HK1196123B

HK1196123B - Alkynyl phenyl derivative compounds for treating ophthalmic diseases and disorders

Info

Publication number: HK1196123B
Application number: HK14109298.4A
Authority: HK
Inventors: 伊恩．莱斯莉．斯科特; 弗拉基米尔．亚历山德罗维奇．库克萨; 马克．W．奥姆; 托马斯．利特尔; 安娜．加尔; 珍妮弗．盖奇; 洪峰
Original assignee: 奥克塞拉有限公司
Priority date: 2007-06-29
Filing date: 2014-09-15
Publication date: 2016-09-23

Abstract

Provided are alkynyl phenyl derivative compounds, pharmaceutical compositions thereof, and methods of treating ophthalmic diseases and disorders, such as age-related macular degeneration and Stargardt's Disease, using said compounds and compositions.

Description

Alkynyl phenyl derivative compounds for the treatment of ophthalmic diseases and disorders

The present application is a divisional application of an invention patent application entitled "alkynyl phenyl derivative compounds for treating ophthalmic diseases and disorders" (international application No.: PCT/US 2008/008169), having application date of 30/6/2008, having application number of 200880104445.7.

Cross referencing

This application claims priority to U.S. provisional application No. 60/947,321, filed on 29/6/2007, which is hereby incorporated by reference in its entirety.

Background

Millions of patients worldwide have suffered from neurodegenerative diseases such as glaucoma, macular degeneration and Alzheimer's disease. Since the quality of life is greatly reduced by these diseases, research and development of drugs in this field are very important.

There are tens of millions to fifteen million patients in the united states who are profoundly afflicted with macular degeneration and have become a leading cause of blindness in the elderly around the world. Age-related macular degeneration affects central vision and causes photoreceptor cell depletion in the central portion of the retina known as the macula. Macular degeneration can be divided into two types: dry type and wet type. The dry form is more common than the wet form, and approximately 90% of age-related macular degeneration (AMD) patients are diagnosed as dry. The wet and geographic atrophy (geographica control), which is the terminal phenotype of dry AMD, of the disease causes the most severe visual loss. It was previously thought that all patients with wet form AMD had dry form AMD for a long period of time. The exact cause of age-related macular degeneration has not been known to date. The dry form of AMD can result from the aging and thinning of macular tissue associated with the deposition of pigment in the macular retinal pigment epithelium. In wet AMD, new blood vessels grow under the retina, forming scar tissue, leading to bleeding and exudation of fluid. The retina situated above it is severely damaged, creating a "blind" zone in central vision.

For the majority of patients with dry form macular degeneration, there is no effective treatment to date. Since the dry form progresses to wet form macular degeneration, medical intervention to prevent or delay the progression of dry form AMD is beneficial to patients with dry form AMD, and may reduce the incidence of wet form AMD.

The visual deterioration perceived by the patient or the typical features found by ophthalmologists in routine ophthalmic examinations may be the first signs of age-related macular degeneration. The formation of "drusen" or membrane debris under the retinal pigment epithelium of the macula is often the first physiological sign of AMD formation. Late symptoms include visual distortion of straight lines and, in some severe cases, dark, blurry or absent vision in central vision; and color vision changes may occur.

Different forms of genetically related macular degeneration may also occur in younger patients. The factors that cause the disease are genetic, nutritional, traumatic, infectious or other ecological factors.

Glaucoma is a generic term used to describe a series of diseases (usually asymptomatic) that cause a slowly progressive loss of visual field. The absence of symptoms may result in delayed diagnosis of glaucoma until the end of the disease. The prevalence of glaucoma in the united states is estimated at two hundred and twenty thousand, where blindness causes in about 120,000 cases can be attributed to this. This disease is particularly prevalent in japan, with approximately four million reported cases. In many parts of the world, people cannot receive treatment as conveniently as in the united states and japan, and thus glaucoma is a leading cause of blindness in the world worldwide. Even if patients with glaucoma are blind, their vision is often severely impaired.

The gradual loss of peripheral visual field in glaucoma is caused by the death of ganglion cells in the retina. Ganglion cells are specialized projection neurons that connect the eye and brain. Glaucoma is often accompanied by elevated intraocular pressure. Current treatments include the use of intraocular pressure-lowering drugs; however, current therapies for lowering intraocular pressure are often insufficient to completely inhibit the progression of the condition. Ganglion cells are considered to be pressure sensitive and may undergo permanent degeneration prior to lowering intraocular pressure. There are an increasing number of cases of glaucoma with normal eye pressure, in which ganglion cells are degenerated without an increase in intraocular pressure being observed. Current glaucoma medications only treat intraocular pressure and are not effective in preventing degeneration or reverse degeneration of ganglion cells.

Recent reports mention that glaucoma is a neurodegenerative disease, similar to alzheimer's disease and parkinson's disease of the brain, in addition to affecting especially retinal neurons. Retinal neurons of the eye originate from the internncephalon neurons of the brain. While retinal neurons are generally mistakenly considered not to be part of the brain, retinal cells are a key component of the central nervous system, which transmit signals from photoreceptor cells.

Alzheimer's Disease (AD) is the most common type of senile dementia. Dementia is a brain disorder that seriously affects a person's ability to perform daily activities. Four million patients with alzheimer's disease exist in the united states alone. The disease is characterized by loss of nerve cells in areas of the brain that are critical for memory and other mental functions. Currently commercially available drugs are capable of alleviating the symptoms of AD over a period of time, but there are no drugs that treat or completely inhibit progressive decline in mental function. Recent studies have shown that glial cells bearing (support) neurons or nerve cells may be deficient in patients with AD, but the cause of AD is not known to date. Individuals with AD appear to have a higher incidence of glaucoma and age-related macular degeneration, suggesting that similar pathogenesis may exist for these neurodegenerative diseases of the eye and brain (see Giasson, et al, freedric, biol. med.32:1264-75 (2002); johnonetal, proc.natl.acad.sci.usa99:11830-35 (2002); dentchevel. et al, mol.vis.9:184-90 (2003)).

Neuronal cell death explains the pathology of these diseases. Unfortunately, compositions and methods have been found to be scarce for improving the survival of retinal nerve cells, particularly photoreceptor cells. Accordingly, there is a need to identify and develop compositions that can be used to treat and prevent a range of retinal diseases and disorders in which neuronal cell death is a major or accessory factor in the pathogenesis.

In vertebrate photoreceptors, the emission of a photon causes isomerization of the 11-cis-retinylidene chromophore to all-trans-retinylidene and uncoupling from the visual opsin receptor. This photoisomerization causes a conformational change in the opsin protein which in turn causes a biological chain reaction called light transduction (filipekey., annu. rev. physiol.65:851-79 (2003)). Regeneration of the retinoid requires the re-conversion of the chromophore to the 11-cis-conformation during a process collectively known as the retinoid (visual) cycle (see, e.g., McBeeetal, prog.Retin. EyeRes.20:469-52 (2001)). First, the chromophore is released from the opsin and reduced in the photoreceptor by retinol dehydrogenase. The product, all-trans retinol, is entrapped in the vicinity of retinal pigment epithelial cells (RPEs) in the form of insoluble fatty acid esters in the subcellular structures of known retinal bodies (retinosomes) (imperial, j.cellbiol.164:373-87 (2004)).

In Skatagte's disease (Allikmettotal, Nat. Genet.15:236-46(1997)), with ABCR transporters acting as flippasesIn an allo-related disease, accumulation of all-trans retinal can cause the formation of lipofuscin pigment (A2E), which is toxic to retinal pigment epithelium and leads to progressive retinal degeneration, thus resulting in loss of vision (Mataeal, Proc. Natl. Acad. Sci. USA97:7154-59 (2000); Wengetal, Cell98:13-23 (1999)). The use of retinol dehydrogenase 13-cis-RA (isotretinoin, Accutane) has been considered Roche) as a therapy that may prevent and slow the formation of A2E and may have a protective effect in maintaining normal vision (radicetal, proc.natl.acad.sci.usa100:4742-47 (2003)). 13-cis-RA has been used to slow the synthesis of 11-cis-retinal by inhibiting 11-cis-RDH (Lawet., biochem. Biophys. Res. Commun.161:825-9(1989)), but its use also causes severe night blindness. Others have proposed that 13-cis-RA act to prevent regeneration of chromophores by binding to RPE65, a protein essential in the isomerization process of the eye (Gollapallial, Proc. Natl. Acad. Sci. USA101:10030-35 (2004)). Gollapalli et al reported that 13-cis-RA inhibited the formation of A2E, and suggested that this treatment could inhibit lipofuscin accumulation, thereby delaying the onset of loss of vision in Stargardt's disease or age-related macular degeneration, both of which are associated with lipofuscin accumulation associated with retinal pigment. However, inhibition of retinoid circulation and formation of uncoordinated opsins may lead to more serious outcomes and worsen patient prognosis (see, e.g., VanHooseltal, J.biol.chem.277:19173-82 (2002); Woodreffetal, nat. Gene.35: 158-164 (2003)). The lack of formation of the chromophore can lead to progressive retinal degeneration and produce a phenotype similar to Leber Congenital Amaurosis (LCA), a very rare genetic disease that affects children immediately after birth.

Disclosure of Invention

There is a need in the art for therapies for the treatment of recessive (hereditary) macular dystrophy and age-related macular degeneration (AMD), without causing other unwanted side effects, such as progressive retinal degeneration, LCA-like diseases, night blindness, or systemic vitamin a deficiency. There is also a need in the art to develop effective therapies for other ophthalmic diseases and disorders that are detrimental to the retina.

The present invention relates to alkynylphenyl derivative compounds that are inhibitors of the isomerization step of the vitamin a class cycle and are useful in the treatment of ophthalmic diseases and disorders. The invention also provides pharmaceutical compositions comprising the alkynylphenyl derivative compounds and methods of using these compounds to treat various ophthalmic disorders.

In one embodiment is a compound having the structure of formula (a):

the compound is a tautomer or a mixture of tautomers, or a pharmaceutically acceptable salt, hydrate, solvate, N-oxide, stereoisomer, geometric isomer, or prodrug thereof, wherein:

m is 0, 1, 2 or 3;

z is a bond, -C (R)¹)(R²)-、-X-C(R²¹)(R²²)-、-C(R²³)(R²⁴)-C(R¹)(R²) -or-C (R)²³)(R²⁴)-C(R²⁵)(R²⁶)-C(R¹)(R²)-、-X-C(R²¹)(R²²)-C(R¹)(R²)-、-C(R³²)(R³³)-X-C(R²¹)(R²²)-；

X is-O-, -S (= O)₂-、-N(R³¹)-、-C(=O)-、-C(=CH₂)-、-C(=N-NR³⁵) -OR-C (= N-OR) ³⁵)-；

Y is a bond, -C (R)²⁷)(R²⁸) -or-C (R)²⁷)(R²⁸)-C(R²⁹)(R³⁰)-；

R¹And R²Each independently selected from hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶or-NR⁷R⁸Performing the following steps; or R¹And R²Together form an oxo group;

R²¹、R²²、R³²and R³³Each independently selected from hydrogen and C₁-C₅In alkyl or fluoroalkyl;

R²³and R²⁴Each independently selected from hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Performing the following steps; or R²³And R²⁴Together form an oxo group; or optionally, R²³With adjacent R¹Together form a direct bond to provide a double bond; or optionally, R²³With adjacent R¹Together form a direct bond, and R²⁴With adjacent R²Together form a direct bond to provide a triple bond;

R²⁵and R²⁶Each independently selected from hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶or-NR⁷R⁸Performing the following steps; or R²⁵And R²⁶Together form an oxo group;

R³and R⁴Each independently selected from hydrogen, alkyl, heteroalkyl, alkenyl, fluoroalkyl, aryl, heteroaryl, carbocyclyl, or C-linked heterocyclyl; or R³And R⁴Together with the carbon atom to which they are attached form a carbocyclic or heterocyclic group; or R³And R⁴Together form an imino group;

R⁵is alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl, carbocyclyl, heteroaryl, or heterocyclyl;

R⁶each of which is the same or different and each is independently hydrogen or C ₁-C₅An alkyl group;

each R is⁷And each R⁸Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl, heteroalkyl, heterocycloalkyl, aryl, heteroaryl, -C (= O) R⁹、SO₂R⁹、CO₂R⁹、SO₂NH₂、SO₂NHR⁹Or SO₂N(R⁹)₂(ii) a Or R⁷And R⁸And together with the nitrogen atom to which they are attached form an N-heterocyclyl;

R⁹each being the same or different and each independently being an alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl group;

R¹²and R¹³Each of which is the same or different and each is independently hydrogen, alkyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -C (= O) R⁹、SO₂R⁹、CO₂R⁹、SO₂NH₂、SO₂NHR⁹Or SO₂N(R⁹)₂(ii) a Or R¹²And R¹³And together with the nitrogen atom to which they are attached form an N-heterocyclyl; and

R¹⁴each being the same OR different and each being independently alkyl, halogen, fluoroalkyl OR-OR⁶；

R²⁷、R²⁸、R²⁹And R³¹Each being the same OR different and each being independently hydrogen, alkyl OR-OR⁶(ii) a And

R³⁰and R³⁵Each independently is hydrogen or C₁-C₅An alkyl group.

In another embodiment are compounds of formula (A) wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²)-。

In yet another embodiment are compounds of formula (A) wherein R⁵Is an aryl group. In another embodiment are compounds of formula (A) wherein R⁵Is an unsaturated carbocyclic group. In yet another embodiment are compounds of formula (A) wherein R ⁵Is a bicyclic carbocyclic group. In another embodiment compounds of formula (A) wherein R⁵Is norbornyl (norbonyl).

In another embodiment are compounds of formula (A) wherein R⁵Is phenyl, Y is a bond, and the compound has the structure of formula (B):

m is 0, 1, 2 or 3;

n is 0, 1, 2, 3, 4 or 5;

R³and R⁴Each is independently selected from hydrogen and alkyl;

R⁶each of which is the same or different and each is independently hydrogen or C₁-C₅An alkyl group;

each R is⁷And each R⁸Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl or-C (= O) R⁹(ii) a Or R⁷And R⁸And together with the nitrogen atom to which they are attached form an N-heterocyclic ringA group;

R²³and R ²⁴Each of which is the same or different and each is independently hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Or a carbocyclic group; or R²³And R²⁴Together form an oxo group;

R¹²and R¹³Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R⁹(ii) a Or R¹²And R¹³And together with the nitrogen atom to which they are attached form an N-heterocyclyl;

R¹⁴each being the same OR different and each being independently alkyl, halogen, fluoroalkyl OR-OR⁶(ii) a And

R¹⁵each being the same OR different and each being independently alkyl, -OR⁶Alkenyl, alkynyl, halogen, fluoroalkyl, aryl, or aralkyl;

in another embodiment are compounds of formula (B) wherein R¹²And R¹³Each is hydrogen.

In yet another embodiment are compounds of formula (B) wherein R¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; wherein R is²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each independently hydrogen or alkyl.

In another embodiment are compounds of formula (B) wherein R¹And R²Each independentlyIs hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R ²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each is hydrogen.

In yet another embodiment are compounds of formula (B) wherein m is 0; n is 0, 1 or 2; and R¹⁵Each independently is alkyl, -OR⁶Or an aryl group.

In another embodiment, the compound is selected from: 3- (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propan-1-amine; 3- (3- ((2-methoxyphenyl) ethynyl) phenyl) propan-1-amine; 3- (3- (phenylethynyl) phenyl) propan-1-amine; 3-amino-1- (3- (biphenyl-3-ylethynyl) phenyl) propan-1-ol; and 3-amino-1- (3- ((2-methoxyphenyl) ethynyl) phenyl) propan-1-ol.

In another embodiment are compounds of formula (A) wherein R⁵Is 1-naphthyl or 2-naphthyl. In yet another embodiment are those wherein R¹、R²、R³、R⁴、R²³And R²⁴Each hydrogen. In another embodiment are compounds of formula (a) wherein m is 0.

In another embodiment is the compound 3- (3- (naphthalen-2-ylethynyl) phenyl) propan-1-amine.

In another embodiment are those in which R⁵Is C (R)¹⁶)(R¹⁷)(R¹⁸) A compound of formula (a) wherein Y is a bond, and the compound has a structure of formula (C):

m is 0, 1, 2 or 3;

R¹and R²Each of which is the same or different and each is independently hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Or a carbocyclic group; or R¹And R²Together form an oxo group;

R³and R⁴Each is the same or different and each is independently hydrogen or alkyl;

each R is⁶Are the same or different and are each independently hydrogen or C₁-C₅An alkyl group;

each R is⁷And each R⁸Are the same or different and are each independently hydrogen, alkyl, carbocyclyl or-C (= O) R⁹(ii) a Or R⁷And R⁸Together with the N atom to which they are attached form an N-heterocyclyl;

each R is⁹The same or different and are independently alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R²³and R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸(ii) a Or R²³And R²⁴Together form an oxo group;

R¹²and R¹³Are the same or different and are independently hydrogen, alkyl or-C (= O) R⁹(ii) a Or R¹²And R¹³Together with the N atom to which they are attached form an N-heterocyclyl;

each R is¹⁴The same OR different and independently is alkyl, halogen, fluoroalkyl OR-OR⁶(ii) a And

R¹⁶、R¹⁷and R¹⁸Each being the same or different and each being independentThe site is hydrogen, alkyl, -OR⁶Carbocyclyl or aryl.

In another embodiment are those in which R¹²And R¹³Compounds of the general formula (C) each being hydrogen. In another embodiment are compounds of formula (C) wherein R¹And R²Each independently of the others is hydrogen, halogen, C_1-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each independently hydrogen or alkyl. In another embodiment are compounds of formula (C) wherein m is 0, and R is¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each is hydrogen. In another embodiment are compounds of formula (C) wherein R¹⁶、R¹⁷And R¹⁸Each independently is hydrogen, alkyl, carbocyclyl or aryl.

In another embodiment of the invention are compounds selected from the group consisting of: 4- (3- (3-aminopropyl) phenyl) but-3-yn-1-ol; 5- (3- (3-aminopropyl) phenyl) pent-4-yn-2-ol; 2- (3- (3-cyclopentylprop-1-ynyl) phenoxy) ethylamine; 3- (3- (3, 3-dimethylbut-1-ynyl) phenyl) propan-1-amine; 3- (3- (3-phenylprop-1-ynyl) phenyl) propan-1-amine; 3- (3- (pent-1-ynylphenyl) prop-1-amine, 3- (3- (hex-1-ynylphenyl) prop-1-amine, 3-amino-1- (3- (3-cyclopentylprop-1-ynylphenyl) phenyl) prop-1-ol, 3-amino-1- (3- (3-phenylprop-1-ynylphenyl) prop-1-ol, 6- (3-amino-1-hydroxypropyl) phenyl) hex-5-yn-1-ol, 4- (3- (3-amino-1-hydroxypropyl) phenyl) but-3-yn-1-ol, 3-amino-1- (3- (hept-1- Alkynyl) phenyl) propan-1-ol; 3- (3- (4-phenylbut-1-ynyl) phenyl) propan-1-amine; 3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol; 3- (3- (5-methoxypent-1-ynyl) phenyl) propan-1-amine; 3-amino-1- (3- (4-phenylbut-1-ynyl) phenyl) propan-1-ol; 6- (3- (3-aminopropyl) phenyl) hex-5-yn-1-ol; and 3- (3- (6-methoxyhex-1-ynyl) phenyl) prop-1-amine.

In another embodiment are compounds of formula (C) wherein R¹⁶is–OR⁶Wherein R is⁶Is hydrogen or C₁-C₅Alkyl, and R¹⁷And R¹⁸Each independently hydrogen, alkyl or aryl.

In another embodiment are compounds selected from the group consisting of:

1- (3- (3-aminopropyl) phenyl) -3-ethylpent-1-yn-3-ol; 4- ((3- (3-aminopropyl) phenyl) ethynyl) heptan-4-ol; 5- ((3- (3-aminopropyl) phenyl) ethynyl) nonan-5-ol; 3- (3- (3-methoxy-3-propylhex-1-ynyl) phenyl) propan-1-amine; 1- (3- (3-aminopropyl) phenyl) -3-methylhexan-1-yn-3-ol; 1- (3- (3-aminopropyl) phenyl) -3, 4-dimethylpent-1-yn-3-ol; 4- (3- (3-aminopropyl) phenyl) -2-methylbut-3-yn-2-ol; 1- (3- (3-aminopropyl) phenyl) hex-1-yn-3-ol; 1- (3- (3-aminopropyl) phenyl) -3, 4-dimethylhex-1-yn-3-ol; 3- (3- (3-methoxyprop-1-ynyl) phenyl) prop-1-amine; 3- (3- (3-aminopropyl) phenyl) prop-2-yn-1-ol; 1- (3- (3-aminopropyl) phenyl) -3-tert-butyl-4, 4-dimethylpent-1-yn-3-ol; (R) -1- (3- (3-aminopropyl) phenyl) oct-1-yn-3-ol; (R) -1- (3- (3-aminopropyl) phenyl) oct-1-yn-3-ol; (R) -3- (3- (3-aminopropyl) phenyl) -1-phenylprop-2-yn-1-ol; 4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) heptan-4-ol; 4- ((3- (3-amino-2, 2-dimethylpropyl) phenyl) ethynyl) hept-4-ol; 4- (3- (3-aminopropyl) phenyl) -2-phenylbut-3-yn-2-ol; 1- (3- (3-aminopropyl) phenyl) -4-methylpent-1-yn-3-ol; 1- (3- (3-aminopropyl) phenyl) -3,4, 4-trimethylpent-1-yn-3-ol; (R) -3- (3- (3-aminopropyl) phenyl) -1-phenylprop-2-yn-1-ol; 1- (3- (3-aminopropyl) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol; 4- ((3- (3-aminopropyl) phenyl) ethynyl) -2, 6-dimethylhept-4-ol; 1- (3- (3-amino-1-hydroxypropyl) phenyl) -3-ethylpent-1-yn-3-ol; 3- (3- (3-ethylpent-1-ynyl) phenyl) propan-1-amine; 3- (3- (3-propylhex-1-ynyl) phenyl) prop-1-amine; 3-amino-1- (3- (3-ethylpent-1-ynyl) phenyl) propan-1-ol; 3-amino-1- (3- (3-propylhex-1-ynyl) phenyl) propan-1-ol; 3-amino-1- (3- (3-ethylpent-1-ynyl) phenyl) -2-methylpropan-1-ol; 1- (3- (3-amino-1-hydroxy-2-methylpropyl) phenyl) -3-ethylpent-1-yn-3-ol; 1-amino-3- (3- (3-ethylpent-1-ynyl) phenyl) propan-2-ol; 1- (3- (3-amino-2-hydroxypropyl) phenyl) -3-ethylpent-1-yn-3-ol; 3-amino-2-methyl-1- (3- (3-propylhex-1-ynyl) phenyl) propan-1-ol; 4- ((3- (3-amino-1-hydroxy-2-methylpropyl) phenyl) ethynyl) hept-4-ol; 4- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) heptan-4-ol; 1-amino-3- (3- (3-propylhex-1-ynyl) phenyl) propan-2-ol.

In another embodiment are those wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²) -and R⁵A compound of the general formula (a) which is a carbocyclic group.

In another embodiment are those in which R⁵A compound of formula (a) which is cycloalkyl, Y being a bond, and which has the structure of formula (D):

m is 0, 1, 2 or 3;

p is 1, 2, 3, 4, 5 or 6;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

each R is⁷And each R⁸Each of which is the same or different and is independently hydrogen, alkyl, carbocyclyl, heteroalkyl, heterocycloalkyl, aryl, heteroaryl, or-C (= O) R⁹(ii) a Or R⁷And R⁸Together with the N atom to which they are attached form an N-heterocyclyl;

Each R is⁹The same or different and each is independently alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R²³and R²⁴Each of which is the same or different and each is independently hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Or a carbocyclic group; or R²³And R²⁴Together form an oxo group;

R¹²and R¹³Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R⁹(ii) a Or R¹²And R¹³Together with the N atom to which they are attached form an N-heterocyclyl;

each R is¹⁴Are the same OR different and are each independently alkyl, halogen, fluoroalkyl OR-OR⁶(ii) a And

each R is¹⁹Are the same OR different and are each independently alkyl, -OR⁶Halogen or fluoroalkyl.

In another embodiment are those in which R¹²And R¹³Compounds of the general formula (D) each being hydrogen. In another implementationIn the formula (D), wherein R is¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen or alkyl, wherein R⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each independently hydrogen or alkyl. In another embodiment are compounds of formula (D) wherein p is 3, and R is⁵Is a substituted or unsubstituted cyclopentyl group. In another embodiment are compounds of formula (D) wherein p is 4, and R is ⁵Is a substituted or unsubstituted cyclohexyl group. In another embodiment are compounds of formula (D) wherein p is 5, and R is⁵Is a substituted or unsubstituted cycloheptyl. In another embodiment are compounds of formula (D) wherein R¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each is hydrogen. In another embodiment are compounds of formula (D) wherein m is 0. In another embodiment are compounds of formula (D) wherein q is 0. In another embodiment are compounds of formula (D) wherein q is 1, 2, 3, 4, or 5, and each R is¹⁹Independently is alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group.

In another embodiment is a compound selected from the group consisting of:

3- (3- (cyclopentylethynyl) phenyl) propan-1-amine; 3- (3- (cyclohexylethynyl) phenyl) propan-1-amine; 3-amino-1- (3- (cyclopentylethynyl) phenyl) propan-1-ol; 3-amino-1- (3- (cyclohexylethynyl) phenyl) propan-1-ol; 1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclohexanol; 1- ((3- (3-aminopropyl) phenyl) ethynyl) -2,2,6, 6-tetramethylcyclohexanol; 1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclopentanol; 3- (3- (cycloheptylethynyl) phenyl) propan-1-amine; 3-amino-1- (3- (cycloheptylethynyl) phenyl) propan-1-ol; 1-amino-3- (3- (cycloheptylethynyl) phenyl) propan-2-ol; 1-amino-3- (3- (cyclohexylethynyl) phenyl) propan-2-ol; 1-amino-3- (3- (cyclopentylethynyl) phenyl) propan-2-ol; 1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cyclopentanol; 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclopentanol; 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclohexanol; 1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cycloheptanol; 1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cyclohexanol; 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cycloheptanol; and 1- ((3- (3-aminopropyl) phenyl) ethynyl) cycloheptanol.

In another embodiment are compounds of formula (D) wherein R¹²Is hydrogen, and R¹³is-C (= O) R⁹Wherein R is⁹Is an alkyl group. In another embodiment are compounds of formula (D) wherein R¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each is hydrogen. In another embodiment are compounds of formula (D) wherein m is 0. In another embodiment is the compound of claim 36, wherein q is 1, 2, 3, 4, or 5, and each R is¹⁹Independently is alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group. In another embodiment are compounds of formula (D) wherein p is 4, and R is⁵Is a substituted or unsubstituted cyclohexyl group.

In another embodiment is the compound N- (3-hydroxy-3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide.

In another embodiment are compounds of formula (A) wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²) -, wherein R⁵Is a heterocyclic group and Y is a bond. In another implementationIn which the heterocyclic radical may be-OR⁶Optionally substituted compound, wherein R⁶Is hydrogen or C₁-C₅An alkyl group. In another embodiment are those in which R ¹²And R¹³Each hydrogen.

In another embodiment are compounds of formula (A) wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²) -, wherein Y is a bond, R⁵Is a heterocyclic group, and the heterocyclic group may be substituted by-OR⁶Optionally substituted, wherein R⁶Is hydrogen or C₁-C₅Alkyl, and R¹²And R¹³Each is hydrogen, and wherein R¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each independently hydrogen or alkyl.

In another embodiment are compounds of formula (A) wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²) -, wherein Y is a bond, R⁵Is a heterocyclic group, and the heterocyclic group may be substituted by-OR⁶Optionally substituted, wherein R⁶Is hydrogen or C₁-C₅Alkyl, and R¹²And R¹³Each is hydrogen, and wherein R¹、R²、R³、R⁴、R²³And R²⁴Each is hydrogen. In another embodiment are compounds wherein m is 0.

In another embodiment the compound is selected from the following: 4- ((3- (3-aminopropyl) phenyl) ethynyl) tetrahydro-2H-thiopyran-4-ol; and 4- ((3- (3-aminopropyl) phenyl) ethynyl) tetrahydro-2H-pyran-4-ol.

In another embodiment are compounds of formula (A) wherein Z is-C (R) ²³)(R²⁴)-C(R¹)(R²) -, and R⁵Is heteroaryl, and Y is a bond. In another embodiment are those in which R¹²And R¹³Each hydrogen. In another embodiment are compounds wherein the substituent is a group wherein R is¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each independently hydrogen or alkyl. In another embodiment are those in which R¹、R²、R³、R⁴、R²³And R²⁴Each hydrogen. In another embodiment are compounds wherein m is 0.

In another embodiment is a compound selected from the group consisting of: 3- (3- (pyridin-2-ylethynyl) phenyl) propan-1-amine; 3- (3- (pyridin-3-ylethynyl) phenyl) propan-1-amine; 3- (3- (pyridin-4-ylethynyl) phenyl) propan-1-amine; 3- (3- (phenylthio-2-ylethynyl) phenyl) propan-1-amine; and 3- (3- (phenylthio-3-ylethynyl) phenyl) propan-1-amine.

In another embodiment are compounds of formula (A) wherein Z is-O-C (R)²¹)(R²²) -Y is a bond, and the compound has the structure of formula (E):

m is 0, 1, 2 or 3;

R²¹and R²²Each independently is hydrogen, C₁-C₅Alkyl or fluoroalkyl;

R⁵is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R⁹is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R¹²and R¹³Are the same or different and are independently hydrogen, alkyl or-C (= O) R⁹(ii) a Or R¹²And R¹³Together with the N atom to which they are attached form an N-heterocyclyl; and

each R is¹⁴Are the same OR different and are each independently alkyl, halogen, fluoroalkyl OR-OR⁶。

In another embodiment are compounds of formula (E) wherein R⁵Is an unsaturated carbocyclic group. In another embodiment are compounds of formula (E) wherein R⁵Is a bicyclic carbocyclic group. In another embodiment are compounds of formula (E) wherein R⁵Is norbornyl.

In another embodiment are compounds of formula (E) wherein R⁵is-C (R)¹⁶)(R¹⁷)(R¹⁸) Y is a bond, and the compound has the structure of formula (F):

m is 0, 1, 2 or 3;

R²¹and R²²Each independently is hydrogen, C₁-C₅Alkyl or fluoroalkyl;

R⁹is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

each R is¹⁴Are the same OR different and are each independently alkyl, halogen, fluoroalkyl OR-OR⁶(ii) a And R¹⁶、R¹⁷、R¹⁸Each being the same OR different and each being independently hydrogen, alkyl, -OR⁶Carbocyclyl or aryl.

In another embodiment are compounds of formula (F) wherein R¹²And R¹³Each is hydrogen. In another embodiment are compounds of formula (F) wherein R²¹、R²²、R³And R⁴Each independently is hydrogen or C₁-C₅An alkyl group. In another embodiment are compounds of formula (F) wherein m is 0. In another embodiment are compounds of formula (F) wherein R¹⁶、R¹⁷And R¹⁸Each independently hydrogen, alkyl OR-OR⁶Wherein each R is⁶Independently is hydrogen or C₁-C₅An alkyl group. In another embodimentIs a compound of the general formula (F) wherein R ¹⁶、R¹⁷And R¹⁸Each independently hydrogen, alkyl or aryl.

In another embodiment is a compound selected from the group consisting of: 4- ((3- (2-aminoethoxy) phenyl) ethynyl) heptan-4-ol; 1- (3- (2-aminoethoxy) phenyl) -3-ethylpent-1-yn-3-ol; 1- (3- (2-aminoethoxy) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol; 5- ((3- (2-aminoethoxy) phenyl) ethynyl) nonan-5-ol; 4- (3- (2-aminoethoxy) phenyl) -2-methylbut-3-yn-2-ol; 2- (3- (hept-1-ynyl) phenoxy) ethylamine; 4- (3- (2-aminoethoxy) phenyl) but-3-yn-1-ol; 2- (3- (3-phenylprop-1-ynyl) phenoxy) ethylamine; 2- (3- (4-methylpent-1-ynyl) phenoxy) ethylamine; 6- (3- (2-aminoethoxy) phenyl) hex-5-yn-1-ol; 2- (3- (3-ethylpent-1-ynyl) phenoxy) prop-1-amine; 2- (3- (3-propylhex-1-ynyl) phenoxy) prop-1-amine; 1- (3- (2-aminoethoxy) phenyl) -3-ethylpent-1-yn-3-ol; 4- ((3- (1-aminopropyl-2-oxy) phenyl) ethynyl) hept-4-ol; 2- (3- (3-ethylpent-1-ynyl) phenoxy) ethylamine; and 2- (3- (3-propylhex-1-ynyl) phenoxy) ethylamine.

In another embodiment are compounds of formula (E) wherein R⁵Is a carbocyclyl group. In another embodiment are compounds of formula (E) wherein R ⁵Is cycloalkyl, Y is a bond, and the compound has the structure of formula (G):

m is 0, 1, 2 or 3;

p is 1, 2, 3, 4 or 5;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

R²¹and R²²Each of which is the same or different and each is independently hydrogen, C₁-C₅Alkyl or fluoroalkyl;

R⁹is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

In another embodiment are compounds of formula (G) wherein R ¹²And R¹³Each is hydrogen. In another embodiment are compounds of formula (G) wherein m is 0. In another embodiment are compounds of formula (G) wherein R²¹And R²²Each independently is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each independently hydrogen or alkyl. In another embodiment are compounds of formula (G) wherein R²¹、R²²、R³And R⁴Each is hydrogen or C₁-C₅An alkyl group. In another embodiment are compounds of formula (G) wherein q is 0 or 1, and each R is¹⁹Independently is alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group.

In another embodiment is a compound selected from the group consisting of: 1- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclopentanol; 1- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclohexanol; 2- (3- (cyclohexylethynyl) phenoxy) ethylamine; 1- ((3- (1-aminopropyl-2-yloxy) phenyl) ethynyl) cyclopentanol; 1- ((3- (1-aminopropyl-2-yloxy) phenyl) ethynyl) cyclohexanol; 1- ((3- (1-aminopropyl-2-yloxy) phenyl) ethynyl) cycloheptanol; 2- (3- (cycloheptylethynyl) phenoxy) ethylamine; 2- (3- (cycloheptylethynyl) phenoxy) propan-1-amine; 2- (3- (cyclohexylethynyl) phenoxy) propan-1-amine; 2- (3- (cyclopentylethynyl) phenoxy) propan-1-amine; 2- (3- (cyclopentylethynyl) phenoxy) -ethylamine; and 1- ((3- (2-aminoethoxy) phenyl) ethynyl) -cycloheptanol.

In another embodiment are compounds of formula (E) wherein R⁵Is a heterocyclic group. In another embodiment are compounds of formula (E) wherein m is 0, and R is¹²And R¹³Each is hydrogen. In another embodiment are compounds of formula (E) wherein R²¹、R²²、R³And R⁴Each independently is hydrogen or C₁-C₅An alkyl group.

In another embodiment is the compound 2- (3- (pyridin-3-ylethynyl) phenoxy) ethylamine.

In another embodiment are compounds of formula (E) wherein R⁵Is an aryl group.

In another embodiment is the compound 2- (3- (phenylethynyl) phenoxy) ethylamine.

In another embodiment is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of any one of formulae (a) to (G).

In another embodiment, the IC50 for inhibiting 11-cis-retinol production is a compound of less than about 1 μ M (as measured in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week. In a specific embodiment, the compound inhibits 11-cis-retinol production with an IC50 of less than about 100nM (as measured in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week. In yet another embodiment, the compound inhibits 11-cis-retinol production with an IC50 of less than about 10nM (as measured in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week, one month, two months, four months, six months, eight months, ten months, one year, two years, five years, or more.

In another embodiment is a non-retinoid compound that inhibits the isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value of less than 1mg/kg when administered to an individual. In another embodiment is a non-retinoid compound, wherein the ED50 value is measured after about 2 hours or more after administering a single dose of the compound to the patient. In yet another embodiment, the compound is a compound having an amine with an alkynylphenyl attached. In yet another embodiment, the compound is a non-retinoid compound.

In another embodiment is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ M (as determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week. In yet another embodiment is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a non-retinoid compound that inhibits an isomerase reaction that results in production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value of less than 1mg/kg when administered to an individual.

In another embodiment, the present invention provides a method of modulating the amount of a chromophore in the retinoid circulation, the method comprising administering to a subject a compound disclosed herein, the compound comprising any one of formulae (a) - (G) and their respective substructures. In yet another embodiment, the method results in a reduction in lipofuscin pigment accumulation in the eye of the subject. In yet another embodiment, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In yet another embodiment is a method of treating an ophthalmic disease or disorder in a subject comprising administering to the subject a compound or pharmaceutical composition disclosed herein. In yet another embodiment, the ophthalmic disease or disorder is age-related macular degeneration or stargardt disease. In yet another embodiment, the method results in a reduction in lipofuscin pigment accumulation in the eye of the subject. In yet another embodiment, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In yet another embodiment, the ophthalmic disease or disorder is selected from retinal detachment, hemorrhagic retinopathy, retinitis pigmentosa, cone-rod dystrophy, Sorsby fundus dystrophy, optic neuropathy, inflammatory retinopathy, diabetic maculopapulopathy, retinal vessel occlusion, retinopathy of prematurity or ischemia-reperfusion related retinal damage, proliferative vitreoretinopathy, retinal dystrophy, hereditary optic neuropathy, Sorsby fundus dystrophy, uveitis, retinal damage, retinal disorders associated with alzheimer's disease, retinal disorders associated with multiple sclerosis, retinal disorders associated with parkinson's disease, retinal disorders associated with viral infection, retinal disorders associated with excessive light stimulation, myopia, and retinal disorders associated with AIDS.

In another embodiment is a method of inhibiting dark adaptation of rod photoreceptor cells of a retina, the method comprising contacting the retina with a compound disclosed herein, the compound comprising any one of formulae (a) - (G) and their respective substructures.

In another embodiment is a method of inhibiting rhodopsin regeneration in rod photoreceptor cells of the retina, the method comprising contacting the retina with: any one of compounds of general formulae (a) to (G) and their respective substructures; a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ Μ (determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week; alternatively, a non-retinoid compound that inhibits an isomerase reaction that results in production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value below 1mg/kg when administered to an individual.

In another embodiment is a method of reducing ischemia in an eye of an individual comprising administering to the individual a pharmaceutical composition comprising: any one of compounds of general formulae (a) to (G) and their respective substructures; a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ Μ (determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week; or a non-retinoid compound that inhibits an isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value below 1mg/kg when administered to an individual. In yet another embodiment, the pharmaceutical composition is administered under conditions and for a time sufficient to inhibit dark adaptation of rod photoreceptor cells, thereby reducing ischemia in the eye.

In yet another embodiment is a method of inhibiting the formation of new blood vessels in the retina of an eye of an individual comprising administering to the individual a pharmaceutical composition of any one of compounds of formulae (a) - (G) and their respective substructures. In a specific embodiment, the pharmaceutical composition is administered under conditions and for a time sufficient to inhibit dark adaptation of rod photoreceptor cells, thereby inhibiting neovascularization in the retina.

In yet another embodiment is a method of inhibiting retinal cell degeneration in the retina comprising contacting the retina with: a compound of the general formula (A); a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ Μ (determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week; or a non-retinoid compound that inhibits an isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE, and the compound has an ED50 value below 1mg/kg when administered to an individual. In yet another embodiment, the pharmaceutical composition is administered under conditions and for a time sufficient to inhibit dark adaptation of rod photoreceptor cells, thereby reducing ischemia in the eye. In a specific embodiment is a method wherein the retinal cell is a retinal nerve cell. In one embodiment, the retinal nerve cell is a photoreceptor cell.

In yet another embodiment, there is provided a method of treating an ophthalmic disease or disorder in a subject comprising administering to the subject a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound having any of the structures of formulae (a) - (G) described above and herein. In one embodiment, the ophthalmic disease or disorder is a retinal disease or disorder. In a specific embodiment, the retinal disease or disorder is age-related macular degeneration or stargardt's macular dystrophy. In yet another embodiment, the ophthalmic disease or disorder is selected from the group consisting of retinal detachment, hemorrhagic retinopathy, retinitis pigmentosa, optic neuropathy, inflammatory retinopathy, proliferative vitreoretinopathy, retinal dystrophy, hereditary optic neuropathy, Sorsby fundus dystrophy, uveitis, retinal damage, retinal disorders associated with alzheimer's disease, retinal disorders associated with multiple sclerosis, retinal disorders associated with parkinson's disease, retinal disorders associated with viral infection, retinal disorders associated with excessive light stimulation, and retinal disorders associated with AIDS. In yet another embodiment, the ophthalmic condition or disorder is selected from the group consisting of diabetic retinopathy, diabetic maculopapulosis, retinal vessel occlusion, retinopathy of prematurity, or ischemia reperfusion-related retinal injury.

The present invention further provides a method of reducing lipofuscin pigment accumulation in the retina of an individual comprising administering to the individual a pharmaceutical composition described herein. In one embodiment, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In another embodiment, a method of inhibiting a trans-cis isomerase of at least one visual cycle in a cell is provided, wherein the method comprises contacting the cell with a compound having any one of structures (a) - (G) as described herein, thereby inhibiting the trans-cis isomerase of at least one visual cycle. In one embodiment, the cell is a Retinal Pigment Epithelial (RPE) cell.

In another embodiment, there is also provided a method of inhibiting a trans-cis isomerase enzyme of at least one visual cycle in a subject, the method comprising administering to the subject a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound having any of the structures of formulae (a) - (G) as described herein. In one embodiment, the individual is a human or non-human animal.

In particular embodiments of the methods described above and herein, the accumulation of lipofuscin pigment in the eye of the subject is inhibited, and in a particular embodiment, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E). In another certain embodiment, degeneration of retinal cells is inhibited. In a specific embodiment, the retinal cell is a retinal nerve cell, wherein the retinal nerve cell is a photoreceptor cell, an amacrine cell, a horizontal cell, a ganglion cell, or a bipolar cell. In another specific embodiment, the retinal cell is a Retinal Pigment Epithelium (RPE) cell.

In addition, in one embodiment, compounds having the structure of formula (I) are provided.

The compound is a tautomer or a mixture of tautomers, or a pharmaceutically acceptable salt, hydrate, solvate, N-oxide, or prodrug thereof, wherein:

m is 0, 1, 2 or 3;

R₁and R₂Each being the same OR different and each being independently hydrogen, halogen, alkyl, fluoroalkyl, -OR₆、-NR₇R₈Or a carbocyclic group; or

R₁And R₂Forming an oxo group;

R₃and R₄Each is the same or different and each is independently hydrogen or alkyl;

R₅is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₆is hydrogen or alkyl;

R₇and R₈Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl or-C (= O) R₉(ii) a Or R₇And R₈And together with the nitrogen atom to which they are attached form an N-heterocyclyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

x is-C (R)₁₀)(R₁₁) -or-O-;

R₁₀and R₁₁Each being the same OR different and each being independently hydrogen, halogen, alkyl, fluoroalkyl, -OR₆、-NR₇R₈Or a carbocyclic group; or

R₁₀And R₁₁Forming an oxo group;

R₁₂and R₁₃Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R ₉(ii) a Or R₁₂And R₁₃Together with the nitrogen atom to which they are attached form an N-heterocyclyl; and

each R is₁₄The same OR different and independently is alkyl, halogen, fluoroalkyl OR-OR₆。

The invention also provides compounds having the general formula (II), (IIa), (IIb), (IIc), (III), (IIIa) and (IIIb).

Wherein m, n, p, q, R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈And R₁₉As defined above and herein (see the detailed description section).

Another embodiment provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound having the structure of formula (I):

wherein R is₁、R₂、R₃、R₄、R₅、R₁₂、R₁₃、R₁₄And X is as defined herein.

The invention also provides compounds comprising structures having any of the general formulas (II), (IIa), (IIb), (IIc), (III), (IIIa), and (IIIb):

wherein m, n, p, q, R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈And R₁₉As described above and defined herein (see the detailed description section).

In another embodiment is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound disclosed herein, including, but not limited to, compounds of formulae (a) - (G) and (I) - (III) and their respective substructures.

In yet another embodiment, the IC50 for inhibiting 11-cis-retinol production is a compound of about 1 μ M or less (as determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week. In a specific embodiment, the compound inhibits 11-cis-retinol production with an IC50 of less than about 100nM (as measured in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week. In yet another embodiment, the compound inhibits 11-cis-retinol production with an IC50 of less than about 10nM (as measured in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week, one month, two months, four months, six months, eight months, ten months, one year, two years, five years, or more.

In another embodiment is a non-retinoid compound that inhibits the isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value of less than 1mg/kg when administered to an individual. In another embodiment is a non-retinoid compound, wherein the ED50 value is measured after about 2 hours or more after administering a single dose of the compound to the patient. In yet another embodiment, the compound is an amine compound having an alkynylphenyl group attached. In yet another embodiment, the compound is a non-retinoid compound.

In another embodiment, the present invention provides a method of modulating the amount of a chromophore in the retinoid circulation, the method comprising administering to a subject a compound disclosed herein, the compound comprising any one of formulae (a) - (G) and (I) - (III) and their respective substructures. In yet another embodiment, the method results in a reduction in lipofuscin pigment accumulation in the eye of the subject. In yet another embodiment, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In yet another embodiment is a method of treating an ophthalmic disease or disorder in a subject comprising administering to the subject a compound or pharmaceutical composition disclosed herein. In yet another embodiment, the ophthalmic disease or disorder is age-related macular degeneration or stargardt's macular dystrophy. In yet another embodiment, the method results in a reduction in lipofuscin pigment accumulation in the eye of the subject. In yet another embodiment, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In another embodiment is a method of inhibiting dark adaptation of rod photoreceptor cells of the retina, the method comprising contacting the retina with a compound disclosed herein, the compound comprising any one of formulae (a) - (G) and (I) - (III) and their respective substructures.

In another embodiment is a method of inhibiting rhodopsin regeneration in rod photoreceptor cells of the retina, the method comprising contacting the retina with: any one of the compounds of the general formulae (A) - (G) and (I) - (III) and their respective substructures; a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ Μ (determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week; alternatively, a non-retinoid compound that inhibits an isomerase reaction that results in production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value below 1mg/kg when administered to an individual.

In another embodiment is a method of reducing ischemia in an eye of an individual comprising administering to the individual a pharmaceutical composition comprising: any one of the compounds of the general formulae (A) to (G) and (I) to (III) and their respective substructures; a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ Μ (determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week; or a non-retinoid compound that inhibits an isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value below 1mg/kg when administered to an individual. In yet another embodiment, the pharmaceutical composition is administered under conditions and for a time sufficient to inhibit dark adaptation of rod photoreceptor cells, thereby reducing ischemia in the eye.

In yet another embodiment is a method of inhibiting neovascularization in the retina of an eye of an individual comprising administering to the individual a pharmaceutical composition of any one of compounds of formulae (a) - (G) and (I) - (III) and their respective substructures. In a specific embodiment, the pharmaceutical composition is administered under conditions and for a time sufficient to inhibit dark adaptation of rod photoreceptor cells, thereby inhibiting neovascularization in the retina.

In yet another embodiment is a method of inhibiting retinal cell degeneration in the retina comprising contacting the retina with: a compound of the general formula (A); a compound that inhibits the production of 11-cis-retinol with an IC50 of less than about 1 μ Μ (determined in vitro using an extract of RPE65 and LRAT expressing cells), wherein the extract further comprises CRALBP, wherein the compound is stable in solution at room temperature for at least about one week; or a non-retinoid compound that inhibits an isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value below 1mg/kg when administered to an individual. In yet another embodiment, the pharmaceutical composition is administered under conditions and for a time sufficient to inhibit dark adaptation of rod photoreceptor cells, thereby reducing ischemia in the eye. In a specific embodiment is a method wherein the retinal cell is a retinal nerve cell. In one embodiment, the retinal nerve cell is a photoreceptor cell.

In yet another embodiment, there is provided a method of treating an ophthalmic disease or disorder in a subject, comprising administering to the subject a pharmaceutical composition comprising a pharmaceutically acceptable carrier and any one of the compounds having the general formulae (a) - (G) and (I) - (III) and their respective substructures described herein. In one embodiment, the ophthalmic disease or disorder is a retinal disease or disorder. In a specific embodiment, the retinal disease or disorder is age-related macular degeneration or stargardt's macular dystrophy. In yet another embodiment, the ophthalmic disease or disorder is selected from the group consisting of retinal detachment, hemorrhagic retinopathy, retinitis pigmentosa, optic neuropathy, inflammatory retinopathy, proliferative vitreoretinopathy, retinal dystrophy, hereditary optic neuropathy, Sorsby fundus dystrophy, uveitis, retinal damage, retinal disorders associated with alzheimer's disease, retinal disorders associated with multiple sclerosis, retinal disorders associated with parkinson's disease, retinal disorders associated with viral infection, retinal disorders associated with excessive light stimulation, and retinal disorders associated with AIDS. In yet another embodiment, the ophthalmic condition or disorder is selected from the group consisting of diabetic retinopathy, diabetic maculopapulosis, retinal vessel occlusion, retinopathy of prematurity, or ischemia reperfusion-related retinal injury.

In another embodiment, a method of inhibiting a trans-cis isomerase of at least one visual cycle in a cell is provided, wherein the method comprises contacting the cell with a compound having any one of formulae (a) - (G) and (I) - (III) as described herein, thereby inhibiting the trans-cis isomerase of at least one visual cycle. In one embodiment, the cell is a Retinal Pigment Epithelial (RPE) cell.

In another embodiment, there is also provided a method of inhibiting a trans-cis isomerase enzyme of at least one visual cycle in a subject, the method comprising administering to the subject a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound having any of the structures of formulae (a) - (G) and (I) - (III) as described herein. In one embodiment, the individual is a human or non-human animal.

As used herein and in the appended claims, the singular forms "a", "and", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an agent" includes a plurality of such agents, and reference to "the cell" includes reference to one or more cells (or a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein to describe physical properties (e.g., molecular weight) or chemical properties (e.g., chemical formula), all combinations or subcombinations of ranges (subs) and specific embodiments thereof are intended to be included herein. When referring to a number or numerical range, the term "about" means that the number or numerical range referred to is an approximation within the experimental variability (or within statistical experimental error), and thus the number or numerical range may vary between 1% and 15% of the number or numerical range. The term "comprising" (and related terms such as "comprises" or "comprising" or "including") is not intended to exclude other certain embodiments, e.g., any combination of the substances, compositions, methods or steps described herein, as "consisting of" or "consisting essentially of" the recited features.

Incorporated by reference

All publications, patents and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference.

Drawings

The novel features believed characteristic of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

FIG. 1 depicts the concentration-dependent inhibition of isomerase activity by Compound 2 in the in vitro recombinant RPE65/LRAT system.

Figure 2 depicts the concentration-dependent inhibition of isomerase activity by compound 2 in vivo.

Figure 3 depicts the concentration-dependent inhibition of isomerase activity by compound 18 in vivo.

Figure 4 depicts the concentration-dependent inhibition of isomerase activity by compound 19 in vivo.

Figure 5 depicts the concentration-dependent inhibition of isomerase activity by compound 2 in vivo.

Figure 6 depicts the concentration-dependent inhibition of isomerase activity by compound 18 in vivo.

Figure 7 depicts the concentration-dependent inhibition of isomerase activity by compound 19 in vivo.

Figure 8 depicts the concentration-dependent inhibition of isomerase activity by compound 100 in vivo.

Figure 9 depicts the concentration-dependent inhibition of isomerase activity by compound 101 in vivo.

Figure 10 depicts the time course of the percent inhibition of isomerase activity by compound 2 in vivo.

Figure 11 depicts the time course of the percent inhibition of isomerase activity by compound 18 in vivo.

Figure 12 depicts the time course of the percent inhibition of isomerase activity by compound 19 in vivo.

Figure 13 depicts a single oral administration of compound 36 at a dose of 1 mg/kg.

Figure 14 depicts a single oral administration of compound 36 at a dose of 5 mg/kg.

Detailed Description

The alkynylphenyl derivative compounds described herein inhibit the isomerization step of the retinoid cycle. These compounds and compositions comprising these compounds may be useful for inhibiting degeneration of retinal cells or for increasing the survival of retinal cells. Thus, the compounds described herein may be beneficial in the treatment of ophthalmic diseases and disorders, such as age-related macular degeneration and stargardt disease.

Alkynyl phenyl derivative compound

In certain embodiments, alkynylphenyl derivative compounds are provided that include meta substitution end-capped with a nitrogen-containing moiety. The nitrogen-containing moiety may be, for example, an amine group (primary, secondary, and tertiary), an amide group, or an N-heterocyclic group. The linking atoms form a combination of linear structurally stable chemical bonds including carbon-carbon bonds, carbon-oxygen bonds, and the like.

In one embodiment is a compound having the structure of formula (a):

m is 0, 1, 2 or 3;

X is-O-, -S (= O)₂-、-N(R³¹)-、-C(=O)-、-C(=CH₂)-、-C(=N-NR³⁵) -OR-C (= N-OR)³⁵)-；

Y is a bond, -C (R)²⁷)(R²⁸) -or-C (R)²⁷)(R²⁸)-C(R²⁹)(R³⁰)-；

R²³and R²⁴Each independently selected from hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Performing the following steps; or R²³And R²⁴Together form an oxo group; or optionally, R ²³With adjacent R¹Together form a direct bond to provide a double bond; or optionally, R²³With adjacent R¹Together form a direct bond, and R²⁴With adjacent R²Together form a direct bond to provide a triple bond;

each R is¹⁴Are the same OR different and are each independently alkyl, halogen, fluoroalkyl OR-OR⁶；

R³⁰and R³⁵Each independently is hydrogen or C₁-C₅An alkyl group.

In yet another embodiment are compounds of formula (A) wherein R⁵Is an aryl group. In another embodiment are compounds of formula (A) wherein R⁵Is an unsaturated carbocyclic group. In yet another embodiment are compounds of formula (A) wherein R⁵Is a bicyclic carbocyclic group. In another embodiment compounds of formula (A) wherein R⁵Is norbornyl.

m is 0, 1, 2 or 3;

n is 0, 1, 2, 3, 4 or 5;

R¹and R²Each independently selected from hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Performing the following steps; or R¹And R²Together form an oxo group;

R³and R⁴Each independently selected from hydrogen or alkyl;

each R is⁷And each R⁸Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl or-C (= O) R⁹(ii) a Or R⁷And R⁸And together with the nitrogen atom to which they are attached form an N-heterocyclyl;

R¹⁵each being the same OR different and each being independently alkyl, -OR ⁶Alkenyl, alkynyl, halogen, fluoroalkyl, aryl, or aralkyl;

In yet another embodiment are compounds of formula (B) wherein R¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each independently hydrogen or alkyl.

In another embodiment are compounds of formula (B) wherein R¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each is hydrogen.

In another embodiment are compounds of formula (A) wherein R⁵Is 1-naphthyl or 2-naphthyl. In yet another embodiment are those wherein R¹、R²、R³、R⁴、R²³And R²⁴Each hydrogen. In another embodimentIn an embodiment, the compound is a compound of formula (a) wherein m is 0.

m is 0, 1, 2 or 3;

R¹and R²Each of which is the same or different and each is independently hydrogen, halogen, C₁-C₅Alkyl, fluoroalkyl, -OR⁶、-NR⁷R⁸Or a carbocyclic group; or R¹And R²Forming an oxo group;

each R is⁷And each R⁸Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl or-C (= O) R ⁹(ii) a Or R⁷And R⁸Together with the N atom to which they are attached form an N-heterocyclyl;

each R is⁹The same or different and are independently alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocycleA group;

R¹⁶、R¹⁷and R¹⁸Each being the same OR different and each being independently hydrogen, alkyl, -OR⁶Carbocyclyl or aryl.

In another embodiment are compounds of formula (C) wherein R¹²And R¹³Each is hydrogen. In another embodiment are compounds of formula (C) wherein R¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each independently hydrogen or alkyl. In another embodiment are compounds of formula (C) wherein m is 0, and R is ¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R³And R⁴Each is hydrogen. In another embodiment are compounds of formula (C) wherein R¹⁶、R¹⁷And R¹⁸Each independently is hydrogen, alkyl, carbocyclyl or aryl.

In another embodiment of the invention are compounds selected from the group consisting of: 4- (3- (3-aminopropyl) phenyl) but-3-yn-1-ol; 5- (3- (3-aminopropyl) phenyl) pent-4-yn-2-ol; 2- (3- (3-cyclopentylprop-1-ynyl) phenoxy) ethylamine; 3- (3- (3, 3-dimethylbut-1-ynyl) phenyl) propan-1-amine; 3- (3- (3-phenylprop-1-ynyl) phenyl) propan-1-amine; 3- (3- (pent-1-ynyl) phenyl) propan-1-amine; 3- (3- (hex-1-ynyl) phenyl) prop-1-amine; 3-amino-1- (3- (3-cyclopentylprop-1-ynyl) phenyl) propan-1-ol; 3-amino-1- (3- (3-phenylprop-1-ynyl) phenyl) propan-1-ol; 6- (3- (3-amino-1-hydroxypropyl) phenyl) hex-5-yn-1-ol; 4- (3- (3-amino-1-hydroxypropyl) phenyl) but-3-yn-1-ol; 3-amino-1- (3- (hept-1-ynyl) phenyl) propan-1-ol; 3- (3- (4-phenylbut-1-ynyl) phenyl) propan-1-amine; 3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol; 3- (3- (5-methoxypent-1-ynyl) phenyl) propan-1-amine; 3-amino-1- (3- (4-phenylbut-1-ynyl) phenyl) propan-1-ol; 6- (3- (3-aminopropyl) phenyl) hex-5-yn-1-ol; and 3- (3- (6-methoxyhex-1-ynyl) phenyl) prop-1-amine.

In another embodiment are compounds of formula (C) wherein R¹⁶is-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅Alkyl, and R¹⁷And R¹⁸Each independently hydrogen, alkyl or aryl.

In another embodiment are compounds selected from the group consisting of:

In another embodiment are compounds of formula (A) wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²) -, and R⁵Is a carbocyclyl group.

In another embodiment are compounds of formula (A) wherein R⁵Is cycloalkyl, Y is a bond, and the compound has the structure of formula (D):

m is 0, 1, 2 or 3;

p is 1, 2, 3, 4, 5 or 6;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

each R is³And R⁴Are the same or different and are each independently hydrogen or alkyl;

R¹²and R¹³Each of which is the same or different and is independently hydrogen, alkyl or-C (= O) R⁹(ii) a Or R¹²And R¹³Together with the N atom to which they are attached form an N-heterocyclyl;

In another embodiment are compounds of formula (D) wherein R¹²And R¹³Each is hydrogen. In another embodiment are compounds of formula (D) wherein R¹And R²Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently of the others is hydrogen, halogen, C₁-C₅Alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each independently hydrogen or alkyl. In another embodiment are compounds of formula (D) wherein p is 3, and R is ⁵Is a substituted or unsubstituted cyclopentyl group. In another embodiment are compounds of formula (D) wherein p is 4, and R is⁵Is a substituted or unsubstituted cyclohexyl group. In another embodiment are compounds of formula (D) wherein p is 5, and R is⁵Is a substituted or unsubstituted cycloheptyl. In another embodiment are compounds of formula (D) wherein R¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each is hydrogen. In another embodiment are compounds of formula (D) wherein m is 0. In another embodiment are compounds of formula (D) wherein q is 0. In another embodiment are compounds of formula (D) wherein q is 1, 2, 3, 4, or 5, and each R is¹⁹Independently is alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group.

In another embodiment is a compound selected from the group consisting of:

In another embodiment are compounds of formula (D) wherein R¹²Is hydrogen, and R¹³is-C (= O) R⁹Wherein R is⁹Is an alkyl group. In another embodiment are compounds of formula (D) wherein R¹And R²Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; r²³And R²⁴Each independently is hydrogen OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group; and R is³And R⁴Each is hydrogen. In another embodiment are compounds of formula (D) wherein m is 0. In another embodiment is a compound of claim 36, wherein q is 1, 2, 3, 4, or 5, and each R is¹⁹Independently is alkyl OR-OR⁶Wherein R is⁶Is hydrogen or C₁-C₅An alkyl group. In another embodiment are compounds of formula (D) wherein p is 4, and R is⁵Is a substituted or unsubstituted cyclohexyl group.

In another embodiment are compounds of formula (A) wherein Z is-C (R)²³)(R²⁴)-C(R¹)(R²) -, wherein R⁵Is a heterocyclic group and Y is a bond. In another embodiment are those wherein heterocyclyl may be substituted by-OR⁶Optionally substituted compound, wherein R⁶Is hydrogen or C₁-C₅An alkyl group. In another embodiment are those in which R ¹²And R¹³Each hydrogen.

m is 0, 1, 2 or 3;

R²¹and R²²Each independently is hydrogen, C₁-C₅Alkyl or fluoroalkyl;

R⁵is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R⁹is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

In another embodiment are compounds of formula (E) wherein R⁵Is an unsaturated carbocyclic group. In another embodiment are compounds of formula (E) wherein R⁵Is a bicyclic carbocyclic group. In another embodiment are compounds of formula (E) wherein R⁵A norbornyl group.

m is 0, 1, 2 or 3;

R²¹and R²²Each independently is hydrogen, C₁-C₅Alkyl or fluoroalkyl;

R⁹is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R¹⁶、R¹⁷、R¹⁸each being the same OR different and each being independently hydrogen, alkyl, -OR⁶Carbocyclyl or aryl.

In another embodiment are compounds of formula (F) wherein R¹²And R¹³Each is hydrogen. In another embodiment are compounds of formula (F) wherein R²¹、R²²、R³And R⁴Each independently is hydrogen or C₁-C₅An alkyl group. In another embodiment are compounds of formula (F) wherein m is 0. In another embodiment are compounds of formula (F) wherein R¹⁶、R¹⁷And R¹⁸Each independently hydrogen, alkyl OR-OR⁶Wherein each R is⁶Independently is hydrogen or C₁-C₅An alkyl group. In another embodiment are compounds of formula (F) wherein R ¹⁶、R¹⁷And R¹⁸Each independently hydrogen, alkyl or aryl.

In another embodiment is a compound selected from the group consisting of: 4- ((3- (2-aminoethoxy) phenyl) ethynyl) heptan-4-ol; 1- (3- (2-aminoethoxy) phenyl) -3-ethylpent-1-yn-3-ol; 1- (3- (2-aminoethoxy) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol; 5- ((3- (2-aminoethoxy) phenyl) ethynyl) nonan-5-ol; 4- (3- (2-aminoethoxy) phenyl) -2-methylbut-3-yn-2-ol; 2- (3- (hept-1-ynyl) phenoxy) ethylamine; 4- (3- (2-aminoethoxy) phenyl) but-3-yn-1-ol; 2- (3- (3-phenylprop-1-ynyl) phenoxy) ethylamine; 2- (3- (4-methylpent-1-ynyl) phenoxy) ethylamine; 6- (3- (2-aminoethoxy) phenyl) hex-5-yn-1-ol; 2- (3- (3-ethylpent-1-ynyl) phenoxy) prop-1-amine; 2- (3- (3-propylhex-1-ynyl) phenoxy) prop-1-amine; 1- (3- (2-aminoethoxy) phenyl) -3-ethylpent-1-yn-3-ol; 4- ((3- (1-aminopropyl-2-yloxy) phenyl) ethynyl) hept-4-ol; 2- (3- (3-ethylpent-1-ynyl) phenoxy) ethylamine; and 2- (3- (3-propylhex-1-ynyl) phenoxy) ethylamine.

m is 0, 1, 2 or 3;

p is 1, 2, 3, 4 or 5;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

R⁹is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

In another embodiment is a non-retinoid compound that inhibits the isomerase reaction that results in the production of 11-cis-retinol, wherein the isomerase reaction occurs in the RPE and the compound has an ED50 value of less than 1mg/kg when administered to an individual. In another embodiment is a non-retinoid compound, wherein the ED50 value is measured after a single dose of the compound is administered to the individual for about 2 hours or more. In yet another embodiment, the compound is an amine compound having an alkynylphenyl group attached. In yet another embodiment, the compound is a non-retinoid compound.

In another embodiment, a method of inhibiting a trans-cis isomerase of at least one visual cycle in a cell is provided, wherein the method comprises contacting the cell with a compound having any of structures (a) - (G) as described herein, thereby inhibiting the trans-cis isomerase of at least one visual cycle. In one embodiment, the cell is a Retinal Pigment Epithelial (RPE) cell.

In particular embodiments of the methods described above and herein, the accumulation of lipofuscin pigment in the eye of the subject is inhibited, and in certain embodiments, the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E). In certain other embodiments, degeneration of retinal cells is inhibited. In a specific embodiment, the retinal cell is a retinal nerve cell, wherein the retinal nerve cell is a photoreceptor cell, an amacrine cell, a horizontal cell, a ganglion cell, or a bipolar cell. In another specific embodiment, the retinal cell is a Retinal Pigment Epithelium (RPE) cell.

In addition, compounds having the structure of formula (I):

m is 0, 1, 2 or 3;

R₁And R₂Forming an oxo group;

R₃and R₄Each is the same or different, andeach independently is hydrogen or alkyl;

R₅is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₆is hydrogen or alkyl;

R₇and R₈Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl or-C (= O) R₉(ii) a Or

R₇And R₈And together with the nitrogen atom to which they are attached form an N-heterocyclyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

x is-C (R)₁₀)(R₁₁) -or-O-;

R₁₀And R₁₁Forming an oxo group;

each R is₁₄Are the same OR different and are each independently alkyl, halogen, fluoroalkyl OR-OR₆。

In certain embodiments, X is-C (R)₁₀)(R₁₁) -, and the compounds of the general formula (I) have a propylene linkage. Thus, the compound may be represented by the structure of general formula (II):

one embodiment of the present invention provides a compound having the structure of formula (II) wherein R₅Is aryl, as defined herein.

In another embodiment, the present invention provides a compound having the structure of formula (II), wherein R₅Is phenyl. The compound may be represented by the structure of formula (IIa):

m is 0, 1, 2 or 3;

n is 0, 1, 2, 3, 4 or 5;

R₁and R₂Each being the same OR different and each being independently hydrogen, halogen, alkyl, fluoroalkyl, -OR₆、-NR₇R₈Or in a carbocyclic group; or

R₁And R₂Forming an oxo group;

R₆Is hydrogen or alkyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₀and R₁₁Each being the same OR different and each being independently hydrogen, halogen, alkyl, fluoroalkyl, -OR₆、-NR₇R₈Or a carbocyclic group; or R₁₀And R₁₁Forming an oxo group;

R₁₂and R₁₃Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R₉(ii) a Or R₁₂And R₁₃And together with the nitrogen atom to which they are attached form an N-heterocyclyl;

each R is₁₄Are the same OR different and are each independently alkyl, halogen, fluoroalkyl OR-OR₆(ii) a And

each R is₁₅Are the same OR different and are each independently alkyl, -OR₆Alkenyl, alkynyl, halogen, fluoroalkyl, aryl or aralkyl.

In certain embodiments, R₁₂And R₁₃Each is hydrogen. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently hydrogen, halogen, alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently is hydrogen OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, m is 0, n is 0, 1 or 2, and each R is ₁₅Independently is alkyl, -OR₆Or an aryl group.

In certain embodiments, the compounds of formula (I), (II), or (IIa) have the structures shown in table 1.

TABLE 1

In another embodiment, compounds of formula (II) are provided wherein R₅Is naphthyl. In yet another embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each is hydrogen.

In particular embodiments, the compounds of formula (I) or (II) have the structures shown in table 2.

TABLE 2

In a further embodiment of the invention, there are provided compounds of the general formula (II) wherein R₅Is an alkyl group. Thus, the compound may be represented by the structure of formula (IIb):

m is 0, 1, 2 or 3;

R₁And R₂Forming an oxo group;

R₆is hydrogen or alkyl;

R₇and R₈Each of which is the same or different and each is independently hydrogen, alkyl, carbocyclyl or-C (= O) R ₉(ii) a Or

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl; r₁₀And R₁₁Each being the same OR different and each being independently hydrogen, halogen, alkyl, fluoroalkyl, -OR₆、-NR₇R₈Or a carbocyclic group; or

R₁₀And R₁₁Forming an oxo group;

R₁₂and R₁₃Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R₉(ii) a Or

R₁₂And R₁₃Together with the nitrogen atom to which they are attached form an N-heterocyclyl;

R₁₆、R₁₇and R₁₈Each being the same OR different and each being independently hydrogen, alkyl, -OR₆Carbocyclyl or aryl.

In certain embodiments, R₁₂And R₁₃Each is hydrogen. In certain embodiments, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently hydrogen, halogen, alkyl OR-OR₆Wherein R is₆Is hydrogen or an alkaneAnd (4) a base. In certain embodiments, m is 0, and R is₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently is hydrogen OR-OR₆Wherein R is₆Is hydrogen or alkyl. In yet another embodiment, R₁₆、R₁₇And R₁₈Each independently is hydrogen, alkyl, carbocyclyl or aryl.

In certain embodiments, the compounds of formula (I), (II), or (IIb) have the structures shown in table 3.

TABLE 3

In another embodiment, R₁₆is-OR₆Wherein R is₆Is hydrogen or alkyl, and R₁₇And R₁₈Each independently hydrogen, alkyl or aryl.

In yet another specific embodiment, the compound of formula (I), (II) or (IIb) has the structure shown in Table 4.

TABLE 4

In certain embodiments, R₁₂Is hydrogen, and R₁₃is-C (= O) R₉Wherein R is₉Is an alkyl group. One particular embodiment provides a compound: n- (3-3 (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide. In yet another embodiment, compounds having the structure of formula (II) are provided wherein R₅Is a carbocyclyl group.

In certain embodiments, R₅Is cycloalkyl, and the compound may be represented by the structure of formula (IIc):

m is 0, 1, 2 or 3;

p is 1, 2, 3, 4 or 5;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

R₁And R₂Forming an oxo group;

R₃and R ₄Each is the same or different and each is independently hydrogen or alkyl;

R₆is hydrogen or alkyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₀And R₁₁Forming an oxo group;

each R is₁₄The same OR different and independently is alkyl, halogen, fluoroalkyl OR-OR₆(ii) a And

each R is₁₉Are the same OR different and are independently alkyl, -OR₆Halogen or fluoroalkyl.

In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently hydrogen, halogen, alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently is hydrogen OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, m is 0. In one embodiment, q is 0. In yet another embodiment, q is 1, 2, 3, 4 or 5, and each R ₁₉Independently is alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In yet another embodiment, p is 1 and R₅Is cyclopropyl. In yet another embodiment, p is 2, and R is₅Is a cyclobutyl group. In yet another embodiment, p is 3, and R₅Is cyclopentyl. In another embodiment, p is 4, and R is₅Is cyclohexyl. In other embodiments, p is 5, and R is₅Is cycloheptyl.

In a particular embodiment, the compounds of formula (I), (II) or (IIc) have the structures shown in table 5.

TABLE 5

In certain embodiments, R₁₂Is hydrogen, and R₁₃is-C (= O) R₉Wherein R is₉Is an alkyl group. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently is hydrogen OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, m is 0. In yet another embodiment, q is 0. In yet another embodiment, q is 1, 2, 3, 4 or 5, and each R is₁₉Independently is alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In another embodiment, p is 1 and R₅Is cyclopropyl. In yet another embodiment, p is 2, and R is₅Is a cyclobutyl group. In yet another embodiment, p is 3, and R₅Is cyclopentyl. In another embodiment, p is 4, and R is ₅Is cyclohexyl. In other embodiments, p is 5, and R is₅Is cycloheptyl.

In particular embodiments, the compounds of formula (I), (II), or (IIc) have the structures shown in table 6.

TABLE 6

In yet another embodiment, compounds having the structure of formula (II) are provided wherein R₅Is a heterocyclic group, as defined herein. In one embodiment, the heterocyclic group may optionally be-OR₆Is substituted in which R₆Is hydrogen or alkyl. In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, m is 0. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently is hydrogen, halogen, alkyl or-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each is hydrogen.

In one embodiment, the compounds of formula (I) or (II) have the structures shown in table 7.

TABLE 7

In yet another embodiment, compounds having the structure of formula (II) are provided wherein R₅Is heteroaryl, as defined herein. In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, m is 0. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently hydrogen, halogen, alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, R ₁、R₂、R₃、R₄、R₁₀And R₁₁Each is hydrogen.

In a particular embodiment, the compounds of formula (I) or (II) have the structures shown in table 8.

TABLE 8

In yet another embodiment, there is provided a compound of formula (I) wherein X is-O-and the compound has an ethylene oxide linkage. Thus, the compound may be represented by the structure of formula (III):

m is 0, 1, 2 or 3;

R₁and R₂Each of which is the same or different and each is independently hydrogen, alkyl or fluoroalkyl; or R₁And R₂Forming an oxo group;

R₅is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₆is hydrogen or alkyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₂and R₁₃Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R₉(ii) a Or R₁₂And R₁₃Together with the nitrogen atom to which they are attached form an N-heterocyclyl; and

each R is₁₄The same OR different and independently is alkyl, halogen, fluoroalkyl OR-OR ₆。

In one embodiment, R₅Is an alkyl group, and the compound has the structure of formula (IIIa):

m is 0, 1, 2 or 3;

R₆is hydrogen or alkyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₂and R₁₃Each of which is the same or different and each is independently hydrogen, alkyl or-C (= O) R₉(ii) a Or R₁₂And R₁₃Together with the nitrogen atom to which they are attached form an N-heterocyclyl;

In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, m is 0. In one embodiment, R₁、R₂、R₃And R₄Each independently hydrogen or alkyl. In another embodiment, R ₁₆、R₁₇And R₁₈Each independently hydrogen, alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In yet another embodiment, R₁₆is-OR₆Wherein R is₆Is hydrogen or alkyl, and R₁₇And R₁₈Each independently is an alkyl group. In other embodiments, R₁₆、R₁₇And R₁₈Each independently hydrogen or aryl. In other embodiments, R₁₆、R₁₇And R₁₈Each independently hydrogen or alkyl.

In another specific embodiment, the compound of formula (I), (III) or (IIIa) has the structure shown in table 9.

TABLE 9

In yet another embodiment, compounds of the structure of formula (III) are provided wherein R₅Is a carbocyclyl group.

In one embodiment, R₅Is cycloalkyl, and the compound may be represented by the structure of formula (IIIb):

m is 0, 1, 2 or 3;

p is 1, 2, 3, 4 or 5;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

R₆Is hydrogen or alkyl;

R₉is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, m is 0. In one embodiment, R₁、R₂、R₃、R₄、R₁₀And R₁₁Each independently hydrogen, halogen, alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In one embodiment, R₁、R₂、R₃And R₄Each independently hydrogen or alkyl. In another embodiment, q is 1, 2, 3, 4 or 5, and each R is₁₉Independently is alkyl OR-OR₆Wherein R is₆Is hydrogen or alkyl. In another embodiment, q is 0. In yet another embodiment, q is 1 and R₁₉is-OR₆Wherein R is₆Is hydrogen or alkyl. In another embodiment, p is 1 and R₅Is cyclopropyl. In other embodiments, p is 2, and R is₅Is a cyclobutyl group. In other embodiments, p is 3, and R ₅Is cyclopentyl. In other embodiments, p is 4, and R₅Is cyclohexyl. In other embodiments, p is 5, and R is₅Is cycloheptyl.

