), 1.69-1.67 (m, 2H), 1.54 (d, J = 16.5 Hz, 25 1H) 1.10 (br s, 0.43H), 0.87-0.81 (m, 8H), 0.70 (s, 1H); Mass spectrum (ESI ve) m/z 207 (MH~). 76b. (E)-4-(3-(2,5,6,6-tetramethylcyclohex-2-en-1-yl)acryloyl)piperazine-1 carboxamide To a solution of (E)-3-(2,5,6,6-tetramethylcyclohex-2-en-1-yl)acrylic 30 acid (76a, 100 mg, 0.480 mmol) in acetonitrile (5.0 mL) was added 2-(7-aza- 147 1 H-benzotriazole-1 -yl)-1, 1,3,3-tetramethyluronium hexafluorophosphate (HATU, 182 mg, 0.480 mmol). The solution was stirred at room temperature for 30 minutes then diisopropylethylamine (62.0 mg, 0.480 mmol) and tert butyl piperazine-1-carboxylate (89.4 mg, 0.480 mmol) was added to the 5 reaction mixture. The reaction was then stirred at 40'C for 4 hours. The reaction was quenched with a 1M solution of hydrochloric acid (2 mL) and the biphasic mixture was separated. The organic layer was concentrated in vacuo (400C) and the crude material loaded on to silica gel for purification via flash column chromatography running an isocratic eluent of 10 30% ethyl acetate in hexanes. The intermediate was isolated as a white solid (120 mg, 66%). The intermediate was dissolved in dichloromethane (10 mL) was added dropwise a 4.0 M solution of hydrochloric acid in 1,4-dioxane (1.2 mL, 4.8 mmol). The reaction mixture was stirred at room temperature for 18 hours 15 then concentrated in vacuo. The crude oil was dissolved in dichloromethane (10 mL) and potassium carbonate (0.67 g, 4.8 mmol) and trimethylsilyl isocyanate (1.3 mL, 9.6 mmol) were added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 18 hours. 20 The reaction mixture was poured into a saturated aqueous solution of ammonium chloride (10 mL) and was extracted with dichloromethane (3 x 10 mL). The combined organic phases were dried over sodium sulfate and the concentrated in vacuo. The product was purified by preparative plate thin layer chromatography to yield the title compound as a clear oil (13.0 mg, 25 11%). Rf = 0.40 (5:95 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 6.85-6.74 (m, 1H), 6.14 (m, 1H), 5.46 (d, J = 18.0 Hz, 1H), 4.85 (s, 2H), 3.72 3.41 (m, 8H), 2.26 (d, J = 11.0 Hz, 0.44H), 2.03 (d, J = 9.5 Hz, 0.57H), 2.04 1.90 (m, 1H), 1.69-1.64 (m, 1H), 1.54 (d, J = 15.0 Hz, 3H), 0.87-0.80 (m, 8H), 0.71 (s, 1H); Mass spectrum (ESI +ve) m/z 320 (MH*). 30 148 Example 77: 4-((E)-3-((1R,6S)-2,2,6 trimethylcyclohexyl)acryloyl)piperazine-l-carboxamide and 4-((E)-3 ((1S,6R)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxamide 5 77a. (±)-1,6-syn-3-(2,2,6-trimethylcyclohexyl)propanoic acid The title compound, obtained as a clear viscous oil (316 mg, 16%), was prepared from tetrahydroionone (2.00 g, 10.1 mmol) by following the procedure of Example 1. Rf = 0.30 (10:90 ethyl acetate: hexanes + 0.1% (v/v) acetic acid); 1 H-NMR (400 MHz, CDCl 3 ) 6 10.19 (br s, 1H), 2.33 (t, J = 8.5 Hz, 10 2H), 1.93-1.90 (m, 1H), 1.62-1.58 (m, 2H), 1.46-1.44 (m, 3H), 1.33-1.29 (m, 2H), 1.11-1.09 (m, 2H), 0.95 (s, 3H), 0.94 (s, 6H); Mass spectrum (ESI -ve) m/z 197 (MH-). 77b. 4-((E)-3-((1 R,6S)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 carboxamide and 4-((E)-3-((1S,6R)-2,2,6-trimethylcyclohexyl)acryloyl) 15 piperazine-1 -carboxamide The title compound, obtained as a white film (17.0 mg, 36%), was prepared from the product of Example 77a by following the procedure of Example 76b. Rf = 0.50 (5:95 methanol: chloroform); 1 H-NMR (400 MHz, CDCla) 6 4.61 (s, 2H), 3.66 (t, J = 5.0 Hz, 2H), 3.49 (s, 4H), 3.36 (t, J = 5.0 Hz, 20 2H), 2.29 (t, J = 8.5 Hz, 2H), 1.93-1.89 (m, 1H), 1.32-1.29 (m, 5H), 1.16-1.43 (m, 3H), 1.10-1.02 (m, 3H), 0.95 (s, 3H), 0.89 (S, 3H), 0.87 (s, 3H) ppm; Mass spectrum (ESI +ve) m/z 310 (MH*). Example 78: (E)-4-(3-(2,6,6-trimethylcyclohex-2-en-1 25 yl)acryloyl)piperazine-1-carboxamide 78a. (E)-3-(2,6,6-trimethylcyclohex-2-en-1-yl)acrylic acid The title compound, obtained as a clear viscous oil (56 mg, 3%), was prepared from x-ionone (2.00 g, 10.3 mmol) by following the procedure of 30 Example 1. Rf = 0.2 (10:90 ethyl acetate: hexanes + 0.1% (v/v) acetic acid); 1 H-NMR (400 MHz, CDCl) 6 11.33 (br s, 1H), 6.95-6.88 (m, 1H), 5.81 (d, J = 149 15.0 Hz, 1H), 5.49 (s, 1H), 2.31-2.29 (m, 1H), 2.04 (s, 2H), 1.50 (s, 3H), 1.48 1.43 (m, 1H), 1.22-1.18 (m, 1H), 0.92 (s, 3H), 0.89 (s, 3H); Mass spectrum (ESI -ve) m/z 193 (MH-). 78b. (E)-4-(3-(2,6,6-trimethylcyclohex-2-en-1-yl)acryloyl)piperazine-1 5 carboxamide The title compound, obtained as a white solid (22.0 mg, 36%), was prepared from the product of Example 78a by following the procedure of Example 76b. Rf = 0.50 (5:95 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 6.75 (dd, J = 14.5, 9.5 Hz, 1H), 6.15 (d, J = 14.5 Hz, 1H), 5.47 (s, 10 1H), 4.72 (s, 2H), 3.59-3.40 (m, 8H), 2.27 (d, J = 9.5 Hz, 1H), 2.01 (br s, 2H), 1.56 (s, 3H), 1.50-1.42 (m, 1H), 1.24-1.18 (m, 1H), 0.97 (s, 3H), 0.84 (s, 3H); Mass spectrum (ESI +ve) m/z 306 (MH*). Example 79: (±)-4-((E)-3-(1,3,3-Trimethyl-7-oxabicyclo[4.1.0]heptan-2 15 yl)acryloyl) piperazine-1-carboxamide 79a. (±)-(E)-3-(1,3,3-trimethyl-7-oxabicyclo[4.1.0]heptan-2-yl)acrylic acid The title compound, obtained as a white solid (616 mg, 31%), was prepared from 4-(1,3,3-trimethyl-7-oxabicyclo[4.1.0]hept-2-yl)-3-buten-2-one 20 (290% cis:trans isomers) (2.00 g, 9.60 mmol) by following the procedure of Example 1. Mp = 122.2-130.6*C; Rf = 0.20 (10:90 ethyl acetate: hexanes + 0.1% (v/v) acetic acid); 1 H-NMR (400 MHz, CDCl 3 ) 8 11.84 (br s, 1H), 7.07 6.92 (m, 1H), 5.91 (d, J = 15.5 Hz, 1H), 3.07 (s, 1H), 2.10 (d, J = 11.0 Hz, 1H), 1.97-1.94 (m, 1H), 1.86-1.73 (m, 1H), 1.49-1.39 (m, 1H), 1.39 (s, 3H), 1.05 25 0.98 (m, 1H) 0.93 (s, 3H), 0.77 (s, 3H) ; Mass spectrum (ESI -ve) m/z 209 (MH-). 79b. (±)-4-((E)-3-(1,3,3-Trimethyl-7-oxabicyclo[4.1.0]heptan-2-yl)acryloyl) piperazine-1 -carboxamide The title compound, obtained as an off-white solid (19.9 mg, 44%), was 30 prepared from the product of Example 79a by following the procedure of 150 Example 76b. Rf = 0.30 (5:95 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 6.75 (dd, J = 15.0, 10.0 Hz, 1H), 6.27 (d, J = 15.0 Hz, 1H), 4.71 (s, 2H), 3.70-3.43 (m, 8H), 3.05 (s, 1H), 2.08 (d, J = 10.0 Hz, 1H), 1.96-87 (m, 2H), 1.43-1.41 (m, 1H), 1.25 (s, 3H), 0.97-0.90(m, 1H), 0.91 (s, 3H), 0.86 (s, 5 3H); Mass spectrum (ESI +ve) m/z 322 (MH*). Example 80: 4-(3-((1R,6S)-2,2,6 trimethylcyclohexyl)propanoyl)piperazine-1 -carboxamide 10 4-((E)-3-((1 S,6S)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 carboxamide (73d, 50.0 mg, 0.16 mmol) was dissolved in anhydrous methanol (5.0 mL) at room temperature under argon to which 10% palladium on carbon (3.00 mg, 5.00 mmol) was added and allowed to stir vigorously. The reaction vessel was evacuated under vacuum and then charged with 15 hydrogen gas, the reaction mixture was left to stir at room temperature for 2 days. The reaction mixture was filtered through Celite and the solvent was removed in vacuo to yield a white solid (50.0 mg, 99%). 1 H-NMR (400 MHz, CDCl 3 ) 8 4.67 (br s, 2H), 3.65-3.63 (m, 2H), 3.48 (br s, 4H), 3.67-3.45 (m, 2H), 20 2.40-2.26 (m, 2H), 1.75-1.60 (m, 2H), 1.44-1.33 (m, 5H), 1.17-1.09 (m, 1H), 0.93-0.87 (m, 7H), 0.80 (s, 3H), 0.59-0.57 (m, 1H); Mass spectrum (ESI +ve) m/z 310 (MH*). Example 81: 4-(3-((1S,6R)-2,2,6 25 trimethylcyclohexyl)propanoyl)piperazine-1 -carboxamide 4-((E)-3-((1 R,6R)-2,2,6-trimethylcyclohexyl)acryloyl)piperazine-1 carboxamide (72d, 28.0 mg, 0.09 mmol) was dissolved in anhydrous methanol (5.00 mL) at room temperature under argon to which 10% palladium 30 on carbon (3.00 mg, 5.00 mmol) was added and allowed to stir vigorously.

151 The reaction vessel was evacuated under vacuum and then charged with hydrogen gas, the reaction mixture was left to stir at room temperature for 2 days. The reaction mixture was filtered through Celite and the solvent was 5 removed in vacuo to yield a white solid (28.0 mg, 99%). 1 H-NMR (400 MHz, CDCl 3 ) 5 4.67 (br s, 2H), 3.65-3.63 (m, 2H), 3.48 (br s, 4H), 3.67-3.45 (m, 2H), 2.40-2.26 (m, 2H), 1.75-1.60 (m, 2H), 1.44-1.33 (m, 5H), 1.17-1.09 (m, 1H), 0.93-0.87 (m, 7H), 0.80 (s, 3H), 0.59-0.57 (m, 1H); Mass spectrum (ESI +ve) m/z 310 (MH*). 10 Example 82: (E)-4-(3-(2-chloro-3-hydroxy-2,6,6 trimethylcyclohexyl)acryloyl) piperazine-1-carboxamide To a solution of (±)-(E)-3-(1,3,3-trimethyl-7-oxabicyclo[4.1.0]heptan-2 15 yl)acrylic acid (79a, 100 mg, 0.476 mmol) in acetonitrile (5.0 mL) was added 2-(7-aza-1 H-benzotriazole-1 -yl)-1, 1,3,3-tetramethyluronium hexafluorophosphate (HATU, 181 mg, 0.476 mmol). The solution was stirred at room temperature for 30 minutes then diisopropylethylamine (61.5 mg, 0.476 mmol) and tert-butyl piperazine-1-carboxylate (88.6 mg, 0.476 mmol) 20 was added to the reaction mixture. The reaction was then stirred at 40 0 C for 4 hours. The reaction was quenched with a 1M solution of hydrochloric acid (2 mL) and the biphasic mixture was separated. The organic layer was concentrated in vacuo (40 0 C) and the crude material loaded on to silica gel for 25 purification via flash column chromatography running an isocratic eluent of 30% ethyl acetate in hexanes. The intermediate was isolated as a white solid (135 mg, 75%). The intermediate was dissolved in dichloromethane (10 mL) was added dropwise a 4.0 M solution of hydrochloric acid in 1,4-dioxane (1.2 mL, 4.8 30 mmol). The reaction mixture was stirred at room temperature for 18 hours then concentrated in vacuo.

152 The crude oil was dissolved in dichloromethane (10 mL) and potassium carbonate (0.67 g, 4.8 mmol) and trimethylsilyl isocyanate (1.3 mL, 9.6 mmol) were added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 18 hours. 5 The reaction mixture was poured into a saturated aqueous solution of ammonium chloride (10 mL) and was extracted with dichloromethane (3 x 10 mL). The combined organic phases were dried over sodium sulfate and the concentrated in vacuo. The product was purified by preparative plate thin layer chromatography to yield the title compound as a white solid (19.0 mg, 10 15%). Rf = 0.30 (10:90 methanol: chloroform); 1 H-NMR (400 MHz, CD 3 0D) 5 6.97 (dd, J = 15.5, 11.0 Hz, 1H), 6.46 (d, J = 15.5 Hz, 1H), 3.99 (s, 1H), 3.68 3.66 (m, 4H), 3.44-3.42 (m, 4H), 2.57-2.54 (m, 1H), 2.29-2.26 (m, 1H), 1.89 1.82 (m, 1H), 1.70-1.66 (m, 1H), 1.29-1.26 (m, 1H), 1.23-1.20 (m, 1H), 1.13 (s, 3H), 1.04 (s, 3H), 0.81 (s, 3H); Mass spectrum (ESI +ve) m/z 358 [ 35 C1, 15 35 C1], 359.0 [ 35 C1, 3 7 C1], 360.1 [ 37 C1, 37 C1] (MH*). Example 83: (E)-1-Morpholino-3-((1S,6S)-2,2,6-trimethylcyclohexyl)prop 2-en-1 -one 20 The title compound, obtained as a clear oil (62.1 mg, 74%), was prepared from the product of Example 71b by following the procedure of Example 3 except morpholine was substituted for methylamine hydrochloride. [a]D23 = +22.8* (c = 0.25, chloroform); Rf = 0.50 in (40:60 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 8 6.81 (dd, J = 15.0, 8.5 Hz, 1H), 6.12 25 (d, J = 15.0 Hz, 1H), 3.69-3.56 (m, 8H), 1.75-1.71 (m, 2H), 1.51-1.35 (m, 5H), 0.88-0.87 (m, 4H), 0.82 (s, 3H), 0.75 (d, J = 6.0 Hz, 3H); Mass spectrum (ESI +ve) m/z 266 (MH*). 30 153 Example 84: (E)-4-(3-(2,6,6-Trimethylcyclohex-1-en-1 yl)acryloyl)piperazin-2-one The title compound, obtained as a clear oil (165 mg, 60%), was prepared from the product of Example 1 by following the procedure of 5 Example 3 except piperazin-2-one was substituted for methylamine hydrochloride. Rf = 0.36 in (15:85 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.42 (d, J = 15.0 Hz, 1H), 6.98 (d, J = 2.0 Hz, 1H), 6.15 (d, J = 15.0 Hz, 1H), 4.23 (s, 2H), 3.83 (m, 2H), 3.43 (m, 2H), 2.03 (m, 2H), 1.84-1.37 (m, 7H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 277 (MH*). 10 Example 85: (E)-1-(1,4-Dioxa-8-azaspiro[4.5]decan-8-yI)-3-(2,6,6 trimethylcyclohex-1-en-1-yi) prop-2-en-1-one The title compound, obtained as a clear oil (295 mg, 92%), was prepared from the product of Example 1 by following the procedure of 15 Example 3 except 1,4-dioxa-8-azaspiro[4.5]decane was substituted for methylamine hydrochloride. Rf = 0.2 in (40:60 ethyl acetate: hexanes); 1

H

NMR (400 MHz, CDCl 3 ) 5 7.30 (d, J = 15.0 Hz, 1H), 6.23 (d, J = 15.0 Hz, 1H), 3.98 (s, 4H), 3.81-3.53 (m, 4H), 2.03 (t, J = 6.0 Hz, 1H), 1.73 (s, 3H), 1.72 1.53 (m, 2H), 1.49-1.44 (m, 2H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 20 320 (MH*). Example 86: (E)-Ethyl 1-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acryloyl) piperidi ne-4-carboxylate The title compound, obtained as a clear oil (286 mg, 83%), was 25 prepared from the product of Example 1 by following the procedure of Example 3 except ethyl piperidine-4-carboxylate was substituted for methylamine hydrochloride. Rf = 0.5 in (50:50 ethyl acetate: hexanes); 1

H

NMR (400 MHz, CDCl 3 ) 6 7.28 (d, J = 15.0 Hz, 1H), 6.19 (d, J = 15.0 Hz, 1H), 4.45 (br s, 1H), 4.13 (q, J = 7.0 Hz, 2H), 3.94 (br s, 1H), 3.01 (m, 2H), 2.53 (tt, 30 J = 10.5, 4.0 Hz, 1H), 2.15-1.84 (m, 4H), 1.72 (s, 3H), 1.63 (m, 4H), 1.45 (m, 154 2H), 1.24 (t, J = 7.0 Hz, 3H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 334 (MH*). Example 87: (E)-4-(3-(2,6,6-Trimethylcyclohex-1-en-1 5 yl)acryloyl)piperazine-1 -sulfonamide The title compound, obtained as a white solid (41.0 mg, 58%), was prepared from the product of Example 1 by following the procedure of Example 3 except piperazine-1-sulfonamide was substituted for methylamine hydrochloride. Rf = 0.6 in (10:90 methanol: chloroform); Mp = 181-183*C; 'H 10 NMR (400 MHz, CDC 3 ) 8 7.44-7.32 (d, J = 15.0 Hz, 1H), 6.25-6.14 (d, J = 15.0 Hz, 1H), 4.46 (br s, 2H), 3.75 (m, 4H), 3.21 (m, 4H), 2.04 (t, J = 6.05 H, 2H), 1.75 (s, 3H), 1.62 (m, 2H), 1.48 (m, 2H), 1.05 (s, 6H); Mass spectrum (ESI +ve) m/z 342 (MH*). 15 Example 88: (E)-4-(3-(2,6,6-Trimethylcyclohex-1-en-1 yl)acryloyl)piperazine-1 -carbaldehyde Acetic anhydride (0.25 mL) was added to a solution of formic acid (1.25 mL) at room temperature. To this stirred solution was added the product of Example 28b predissolved in formic acid (0.5 mL). The reaction was stirred at 20 room temperature for 16 hours. The reaction mixture was then concentrated into silica gel and purified by flash column chromatography to yield the title compound as a yellow oil (92.0 mg, 46%). Rf = 0.50 in (10:90 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 8.09 (s, 1H), 7.37 (d, J = 15.5 Hz, 1H), 6.19 (d, J = 15.5 Hz, 25 1H), 3.83-3.49 (m, 6H), 3.47-3.33 (m, 2H), 2.03 (t, J = 6.0 Hz, 2H), 1.73 (s, 3H), 1.60 (m, 2H), 1.52-1.36 (m, 2H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 291 (MH*).

155 Example 89: (E)-1-(4-(2-hydroxyethyl)piperazin-1-yI)-3- (2,6,6 trimethylcyclohex-1 -en-1 -yl)prop-2-en-1 -one The title compound, obtained as a clear oil (25.0 mg, 12%), was prepared from the product of example 28b by following the procedure of 5 Example 7c except 2-bromoethanol was substituted for iodomethane. Rf = 0.23 in (10:90 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.32 (d, J = 15.5 Hz, 1H), 6.19 (d, J = 15.5 Hz, 1H), 3.81-3.44 (m, 6H), 2.72 (s, 1H), 2.62-2.44 (m, 6H), 2.02 (t, J = 6.0 Hz, 2H), 1.73 (s, 3H), 1.67-1.53 (m, 2H), 1.50-1.37 (m, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 307 (MH*). 10 Example 90: (±)-3,5-cis-Dimethyl-4-((E)-3-(2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl) piperazine-1-carboxamide The title compound, obtained as a yellow amorphous solid (242 mg, 86%), was prepared from the product of Example 1 by following the procedure 15 of Example 76b except tert-butyl cis-3,5-dimethylpiperazine-1-carboxylate was substituted for tert-butyl piperazine-1-carboxylate. Rf = 0.10 in (10:90 methanol: chloroform); Mp = 152-155 0 C; 1 H-NMR (400 MHz, CDC 3 ) 6 7.38 (d, J = 15.5 Hz, 1H), 6.19 (d, J = 15.5 Hz, 1H), 4.70 (s, 2H), 3.78 (m, 4H), 3.15 (m Hz, 2H), 2.04 (t, J = 6.0 Hz, 2H), 1.74 (s, 3H), 1.61 (m, 2H), 1.52-1.41 (m, 2H), 20 1.33 (s, 3H), 1.32 (s, 3H), 1.09 (s, 6H); Mass spectrum (ESI +ve) m/z 334 (MH*). Example 91: (E)-4-(3-(2,2,6-trimethylbicyclo[4.1.0]heptan-1-yl)acryloyl) piperazine-1 -carboxamide 25 Sodium borohydride (6.50 mg, 172 mmol) was added to a stirred solution of @-cyclocitral (13.2 g, 86.0 mmol) in methanol (400 mL) at 0*C under argon. The reaction was stirred at room temperature for 6 hours. The reaction was quenched by adding water (400 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine 30 (600 mL), dried over magnesium sulfate, filtered and concentrated afford the alcohol as a clear oil (13.5 g, quantitative).

156 In a dry round bottom flask diethyl zinc (1.0M in hexanes, 8.60 mL, 8.60 mmol) was added to anhydrous diethyl ether (10 mL) at room temperature. To this solution was added dropwise methyleneiodide (0.71 mL, 8.85 mmol) and the reaction was stirred at room temperature for 15 minutes 5 resulting in the formation of a white precipitate. The above prepared alcohol (910 mg, 5.90 mmol) was dissolved in diethyl ether (4.0 mL) and added to the reaction mixture. The reaction was stirred a room temperature for 20 minutes and then heated to reflux for 16 hours. The reaction was cooled to OC and quenched with a saturated solution of ammonium chloride (2.0 mL). The 10 biphasic reaction mixture was transferred to a separatory funnel and diluted with diethyl ether (10 mL) and the extracted with saturated ammonium chloride (20 mL) and washed with brine (20 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated to give a brown oil (-1.1 g). The cyclopropanated alcohol was isolated by flash column chromatography 15 (0-20% ethyl acetate in hexanes) as a clear oil (385 mg, 39%). The cyclopropyl alcohol (371 mg, 2.20 mmol) was dissolved in dichloromethane (34 mL) and Dess-Martin periodinane (1.03 g, 2.43 mmol) was added to the stirred solution followed by a drop of water (0.05 mL). The reaction was stirred at room temperature for 1 hour then concentrated to 20 remove the dichloromethane. The residue was dissolved in diethyl ether (80 mL) and treated with a 1:1 (v/v) solution of 10% sodium thiosulfate (25 mL) and saturated sodium bicarbonate (25 mL) for 30 minutes. The layers were separated and the organic phase was washed with water (50 mL) and brine (50 mL), then dried over sodium sulfate, filtered and concentrated to give a 25 crude solid. The aldehyde was isolated by flash column chromatography (0 20% diethyl ether in hexanes) as a grey solid (260 mg, 71%). The aldehyde was carried forward according to the procedure described in Examples 71a through 71d to furnish the title compound as a white solid (35.1 mg, 94%). Rf = 0.17 in (5:95 methanol: chloroform); Mp = 30 166-168'C; 1 H-NMR (400 MHz, CD 3 0D) 6 7.14 (dd, J = 15.0, 3.0 Hz, 1H), 6.35 (dd, J = 15.0, 3.0 Hz, 1H), 3.66 (m, 4H), 3.46 (m, 4H), 1.81-1.66 (m, 2H), 157 1.62-1.25 (m, 3H), 1.25-1.10 (m, 6H), 0.99 (d, J = 3.0 Hz, 3H), 0.91 (t, J = 6.0 Hz, 3H), 0.67 (m, 2H); Mass spectrum (ESI +ve) m/z 320 (MH*). Example 92: (E)-4-(3-(4-hydroxy-2,6,6-trimethylcyclohex-1-enyl)acryloyl) 5 piperazine-1-carboxamide In 1 L Parr vessel reactor 4-oxo-isophorone (20.0 g, 130 mmol) was dissolved in ethanol (200 mL) and Raney/Ni (0.1 eq) was added to the solution. The vessel was charged with hydrogen gas to a pressure of 100 psi. The reaction mixture was stirred at room temperature for 3 days, filtered 10 through a pad of Celite, and the solvent was removed under reduced pressure to yield the product as clear oil, the crude was carried forward into the next step of synthesis. To a solution of crude 4-hydroxy-2,2,6- trimethylcyclohexan-1-one (20.2 g, 130 mmol) and imidazole (35.3 g, 520 mmol) in dichloromethane (200 15 mL) at 0*C, was added a solution of tert-butylchlorodimethylsilane (78.3 g, 260 mmol) in dichloromethane (200 mL). The reaction was stirred for 16 h and then poured into water (100 mL) and extracted with hexane (3 x 150 mL). The organic layer was washed with water (5 x 100 mL), dried over magnesium sulfate and concentrated. The residue was purified by flash column 20 chromatography (silica gel, 97:3 hexane:ethyl acetate) to afford 14.0 g (40%) of 4-(tert-butyldimethylsilyloxy)-2,2,6-trimethyl cyclohexan-1-one as colorless oil. To a solution of 4-(tert-butyldimethylsilyloxy)-2,2,6-trimethylcyclohexan 1-one (7.00 g, 26.0 mmol) in ethanol (50 mL) at 25 0 C, were added hydrazine 25 monohydrate (33.5 g, 670 mmol) and diisopropylethylamine (9.80 mL, 56.3 mmol). After the mixture was stirred for 24 h at 100 0 C, the solvent was removed and the residue was taken in diethyl ether (30 mL) and washed with brine (3 x 50 mL). The aqueous layers were extracted with diethyl ether (4 x 50 mL), and the organic extracts were dried over magnesium sulfate and 30 concentrated.