In a particular embodiment, the compounds of formula (I), (III) or (IIIb) have the structures shown in table 10.

Watch 10

Another embodiment provides a compound having the structure of formula (III), wherein R₅Is heteroaryl. In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, m is 0. In one embodiment, R₁、R₂、R₃And R₄Each independently hydrogen or alkyl.

In a particular embodiment, the compounds of formula (I) or (III) have the structures shown in table 11.

TABLE 11

In yet another embodiment, the present invention provides a compound having the structure of formula (III), wherein R₅Is an aryl group. In one embodiment, R₁₂And R₁₃Each is hydrogen. In one embodiment, m is 0. In one embodiment, R₁、R₂、R₃And R₄Each independently hydrogen or alkyl.

In a particular embodiment, the compounds of formula (I) or (III) have the structures shown in table 12.

TABLE 12

In another specific embodiment, the compounds of formulae (a) - (G) and (I) - (III) have the structures shown in table 13.

Watch 13

Definition of

As used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes a plurality of such compounds, and reference to "the cell" includes reference to one or more cells (or a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein to describe physical properties (e.g., molecular weight) or chemical properties (e.g., chemical formula), all combinations or subcombinations of ranges (subs) and specific embodiments thereof are intended to be included herein. The term "about" when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within the experimental range of variation (or within statistical experimental error), and thus the number or numerical range may vary between 1% and 15% of the number or numerical range. The term "comprising" (and related terms such as "comprises" or "comprising," or "includes") is not intended to exclude other certain embodiments, e.g., embodiments of any combination of the substances, compositions, methods or steps described herein, and the like, from "consisting of" or "consisting essentially of" the recited features.

"amino" refers to-NH₂A group.

"cyano" refers to the group-CN.

"Nitro" means-NO₂A group.

"oxy" refers to the-O-group.

"oxo" refers to an = O group.

"imino" refers to an = NH group.

"hydrazino" means = N-NH₂A group.

"thio" refers to an = S group.

"alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from 1 to 15 carbon atoms (e.g., C)₁-C₁₅Alkyl groups). In one embodiment, the alkyl group contains 1 to 13 carbon atoms (e.g., C)₁-C₁₃Alkyl groups). In certain embodiments, the alkyl group contains 1-8 carbon atoms (e.g., C)₁-C₈Alkyl groups). In other embodiments, the alkyl group includes 5 to 15 carbon atoms (e.g., C)₅-C₁₅Alkyl groups). In certain embodiments, the alkyl group contains 5 to 8 carbon atoms (e.g., C)₅-C₈Alkyl groups). The alkyl group is bonded to the rest of the molecule by a single bond, for example, methyl (Me), ethyl (Et), n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, and the like. Unless otherwise specifically stated in the specification, an alkyl group is optionally substituted with one or more of the following substituents: halogen, cyano, nitro, oxo, thio, trimethylsilyl, -OR ^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -S (O)_tOR^a(wherein t is 1 or 2) and-S (O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently is hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl, or heteroaralkyl.

"alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having from 2 to 12 carbon atoms. In certain embodiments, alkenyl groups contain 2 to 8 carbon atoms. In other embodiments, the alkenyl group contains 2 to 4 carbon atoms. The alkenyl group is attached to the remainder of the molecule by a single bond, for example, vinyl (i.e., vinyl), 1-propenyl (i.e., allyl), 1-butenyl, 1-pentenyl, 1, 4-pentadienyl, and the like. Unless otherwise specifically stated in the specification, an alkenyl group is optionally substituted with one or more of the following substituents: halogen, cyano, nitro, oxo, thio, trimethylsilyl, -OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -S (O)_tOR^a(wherein t is 1 or 2) and-S (O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently is hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl, or heteroaralkyl.

"alkynyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having from 2 to 12 carbon atoms. In certain embodiments, alkynyl groups contain 2-8 carbon atoms. In other embodiments, alkynyl groups have 2-4 carbon atoms. The alkynyl group is bonded to the rest of the molecule by a single bond, and examples thereof include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless otherwise specifically stated in the specification, the alkynyl group is optionally substituted by one or more substituents selected from: halogen, cyano, nitro, oxo, thio, trimethylsilyl, -OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -S (O)_tOR^a(wherein t is 1 or 2) and-S (O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently is hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl, or heteroaralkyl.

"alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain connecting the remainder of the molecule to a group, consisting solely of carbon and hydrogen, containing no unsaturation, and having from 1 to 12 carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, and the like. The The alkylene chain is attached to the rest of the molecule by single bonds and to the group by single bonds. The point at which the alkylene chain is attached to the remainder of the molecule and the point at which the alkylene chain is attached to the group may be one carbon atom in the alkylene chain or through any two carbon atoms in the chain. Unless otherwise specifically stated in the specification, the alkylene chain is optionally substituted with one or more of the following substituents: halogen, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, thio, trimethylsilyl, -OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -S (O)_tOR^a(wherein t is 1 or 2) and-S (O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently is hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl, or heteroaralkyl.

"alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain, consisting solely of carbon and hydrogen, containing at least one double bond, and having from 2 to 12 carbon atoms, e.g., ethenylene, propenylene, n-propenyl, and the like, linking the remainder of the molecule to a group. The alkenylene chain is attached to the rest of the molecule by a double or single bond and to the group by a double or single bond. The point at which the alkenylene chain is attached to the rest of the molecule and the point at which the alkenylene chain is attached to the group may be one carbon atom or any two carbon atoms in the chain. Unless otherwise specifically stated in the specification, an alkenylene chain is optionally substituted with one or more of the following substituents: halogen, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, thio, trimethylsilyl, -OR ^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -S (O)_tOR^a(wherein t is 1 or 2) and-S (O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently is hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl (cycloalkylalkyl), aryl (optionally substituted with one or more halo groups), aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl, or heteroaralkyl, and each of the foregoing substituents is unsubstituted, unless otherwise specifically indicated.

"aryl" refers to a group derived from an aromatic monocyclic or polycyclic hydrocarbon ring system by the removal of a hydrogen atom from a ring carbon atom. Aromatic mono-or polycyclic hydrocarbon ring systems contain only hydrogen and 6 to 18 carbon atoms, wherein at least one ring is fully unsaturated in the ring system, i.e. it contains rings which are delocalized according to houckel theory to (4n +2) pi-electron systems. Aryl groups include, but are not limited to, the following: for example, phenyl, fluorenyl, and naphthyl. Unless otherwise specifically stated in the specification, the term "aryl" or the prefix "ar-" (e.g., "aralkyl") is meant to include aryl groups optionally substituted with one or more substituents independently selected from: alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocycloalkyl, optionally substituted heterocyclyl, optionally substituted heterocycloalkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -R ^b-OR^a、-R^b-OC(O)-R^a、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -R^b-S(O)_tOR^a(wherein t is 1 or 2) and-R^b-S(O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl (optionally substituted with one or more halo groups), aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl or heteroaralkyl, R^bEach independently being a direct bond, or a linear or branched alkylene or alkenylene chain, and R^cIs a straight or branched alkylene or alkenylene chain, and wherein, unless otherwise stated, each of the above substituents is unsubstituted.

"aralkyl" refers to the general formula-R^c-a radical of an aryl radical, wherein R^cAs an alkylene chain as defined above, for example, benzyl, diphenylmethyl, and the like. As described above for the alkylene chain, the alkylene chain portion of the aralkyl group is optionally substituted. As described above for aryl, the aryl portion of an aralkyl group is optionally substituted.

"aralkenyl" refers to the general formula-R^dA radical of aryl, wherein R^dIs an alkenylene chain as described above. As described above for aryl, the aryl portion of the aralkenyl is optionally substituted. As described above for alkenylene, the alkenylene chain portion of the aralkenyl group is optionally substituted.

"aralkynyl" refers to the formula-R^eA radical of aryl, wherein R^eIs an alkynylene chain as defined above. As described above for aryl, the aryl portion of an arylalkynyl group is optionally substituted. As described above for the alkynylene chain, the alkynylene chain portion of the arylalkynyl group is optionally substituted.

"carbocyclyl" refers to a stable non-ring consisting solely of carbon and hydrogen atomsAromatic monocyclic or polycyclic hydrocarbon groups, including fused or bridged ring systems, containing from 3 to 15 carbon atoms. In certain embodiments, carbocycles include 3-10 carbon atoms. In other embodiments, carbocyclyl contains 5 to 7 carbon atoms. The carbocyclyl group is attached to the remainder of the molecule by a single bond. Carbocyclyl groups are optionally saturated (i.e., contain only C-C single bonds) or unsaturated (i.e., contain one or more double or triple bonds). A fully saturated carbocyclic group is also referred to as a "cycloalkyl". Examples of monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Unsaturated carbocyclyl groups are also known as "cycloalkenyl". Examples of monocyclic cycloalkenyl groups include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl groups include, for example, adamantyl, norbornyl (i.e., bicyclo [ 2.2.1) ]Heptylalkyl), norbornenyl, naphthylalkyl, 7-dimethyl-bicyclo [2.2.1]Heptalkyl, and the like. Unless otherwise specifically stated in the specification, the term "carbocyclyl" is meant to include carbocyclyl optionally substituted with one or more substituents independently selected from: alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, oxo, thio, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocycloalkyl, optionally substituted heterocyclyl, optionally substituted heterocycloalkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -R^b-OR^a、-R^b-SR^a、-R^b-OC(O)-R^a、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -R^b-S(O)_tOR^a(wherein t is 1 or 2) and-R^b-S(O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl or heteroaralkyl, R^bEach independently being a direct bond or a linear or branched alkylene or alkenylene chain, and R ^cAre straight or branched alkylene or alkenylene chains, and the substituents described above are each unsubstituted, unless otherwise specified.

"Carbocycloalkyl" refers to the general formula-R^cA carbocyclic group, wherein R^cIs an alkylene chain as defined above. The alkylene chain and carbocyclyl are optionally substituted as defined above.

"halo" refers to a bromo, chloro, fluoro, or iodo substituent.

"fluoroalkyl" refers to an alkyl group, as defined above, i.e., substituted with one or more fluoro groups, as defined above, e.g., trifluoromethyl, difluoromethyl, 2,2, 2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. As defined above for alkyl, the alkyl portion of fluoroalkyl is optionally substituted.

"Heterocyclyl" refers to a stable 3 to 18-membered non-aromatic cyclic group containing 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur. Unless otherwise specifically stated in the specification, the heterocyclic group is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, and includes a fused or bridged ring system. The heteroatoms in the heterocyclic group are optionally oxidized. If one or more nitrogen atoms are present, are optionally quaternized. The heterocyclic group is partially or fully saturated. The heterocyclic group is attached to the remainder of the molecule through any of the ring atoms. Examples of such heterocyclic groups include, but are not limited to, dioxolanyl, thienyl [1,3 ] ]Dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidinonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, and 1, 1-dioxo-thiomorpholinyl. Unless otherwise specifically stated in the specification, the term "heterocyclyl" is meant to include heterocyclyl groups as defined above optionally substituted with one or more substituents selected from: alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, oxo, thio, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocycloalkyl, optionally substituted heterocyclyl, optionally substituted heterocycloalkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -R ^b-OR^a、-R^b-SR^a、-R^b-OC(O)-R^a、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -R^b-S(O)_tOR^a(wherein t is 1 or 2) and-R^b-S(O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl or heteroaralkyl, R^bEach independently being a direct bond or a linear or branched alkylene or alkenylene chain, and R^cAre straight or branched alkylene or alkenylene chains, and the substituents described above are each unsubstituted, unless otherwise specified.

"N-heterocyclyl" or "N-linked heterocyclyl" refers to a heterocyclyl group as defined above that contains at least one nitrogen, wherein the point at which the heterocyclyl group is attached to the remainder of the molecule is a nitrogen atom in the heterocyclyl group. As described above for heterocyclyl, the N-heterocyclyl group is optionally substituted. Examples of such N-heterocyclyl groups include, but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

"C-heterocyclyl" or "C-linked heterocyclyl" refers to a heterocyclyl group, as defined above, that contains at least one heteroatom and the point of attachment of the heterocyclyl group to the rest of the molecule is through a carbon atom in the heterocyclyl group. As described above for heterocyclyl, C-heterocyclyl is optionally substituted. Examples of such C-heterocyclyl groups include, but are not limited to, 2-morpholinyl, 2-or 3-or 4-piperidinyl, 2-piperazinyl, 2-or 3-pyrazolidinyl, and the like.

"Heterocycloalkyl" refers to the general formula-R^cA group of heterocyclic groups, wherein R^cIs an alkylene chain as defined above. If the heterocyclic group is a nitrogen-containing heterocyclic group, the heterocyclic group is optionally attached to an alkyl group at the nitrogen atom. As defined above for the alkylene chain, the alkylene chain of the heterocycloalkyl group is optionally substituted. As defined above for heterocyclyl, the heterocyclyl portion of a heterocycloalkyl group is optionally substituted.

"heteroaryl" refers to a group derived from a 3-to 18-membered aromatic ring group containing 2-17 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, heteroaryl is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the ring systems is fully unsaturated, i.e., it comprises a ring that is delocalized to a (4n +2) pi-electron system according to the theory of shockl. Heteroaryl groups include fused or bridged ring systems. The heteroatoms in the heteroaryl group are optionally oxidized. If one or more nitrogen atoms are present, optionally quaternized. The heteroaryl group is attached to the rest of the molecule through any atom in the ring. Heteroaryl radicalExamples of (b) include, but are not limited to, azanyl (azepinoyl), acridinyl, benzimidazolyl, benzindolyl, 1, 3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo [ d ] ]Thiazolyl, benzothiadiazolyl, benzo [ b ]][1,4]Dioxaheptatrienes, benzo [ b][1,4]Oxazinyl, 1, 4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothienyl), benzothieno [3,2-d]Pyrimidinyl, benzotriazolyl, benzo [4,6 ]]Imidazo [1,2-a ]]Pyridyl, carbazolyl, cinnolinyl, cyclopenta [ d ]]Pyrimidinyl, 6, 7-dihydro-5H-cyclopenta [4,5 ]]Thieno [2,3-d ]]Pyrimidinyl, 5, 6-dihydrobenzo [ h ]]Quinazolinyl, 5, 6-dihydrobenzo [ h ]]Cinnolinyl, 6, 7-dihydro-5H-benzo [6,7 ]]Cyclohepta [1,2-c ]]Pyridazine, dibenzofuranyl, dibenzothienyl, furanyl, furanonyl, fluoro [3,2-c ]]Pyridyl, 5,6,7,8,9, 10-hexahydrocycloocta [ d ]]Pyrimidinyl, 5,6,7,8,9, 10-hexahydrocyclooctane [ d ]]Pyridazinyl, 5,6,7,8,9, 10-hexahydrocyclooctane [ d ]]Pyridyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolinyl, indolizinyl, isoxazolyl, 5, 8-methylene-5, 6,7, 8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl (1,6-naphthyridinonyl), oxadiazolyl, 2-oxaza, oxazolyl, oxetanyl, 5,6,6a,7,8,9,10,10 a-octahydrobenzo [ h ] h ]Quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo [3,4-d]Pyrimidinyl, pyridinyl, pyrido [3,2-d ]]Pyrimidinyl, pyrido [3,4-d ]]Pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7, 8-tetrahydroquinazolinyl, 5,6,7, 8-tetrahydrobenzo [4,5 ] tetrahydroquinoline]Thieno [2,3-d ]]Pyrimidinyl, 6,7,8, 9-tetrahydro-5H-cyclohepta [4,5 ]]Thieno [2,3-d ]]Pyrimidinyl, 5,6,7, 8-tetrahydropyrido [4,5-c]Pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno [2,3-d ]]Pyrimidinyl, thieno [3 ],2-d]Pyrimidinyl, thieno [2, 3-c)]Pyridyl and thienyl (i.e., thienyl). Unless otherwise specifically stated in the specification, the term "heteroaryl" is meant to include heteroaryl groups as defined above optionally substituted with one or more substituents selected from: alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thio, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocycloalkyl, optionally substituted heterocyclyl, optionally substituted heterocycloalkyl, optionally substituted heteroaryl, optionally substituted heteroaralkyl, -R ^b-OR^a、-R^b-SR^a、-R^b-OC(O)-R^a、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a(wherein t is 1 or 2), -R^b-S(O)_tOR^a(wherein t is 1 or 2) and-R^b-S(O)_tN(R^a)₂(wherein t is 1 or 2) wherein R^aEach independently is hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocycloalkyl, heteroaryl, or heteroaralkyl, wherein R is^bEach independently being a direct bond or a linear or branched alkylene or alkenylene chain, and R^cAre straight or branched alkylene or alkenylene chains, and the substituents described above are each unsubstituted, unless otherwise specified.

"N-heteroaryl" refers to a heteroaryl group as defined above that contains at least one nitrogen atom, and wherein the point at which the heteroaryl group is attached to the remainder of the molecule is the nitrogen atom in the heteroaryl group. As defined above for heteroaryl, the N-heteroaryl group is optionally substituted.

"C-heteroaryl" refers to a heteroaryl group as defined above, and the point of attachment of the heteroaryl group to the rest of the molecule is through a carbon atom in the heteroaryl group. As described above for heteroaryl, C-heteroaryl is optionally substituted.

"Heteroaralkyl" refers to the general formula-R^c-a heteroaryl group, wherein R^cIs an alkylene chain as defined above. If the heteroaryl group is a nitrogen-containing heteroaryl group, the heteroaryl group is attached to the alkyl group at the nitrogen atom. As defined above for the alkylene chain, the alkylene chain of the heteroaralkyl group is optionally substituted. As defined above for the heteroaryl group, the heteroaryl portion of the heteroaralkyl group is optionally substituted.

The compounds or their pharmaceutically acceptable salts may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers and other stereoisomeric forms which, depending on the absolute stereochemistry, may be defined as (R) -or (S) -or (D) -or (L) -for amino acids. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both E and Z geometric isomers (e.g., cis or trans). Likewise, all possible isomers and their racemic and optically pure forms, as well as all tautomeric forms are also intended to be included.

"stereoisomers" refers to compounds consisting of identical atoms bonded by the same bonds, but having different three-dimensional structures, which are not interconvertible. Thus, various stereoisomers and mixtures thereof are considered to include "enantiomers," which refers to two stereoisomers whose molecules are non-overlapping mirror images of each other.

"tautomer" refers to the transfer of a proton from one atom of a molecule to another atom of the same molecule. The compounds presented herein may exist as tautomers. Tautomers are compounds that can interconvert by the migration of a hydrogen atom, which is accompanied by the conversion of a single bond and an adjacent double bond. In solutions where tautomerization may be present, the chemical equilibrium of the tautomers will exist. The exact ratio of tautomers depends on a number of factors including temperature, solvent and pH. Some examples of tautomeric pairs include:

"optional" or "optionally" means that the subsequently described activity or circumstance may or may not occur, and that the description includes instances where the activity or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" refers to aryl substituted or unsubstituted, and the description includes both substituted aryl and unsubstituted aryl.

"pharmaceutically acceptable salts" include both acid and base addition salts. The pharmaceutically acceptable salts of any of the amine derivative compounds described herein to which an alkynylphenyl group is attached include any and all pharmaceutically suitable salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"pharmaceutically acceptable acid addition salts" refers to those salts that retain biological effectiveness and free basic properties, are not biological products, are otherwise undesirable, and are formed with inorganic acids (e.g., hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, hydroiodic, hydrofluoric, phosphorous, and the like). Also included are salts formed with organic acids such as aliphatic mono-and dicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like, as well as including, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Exemplary salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfonate, and the like. Also contemplated are salts of amino acids such as arginine, gluconate, and galacturonate (see, e.g., Berges. metals, "pharmaceutical salts," journal of pharmaceutical science,66:1-19(1997), which are incorporated herein by reference in their entirety). Acid addition salts of base compounds can be prepared by contacting the free base form with a sufficient amount of the desired acid according to methods and techniques well known to those skilled in the art.

"pharmacologically acceptable base addition salts" refers to those salts that retain biological effectiveness and free acidic properties, which are not biologicals, otherwise undesirable. These salts are prepared by adding an inorganic or organic base to the free acid. Pharmaceutically acceptable base addition salts may be formed with metals or amines, for example alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines; substituted amines (including naturally occurring substituted amines); salts of cyclic amines with basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methyl reduced glucamine, glucosamine, methyl reduced glucamine, theobromine (theobromamine), purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. See bergetial, supra.

"non-retinoid compound" refers to any compound that is not a retinoid. Retinoids refer to compounds having a diterpene backbone containing a trimethylcyclohexenyl ring and a polyene chain terminated with polar end groups. Examples of retinoids include retinal and derived imines/hydrazides/oximes, retinol and optionally derived esters, retinylamine and optionally derived amides, retinoic acid and optionally derived esters or amides. The non-retinoid compound may include an optional internal cyclic group (e.g., aromatic group). The non-retinoid compound may include an optional amine group with an alkynylphenyl group attached.

As used herein, "treat" or "ameliorate" or "improve" may be used interchangeably. These terms refer to methods of achieving beneficial or desired results, including but not limited to therapeutic and/or prophylactic effects. Therapeutic effect refers to the elimination or amelioration of the underlying disorder being treated. Likewise, the therapeutic effect is achieved by the elimination or amelioration of one or more physiological characteristics associated with the underlying disorder, such that an improvement in the condition is observed in the individual, even though the individual is still afflicted with the underlying disorder. With respect to prophylactic efficacy, the compositions can be administered to an individual at risk of developing a particular disease, or to an individual exhibiting one or more physiological characteristics of a disease, even if the disease has not yet been diagnosed.

"prodrug" refers to a compound that is converted under physiological conditions or by solvolysis to a biologically active compound as described herein. Thus, the term "prodrug" refers to a precursor of a pharmaceutically acceptable biologically active compound. When administered to an individual, the prodrug may be inactive, but converted in vivo to an active compound, e.g., by hydrolysis. Such prodrug compounds generally have the advantage of solubility, histocompatibility or delayed release in mammals (see, e.g., Bundgard, h., designnof produgs (1985), pp.7-9,21-24(Elsevier, Amsterdam)).

A discussion of prodrugs is provided in Higuchi, T.et al, "Pro-drugs NovelDelivery systems," A.C.S.Symphosium series, Vol.14 and BioversibleCrierinsinDrugDesign, ed.Edward dB. Roche, American pharmaceutical Association and PergammonPress, 1987, which are incorporated herein by reference in their entirety.

The term "prodrug" is also meant to include covalently bonded carriers that release the active compound in vivo when such prodrug is administered to a mammalian subject. Prodrugs of the active compounds described herein may be prepared by modifying functional groups present in the active compound by: the variants are isolated as the parent active compound, either in routine manipulation or in vivo. Prodrugs include compounds wherein a hydroxy, amino, or sulfhydryl group is attached to any group that, when the prodrug of the active compound is administered to a mammalian subject, separates to form a free hydroxy, free amino, or free sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohols or acetate, formate and benzoate derivatives of amine functional groups in the active compound, and the like.

Preparation of alkynylphenyl derivative compounds

In general, the compounds used in the reactions described herein can be prepared according to organic synthesis techniques well known to those skilled in the art, starting from commercially available chemicals and/or from compounds described in the chemical literature. "commercially available chemicals" are available from reliable commercial sources and include Across organics (Pittsburgh PA), Aldrich chemical (Milwauk WI, including Sigma chemical and Fluka), apex chemical Ltd. (Miltonpark UK), Avocaado research (Lankaki, UK), BDHINC. (Toronto, Canada), Bionet (conway, UK), Chemservice Inc. (WestChester PA), Crescont chemical Co. (HauppaugenY), Eastmanorganic Chemicals, Eastmankodak company (Rochester, New York), Fisherscientific Co. (Pittsburgh chemical, PA), Fisonss Leicestershu, (Fronttierti scientific) (Lorentanut), icnbiomedia, Inc. (costamesa), KeyOrganics (conwatt, uk), lancaster synthesis (windhamnh), maybridge chemical co.ltd. (conwatt, uk), parshchemic co. (OremUT), Pfaltz & Bauer, Inc. (WaterburyCN), Polyorganix (houston, texas), pierce chemical co. (rockford, illinois usa), riedel haende haen ag (hannover, germany), spectrumq yield, Inc. (NewBrunswick, NJ), tciameria (portland, rusk), wornswald, Inc., and rockkommy chemical (richa).

Methods known to those skilled in the art can be identified by various reference books and databases. Suitable references and monographs detailing the synthetic methods of useful reactants for preparing the compounds described herein or articles giving references describing the preparation methods include, for example, "synthetic organic chemistry", john wiley & Sons, inc; sandleret, organic functional groups preambles, 2nd ed, academic press, new york, 1983; H.O.House, "modern synthetic reactions", 2nd Ed., W.A.Benjamin, Inc.Menlo park, Calif. Furia, 1972, T.L.Gilchrist, "heterocyclic chemistry", 2nd Ed., John Wiley & Sons, New York, 1992; march, "advanced organic chemistry: Reactions, mechanics and Structure", 4th ed., Wiley-Interscience, New York, 1992. Other suitable references and monographs detailing the synthesis of useful reactants for the preparation of the compounds described herein or giving reference to articles describing the preparation include, for example, Fuhrhop, J. and Penzlin, for "organic Synthesis: Concepts, Methods, Startingmaterials", Second, RevisedandEnlargededition (1994) John Wiley & SonsISBN: 3-527-; hoffman, R.V. "organic chemistry, Anmedia text" (1996) Oxford university Press, ISBN0-19-509618-5; Larock, R.C. "comprehensive organic transformations: AGuide functional group precursors" 2 d edition (1999) Wiley-VCH, ISBN:0-471 19031-4; March, J. "advanced organic chemistry: Reactions, Mechanism, and Structure" 4th edition (1992) John Wiley & Sons, ISBN:0-471 8712; Otera, J. "model chemistry" (2000) Wilkinson-VCH, ISBN: 0-1992; ISBN: 293-1992; ISBN: 27-824; ISBN: Islami-2000; ISBN: Ash-97-III.); solomons, T.W.G. "organic chemistry" 7th edition (2000) John Wiley & Sons, ISBN: 0-471-; "organic reactions" (1942-2000) John Wiley & Sons,55 volumes; and "chemistry of functional groups" John Wiley & Sons,73 volume.

Specific or similar reactants may also be identified by known chemical indexes provided by the american chemical society's chemical summary service (chemical abstractservice on american chemical society), which is available in most public and university libraries, and may also be available in online databases (more information is available at the american chemical society, washington, d.c.). Chemicals that are known but not sold in commercial catalogs may be prepared by a custom-made chemical organization, wherein many standard chemical custom-made organizations (e.g., those listed above) provide custom-made synthetic services. Reference is made to p.h.stahl & c.g.wermuth "handbook of pharmaceutical salts", verlaghelvetica chimica acta, Zurich,2002 for the preparation and selection of pharmaceutical salts of alkynylphenyl derivative compounds as described herein.

In general, the compounds disclosed herein can be prepared in a step-wise manner that includes the formation of alkynes and the formation of benzene ring side chains. Typically the formation of alkynes can be carried out by linking an alkyne precursor to a phenyl group. For example, in certain embodiments, an alkyne intermediate may be formed first, which forms a precursor to the alkynyl phenyl core structure. The side chain moiety, which is the linked precursor (i.e., propylene or ethylene oxide) and nitrogen-containing moiety of the compounds disclosed herein, can then be linked to the alkyne intermediate.

In other embodiments, the compounds disclosed herein can be prepared by the following steps: phenyl intermediates with appropriate side chains were first prepared, followed by alkyne formation to provide an alkynylphenyl core structure.

The following methods will illustrate various synthetic routes to alkyne intermediates and side chain moieties. One skilled in the art will recognize that methods of forming alkynes can be combined with methods of forming side chains to provide the compounds disclosed herein. For example, any of methods A-D can be combined with any of methods E-H, or any of methods I-J. They may also be further combined with any of the methods K-S to modify the linkages and/or the terminal nitrogen-containing moieties.

1. Formation of alkynes

The following methods A-D describe various methods of forming alkynes.

More specifically, method A illustrates the formation of the alkyne intermediate (A-3) in a Sonogashira or Castro-Stephens reaction. Depending on the order of reaction, Ar may be a phenyl derivative compound already attached to the side chain moiety, or Ar may comprise a reactive group (suitably protected) that will couple with the side chain moiety after the alkyne formation step.

According to method A, in the presence of a copper (I) catalyst (Castro-Stephens) or Pd⁰And Cu¹An alkyne (a-1) can be coupled to an aryl halide or reactive equivalent (a-2) in the presence of a mixture of catalysts (Sonogashira) to provide an alkyne intermediate (a-3).

Alkyne (A-1) toolHaving a terminal alkyne structure capable of coupling to A-2. The compounds containing different R can be prepared according to methods known in the art₅An alkyne of the group. For example, the organic halide can be converted to the corresponding alkyne (A-1) by coupling with acetylene. The halogenated benzenes or their reactive equivalents (A-2) may be commercially available or may be prepared by methods known in the art.

Palladium catalysts suitable for the coupling reaction are known to those skilled in the art. For example, exemplary palladium (0) catalysts include tetrakis (triphenylphosphine) palladium (0) [ Pd (PPh)₃)₄]And tetrakis (tris (o-tolylphosphine) palladium (0), tetrakis (dimethylphenylphosphine) palladium (0), tetrakis (tri-p-methoxyphenylphosphine) palladium (0), etc. it will be appreciated that palladium (II) salts may also be used which generate the palladium (0) catalyst in situ₂]Bis (triphenylphosphine) -palladium diacetate, and the like.

Copper catalysts suitable for the coupling reaction are well known to those skilled in the art. Typically, the copper (I) catalyst may be copper (I) iodide.

Method A

Method B shows the coupling of an organic halide (i.e., R)₅X) with a phenyl group (A-5) comprising a terminal alkynyl group optionally forms an alkyne intermediate (A-3).

Method B

Method C shows the formation of alkyne intermediate (A-7) by addition of a terminal alkyne group (A-5) to aldehyde or ketone (A-6).

Method C

Method D shows the formation of the alkyne intermediate (A-8) by addition of a terminal alkyne (A-5) to epoxide (A-9).

Method D

2. Side chain formation and modification

Methods E-S below describe various methods of side chain formation and modification.

In general, appropriately substituted phenyl derivatives can be coupled to a series of side chains, which can be further modified to provide the final attachment and nitrogen-containing moiety of the compounds disclosed herein.

Methods E-H illustrate the route by which the propylene linkage of the compounds disclosed herein is formed.

Method E illustrates the coupling of an aryl halide to allyl alcohol in the presence of a palladium (0) catalyst. The final alcohol group of allyl alcohol has been simultaneously oxidized to an aldehyde group, which can be further reductively aminated to an amine (-NR)₁₂R₁₃)。

Method E

Method F illustrates an aldol condensation reaction between an aryl aldehyde or ketone and a nitrile reagent containing at least one alpha-hydrogen. The resulting condensation intermediate may be further reduced to an amine (-NR) ₁₂R₁₃)。

Method F

Process G shows the acylation reaction to form a ketone-based linkage (i.e., R of formula (I))₁₀And R₁₁To form an oxo group). One skilled in the art will recognize that the R' group may contain functional groups that may be further modified.

Method G

Method H shows the ring-opening reaction of the epoxide reagent to form a hydroxyl-substituted propylene side chain linkage.

Method H

Method I illustrates the attachment of a side chain moiety, which may be an ethylene oxygen-linked precursor, through an oxygen. More specifically, by elimination of one molecule of H₂O, side chain precursor (R' OH) may be condensed with aryl derivatives. R' may include a functional group that may be further modified to prepare the attachment and nitrogen-containing moiety of formula (III) and its substructure including formula (IIIa) and (IIIb).

Method I

Method J shows a condensation reaction that provides an oxygen linking atom. Here, the condensation reaction results in elimination of the HX molecule.

Method J

After attachment, the side chain moieties may be further modified to provide the final attachment and terminal nitrogen-containing moieties of the compounds disclosed herein. The following methods illustrate various synthetic routes to utilize or modify side chain moieties by reduction, oxidation, nucleophilic or electrophilic substitution, fluorination, acylation, and the like. Thus, a variety of linkages can be synthesized.

Method K illustrates an amination process wherein a carboxylic acid is converted to an amine. Typically, the carboxylic acid (or ester) may be first reduced to a primary alcohol, which may then be converted to an amine by mesylate (mesylate), halide, azide, phthalimide, or Mitsunobu reactions, among others. For example, suitable reducing agents include sodium borohydride (NaBH)₄) Sodium cyanoborohydride (NaBH)₃CN), sodium triacetoxyborohydride (NaBH (OCOCH)₃)₃) Lithium aluminum hydride (LiAlH)₄) And the like. The resulting amine may be further functionalized by methods known in the art, as shown.

Method K

Additional or alternative modifications may be made in accordance with the methods set forth below.

Method L

Method M

Method N

Process O

Method P

Method Q

Process R

Method S

Equation I illustrates a complete synthetic method for preparing the compounds disclosed herein.

Reaction formula I

In reaction formula I, the side chain moiety is formed first, and the amino group is protected. The alkyne moiety is then formed by coupling with a terminal alkyne according to method A. The coupling product is then deprotected to give the final alkynylphenyl derivative terminated with a primary amine comprising a propylene linkage. Other nitrogen-containing moieties (-NR) according to methods known in the art₁₂R₁₃) May also be derived from terminal amines.

However, it will be appreciated by those skilled in the art that the order of the reactions may be varied. Thus, in other embodiments, the alkyne can be formed prior to attachment of the side chain.

Equation II illustrates a complete synthetic method for preparing the compounds disclosed herein.

Reaction formula II

In addition to the general reaction schemes and methods disclosed above, the present invention also provides other exemplary reaction schemes to illustrate methods of preparing any of the compounds disclosed herein.

Treatment of ophthalmic diseases and disorders

Alkynyl phenyl linked amine derivative compounds useful for treating ophthalmic diseases or disorders as described in detail herein, including compounds having a structure as set forth in any of formulae (a) - (G) and (I) - (III) and substructures thereof and specific alkynyl phenyl linked amine compounds as described herein, can inhibit one or more steps in the visual cycle, for example, by inhibiting or preventing trans-cis isomerase of the visual cycle, including the functional activity of trans-cis isomerase of the visual cycle (trans-cissomerase). The compounds described herein may inhibit, prevent, or in some way interfere with the isomerization step in the visual cycle. In certain embodiments, the compounds inhibit the isomerization of all-trans retinyl esters; in certain embodiments, the all-trans retinyl ester is a fatty acid ester of all-trans-retinol, and the compound inhibits isomerization of all-trans-retinol to 11-cis-retinol. The compound may adhere to or otherwise interact with an isomerase enzyme, also referred to herein and in the art as retinal isomerase or isomerohydrolase (isomerohydrolase), in some manner and inhibiting isomerase activity in at least one visual cycle. The compounds can prevent or inhibit the adhesion of an all-trans-retinyl ester substrate to an isomerase enzyme. Alternatively, or in addition, the compound may adhere to a catalytic site or region of an isomerase enzyme, thereby inhibiting the catalytic ability of the enzyme to isomerize an all-trans-retinyl ester substrate. Based on current scientific data, at least one isomerase that catalyzes the isomerization of all-trans-retinyl ester is believed to be located in the cytoplasm of RPE cells. As described herein, the various steps, enzymes, substrates, intermediates and products of the visual cycle have not been elucidated (see, e.g., Moiseyevetal, Proc. Natl. Acad. Sci. USA102:12413-18 (2004); Chenet al, Invest. Ophthalmol. Vis. Sci.47:1177-84 (2006); Lambert. supra).

As described herein and known in the art, methods for determining the effect of a compound on isomerase activity can be performed in vitro (Stecheret al, JBiolChem274:8577-85 (1999); see also Golczaketal, Proc. Natl. Acad. Sci. USA102:8162-67 (2005)). Retinal Pigment Epithelial (RPE) microsomal membranes isolated from animals (e.g., cattle, pigs, humans) can be used as the source of isomerase. The ability of amine derivative compounds having an alkynylphenyl group attached thereto to inhibit isomerase can also be tested by murine in vivo isomerase assays. It is known that brief exposure of the eye to intense light (either "photobleaching" or simply "bleaching" of the visual pigment) can photoisomerize nearly all of the 11-cis-retinal in the retina. The recovery of 11-cis-retinal after bleaching can be used to assess isomerase activity in vivo (see, e.g., Maedaetal., J.neurochem85:944-956(2003); VanHooseltal., JBiolChem277:19173-82, 2002). Electroretinographic (ERG) recording (electroretinographic (ERG) recording) as previously described (Haeselesereal, Nat. Neurosci.7:1079-87(2004); Sugitomoet, J. Toxicol.Sci.22suppl2:315-25(1997); Keratital, Documenta Ophthalmologica100:77-92(2000)) may be performed. See also De ignetetal, Science,244: 968-. In certain embodiments, the compounds for use in treating an individual having or at risk of having any of the ophthalmic and or retinal diseases or disorders described herein have IC₅₀Levels (concentration of compound at which 50% of isomerase activity is inhibited; as determined in the isomerase assay described herein or known in the art) are less than about 1 μ M; in other embodiments, the IC of the assay₅₀Levels of less than about 10 nM; in other embodiments, the IC of the assay₅₀Levels of less than about 50 nM; in certain other embodiments, the IC of the assay₅₀Levels of less than about 100 nM; in certain other embodiments, the IC of the assay₅₀Levels less than about 10 μ M; in other embodiments, the IC of the assay₅₀Levels less than about 50 μ M; in certain other embodiments, the IC of the assay₅₀Levels of less than about 100 μ M or about 500 μ M; in other embodiments, the IC of the assay₅₀The level is between about 1 μ M to 10 μ M; in other embodiments, the IC of the assay ₅₀Levels were between about 1nM and 10 nM. When administered to an individual, one or more compounds of the invention exhibit an ED50 value of about 5mg/kg, values below 5mg/kg being determined by inhibition of the isomerase reaction that leads to the production of 11-cis-retinol. In some embodiments, a compound of the invention has an ED50 value of about 1mg/kg when administered to an individual. In other embodiments, the compounds of the invention have an ED50 value of about 0.1mg/kg when administered to a subject. The ED50 value can be measured after administration of the compound or pharmaceutical composition thereof to a subject for about 2 hours, 4 hours, 6 hours, 8 hours, or more.

The compounds described herein are useful for treating an individual having an ophthalmic disease or disorder, particularly a retinal disease or disorder (e.g., age-related macular degeneration or stargardt's macular dystrophy). In one embodiment, the compounds described herein can inhibit (i.e., prevent, reduce, slow, eliminate, or minimize) the accumulation of lipofuscin pigment and lipofuscin-associated and/or related molecules in the eye. In another embodiment, the compounds can inhibit (i.e., prevent, reduce, slow, eliminate, or minimize) the accumulation of N-retinylidene-N-retinylethanolamine (A2E) in the eye. The ophthalmic disease may be due, at least in part, to accumulation of lipofuscin pigment and/or accumulation of A2E in the eye. Thus, in certain embodiments, methods of inhibiting or preventing the accumulation of lipofuscin pigment and/or A2E in the eye of an individual are provided. These methods comprise administering to the subject a composition comprising a pharmaceutically acceptable or suitable excipient (i.e., a pharmaceutically acceptable or suitable carrier) and an alkynylphenyl-linked amine derivative compound described in detail herein, including compounds having a structure described by any one of formulas (a) - (G) and (I) - (III) and substructures thereof, as well as the particular alkynylphenyl-linked amine compounds described herein.

Blinding retinal disease progression is associated with lipofuscin pigment accumulation in Retinal Pigment Epithelium (RPE) cells, including age-related macular degeneration (Delaeyeet, Retina15:399-406 (1995)). Lipofuscin microparticles are remnants of auto-fluorescent (autofluorescent) lysosomes (also known as age spots). The major fluorescent species of lipofuscin is A2E (orange-emitting fluorophore), a positively charged Schiff base condensation product of all-trans retinal and phosphatidylethanolamine (2:1 ratio) (see, e.g., Eldredal, Nature361:724-6 (1993); see also Sparrow, Proc. Natl. Acad. Sci. USA100:4353-54 (2003)). It is believed that many non-absorbed lipofuscin pigments originate from photoreceptor cells; deposition occurs in the RPE due to its entrapment in the membranous debris that photoreceptor cells eject daily. It is believed that the formation of this compound is not catalyzed by any enzyme, but rather A2E is formed by a spontaneous cycling reaction. In addition, A2E has a pyridine bis retinoid (pyridine bis retinoid) structure that is not degraded by enzymes once formed. Thus, lipofuscin and A2E accumulate as the human eye ages, and they also accumulate in the eyes of individuals with juvenile forms of macular degeneration known as stargardt disease (juvenileform) and several other congenital forms of retinal dystrophy.

A2E can cause retinal damage by different mechanisms. At low concentrations, A2E inhibited normal proteolysis in lysosomes (Holzer, invest. Ophthalmol. Vis. Sci.40:737-43 (1999)). At high concentrations, sufficient concentrations, A2E may act as a positively charged lysosomal detergent that lyses cell membranes and may alter lysosomal function, releasing apoptotic precursor proteins from mitochondria and ultimately destroying RPE cells (see, e.g., Eldredal et al, supra; Sparroweal, Invest, Ophthalmol. Vis. Sci.40:2988-95(1999); Holzel. Supra; Finnemadetal, Proc. Natl. Acad. Sci.USA99:3842-347(2002); Suteret al, J.biol. chem.275:39625-30 (2000)). A2E is phototoxic and causes blue light-induced RPE cell apoptosis (see, e.g., sparrowet, invest, ophthalmol, vis. sci.43:1222-27 (2002)). Photooxidation products (e.g., epoxides) of A2E that damage cellular macromolecules, including DNA, form when exposed to blue light (sparrowwet, j.biol. chem.278(20):18207-13 (2003)). The self-generated singlet oxygen of A2E reacts with A2E to form an epoxide at the carbon-carbon double bond (sparrowwet et al, supra). The generation of oxygen reactive species by photoexcited A2E leads to oxidative damage to the cells, which often leads to cell death. Indirect methods to prevent the formation of A2E by inhibiting the biosynthesis of the direct precursor all-trans-retinal of A2E have been described (see U.S. patent application publication No. 2003/0032078). However, the usefulness of the methods described therein is limited because the production of all-trans retinal is an important component of the visual cycle. Other therapies described include neutralization of damage caused by oxidative species by using superoxide dismutase mimicry (see, e.g., U.S. patent application publication No. 2004/0116403) and inhibition of cytochrome C oxidase caused by A2E in retinal cells with negatively charged phospholipids (see, e.g., U.S. patent application publication No. 2003/0050283).

The alkynylphenyl-linked amine derivative compounds described herein may be used to prevent, reduce, inhibit, or reduce the accumulation (i.e., deposition) of A2E and A2E-related and/or derivatized molecules in RPE. Without being limited by theory, since RPE is critical to maintaining photoreceptor cell integrity, preventing, reducing, or inhibiting damage to RPE can inhibit degeneration of retinal neurons, particularly photoreceptor cells (i.e., enhance cell viability or increase or prolong cell viability). Compounds that specifically bind or interact with A2E, A2E-related and/or derived molecules or affect A2E formation and accumulation may also reduce, inhibit, prevent or reduce the toxic effects of one or more A2E, A2E-related and/or derived molecules that cause damage, death or neurodegeneration of retinal nerve cells (including photoreceptor cells), or in some way reduce retinal nerve cell viability. These toxic effects include induction of apoptosis, self-production of singlet oxygen, and production of oxygen reactive species; the self-production of singlet oxygen forms A2E-epoxide, which causes DNA damage, thereby damaging cellular DNA and causing cellular damage; lysing the cell membrane; altering lysosomal function; and affecting the release of apoptotic precursor protein from mitochondria.

In other embodiments, the compounds described herein may be used to treat other ophthalmic diseases or disorders, for example, glaucoma, cone-rod dystrophy, retinal detachment, hemorrhagic or hypertensive retinopathy, retinitis pigmentosa, optic neuropathy, inflammatory retinopathy, proliferative vitreoretinopathy, congenital retinal dystrophy, damage to the optic nerve (e.g., physical damage, overexposure damage, or laser), inherited optic neuropathy, neuropathy due to toxic agents or due to adverse drug reactions or vitamin deficiencies, Sorsby's fundus dystrophy, uveitis, retinal disorders associated with alzheimer's disease; retinal disorders associated with multiple sclerosis, retinal disorders associated with viral infections (cytomegalovirus or herpes simplex virus), retinal disorders associated with parkinson's disease, retinal disorders associated with AIDS, or other forms of progressive retinal atrophy or degeneration. In other specific embodiments, the disease or disorder is caused by mechanical injury, chemical or drug-induced injury, thermal injury, radiation injury, photodamage, laser injury. The subject compounds are useful for treating genetic or non-genetic retinal dystrophies. These methods are also useful for preventing ocular damage due to environmental factors such as light-induced oxidative retinal damage, laser-induced retinal damage, "light bomb damage" or "light glare" including, but not limited to, myopic ametropia (see, e.g., Quinn genetic, Nature1999;399: 113-.

In other embodiments, provided herein is the use of any one or more of the alkynyl-phenyl-linked amine-derived compounds described in detail herein, including compounds having a structure according to any one of formulas (a) - (G) and (I) - (III) and substructures thereof, as well as specific alkynyl-phenyl-linked amine compounds described herein, to inhibit the formation of neovasculature in the retina, including but not limited to neovascular glaucoma. In certain other embodiments, methods of reducing retinal hypoxia using the compounds described herein are provided. These methods include those in the actual need for administration to an individual, the administered pharmaceutical composition comprising a pharmaceutically acceptable or suitable excipient (i.e., a pharmaceutically acceptable or suitable carrier) and an alkynyl-phenyl linked amine derivative compound described in detail herein, including compounds having a structure described by any one of formulas (a) - (G) and (I) - (III) and substructures thereof, as well as the specific alkynyl-phenyl linked amine compounds described herein.

By way of explanation only, and not limitation to any theory, and as described in further detail herein, dark-adapted rod photoreceptors result in very high metabolic demands (i.e., energy expenditure (ATP expenditure) and oxygen expenditure). The resulting hypoxia may cause and/or exacerbate retinal degeneration that may worsen in cases where the retinal vasculature has undergone damage, including, but not limited to, for example, diabetic retinopathy, macular edema, diabetic maculopathy, retinal vessel occlusion (which includes retinal vein occlusion and retinal artery occlusion), retinopathy of prematurity, ischemia reperfusion-related retinal damage, and wet age-related macular degeneration (AMD). In addition, retinal degeneration and hypoxia can lead to the formation of new blood vessels, which in turn can exacerbate the degree of retinal degeneration. The alkynyl phenyl linked amine derivative compounds described herein that modulate visual cycle can be administered to prevent, inhibit and/or delay dark adaptation of rod photoreceptor cells, and can thus reduce metabolic demand, thus reducing hypoxia and inhibiting neovascularization.

In this context, oxygen is a key molecule for maintaining retinal function in mammals, and retinal hypoxia may be a factor in many retinal diseases and disorders with ischemic requirements. In most mammals, including humans, having a two-dimensional tube coupling supplying the retina, oxygenation of the interior of the retina is achieved by the microvasculature within the retina, which is sparse compared to the choriocapillaris supplying RPE and photoreceptors with oxygen. Different vascular connections supply networks to create uneven oxygen tension across the thickness of the retina (cringletal, invest. ophthalmol. vis. sci.43:1922-27 (2002)). Fluctuations in oxygen across the retinal layers are associated with different capillary densities and differences in oxygen consumption by various retinal neurons and glia.

Local oxygen tension can significantly affect the retina and its microvasculature by modulating a range of vasoactive substances including, for example, Vascular Endothelial Growth Factor (VEGF). (see, e.g., Werdichetal, Exp. EyeRes.79:623(2004); ardenetal, Br. J. Ophthalmol.89:764 (2005)). Rod photoreceptors are believed to have the highest metabolic rate of any cell in the body (see, e.g., ardenetal, supra). During dark adaptation, the rod photoreceptors restore their high cytoplasmic calcium concentrations by having cGMP-gated calcium channels with concomitant extrusion of sodium ions and water. In contrast to photoadaptive (i.e., photopic) conditions, sodium efflux from cells is an ATP-dependent process, resulting in approximately 5-fold more oxygen consumption by retinal neurons in dark-adapted (i.e., dark-adapted) conditions. Thus, during typical dark adaptation of the photoreceptor, high metabolic demand in the dark adapted retina leads to a locally significant decrease in oxygen concentration (ahmedetal, invest. ophthalmol. vis. sci.34:516 (1993)).

Without being bound by any theory, retinal hypoxia is further exacerbated in the retina of individuals suffering from a disease or disorder (e.g., central retinal vein occlusion, where the retinal vasculature has been damaged). The aggravation of hypoxia causes the easy influence on vision and the formation of retinal neovascularization. Neovascularization is the formation of a new, functional network of microvessels in which red blood cells perfuse, and neovascularization is a typical retinal degenerative disorder including, but not limited to, diabetic retinopathy, retinopathy of prematurity, wet AMD and central retinal vein occlusion. Preventing or inhibiting dark adaptation of rod photoreceptor cells, thereby reducing energy and oxygen consumption (i.e., reducing metabolic demand), may inhibit or slow retinal degeneration, and/or may promote regeneration of retinal cells including rod photoreceptor cells and Retinal Pigment Epithelium (RPE) cells, and may reduce hypoxia and may inhibit neovascularization.

Described herein are methods of inhibiting (i.e., reducing, preventing, slowing, or delaying in a biologically or statistically significant manner) degeneration of retinal cells, including retinal nerve cells and RPE cells described herein, and/or reducing (i.e., preventing or slowing, inhibiting, eliminating in a biologically or statistically significant manner) retinal ischemia. Also provided herein are methods of inhibiting (i.e., reducing, preventing, slowing, or delaying in a biologically or statistically significant manner) the formation of new blood vessels in an eye, particularly the retina. These methods comprise contacting the retina, and thus the retinal cells (including retinal nerve cells, such as rod photoreceptor cells and RPE cells), with at least one alkynylphenyl-linked amine-derived compound described herein that inhibits trans-cis isomerase (which may include inhibiting isomerization of all-trans-retinyl ester) for at least one visual cycle under conditions and for a time that can prevent, inhibit or delay dark adaptation of the rod photoreceptor cells in the retina. As described in further detail herein, in particular embodiments, the compounds that contact the retina interact with isomerase or enzyme complex (enzymatocccomplex) of RPE cells in the retina and inhibit, block, or somehow interfere with the catalytic activity of the isomerase. Thus, isomerization of all-trans-retinyl esters is inhibited or reduced. At least one alkynylphenyl-linked amine derivative compound (or composition comprising at least one compound) can be administered to individuals who have developed and developed an ophthalmic disease or disorder, or those at risk of developing an ophthalmic disease or disorder, or individuals who exhibit or are at risk of developing, for example, a neovascular or retinal ischemic disorder.

In this context, the visual cycle (also called the retinoid cycle) refers to the enzyme series and to the light-mediated conversion that occurs between the 11-cis and all-trans forms of retinol/retinal in the photoreceptors and Retinal Pigment Epithelium (RPE) cells in the eye. In vertebrate photoreceptors, photons cause isomerization of the 11-cis-retinylidene chromophore to the all-trans-retinylidene paired with the visual opsin receptor. This photoisomerization causes a conformational change in the opsin protein which, in turn, triggers a biological chain reaction called light transduction (filipek., annu. rev. physiol.65851-79 (2003)). Regeneration of the visual chromophore following absorption of light and photoisomerization of 11-cis-retinal to all-trans retinal is a critical step in restoring the photoreceptor to its dark-adapted state. Regeneration of the opsin requires reversion of the chromophore to the 11-cis-conformation (see McBeeetal, prog. RetineEyeRes.20: 469-52 (2001)). The chromophore is released from the opsin and reduced by retinol dehydrogenase in the photoreceptor. The product, all-trans-retinol, is entrapped in the adjacent Retinal Pigment Epithelium (RPE) in the form of insoluble fatty acid esters with the subcellular structure of the known retinal body (imanishietal, j.cellbiol.164:373-78 (2004)).

In the visual cycle of rod-shaped receptor cells, the 11-cis-retinal chromophore (called rhodopsin) within the retinoid molecule absorbs photons of light and isomerizes into an all-trans configuration, thus activating a light transduction cascade. Rhodopsin is a G protein-coupled receptor (GPCR) that consists of seven transmembrane helices interconnected by extracellular and cytoplasmic loops. When the all-trans form of the retinoid remains covalently attached to the pigment molecule, the pigment is called a rhodopsin, which exists in different forms (e.g., rhodopsin I and rhodopsin II). The all-trans retinoid is then hydrolyzed and the opsin is in the form of a apo-protein, opsin, which is also known in the art and herein as apo-rhodopsin. This all-trans retinoid is transported or accompanies away from the photoreceptor cells and passes through the extracellular space into the RPE cells, where the retinoid is converted to the 11-cis isomer. The movement of retinoids between RPE and photoreceptor cells is thought to be achieved by different chaperone polypeptides in various cell types. See Lambert, progressive Retinalkand eye research23:307-80 (2004).

Rhodopsin undergoes a continuous transition between the three forms (rhodopsin, metamorphic rhodopsin and apo rhodopsin) in the presence of light. Rod photoreceptors are in a "dark-adapted" state when most of the visual pigment is in the form of rhodopsin (i.e., linked to 11-cis retinal). The state of the photoreceptor cell is called "photoadaptation" when the opsin is mostly in the form of a modified rhodopsin (i.e., linked to all-trans retinal), and "rhodopsin-absent" when the opsin is an aporhodopsin (or opsin) and there is no tethered chromophore. The three states of photoreceptor cells each have different energy requirements and consume different amounts of ATP and oxygen. In the dark adaptation state, the rhodopsin has no modulating effect on the open cation channels, resulting in cations (Na) ⁺/K⁺And Ca²⁺) Into the flow of (a). In order to maintain these cations at appropriate concentrations in the cell in the dark state, the photoreceptor cells actively transport the cations out of the cell by an ATP-dependent pump (ATP-dependent pump). Thus maintaining this "Dark current "requires a large amount of energy, resulting in high metabolic demand. In the photoadaptive state, rhodopsin causes an enzyme cascade process leading to GMP hydrolysis, which in turn closes the cation specific channels in the photoreceptor cell membrane. In the rhodopsin deficient state, the chromophore is hydrolyzed from the rhodopsin to form apoproteins, opsins (apolacyl rhodopsin), which partially regulate the cation channel, so that rod photoreceptor cells display a reduced current compared to photoreceptors in the dark-adapted state, resulting in a reduced need for proper metabolism.

Under normal light conditions, the occurrence of rod photoreceptors in the dark adaptation state is low, typically below 2%, and the cell is predominantly in the light-adapted or rhodopsin deficient state, which will generally result in a relatively low metabolic demand compared to cells in the dark adaptation state. However, at night, there is no photoadaptation and since the "dark" visual cycle in RPE cells works continuously, the rod photoreceptor cells will be supplemented with 11-cis-retinal, and the relative incidence of the appearance of dark-adapted photoreceptor states increases dramatically. This shift in dark adaptation of rod photoreceptors leads to increased metabolic demand (i.e., increased ATP and oxygen consumption), ultimately leading to retinal hypoxia and its subsequent initiation of angiogenesis. Thus, most ischemic injury occurs in a dark state, such as when sleeping at night.

Without being bound to any theory, medical intervention during the "dark" visual cycle may prevent retinal hypoxia and the formation of new blood vessels triggered by the high metabolic activity of rod photoreceptor cells under dark adaptation. By way of example only, by altering the "dark" visual cycle through the administration of any of the compounds described herein, which are isomerase inhibitors, rhodopsin (i.e., 11-cis retinal binding) can be reduced or eliminated, thereby preventing or inhibiting dark adaptation of rod photoreceptors. In turn, may reduce retinal metabolic demand, reduce the risk of nocturnal retinal ischemia and neovascularization, and thus inhibit or slow retinal degeneration.

In one embodiment, at least one of the compounds described herein (i.e., the alkynyl-phenyl-linked amine-derived compounds described in detail herein, including compounds having a structure described by any of formulae (A) - (G) and (I) - (III) and substructures thereof, and the specific alkynyl-phenyl-linked amine compounds described herein) prevents, slows, inhibits, or in some way attenuates the catalytic activity of visual cycle isomerase in a statistically or biologically significant manner, it can prevent, inhibit, or delay dark adaptation of rod photoreceptor cells, thereby inhibiting (i.e., slowing, eliminating, preventing, delaying progression of, or reducing in a statistically or biologically significant manner) degeneration of (or increasing the survival of) retinal cells of the retina in the eye. In another embodiment, the alkynyl phenyl linked amine derivative compound can prevent or inhibit dark adaptation of rod photoreceptor cells, thereby reducing ischemia (i.e., statistically or biologically in a significant manner reducing, preventing, inhibiting, slowing the progression of ischemia). In yet another embodiment, any of the alkynyl phenyl-linked amine-derivative compounds described herein can prevent dark adaptation of rod photoreceptor cells, thereby inhibiting neovascularization of the retina in the eye. Accordingly, provided herein are methods of inhibiting retinal cell degeneration, inhibiting retinal neovascularization in the eye of an individual, and reducing ischemia in the eye of an individual under conditions and for a time sufficient to prevent, inhibit, or delay dark adaptation of rod photoreceptors, wherein the methods comprise administering at least one alkynyl phenyl linked amine compound as described herein. These methods and compositions are therefore useful for treating ophthalmic diseases or disorders, including but not limited to diabetic macular degeneration, diabetic maculopathy, retinal vessel occlusion, retinopathy of prematurity, or ischemia reperfusion-related retinal damage.

The alkynylphenyl amine derivative compounds described herein (i.e., the alkynylphenyl-linked amine derivative compounds described in detail herein, including compounds having a structure described by any of formulas (a) - (G) and (I) - (III) and substructures thereof, as well as the specific alkynylphenyl-linked amine compounds described herein) can prevent (i.e., retard, slow, inhibit, or reduce) the recovery of the retinoid chromophore, can prevent or inhibit or delay the formation of retinal, and can increase the concentration of retinyl esters, which perturb the visual cycle, inhibit the regeneration of rhodopsin, and can prevent, slow, delay, or inhibit dark adaptation of rod photoreceptor cells. In certain embodiments, when dark adaptation of rod photoreceptor cells is prevented in the presence of the compound, dark adaptation is substantially prevented and the number or percentage of rod photoreceptor cells lacking rhodopsin or photoadaptive in the absence of the drug is increased as compared to the number or percentage of rod photoreceptor cells lacking rhodopsin or photoadaptive in the absence of the drug. Thus, in certain embodiments, when dark adaptation of rod photoreceptor cells is prevented (i.e., substantially prevented), only at least 2% of the rod photoreceptor cells are dark adapted, similar to the number or percentage of cells in the dark adapted state under normal lighting conditions. In other embodiments, at least 5-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, or 60-70% of the rod photoreceptor cells are dark-adapted after administration of the drug. In other embodiments, the compound acts to delay dark adaptation, and dark adaptation of the rod photoreceptors in the presence of the compound may be delayed by 30 minutes, 1 hour, 2 hours, 3 hours, or 4 hours, as compared to dark adaptation of rod photoreceptors in the absence of the compound. By contrast, when an alkynylphenylamine derivative compound is administered such that the compound effectively inhibits isomerization of the substrate under photoadaptive conditions, the compound is administered in a manner that minimizes the percentage of dark-adapted rod photoreceptors, e.g., only 2%, 5%, 10%, 20%, or 25% of the rod photoreceptors are dark-adapted (see, e.g., U.S. patent application publication No. 2006/0069078; patent application No. PCT/US 2007/002330).

In the retina in the presence of the at least one alkynylphenylamine derivative compound attached, regeneration of rhodopsin in the rod photoreceptor cells is inhibited or the rate of regeneration is reduced (i.e., inhibited, reduced or decreased in a statistically or biologically significant manner), at least in part, by preventing the formation of retinal, reducing the concentration of retinal, and/or increasing the concentration of retinyl ester. To determine the concentration of rhodopsin regeneration by rod photoreceptors, the concentration of rhodopsin regeneration (which is also referred to as the first concentration) can be determined before the compound is brought into contact with the retina (i.e., before administration of the drug). After a sufficient period of time for the compound to interact with the retina and cells in the retina has elapsed (i.e., after administration of the compound), the concentration of rhodopsin regeneration (which may be referred to as a second concentration) may be determined. A decrease in the second concentration compared to the first concentration indicates that the compound inhibits rhodopsin regeneration. The concentration of rhodopsin production can be measured after each administration or after any number of administrations, with progress through the treatment regimen being manifested as the effect of the drug in inhibiting rhodopsin regeneration.

In certain embodiments, the subject in need of treatment described herein may have a disease or disorder that causes or contributes to a reduction in the ability of rod photoreceptors in the retina to regenerate rhodopsin. For example, inhibition of rhodopsin regeneration (reducing the rate of rhodopsin regeneration) may be indicative of an individual with diabetes. In addition to determining the rhodopsin regeneration concentration of diabetic individuals before and after administration of the alkynylphenylamine derivative compounds described herein attached, the effect of the compounds can also be shown by comparison as follows: comparing the inhibitory effect of rhodopsin regeneration in a first individual (or a first group or plurality of individuals) to a second individual (or a second group or plurality of individuals) having diabetes but not administered the drug.

In another embodiment, a method of preventing or inhibiting rod photoreceptor cell(s) dark adaptation of a retina is provided, comprising contacting the retina with at least one alkynyl-phenyl-linked amine derivative compound described herein (i.e., an alkynyl-phenyl-linked amine derivative compound described in detail herein, including compounds having a structure according to any one of formulae (a) - (G) and (I) - (III) and substructures thereof, and a particular alkynyl-phenyl-linked amine compound described herein) under conditions and for a time sufficient for the drug to interact with an isomerase enzyme in the retinal cell (e.g., RPE cell). A first concentration of 11-cis-retinal in the rod photoreceptor cell in the presence of the compound can be determined and compared to a second concentration of 11-cis-retinal in the rod photoreceptor cell in the absence of the compound. When the first concentration of 11-cis-retinal is less than the second concentration of 11-cis-retinal, it is indicated that dark adaptation of rod photoreceptor cells is prevented or inhibited.

Inhibiting the regeneration of rhodopsin may also comprise increasing the concentration of 11-cis-retinyl ester present in RPE cells in the presence of the compound compared to the concentration of 11-cis-retinyl ester present in RPE cells in the absence of the compound (i.e., prior to administration of the drug). The retinal structure of RPE, which is believed to be used to store retinyl esters, can be observed and analyzed using two-photon imaging techniques (two-photon imaging techniques) (see, e.g., imanishietal, j. cellbiol.164:373-83(2004), Epub2004January 26). A first concentration of retinyl ester can be measured prior to administration of the compound, and a second concentration of retinyl ester can be measured after the first administration or any subsequent administration, wherein an increase in the second concentration compared to the first concentration indicates that the compound inhibits regeneration of rhodopsin.

Retinyl esters were analyzed by gradient HPLC according to methods commonly used in the art (see, e.g., Mataeal., Neuron36:69-80(2002); Trevinoal.J.Exp.biol.208: 4151-57 (2005)). For the measurement of 11-cis and all-trans retinal, the extraction of retinoids by formaldehyde (see, e.g., Suzukietal, Vis. Res.28:1061-70 (1988); OkajimaandPepperberg, Exp. Eye Res.65:331-40(1997)) or by hydroxylamine (see, e.g., Groennedijkey, Biochim. Biophys. acta.617:430-38(1980)) can be used prior to isocratic HPLC (see, e.g., Trevinoetal, supra) analysis. Retinoids can be detected using a spectrophotometer (see, e.g., Maedaetal., J.neurochem.85:944-956(2003); VanHooseltal., J.biol.chem.277:19173-82 (2002)).

Embodiments of the methods described herein for treating ophthalmic diseases or disorders, inhibiting retinal cell degeneration (or increasing retinal cell survival), inhibiting neovascularization, and reducing retinal ischemia, preventing or inhibiting dark adaptation of rod photoreceptors in the retina include increasing the concentration of apophotopurpurin (also known as opsin) in the photoreceptors. The total concentration of visual pigment is close to the total amount of rhodopsin and apophotopurpurin and the total concentration is kept constant. Thus, preventing, retarding, or inhibiting dark adaptation of rod photoreceptor cells may alter the ratio of apophotopurpurin to rhodopsin. In particular embodiments, preventing, retarding, or inhibiting dark adaptation by administering the alkynyl phenyl linked amine derivative compounds described herein may increase the ratio of aporhodopsin concentration to rhodopsin concentration compared to the ratio without administration of the drug (e.g., prior to administration of the drug). An increase in the ratio of apophotoviologen to rhodopsin (i.e., a statistically or biologically significant increase) indicates an increase in the percentage or number of rhodopsin-deficient rod photoreceptors and a decrease in the percentage or number of dark-adapted rod photoreceptors. The effect of the drug can be measured by measuring the ratio of apoplectic rhodopsin to rhodopsin throughout the course of treatment.

The determination and characterization of the ability of a compound to prevent, delay or inhibit rod photoreceptor dark adaptation can be determined from animal model studies. The concentration of rhodopsin and the ratio of aporhodopsin to rhodopsin (which may be referred to as the first concentration or first ratio, respectively) may be determined prior to administration of the drug, and then the concentration of rhodopsin and the ratio of aporhodopsin to rhodopsin (which may be referred to as the second concentration or second ratio, respectively) may be determined after the first or any subsequent administration to determine or demonstrate that the concentration of aporhodopsin is greater than the concentration of aporhodopsin in the retina of the animal to which the drug is not administered. The concentration of rhodopsin in rod photoreceptors can be determined according to methods commonly used in the art and provided herein (see, e.g., yanetal.j. biol. chem.279: 48189-96 (2004)).

The subject in need of such treatment may be a human or may be a non-human primate or other animal (i.e., veterinary use), who has developed symptoms of, or is at risk of developing, an ophthalmic disease or disorder. Examples of non-human primates and other animals include, but are not limited to, farm animals, pets, and zoo animals (e.g., horses, cows, buffalos, llamas, goats, rabbits, cats, dogs, chimpanzees, orangutans, gorillas, monkeys, elephants, bears, large felines, and the like).

Also provided herein are methods of inhibiting (reducing, slowing, preventing) degeneration and increasing the survival rate (or prolonging cell viability) of retinal nerve cells comprising administering to an individual a composition comprising a pharmaceutically acceptable carrier and an alkynylphenyl-linked amine derivative compound described herein, including compounds having a structure described by any one of formulae (a) - (G) and (I) - (III) and substructures thereof, and the specific alkynylphenyl-linked amine compounds recited herein. Retinal nerve cells include photoreceptor cells, bipolar cells, horizontal cells, ganglion cells, and amacrine cells. In another embodiment, methods are provided for increasing survival or inhibiting degeneration of developing mature retinal cells, such as RPE cells and mullerian cells. In other embodiments, methods of preventing or inhibiting degeneration of photoreceptors of an eye of an individual are provided. Methods of preventing or inhibiting degeneration of photoreceptors in an eye of an individual include methods of restoring photoreceptor function in an eye of an individual. Such methods comprise administering to the subject a composition comprising an alkynylphenyl linked amine derivative compound as described herein and a pharmaceutically acceptable carrier (e.g., excipient or adjuvant). More specifically, the methods comprise administering to the subject a pharmaceutically acceptable excipient and an alkynylphenyl linked amine derivative compound described herein, including compounds having any one of the structures described in formulas (a) - (G) and (I) - (III) or substructures thereof described herein. Without being bound by theory, the compounds described herein may inhibit the isomerization step of the retinoid cycle (i.e., the visual cycle) in the eye and/or may slow the amount of chromophores that retinoid cycle.

Ophthalmic diseases are caused, at least in part, by the accumulation of lipofuscin pigment in the eye and/or by the accumulation of N-retinylidene-N-retinylethanolamine (A2E). Thus, in certain embodiments, methods of inhibiting or preventing the accumulation of lipofuscin pigment and/or A2E in the eye of an individual are provided. These methods comprise administering to the subject a pharmaceutically acceptable carrier and a composition of an alkynylphenyl linked amine compound described in detail herein, including compounds having the structures described in general formulas (a) - (G) and (I) - (III), or substructures thereof.

The amine compound having an alkynylphenyl group attached thereto can be administered to an individual who has an excess amount of retinoid in the eye (e.g., an excess of 11-cis-retinol or 11-cis-retinal), an excess of retinoid waste or intermediates in the all-trans-retinal cycle, or the like. During or after administration of any of the compounds described herein, the individual can be assayed for alterations (increases or decreases in a statistically or physiologically significant manner) in one or more endogenous retinoids using the methods described herein and as are commonly used in the art. Rhodopsin, which consists of the proteins opsin and retinal (forms of vitamin a), is localized in the cell membrane of the photoreceptor cells of the eye and catalyzes the only photosensitizing step in vision. The 11-cis-retinal chromophore is located in the protein's capsule and isomerizes to all-trans retinal upon absorption of light. Isomerization of retinal leads to a change in the shape of rhodopsin, which elicits a response that results in a cascade of nerve impulses transmitted through the optic nerve to the brain.

For example, in U.S. patent application publication No. 2005/0159662 (the disclosure of which is incorporated herein by reference in its entirety), a method for determining the concentration of a retinoid endogenous to the eye of a vertebrate and the excess or deficiency of such retinoid is disclosed. Other methods of determining the concentration of an endogenous retinoid in an individual that are useful for measuring whether such concentration of the retinoid is above a normal range include, for example, High Pressure Liquid Chromatography (HPLC) analysis of the retinoid from a biological sample of the individual. Retinoid concentrations can be determined, for example, from biological samples (blood samples, including serum and plasma) of an individual. Biological samples may also include vitreous humor, aqueous humor, intraocular fluid, subretinal fluid, or tears.

For example, a blood sample can be obtained from an individual and the concentration of different retinoids and one or more retinoids in the sample can be separated and analyzed by normal phase High Pressure Liquid Chromatography (HPLC) (e.g., using HP1100HPLC and a Beckman, Ultrasphere-Si,4.6mm X250 mm column, using 10% ethyl acetate/90% hexane at a flow rate of 1.4 ml/min). For example, retinoids can be detected at 325nm using a diode array detector and HPChemstation.03.03 software. For example, excess retinoid may be determined by comparing the characteristics (i.e., qualitatively, e.g., the identity of a particular compound, and quantitatively, e.g., the respective concentrations of a particular compound) of the retinoid in a sample with a sample from a normal human. Those skilled in the art familiar with such detection and techniques will readily understand the inclusion of appropriate controls.

As described herein, an increased or excess amount of an endogenous retinoid (e.g., 11-cis-retinol or 11-cis-retinal) refers to a concentration of the endogenous retinoid that is higher than the concentration detected in healthy eyes of young vertebrates of the same population. Administration of an alkynylphenyl-linked amine derivative compound and reduction or elimination of the need for endogenous retinoids. In certain embodiments, the concentration of an endogenous retinoid in an individual can be compared before or after any one or more doses of an alkynylphenyl linked amine compound are administered to the individual to determine the effect of the compound on the endogenous retinoid concentration in the individual.

In another embodiment, the methods of treating an ophthalmic disease or disorder, inhibiting neovascularization, and reducing ischemia of the retina described herein comprise administering at least one alkynyl phenyl-linked amine compound described herein, thereby resulting in a reduction in metabolic demand, which comprises resulting in a reduction in ATP and oxygen consumption in rod photoreceptor cells. As described herein, ATP and oxygen consumption of dark-adapted rod photoreceptors is greater than ATP and oxygen consumption of photoadaptive or rhodopsin-deficient rod photoreceptors; thus, use of the compound in the described methods can prevent, inhibit or delay dark adaptation and reduce ATP depletion of rod photoreceptor cells compared to dark adapted rod photoreceptor cells (e.g., cells prior to administration of the compound or contact with the compound or those that have not been exposed to the compound).

The methods described herein that can prevent or inhibit rod photoreceptor dark adaptation can thus reduce hypoxia (i.e., decrease in a statistically or biologically significant manner) of the retina. For example, the level of hypoxia (first level) can be determined prior to initiation of treatment, that is, prior to the first administration of the compound (or composition comprising the compound, as described herein). The grade of hypoxia (e.g., the second grade) is determined after the first administration and/or any second or subsequent administration to detect or characterize hypoxia throughout the treatment regimen. A reduction (decrease) in the second (or any subsequent) level of hypoxia compared to the level of hypoxia prior to the beginning of administration indicates that the compounds and treatment regimens prevent dark adaptation of rod photoreceptor cells and can be used to treat ophthalmic diseases and disorders. Oxygen consumption, oxygenation of the retina, and/or hypoxia of the retina can be measured using methods commonly used in the art. For example, oxygenation of the retina can be determined by measuring fluorescence of flavoproteins of the retina (see U.S. patent No. 4,569,354). Another exemplary method is retinal oximetry, which is the measurement of oxygen saturation of blood in the large blood vessels of the retina near the optic disc. This method can be used to confirm and determine the extent of retinal hypoxia prior to changes in retinal vasculature that can be detected.

The biological sample may be a blood sample (from which serum and plasma may be prepared), a biopsy specimen, a bodily fluid (e.g., vitreous fluid, aqueous humor, ocular fluid, subretinal fluid, or tears), a tissue explant (tissue explant), an organ culture, or any other tissue or cell preparation obtained from an individual or other biological source. The sample may further relate to a tissue or cell preparation in which the intact morphology or physical state has been disrupted by, for example, dissection, separation, solubilization, fractionation, homogenization, biochemical or chemical extraction, comminution, lyophilization, sonication, or other methods for processing samples derived from an individual or biological source. The individual or biological source may be a human or non-human animal, a primary cell culture (e.g., a retinal cell culture), or a cultured cell line, including, but not limited to, a genetic cell line, an immortalized or immortalized cell line, a somatic hybrid cell line, a differentiated or differentiable cell line (differentiated cells), a variant cell, and the like, that may comprise chromosomally integrated or episomal recombinant nucleic acid sequences. As described herein, mature retinal cells including retinal nerve cells, RPE cells, and Muller cells (Mullerglialcells) can be present in or isolated from a biological sample. For example, mature retinal cells can be obtained from primary or long-term cell cultures, or can be present in a biological sample obtained from an individual (human or non-human animal).

3. Retinal cells

The retina is a thin layer of nerve tissue, located between the vitreous and choroid in the eye. The major landmarks in the retina are the fovea, macula, and optic disc. The retina is thickest near the posterior pole zone and thins near the periphery. The spot is located on the posterior portion of the retina, including the fovea and the fovea. The fovea contains the area of greatest cone cell density and therefore has the sharpest vision on the retina. The fovea is contained within the fovea, which is the macula.

The peripheral portion of the retina increases the field of vision. The peripheral retina extends anteriorly along the eye's ciliary body and is divided into four regions: a proximal perimeter (rearmost), a middle perimeter, a distal perimeter, and a serrated edge (foremost). Jagged edge refers to the end of the retina.