158 To a solution of the residue in diethyl ether (30 mL) and 1,5 diazabicyclo[4.3.0.] nonane (25.0 mL, 200 mmol) was added a solution of iodine (9.90 g, 39.0 mmol) in diethyl ether (30 mL). After the mixture was stirred for 15 min, an aqueous solution of saturated sodium bicarbonate was 5 added, the layers were separated, the organic layer was dried over sodium sulfate, and the solvent was removed. A solution of the residue in benzene (60 mL) was treated with 1,5-diazabicyclo[4.3.0.] nonane (25 mL). The mixture was stirred for 2.5 h, then poured into diethyl ether (200 mL) and washed with aqueous sodium thiosulfate (3 x 30 mL), and the organic layer 10 was dried and evaporated. The residue was purified by chromatography (silica gel, 5% ethyl acetate:hexanes) to afford 5.2 g (53%) of ter butyldimethylsilyl-3,5,5-trimethyl-4-iodocyclohex-3-en-1-yI ether. To a solution of the ether (0.80 g, 2.21 mmol) in N,N-dimethylforamide (10 mL) was added tetrakistriphenylphosphine palladium (0.240 g, 0.210 15 mmol), and the mixture was degassed by the freeze-thaw method (three cycles). Methyl vinyl ketone (0.530 mL, 6.31 mmol) and triethylamine (0.880 mL, 6.31 mmol) were then added, and the reaction was heated to 170 0 C for 1 h using microwave irradiation. The mixture was diluted with diethyl ether (50 mL), washed with a 1% solution of hydrochloric acid, and extracted with Et 2 0 20 (3 x 25 mL). The combined organic layers were washed with a saturated solution of aqueous sodium bicarbonate (3 x 25 mL) and dried over magnesium sulfate, and the solvent was removed. The resulting oil was purified by flash column chromatography (silica gel, 90:10 hexanes:ethyl acetate) affording 314 mg (42%) of [(E)-4-(tert-butyldimethylsilyloxy)-2,6,6 25 trimethylcyclohex-1-en-1-yl]but-3-en-2-one as colorless oil. The ketone was hydrolyzed to the carboxylic acid according to the procedure outlined in Example 1, and the corresponding acrylic acid was carried forward according to the procedure described in Examples 71b through 71d to furnish the title compound as a clear oil (10.1 mg, 30%). Rf = 30 0.20 in (10:90 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 8 6.19 (d, J = 15.5 Hz, 1H), 4.66 (s, 2H), 4.08-3.91 (m, 1H), 3.84-3.34 (m, 8H), 2.40 (dd, J = 17.0, 5.5 Hz, 1H), 2.14-1.98 (m, 1H), 1.87-1.65 (m, 5H), 1.47 (t, J = 12.0 Hz, 159 1H), 7.31 (d, J = 15.5 Hz, 1H), 1.09 (s, 3H), 1.08 (s, 3H); Mass spectrum (ESI +ve) m/z 322 (MH*). Example 93: (±)-(E)-4-(3-(4-Methoxy-2,6,6-trimethylcyclohex-1-en-1-yI) 5 acryloyl) piperazine-1-carboxamide To a solution of [4-(tert-butyIdimethylsilyloxy)-2,6,6-trimethylcyclohex-l en-1-yl]but-3-en-2 one (230 mg, 0.680 mmol), prepared in Example 92, in tetrahydrofuran (2.0 mL) was added tetrabutylammonium fluoride (1.0 M in THF, 2.00 mL, 2.00 mmol), and the mixture was stirred for 16 h at room 10 temperature. The reaction was poured on to an aqueous solution of saturated sodium bicarbonate, and extracted with diethyl ether (3 x 10 mL), and dried over magnesium sulfate, and the solvent was evaporated. The residue was purified by flash column chromatography (silica gel, 60:20 hexanes:ethyl acetate), affording 119 mg (78%) of the intermediate hydroxy ketone. 15 The hydroxy-ketone was hydrolyzed to the carboxylic acid according to the procedure outlined in Example 1. The corresponding hydroxy-acid (117 mg, 0.52 mmol) was dissolved in diethyl ether (5 mL) at 0*C, and a solution of diazomethane in diethyl ether (5 mL) was added followed by boron trifluoride diethyl etherate (3 drops). A white precipitate formed and nitrogen gas was 20 evolved. After 30 min, the mixture was filtered and the filtrate was concentrated. Purification of the residue by flash column chromatography on silica gel (70:30 hexanes:ethyl acetate) gave the 4-methoxy-methyl ester as a colorless oil (67 mg, 51%). The methyl ester was carried forward according to the procedure described in Examples 71b through 71d to furnish the title 25 compound as a white solid (27 mg, 30%). Rf = 0.34 in (10:90 methanol: chloroform); 1 H-NMR (400 MHz, CDCI 3 ) 5 7.32 (d, J = 15.5 Hz, 1H), 6.20 (d, J = 15.5 Hz, 1H), 4.63 (s, 2H), 3.82-3.40 (m, 9H), 3.38 (d, J = 11.50 Hz, 3H), 2.43 (dd, J = 17.5, 5.5 Hz, 1H), 2.03 (dd, J = 17.5, 9.5 Hz, 1H), 1.83 (d, J = 13.5 Hz, 1H), 1.76 (s, 3H), 1.40 (t, J = 12.0 Hz, 1H), 1.09 (s, 3H), 1.08 (s, 3H); 30 Mass spectrum (ESI +ve) m/z 336 (MH*).

160 Example 94: (-)-((1 R,6S)-2,2,6-trimethylcy clohexyl)methyl 4 carbamoylpiperazine-1 -carboxylate Sodium borohydride (78.0 mg, 2.07 mmol) was added to a stirred solution of (1S,6S)-2,2,6-trimethylcyclohexanecarbaldehyde (320 mg, 2.07 5 mmol) in methanol (10 mL) at 0*C under argon. The reaction was stirred at room temperature for 16 hours. The reaction was quenched by adding water (60 mL) and extracted with diethyl ether (4 x 25 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The intermediate alcohol was purified by flash column chromatography (0-25% 10 ethyl acetate in hexanes) and obtained as a clear oil (248 mg, 77%). The alcohol (235 mg, 1.51 mmol) was dissolved in anhydrous dichloromethane (20 mL) and cooled to 0*C. To this stirred solution was added triphosgene (179 mg, 0.61 mmol) and pyridine (227 mg, 2.88 mmol). The reaction was stirred at 0*C until complete consumption of the starting 15 alcohol by TLC. tert-Butyl piperazine-1-carboxylate (338 mg, 1.81 mmol) was then added to the reaction mixture in one portion. The reaction was stirred at room temperature for 18 hours. The reaction mixture was then diluted with dichloromethane (30 mL) and extracted with 1M HCI (2 x 15 mL) and saturated NaHCO 3 solution (15 mL). The organic layer was washed with brine 20 (20 mL) then dried over sodium sulfate, filtered and concentrated. The intermediate was purified by flash column chromatography (0-25% ethyl acetate in hexanes) and obtained as a clear oil (394 mg, 70%). The Boc-protected intermediate was carried forward following the procedure of Example 71d and was substituted for tert-butyl 4-((E)-3-((1,6 25 anti)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxylate (71c). The title compound, obtained as a white solid (188 mg, 88%). [aD 23 = -0.4* (c = 0.25, chloroform); Rf= 0.45 in (10:90 methanol: chloroform); Mp = 127-130*C; 1 H-NMR (400 MHz, DMSO-d 6 ) 8 6.05 (br s, 2H), 4.17 (dd, J = 11.5, 3.5 Hz, 1H), 4.01 (dd, J = 11.5, 3.5 Hz, 1H), 3.30 (dd, J = 18.5, 13.5 Hz, 8H), 1.68 30 1.59 (m, 1H), 1.53 (m, 1H), 1.42 (m, 2H), 1.31 (d, J = 13.0 Hz, 1H), 1.25-1.12 (m, 1H), 0.95 (m, 2H), 0.87 (d, J = 6.5 Hz, 3H), 0.82 (s, 6H); Mass spectrum (ESI +ve) m/z 312 (MH*).

161 Example 95: (-)-N'-Methyl-N'-(((1R,6S)-2,2,6-trimethylcyclohexyl)methyl) piperazine-1,4-dicarboxamide Sodium triacetoxyborohydride (4.10 g, 19.5 mmol) was added to a stirred solution of (1S,6S)-2,2,6-trimethylcyclohexanecarbaldehyde (1.00 g, 5 6.48 mmol) and methyl amine hydrochloride (1.30 g, 19.5 mmol) in a 10:1 (v/v) mixture of DMF (13 mL) and acetic acid (1.3 mmol). The reaction was stirred at room temperature for 16 hours. The reaction was quenched by adding a saturated solution of sodium carbonate (20 mL) and diluted with distilled water (60 mL). The aqueous solution was extracted with diethyl ether 10 (3 x 75 mL). The combined organic layers were washed with brine (100 mL), dried over magnesium sulfate, filtered and concentrated to give a clear oil (678 mg). The amine (253 mg, 1.50 mmol) was dissolved in anhydrous dichloromethane (20 mL) and cooled to 00C. To this stirred solution was 15 added triphosgene (179 mg, 0.61 mmol) and pyridine (227 mg, 2.88 mmol). The reaction was stirred at 00C until complete consumption of the starting amine by TLC. tert-Butyl piperazine-1-carboxylate (338 mg, 1.81 mmol) was then added to the reaction mixture in one portion. The reaction was stirred at room temperature for 18 hours. The reaction mixture was then diluted with 20 dichloromethane (30 mL) and extracted with 1M HCI (2 x 15 mL) and saturated NaHCO 3 solution (15 mL). The organic layer was washed with brine (20 mL) then dried over sodium sulfate, filtered and concentrated. The intermediate was purified by flash column chromatography (0-50% ethyl acetate in hexanes) and obtained as a clear oil (400 mg, 70%). 25 The Boc-protected intermediate was carried forward following the procedure of Example 71d and was substituted for tert-butyl 4-((E)-3-((1,6 anti)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxylate (71c). The title compound, obtained as a white solid (87.3 mg, 92%). [aD 23 = -7.2* (c = 0.25, chloroform); Rf = 0.35 in (10:90 methanol: chloroform); Mp = 152-157OC; 30 1 H-NMR (400 MHz, DMSO-d 6 ) 8 6.03 (br s, 2H), 3.32-3.19 (m, 5H), 3.09-3.03 (m, 3H), 3.00-2.93 (m, 2H), 2.80 (d, J = 11.5 Hz, 3H), 1.56 (d, J = 12.5 Hz, 162 1H), 1.39 (m, 2H), 1.35-1.26 (m, 2H), 1.17 (m, 1H), 0.99-0.89 (m, 5H), 0.82 (d, J = 6.5 Hz, 3H), 0.77 (s, 3H); Mass spectrum (ESI +ve) m/z 325 (MH*). Example 96: N'-(((1R,6S)-2,2,6-trimethylcyclohexyl)methyl)piperazine 5 1,4-dicarboxamide Hydroxylamine hydrochloride (1.35 g, 19.5 mmol) was dissolved in a 1:1 (v/v) solution of ethanol and water, and treated with sodium bicarbonate (1.64 g, 19.5 mmol) at room temperature for 10 minutes. (1S,6S)-2,2,6 trimethylcyclohexanecarbaldehyde (1.00 g, 6.48 mmol) was added and the 10 reaction mixture was heated to reflux and stirred for 3 days. The reaction was concentrated and the residue dissolved in brine (50 mL) and extracted with chloroform (3 x 25 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated to give the oxime as a clear oil (-1.1 g). 15 The crude oxime (750 mg, 4.43 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL) and cooled to 0*C. To this stirred solution was added lithium aluminum hydride (168 mg, 4.43 mmol), and the reaction was heated to reflux for 18 hours. The reaction slurry was filtered through a pad of Celite and washed with tetrahydrofuran (10 mL). The crude mixture was then 20 concentrated and the residue dissolved in diethyl ether (5 mL). The solution was filtered through a plug of silica gel and eluted with 10% methanol in chloroform (3 x 50 mL). The solution was then concentrated to give the product amine as a colorless oil (200 mg, 29%). The amine (200 mg, 1.29 mmol) was dissolved in anhydrous 25 dichloromethane (20 mL) and cooled to 0*C. To this stirred solution was added triphosgene (573 mg, 1.93 mmol) and triethylamine (522 mg, 5.16 mmol). The reaction was stirred at 0*C until complete consumption of the starting amine by TLC. tert-Butyl piperazine-1-carboxylate (264 mg, 1.42 mmol) was then added to the reaction mixture in one portion. The reaction 30 was stirred at room temperature for 18 hours. The reaction was quenched with a saturated solution of ammonium chloride (30 mL) extracted with 163 dichloromethane (3 x 20 mL). The combined organic layers were washed with brine (20 mL) then dried over magnesium sulfate, filtered and concentrated. The intermediate was purified by flash column chromatography (20-40% ethyl acetate in hexanes) and obtained as a white solid (360 mg, 76%). 5 The Boc-protected intermediate was carried forward following the procedure of Example 71d and was substituted for tert-butyl 4-((E)-3-((1,6 anti)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxylate (71c). The title compound, obtained as a yellow solid (63.0 mg, 74%). Rf = 0.30 in (10:90 methanol: chloroform); Mp = 158*C (decomp); 1 H-NMR (400 MHz, CD 3 0D) 6 10 3.43 (m, 8H), 3.13 (m, 1H), 1.65 (s, 1H), 1.54-1.33 (m, 4H), 1.32-1.23 (m, 1H), 1.05-1.00 (m, 5H), 0.97 (d, J = 6.5 Hz, 3H), 0.87 (s, 3H); Mass spectrum (ESI +ve) m/z 311 (MH*). Example 97: 4-(((1R,6S)-2,2,6-trimethylcyclohexanecarboxamido)methyl) 15 piperidine-1-carboxamide (1S,6S)-2,2,6-trimethylcyclohexanecarbaldehyde (2.75 g, 17.8 mmol) was added dropwise to a solution of 60% nitric acid (1.5 mL) at 55 0 C and stirred for 30 minutes. The reaction was then cooled to room temperature and diluted with water (10 mL) and neutralized with sodium bicarbonate. The 20 aqueous solution was then extracted with dichloromethane (3 x 5 mL). The aqueous layers was then acidified with 1M HCI until pH = 1.0, and extracted with diethyl ether (3 x 10 mL). The combined organic layers were dried over magnesium sulfate, dried and concentrated to obtain the crude acid as a solid (2.00 g, 66%). 25 To a solution of crude acid (170 mg, 1.00 mmol) in acetonitrile (3.0 mL) was added 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU, 418 mg, 1.10 mmol). The solution was stirred at room temperature for 30 minutes then diisopropylethylamine (383 pL, 2.20 mmol) and tert-butyl 4-(aminomethyl)piperidine-1-carboxylate (214 mg, 1.00 30 mmol) was added to the reaction mixture. The reaction was then stirred at room temperature for 16 hours. The reaction was quenched with a 1M 164 solution of hydrochloric acid (2 mL) and the biphasic mixture was separated. The organic layer was concentrated in vacuo (40*C) and the crude material loaded on to silica gel for purification via flash column chromatography running an isocratic eluent of 30% ethyl acetate in hexanes. The title 5 compound was isolated as a white solid (160 mg, 30%). The Boc-protected intermediate was carried forward following the procedure of Example 71d and was substituted for tert-butyl 4-((E)-3-((1,6 anti)-2,2,6-trimethylcyclohexyl)acryloyl) piperazine-1-carboxylate (71c). The title compound, obtained as a white solid (22.0 mg, 24%). Rf = 0.22 in (10:90 10 methanol: chloroform); Mp = 185-187*C; 1 H-NMR (400 MHz, CDCl 3 ) 8 5.61 (br s, 1H), 4.57 (br s, 2H), 3.94 (d, J = 12.5 Hz, 2H), 3.21 (td, J = 12.5, 6.5 Hz, 1H), 3.14-3.03 (td, J = 12.5, 6.5 Hz, 1H), 2.79 (t, J = 12.5 Hz, 2H), 1.86 (m, 1H), 1.71 (m, 4H), 1.55-1.44 (m, 2H), 1.38 (m, 2H), 1.28-1.06 (m, 4H), 1.00 (s, 3H), 0.92 (s, 3H), 0.83 (m, 4H); Mass spectrum (ESI +ve) m/z 310 (MH*). 15 Example 98: (E)-2-(1-(3-(2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl)azetidin-3-yl) acetamide 98a. (E)-Methyl 2-(I-(3-(2,6,6-trimethylcyclohex-1-en-1 20 yl)acryloyl)azetidin-3-yl) acetate The title compound, obtained as a yellow oil (650 mg, 85%), was prepared from the product of Example 71b by following the procedure of Example 3 except methyl 2-(azetidin-3-yl)acetate was substituted for methylamine hydrochloride. Rf = 0.29 in (60:40 ethyl acetate: hexanes); 1

H

25 NMR (400 MHz, DMSO-d 6 ) 8 7.76 (d, J = 15.5 Hz, 1H), 6.42 (d, J = 15.5 Hz, 1H), 4.92 (t, J = 8.5 Hz, 1H), 4.78-4.66 (m, 1H), 4.48 (dd, J = 8.5, 5.5 Hz, 1H), 4.25 (dd, J = 10.5, 5.5 Hz, 1H), 4.17 (s, 3H), 3.52 (td, J = 13.5, 5.5 Hz, 1H), 3.23 (d, J = 7.5 Hz, 2H), 2.57 (t, J = 6.0 Hz, 2H), 2.26 (s, 3H), 2.14 (td, J = 8.5, 6.0 Hz, 2H), 1.99 (dd, J = 7.5, 4.0 Hz, 2H), 1.56 (s, 6H); Mass spectrum (ESI 30 +ve) m/z 306 (MH*).

165 98b. (E)-2-(1-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acryloyl)azetidin-3-yl) acetamide (E)-Methyl 2-(1-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acryloyl)azetidin 3-yl) acetate (100 mg, 0.344 mmol) was dissolved in a solution of 7N 5 ammonia in methanol (2.0 mL) and stirred at room temperature for 2 days. The title compound was purified by preparative plate thin layer chromatography (5:95 methanol: chloroform) to afford a clear oil (45.5 mg, 48%). Rf= 0.60 in (10:90 methanol: chloroform); 'H-NMR (400 MHz, CDCl 3 ) 6 7.28 (d, J = 15.5 Hz, 1H), 5.81 (d, J = 15.5 Hz, 1H), 5.79 (br s, 1H), 5.52 (br s, 10 1H), 4.39 (t, J = 8.5 Hz, 1H), 4.23 (t, J = 9.5 Hz, 1H), 3.90 (dd, J = 8.0, 5.5 Hz, 1H), 3.74 (dd, J = 10.0, 5.5 Hz, 1H), 3.13-2.91 (m, 1H), 2.67-2.47 (m, 2H), 2.03 (t, J = 6.0 Hz, 2H), 1.72 (s, 3H), 1.59 (dd, J = 12.0, 6.90 Hz, 2H), 1.50 1.41 (m, 2H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 291 (MH*). 15 Example 99: (E)-3-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acrylamido) azetidine-1-carboxamide 99a. (E)-tert-Butyl 3-(3-(2,6,6-trimethylcyclohex-1-en-1-yl)acrylamido) azetidine-1 -carboxylate 20 The title compound, obtained as a clear oil (172 mg, 97%), was prepared from the product of Example 1 by following the procedure of Example 3 except tert-butyl 3-aminoazetidine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.40 in (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, DMSO-d 6 ) S 7.10 (s, 1H), 6.49 (d, J = 16.0 Hz, 1H), 5.13 25 (d, J = 16.0 Hz, 1H), 3.80 (m, 1H), 3.42 (m, 2H), 3.01 (m, 2H), 1.28 (t, J = 6.0 Hz, 2H), 0.95 (s, 3H), 0.89-0.80 (m, 2H), 0.70 (m, 2H), 0.63 (s, 9H), 0.26 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 349 (MH*).

166 99b. (E)-3-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acrylamido)azetidine-1 carboxamide The title compound, obtained as a white solid (17 mg, 12%), was prepared from the product of Example 99a by following the procedure of 5 Example 71d. Mp = 185-186*C (decomp); Rf = 0.30 in (10:90 methanol: dichloromethane); 1 H-NMR (400 MHz, MeOD) 8 7.91 (s, 1H), 7.30 (d, J = 16.0 Hz, 1H), 5.96 (d, J = 16.0 Hz, 1H), 4.70-4.59 (m, 1H), 4.25 (t, J = 8.0 Hz, 2H), 3.84 (dd, J = 8.5, 5.50 Hz, 2H), 2.08 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.66 (m, 2H), 1.51 (m, 2H), 1.07 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 292 (MH*). 10 Example 100: (E)-3-(2,6,6-Trimethylcyclohex-1-en-1-yI)-N-(2 ureidoethyl)acrylamide 100a. (E)-tert-Butyl (2-(3-(2,6,6-trimethylcyclohex-1-en-1-yl) acrylamido) ethyl)carbamate 15 The title compound, obtained as a white solid (262 mg, 77%), was prepared from the product of Example 1 by following the procedure of Example 3 except tert-butyl (2-aminoethyl)carbamate was substituted for methylamine hydrochloride. Rf = 0.34 in (50:50 ethyl acetate: hexanes); 1

H

NMR (400 MHz, CDCl 3 ) 8 7.28 (d, J = 15.5 Hz, 1H), 6.29 (br s, 1H), 5.74 (d, J 20 = 15.5 Hz, 1H), 5.01 (br s, 1H), 3.45 (dd, J = 11.0, 5.5 Hz, 2H), 3.32 (d, J = 5.5 Hz, 2H), 2.02 (t, J = 7.0 Hz, 2H), 1.71 (s, 3H), 1.65-1.56 (m, 2H), 1.49 1.38 (m, 11 H), 1.03 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 337 (MH*). 100b. (E)-3-(2,6,6-Trimethylcyclohex-1-en-1-yl)-N-(2 25 ureidoethyl)acrylamide The title compound, obtained as a white solid (40.0 mg, 24%), was prepared from the product of Example 100a by following the procedure of Example 71d. Mp = 165-167*C; Rf = 0.30 in (10:90 methanol: dichloromethane); 1 H-NMR (400 MHz, MeOD) 8 7.29 (d, J = 16.0 Hz, 1H), 30 5.95 (d, J = 16.0 Hz, 1H), 3.44-3.18 (m, 4H), 2.10 (t, J = 6.0 Hz, 2H), 1.78 (s, 167 3H), 1.73-1.61 (m, 2H), 1.53 (m, 2H), 1.09 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 280 (MH*). 100c. (E)-N-(2-(2,2,2-Trifluoroacetamido)ethyl)-3-(2,6,6-trimethylcyclohex 1-en-1-yi) acrylamide 5 The title compound, obtained as a white solid (45.0 mg, 23%), was isolated as a by-product from the above reaction: Example 1 00b. Rf = 0.50 in (10:90 methanol: chloroform); 1 H-NMR (400 MHz, MeOD) 6 7.29 (d, J = 16.0 Hz, 1H), 5.93 (d, J = 16.0 Hz, 1H), 3.46 (s, 4H), 2.10 (t, J = 6.0 Hz, 2H), 1.77 (s, 3H), 1.67 (m, 2H), 1.52 (m, 2H), 1.08 (s, 6H) ppm; 19 F-NMR (376 MHz, 10 CDCl 3 ) 6 -77.4 (s) ppm; Mass spectrum (ESI +ve) m/z 333 (MH*). Example 101: (E)-N-Methyl-N-(2-(1-methylureido)ethyl)-3-(2,6,6 trimethylcyclohex-1-en-1-yl) acrylamide 15 101a. (E)-N-Methyl-N-(2-(methylamino)ethyl)-3-(2,6,6-trimethylcyclohex-l en-1 -yl)acrylamide The title compound, obtained as a white solid (200 mg, 75%), was prepared from the product of Example 1 by following the procedure of Example 3 except N 1

,N

2 -dimethylethane-1,2-diamine (10 equiv) was 20 substituted for methylamine hydrochloride. Rf = 0.20 in (10:90 methanol: dichloromethane); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.31 (d, J = 15.5 Hz, 1H), 6.21 (d, J = 15.5 Hz, 1H), 3.74 (m, 2H), 3.36 (m, 2H), 3.16 (s, 3H), 2.86 (s, 3H), 2.05 (m, 2H), 1.76 (s, 3H), 1.67-1.56 (m, 2H), 1.51-1.42 (m, 2H), 1.05 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 265 (MH*). 25 101b. (E)-N-Methyl-N-(2-(1-methylureido)ethyl)-3-(2,6,6 trimethylcyclohex-1-en-1-yl) acrylamide The title compound, obtained as a white solid (36.0 mg, 26%), was prepared from the product of Example 101a by following the procedure of 30 Example 71d. Mp = 118-120'C; Rf = 0.18 in (5:95 methanol: 168 dichloromethane); 1 H-NMR (400 MHz, CDCl 3 ) 8 7.33 (d, J = 15.5 Hz, 1H), 6.20 (d, J = 15.5 Hz, 1H), 5.01 (br s, 2H), 3.55 (m, 2H), 3.47-3.38 (m, 2H), 3.13 (s, 3H), 2.96 (s, 3H), 2.04 (t, J = 6.0 Hz, 2H), 1.75 (s, 3H), 1.62 (m, 2H), 1.50-1.43 (m, 2H), 1.05 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 308 5 (MH*). Example 102: (E)-4-(3-(2,2,6,6 Tetramethylcyclohexyl)acryloyl)piperazine-1 -carboxamide 10 102a. (E)-Ethyl 3-(2,2,6,6-tetramethylcyclohexyl)acrylate The title compound, obtained as a colorless oil (0.901 g, 80%), was prepared by following the procedure of Example 71a except 2,2,6,6 tetramethylcyclohexanecarbaldehyde was substituted for (1,6-antl)-2,2,6 trimethylcyclohexanecarbaldehyde. Rf= 0.66 (5:95 ethyl acetate: hexanes); H 15 NMR (400 MHz, CDCl 3 ) 8 6.96 (dd, J = 15.5, 11.0 Hz, 1H), 5.77 (d, J = 15.5 Hz, 1H), 4.19 (q, J = 7.0 Hz, 2H), 1.68-1.56 (m, 2H), 1.50-1.43 (m, 3H), 1.30 (t, J = 7.0 Hz, 3H), 1.19-1.10 (m, 2H), 0.97 (s, 6H), 0.79 (m, 6H) ppm; Mass spectrum (ESI +ve) m/z 239 (MH*). 20 102b. (E)-3-(2,2,6,6-Tetramethylcyclohexyl)acrylic acid The title compound, obtained as a white solid (0.720 g, 93%), was prepared from the product of 102a by following the procedure of Example 60c. Mp = 138-140*C; Rf= 0.20 (10:90 ethyl acetate: hexanes); 'H NMR (400 MHz, CDCl 3 ) 8 7.10 (dd, J = 15.5, 11.0 Hz, 1H), 5.80 (d, J = 15.5 Hz, 1H), 1.70 (d, J 25 = 11.0 Hz, 1H), 1.59 (m, 1H), 1.47 (m, 3H), 1.21-1.11 (m, 2H), 0.98 (s, 6H), 0.80 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 211 (MH*). 102c. (E)-tert-Butyl 4-(3-(2,2,6,6 tetramethylcyclohexyl)acryloyl)piperazine-1 -carboxylate The title compound, obtained as a white solid (210 mg, 88%), was 30 prepared from the product of Example 102b by following the procedure of 169 Example 3 except tert-butyl piperazine-1-carboxylate was substituted for methylamine hydrochloride. Rf = 0.40 in (25:75 ethyl acetate: hexanes); 1