The term neuron (or nerve cell) as understood in the art and used herein means a cell produced from a neuroepithelial cell precursor. Mature neurons (i.e., fully differentiated cells) display several specific antigenic markers. Neurons can be functionally divided into three groups: (1) afferent neurons (or sensory neurons) that transmit information to the brain to recognize the sensory and motor coordination portions; (2) motor neurons that deliver instructions to muscles and glands; and (3) interneurons, which are responsible for local loops; and (4) projection interneurons, which transfer information from one region of the brain to another, and thus have long axons. Interneurons process information in specific sub-regions of the brain and have relatively short axons. A neuron typically has four defined regions: cell body (soma); an axon; a dendrite; and presynaptic terminals. Dendrites are primarily responsible for the transmission of information from other nerve cells. Axons carry electrical signals that originate from the cell body to other neurons or effector organs. At the presynaptic terminal, the neuron transmits information to another cell (post-synaptic cell), which may be another neuron, a muscle cell or a secretory cell.

The retina is composed of several types of nerve cells. As described herein, the types of retinal nerve cells that can be cultured in vitro using this method include photoreceptor cells, ganglion cells, and interneurons, e.g., bipolar cells, horizontal cells, and amacrine cells. Photoreceptor cells are specialized neurons that sense light, and include two broad classes, rod cells and cone cells. Rod cells are involved in dark adaptation or vision in low light, while light adaptation or vision in bright light is caused by cone cells. Many neurodegenerative diseases that cause blindness (e.g., AMD) affect photoreceptor cells.

Extending from their cell bodies, photoreceptor cells morphologically have two distinct regions, the inner and outer segments. The outer segment, at the most distal end of the photoreceptor cell body, contains a disc-like structure that converts incident light energy into electrical impulses (photoconduction). The outer segment is attached to the inner segment by very fine and delicate cilia. The size and shape of the outer segment varies in rods and cones and is determined by the position in the retina. See Hogan, "Retina" inhistologyoftheamanEye: and atlas dTextBook (Hoganetal. (eds.). WBSaunders; Philadelphia, PA (1971)); EyeandOrbit, 8 ^thEd.，Bronetal.，(ChapmanandHall，1997)。

Ganglion cells are output neurons that transfer information from retinal interneurons (including horizontal cells, bipolar cells, amacrine cells) to the brain. Bipolar cells are named according to their morphology and accept input from photoreceptor cells, connect with amacrine cells, and send output radially to ganglion cells. Amacrine cells have processes parallel to the plane of the retina and typically inhibit export to ganglion cells. Amacrine cells are often subdivided into neurotransmitters or neuromodulators or peptides (e.g., calretin and calbindin) and interact with each other, bipolar cells, and photoreceptors. Bipolar cells are retinal interneurons, named morphologically; bipolar cells receive input from the photoreceptor cells and send input to the ganglion cells. Horizontal cells modulate and transform visual information obtained from a large number of photoreceptor cells and integrate horizontally (bipolar cells transmit information radially through the retina).

Other retinal cells that may be present in the retinal culture cells described herein include glial cells, such as muller glia cells and retinal pigment epithelial cells (RPEs). Glial cells surround cell bodies and axons. Glial cells do not carry electrical impulses and contribute to the maintenance of normal brain function. Miller glia, the major type of glial cell within the retina, provides support for retinal structure and is associated with retinal metabolism (e.g., contributing to regulation of ion concentration, degradation of neurotransmitters, and the elimination of some metabolites (see, e.g., kljavinet al, j. neurosci.11:2985 (1991)). miller fibers (also known as retinal support fibers) are the supporting glial cells of the retina, which traverse the thickness of the retina, from the inner limiting membrane to the rod and cone bases, where they form a line of complex connectors.

Retinal Pigment Epithelial (RPE) cells constitute the outermost layer of the retina, which is separated from the blood-vessel-rich choroid by bruch's membrane (basement choroid). The retinal pigment epithelium is a type of phagocytic epithelium that has some macrophage-like function, directly beneath the retinal photoreceptor cells. The back of the retinal pigment epithelium is closely juxtaposed to the end of the rod cell, and the disc-like structure that is detached from the outer segment of the rod cell is digested and absorbed by the retinal pigment epithelium. A similar process occurs in the discoid structure of the visual cone. Retinal pigment epithelial cells also produce, store and transport a variety of elements that aid in the normal function and survival of photoreceptor cells. Another function of retinal pigment epithelium is to recover vitamin a because it moves between the photoreceptor cells and the retinal pigment epithelium in a process called photopic and scotopic adaptation of the visual cycle.

Described herein are exemplary in vitro long-term cell culture systems and promote survival of cultured mature retinal cells (including retinal neurons) in culture for at least 2-4 weeks, more than 2 months, or up to 6 months. Cell culture systems are useful for identifying and labeling alkynyl phenyl linked amine derivative compounds for use in the methods described herein for treating and/or preventing ophthalmic diseases or disorders, or preventing or inhibiting the accumulation of lipofuscin and/or A2E in the eye. Retinal cells are isolated from non-embryonic, non-tumorigenic tissues and are not subjected to any means (e.g., transformation or infection with oncogenic viruses) to allow their unlimited growth and differentiation. The cell culture system includes all major retinal nerve cell types (photoreceptor, bipolar, horizontal, amacrine, and ganglion cells) and may also include other mature retinal cells such as retinal pigment epithelium and muller glia.

For example, a blood sample can be obtained from a subject, and the different retinoid compounds and the level of one or more retinoid compounds in the sample can be separated and analyzed by normal phase High Pressure Liquid Chromatography (HPLC) (e.g., using HP1100HPLC and Beckman, ultra-sphere silica (Ultrasphere-Si), 4.6 mm x 250 mm column, flow rate 1.4 ml/min, 10% ethyl acetate/90% hexane). Retinoids can be detected, for example, at a wavelength of 325nm using a diode array detector and using Hewlett packard chemical workstation A.03.03 (HPChemstation. 03.03) software. Excess retinoid can be determined, for example, by comparison of the profile of the retinoid in a sample from a normal subject (i.e., qualitatively, as the identity of the particular compound, and quantitatively, as the level of each particular compound). Those skilled in the art familiar with these analyses and techniques will readily appreciate that appropriate controls are also included.

As used herein, an increased or excess amount of an endogenous retinoid, such as 11-cis-retinol or 11-cis-retinal, refers to a higher level of endogenous retinoid than the concentration of endogenous retinoid found in healthy eyes of young vertebrates of the same population. The application of the alkynylphenyl-linked amine derivative compound reduces or eliminates the need for endogenous retinoids.

4. In vivo and in vitro methods for determining the efficacy of a compound

In one embodiment, there is provided a method of enhancing or prolonging survival of retinal cells (including survival of retinal nerve cells and survival of retinal pigment epithelial cells) using a compound described herein. Also provided herein are methods of inhibiting or preventing retinal cell degeneration, including retinal nerve cells (e.g., photoreceptor cells, amacrine cells, horizontal cells, bipolar cells, and ganglion cells), and other mature retinal cells, such as retinal pigment epithelial cells and muller glia cells, using the compounds described herein. These methods include: in certain embodiments, an alkynyl phenyl-linked amine derivative compound described herein is administered. Such compounds are useful for enhancing the survival of retinal cells, including photoreceptor cell survival and retinal pigment epithelial cell survival, inhibiting or delaying retinal cell degeneration, thereby increasing retinal cell viability, which as described herein may result in delaying or halting the progression of an ophthalmic disease or disorder or retinal damage.

The effect of the alkynyl phenyl-linked amine derivative compounds on retinal cell survival (and/or retinal cell degeneration) can be determined by cell culture models, animal models, and other methods described herein and used by those of skill in the art. Non-limiting examples of such assays include those described in the following references: oglivie et al, exp. neurol.161:675-856 (2000); U.S. patent No. 6,406,840; WO 01/81551; WO 98/12303; U.S. patent application No. 2002/0009713; WO 00/40699; U.S. Pat. nos. 6,117,675; U.S. patent No. 5,736,516; WO 99/29279; WO 01/83714; WO 01/42784; U.S. patent No. 6,183,735; U.S. patent No. 6,090,624; WO 01/09327; U.S. patent No. 5,641,750; U.S. patent application publication No. 2004/0147019; and U.S. patent application publication No. 2005/0059148.

The compounds described herein that may be used to treat ophthalmic diseases or disorders, including retinal diseases or disorders, may inhibit, prevent, slow, or interfere to some extent with one or more steps in the visual cycle, also referred to herein and in the art as the retinoid cycle. Without wishing to be bound by any particular theory, the amine derivative to which the alkynylphenyl group is attached may inhibit or prevent the isomerization step in the visual cycle, for example, by inhibiting or preventing the functional activity of trans-cis isomerase in the visual cycle. The compounds described herein may inhibit the conversion of all-trans retinol to 11-cis retinol, either indirectly or directly. The compound may adhere to or interact in some manner with the isomerase enzyme and inhibit the isomerase enzyme in at least one retinal cell. Any of the compounds described herein may also directly or indirectly inhibit or reduce the activity of an isomerase enzyme involved in the visual cycle. The compounds may prevent or inhibit the adhesion of isomerase to one or more substrates, including but not limited to all-trans retinyl ester substrates or all-trans retinol. Alternatively, or in addition, the compound may adhere to a catalytic site or region of an isomerase, thereby inhibiting the catalytic ability of the enzyme to isomerise at least one substrate. Based on scientific data, it is believed that at least one isomerase enzyme that catalyzes the isomerization of a substrate is located in the cytoplasm of RPE cells during the visual cycle. As described herein, the various steps, enzymes, substrates, intermediates, and products of the visual cycle have not been elucidated. However, a polypeptide known as RPE65, which has been found at the cytoplasmic-membrane boundary in RPE cells, is presumed to have isomerase activity (also known in the art as having isomerohydrolase activity) (see, e.g., Moiseyevetal, Proc. Natl. Acad. Sci. USA102:12413-18 (2004); Chenet al, Invest. Ophthalmol. Vis. Sci.47:1177-84(2006)), and other skilled in the art believe that RPE65 functions primarily as an all-trans-retinyl ester chaperone (see, e.g., Lambert. supra).

Exemplary methods will be described herein, and one skilled in the art will determine the level of enzymatic activity of an isomerase of the visual cycle in the presence of any of the compounds described herein. Compounds that reduce isomerase activity are useful for treating ophthalmic diseases or disorders. Thus, provided herein is a method for detecting inhibition of isomerase activity, the method comprising contacting a biological sample comprising an isomerase with an amine derivative compound linked to an alkynylphenyl group as described herein (i.e., mixing, combining or in some way allowing the compound and isomerase to interact), and determining the level of isomerase activity. One skilled in the art will appreciate that, by way of comparison, the level of activity of the isomerase in the absence of the compound or in the presence of a compound known not to alter the enzymatic activity of the isomerase can be determined and compared to the level of activity in the presence of the compound. A decrease in the level of isomerase activity in the presence of the compound compared to the level of isomerase activity in the absence of the compound indicates that the compound is useful in the treatment of an ophthalmic disease or disorder, such as age-related macular degeneration or macular disease. A decrease in the level of isomerase activity in the presence of the compound compared to the level of isomerase activity in the absence of the compound indicates that the compound is also useful in the methods described herein for inhibiting or preventing dark adaptation, inhibiting the formation of new blood vessels, and reducing hypoxia, for treating an ophthalmic disease or disorder, for example, diabetic retinopathy, diabetic maculopapulosis, retinal vessel occlusion, retinopathy of prematurity, or retinal ischemia reperfusion injury.

The ability of the alkynylphenyl-linked amine compounds described herein to inhibit or prevent dark adaptation of rod photoreceptors by inhibiting rhodopsin regeneration can be determined by in vitro testing and/or in vivo experiments in animal models. For example, inhibition of regeneration may be determined in a mouse model of a chemically induced diabetic condition or in a mouse model of diabetes (see, e.g., Phippsetal, invest, Ophthalmol. Vis. Sci.47:3187-94 (2006); Ramseyetal, invest, Ophthalmol. Vis. Sci.47:5116-24 (2006)). The level of rhodopsin in the animal retina (the first level) is measured (e.g., using spectrophotometric analysis) prior to administration of the agent and compared to the level of rhodopsin in the animal retina (the second level) measured after administration of the agent. A decrease in the second level of rhodopsin compared to the first level of rhodopsin indicates that the agent inhibits rhodopsin regeneration. One skilled in the art can readily determine and conduct appropriate comparisons and studies designed to determine whether rhodopsin regeneration is inhibited in a statistical or biological manner.

Methods and techniques for determining or characterizing the effect of any of the compounds described herein on dark adaptation and rod rhodopsin regeneration in mammals, including humans, can be accomplished according to the procedures described herein and practiced in the art. For example, the detection of visual stimulus versus time after illumination (i.e., photobleaching) in the dark can be determined before the administration of the first dose of the compound and for a period of time after the administration of the first dose and/or any subsequent doses. A second method of determining the prevention or inhibition of rod photoreceptor dark adaptation includes amplitude measurements of at least one, at least two, at least three, or more electroretinogram components, including, for example, a-waves and b-waves. See, e.g., Lamb et al, supra; asi et al, DocumentaOphthalmologica79:125-39 (1992).

Inhibition of rhodopsin regeneration by the alkynylphenyl linked amine compounds described herein includes reducing the level of chromophore (11-cis-retinal produced and present in PRE cells), and thus reducing the level of 11-cis retinal present in photoreceptor cells. Thus, when the compound is contacted with the retina under appropriate conditions and for a time sufficient to prevent rod photoreceptor dark adaptation and inhibit rhodopsin regeneration in rod photoreceptors, the compound causes a decrease (i.e., a statistically or biologically decrease) in the level of 11-cis-retinal in rod photoreceptors. That is, the level of 11-cis-retinal in the photoreceptor cell is higher before administration of the compound compared to the level of 11-cis-retinal in the photoreceptor cell after administration of the first dose of the compound and/or any subsequent dose. The effect of the compound can be monitored by measuring a first level of 11-cis retinal prior to administration of the compound and a second level of 11-cis retinal after administration of the compound at the first dose or any subsequent dose. A decrease in the second level compared to the first level indicates that the compound inhibits rhodopsin regeneration, thereby inhibiting or preventing dark adaptation of rod photoreceptors.

An example method of determining or characterizing the ability of an alkynyl phenyl linked amine compound to reduce retinal hypoxia includes testing retinal oxygenation, for example, by Magnetic Resonance Imaging (MRI), to measure changes in oxygen pressure (see, e.g., luanetal, invest. Methods are also within the art and are routinely practiced for determining or characterizing the ability of the compounds described herein to inhibit retinal cell degeneration.

Animal models can be used to characterize and identify compounds useful for treating retinal diseases and disorders. Ambati et al have described a recently developed animal model that can be used to evaluate treatments for macular degeneration (nat. med.9:1390-97 (2003); Epub2003 ℃ t 19). This animal model is one of the only few exemplary animal models currently available for evaluating compounds or any molecules for use in the treatment (including prevention) of the progression or development of a retinal disease or disorder. Animal models in which the ABCR gene encoding the ATP-binding cassette transporter (ATP-binding cassette transporter) is located at the edge of the discoid structure of the photoreceptor outer segment can be used to assess the response of compounds. Mutations in the ABCR gene are associated with Stargardt disease, while mutations in ABCR hybrids are associated with AMD. Thus, animals that have had some or all of their ABCR function lost can be used to characterize the alkynyl phenyl linked amine compounds described herein. (see, e.g., Mataeal, Invest, Ophthalmol, Sci.42:1685-90 (2001); Wengel, Cell98:13-23 (1999); Mataeal, Proc. Natl. Acad. Sci.USA97:7154-49 (2000); US 2003/0032078; U.S. Pat. No. 6,713,300). Other animal models include the use of mutant ELOVL4 transgenic mice to determine rhodopsin accumulation, electrophysiology and photoreceptor degeneration, or to prevent or inhibit accumulation or degeneration thereof (see, e.g., karanal, proc. natl. acad. sci. usa102:4164-69 (2005)).

The effect of any of the compounds described herein can be measured in an animal model of diabetic retinopathy, as described by Luan et al, or can be measured in a normal animal model in which the animal has been light-or dark-adapted in the presence and absence of any of the compounds described herein. Another exemplary method of determining the ability of an agent to reduce retinal hypoxia measures retinal hypoxia by deposition of a hydroxyl probe (see, e.g., deGooyeretal. (invest. ophthalmol. vis. sci.47:5553-60 (2006)). This technique may also be performed in animal models using Rho-/Rho-knockout mice (see deGooyer et al, supra) in which at least one compound described herein is administered to a group of animals in which at least one compound is present and absent, or may be performed in normal, wild-type animals in which at least one compound described herein is administered to a group of animals in which at least one compound is present and absent. Other animal models include models for measuring photoreceptor cell function, such as the rat model for measuring the oscillating potential of electronic retinal imaging (ERG) (see, e.g., Liu et al, invest. Ophtalmol. Vis. Sci.47:5447-52 (2006); Akula et al, invest. Ophtalmol. Vis. Sci.48:4351-59 (2007); Liu et al, invest. Ophtalmol. Vis. Sci.47:2639-47 (2006); Dembinka et al, invest. Ophtalmol. Vis. Sci.43:2481-90 (2002); Penn et al, invest. Ophtalmol. Vis. Sci.35:3429-35 (1994); Hancock et al, Ophtalmol. Vis. Sci.45:1002 (1008)).

As described herein and known in the art, methods for determining the effect of a compound on isomerase activity can be performed in vitro (Stecheret al, JBiolChem274:8577-85 (1999); see also Golczaketal, Proc. Natl. Acad. Sci. USA102:8162-67 (2005)). Retinal Pigment Epithelial (RPE) microsomal membranes isolated from animals (e.g., cattle, pigs, humans) can be used as the source of isomerase. The ability of amine derivative compounds having an alkynylphenyl group attached thereto to inhibit isomerase can also be determined by the murine in vivo isomerase assay. It is known that brief exposure of the eye to intense light (either "photobleaching" or simply "bleaching" of the visual pigment) can photoisomerize nearly all of the 11-cis-retinal in the retina. The recovery of 11-cis-retinal after bleaching can be used to assess isomerase activity in vivo (see, e.g., Maedaetal., J.neurochem85:944-956(2003); VanHooseltal., JBiolChem277:19173-82, 2002). Electroretinographic (ERG) recording (electroretinographic (ERG) recording) as previously described (Haeselesereal, Nat. Neurosci.7:1079-87(2004); Sugitomoet, J. Toxicol.Sci.22suppl2:315-25(1997); Keratital, Documenta Ophthalmologica100:77-92(2000)) may be performed. See also Deignetetal, Science,244: 968-.

Cell culture methods are also useful for determining the effect of the compounds described herein on retinal neuronal cell survival. Exemplary cell culture models are described herein and in detail in U.S. patent application publication No. US2005-0059148 and U.S. patent application publication No. US2004-0147019 (the entire contents of which are incorporated by reference) for determining the ability of an alkynyl phenyl linked amine-derived compound described herein to enhance or prolong the survival of nerve cells, particularly retinal nerve cells and retinal pigment epithelial cells, and to inhibit, prevent, slow or block degeneration of the eye or retina or retinal cells or retinal pigment epithelial cells thereof, and to determine which compounds are beneficial for treating ophthalmic diseases and disorders.

Cell culture models include long-term or expanded cultures of mature retinal cells, including retinal nerve cells (e.g., photoreceptor cells, amacrine cells, ganglion cells, horizontal cells, and bipolar cells). Cell culture systems and methods of producing cell culture systems provide for the expanded culture of photoreceptor cells. The cell culture system may also include Retinal Pigment Epithelium (RPE) cells and muller glia cells.

The retinal cell culture system may also include a cell stressor. The presence or use of cellular stressors infects mature retinal cells (including retinal nerve cells) in vitro in a manner that is beneficial for the pathological study of the observed retinal disease or disorder. The cell culture model provides an in vitro neural cell culture system that will facilitate the identification and biological testing of alkynyl phenyl linked amine derivative compounds that are useful in the treatment of neurological diseases or disorders in general, and degenerative diseases of the eye and brain in particular. The ability to maintain initial survival over a prolonged period of time in cultured cells in vitro derived from mature retinal tissue including retinal neurons in the presence of stressors enables testing of cell-to-cell interactions, selection and analysis of compounds and materials that stimulate nerves, use of control cell culture systems for in vitro Central Nervous System (CNS) and ophthalmic assays, and analysis of the effects on individual cells derived from a consistent retinal cell population.

Cell culture systems and retinal cell stress models including cultured mature retinal cells, retinal neurons and retinal cell stressors are useful for screening and characterizing alkynyl phenyl-linked amine derivative compounds capable of inducing or stimulating regeneration of Central Nervous System (CNS) tissues that have been damaged by disease. The cell culture system provides a mature retinal cell culture medium that is a mixture of mature retinal neural cells and non-retinal neural cells. The cell culture system includes all major retinal nerve cell types (photoreceptor, bipolar, horizontal, amacrine, and ganglion cells) and may also include other mature retinal cells such as retinal pigment epithelium and muller glia. By implanting these different types of cells into an in vitro cell culture system, the system essentially resembles an "artificial organ", which more closely resembles the natural state of the retina in vivo.

Viability of one or more mature retinal cell types isolated (obtained) from retinal tissue and tissue culture can be maintained for extended periods of time, for example, from 2 weeks to 6 months. The viability of retinal cells may be determined according to methods described herein and well known in the art. Like conventional nerve cells, retinal nerve cells do not undergo active cell division in vivo, and thus cell division of retinal nerve cells is not necessarily an indicator of viability. The advantage of the cell culture system is the ability to culture amacrine cells, photoreceptor cells and associated ganglion projection neurons and other mature retinal cells for extended periods of time, thereby providing an opportunity to determine the effectiveness of the alkynyl phenyl linked amine derivative compounds described herein for the treatment of retinal diseases.

The biological source of retinal cells or retinal tissue can be a mammal (e.g., a human, non-human primate, ungulate, rodent, canine, porcine, bovine, or other type of mammal), avian or other genus. In addition to adult or post-partum retinal tissue, retinal cells including retinal neurons derived from post-partum non-human primates, post-partum pigs, or post-partum chickens may be used for this retinal cell culture system.

In some cases, the cell culture system may provide for relatively long-term retinal cell survival, without including cells obtained or isolated or purified from non-retinal tissue. Such cell culture systems include cells isolated from the retina of the eye alone so as to be substantially free of various types of cells originating from other parts or regions of the eye separate from the retina, such as the ciliary body, iris, choroid, and vitreous. Other cell culture methods include the addition of non-retinal cells, such as ciliary body cells and/or stem cells (which may or may not be retinal stem cells) and/or other purified glial cells.

The in vitro retinal cell culture system described herein can serve as a physiological retinal model that can be used to characterize physiological aspects of the retina. This physiological retinal model can also be used as a more general model of neurobiology. The cell stressor may be included in a model cell culture system. The cell stressors described herein are retinal cell stressors that adversely affect viability or reduce viability of one or more different types of retinal cells (including retinal nerve cell types) in the cell culture system. Those skilled in the art will readily understand and appreciate the comparison of retinal cells cultured in an appropriate control cell system (e.g., a cell culture system without a cell stressor as described herein). Retinal cells that exhibit reduced viability as described herein means that the retinal cells have a reduced or decreased time to survive in a cell culture system (decreased lifespan) and/or exhibit a decrease, inhibition, or adverse effect on biological or biochemical functions (e.g., reduced or abnormal metabolism; onset of apoptosis; etc.). The reduction in retinal cell viability may be indicated by the following phenomena: cell death; a change or alteration in cellular structure or morphology; induction and/or progression of apoptosis; initiation, enhancement and/or acceleration of retinal neuronal neurodegenerative diseases (or neuronal damage).

Methods and techniques for determining cell viability are described in detail herein and are familiar to those skilled in the art. These methods and techniques for determining cell viability are useful for monitoring the health and status of retinal cells in cell culture systems, and for determining the ability of an alkynyl phenyl linked amine derivative compound described herein to alter (preferably increase, prolong, potentiate, improve) retinal cell or retinal pigment epithelial cell viability or retinal cell survival.

The addition of a cell stressor to a cell culture system can be used to determine the ability of an alkynylphenyl-linked amine derivative compound to abrogate, inhibit, eliminate, or mitigate the effects of the stressor. Retinal cell culture systems can include a cell stressor that is chemical (e.g., A2E, cigarette smoke concentrate); biological (e.g., toxin exposure; beta-amyloid; lipopolysaccharide) or non-chemical, such as physical, environmental or mechanical stressors (e.g., increasing pressure or exposure to light) (see, e.g., US 2005-0059148).

The retinal cell stressor model system may also include cell stressors, such as, but not limited to, stressors that may be risk factors for the disease or disorder or stressors that may contribute to the development or progression of the disease or disorder, including, but not limited to, light of different wavelengths and intensities; A2E; exposure to cigarette smoke condensate; oxidative stress (e.g., stress associated with the presence or contact of hydrogen peroxide, nitroprusside, Zn + + or Fe + +); increasing pressure (e.g., atmospheric or hydrostatic), glutamic acid or a glutamic acid agonist (e.g., N-methyl-D-aspartic acid (NMDA), alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA); kainic acid; quisqualic acid; amanitic acid; quinolinic acid, aspartic acid ester; trans-1-aminocyclopentyl-1, 3-dicarboxylate (ACPD)); amino acids (e.g., aspartic acid, L-cysteine; β -N-methylamino-L-alanine); heavy metals (e.g., lead); various toxins (e.g., mitochondrial toxins (e.g., malonate, 3-nitropropionic acid; rotenone, cyanide); MPTP (1-methyl-4-phenyl-1, 2,3,6, -tetrahydropyridine), which is metabolized to the active toxic metabolite MPP + (1-methyl-4-phenylpyridine)); 6-hydroxydopamine; alpha-synuclein; protein kinase C activators (e.g., phorbol myristate acetate); biological amino stimulants (e.g., methamphetamine, MDMA (3-4 methylenedioxymethamphetamine)); or a combination of one or more stressors. Retinal cell stressors that may be used include those that mimic neurodegenerative diseases affecting any one or more of the mature retinal cells described herein. Chronic disease models are particularly important because most neurodegenerative diseases are chronic. By using this in vitro cell culture system, the earliest events in the development of long-term disease may be detected by long-term cell analysis.

The retinal cell stressor may alter (i.e., increase or decrease in a statistical sense) the viability of retinal cells, such as by altering the viability of retinal cells (including retinal nerve cells and retinal pigment epithelial cells), or by altering the neurodegeneration of retinal nerve cells and/or retinal pigment epithelial cells. Preferably, the retinal cell stressor adversely affects retinal nerve cells or retinal pigment epithelial cells such that retinal nerve cell or retinal pigment epithelial cell viability is reduced or adversely affected (i.e., the time for cell survival in the presence of the stressor is reduced) or neurodegeneration of the cells (or nerve cell damage) is increased or enhanced. Stressors may affect only a single retinal cell type in the retinal cell culture medium, or stressors may affect two, three, four, or more different cell types. For example, stressors can alter the viability and viability of photoreceptor cells without affecting all other major cell types (e.g., ganglion cells, amacrine cells, horizontal cells, bipolar cells, retinal pigment epithelium, and muller glia). Stressors may shorten the survival time of retinal cells (in vivo or in vitro), increase the rate or extent of neurodegeneration of retinal cells, or in some other way adversely affect the viability, morphology, maturation or longevity of retinal cells.

The effect of a cell stressor on retinal cell viability in a cell culture system (in the presence and absence of an alkynylphenyl-linked amine derivative compound) can be determined from one or more different retinal cell types. The determination of cell viability may comprise assessing the structure and/or function of retinal cells at successive time intervals or at a particular time point after the retinal cell culture system is prepared. Viability or long-term survival of one or more different types of retinal cells or one or more different types of retinal neural cells may be measured before a morphological or structural change is observed based on one or more biochemical or biological parameters indicative of reduced viability, e.g., apoptosis or reduced metabolic function.

Chemical, biological, or physical cell stressors can reduce the viability of one or more types of retinal cells present in the cell culture system when added to the cell culture medium under the conditions described herein to maintain long-term cell culture. Alternatively, one or more culture conditions may be adjusted so that the effect of the stressor on the retinal cells can be more easily observed. For example, the concentration or percentage of fetal bovine serum in the cell culture medium may be reduced or eliminated when the cells are exposed to a particular cellular stressor (see, e.g., US 2005-0059148). Alternatively, retinal cells cultured in media containing specific concentrations of serum to maintain the cells may be suddenly exposed to media that does not contain any serum levels.

The retinal cell culture medium may be continuously exposed to the cell stressor for a period of time that depends on a reduction in viability of one or more retinal cell types in the retinal cell culture system. These cells are exposed to cellular stressors that immediately contact the surface of the retinal cells after detachment from the retinal tissue. Alternatively, the retinal cell culture may be exposed to the stressor after the culture is established or at any time thereafter. When two or more cell stressors are included in the retinal cell culture system, each stressor may be added to the retinal cell culture system at the same time and for the same length of time, or separately at different time points and for the same length of time or for different lengths of time. The amine compound having an alkynylphenyl group attached thereto can be added before the retinal cell culture is exposed to the cellular stressor, can be added simultaneously with the cellular stressor, or can be added after the retinal cell culture is exposed to the stressor.

Photoreceptor cells can be identified by using antibodies that specifically bind to photoreceptor cell-specific proteins, such as opsin, peripherin, and the like. By using discoid neuronal markers, the photoreceptor cells in the cell culture medium can also be determined by immunocytochemistry tracing the morphology of a subset of cells, or by morphology enhancing the image contrast of a viable medium. The outer segment can be found morphologically as part of the photoreceptor cell.

Retinal cells, including photoreceptor cells, can also be detected by functional analysis. For example, electrophysiological methods and techniques can be used to test the response of photoreceptor cells to light. Photoreceptor cells exhibit particular kinetics in a graded response to light. Calcium sensitive dyes can also be used to detect the grading response of media containing active photoreceptor cells to light. To analyze stress-inducing compounds or potential neural therapies, retinal cell culture media can be processed for immunocytochemistry, and photoreceptor cells and/or other retinal cells can be counted manually or by computer software using photomicrographic and imaging techniques. Other immunoassays known in the art (e.g., ELISA, in vitro) can also be used to identify and characterize retinal cells and retinal nerve cells of the cell culture model systems described herein.

Retinal cell culture stress models can also be used for direct and indirect recognition of pharmacological agents that act upon the bioactive agent of interest, such as the alkynyl phenyl-linked amine derivative compounds described herein. For example, the addition of a bioactive agent to a cell culture system in the presence of one or more retinal cell stressors can stimulate one cell type in a manner that enhances or reduces the viability of other cell types. Cell/cell interactions and cell/extracellular component interactions may be important in understanding disease and drug action mechanisms. For example, one nerve cell type may secrete trophic factors that affect the growth or survival of other types of nerve cells (see, e.g., WO 99/29279).

In another embodiment, the alkynyl phenyl linked amine-derived compound is implanted into a screening analyte comprising a retinal cell culture stress model system as described herein to determine whether and/or at what level or extent the compound increases the viability of a plurality of retinal cells (i.e., increases in a statistically or biologically significant manner). It will be readily understood and appreciated by those skilled in the art that the retinal cells described herein exhibit an increase in viability compared to retinal cells cultured in an appropriate control cell system (e.g., a cell culture system described herein in the absence of the compound), meaning that the retinal cells have an increased time to survive (increased lifespan) and/or maintain biological or biochemical function (normal metabolism and function of cellular organelles, lack of apoptosis, etc.) in the cell culture system. The increase in retinal cell viability may be caused by a delay in cell death or a reduction in the number of dead or dying cells; maintenance of structure and/or morphology; lack or delayed onset of apoptosis; a neurodegenerative disorder that delays, inhibits, slows progression of, and/or abrogates retinal nerve cells; or postponing or abrogating or preventing the effects of nerve cell damage. Methods and techniques for determining retinal cell viability, and thus whether retinal cells exhibit increased viability, are described in more detail herein and are known to those of skill in the art.

In certain embodiments, methods of determining whether an alkynyl phenyl linked amine derivative compound increases the survival of a photoreceptor cell are provided. One method includes contacting an alkynyl phenyl-linked amine compound described herein with a retinal cell culture system under conditions and for a time sufficient to allow retinal neural cells and the compound to interact. Measurement of enhanced survival (prolongation of survival) can be according to the methods described herein and known in the art, including detection of rhodopsin expression.

Amine derivative compounds having an alkynylphenyl group attached thereto increase retinal cell viability and/or enhance, promote, or prolong cell survival (i.e., prolong the time of survival of retinal cells including retinal nerve cells); and/or the ability to impair, inhibit or hinder degeneration caused directly or indirectly by stress as described herein, can be determined by any of a variety of methods known to those of skill in the art. For example, changes in cell morphology in the presence and absence of a compound can be determined by visual inspection, such as by light microscopy, confocal microscopy, or other microscopy methods known in the art. For example, cell viability may also be determined by viable and/or non-viable cell counts. Immunochemical or immunohistochemical techniques (e.g., fixed cell staining or flow cytometry) can be used to identify and evaluate cytoskeletal structures (e.g., by using antibodies specific for cytoskeletal proteins (e.g., glial fibrillary acidic protein, fibronectin, actin, vimentin, tubulin, etc.) or to evaluate the expression of cell markers as described herein. The effect of an alkynylphenyl-linked amine-derived compound on the integrity, morphology, and/or survival of cells can also be determined by measuring the phosphorylation state of a neuronal polypeptide, e.g., a cytoskeletal polypeptide (see, e.g., Sharmacetal, J.biol.chem.274:9600-06 (1999); liatal, J.Neurosci.20:6055-62 (2000)). Cell survival or cell death may also be determined according to methods described herein and known in the art for measuring apoptosis (e.g., annexin V binding, DNA fragmentation assays, caspase activation, marker assays, such as poly (ADP-ribose) polymerase (PARP), etc.).

In vertebrate eyes, e.g., mammalian eyes, the formation of A2E is a light-dependent process and its accumulation results in some adverse effects in the eye. This includes destabilization of the Retinal Pigment Epithelium (RPE) membrane, sensitization of cells to blue light damage and impaired deterioration of phospholipids. The product of A2E (and A2E related molecules) oxidized by molecular oxygen (ethylene oxide) was shown to induce DNA damage in cultured retinal pigment epithelial cells. All of these factors lead to a gradual decline in visual acuity and eventually blindness. If it were possible to reduce retinal formation during vision, this reduction would result in a reduction in the amount of A2E in the eye. Without being limited by theory, reducing the accumulation of A2E may reduce or delay the degenerative process of the retinal pigment epithelium and retina, thus slowing or preventing vision loss in dry AMD and stargardt disease.

In another embodiment, methods of treating and/or preventing degenerative diseases and disorders (including retinal neurodegenerative and ophthalmic diseases, and retinal diseases and disorders described herein) are provided. The subject in need of such treatment may be a human or non-human primate or other animal, and such subject may have developed symptoms of, or be at risk of developing, a retinal degenerative disease. The methods described herein provide for treating (including preventing or preventing) an ophthalmic disease or disorder by administering to a subject a composition comprising a pharmaceutically acceptable carrier and an alkynylphenyl-linked amine-derived compound (i.e., a compound having any one of the general formulae (a) - (G) and (I) - (III) and substructures thereof). As described herein, a method of increasing neural cells (e.g., retinal neural cells including photoreceptor cell survival) and/or inhibiting retinal neural cell degeneration by administering a pharmaceutical composition comprising an alkynyl phenyl linked amine derivative compound described herein is provided.

The enhanced survival (or prolonged or increased survival) of one or more retinal cell types in the presence of the alkynylphenyl-linked amine-derived compound indicates that the compound may be an effective agent for treating a degenerative disease, particularly a retinal disease or disorder, and including a retinal neurodegenerative disease or disorder. Cell viability and enhanced cell viability may be determined according to methods described herein and known to those skilled in the art, including viability assays and expression assays for detecting retinal cell marker proteins. To determine the enhanced survival of photoreceptor cells, opsin proteins, for example, including rhodopsin protein expressed by rods, may be detected.

In another embodiment, the subject is treated for stargardt disease or recessive (hereditary) macular degeneration. In stargardt disease associated with mutations in ABCA4 (also known as ABCR) transporters, accumulation of all-trans retinal has been implicated as the cause of lipofuscin pigment A2E formation, which is toxic to retinal cells and leads to retinal degeneration and hence blindness.

In another embodiment, the subject is treated for age-related macular degeneration (AMD). In various embodiments, the age-related macular degeneration may be of the dry or wet type. In age-related macular degeneration, blindness occurs primarily in the late stages of the complication when new blood vessels grow under the macula or the macula atrophies. Without being bound by any particular theory, the accumulation of all-trans retinal has been thought to be responsible for the formation of lipofuscin pigment N-retinylidene-N-retinylethanolamine (A2E) and A2E-related molecules, which are toxic to retinal pigment epithelium and retinal cells, and lead to retinal degeneration with eventual blindness.

A retinal neurodegenerative disease or disorder that is treated, prevented, ameliorated in symptoms, or slowed, inhibited, or stopped from developing by the compounds and methods described herein is a disease or disorder that causes or is characterized by loss of retinal nerve cells, which causes vision impairment. Such diseases or disorders include, but are not limited to, age-related macular degeneration (including both dry and wet forms of macular degeneration) and recessive (hereditary) macular dystrophy. The age-related macular degeneration described herein refers to a disease that affects the macular region (central region of the retina) and causes central vision to decline and blindness. Age-related macular degeneration commonly occurs in individuals over the age of 55. The etiology of age-related macular degeneration may include both environmental and genetic factors (see, e.g., Lyengaretal, am. J. hum. Genet.74:20-39 (2004)) (Epub2003December 19); Kenealyetal, mol. Vis.10:57-61 (2004); Gorinetal, mol. Vis.5:29 (1999)). Rarely, macular degeneration also occurs in young adults, including children and infants, and often these diseases are the result of genetic mutations. Types of juvenile macular degeneration include Stargardt's disease (see, e.g., Glazeretal, Ophthalmol. Clin. NorthAm.15:93-100, viii (2002); Wengel., Cell98:13-23 (1999)); dorne honeycomb retinal dystrophy (see, e.g., kermaniet, hum. genet.104:77-82 (1999)); sorsby fundus dystrophy, MalattiaLevinthese, yellow-spotted fundus and autosomal dominant hemorrhagic macular dystrophy (see also Seddonetal, Ophthalmology108:2060-67 (2001); Yatesetal, J.Med.Genet.37:83-7 (2000); Jaaksonetal, hum.Mutat.22:395-403 (2003)).

Geographic atrophy of the retinal pigment epithelium is an advanced form of non-neovascular dry-type age-related macular degeneration and is associated with atrophy of the choroid, retinal pigment epithelium, and retina.

Recessive (hereditary) macular degeneration belongs to recessive hereditary diseases, and is a hereditary blindness disease of children. The major pathological defect of stargardt disease is also the accumulation of toxic lipofuscin pigment, such as A2E, in Retinal Pigment Epithelial (RPE) cells. This accumulation appears to be responsible for photoreceptor cell death and severe vision loss found in individuals with stargardt disease. The compounds described herein may reduce the synthesis of 11-cis-retinal (11cRAL or retinal) and the regeneration of rhodopsin by inhibiting isomerase enzymes during the visual cycle. Photoactivation of rhodopsin results in the release of its all-trans retinal, which constitutes the first reactant in the biosynthesis of A2E. Treatment with an alkynyl phenyl linked amine derivative compound can inhibit lipofuscin accumulation, thereby delaying the onset of vision loss in individuals with Stargardt's disease and AMD, without toxicity that would preclude treatment with an alkynyl phenyl linked amine derivative compound. The compounds described herein may be useful in the effective treatment of other forms of retinal or macular degeneration associated with lipofuscin accumulation.

Administration of an alkynylphenyl-linked amine-derived compound to a subject can prevent the formation of lipofuscin pigment A2E (and A2E-related molecules), which is toxic to retinal cells and causes retinal degeneration. In some embodiments, administration of an alkynyl phenyl linked amine derivative compound can reduce production waste, such as lipofuscin pigment A2E (and A2E related molecules) improves the progression of AMD (e.g., dry form) and stargardt disease, and reduces or delays vision loss (e.g., choroidal neovascularization and/or chorioretinal atrophy). In previous studies, 13-cis-retinoic acid (Aiyoutong) was administered to subjectsOr Isotretinoin (Isotretinoin)), a drug commonly used for the treatment of acne and an inhibitor of 11-cis-retinol dehydrogenase, to prevent A2E accumulation in the retinal pigment epithelium. However, a major drawback of this proposed therapy is that 13-cis-retinoic acid can be readily isomerized to all-trans-retinoic acid. All-trans-retinoic acid is a powerful teratogenic compound that adversely affects the proliferation and development of cells. Vitamin a acid also accumulates in the liver and may be a cause of liver disease.

In other embodiments, the amine-derived compound having an alkynylphenyl group attached thereto is administered to a subject, such as a human having a mutation in the ABCA4 transporter in the eye. Amine derivative compounds having an alkynylphenyl group attached thereto can also be administered to elderly individuals. As described herein, an elderly human subject is typically at least 45 years of age, or at least 50 years of age, or at least 60 years of age, or at least 65 years of age. In stargardt disease, which is associated with mutations in the ABCA4 transporter, it has been suggested that accumulation of all-trans retinal is responsible for the formation of lipofuscin pigment A2E (and A2E-related molecules), which is toxic to retinal cells and leads to retinal degeneration and thus eventual blindness. Without being limited by theory, the alkynyl phenyl-linked amine derivative compounds described herein may be strong inhibitors of the isomerase enzymes involved in the visual cycle. Treatment of an individual with an amine derivative compound having an alkynylphenyl group attached as described herein can prevent or slow the formation of A2E (and A2E related molecules) and have protective properties for normal vision.

In other embodiments, one or more compounds described herein can be used to treat other ophthalmic diseases or disorders, for example, glaucoma, retinal detachment, hemorrhagic retinopathy, retinitis pigmentosa, inflammatory retinopathy, proliferative vitreoretinopathy, retinal dystrophy, hereditary optic neuropathy, Sorsby fundus dystrophy, ocular vitritis, retinal damage, optic neuropathy; and retinal disorders associated with other neurodegenerative diseases such as Alzheimer's disease, multiple sclerosis, Parkinson's disease or other neurodegenerative diseases affecting brain cells; retinal disorders associated with viral infections or other conditions, such as aids. Retinal disorders also include light damage to the retina, which is associated with increased light exposure (i.e., overexposure), such as, for example, accidental intense or intense light exposure during surgery; intense, violent, or prolonged exposure to sunlight, such as in desert or snowy terrain; in combat, for example, when a light bomb or explosion or laser device or the like is observed. The retinal disease may be of a degenerative or non-degenerative nature. Non-limiting examples of retinal degenerative diseases include age-related macular degeneration and recessive (hereditary) macular dystrophy. Examples of non-degenerative retinal diseases include, but are not limited to, hemorrhagic retinopathy, retinitis pigmentosa, optic neuropathy, inflammatory retinopathy, diabetic maculopapulosis, retinal vessel occlusion, retinopathy of prematurity, or retinal ischemia-reperfusion injury, proliferative vitreoretinopathy, retinal dystrophy, hereditary optic neuropathy, Sorsby's fundus dystrophy, uveitis, retinal damage, retinal disorders associated with alzheimer's disease, retinal disorders associated with multiple sclerosis, retinal disorders associated with parkinson's disease, retinal disorders associated with viral infection, retinal disorders associated with overexposure, and retinal disorders associated with aids.

In other embodiments, at least one compound described herein may be used to treat, prevent, ameliorate symptoms of, or slow, inhibit, or arrest the progression of certain ophthalmic diseases and disorders, including but not limited to diabetic retinopathy, diabetic maculopapulosis, diabetic macular edema, retinal ischemia reperfusion injury, and retinal vessel occlusion (including venous occlusion and arterial occlusion).

Diabetic retinopathy is a major cause of blindness in humans and is a complication of diabetes mellitus. Diabetic retinopathy occurs when diabetes damages blood vessels within the retina. Non-proliferative retinopathy is a common, usually mild form that generally does not impede vision. Abnormalities are limited to the retina, and vision is impaired only when spots are involved. If left untreated, retinopathy progresses to proliferative retinopathy, which is the more severe diabetic retinopathy. Proliferative retinopathy occurs when new blood vessels proliferate in and around the retina. As a result, bleeding into the vitreous, retinal edema, and/or retinal detachment may occur, resulting in blindness.

Other ophthalmic diseases and disorders that can be treated using the methods, compositions described herein include diseases, disorders, and conditions associated with, exacerbated by, or caused by intraretinal ischemia. Retinal ischemia includes both inner and outer retinal ischemia. Retinal ischemia can be caused by choroidal or retinal vascular diseases, such as central or branch retinal vision obstruction, collagen vascular diseases, and thrombocytopenic purpura. Retinal vasculitis and obstruction occur with retinal periphlebitis and systemic lupus erythematosus.

Retinal ischemia may be associated with retinal vessel occlusion. In the united states, branch and central retinal vein occlusions are the second most common retinal vascular disease after diabetic retinopathy. About 7% to 10% of individuals with retinal vein occlusion in one eye eventually become bilaterally infected. Visual field loss is usually caused by macular edema, ischemia, or vitreous hemorrhage following disk or retinal neovascularization induced by the release of vascular endothelial growth factor.

Arteriosclerotic changes in the retinal vein wall at the site of arteriovenous crossings of the retina (the region where the arteries and veins share a common adventitial cuff) result from the crossing arteries contracting. The contraction results in thrombosis and subsequent vein occlusion. Blocked veins can lead to macular edema and hemorrhage followed by a breakdown of the blood-retinal barrier in the area through which the veins flow, with blood interrupted by turbulent drainage venous blood flow disturbance circulation, endothelial cell damage and ischemia. Clinically, the appearance of feathery white plaques in ischemic areas of the retina is called cotton wool spots.

Ischemic branch retinal vein occlusion causes acute central and near central visual field loss corresponding to the location of the involved retinal quadrants. Retinal neovascularization due to ischemia can lead to vitreous hemorrhage and subacute or acute vision loss.

Two types of central retinal vein occlusion, ischemic and non-ischemic, can occur depending on whether there is widespread retinal ischemia. Even in the non-ischemic form, the macula may be ischemic. Approximately 25% of central retinal vein occlusions are ischemic. Diagnosis of central retinal vein occlusion can generally be made from characteristic ophthalmoscopy results, which include retinal hemorrhages, vein distensions, vein distortions, and cotton wool spots in all four quadrants. Macular edema and foveal ischemia can lead to vision loss. Extracellular fluid increases interstitial pressure, which may lead to areas of retinal capillary closure (i.e., lamellar ischemic retinal albinism) or ciliary retinal artery occlusion.

Individuals with ischemic central retinal vein occlusion are more likely to experience sudden onset of vision loss, and visual acuity below 20/200, versus widespread nonperfusion of afferent pupil defects, high intraretinal hemorrhages, and fluorescence angiography. The natural history of ischemic central retinal vein occlusion is associated with adverse consequences: eventually about two thirds of individuals with ischemic central retinal vein occlusion will have ocular neovascularization, while one third will have neovascularized glaucoma. The latter is a severe form of glaucoma that can lead to rapid visual field and vision loss, corneal epithelial edema with secondary epithelial erosion, and induction of bacterial keratitis, severe pain, nausea and vomiting, with eventual phthisis (atrophy without light sensation in the whole eye).

The subject (or objects) referred to herein may be any mammal, including a human, may be afflicted with a neurodegenerative disease or condition, including an ophthalmic disease or disorder, or may be detected disease-free. Thus, treatment may be administration to a subject with an existing disease, or treatment may be prophylaxis or administration to a subject at risk of developing a disease or condition. Treatment or therapy refers to any marker of success in treating or ameliorating an injury, condition or condition, including any objective or subjective parameter, such as reduction; reducing immunity; reduced symptoms or making the injury, condition or condition more tolerable to the individual; slowing the rate of degeneration or decline; the endpoint of degeneration is less debilitating; or improving the physical or mental well-being of the subject.

Treatment or amelioration of symptoms can be based on objective or subjective parameters, including the results of physical examination. Thus, the term "treatment" includes administration of a compound or agent described herein to treat pain, hyperalgesia, allodynia, or a painful event, and to prevent or delay, alleviate, or absorb or inhibit the development of symptoms or conditions associated with pain, hyperalgesia, allodynia, a painful event, or other disorders. The term "therapeutic effect" refers to the reduction, elimination, or prevention of a disease, a symptom of the disease, or a sequela of the disease in a subject. Treatment also includes measuring (e.g., testing over a period of weeks or months) the recovery or improvement of retinal neuronal function (including photoreceptor function) in the vertebrate visual system over a period of time, e.g., such as visual acuity and visual field tests, etc. Treatment also includes stabilizing the disease process (i.e., slowing, reducing or stopping the progression of ocular disease and associated symptoms) and minimizing other degeneration of the vertebrate visual system. Treatment also includes prophylaxis, meaning administration of an alkynyl phenyl linked amine-derivative compound to a subject to prevent degradation or further degradation or worsening or further worsening of the visual system of a vertebrate subject, and to prevent or inhibit the development and sequelae of disease and/or associated symptoms.

Various methods and techniques used by those skilled in the medical and ophthalmic arts to determine and assess disease status and/or monitor and evaluate treatment regimens include, for example, fluorescein angiography, fundus photography, indocyanine green dye tracking of the choroidal circulation system, ophthalmoscopy, Optical Coherence Tomography (OCT), and visual acuity testing.

Fluorescein angiography involves intravenous injection of a fluorescent dye and then observation of any dye leakage as it flows through the entire eye. Intravenous injection of indocyanine green dye may also be used to determine whether blood vessels in the eye are damaged, particularly in the choroidal circulatory system located just behind the retina. Fundus photography can be used to examine the optic nerve, macula, blood vessels, retina and vitreous. Microangiomas are visible lesions of diabetic retinopathy that can be found in digital images of the fundus at an early stage of the disease (see, e.g., U.S. patent application publication No. 2007/0002275). Funduscles may be used to examine the retina and vitreous. Fundus examination is typically performed with the pupil dilated to obtain an optimal field of view within the eye. Two types of ophthalmoscopes may be used: direct and indirect. Direct ophthalmoscopes are commonly used to view the optic nerve and the center of the retina. The peripheral or entire retina can be viewed by using an indirect ophthalmoscope. Optical Coherence Tomography (OCT) can yield high resolution, high speed, non-invasive cross-sectional images of body tissue. OCT is non-invasive and provides microscopic detection of early signs of tissue destruction.

By subject or subject is meant any vertebrate or mammalian subject or subject to which the compositions described herein are administered. The term "vertebrate" or "mammal" includes human and non-human primates, and laboratory animals such as rabbits, rats, chinchilla, and other animals, such as domestic pets (e.g., cats, dogs, horses), farm animals, and zoo animals. A subject in need of treatment with the methods described herein can be identified according to screening methods recognized in the medical arts for determining risk factors or symptoms associated with an ophthalmic disease or condition described herein, or for determining an existing ophthalmic disease state or condition in a subject. These and other conventional methods allow a clinician to select an individual in need of treatment using the methods and formulations described herein.

V. pharmaceutical composition

In certain embodiments, the amine derivative compound having an alkynylphenyl group attached thereto can be applied as a pure chemical. In other embodiments, the alkynylphenyl linked amine derivative compound can be combined with a pharmaceutical carrier (also referred to herein as a pharmaceutically suitable excipient)In combination with a vehicle (i.e., a pharmaceutically suitable and acceptable carrier, diluent, etc., which is a non-toxic, inert material that does not interact with the activity of the active ingredient)), the carrier/excipient is selected based on the chosen route of administration and standard pharmaceutical practice, e.g., as described in Remington: the science and practice of pharmacy (geno, 21) ^stEd. mackpub.co., Easton, PA (2005)), the entire disclosure of which is incorporated herein by reference.

Accordingly, provided herein is a pharmaceutical composition of a compound described herein, comprising one or more amine derivative compounds having an alkynylphenyl group attached thereto, or a crystalline form of a stereoisomer, a tautomer, a prodrug, a pharmaceutically acceptable salt, a hydrate, a solvate, an acid salt hydrate, a nitroxide, or an isomer thereof, and one or more pharmaceutically acceptable carriers, and optionally other therapeutic and/or prophylactic ingredients. A carrier is acceptable or suitable if it is compatible with the other ingredients of the composition and not deleterious to the recipient (i.e., the individual) of the composition. Pharmaceutically acceptable or suitable compositions include ophthalmically suitable or acceptable compositions.

Accordingly, another embodiment provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound having the structure of formula (a):

m is 0, 1, 2 or 3;

Y is a bond, -C (R)²⁷)(R²⁸) -or-C (R)²⁷)(R²⁸)-C(R²⁹)(R³⁰)-；

R³⁰and R³⁵Each independently is hydrogen or C₁-C₅An alkyl group.

Various embodiments further provide pharmaceutical compositions comprising a pharmaceutically acceptable excipient and a compound of any one of formulas (B) - (G) and (I) - (III):

wherein the structure is as defined in the foregoing and herein.

The pharmaceutical compositions (e.g., oral or injectable, or combined methods, or applied as eye drops) may be in liquid or solid form. The liquid pharmaceutical composition may include, for example, one or more of the following: sterile diluents such as water for injection, saline solutions, preferably physiological saline, ringer's solution, isotonic sodium chloride, fixed oils as solvent or suspending medium, polyethylene glycols, glycerol, propylene glycol or other solvents; an antibacterial agent; an antioxidant; a chelating agent; buffers and tonicity adjusting agents such as sodium chloride or dextrose. The gastrointestinal tract preparation can be sealed in ampoule, disposable syringe or multi-dose bottle made of glass or plastic. Physiological saline is commonly used as an excipient, and injectable pharmaceutical compositions or compositions for ocular delivery are preferably sterile.

The amine derivative compound having an alkynylphenyl group attached thereto can be administered to a human or other non-human vertebrate. In certain embodiments, the compound is substantially pure in that it contains less than about 5% or less than about 1%, or less than about 0.1% of other small organic molecules, e.g., intermediate contaminants or by-products as produced in one or more steps of the synthetic method. In other embodiments, it may be administered a combination of one or more amine derivative compounds having an alkynylphenyl group attached.

The alkynylphenyl-linked amine derivative compound can be delivered to the subject by any suitable means, including, for example, oral, parenteral, intraocular, intravenous, intraperitoneal, intranasal (or other methods of delivery to the mucosa, e.g., nasal, throat, and bronchial), or ocular topical administration, or by intraocular or periocular methods. Topical administration may include, for example, eye drops, intraocular injection, or periocular injection. Periocular injection typically involves the injection of an isomerization synthesis inhibitor, i.e., subconjunctival injection or injection of an alkynylphenyl-linked amine-derivative compound into the Tennon space (beneath the fibrous tissue covering the eye). Intraocular injection typically involves injecting an amine derivative compound having an alkynylphenyl group attached to the vitreous. In some embodiments, administration is non-invasive, such as by eye drop or oral dosage form, or by a combination method.

The amine derivative compound having the alkynylphenyl group attached thereto can be administered and formulated using a pharmaceutically acceptable (suitable) carrier or vehicle and techniques conventional in the art. Pharmaceutically acceptable or suitable carriers include ophthalmically suitable or acceptable carriers. The carrier is selected based on the solubility of the alkynylphenyl-linked amine derivative compound. Compositions suitable for ophthalmic use include those that can be topically applied to the eye, such as by eye drops, injection, and the like. In the case of an eye drop, the formulation may optionally include, for example, an ophthalmically compatible agent, such as an isotonic agent, for example, sodium chloride, concentrated glycerin, and the like; buffers such as sodium phosphate, sodium acetate, and the like; surfactants such as polyethylene glycol sorbitol monooleate (also known as Polysorbate80), polyoxyl stearate 40, polyoxyethylene hydrogenated castor oil, and the like; stabilizers, such as sodium citrate, sodium edetate, and the like; preservatives, such as benzalkonium chloride, parabens, and the like; and other ingredients. Preservatives may be employed, for example, at a level of from about 0.001 to about 1.0% weight/volume. The pH of the formulation is generally within a range acceptable for ophthalmic formulations, such as a pH of about 4 to 8, or a pH of about 5 to 7, or a pH of about 6 to 7, or a pH of about 4 to 7, or a pH of about 5 to 8, or a pH of about 6 to 8, or a pH of about 4 to 6, or a pH of about 5 to 6, or a pH of about 7 to 8.

For injection, the alkynylphenyl-linked amine derivative compound may be provided in the form of: in injection-grade saline solutions, in the form of injection-liposome solutions, sustained release polymer systems, and the like. Intraocular and periocular injections are known to those skilled in the art and are described in a number of publications, including, for example, Spaeth, ed., OphthalmicSurgery: principles of practice, w.b. sanders co., philiadelphia, Pa.,85-87,1990.

For compositions comprising at least one compound described herein for delivery by the mucosal route, including delivery to the nasal cavity, throat and trachea, the compositions may be delivered as an aerosol. The compounds may be delivered mucosally in liquid or powder form. For example, the composition may be delivered by a pressurized aerosol container with a suitable propellant, such as a hydrocarbon propellant (e.g., propane, butane, isobutene). The composition can be delivered by a non-pressurized delivery system, such as a nebulizer or atomizer.

Suitable oral dosage forms include, for example, tablets, pills, sachets, or hard or soft capsules, methylcellulose or other suitable material that readily disintegrates in the digestive tract. Suitable non-toxic solid carriers can be used and include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose Glucose, sucrose, magnesium carbonate, and the like. (see, e.g., Remington: the science and practice of pharmacy (Gennaro, 21)^stEd.MackPub.Co.,Easton，PA(2005))。

The alkynyl phenyl linked amine derivative compounds described herein may be used in sustained or slow release formulations. Such compositions can generally be prepared and administered using well-known techniques, for example, oral, periocular, intraocular, rectal, or subcutaneous implantation, or implantation at the intended target site. Sustained release formulations may include an agent dispersed in a carrier matrix and/or contained in a reservoir surrounded by a rate controlling membrane. The excipients used in such formulations are biocompatible and may also be biodegradable; preferred formulations provide a relatively constant level of release of the active ingredient. The amount of active compound in the sustained release formulation will depend on the site of implantation, the rate and expected time of release, and the nature of the condition being treated or prevented.

Systemic drug absorption of drugs or compositions administered by the ocular route is known to those skilled in the art (see, e.g., leeet al, int.j. pharm.233:1-18 (2002)). In one embodiment, the amine derivative compound having an alkynylphenyl group attached is delivered by topical ocular delivery methods (see, e.g., curr. drug metal.4: 213-22 (2003)). The composition may be in the form of an eye drop, ointment or salve, such as an aqueous eye drop, aqueous ophthalmic suspension, non-aqueous ophthalmic solution and non-aqueous ophthalmic suspension, gel, eye ointment, and the like. For preparing the gel, for example, carbopol, methyl cellulose, sodium alginate, hydroxypropyl cellulose, ethylene maleic anhydride polymer, and the like can be used.

The dosage of the composition comprising at least one alkynylphenyl linked amine derivative compound described herein can vary depending on the condition of the individual (e.g., human), i.e., the stage of disease, general health, age, and other factors used by those skilled in the medical arts to determine dosage. When the composition is used as an eye drop, for example, one to several drops, preferably 1 or 2 drops (about 50 μ l per drop), per unit dose, it may be used from about 1 to about 6 times per day.

The pharmaceutical compositions can be administered in a manner suitable for the treatment (or prevention) of the disease as determined by one of skill in the medical arts. The appropriate dosage and appropriate time and frequency of administration will depend on the following factors: the condition of the individual, the kind and severity of the individual's disease, the particular form of the active ingredient and the method of administration. In general, an appropriate dosage and treatment regimen provides a sufficient amount of the composition to facilitate treatment and/or prevention (e.g., to improve clinical outcome, such as more rapid total or partial remission, or longer periods of disease-free and/or overall survival, or reduction in severity of symptoms). For prophylactic use, the dose should be sufficient to prevent, delay the onset of, or reduce the severity of a disease associated with neurodegeneration of retinal nerve cells and/or degeneration of other mature retinal cells (e.g., RPE cells). The optimal dosage can generally be determined using experimental models and/or clinical trials. The optimal dosage depends on the body mass, body weight or blood volume of the individual.

The dose of the alkynyl phenyl-linked amine derivative compound may be appropriately selected depending on the clinical state, condition and age, the formulation, and the like of the subject. In the case of an eye drop, the alkynyl phenyl linked amine derivative compound can be administered, for example, in a single dose from about 0.01mg, about 0.1mg, or about 1mg to about 25mg, to about 50mg, to about 90 mg. The eye drops may be administered one or more times per day, as desired. In the case of injection, a suitable dose may be, for example, about 0.0001mg, about 0.001mg, about 0.01mg, or about 0.1mg to about 10mg, to about 25mg, to about 50mg, or to about 90mg of the alkynylphenyl-linked amine derivative compound once to seven times per week. In other embodiments, about 1.0 to about 30mg of the amine derivative compound having an alkynylphenyl group attached can be administered from once to seven times per week.

Oral dosages may typically range from 1.0 to 1000mg, once to four times daily, or more. Exemplary oral dosages range from 10 to 250mg, one to three times a day. If the composition is a liquid formulation, the composition contains at least 0.1% of the active compound (e.g., from 1.0 to 1000mg), for example, from about 2% to about 60%, by particular mass or weight per unit volume of carrier.

In certain embodiments, at least one of the alkynyl phenyl-linked amine compounds described herein can be administered under conditions and at one time, which can inhibit or prevent dark adaptation of rod photoreceptor cells. In some embodiments, such compounds are administered at least 30 minutes (half an hour), 60 minutes (one hour), 90 minutes (1.5 hours), or 120 minutes (2 hours) before the subject sleeps. In some embodiments, such a compound may be administered to a subject at night before the subject sleeps. In other embodiments, light stimuli may be blocked or removed during the day or under normal lighting conditions by placing the subject in an environment where light is removed, such as placing the subject in a dark room or by applying an eye mask over the subject's eyes. When the light stimulus is eliminated in this manner or other means contemplated in the art, the agent may be administered before sleep.

The dose of the compound to be administered to prevent or inhibit rod photoreceptor cell dark adaptation may be appropriately selected depending on the clinical state, condition and age of the subject, the dosage form, and the like. In the case of an eye drop, the compound (or composition comprising the compound) may be administered, for example, in a single dose of from about 0.01mg, about 0.1mg, or about 1mg to about 25mg, to about 50mg, to about 90 mg. In the case of injection, a suitable dose may be, for example, about 0.0001mg, about 0.001mg, about 0.01mg, or about 0.1mg to about 10mg, to about 25mg, to about 50mg, to about 90mg of the compound administered on any number of days between one and seven days per week before going to bed or before removing all light sources from the subject. In other embodiments, the compound is administered in a dose of between 1 and 10mg of compound per kilogram of body weight of the subject (i.e., for example, 80-800mg per dose for an 80kg subject) for administration via eye drop or injection. In other embodiments, about 1.0 to about 30mg of the compound may be administered from once to seven times per week. Oral dosages may typically range from about 1.0 to about 1000mg, administered anywhere between Mondays and seven days per week. An exemplary oral dosage is from about 10 to about 800mg once daily before bedtime. In other embodiments, the composition may be delivered by intravitreal administration.

Methods of making the compounds and pharmaceutical compositions described herein are also provided. Compositions comprising a pharmaceutically acceptable excipient or carrier and at least one alkynylphenyl linked amine derivative compound described herein can be synthesized according to any of the methods described herein or practiced in the art to yield the compound, and then formulated with the pharmaceutically acceptable carrier. The formulation of the composition will be appropriate depending on several factors, including but not limited to, the route of delivery, the dosage, and the stability of the compound.

Other embodiments and uses will be apparent to those skilled in the art in view of the present disclosure. The following examples are provided only to illustrate various embodiments and are not to be construed as limiting the invention in any way.

While preferred embodiments of the present invention are shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that numerous alternative variations to the embodiments described herein may be utilized in practicing the invention. It is intended that the following claims define the scope of the invention and that the method and structure within the scope of these claims and their equivalents be covered thereby.

Examples

Unless otherwise indicated, reagents and solvents used were obtained from commercial suppliers. Anhydrous solvents and baked glassware were used for synthetic transformations that are generally considered to be sensitive to moisture and/or oxygen. Flash column chromatography and Thin Layer Chromatography (TLC) were performed on silica gel unless otherwise indicated. As described above, proton and carbon nuclear magnetic resonance spectra were obtained in the following instruments: VarianVnmrS400 (proton spectra obtained at 400MHz and carbon spectra obtained at 100 MHz), Bruker AMX500 or 300 spectrometer (proton spectra obtained at 500 or 300MHz and carbon spectra obtained at 125 or 75 MHz). The spectra are given in ppm () and the coupling constant J is given in Hz. For proton spectra, either tetramethylsilane was used as internal standard or the solvent peak was used as reference peak. For the carbon spectra, the solvent peak was used as a reference peak. Chiral HPLC analysis was obtained using a chiralpak ia column (4.6mmx250mm,5 μ) and a diode array detector (dioderaraydetection). The flow rate was 1 mL/min.

Analytical HPLC method

Method 001A

Column: YMCODA-A (150 mm. times.4.6 mm. times.5 μ)

Flow rate: 1.2mL/min

Injection volume: 10 μ L

Column heater temperature (ColumnOventemp): 30 deg.C

Cell temperature: 40 deg.C

Wavelength: binary 220nm &254nm

Bandwidth: 4nm

Mobile phase:

A: water containing 0.05% TFA

B: acetonitrile containing 0.05% TFA

Operating time: for 10 min.

Gradient elution procedure

Time (min)	Flow rate of flow	%A	%B
				0.0	1.2	90	10
5.0	1.2	20	80
				7.0	1.2	20	80
7.01	1.2	90	10
				10.0	1.2	90	10

Diluent agent: acetonitrile/Water (1:1) with 0.05% TFA

Method 002A

Column: YMCODA-A (150 mm. times.4.6 mm. times.5 μ)

Flow rate: 1.2mL/min

Injection volume: 10 μ L

Column heater temperature: 30 deg.C

Cell temperature: 40 deg.C

Wavelength: binary 220nm &254nm

Bandwidth: 4nm

Mobile phase:

a: water containing 0.05% TFA

B: acetonitrile containing 0.05% TFA

Operating time: for 10 min.

Gradient elution procedure

Time (min)	Flow rate of flow	%A	%B
				0.0	1.2	100	0
5.0	1.2	50	50

Time (min)	Flow rate of flow	%A	%B
				7.0	1.2	50	50
7.01	1.2	100	0
				10.0	1.2	100	0

Diluent agent: acetonitrile/Water (1:1) with 0.05% TFA

Method 003A

Column: YMCODA-A (150 mm. times.4.6 mm. times.5 μ)

Flow rate: 1.2mL/min

Injection volume: 10 μ L

Column heater temperature: 30 deg.C

Cell temperature: 40 deg.C

Wavelength: binary 220nm &254nm

Bandwidth: 4nm

Mobile phase:

a: water containing 0.05% TFA

B: acetonitrile containing 0.05% TFA.

Operating time: for 10 min.

Gradient elution procedure

Time (min)	Flow rate of flow	%A	%B
				0.0	1.2	50	50
5.0	1.2	0	100
				7.0	1.2	0	100
7.01	1.2	50	50
				10.0	1.2	50	50

Diluent agent: acetonitrile/Water (1:1) with 0.05% TFA

Preparation method

Method 001P

Column: YMCODA-A (500 mm. times.30 mm. times.10 μ)

Flow rate: 30mL/min

Injection volume: 5mL

Column heater temperature: ambient temperature

Wavelength: binary 220nm

Mobile phase:

A: water containing 0.05% TFA

B: acetonitrile containing 0.05% TFA

Operating time: for 10 min.

Gradient elution procedure

Time (min)	Flow rate of flow	%A	%B
				0.0	30	90	10
5.0	30	90	10
				25	30	20	80
35	30	80	80

Solvents used for sample preparation: methanol, acetonitrile/methanol (1:1)

Method 003P

Column: YMCODA-A (500 mm. times.30 mm. times.10 μ)

Flow rate: 30mL/min

Injection volume: 5mL

Column heater temperature: ambient temperature

Wavelength: binary 220nm

Mobile phase:

a: water containing 0.05% TFA

B: acetonitrile containing 0.05% TFA

Operating time: for 10 min.

Gradient elution procedure

Time (min)	Flow rate of flow	%A	%B
				0.0	30	50	50
5.0	30	50	50
				25	30	100	100
35	30	100	100

Solvents used for sample preparation: methanol, acetonitrile/methanol (1: 1).

Method 004P

Column: YMCODA-A (500 mm. times.30 mm. times.10 μ)

Flow rate: 30mL/min

Injection volume: 5mL

Column heater temperature: ambient temperature

Wavelength: binary 220nm

Mobile phase:

a: water (W)

B: acetonitrile

Operating time: for 10 min.

Gradient elution procedure

Solvents used for sample preparation: methanol, acetonitrile/methanol (1: 1).

Example 1

Preparation of 1- (3- (3-aminopropyl) phenyl) -3-ethylpent-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3-ethylpent-1-yn-3-ol was prepared according to the method shown in scheme 1:

reaction scheme 1

Step 1: to CH containing 3- (3-bromophenyl) propionic acid (1) (25.0g,109.1mmol) ₂Cl₂To a stirred solution (150ml) was added oxalyl chloride (27.7g,218.3mmol) followed by DMF (2 drops). The solution was stirred at room temperature overnight. The resulting mixture was concentrated under reduced pressure to give a crude acid chloride, which was immediately used in the subsequent reaction.

Step 2: the crude product was dissolved in anhydrous THF (150ml) and cooled in an ice bath. Ammonia gas was bubbled through the solution for 3-4 minutes, and the mixture was warmed to room temperature and stirred overnight. The mixture was concentrated under reduced pressure. To the residue was added saturated NaHCO₃(100ml) and the mixture was extracted with EtOAc (2X 200 ml). With Na₂SO₄The combined organics were dried and concentrated under reduced pressure to give the amide as a white solid2. Yield (23.9g, 96%):¹HNMR(400MHz,DMSO-d₆)7.40(s,1H),7.35(dt,J=6.4,2.4Hz,1H),7.26(brs,1H),7.18-7.24(m,2H),6.75(brs,1H),2.78(t,J=7.6Hz,2H),2.34(t,J=7.6Hz,2H)。

and step 3: to an ice-cooled solution of amide 2(23.85g,104.6mmol) in anhydrous tetrahydrofuran was added BH₃THF (209ml of a 1.0M solution in THF, 209 mmol). The solution was warmed to room temperature and stirred for 18 hours. The reaction was quenched by slowly adding 6n hcl until pH 1. The solution was then stirred at room temperature for 4 hours, at which time the pH was adjusted by addition of 50% aqueous NaOH>10. The solution was extracted with EtOAc (2X 250 ml). The combined organic layers were washed with brine, over Na₂SO₄Drying and concentration under reduced pressure gave the crude amine, which was used immediately in the subsequent reaction.

And 4, step 4: crude 3- (3-bromophenyl) propan-1-amine (ca.104.6mmol) was stirred with ethyl trifluoroacetate (30ml) overnight. The mixture was concentrated under reduced pressure. Purification by flash column chromatography (20% EtOAc-hexanes) afforded trifluoroacetamide 3. Yield (21.1g, 62%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.43(s,1H),7.36(dt,J=7.2,2.0Hz,1H),7.19-7.25(m,2H),3.16(q,J=6.8Hz,2H),2.57(t,J=7.6Hz,2H),1.77(quint.,J=7.2Hz,2H)。

and 5: to a degassed solution of N- (3- (3-bromophenyl) propyl) -2,2, 2-trifluoroacetamide (3) (0.930g,3mmol) and 3-ethylpent-1-yn-3-ol (4) (0.670g,6mmol) in triethylamine (4mL) and DMF (12mL) was added PdCl₂(PPh₃)₂(0.053g,0.075mmol), tri-o-tolylphosphine (0.046g,0.15mmol), and CuI (0.014g,0.075 mmol). The resulting mixture was degassed and stirred at 90 ℃ under an argon atmosphere for 6 hours. The mixture was cooled to room temperature, then concentrated under reduced pressure and diluted with EtOAc (100mL) and water (70 mL). Delamination after vigorous shaking. The organic layer was treated with charcoal, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Purification by flash column chromatography (7-60% EtOAc-hexanes gradient elution) afforded N- (3- (3- (3-ethyl-3-hydroxypentyl) phenyl) propyl) -2,2, 2-trifluoroacetamide (5) as a yellow oil. Yield (0.663g, 65%):¹HNMR(400MHz,DMSO-d₆)9.40(s,1H),7.17-7.28(m,4H),5.11(s,1H),3.16(q,J=7.2Hz,2H),2.56(t,J=7.2Hz,2H),1.76(quint,J=7.2Hz,2H),1.53-1.67(m,4H),0.97(t,J=7.2Hz,6H)。

step 6: n- (3- (3- (3-ethyl-3-hydroxypentyl) phenyl) propyl) -2,2, 2-trifluoroacetamide (5) (0.660g,1.93mmol) is dissolved in MeOH (15mL) and K is added ₂CO₃(0.42g in 3mL of water, 3.0mmol) in water. The resulting mixture was stirred at 45 ℃ for 4 hours. After cooling, the reaction mixture was concentrated under reduced pressure and diluted with EtOAc (50mL) and water (50 mL). Delamination after vigorous shaking. The organic layer was dried (sodium sulfate), filtered, and concentrated under reduced pressure. By flash column chromatography (72:8:20 to 90:10:0 EtOAc/containing 7MNH₃MeOH/hexanes) gave example 1 as a clear oil. Yield (0.421g,89%):¹HNMR(400MHz,DMSO-d₆)7.15-7.26(m,4H),5.11(s,1H),2.56(t,J=7.6Hz,2H),2.47(t,J=5.2Hz,2H),1.55-1.65(m,6H),1.39(brs,2H),0.97(t,J=7.6Hz,6H)。

example 2

Preparation of 4- ((3- (3-aminopropyl) phenyl) ethynyl) heptan-4-ol

4- ((3- (3-aminopropyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure described in example 1.

Step 1: coupling of 4-ethynylhept-4-ol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide as a clear oil. Yield (0.103g, 31%):¹HNMR(400MHz,DMSO-d₆)9.40(s,1H),7.18-7.29(m,4H),3.16(q,J=7.2Hz,2H),2.56(t,J=7.2Hz,2H),1.76(quint,J=7.2Hz,2H),1.53-1.67(m,8H),0.97(t,J=7.2Hz,6H)。

step 2,2, 2-trifluoro-2To a solution of (E) -N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (0.1g,0.27mmol) in MeOH (3mL) was added concentrated NH₄OH (7mL) and the solution was stirred at room temperature overnight. The solvent was removed under reduced pressure and the residue was extracted twice with EtOAc. The combined organics were washed with water and brine, dried (sodium sulfate), and concentrated under reduced pressure to give example 2 as a clear oil. Yield (0.079g, 100%): ¹HNMR(400MHz,DMSO-d₆)7.14-7.26(m,4H),5.12(s,1H),5.11(s,1H),2.56(t,J=7.2Hz,2H),2.47(t,J=7.2Hz,2H),1.42-1.63(m,12H),0.90(t,J=7.2Hz,6H)。

Example 3

Preparation of 5- ((3- (3-aminopropyl) phenyl) ethynyl) nonan-5-ol

5- ((3- (3-aminopropyl) phenyl) ethynyl) nonan-5-ol was prepared according to the procedure described in example 1.

Step 1: coupling of 3-ethynylnonan-5-ol with bromide 3 gave N- (3- (3- (3-butyl-3-hydroxyhept-1-ynyl) phenyl) propyl) -2,2, 2-trifluoroacetamide as a tan oil. Yield (0.346g, 22%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.14-7.26(m,4H),5.11(s,1H),2.56(t,J=7.6Hz,2H),2.47(m,2H),1.43-1.62(m,14H),0.88(t,J=7.2Hz,6H)。

step 2: deprotection of N- (3- (3- (3-butyl-3-hydroxyhept-1-ynyl) phenyl) propyl) -2,2, 2-trifluoroacetamide affords example 3 as a light yellow oil. Yield (0.219g, 84%):¹HNMR(400MHz,DMSO-d₆)7.22-7.26(m,1H),7.14-7.17(m,3H),5.11(s,1H),2.56(t,J=7.6Hz,2H),2.49(t,J=6.8Hz,2H),1.25-1.62(m,14H),0.88(t,J=7.2Hz,6H)。

example 4

Preparation of 3- (3- (3-methoxy-3-propylhex-1-ynyl) phenyl) propan-1-amine

3- (3- (3-methoxy-3-propylhex-1-ynyl) phenyl) prop-1-amine was prepared according to the method used in example 1.

Step 1: coupling of 4-ethynyl-4-methoxyheptane with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- (3-methoxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.596g, 93%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.18-7.29(m,4H),3.25(s,3H),3.14-3.20(m,2H),2.56(t,J=7.6Hz,2H),1.73-1.80(m,2H),1.64(t,J=8.4Hz,4H),1.34-1.44(m,4H),0.88(t,J=7.2Hz,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-methoxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide gave example 4 as a clear oil. Yield (0.341g, 93%):¹HNMR(400MHz,DMSO-d₆)7.27-7.18(m,4H),3.25(s,3H),2.56(t,J=7.6Hz,2H),2.47(t,J=6.8Hz,2H),1.56-1.66(m,6H),1.32-1.44(m,6H),0.88(t,J=7.2Hz,6H)。

example 5

Preparation of 1- (3- (3-aminopropyl) phenyl) -3-methylhexan-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3-methylhexan-1-yn-3-ol was prepared according to the method used in example 1.

Step 1: coupling of 3-methylhexan-1-yn-3-ol with bromide 3 to give 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy) alkyne dimers contaminated with alkyne dimers3-methylhexan-1-ynyl) phenyl) propyl) acetamide. The yield (0.699g,>100%)：¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.25(dd,J=8.8,7.2Hz,1H),7.17-7.21(m,3H),5.29(s,1H),3.17(q,J=6.8Hz,2H),2.56(t,J=7.6Hz,2H),1.76(quint,J=7.2Hz,2H),1.48-1.61(m,4H),1.39(s,3H),0.90(t,J=7.6Hz,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-methylhexan-1-ynyl) phenyl) propyl) acetamide followed by flash column chromatography (72:8:20 to 90:10:0 EtOAc/7 MNH containing₃MeOH/hexanes) to afford example 5 as a yellow oil. Yield (0.371g,76%, two steps):¹HNMR(400MHz,DMSO-d₆)7.24(t,J=8Hz,1H),7.14-7.18(m,3H),5.29(brs,1H),2.56(t,J=7.6Hz,2H),2.47(t,J=7.2Hz,2H),1.41-1.62(m,6H),1.39(s,3H),1.34(brs,2H),0.90(t,J=7.6Hz,3H)。

example 6

Preparation of 1- (3- (3-aminopropyl) phenyl) -3, 5-dimethylhex-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3, 5-dimethylhex-1-yn-3-ol was prepared according to the method used in example 2.

Step 1: coupling of 3, 5-dimethylhex-1-yn-3-ol with bromide 3 (except for addition of the alkynol after degassing) as described in example 2 gave 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3, 5-dimethylhex-1-ynyl) phenyl) propyl) acetamide as a tan oil. Yield (0.287g, 40%);¹HNMR(400MHz,DMSO-d₆)9.41(brs,1H),7.26(t,J=7.6Hz,1H),7.16-7.20(m,3H),5.25(s,1H),3.16(q,J=6.8Hz,2H),2.56(t,J=7.2Hz,2H),1.90-1.96(m,1H),1.76(quint,J=7.6Hz,2H),1.53(m,2H),1.42(s,3H),0.96(d,J=6.8Hz,6H)。

step 2: except that the mixture is stirred at room temperature The mixture was allowed to stand overnight and 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3, 5-dimethylhex-1-ynyl) phenyl) propyl) acetamide was deprotected as in example 2 to give example 6 as a clear oil. Yield (0.141g, 72%):¹HNMR(400MHz,DMSO-d₆)7.14-7.27(m,4H),5.25(s,1H),2.56(t,J=7.2Hz,2H),2.47(t,J=6.0Hz,2H),1.93(quint,J=6.4Hz,1H),1.60(q,J=6.8Hz,2H),1.54(t,J=6.0Hz,2H),1.42(s,3H),1.35(brs,2H),0.97(d,J=6.4Hz,6H)。

example 7

Preparation of 4- (3- (3-aminopropyl) phenyl) -2-methylbut-3-yn-2-ol

4- (3- (3-aminopropyl) phenyl) -2-methylbut-3-yn-2-ol was prepared as described in example 2.

Step 1: coupling of 2-methylbut-3-yn-2-ol with bromide 3 in THF without tri-o-tolylphosphine gave 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-methylbut-1-ynyl) phenyl) propyl) acetamide. Yield (0.5g, 81%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.17-7.28(m,4H),3.16(q,J=7.2Hz,2H),2.56(t,J=7.2Hz,2H),1.76(quint,J=7.6Hz,2H),1.44(s,3H),1.35(s,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-methylbut-1-ynyl) phenyl) propyl) acetamide followed by flash chromatography (72:8:20 to 90:10:0 EtOAc/containing 7MNH₃MeOH/hexanes gradient elution) gave example 7 as a pale yellow oil. Yield (0.212g, 62%):¹HNMR(400MHz,DMSO-d₆)7.14-7.26(m,4H),5.41(brs,1H),2.56(t,J=7.2Hz,2H),2.47-2.50(m,2H),1.55-1.63(m,2H),1.44(s,6H),1.36(brs,2H)。

example 8

Preparation of 1- (3- (3-aminopropyl) phenyl) hex-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) hex-1-yn-3-ol was prepared according to the method described in example 7.

Step 1: hex-1-yn-3-ol was coupled with bromide 3 to give 2,2, 2-trifluoro-N- (3- (3- (3-hydroxyhex-1-ynyl) phenyl) propyl) acetamide as a tan oil. Yield (0.271g, 26%).

Step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxyhex-1-ynyl) phenyl) propyl) acetamide gave example 8 as a brown oil. Yield (0.086g, 45%):¹HNMR(400MHz,DMSO-d₆)7.16-7.26(m,4H),5.36(d,J=5.2Hz,1H),4.41(dt,J=6.4,5.2Hz,1H),2.56(t,J=7.2Hz,2H),2.47-2.49(m,2H),1.38-1.64(m,8H),0.90(t,J=7.2Hz,3H)。

example 9

Preparation of 3- (3- (3-methoxyprop-1-ynyl) phenyl) prop-1-amine

3- (3- (3-Methoxyprop-1-ynyl) phenyl) prop-1-amine was prepared according to the method described in example 7.

Step 1: coupling of 3-methoxyprop-1-yne with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- (3-methoxyprop-1-ynyl) phenyl) -propyl) acetamide as a pale yellow oil. Yield (0.193g, 32%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.21-7.31(m,4H),4.30(s,2H),3.31(s,3H),3.16(q,J=7.2Hz,2H),2.56(t,J=7.6Hz,2H),1.77(quint,J=7.2Hz,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-methoxyprop-1-ynyl) phenyl) propyl) acetamide gave example 9 as a clear oil. Yield (0.069g,54%):¹HNMR(400MHz,DMSO-d₆)7.19-7.28(m,4H),4.29(s,2H),3.31(s,3H),2.57(t,J=7.6Hz,2H),2.47(m,2H),1.56-1.63(m,2H),1.36(brs,2H)。

example 10

Preparation of 3- (3- (3-aminopropyl) phenyl) prop-2-yn-1-ol

3- (3- (3-aminopropyl) phenyl) prop-2-yn-1-ol was prepared as described in example 7.

Step 1: coupling of prop-2-yn-1-ol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- (3-hydroxyprop-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.148g, 26%):¹HNMR(400MHz,DMSO-d₆)9.41(brs,1H),7.19-7.29(m,4H),5.28(t,J=5.6Hz,1H),4.27(d,J=6.4Hz,2H),3.16(t,J=7.2Hz,2H),2.56(t,J=7.6Hz,2H),1.76(q,J=7.6Hz,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxyprop-1-ynyl) phenyl) propyl) acetamide provided example 10 as a clear oil. Yield (0.073g, 76%): ¹HNMR(400MHz,DMSO-d₆)7.17-7.27(m,4H),5.28(brs,1H),4.27(d,J=3.6Hz,2H),2.59(t,J=7.6Hz,2H),2.47(m,2H),1.52-1.63(m,4H)。

Example 11

Preparation of 1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclohexanol

1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclohexanol was prepared as described in example 7.

Step 1: coupling of 1-ethynylcyclohexanol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide as a tan oil. The yield (0.205g,>100%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.26(t,J=7.6Hz,1H),7.17-7.24(m,3H),5.37(s,1H),3.16(q,J=6.8Hz,2H),2.56(t,J=7.2Hz,2H),1.15-1.83(m,12H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide afforded example 11 as a light yellow solid. Yield (0.13g, 87%):¹HNMR(400MHz,DMSO-d₆)7.14-7.26(m,4H),5.37(s,1H),2.56(t,J=7.6Hz,2H),2.47(m,2H),1.15-1.83(m,14H)。

example 12

Preparation of 1- (3- (3-aminopropyl) phenyl) -3-tert-butyl-4, 4-dimethylpent-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3-tert-butyl-4, 4-dimethylpent-1-yn-3-ol was prepared as described in example 7.

Step 1: coupling of 3-tert-butyl-4, 4-dimethylpent-1-yn-3-ol with bromide 3 gives N- (3- (3- (3-tert-butyl-3-hydroxy-4, 4-dimethylpent-1-ynyl) phenyl) propyl) -2,2, 2-trifluoroacetamide as a tan oil. Yield (0.15g, 67%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.27(t,7.6Hz,1H),7.18-7.21(m,3H),4.92(s,1H),3.18(q,J=6.4Hz,2H),2.57(t,J=7.2Hz,2H),1.76(quint,J=7.6Hz,2H),1.15(brs,18H)。

step 2: except by flash chromatography (containing 10%7 MNH)₃MeOH-EtOAc) purified product, N- (3- (3- (3-tert-butyl-3-hydroxy-4, 4-dimethylpent-1-ynyl) phenyl) propyl) -2,2, 2-trifluoroacetamide was deprotected as described in example 7 to give example 12 as a yellow oil. Yield (0.102g, 90%): ¹HNMR(400MHz,DMSO-d₆)7.16-7.27(m,4H),4.92(s,1H),2.57(t,J=7.2Hz,2H),2.47(t,J=6.8Hz,2H),1.56(q,J=7.2Hz,2H),1.52(brs,2H),1.14(s,18H)。

Example 13

Preparation of 1- ((3- (3-aminopropyl) phenyl) ethynyl) -2,2,6, 6-tetramethylcyclohexanol

1- ((3- (3-aminopropyl) phenyl) ethynyl) -2,2,6, 6-tetramethylcyclohexanol was prepared as described in example 7.

Step 1: coupling of 1-ethynyl-2, 2,6, 6-tetramethylcyclohexanol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxy-2, 2,6, 6-tetramethylcyclohexyl) ethynyl) phenyl) propyl) acetamide as a light tan foam. Yield (0.192g, 84%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.27(t,J=7.6Hz,1H),7.18-7.23(m,3H),4.92(s,1H),3.18(q,J=6.8Hz,2H),2.57(t,J=7.2Hz,2H),1.76(quint,J=7.6Hz,2H),1.22-1.50(m,6H),1.14(s,6H),1.04(s,6H)。

step 2: except by flash chromatography (containing 10%7 MNH)₃MeOH-EtOAc) the product was purified, 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxy-2, 2,6, 6-tetramethylcyclohexyl) ethynyl) phenyl) propyl) acetamide was deprotected according to the procedure described in example 7. Example 13 was isolated as a white solid. Yield (0.016g, 73%):¹HNMR(400MHz,DMSO-d₆)7.15-7.27(m,4H),4.92(s,1H),2.57(t,J=7.2Hz,2H),2.47(t,J=7.2Hz,2H),1.26-1.66(m,10H),1.14(s,6H),1.04(s,6H)。

example 14

Preparation of (S) -1- (3- (3-aminopropyl) phenyl) oct-1-yn-3-ol

(S) -1- (3- (3-aminopropyl) phenyl) oct-1-yn-3-ol was prepared according to the method shown in reaction scheme 2.

Reaction formula 2

Step 1: to the oven dried flask was added N- (3- (3-bromophenyl) propyl) -2,2, 2-trifluoroacetamide (3) (0.507g,1.63mmol), (R) -octyn-3-ol (6) (0.33mL,2.26mmol), CuI (0.0090g,0.047mmol), PdCl ₂(PPh₃)₂(0.0430g,0.06mmol), diisopropylamine (0.34mL,2.4mmol) and anhydrous dioxane (2 mL). The flasks were alternately placed under vacuum and purged with argon, which was repeated 3 times. Adding P: (^tBu)₃(0.1mL,1.0M in dioxane, 0.1mmol), the flask was again placed under vacuum and then purged with argon. The mixture was heated at 45 ℃ for 17 hours under argon. The reaction mixture was diluted with EtOAc, filtered through a small pad of celite and silica gel, and concentrated under reduced pressure. Flash column chromatography (20 to 80% EtOAc-hexanes gradient elution) afforded alkyne 7 as a tan oil (0.215g, 37%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.18-7.28(m,4H),5.37(d,J=5.7,1H),4.39(dt,J=6.4,5.7Hz,1H),3.16(q,J=6.7Hz,2H),2.56(t,J=7.4Hz,2H),1.76(quint,J=7.3Hz,2H),1.58-1.65(m,2H),1.37-1.45(m,2H),1.24-1.30(m,4H),0.85(t,J=7.0Hz,3H)。

step 2: alkyne 7 (0.20)6g,0.58mmol) was dissolved in MeOH (15 mL). Addition of H₂O (1.5mL) and K₂CO₃(0.200g,1.45mmol), and the mixture was stirred at room temperature for 30 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in-10% MeOH-EtOAc, dried (sodium sulfate), filtered through a cotton plug, and then concentrated under reduced pressure. By flash chromatography (90 to 100% EtOAc-hexanes, then 10%7 MNH)₃MeOH-EtOAc solution) to afford example 14(0.154g, quantitative) as a light yellow oil.¹HNMR(400MHz,CD₃OD)。7.16-7.26(m,4H),4.49(t,J=6.8Hz,1H),2.61-2.67(m,4H),1.70-1.80(m,4H),1.50-1.53(m,2H),1.34-1.39(m,4H),0.93(t,J=7.0Hz,3H).ESIMSm/z242.27[M+H–H₂O]⁺。

Example 15

Preparation of (R) -1- (3- (3-aminopropyl) phenyl) oct-1-yn-3-ol

(R) -1- (3- (3-aminopropyl) phenyl) oct-1-yn-3-ol was prepared according to the method used in example 14.

Step 2: coupling of (R) -oct-1-yn-3-ol with bromide 3 gave (R) -2,2, 2-trifluoro-N- (3- (3- (3-hydroxyoct-1-ynyl) phenyl) propyl) acetamide as a tan oil. Yield (0.292g, 41%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.27(t,J=7.6Hz,1H),7.28-7.18(m,3H),5.36(d,J=5.6Hz,1H),4.40(q,J=5.6Hz,1H),3.16(q,J=6.8Hz,2H),2.56(t,J=7.2Hz,2H),1.76(q,J=7.6Hz,2H),1.16-1.65(m,8H),0.86(m,3H)。

step 2: except that the product was purified by flash chromatography (72:8:20 to 90:10:0 EtOAc/containing 7MNH₃MeOH/hexanes gradient elution) was purified, and (R) -2,2, 2-trifluoro-N- (3- (3- (3-hydroxyoct-1-ynyl) phenyl) propyl) acetamide was deprotected according to the procedure described in example 2 to afford example 15 as a pale yellow oil. Harvesting machineRate (0.119g, 56%):¹HNMR(400MHz,DMSO-d₆)7.16-7.27(m,4H),5.36(d,J=5.2Hz,1H),4.39(dt,J=5.2,6.4Hz,1H),2.56(t,J=7.2Hz,2H),2.47(tobs,J=6.8Hz,2H),1.55-1.65(m,4H),1.25-1.43(m,8H),0.86(t,J=6.8Hz,3H)。

example 16

Preparation of (R) -3- (3- (3-aminopropyl) phenyl) -1-phenylpropan-2-ynol

(R) -3- (3- (3-aminopropyl) phenyl) -1-phenylpropan-2-ynol was prepared according to the procedure described in example 15.

Step 1: coupling of N- (3- (3-bromophenyl) propyl) -2,2, 2-trifluoroacetamide (3) with (S) -3-phenylpropyn-3-ol gave (R) -2,2, 2-trifluoro-N- (3- (3-hydroxy-3-phenylpropan-1-ynyl) phenyl) propyl) acetamide (0.202 g; contaminated with alkyne dimers) as a tan oil. The product was used directly in the next synthesis step without purification.

Step 2: deprotection of (R) -2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-phenylprop-1-ynyl) phenyl) propyl) acetamide gave example 16 as a yellow oil which was purified by flash chromatography. Yield (0.079g, 53%): ¹HNMR(400MHz,CD₃OD)。7.56-7.59(m,2H),7.37(m,2H),7.25-7.32(m,3H),7.22(d,J=7.6Hz,1H),7.18(dt,J=7.2,2.0Hz,1H),5.61(s,1H),2.59-2.65(m,4H),1.75(quint,J=7.6Hz,2H).ESIMSm/z266.27[M+H]⁺。

Example 17

Preparation of 3- (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propan-1-amine

3- (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propan-1-amine was prepared according to the procedure shown in equation 3:

reaction formula 3

Step 1: to a degassed solution of 3- (3-bromophenyl) propan-1-ol (8) (0.95g,4.5mmol) and 2-methyl-3-butyn-2-ol (9) (1.6mL,16mmol) in triethylamine (25mL) was added PdCl₂(PPh₃)₃(0.095g,0.14mmol) and CuI (0.027g,0.14 mmol). The resulting solution was degassed and stirred at 70 ℃ for 15 hours under an argon atmosphere. The reaction mixture was concentrated under reduced pressure and diluted with EtOAc (50 mL). The solution was filtered through filter paper, washed with water (50mL) and brine (50mL), dried (sodium sulfate), and concentrated under reduced pressure. Purification by flash chromatography (10 to 100% EtOAc-hexanes gradient elution) afforded 4- (3- (3-hydroxypropyl) phenyl) -2-methylbut-3-yn-2-ol as a light tan oil (10); yield (0.78g, 80%):¹HNMR(400MHz,DMSO-d₆)7.29-7.18(m,4H),5.46(s,1H),4.48(t,J=5.2Hz,1H),3.38(q,J=6.0Hz,2H),2.59(t,J=7.2Hz,2H),1.66-1.73(m,2H),1.46(s,6H)。

step 2: to a solution of 4- (3- (3-hydroxypropyl) phenyl) -2-methylbut-3-yn-2-ol (10) (0.750g,3.4mmol) in toluene (50mL) was added KOH (0.390g,7mmol) in powder form. The resulting mixture was heated to reflux for 45 min, concentrated under reduced pressure to 10-15mL, and diluted with EtOAc (100 mL). The solution was washed with (2X 100mL) and brine (50mL), MgSO ₄Dried and concentrated under reduced pressure. Purification by flash chromatography gave 3- (3-ethynylphenyl) propan-1-ol (11) as a light tan oil. Yield (0.272g, 49%):¹HNMR(400MHz,DMSO-d₆)7.24-7.32(m,4H),4.49(t,J=5.2Hz,1H),4.15(s,1H),3.39(dt,J=6.4,5.6Hz,2H),2.60(t,J=7.6Hz,2H),1.66-1.73(m,2H)。

and step 3: to a gas-removed solution of 3- (3-ethynylphenyl) propan-1-ol (11) (0.270g,1.7mmol) and 2-iodo-1, 3-dimethylbenzene (0.392g,1.7mmol) in triethylamine (10mL) was added PdCl₂(PPh₃)₃(0.036g,0.05mmol) and CuI (0.010g,0.05 mmol). The resulting mixture was degassed and stirred at 70 ℃ under an argon atmosphere for 1.5 hours. The resulting reaction mixture was concentrated under reduced pressure and diluted with EtOAc (30 mL). The solution was filtered through filter paper, washed with water (2X 20mL) and washed with Na₂SO₄Dried above and concentrated under reduced pressure. Purification by flash chromatography (7 to 60% EtOAc-hexanes gradient elution) afforded (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propan-1-ol (12) as a light tan oil. Yield (0.085g, 19%). This material was used directly in the next synthetic step without further purification.

And 4, step 4: triphenylphosphine (0.087g,0.33mmol), phthalimide (0.0.49g,0.33mmol) and (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propan-1-ol (12) (0.085g,0.32mmol) were dissolved in anhydrous THF (3mL) under an argon atmosphere and the solution was cooled with an ice bath. Diethyl azodicarboxylate (0.052mL,0.33mmol) was added dropwise with rapid stirring, and the resulting mixture was stirred at room temperature for 2.5 hours. The reaction mixture was concentrated under reduced pressure and purified by flash chromatography (6 to 60% EtOAc-hexanes gradient elution) to give 2- (3- (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione (13) as a white solid. Yield (0.095g, 75%): ¹HNMR(400MHz,DMSO-d₆)7.82-7.89(m,4H),7.42(s,1H),7.13-7.35(m,6H),3.63(t,J=7.2Hz,2H),2.67(t,J=7.6Hz,2H),2.51(s,6H),1.90-1.97(m,2H)。

And 5: a solution of 2- (3- (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione (13) (0.094g,0.24mmol) and hydrazine hydrate (0.038g,0.75mmol) in dry EtOH (5mL) was heated at reflux for 3 hours. Additional hydrazine hydrate (0.038g,0.75mmol) was added and heating continued at reflux for 4 h. The solvent was removed under reduced pressure and the residue was taken up in hexane and Na₂SO₄Is subjected to sonication in the mixture of aqueous solutions. The mixture was filtered through celite and washed with hexane. In thatThe organic layer was concentrated under reduced pressure. By flash chromatography (10:1:9 EtOAc/containing 7MNH₃MeOH/hexane) to give 3- (3- ((2, 6-dimethylphenyl) ethynyl) phenyl) propan-1-amine as a colorless oil. Yield (0.015g,25%):¹HNMR(400MHz,DMSO-d₆)7.32-7.39(m,3H),7.25(m,1H),7.13-7.22(m,3H),2.65(t,J=7.6Hz,2H),2.54(t,J=7.2Hz,2H),2.46(s,6H),1.61-1.69(m,2H),1.40(brs,2H)。

example 18

Preparation of 4- ((3- (2-aminoethoxy) phenyl) ethynyl) heptan-4-ol

Preparation of 4- ((3- (2-aminoethoxy) phenyl) ethynyl) hept-4-ol according to the procedure shown in equation 4:

reaction formula 4

Step 1: to a solution of 3-bromophenol (14) (36.38g,210.3mmol) in acetone (175ml) was added K₂CO₃(0.033g,237mmol) and 2-bromoethanol (20ml,283.3 mmol). The mixture was heated to reflux under an argon atmosphere for 4 days and then cooled to room temperature. The solid was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was dissolved in diethyl ether (150mL) and washed with water (100mL), aqueous NaOH (10%,100mL, 3X 50mL,5%,200mL), water (100mL) and brine, MgSO ₄Drying and concentration under reduced pressure gave 2- (3-bromophenoxy) ethanol (15) as a light tan oil. Yield (21.07g, 46%):¹HNMR(400MHz,CDCl₃)7.14(t,J=7.8Hz,1H),7.07-7.12(m,2H),6.85(ddd,J=7.8,2.4,1.3Hz,1H),4.06(m,2H),3.95(m,2H),2.11(t,J=12.3Hz.1H)。

step 2: under argonTo a solution of 2- (3-bromophenoxy) ethanol (15) (16.06g,74.0mmol) and triethylamine (9.12g,90.13ml) in anhydrous CH under a gas atmosphere₂Cl₂(120ml) to an ice-cooled mixture was slowly added neat methanesulfonyl chloride (6ml,77.2mmol) and the reaction mixture was stirred at 0 ℃ for 15 minutes. A precipitate formed after the addition was complete. The mixture was concentrated under reduced pressure and the residue was dissolved in EtOAc, washed twice with water, once with brine and once with MgSO₄Dried and concentrated under reduced pressure. 2- (3-bromophenoxy) ethyl methanesulfonate (16) was isolated as a tan oil and used in the subsequent synthetic steps without further purification. Yield (21.32g, 98%):¹HNMR(400MHz,CDCl₃)7.16(t,J=7.8Hz,1H),7.11-7.14(m,1H),7.07(m,1H),6.39(ddd,J=7.6,2.5,1.8Hz,1H),4.56(m,2H),4.22(m,2H),3.08(s,3H)。

and step 3: to a solution of mesylate 16(24.05g,81.5mmol) in anhydrous DMF (160ml) was added potassium phthalimide (15.53g,83.8mmol) and the reaction mixture was stirred at 60 ℃ for 14 h. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was diluted with hexane-EtOAc (7:1,150ml) and water (150ml) and the mixture was shaken in a separatory funnel. The precipitate formed was removed by filtration, washed thoroughly with water and hexane, and then dried under vacuum to give N- (2- (3-bromophenoxy) ethyl) phthalimide (17) (22.05g,78%) as white, loose crystals. The organic layer of the filtrate was concentrated under reduced pressure and the residue was suspended in 10% EtOAc-hexanes. The solution was washed with water and the precipitate was collected by filtration, washed with water and hexane and dried under vacuum to give additional phthalimide 17(5.65 g). Combined yield (21.18g, 98%): ¹HNMR(400MHz,CDCl₃)7.86(m,2H),7.73(m,2H),7.03-7.12(m,3H),6.80(ddd,J=8.0,2.5and1.4Hz,1H),4.21(t,J=6.9Hz,2H),4.10(t,J=6.0Hz,2H)。

And 4, step 4: to a suspension of phthalimide 17(22.82g,65.9mmol) in anhydrous EtOH (200ml) was added hydrazine hydrate (6ml,123.7mmol) and the reaction mixture was heated to reflux under an argon atmosphere for 1.5 hours. After cooling to room temperature, the solid was removed by filtration and the filtrate was concentrated under reduced pressure. Recovering the residueSuspended in hexane (100ml) and the mixture filtered. The filtrate was concentrated under reduced pressure and then dried by concentration from EtOH then toluene to give amine 18 as a thick yellow oil. Yield (10.63g, 75%):¹HNMR(400MHz,CDCl₃)7.06-7.15(m,3H),6.84(ddd,J=8.0,2.5,1.2Hz,1H),3.96(t,J=5.3Hz,2H),3.07(t,J=5.1Hz,2H),1.43(s,2H)。

and 5: to a solution of amine 18(10.63g,49.2mmol) in dry THF (80ml) was added ethyl trifluoroacetate (12ml,100.6mmol), and the reaction mixture was stirred at room temperature overnight. The solution was concentrated under reduced pressure and the residue was dissolved in 50% EtOAc-hexanes. The solution was filtered through a silica gel layer and eluted with 50% EtOAc-hexane. Concentration under reduced pressure gave bromide 19 as a pale yellow oil which crystallized to a pale yellow solid upon standing. Yield (13.69g, 89%):¹HNMR(400MHz,CDCl₃)7.16(t,J=8.0Hz,1H),7.12-7.14(m,1H),7.05-7.07(m,1H),6.83(ddd,J=7.6,2.5,1.8Hz,1H),6.75(brs,1H),4.09(t,J=4.9Hz,2H),3.78(dt,J=5.5Hz,2H)。

step 6: with the exception of carrying out the reaction for 20 hours, coupling of bromide 19 with alkynol 20 following the procedure described in example 1 gave alkyne 21 as a yellow oil. Yield (0.89g, 73%): ¹HNMR(400MHz,CDCl₃)7.22(t,J=8.0Hz,1H),7.06(dt,J=7.6,1.0Hz,1H),6.93(dd,J=2.5,1.4Hz,1H),6.85(ddd,J=8.4,2.7,1.0Hz,1H),6.77(brs,1H),4.09(t,J=5.1Hz,2H),3.78(dt,J=5.5Hz,2H),2.00(s,1H),1.67-1.73(m,4H),1.57-1.61(m,4H),0.98(t,J=7.4Hz,6H)。

And 7: except that 5 equivalents of K are used at room temperature₂CO₃After 7 hours of reaction, alkyne 21 was deprotected according to the method described in example 1, followed by purification by flash column chromatography (9:1EtOAc (MeOH containing 7M aqueous ammonia) to give the trifluoroacetate of example 18 as an off-white solid, yield (5g, 76%):¹HNMR(400MHz,DMSO-d₆)7.23(t,J=7.8Hz,1H),6.92-6.93(m,1H),6.90-6.91(m,1H),6.85-6.86(m,1H),5.13(brs,1H),3.89(t,J=5.9Hz,2H),2.83(t,J=5.7Hz,2H),1.42-1.60(m,10H),0.89(t,J=7.2Hz,6H)；¹³CNMR(100MHz,DMSO-d₆)159.28,130.47,124.49,124.26,117,34,115.80,94.78,83,15,71.03,70.26,44.86,41.60,17.96,15.01.ESIMSm/z276.39[M+H]⁺,258.38[M+H-H₂O]⁺。

example 19

Preparation of 4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) heptan-4-ol

4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure shown in the following reaction scheme 5:

reaction formula 5

Step 1: lithium diisopropylamide (11mL of a 2M solution in THF, 22mmol) was added dropwise to a-78 deg.C solution of acetonitrile (1.05mL,20mmol) in dry THF (25mL) under an argon atmosphere. The resulting mixture was stirred at-78 ℃ for 15 minutes. 3-bromobenzaldehyde (22) (2.78g,15mmol) in dry THF (10mL) was added dropwise. The reaction mixture was warmed to room temperature, then concentrated under reduced pressure and diluted with EtOAc (75 mL). The solution was washed with water (50mL) and brine (50mL), Na₂SO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (20 to 100% EtOAc-hexanes gradient elution) afforded 3- (3-bromophenyl) -3-hydroxypropionitrile (23) as a pale yellow oil. Yield (2.75g, 81%): ¹HNMR(400MHz,DMSO-d₆)7.60(t,J=1.6Hz,1H),7.46(ddd,J=7.6,2.0,1.2Hz,1H),7.40(dd,J=7.6,2.0Hz,1H),7.31(t,J=7.6Hz,1H),6.05(d,J=4.8Hz,1H),4.87-4.92(m,1H),2.94-2.80(m,2H)。

Step 2: to 3- (3-bromophenyl) -3-hydroxypropionitrile (23) (2.70g,11.9mmol) in dry THF (20mL) under an argon atmosphere) Adding LiAlH into the ice-cooled solution₄Was added to the reaction solution (11.9mL of a 2M solution in THF, 23.8 mmol). The mixture was stirred at 0 ℃ for 45 minutes, diluted with ether (50mL) and saturated Na was added dropwise₂SO₄The reaction was quenched with aqueous solution (approximately 2 mL). Over MgSO₄After drying, the solution was filtered and concentrated under reduced pressure to give amine 24 as a pale green oil. Yield (2.30g, 84%). This material was used directly in the subsequent reaction without further purification.¹HNMR(400MHz,DMSO-d₆)7.49(m,1H),7.37(dt,J=7.2,1.6Hz,1H),7.23-7.31(m,2H),4.66(t,J=6.8Hz,1H),2.61(m,2H),1.61(q,J=6.8Hz,2H)。

And step 3: to a solution of 3-amino-1- (3-bromophenyl) propan-1-ol (24) (2.30g,10mmol) in anhydrous THF (20mL) was added ethyl trifluoroacetate (4.0mL,33.5 mmol). The reaction mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure. Purification by column chromatography (10 to 70% EtOAc-hexanes gradient elution) afforded N- (3- (3-bromophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide (25) as an oil containing-15% 2,2, 2-trifluoro-N- (3-hydroxy-3-phenylpropyl) acetamide. Yield (1.96g, 60%):¹HNMR(400MHz,DMSO-d₆)9.33(s,1H),7.51(t,J=2.0Hz,1H),7.41(dt,J=7.6,2.0Hz,1H),7.25-7.32(m,2H),5.46(d,J=6.4Hz,1H),4.55-4.60(m,1H),3.20-3.23(m,2H),1.75-1.82(m,2H)。

and 4, step 4: coupling of N- (3- (3-bromophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide (25) (1.95g,6mmol) with 4-ethynylhept-4-ol (20) as described in example 15 gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (26) as a light tan oil. Yield (0.87g, 37%): ¹HNMR(400MHz,DMSO-d₆)9.35(m,1H),7.29-7.34(m,3H),7.22-7.26(m,1H),5.39(d,J=4.4Hz,1H),5.12(s,1H),4.59(dt,J=8.4,4.8Hz,1H),3.25(quint,J=7.6Hz,2H),1.80(quint,J=8.0Hz,2H),1.44-1.63(m,8H),0.92(t,J=7.2Hz,6H)。

And 5: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (26) according to the procedure described in example 2 affords example 19. Yield (0.303g, 4)7%)：¹HNMR(400MHz,DMSO-d₆)7.16-7.32(m,4H),5.13(s,1H),4.65(t,J=6.0Hz,1H),2.56-2.64(m,2H),1.44-1.63(m,12H),0.90(t,J=7.6Hz,6H)。

Alternatively, example 19 can be prepared using the following reagents and conditions.

Step 1: to a cooled (-50 ℃ C.) stirred solution of potassium tert-butoxide (1M/THF,703mL,703mmol) under an argon atmosphere was added CH via syringe over 5 minutes₃CN (27.73g,675.6mmol) and the reaction mixture was stirred at-50 ℃ for 30 min. Then, a solution of 3-bromobenzaldehyde (22) (100g,540.5mmol) was added over 5 minutes. The reaction mixture was stirred at-50 ℃ for 30 minutes and warmed to room temperature. Addition of NH₄Aqueous Cl (25%,250mL), the mixture was stirred and the layers separated. The organic layer was washed with saturated brine (200mL), and anhydrous Na was added₂SO₄Drying, filtration, concentration of the filtrate under reduced pressure and drying of the residue under vacuum overnight gave the hydroxynitrile 23 as a pale yellow oil. Yield (117.6g, 96%):¹HNMR(400MHz,DMSO-d₆)7.60(t,J=1.6Hz,1H),7.46(ddd,J=7.6,2.0,1.2Hz,1H),7.40(dd,J=7.6,2.0Hz,1H),7.31(t,J=7.6Hz,1H),6.05(d,J=4.8Hz,1H),2.94-2.80(m,2H)。

step 2: to a solution of 3- (3-bromophenyl) -3-hydroxypropionitrile (23) (117.5g,519.8mmol) in dry THF (300mL) under an argon atmosphere was added borane-dimethylsulfide (68mL,675.7mmol) slowly over 30 minutes through a dropping funnel. The reaction mixture was boiled under reflux for 2.5 hours and cooled to room temperature. HCl solution (1.25M in EtOH, 350mL) was added slowly over 30 minutes, and the mixture was concentrated under reduced pressure. Water (400mL) was added, and the pH of the mixture was adjusted to 12 with aqueous NaOH (50% wt). By CH ₂Cl₂The product was extracted (500mL) with anhydrous Na₂SO₄The extract was dried and concentrated under reduced pressure to give hydroxylamine 24 as a colorless oil. Yield (104g, 87%):¹HNMR(400MHz,DMSO-d₆)7.49(m,1H),7.37(dt,J=7.2,1.6Hz,1H),7.23-7.31(m,2H),4.66(t,J=6.8Hz,1H),2.61(m,2H),1.61(q,J=6.8Hz,2H)。

step 3 to a cooled (0 ℃ C.) solution of MTBE in 3-amino-1- (3-bromophenyl) propan-1-ol (24) (40g,173.8mmol) was added ethyl trifluoroacetate (28mL,234.7mmol) over 7 minutes and the reaction mixture was stirred at room temperature for 50 minutes. Concentration under reduced pressure gave trifluoroacetamide 25 as a colorless oil. Yield (55.35g, 98%):

and 4, step 4: coupling of N- (3- (3-bromophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide (25) (55.35g,169.7mmol) with 4-ethynylhept-4-ol (20) (30.13g,214.9mmol) as described in example 1 gave crude 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (26) as a tan oil which was used directly in the next step without further purification. Yield (90.32g, quantitative):

and 5: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (26) according to the procedure described in example 14 was carried out by two column chromatographies on silica gel (first chromatography: EtOAc, then containing 10%7NNH ₃CH of/MeOH₂Cl₂(ii) a And (3) second chromatography: containing 8%7NNH₃CH of/MeOH₂Cl₂) After purification example 19 was obtained. Yield (29.97g, 60%):¹HNMR(400MHz,DMSO-d₆)7.16-7.32(m,4H),5.13(s,1H),4.65(t,J=6.0Hz,1H),2.56-2.64(m,2H),1.44-1.63(m,12H),0.90(t,J=7.6Hz,6H)。

example 20

Preparation of 4- ((3- (3-amino-2, 2-dimethylpropyl) phenyl) ethynyl) hept-4-ol

4- ((3- (3-amino-2, 2-dimethylpropyl) phenyl) ethynyl) hept-4-ol was prepared according to the procedure shown in equation 6:

reaction formula 6

Step 1: isobutyronitrile (2.15mL,24.0mmol) and anhydrous THF (60mL) were added to the oven dried flask under an argon atmosphere and cooled to-78 ℃. Lithium diisopropylamide (12mL of a 2.0M solution of heptane/THF/ethylbenzene, 24mmol) was added in aliquots over 20 minutes, and the reaction was stirred for 25 minutes. 3-bromobenzylbromide (27) (3.98g,15.92mmol) was added and the cold bath was removed. After stirring for a further 2 hours, the reaction was quenched by slow addition of water, followed by addition of EtOAc. The aqueous layer was partially saturated with sodium chloride. The layers were separated and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with water and brine, and Na₂SO₄Drying and concentration under reduced pressure gave nitrile 28 as an orange oil which solidified later (4.16g, quantitative, yield). This material was used directly in the subsequent synthetic steps without further purification. ¹HNMR(400MHz,CDCl₃)7.40-7.45(m,2H),7.20-7.25(m,2H),2.78(s,2H),1.36(s,6H)。

Step 2: to an ice-cooled solution of crude 3- (3-bromophenyl) -2, 2-dimethylpropionitrile (28) (3.0g,12.6mmol) in anhydrous THF (20mL) was slowly added BH₃THF (20mL of a 1M solution in THF, 20 mmol). The reaction was heated slowly and stirred for 19 hours. The reaction was quenched with dropwise addition of 6M HCl and then stirred for 1.5 hours. Volatiles were removed under reduced pressure. The aqueous layer was extracted twice with ether, then EtOAc was added and the mixture was made basic with 5M aqueous KOH. The layers were separated and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with brine, Na₂SO₄Drying and concentration under reduced pressure gave 3- (3-bromophenyl) -2, 2-dimethylpropan-1-amine (2.3g) as a pale yellow oil. This material was used directly in the subsequent step without further purification.¹HNMR(400MHz,CDCl₃),7.32-7.35(m,1H),7.30(t,J=1.7Hz,1H),7.13(t,J=7.7Hz,1H),7.06(dd,J=7.6,1.2Hz,1H),2.50(s,2H),2.47(s,2H),0.84(s,6H)。

And step 3: reacting the crude 3- (3-bromophenyl) -2,2-dimethylpropan-1-amine (2.3g) dissolved in THF (40 mL). Di-tert-butyl dicarbonate (2.3g,10.5mmol) and then triethylamine (2.8mL,20.1mmol) were added, and the mixture was stirred for 1.5 hours. The reaction mixture was concentrated under reduced pressure and the product was purified by flash chromatography (0-35% EtOAc-hexanes gradient elution) to give aryl bromide 29 as a colorless oil. Yield (3.3g, 77%):¹HNMR(400MHz,CDCl₃)　.7.34(d,J=7.6Hz,1H),7.27(t,J=1.6Hz,1H),7.14(t,J=7.7Hz,1H),7.05(d,J=7.8Hz,1H),4.58(brs,1H),2.98(d,J=6.5Hz,2H),2.48(s,2H),1.45(s,9H),0.85(s,6H)。

and 4, step 4: tert-butyl 3- (3-bromophenyl) -2, 2-dimethylpropylcarbamate (29) (3.2g,9.35mmol) was dissolved in EtOAc (55mL) and HCl-EtOAc (. about.4.2M, 20mL,84mmol) solution was added. The reaction was vented with a needle and stirred at room temperature for 2.5 hours. The reaction was then diluted with hexane and a white solid was collected on a fritted glass funnel. The mother liquor was concentrated under reduced pressure and suspended in 5-10% EtOAc-hexanes and the white solid was collected and combined with the first batch. The solid was dried overnight in a vacuum oven at room temperature to give pure 3- (3-bromophenyl) -2, 2-dimethylpropan-1-amine hydrochloride as a white solid. Yield (1.52g): ¹HNMR(400MHz,CDCl₃),8.53(brs,2H),7.37(dq,J=1.2and8.0Hz,1H),7.31(t,J=1.6Hz,1H),7.13(t,J=7.7Hz,1H),7.08(dt,J=8.0,1.6Hz,1H),2.83-2.84(m,2H),2.67(s,2H),1.09(s,6H)。

And 5: 3- (3-bromophenyl) -2, 2-dimethylpropan-1-amine hydrochloride (1.52g,5.45mmol) was dissolved in anhydrous THF (50 mL). Et was added slowly₃N (1.5mL,10.76mmol) to prepare a white slurry. Ethyl trifluoroacetate (2mL,16.8mmol) was added and the mixture was stirred at room temperature for 15.5 h. Additional ethyl trifluoroacetate (. about.0.75 mL,6.2mmol) and triethylamine (0.75mL,5.4mmol) were added and the mixture was stirred for 4 hours. The reaction mixture was concentrated under reduced pressure. The product was dissolved in EtOAc and washed with saturated NaHCO₃(2X) aqueous solution and brine, washing the solution with Na₂SO₄Drying and concentration under reduced pressure gave N- (3- (3-bromophenyl) -2, 2-dimethylpropyl) -2,2, 2-trifluoroacetamide (30) as a yellow oil. Yield (1.84g,58% yield, two steps run):¹HNMR(400MHz,CDCl₃),7.39(ddd,J=8.0,2.0,0.8Hz,1H),7.29(t,J=1.6Hz,1H),7.17(t,J=7.6Hz,1H),7.05(dt,J=7.6,1.6Hz,1H),6.16(brs,1H),3.24(d,J=6.8Hz,2H),2.53(s,2H),0.93(s,6H)。

Step 6: n- (3- (3-bromophenyl) -2, 2-dimethylpropyl) -2,2, 2-trifluoroacetamide (30) (0.489g,1.45mmol) was coupled with 4-ethynylhept-4-ol (20) (0.28g,2.0mmol) as described in example 15 and the product was purified by flash chromatography (0 to 50% EtOAc-hexanes gradient elution) to give 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -2, 2-dimethylpropyl) acetamide (31) as a yellow oil. Yield (0.350g, 61%): ¹HNMR(400MHz,CD₃OD),7.20-7.25(m,3H),7.12-7.15(m,1H),3.19(s,2H),2.54(s,2H),1.58-1.71(m,8H),0.98(t,J=7.2Hz,6H),0.85(s,6H)。

And 7: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -2, 2-dimethylpropyl) acetamide (31) (0.345g,0.87mmol) as described in example 1 and by flash chromatography (90 to 100% EtOAc-hexane then 10%3.5MNH₃MeOH-EtOAc) the product was purified to give example 20 as an oil, while the starting material was recovered. Yield (0.0847g,32% yield):¹HNMR(400MHz,CD₃OD)7.19-7.24(m,3H),7.11-7.13(m,1H),2.53(s,2H),2.44(s,2H),1.56-1.72(m,8H),0.98(t,J=7.2Hz,6H),0.85(s,6H)。

example 21

Preparation of 1- (3- (3-aminopropyl) phenyl) -3, 4-dimethylpent-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3, 4-dimethylpent-1-yn-3-ol was prepared according to the method used in example 1.

Step 1: so that the reaction conditions of the step 3,coupling of 4-dimethylpent-1-yn-3-ol with bromide 3 gives 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3, 4-dimethylpent-1-ynyl) phenyl) propyl) acetamide as a succinic oil. Yield (0.98g, 89%):¹HNMR(400MHz,CD₃OD)7.15-7.25(m,4H),3.27-3.31(m,2H),2.62(t,J=7.6Hz,2H),1.82-1.90(m,3H),1.50(s,3H),1.09(d,J=6.4Hz,3H),1.05(d,J=6.8Hz,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3, 4-dimethylpent-1-ynyl) phenyl) propyl) acetamide gave example 21 as a yellow oil. Yield (0.456g, 65%):¹HNMR(400MHz,CD₃OD)7.15-7.25(m,4H),2.60-2.65(m,4H),1.85(quint,J=6.8Hz,1H),1.72-1.79(m,2H),1.47(s,3H),1.09(d,J=6.8Hz,3H),1.05(d,J=6.8Hz,3H)。

example 22

Preparation of 4- (3- (3-aminopropyl) phenyl) -2-phenylbut-3-yn-2-ol

1- ((4- (3- (3-aminopropyl) phenyl) -2-phenylbut-3-yn-2-ol was prepared as described in example 7.

Step 1: coupling of 2-phenylbut-3-yn-2-ol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-phenylbut-1-ynyl) phenyl) propyl) acetamide as a yellow oil.¹HNMR(400MHz,DMSO-d₆)，9.41(brs,1H),7.62(m,2H),7.51(m,1H),7.36(m,2H),7.26(m,4H),6.15(s,1H),3.16(m,2H),2.57(m,2H),1.78(m,2H),1.69(s,3H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-phenylbut-1-ynyl) phenyl) propyl) acetamide gave example 22 as a yellow oil. Yield (0.122g,27% two steps):¹HNMR(400MHz,DMSO-d₆),7.60-7.63(m,1H),7.33-7.38(m,1H),7.18-7.28(m,7H),6.16(brs,1H),2.57(m,2H),2.51(m,2H),1.69(s,3H),1.56-1.63(m,2H),1.34(brs,2H)。

example 23

Preparation of 1- (3- (3-aminopropyl) phenyl) -4-methylpent-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -4-methylpent-1-yn-3-ol was prepared according to the method described in example 7.

Step 1: coupling of 4-methylpent-1-yn-3-ol with bromide 3 gives 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-4-methylpent-1-ynyl) phenyl) propyl) acetamide as a yellow oil contaminated with alkyne dimers, which is used directly in the subsequent reaction without purification.¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.18-7.29(m,4H),5.37(d,J=5.6Hz,1H),4.20(t,J=5.6Hz,1H),3.16(dt,J=6.8,6.0Hz,2H),2.56(t,J=7.6Hz,2H),1.70-1.81(m,3H),0.96(d,J=6.8Hz,3H),0.94(d,J=6.8Hz,3H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-4-methylpent-1-ynyl) phenyl) propyl) acetamide gave example 23 as a yellow oil. Yield (10.174g,47%, two steps):¹HNMR(400MHz,DMSO-d₆)7.15-7.27(m,4H),4.29(d,J=5.6Hz,1H),2.63(m,4H),1.88(m,1H),1.76(m,2H),0.96(d,J=6.8Hz,3H),0.94(d,J=6.4Hz,3H)。

example 24

Preparation of 1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclopentanol

1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclopentanol was prepared according to the method used in example 7.

Step 1: coupling of 1-ethynylcyclopentanol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) propyl) acetamide as a yellow oil, which was used directly in the subsequent reaction without purification:¹HNMR(400MHz,CD₃OD)7.15-7.25(m,4H),3.28(t,J=7.2Hz,2H),2.62(t,J=7.2Hz,2H),1.97-2.00(m,2H),1.73-1.91(m,8H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) propyl) acetamide afforded example 24 as a yellow oil. Yield (0.478g,62%, two steps):¹HNMR(400MHz,DMSO-d₆)7.14-7.34(m,4H),2.59-2.64(m,4H),1.97-2.00(m,4H),1.71-1.87(m,6H)。

example 25

Preparation of 1- (3- (3-aminopropyl) phenyl) -3,4, 4-trimethylpent-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3,4, 4-trimethylpent-1-yn-3-ol was prepared as described in example 1.

Step 1: coupling of 3,4, 4-trimethylpent-1-yn-3-ol with bromide 3 in a 1:1 mixture of DMF and triethylamine gave 2,2, 2-trifluoro-N- (3- (3-hydroxy-3, 4, 4-trimethylpent-1-ynyl) phenyl) propyl) acetamide as an orange oil. Yield (0.84g, 73%):¹HNMR(400MHz,CD₃OD)7.15-7.25(m,4H),3.29(t,J=7.2Hz,2H),2.61(t,J=8.0Hz,2H),1.86(quint,J=7.6Hz,2H),1.49(s,3H),1.09(brs,9H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3, 4, 4-trimethylpent-1-ynyl) phenyl) propyl) acetamide gave example 25 as a yellow oil. Yield (0.493g, 83%):¹HNMR(400MHz,DMSO-d₆)7.15-7.24(m,4H),2.60-2.65(m,4H),1.72-1.79(m,2H),1.49(s,3H),1.09(s,9H)。

example 26

Preparation of (S) -3- (3- (3-aminopropyl) phenyl) -1-phenylpropan-2-yn-1-ol

(S) -3- (3- (3-aminopropyl) phenyl) -1-phenylpropan-2-yn-1-ol was prepared according to the method described in example 1.

Step 1: coupling of (R) -1-phenylprop-2-yn-1-ol with bromide 3 gave (S) -2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-phenylprop-1-ynyl) phenyl) propyl) acetamide as an amber oil. Yield (0.73g, 62%):¹HNMR(400MHz,DMSO-d₆)7.57-7.59(m,2H),7.17-7.40(m,7H),5.60(s,1H),3.26-3.29(m,2H),2.62(t,J=7.6Hz,2H),1.86(quint,J=6.8Hz,2H)。

step 2: deprotection of (S) -2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-phenylprop-1-ynyl) phenyl) propyl) acetamide gave example 26 as a pale yellow oil. Yield (0.239g, 30%):¹HNMR(400MHz,CD₃OD)7.56-7.59(m,1H),7.16-7.39(m,8H),5.60(s,1H),2.58-2.62(m,4H),1.69-1.77(m,2H)。

example 27

Preparation of 1- (3- (2-aminoethoxy) phenyl) -3-ethylpent-1-yn-3-ol

1- (3- (2-Aminoethoxy) phenyl) -3-ethylpent-1-yn-3-ol was prepared as described in example 18.

Step 1: with the exception of allowing the reaction to proceed for 20 hours, 3-ethylpent-1-yn-3-ol was coupled with bromide 19 as described in example 18 to give N- (2- (3- (3-ethyl-3-hydroxypent-1-ynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide as a tan oil. Yield (0.52g, 60%):¹HNMR(400MHz,CDCl₃)7.22(t,J=8.0Hz,1H),7.06(dt,J=7.8,1.2Hz,1H),6.94(dd,J=2.5,1.4Hz,1H),6.89(brs,1H),6.85(ddd,J=8.2,2.5,1.0Hz,1H),4.08(t,J=5.1Hz,2H),3.76(dt,J=5.1Hz,2H),2.11(s,1H),1.76(m,4H),1.09(t,J=7.4Hz,6H)。

step 2: deprotection of N- (2- (3- (3-ethyl-3-hydroxypent-1-ynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide afforded example 27 as an oil which was solidified by standing in yield (0.243g, 65%):¹HNMR(400MHz,DMSO-d₆)7.24(t,J=8.0Hz,1H),6.90-6.94(m,2H),6.86-6.88(m,1H),5.13(brs,1H),3.89(t,J=5.9Hz,2H),2.83(t,J=5.8Hz,2H),1.54-1.65(m,4H),1.47(brs,2H),0.97(t,J=7.4Hz,6H).¹³CNMR(100MHz,DMSO-d₆)　.159.28,130.47,124.48,124.29,117.37,115.86,94.28,83.34,71.22,71.04,41.60,34.71,9.40.ESIMSm/z248.35[M+H]⁺,230.32[M+H-H₂O]⁺。

example 28

Preparation of 1- (3- (2-aminoethoxy) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol

1- (3- (2-Aminoethoxy) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol is prepared as described in example 18.

Step 1: 3-isopropyl-4-methylpent-1-yn-3-ol and bromide 19 were coupled as described in example 18, except that the reaction was allowed to proceed for 20 hours to give an oil2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-isopropyl-4-methylpent-1-ynyl) phenoxy) methyl) acetamide, which solidifies on standing. Yield (0.94g, 46%):¹HNMR(400MHz,CDCl₃)7.23(t,J=8.0Hz,1H),7.07(dt,J=7.6,1.0Hz,1H),6.95(dd,J=2.5,1.4Hz,1H),6.85(ddd,J=8.4,2.7,1.0Hz,1H),6.70(brs,1H),4.10(t,J=5.1Hz,2H),3.79(dt,J=5.1Hz,2H),2.04(m,2H),1.80(s,1H),1.09(d,J=6.7Hz,6H),1.05(d,J=6.7Hz,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-isopropyl-4-methylpent-1-ynyl) phenoxy) ethyl) acetamide provided example 28 as a white solid. Yield (0.529g, 76%):¹HNMR(400MHz,DMSO-d₆)　7.24(t,J=7.8Hz,1H),6.90-6.95(m,2H),6.87-6.88(m,1H),4.83(brs,1H),3.89(t,J=5.7Hz,2H),2.83(t,J=5.7Hz,2H),1.86(m,2H),1.47(brs,2H),0.98(d,J=6.8Hz,6H),0.93(d,J=6.7Hz,6H).¹³CNMR(100MHz,DMSO-d₆)　159.29,130.48,124.58,124.32,117.46,115.72,92.60,84.54,76.74,71.04,41.62,34.95,18.98,17.21.ESIMSm/z276.39[M+H]⁺,258.37[M+H-H₂O]⁺。

example 29

Preparation of 5- ((3- (2-aminoethoxy) phenyl) ethynyl) nonan-5-ol

5- ((3- (2-Aminoethoxy) phenyl) ethynyl) nonan-5-ol is prepared according to the procedure described for example 18.

Step 1: with the exception of carrying out the reaction for 18 hours, 5-ethynylnonan-5-ol was coupled with bromide 19 as described in example 18 to give N- (2- (3- (3-butyl-3-hydroxyhept-1-ynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide. Yield (1.06g, 75%):¹HNMR(400MHz,CDCl₃)7.23(t,J=8.0Hz,1H),7.06(dt,J=7.6and1.2Hz,1H),6.94(dd,J=2.5,1.4Hz,1H),6.86(ddd,J=8.4,2.7,1.0Hz,1H),6.72(brs,1H),4.10(t,J=5.3Hz,2H),3.79(dt,J=5.3Hz,2H),1.96(s,1H),1.70-1.75(m,4H),1.50-1.58(m,4H),1.34-1.43(m,4H),0.94(t,J=7.2Hz,6H)。

step 2: deprotection of N- (2- (3- (3-butyl-3-hydroxyhept-1-ynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide affords example 29 as an oil which solidifies on standing. Yield (0.695g, 92%): ¹HNMR(400MHz,DMSO-d₆)7.24(t,J=7.8Hz,1H),6.92-6.93(m,1H),6.90-6.91(m,1H),6.85-6.86(m,1H),5.13(brs,1H),3.89(t,J=5.7Hz,2H),2.83(t,J=5.7Hz,2H),1.52-1.60(m,6H),1.40-1.49(m,4H),1.25-1.34(m,4H),0.88(t,J=7.2Hz,6H).¹³CNMR(100MHz,DMSO-d₆)159.28,130.49,124.50,124.26,117.35,115.76,94.87,83.08,71.03,70.27,42.19,41.60,26.85,23.15,14.74.ESIMSm/z304.42[M+H]⁺,286.42[M+H-H₂O]⁺。

Example 30

Preparation of 4- (3- (2-aminoethoxy) phenyl) -2-methylbut-3-yn-2-ol

4- (3- (2-Aminoethoxy) phenyl) -2-methylbut-3-yn-2-ol was prepared according to the method described in example 18.

Step 1: coupling of 2-methylbut-3-yn-2-ol with bromide 3 gave 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-methylbut-1-ynyl) phenoxy) ethyl) acetamide by the procedure described in example 18, except that the reaction was allowed to proceed for 19 hours. Yield (0.667g, 70%):¹HNMR(400MHz,CDCl₃)7.23(t,J=7.8Hz,1H),7.06(dt,J=7.6and1.2Hz,1H),6.94(dd,J=2.5,1.4Hz,1H),6.86(ddd,J=8.2,2.5,1.0Hz,1H),6.74(brs,1H),4.09(t,J=4.9Hz,2H),3.80(dt,J=5.5Hz,2H),2.04(s,1H),1.61(s,6H)。

step 2: reacting 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy)Deprotection of the yl-3-methylbut-1-ynyl) phenoxy) ethyl) acetamide gave example 30 as a white solid. Yield (0.240g, 52%):¹HNMR(400MHz,DMSO-d₆)7.23(t,J=8.0Hz,1H),6.89-6.93(m,2H),6.86-6.88(m,1H),5.43(brs,1H),3.89(t,J=5.9Hz,2H),2.83(t,J=5.9Hz,2H),1.45(brs,2H),1.44(s,6H).¹³CNMR(100MHz,DMSO-d₆)　159.27,130.45,124.38,124.20,117.21,116.00,96.57,80.99,71.03,64.27,41.59,32.28.ESIMSm/z220.31[M+H]⁺,202.28[M+H-H₂O]⁺(ii) a HPLC (method A) t_R=2.79min。

Example 31

Preparation of 1- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclopentanol

1- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclopentanol was prepared according to the method described in example 18.

Step 1: coupling of 1-ethynylcyclopentanol with bromide 19 followed by the procedure described for example 18 gave 2,2, 2-trifluoro-N- (2- (3- ((1-hydroxycyclopentyl) ethynyl) phenoxy) ethyl) acetamide as a tan oil, except that the reaction was allowed to proceed for 19.5 hours. Yield (1.055g, 92%):¹HNMR(400MHz,CDCl₃)7.23(t,J=8.0Hz,1H),7.06(dt,J=7.6,1.2Hz,1H),6.95(dd,J=2.5,1.4Hz,1H),6.85(ddd,J=8.4,2.7,1.0Hz,1H),6.72(brs,1H),4.09(t,J=5.3Hz,2H),3.78(dt,J=5.1Hz,2H),2.00-2.09(m,4H),1.76-1.93(m,5H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- ((1-hydroxycyclopentyl) ethynyl) phenoxy) ethyl) acetamide gave example 31 as an oil which solidified upon standing. Yield (0.502g,66%): ¹HNMR(400MHz,DMSO-d₆)7.23(t,J=8.0Hz,1H),6.88-6.94(m,3H),5.28(brs,1H),3.89(t,J=5.7Hz,2H),2.83(t,J=5.7Hz,2H),1.82-1.89(m,4H),1.63-1.74(m,4H),1.48(brs,2H).¹³CNMR(100MHz,DMSO-d₆)159.27,130.45,124.50,124.18,117.20,115.93,95.65,81.97,73.44,71.01,42.66,41.58,23.75.ESIMSm/z246.33[M+H]⁺,228.30[M+H-H₂O]⁺(ii) a HPLC (method A) t_R=4.19min。

Example 32

Preparation of 1- (3- (3-aminopropyl) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol

1- (3- (3-aminopropyl) phenyl) -3-isopropyl-4-methylpent-1-yn-3-ol was prepared according to the method used in example 1.

Step 1: coupling of 3-isopropyl-4-methylpent-1-yn-3-ol with bromide 3 as described in example 17 gave 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-isopropyl-4-methylpent-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (1.375g,66%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.26(t,J=7.6Hz,1H),7.17-7.22(m,3H),4.81(s,1H),3.17(q,J=6.8Hz,2H),2.56(t,J=8.0Hz,2H),1.86(quint,J=6.8Hz,2H),1.76(quint,J=7.6Hz,2H),0.99(d,J=6.8Hz,6H),0.94(d,J=6.8Hz,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-isopropyl-4-methylpent-1-ynyl) phenyl) propyl) acetamide followed by flash chromatography (9:1 CH)₂Cl₂:7MNH₃MeOH) gave example 32 as a clear oil. Yield (0.835g,82%):¹HNMR(400MHz,DMSO-d₆)7.15-7.26(m,4H),4.82(brs,1H),2.56(t,J=7.6Hz,2H),2.47-2.52(m,2H),1.86(quint,J=6.8Hz,2H),1.59(quint,J=6.8Hz,2H),1.56(br.s,2H),1.05(d,J=6.8Hz,6H),1.03(d,J=6.8Hz,6H)。

example 33

Preparation of 4- ((3- (3-aminopropyl) phenyl) ethynyl) -2, 6-dimethylhept-4-ol

4- ((3- (3-aminopropyl) phenyl) ethynyl) -2, 6-dimethylheptan-4-ol was prepared according to the procedure described in example 32.

Step 1: coupling of 4-ethynyl-2, 6-dimethylhept-4-ol with bromide 3 gave 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-isobutyl-5-methylhexan-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (1.25g, 63%): ¹HNMR(400MHz,DMSO-d₆),9.40(brs,1H),7.14-7.28(m,4H),5.02(s,1H),3.17(q,J=6.8Hz,2H),2.56(t,J=7.6Hz,2H),1.93-1.99(m,2H),1.75(quint,J=7.6Hz,2H),1.47-1.56(m,4H),0.86-0.98(m,12H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-isobutyl-5-methylhexan-1-ynyl) phenyl) propyl) acetamide gave example 33 as a clear oil. Yield (0.73g, 77%):¹HNMR(400MHz,DMSO-d₆)7.22-7.26(m,1H),7.12-7.18(m,3H),5.04(brs,1H),2.56(t,J=7.2Hz,2H),2.50(t,J=6.8Hz,2H),1.91-2.01(m,2H),1.47-1.62(m,6H),0.98(m,6H),0.96(m,6H)。

example 34

Preparation of 4- (3- (3-aminopropyl) phenyl) but-3-yn-1-ol

4- (3- (3-aminopropyl) phenyl) but-3-yn-1-ol was prepared according to the method described for example 14.

Step 1: at room temperatureCoupling of but-3-yn-1-ol with bromide 3 gives 2,2, 2-trifluoro-N- (3- (3- (4-hydroxybut-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.9g, 62%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),7.15-7.26(m,4H),4.86(brs,1H),3.56(tapp,J=6.8Hz,2H),3.16(q,J=6.8Hz,2H),2.47-2.56(m,4H),1.76(quint,J=7.6Hz,2H)。

step 2: except by flash Chromatography (CH)₂Cl₂/EtOH/NH₄OH85:14:1) purification of the product example 34 was obtained as a clear oil by deprotecting 2,2, 2-trifluoro-N- (3- (3- (4-hydroxybut-1-ynyl) phenyl) propyl) acetamide as used in example 2. Yield (0.236g,65%):¹HNMR(400MHz,DMSO-d₆)7.12-7.24(m,4H),3.56(t,J=6.9Hz,2H),2.47-2.57(m,6H),1.59(quint,J=6.9Hz,2H)。

example 35

Preparation of 5- (3- (3-aminopropyl) phenyl) pent-4-yn-2-ol

5- (3- (3-aminopropyl) phenyl) pent-4-yn-2-ol was prepared according to the method described in examples 14 and 34.

Step 1: coupling of pent-4-yn-2-ol with bromide 3 at room temperature gave 2,2, 2-trifluoro-N- (3- (3- (4-hydroxypent-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.95g, 63%): ¹HNMR(400MHz,DMSO-d₆)　9.40(brs,1H),7.14-7.26(m,4H),4.80(s,1H),3.81(q,J=5.6Hz,1H),3.16(q,J=6.8Hz,2H),2.54(t,J=5.6Hz,2H),2.39(dd,J=16.8,6.8Hz,2H),1.76(quint,J=7.2Hz,2H),1.17(d,J=5.6Hz,3H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (4-hydroxypent-1-ynyl) phenyl) propyl) acetamide as described in example 34 gave example 35 as a clear oil. Yield (0.34g,94%):¹HNMR(400MHz,CDCl₃)7.24-7.25(m,1H),7.23(t,J=1.6Hz,1H),7.20(ddd,J=7.4,7.4,0.6Hz,1H),7.11(dt,J=7.2,1.6Hz,1H),4.04(dq,J=12.5,6.3Hz,1H),2.72(t,J=6.9Hz,2H),2.51-2.64(m,4H),1.72-1.79(m,2H),1.65(brs,3H),1.32(d,J=6.3Hz,3H)。

example 36

Preparation of 3- (3- ((2-methoxyphenyl) ethynyl) phenyl) propan-1-amine

3- (3- ((2-methoxyphenyl) ethynyl) phenyl) propan-1-amine was prepared according to the procedure shown in equation 7:

reaction formula 7

Step 1: alcohol 8 was coupled with phthalimide following the procedure described in example 17, except that diisopropyl azodicarboxylate was used instead of diethyl azodicarboxylate, to give phthalimide 32. Yield (6.9g, 50%):¹HNMR(400MHz,DMSO-d₆)7.78-7.86(m,4H),7.40-7.43(m,1H),7.29(dt,J=2.0,6.8Hz,1H),7.16-7.22(m,2H),3.58(t,J=6.8Hz,2H),2.61(t,J=7.2Hz,2H),1.84-1.93(m,2H)。

step 2: deprotection of phthalimide 32 following the procedure described in example 17 affords amine 33. Yield (4.2g,97%).

And step 3: protection of amine 33 with Boc anhydride following the procedure described in example 20 gave carbamate 34. Yield (5.57g,86%):¹HNMR(400MHz,DMSO-d₆)7.33(s,1H),7.26-7.34(m,1H),7.08-7.20(m,2H),4.55(brs,1H),3.15(q,J=6Hz,2H),2.61(t,J=8.0Hz,2H),1.79(quint,J=7.6Hz,2H),1.44(s,9H)。

and 4, step 4: carbamate 34 was coupled with alkyne 35 as described in example 1 to give alkyne 36 as a tan oil. Yield (0.201g, 36%):¹HNMR(400MHz,CDCl₃)7.53(dd,J=7.2,2Hz,2H),7.42-7.47(m,1H),7.36-7.38(m,1H),7.20-7.26(m,2H),7.11(d,J=8.0Hz,1H),6.98(t,J=7.6Hz,1H),6.86(m,1H),3.87(s,3H),2.91(q,J=6.4Hz,2H),2.55(tobs,J=7.6Hz,2H),1.65(quint,J=7.2Hz,2H),1.38(s,9H)。

and 5: alkyne 36(0.200gm,0.54mmol) was dissolved in CH₂Cl₂(5mL) and a solution of HCl in dioxane (15mL, saturated solution) was added. The mixture was stirred at room temperature for 18 hours. The mixture was concentrated under reduced pressure, then triturated with hexane (25 mL. times.2) to give a solid, which was washed with diethyl ether to give example 36 hydrochloride as a cream solid. Yield (0.127gm, 76%): ¹HNMR(400MHz,CDCl₃)8.38(brs,3H),7.48(dd,J=7.6,1.6Hz,1H),7.38(m,2H),7.29(tobs,J=7.2Hz,1H),7.22(t,J=8.0Hz,1H),7.14(d,J=7.6Hz,1H),6.92(t,J=7.6Hz,1H),6.88(d,J=8.4Hz,1H),3.89(s,3H),3.00(t,J=7.6Hz,2H),2.72(t,J=7.6Hz,2H),2.10(quint,J=7.6Hz,2H)。

Example 37

Preparation of 3- (3- (phenylethynyl) phenyl) propan-1-amine

3- (3- (phenylethynyl) phenyl) propan-1-amine is prepared according to the procedure described in example 36.

Step 1: coupling of the phenylalkyne with bromide 34 gave tert-butyl 3- (3- (phenylethynyl) phenyl) propylcarbamate as a tan oil. Yield (0.32g, 50%):¹HNMR(400MHz,CDCl₃)7.53(dd,J=7.2,2Hz,2H),7.42-7.47(m,1H),7.36-7.38(m,1H),7.20-7.26(m,2H),7.11(d,J=8.0Hz,1H),6.98(t,J=7.6Hz,1H),6.86(m,1H),4.54(brs,1H),3.14-3.17(m,2H),2.63(quint,J=7.6Hz,2H),1.76-1.86(m,2H),1.38(s,9H)。

step 2: deprotection of tert-butyl 3- (3- (phenylethynyl) phenyl) propylcarbamate afforded example 37 hydrochloride as an off-white solid. Yield (0.19g,73%):¹HNMR(400MHz,DMSO-d₆)8.08(brs,2H),7.55-7.57(m,1H),7.21-7.46(m,6H),7.21-7.30(m,2H),2.77(q,J=7.6Hz,2H),2.66(q,J=7.6Hz,2H),1.82-1.93(m,2H)。

example 38

Preparation of 3- (3- (cyclopentylethynyl) phenyl) propan-1-amine

3- (3- (cyclopentylethynyl) phenyl) propan-1-amine was prepared according to the method described in example 36.

Step 1: coupling of ethynylcyclopentane with bromide 34 gave t-butyl 3- (3- (cyclopentylethynyl) phenyl) propylcarbamate as a tan oil. Yield (0.70g,84%):¹HNMR(400MHz,DMSO-d₆)7.06-7.33(m,4H),2.85(quint,J=7.4Hz,1H),2.57-2.66(m,2H),2.62(t,J=8.0Hz,2H),1.93-2.01(m,2H),1.82(quint,J=7.6Hz,2H),1.66-1.75(m,2H),1.55-1.64(m,4H),1.45(m,9H)。

step 2: deprotection of tert-butyl 3- (3- (cyclopentylethynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 001P) afforded example 38 trifluoroacetate as a white solid. Yield (0.22g, 30%):¹HNMR(400MHz,DMSO-d₆)7.68(brs,3H),7.27(t,J=7.6Hz,1H),7.16-7.24(m,3H),2.85(quint,J=7.6Hz,1H),2.75(brs,2H),2.62(t,J=7.2Hz,2H),1.93-2.01(m,2H),1.82(quint,J=7.6Hz,2H),1.67-1.71(m,2H),1.56-1.66(m,4H)。

example 39

Preparation of 3- (3- (3-cyclopentylprop-1-ynyl) phenyl) prop-1-amine

3- (3- (3-Cyclopentylprop-1-ynyl) phenyl) prop-1-amine is prepared according to the method described in example 36.

Step 1: prop-2-ynyl cyclopentane was coupled with bromide 34 as used in example 36. Purification by flash chromatography (6% EtOAc-hexanes) afforded tert-butyl 3- (3- (cyclopentylethynyl) phenyl) propylcarbamate as a tan oil. Yield (0.70g, 84%):¹HNMR(400MHz,DMSO-d₆)7.06-7.33(m,4H),2.85(quint,J=7.4Hz,1H),2.57-2.66(m,2H),2.62(t,J=8.0Hz,2H),1.93-2.01(m,2H),1.82(quint,J=7.6Hz,2H),1.71(m,2H),1.59(m,4H),1.45(m,9H)。

step 2: deprotection of tert-butyl 3- (3- (3-cyclopentylprop-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 001P) afforded example 39 trifluoroacetamide as a white solid. Yield (0.4g,10%):¹HNMR(400MHz,DMSO-D₆)7.69(brs,2H),7.14-7.34(m,4H),2.76(t,J=6.4Hz,2H),2.62(t,J=7.6Hz,2H),2.42(d,J=6.8Hz,2H),2.08(m,1H),1.80(m,4H),1.48-1.70(m,4H),1.22-1.40(m,2H)。

example 40

Preparation of 3- (3- (3, 3-dimethylbut-1-ynyl) phenyl) propan-1-amine

3- (3- (3, 3-dimethylbut-1-ynyl) phenyl) propan-1-amine was prepared according to the procedure described in example 36.

Step 1: coupling of 3, 3-dimethylbut-1-yne with bromide 34 gave tert-butyl 3- (3- (3, 3-dimethylbut-1-ynyl) phenyl) propylcarbamate as a tan oil. Yield (0.43g, 54%).

Step 2: deprotection of tert-butyl 3- (3- (3, 3-dimethylbut-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 001P) gave example 40 trifluoroacetate as a pale yellow oil. Yield (0.08g, 18%):¹HNMR(400MHz,DMSO-d₆)7.79(brs,3H),7.27(t,J=7.6Hz,1H),7.16-7.22(m,3H),2.77(m,2H),2.61(t,J=7.6Hz,2H),1.82(quint,J=7.2Hz,2H),1.29(s,9H)。

EXAMPLE 41

Preparation of 3- (3- (cyclohexylethynyl) phenyl) propan-1-amine

3- (3- (cyclohexylethynyl) phenyl) propan-1-amine was prepared according to the procedure described in example 36.

Step 1: ethynylcyclohexane was coupled with bromide 34 as used in example 36. Purification by flash chromatography (5% EtOAc-hexanes) afforded tert-butyl 3- (3- (cyclohexylethynyl) phenyl) propylcarbamate as a tan oil. Yield (0.50g, 57%).

Step 2: tert-butyl 3- (3- (cyclohexylethynyl) phenyl) propylcarbamate, followed by purification by preparative HPLC (method 001P) gave example 41 trifluoroacetate as a cream solid. Yield (0.21g, 40%):¹HNMR(400MHz,DMSO-d₆)7.67(brs,3H),7.28(t,J=7.6Hz,1H),7.17-7.24(m,3H),2.74-2.79(m,1H),2.64(t,J=7.6Hz,4H),1.82(quint,J=7.2Hz,4H),1.67-1.68(m,2H),1.32-1.52(m,6H)。

example 42

Preparation of 3- (3- (3-phenylprop-1-ynyl) phenyl) propan-1-amine

3- (3- (3-phenylprop-1-ynyl) phenyl) prop-1-amine was prepared according to the method described in example 36.