H

NMR (400 MHz, -CDCla) 6 6.91 (dd, J = 15.0, 11.0 Hz, 1H), 6.15 (d, J = 15.0 Hz, 1H), 3.73-3.37 (m, 8H), 1.62 (m, 2H), 1.52-1.36 (m, 11H), 1.21-1.06 (m, 5 2H), 0.97 (s, 6H), 0.79 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 379 (MH*). 102d. (E)-4-(3-(2,2,6,6-Tetramethylcyclohexyl)acryloyl)piperazine-l carboxamide The title compound, obtained as a white solid (111 mg, 74%), was prepared from the product of example 102c by following the procedure of 10 Example 71d. Mp = 172-174*C; Rf = 0.34 in (10:90 methanol: chloroform); IH NMR (400 MHz, DMSO-d 6 ) 8 6.65 (dd, J = 15.0, 11.0 Hz, 1H), 6.42 (d, J = 15.0 Hz, 1H), 6.04 (s, 2H), 3.48 (m, 4H), 3.35-3.26 (m, 4H), 1.73 (d, J = 11.0 Hz, 1H), 1.57 (d, J = 13.0 Hz, 1H), 1.42 (m, 3H), 1.14 (m, 2H), 0.91 (s, 6H), 0.76 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 322 (MH*). 15 Example 103: (E)-1 -Morpholino-3-(2,2,6,6-tetramethycyclohexyl)prop- 2 en-1 -one The title compound, obtained as a white solid (42.1 mg, 51%), was prepared from the product of Example 102b by following the procedure of 20 Example 3 except morpholine was substituted for methylamine hydrochloride. Mp = 84-85*C; Rf = 0.35 in (30:70 ethyl acetate: hexanes); 'H-NMR (400 MHz, CDCl 3 ) 6 6.92 (dd, J = 15.0, 11.0 Hz, 1H), 6.13 (d, J = 15.0 Hz, 1H), 3.63 (m, 8H), 1.63 (m, 2H), 1.51-1.40 (m, 3H), 1.15 (m, 2H), 0.97 (s, 6H), 0.79 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 280 (MH*). 25 Example 104: N-((2,6,6-Trimethylcyclohex-1-en-1-yl)methyl)morpholine 4-carboxamide The title compound, obtained as a white solid (337 mg, 77%), was prepared from 2-(2,6,6-trimethylcyclohex-1-en-1-yl)acetic acid by following the 30 procedure of Example 55a except morpholine was substituted for tert-butyl 170 piperazine-1-carboxylate. Mp = 89-91*C; Rf = 0.20 in (25:75 ethyl acetate: hexanes); 1 H-NMR (400 MHz, CDCl 3 ) 5 3.99 (br s, 1H), 3.79 (d, J = 3.5 Hz, 2H), 3.69-3.62 (m, 4H), 3.32-3.25 (m, 4H), 1.93 (t, J = 6.0 Hz, 2H), 1.64 (s, 3H), 1.60-1.53 (m, 3H), 1.45-1.37 (m, 2H), 0.98 (s, 6H) ppm; Mass spectrum 5 (ESI +ve) m/z 267 (MH*). Example 105: (E)-4-(3-(2,6,6-Trimethylcyclohex-1-en-1-yl)acryloyl) piperazine-1 -carbothioamide The product of Example 28b (865 mg, 2.39 mmol) was dissolved in 10 dichloromethane (18 mL) was added dropwise a 4.0 M solution of hydrochloric acid in 1,4-dioxane (6.0 mL, 23.9 mmol). The reaction mixture was stirred at room temperature for 18 hours then concentrated in vacuo. The crude material was dissolved in dichloromethane (100 mL) and extracted with 1M sodium hydroxide (3 x 50 mL) and then washed with brine 15 (50 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give a crude oil. The crude oil was dissolved in tetrahydrofuran (20 mL) and triphenylmethylisothiocyanate (719 mg, 2.39 mmol) was added to the solution. The reaction was heated to reflux for 7 days., and then concentrated to 20 dryness. Purification via preparative plate thin layer chromatography (7:93 methanol: chloroform) afforded the title compound as a white solid (42.0 mg, 5%). Rf= 0.35 (10:90 methanol: dichloromethane); 1 H NMR (400 MHz, CDC 3 ) 8 7.40 (d, J = 15.5 Hz, 1H), 6.17 (d, J = 15.5 Hz, 1H), 5.82 (s, 2H), 4.12 (s, 2H), 3.91-3.67 (m, 6H), 2.04 (d, J = 6.0 Hz, 2H), 1.75 (s, 3H), 1.66-1.59 (m, 25 2H), 1.52-1.42 (m, 2H), 1.05 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 322

(MH*).

171 Example 106: (E)-2-Ethynyl-4-(3-(2,6,6-trimethylcyclohex-1-en-1 yl)acryloyl)piperazine-1 -carboxamide 106a. tert-Butyl 4-tritylpiperazine-1 -ca rboxylate 5 A solution of Boc-piperazine (14.3 g, 77.0 mmol) and triethylamine (11.0 mL, 77.0 mmol) in dichloromethane (300 mL) was stirred at room temperature under argon. To the reaction flask was added trityl chloride (21.5 g, 77.0 mmol) and the reaction mixture was stirred for 1 hour at room temperature. The solution was then washed with saturated ammonium 10 chloride (100 mL), water (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The title compound was isolated as a white solid (11.8 g, 36%). Rf= 0.86 (20:80 ethyl acetate: hexane); 1 H NMR (400 MHz, CDCl 3 ) 6 7.49 (m, 6H), 7.34-7.24 (m, 6H), 7.18 (m, 3H), 3.63-3.49 (m, 4H), 2.57-1.93 (m, 4H), 1.42 (s, 9H) ppm; Mass spectrum (ESI +ve) m/z 329 (MH* - tBu). 15 106b. tert-Butyl 2-formy 1-4-trityl pi perazine-1 -ca rboxylate A solution of the product of Example 106a (7.30 g, 170 mmol) and

N',N',N

2

,N

2 -tetramethylethylene-1,2-diamine (3.90 mL, 26.0 mmol) in anhydrous diethyl ether (250 mL) was stirred at -78 0 C under argon. To this 20 stirred solution was added sec-butyllithium (1.4 M in cyclohexane, 18.5 mL, 26.0 mmol) over 10 minutes. The reaction mixture was stirred for 1 hour at 78 0 C, then DMF (2.00 mL, 26.0 mmol) was added in one portion and the mixture was stirred for 1 hour at -78 0 C. The reaction was quenched by the addition of saturated ammonium 25 chloride (30 mL) at -78 0 C. The solution was vigorously stirred and allowed to warm to room temperature over 40 mintues. The reaction mixture was concentrated in vacuo, and the residue diluted with brine (60 mL) and extracted with chloroform (3 x 100 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated. The title compound was 30 isolated as a white solid (7.42 g, 96%). 1 H NMR displays a mixture of rotamers. Rf= 0.33 (10:90 ethyl acetate: hexane); 1H NMR (400 MHz, CDCla) 172 8 9.85 (m, 2H), 7.45 (m, 12H), 7.36-7.25 (m, 12H), 7.20 (m, 6H), 4.60 (s, 1H), 4.41 (s, 1H), 3.93-3.71 (m, 2H), 3.69-3.32 (m, 4H), 2.98 (m, 2H), 1.98 (s, 2H), 1.56-1.35 (m, 22H) ppm; Mass spectrum (ESI +ve) m/z 401 (MH* - tBu). 5 Example 106c. tert-Butyl 2-ethynyl-4-tritylpiperazine-1-carboxylate The Ohira-Bestmann reagent was prepared in situ by stirring dimethyl (2-oxopropyl) phosphonate (0.720 mL, 5.30 mmol), 4 acetamidobenzenesulfonyl azide (1.30 g, 5.30 mmol) and potassium carbonate (1.82 g, 13.0 mmol) in acetonitrile (60 mL) at room temperature for 10 18 hours. A slurry of the product of Example 106b (2.00 g, 4.40 mmol) in methanol (12 mL) was added and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo, and the residue was taken up in ethyl acetate (150 mL) and washed with brine (100 mL). The 15 aqueous phase was extracted with ethyl acetate (2 x 100 mL). The combined organic phases were washed with saturated sodium bicarbonate (150 mL), brine (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The product was purified by column chromatography (neutral alumina, isocratic elution 40:60 dichloromethane:hexane). The title compound was 20 isolated as a white foam (0.80 g, 40%). Rf = 0.57 (20:80 ethyl acetate: hexane); 1 H NMR (400 MHz, CDCl 3 ) 8 7.90-7.39 (m, 6H), 7.30 (m, 6H), 7.17 (m, 3H), 4.98-4.68 (m, 1H), 3.80-3.73 (m, 1H), 3.63-3.41 (m, 1H), 3.35 (m, 1H), 3.17-2.99 (m, 1H), 2.54 (s, 1H), 1.75-1.64 (m, 1H), 1.42 (s, 9H) ppm; Mass spectrum (ESI +ve) m/z 211 (MH* - trityl). 25 Example 106d. tert-Butyl 2-ethynylpiperazine-1 -carboxylate To a solution of the product of Example 106c (0.350 g, 0.800 mmol) in dichloromethane (5.0 mL) at 00C was added a trichloroacetic acid (2% w/v in dichloromethane, 5.0 mL). The mixture was stirred at 0 0 C for 25 minutes then 30 quenched with aqueous sodium hydroxide (1N, 10 mL). The organic phase was removed and the aqueous phase was extracted with dichloromethane (3 173 x 10 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The product was purified by column chromatography (40 g silica gel, gradient elution 5:95:0.1 methanol: dichloromethane: ammonium hydroxide to 10:90:0.1 methanol: 5 dichloromethane: ammonium hydroxide). This provided the title compound as clear oil (48 mg, 29%). Rf= 0.5 (10:90 ethyl acetate: hexane); Mass spectrum (ESI +ve) m/z 211 (MH*). Example 106e. (E)-tert-Butyl 2-ethynyl-4-(3-(2,6,6-trimethylcyclohex-1-en 10 1-yl)acryloyl) piperazine-1-carboxylate The title compound, obtained as a white solid (60 mg, 68%), was prepared from the product of Example 1 by following the procedure of Example 3 except the product of Example 106d was substituted for methylamine hydrochloride. Rf= 0.32 (30:70 ethyl acetate: hexane); 1 H NMR 15 (400 MHz, CDCl 3 ) 6 7.38 (d, J = 15.5 Hz, 1H), 6.24 (d, J = 15.5 Hz, 1H), 5.13 4.60 (m, 2H), 3.90 (m, 2H), 3.21 (m, 2H), 2.26 (s, 1H), 2.05 (t, J= 6.0 Hz, 2H), 1.75 (s, 3H), 1.62 (m, 2H), 1.35-1.13 (m, 2H), 1.06 (m, 6H) ppm; Mass spectrum (ESI +ve) m/z 387 (MH*). 20 Example 106f. (E)-2-Ethynyl-4-(3-(2,6,6-trimethylcyclohex-1 -en-1 yl)acryloyl) piperazine-1-carboxamide The title compound, obtained as a clear oil (12 mg, 23%), was prepared from the product of Example 106e by following the procedure of Example 71d. The 1 H NMR exhibits a mixture of rotamers. Rf = 0.20 in (10:90 25 methanol: chloroform); 1 H-NMR (400 MHz, CDCl 3 ) 6 7.41-7.31 (m, 1H), 6.29 6.13 (m, 1H), 5.13-4.99 (m, 1H), 4.87 (s, 2H), 4.78-4.64 (m, 1H), 4.16-4.01 (m, 1H), 3.60-3.46 (m, 1H), 3.38 (s, 2H), 2.86-2.73 (m, 1H), 2.32-2.26 (m, 1H), 2.06-2.00 (m, 2H), 1.73 (s, 3H), 1.60 (d, J = 5.54 Hz, 2H), 1.49-1.42 (m, 2H), 1.21 (m, 3H), 1.04 (m, 6H) ppm; Mass spectrum (ESI +ve) m/z 330 (MH*). 30 174 Example 107: (E)-1-morpholino-3-(3,3,6,6-tetramethylcyclohex-1 enyl)prop-2-en-1-one Example 107a. 1,4,4-trimethylcyclohex-2-enol 5 The title compound, obtained as a light yellow oil (41 g, 90%), was prepared from 4,4-dimethylcyclohex-2-enone (40 g, 0.3 mol) according to the procedure of [Dauben, W.; Michno, D.J. Org. Chem. 1977, 42, 682-685]. Rf = 0.5 (5:1 petroleum ether: ethyl acetate) 1 H NMR (400 MHz, CDCl 3 ) 6 5.46 (d, J = 10.0 Hz, 1H), 5.43 (d, J = 10.0 Hz, 1H), 1.73-1.70 (m, 2H), 1.59-1.56 (m, 10 1H), 1.50-1.45 (m, 1H), 1.27 (s, 3H), 1.01 (s, 3H), 0.95 (s, 3H) ppm; Mass spectrum (ESI +ve) m/z 123 (MH-H 2 0)*. Example 107b. 3,6,6-trimethylcyclohex-2-enone The title compound, obtained as a colorless oil (14 g, 35%), was 15 prepared from the product of Example 107a (40 g, 0.3 mol) according to the procedure of [Dauben, W.; Michno, D.J. Org. Chem. 1977, 42, 682-685]. Rf = 0.4 (5:1 petroleum ether: ethyl acetate) 1 H NMR (400 MHz, CDCl 3 ) 6 5.77 (s, 1H), 2.29 (t, J = 6.0 Hz, 2H), 1.93 (s, 3H), 1.80 (d, J = 6.0 Hz, 3H), 1.09 (s, 6H) ppm; Mass spectrum (ESI +ve) m/z 139 (MH)*. 20 Example 107c. 2,2,5,5-Tetramethylcyclohexa none Cul (6.9 g, 36.2 mmol) was added to a dry 250-mL round-bottom flask equipped with a stir bar and sealed under argon with a septum. The flask was evacuated with a vacuum pump and purged with argon. This process was 25 repeated three times. THF (75 mL) was injected and the slurry was cooled to 78 *C, where MeLi (45 mL, 72 mmol) was added dropwise. The mixture was allowed to warm until homogeneous and was recooled to -78 *C, where