Step 1: coupling of prop-2-ynylbenzene with bromide 34 afforded tert-butyl 3- (3- (3-phenylprop-1-ynyl) phenyl) propylcarbamate as a tan oil. Yield (0.85g,73%):¹HNMR(400MHz,CDCl₃)7.41-7.43(m,2H),7.35(t,J=8.0Hz,2H),7.10-7.28(m,5H),4.52(brs,1H),3.84(s,2H),3.14-3.16(m,2H),2.61(t,J=7.6Hz,2H),1.80(quint,J=7.6Hz,2H),1.48(s,9H)。

step 2: deprotection of tert-butyl 3- (3- (3-phenylprop-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method-001P) gave example 42 trifluoroacetate as a white solid. Yield (0.45g, 51%): ¹HNMR(400MHz,DMSO-d₆)7.69(brs,3H),7.35-7.42(m,4H),7.25-7.31(m,4H),7.20-7.22(m,1H),3.89(s,2H),2.76(t,J=7.2Hz,2H),2.63(t,J=7.6Hz,2H),1.82(quint,J=7.6Hz,2H)。

Example 43

Preparation of 3- (3- (pent-1-ynyl) phenyl) propan-1-amine

3- (3- (pent-1-ynyl) phenyl) propan-1-amine was prepared according to the method described in example 36.

Step 1: coupling pent-1-yne with bromide 34 to give tanTert-butyl 3- (3- (pent-1-ynyl) phenyl) propylcarbamate as a pale oil. Yield (0.35g, 58%):¹HNMR(400MHz,CDCl₃)7.07-7.33(m,4H),4.52(brs,1H),3.14-3.15(m,2H),2.58-2.66(m,2H),2.38(t,J=7.2Hz,2H),1.79(quint,J=7.6Hz,2H),1.64(q,J=7.2Hz,2H),1.45(s,9H),1.05(t,J=6.8Hz,3H)。

step 2: deprotection of tert-butyl 3- (3- (pent-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 001P) gave example 43 trifluoroacetate as a white solid. Yield (0.17g, 32%):¹HNMR(400MHz,DMSO-d₆)7.71(brs,3H),7.28(t,J=7.6Hz,1H),7.17-7.25(m,3H),2.76(t,J=7.2Hz,2H),2.62(t,J=7.2Hz,2H),2.39(t,J=6.8Hz,2H),1.82(quint,J=7.6Hz,2H),1.51-1.60(m,2H),1.00(t,J=7.6Hz,3H)。

example 44

Preparation of 3- (3- (hex-1-ynyl) phenyl) propan-1-amine

3- (3- (hex-1-ynyl) phenyl) prop-1-amine was prepared according to the method described in example 36.

Step 1: coupling of hex-1-yne with bromide 34 gave t-butyl 3- (3- (hex-1-ynyl) phenyl) propylcarbamate as a tan oil. Yield (0.64g, 64%).

Step 2: deprotection of tert-butyl 3- (3- (hex-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 004P) gave example 44 hydrochloride as a white solid. Yield (0.17g, 33%):¹HNMR(400MHz,DMSO-d₆)7.71(brs,3H),7.28(t,J=7.2Hz,1H),7.17-7.25(m,3H),2.76(t,J=7.6Hz,2H),2.62(t,J=7.2Hz,2H),2.42(t,J=7.0Hz,2H),1.82(quint,J=7.6Hz,2H),1.52(quint,J=7.0Hz,2H),1.44(quint,J=7.0Hz,2H),0.92(t,J=7.6Hz,3H)。

example 45

Preparation of 3- (3- (naphthalen-2-ylethynyl) phenyl) propan-1-amine

3- (3- (naphthalen-2-ylethynyl) phenyl) propan-1-amine was prepared according to the procedure described in scheme 8.

Reaction formula 8

Step 1: alcohol 11 was coupled with 2-bromonaphthalene (44) as described in example 17 to give alcohol 45. Yield (0.40g, 45%):¹HNMR(400MHz,CDCl₃)8.06(brs,1H),7.80-7.83(m,2H),7.58(dd,J=8.8,2.0Hz,1H),7.47-7.51(m,2H),7.41-7.43(m,2H),7.26-7.31(m,2H),7.18(d,J=7.6Hz,1H),4.77(brs,1H),4.11(t,J=6.4Hz,2H),2.71(t,J=7.6Hz,2H),1.99(quint,J=6.8Hz,2H)。

step 2: coupling of alcohol 45 with phthalimide following the procedure described in example 17 affords alkyne 46. Yield (0.40g, 80%):¹HNMR(400MHz,CDCl₃)8.44(d,J=8.0Hz,1H),7.82-7.88(m,4H),7.76(dd,J=7.2,1.2Hz,1H),7.70(dd,J=5.2,3.2Hz,2H),7.59-7.64(m,1H),7.52-7.56(m,1H),7.42-7.49(m,3H),7.29(d,J=7.6Hz,1H),7.22(d,J=8.0Hz,1H),3.79(t,J=7.2Hz,2H),2.73(t,J=7.2Hz,2H),2.09(quint,J=7.2Hz,2H)。

and step 3: deprotection of alkyne 46 following the procedure described in example 17 followed by purification by preparative HPLC (method 004P) afforded example 45 as a white solid. Yield (0.12g, 44%):¹HNMR(400MHz,DMSO-d₆)8.18(s,1H),7.80-7.95(m,3H),7.69(brs,2H),7.57-7.62(m,3H),7.45-7.49(m,2H),7.40(t,J=7.6Hz,1H),7.30(d,J=7.6Hz,1H),2.80(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),1.87(quint,J=7.6Hz,2H)。

example 46

Preparation of 3- (3- (diphenyl-3-ylethynyl) phenyl) propan-1-amine

3- (3- (diphenyl-3-ylethynyl) phenyl) propan-1-amine was prepared according to the procedure described in reaction formula 9.

Reaction formula 9

Step 1: alcohol 11 was coupled with 3-diphenylyne as described in example 1. Purification by flash chromatography (5% EtOAc-hexanes) afforded alcohol 47 as a tan oil. Yield (0.560g, 67%):¹HNMR(400MHz,CDCl₃)7.78(brs,1H),7.61(d,J=7.2Hz,2H),7.56(d,J=7.6Hz,1H),7.51(d,J=8.0Hz,1H),7.37-7.48(m,6H),7.29(d,J=7.6Hz,1H),7.19(d,J=7.6Hz,1H),3.70(dt,J=6.2,5.2Hz,2H),2.73(t,J=7.6Hz,2H),1.92(quint.,J=6.8Hz,2H),1.27(t,J=5.2Hz,1H)。

step 2: alcohol 47 was coupled with phthalimide according to the procedure described in example 17. Purification by flash chromatography (6% EtOAc-hexanes) afforded alkyne 48. Yield (0.320g, 42%):¹HNMR(400MHz,CDCl₃)7.84(dd,J=5.6,3.2Hz,2H),7.77(m,1H),7.71(dd,J=5.6,3.2Hz,2H),7.61-7.63(m,2H),7.32-7.57(m,8H),7.18-7.25(m,2H),3.77(t,J=7.2Hz,2H),2.70(t,J=7.2Hz,2H),2.02-2.09(m,2H)。

and step 3: deprotection of alkyne 48 following the procedure described in example 17 followed by purification by preparative HPLC (method 001P) afforded example 46 trifluoroacetate as a white sticky solid. Yield (0.16g, 52%): ¹HNMR(400MHz,DMSO-d₆)7.83(brs,1H),7.71-7.15(m,3H),7.67(brs,2H),7.38-7.55(m,8H),7.28-7.30(m,1H),2.77-2.82(m,2H),2.68(t,J=7.2Hz,2H),1.86(quint,J=7.6Hz,2H)。

Example 47

Preparation of 3-amino-1- (3- (cyclopentylethynyl) phenyl) propan-1-ol

3-amino-1- (3- (cyclopentylethynyl) phenyl) propan-1-ol was prepared according to the procedure described in reaction formula 10.

Reaction scheme 10

Step 1: the bromide 24 was coupled with di-tert-butyl dicarbonate as used in example 20. Purification by flash chromatography (13% EtOAc-hexanes) afforded tert-butyl 3- (3-bromophenyl) -3-hydroxypropyl carbamate (49) as a thick brown oil. Yield (4.0g, 48%):¹HNMR(400MHz,CDCl₃)7.53(s,1H),7.39(d,J=8.0Hz,1H),7.29(d,J=7.6Hz,1H),7.20(t,J=7.6Hz,1H),4.87(brs,1H),4.71(d,J=6.4Hz,1H),3.64(brs,1H),3.50-3.59(m,1H),3.12-3.19(m,1H),1.77-1.87(m,2H),1.46(s,9H)。

step 2: coupling of ethynylcyclopentane with tert-butyl 3- (3-bromophenyl) -3-hydroxypropylcarbamate (49) gave tert-butyl 3- (3- (cyclopentylethynyl) phenyl) -3-hydroxypropylcarbamate (50) as a tan oil. Yield (0.386g, 92%).

And step 3: deprotection of tert-butyl 3- (3- (cyclopentylethynyl) phenyl) -3-hydroxypropylcarbamate (50) followed by purification by preparative HPLC (method 001P) gave example 47 tris as a white solidA fluoroacetate. Yield (0.15g,37%):¹HNMR(400MHz,CDCl₃)7.88(brs,3H),7.17-7.31(m,4H),4.85(dd,J=7.6,4.0Hz,1H),3.11-3.17(m,2H),2.69(quint,J=7.2Hz,2H),1.56-2.02(m,10H)。

example 48

Preparation of 3-amino-1- (3- (3-cyclopentylprop-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (3-cyclopentylprop-1-ynyl) phenyl) propan-1-ol was prepared according to the method described in example 47.

Step 1: coupling of prop-2-ynylcyclopentane with tert-butyl 3- (3-bromophenyl) -3-hydroxypropylcarbamate (49) affords tert-butyl 3- (3- (3-cyclopentylprop-1-ynyl) phenyl) -3-hydroxypropylcarbamate as a tan oil. Yield (0.11g 26%).

Step 2: deprotection of tert-butyl 3- (3- (3-cyclopentylprop-1-ynyl) phenyl) -3-hydroxypropylcarbamate followed by purification by preparative HPLC (method 001P) afforded example 48 trifluoroacetate as a white solid. Yield (0.05g, 44%):¹HNMR(400MHz,DMSO-d₆)7.57(brs,3H),7.26-7.35(m,4H),4.67(dd,J=7.6,4.8Hz,1H),2.81-2.86(m,2H),2.09(quint,J=8.8Hz,2H),1.74-1.88(m,5H),1.52-1.65(m,4H),1.27-1.35(m,2H)。

example 49

Preparation of 3-amino-1- (3- (3-phenylprop-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (3-phenylprop-1-ynyl) phenyl) propan-1-ol was prepared according to the procedure described for example 47.

Step 1: coupling of prop-2-ynylbenzene with bromide 49 gave 3-hydroxy-3- (3- (3-phenylprop-1-ynyl) phenyl) propylcarbamic acid tert-butyl ester as a tan oil. Yield (0.404g, 91%):¹HNMR(400MHz,CDCl₃)7.40-7.45(m,3H),7.32-7.36(m,3H),7.20-7.29(m,3H),4.87(brs,1H),4.72(brs,1H),3.83(s,2H),3.51-3.54(m,1H),3.35(brs,1H),3.12-3.19(m,1H),1.81-1.84(m,2H),1.45(s,9H)。

step 2: deprotection of tert-butyl 3-hydroxy-3- (3- (3-phenylprop-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 001P) afforded example 49 trifluoroacetate as a white solid. Yield (0.114g, 27%):¹HNMR(400MHz,CDCl₃)7.92(brs,3H),7.26-7.37(m,5H),7.16-7.23(m,4H),4.79(dd,J=8.4,3.6Hz,1H),3.75(s,2H),3.02-3.16(m,2H),1.93-1.98(m,2H)。

example 50

Preparation of 6- (3- (3-amino-1-hydroxypropyl) phenyl) hex-5-yn-1-ol

6- (3- (3-amino-1-hydroxypropyl) phenyl) hex-5-yn-1-ol was prepared according to the method described in example 47.

Step 1: coupling of hex-5-yn-1-ol with bromide 49 gave 3-hydroxy-3- (3- (6-hydroxyhex-1-ynyl) phenyl) propylcarbamic acid tert-butyl ester as a tan oil. Yield (0.405g, 77%).

Step 2: deprotection of tert-butyl 3-hydroxy-3- (3- (6-hydroxyhex-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 004P) gave example 50 hydrochloride as a white solid. Yield (0.12g, 32%):¹HNMR(400MHz,DMSO-d₆)7.87(brs,3H),7.25-7.35(m,4H),5.51(brs,1H),4.68(dd,J=7.8,4.4Hz,1H),4.46(t,J=6.4Hz,1H),3.40-3.44(m,2H),2.77-2.88(m,2H),2.41-2.44(m,2H),1.80-1.93(m,2H),1.56-1.62(m,4H)。

example 51

Preparation of 4- (3- (3-amino-1-hydroxypropyl) phenyl) but-3-yn-1-ol

4- (3- (3-amino-1-hydroxypropyl) phenyl) but-3-yn-1-ol was prepared as described for example 47.

Step 1: coupling of but-3-yn-1-ol with bromide 49 gave 3-hydroxy-3- (3- (4-hydroxybut-1-ynyl) phenyl) propylcarbamic acid tert-butyl ester as a tan oil. Yield (0.27g, 56%):¹HNMR(400MHz,DMSO-d₆)7.42(s,1H),7.31(m,3H),5.60(s,1H),3.89(q,J=8.8Hz,2H),3.80(q,J=5.8Hz,2H),2.70(t,J=6.2Hz,2H),2.54(t,J=6.2Hz,2H),1.81(m,2H),1.49(s,9H)。

step 2: deprotection of tert-butyl 3-hydroxy-3- (3- (4-hydroxybut-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 004P) gave example 51 hydrochloride as a clear oil. Yield (0.03g, 8%):¹HNMR(400MHz,DMSO-D₆)7.64(brs,3H),7.26-7.36(m,4H),4.90(t,J=5.4Hz,1H),4.68(dd,J=7.7,4.6Hz,1H),3.58(dt,J=6.4,5.9Hz,2H),2.80-2.85(m,2H),2.57(m,2H),1.81-1.88(m,2H)。

example 52

Preparation of 3-amino-1- (3- (cyclohexylethynyl) phenyl) propan-1-ol

3-amino-1- (3- (cyclohexylethynyl) phenyl) propan-1-ol was prepared according to the procedure described for example 47.

Step 1: coupling of ethynylcyclohexane with bromide 49 gave tert-butyl 3- (3- (cyclohexylethynyl) phenyl) -3-hydroxypropylcarbamate as a tan oil. Yield (0.3g, 75%).

Step 2: deprotection of tert-butyl 3- (3- (cyclohexylethynyl) phenyl) -3-hydroxypropyl carbamate followed by purification by preparative HPLC (method 004P) gave example 52 hydrochloride as a white solid. Yield (0.05g, 16%):¹HNMR(400MHz,DMSO-d₆)8.01(brs,3H),7.23-7.34(m,4H),5.61(brs,1H),4.68(dd,J=8.0,4.4Hz,1H),2.77-2.87(m,2H),2.61-2.65(m,1H),1.78-1.93(m,4H),1.67-1.69(m,2H),1.42-1.51(m,3H),1.32-1.39(m,3H)。

example 53

Preparation of 3-amino-1- (3- (hept-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (hept-1-ynyl) phenyl) propan-1-ol was prepared according to the procedure described for example 47.

Step 1: coupling of hept-1-yne with bromide 49 gave tert-butyl 3- (3- (hept-1-ynyl) phenyl) -3-hydroxypropylcarbamate as a clear oil. Yield (0.32g, 60%).

Step 2: deprotection of tert-butyl 3- (3- (hept-1-ynyl) phenyl) -3-hydroxypropyl carbamate followed by purification by preparative HPLC (method 004P) gave example 53 hydrochloride as a white solid. Yield (0.03g, 11%):¹HNMR(400MHz,DMSO-d₆)7.63(brs,3H),7.24-7.33(m,4H),5.59(d,J=4.4Hz,1H),4.65-4.67(m,1H),2.82(brs,2H),2.40(t,J=7.2Hz,2H),1.78-1.85(m,2H),1.49-1.55(m,2H),1.28-1.40(m,4H),0.88(t,J=7.2Hz,3H)。

example 54

Preparation of 3-amino-1- (3- ((2-methoxyphenyl) ethynyl) phenyl) propan-1-ol

3-amino-1- (3- ((2-methoxyphenyl) ethynyl) phenyl) propan-1-ol was prepared according to the procedure described for example 47.

Step 1: coupling of 1-ethynyl-2-methoxybenzene with bromide 49 gave 3-hydroxy-3- (3- ((2-methoxyphenyl) ethynyl) phenyl) propylcarbamic acid tert-butyl ester as a tan oil. Yield (0.23g, 50%): ¹HNMR(400MHz,CDCl₃)7.58(s,1H),7.51(dd,J=7.6,6.0Hz,1H),7.46-7.49(m,1H),7.30-7.34(m,3H),6.97(dd,J=7.6,1.2Hz,1H),6.92(d,J=8.4,1H),4.88(brs,1H),4.76(quint\,J=4.4Hz,1H),3.93(s,3H),3.54(brs,1H),3.32(s,1H),3.18(ddd,J=14.4,10.8,5.2Hz,1H),1.84-1.88(m,2H),1.51(s,9H)。

Step 2: deprotection of tert-butyl 3-hydroxy-3- (3- ((2-methoxyphenyl) ethynyl) phenyl) propylcarbamate followed by purification by preparative HPLC (method 001P) afforded example 54 trifluoroacetate as a white solid. Yield (0.15g, 63%):¹HNMR(400MHz,DMSO-d₆)7.65(brs,3H),7.35-7.49(m,6H),7.10(d,J=8.0Hz,1H),6.98(t,J=7.6Hz,1H),5.67(m,1H),4.73(m,1H),3.86(s,3H),2.85(m,2H),1.83-1.91(m,2H)。

example 55

4- ((3- (2-aminoethoxy) phenyl) ethynyl) tetrahydro-2H-thiopyran-4-ol

4- ((3- (3-aminopropyl) phenyl) ethynyl) tetrahydro-2H-thiopyran-4-ol was prepared according to the procedure described for example 18.

Step 1: coupling of 4-ethynyltetrahydro-2H-thiopyran-4-ol with bromide 19 at 60 ℃ overnight in THF followed by purification by flash chromatography (EtOAc/heptane (2:1)) afforded 2,2, 2-trifluoro-N- (3- (3- ((4-hydroxytetrahydro-2H-thiopyran-4-yl) ethynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.822g, 72%):¹HNMR(400MHz,CDCl₃)7.26(t,J=4.5Hz,1H),7.07(dt,J=7.6,1.2Hz,1H),6.95(dd,J=2.5,1.4Hz,1H),6.87(ddd,J=8.2,2.5,0.8Hz,1H),4.10(m,3H),3.79(q,J=5.5Hz,2H),2.73-2.92(m,4H),2.26-2.31(m,2H),2.01-2.04(m,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- ((4-hydroxytetrahydro-2H-thiopyran-4-yl) ethynyl) phenyl) propyl) acetamide followed by flash Chromatography (CH)₂Cl₂/EtOH/NH₄OH85:14:1) gave example 55 as a white amorphous solid. Yield (0.41g,68%):¹HNMR(400MHz,DMSO-d₆) 7.23-7.28 (m,1H),6.93-6.98(m,3H),6.72(brs,1H),5.69(brs,1H),3.90(t, J =5.8Hz,2H),2.83(t, J =5.8Hz,2H),2.69(t, J =5.6Hz,4H),2.09(dt, J =13.0 and 4.6Hz,2H),1.80 (quant, J =6.6Hz,2H),1.60(brs, 2H).

Example 56

Preparation of 4- ((3- (2-aminoethoxy) phenyl) ethynyl) tetrahydro-2H-pyran-4-ol

4- ((3- (2-aminoethoxy) phenyl) ethynyl) tetrahydro-2H-pyran-4-ol was prepared according to the procedure described for example 55.

Step 1: coupling of 4-ethynyltetrahydro-2H-pyran-4-ol with bromide 19 gave 2,2, 2-trifluoro-N- (3- (3- ((4-hydroxytetrahydro-2H-pyran-4-yl) ethynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.12g, 22%).

Step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- ((4-hydroxytetrahydro-2H-pyran-4-yl) ethynyl) phenyl) propyl) acetamide provided example 56 as a white amorphous solid. Yield (0.050g, 57%):¹HNMR(400MHz,DMSO-d₆)7.23–7.27(m,1H),6.93-6.98(m,3H),5.70(brs,1H),3.90(t,J=5.8Hz,2H),3.72-3.78(m,2H),3.51-3.57(m,2H),2.83(t,J=5.8Hz,2H),1.82-1.97(m,2H),1.64-1.70(m,2H),1.52(brs,2H)。

example 57

Preparation of 1- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclohexanol

1- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclohexanol was prepared according to the procedure used in example 32.

Step 1: coupling of 1-ethynylcyclohexanol with bromide 17 gave 2- (2- (3- ((1-hydroxycyclohexyl) ethynyl) phenoxy) ethyl) isoindoline-1, 3-dione as a pale yellow oil. Yield (1.22g, 57%):¹HNMR(400MHz,DMSO-d₆)7.82-7.88(m,4H),7.21(t,J=8.0Hz,1H),6.92(dt,J=8.0,0.8Hz,1H),6.84-6.88(m,2H),5.38(bs,1H),4.20(t,J=5.6Hz,2H),3.95(t,J=5.6Hz,2H),1.78-1.82(m,2H),1.59-1.62(m,2H),1.41-1.53(m,5H),1.22-1.94(m,1H)。

step 2: deprotection of 2- (2- (3- ((1-hydroxycyclohexyl) ethynyl) phenoxy) ethyl) isoindoline-1, 3-dione as described in example 17 gave example 57 as a white amorphous solid, except that the reaction temperature was 70 ℃. Yield (0.47g, 67%): ¹HNMR(400MHz,DMSO-d₆)7.24(t,J=8.0Hz,1H),6.93(dt,J=8.0,1.2Hz,1H),6.88-6.91(m,2H),5.35(bs,1H),3.89(t,J=6.0Hz,2H),2.83(t,J=6.0Hz,2H),1.77-1.84(m,2H),1.45-1.63(m,9H),1.20-1.23(m,1H)。

Example 58

Preparation of 1- ((3- (3-aminopropyl) phenyl) ethynyl) cycloheptanol

1- ((3- (3-aminopropyl) phenyl) ethynyl) cycloheptanol was prepared according to the procedure described for example 32.

Step 1: coupling of 1-ethynylcycloheptanol with bromide 3 as used in example 17 gave 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxycycloheptyl) ethynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (1.78g, 60%):¹HNMR(400MHz,DMSO-d₆)　9.40(s,1H),7.26(t,J=7.6,1H),7.17-7.22(m,3H),5.26(s,1H),3.16(q,J=6.0Hz,2H),2.56(t,J=7.2Hz,2H),1.91-1.97(m,2H),1.73-1.79(m,4H),1.45-1.63(m,8H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- ((1-hydroxycycloheptyl) ethynyl) phenyl) propyl) acetamide following the procedure used in preparative example 1 gave example 58 as a clear oil. Yield (0.635g, 86%):¹HNMR(400MHz,DMSO-d₆)7.24(t,J=8.0Hz,1H),7.15-7.19(m,3H),5.28(brs,1H),2.56(t,J=7.2Hz,2H),2.44(m,2H),1.91-1.97(m,2H),1.73-1.79(m,2H),1.44-1.63(m,10H),1.32(brs,2H)。

example 59

Preparation of 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cycloheptanol

1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cycloheptanol was prepared according to the method described in example 32.

Step 1: coupling of 1-ethynylcycloheptanol with bromide 25 followed by purification by flash chromatography (2:1 to 3:2 to 1:1 hexanes/EtOAc) afforded 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((1-hydroxycycloheptyl) ethynyl) phenyl) propyl) acetamide as a tan oil. Yield (0.97g, 57%):¹HNMR(400MHz,CDCl₃)7.40(t,J=1.6Hz,1H),7.38(m,1H),7.28(dt,J=6.8,1.6Hz,1H),7.24-7.34(m,2H),4.84(ddd,J=6.4,5.2,3.2Hz,1H),3.66(dddd,J=12.0,7.2,6.8,6.8Hz,1H),3.64(dddd,J=12.4,8.0,5.6,4.8Hz,1H),2.56(d,J=2.4Hz,1H),2.10(dd,J=14.0,7.6Hz,2H),2.04(s,1H),1.86-2.00(m,4H),1.56-1.76(m,8H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((1-hydroxycycloheptyl) ethynyl) phenyl) propyl) acetamide followed by flash chromatography (9:1 CH) ₂Cl₂MeOH to 9:1 to 8:2CH₂Cl₂Containing 10% concentrated NH₄MeOH with OH) gave example 59. Dissolving the obtained oil (wetoil) containing water in CH₂C_l2In combination with MgSO₄Drying and concentration under reduced pressure gave example 59 as a clear oil which solidified to a white solid upon standing. Yield (0.41g, 36%):¹HNMR(400MHz,DMSO-d₆)7.32(brs,1H),7.24-7.29(m,2H),7.19-7.22(m,1H),5.27(s,1H),4.65(t,J=6.4Hz,1H),2.55-2.65(m,2H),1.94(dd,J=13.6,7.6Hz,2H),1.74-1.79(m,2H),1.42-1.66(m,10H)。

example 60

Preparation of N- (3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide

N- (3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide was prepared according to the procedure shown in equation 11.

Reaction formula 11

Example 11

1- ((3- (3-aminopropyl) phenyl) ethynyl) cyclohexanol (example 11) (0.057g,0.19mmol) was placed in acetic anhydride (2.0ml) and stirred at room temperature for 2 h. Water (10ml) was added under vortexing and sonication. The product was collected by filtration and washed with water (2X 5ml) and dried under vacuum overnight to give example 60 as a white solid. Yield (0.050g, 76%):¹HNMR(400MHz,DMSO-d₆)7.81(bs,1H),7.25(t,J=8.0Hz,1H),7.22(s,1H),7.16-7.19(m,2H),5.38(s,1H),2.99(q,J=5.6Hz,2H),2.54(t,J=7.2Hz,2H),1.80-1.84(m,2H),1.78(s,3H),1.43-1.68(m,9H),1.20-1.23(m,1H)。

example 61

Preparation of 3- (3- (pyridin-2-ylethynyl) phenyl) propan-1-amine

3- (3- (pyridin-2-ylethynyl) phenyl) propan-1-amine was prepared according to the procedure described in equation 12.

Reaction formula 12

Step 1: except thatCoupling of alcohol 11 with phthalimide following the procedure described in example 17, except using diisopropyl azodicarboxylate instead of diethyl azodicarboxylate, gave alkyne 52. Yield (6g, 76%): ¹HNMR(400MHz,CDCl₃)7.83(dd,J=5.2,2.8Hz,2H),7.70(dd,J=5.6,3.2Hz,2H),7.33(s,1H),7.24-7.29(m,1H),7.16-7.22(m,2H),3.74(t,J=7.2Hz,2H),3.04(s,1H),2.66(t,J=8.0Hz,2H),2.02(quint,J=7.2Hz,2H)。

Step 2: coupling of 2-bromopyridine with alkyne 52 as described in example 17 gave 2- (3- (3- (pyridin-2-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione (53) as a tan oil. Yield (0.6g, 80%):¹HNMR(400MHz,CDCl₃)：8.62(d,J=4.1Hz,1H),7.84(dd,J=5.6,3.2Hz,2H),7.71(dd,J=5.6,3.2Hz,2H),7.66-7.69(m,1H),7.52(d,J=8.0Hz,1H),7.44(s,1H),7.38(dt,J=7.2,1.6Hz,1H),7.20-7.28(m,3H),3.76(t,J=7.2Hz,2H),2.69(t,J=8.4Hz,2H),2.04(quint,J=7.6Hz,2H)。

and step 3: 2- (3- (3- (pyridin-2-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione (53) was deprotected according to the procedure described in example 17, except that the reaction was carried out at room temperature overnight. Purification by preparative HPLC (method 001P) gave example 61 trifluoroacetate as a tan oil. Suspending trifluoroacetate in CH₂Cl₂(15mL) and shaken with an aqueous ammonia solution (12.5%,20 mL). The organic layer was washed with water, followed by brine, and anhydrous Na₂SO₄Drying and concentration under reduced pressure gave example 61 as a tan oil. Yield (0.20g, 35%):¹HNMR(400MHz,DMSO-d₆)8.65(d,J=4.8Hz,1H),7.81(dt,J=7.6,1.6Hz,1H),7.60(d,J=7.6Hz,1H),7.24-7.44(m,4H),2.68(t,J=7.6Hz,2H),2.57(t,J=6.8Hz,2H),1.70(quint,J=6.8Hz,2H)。

example 62

Preparation of 2- (3- (pyridin-3-ylethynyl) phenoxy) ethylamine

2- (3- (pyridin-3-ylethynyl) phenoxy) ethylamine was prepared according to the method shown in reaction formula 13.

Reaction formula 13

Step 1: coupling of 3-bromophenol (14) with N-Boc-ethanolamine as used in example 17 gave bromide 54 as a pale yellow oil. Yield (8.34g, 91%):¹HNMR(400MHz,CDCl₃)7.14(t,J=8.0Hz,1H),7.09(dd,J=8.0,1.6Hz,1H),7.05(t,J=1.6Hz,1H),6.82(ddd,J=8.0,2.4,1.6Hz,1H),4.95(bs,1H),4.00(t,J=5.2Hz,2H),3.53(quint,J=5.2Hz,2H),1.45(s,9H)。

step 2: coupling of bromide 54 with alkyne 9 following the procedure used in example 17 gave alkyne 55 as a brown solid. Yield (0.90g, 90%): ¹HNMR(400MHz,CDCl₃),7.21(t,J=8.0Hz,1H),7.02(d,J=7.6Hz,1H),6.93-6.95(m,1H),6.85(ddd,J=8.4,2.8,0.8Hz,1H),4.97(brs,1H),4.01(t,J=5.2Hz,2H),3.51-3.52(m,2H),1.62,(s,6H),1.56(s,9H)。

And step 3: treatment of the alkyne 55 with KOH as used in example 17 gave alkyne 56 as a tan oil. Yield (0.2g, 80%):¹HNMR(400MHz,CDCl₃),7.23(d,J=8.0Hz,1H),7.10(dt,J=7.6,1.2Hz,1H),7.00-7.02(m,1H),6.90(ddd,J=8.4,2.8,0.8Hz,1H),4.97(brs,1H),4.01(t,J=5.2Hz,2H),3.49-3.54(m,2H),3.06(s,1H),1.45(s,9H)。

and 4, step 4: alkyne 56 is coupled with 3-bromopyridine as used in example 17 to give alkyne 57 as a tan oil. Yield (0.340g, 44%):¹HNMR(400MHz,CDCl₃),8.76(d,J=1.4Hz,1H),8.55(dd,J=4.8,1.2Hz,1H),7.81(dt,J=8.0,1.6Hz,1H),7.29(t,J=4.4Hz,1H),7.28(s,1H),7.16(d,J=8.0Hz,1H),7.06(brs,1H),6.92(dd,J=8.4,2.8Hz,1H),4.05(t,J=5.2Hz,2H),3.54(q,J=5.2Hz,2H),1.46(s,9H)。

and 5: deprotection of alkyne 57 with HCl/dioxane following the procedure used in example 36 gave example 62 hydrochloride as an off-white solid. Yield (0.230g,83%):¹HNMR(400MHz,DMSO-d₆)8.78(brs,1H),8.60(dd,J=4.8,1.6Hz,1H),8.11(brs,3H),8.02-8.04(m,1H),7.51(dd,J=8.0,5.2Hz,1H),7.37(t,J=8.0Hz,1H),7.20(d,J=8.0Hz,1H),7.18(d,J=1.6Hz,1H),7.06(dd,J=8.4,2.4Hz,1H),4.20(t,J=4.8Hz,2H),3.19(dd,J=10.4,5.6Hz,2H)。

example 63

Preparation of 3- (3- (pyridin-4-ylethynyl) phenyl) propan-1-amine

3- (3- (pyridin-4-ylethynyl) phenyl) propan-1-amine was prepared according to the method used in example 61.

Step 1: 4-bromopyridine was coupled with alkyne 52 and purified by flash chromatography (15% EtOAc-hexanes) to afford 2- (3- (3- (pyridin-4-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a yellow solid. Yield (0.271g, 53%):¹HNMR(400MHz,DMSO-d₆),8.58(dd,J=4.4,1.6Hz,2H),7.48(dd,J=4.4,1.6Hz,2H),7.41(s,1H),7.38(d,J=7.6Hz,1H),7.33(t,J=7.6Hz,1H),7.27(d,J=7.6Hz,1H),2.59(t,J=7.6Hz,2H),2.49(t,J=6.4Hz,2H),1.60(quint,J=7.2Hz,2H)。

step 2: deprotection of 2- (3- (3- (pyridin-4-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione affords example 63 as a yellow oil after conversion to the free base. Yield (0.023g, 13%):¹HNMR(400MHz,DMSO-d₆)8.58(d,J=4.0Hz,2H),7.85-7.52,(m,4H),7.48(d,J=5.2Hz,2H),7.44(s,1H),7.34(m,1H),7.30(d,J=5.2Hz,2H),3.57(t,J=6.9Hz,2H),2.63(t,J=7.4Hz,2H),1.89(quint,J=8.0Hz,2H)。

example 64

Preparation of 3- (3- (pyridin-3-ylethynyl) phenyl) propan-1-amine

3- (3- (pyridin-3-ylethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: coupling of 3-bromopyridine with alkyne 52 affords 2- (3- (3- (pyridin-4-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a tan oil. Yield (0.032g,42%):¹HNMR(400MHz,CDCl₃)　.　.8.76(d,J=1.6Hz,1H),8.55(dd,J=5.2,1.6Hz,1H),7.83(dd,J=5.2,2.8Hz,2H),7.80(dt,J=8.0,1.6Hz,1H),7.71(dd,J=5.2,2.8Hz,2H),7.39(s,1H),7.16-7.24(m,4H),3.76(t,J=7.2Hz,2H),2.70(t,J=8.0Hz,2H),2.05(quint,J=7.2Hz,2H)。

step 2: deprotection of 2- (3- (3- (pyridin-4-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione afforded example 64 as a yellow oil after conversion to the free base. Yield (0.135g, 65%):¹HNMR(400MHz,DMSO-d₆)8.71(d,J=2.0Hz,1H),8.55(dd,J=5.2,2.0Hz,1H),8.55(dt,J=8.0,1.6Hz,1H),7.43(dd,J=8.0,4.8Hz,1H),7.39(s,1H),7.38(d,J=7.6Hz,1H),7.31(t,J=7.2Hz,1H),7.25(d,J=7.6Hz,1H),3.30(obsm,2H),2.59(t,J=7.6Hz,2H),1.60(quint,J=8.4Hz,2H)。

example 65

3- (3- (thien-2-ylethynyl) phenyl) propan-1-amine

3- (3- (thien-2-ylethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: alkyne 52 was coupled with 2-bromothiophene and purified by flash chromatography (15% EtOAc-hexanes) to give 2- (3- (3- (thien-2-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a yellow solid. Yield (0.490g, 50%):¹HNMR(400MHz,CDCl₃)7.84(dd,J=5.6,3.2Hz,2H),7.71(dd,J=5.2,3.2Hz,2H),7.35(s,1H),7.26-7.30(m,3H),7.23(t,J=7.6Hz,1H),7.18(d,J=7.6Hz,1H),7.01(dd,J=5.2,3.6Hz,1H),3.76(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),2.05(quint.,J=7.6Hz,2H)。

step 2: deprotection of 2- (3- (3- (thien-2-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione follows the procedure of example 61. The reaction mixture was diluted with diethyl ether and the precipitate was removed by filtration. The filtrate was concentrated under reduced pressure and the diethyl ether precipitation step was repeated. Purification by preparative HPLC (method 001) afforded example 65 trifluoroacetate as a cream solid. Yield (0.210g, 65%): ¹HNMR(400MHz,CDCl₃)7.94(brs,3H),7.33(d,J=7.6Hz,1H),7.25-7.28(m,2H),7.22(d,J=7.6Hz,1H),7.09(d,J=7.6Hz,1H),6.99(dd,J=5.2,3.6Hz,1H),2.89(t,J=7.2Hz,2H),2.63(t,J=7.6Hz,2H),1.92-1.99(m,2H)。

Example 66

Preparation of 3- (3- (thien-3-ylethynyl) phenyl) propan-1-amine

3- (3- (thien-3-ylethynyl) phenyl) propan-1-amine was prepared according to the method described in example 61.

Step 1: alkyne 52 was coupled with 3-bromothiophene and purified by flash chromatography (17% EtOAc-hexanes) to give 2- (3- (3- (thien-3-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione as an off-white solid. Yield (0.441g, 43%):¹HNMR(400MHz,CDCl₃)7.84(dd,J=5.2,3.2Hz,2H),7.71(dd,J=5.2,3.2Hz,2H),7.51(dd,J=2.8,1.2Hz,1H),7.36(s,1H),7.23(t,J=7.6Hz,1H),7.19(dd,J=4.8,1.2Hz,1H),7.17(d,J=7.6Hz,1H),3.76(t,J=6.8Hz,2H),2.68(t,J=7.6Hz,2H),2.04(quint.,J=7.6Hz,2H)。

step 2: deprotection of 2- (3- (3- (thiophen-3-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione according to the procedure used in example 65 gave example 66 as a tan oil, except that no HPLC purification step was required. Yield (0.190g,66%):¹HNMR(400MHz,CDCl₃)7.51-7.52(m,1H),7.15-7.36(m,6H),2.74(t,J=7.2Hz,2H),2.66(t,J=7.9Hz,2H),1.75-1.83(m,2H),1.54(brs,2H)。

example 67

Preparation of 3- (3- (6-methoxyhex-1-ynyl) phenyl) prop-1-amine

3- (3- (6-Methoxyhex-1-ynyl) phenyl) prop-1-amine was prepared according to the method described for example 36.

Step 1: aryl bromide 34 was coupled with 6-methoxyhex-1-yne following the procedure used for the preparation of example 36 and flash chromatography (10% EtOAc-hexanes) afforded tert-butyl 3- (3- (6-methoxyhex-1-ynyl) phenyl) propylcarbamate as a tan oil. Yield (0.20g, 36%).

Step 2: except for CH₂Cl₂Used as co-solvent for the reaction (HCl-dioxane solution: CH) ₂Cl₂7:5) tert-butyl 3- (3- (6-methoxyhex-1-ynyl) phenyl) propylcarbamate was deprotected according to the method used in example 36 and purified by preparative HPLC (method 004P) to give example 67 hydrochloride as an off-white solid. Yield (0.050g, 30%):¹HNMR(400MHz,CDCl₃)8.37(brs,3H),7.10-7.24(m,4H),4.02(t,J=6.4Hz,2H),3.78(s,3H),2.98(t,J=7.6Hz,2H),2.69(t,J=7.6Hz,2H),2.44(t,J=7.6Hz,2H),2.04-2.12(m,2H),1.82-1.89(m,2H),1.63-1.73(m,2H)。

example 68

Preparation of 6- (3- (3-aminopropyl) phenyl) hex-5-yn-1-ol

6- (3- (3-aminopropyl) phenyl) hex-5-yn-1-ol was prepared according to the method used in example 36.

Step 1: hex-5-yn-1-ol was coupled with bromide 34 as used in example 36. Purification by flash chromatography (30% EtOAc-hexanes) afforded tert-butyl 3- (3- (6-hydroxyhex-1-ynyl) phenyl) propylcarbamate as a white solid. Yield (0.350g,66%):¹HNMR(400MHz,CDCl₃)7.17-7.23(m,3H),7.07-7.10(m,1H),6.81-6.84(m,1H),4.53(brs,1H),3.72(q,J=6.0Hz,2H),3.10-3.18(m,2H),2.60(t,J=8.0Hz,2H),2.46(t,J=6.8Hz,2H),1.63-1.83(m,6H),1.44(s,9H)。

step 2: deprotection of tert-butyl 3- (3- (6-hydroxyhex-1-ynyl) phenyl) propylcarbamate followed by purification by preparative HPLC using method 001P gave example 68 as a white solid. Yield (0.140g, 34%):¹HNMR(400MHz,CDCl₃)7.21(d,J=7.6Hz,2H),7.17(t,J=7.6Hz,1H),7.07(dm,J=7.2Hz,1H),3.68(t,J=6.4Hz,2H),2.95(t,J=7.6Hz,2H),2.67(t,J=7.6,2H),2.43(t,J=6.4Hz,2H),2.06(quint.,J=7.6Hz,2H),1.71-1.79(m,2H),1.61-1.68(m,2H)。

example 69

Preparation of 3-amino-1- (3- (4-phenylbutan-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (4-phenylbut-1-ynyl) phenyl) propan-1-ol was prepared according to the general reaction scheme used in modified example 19.

Step 1: aryl bromide 25 was coupled with but-3-ynylbenzene as used in example 1 and 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-phenylbut-1-ynyl) phenyl) propyl) acetamide was obtained as a tan oil by flash chromatography (20% EtOAc-hexanes). Yield (0.340g, 52%): ¹HNMR(400MHz,CDCl₃)7.28-7.36(m,7H),7.24-7.27(m,2H),4.84-4.88(m,1H),3.66-3.74(m,1H),3.41(ddd,J=17.6,8.0,4.4Hz,1H),2.93(t,J=7.6Hz,2H),2.70(t,J=7.6Hz,2H),2.27(d,J=1.6Hz,1H),1.90-2.03(m,2H)。

Step 2: 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-phenylbut-1-ynyl) phenyl) propyl) acetamide was deprotected as used in example 1 except that the reaction was heated overnight. Purification by preparative HPLC (method 004P) gave example 69 as a brown solid. Yield (0.085g, 33%):¹HNMR(400MHz,DMSO-d₆)7.21-7.34(m,9H),4.67(t,J=6.0Hz,1H),2.88(t,J=7.2Hz,2H),2.68-2.74(m,4H),1.68(q,J=6.4Hz,2H),0.86-0.92(m,1H)。

example 70

Preparation of 3- (3- (5-methoxypent-1-ynyl) phenyl) propan-1-amine

3- (3- (5-Methoxypent-1-ynyl) phenyl) propan-1-amine was prepared according to the procedure used in example 36.

Step 1: aryl bromide 34 was coupled with 5-methoxypent-1-yne and purified by flash chromatography (10% ethyl acetate-hexanes) to give a yellow oilTert-butyl 3- (3- (5-methoxypent-1-ynyl) phenyl) propylcarbamate in the form of a solid. Yield (0.170g, 32%):¹HNMR(400MHz,CDCl₃)7.17-7.23(m,3H),7.08-7.10(m,1H),4.52(brs,1H),4.31(t,J=6.4Hz,2H),3.79(s,3H),3.08-3.17(m,2H),2.60(t,J=7.6Hz,2H),2.53(t,J=7.2Hz,2H),1.94-2.01(m,2H),1.76-1.83(m,2H),1.44(s,9H)。

step 2: deprotection of tert-butyl 3- (3- (5-methoxypent-1-ynyl) phenyl) propylcarbamate and purification by preparative HPLC (method 001P) afforded example 70 trifluoroacetate as a white solid. Yield (0.110g, 92%):¹HNMR(400MHz,CDCl₃)7.11-7.24(m,4H),4.30(t,J=6.4Hz,2H),3.78(s,3H),2.97(t,J=7.2Hz,2H),2.70(t,J=7.6Hz,2H),2.52(t,J=7.2Hz,2H),2.02-2.11(m,2H),1.94-2.00(m,2H)。

example 71

Preparation of 3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol was prepared according to the method used in example 69.

Example 72

Preparation of 3- (3- (4-phenylbutan-1-ynyl) phenyl) propan-1-amine

3- (3- (4-phenylbutan-1-ynyl) phenyl) propan-1-amine was prepared according to the method used in example 36.

Step 1: aryl bromide 34 was coupled with but-3-ynylbenzene as used in example 36. Purification by flash chromatography (10% ethyl acetate-hexanes) afforded tert-butyl 3- (3- (4-phenylbut-1-ynyl) phenyl) propylcarbamate as a brown oil. Yield (0.40g, 82%).

Step 2: deprotection of tert-butyl 3- (3- (4-phenylbut-1-ynyl) phenyl) propylcarbamate afforded example 72 hydrochloride as a white solid.¹HNMR(400MHz,DMSO-d₆)7.74(brs,3H),7.14-7.28(m,9H),2.82(t,J=7.2Hz,2H),2.71(t,J=7.6Hz,2H),2.67(t,J=7.2Hz,2H),2.58(t,J=7.6Hz,2H),1.78(quint.,J=7.6Hz,2H)。

Example 73

Preparation of 2- (3- (4-methylpent-1-ynyl) phenoxy) ethylamine

2- (3- (4-methylpent-1-ynyl) phenoxy) ethylamine was prepared as described in equation 14.

Reaction formula 14

Step 1: aryl bromide 54 was coupled with 4-methylpent-1-yne and purified by flash chromatography (20% ethyl acetate-hexanes) as used in example 1 to give alkyne 58 as a brown oil. Yield (0.289g, 48%).

Step 2: alkyne 58 was deprotected and purified by preparative HPLC (method 004P) as used in example 36 to give the hydrochloride salt of example 73 as a white solid. Yield (0.040g, 20%):¹HNMR(400MHz,DMSO-d₆)8.11(brs,3H),7.25(t,J=8.4Hz,1H),6.92-6.98(m,3H),4.14(t,J=4.8Hz,2H),3.15(t,J=4.8Hz,2H),2.28(d,J=6.4Hz,2H),1.78-1.84(m,1H),0.96(d,J=6.8Hz,6H)。

example 74

Preparation of 6- (3- (2-aminoethoxy) phenyl) hex-5-yn-1-ol

6- (3- (2-Aminoethoxy) phenyl) hex-5-yn-1-ol was prepared according to the method used in example 73.

Step 1: aryl bromide 54 was coupled with hex-5-yn-1-ol and purified by flash chromatography (15% ethyl acetate-hexanes) to give tert-butyl 2- (3- (6-hydroxyhex-1-ynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.500g, 79%):¹HNMR(400MHz,CDCl₃)7.17(t,J=7.6Hz,1H),6.99(d,J=7.6Hz,1H),6.91(s,1H),6.81(d,J=8.4Hz,1H),4.72(brs,1H),4.00(t,J=4.8Hz,2H),3.72(m,2H),3.48-3.55(m,2H),2.46(d,J=6.8Hz,2H),1.66-1.79(m,4H),1.58(s,1H),1.45(s,9H)。

step 2: deprotection of tert-butyl 2- (3- (6-hydroxyhex-1-ynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 74 hydrochloride as a brown solid. Yield (0.161g, 40%):¹HNMR(400MHz,DMSO-d₆)8.15(brs,3H),7.22-7.27(m,1H),6.92-6.97(m,3H),4.14(t,J=5.0Hz,2H),3.40(t,J=5.8Hz,2H),3.14-3.15(m,2H),2.39(t,J=6.6Hz,2H),1.51-1.53(m,4H)。

example 75

Preparation of 2- (3- (3-phenylprop-1-ynyl) phenoxy) ethylamine

2- (3- (3-phenylprop-1-ynyl) phenoxy) ethylamine was prepared by the method used in example 73.

Step 1: aryl bromide 54 was coupled with prop-2-ynylbenzene and purified by flash chromatography (5% ethyl acetate-hexanes) to give tert-butyl 2- (3- (3-phenylprop-1-ynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.530g, 79%):¹HNMR(400MHz,CDCl₃)7.33-7.42(m,3H),7.05-7.23(m,4H),6.97(s,1H),6.81-6.85(m,1H),4.97(brs,1H),4.01(t,J=4.8Hz,2H),3.83(s,2H),3.52(q,J=4.8Hz,2H),1.45(s,9H)。

step 2: deprotection of tert-butyl 2- (3- (3-phenylprop-1-ynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 75 hydrochloride as a cream solid. Yield (0.208g, 54%): ¹HNMR(400MHz,DMSO-d₆)8.29(brs,2H),7.20-7.38(m,6H),6.95-7.04(m,3H),4.17(t,J=5.2Hz,2H),3.85(s,3H),3.14(t,J=5.2Hz,2H)。

Example 76

Preparation of 4- (3- (2-aminoethoxy) phenyl) but-3-yn-1-ol

4- (3- (2-Aminoethoxy) phenyl) but-3-yn-1-ol was prepared according to the method used in example 73.

Step 1: aryl bromide 54 was coupled with but-3-yn-1-ol and purified by flash chromatography (35% ethyl acetate-hexanes) to give tert-butyl 2- (3- (4-hydroxybut-1-ynyl) phenoxy) ethylcarbamate as a brown oil contaminated with the alkyne dimer. Yield (0.296g, 51%):¹HNMR(400MHz,CDCl₃)7.20(t,J=8.0Hz,1H),7.02(d,J=7.6Hz,1H),6.94(s,1H),6.84(ddd,J=8.4,2.8,0.8Hz,1H),4.98(brs,1H),4.00(t,J=5.2Hz,2H),3.83(q,J=6.4Hz,2H),3.53(q,J=5.2Hz,2H),2.69(t,J=6.4Hz,2H),1.83(t,J=6.0Hz,1H),1.45(s,9H)。

step 2: deprotection of tert-butyl 2- (3- (4-hydroxybut-1-ynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 76 hydrochloride as an off-white solid. Yield (0.064g, 32%):¹HNMR(400MHz,DMSO-d₆)8.07(brs,3H),7.25(t,J=8.0Hz,1H),6.92-6.99(m,3H),4.87-4.90(m,1H),4.14(t,J=5.2Hz,2H),3.54(q,J=6.4Hz,2H),3.16(s,2H),2.51(t,J=6.4Hz,2H)。

example 77

Preparation of 2- (3- (hept-1-ynyl) phenoxy) ethylamine

2- (3- (hept-1-ynyl) phenoxy) ethylamine was prepared by the method used in example 73.

Step 1: aryl bromide 54 was coupled with 1-heptyne and purified by flash chromatography (15% ethyl acetate-hexanes) to give tert-butyl 2- (3- (hept-1-ynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.238g, 37%).

Step 2: deprotection of tert-butyl 2- (3- (hept-1-ynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 77 hydrochloride as a white solid. Yield (0.018g, 11%): ¹HNMR(400MHz,DMSO-d₆)7.95(brs,3H),7.25(t,J=8.4Hz,1H),6.92-6.97(m,3H),4.13(t,J=4.8Hz,2H),3.17(t,J=4.8Hz,2H),2.37(t,J=7.2Hz,2H),1.47-1.54(m,2H),1.24-1.39(m,4H),0.85(t,J=6.8Hz,3H)。

Example 78

Preparation of 1- ((3- (2-aminoethoxy) phenyl) ethynyl) -cycloheptanol

1- ((3- (2-Aminoethoxy) phenyl) ethynyl) -cycloheptanol was prepared according to the procedure used in example 18.

Step 1: the coupling of bromide 19 with 1-ethynylcycloheptanol was carried out as described for example 18, except that the reaction was heated for 2 hours. After cooling the reaction mixture to room temperature, it was diluted with ethyl acetate and washed with water. The combined organics were filtered through celite. Adding Na into the filtrate₂SO₄Dried and treated with activated carbon. After filtration, the solution was concentrated under reduced pressure. Purification by flash chromatography (10-50% etoac-hexanes gradient) afforded 2,2, 2-trifluoro-N- (2- (3- ((1-hydroxycycloheptyl) ethynyl) phenoxy) ethyl) acetamide as an orange oil. Yield (1.078g, 60%):¹HNMR(400MHz,CDCl₃)7.20(brs,1H),7.18(t,J=8.0Hz,1H),7.02(dt,J=7.2,0.8Hz,1H),6.90(dd,J=2.4,1.6Hz,1H),6.81(ddd,J=8.4,2.4,0.8Hz,1H),4.05(t,J=5.2Hz,2H),3.72(q,J=5.3Hz,2H),2.43(brs,1H),2.05-2.11(m,2H),1.84-1.91(m,2H),1.53-1.70(m,8H)。

step 2: to a solution of 2,2, 2-trifluoro-N- (2- (3- ((1-hydroxycycloheptyl) ethynyl) phenoxy) ethyl) acetamide (1.07g,2.9mmol) in methanol (20mL) was added saturated K₂CO₃Aqueous solution (. about.10 mL). The reaction mixture was stirred vigorously and heated at 50 ℃ for 2 hours. After removing volatile components by concentration under reduced pressure, the mixture was separated in ethyl acetate and water. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. Purification by flash chromatography (10% 7M ammonia in methanol-ethyl acetate) afforded example 78 as a light yellow solid. (yield 0.70g, 88%): ¹HNMR(400MHz,CDCl₃)7.19(t,J=8.0Hz,1H),7.01(dt,J=8.0,0.8Hz,1H),6.95(dd,J=2.8,1.6Hz,1H),6.85(ddd,J=8.4,2.4,1.2Hz,1H),3.97(t,J=4.8Hz,2H),3.07(brs,2H),2.08-2.13(m,2H),1.87-1.94(m,2H),1.59-1.74(m,11H)。

Example 79

Preparation of 2- (3- (cyclopentylethynyl) phenoxy) ethylamine

2- (3- (cyclopentylethynyl) phenoxy) ethylamine was prepared according to the method described for example 73.

Step 1: aryl bromide 54 was coupled with ethynylcyclopentane and purified by flash chromatography (20% ethyl acetate-hexanes) to give tert-butyl 2- (3- (cyclopentylethynyl) phenoxy) ethylcarbamate as a yellow oil. Yield (0.290g, 46%).

Step 2: deprotection of tert-butyl 2- (3- (cyclopentylethynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 79 hydrochloride as a white solid. Yield (0.100g, 49%):¹HNMR(400MHz,DMSO-d₆)8.24(brs,3H),7.23(dd,J=9.2,7.6Hz,1H),6.91-6.95(m,3H),4.15(t,J=5.2Hz,2H),3.13(t,J=5.2Hz,2H),2.77-2.85(m,1H),1.89-1.97(m,2H),1.52-1.71(m,6H)。

example 80

Preparation of 2- (3- (cyclohexylethynyl) phenoxy) ethylamine

2- (3- (cyclohexylethynyl) phenoxy) ethylamine was prepared according to the procedure described for example 73.

Step 1: aryl bromide 54 was coupled with ethynylcyclohexane and purified by flash chromatography (5-10% ethyl acetate-hexanes) to give tert-butyl 2- (3- (cyclohexylethynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.170g, 26%).

Step 2: deprotection of tert-butyl 2- (3- (cyclohexylethynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 80 hydrochloride as a white solid. Yield (0.023g, 16%): ¹HNMR(400MHz,DMSO-d₆)7.93(brs,3H),7.23-7.27(m,1H),6.92-6.97(m,3H),4.13(t,J=4.8Hz,2H),3.16(t,J=5.2Hz,2H),1.74-1.82(m,2H),1.60-1.68(m,2H),1.36-1.50(m,3H),1.28-1.48(m,4H)。

Example 81

Preparation of 2- (3- (phenylethynyl) phenoxy) ethylamine

2- (3- (phenylethynyl) phenoxy) ethylamine was prepared according to the method described for example 73.

Step 1: aryl bromide 54 was coupled with ethynylbenzene and purified by flash chromatography (22% ethyl acetate-hexanes) to give tert-butyl 2- (3- (phenylethynyl) phenoxy) ethylcarbamate as a yellow oil. Yield (0.200g, 31%).

Step 2: deprotection of tert-butyl 2- (3- (phenylethynyl) phenoxy) ethylcarbamate and purification as described in example 73 gave example 81 hydrochloride as a white solid. Yield (0.150g, 71%):¹HNMR(400MHz,DMSO-d₆)8.12(brs,3H),7.52-7.54(m,2H),7.40-7.41(m,3H),7.34(t,J=8.0Hz,1H),7.12-7.16(m,2H),7.02(dd,J=8.4,2.0Hz,1H),4.19(t,J=4.8Hz,2H),3.18(t,J=4.8Hz,2H)。

example 82

Preparation of 3- (3- (naphthalen-1-ylethynyl) phenyl) propan-1-amine

3- (3- (naphthalen-1-ylethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: alkyne 52 is coupled with 2-bromonaphthalene and purified by flash chromatography (3-5% ethyl acetate-hexane) to give 2- (3- (3- (naphthalen-1-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a brown oil. Yield (0.300g, 41%):¹HNMR(400MHz,CDCl₃)8.44(d,J=8.0Hz,1H),7.82-7.88(m,4H),7.76(dd,J=7.2,1.2Hz,1H),7.70(dd,J=5.2,3.2Hz,2H),7.59-7.64(m,1H),7.52-7.56(m,1H),7.42-7.49(m,3H),7.29(d,J=7.6Hz,1H),7.22(d,J=8.0Hz,1H),3.79(t,J=7.2Hz,2H),2.73(t,J=7.2Hz,2H),2.09(quint.,J=7.2Hz,2H)。

step 2: deprotection of 2- (3- (3- (naphthalen-1-ylethynyl) phenyl) propyl) isoindoline-1, 3-dione and purification as described for example 61 gave example 82 as a semi-solid. Yield (0.040g, 25%): ¹HNMR(400MHz,DMSO-d₆)8.18(s,1H),7.80-7.95(m,3H),7.69(brs,2H),7.57-7.62(m,3H),7.45-7.49(m,2H),7.40(t,J=7.6Hz,1H),7.30(d,J=7.6Hz,1H),2.80(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),1.87(quint,J=7.6Hz,2H)。

Example 83

Preparation of 3- (3- (o-tolylethynyl) phenyl) propan-1-amine

3- (3- (o-tolylethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: alkyne 52 was coupled with 1-ethynyl-2-methylbenzene and purified by flash chromatography (15% ethyl acetate-hexanes) to give 2- (3- (3- (o-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a brown oil. Yield (0.480g, 61%):¹HNMR(400MHz,CDCl₃)7.83(dd,J=5.6,3.2Hz,2H),7.76(dd,J=5.6,3.2Hz,2H),7.48(d,J=7.2Hz,1H),7.37(m,1H),7.31-7.32(m,1H),7.15-7.24(m,5H),3.77(t,J=7.2Hz,2H),2.70(t,J=7.2Hz,2H),2.50(s,3H),2.02-2.09(m,2H)。

step 2: deprotection of 2- (3- (3- (o-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione and purification by preparative HPLC (method 004P) afforded example 83 as a brown solid. Yield (0.080g, 25%):¹HNMR(400MHz,DMSO-d₆)7.24-7.53(m,8H),2.65(t,J=8.0Hz,2H),2.57(t,J=6.8Hz,2H),2.49(s,3H),1.65-1.72(m,2H)。

example 84

Preparation of 3- (3- (p-tolylethynyl) phenyl) propan-1-amine

3- (3- (p-tolylethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: alkyne 52 was coupled with 1-ethynyl-4-methylbenzene and purified by flash chromatography (10% ethyl acetate-hexanes) to give 2- (3- (3- (p-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a brown oil. Yield (0.487g, 61%):¹HNMR(400MHz,CDCl₃)7.83(dd,J=5.2,2.8Hz,2H),7.70(dd,J=5.2,2.8Hz,2H),7.41(d,J=8.0Hz,1H),7.14-7.36(m,7H),3.76(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),2.37(s,3H),2.01-2.09(m,2H)。

step 2: deprotection of 2- (3- (3- (p-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione and purification as described for example 61 afforded example 84 trifluoroacetate as a white solid. Yield (0.060g, 18%): ¹HNMR(400MHz,DMSO-d₆)7.53(brs,3H),7.31-7.41(m,5H),7.20-7.24(m,3H),2.74(t,J=7.6Hz,1H),2.63(t,J=7.2Hz,2H),2.31(s,3H),1.77-1.85(m,2H)。

Example 85

Preparation of 2- (3- (3-cyclopentylprop-1-ynyl) phenoxy) ethylamine

2- (3- (3-Cyclopentylprop-1-ynyl) phenoxy) ethylamine was prepared as described in example 73.

Step 1: prop-2-ynylcyclopentane was coupled with bromide 54 and purified by flash chromatography (15% ethyl acetate-hexanes) to give tert-butyl 2- (3- (3-cyclopentylprop-1-ynyl) phenoxy) ethylcarbamate as a white solid. Yield (0.500g, 76%):¹HNMR(400MHz,CDCl₃)6.91-7.21(m,3H),6.80-6.84(m,1H),4.97(brs,1H),4.00(t,J=4.8Hz,2H),3.52(q,J=4.4Hz,2H),2.40(d,J=6.8Hz,2H),2.07-2.17(m,1H),1.80-1.87(m,2H),1.48-1.70(m,4H),1.45(s,9H),1.29-1.40(m,2H)。

step 2: deprotection of tert-butyl 2- (3- (3-cyclopentylprop-1-ynyl) phenoxy) ethylcarbamate and purification by flash chromatography (7% methanol in dichloromethane) afforded example 85 hydrochloride as a white solid. Yield (0.160g, 45%):¹HNMR(400MHz,DMSO-d₆)7.20-7.24(m,1H),6.89-6.94(m,3H),4.00(t,J=5.0Hz,2H),3.00(brs,2H),2.04(quint.,J=7.2,2H),1.71-1.78(m,2H),1.44-1.62(m,4H),1.20-1.31(m,3H)。

example 86

Preparation of 2- (3- (4-phenylbut-1-ynyl) phenoxy) ethylamine

2- (3- (4-phenylbut-1-ynyl) phenoxy) ethylamine was prepared as described in example 73.

Step 1: but-3-ynylbenzene was coupled with bromide 54 and purified by flash chromatography (5-10% ethyl acetate-hexanes) to give tert-butyl 2- (3- (4-phenylbut-1-ynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.503g, 72%).

Step 2: deprotection of tert-butyl 2- (3- (4-phenylbut-1-ynyl) phenoxy) ethylcarbamate and purification by flash chromatography (1-10% methanol in dichloromethane with minor amounts of triethylamine) afforded example 86 as a yellow solid. Yield (0.127g, 33%): ¹HNMR(400MHz,CDCl₃)7.27-7.28(m,4H),7.17-7.22(m,2H),6.82-6.89(m,3H),3.88(t,J=5.6Hz,2H),2.80-2.86(m,4H),2.66(t,J=7.2Hz,2H)。

Example 87

Preparation of 3- (3- (m-tolylethynyl) phenyl) propan-1-amine

3- (3- (m-tolylethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: alkyne 52 was coupled with 3-iodotoluene and purified by flash chromatography (10% ethyl acetate-hexanes) to give 2- (3- (3- (m-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione as a brown oil. Yield (0.396g, 50%):¹HNMR(400MHz,CDCl₃)8.40(dd,J=5.6,3.2Hz,2H),7.71(dd,J=5.2,3.2Hz,2H),7.13-7.36(m,8H),3.76(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),2.36(s,3H),2.01-2.09(m,2H)。

step 2: deprotection of 2- (3- (3- (m-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione and purification as described for example 61 afforded example as an off-white solid87 trifluoroacetic acid ester. Yield (0.030g, 12%):¹HNMR(400MHz,DMSO-d₆)7.60(brs,3H),7.21-7.39(m,8H),2.75(t,J=8.0Hz,1H),2.63(t,J=7.6Hz,2H),2.29(s,3H),1.77-1.85(m,2H)。

example 88

Preparation of 3- ((3- (2-aminoethoxy) phenyl) ethynyl) benzonitrile

3- ((3- (2-aminoethoxy) phenyl) ethynyl) benzonitrile is prepared according to the method described for example 62.

Step 1: alkyne 56 was coupled with 3-bromobenzylnitrile and purified by flash chromatography (15% ethyl acetate-hexanes) to give tert-butyl 2- (3- ((3-cyanophenyl) ethynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.275g, 39%):¹HNMR(400MHz,CDCl₃)7.81(s,1H),7.73(dt,J=8.0,1.2Hz,1H),7.61(dt,J=7.6,1.2Hz,1H),7.47(t,J=8.0Hz,1H),7.28-7.31(m,1H),7.15(d,J=7.6Hz,1H),7.05(s,1H),6.93(dd,J=8.0,2.0Hz,1H),4.98(brs,1H),4.05(t,J=5.2Hz,2H),3.55(q,J=5.2Hz,2H),1.46(s,9H)。

step 2: deprotection of tert-butyl 2- (3- ((3-cyanophenyl) ethynyl) phenoxy) ethylcarbamate afforded example 88 hydrochloride as a white solid. Yield (0.195g, 97%): ¹HNMR(400MHz,DMSO-d₆)8.03(s,4H),7.85-7.88(m,2H),7.62(t,J=8.0Hz,1H),7.37(t,J=8.0Hz,1H),7.16-7.20(m,2H),7.06(dd,J=8.4,2.4Hz,1H),4.19(t,J=4.8Hz,2H),3.20(q,J=4.8Hz,2H)。

Example 89

Preparation of 2- ((3- (3-aminopropyl) phenyl) ethynyl) phenol

2- ((3- (3-aminopropyl) phenyl) ethynyl) phenol was prepared according to the procedure described for example 61.

Step 1: alkyne 52 was coupled with 2-iodophenol and purified by flash chromatography (8% ethyl acetate-hexanes) to give 2- (3- (3- ((2-hydroxyphenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione as a yellow oil. Yield (0.550g, 69%).

Step 2: deprotection of 2- (3- (3- (m-tolylethynyl) phenyl) propyl) isoindoline-1, 3-dione and purification as described for example 61 afforded example 89 trifluoroacetate as an off white solid. Yield (0.195g, 53%):¹HNMR(400MHz,CD₃OD)7.75(s,1H),7.69-7.71(m,1H),7.58(d,J=7.6Hz,1H),7.48-7.51(m,1H),7.33-7.38(m,1H),7.19-7.29(m,3H),7.14-7.15(m,1H),2.67-2.74(m,4H),1.80-1.88(m,2H)。

example 90

Preparation of 3- ((3- (3-aminopropyl) phenyl) ethynyl) benzonitrile

3- ((3- (3-aminopropyl) phenyl) ethynyl) benzonitrile was prepared according to the method described for example 61.

Step 1: alkyne 52 was coupled with 3-bromobenzylnitrile and purified by flash chromatography (20% ethyl acetate-hexanes) to give 3- ((3- (3- (1, 3-dioxoisoindolin-2-yl) propyl) phenyl) ethynyl) benzonitrile as a yellow oil. Yield (0.456g, 56%):¹HNMR(400MHz,CDCl₃)7.84(dd,J=5.6,2.8Hz,2H),7.80(t,J=1.2Hz,1H),7.70-7.74(m,3H),7.60(dt,J=7.6,1.2Hz,1H),7.47(t,J=8.0Hz,1H),7.39(s,1H),7.21-7.33(m,3H),3.76(t,J=7.2Hz,2H),2.70(t,J=7.6Hz,2H),2.02-2.09(m,2H)。

step 2: 3- ((3- (3- (1, 3-dioxoisoindolin-2-yl) propyl) phenyl) ethynyl) benzonitrile was deprotected and purified as described for example 61 and converted to the free base to give example 90 as a yellow oil. Yield (0.085g, 28%): ¹HNMR(400MHz,DMSO-d₆)8.02(t,J=1.2Hz,1H),7.85(dd,J=8.0,1.6Hz,2H),7.60(t,J=8.0Hz,1H),7.39(s,1H),7.36(dt,J=7.6,1.2Hz,1H),7.32(t,J=7.6Hz,1H),7.26(dt,J=7.2,1.2Hz,1H),2.59(t,J=7.6Hz,2H),2.49(t,J=7.2Hz,2H),1.57-1.64(m,2H)。

Example 91

Preparation of 2- (3- ((2-methoxyphenyl) ethynyl) phenoxy) ethylamine

2- (3- ((2-methoxyphenyl) ethynyl) phenoxy) ethylamine was prepared according to the method described for example 73.

Step 1: 1-ethynyl-2-methoxybenzene was coupled with bromide 54 and purified by flash chromatography (12% ethyl acetate-hexanes) to give tert-butyl 2- (3- ((2-methoxyphenyl) ethynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.500g, 44%):¹HNMR(400MHz,CDCl₃)7.49(dd,J=7.6,1.6Hz,1H),7.23-7.34(m,2H),7.18(dt,J=7.6,1.2Hz,1H),7.06-7.08(m,1H),6.86-6.96(m,3H),4.99(brs,1H),4.04(t,J=5.2Hz,2H),3.97(s,3H),3.54(q,J=5.2Hz,2H),1.46(s,9H)。

step 2: deprotection of tert-butyl 2- (3- ((2-methoxyphenyl) ethynyl) phenoxy) ethylcarbamate and purification by flash chromatography (6% methanol in dichloromethane) afforded example 91 hydrochloride as a white solid. Yield (0.160g, 43%):¹HNMR(400MHz,DMSO-d₆)7.44(dd,J=7.6,1.6Hz,1H),7.30-7.39(m,2H),7.06-7.11(m,3H),6.93-7.01(m,2H),4.13(t,J=5.2Hz,2H),3.82(s,3H),3.12(t,J=5.2Hz,2H)。

example 92

Preparation of 3- (3- ((3- (trifluoromethyl) phenyl) ethynyl) phenyl) propan-1-amine

3- (3- ((3- (trifluoromethyl) phenyl) ethynyl) phenyl) propan-1-amine was prepared according to the procedure described for example 61.

Step 1: alkyne 52 was coupled with 3-bromobenzotriflate (3-bromobenzotriflate) and purified by flash chromatography (5% ethyl acetate-hexanes) to give 2- (3- (3- ((3- (trifluoromethyl) phenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione as a brown oil. Yield (0.477g, 53%): ¹HNMR(400MHz,CDCl₃)7.84(dd,J=5.2,3.2Hz,2H),7.79(s,1H),7.68-7.72(m,3H),7.57(d,J=8.0Hz,1H),7.48(t,J=7.6Hz,1H),7.39(s,1H),7.32(dt,J=7.2,1.2Hz,1H),7.26(t,J=7.6Hz,1H),7.21(d,J=7.6Hz,1H),3.77(t,J=7.2Hz,2H),2.70(t,J=7.2Hz,2H),2.02-2.09(m,2H)。

Step 2: deprotection of 2- (3- (3- ((3- (trifluoromethyl) phenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione, and purification and conversion to the free base as described for example 61 gave example 92 as a semi-solid. Yield (0.240g, 71%):¹HNMR(400MHz,DMSO-d₆)7.88(s,1H),7.82(d,J=8.0Hz,1H),7.74(d,J=8.0Hz,1H),7.64(t,J=7.6Hz,1H),7.41(s,1H),7.37(d,J=7.6Hz,1H),7.32(t,J=7.6Hz,1H),7.25(d,J=7.2Hz,1H),2.59(t,J=7.6Hz,2H),2.50(t,J=6.8Hz,2H),1.58-1.65(m,2H)。

example 93

Preparation of 3- (3- ((3, 5-di-tert-butylphenyl) ethynyl) phenyl) propan-1-amine

3- (3- ((3, 5-di-tert-butylphenyl) ethynyl) phenyl) propan-1-amine was prepared according to the method described for example 61.

Step 1: alkyne 52 was coupled with 1-bromo-3, 5-di-tert-butylbenzene and purified by flash chromatography (12% ethyl acetate-hexanes) to give 2- (3- (3- ((3, 5-di-tert-butylphenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione as a colorless oil. Yield (0.410g, 41%):¹HNMR(400MHz,CDCl₃)7.84(dd,J=5.2,3.2Hz,2H),7.71(dd,J=5.6,3.2Hz,2H),7.15-7.40(m,7H),3.77(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),2.01-2.09(m,2H),1.34(s,18H)。

step 2: 2- (3- (3- ((3, 5-di-tert-butylphenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione was deprotected and purified as described for example 61 and converted to the free base to give example 93 as a colorless oil. Yield (0.150g, 50%):¹HNMR(400MHz,DMSO-d₆)7.41(t,J=1.6Hz,1H),7.37(s,1H),7.32-7.34(m,3H),7.29(t,J=7.6Hz,1H),7.20(d,J=7.2Hz,1H),2.58(t,J=7.6Hz,2H),2.49(t,J=6.8Hz,2H),1.57-1.65(m,2H),1.27(s,18H)。

example 94

Preparation of 3- (3- ((4- (methylthio) phenyl) ethynyl) phenyl) propan-1-amine

3- (3- ((4- (methylthio) phenyl) ethynyl) phenyl) propan-1-amine was prepared according to the procedure described for example 61.

Step 1: alkyne 52 was coupled with 4-bromothioanisole and purified by flash chromatography (16% ethyl acetate-hexanes) to give 2- (3- (3- ((4- (methylthio) phenyl) ethane) as a brown oilAlkynyl) phenyl) propyl) isoindoline-1, 3-dione. Yield (0.160g, 32%):¹HNMR(400MHz,CDCl₃)7.83(dd,J=5.6,3.2Hz,2H),7.71(dd,J=5.2,2.8Hz,2H),7.43(dt,J=8.8,2.0Hz,2H),7.36(s,1H),7.30(d,J=7.6Hz,1H),7.20-7.25(m,3H),7.17(d,J=7.6Hz,1H),3.76(t,J=7.2Hz,2H),2.69(t,J=7.6Hz,2H),2.50(s,3H),2.01-2.08(m,2H)。

step 2: deprotection of 2- (3- (3- ((4- (methylthio) phenyl) ethynyl) phenyl) propyl) isoindoline-1, 3-dione, and purification and conversion to the free base as described for example 61 gave example 94 as a light yellow solid. Yield (0.050g, 45%):¹HNMR(400MHz,CDCl₃)7.44(d,J=8.4Hz,2H),7.34-7.36(m,2H),7.15-7.28(m,4H),2.74(t,J=7.2Hz,2H),2.66(t,J=7.6Hz,2H),2.50(s,3H),1.75-1.83(m,2H)。

example 95

Preparation of 2- (3- (thien-2-ylethynyl) phenoxy) ethylamine

2- (3- (thien-2-ylethynyl) phenoxy) ethylamine was prepared according to the method described in example 62.

Step 1: alkyne 56 is coupled with 2-bromothiophene and purified by flash chromatography (5-20% ethyl acetate-hexanes) to give tert-butyl 2- (3- (thiophen-2-ylethynyl) phenoxy) ethylcarbamate as a yellow oil. Yield (0.605g, 57%):¹HNMR(400MHz,CDCl₃)7.30(dd,J=5.2,1.2Hz,1H),7.28-7.29(m,1H),7.25(d,J=8.4Hz,1H),7.12(dt,J=7.6,1.2Hz,1H),7.03-7.04(m,1H),7.02(dd,J=5.2,3.6Hz,1H),6.89(ddd,J=8.0,2.4,0.8Hz,1H),4.99(brs,1H),4.04(t,J=4.8Hz,2H),3.55(dt,J=5.2,4.8Hz,2H),1.46(s,9H)。

step 2: tert-butyl 2- (3- (thien-2-ylethynyl) phenoxy) ethylcarbamate was added according to the method described in example 62Deprotection is carried out. After completion of the reaction, diethyl ether was added and the solid was collected by filtration and dried under vacuum to give example 95 hydrochloride salt as a white solid. Yield (0.436g, 88%): ¹HNMR(400MHz,DMSO-d₆)8.14(brs,3H),7.66(dd,J=5.2,1.2Hz,1H),7.40(dd,J=3.6,1.2Hz,1H),7.14(dd,J=7.6,1.2Hz,1H),7.10-7.12(m,3H),7.03(ddd,J=8.4,2.4,1.2Hz,1H),4.19(t,J=5.2Hz,2H),3.17(t,J=5.2,2H)。

Example 96

Preparation of 2- (3- (thien-3-ylethynyl) phenoxy) ethylamine

2- (3- (thien-3-ylethynyl) phenoxy) ethylamine was prepared according to the method described in example 62.