BF

3 Et 2 O (8.9 mL, 72 mmol) was added via a syringe. The product of Example 107b (5.0 g, 36.2 mmol) was added neat and the reaction mixture was stirred 30 for 1.5 h. The reaction was quenched with 250 mL of a 10% NH 4 0H/90% saturated NH 4 CI solution and then extracted with ethyl acetate (250 mL), the organic layer was washed with aqueous saturated sodium bicarbonate (50 mL 175 x 2), brine (50 mL), dried over sodium sulfate and concentrated to give a colorless oil (3.5 g) which was a mixture of the product and starting material. The mixture was chromatographed to afford the title compound as a colorless solid (1.5 g 26%). 5 1 H NMR (400 MHz, CDCl 3 ) 6 2.21 (s, 2H), 1.69-1.65 (m, 2H), 1.61-1.57 (m, 2H), 1.09 (s, 6H), 0.94 (s, 6H); 13C NMR (101 MHz, CDCl 3 ) 6 216.36, 51.32, 44.00, 36.89, 36.62, 34.69, 28.5, 25.15; Mass spectrum (ESI +ve) m/z 155 (MH)*. 10 Example 107d. Methyl 3-(1-hydroxy-2,2,5,5 tetramethylcyclohexyl)propiolate To a solution of lithium diisopropyl amide (3.1 mL, 2M in diethyl ether, 6.2 mmol) in THF (5 mL), cooled to -78 OC, was added methyl propiolate (520 mg, 6.2 mmol) in THF (1 mL) dropwise. The mixture was stirred at this 15 temperature for 1 h, and a solution the product of Example 107c (420 mg, 3.0 mmol) in THF (2 mL) was added dropwise. The mixture was stirred at -78 *C for 1h. The mixture was quenched with aqueous ammonium chloride (10 mL), extracted with ethyl acetate (50 mL), washed with sodium bicarbonate (10 mL), brine (10 mL), dried over sodium sulfate, and concentrated. The residue 20 was purified by column chromatography (10:1 petroleum ether: ethyl acetate) to give the title compound as a light yellow oil (600 mg, yield: 40%). 1 H NMR (400 MHz, CDCl 3 ) 6 3.79 (s, 3H), 1.92 (s, 1H), 1.81 (d, J = 14.3 Hz, 1H), 1.70 (d, J = 14.3 Hz, 1H), 1.66 - 1.62 (m, 1H), 1.43 - 1.31 (m, 3H), 1.12 (s, 3H), 1.055 (s, 3H), 1.050 (s, 3H), 1.02 (s, 3H); Mass spectrum (ESI +ve) m/z 221 25 (MH-H 2 0)*. Example 107e. (E)-methyl 3-(1-hydroxy-2,2,5,5 tetramethylcyclohexyl)acrylate The product of Example 107d (589 mg, 2.47 mmol) in THF (10.0 mL) 30 was added to a solution of Red-Al (4.95 mmol, 3.5 M in toluene, 1.4 mL) in THF (8 mL) dropwise at -72 0C (dry ice - ethanol bath) under nitrogen atmosphere. After stirring at the same temperature for 30 min, the mixture was quenched with 0.1 M HCI (5 mL), extracted with ethyl acetate (50 mL), 176 washed with 0.1 M HCI (20 mL x 3), aqueous sodium bicarbonate (20 mL) and brine (20 mL), dried over sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by column chromatography (10:1 petroleum ether:ethyl acetate) to give the desired product as white solid (380 5 mg, yield: 64%). 'H NMR (400 MHz, DMSO-d 6 ) 6 7.10 (d, J = 15.6 Hz, 1H), 6.07 (d, J = 15.6 Hz, 1H), 3.76 (s, 3H), 1.88 (td, J = 13.6, 4.0 Hz, 1H), 1.73 (d, J = 14.6 Hz, 1 H), 1.48 (td, J = 13.6, 4.0 Hz, 1 H), 1.34 (td, J = 4.0, 2.0 Hz, 1 H), 1.31 (s, 1H), 1.25 (dd, J = 14.6, 1.9 Hz, 1H), 1.17 (dt, J = 13.6, 4.0 Hz, 1H), 1.11 (s, 3H), 0.98 (s, 3H), 0.94 (s, 3H), 0.89 (s, 3H); Mass spectrum (ESI +ve) 10 m/z 241 (MH)*. Example 107f. (E)-methyl 3-(3,3,6,6-tetramethylcyclohex-1-enyl)acrylate To a solution of the compound of Example 107e (380 mg, 1.58 mmol) in acetic acid (1.8 mL) was added acetic anhydride (0.6 mL) followed by 15 acetyl chloride (0.6 mL). The mixture was refluxed for 2h. The reaction solution was concentrated. The residue was taken up in ethyl acetate (50 mL), washed with aqueous sodium bicarbonate (30 mL x 3) and brine (30 mL), dried over sodium sulfate and concentrated. The crude product was purified by column chromatography (10:1 petroleum ether:ethyl acetate) to give the 20 title compound as light yellow oil (230 mg, yield: 65%). 'H NMR (400 MHz, MeOD) 6 7.33 (d, J = 15.8 Hz, 1H), 6.03 (d, J = 15.8 Hz, 1H), 5.78 (s, 1H), 3.76 (s, 3H), 1.57 - 1.51 (m, 2H), 1.51 - 1.45 (m, 2H), 1.11 (s, 6H), 1.02 (s, 6H); Mass spectrum (ESI +ve) m/z 223 (MH)*. 25 Example 107g. (E)-3-(3,3,6,6-tetramethylcyclohex-1-enyl)acrylic acid To a solution of the compound of Example 107f (150 mg, 0.67 mmol) in methanol (5 mL) and water (1 mL) was added sodium hydroxide (80 mg, 2.0 mmol). The mixture was refluxed for 2h. The reaction solution was concentrated, acidified to ph - 2-3, extracted with ethyl acetate (50 mL), dried 30 over sodium sulfate, and concentrated. The residue was dried under vacuum to give the title compound as colorless oil (130 mg, yield: 93%). The crude compound was used in the next step without further purification. 1H NMR 177 (400 MHz, CDC 3 ) 6 7.42 (d, J = 15.8 Hz, 1H), 6.04 (d, J = 15.8 Hz, 1H), 5.86 (s, 1H), 1.58 - 1.52 (m, 2H), 1.52 - 1.45 (m, 2H), 1.12 (s, 6H), 1.04 (s, 6H). Example 107h. (E)-1-morpholino-3-(3,3,6,6-tetramethylcyclohex-1 5 enyl)prop-2-en-1-one A solution of the compound of Example 107g (60 mg, 0.29 mmol), HATU (165 mg, 0.435 mmol), diisopropylethyl amine (112 mg, 0.87 mmol) in DMF (2 mL) was stirred at rt for 0.5 h. Morpholine (25 mg, 0.29 mmol) was added. The mixture was stirred at rt for 3h. After dilution with water (10 mL), 10 the mixture was extracted with ethyl acetate (50 mL), washed with water (20 mL x 2), brine (20 mL x 2), dried sodium sulfate, and concentrated. The residue was purified by column chromatography (2:1 petroleum ether; ethyl acetate) to give the title compound as a colorless syrup (65 mg, yield: 81%). Preparative HPLC gave 30 mg of the title product as a white solid (30 mg). 15 Further purification by prep-HPLC gave 8 mg of the pure title compound as light yellow solid, 1 H NMR (400 MHz, CDCI 3 ) 6 7.36 (d, J = 15.1 Hz, 1H), 6.41 (d, J = 15.1 Hz, 1H), 5.71 (s, 1H), 3.79 - 3.54 (m, 8H), 1.57 - 1.51 (m, 2H), 1.51 - 1.45 (m, 2H), 1.10 (s, 6H), 1.03 (s, 6H); Mass spectrum (ESI +ve) m/z 278 (MH)*. 20 Example 108 (E)-4-(3-(3,3,6,6-tetramethylcyclohex-1-enyl)acryloyl) piperazine-1 -carboxamide Example 108a. Tert-Butyl 4-carbamoylpiperazine-1-carboxylate 25 To a solution of tert-butyl piperazine-1-carboxylate (5.0 g, 26.8 mmol) in acetic acid (15 mL) and water (25 mL) was added a solution of potassium cyanate (11.25 g, 138.9 mmol) in water (25 mL) dropwise. After addition the mixture was stirred at rt for 4h, during which time a solid precipitated. The solid was collected by filtration, re-dissolved in dichloromethane (20 mL), 30 dried oversodium sulfate, and filtered. The filtrate was concentrated to give the title compound as a white solid (3.3 g, yield: 53%), which was used in the next step without further purification. 1 H NMR (400 MHz, DMSO-d 6 ) 6 6.04 (s, 2H), 3.26 (s, 8H), 1.41 (s, 9H).