Step 1: alkyne 56 was coupled with 3-bromothiophene and purified by flash chromatography (13% ethyl acetate-hexanes) to give tert-butyl 2- (3- (thiophen-3-ylethynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.630g, 60%).

Step 2: tert-butyl 2- (3- (thien-3-ylethynyl) phenoxy) ethylcarbamate was deprotected as described in example 62, except that trituration was performed with diethyl ether instead of hexane. Example 96 hydrochloride salt was isolated as an off-white solid. Yield (0.430g, 83%):¹HNMR(400MHz,CDCl₃)8.18(brs,3H),7.87(dd,J=2.8,1.2Hz,1H),7.63(dd,J=4.8,2.8Hz,1H),7.33(t,J=8.0Hz,1H),7.23(dd,J=5.2,1.2Hz,1H),7.08-7.13(m,2H),6.99-7.02(m,1H),4.19(t,J=4.8Hz,2H),3.17(t,J=4.8Hz,2H)。

example 97

Preparation of 2- (3- (pyridin-4-ylethynyl) phenoxy) ethylamine

2- (3- (pyridin-4-ylethynyl) phenoxy) ethylamine was prepared according to the method described in example 96.

Step 1: alkyne 56 was coupled with 4-bromopyridine and purified by flash chromatography (12% ethyl acetate-hexanes) to give tert-butyl 2- (3- (pyridin-4-ylethynyl) phenoxy) ethylcarbamate as a brown oil. Yield (0.298g, 38%):¹HNMR(400MHz,CDCl₃)8.61(d,J=6.0Hz,2H),7.38(d,J=6.0Hz,2H),7.29(t,J=8.0Hz,1H),7.17(d,J=8.0Hz,1H),7.07(brs,1H),6.94(ddd,J=8.4,2.4,0.8Hz,1H),4.99(brs,1H),4.05(t,J=5.2,2H),3.55-3.57(m,2H),1.46(s,9H)。

step 2: deprotection of tert-butyl 2- (3- (thien-3-ylethynyl) phenoxy) ethylcarbamate afforded example 97 hydrochloride as a white solid. Yield (0.298g, 38%): ¹HNMR(400MHz,DMSO-d₆)8.78(d,J=1.0Hz,2H),8.23(brs,3H),7.85(s,2H),7.41(t,J=8.0Hz,1H),7.25-7.27(m,2H),7.13(dd,J=8.0,2.8Hz,1H),4.22(t,J=5.2Hz,2H),3.18(q,J=5.2Hz,2H)。

Example 98

Preparation of 2- (3- (pyridin-2-ylethynyl) phenoxy) ethylamine

2- (3- (pyridin-2-ylethynyl) phenoxy) ethylamine was prepared according to the method described in example 96.

Step 1: alkyne 56 was coupled with 4-bromopyridine and purified by flash chromatography (17% ethyl acetate-hexanes) to give tert-butyl 2- (3- (pyridin-2-ylethynyl) phenoxy) ethylcarbamate as a yellow oil. Yield (0.50g, 64%):¹HNMR(400MHz,CDCl₃):8.63(d,J=4.0Hz,1H),7.69(dt,J=7.6,1.6Hz,1H),7.53(d,J=7.6Hz,1H),7.24-7.26(m,2H),7.21(dt,J=8.0,1.2Hz,1H),7.12-7.13(m,1H),6.93(ddd,J=8.0,2.4,1.2Hz,1H),4.98(brs,1H),4.03(t,J=5.2,2H),3.54-3.56(m,2H),1.46(s,9H)。

step 2: deprotection of tert-butyl 2- (3- (pyridin-2-ylethynyl) phenoxy) ethylcarbamate afforded example 98 hydrochloride as a white solid. Yield (0.300g, 85%):¹HNMR(400MHz,DMSO-d₆)8.61(dt,J=5.2,0.8Hz,1H),8.20(brs,3H),7.92(dt,J=8.0,2.0Hz,1H),7.69(d,J=8.0Hz,1H),7.47(ddd,J=7.6,5.2,1.2Hz,1H),7.39(t,J=8.0Hz,1H),7.22(d,J=7.6Hz,1H),7.19-7.20(m,1H),7.08(ddd,J=8.0,2.4,0.8Hz,1H),4.21(t,J=5.2Hz,2H),3.18(dt,J=5.6,5.2Hz,2H)。

example 99

Preparation of 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclohexanol

1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclohexanol was prepared according to the procedure used in modified example 19.

Step 1: aryl bromide 25 was coupled with 1-ethynylcyclohexanol according to the coupling conditions described in example 17 and purified by flash chromatography (40% ethyl acetate-hexanes) to give 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide as a pale yellow oil. Yield (0.621g, 55%):¹HNMR(400MHz,DMSO-d₆)9.32(t,J=4.8Hz,1H),7.34(s,1H),7.27-7.30(m,2H),7.22-7.25(m,1H),5.36-5.37(m,2H),4.54-4.58(m,1H),3.19-3.26(m,2H),1.70-1.83(m,4H),1.60-1.63(m,2H),1.45-1.54(m,5H),1.20-1.22(m,1H)。

step 2: except that the solvent was 90% methanol-water and the reaction was stirred overnight to In addition, 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide was deprotected as described in example 18. The reaction mixture was concentrated under reduced pressure and separated in ethyl acetate and water. Na for mixed organic matter₂SO₄Dried and concentrated under reduced pressure. The crude product was crystallized from hot ethyl acetate. After cooling, the product was collected by filtration, washed with ethyl acetate and hexane, and dried. Example 99 was isolated as a white solid. Yield (0.210g, 47%):¹HNMR(400MHz,DMSO-d₆)7.23-7.26(m,4H),5.45(brs,1H),4.68(t,J=8.4Hz,1H),2.62(brs,2H),1.83-1.88(m,2H),1.48-1.67(m,9H),1.15-1.34(m,1H)。

example 100

Preparation of (R) -4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) hept-4-ol

(R) -4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) hept-4-ol is prepared according to the method described in reaction formula 15.

Reaction formula 15

Step 1: to 4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) hept-4-ol (example 19) (1.39g, 4.80mmol) in CH₂Cl₂To a solution (25mL) was added diisopropylethylamine (1.2mL), and 9-fluorenylmethyloxycarbonyl chloride (Fmoc-Cl, 1.35g, 5.2mmol) in CH₂Cl₂(10mL) of the solution. The reaction was stirred at room temperature for 20 minutes, then concentrated under reduced pressure and purified by flash chromatography (30-70% etoac-hexanes gradient) to give alcohol 59. Yield (1.81g, 74%): ¹HNMR(400MHz,DMSO-d₆)7.91(d,J=7.0Hz,2H),7.67(d,J=7.5Hz,2H),7.28-7.43(m,8H),5.19(s,1H),4.80(dd,J=9.1,5.0Hz,1H),4.54(d,J=4.6Hz,2H),4.31(t,J=5.1Hz,1H),3.42-3.53(m,2H),1.99-2.06(m,1H),1.45-1.63(m,4H),1.30-1.32(m,10H),0.92(t,J=7.0Hz,6H)。

Step 2: to alcohol 59(1.81g,3.54mmol) in CH₂Cl₂(25mL) solution NaHCO was added₃(0.890g,10.6mmol) and Dess-Martin oxidant (Dess-Martinperiodinane) (60, 1.65g, 3.9 mmol). The reaction mixture was stirred at room temperature for 10 minutes, then filtered through celite to remove solids. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (20-70% etoac-hexanes gradient) to afford ketone 61. Yield (1.36g, 75%).

And step 3: preparation of (-) -B-chlorodiisopinocampheylborane solution ((-) -DIP-Cl): to an ice-cold solution of (-) - α -pinene (7.42g, 54.56mmol) in hexane (5mL) under argon was added chloroborane-dimethylsulfide complex (2.55mL,24.46mmol) over a period of 1.5 minutes. The mixture was stirred for 2.5 minutes and then warmed to room temperature over a period of 3 minutes. The reaction mixture was heated to 30 ℃ for 2.5 hours. The resulting solution was approximately 1.5M.

To a solution of ketone 61(0.6472g,1.27mmol) in THF (5mL) at-25 deg.C was added diisopropylethylamine (0.0408g,0.32mmol), and (-) -DIP-Cl solution (1.5mL of 1.5M in hexane, 2.25 mmol). The reaction mixture was stirred at-25 ℃ for 12 minutes, then heated to 0 ℃ and stirred for 1 hour 15 minutes. The reaction mixture was warmed to room temperature over 20 minutes, then additional (-) -DIP-Cl solution (1.5mL of 1.5M hexane solution, 2.25mmol) was added. Stirring was continued at room temperature for 35 min, then diisopropylethylamine (1mL, 5.74mmol) and saturated NaHCO were added ₃An aqueous solution. The layers were separated and the aqueous layer was extracted with EtOAc. The combined organics were washed with brine, over MgSO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (10-70% etoac-hexanes gradient) afforded alcohol 62. (yield 0.3295g, 51%).

And 4, step 4: to a solution of alcohol 62(0.3295g,0.644mmol) in THF (6mL) was added1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (DBU, 0.11mL, 0.74 mmol). The mixture was stirred at room temperature for 20 minutes and then concentrated under reduced pressure. Purification by flash chromatography (7M ammonia in methanol/ethyl acetate/hexane 10:40: 50-20: 80: 0) afforded example 100 as an oil. Yield (0.1383g, 74%):¹the HNMR data are consistent with the data reported above for example 19. Chiral HPLC (25 ℃ C.; eluent 90% heptane-ethanol with 0.1% ethanesulfonic acid): 95.5% major enantiomer (AUC), t_R=17.622min (minor enantiomer: 4.4%, t)_R=21.756min)。[α]D = +20.09(26.6 ℃, c =0.980g/100mL in ethanol).

Absolute stereochemistry determination of example 100

The absolute stereochemistry of example 100 was determined by the method described in equation 16, where example 100 and (R) -3-amino-1-phenylpropan-1-ol were synthesized from a common intermediate (bromide 64). The optical rotation of (R) -3-amino-1-phenylpropan-1-ol is consistent with the values reported in the literature (Kamal, Ahmed et al, Tetrahedron: Asymmetry2002,13(18), 2039-52); [ alpha ] to ]_D= 41.8(30 ℃, c =1.0g/100mL in methanol).

Reaction formula 16

Step 1: to aryl bromide 25(1.0552g,3.23mmol) in CH₂Cl₂To the solution (25mL) was added pyridinium chlorochromate (0.9152g,4.2mmol) and celite (1.96 g). The reaction mixture was stirred at room temperature for 1 hour 50 minutes, then a second portion of pyridinium chlorochromate (0.4936g,2.3mmol) was added. Stirring was continued for 1 hour, then the solid was removed by filtration through celite. The filtrate was concentrated under reduced pressure and purified by flash chromatography (10-50% etoac-hexanes gradient) to afford ketone 63 as a white solid. Yield (0.6465g, 62%):¹HNMR(400MHz,DMSO-d₆)9.40(brs,1H),8.06(t,J=2.0Hz,1H),7.93(d,J=7.6Hz,1H),7.83(ddd,J=7.6,2.0,0.8Hz,1H),7.48(t,J=8.0Hz,1H),3.50(t,J=6.8Hz,2H),3.30(t,J=6.8Hz,2H)。

step 2: to an ice-cold solution of ketone 63(0.6465g,1.99mmol) in THF (10mL) was added diisopropylethylamine (0.1mL,0.57mmol), along with freshly prepared (-) -DIP-Cl (2.5mL of 1.67M in hexane, 4.2 mmol). The reaction was warmed to room temperature and stirred for 2.5 hours. Additional (-) -DIP-Cl solution (1mL,1.67mmol) was added and the mixture was stirred for 2.5 hours. The reaction was washed with saturated NaHCO₃Aqueous solution and ethyl acetate. The combined organics were washed with brine and Na₂SO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (10-100% etoac-hexanes gradient) gave aryl bromide 64. Yield (0.62g, 95%): ¹HNMR(400MHz,CDCl₃)7.50(t,J=1.6Hz,1H),7.43(dt,J=7.2,2.0Hz,1H),7.21-7.27(m,2H),4.84(dt,J=8.8,3.2Hz,1H),3.65-3.73(m,1H),3.36-3.43(m,1H),2.47(dd,J=2.9,1.0Hz,1H),1.80-2.00(m,2H)。

And step 3: to a solution of aryl bromide 64(0.2732g, 0.84mmol) in triethylamine (1mL) and DMF (5mL) were added 4-ethynylhept-4-ol (0.35g,2.5mmol), tri (o-tolyl) phosphine (0.0164g,0.05mmol), CuI (0.0110g,0.058mmol) and PdCl₂(Ph₃P)₂(0.0208g,0.03mmol) and the mixture was degassed (vacuum/argon purge three times). The mixture was heated at 60 ℃ for 15 hours and then cooled to room temperature. The mixture was concentrated under reduced pressure and triturated with ethyl acetate. The solid was removed by filtration, and the filtrate was concentrated under reduced pressure. Purification by flash chromatography (10-70% etoac-hexanes gradient) afforded (R) -2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide. Yield (0.2551g, 79%):¹HNMR data are consistent with the above reported (R/S) -2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (intermediate in the synthesis of example 19).

And 4, step 4: to (R) -2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetylTo a solution of amine (0.2551g, 0.66mmol) in methanol-water (2:1, 12mL) was added K₂CO₃(0.4967g,3.6mmol) and the mixture was heated at 60 ℃ for 1 hour. After cooling to room temperature, the mixture was separated in ethyl acetate and brine. Na for mixed organic matter ₂SO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (50:40:10 to 0:80:20 gradient hexanes: ethyl acetate: 7M ammonia in methanol) afforded alkyne 65 as an oil. Yield (0.1421g, 74%):¹the HNMR data are consistent with the data reported above for example 100. Chiral HPLC (25 ℃ C.; eluent 90% heptane-ethanol with 0.1% ethanesulfonic acid): 93.6% Main enantiomer (AUC), t_R=15.880min (minor enantiomer: 6.4%, t)_R=20.068min)。[α]_D= 20.77(24.1 ℃, c =1.920g/100mL in ethanol).

Preparation of (R) -3-amino-1-phenylpropan-1-ol (67) from aryl bromide 64:

step 1: to a solution of aryl bromide 64(1.3155g,4.03mmol) in THF (13mL) at-78 deg.C was added a solution of n-butyllithium (13mL of a 1.6M solution in hexane, 20.8 mmol). The mixture was stirred at-78 ℃ for 10 minutes and then with 30% NH₄The reaction was quenched with aqueous Cl. After warming to room temperature, the mixture was extracted with ethyl acetate. The combined organics were washed with brine, over MgSO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (10-70% ethyl acetate-hexanes) yielded alcohol 66 contaminated with-3% bromide starting material. Yield (0.3730g, 37%):¹HNMR(400MHz,CDCl₃)9.33(brs,1H),7.51(t,J=7.6Hz,1H),7.44(d,J=8.0Hz,1H),7.39(s,1H),7.33(ddd,J=8.4,2.8,0.8Hz,1H),5.59(d,J=4.8Hz,1H),4.66(dt,J=8.0,4.4Hz,1H),3.19-3.28(m,2H),1.71-1.86(m,2H)。

step 2: deprotection of alcohol 66 and purification as described for alkyne 65 gave (R) -3-amino-1-phenylpropan-1-ol (67) as an oil. Yield (0.1749g, 77%): ¹HNMR(400MHz,CDCl₃)7.31-7.38(m,4H),7.21-7.25(m,1H),4.94(dd,J=8.8,3.2Hz,1H),3.05-3.10(m,1H),2.91-2.97(m,1H),2.62(brs,3H),1.82-1.89(m,1H),1.70-1.79(m,1H)。[α]_D=+34.84(25.7℃，c=2.05g/100mL，In methanol).

Example 101

Preparation of (S) -4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) hept-4-ol

(S) -4- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure described for example 100.

Step 1: reduction of ketone 61 with (+) -DIP-Cl and purification by flash chromatography (10-70% etoac-hexanes gradient) afforded (S) - (9H-fluoren-9-yl) methyl 3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propylcarbamate. Yield (0.50g, 70%):¹HNMR data are consistent with those reported above.

Step 2: 3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propylcarbamic acid (S) - (9H-fluoren-9-yl) methyl ester was deprotected and purified according to the method described for example 100 to give example 101 as an oil. Yield (0.1845, 65%):¹the HNMR data are consistent with the reported data for example 19. Chiral HPLC (25 ℃ C.; eluent 90% heptane-ethanol with 0.1% ethanesulfonic acid): 95.7% major enantiomer (AUC), t_R=21.562min (minor enantiomer: 4.2%, t)_R=17.572min)。[α]_D= 24.93(26.6 ℃, c =0.955g/100mL in ethanol).

Example 102

Preparation of 3- (3- (hept-1-ynyl) phenyl) propan-1-amine

3- (3- (hept-1-ynyl) phenyl) propan-1-amine was prepared according to the procedure used in example 36.

Step 1: aryl bromide 34 was coupled with hept-1-yne and purified by flash chromatography (30% ethyl acetate-hexanes) to give tert-butyl 3- (3- (hept-1-ynyl) phenyl) propylcarbamate as a yellow oil. Yield (0.300g, 57%).

Step 2: t-butyl 3- (3- (hept-1-ynyl) phenyl) propylcarbamate was deprotected as described in example 36. Purification by preparative HPLC (method 004P) gave example 102 hydrochloride as a white solid. Yield (0.200g, 15%):¹HNMR(400MHz,DMSO-d₆)7.98(brs,2H),7.28(t,J=7.6Hz,1H),7.24-7.18(m,3H),2.74(t,J=7.2Hz,2H),2.63(t,J=7.6Hz,2H),2.41(t,J=6.8Hz,2H),1.84(quint,J=7.6Hz,2H),1.54(quint,J=7.6Hz,2H),1.43-1.28(m,4H),0.89(t,J=7.2Hz,3H)。

example 103

Preparation of N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide

N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide was prepared according to the procedure described for example 60.

Acylation of 4- ((3- (3-aminopropyl) phenyl) ethynyl) heptan-4-ol (example 2) gave example 103 as a clear oil. Yield (0.087g, 80%):¹HNMR(400MHz,DMSO-d₆)7.81(bs,1H),7.24(t,J=7.6Hz,1H),7.12-7.15(m,3H),5.11(s,1H),2.99(q,J=12.8,7.8Hz,2H),2.54(t,J=7.2,2H),1.77(s,3H),1.68-1.42(m,10H),0.89(t,J=6.8Hz,6H)。

example 104

Preparation of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide

2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide (compound 26) was prepared according to the procedure described for example 19, scheme 5.

Example 105

Preparation of 3- (3- (cycloheptylethynyl) phenyl) propan-1-amine

3- (3- (cycloheptylethynyl) phenyl) propan-1-amine is prepared according to the procedure described in scheme 17.

Reaction formula 17

Step 1: LAH (180mL of a 1M solution, 176mmol) was slowly added to a solution of acid 68(25g,176mmol) in dry THF (500mL) at 5 ℃ under argon. The reaction was warmed to room temperature, stirred for 1 hour, cooled again to 5 ℃ and then quenched by the slow addition of saturated Na₂SO₄The reaction was quenched with aqueous solution. The resulting precipitate was removed by filtration, and the filtrate was extracted with ethyl acetate, washed with water and brine, and washed with Na₂SO₄Dry, filter and concentrate to give alcohol 69 as a colorless oil. Yield (21g, 98%).¹HNMR(400MHz,CDCl₃)3.38(d,J=6.4Hz,2H),1.38–1.80(m,12H),1.10–1.20(m,2H)。

Step 2: a solution of alcohol 69(4.5g,35mmol) in dichloromethane (10mL) was added to a stirred mixture of pyridinium chlorochromate (9.4g,43.8mmol) and diatomaceous earth (10g) in dichloromethane (100mL) and the reaction stirred for 16 h. The mixture was filtered through a pad of silica gel and the pad was rinsed with diethyl ether. The combined filtrates were concentrated to give impure aldehyde 70 as a green oil which was taken to the next step without purification. Yield (4.1g, 93%).¹HNMR(400MHz,CDCl₃)9.58(d,J=0.8Hz,1H),2.28–2.36(m,1H),1.88–1.95(m,2H),1.40–1.70(m,10H)。

And step 3: sodium trichloroacetate was added in 3 portions over 10 minutes to a stirred solution of aldehyde 70(14.9g,118mmol) and trichloroacetic acid (19.3g,177mmol) in DMF (150 mL). The reaction was stirred at room temperature for 2 hours, cooled with an ice bath, then quenched with water and diluted. The solution was extracted with hexane and saturated NH ₄Aqueous Cl, water and brine. Mixing the organic substances with Na₂SO₄Drying, filtration and concentration under reduced pressure gave the impure alcohol 71 as a yellow oil which was carried on to the next step without purification. Yield (23.4g, 80%).¹HNMR(400MHz,CDCl₃)3.95(d,J=2.0Hz,1H),2.85(brs,1H),2.20–2.30(m,1H),1.88–2.08(m,1H),1.36–1.84(m,11H)。

And 4, step 4: p-toluenesulfonylchloride (3.34g,17.5mmol) was added to a solution of alcohol 71(4.3g,17.5mmol), triethylamine (3.6mL,26.3mmol) and diazabicyclooctane (0.586g,5.2mmol) in 40mL of dichloromethane and stirred at room temperature for 90 min. The reaction was quenched by washing with water (40mL) followed by 5N HCl. The combined aqueous solutions were extracted with dichloromethane (40mL) and the combined organics were further washed with 2N HCl, water, and brine, then Na₂SO₄Dried, filtered, and concentrated under reduced pressure. Purification by flash chromatography (0-10% etoac/hexanes gradient) afforded sulfonate 72 as a pale yellow crystal. Yield (2.65g, 38%):¹HNMR(400MHz,CDCl₃)7.81(d,J=8.8Hz,2H),7.32(d,J=8Hz,2H),2.43(s,3H),2.24–2.32(m,1H),1.98–2.06(s,1H),1.78–1.88(m,1H),1.60–1.73(m,3H),1.28–1.60(m,8H)。

and 5: methyllithium (7.0mL of a 1.6M solution in diethyl ether, 11.25mmol) was added dropwise to a stirred solution of the sulfonate 72(1g, 2.5mmol) in anhydrous THF (15mL) at 5 deg.C under argon. The reaction was warmed to room temperature, stirred for 16 hours, and then quenched by the slow addition of saturated NH₄The reaction was quenched with aqueous Cl. The mixture was extracted with hexane and the combined organics were washed with brine, Na ₂SO₄Drying, filtration, and concentration under reduced pressure gave alkyne 73 as a yellow oil. Yield (0.270g, 88%):¹HNMR(400MHz,CDCl₃)2.51(s,1H),1.99(d,J=2.8Hz,1H),1.73–1.83(m,2H),1.55–1.70(m,4H),1.35–1.55(m,6H)。

step 6: sonogashira coupling was performed as described in example 1. Purification by flash chromatography (0-25% etoac-hexanes gradient) afforded alkyne 74 as an amber oil. Yield (0.556g, 59%):¹HNMR(400MHz,CDCl₃)7.16–7.26(m,3H),7.03–7.08(m,1H),6.28(brs,1H),3.36(dt,J=6.8,6.8Hz,2H),2.75–2.83(m,1H),2.63(t,J=7.2Hz,2H),1.85–1.95(m,4H),1.69–1.80(m,4H),1.48–1.65(m,6H)。

and 7: the alkyne 74 is deprotected as described in example 1. Purification by flash chromatography (5% (7N ammonia/methanol)/dichloromethane) gave example 106 as a colorless oil. Yield (0.226g, 56%):¹HNMR(400MHz,CDCl₃)7.13–7.25(m,3H),7.03–7.08(m,1H),2.74–2.82(m,1H),2.69(t,J=7.2Hz,2H),2.59(t,J=7.2Hz,2H),1.84-1.94(m,2H),1.68–1.80(m,6H),1.46–1.64(m,6H),1.30(brs,2H)。

example 106

Preparation of 2- (3- (cycloheptylethynyl) phenoxy) ethylamine

2- (3- (cycloheptylethynyl) phenoxy) ethylamine was prepared as described in example 18.

Step 1: the alkyne 73 is Sonogashira coupled with aryl bromide 19 and then purified by flash chromatography (0-25% etoac-hexanes gradient) to give N- (2- (3- (cycloheptylethynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide as an amber oil. Yield (0.154g, 27%):¹HNMR(400MHz,CDCl₃)7.12(t,J=8Hz,1H),6.98–6.98(m,1H),6.83–6.86(m,1H),6.65–6.80(m,2H),4.01(t,J=4.8Hz,2H),3.66–3.73(m,2H),2.68–2.77(m,1H),1.79–1.88(m,2H),1.63–1.73(m,4H),1.40–1.57(m,6H)。

step 2: deprotection of N- (2- (3- (cycloheptylethynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide followed by flash chromatography (5% (7N ammonia/methanol)/dichloromethane) gave example 106 as a light brown oil. Yield (0.064g, 53%): ¹HNMR(400MHz,CDCl₃)7.16(t,J=8Hz,1H),6.95–6.99(m,1H),6.90–6.94(m,1H),6.78–6.83(m,1H),3.95(t,J=5.2Hz,2H),3.05(t,J=4.8Hz,2H),2.74–2.82(m,1H),1.84–1.94(m,2H),1.68–1.80(m,4H),1.44–1.64(m,8H)。

Example 107

Preparation of 3-amino-1- (3- (3-methoxyprop-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (3-methoxyprop-1-ynyl) phenyl) propan-1-ol is prepared according to the procedure described in equation 18.

Reaction formula 18

Step 1: to THF (200mL) cooled to-50 deg.C) And CH₃CN (7.8mL,148mmol) was added portionwise to the stirred mixture KOtBu (18.16g,162mmol) and the temperature was maintained at-50 ℃. After stirring for 30 minutes, 3-bromobenzaldehyde (25g,135mmol) was added while maintaining the same internal temperature. After stirring for 30 minutes, the reaction mixture was heated to 0 ℃ and stirred for an additional 3 hours. It was cooled again to-10 ℃ and the reaction quenched with excess water. Extraction with ethyl acetate gave a crude product which was purified by flash chromatography (0-20% etoac-hexanes gradient) to give 23 as a pale yellow oil. Yield (21g, 69%):¹HNMR(400MHz,DMSO-d₆)7.61(s,1H),7.49(bd,J=7.8Hz,1H),7.47(bd,J=7.6Hz,1H),7.34(t,J=7.6Hz,1H),6.09(d,J=4.4Hz,1H),4.92(m,1H),2.81-2.95(m,2H)。

step 2: to a solution of nitrile 23(22.4g,99mmol) in dry THF (200mL) under nitrogen was added BH via an additional funnel₃.SMe₂(28.4mL,297mmol) and the addition time was 1 hour. The mixture was then refluxed for 14 hours. After cooling to 0 ℃, the reaction was quenched by the slow addition of excess borane to methanol. It was concentrated to dryness under reduced pressure. This step was repeated six times. The crude product was then dissolved in 6n hcl and extracted with DCM. The aqueous layer was basified with concentrated aqueous ammonia to pH 10 and extracted with DCM. Mixing the organic substances with Na ₂SO₄And (5) drying. The solution was filtered and concentrated under reduced pressure to give 24 as a clear oil. Yield (15.94g, 70%):¹HNMR(400MHz,DMSO-d₆)7.51(s,1H),7.42(bd,J=7.6Hz,1H),7.22-7.34(m,2H),4.69(m,1H),2.67-2.74(m,2H),1.65-1.77(m,2H)。

and step 3: to a solution of 24(15.9g,69mmol) in anhydrous THF (200mL) was added ethyl trifluoroacetate (10mL,83 mmol). The reaction mixture was stirred at room temperature for 3 hours, during which time the conversion process was complete. The reaction mixture was then concentrated to dryness under reduced pressure. The product is sufficiently pure to be used in the next conversion step. Yield (19g, crude):¹HNMR(400MHz,DMSO-d₆)9.35(bs,1H),7.53(s,1H),7.44(bd,J=7.6Hz,1H),7.22-7.34(m,2H),5.48(d,J=4.8Hz,1H),4.57-45.61(m,1H),3.18-3.30(m,2H),1.73-1.87(m,2H)。

and 4, step 4: 25, methyl propargyl ether (0.2mL,2.25mmol), PdCl₂(PPh₃)₂A mixture of (108mg,0.075mmol), tri-o-tolylphosphine (47mg,0.075mmol), cuprous iodide (29mg,0.075mmol) in diisopropylamine (10mL) was heated at reflux overnight. The mixture was cooled to room temperature and then concentrated under reduced pressure. Purification by flash chromatography (0-30% etoac-hexanes gradient) afforded ether 76 as a yellow oil. Yield (0.4g, 82%):¹HNMR(400MHz,CDCl₃)7.44(s,1H),7.38-7.40(m,1H),7.31-7.34(m,2H),4.88(m,1H),4.12(s,2H),3.66-3.70(m,1H),3.43(s,3H),3.38-3.41(m,1H),2.37(d,J=2Hz,1H)1.93-1.97(m,2H)。

and 5: a mixture of 76, potassium carbonate (438mg,3.26mmol) and water (2mL) in 2-PrOH (10mL) was heated at reflux overnight. The reaction mass was concentrated to dryness under reduced pressure and diluted with water (10 mL). This material was acidified to pH 2 and extracted with DCM. The aqueous layer was washed with saturated NaHCO₃The solution was basified to pH 10 and extracted with DCM. Mixing the organic substances with Na ₂SO₄And (5) drying. The solution was concentrated under reduced pressure to give example 107 as a brown oil. Yield (0.138g, 77%):¹HNMR(400MHz,DMSO-d₆)7.40(s,1H),7.25-7.35(m,3H),4.67(t,J=6.4Hz,1H),4.34(s,2H),2.59-2.64(m,2H),2.50(s,3H),1.74-1.80(m,2H)。¹³CNMR(100MHz,DMSO-d₆)147.1,129.5,128.7,128.4,126.2,121.5,86.1,85.5,70.8,59.5,57.0,42.2。ESIMSm/z220[M+1]⁺。

example 108

Preparation of 3-amino-1- (3- (hex-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (hex-1-ynyl) phenyl) propan-1-ol was prepared as described for example 107.

Step 1: sonogashira reaction of 25 with 1-hexyne gave 2,2, 2-trifluoro-N- (3- (3- (hex-1-ynyl) phenyl) -3-hydroxypropyl) acetamide as a yellow oil. Yield (1.53g, 76%):¹HNMR(400MHz,CDCl₃)7.38(s,1H),7.25-7.34(m,3H),4.88(m,1H),3.65-3.73(m,1H),3.38-3.42(m,1H),2.40(t,J=7.2Hz,2H),2.25(d,J=2.0,1H)1.93-1.99(m,2H),1.45-1.61(m,4H),0.94(t,J=7.2Hz,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- (hex-1-ynyl) phenyl) -3-hydroxypropyl) acetamide gave a yellow oil. The crude product was dissolved in methanol (5mL) and stirred with HCl in dioxane (1mL,4M) for 30 minutes. The mixture was concentrated to dryness under reduced pressure. Purification by flash chromatography afforded example 108 as the hydrochloride salt as a pale yellow semi-solid. Yield (0.17g, 41%):¹HNMR(400MHz,DMSO-d₆)7.23-7.32(m,4H),4.62-4.65(m,1H),2.78-2.86(m,2H),2.38(t,J=6.8Hz,2H),1.74-1.85(m,2H),1.35-1.52(m,4H),0.87(t,J=7.2Hz,3H)。¹³CNMR(100MHz,DMSO-d₆)145.7,129.7,128.5,128.4,125.2,123.1,90.5,80.7,69.2,36.5,36.3,30.3,21.5,18.3,13.5。ESIMSm/z232[M+1]⁺。

example 109

Preparation of 4- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) hept-4-ol

4- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure described in equation 19.

Reaction formula 19

Step 1: under nitrogenTo a solution of 1- (3-bromophenyl) -3-chloroprop-2-ol (8.49g,34.0mmol) in anhydrous DMF (100mL) was added NaN ₃(11.05g,170.0mmol) and NaI (cat.,0.75g,5.0 mmol). The mixture was heated to 75 ℃ overnight. After cooling to room temperature, the mixture was diluted with ether and washed with water and brine. Adding Na to the solution₂SO₄Dried and concentrated under reduced pressure. The product was dried in a vacuum oven at 40 ℃ for 2 hours to give 1-azido-3- (3-bromophenyl) propan-2-ol as a yellow oil, which was used without purification. Yield (8.6g, 98% crude).

Step 2: to a solution of 1-azido-3- (3-bromophenyl) propan-2-ol (8.59g,33.28mmol) in THF (60mL) under nitrogen was added PPh₃(8.73g,33.28mmol) and water (20 mL). The reaction mixture was heated to 50 ℃ for 24 hours. After cooling to room temperature, the mixture was diluted with brine and extracted with ethyl acetate. Na for organic layer₂SO₄Dried and concentrated under reduced pressure. The crude amine was dissolved in THF (20ml) and ethyl trifluoroacetate (20ml) and stirred at room temperature overnight. Evaporation under reduced pressure followed by purification by flash chromatography (5% ethyl acetate in dichloromethane) gave bromide 77 as a white solid. Yield (3.72g, 35%):¹HNMR(400MHz,DMSO-d₆)9.33(t,J=5.2Hz,1H),7.42(d,J=1.2Hz,1H),7.38-7.35(m,1H),7.24-7.20(m,2H),5.00(d,J=6.0Hz,1H),3.83-3.75(m,1H),3.21-3.07(m,2H),2.70(dd,J=13.6Hz,4.8,1H),2.55(dd,J=14.0,6.0Hz,1H)。

and step 3: bromide 77 was coupled with 4-ethynylhept-4-ol as described in example 17 to give 2,2, 2-trifluoro-N- (2-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide as a clear oil. Yield (0.455g, 59%): ¹HNMR(400MHz,DMSO-d₆)9.32(t,J=5.4Hz,1H),7.25-7.22(m,2H),7.18-7.16(m,2H),5.11(s,1H),4.96(d,J=5.6Hz,1H),3.81-3.74(m,1H),3.21-3.07(m,2H),2.68(dd,J=14.0,4.6Hz,1H),2.54(dd,J=14.0,7.8Hz,1H),1.61-1.40(m,8H),0.89(t,J=7.2Hz,6H)。

And 4, step 4: deprotection of 2,2, 2-trifluoro-N- (2-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propyl) acetamide to remove impurities as described in example 1Example 109 was obtained as a pale yellow oil. Yield (0.273g, 82%):¹HNMR(400MHz,DMSO-d₆)7.24-7.20(m,2H),7.17-7.13(m,2H),5.11(bs,1H),4.55(bs,1H),3.51-3.46(m,1H),2.67(dd,J=13.6,5.2Hz,1H),2.50(dd,J=13.6,7.6Hz,1H),2.45(dd,obs.,1H),2.37(dd,J=12.8,6.8Hz,1H),1.61-1.40(m,8H),0.89(t,J=7.6Hz,6H)。

example 110

Preparation of 1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cyclohexanol

1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cyclohexanol was prepared according to the procedure described for example 109.

Step 1: bromide 77 was coupled with 1-ethynylcyclohexanol to give 2,2, 2-trifluoro-N- (2-hydroxy-3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide as a clear oil. Yield (0.48g, 65%):¹HNMR(400MHz,DMSO-d₆)9.32(t,J=5.6Hz,1H),7.26-7.22(m,2H),7.20-7.16(m,2H),5.37(s,1H),4.97(d,J=5.6Hz,1H),3.82-3.75(m,1H),3.21-3.07(m,2H),2.66(dd,J=14.0,4.8Hz,1H),2.55(dd,J=13.6,7.8Hz,1H),1.83-1.79(m,2H),1.63-1.60(m,2H),1.54-1.44(m,5H),1.23-1.18(m,1H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2-hydroxy-3- (3- ((1-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide afforded example 110 as a light yellow solid. Yield (0.227g, 65%):¹HNMR(400MHz,DMSO-d₆)7.25-7.21(m,2H),7.18-7.15(m,2H),5.37(bs,1H),4.59(bs,1H),3.53-3.47(m,1H),2.67(dd,J=13.6,5.2Hz,1H),2.51(dd,J=13.6,7.6Hz,1H),2.48(obsm,1H),2.38(dd,J=12.8,6.8Hz,1H),1.83-1.77(m,2H),1.63-1.60(m,2H),1.54-1.45(m,5H),1.23-1.18(m,1H)。

example 111

Preparation of 1- (3- (3-amino-2-hydroxypropyl) phenyl) -3-ethylpent-1-yn-3-ol

1- (3- (3-amino-2-hydroxypropyl) phenyl) -3-ethylpent-1-yn-3-ol was prepared according to the method described for example 109.

Step 1: bromide 77 was coupled with 3-ethylpent-1-yn-3-ol to give N- (3- (3- (3-ethyl-3-hydroxypent-1-ynyl) phenyl) -2-hydroxypropyl) -2,2, 2-trifluoroacetamide as a clear oil. Yield (0.472g, 66%): ¹HNMR(400MHz,DMSO-d₆)9.32(t,J=5.6Hz,1H),7.26-7.22(m,2H),7.20-7.16(m,2H),5.11(s,1H),4.96(d,J=5.6Hz,1H),3.80-3.74(m,1H),3.21-3.07(m,2H),2.68(dd,J=14.0,4.8Hz,1H),2.53(dd,J=13.6,7.6Hz,1H),1.66-1.52(m,4H),0.96(t,J=7.2Hz,6H)。

Step 2: deprotection of N- (3- (3- (3-ethyl-3-hydroxypent-1-ynyl) phenyl) -2-hydroxypropyl) -2,2, 2-trifluoroacetamide affords example 111 as a light yellow oil. Yield (0.232g, 69%):¹HNMR(400MHz,DMSO-d₆)7.24-7.20(m,2H),7.17-7.15(m,2H),5.11(bs,1H),4.55(bs,1H),3.51-3.45(m,1H),2.67(dd,J=13.6,5.2Hz,1H),2.51(dd,J=13.6,7.6Hz,1H),2.45(obsdm,J=44Hz,1H),2.37(dd,J=12.8,6.8Hz,1H),1.66-1.52(m,4H),0.96(t,J=7.2Hz,6H)。

example 112

Preparation of 1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cyclopentanol

1- ((3- (3-amino-2-hydroxypropyl) phenyl) ethynyl) cyclopentanol was prepared according to the general method described for example 109.

Step 1: the bromide 77 was coupled with 1-ethynylcyclopentanol to give 2,2, 2-trifluoro-N- (2-hydroxy-3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) propyl) acetamide as a clear oil. Yield (0.441g, 62%):¹HNMR(400MHz,DMSO-d₆)9.32(t,J=5.6Hz,1H),7.26-7.22(m,2H),7.19-7.16(m,2H),5.26(bs,1H),4.97(bs,1H),3.78(bs,1H),3.20-3.06(m,2H),2.67(dd,J=14.0,4.8Hz,1H),2.53(dd,J=13.6,7.6Hz,1H),1.91-1.79(m,4H),1.76-1.60(m,4H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2-hydroxy-3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) propyl) acetamide afforded example 112 as a light yellow solid. Yield (0.217g, 69%):¹HNMR(400MHz,DMSO-d₆)7.24-7.20(m,2H),7.17-7.15(m,2H),5.26(bs,1H),4.55(bs,1H),3.51-3.45(m,1H),2.66(dd,J=13.6,5.2Hz,1H),2.51(dd,J=13.6,8.0Hz,1H),2.45(obsdm,J=44Hz,1H),2.36(dd,J=12.8,6.8Hz,1H),1.91-1.80(m,4H),1.76-1.60(m,4H)。

example 113

Preparation of 2- (3- (cyclopropylethynyl) phenoxy) ethylamine

2- (3- (cyclopropylethynyl) phenoxy) ethylamine was prepared according to the procedure described for example 18.

Step 1: the bromide 19 was coupled with cyclopropylacetylene to give N- (2- (3- (2-cyclopropylethynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide as a clear oil. Yield (2.0g, 71%): the crude product was used directly for further deprotection reactions.

Step 2: deprotection of N- (2- (3- (2-cyclopropylethynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide to give a light yellow oilExample 113 of (a). Yield (0.350g, 52%):¹HNMR(400MHz,DMSO-d₆)7.19-7.24(m,1H),6.87-6.93(m,3H),3.90(t,J=5.6Hz,2H),2.86(t,J=5.6Hz,2H),1.49-1.56(m,1H),0.85-0.90(m,2H),0.72-0.77(m,2H):¹³CNMR(100MHz,DMSO-d₆)158.4,129.6,124.2,123.6,116.7,114.9,93.6,75.5,69.9,40.7,8.3,-0.3.ESIMSm/z202[M+1]⁺。

example 114

Preparation of 5- (3- (2-aminoethoxy) phenyl) pent-4-yn-1-ol

5- (3- (2-Aminoethoxy) phenyl) pent-4-yn-1-ol is prepared as described in example 18.

Step 1: a mixture of bromide (19) (2.5g,8mmol), pentyn-1-ol (1.34g,16mmol) in triethylamine (6mL,60mmol) and DMF (18mL) was purged with nitrogen for 10 min. Then, PdCl was added₂(PPh₃)₂(0.28g,0.4mmol)、P(o-Tol)₃(0.122g,0.4mmol) and CuI (0.076g,0.4mmol), and the flask was again purged with nitrogen and the resulting mixture was heated at 90 ℃ overnight. Then, it was poured into water and extracted with ethyl acetate. The combined organic layers were washed with water and anhydrous Na₂SO₄Dried, filtered and concentrated under reduced pressure. Purification by flash chromatography (0-30% etoac-hexanes gradient) afforded 2,2, 2-trifluoro-N- (2- (3- (5-hydroxypent-1-ynyl) phenoxy) ethyl) acetamide as a yellow oil. Yield (1.61g, 63%):¹HNMR(400MHz,CDCl₃)7.18-7.24(m,1H),7.03(d,J=7.6Hz,1H),6.91(dd,J=0.8,1.2Hz,1H),6.83(dd,J=5.6,2.0Hz,1H),6.76(bs,1H),4.06-4.10(m,2H),3.76-3.84(m,4H),2.53(t,J=7.2Hz,2H),1.82-1.90(m,2H)。

step 2: to a stirred solution of alkyne (1.6g,5mmol) in methanol-water (25mL:5mL) was added K₂CO₃(3.5g,25mmol) and the resulting mixture was stirred overnight. The solvent was removed under reduced pressure. The residue was separated with ethyl acetate and water, and the combined organics were washed with water and Na ₂SO₄And (5) drying. The filtered solution was concentrated under reduced pressure to give example 114 as a yellow oil. Yield (0.360g, 33%):¹HNMR(400MHz,DMSO-d₆)7.21-7.26(m,1H),6.94(s,1H),6.88-6.93(m,2H),4.55(bs,2H),3.90(t,J=6.4Hz,2H),3.50(t,J=6.4Hz,2H),2.84(t,J=5.6Hz,2H),2.44(t,J=7.2Hz,2H),1.63-1.71(m,2H).¹³CNMR(100MHz,DMSO-d₆)159.0,130.1,124.8,124.0,117.1,115.3,90.8,80.8,70.7,59.9,41.3,32.0,15.7.ESIMSm/z220[M+1]⁺。

example 115

Preparation of 5- (3- (3-aminopropyl) phenyl) pent-4-yn-1-ol

5- (3- (3-aminopropyl) phenyl) pent-4-yn-1-ol was prepared according to the method described in example 127.

Step 1: sonogashira coupling of bromide 87(0.5g,1.5mmol) with 4-pentyn-1-ol gave (tert-butyl 3- (5-hydroxypent-1-ynyl) phenylethylcarbamate) (0.35g, 69%):¹HNMR(400MHz,CDCl₃)7.17-7.24(m,3H),7.07-7.11(m,1H),3.83(t,J=6.0Hz,2H),3.13-3.15(m,2H),2.50-2.61(m,4H),2.05(s,1H),1.77-1.89(m,4H),1.44(s,9H)。

step 2: tert-butyl (3- (5-hydroxypent-1-ynyl) phenyl) ethylcarbamate was deprotected with HCl/dioxane in THF to give the hydrochloride salt of example 115. Yield (0.14g, 63%):¹HNMR(400MHz,DMSO-d₆)7.22-7.27(m,1H),7.14-7.21(m,3H),3.48(t,J=6.0Hz,2H),2.75(t,J=7.6Hz,2H),2.57(t,J=7.6Hz,2H),2.40(t,J=6.8Hz,2H),1.74-1.82(m,2H),1.61-1.68(m,2H)。

example 116

Preparation of 2- (3- (hex-1-ynyl) phenoxy) ethylamine

2- (3- (hex-1-ynyl) phenoxy) ethylamine was prepared as described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with 1-hexyne to give 2,2, 2-trifluoro-N- (2- (3- (hex-1-ynyl) phenoxy) ethyl) acetamide as a clear oil. Yield (1.8g, 72%):¹HNMR(400MHz,CDCl₃)7.19-7.23(m,1H),7.05(d,J=7.6Hz,1H),6.84(s,1H),6.80(dd,J=8.0,2.4Hz,1H),4.10(t,J=5.2Hz,2H),3.77-3.80(m,2H),2.40(t,J=7.2Hz,2H),1.53-1.61(m,2H),1.43-1.50(m,2H),0.95(t,J=7.2Hz,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (hex-1-ynyl) phenoxy) ethyl) acetamide gave example 116 as a yellow oil. Yield (0.620g, 90%): ¹HNMR(400MHz,DMSO-d₆)7.20-7.25(m,1H),6.87-6.93(m,3H),3.91(t,J=5.2Hz,2H),2.79-2.87(m,2H),2.38(t,J=6.4Hz,2H),1.42-1.53(m,2H),1.30-1.40(m,2H),0.88(t,J=7.2Hz,3H).¹³CNMR(100MHz,DMSO-d₆)158.9,130.1,124.8,124.1,117.2,115.3,90.9,80.9,70.3,41.1,30.7,21.9,18.7,13.9.ESIMSm/z218[M+1]⁺。

Example 117

Preparation of 2- (3- (3-methoxyprop-1-ynyl) phenoxy) ethylamine

2- (3- (3-Methoxyprop-1-ynyl) phenoxy) ethylamine was prepared according to the method described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with 3-methoxypropyne to give 2,2, 2-trifluoro-N- (2- (3- (3-methoxyprop-1-ynyl) phenoxy) ethyl) acetamide as a clear oil. Yield (0.51g, 21%):¹HNMR(400MHz,CDCl₃)7.22-7.27(m,1H),7.10(d,J=7.6Hz,1H),6.98(s,1H),6.88(dd,J=6.8,1.6Hz,1H),6.71(bs,1H),4.32(s,2H),4.10(t,J=5.2Hz,2H),3.77-3.82(m,2H),3.45(s,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (3-methoxyprop-1-ynyl) phenoxy) ethyl) acetamide gave example 117 as an off-white oil. Yield (0.160g, 47%):¹HNMR(400MHz,DMSO-d₆)7.27-7.32(m,1H),6.96-7.05(m,3H),4.32(s,2H),3.97(t,J=5.6Hz,2H),3.33(s,3H),2.92(t,J=5.6Hz,2H).¹³CNMR(100MHz,DMSO-d₆)158.9,130.4,124.3,123.7,123.4,117.3,116.2,86.2,86.1,69.6,59.9,57.5,40.8.ESIMSm/z206[M+1]⁺。

example 118

Preparation of 3- (3- (3-amino-1-hydroxypropyl) phenyl) prop-2-yn-1-ol

3- (3- (3-amino-1-hydroxypropyl) phenyl) prop-2-yn-1-ol was prepared according to the method described for example 108.

Step 1: sonogashira reaction of 25 with propargyl alcohol to give 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxyprop-1-ynyl) phenyl) propyl) acetamide as a dark yellow oil. Yield (0.37g, 80%):¹HNMR(400MHz,CDCl₃)7.44(s,1H),7.30-7.37(m,3H),4.87(m,1H),4.50(d,J=5.6Hz,2H),3.66-3.69(m,1H),3.40-3.43(m,1H),2.45(bs,1H)1.93-2.04(m,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxyprop-1-ynyl) phenyl) propyl) acetamide (0.48g,1.59mmol) afforded example 118 as the hydrochloride salt as a pale yellow semi-solid. Yield (0.14g, 42%): ¹HNMR(400MHz,DMSO-d₆)8.17(bs,3H),7.29-7.35(m,4H),5.5.59-5.65(bs,1H),5.39(t,J=6.0Hz,1H),4.30(d,J=5.6Hz,2H),2.83(m,2H),1.89(m,2H)。¹³CNMR(100MHz,DMSO-d₆)145.9,130.2,129.8,128.7,126.1,122.5,89.9,84.0,69.5,49.6,36.5,36.4. Mass spectrum: 206[ M +1 ]]⁺。

Example 119

Preparation of 3-amino-1- (3- (4-methylpent-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (4-methylpent-1-ynyl) phenyl) propan-1-ol is prepared as described in example 132.

Step 1: sonogashira reaction of 25 with 4-methylpent-1-yne gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-methylpent-1-ynyl) phenyl) propyl) acetamide as a dark brown oil. Yield (0.94g, 94%):¹HNMR(400MHz,CDCl₃)7.38(s,1H),7.25-7.35(m,3H),4.86(m,1H),3.67-3.72(m,1H),3.38-3.44(m,1H),2.30(d,J=6.4Hz,2H),2.28(bs,1H),1.87-1.99(m,3H),1.05(d,J=6.8Hz,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-methylpent-1-ynyl) phenyl) propyl) acetamide gave example 119 as a yellow oil. Yield (0.508g, 76%):¹HNMR(400MHz,DMSO-d₆)7.33(s,1H),7.23-7.29(m,3H),5.12(bs,2H),4.66(t,J=6.4Hz,1H),2.68(t,J=6.8Hz,2H),2.32(d,J=6.4Hz,2H),1.82-1.86(m,1H),1.67-1.72(m,2H),0.95(d,J=6.8Hz,6H)。¹³CNMR(100MHz,DMSO-d₆)146.8,130.0,128.9,128.8,125.7,123.4,89.6,82.1,70.7,40.6,38.3,28.1,28.0,22.3。ESIMSm/z232[M+1]⁺。

example 120

Preparation of 1- (3- (2-aminoethoxy) phenyl) hex-1-yn-3-ol

1- (3- (2-Aminoethoxy) phenyl) hex-1-yn-3-ol was prepared according to the method described for example 18.

Step 1: the bromide 19 was Sonogashira reacted with 4-methylpent-1-yn-3-ol to give 2,2, 2-trifluoro-N- (2- (3- (3-hydroxyhex-1-ynyl) phenoxy) ethyl) acetamide as a clear oil. Yield (3g, crude): the crude product was used directly for further deprotection reactions.

Step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (3-hydroxyhex-1-ynyl) phenoxy) ethyl) acetamide gave example 120 as a yellow oil. Yield (1.858g, 88%): ¹HNMR(400MHz,DMSO-d₆)7.23-7.29(m,1H),6.90-6.98(m,3H),4.42(t,J=6.4Hz,2H),3.92(t,J=4.8Hz,2H),2.86(bs,2H),1.56-1.68(m,2H),1.40-1.49(m,2H),0.91(t,J=7.6Hz,3H)。¹³CNMR(100MHz,DMSO-d₆)159.0,130.3,124.0,117.1,115.8,92.8,833,70.4,61.0,41.2,40.1,18.6,14.2。ESIMSm/z234[M+1]⁺。

Example 121

Preparation of 3-amino-1- (3- (4-methoxybut-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (4-methoxybut-1-ynyl) phenyl) propan-1-ol was prepared according to the method described for example 132.

Step 1: sonogashira reaction of 25 with 4-methoxybut-1-yne gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-methoxybut-1-ynyl) phenyl) propyl) acetamide as a dark yellow oil. Yield (0.51g, 51%). This compound can be carried forward to the next step without complete purification.

Step 2: 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-methoxybut-1-ynyl) phenyl) propyl) acetamide was deprotected as described in example 132 except that the reaction mixture was stirred at room temperature for 16 hours. Example 121 was obtained as a yellow oil. Yield (0.22g, 60%):¹HNMR(400MHz,DMSO-d₆)7.33(s,1H),7.28-7.29(m,2H),7.21-7.24(m,1H),4.66(t,J=6.4Hz,1H),3.50(t,J=6.4Hz,2H),3.28(s,3H),2.63-2.69(m,4H)1.65-1.77(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.5,129.4,128.5,128.2,125.4,122.6,87.7,81.1,70.4,70.1,57.8,40.6,40.1,19.8。ESIMSm/z234[M+1]⁺。

example 122

Preparation of 1- (3- (2-aminoethoxy) phenyl) -3-methylhexan-1-yn-3-ol

1- (3- (2-Aminoethoxy) phenyl) -3-methylhexan-1-yn-3-ol is prepared as described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with but-3-ynylbenzene to give 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-methylhexan-1-ynyl) phenoxy) ethyl) acetamide as a clear oil. Yield (2.0g, 71%): the crude product was used directly for further deprotection reactions.

Step 2: will 2Deprotection of 2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-methylhexan-1-ynyl) phenoxy) ethyl) acetamide gave example 122 as a pale yellow oil. Yield (0.700g, 35%):¹HNMR(400MHz,DMSO-d₆)7.23-7.28(m,1H),6.91-6.95(m,2H),6.87-6.89(m,1H),5.34(s,1H),3.91(t,J=6.0Hz,2H),2.85(t,J=5.6Hz,2H),1.45-1.70(m,5H),1.41(s,3H),0.89-0.94(m,3H)。¹³CNMR(100MHz,DMSO-d₆)158.6,129.8,123.7,123.6,116.6,115.2,81.4,70.3,66.6,45.9,40.9,29.9,17.7,14.3。ESIMSm/z248[M+1]⁺。

example 123

Preparation of (S) -1- ((3- (1-aminopropyl-2-yloxy) phenyl) ethynyl) cyclohexanol

(S) -1- ((3- (1-aminopropyl-2-yloxy) phenyl) ethynyl) cyclohexanol was prepared according to the procedure described in equation 20.

Reaction scheme 20

Step 1: diethyl azodicarboxylate (17.4g,100mmol) was slowly added to a solution of 3-iodophenol (18.5g,84mmol), alcohol (78) (14.73g,84mmol) and triphenylphosphine (26.2g,100mmol) in THF (200mL) at 0 deg.C under argon. The reaction was warmed to room temperature and stirred at room temperature for 2 hours, heated to 80 ℃ for 6 hours, and then concentrated under reduced pressure. The residue was triturated with diethyl ether and the resulting white solid was removed by filtration. The filtrate was concentrated under reduced pressure, and the residue was separated in ethyl acetate and 1N NaOH. The organics were combined, washed with brine, and concentrated under reduced pressure. The residue was purified by flash chromatography (5-20% etoac/hexanes gradient) on silica gel to give carbamate (79) as an impure yellow oil, which was taken to the next step without further purification. Yield (17.3g, 54%).

Step 2: HCl (12mL, 4.8M iPrOH solution, 56mmol) was added to a solution of carbamate (79) (10g,28mmol) in ethyl acetate (25 mL). After stirring for 1 hour, the reaction mixture was filtered and the solid was dried under reduced pressure to give the hydrochloride salt (80) as a white solid which was taken to the next step without purification or analysis. Yield (2.9g, 30%).

And step 3: the amine hydrochloride (80) was protected with ethyl trifluoroacetate as described for example 18 except that 1 equivalent of TEA was used and the reaction was carried out in dichloromethane to give the trifluoroacetamide (81) as a yellow oil. Yield (3.4g, quantitative).¹HNMR(400MHz,CDCl₃)7.29–7.33(m,1H),7.24–7.26(m,1H),6.99(t,J=8.0Hz,1H),6.83–6.87(m,1H),6.75(brs,1H),4.45–4.55(m,1H),3.52–3.53(m,1H),3.40–3.50(m,1H),1.29(d,J=6.4Hz,3H)。

And 4, step 4: a mixture of trifluoramide 81(500mg,1.34mmol), 1-ethynylcyclohexanol (250mg,2.01mmol), copper iodide (25mg,0.13mmol), tri-o-tolylphosphine (40mg,0.13mmol), TEA (0.279mL,2.01mL), and triphenylphosphine palladium dichloride (91mg,0.13mmol) in DMF (13mL) was degassed, placed under an argon atmosphere, and stirred at 90 ℃ overnight. The reaction mixture was filtered and the filtrate was separated in ethyl acetate/water. The organic layers were combined and washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (10-30% etoac-hexanes gradient) to afford alkyne 82 as a yellow glassy oil. Yield (0.322g, 65%). ¹HNMR(400MHz,CDCl₃)7.21(t,J=8.0Hz,1H),7.04–7.08(m,1H),6.94–6.96(m,1H),6.83–6.87(m,1H),6.81(brs,1H),4.48–4.57(m,1H),3.72–3.80(m,1H),3.39–3.49(m,1H),1.85–2.04(m,3H),1.50–1.80(m,8H),1.29(d,J=6.4Hz,3H)。

And 5: deprotection of alkyne 82 as described in example 1 affords example 123 as a yellow oil. Yield (0.200g, 85%).¹HNMR(400MHz,CDCl₃)7.18(t,J=8.0Hz,1H),6.96–7.02(m,2H),6.84–6.88(m,1H),4.30–4.38(m,1H),2.87(d,J=5.2Hz,2H),1.85–2.02(m,2H),1.50–1.80(m,11H),1.25(d,J=6.4Hz,3H)。ESIMSm/z274.3[m+H]⁺。

Example 124

Preparation of 1- (3- (2-aminoethoxy) phenyl) -4-methylpent-1-yn-3-ol

1- (3- (2-Aminoethoxy) phenyl) -4-methylpent-1-yn-3-ol is prepared as described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with 4-methylpent-1-yn-3-ol to give 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-4-methylpent-1-ynyl) phenoxy) ethyl) acetamide as a yellow oil. Yield (0.51g, 21%):¹HNMR(400MHz,CDCl₃)7.22-7.27(m,1H),7.08-7.12(d,J=7.6Hz,1H),6.98(s,1H),6.88(dd,1H,J=6.8Hz,1.6,1H),6.71(bs,1H),4.32(s,2H),4.09(t,J=5.2Hz,2H),3.77-3.82(m,2H),1.77-1.83(m,1H),0.94-0.99(m,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-4-methylpent-1-ynyl) phenoxy) ethyl) acetamide gave example 124 as a yellow oil. Yield (0.160g, 47%):¹HNMR(400MHz,DMSO-d₆)7.22-7.30(m,1H),6.90-7.00(m,3H),4.21(d,J=5.6Hz,1H),3.93(t,J=5.2Hz,2H),2.87(bs,2H),1.77-1.83(m,1H),0.94-0.99(m,6H)。¹³CNMR(100MHz,DMSO-d₆)158.5,129.8,123.7,116.7,115.2,91.0,83.6,69.9,66.3,40.7,34.3,18.3,17.7。ESIMSm/z234[M+1]⁺。

example 125

Preparation of 3-amino-1- (3- (phenylethynyl) phenyl) propan-1-ol

3-amino-1- (3- (phenylethynyl) phenyl) propan-1-ol is prepared as described in example 121.

And 4, step 4: sonogashira reaction of 25 with ethynylbenzene gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (2-phenylethynyl) phenyl) propyl) acetamide as a yellow oil. Yield (0.78g, 73%).

And 5: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (2-phenylethynyl) phenyl) propyl) acetamide gave example 125 as a white semi-solid. Yield (0.3g, 53%): ¹HNMR(400MHz,DMSO-d₆)7.51-7.53(m,2H),7.48(s,1H),7.34-7.42(m,6H),5.52(bs,2H),4.66(t,J=6.4Hz,1H),2.70(t,J=7.2Hz,2H),1.76(t,J=7.2Hz,2H)。¹³CNMR(100MHz,DMSO-d₆)146.9,131.8,130.2,129.3,129.0,126.6,122.7,122.4,89.9,89.5,70.5,40.6,38.0。ESIMSm/z252[M+1]⁺。

Example 126

Preparation of 5- (3- (3-amino-1-hydroxypropyl) phenyl) -N, N-dimethylpent-4-ynamide

5- (3- (3-amino-1-hydroxypropyl) phenyl) -N, N-dimethylpent-4-ynamide was prepared according to the procedure described for the preparation of example 121.

Step 1: sonogashira reaction of 4 with penta-4-ynoic acid dimethylamide gave 5- (3- (1-hydroxy-3- (2,2, 2-trifluoroacetamido) propyl) phenyl) -N, N-dimethylpent-4-ynamide as a dark yellow oil. Yield (0.33g, 48%). This compound had a trace amount of starting material and was used without further purification.

Step 2: deprotection of 5- (3- (1-hydroxy-3- (2,2, 2-trifluoroacetamido) propyl) phenyl) -N, N-dimethylpent-4-ynamide affords example 126 as a light yellow oil. Yield (0.147g, 60%):¹HNMR(400MHz,DMSO-d₆)7.64(bs,2H),7.34(s,1H),7.24-7.32(m,3H),5.59(bs,1H),4.66(t,J=5.2Hz,1H),2.97(s,3H),2.83(s,3H),2.80(m,2H),2.60(s,4H),1.75-1.88(m,2H)。¹³CNMR(100MHz,DMSO-d₆)170.2,145.6,129.7,128.5,125.2,123.0,90.2,80.3,69.3,40.1,36.4,36.3,34.9,31.7,14.8。ESIMSm/z275[M+1]⁺。

example 127

Preparation of 3- (3- (cyclopropylethynyl) phenyl) propan-1-amine

3- (3- (cyclopropylethynyl) phenyl) propan-1-amine is prepared according to the procedure described in equation 21.

Reaction formula 21

Step 1: to LiAlH at room temperature₄To a suspension of (0.5g,1.3mmol) in diethyl ether (25mL) was added 3-bromocinnamic acid (83,1.0g,4mmol) in portions. The resulting suspension was stirred for 3 hours. The reaction was quenched by the sequential addition of 15% aqueous NaOH (1mL) and water. The resulting white suspension was filtered through a pad of celite. The filter cake was washed with ether and then ethyl acetate. The filtrate was concentrated to give crude product 8 as a yellow oil. Yield (0.7g, 74%). ¹HNMR(400MHz,CDCl₃)7.10-7.35(m,4H),3.65(t,J=6.5Hz,2H),2.68(t,J=7.8Hz,2H),1.82-1.91(m,2H)。

Step 2: to a stirred solution of alcohol 8(1.0g,4.6mmol) and triethylamine (1.0mL,99mmol) in DCM (15mL) was added MsCl (0.7mL,66mmol) at 0 deg.C for 5 min. The resulting mixture was stirred at 0 ℃ for 30 minutes, warmed to room temperature and stirred for 30 minutes during which time the conversion was complete. The mixture was poured into water and extracted with ethyl acetate. The combined organic layers were washed with water and brine, and anhydrous Na₂SO₄And (5) drying. It was filtered and concentrated to give mesylate (mesylate)84 as a yellow oil. (1.0g, 73%). This crude product was immediately used in the next conversion process.

And step 3: to a solution of crude mesylate 84(1.0g,3.4mmol) in DMF (8mL) was added NaN₃(0.44g,6.8 mmol). The resulting mixture was stirred at 80 ℃ for 1 hour. It was cooled, poured into water, and extracted with ethyl acetate. The combined organics were washed with water and brine, washed with anhydrous Na₂SO₄And (5) drying. Filtered and then concentrated under reduced pressure to give 85 as a colorless oil. Yield (0.7g, 85%).¹HNMR(400MHz,CDCl₃)7.32-7.38(m,2H),7.09-7.20(m,2H),3.29(t,J=6.4Hz,2H),2.68(t,J=7.2Hz,2H),1.84-1.94(m,2H)。

And 4, step 4: to a solution of azide 85(15g,62.7mmol) in a mixture of THF (133mL) and water (13mL) was added Ph at room temperature₃P (16g,62.7 mmol). The mixture was stirred for 48 hours during which time the conversion process was complete. The solvent was removed under reduced pressure and the resulting residue was used in the next step.

And 5: amine 86 was dissolved in 1, 4-dioxane (300mL) and water (180 mL). Adding K in sequence₂CO₃(17.2g,120mmol)、(Boc)₂O (14mL,60mmol) and the mixture was stirred for 2 hours. After removal of 1, 4-dioxane under reduced pressure, the aqueous phase was extracted with ethyl acetate. The organic phase was washed with water and brine, over anhydrous Na₂SO₄And (5) drying. It was concentrated to give the crude product as a yellow oil. Purification by flash chromatography (0-9% etoac-hexanes gradient) gave 87 as a pale yellow oil. Yield (15g, 79%).¹HNMR(400MHz,CDCl₃)7.30-7.35(m,2H),7.09-7.2(m,2H),4.53(bs,1H),3.14(m,2H),2.61(t,J=7.6Hz,2H),1.74-1.84(m,2H),1.45(s,9H)。

Step 6: to a degassed solution of bromide 87(1.0g,3.1mmol) and cyclopropylacetylene (2.9mL,3.4mmol,70% in toluene) in diisopropylamine (4mL) was added PdCl₂(PPh₃)₂(0.120g,0.17mmol), tri-o-tolylphosphine (0.048g,0.16mmol), and CuI (0.026g,0.16 mmol). The resulting mixture was degassed and stirred overnight at 90 ℃ under nitrogen. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was separated with water and ethyl acetate. Applying anhydrous Na to the organic layer₂SO₄Dried, filtered, and concentrated under reduced pressure. Purification by flash chromatography (10-40% etoac-hexanes gradient) afforded tert-butyl [3- (3-cyclopropylethynylphenyl) propyl carbamate (88). Yield (0.756g, 79%). This alkyne was used for deprotection without further purification.

And 7: alkyne 88 was dissolved in THF (4.0mL) and HCl in dioxane (5mL,4M) was added. The mixture was stirred at room temperature for 18 hours. Concentration under reduced pressure followed by trituration with hexanes afforded example 127 hydrochloride as a yellow semi-solid. Yield (0.2g, 85%):¹HNMR(400MHz,DMSO-d₆.)7.22-7.27(m,1H),7.13-7.19(m,3H),2.74(t,J=7.2Hz,2H),2.55(t,J=7.6Hz,2H),1.74-1.82(m,2H),1.44-1.51(m,1H),0.84-0.89(m,2H),0.64-0.68(m,2H)。¹³CNMR(100MHz,DMSO-d₆):141.2,131.1,128.9,127.9,123.2,93.6,75.6,8.1,31.4,28.4,8.3,-0.3。ESIMSm/z200[M+1]⁺。

example 128

Preparation of 2- (3- (4-methoxybut-1-ynyl) phenoxy) ethylamine

2- (3- (4-Methoxybut-1-ynyl) phenoxy) ethylamine was prepared as described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with 4-methoxybut-1-yne to give 2,2, 2-trifluoro-N- (2- (3- (4-methoxybut-1-ynyl) phenoxy) ethyl) acetamide as a yellow oil. Yield (0.45g, 45%): this material was used directly for deprotection reactions.

Step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (4-methoxybut-1-ynyl) phenoxy) ethyl) acetamide gave example 128 as a yellow oil. Yield (0.120g, 50%):¹HNMR(400MHz,DMSO-d₆)7.21-7.26(m,1H),6.89-6.96(m,3H),3.90(t,J=5.6Hz,2H),3.49(t,J=6.4Hz,2H),3.29(s,3H),2.84(t,J=5.6Hz,2H),2.65(t,J=6.4Hz,2H)。¹³CNMR(100MHz,DMSO-d₆)158.6,129.7,124.1,123.6,116.7,115.0,87.9,80.9,70.3,70.0,57.9,40.9,19.9。ESIMSm/z220[M+1]⁺。

example 129

Preparation of 1- (2- (3- (2-aminoethoxy) phenyl) ethynyl) cyclooctanol

1- (2- (3- (2-aminoethoxy) phenyl) ethynyl) cyclooctanol was prepared according to the method described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with 1-ethynylcyclooctanol to give 2,2, 2-trifluoro-N- (2- (3- (2- (1-hydroxycyclooctyl) ethynyl) phenoxy) ethyl) acetamide as a clear oil. Yield (1.3g, 72%): ¹HNMR(400MHz,DMSO-d₆)7.22(d,J=8.0Hz,1H),7.07(d,J=7.6Hz,1H),6.91-6.96(m,2H),4.09-4.13(m,2H),2.00-2.06(m,6H),1.48-1.72(m,11H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (2- (1-hydroxycyclooctyl) ethynyl) phenoxy) ethyl) acetamide to give a yellow oily solidExample 129. Yield (1.858g, 88%):¹HNMR(400MHz,DMSO-d₆)7.24-7.29(m,1H),6.91-6.96(m,2H),6.89(s,1H),3.94(t,J=5.6Hz,2H),2.88(t,J=5.6Hz,2H),1.80-1.92(m,5H),1.50-1.60(m,7H),1.42-1.44(m,3H)。¹³CNMR(100MHz,DMSO-d₆)158.5,129.8,123.8,123.6,116.7,115.1,95.7,81.6,69.7,40.6,37.7,27.6,24.1,21.7。ESIMSm/z234[M+1]⁺。

example 130

Preparation of 5- (3- (3-amino-1-hydroxypropyl) phenyl) pent-4-yn-1-ol

5- (3- (3-amino-1-hydroxypropyl) phenyl) pent-4-yn-1-ol was prepared according to the method described for example 132.

Step 1: sonogashira reaction of 25 with pent-4-yn-1-ol gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (5-hydroxypent-1-ynyl) phenyl) propyl) acetamide as a brown oil. Yield (1.46g, 69%):¹HNMR(400MHz,CDCl₃)7.38(s,1H),7.22-7.34(m,3H),4.86(d,J=8.0Hz,1H),3.83(t,J=5.2Hz,2H),3.65-3.69(m,1H),3.38-3.42(m,1H),2.56(t,J=7.2Hz,2H),2.38(bs,1H)1.93-1.99(m,2H),1.83-1.88(m,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (5-hydroxypent-1-ynyl) phenyl) propyl) acetamide gave example 130 as a yellow oil. Yield (0.64g, 65%):¹HNMR(400MHz,DMSO-d₆)7.32(s,1H),7.25-7.28(m,2H),7.20-7.22(m,1H)4.66(t,J=6.4Hz,1H),4.55(bs,1H),3.52(t,J=6.4Hz,2H),2.57-2.66(m,2H),2.44(t,J=7.2Hz,2H),1.60-1.70(m,4H)。¹³CNMR(100MHz,DMSO-d₆)147.2,129.8,129.0,128.7,125.7,123.3,90.6,81.1,71.1,59.9,41.8,38.9,32.0,15.7。ESIMSm/z234[M+1]⁺。

example 131

Preparation of 3-amino-1- (3- (cyclopropylethynyl) phenyl) propan-1-ol

3-amino-1- (3- (2-cyclopropylethynyl) phenyl) propan-1-ol was prepared as described for example 132, except that the deprotection was carried out at room temperature for 16 hours.

Step 1: sonogashira reaction of 25 with ethynylcyclopropane gave N- (3- (3- (2-cyclopropylethynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a dark brown oil. Yield (0.8g, 83%). It was used in the next conversion without further purification.

Step 2: deprotection of N- (3- (3- (2-cyclopropylethynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide affords example 131 as a yellow oil. Yield (0.13g, 24%):¹HNMR(400MHz,DMSO-d₆)7.23-7.33(m,4H),4.64(t,J=6.4Hz,1H),2.71(t,J=7.2Hz,2H),1.65-1.73(m,2H),1.50–1.56(m,1H),0.86-0.91(m,2H),0.69-0.73(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.2,129.5,128.5,128.3,125.1,122.8,93.5,75.7,70.1,40.1,37.6,8.34,-0.3。ESIMSm/z216[M+1]⁺。

example 132

1- (3- (3-amino-1-hydroxypropyl) phenyl) hex-1-yn-3-ol

1- (3- (3-amino-1-hydroxypropyl) phenyl) hex-1-yn-3-ol was prepared according to the method described for example 107.

Step 1: sonogashira reaction of 25(3g,9.2mmol) with hex-1-yn-3-ol gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxyhex-1-ynyl) phenyl) propyl) acetamide as a yellow oil. Yield (2.31g, 73%):¹HNMR(400MHz,CDCl₃)7.42(s,1H),7.26-7.38(m,3H),4.86(m,1H),4.61(dd,J=2.0,5.6Hz,1H),3.67-3.71(m,1H),3.37-3.46(m,1H),2.38(d,J=2.0Hz,1H),1.95-1.99(m,2H),1.75-1.88(m,2H),1.53-1.57(m,2H),0.97(t,J=7.2Hz,3H)。

step 2: a mixture of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxyhex-1-ynyl) phenyl) propyl) acetamide, potassium carbonate (0.438g,3.26mmol) and water (2mL) in 2-PrOH (10mL) was heated at reflux overnight. The reaction mixture was concentrated to dryness under reduced pressure. Purification by flash chromatography eluting with 10% (9.5:0.5 methanol-ammonia) -DCM gave example 132 as a yellow oil. Yield (0.42g, 49%):¹HNMR(400MHz,DMSO-d₆)7.23-7.36(m,4H),6.90(bs,2H),5.44(d,J=4.8Hz,1H),4.67(t,J=7.6Hz,1H),4.44(d,J=5.2Hz,1H),2.82(t,J=7.6Hz,2H),1.79-1.83(m,2H),1.60-1.64(m,2H),1.41-1.47(m,2H),0.93(t,J=7.2Hz,3H)。¹³CNMR(100MHz,DMSO-d₆)145.8,129.7,128.5,128.4,125.6,122.3,92.4,83.0,69.5,60.5,37.0,36.6,18.2,13.7。ESIMSm/z248[M+1]⁺。

example 133

Preparation of 3- (3- (4-methylpent-1-ynyl) phenyl) propan-1-amine

3- (3- (4-methylpent-1-ynyl) phenyl) propan-1-amine is prepared as described in example 127.

Step 1: sonogashira coupling of bromide 87(0.5g,1.5mmol) with 4-methyl-1-pentyne (0.2mL,2.4mmol) gave tert-butyl 3- (4-methylpent-1-ynyl) phenylcarbamate. The yield (0.35g, 69%)。¹HNMR(400MHz,CDCl₃)7.16-7.28(m,3H),7.07(d,J=7.2Hz,1H),4.50(bs,1H),3.12-3.15(m,2H),2.60(d,J=7.6Hz,2H),2.29(d,J=6.8Hz,2H),1.84-1.94(m,2H),1.74-1.82(m,1H),1.44(s,9H),1.04(d,J=6.8Hz,6H)。

Step 2: tert-butyl 3- (4-methylpent-1-ynyl) phenylcarbamate was deprotected with HCl/dioxane in THF to give example 133 hydrochloride as a light yellow solid. Yield (0.2g, 64%):¹HNMR(400MHz,DMSO-d₆)7.23-7.29(m,1H),7.13-7.21(m,3H),2.76(t,J=7.6Hz,2H),2.56(t,J=7.6Hz,2H),2.25(d,J=6.4Hz,2H),1.75-1.83(m,3H),0.94(d,J=6.8Hz,6H)。¹³CNMR(100MHz,DMSO-d₆):141.2,131.0,128.9,128.6,127.9,123.3,38.1,31.4,28.4,27.6,27.5,21.7。ESIMSm/z216[M+1]⁺。

example 134

Preparation of 5- (3- (2-aminoethoxy) phenyl) -N-methylpent-4-yneamide

5- (3- (2-Aminoethoxy) phenyl) -N-methylpent-4-ynoamide was prepared according to the method described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with pent-4-ynoic acid methylamide to give N-methyl-5- (3- (2- (2,2, 2-trifluoroacetamido) ethoxy) phenyl) pent-4-ynylamide as a clear oil. Yield (1.4g, crude): the crude product was used in the next step without further purification.

Step 2: n-methyl-5- (3- (2- (2,2, 2-trifluoroacetamido) ethoxy) phenyl) pent-4-ynylamide was deprotected to give example 134 as a brown solid. Yield (0.110g, 10%):¹HNMR(400MHz,DMSO-d₆)7.27-7.31(m,1H),6.95-7.00(m,3H),4.14-4.17(m,2H),3.18(t,J=5.2Hz,2H),2.58-2.62(m,2H),2.58(s,3H),2.32(t,J=7.2Hz,2H)。¹³CNMR(100MHz,DMSO-d₆)170.6,157.7,129.8,124.3,124.2,117.0,115.1,90.0,80.2,64.4,38.2,34.2,25.4,15.2。ESIMSm/z266[M+1]⁺。

example 135

Preparation of 5- (3- (2-aminoethoxy) phenyl) -N, N-dimethylpent-4-yneamide

5- (3- (2-Aminoethoxy) phenyl) -N, N-dimethylpent-4-ynylamide prepared according to the procedure described in the examples.

Step 1: the bromide 19 was Sonogashira reacted with pent-4-ynoic acid N, N-dimethylamide to give N, N-dimethyl-5- (3- (2- (2,2, 2-trifluoroacetamido) ethoxy) phenyl) pent-4-ynylamide as a brown oil. Yield (0.9g, 50%): this crude material was used in the next step without further purification.

Step 2: n, N-dimethyl-5- (3- (2- (2,2, 2-trifluoroacetamido) ethoxy) phenyl) pent-4-ynylamide was deprotected to give example 135 as a brown oil. Yield (0.14g, 55%):¹HNMR(400MHz,DMSO-d₆)7.22-7.28(m,1H),6.90-6.97(m,3H),4.0(t,J=5.6Hz,2H),2.94-3.0(m,2H),2.83(s,6H),2.57-2.62(m,4H)。¹³CNMR(100MHz,DMSO-d₆)170.2,158.2,158.0,157.7,129.8,124.3,123.9,116.8,115.0,90.3,80.2,36.6,34.9,31.6,14.8。ESIMSm/z261[M+1]⁺。

example 136

Preparation of 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclooctanol

1- (2- (3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclooctanol was prepared according to the method described for example 131.

Step 1: sonogashira reaction of 25 with 1-ethynylcyclooctanol gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (2- (1-hydroxycyclooctyl) ethynyl) phenyl) propyl) acetamide as a yellow oil. Yield (0.54g, 44%). This compound was used in the next step without further purification.

Step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (2- (1-hydroxycyclooctyl) ethynyl) phenyl) propyl) acetamide afforded example 136 as a white solid. Yield (0.26g, 64%):¹HNMR(400MHz,DMSO-d₆)7.20-7.29(m,4H),5.23(s,1H),4.66(t,J=6.4Hz,1H),2.55-2.62(m,2H),1.79–1.90(m,4H),1.62-1.66(m,2H),1.55-1.60(m,8H),1.40-1.42(m,2H)。¹³CNMR(100MHz,DMSO-d₆)147.0,129.2,128.5,128.2,125.6,122.3,95.4,81.9,70.8,69.7,42.4,40.1,39.9,37.7,27.5,24.0,21.7。ESIMSm/z302[M+1]⁺。

example 137

Preparation of 5- (3- (2-aminoethoxy) phenyl) -pent-4-ynylamide

5- (3- (2-Aminoethoxy) phenyl) -pent-4-ynylamide prepared according to the procedure described in example 18.

Step 1: the bromide 6 was Sonogashira reacted with pent-4-ynoic acid amide to give 5- (3- (2- (2,2, 2-trifluoroacetamido) ethoxy) phenyl) pent-4-ynoic acid amide as a clear oil. Yield (0.8g, 50%): this compound was used in the next step without further purification.

Step 2: deprotection of 5- (3- (2- (2,2, 2-trifluoroacetylamino) ethoxy) phenyl) pent-4-ynylamide afforded example 137 as a pale yellow oil. Yield (0.093g, 16%):¹HNMR(400MHz,DMSO-d₆)7.21-7.26(m,1H),6.88-6.94(m,3H),3.92(t,J=5.6Hz,2H),2.88(t,J=5.6Hz,2H),2.59(t,J=7.2Hz,2H),2.34(t,J=7.2Hz,2H)。¹³CNMR(100MHz,DMSO-d₆)172.4,158.5,129.7,124.2,123.7,116.8,114.9,89.9,80.3,69.6,40.6,34.1,15.0。ESIMSm/z233[M+1]⁺。

example 138

5- (3- (3-amino-1-hydroxypropyl) phenyl) -N-methylpent-4-ynylamide octanol

5- (3- (3-amino-1-hydroxypropyl) phenyl) -N-methylpent-4-ynoamide is prepared according to the method described in scheme 22.

Reaction formula 22

Step 1: to a solution of 24(17g,74mmol) in DCM (250mL) was added (Boc)₂O (21.5mL,89mmol) and triethylamine (15.5mL,111 mmol). The reaction mixture was stirred at room temperature for 15 h, then diluted with DCM (250mL) and saturated NaHCO₃And (4) washing the solution. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. Purification by flash chromatography (0-30% etoac-hexanes gradient) afforded 89 as a yellow oil. Yield (15.2g, 61%):¹HNMR(400MHz,DMSO-d₆)7.53(s,1H),7.39(d,J=7.6Hz,1H),7.29(d,J=7.6Hz,1H),7.20(t,J=7.2Hz,1H),4.85(bs,1H),4.69-4.71(m,1H),3.60(bs,1H),3.53-3.55(m,1H),3.11-3.18(m,1H),1.76-1.84(m,2H),1.47(s,9H)。

step 2: sonogashira reaction of 89 with pent-4-ynoic acid methylamide was carried out to give tert-butyl 3-hydroxy-3- (3- (5- (methylamino) -5-oxopent-1-ynyl) phenyl) propylcarbamate as a dark brown oil. Yield (0.27g, 36%):¹HNMR(400MHz,DMSO-d₆)7.39(s,1H),7.26-7.30(m,3H),5.64(bs,1H),4.85(bs,1H),4.69-4.71(m,1H),3.46-3.51(m,1H),3.35(bs,1H),3.11-3.18(m,1H),2.85(d,J=4.8Hz,3H),2.73(t,J=6.4Hz,2H),2.46(t,J=6.4Hz,2H),1.55-1.84(m,2H),1.46(s,9H)。

and step 3: to a solution of 3-hydroxy-3- (3- (5- (methylamino) -5-oxopent-1-ynyl) phenyl) propylcarbamic acid tert-butyl ester (0.7g,2.1mmol) in methanol-THF (1:1,10mL) was added HCl in dioxane (0.7mL,4M), and the resulting mixture was stirred at room temperature for 24 hours. The mixture was evaporated to dryness under reduced pressure. Purification by flash chromatography (0-15% gradient methanol-DCM) afforded example 14 hydrochloride as a yellow solid. Yield (0.17g, 76%): ¹HNMR(400MHz,DMSO-d₆)7.27-7.33(m,3H),7.23-7.27(m,1H),4.64-4.66(m,1H),2.75-2.82(m,2H),2.60-2.62(m,2H),2.58(s,3H),2.33-2.35(m,2H),1.78-1.90(m,2H)。¹³CNMR(100MHz,DMSO-d₆)171.1,146.1,130.3,128.9,128.8,125.7,123.4,90.3,81.0,69.7,36.9,36.8,34.8,25.9,15.6。ESIMSm/z261[M+1]⁺。

Example 139

3-amino-1- (3- ((2, 6-dichlorophenyl) ethynyl) phenyl) propan-1-ol

3-amino-1- (3- ((2, 6-dichlorophenyl) ethynyl) phenyl) propan-1-ol was prepared according to the procedure described for example 132.

Step 1: sonogashira reaction of 25 with 1, 3-dichloro-2-ethynylbenzene gave N- (3- (3- (2- (2, 6-dichlorophenyl) ethynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a dark brown oil. Yield (0.32g, 37%). This compound cannot be completely isolated from the starting bromide and is used directly in the next step.

Step 2: n- (3- (3- (2- (2, 6-dichlorophenyl) ethynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide was deprotected to give example 139 as a yellow solid. Yield (0.149g, 60%):¹HNMR(400MHz,DMSO-d₆)7.69(bs,2H),7.63(d,J=8.4Hz,2H),7.51(s,1H),7.43-7.53(m,4H),5.71(bs,1H),4.66(m,1H),2.82-2.90(m,2H),1.82-1.95(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.2,136.0,130.8,128.9,128.5,128.2,127.0,121.8,121.2,118.8,115.8,99.6,83.0,69.3,36.5,36.4。ESIMSm/z320[M+1]⁺。

example 140

5- (3- (3-amino-1-hydroxypropyl) phenyl) -N-methylpent-4-ynylamide octanol

5- (3- (3-amino-1-hydroxypropyl) phenyl) -N-methylpent-4-ynoamide was prepared according to the method described in example 138.

Step 1: sonogashira reaction of 24 with 1-ethynylcyclobutanol gave tert-butyl 3-hydroxy-3- (3- (2- (1-hydroxycyclobutyl) ethynyl) phenyl) propylcarbamate as a yellow oil. Yield (1.5g, 70%):¹HNMR(400MHz,DMSO-d₆)7.45(s,1H),7.27-7.34(m,3H),4.85(bs,1H),4.71(m,1H),3.48-3.51(m,2H),3.14-3.16(m,1H),2.50-2.52(m,2H),2.30-2.49(m,3H),1.80-1.90(m,4H),1.45(s,9H)。

step 2: deprotection of tert-butyl 3-hydroxy-3- (3- (2- (1-hydroxycyclobutyl) ethynyl) phenyl) propylcarbamate afforded example 140 hydrochloride as a light yellow semi-solid. Yield (0.161g, 30%): ¹HNMR(400MHz,DMSO-d₆)7.28-7.36(m,4H),4.64-4.67(m,1H),2.77-2.86(m,2H),2.30-2.39(m,2H),2.16-2.24(m,2H),1.70-1.90(m,4H)。¹³CNMR(100MHz,DMSO-d₆)145.8,129.7,128.5,128.4,125.6,122.3,94.6,81.6,69.1,66.2,40.1,36.5,36.2,12.8。ESIMSm/z246[M+1]⁺。

Example 141

Preparation of 4- ((3- (2-aminoethyl) phenyl) ethynyl) hept-4-ol

4- ((3- (2-aminoethyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure described for example 18.

Step 1: 3-Bromophenylethylamine was condensed with ethyl trifluoroacetate to give N- (3-bromophenylethyl) -2,2, 2-trifluoroacetamide as a colorless oil. Yield (3.30g, quantitative).¹HNMR(400MHz,CDCl₃)7.40(ddd,J=7.8,1.8,1.0Hz,1H),7.53(t,J=1.8Hz,1H),7.21(t,J=7.6Hz,1H),7.10-7.13(m,1H),6.32(brs,1H),3.61(q,J=6.7Hz,2H),2.87(t,J=7.2Hz,2H)。

Step 2: with the exception that the reaction was carried out for 17 hours, N- (3-bromophenylethyl) -2,2, 2-trifluoroacetamide was coupled with alkynol 20 as described in example 18 to give 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenethyl) acetamide as a brown oil after purification by flash chromatography (10% to 50% gradient of ethyl acetate in hexane). Yield (0.975g, 66%).¹HNMR(400MHz,CDCl₃)7.32(dt,J=7.8,1.2Hz,1H),7.24-7.29(m,2H),7.10-7.15(m,1H),6.27(br.s,1H),3.61(q,J=6.7Hz,2H),2.86(t,J=7.0Hz,2H),1.95(s,1H),1.66-1.74(m,4H),1.52-1.64(m,4H),0.99(t,J=7.2Hz,6H)。

And step 3: 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenethyl) acetamide was reacted as described in example 1 except that the reaction was stirred at 40 ℃ for 18 hoursAnd (4) deprotection. Purification by flash chromatography (75-100% etoac-hexanes gradient 20% 7N ammonia/methanol) afforded example 141 as a colorless oil. Yield (0.385g, 54%).¹HNMR(400MHz,DMSO-d₆)7.21-7.26(m,1H),7.14-7.29(m,3H),5.12(s,1H),2.72(t,J=7.0Hz,2H),2.59(t,J=7.2Hz,2H),1.40-1.60(m,8H),0.89(t,J=7.0Hz,6H)；¹³CNMR(100MHz,DMSO-d₆)141.7,132.2,129.4,129.2,123.2,94.6,83.4,70.3,44.9,44.2,40.1,18.0,15.0；ESIMSm/z260.4[M+H]⁺;RP-HPLC100.0%(AUC,220nm)。

Example 142

Preparation of 3-amino-1- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propan-1-one oxime

3-amino-1- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propan-1-one oxime was prepared as described for examples 18 and 11.

Step 1: with the exception that the reaction was carried out at 80 ℃ for 3 hours, N- (3- (3-bromophenyl) -3-oxopropyl) -2,2, 2-trifluoroacetamide (63) was coupled with alkynol 20 according to the method described in example 18 to give 3-amino-1- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propan-1-one as a dark amber oil after purification by flash chromatography (ethyl acetate in 20% hexane). Yield (28.1g, 99.6%).¹HNMR(400MHz,DMSO-d₆)9.40(br.t,1H),7.84-7.92(m,2H),7.58-7.63(m,1H),7.50(t,J=7.6Hz,1H),5.19(s,1H),3.51(q,J=5.7Hz,2H),3.30(t,J=6.7Hz,2H),1.40-1.63(m,8H),0.90(t,J=7.2Hz,6H)。

Step 2: to a solution of 3-amino-1- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) propan-1-one (0.236g,0.62mmol) and hydroxylamine hydrochloride (0.109g,1.56mmol) in ethanol (anhydrous, 10mL) was added diisopropylethylamine (0.3mL,1.72mmol), and the reaction mixture was stirred at room temperature for 3 days. The residue was concentrated under reduced pressure and,flash chromatography (10-100% etoac in hexanes gradient) afforded 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3- (hydroxyimino) propyl) acetamide as a colorless oil. Yield (0.225g, 91%).¹HNMR(400MHz,DMSO-d₆)11.5(s,1H),9.50(t,J=5.5Hz,1H),7.55-7.62(m,2H),7.30-7.40(m,2H),5.14(s,1H),3.34(q,J=6.8Hz,2H),2.92(t,J=7.8Hz,2H),1.39-1.62(m,8H),0.89(t,J=7.2Hz,6H)。

And step 3: 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3- (hydroxyimino) propyl) acetamide was deprotected as described in example 1 except that the reaction was stirred at 40 ℃ for 18 hours. Purification by flash chromatography (50% to 100% gradient ethyl acetate-20% 7N ammonia in hexanes/methanol) afforded example 142 as a colorless oil. Yield (0.056g, 33%). ¹HNMR(400MHz,DMSO-d₆)7.59-7.62(m,2H),7.30-7.37(m,2H),5.15(s,1H),2.74-2.80(m,2H),2.60-2.66(m,2H),1.39-1.63(m,8H),0.89(t,J=7.4Hz,6H)；¹³CNMR(100MHz,DMSO-d₆)155.4,137.4,131.8,129.5,129.0,126.5,123.5,95.1,83.1,70.3,44.9,39.5,30.8,18.0,15.0；ESIMSm/z260.4[M+H]⁺；RP-HPLC100.0%(AUC,220nm)。

Example 143

Preparation of 2- ((3- (3-aminopropyl) phenyl) ethynyl) cyclohexanol

2- ((3- (3-aminopropyl) phenyl) ethynyl) cyclohexanol was prepared as described in example 1:

step 1: sonogashira coupling of bromide 3 with 2-ethynylcyclohexanol followed by purification by flash chromatography (5-50% etoac-hexanes gradient) gave 2,2, 2-trifluoro-N- (3- (3- ((2-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide as a yellow oil. The yield (1.2g,43%)：¹HNMR(400MHz,CDCl₃)7.18–7.28(m,3H),7.06–7.30(m,3H),6.33(brs,1H),3.48–3.57(m,1H),3.36(appq,J=6.8Hz,2H),2.63(t,J=7.2Hz,2H),2.38–2.46(m,1H),2.32(brs,1H),2.02–2.10(m,2H),1.91(quint,J=7.2Hz,2H),1.74–1.82(m,1H),1.66–1.74(m,1H),1.40–1.52(m,1H),1.16–1.40(m,4H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3- ((2-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide followed by flash chromatography (10% (7N ammonia/methanol)/dichloromethane) gave example 143 as an orange oil. Yield (0.606g, 69%):¹HNMR(400MHz,CDCl₃)7.16–7.26(m,3H),7.08–7.12(m,1H),3.48–3.56(m,1H),2.71(t,J=7.2Hz,2H),2.61(t,J=7.2Hz,2H),2.38–2.46(m,1H),2.01–2.10(m,2H),1.64–1.82(m,7H),1.40–1.52(m,1H),1.16–1.40(m,3H)。

example 144

Preparation of 2- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclohexanol

2- ((3- (2-aminoethoxy) phenyl) ethynyl) cyclohexanol was prepared as described for example 18:

step 1: sonogashira coupling of bromide 19 with 2-ethynylcyclohexanol was followed by purification by flash column chromatography (5-50% etoac-hexanes gradient) to give 2,2, 2-trifluoro-N- (2- (3- ((2-hydroxycyclohexyl) ethynyl) phenoxy) ethyl) acetamide as a yellow oil. Yield (0.88g, 31%): ¹HNMR(400MHz,CDCl₃)7.18(t,J=8.0Hz,1H),7.01–7.04(m,1H),6.90–6.98(brs,1H),6.91–6.92(m,1H),6.79–6.83(m,1H),4.05–4.07(m,2H),3.74(appq,J=5.2Hz,2H),3.48–3.56(m,1H),2.35–2.46(m,2H),2.00–2.08(m,1H),1.64–1.80(m,2H),1.40–1.52(m,1H),1.14–1.40(m,4H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- ((2-hydroxycyclohexyl) ethynyl) phenoxy) ethyl) acetamide followed by purification by flash chromatography (10% (7N ammonia/methanol)/dichloromethane) afforded example 144 as a white solid. Yield (0.29g, 45%):¹HNMR(400MHz,CDCl₃)7.18(t,J=7.6Hz,1H),6.98–7.03(m,1H),6.93–6.95(m,1H),6.83–6.86(m,1H),3.96(t,J=5.2Hz,2H),3.49–3.57(m,1H),3.08(brs,2H),2.38–2.46(m,1H),2.01–2.10(m,2H),1.55–2.00(brs,1H),1.74–1.82(m,2H),1.65–1.74(m,2H),1.40–1.52(m,1H),1.16–1.40(m,3H)。

example 145

Preparation of 2- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclohexanol

2- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclohexanol was prepared as described for example 19:

step 1: sonogashira coupling of bromide 25 with 2-ethynylcyclohexanol was followed by flash column chromatography (5-50% etoac-hexanes gradient) to give 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((2-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide as a yellow oil. Yield (1.9g, 63%):¹HNMR(400MHz,CDCl₃)7.39–7.41(m,1H),7.23–7.36(m,4H),4.84(q,J=4.0Hz,1H),3.62–3.72(m,1H),3.50–3.57(m,1H),3.34–3.44(m,1H),2.38–2.46(m1H),2.18(brs,2H),1.90–2.10(m,4H),1.66–1.84(m,2H),1.40–1.53(m,1H),1.16–1.40(m,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((2-hydroxycyclohexyl) ethynyl) phenyl) propyl) acetamide followed by flash chromatography (10% (7N ammonia/methanol)/dichloromethane) gave example 145 as a light yellow glassy solid. Yield (0.402g, 29%):¹HNMR(400MHz,CDCl₃)7.44–7.46(m,1H),7.21–7.31(m,3H),4.92(dd,J=8.8,3.2Hz,1H),3.47–3.56(m,1H),3.05–3.12(m,1H),3.01(brs,4H),2.90–2.99(m,1H),2.37–2.44(m,1H),2.00–2.09(m,2H),1.81–1.90(m,1H),1.64–1.81(m,3H),1.40–1.52(m,1H),1.14–1.40(m,3H)。

example 146

Preparation of 1- (2- (3- (2-aminoethoxy) phenyl) ethynyl) cyclobutanol

1- (2- (3- (2-aminoethoxy) phenyl) ethynyl) cyclobutanol was prepared according to the procedure described for example 18.

And 7: the bromide 19 was Sonogashira reacted with 1-ethynylcyclobutanol to give 2,2, 2-trifluoro-N- (2- (3- (2- (1-hydroxycyclobutyl) ethynyl) phenoxy) ethyl) acetamide as a brown oil. Yield (0.85g, 39%):¹HNMR(400MHz,DMSO-d₆)7.27-7.30(m,1H),6.92-7.03(m,3H),4.09-4.13(m,4H),3.54-3.58(m,2H),2.33-2.37(m,2H),2.16-2.24(m,2H),1.74-1.81(m,2H)。

and 8: deprotection of 2,2, 2-trifluoro-N- (2- (3- (2- (1-hydroxycyclobutyl) ethynyl) phenoxy) ethyl) acetamide gave example 146 as a brown oil. Yield (0.09g, 31%):¹HNMR(400MHz,DMSO-d₆)7.25-7.29(m,1H),6.98(d,J=8.0Hz,1H),6.93-6.96(m,2H),3.95(t,J=5.4Hz,2H),2.88(t,J=5.4Hz,2H),2.33-2.35(m,2H),2.16-2.24(m,2H),1.71-1.78(m,2H)。¹³CNMR(100MHz,DMSO-d₆)158.5,129.8,123.7,123.6,116.7,115.4,94.6,81.5,69.5,66.6,38.6,12.8。ESIMSm/z232[M+1]⁺。

example 147

Preparation of 1- (3- (3-amino-1-hydroxypropyl) phenyl) -4-methylpent-1-yn-3-ol

1- (3- (3-amino-1-hydroxypropyl) phenyl) -4-methylpent-1-yn-3-ol is prepared by the method described in example 138.

Step 1: sonogashira reaction of 24 with 4-methylpent-1-yn-3-ol gave tert-butyl 3-hydroxy-3- (3- (3-hydroxy-4-methylpent-1-ynyl) phenyl) propylcarbamate as a dark brown oil. Yield (1.73g, 81%):¹HNMR(400MHz,CDCl₃)7.44(s,1H),7.28-7.34(m,3H),4.86(bs,1H),4.72(bs,1H),4.39(t,J=6.0Hz,1H),3.46-3.51(m,2H),3.11-3.19(m,1H),1.78–2.04(m,4H),1.45(s,9H),1.02(d,J=7.2Hz,3H),1.06(d,J=7.2Hz,3H)。

step 2: deprotection of tert-butyl 3-hydroxy-3- (3- (3-hydroxy-4-methylpent-1-ynyl) phenyl) propylcarbamate by the method described for example 138 gave example 147 hydrochloride as a pale yellow semi-solid, except that ethyl acetate was used as the solvent. Yield (0.31g, 56%):¹HNMR(400MHz,DMSO-d₆)7.80-7.98(bs,2H),7.30-7.37(m,4H),5.44(d,J=4.8Hz,1H),4.68(bs,1H),4.13(m,1H),2.82(m,2H),1.80-1.83(m,3H),0.95(d,J=7.2Hz,3H),0.93(d,J=7.2Hz,3H)。¹³CNMR(100MHz,DMSO-d₆)146.3,130.2,129.0,128.9,126.1,122.8,91.5,84.2,69.5,66.7,36.9,36.7,34.8,18.8,18.2。ESIMSm/z248[M+1]⁺。

example 148

Preparation of 1- (2- (3- (3-aminopropyl) phenyl) ethynyl) cyclobutanol

1- (2- (3- (3-aminopropyl) phenyl) ethynyl) cyclobutanol was prepared according to the method described for example 127:

step 1: the bromide 87 was Sonogashira coupled with 1-ethynylcyclobutanol to give tert-butyl 3- (3- (2- (1-hydroxycyclobutyl) ethynyl) phenyl) propylcarbamate as a brown oil. Yield (0.32g, 61%).¹HNMR(400MHz,CDCl₃)7.11-7.30(m,4H),4.52(bs,1H),3.10-3.16(m,2H),2.62(t,J=7.6Hz,2H),2.42-2.56(m,2H),2.22-2.40(m,2H),1.78-1.91(m,4H),1.44(s,9H)。

Step 2: tert-butyl 3- (3- (2- (1-hydroxycyclobutyl) ethynyl) phenyl) propylcarbamate was deprotected with HCl/dioxane in THF to give a yellow oil after treatment. The crude product was dissolved in a small amount of methanolic ammonia (methanolic NH)₃) (2M), and used for silica columns. Purification by flash chromatography (gradient 0 to (9.5 to 0.5) methanol-ammonia) -DCM) afforded example 148 as a yellow oil. Yield (0.09g, 43%):¹HNMR(400MHz,DMSO-d₆)7.21-7.33(m,4H),2.74(t,J=7.4Hz,2H),2.64(t,J=7.6Hz,2H),2.33-2.40(m,2H),2.18-2.26(m,2H),1.72-1.86(m,4H)。¹³CNMR(100MHz,DMSO-d₆):141.5,131.1,128.9,128.7,128.5,122.6,94.6,81.6,66.6,38.6,38.3,31.5,28.9,12.8。ESIMSm/z230[M+1]⁺。

example 149

Preparation of 1- (2- (3- (3-aminopropyl) phenyl) ethynyl) cyclooctanol

1- (2- (3- (3-aminopropyl) phenyl) ethynyl) cyclooctanol is prepared according to the method of equation 23:

reaction formula 23

Step 1: to a solution of amine 86(0.68g,2.9mmol) in THF (5mL) was added ethyl trifluoroacetate (0.9mL,6mmol) at room temperature. The mixture was stirred at room temperature under inert gas overnight. After removing the solvent under reduced pressure, the reaction mixture was extracted with ethyl acetate. The organic phase was washed with water and brine, over anhydrous Na ₂SO₄Dried and concentrated under reduced pressure to give a yellow oil. Purification by flash chromatography (0-10% etoac: hexanes gradient) afforded 3 as a pale yellow oil. Yield (0.61g, 62%).¹HNMR(400MHz,CDCl₃)7.33-7.37(m,2H),7.15-7.19(m,1H),7.09-7.12(m,1H),6.24(bs,1H),3.37-3.42(m,2H),2.66(t,J=7.6Hz,2H),1.88-1.96(m,2H)。

Step 2: sonogashira coupling of bromide 3 with 1-ethynylcyclooctanol to give 2,2, 2-trifluoro-N- (3- (3- (2- (1-hydroxycyclooctyl) ethynyl) phenyl) propyl) acetamide. Yield (0.344g, 47%).¹HNMR(400MHz,CDCl₃)7.22-7.29(m,2H),7.15-7.19(m,1H),7.11-7.13(m,1H),6.23(bs,1H),3.36-3.42(m,2H),1.48-2.07(m,18H)。

And step 3: to a solution of alkyne 91(0.34g,0.8mmol) in methanol (5.0mL) was added K₂CO₃(0.25g,1.7 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure, and the residue was separated with water and ethyl acetate. The organic layer was washed with water and anhydrous Na₂SO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (0-10% (9.5-0.5 methanol-ammonia) -DCM gradient) afforded example 149 as a yellow oil. Yield (0.12g, 47%):¹HNMR(400MHz,DMSO-d₆.)7.24-7.28(m,1H),7.14-7.19(m,3H),2.56-2.62(m,4H),1.83-1.94(m,4H),1.54-1.70(m,9H),1.42-1.50(m,3H)。¹³CNMR(100MHz,DMSO-d₆):142.9,131.5,129.1,129.0,128.8,123.1,100.0,96.0,82.2,70.2,38.2,33.6,32.4,28.0,24.5,22.2。ESIMSm/z286[M+1]⁺。

example 150

Preparation of 5- (3- (3-amino-1-hydroxypropyl) phenyl) -pent-4-ynylamide

5- (3- (3-amino-1-hydroxypropyl) phenyl) -pent-4-ynylamide prepared according to the procedure described for example 138.

Step 1: the reaction of 24 with pent-4-ynoic acid amide was Sonogashira to give tert-butyl 3-hydroxy-3- (3- (5-amino-5-oxopent-1-ynyl) phenyl) propylcarbamate as a yellow oil. Yield (580mg, 78%).

Step 2: deprotection of tert-butyl 3-hydroxy-3- (3- (5-amino-5-oxopent-1-ynyl) phenyl) propylcarbamate afforded example 150 hydrochloride as a pale yellow semi-solid. Yield (110mg, 51%):¹HNMR(400MHz,DMSO-d₆)7.22-7.29(m,4H),4.63-4.66(m,1H),2.77-2.82(m,2H),2.59(t,J=7.2Hz,2H),2.33(t,J=7.2Hz,2H),1.75-1.84(m,2H),¹³CNMR(100MHz,DMSO-d₆)172.9,146.2,130.2,128.9,100.0,125.7,123.4,90.3,81.0,69.8,37.5,36.9,34.6,15.5,ESIMSm/z247[M+1]⁺。

example 151

Preparation of 3-amino-1- (3- (2-cyclooctylethynyl) phenyl) propan-1-ol

3-amino-1- (3- (2-cyclooctylethynyl) phenyl) propan-1-ol was prepared according to the method described for example 138.

Step 1: the 24 was subjected to Sonogashira reaction with ethynylcyclooctane to give 3- (3- (2-cyclooctyl) as a pale yellow oilEthynyl) phenyl) -3-hydroxypropylcarbamic acid tert-butyl ester. Yield (470mg, 91%):¹HNMR(400MHz,CDCl₃)7.39(s,1H),7.22-7.29(m,3H),4.86(bs,1H),4.72(m,1H),3.23(bs,1H),3.11-3.19(m,1H),2.76-2.79(m,2H),1.92-1.96(m,2H),1.74-1.81(m,6H),1.53-1.60(m,6H),1.45(s,9H),1.27(m,2H)。

and 5: deprotection of tert-butyl 3- (3- (2-cyclooctylethynyl) phenyl) -3-hydroxypropylcarbamate afforded example 151 hydrochloride as a light yellow semi-solid. Yield (86mg, 34%):¹HNMR(400MHz,DMSO-d₆)7.77-7.94(bs,3H),7.23-7.33(m,4H),5.60(bs,1H),4.65-4.69(m,1H),2.80-2.84(m,3H),1.77-1.89(m,8H),1.12-1.24(m,8H),¹³CNMR(100MHz,DMSO-d₆)146.1,130.2,128.9,125.6,123.6,95.6,81.0,69.6,37.0,36.7,31.5,30.4,27.4,25.3,24.4,ESIMSm/z286[M+1]⁺。

example 152

Preparation of 3-amino-1- (3- (5-methoxypent-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (5-methoxypent-1-ynyl) phenyl) propan-1-ol was prepared as described for example 138.

Step 1: sonogashira reaction of 24 with 5-methoxypent-1-yne was carried out to give tert-butyl 3- (3- (5-methoxypent-1-ynyl) phenyl) propylcarbamate as a yellow oil. Yield (280mg, 27%): ¹HNMR(400MHz,CDCl₃)7.40(s,1H),7.22-7.29(m,3H),4.86(bs,1H),4.71(m,1H),3.52(t,J=6.0Hz,2H),3.45-3.50(m,1H),3.37(s,3H),3.13-3.18(m,1H),2.49(t,J=7.2Hz,2H),1.82-1.89(m,4H),1.45(s,9H)。

Step 2: deprotection of tert-butyl 3- (3- (5-methoxypent-1-ynyl) phenyl) propylcarbamate affords a light yellow semi-solidExample 152 hydrochloride salt. Yield (151mg, 68%):¹HNMR(400MHz,DMSO-d₆)7.67(bs,2H),7.26-7.35(m,4H),5.60(bs,1H),4.65-4.69(m,1H),3.44(t,J=6.4,2H),3.25(s,3H),2.80-2.85(m,2H),2.46(t,J=7.2Hz,2H),1.72-1.87(m,4H)。¹³CNMR(100MHz,DMSO-d₆)146.1,130.2,128.9,125.7,123.4,90.4,81.2,70.9,69.7,58.4,36.9,36.7,28.7,15.9。MS:248[M+1]⁺。

example 153

Preparation of (R) -3-amino-1- (3- (4-phenylbutan-1-ynyl) phenyl) propan-1-ol

(R) -3-amino-1- (3- (4-phenylbutan-1-ynyl) phenyl) propan-1-ol was prepared according to the procedure described in equation 16 for determining the absolute stereochemistry of example 100.

Step 1: with the exception of stirring the reaction mixture at 70 ℃ for 4 hours and then at 60 ℃ for 17 hours, the aryl bromide (64) was subjected to Sonogashira coupling with 4-phenylbutyyne according to the procedure described in example 1 to give crude (R) -2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-phenylbut-1-ynyl) phenyl) propyl) acetamide as a pale yellow oil, which was used in the next step without purification. Yield (0.49g, 77%).

Step 2: deprotection of (R) -2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-phenylbut-1-ynyl) phenyl) propyl) acetamide as described in example 100 (determination of absolute stereochemistry) gave example 153 as a colorless oil. Yield (0.195g, 60%).¹HNMR(400MHz,CD₃OD)7.31-7.33(m,1H),7.15-7.29(m,8H),4.67(dd,J=8.0,5.3Hz,1H),2.87(t,J=7.2Hz,2H),2.64-2.74(m,4H),1.72-1.85(m,2H)；¹³CNMR(100MHz,CD₃OD)145.6,140.9,130.1,128.8,128.7,128.4,128.2,126.1,125.1,124.1,89.0,81.1,72.0,71.9,41.5,38.4,35.0,21.2；RP-HPLC,96.4%(AUC);LCMSm/z=280.2。

Example 154

Preparation of 4- ((3- (3-amino-1-hydroxypropyl) -5-chlorophenyl) ethynyl) hept-4-ol

4- ((3- (3-amino-1-hydroxypropyl) -5-chlorophenyl) ethynyl) heptan-4-ol was prepared according to the methods described for examples 107, 10, and 1.

Step 1: alkylation of 5-bromo-3-chlorobenzaldehyde with acetonitrile yielded 3- (5-bromo-3-chlorophenyl) -3-hydroxypropionitrile as a clear oil. Yield (3.21g, 54%):¹HNMR(400MHz,DMSO-d₆)7.62(t,J=2.0Hz,1H),7.58-7.57(m,1H),7.49-4.48(m,1H),6.18(d,J=4.8Hz,1H),4.93-4.90(m,1H),2.93(ABd,J=16.8,5.2Hz,1H),2.86(ABd,J=17.2,6.8Hz,1H)。

step 2: reacting 3- (5-bromo-3-chlorophenyl) -3-hydroxypropionitrile with BH₃THF reduction followed by amine protection to give N- (3- (5-bromo-3-chlorophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a clear oil. Yield (3.15g, 71%):¹HNMR(400MHz,DMSO-d₆)9.30(bs,1H),7.55(t,J=2.0Hz,1H),7.49-7.48(m,1H),7.394-7.387(m,1H),5.57(d,J=4.8Hz,1H),3.30-3.15(m,2H),1.86-1.70(m,2H)。

and step 3: n- (3- (5-bromo-3-chlorophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide was coupled with alkynol 20 as described in example 10 to give N- (3- (3-chloro-5- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a pale yellow oil. Yield (1.02g, 66%):

¹HNMR(400MHz,DMSO-d₆)9.30(bs,1H),7.35(t,J=1.6Hz,1H),7.30(s,1H),7.25(t,J=1.6Hz,1H),5.51(d,J=4.8,1H),5.17(s,1H),3.30-3.14(m,2H),1.85-1.69(m,2H),1.62-1.39(m,8H),0.89(t,J=7.2Hz,6H)。

and 4, step 4: n- (3- (3-chloro-5- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide is deprotected as described in example 1 to give example 154 as a pale yellow oil. Yield (0.68g, 87%):¹HNMR(400MHz,DMSO-d₆)7.33(t,J=1.6Hz,1H),7.26(s,1H),7.22(t,J=1.6Hz,1H),5.17(bs,1H),4.68-4.65(m,1H),2.63-2.53(m,2H),1.62-1.40(m,10H),0.89(t,J=7.2Hz,6H)。

example 155

Preparation of 4- ((5- (3-amino-1-hydroxypropyl) -2-fluorophenyl) ethynyl) hept-4-ol

4- ((5- (3-amino-1-hydroxypropyl) -2-fluorophenyl) ethynyl) heptan-4-ol was prepared according to the method described for example 154.

Step 1: alkylation of 3-bromo-4-fluorobenzaldehyde with acetonitrile gave 3- (3-bromo-4-fluorophenyl) -3-hydroxypropionitrile as a pale yellow oil. Yield (4.2g, 70%):¹HNMR(400MHz,DMSO-d₆)7.71(dd,J=6.8,2.0Hz,1H),7.44(ddd,J=8.4,5.2,2.4Hz,1H),7.35(t,J=8.8Hz,1H),6.08(bs,1H),4.90(s,1H),2.90(ABd,J=16.8,5.2Hz,1H),2.83(ABd,J=16.8,6.4Hz,1H)。

step 2: reacting 3- (3-bromo-4-fluorophenyl) -3-hydroxypropionitrile with BH₃THF reduction, followed by amine protection to give N- (3- (3-bromo-4-fluorophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a clear oil. Yield (4.3g, 73%):¹HNMR(400MHz,DMSO-d₆)9.31(bs,1H),7.62(dd,J=6.8,2.0Hz,1H),7.37-7.33(m,1H),7.30(t,J=8.8Hz,1H),5.48(d,J=4.4Hz,1H),4.60-4.56(m,1H),3.28-3.15(m,2H),1.84-1.71(m,2H)。

and step 3: coupling N- (3- (3-bromo-4-fluorophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide with alkynol 20To give 2,2, 2-trifluoro-N- (3- (4-fluoro-3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3-hydroxypropyl) acetamide as a pale yellow oil. Yield (1.37g, 78%):¹HNMR(400MHz,DMSO-d₆)9.31(t,J=5.0Hz,1H),7.37(dd,J=6.8,2.0Hz,1H),7.34-7.30(m,1H),7.18(t,J=9.0Hz,1H),5.41(d,J=4.8Hz,1H),5.19(s,1H),4.58-4.54(m,1H),3.28-3.16(m,2H),1.82-1.69(m,2H),1.63-1.41(m,8H),0.89(t,J=7.2Hz,6H)。

and 4, step 4: deprotection of 2,2, 2-trifluoro-N- (3- (4-fluoro-3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3-hydroxypropyl) acetamide gave example 155 as a light yellow oil. Yield (0.85g, 82%):¹HNMR(400MHz,DMSO-d₆)7.35(dd,J=6.8,2.0Hz,1H),7.31-7.27(m,1H),7.16(t,J=9.0Hz,1H),5.19(bs,1H),4.64(t,J=6.4Hz,1H),2.64-2.52(m,2H),1.63-1.42(m,10H),0.89(t,J=7.2Hz,6H)。

example 156

Preparation of 4- ((3- (3-amino-1-hydroxypropyl) -4-chlorophenyl) ethynyl) hept-4-ol

4- ((3- (3-amino-1-hydroxypropyl) -4-chlorophenyl) ethynyl) heptan-4-ol was prepared according to the procedure described for example 154.

Step 1: the 5-bromo-2-chlorobenzaldehyde is alkylated with acetonitrile to give 3- (5-bromo-2-chlorophenyl) -3-hydroxypropionitrile as a pale yellow liquid. Yield (4.42g, 75%): ¹HNMR(400MHz,DMSO-d₆)7.74(d,J=2.8Hz,1H),7.53(dd,J=8.8,2.8Hz,1H),7.39(d,J=8.8Hz,1H),6.30(d,J=4.8Hz,1H),5.13-5.09(m,1H),2.96(ABd,J=16.8,4.8Hz,1H),2.83(ABd,J=17.0,6.0Hz,1H)。

Step 2: reacting 3- (5-bromo-2-chlorophenyl) -3-hydroxypropionitrile with BH₃THF reduction followed by amine protection to give N- (3- (5-bromo-2-chlorophenyl) -3-hydroxypropyl) -2,2 as an orange oil2-trifluoroacetamide. Yield (2.6g, 43%):¹HNMR(400MHz,DMSO-d₆)9.42(bs,1H),7.67(d,J=2.4Hz,1H),7.45(dd,J=8.8,2.4Hz,1H),7.33(d,J=8.8Hz,1H),5.64(d,J=4.4Hz,1H),3.33-3.29(m,2H),1.96-1.80(m,1H),1.68-1.59(m,1H)。

and step 3: n- (3- (5-bromo-2-chlorophenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide is coupled with alkynol 20 to give N- (3- (2-chloro-5- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a pale yellow oil. Yield (1.03g, 68%):¹HNMR(400MHz,DMSO-d₆)9.42(t,J=5.6Hz,1H),7.53(d,J=2.0Hz,1H),7.34(d,J=8.0Hz,1H),7.23(dd,J=8.0Hz,2.0,1H),5.57(d,J=4.0Hz,1H),5.17(s,1H),4.87-4.82(m,1H),3.33-3.28(m,2H),1.87-1.79(m,1H),1.66-1.39(m,9H),0.89(t,J=7.2Hz,6H)。

and 4, step 4: deprotection of N- (3- (2-chloro-5- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide affords example 156 as a light yellow oil. Yield (0.77g, 98%):¹HNMR(400MHz,DMSO-d₆)7.53(d,J=2.4Hz,1H),7.34(d,J=8.0Hz,1H),7.23(dd,JJ=8.0,2.4Hz,1H),5.17(bs,1H),4.93(dd,J=8.8,2.4Hz,1H),2.72-2.63(m,2H),1.70-1.62(m,1H),1.59-1.39(m,9H),0.89(t,J=7.2Hz,6H)。

example 157

Preparation of 4- ((3- (3-amino-1-hydroxypropyl) -5-methoxyphenyl) ethynyl) hept-4-ol

4- ((3- (3-amino-1-hydroxypropyl) -5-methoxyphenyl) ethynyl) heptan-4-ol was prepared according to the procedure described for example 107.

Step 1: alkylation of 3-bromo-5-methoxybenzaldehyde with acetonitrile gave 3- (3-bromo-5-methoxyphenyl) -3-hydroxypropionitrile as a pale yellow oil. Yield (4.1g, 70)%)：¹HNMR(400MHz,DMSO-d₆)7.16-7.15(m,1H),7.04-7.03(m,1H),6.97-6.96(m,1H),6.04(d,J=4.8Hz,1H),4.87-4.83(m,1H),3.74(s,3H),2.89(ABd,J=16.4,5.2Hz,1H),2.81(ABd,J=16.8,6.8Hz,1H)。

Example 158

Preparation of 2- (3- (5-methoxypent-1-ynyl) phenoxy) ethylamine

2- (3- (5-Methoxypent-1-ynyl) phenoxy) ethylamine was prepared as described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with 5-methoxypent-1-yne to give 2,2, 2-trifluoro-N- (2- (3- (5-methoxypent-1-ynyl) phenoxy) ethyl) acetamide as a brown oil. Yield (1.2g, 57%):¹HNMR(400MHz,DMSO-d₆)7.19-7.23(m,1H),7.04(d,J=7.2Hz,1H),6.92(s,1H),6.82(dd,J=8.0,2.4Hz,1H),4.09(t,J=5.0Hz,2H),3.76-3.80(m,2H),3.52(t,J=6.2Hz,2H),3.36(s,3H),2.50(t,J=7.2Hz,2H),1.83-1.90(m,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- (5-methoxypent-1-ynyl) phenoxy) ethyl) acetamide gave example 158 as a brown oil. Yield (0.125g, 31%):¹HNMR(400MHz,DMSO-d₆)7.22-7.26(m,1H),6.91-6.96(m,3H),3.94(t,J=7.2Hz,2H),3.43(t,J=6.2Hz,2H),3.25(s,3H),2.89(t,J=5.6Hz,2H),2.45(t,J=7.2Hz,2H),1.72-1.79(m,2H)。¹³CNMR(100MHz,DMSO-d₆)158.4,129.7,124.2,123.4,116.8,114.9,89.9,80.6,70.4,69.4,57.9,40.5,28.2,15.5。ESIMSm/z234[M+1]⁺。

example 159

Preparation of 4- ((3- ((1R,2R) -3-amino-1-hydroxy-2-methylpropyl) -phenyl) ethynyl) hept-4-ol

4- ((3- ((1R,2R) -3-amino-1-hydroxy-2-methylpropyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure described for equation 24.

Reaction formula 24

Step 1: to 3-bromobenzaldehyde (22) (4.16g,22.5mmol), (R) -4-benzyl-3-propionyloxyoxazolidin-2-one (92) (5.111g,21.9mmol) and anhydrous MgCl under argon₂(0.21g,2.23mmol) in ethyl acetate (40mL) was added Et₃N (6.3mL,45.2mmol) and chlorotrimethylsilane (4.3mL,34.0 mmol). The reaction mixture was stirred at room temperature for 22 hours, then filtered through a layer of silica gel, washed with ethyl acetate. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (2-25% etoac/hexanes gradient) to afford oxazolidinone 93 as a colorless oil. Yield (9.79g, 91%). ¹HNMR(400MHz,DMSO-d₆)7.56(t,J=1.8Hz,1H),7.49(ddd,J=1.2,2.0,7.8Hz,1H),7.40(dt,J=1.2,7.6Hz,1H),7.23-7.35(m,6H),4.94(d,J=9.4Hz,1H),4.67-4.75(m,1H),4.30(t,J=8.6Hz,1H),4.12(dd,J=2.9,8.8Hz,1H),4.00-4.08(m,1H),3.02(dd,J=3.1,13.5Hz,1H),2.91(dd,J=7.4,13.5Hz,1H),0.78(d,J=7.0Hz,3H),-0.09(s,9H)。

Step 2: to LiBH₄(2M in THF, 65mL,130mmol) in ice-cold solution MeOH (2.6mL,64.2mmol) was added and the mixture was stirred at 0 deg.C for 5 min. A solution of oxazolidinone 93(9.59g,19.6mmol) in anhydrous THF (170mL) was added and the reaction mixture was stirred at 0 ℃ for 1.5 hours, then at room temperature for 1.5 hours. Slowly adding NH to the reaction mixture₄Aqueous Cl (25%,75mL) for 1 hour, followed by further addition of ethyl acetate at room temperatureAnd stirred until the mixture was clear and transparent. The layers were separated and the aqueous layer was extracted with EtOAc. The combined organic layer was washed with saturated brine, and anhydrous MgSO₄Dried, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (5-30% etoac-hexanes gradient) to afford alcohol 94 as a colorless oil. Yield (2.97g, 48%).¹HNMR(400MHz,DMSO-d₆)7.38-7.43(m,2H),7.24-7.27(m.2H),4.58(d,J=6.85Hz,1H),4.38(t,J=5.3Hz,1H),3.32-3.38(m,1H),3.22-3.29(m,1H),1.73-1.80(m,1H),0.61(d,J=6.85Hz,3H),-0.05(s,9H)。

And step 3: DEAD (1.9mL,11.4mmol) was added to alcohol 94(2.97g,9.36mmol), phthalimide (1.52g,10.3mmol) and Ph₃P (3.02g,11.5mmol) in dry THF (40mL) and the mixture stirred at room temperature for 1 h. The solvent was concentrated under reduced pressure to give an orange residue which was vigorously stirred with 10% ethyl acetate in hexane. Triphenylphosphine oxide precipitated and was removed by filtration, washing with 5% ethyl acetate in hexane. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (5-30% etoac-hexanes gradient) to give bromide 95 as a colorless oil. Yield (3.97g, 95%): ¹HNMR(400MHz,DMSO-d₆)7.76-7.80(m,4H),7.47(t,J=1.8Hz,1H),7.27-7.35(m,2H),7.22(t,J=7.8Hz,1H),4.66(d,J=5.7Hz,1H),3.63(dd,J=5.7,13.7Hz,1H),3.38(dd,J=8.8,13.5Hz,1H),2.24-2.32(m,1H),0.68(d,J=6.8Hz,3H),-0.03(s,9H)。

And 4, step 4: deprotection of 2- ((2R,3R) -3- (3-bromophenyl) -2-methyl-3- (trimethylsilyloxy) propyl) isoindoline-1, 3-dione (95) as described in example 17 gave (1R,2R) -3-amino-1- (3-bromophenyl) -2-methylpropan-1-ol (96) as a colorless oil, which was used in the next reaction without further purification.

And 5: (1R,2R) -3-amino-1- (3-bromophenyl) -2-methylpropan-1-ol (96) was protected as described in example 1. Purification by flash column chromatography (silica gel, 5% to 30% 20% etoac/hexanes gradient) afforded N- ((2R,3R) -3- (3-bromophenyl) -3-hydroxy-2-methylpropyl) -2,2, 2-trifluoroacetamide (97) as a colorless oil. The yield (1.38g,81%)：¹HNMR(400MHz,DMSO-d₆)9.21(m,1H),7.49(t,J=1.6Hz,1H),7.41(dt,J=7.2,1.6Hz,1H),7.25-7.29(m,2H),5.49(d,J=4.4Hz,1H),4.40(dd,J=6.4,4.8Hz,1H),3.23-3.29(m,1H),3.01-3.08(m,1H),1.92-2.08(m,1H),0.68(d,J=6.8Hz,3H)。

step 6: bromide 97 was coupled with 4-ethynylhept-4-ol as described in example 1 (silica gel, 40% to 60% etoac/hexanes gradient) to give 2,2, 2-trifluoro-N- ((2R,3R) -3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) -2-methylpropyl) acetamide (98) as a pale yellow oil. Yield (0.01g, 25%):¹HNMR(400MHz,MeOH-d₄)7.38(s,1H),7.27-7.30(m,3H),4.40(d,J=7.2Hz,1H),3.45(dd,J=13.6,4.2Hz,1H),3.26(dd,J=13.6,8.0Hz,1H),2.03-2.12(m,1H),1.52-1.99(m,8H),0.97(t,J=7.2Hz,6H),0.76(d,J=7.2Hz,3H)。

and 7: deprotection of 98 as described in example 1 gave example 159 as a pale yellow oil. Yield (0.04g, 45%):¹HNMR(400MHz,MeOH-d₄)7.37(s,1H),7.28-7.30(m,3H),4.39(d,J=8.0Hz,1H),2.82(dd,J=12.8,6.4Hz,1H),2.67(dd,J=12.8,6.0Hz,1H),1.79-1.88(m,1H),1.54-1.72(m,8H),0.97(t,J=7.2Hz,6H),0.76(d,J=7.2Hz,3H)。

example 160

Preparation of 1- ((3- ((1R,2R) -3-amino-1-hydroxy-2-methylpropyl) phenyl) ethynyl) cyclopentanol

1- ((3- ((1R,2R) -3-amino-1-hydroxy-2-methylpropyl) phenyl) ethynyl) cyclopentanol was prepared according to the method described for example 159.

Step 1: the bromide 97 was reacted with 1-ethynylcyclopentanol as follows, except that triethylamine was used as solvent and no phosphine (silica gel, 50-65% gradient ethyl acetate/hexane)The coupling was carried out as described in example 18 to give 2,2, 2-trifluoro-N- ((2R,3R) -3-hydroxy-3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) -2-methylpropyl) acetamide as a pale yellow oil. Yield (0.33g, 89%):¹HNMR(400MHz,MeOH-d₄)7.38(s,1H),7.25-7.33(m,3H),4.40(d,J=7.2Hz,1H),3.45(dd,J=13.2,4.2Hz,1H),3.26(dd,J=13.2,8.0Hz,1H),1.95-2.10(m,5H),1.73-1.88(m,4H),0.76(d,J=6.8Hz,3H)。

step 2: deprotection of 2,2, 2-trifluoro-N- ((2R,3R) -3-hydroxy-3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) -2-methylpropyl) acetamide was carried out as described for example 11 to give example 160 as a light yellow solid. Yield (0.07g, 41%):¹HNMR(400MHz,MeOH-d₄)7.87(s,1H),7.77-7.80(m,3H),4.99(d,J=8.0Hz,1H),3.32(dd,J=12.8,6.0Hz,1H),3.17(dd,J=12.8,6.0Hz,1H),2.22-2.54(m,9H),1.22(d,J=6.8Hz,3H)。

example 161

Preparation of (R) -3-amino-1- (3- (4-cyclohexylbut-1-ynyl) phenyl) propan-1-ol

(R) -3-amino-1- (3- (4-cyclohexylbut-1-ynyl) phenyl) propan-1-ol is prepared according to the procedure described in equation 16.

Step 1: sonogashira coupling of aryl bromide 64 with 4-cycloheylbutyyne was carried out as described in example 153 to give (R) -N- (3- (3- (4-cyclohexylbut-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a brown oil. Yield (0.672g, 88%). ¹HNMR(400MHz,DMSO-d₆)9.32(br.t,1H),7.28-7.32(m,1H),7.23-7.28(m,2H),7.18-7.23(m,1H),5.35(d,J=4.5Hz,1H),4.54(dt,J=4.9,7.4Hz,1H),3.15-3.27(m,2H),2.39(t,J=7.2Hz,2H),1.53-1.84(m,7H),1.30-1.50(m,3H),1.05-1.30(m,3H),0.78-0.92(m,2H)。

Step 2: deprotection of (R) -N- (3- (3- (4-cyclohexylbut-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide following the procedure described for example 153 gave example 161 as a colorless oil. Yield (0.377g, 75%).¹HNMR(400MHz,CD₃OD)7.33-7.35(m,1H),7.19-7.28(m,3H),4.68(dd,J=5.3,8.0Hz,1H),2.65-2.77(m,2H),2.40(t,J=7.2Hz,2H),1.62-1.90(m,7H),1.40-1.51(m,3H),1.13-1.33(m,3H),0.85-0.95(m,2H)；¹³CNMR(100MHz,CD₃OD)145.6,130.1,128.8,128.1,125.0,124.3,89.8,80.4,72.0,41.6,38.4,36.9,36.3,32.9,26.6,26.2,16.3；RP-HPLCt_R=7.65min,92.0%(AUC)；LC-MSm/z=286.4[M+H]⁺。

Example 162

Preparation of 4- ((5- (3-amino-1-hydroxypropyl) -2-methoxyphenyl) ethynyl) hept-4-ol

4- ((5- (3-amino-1-hydroxypropyl) -2-methoxyphenyl) ethynyl) heptan-4-ol was prepared according to the procedure described for example 107.

Step 1: addition of 3-bromo-4-methoxybenzaldehyde to acetonitrile gave 3- (3-bromo-4-methoxyphenyl) -3-hydroxypropionitrile as a light orange oil. Yield (10.32g, 96%):¹HNMR(400MHz,DMSO-d₆)7.58(d,J=2.0Hz,1H),7.35(dd,J=8.8,2.0Hz,1H),7.07(d,J=8.8Hz,1H),5.93(d,J=4.4Hz,1H),4.85-4.81(m,1H),3.81(s,3H),2.86(ABd,J=16.4,4.8Hz,1H),2.79(ABd,J=16.8,6.8Hz,1H)。

step 2: reacting 3- (3-bromo-4-methoxyphenyl) -3-hydroxypropionitrile with BH₃THF reduction and then amine protection to give N- (3- (3-bromo-4-methoxyphenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as an orange oil. Yield (5.76g, 40%):¹HNMR(400MHz,DMSO-d₆)9.31(bs,1H),7.49(d,J=2.0Hz,1H),7.26(dd,J=8.8,2.0Hz,1H),7.03(d,J=8.8Hz,1H),5.32(d,J=4.8Hz,1H),4.53-4.49(m,1H),3.80(s,3H),3.24-3.15(m,2H),1.79-1.72(m,2H)。

and step 3: n- (3- (3-bromo-4-methoxyphenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide was coupled with alkynol 20 as described in example 10 to give 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) -4-methoxyphenyl) propyl) acetamide as a yellow oil. Yield (0.92g, 55%): ¹HNMR(400MHz,DMSO-d₆)9.31(t,J=5.0Hz,1H),7.24-7.21(m,2H),6.95(d,J=9.2Hz,1H),5.25(d,J=4.8Hz,1H),5.05(s,1H),4.51-4.47(m,1H),3.75(s,3H),3.24-3.17(m,2H),1.77-1.72(m,2H),1.61-1.42(m,8H),0.89(t,J=7.0Hz,6H)。

And 4, step 4: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-propylhex-1-ynyl) -4-methoxyphenyl) propyl) acetamide was carried out as described for example 1 to give example 162 as a light yellow oil. Yield (0.53g, 76%):¹HNMR(400MHz,DMSO-d₆)7.22-7.19(m,2H),6.92(d,J=8.4Hz,1H),5.06(bs,1H),4.58-4.55(m,1H),3.74(s,3H),2.63-2.51(m,2H),1.64-1.42(m,10H),0.89(t,J=7.0Hz,6H)。

example 163

Preparation of 4- ((3- (3-amino-1-hydroxypropyl) -4-methylphenyl) ethynyl) hept-4-ol

4- ((3- (3-amino-1-hydroxypropyl) -4-methylphenyl) ethynyl) heptan-4-ol was prepared according to the method described for example 154.

Step 1: addition of 5-bromo-2-methylbenzaldehyde with acetonitrile gave 3- (5-bromo-2-methylphenyl) -3-hydroxypropionitrile as a pale yellow oil. Yield (3.33g, 86%):¹HNMR(400MHz,DMSO-d₆)7.61(d,J=2.0Hz,1H),7.35(dd,J=8.0,2.0Hz,1H),7.09(d,J=8.0Hz,1H),5.96(d,J=4.4Hz,1H),5.04-5.00(m,1H),2.88(ABd,J=16.8,4.4Hz,1H),2.77(ABd,J=16.8,6.4Hz,1H),2.23(s,3H)。

step 2: reacting 3- (3-bromo-2-methylphenyl) -3-hydroxypropionitrile with BH₃THF reduction and then amine protection to give N- (3- (3-bromo-2-methylphenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a light yellow oil. Yield (3.25g, 69%):¹HNMR(400MHz,DMSO-d₆)9.38(bs,1H),7.53(d,J=2.4Hz,1H),7.28(dd,J=8.0,2.4Hz,1H),7.05(d,J=8.0Hz,1H),4.73-4.70(m,1H),3.36-3.26(m,2H),2.17(s,3H),1.79-1.71(m,1H),1.68-1.59(m,1H)。

and step 3: n- (3- (3-bromo-2-methylphenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide is coupled with alkynol 20 to give 2,2, 2-trifluoro-N- (3-hydroxy-3- (5- (3-hydroxy-3-propylhex-1-ynyl) -2-methylphenyl) propyl) acetamide as a yellow oil. Yield (1.11g, 62%):¹HNMR(400MHz,DMSO-d₆)9.38(t,J=5.2Hz,1H),7.41(d,J=1.6Hz,1H),7.11(dd,J=8.0,1.6Hz,1H),7.07(d,J=8.0Hz,1H),5.26(d,J=4.4Hz,1H),5.08(s,1H),4.74-4.70(m,1H),3.35-3.25(m,2H),2.21(s,3H),1.78-1.70(m,2H),1.68-1.40(m,8H),0.89(t,J=7.2Hz,6H)。

and 4, step 4: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (5- (3-hydroxy-3-propylhex-1-ynyl) -2-methylphenyl) propyl) acetamide gave example 163 as a light yellow oil. Yield (0.71g, 85%): ¹HNMR(400MHz,DMSO-d₆)7.41(d,J=1.6Hz,1H),7.08(dd,J=7.6,1.4Hz,1H),7.05(d,J=7.6Hz,1H),5.09(bs,1H),4.83-4.80(m,1H),2.72-2.61(m,2H),2.23(s,3H),1.61-1.41(m,10H),0.89(t,J=7.0Hz,6H)。

Example 164

(E) Preparation of (E) -4- ((3- (3-aminopropyl-1-enyl) phenyl) ethynyl) hept-4-ol

(E) -4- ((3- (3-aminopropyl-1-enyl) phenyl) ethynyl) heptan-4-ol was prepared according to the methods described for examples 1 and 123.

Step 1: 4- ((3-bromophenyl) ethynyl) hept-4-ol was coupled with N-allyl-2, 2, 2-trifluoroacetamide as described for example 123 to give (E) -2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) allyl) acetamide as a light yellow oil. Yield (0.082g, 10%):¹HNMR(400MHz,DMSO-d₆)9.68(t,J=5.4Hz,1H),7.42-7.39(m,2H),7.30(t,J=8.0Hz,1H),7.23(dt,J=7.6,1.4Hz,1H),6.51(d,J=16.0Hz,1H),6.29(dt,J=16.0,6.0Hz,1H),5.13(s,1H),3.95(t,J=5.4Hz,2H),1.60-1.40(m,8H),0.89(t,J=7.2Hz,6H)。

step 2: deprotection of (E) -2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) allyl) acetamide was carried out as described for example 1 to give example 164 as a light yellow oil. Yield (0.038g, 64%):¹HNMR(400MHz,DMSO-d₆)7.36(dd,J=8.0,1.2Hz,1H),7.27(t,J=7.8Hz,1H),7.17(dt,J=7.8Hz,1.4,1H),6.45(d,J=16.0Hz,1H),6.36(dt,J=16.0,5.2Hz,1H),5.13(bs,1H),3.28(dd,J=4.0,1.0Hz,2H),1.62-1.42(m,10H),0.89(t,J=7.0Hz,6H)。

example 165

Preparation of 4- ((3- (3-aminopropyl-1-alkynyl) phenyl) ethynyl) hept-4-ol

4- ((3- (3-aminopropyl-1-ynyl) phenyl) ethynyl) heptan-4-ol was prepared according to the methods described in examples 1 and 10.

Step 1: 1, 3-dibromobenzene was coupled with alkynol 20 as described in example 10 to give 4- ((3-bromophenyl) ethynyl) hept-4-ol as a yellow liquid. Yield (4.2g, 65%):¹HNMR(400MHz,DMSO-d₆)7.56-7.52(m,2H),7.36(dt,J=8.0,1.2Hz,1H),7.30(t,J=8.0Hz,1H),5.18(s,1H),1.60-1.40(m,8H),0.89(t,J=7.2Hz,6H)。

step 2: 4- ((3-bromophenyl) ethynyl) hept-4-ol was coupled with 2,2, 2-trifluoro-N- (prop-2-ynyl) acetamide as described in example 10 to give 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) prop-2-ynyl) acetamide as a pale yellow oil. Yield (0.083g, 6%): ¹HNMR(400MHz,DMSO-d₆)10.03(t,J=5.2Hz,1H),7.40-7.33(m,4H),5.16(bs,1H),4.25(d,J=5.6Hz,2H),1.61-1.39(m,8H),0.89(t,J=7.2Hz,6H)。

And step 3: deprotection of 2,2, 2-trifluoro-N- (3- (3- (3-hydroxy-3-propylhex-1-ynyl) phenyl) prop-2-ynyl) acetamide was carried out as described in example 1 to give example 165 as a pale yellow oil. Yield (0.041g, 75%):¹HNMR(400MHz,DMSO-d₆)7.34-7.31(m,4H),5.16(s,1H),3.47(s,2H),1.77(bs,2H),1.61-1.41(m,8H),0.89(t,J=7.2Hz,6H)。

example 166

Preparation of 4- ((3- (aminomethyl) phenyl) ethynyl) hept-4-ol

4- ((3- (aminomethyl) phenyl) ethynyl) heptan-4-ol was prepared according to the methods described for examples 1 and 10.

Step 1: n- (3-bromophenyl-methyl) -2,2, 2-trifluoroacetamide is coupled with alkynol 20 as described in example 10 to give 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) benzyl) acetamide as a yellow oil. Yield (0.462g, 38%):¹HNMR(400MHz,DMSO-d₆)9.97(t,J=5.6Hz,1H),7.32(t,J=7.8Hz,1H),7.27-7.22(m,3H),5.15(bs,1H),4.35(d,J=6.0Hz,2H),1.60-1.41(m,8H),0.89(t,J=7.2Hz,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) benzyl) acetamide was carried out as described in example 1 to afford example 166 as a light yellow oil. Yield (0.254g, 78%):¹HNMR(400MHz,DMSO-d₆)7.33(s,1H),7.27-7.22(m,2H),7.18-7.16(m,1H),5.11(bs,1H),3.66(s,2H),1.97(bs,2H),1.60-1.42(m,8H),0.89(t,J=7.0Hz,6H)。

example 167

Preparation of 4- ((3- (2-aminoethyl) phenyl) ethynyl) hept-4-ol

Example 167 is an alternative synthesis of 4- ((3- (2-aminoethyl) phenyl) ethynyl) heptan-4-ol, which has been prepared in example 141. 4- ((3- (2-aminoethyl) phenyl) ethynyl) heptan-4-ol was prepared according to the methods described for examples 1 and 10.

Step 1: n- (3-bromophenylmethyl) -2,2, 2-trifluoroacetamide was coupled with alkynol 20 as described in example 10 to give 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenethyl) acetamide as a yellow oil. Yield (0.902g, 65%):¹HNMR(400MHz,DMSO-d₆)9.43(t,J=5.2Hz,1H),7.25(t,J=7.8Hz,1H),7.20-7.15(m,3H),5.11(s,1H),3.41-3.36(m,2H),2.76(t,J=7.0Hz,2H),1.61-1.40(m,8H),0.89(t,J=7.2Hz,6H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3- (3-hydroxy-3-propylhex-1-ynyl) phenethyl) acetamide was carried out as described in example 1 to give example 167 as a pale yellow oil. Yield (0.504g, 78%):¹HNMR(400MHz,DMSO-d₆)7.25-7.21(m,1H),7.17-7.14(m,3H),5.12(bs,1H),2.74-2.70(m,2H),2.58(t,J=7.2Hz,2H),1.60-1.41(m,8H),1.34(bs,2H),0.89(t,J=7.2Hz,6H)。

example 168

Preparation of 2- (3- (cyclooctylethynyl) -phenoxy) ethylamine

2- (3- (cyclooctylethynyl) -phenoxy) ethylamine was prepared according to the method described in example 18.

Step 1: the bromide 19 was Sonogashira reacted with ethynylcyclooctane to give N- (2- (3- (cyclooctylethynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide as a brown oil. Yield (0.505g, 50%):¹HNMR(400MHz,DMSO-d₆)7.21-7.27(m,1H),6.94(d,J=7.2Hz,1H),6.88-6.92(m,2H),4.09(t,J=5.4Hz,2H),3.52-3.57(m,2H),2.79-2.84(m,1H),1.86-1.92(m,2H),1.67-1.76(m,4H),1.47-1.58(m,8H)。

step 2: deprotection of N- (2- (3- (cyclooctylethynyl) phenoxy) ethyl) -2,2, 2-trifluoroacetamide afforded example 168 as a brown oil. Yield (0.071g, 48%):¹HNMR(400MHz,DMSO-d₆)7.24-7.29(m,1H),6.93-6.98(m,3H),4.13(t,J=5.0Hz,2H),3.18(t,J=5.0Hz,2H),2.77-2.81(m,1H),1.81-1.89(m,2H),1.60-1.72(m,4H),1.42-1.56(m,8H)。¹³CNMR(100MHz,DMSO-d₆)157.6,129.7,124.4,124.2,117.0,114.8,95.3,80.2,64.3,38.1,30.9,29.8,26.8,24.7,23.8。ESIMSm/z286[M+1]⁺。

example 169

Preparation of (S) -4- ((3- (2-amino-1-hydroxyethyl) phenyl) ethynyl) hept-4-ol

(S) -4- ((3- (2-amino-1-hydroxyethyl) phenyl) -ethynyl) hept-4-ol is prepared according to the procedure described in scheme 25.

Reaction formula 25

Step 1: (-) -diisopinocampheylchloroborane solution ((+) -Ipc)₂B-Cl): (+) -Ipc₂The BCl solution was prepared as described in example 100. The resulting solution was approximately 1.66M. Reduction of ketone 99 as described in example 100 gave hydroxybromide (hydroxybromide)100 as a colorless oil. Yield (0.818g, 80%):¹HNMR(400MHz,DMSO-d₆)7.57(t,J=1.8Hz,1H),7.44(ddd,J=1.0,2.0,7.8Hz,1H),7.35-7.39(m,1H),7.28(t,J=7.8Hz,1H),5.90(d,J=4.9Hz,1H),4.80(dd,J=4.7,6.5Hz,1H),3.66(ABd,J=4.5,6.5Hz,1H),3.57(ABd,J=4.5,6.5Hz,1H)。

step 2: to a solution of bromide 100(0.818g,2.92mmol) in anhydrous THF (10mL) was added a solution of potassium tert-butoxide (1M,3.5mL), the reaction mixture was stirred at room temperature for 15 minutes, concentrated under reduced pressure, and the residue was treated with water. The product was extracted twice with ethyl acetate, the organic layers were combined, brine, NH₄Washed with aqueous Cl solution and anhydrous MgSO₄Dried, filtered and the filtrate concentrated under reduced pressure to give epoxide (0.486g), which was used in the next step without purification.

Dissolve epoxide in 7N Ammonia/methanol solution (5mL) and remove NH₄Aqueous OH (25%,5mL) was added to the reaction mixture and stirred at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure to give an amine (0.801g), which was used in the next step without purification.

The amine was dissolved in anhydrous THF (5mL) and ethyl trifluoroacetate (1mL) was added. The reaction mixture was stirred at room temperature for 20 minutes, concentrated under reduced pressure, and the residue was subjected to flash chromatography to give trifluoroacetamide 101 as a colorless oil. Yield (three steps 0.608g, 67%): ¹HNMR(400MHz,DMSO-d₆)9.45(br.t,1H),7.46-7.49(m,1H),7.40-7.45(m,1H),7.25-7.30(m,2H),5.73(d,J=4.7Hz,1H),4.68(dd,J=6.7,11.3Hz,1H),3.47-3.52(m,2H)。

And step 3: coupling of 101 with 4-ethynylhept-4-ol followed by the procedure used in example 17 gave alkynol 102 as a tan oil. Yield (0.59g, 82%):¹HNMR(400MHz,DMSO-d₆)9.44(brt,J=5.5Hz,1H),7.22-7.32(m,4H),5.65(d,J=4.7Hz,1H),4.67(dd,J=11.7,6.8Hz,1H),3.25-3.31(m,2H),1.40-1.62(m,8H),0.89(t,J=7.0Hz,6H)。

and 4, step 4: a solution of alkynol 102(0.59g,1.59mmol) in ammonia/methanol (7N,10mL) and aqueous ammonia (25%,10mL) was stirred at room temperature for 70 h and concentrated under reduced pressure. Purification by flash chromatography (0-100% of 7N ammonia/methanol/dichloroethane in 10% dichloroethane) gave example 169 as a colorless oil. Yield (0.35g, 80%).¹HNMR(400MHz,CD₃OD)7.39-7.41(m,1H),7.26-7.32(m,3H),4.58(dd,J=4.7,7.6Hz,1H),2.65-2.81(m,2H),1.51-1.739m,8H),0.97(t,J=7.0Hz,6H);143.7,130.3,128.9,128.3,125.8,123.4,92.4,83.7,74.5,70.9,49.0,44.5,17.6,13.6；LC-MS:276.38[M+H]⁺；RP-HPLCtR=6.21min,98%AUC。

Example 170

Preparation of 1- (3- (3-amino-1-hydroxypropyl) phenyl) -3-methylhexan-1-yn-3-ol

1- (3- (3-amino-1-hydroxypropyl) phenyl) -3-methylhexan-1-yn-3-ol was prepared according to the method described for example 132.

Step 1: sonogashira reaction of 25 with 3-methylhexan-1-yn-3-ol gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-methylhexan-1-ynyl) phenyl) propyl) acetamide as a dark brown oil. Yield (0.611g, 55%):¹HNMR(400MHz,CDCl₃)7.41(s,1H),7.24-7.37(m,3H),4.86(m,1H),3.67-3.72(m,1H),3.38-3.44(m,1H),2.32(bs,1H),1.94-2.01(m,3H),1.71-1.76(m,2H),1.59-1.62(m,1H),1.53(s,3H),0.99(t,J=7.2,3H)。

and 5: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-hydroxy-3-methylhexan-1-ynyl) phenyl) propyl) acetamide gave example 170 as a yellow oil. Yield (0.269g, 61%):¹HNMR(400MHz,DMSO-d₆)7.61(bs,2H),7.25-7.36(m,4H),5.60(bs,1H),4.66-4.69(m,1H),2.81-2.85(m,2H),1.82-1.86(m,2H),1.59-1.63(m,2H),1.44-1.58(m,2H),1.42(s,3H),0.92(t,J=7.2,3H)。¹³CNMR(100MHz,DMSO-d₆)146.6,130.0,128.9,126.0,122.9,95.4,82.1,70.2,67.1,46.4,37.6,30.4,18.2,14.7。ESIMSm/z262[M+1]⁺。

example 171

Preparation of 3-amino-1- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) -phenyl) propan-1-ol

3-amino-1- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) -phenyl) propan-1-ol is prepared according to the procedure described for scheme 26.