178 Example 108b. Piperazine-1-carboxamide trifluoroacetate salt A solution of the product of Example 108a (1.5 g, 6.5 mmol) in trifluoroacetic acid (5 mL) and dichloromethane (15 mL) was stirred at rt for 3h. The mixture was concentrated. The residue was triturated with ethyl 5 acetate (5 mL x 2) and diethyl ether (5 mL x 2), dried under vaccum to give the title compound as colorless syrup (1.5 g, yield: 95%). 1 H NMR (400 MHz, DMSO-d 6 ) 6 9.11 (s, 2H), 7.04 - 5.66 (br, s, 2H), 3.57 - 3.45 (m, 4H), 3.06 (s, 4H). 10 Example 108c. (E)-4-(3-(3,3,6,6-tetramethylcyclohex-1-enyl)acryloyl) piperazine-1 -carboxamide A solution of the product of Example 107g (65 mg, 0.31 mmol), HATU (178 mg, 0.47 mmol), diisopropylethyl amine (120 mg, 0.93 mmol) in DMF (2 mL) was stirred at rt for 0.5 h. The product of Example 108b (75 mg, 0.31 15 mmol) was added. The mixture was stirred at rt for 3h. After dilution with water (10 mL), the mixture was extracted with ethyl acetate (50 mL), washed with water (20 mL x 2), brine (20 mL x 2), driedover sodium sulfate, and concentrated. The residue was purified by column chromatography (10:1 dichloromethane:methanol) to afford the title compound as a white solid (70 20 mg, yield:70%). Further purification by prep-HPLC gave a white solid (23 mg, 23%). 1 H NMR (400 MHz, DMSO-d 6 ) 6 7.37 (d, J = 15.0 Hz, 1H), 6.42 (d, J = 14.8 Hz, 1H), 5.74 (s, 1H), 4.59 (s, 2H), 3.90 - 3.32 (m, 8H), 1.58 - 1.52 (m, 2H), 1.52 - 1.46 (m, 2H), 1.10 (s, 6H), 1.04 (s, 6H); Mass spectrum (ESI +ve) m/z 320 (MH)*. 25 Example 109 (E)-4-(3-(3,6,6-trimethylcyclohex-1-enyl)acryloyl)piperazine 1-carboxamide Example 109a. 2,5,5-Trimethylcyclohexanone 30 To a solution of the product of Example 107b (1.0 g, 7.24 mmol) in methanol (20 mL) was added Pd/C (0.2 g). The mixture was stirred at 25 0 C under a hydrogen atmosphere overnight. The reaction mixture was filtered through Celite and the filtrate was concentrated under reduced pressure to 179 give a colorless oil which was purified by chromatography to give the title compound as colorless oil (300 mg, 30%). Rf = 0.6 (5:1 petroleum ether:ethyl acetate); 1 H NMR (400 MHz, CDCl 3 ) 6 5.77 (s, 1H), 2.29 (t, J = 6.0 Hz, 2H), 1.93 (s, 3H), 1.80 (d, J = 6.0 Hz, 3H), 1.09 (s, 6H); 13 CNMR (100 MHz, CDCl 3 ) 5 6 215.49, 46.21, 44.09, 39.53, 34.72 , 29.70, 25.02, 24.95, 21.89; Mass spectrum (ESI +ve) m/z 141 (MH)*. Example 109b. Methyl 3-(1 -hydroxy-2,2,5-trimethylcyclohexyl)propiolate To a solution of methyl propiolate (0.6 g, 7.13 mmol) in THF (12 mL), 10 cooled to -78 0 C, was added dropwise lithium diisopropyl amide solution (3.6 mL, 2M in ether, 7.13 mmol). The mixture was stirred at this temperature for 1h, and a solution of the product of Example 109a (1.0 g, 7.13 mmol) in THF (12 mL) was added dropwise. The mixture was stirred at -78 *C for 1h. The mixture was quenched with ammonium chloride (aq. 5 mL), extracted with 15 ethyl acetate (30 mL), washed with sodium bicarbonate (aq. 10 mL), brine (10 mL), dried over sodium sulfate and concentrated. The residue was purified by column chromatography (10:1 petroleum ether:ethyl acetate) to give a light yellow oil (0.76 g, 48%).'H NMR (400 MHz, CDCl 3 ) 6 3.80 (s, 3H), 2.04 (s, 1H), 1.86 - 1.77 (m, 2H), 1.66 - 1.57 (m, 2H), 1.55 - 1.48 (m, 1H), 1.44 20 1.37 (m, 1H), 1.28 (t, J = 7.1 Hz, 1H), 1.16 - 1.12 (s, 3H), 0.99 (s, 3H), 0.97 (d, J = 6.3 Hz, 3H). Example 109c. (E)-methyl 3-(1-hydroxy-2,2,5 tri methy Icyc lohexy I)a cry late 25 To a solution of Red-Al (1.9 mL, 6.7 mmol) in THF (11 mL) under argon, cooled to -72 0 C, was added dropwise the product of Example 109b (0.75 g, 3.35 mmol) in THF (14 mL). The mixture was stirred at this temperature for 1h. The mixture was quenched with 0.1 M HCI (150 mL). The solution was concentrated under reduced pressure and was then diluted with 30 ethyl acetate (60 mL). The mixture was separated and the aqueous layer was extracted with ethyl acetate (60 mL). The combined organic layers were then washed with sat. sodium bicarbonate (15 mL), dried over sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by 180 column chromatography (10:1 petroleum ether;ethyl acetate) to give the title compound as light yellow oil (0.55 g, 73%). 1 H NMR (400 MHz, CDCl 3 ) 6 7.45 (d, J= 15.5 Hz, 1H), 6.17 (d, J= 15.5 Hz, 1H), 3.77 (s, 3H), 1.68 (m, 1H),1.65 (s, 2H), 1.60 (d, J = 1.9 Hz, 1H), 1.57 (t, J = 3.4 Hz, 1H), 1.51 (d, J = 3.0 Hz, 5 1H), 1.46 (dd, J = 4.2, 2.1 Hz, 1H), 1.43 (d, J = 3.9 Hz, 1H), 1.28 - 1.19 (m, 3H), 1.03 (s, 3H), 0.94 (d, J = 7.9 Hz, 3H), 0.85 (s, 3H); Mass spectrum (ESI +ve) m/z 209 (MH-H 2 0)*. Example 109d. (E)-methyl 3-(3,6,6-trimethylcyclohex-1-enyl)acrylate 10 To a solution of the product of Example 109c (120 mg, 0.53 mmol) in carbon tetrachloride (5 mL) at 0 0C was added a solution of Martin's sulfurane (0.9 g, 1.33 mmol) in carbon tetrachloride (7.5 mL) under argon. After addition, the cooling bath was removed and the mixture was stirred at rt for 1.5 h. Crushed ice and water (15 mL) were added and after being stirred for 15 20 min, the mixture was extracted with dichloromethane (50 mL). The organic phases were washed with water (5 mL) and brine (5 mL), dried over sodium sulfate and concentrated to dryness. The residue was purified by preparative thin layer chromatography to afford the title compound as colorless oil (75 mg, 68%). 1 H NMR (400 MHz, CDCl 3 ) 6 7.22 (d, J = 15.8 Hz, 1H), 6.00 (d, J = 20 15.8 Hz, 1H), 5.89 (d, J = 2.9 Hz, 1H), 3.70 (s, 3H), 2.22 (m, 1H), 1.76 - 1.65 (m, 1H), 1.61 (s, 1H), 1.58 - 1.46 (m, 1H), 1.49 - 1.37 (m, 1H), 1.30 - 1.17 (m, 1H), 1.07 (d, J = 2.7 Hz, 6H), 0.98 (d, J = 6.7 Hz, 3H). Example 109e. (E)-3-(3,6,6-trimethylcyclohex-1-enyl)acrylic acid 25 To a solution of the product of Example 109d (200 mg, 0.96 mmol) in methanol (7 mL) and water (1 mL) was added sodium hydroxide (115 mg, 2.88 mmol). The mixture was refluxed for 2 h. The reaction mixture was concentrated under reduced pressure and then it was diluted with water (5 mL). 3N HCI was added to adjust the pH to 2. The aqueous layer was 30 extracted with ethyl acetate (3 x 10 mL), washed with brine (5 mL), dried over sodium sulfate and concentrated under reduced pressure to give the crude title product as a brown oil. (170 mg) which was carried on without further purification. 1 H NMR (400 MHz, DMSO) 6 12.25 (s, 1H), 7.16 (d, J = 15.9 Hz, 181 1 H), 6.05 - 5.93 (m, 2H), 2.25 (d, J = 7.3 Hz, 1 H), 1.75 - 1.65 (m, 1 H), 1.57 1.51 (m, 1H), 1.47 - 1.38 (m, 1H), 1.26 - 1.18 (m, 1H), 1.06 (d, J = 6.4 Hz, 6H), 0.98 (d, J= 7.2 Hz, 3H); Mass spectrum (ESI +ve) m/z 195 (MH)*. 5 Example 109f. (E)-4-(3-(3,6,6-trimethylcyclohex-1 enyl)acryloyl)piperazine-1 -carboxamide To a solution of the product of Example 109e (85 mg, 0.44 mmol) and HATU (250 mg, 0.66 mmol) in DMF (3 mL) was added the product of Example 108b (107 mg, 0.44 mmol) followed by diisopropylethyl amine (171 mg, 1.32 10 mmol). The mixture was stirred at rt for 2 h. Water (20 mL) was added to the reaction mixture and it was extracted with ethyl acetate (3 x 10 mL). The organic layer was washed with brine (5 mL), dried over sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by preperative thin layer chromatography to give a colorless syrup. Further 15 purification by column chromatography (20:1 dichloromethane:methanol) gave the title compound as a white solid. (36 mg, 27%). 'H NMR (400 MHz, CDC 3 ) 6 7.38 (d, J = 15.2 Hz, 1H), 6.44 (d, J = 15.1 Hz, 1H), 5.88 (d, J = 2.8 Hz, 1H), 4.60 (s, 2H), 3.71 (d, J = 26.5 Hz, 4H), 3.52 (s, 4H), 2.26 (d, J = 7.5 Hz, 1H), 1.82 - 1.68 (m, 1H), 1.58 (ddd, J = 13.1, 6.0, 3.0 Hz, 1H), 1.54 - 1.44 (m, 1H), 20 1.34 - 1.20 (m, 1H), 1.11 (d, J = 5.1 Hz, 6H), 1.05 (d, J = 7.1 Hz, 3H); Mass spectrum (ESI +ve) m/z 306 (MH)*. Example 110. (E)-1-morpholino-3-(3,6,6-trimethylcyclohex-l-enyl)prop-2 en-I -one 25 To a solution of the product of Example 109e (90 mg, 0.46 mmol) and HATU (266 mg, 0.7 mmol) in DMF (2 mL) was added morpholine (40 mg, 0.46 mmol) followed by diisopropylethyl amine (178 mg, 1.38 mmol). The mixture was stirred at rt for 2 h. Water (20 mL) was added to the reaction mixture and it was extracted with ethyl acetate (3 x 10 mL). The organic layer was washed 30 with brine (5 mL), dried over sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by preparative thin layer chromatography and then column chromatography (2:1 petroleum ether;ethyl 182 acetate) to give the semi pure title compound as a light yellow oil (85 mg). Preparative HPLC gave 36 mg of the desired product as colorless syrup (36 mg). Further purification by column chromatography (2:1 petroleum ether:ethyl acetate) gave the title compound (10 mg, 8%) 1 H NMR (400 MHz, 5 CDCl 3 ) 6 7.37 (d, J = 15.2 Hz, 1H), 6.43 (d, J = 15.1 Hz, 1H), 5.86 (d, J = 2.8 Hz, 1H), 3.68 (d, J = 34.3 Hz, 8H), 2.25 (d, J = 7.1 Hz, 1H), 1.74 (ddd, J = 13.2, 6.3, 3.0 Hz, 1H), 1.62 - 1.40 (m, 1H), 1.33 - 1.16 (m, 2H), 1.10 (t, J = 6.7 Hz, 6H), 1.03 (d, J = 7.1 Hz, 3H); Mass spectrum (ESI +ve) m/z 264 (MH)*. 10 Biology Examples In carrying out the procedures of the present invention it is of course to 15 be understood that reference to particular buffers, media, reagents, cells, culture conditions and the like are not intended to be limiting, but are to be read so as to include all related materials that one of ordinary skill in the art would recognize as being of interest or value in the particular context in which that discussion is presented. For example, it is often possible to substitute one 20 buffer system or culture medium for another and still achieve similar, if not identical, results. Those of skill in the art will have sufficient knowledge of such systems and methodologies so as to be able, without undue experimentation, to make such substitutions as will optimally serve their purposes in using the methods and procedures disclosed herein. 25 The invention is described in more detail in the following non-limiting examples. It is to be understood that these particular methods and examples in no way limit the invention to the embodiments described herein and that other embodiments and uses will no doubt suggest themselves to those skilled in the 30 art.

183 Reagents Monoclonal anti-rhodopsin 1D4 antibody can be purchased from University of British Columbia. 5 Cell lines and culture conditions Stable cell lines expressing opsin protein were generated using the Flp-In T-Rex system. The stable cells were grown in DMEM high glucose 10 media supplemented with 10% (vlv) fetal bovine serum, antibiotic/antimycotic solution, 5 /ml blasticidin and hygromycin at 37 0 C in presence of 5% CO 2 . For all the experiments the cells were allowed to reach confluence and were induced to produce opsin with 1 ig/ml tetracycline after change of media and then compounds were added. The plates were incubated for 48 hours after 15 which the cells were harvested. SDS-PAGE and western blotting 20 Proteins were separated on SDS-PAGE gels and western blotted as described in (Noorwez et al., J. Biol. Chem. 279,16278-16284 (2004)). The in vivo efficacy of the compounds of the invention in treating macular degeneration can be demonstrated by various tests well known in the 25 art. For example, human patients are selected based on a diagnosis of macular degeneration (such as where there is a gross diagnosis of this condition or where they have been shown to exhibit build-up of toxic visual cycle products, such as A2E, lipofuscin, or drusen in their eyes. A compound of the invention, such as that of Formula I and/or Formula 11, is administered 30 to a test group while a placebo, such as PBS or DMSO, is administered to a control group that may be as large or may be somewhat smaller than the test group. The test compound is administered either on a one time basis or on a 184 sequential basis (for example, weekly or daily) or according to some other predeteremined schedule Administration of the test compound is normally by oral or parenteral 5 means and in an amount effective to retard the development and/or reoccurrence of macular degeneration. An effective dose amount is generally in the range of about 1 to 5,000 mg or in the range of 10 to 2,000 mg/kg. Administration may include multiple doses per day. 10 Efficacy of the test compound in retarding progression of macular degeneration is generally by measuring increase in visual acuity (for example, using Early Treatment Diabetic RP Study (ETDRS) charts (Lighthouse, Long Island, N.Y.). Other means of following and evaluating efficacy is by measuring/monitoring the autofluorescence or absorption spectra of such 15 indicators as N-retinylidene-phosphatidylethanolamine, dihydro-N retinylidene-N-retinyl-phosphatidylethanolamine, N-retinylidene-N-retinyl phosphatidylethanolamine, dihydro-N-retinylidene-N-retinyl-ethanolamine, and/or N-retinylidene-phosphatidylethanolamine in the eye of the patient. Autofluorescence is monitored using different types of instrument, for 20 example, a confocal scanning laser ophthalmoscope. Accumulation of lipofuscin in the retinal pigment epithelium (RPE) is a common pathological feature observed in various degenerative diseases of the retina. A toxic vitamin A-based fluorophore (A2E) present within lipofuscin 25 granules has been implicated in death of RPE and photoreceptor cells. Such experiments can employ an animal model which manifests accelerated lipofuscin accumulation to evaluate the efficacy of a therapeutic approach based upon reduction of serum vitamin A (retinol). Administration of test compound to mice harboring a null mutation in the Stargardt's disease gene 30 (ABCA4) produces reductions in serum retinol/retinol binding protein and arrested accumulation of A2E and lipofuscin autofluorescence in the RPE. Test animals are available for use in testing efficacy of a test 185 compound in reducing build-up of toxic pigments, such as lipofuscin. For example, mice have been produced that exhibit increased production of sich toxic product. Such mice have been described in the literature (see, for example, Widder et al., U.S. Pub. 2006/0167088) and their value and utility 5 are well known to those in the art. Showing the efficacy of compounds of the invention in protecting against light toxicity is conveniently performed by methods well known in the art (see, for example, Sieving et al, PNAS, Vol. 98, pp 1835-40 (2001)). 10 Biology Example 1 SDS-PAGE and western blotting 15 Proteins were separated on SDS-PAGE gels and western blotted as described in (Noorwez et al., J. Biol. Chem. 279,16278-16284 (2004)). HEK P23H cell opsin expression was induced as described above for 16 to 24 hrs in the presence of DMSO (blank) or various concentrations of test compound (generally 1 to 40 pM). After incubation cells were lysed in cold Phosphate 20 Buffered Saline with 1% docecyl maltoside (PBD-D) for 1 hr, and the lysate cleared by centrifugation. Total protein (~10 ptg) was loaded on 4-20% SDS polyacrylamide gels (BioRad) and total opsin quantified by western blotting using the anti-rhodopsin monoclonal antibody 1 D4 (2.5 ptg /mL) as the primary antibody and IRDye-labeled goat anti-mouse (Licor) as the secondary 25 antibody for detection. The blots were scanned and opsin levels quantified using the Odyssey infrared scanner and software (Licor Biosystems). Here, a test compound is added to a selected final concentration (20 pM results are reported in Table 1) . The results were calculated as the % of 30 mature P23H opsin (- 52kDA) produced in the HEK 293 cells relative to the control 9-cis retinal at 20 uM defined as producing a 100% response 186 Table 1. Activity in the western blot assay: Compound No. Increase in Mature P23H Opsin 6 113% @ 20pM 14 144% @ 20pjM 15 113% @ 20pjM 17 132% @ 20ptM 21 89% @ 20pjM 29 97% @ 20pjM 34 155% @ 20pjM 37 136% @ 20ptM 44 136% @ 20pjM 45 122% @ 20pjM The western blot results show the total amount of opsin protein 5 produced (as quantified on the gel). A 52kDA band is the fully maturated protein. Rhodopsin generation data then allows determination as to whether it is suitably folded to form rhodopsin when exposed to retinal. Data has shown that not all mature protein is necessarily folded to accept retinal and form pigment but the mutant protein in the presence of chaperone does appear to 10 traffic normally out of the edoplasmic reticulum. Biology Example 2 Rhodopsin Purification and Regeneration 15 P23H cell opsin expression is induced 48 hrs in the presence of DMSO (blank) or various concentrations of test compound (generally 1 to 40 piM). P23H opsin producing cells are washed with PBS and lysed in cold PBS-D for 1 hour. The lysate is cleared by centrifugation and added to 1D4-coupled 20 sepharose beads and incubated for 1 hour at 4 0 C. Opsin was eluted from the antibody beads with a competing peptide corresponding to the last 18 amino 187 acids of rhodopsin in the same buffer. The purified opsin is immediately used for rhodopsin regeneration studies using 9-cis retinal as chromophore. Opsin (=25 pM) is mixed with 10 pM 9-cis retinal and the absorbance is determined over the range of 250-650 nm every two minutes in a Cary 50 5 spectrophotometer (Varian) until no more rhodopsin is regenerated as measured by the increase in 480-500 nm absorbance. Figures 2-17 are the spectral results using selected compounds according to Biology Example 2. Other Embodiments 10 From the foregoing description, it will be apparent that variations and modifications may be made to the invention described herein to adopt it to various usages and conditions. Such embodiments are also within the scope of the following claims. 15 The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in 20 combination with any other embodiments or portions thereof. All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication was specifically and individually indicated to be incorporated 25 by reference.

Claims (76)

1. A compound having the structure of Formula I 5 A-B-Q-V Formula I wherein A is: 10 R 1 R 2 R 1 R 2 R4 Rd Rd R5 R24 Z Rc R3 RC* R3 R6 Ra Rb Ra Rb R7 R 12 .N R R 10 NH 15 R9 0 R R14 R12 Rz T 5R17 R Ri6 R15 T 189 R19 E R20RDNs2 ZT R 2 0 R R 2 R9 R 61, R o T R R 1 6 or R 1 R 2 R9 5 B is -(CH 2 )n-, -CH=CH-, -CH 2 -N(R 22 )-, -CH 2 -0-, -C(O)-CH 2 -C(O) N-E ,or -C(O)NR 22 - , wherein n = 0, 1 or 2, 10 E is -N(R 22 )- or oxygen; Q is -C(O)- , -(CH 2 )a-, -S(0 2 )- or -CH 2 -C(O)- , wherein a is 1 or 2; 2R25 -N Y YY 15 V is NR 21 R 22 , R 23 X, 190 NN R2Y 0 , or R22 N Q'U R 2 3 ,wherein b is 1 or 2 and a is 1 or 2; 5 Y is NR 2 2 , N-Q-U , CR 22 R 23 , oxygen, S(O)n, N-C(S)-NR 2 2 R 2 3 ; N-(C=N-CN)-NR 22 R 23 , N-(C=N-SO 2 CH 3 )-NR 22 R 23 C=NOR 2 2 , C=N-NR 22 R 2 3 or C-Q-U, n is 0, 1 or 2; U is NR 22 R 23 , lower alkyl, haloalkyl, alkoxy, OR 22 or hydrogen; 10 X is hydrogen, alkyl or -C=CR 9 ; R 1 and R 2 are independently -CH 3 or -CH 2 CH 3 ; 15 R 3 is hydrogen, -CH 3 or -CH 2 CH 3 ; Ra, and Rb, are each independently hydrogen, deutero or -CH 3 Re, and Rd, are each independently hydrogen, alkoxy, lower alkyl or 20 alkenyl; R 4 is -CH 3 , -CF 3 , -C 2 H 5 or -C 3 H 5 ; R 5 , R 6 and R 7 are each independently hydrogen, lower alkyl, halogen, dialkylamine, nitro or dialkylamine; 25 Z is CR 3 , CH or nitrogen; R 8 is -CH 2 - or -C(O)-; 191 R 9 , R 14 and R 16 are each independently hydrogen, or -CH 3 ; R 10 is N-R 13 , sulfur or oxygen; 5 R" is =N- , or =C(CH 3 )-; R 12 is lower alkyl, alkoxy or haloalkyl; R 13 is phenyl, lower alkyl or haloalkyl; 10 R 15 is hydrogen or -C(O)CH 3 ; R 17 and R 18 together are -(CH 2 ) 4 - or -CH=CH-CH=CH 15 R 19 and R 20 together are -CH 2 -C(CH 3 ) 2 -CH 2 -C(O)- or -CH=CH-CH=CH-; R 2 1 is hydrogen, -C(O)CH 3 , -CH 3 or -CH 2 CH 3 ; 20 R 2 2 and R 2 3 are each independently hydrogen; or lower alkyl; R 24 and R 25 are each independently hydrogen or -CH 3 ; R 26 is NR 22 R 23 or alkoxy; 25 And wherein R 1 and R 2 taken together or Ra and Rb taken together along with the carbon to which they are attached are cyclopropyl; 30 R 24 and R 25 taken together along with the two carbons to which they are attached are cyclopropyl: R 24 and R 25 taken together is oxo; 192 And wherein T is oxygen, -N(R 1)- or sulfur; E is oxygen, -N(R' 6 )- , sulfur or -C(O)-; 5 including pharmaceutically acceptable salts, solvates and hydrates thereof.

2. The compound of claim 1, wherein A is R 1 R 2 R 1 R 2 Rd Rd I R24 Re R3 Re R3 10 Ra Rb or Ra Rb

3. The compound of claim 2, wherein each of R' ,R 2 and R 3 is methyl.

4. The compound of claim 2, wherein each of R 24 is hydrogen. 15

5. The compound of claim 2, wherein Ra and Rb are each independently hydrogen or methyl.

6. The compound of claim 2, wherein Ra and Rb are each methyl. 20

7. The compound of claim 2, wherein Re and Rd are each independently hydrogen or lower alkyl.

8. The compound of claim 2, wherein Re and Rd are each lower alkyl. 25

9. The compound of claim 2, wherein Re and Rd are each hydrogen.

10. The compound of claim 1, wherein V is: 193 R 25 -N Y R 23 X ;

11. The compound of claim 10, wherein a and b are each 1, X is hydrogen, Y is C-C(O)NR 22 R" or N-C(O)NR 22 R 2 1 wherein R 2 1, R 24 and R 25 5 are all hydrogen.

12. The compound of claim 1, wherein B is CH=CH.

13. The compound of claim 1, wherein Q is -C(O)-. 10

14. The compound of claim 1, wherein X is H, lower alkyl or -CECR 9 .

15. The compound of claim 1, wherein X is H or methyl. 15

16. The compound of claim 14, wherein X is -C=CR 9 .

17. The compound of claim 16, wherein R 9 is hydrogen or methyl.

18. The compound of claim 1, wherein Y is 0 or N-C(O)-NR 22 R. 20

19. The compound of claim 18, wherein Y is N-C(O)-NR 22 R.

20. The compound of claim 19, wherein R 22 and R 23 are each hydrogen. 25

21. A compound having the structure of Formula 11: 194 A-B-Q-N2 P W Formula II 5 W is -OR 2 2 , -NR 22 R 2 3 , -N(R 2 2 )-C(O)-NR 2 2 R 2 3 , -O-C(O)-NR 2 2 R 2 3 , -N(R 22 )-C(S)-NR 22 R 23 , -O-C(S)-NR 2 2 R 2 3 , -S-C(O)-NR 2 2 R 2 3 , -N(R 22 )-(C=N-CN)-NR 2 2 R 2 3 , -N(R 22 )-(C=N-SO 2 Me)-NR 22 R23, or -C(O)N(R9)N(R144)(R 16); 10 wherein p is 0 or 1; and wherein R 9 , R 14 , R 16 , R 22 , R 2 3 , A, B and Q are as defined in claim 1, including pharmaceutically acceptable salts, solvates and hydrates thereof. 15

22. The compound of claim 21, wherein A is: RI R 2 RI R 2 Rd Rd R 24 Rc R 3 Rc R 3 Ra Rb or Ra Rb 20

23. The compound of claim 22, wherein at least one of R 1 , R 2 and R 3 is methyl or ethyl.

24. The compound of claim 23, wherein at least one of R 1 , R 2 and R 3 is 25 methyl.

25. The compound of claim 23, wherein each of R 1 , R 2 and R 3 is 195 methyl.

26. The compound of claim 22, wherein Ra and Rb are each independently hydrogen, deutero or methyl. 5

27. The compound of claim 22, wherein Ra or Rb are each hydrogen.

28. The compound of claim 22, wherein Re and Rd are each independently hydrogen, alkoxy or lower alkyl. 10

29. The compound of claim 28, wherein Re and Rd are each independently hydrogen or methyl.

30. The compound of claim 28, wherein Re and Rd are each hydrogen. 15

31. The compound of claim 21, wherein B is -CH=CH or -CH 2 -CH 2 -.

32. The compound of claim 31, wherein B is -CH=CH-. 20

33. The compound of claim 21, wherein Q is -C(0)-.

34. The compound of claim 21, wherein p is 1.

35. The compound of claim 34, wherein W is -N(R 22 -C(O)-NR 2 2 R 2 3 . 25

36. The compound of claim 34, wherein R 9 , R 22 and R 2 3 are each H.

37. A compound having the structure of compound 33, 52, 63, 71, 72, 73 or 80 including pharmaceutically acceptable salts, solvates and hydrates 30 thereof.

38. A compound having the structure of compound 6, 48, 50, 51, 57, 58, 59, 62 or 106 including pharmaceutically acceptable salts, solvates and 196 hydrates thereof.

39. A compound having the structure of compound 13, 37, 53 or 55 including pharmaceutically acceptable salts, solvates and hydrates thereof. 5

40. A method of inhibiting the formation or accumulation of a visual cycle product, comprising contacting an opsin protein with a compound of claim 1. 10

41. The method of claim 40, wherein said compound reduces mislocalization of said opsin protein.

42. The method of claim 40, wherein said compound reduces hydration of said opsin protein. 15

43. The method of claim 40, wherein said compound binds to said opsin protein by hydrogen bonding.

44. The method of claim 40, wherein said opsin protein is present in a 20 cell.

45. The method of claim 44, wherein said cell is a cone cell or rod cell.

46. The method of claim 44, wherein said cell is present in a 25 mammalian eye.

47. The method of claim 46, wherein said mammal is a human being.

48. The method of claim 40, wherein said visual cycle product is a toxic 30 visual cycle product.

49. The method of claim 40, wherein said visual cycle product is lipofuscin or N-retinylidene-N-retinylethanolamine (A2E). 197

50. A method of treating or preventing an ophthalmic condition in a subject at risk thereof, comprising administering to the subject an effective amount of a compound of claim 1. 5

51. A method of treating or preventing an ophthalmic condition in a subject at risk thereof, comprising administering to the subject an effective amount of a compound of claim 21 including pharmaceutically acceptable salts thereof. 10

52. A method of treating or preventing an ophthalmic condition in a subject at risk thereof, comprising administering to the subject an effective amount of a compound of claim 37. 15

53. A method of treating or preventing an ophthalmic condition in a subject at risk thereof, comprising administering to the subject an effective amount of a compound of claim 38.

54. A method of treating or preventing an ophthalmic condition in a 20 subject at risk thereof, comprising administering to the subject an effective amount of a compound of claim 39.

55. The method of claim 51, wherein said ophthalmic condition is an ocular protein mislocalization disorder. 25

56. The method of claim 51, wherein said an ophthalmic condition is selected from the group consisting of wet or dry age related macular degeneration (ARMD), retinitis pigmentosa (RP), a retinal or macular dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant 30 drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity, normal vision loss related aging and normal loss of night vision related to aging . 198

57. The method of claim 56, wherein said ophthalmic condition is ARMD. 5

58. The method of claim 56, wherein said ophthalmic condition is light toxicity.

59. The method of claim 56, wherein said ophthalmic condition is retinitis pigmentosa (RP). 10

60. The method of claim 59, wherein said RP due to aberrant opsin folding.

61. The method of claim 52, wherein said ophthalmic condition is an 15 ocular protein mislocalization disorder.

62. The method of claim 52, wherein said an ophthalmic condition is selected from the group consisting of wet or dry age related macular degeneration (ARMD), retinitis pigmentosa (RP), a retinal or macular 20 dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity, normal vision loss related aging and normal loss of night vision related to aging . 25

63. The method of claim 62, wherein said ophthalmic condition is ARMD.

64. The method of claim 62, wherein said ophthalmic condition is light 30 toxicity.

65. The method of claim 62, wherein said ophthalmic condition is retinitis pigmentosa (RP). 199

66. The method of claim 65, wherein said RP due to aberrant opsin folding. 5

67. The method of claim 53 or 54, wherein said ophthalmic condition is an ocular protein mislocalization disorder.

68. The method of claim 67, wherein said an ophthalmic condition is selected from the group consisting of wet or dry age related macular 10 degeneration (ARMD), retinitis pigmentosa (RP), a retinal or macular dystrophy, Stargardt's disease, Sorsby's dystrophy, autosomal dominant drusen, Best's dystrophy, peripherin mutation associate with macular dystrophy, dominant form of Stargart's disease, North Carolina macular dystrophy, light toxicity, normal vision loss related aging and normal loss of 15 night vision related to aging .

69. The method of claim 68, wherein said ophthalmic condition is ARMED. 20

70. The method of claim 68, wherein said ophthalmic condition is light toxicity.

71. The method of claim 70, wherein said ophthalmic condition is retinitis pigmentosa (RP). 25

72. A composition, comprising a therapeutically effective amount of a compound of claim 1 in a pharmaceutically acceptable carrier.

73. A composition, comprising a therapeutically effective amount of a 30 compound of claim 21 in a pharmaceutically acceptable carrier.

74. A composition, comprising a therapeutically effective amount of a compound of claim 37 in a pharmaceutically acceptable carrier. 200

75. A composition, comprising a therapeutically effective amount of a compound of claim 38 in a pharmaceutically acceptable carrier. 5

76. A composition, comprising a therapeutically effective amount of a compound of claim 39 in a pharmaceutically acceptable carrier. Bikam Pharmaceuticals, Inc. Patent Attorneys for the Applicant/Nominated Person SPRUSON & FERGUSON