Reaction formula 26

Step 1: acetonitrile addition of 3-iodobenzaldehyde (103) gave 3-hydroxy-3- (3-iodophenyl) propionitrile (104) as a yellow oil. Yield (2.58g, 55%):¹HNMR(400MHz,CDCl₃)7.82(s,1H),7.70(d,J=8.0Hz,1H),7.38(d,J=8.0Hz,1H),7.14(t,J=8.0Hz,1H),5.01(m,1H),2.80(d,J=6.4,2H),2.40(bs,1H)。

step 2: nitrile reduction of 3-hydroxy-3- (3-iodophenyl) propionitrile gave 3-amino-1- (3-iodophenyl) propan-1-ol (105) as a pale yellow oil. Yield (2.63g, quantitative yield). This compound is thus used in the next conversion process.¹HNMR(400MHz,CDCl₃)7.76(s,1H),7.58(d,J=7.6,1H),7.33(d,J=7.6,1H),7.06(t,J=8.0,1H),4.92(dd,J=8.8,2.8Hz,1H),3.09-3.14(m,1H),2.93-2.99(m,1H),1.81-1.85(m,1H),1.64-1.73(m,1H)。

And step 3: boc protection of amine 105 gave tert-butyl 3-hydroxy-3- (3-iodophenyl) propylcarbamate (106) as a yellow oil. Yield (1.39g, 40%). This compound is thus used in the next conversion process.¹HNMR(400MHz,CDCl₃)7.73(s,1H),7.58(d,J=7.6,1H),7.33(d,J=7.6,1H),7.07(t,J=8.0,1H),4.86(bs,1H),4.67(m,1H),3.45-3.51(m,2H),3.11-3.18(m,1H),1.76-1.83(m,2H),1.51(s,9H)。

And 4, step 4: sonogashira reaction of 106 with 2-ethynyltetrahydro-2H-pyran gave tert-butyl 3-hydroxy-3- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) phenyl) propylcarbamate (107) as a dark brown oil. Yield (1.21g, 83%):¹HNMR(400MHz,CDCl₃)7.45(s,1H),7.27-7.35(m,3H),4.87(bs,1H),4.69-4.71(m,1H),4.49-4.51(m,1H),4.02-4.07(m,1H),3.50-3.52(m,1H),3.56-3.61(m,1H),3.13-3.18(m,1H),1.91-1.93(m,2H),1.74-1.86(m,4H),1.51(m,2H),1.43(s,9H)。

and 5: deprotection of 107 afforded the hydrochloride salt as a yellow semi-solid which was basified and purified by flash chromatography (0-10% ((9:1) methanol-ammonia): DCM) to afford example 171 as a light yellow oil. Yield (0.273g, 85%): ¹HNMR(400MHz,DMSO-d₆)7.68(bs,2H),7.40(s,1H),7.31-7.39(m,3H),4.67-4.70(m,1H),4.51-4.54(m,1H),3.84-3.88(m,1H),3.48-3.53(m,1H),2.80-2.85(m,2H),1.79-1.88(m,4H),1.60-1.64(m,2H),1.47-1.52(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.6,130.2,129.0,128.9,126.5,122.2,89.2,85.2,69.8,66.8,65.9,37.9,37.0,32.3,25.7,21.7。ESIMSm/z260[M+1]⁺。

Example 172

Preparation of 5- (3- (3-aminopropyl) phenyl) -N-methylpent-4-yneamide

5- (3- (3-aminopropyl) phenyl) -N-methylpent-4-ynoamide was prepared according to the method described in example 127.

Step 1: sonogashira coupling of bromide 87 with N-methylpent-4-ynamide is carried out to give tert-butyl 3- (3- (5- (methylamino) -5-oxopent-1-ynyl) phenyl) propylcarbamate. Yield (1.03g, crude).¹HNMR(400MHz,CDCl₃)7.64-7.70(m,1H),7.48(d,J=6.6Hz,1H),7.21(s,1H),7.10(d,J=6.4Hz,1H),5.68(bs,1H),4.52(bs,1H),3.10-3.18(m,2H),2.85(d,J=6.8Hz,3H),2.75(t,J=7.2Hz,2H),2.60(t,J=7.6Hz,2H),2.47(t,J=7.2Hz,2H),1.75-1.82(m,2H),1.44(s,9H)。

Step 2: deprotection of tert-butyl 3- (3- (5- (methylamino) -5-oxopent-1-ynyl) phenyl) propylcarbamate using HCl in dioxane (4M) with THF as the solvent followed by neutralization with concentrated aqueous ammonia and column chromatography gave example 172 as a yellow oil. Yield (0.16g, 75%):¹HNMR(400MHz,DMSO-d₆)7.22-7.27(m,1H),7.14-7.19(m,3H),2.52-2.63(m,9H),2.34(t,J=7.2Hz,2H),1.58-1.66(m,2H)。¹³CNMR(100MHz,DMSO-d₆):171.1,143.0,131.6,129.0,128.9,128.6,123.4,90.0,81.1,41.0,34.8,34.4,32.5,25.8,15.7.ESIMSm/z245[M+1]⁺。

example 173

Preparation of 5- (3- (3-aminopropyl) phenyl) -pent-4-ynylamide

5- (3- (3-aminopropyl) phenyl) -pent-4-ynylamide prepared as described in example 127.

Step 1: sonogashira coupling of bromide 87 with pent-4-ynamide was performed to give tert-butyl 3- (3- (5-amino-5-oxopent-1-ynyl) phenyl) propylcarbamate. Yield (0.967g, crude).¹HNMR(400MHz,CDCl₃)7.64-7.70(m,1H),7.47(d,J=6.4Hz,1H),7.18(s,1H),7.10(d,J=6.4Hz,1H),5.60-5.80(m,2H),4.53(bs,1H),3.10-3.18(m,2H),2.75(d,J=7.2Hz,2H),2.60(t,J=7.6Hz,2H),2.54(t,J=7.2Hz,2H),1.76-1.80(m,2H),1.44(s,9H)。

Step 2: deprotection of tert-butyl 3- (3- (5-amino-5-oxopent-1-ynyl) phenyl) propylcarbamate using HCl in dioxane (4M) using THF as solvent followed by neutralization with concentrated aqueous ammonia and column chromatography gave example 173 as a yellow oil. Yield (0.13g, 48%): ¹HNMR(400MHz,DMSO-d₆)7.23-7.27(m,1H),7.15-7.20(m,3H),2.52-2.62(m,6H),2.33(t,J=7.4Hz,2H),1.58-1.66(m,2H)。¹³CNMR(100MHz,DMSO-d₆):172.9,143.1,131.6,129.0,128.9,128.6,123.4,90.1,81.0,41.2,34.8,34.6,32.5,15.5。ESIMSm/z231[M+1]⁺。

Example 174

Preparation of 1- (3- (3-amino-1-hydroxypropyl) phenyl) -3-ethylpent-1-yn-3-ol

1- (3- (3-amino-1-hydroxypropyl) phenyl) -3-ethylpent-1-yn-3-ol was prepared according to the method described for example 132.

Step 1: sonogashira coupling of bromide 25 with 3-ethylpent-1-yn-3-ol to give N- (3- (3- (3-ethyl-3-hydroxypent-1-ynyl) phenyl) -3-hydroxypropyl)-2,2, 2-trifluoroacetamide. Yield (0.825g, 77%).¹HNMR(400MHz,CDCl₃)7.41(s,1H),7.28-7.39(m,3H),4.83-4.87(m,1H),3.66-3.73(m,1H),3.37-3.44(m,1H),1.90-2.02(m,2H),1.70-1.81(m,4H),1.10(t,J=7.4Hz,6H)。

Step 2: deprotection of N- (3- (3- (3-ethyl-3-hydroxypent-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide affords example 174 as a yellow oil. Yield (0.52g, 91%):¹HNMR(400MHz,DMSO-d₆)7.36(s,1H),7.29-7.33(m,2H),7.26(d,J=6.4Hz,1H),4.68(t,J=4.8Hz,1H),2.74-2.82(m,2H),1.76-1.82(m,2H),1.60-1.69(m,4H),0.99(t,J=7.4Hz,6H)。¹³CNMR(100MHz,DMSO-d₆):146.5,130.1,128.9,126.0,125.0,123.0,100.0,94.0,83.3,75.0,71.0,70.2,34.5,9.2。ESIMSm/z262[M+1]⁺。

example 175

Preparation of 3-amino-1- (3- (4-cyclohexylbut-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (4-cyclohexylbut-1-ynyl) phenyl) propan-1-ol was prepared according to the method described for example 132:

step 1: the bromide 25 was Sonogashira coupled with but-3-ynylcyclohexane to give N- (3- (3- (4-cyclohexylbut-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide. Yield (1.1g, 94%).¹HNMR(400MHz,CDCl₃)7.38(s,1H),7.21-7.36(m,3H),4.84-4.88(m,1H),3.64-3.72(m,1H),3.37-3.45(m,1H),2.41(t,J=7.4Hz,2H),1.96-2.0(m,2H),1.64-1.78(m,5H),1.48-1.54(m,2H),1.36-1.46(m,1H),1.14-1.30(m,3H),0.88-1.0(m,2H)。

Step 2: implementation of deprotection of N- (3- (3- (4-cyclohexylbut-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide to give a yellow oil Example 175. Yield (0.503g, 61%):¹HNMR(400MHz,DMSO-d₆)7.22-7.29(m,3H),7.18(d,J=6.8Hz,1H),4.57(t,J=6.4Hz,1H),2.50-2.55(m,2H),2.37(t,J=7.2Hz,2H),1.54-1.70(m,7H),1.39-1.43(m,2H),1.29-1.38(m,1H),1.06-1.20(m,3H),0.80-0.90(m,2H)。¹³CNMR(100MHz,DMSO-d₆):147.3,129.7,128.9,128.6,125.7,123.4,90.8,81.1,71.3,42.6,39.2,36.8,36.2,32.8,32.8,26.6,26.2。ESIMSm/z286[M+1]⁺。

example 176

Preparation of 3-amino-1- (3- (cycloheptylethynyl) phenyl) propan-1-ol

3-amino-1- (3- (cycloheptylethynyl) phenyl) propan-1-ol was prepared according to the method described for example 19.

Step 1: the bromide 25 was Sonogashira coupled with ethynylcycloheptane (ethylcyclohexapentane) as described in example 1, followed by flash chromatography (5-40% etoac/hexanes gradient) to afford N- (3- (3- (cycloheptylethynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as an amber oil. Yield (0.507g, 51%):¹HNMR(400MHz,CDCl₃)7.42(brs,1H),7.34–7.36(m,1H),7.19–7.33(m,3H),4.81(q,J=4.0Hz,1H),3.48–3.68(m,1H),3.32–3.42(m,1H),2.74–2.82(m,1H),2.48(brs,1H),1.85–2.00(m,4H),1.70–1.80(m,4H),2.46–1.64(m,6H)。

step 2: deprotection of N- (3- (3- (cycloheptylethynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide followed by flash chromatography (5% (7N ammonia/methanol)/dichloromethane) gave example 176 as a yellow oil. Yield (0.173g, 46%):¹HNMR(400MHz,CDCl₃)7.40(s,1H),7.18–7.30(m,3H),4.91(d,J=6.8Hz,1H),3.00(brs,5H),2.74–2.82(m,1H),1.80–1.94(m,3H),1.68–1.80(m,5H),1.44–1.64(m,6H)。

example 177

Preparation of 4- ((3- (3- (methylamino) propyl) phenyl) ethynyl) hept-4-ol

4- ((3- (3- (methylamino) propyl) phenyl) ethynyl) heptan-4-ol was prepared according to the procedure described in scheme 27.

Reaction formula 27

Step 1: a mixture of allylamine carbamate 108(1.926g,12.2mmol), powdered KOH (0.734g,13.1mmol) in anhydrous DMSO (10mL) was stirred at room temperature for 5 minutes. A solution of methyl iodide (2.276g,16.03mmol) in DMSO (2mL) was then added and the reaction mixture was stirred at room temperature for 66 hours. Addition of NH ₄Aqueous Cl (25%,100mL) and the product extracted with ethyl acetate (3 × 70 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and the filtrate was concentrated under reduced pressure to give N-methylcarbamate 109 as a pale yellow liquid with a low boiling point. Yield (1.595g, 76%).¹HNMR(400MHz,CDCl₃)5.74(ddt,J=16.8,10.6,5.7Hz,1H),5.06-5.13(m,2H),3.79(d,J=5.5Hz,2H),2.80(s,3H),1.43(s,9H)。

Step 2: a solution of 1, 3-dibromobenzene (4.22g,17.9mmol) and N-methylcarbamate 109(1.555g,9.04mmol) in triethylamine (15mL) was degassed by bubbling argon for 3 minutes. Tri-o-tolylphosphine (0.140g,0.46mmol) and Pd (OAc) were then added sequentially₂(0.11g,0.49mmol), argon again bubbled for 1 minute, then vacuum/argon was applied three times. The reaction mixture was stirred at 90 ℃ under argon for 19 hours under reduced pressureConcentrating under reduced pressure. The precipitate was filtered, and the filtrate was concentrated under reduced pressure and purified by flash chromatography (5-20% etoac-hexanes gradient) to afford alkene 110 as a yellow oil. Yield (0.513g, 17%).¹HNMR(400MHz,CDCl₃)7.50(t,J=1.8Hz,1H),7.32-7.36(m,1H),7.24-7.27(m,1H),7.16(t,J=7.8Hz,1H),6.36(d,J=15.8Hz,1H),6.14(dt,J=15.8,5.9Hz,1H),3.9-4.8(m,2H),2.85(s,3H),1.46(s,9H)。

And step 3: a solution of olefin 110(0.513g,1.57mmol) in anhydrous EtOH (10mL) was degassed three times by vacuum/argon, Pd/C (10%,0.0577g) was added and stirred vigorously at room temperature for 1.5 h. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure and purified by flash chromatography (2% to 20% gradient ethyl acetate-hexanes) to give a mixture of alkanes containing debrominated alkanes as white semi-solids which was used in the next step without additional purification. Yield (0.24g, 46%).

To a mixture of alkanes (0.24g,0.73mmol) in EtOAc (5mL) was added a solution of HCl in EtOH (7.4M,1.5mL,11.1mmol) and the reaction was stirred at room temperature for 2 h. The solvent was removed under reduced pressure and the residue was dried in vacuo overnight to give amine 111 mixture containing debrominated alkane as a semi-solid, which was used in the next step without additional purification. Yield (0.19g, 93%):¹HNMR(400MHz,DMSO-d₆)8.79(brs,2H),7.43(t,J=1.6Hz,1H),7.38(dt,J=1.8,7.2Hz,1H),7.15-7.30(m,2H),2.76-2.86(m,2H),2.58-2.66(m,2H),2.46-2.51(m,3H),1.82-1.92(m,2H)。

and 4, step 4: the crude aryl bromide 111 was Sonogashira coupled with 4-ethynylhept-4-ol as described in example 17 to give example 177 as a colorless oil after two flash chromatographies (0% to 100% 10% gradient 7N ammonia/methanol-dichloromethane followed by 30% to 100% 10% gradient 7N ammonia/methanol-dichloromethane) after 21 hours of additional DMF and heating of the reaction mixture at 90 ℃. Yield (0.053g, 26%):¹HNMR(400MHz,CD₃OD)7.13-7.24(m,4H),2.61(t,J=7.6Hz,2H),2.53(t,J=7.2Hz,2H),2.34(d,J=0.8Hz,3H),1.73-1.83(m,2H),1.51-1.73(m,8H),0.97(t,J=7.0Hz,6H)；¹³CNMR(100MHz,CD₃OD)142.4,131.2,128.9,128.3,128.2,123.3,92.0,83.8,70.9,50.9,44.5,34.8,33.0,30.8,17.6,13.5；RP-HPLCt_R=6.95min,95.1%(AUC)；LC-MSm/z=288.47[M+H]⁺。

example 178

Preparation of 2- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) phenoxy) ethylamine

2- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) phenoxy) ethylamine was prepared according to the procedure described for example 18.

Step 1: the bromide 19 was Sonogashira reacted with 2-ethynyltetrahydro-2H-pyran to give 2,2, 2-trifluoro-N- (2- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) phenoxy) ethyl) acetamide as a brown oil. Yield (0.753g, 79%): ¹HNMR(400MHz,DMSO-d₆)7.26-7.32(m,1H),7.05(d,J=7.6Hz,1H),6.95-6.98(m,2H),4.50-4.53(m,1H),4.11(t,J=5.4Hz,2H),3.83-3.90(m,1H),3.54-3.58(m,2H),3.46-3.53(m,1H),1.78-1.86(m,2H).1.46-1.66(m,4H)。

Step 2: deprotection of 2,2, 2-trifluoro-N- (2- (3- ((tetrahydro-2H-pyran-2-yl) ethynyl) phenoxy) ethyl) acetamide gave example 178 as a brown oil. Yield (0.125g, 50%):¹HNMR(400MHz,DMSO-d₆)7.26-7.31(m,1H),7.02(s,1H),6.97-7.01(m,2H),4.50-4.53(m,1H),3.99(t,J=5.6Hz,2H),3.84-3.90(m,1H),3.46-3.53(m,1H),2.95(t,J=5.6Hz,2H),1.75-1.87(m,2H),1.54-1.68(m,2H),1.46-1.53(m,2H)。¹³CNMR(100MHz,DMSO-d₆)158.8,130.3,124.4,123.5,117.3,116.7,89.3,84.9,69.0,66.8,66.0,40.5,32.2,25.7,21.7。ESIMSm/z246[M+1]⁺。

example 179

Preparation of 3-amino-1- (3- (4-p-tolylbut-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (4-p-tolylbut-1-ynyl) phenyl) propan-1-ol was prepared according to the procedure described for example 132, except that the iodide of compound 25 was prepared instead of bromide.

Step 1: to a solution of 3-amino-1- (3-iodophenyl) propan-1-ol (105) (3.9g,14mmol) in DCM (50mL) was added ethyl trifluoroacetate (2mL,17mmol) and triethylamine (2.95mL,21mmol) and the mixture was stirred at room temperature for 4 h, during which time the reaction was complete. Concentrated under reduced pressure to give (2,2, 2-trifluoro-N- (3-hydroxy-3- (3-iodophenyl) propyl) acetamide) as a yellow oil. The product is sufficiently pure to be used in the next conversion step. Yield (4.9g, 93%):¹HNMR(400MHz,CDCl₃)7.71(s,1H),7.59(d,J=7.6Hz,1H),7.31(d,J=7.6Hz,1H),7.11(t,J=7.6Hz,1H),4.79-4.82(m,1H),3.67-3.72(m,1H),3.37-3.42(m,1H),2.96(bs,1H),1.87-1.99(m,2H)。

step 2: sonogashira reaction of (2,2, 2-trifluoro-N- (3-hydroxy-3- (3-iodophenyl) propyl) acetamide) with 1- (but-3-ynyl) -4-methylbenzene gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-p-tolylbut-1-ynyl) phenyl) propyl) acetamide as a yellow oil. Yield (0.310g, 60%): ¹HNMR(400MHz,CDCl₃)7.29-7.51(m,4H),7.17(d,J=8.0Hz,2H),7.13(d,J=8.0Hz,2H),4.84-4.86(m,1H),3.66-3.70(m,1H),3.38-3.43(m,1H),2.89(t,J=7.6Hz,2H),2.67(t,J=7.6Hz,2H),2.33(s,3H),2.25(bs,1H),1.92-1.97(m,2H)。

And step 3: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (4-p-tolylbut-1-ynyl) phenyl) propyl) acetamide at room temperature afforded example 179 as a white solid. Yield (0.189g, 84%):¹HNMR(400MHz,DMSO-d₆)7.24-7.31(m,3H),7.18-7.20(m,1H),7.16(d,J=8.0Hz,2H),7.10(d,J=8.0Hz,2H),4.60-4.64(m,1H),2.74-2.85(m,4H),2.63(t,J=6.8Hz,2H),2.23(s,3H),1.78-1.85(m,2H)。¹³CNMR(100MHz,DMSO-d₆)145.8,137.4,135.2,129.7,128.8,128.5,128.4,125.3,123.0,90.1,81.1,69.4,36.9,36.5,34.0,21.0,20.7。ESIMSm/z294[M+1]⁺。

example 180

Preparation of 3-amino-1- (3- (2-o-tolylbut-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (2-o-tolylbut-1-ynyl) phenyl) propan-1-ol was prepared according to the method described for example 180.

Step 1: sonogashira reaction of (2,2, 2-trifluoro-N- (3-hydroxy-3- (3-iodophenyl) propyl) acetamide) with 1- (but-3-ynyl) -2-methylbenzene gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (2-o-tolylbut-1-ynyl) phenyl) propyl) acetamide as a yellow oil. Yield (0.391g, 75%):¹HNMR(400MHz,CDCl₃)7.15-7.37(m,8H),4.84-4.86(m,1H),3.66-3.70(m,1H),3.38-3.44(m,1H),2.94(t,J=7.6Hz,2H),2.66(t,J=7.6Hz,2H),2.36(s,3H),2.32(bs,1H),1.93-1.98(m,2H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (2-p-tolylbut-1-ynyl) phenyl) propyl) acetamide at room temperature gave example 180 as a pale yellow oil. Yield (0.194g, 66%):¹HNMR(400MHz,DMSO-d₆)7.08-7.28(m,8H),4.58-4.61(t,J=6.0Hz,1H),2.83(t,J=7.2Hz,2H),2.65(t,J=7.2Hz,2H),2.55(m,2H),2.30(s,3H),1.61-1.64(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.9,138.6,135.7,130.0,129.2,129.0,128.5,128.2,126.3,125.8,125.4,122.7,89.8,81.2,70.9,42.4,31.6,19.7,19.0。ESIMSm/z294[M+1]⁺。

example 181

Preparation of 3-amino-1- (3- (3-m-tolylbut-1-ynyl) phenyl) propan-1-ol

3-amino-1- (3- (3-m-tolylbut-1-ynyl) phenyl) propan-1-ol was prepared according to the method described for example 179.

Step 1: sonogashira reaction of (2,2, 2-trifluoro-N- (3-hydroxy-3- (3-iodophenyl) propyl) acetamide) iodide with 1- (but-3-ynyl) -3-methylbenzene gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-m-tolylbut-1-ynyl) phenyl) propyl) acetamide as a brown oil. Yield (0.410g, 79%): ¹HNMR(400MHz,DMSO-d₆)9.36(bs,1H),7.22-7.33(m,3H),7.19-7.22(m,2H),7.18(s,1H),7.16(d,J=7.6Hz,1H),7.08(d,J=7.6Hz,1H),5.39(d,J=4.4Hz,1H),4.53-4.58(m,1H),3.20-3.28(m,2H),2.80(t,J=7.6Hz,2H),2.67(t,J=7.6Hz,2H),2.28(s,3H),1.75-1.83(m,2H)。

And 5: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- (3-m-tolylbut-1-ynyl) phenyl) propyl) acetamide at room temperature gave example 181 as a pale yellow oil. Yield (0.190g, 63%):¹HNMR(400MHz,DMSO-d₆)7.25-7.31(m,3H),7.19-7.22(m,2H),7.10(s,1H),7.05(d,J=7.6Hz,1H),7.0(d,J=7.6Hz,1H),4.60-4.64(m,1H),2.72-2.80(m,4H),2.65(t,J=6.8Hz,2H),2.27(s,3H),1.71-1.80(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.5,140.8,137.7,130.0,129.7,128.8,128.6,127.3,126.0,125.7,123.4,90.5,81.7,70.2,38.5,37.4,34.7,21.5,21.3。ESIMSm/z294[M+1]⁺。

example 182

Preparation of 1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclopentanol

1- ((3- (3-amino-1-hydroxypropyl) phenyl) ethynyl) cyclopentanol was prepared according to the method described for example 132.

Step 1: sonogashira reaction of bromide 25 with 1-ethynylcyclopentanol gave 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) propyl) acetamide as a brown oil. Yield (0.55g, 55%):¹HNMR(400MHz,CDCl₃)7.41(s,1H),7.28-7.35(m,3H),4.85-4.87(m,1H),3.66-3.70(m,1H),3.38-3.44(m,1H),2.41(bs,1H),1.76-2.08(m,10H)。

step 2: deprotection of 2,2, 2-trifluoro-N- (3-hydroxy-3- (3- ((1-hydroxycyclopentyl) ethynyl) phenyl) propyl) acetamide at room temperature gave example 182 as a pale yellow oil. Yield (0.126g, 31%):¹HNMR(400MHz,DMSO-d₆)7.24-7.33(m,4H),4.63-4.66(m,1H),2.71-2.83(m,2H),1.76-1.80(m,4H),1.44-1.78(m,6H)。¹³CNMR(100MHz,D₂O)143.3,131.0,128.9,128.7,126.1,122.4,117.7,92.9,82.9,74.7,71.1,41.4,36.9,35.8,22.8。ESIMSm/z260[M+1]⁺。

example 183

Preparation of 2- (4- (3- (3-amino-1-hydroxypropyl) phenyl) but-3-ynyl) phenol

2- (4- (3- (3-amino-1-hydroxypropyl) phenyl) but-3-ynyl) phenol was prepared according to the method described in example 179.

Step 1: reacting (2,2, 2-trifluoro-N- (3-hydroxy-3- (3-iodophenyl) propyl) acetamide) iodide with (2- (but-3-ynyl) phenoxy) (tert-butyl) Butyl) dimethylsilane was subjected to Sonogashira reaction to give N- (3- (3- (4- (2- (tert-butyldimethylsilyloxy) phenyl) but-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide as a brown oil. Yield (0.220g, 41%):¹HNMR(400MHz,DMSO-d₆)9.36(bs,1H),7.26-7.31(m,4H),7.19-7.21(m,1H),7.11(t,J=6.0Hz,1H),6.96(t,J=6.0Hz,1H),6.88(d,J=8.0Hz,1H),5.39(d,J=4.4Hz,1H),4.53-4.56(m,1H),3.22-3.24(m,2H),2.85(t,J=7.6Hz,2H),2.65(t,J=7.6Hz,2H),1.75-1.81(m,2H),1.0(s,9H),0.23(s,6H)。

step 2: n- (3- (3- (4- (2- (tert-butyldimethylsilyloxy) phenyl) but-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide is amine deprotected at room temperature to give example 183 as an off-white solid. Both protecting groups are removed in one step. Yield (0.087g, 72%):¹HNMR(400MHz,DMSO-d₆)7.23-7.30(m,3H),7.19(bd,J=7.2Hz,1H),6.96(bd,J=6.0Hz,1H),6.88(bt,J=8.0Hz,1H),6.71-6.76(m,2H),4.60-4.63(m,1H),2.81-2.87(m,2H),2.76(t,J=7.2Hz,2H),2.60(t,J=7.2Hz,2H),1.78-1.84(m,2H)。

example 184

Preparation of 3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol

Example 184 is an alternative synthesis of 3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol which has been prepared in example 71. 3-amino-1- (3- (4-cyclopentylbut-1-ynyl) phenyl) propan-1-ol was prepared as described for example 108.

Step 1: sonogashira reaction of 25 with but-3-ynyl-cyclopentane to give N- (3- (3- (4-cyclopentylbut-1-ynyl) phenyl) -3-hydroxypropyl as a dark brown oil) -2,2, 2-trifluoroacetamide. Yield (690mg, 69%):¹HNMR(400MHz,CDCl₃)7.22-7.57(m,4H),4.85(m,1H),3.65-3.72(m,1H),3.36-3.45(m,1H),2.41(t,J=8.0Hz,2H),2.37(bs,1H),1.90-1.97(m,3H),1.80-1.82(m,2H),1.54-1.59(m,4H),1.51-1.53(m,2H),1.12-1.15(m,2H)。

step 2: deprotection of N- (3- (3- (4-cyclopentylbut-1-ynyl) phenyl) -3-hydroxypropyl) -2,2, 2-trifluoroacetamide affords example 184 as a white semi-solid (the solvent used during salt formation was DCM rather than methanol). Yield (341mg, 68%): ¹HNMR(400MHz,DMSO-d₆)7.22-7.29(m,4H),4.63-4.66(m,1H),2.77-2.89(m,2H),2.37(t,J=7.2Hz,2H),1.76-1.89(m,3H),1.68-1.73(m,2H),1.44-1.58(m,6H),0.93-1.11(m,2H)。¹³CNMR(100MHz,DMSO-d₆)146.2,131.1,130.1,128.9,125.9,123.5,91.1,85.9,80.9,69.5,36.9,36.6,35.0,32.3,25.1,18.4。ESIMSm/z272[M+1]⁺。

Example 185

Preparation of (R) -3-amino-1- (3- (3-phenoxyprop-1-ynyl) phenyl) prop-1-ol

(R) -3-amino-1- (3- (3-phenoxyprop-1-ynyl) phenyl) propan-1-ol is prepared according to the procedure described in scheme 28.

Reaction formula 28

Step 1: to an ice-cold solution (0 ℃ C.) of vinylmagnesium bromide (1M/THF,17mL) were added anhydrous THF (10mL) and a solution of 3-iodobenzaldehyde (3.846g,16.6mmol) in anhydrous THF (12mL) in that order. The reaction mixture was stirred at 0 ℃ for 1.5 hours and NH was added₄Aqueous Cl (25%,25mL) was stirred at room temperature, the layers were separated, and the aqueous layer was extracted with ethyl acetate. NH for mixed organic layers₄Aqueous Cl, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to give allyl alcohol 112 as a pale yellow oil. Yield (4.39g, quantitative):¹HNMR(400MHz,CDCl₃)7.73(t,J=1.8Hz,1H),7.61(dt,J=1.2,7.8Hz,1H),7.30-7.34(m,1H),7.08(t,J=7.8Hz,1H),5.99(ddd,J=17.0,10.4,6.1Hz,1H),5.37(dt,J=17.0,1.2Hz,1H),5.22(dt,J=10.2,1.2Hz,1H),5.14(d,J=6.1Hz),1.9(br.s,1H)。

step 2: oxalyl chloride (1.8mL,20.6mmol) in anhydrous CH₂Cl₂(15mL) solution was cooled to-78 deg.C under argon and anhydrous DMSO (3.0mL,42.2mmol) in CH was added dropwise through an additional funnel₂Cl₂And (3) solution. Half of the DMSO solution was added first for 13 minutes and the latter half for 1 minute. The reaction mixture was stirred at-78 ℃ for 6 minutes, then allyl alcohol 112(4.39g,16.6mmol) in CH was added dropwise ₂Cl₂(15mL) of the solution was added dropwise over a period of 30 minutes. The reaction mixture was stirred at-78 ℃ for 35 minutes, then triethylamine (9mL,64.6mmol) was added dropwise over 2 minutes, and the reaction mixture was warmed to room temperature. Water (100mL) was added and, after vigorous shaking, the layers separated. By CH₂Cl₂The aqueous layer was extracted and the combined organic layers were washed successively with aqueous HCl (1%,100mL) and NaHCO₃(5%,100mL) of an aqueous solution, brine (30%,100mL), dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to give crude vinyl ketone 113(ca75% mol) as a yellow oil, which was used in the next step without purification. Yield (4.40g, quantitative):¹HNMR(400MHz,CDCl₃)8.26(t,J=1.8Hz,1H),7.87-7.92(m,2H),7.23(t,J=7.8Hz,1H),7.09(dd,J=17.0,10.7Hz,1H),6.44(dd,J=17.2,1.6Hz,1H),5.97(dd,J=10.6,1.6Hz,1H)。

and step 3: to a solution of vinyl ketone 113(3.30g,12.8mmol) and phthalimide (2.38g,16.18mmol) in anhydrous DMF (15mL) was added a solution of NaOMe (30 wt% in methanol, 0.1mL) and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure,and purified by flash chromatography (10-70% etoac-hexanes gradient) to give the crude product, which was dissolved in dichloromethane, the white precipitate was filtered off, and the filtrate was concentrated under reduced pressure. After trituration with hexane and a small amount of ethyl acetate, a white precipitate formed which was filtered and dried to give phthalimido ketone 114. Yield (2.725g, 53%): ¹HNMR(400MHz,DMSO-d₆)8.19(t,J=1.8Hz,1H),7.96(ddd,J=1.0,1.8,7.8Hz,1H),7.91(ddd,J=1.2,1.6,7.8Hz,1H),7.78-7.86(m,4H),7.30(t,J=7.8Hz,1H),3.89(t,J=7.0Hz,2H),3.39(t,J=7.0Hz,2H)。

And 4, step 4: sonogashira coupling of aryl iodide 114 with propargyl alcohol as used in example 17 gave alkynol 115 as a yellow solid. Yield (1.86g, 83%):¹HNMR(400MHz,CDCl₃)7.98(t,J=1.6Hz,1H),7.86-7.90(m,1H),7.81-7.86(m,2H),7.68-6.73(m,2H),7.59(dt,J=7.8,1.2Hz,1H),7.40(dt,J=7.8,0.4Hz,1H),4.45(s,2H),4.13(t,J=7.2Hz,2H),3.40(t,J=7.2Hz,2H),1.70(br.s,1H)。

and 5: alkynol 115 was mesylated with methanesulfonyl chloride as in example 18 to give sulfonate 116 as a brown solid. Yield (1.60g, 70%):¹HNMR(400MHz,CDCl₃)8.01(t,J=1.6Hz,1H),7.94(dt,J=1.2,8.0Hz,1H),7.81-7.87(m,2H),7.69-7.74(m,2H),7.63(dt,J=1.4,7.6Hz,1H),7.44(t,J=7.6Hz,1H),5.07(s,2H),4.13(t,J=7.2Hz,2H),3.40(t,J=7.2Hz,2H),3.15(s,3H)。

step 6: mixing phenol (0.934g,1.0mmol), mesylate 116(0.316g,0.767mmol) and K₂CO₃A mixture of (0.136g,0.984mmol) in anhydrous DMF (5mL) was stirred at room temperature under argon for 1 hour, then at 60 ℃ for 20 hours. The reaction mixture was concentrated under reduced pressure, the residue was suspended in ethyl acetate, filtered through thin layer silica gel, and washed again with ethyl acetate. The filtrate was concentrated under reduced pressure and purified by flash chromatography (5% to 40% etoac-hexanes gradient) to give phenoxypropyne 117 as a colorless oil. Yield (0.0695g, 22.1%):¹HNMR(400MHz,CDCl₃)7.97(t,J=1.6Hz,1H),7.88(dt,J=7.8,1.4Hz,1H),7.68-7.73(m,2H),7.59(dt,J=7.8,1.2Hz,1H),7.39(t,J=7.4Hz,1H),7.27-7.33(m,2H),6.95-7.04(m,3H),4.90(s,2H),4.12(t,J=7.2Hz,2H),3.38(t,J=7.2Hz,2H)。

and 7: application of Ketone 117 to (-) -Ipc₂B-Cl was reduced as described in example 100 to give (R) -hydroxyphthalimide 118 after purification by flash chromatography (5% to 30% etoac-hexanes gradient) as a colorless oil. Yield (0.0511g, 73%):¹HNMR(400MHz,CDCl₃)7.78-7.85(m,2H),7.65-7.73(m,2H),7.40-4.42(m,1H),7.19-7.34(m,5H),6.95-7.05(m,3H),4.88(s,2H),4.63(dd,J=9.0,4.5Hz,1H),3.85-3.93(m,2H),1.98-2.10(m,2H)。

and 8: phthalimide 118 was deprotected as described in example 17 except that 2 molar excess of hydrazine was used and the reaction mixture was stirred at room temperature for 70 hours to give example 185 as a colorless oil after flash chromatography (0% to 100% 10% gradient 7N ammonia/methanol-dichloroethane). Yield (0.033g, 94%): ¹HNMR(400MHz,CDCl₃)7.39-7.42(m,1H),7.24-7.36(m,5H),6.99-7.04(m,2H),6.92-6.98(m,1H),4.92(s,2H),4.70(dd,J=7.8,5.3Hz,1H),2.66-2.78(m,2H),1.74-1.89(m,2H)；RP-HPLCt_R=6.38min,97.5%(AUC);LC-MSm/z=282.52[M+H]⁺。

Example 186

Preparation of (R) -3-amino-1- (3- (3- (2, 6-dimethylphenoxy) prop-1-ynyl) phenyl) prop-1-ol

(R) -3-amino-1- (3- (3- (2, 6-dimethylphenoxy) prop-1-ynyl) phenyl) prop-1-ol was prepared according to the method described for example 185.

Step 1: alkylation of 2, 6-dimethylphenol with mesylate 116 gave 2- (3- (3- (3- (2, 6-dimethylphenoxy) prop-1-ynyl) phenyl) -3-oxopropyl) isoindoline as a colorless oil-1, 3-diketones. Yield (0.12g, 42%):¹HNMR(400MHz,CDCl₃)7.96(t,J=1.6Hz,1H),7.88(dt,J=8.0,1.2Hz,1H),7.82-7.86(m,2H),7.67-7.75(m,2H),7.58(dt,J=7.8,1.4Hz,1H),7.40(t,J=7.8Hz,1H),6.99-7.04(m,2H),6.90-6.96(m,1H),4.71(s,2H),4.13(t,J=7.4Hz,2H),3.40(t,J=7.4Hz,2H),2.35(s,6H)。

step 2: chiral reduction of 2- (3- (3- (3- (2, 6-dimethylphenoxy) prop-1-ynyl) phenyl) -3-oxopropyl) isoindoline-1, 3-dione to give (R) -2- (3- (3- (2, 6-dimethylphenoxy) prop-1-ynyl) phenyl) -3-hydroxypropyl) isoindoline-1, 3-dione as a colorless oil after purification by flash chromatography (5% to 30% etoac-hexanes gradient). Yield (0.092g, 76%):¹HNMR(400MHz,CDCl₃)7.80-7.86(m,2H),7.68-7.74(m,2H),7.39-7.41(m,1H),7.31(dt,J=7.0,1.8Hz,1H),7.20-7.28(m,2H),6.99-7.03(m,2H),6.93(dd,J=8.2,6.7Hz,1H),4.70(s,2H),4.64(dd,J=8.8,4.3Hz,1H),3.86-3.92(m,2H),2.35(s,6H),1.96-2.10(m,2H)。

and step 3: example 186 was obtained as a colorless oil after purification by flash chromatography (0% to 100% 10% gradient 7N ammonia/methanol-dichloroethane) by deprotecting (R) -2- (3- (3- (3- (2, 6-dimethylphenoxy) prop-1-ynyl) phenyl) -3-hydroxypropyl) isoindoline-1, 3-dione. Yield (0.045g, 69%):¹HNMR(400MHz,CDCl₃)7.38-7.41(m,1H),7.34(dt,J=7.4,1.6Hz,1H),7.29(t,J=7.4Hz,1H),7.26(dt,J=7.4,1.6Hz,1H),6.98-7.03(m,2H),6.91(dd,J=8.2,6.8Hz,1H),4.75(s,2H),4.70(dd,J=8.0,5.3Hz,1H),2.67-2.80(m,2H),2.32(s,6H),1.74-1.90(m,2H);RP-HPLCt_R=6.88min,99.4%(AUC);LC-MSm/z=310.68[M+H]⁺。

example 187

Preparation of (R) -3-amino-1- (3- (3- (2, 6-dimethylphenylsulfanyl) prop-1-ynyl) phenyl) prop-1-ol

(R) -3-amino-1- (3- (3- (2, 6-dimethylphenylsulfanyl) prop-1-ynyl) phenyl) prop-1-ol was prepared according to the methods described in examples 18 and 185.

Step 1: alkylation of 2, 6-dimethylthiophenol with mesylate 116 as described in example 18 gave unpurified 2- (3- (3- (2, 6-dimethylphenylsulfanyl) prop-1-ynyl) phenyl) -3-oxopropyl) isoindoline-1, 3-dione as a yellow oil, except that the reaction was carried out at room temperature for 18 hours. Yield (0.275g, 96%):¹HNMR(400MHz,CDCl₃)7.80-7.87(m,3H),7.77(t,J=1.4Hz,1H),7.68-7.73(m,2H),7.41(dt,J=1.4,7.8Hz,1H),7.33(t,J=7.8Hz,1H),7.08-7.16(m,3H),4.12(t,J=7.4Hz,2H),3.60(s,2H),3.37(t,J=7.4Hz,2H),2.58(s,6H)。

step 2: chiral reduction of 2- (3- (3- (3- (2, 6-dimethylphenylsulfanyl) prop-1-ynyl) phenyl) -3-oxopropyl) isoindoline-1, 3-dione to give (R) -2- (3- (3- (2, 6-dimethylphenylsulfanyl) prop-1-ynyl) phenyl) -3-hydroxypropyl) isoindoline-1, 3-dione as a colorless oil after purification by flash chromatography (5% to 30% etoac-hexanes gradient). Yield (0.178g, 82%):¹HNMR(400MHz,CDCl₃)7.79-7.85(m,2H),7.66-7.73(m,2H),7.23-7.27(m,1H),7.20-7.23(m,1H),7.18(t,J=7.6Hz,1H),7.07-7.14(m,4H),4.58-4.64(m,1H),3.85-3.92(m,2H),3.58(s,2H),2.58(s,6H),1.95-2.10(m,2H)。

and step 3: example 187 was obtained as a colorless oil after purification by flash chromatography (0% to 100% 10% gradient 7N ammonia/methanol-dichloroethane) by deprotecting (R) -2- (3- (3- (3- (2, 6-dimethylphenylsulfanyl) prop-1-ynyl) phenyl) -3-hydroxypropyl) isoindoline-1, 3-dione. Yield (0.090g, 71%):¹HNMR(400MHz,CDCl₃)7.25-7.29(m,1H),7.19-7.25(m,2H),7.10-7.15(m,2H),7.04-7.10(m,2H),4.66(dd,J=5.1,7.8Hz,1H),3.64(s,2H),2.64-2.77(m,2H),2.58(s,6H),1.72-1.88(m,2H);RP-HPLCt_R=7.27min,94.8%(AUC);LC-MSm/z=326.93[M+H]⁺。

example 188

Preparation of 4- ((3- (2-aminoethylamino) phenyl) ethynyl) heptan-4-ol

4- ((3- (2-aminoethylamino) phenyl) ethynyl) heptan-4-ol is prepared according to the method described in equation 29.

Reaction formula 29

Step 1: to a stirred solution of 2- (1, 3-dioxoisoindolin-2-yl) acetaldehyde (119) (3.0g,15.9mmol) in CH₂Cl₂To a solution (1000ml) was added 3-bromoaniline (120) (2.2g,13.0mmol), sodium triacetoxy borohydride (4.2g,20mmol), and acetic acid (1.2g,20 mmol). The reaction was stirred at room temperature overnight and then washed with saturated ammonium chloride, water and brine. The combined organics were dried over magnesium sulfate, filtered and concentrated in vacuo. Purification by flash chromatography (10-40% etoac/hexanes gradient) afforded benzyl bromide 121 as a yellow oil. Yield (1.7g, 38%).¹HNMR(400MHz,CDCl₃)7.80–7.88(m,2H),7.68–7.78(m,2H),6.93–6.99(m,1H),6.72–6.77(m,2H),6.49–6.54(m,1H),4.23(brs,1H),3.95(t,J=6.0Hz,2H),3.39(t,J=6.0Hz,2H)。

Step 2: benzyl bromide 121 is deprotected as described in example 17. Purification by flash chromatography (0-10% gradient (7N ammonia/methanol)/dichloromethane) afforded diamine 122 as a yellow oil. Yield (0.286g, 57%).¹HNMR(400MHz,CDCl₃)6.99(t,J=8.0Hz,1H),6.76–6.81(m,1H),6.74(t,J=2.0Hz,1H),6.49–6.54(m,1H),4.19(brs,1H),3.13(t,J=6.0Hz,2H),2.92(t,J=6.0Hz,2H),1.20(brs,2H)。

And step 3: addition of ethyl trifluoroacetate to diamine 122 as described in example 18Triflamide 123. Yield (0.462g, quantitative).¹HNMR(400MHz,CDCl₃)7.01(t,J=8.0Hz,1H),6.99(brs,1H),6.82–6.86(m,1H),6.74(t,J=2.4Hz,1H),6.50–6.55(m,1H),4.03(brs,1H),3.55(q,J=6.0,Hz,2H),3.33(t,J=6.0Hz,2H)。

And 4, step 4: the Sonogashira coupling was performed with trifluoramide 123 and 4-ethynylhept-4-ol (20) as described in example 17. Purification by flash chromatography (5-30% etoac/hexanes gradient) afforded alkynol 124 as an orange oil. Yield (0.30g, 60%). ¹HNMR(400MHz,CDCl₃)7.54(brs,1H),7.07(t,J=8.0Hz,1H),6.75–6.78(m,1H),6.61–6.64(m,1H),6.52–6.56(m,1H),3.51(q,J=5.6Hz,2H),3.32(t,J=6.0Hz,2H),1.63–1.71(m,4H),1.50–1.63(m,4H),0.95(t,J=7.2Hz,6H)。

And 5: the alkynol 124 is deprotected as described in example 1. Purification by flash chromatography (0-10% gradient (7N ammonia/methanol)/dichloromethane) afforded example 188 as a yellow solid, waxy. Yield (0.14g, 63%).¹HNMR(400MHz,DMSO)6.97–7.03(m,1H),6.46–6.54(m,3H),5.66(t,J=5.2Hz,1H),5.07(s,1H),2.94(q,J=6.4Hz,2H),2.66(t,J=6.4Hz,2H),1.38–1.62(m,10H),0.88(t,J=7,8Hz,6H)。

Example 189

Preparation of 4- ((3- (2-aminoethylthio) phenyl) ethynyl) hept-4-ol

4- ((3- (2-aminoethylthio) phenyl) ethynyl) heptan-4-ol is prepared according to the procedure described in scheme 30.

Reaction scheme 30

Step 1: alkylation of 3-bromobenzenethiol with 2-bromoethanol as described in example 18 gave 2- (3-bromobenzenethiol) ethanol (125) as an unpurified yellow oil. Yield (3.6g, 97%):¹HNMR(400MHz,CDCl₃)7.49(t,J=2.0Hz,1H),7.31(ddd,J=0.8,1.6,8.0Hz,1H),7.27(ddd,J=0.8,1.6,8.0Hz,1H),7.13(t,J=7.6Hz,1H),3.75(t,J=6.4Hz,2H),3.10(t,J=6.4Hz,2H),2.18(brs,1H)。

step 2: coupling of 2- (3-bromophenylthio) ethanol (125) with phthalimide following the procedure described in example 17 Mitsunobu followed by flash chromatography (5-20% etoac/hexanes gradient) gave 2- (2- (3-bromophenylthio) ethyl) isoindoline-1, 3-dione (126) as a white solid. Yield (4.04g, 72%):¹HNMR(400MHz,CDCl₃)7.77–7.84(m,2H),7.66–7.72(m,2H),7.50(t,J=2.0Hz,1H),7.30(ddd,J=0.8,1.6,7.6Hz,1H),7.19(ddd,J=0.8,1.6,7.6Hz,1H),7.07(t,J=7.6Hz,1H),3.93(t,J=7.2Hz,2H),3.22(J=7.2Hz,2H)。

and step 3: deprotection of 2- (2- (3-bromophenylthio) ethyl) isoindoline-1, 3-dione (126) as described in example 18 provided 2- (3-bromophenylthio) ethylamine (127) as a yellow oil. Yield (1.56g, 98%):¹HNMR(400MHz,CDCl₃)7.44(t,J=2.0Hz,1H),7.26(ddd,J=0.8,1.6,7.6Hz,1H),7.21(ddd,J=0.8,1.6,7.6Hz,1H),7.09(t,J=8.0Hz,1H),2.87–3.0(m,2H),2.82–2.86(m,2H),1.7–2.4(brs,2H)。

and 4, step 4: 2- (3-bromophenylthio) ethylamine (127) was amidated as described in example 18, followed by flash chromatography (5-20% etoac/hexanes gradient) to afford N- (2- (3-bromophenylthio) ethyl) -2,2, 2-trifluoroacetamide (128) as a colorless oil. Yield (1.75g, 80%): ¹HNMR(400MHz,CDCl₃)7.50(t,J=2.0Hz,1H),7.35(ddd,J=0.8,1.6,7.6Hz,1H),7.29(ddd,J=0.8,1.6,7.6Hz,1H),7.16(t,J=8.0Hz,1H),6.81(brs,1H),3.55(appq,J=6.4Hz,2H),3.10(t,J=6.4Hz,2H)。

And 5: alkynol 20 was reacted with N- (2- (3-bromophenylthio) ethyl) -2,2, 2-trifluoroacetamide (128) as described in example 1The method of (1) is Sonogashira coupled followed by flash chromatography (5-50% etoac/hexanes gradient) to give orange 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-propylhex-1-ynyl) thiophenyl) ethyl) acetamide (129). Yield (0.22g, 52%):¹HNMR(400MHz,DMSO)9.52–9.60(m,1H),7.26–7.36(m,3H),7.16–7.20(m,1H),5.11(s,1H),3.60(ddd,J=6.4Hz,2H),3.11(t,J=7.2Hz,2H),1.52–1.62(m,4H),1.38–1.52(m,4H),0.89(t,J=7.2Hz,6H)。

step 6: deprotection of 2,2, 2-trifluoro-N- (2- (3- (3-hydroxy-3-propylhex-1-ynyl) phenylthio) ethyl) acetamide (129) as described in example 1 followed by flash chromatography (0-10% gradient (7N ammonia/methanol)/dichloromethane) gave example 189 as a yellow oil. Yield (0.89g, 68%):¹HNMR(400MHz,MeOD)7.36–7.38(m,1H),7.32(dt,J=1.6,7.6Hz,1H),7.25(t,J=7.6Hz,1H),7.20(dt,J=1.6,7.6Hz,1H),3.01(t,J=6.4Hz,2H),2.78(brs,2H),1.62–1.74(m,4H),1.50–1.62(m,4H),0.97(t,J=7.2Hz,6H)。

example 190

Preparation of 4- ((3- (2-aminoethylsulfinyl) phenyl) ethynyl) hept-4-ol

4- ((3- (2-aminoethylsulfinyl) phenyl) ethynyl) heptan-4-ol is prepared according to the method described in reaction scheme 31.

Reaction formula 31

Step 1: to a solution of bromide 128(0.6g,1.83mmol) in acetonitrile (10mL) was added at room temperature ferric trichloride (0.015g,0.092mmol (5%)), and periodic acid (0.46g,2.0 mmol). The reaction was stirred overnight, thenWith saturated Na₂S₂O₃The reaction was quenched with aqueous solution (4 mL). The acetonitrile was removed in vacuo and the residue was extracted from water with ethyl acetate. The combined organics were washed with brine, dried over magnesium sulfate, filtered, concentrated in vacuo, and purified by flash chromatography (5-50% etoac/hexanes gradient) to afford sulfoxide 130 yield as a yellow oil (0.196g, 31%): ¹HNMR(400MHz,CDCl₃)8.37(m,1H),7.74(t,J=0.8,Hz,1H),7.61-7.65(m,1H),7.46-7.50(m,1H),7.39(t,J=8.0Hz,1H),3.78-3.89(m,1H),3.63-3.73(m,1H),3.20-3.29(m,1H),2.87–2.95(m,1H)。

Step 2: alkynol 20 was Sonogashira coupled with sulfoxide 130 as described in example 1, followed by flash chromatography (5-50% etoac/hexanes gradient) to afford trifluoroacetamide protected alkynol 131 as a yellow oil. Yield (0.15g, 67%):¹HNMR(400MHz,DMSO)8.38(brt,J=5.2Hz,1H),7.57–7.60(m,1H),7.40–7.52(m,3H),3.76–3.86(m,1H),3.62–3.72(m,1H),3.18–3.27(m,1H),2.85–2.94(m,1H),2.77(s,1H),1.62–1.74(m,4H),1.48–1.62(m,4H),0.93(t,J=7.2Hz,6H)。

and step 3: deprotection of the trifluoroacetamide protected alkynol 131 as described in example 1 followed by flash chromatography (0-10% gradient (7N ammonia/methanol)/dichloromethane) gave example 190 as a yellow oil. Yield (0.06g, 52%):¹HNMR(400MHz,MeOD)7.60–7.62(m,1H),7.43–7.51(m,2H),7.39(t,J=8.0Hz,1H),3.12–3.28(m,1H),2.98–3.12(m,1H),2.80–2.92(m,2H),1.95(brs,3H),1.60–1.72(m,4H),1.47–1.60(m,4H),0.93(t,J=7.2Hz,6H)。

example 191

Preparation of 4- ((3- (2-aminoethylsulfonyl) phenyl) ethynyl) heptan-4-ol

4- ((3- (2-aminoethylsulfonyl) phenyl) ethynyl) heptan-4-ol is prepared according to the procedure described in scheme 32.

Reaction formula 32

Step 1: to a solution of bromide 128(0.6g,1.83mmol) in ethanol (10mL) was added ammonium molybdate tetrahydrate (0.68g,0.55mmol (30%)) and hydrogen peroxide (1.9mL of a 30% solution in water, 18.3mmol) at room temperature. The reaction was stirred overnight and then saturated Na₂S₂O₃The reaction was quenched with aqueous solution (4 mL). The ethanol was removed in vacuo and the residue was extracted from water with ethyl acetate. The combined organics were washed with brine, dried over magnesium sulfate, filtered, concentrated in vacuo, and purified by flash chromatography (5-50% etoac/hexanes gradient) to give the sulfone 132 as a white solid, waxy. Yield (0.615g, 93%): ¹HNMR(400MHz,CDCl₃)8.05(t,J=2.0Hz,1H),7.81–7.87(m,2H),7.49(t,J=8.0Hz,1H),7.25(brs,1H),3.81–3.88(m,2H),3.33–3.38(m,2H)。

Step 2: the alkynol 20 was Sonogashira coupled with sulfone 132 as described in example 1, followed by flash chromatography (5-50% etoac/hexanes gradient) to afford the trifluoroacetamide protected alkynol 133 as a yellow oil. Yield (0.515g, 72%):¹HNMR(400MHz,DMSO)7.93(m,1H),7.80–7.84(m,1H),7.69–7.73(m,1H),7.55(t,J=8.0Hz,1H),7.27(brs,1H),3.79–3.86(m,2H),3.31–3.36(m,2H),1.93(brs,1H),1.64–1.78(m,4H),1.51–1.64(m,4H),0.98(t,J=7.2Hz,6H)。

and step 3: deprotection of the trifluoroacetamide protected alkynol 133 as described in example 1 followed by flash chromatography (0-10% gradient (7N ammonia/methanol)/dichloromethane) gave example 191 as a yellow oil. Yield (0.21g, 52%):¹HNMR(400MHz,MeOD)7.90–7.92(m,1H),7.78–7.82(m,1H),7.62–7.64(m,1H),7.48(t,J=8.0Hz,1H),3.20–3.25(m,2H),3.07–3.15(m,2H),1.81(brs,3H),1.62–1.75(m,4H),1.50–1.62(m,4H),0.96(t,J=7.2Hz,6H)。

example 192

Preparation of 4- ((3- (4-aminobutyl) phenyl) ethynyl) heptan-4-ol

4- ((3- (4-aminobutyl) phenyl) ethynyl) heptan-4-ol is prepared according to the procedure described in scheme 33.

Reaction formula 33

Step 1: except that the reaction mixture was heated at 90 ℃ for 18 hours, the tetrahydropyranyl bromophenol and 3-butyn-1-ol were Sonogashira coupled as described in example 17 to afford alcohol 134 after flash chromatography purification (30-100% etoac-hexanes gradient) as an orange oil. Yield (2.46g, 85%):¹HNMR(400MHz,DMSO-d₆)7.22(t,J=8.0Hz,1H),6.93–7.01(m,3H),5.45(t,J=3.2Hz,1H),4.86(t,J=5.6Hz,1H),3.66–3.75(m,1H),3.48–3.58(m,3H),2.51(t,J=6.4Hz,2H),1.64–1.90(m,3H),1.44–1.64(m,3H)。

step 2: alcohol 134 was Mitsunobu coupled with phthalimide as described in example 17, followed by flash chromatography (10-40% etoac/hexanes gradient) to afford phthalimide 135 as a colorless oil. Yield (1.77g, 84%): ¹HNMR(400MHz,CDCl₃)7.82–7.88(m,2H),7.68–7.73(m,2H),7.14(t,J=8.0Hz,1H),7.02–7.04(m,1H),6.92–6.97(m,2H),5.36(t,J=3.1Hz,1H),3.95(t,J=7.2Hz,2H),3.87(ddd,J=3.13,9.6,14.5Hz,1H),3.58(dtd,J=1.2,4.1,11.2Hz,1H),2.80(t,J=7.2Hz,2H),1.92–2.05(m,1H),1.78–1.85(m,2H),1.52–1.73(m,3H)。

And step 3: a solution of butynediophthalimide 135(1.00g,2.66mmol) in EtOH (anhydrous, 50mL) was degassed by bubbling argon for 3 minutes. Carbon supported palladium (10%,0.102g) was then added and the mixture was bubbled through argon for 30 seconds and then degassed three times using vacuum/hydrogen. The reaction mixture was stirred under a hydrogen atmosphere for 45 minutes. The mixture was filtered through filter paper to remove the catalyst and the resulting solution of 2- (4- (3- (tetrahydro-2H-pyran-2-yloxy) phenyl) butyl) isoindoline-1, 3-dione was used directly in the next step.

With the exception that the reaction mixture was heated at +50 ℃ for 16 hours, 2- (4- (3- (tetrahydro-2H-pyran-2-yloxy) phenyl) butyl) isoindoline-1, 3-dione was deprotected with hydrazine monohydrate according to the procedure described in example 17 to give 4- (3- (tetrahydro-2H-pyran-2-yloxy) phenyl) butan-1-amine as colorless oil, which was used in the next step without purification.

Deprotection of 4- (3- (tetrahydro-2H-pyran-2-yloxy) phenyl) butan-1-amine following the procedure described for example 18 gave trifluoroacetamide 136 as a colorless oil. Yield (after three steps, 0.72g, 78%):¹HNMR(400MHz,DMSO-d₆)9.37(br.t,1H),7.12–7.18(m,1H),6.75–6.83(m3H),5.40(t,J=3.2Hz,1H),3.69–3.77(m,1H),3.47–3.54(m,1H),3.17(q,J=6.4Hz,2H),2.52(t,J=7.2Hz,2H),1.63-1.90(m,3H),1.40–1.62(m,7H)。

and 4, step 4: THP-phenol 136(0.72g,2.08mmol), p-toluenesulfonic acid monohydrate (0.366g) in THF H ₂The mixture in O (3:1,20mL) was stirred at room temperature for 3.5 h. The reaction mixture was washed with NaHCO₃Brine solution treatment, separation of layers and extraction of the aqueous layer with ethyl acetate. The combined organic layers were washed with brine and concentrated under reduced pressure. Purification by flash chromatography (10-50% etoac-hexanes gradient) afforded 2,2, 2-trifluoro-N- (4- (3-hydroxyphenyl) butyl) acetamide as a colorless oil, which was used in the next step. Yield (0.438g, 96%).

To 2,2, 2-trifluoro-N- (4- (3-hydroxyphenyl) butyl) acetamide (0.438g,1.68mmol) and diisopropylethylamine (0.5mL,2.87mmol) in dry CH under argon at 0 deg.C₂Cl₂To the solution (10mL) was added a solution of trifluoromethanesulfonic anhydride (0.32mL,1.90 mmol). The reaction mixture was stirred at 0 ℃ for 15 minutes and then concentrated under reduced pressure. Ethyl acetate was added to the residue and the solution was washed with brine, dried over anhydrous magnesium sulfate, filtered and the filtrate was concentrated under reduced pressure to give crude triflate 137 as a light brown oil which was used in the next step without additional purification. Yield (0.683g, quantitative):¹HNMR(400MHz,DMSO-d₆)9.38(brt,1H),7.45(t,J=8.0Hz,1H),7.24–7.34(m,3H),3.17(q,6.4Hz,2H),2.64(t,J=7.2Hz,2H),1.40–1.60(m,4H)。

and 5: trifluoromethanesulfonate 137 was Sonogashira coupled with 4-ethynylhept-4-ol as described in example 177 to give alkynol 138 as a brown oil after flash chromatography purification (5% to 40% etoac-hexanes gradient). Yield (0.327g, 51%): ¹HNMR(400MHz,DMSO-d₆)9.38(brt,1H),7.20–7.34(m,1H),7.13–7.18(m,2H),5.10(s,1H),3.17(q,J=6.4Hz,1H),2.54(t,J=7.2Hz,2H),1.38–1.62(m,12H),0.88(t,J=7.6Hz,6H)。

Step 6: trifluoroacetamide 138 was deprotected as described in example 1 except that the reaction mixture was stirred at 50 ℃ for 3.5 hours. Purification by flash chromatography (10% to 100%7N ammonia/methanol/dichloromethane-dichloromethane) afforded example 192 as a white solid. Yield (0.175g, 71%):¹HNMR(400MHz,CD₃OD)7.12–7.24(m,4H),2.46–2.66(m,4H),1.42–1.73(m,10H),1.43–1.42(m,2H),0.97(t,J=7.2Hz,6H);¹³CNMR(100MHZ,CD₃OD),142.8,131.3,128.8,128.3,128.2,123.6,91.9,83.9,70.9,44.5,41.2,35.2,32.2,28.6,17.6,13.6;RP-HPLCt_R=7.06min,92.5%(AUC);LC-MSm/z=288.25[M+H]⁺。

example 193

In vitro isomerase inhibition assay

The ability of the alkynylphenyl-linked amine derivative compounds to inhibit visual cycle isomerase activity was determined.

The isomerase inhibition reaction is essentially carried out as described (Stecheret al, J.biol.chem.274:8577-85 (1999); see also Golczaketal, Proc.Natl.Acad.Sci.USA102:8162-67 (2005)). The source of ocular circulation isomerase is bovine retinal pigment epithelial cell (RPE) microsomal membranes.

Preparation of RPE microsomal membranes

Bovine RPE microsomal membrane extract was prepared as described (golczaketal, proc.natl.acad.sci.usa102:8162-67(2005)) and stored at-80 ℃. The crude RPE microsome extract was thawed in a 37 ℃ water bath and immediately placed on ice. 50ml of the coarse RPE bodies were placed in a 50ml Teflon-glass homogenizer (Fisher scientific, catalog No.0841416M) on ice and homogenized back and forth 10 times on ice at maximum speed powered by a DeWalt electric drill. This step was repeated until the crude PRE microsome solution was homogenized. The homogenates were then spun down (50.2Ti rotor (Beckman, Fullerton, Calif.), 13,000RPM;15360Rcf) for 15 minutes at 4 ℃. The supernatant was collected and pelleted by centrifugation (160,000Rcf;50.2Ti rotor) at 42,000RPM for 1 hour at 4 ℃. The supernatant was removed and the small particles were suspended in 10mM cold MOPS buffer (final volume 12ml), pH 7.0. The resuspended RPE film in a 5 ml aliquot was homogenized to high uniformity in a glass-glass homogenizer (fisher scientific, catalog No. k885500-0021). Protein concentrations were quantified using the BCA protein assay according to the manufacturer's instructions (Pierce, Rockford, IL). The prepared homogeneous RPE was stored at-80 ℃.

Isolation of human Apo intracellular retinaldehyde-binding protein (CRALBP)

The recombinant human Apo intracellular retinaldehyde-binding protein (CRALBP) was cloned and expressed according to standard methods in molecular biology techniques (see Crabb et al, protein science7:746-57(1998); Crabb et al, J.biol.chem.263:18688-92 (1988)). Briefly, total RNA was prepared from fused ARPE19 cells (American TypeCultureCo., Manassas, Va.) using oligo (dT)_12-18Precursor Synthesis cDNA, which encodes CRALBP, is then amplified by two sequential polymerase chain reactions (see Crabb et al, J.biol. chem.263:18688-92(1988); Intres et al, J.biol. chem.269:25411-18(1994); GenBank accession No. L34219.1). The PCR product was subcloned into pTrcHis2-TOPOTA vector according to the manufacturer's instructions (Invitrogen Inc., Carlsbad, CA; catalogno. K4400-01) and then sequenced according to standard nucleotide sequence techniques. Recombinant 6 × histidine-tagged human CRALBP was expressed as OneShotTOP10 chemical E.coli cells (Invitrogen) and the recombinant polypeptide was isolated from E.coli cell lysates by nickel affinity chromatography using Ni Sepharose XK16-20 column for HPLC (Amersham bioscience, Pittsburgh, Pa.; catalogno. 17-5268-02). Purified 6 × histidine-tagged human CRALBP was dialyzed against 10 mMbis-tris-propane (BTP) and analyzed by SDS-PAGE. The molecular weight of the recombinant human CRALBP is about 39 kDal.

Isomerase assay

The amine derivative compound having an alkynylphenyl group attached and the control compound were reconstituted to 0.1M by addition to ethanol. It was prepared as a 10-fold serial dilution of each compound in ethanol (10)^-2、10^-3、10^-4、10^-5、10^-6M) for the detection of an isomerase assay.

The isomerase assay was performed in 10mM bis-tris-propane (BTP) buffer, pH7.5, 0.5% BSA (diluted in BTP buffer), 1mM sodium pyrophosphate, 20. mu.M all-trans retinol (in ethanol), and 6. mu.Mapo-CRALBP. Test compound (2 μ l) (final 1/15 dilution of serial dilution stock) was added to the above reaction mixture to which RPE microsomes were added. The same volume of ethanol was added to the control reaction (no test compound). Bovine RPE microsomes (9 μ Ι) were then added (see above) and the mixture was transferred to the initial reaction (total volume =150 μ Ι) at 37 ℃. The reaction was terminated after 30 minutes by adding methanol (300. mu.l). Heptane (300 μ l) was added and mixed with the reaction mixture by pipette. The retinoid was extracted by stirring the reaction mixture followed by centrifugation in a microcentrifuge. The upper organic phase was shifted to an HPLC vial and then purified by HPLC using an Agilent1100HPLC system with a conventional phase chromatography column: SILICA (Agilent technologies, dp 5. mu.4.6 mmX,25CM; procedure flow rate 1.5 ml/min; injection volume 100. mu.l) analysis. The solvent components were 20% 2% isopropanol in ethyl acetate and 80% 100% hexane.

A₃₁₈The area under the nm curve represents the 11-cis retinol peak, which was calculated by the Agilent chemical workstation software and recorded manually. Using GraphPadprism4 software (Irvine, CA) computing IC₅₀Values (concentration of compound that inhibits 50% of 11-cis retinol formation in vitro). All experiments were performed in duplicate.

The concentration-dependent effect of the compounds disclosed herein on retinol isomerization reactions can also be evaluated using a recombinant human enzyme system. In particular, the in vitro isomerase assay was performed substantially as described (Golczak et al 2005, PNAS102: 8162-. Homogenates of HEK293 cell clones expressing recombinant human RPE65 and LRAT were the source of visual enzymes and exogenous all-trans-retinol (approximately 20 μ M) was used as substrate. Recombinant human CRALBP (about 80. mu.g/mL) was added to enhance the formation of 11-cis retinal. The 200. mu. LBis-Tris phosphate buffer (10mM, pH7.2) based on the reaction mixture also contained 0.5% BSA and 1mM Na PPi. In this analysis, the reaction was repeated at 37 ℃ for 1 hour and stopped by adding 300. mu.L of methanol. The amount of the reaction product 11-cis-retinol was measured by HPLC analysis followed by heptane extraction of the reaction mixture. Recording the Peak Area Unit (PAU) associated with 11-cis-retinol in HPLC chromatogram and recording IC by GraphPadprism ₅₀Concentration dependence of the values. The ability of the compounds disclosed herein to inhibit isomerization reactions was quantified and their IC's determined separately₅₀The value is obtained. The following table summarizes the IC's of various compounds of the present invention₅₀A value determined by either of the two methods described above. FIG. 1 is a representative graph of the determination of the compound IC50 of example 2. Additional human and bovine IC50s in vitro data are shown in tables 14A and 14B.

TABLE 14A in vitro inhibition data

TABLE 14 bovine in vitro inhibition data

Example 194

In vivo murine isomerase assay

The ability of the alkynylphenyl-linked amine derivatives to inhibit isomerase was determined by in vivo murine isomerase analysis. Brief exposure of the eye to intense light ("bleaching" or simply "bleaching" of the visual pigment) is known to photo-isomerize almost all of the 11-cis-retinal in the retina. The recovery of bleached 11-cis-retinal can be used to assess isomerase activity in vivo. Delayed recovery, represented by low 11-cis-retinal oxime levels, indicates inhibition of the isomerization reaction. The procedure was generally performed as described by Golczak et al, Proc.Natl.Acad.Sci.USA102:8162-67 (2005). Also see Deigner et al, Science,244:968-71 (1989); gollapalli et al, BiochimBiophysa acta.1651:93-101(2003); Parish et al, Proc.Natl.Acad.Sci.USA,14609-13(1998); Radu et al, ProcNatl Acad Sci USA101:5928-33 (2004).

6-week old, dark-adapted CD-1 (albino) male mice were gavaged orally with a compound (0.03-3 mg/kg) dissolved in 100. mu.l corn oil containing 10% ethanol (5 animals per group). Mice were gavaged with the alkynylphenyl derivative compounds described in examples 2, 18, 19, 100 and 101. After 2-24 hours in the dark, mice were exposed to 5,000 lux of white light for 10 minutes for photobleaching. Mice were allowed to recover for 2 hours in the dark. The animals were then sacrificed by carbon dioxide inhalation. Retinoids were extracted from the eye and the regeneration of 11-cis-retinal was assessed at various time intervals.

Extraction of ocular retinoids

All steps are performed with minimal red illumination in the dark (requiring low light darkroom illumination and a filtered red flash lamp for local illumination) (see, e.g., Maeda et al, J.neuro. chem85: 944-. Immediately after sacrifice, the eyeballs were removed and placed in liquid nitrogen for storage.

The eyeballs were placed in 500. mu.L of bis-tris-propane buffer (10mM, pH 7.3) and 20. mu.L of 0.8 Mhydroxilamine (pH 7.3). The eye was cut into small pieces with the iris and vigorously homogenized in the tube with a mechanical homogenizer (polytron pt1300D) at 30000rpm until no visible tissue remains. To each tube was added 500. mu.L of methanol and 500. mu.L of heptane. The tube was placed in a vortex apparatus (vortex) to allow the contents to mix thoroughly for 15 minutes at room temperature. The organic phase was separated from the aqueous phase by centrifugation at 13Krpm for 10 minutes at 4 ℃. Remove and transfer 240 μ L of solution from the upper layer (organic phase) with a glass pipette into a clean 300 μ L glass liner inserted inside the HPLC vial and close-off the vial.

On an Agilent1100HPLC system with a conventional phase chromatography column: SILICA (BeckmanCoultier, dp5 μm,4.6mMx250mM) analyzed samples. The run method had a flow rate of 1.5 ml/min; the solvent components were 15% solvent 1 (1% isopropanol in ethyl acetate), and 85% solvent 2(100% hexane). The injection volume for each sample was 100 μ l; the detection wavelength was 360 nm. The area under the curve for 11-cis retinal oxime was calculated and manually recorded by the Agilent chemical workstation software. Data processing was performed using Prizm software.

Positive control mice (no compound administered) were sacrificed under full dark adaptation and the eyeballs were analyzed for retinoids. The photo (bleached) control mice (no compound applied) were sacrificed and the retinoids were isolated and analyzed immediately after photo-treatment.

In vivo Using the Compounds of examples 2, 18, 19, 100 and 101 (Compounds 2, 18, 19, 100 and 101)Study of isomerase inhibition dose response. Male Balb/c mice (8/group) were orally administered compound 2-HCl, compound 18-HCl, compound 100-HCl or compound 101-HCl in sterile aqueous solutions at doses of 0.03, 0.1, 0.3, 1 and 3mg/kg and photobleached for 4 hours after dosing. Animals receiving compound-19 HCl were administered orally a sterile aqueous solution of the compound at doses of 0.01 and 1mg/kg and photobleached 4 hours after dosing. Recovery and retinoid analysis were performed as described above. Dark control mice were vehicle-only treated, sacrificed without light treatment under full dark adaptation, and analyzed. The concentration-dependent inhibition of isomerase activity 4 hours after administration of compounds 2, 18 and 19 is shown in FIGS. 2-4. The inhibition of 11-cis retinal (oxime) recovery by compounds 2, 18, 19, 100 and 101 are shown in FIGS. 5-9, respectively. Estimated ED ₅₀(the dose of compound capable of inhibiting 50% of the recovery of 11-cis retinal (oxime)) is shown in Table 15B.

Time course studies were performed to determine the isomerase inhibitory activity of compounds 2, 18 and 19. Male Balb/c mice (4/group) received 3mg of Compound 2-HCl, Compound 18-HCl or Compound 19-HCl (in water) per kg body weight orally as gavage. Animals were then "photo-bleached" (5000 lux of white light for 10 minutes) at 2, 4, 8, 16 and 24 hours post-application and returned to the dark to restore 11-cis-retinal content in the eye. Mice were sacrificed 2 hours after bleaching, eyes were removed, and retinoid content was analyzed by HPLC. The results are shown in FIGS. 10 to 12.

A single dose study of the compound of example 36 (compound 36) was performed at 1mg/kg and 5mg/kg oral dosing 2, 4, 6 and 24 hours after bleaching. Experiments were performed in CD1 male mice. The results were analyzed by HPLC. The results are shown in FIGS. 13(1mg/kg) and 14 (data at 5 mg/kg), and in Table 15C. Example 36 was inactive at 1mg/kg (FIG. 13) and at 5mg/kg was-50% active at 2 and 6 hours, with complete recovery at 24h (FIG. 14).

TABLE 15A

In vivo inhibition data

TABLE 15B

In vivo inhibition data

Example numbering	ED50
		2	0.69
18	1.73
		19	0.33
100	0.2
		101	0.38

TABLE 15C

In vivo inhibition data

Example 195

Preparation of retinal nerve cell culture system

This example describes a method for long-term culture of retinol neural cells.

All compounds and reagents were purchased from SigmaAldrich chemical corporation (St. Louis, Mo.), except where specifically noted.

Retinal nerve cell culture

Porcine eyeballs were obtained from KapowsinMeats, inc. (Graham, WA). The eye contents are removed and the muscles and tissues are cleared from the eye. The neuroretina is dissected in half along the equator of the eyeball and dissected from the anterior half of the eyeball in buffered saline solution according to standard methods known in the art. Briefly, a piece of retina, ciliary body, and vitreous is cut from the anterior half of the eyeball, and the retina is gently cut from the clear vitreous. Each retina was isolated with papain (worthington biochemical corporation, Lakewood, NJ), then inactivated with Fetal Bovine Serum (FBS) and DNaseI added at 134Kunitz units/ml. The enzymatically dissociated cells were ground and collected by centrifugation and suspended in Dulbecco's Modified Eagle Medium (DMEM)/F12 medium (GibcoBRL, Invitrogen Life technologies, Carlsbad, Calif.) containing 25. mu.g/ml insulin, 100. mu.g/ml transferrin, 60. mu.M putrescine, 30nM selenium, 20nM progesterone, 100U/ml penicillin, 100. mu.g/ml streptomycin, 0.05MHepes and 10% FBS. The isolated primary retinal cells were placed on poly-D-lysine-and matrigel (BD, Franklin lakes, NJ) coated glass coverslips and placed on 24-well tissue culture plates (falcon tissue cultures, Fisher scientific, Pittsburgh, Pa.). At 37 ℃ and 5% CO ₂Next, the cells were maintained in 0.5ml of medium (as described above, except that it contained only 1% FBS) for 5 days to 1 month.

ImmunocytochemistryChemical analysis

Retinal nerve cells were cultured for 1, 3, 6, and 8 weeks and cells were analyzed by immunocytochemistry at each time point. Immunocytochemical analysis was performed according to standard techniques well known in the art. Rod cell photoreceptors were identified by labeling with rhodopsin-specific antibodies (monoclonal mouse, diluted 1: 500; Chemicon, Temecula, CA). Medium heavy neurofilament antibodies (NFM rabbit polyclonal, diluted 1:10,000, Chemicon) were used to identify ganglion cells; beta 3-tubulin antibody (G7121 mouse monoclonal, diluted to 1:1000, Promega, Madison, Wis.) was used universally for determination of interneurons and ganglion cells, and calbindin antibody (AB1778 rabbit polyclonal, diluted to 1:250, Chemicon) and calretinin antibody (AB5054 rabbit polyclonal, diluted to 1:5000, Chemicon) were used to determine the subspecies of calbindin-and calretinin-expressing interneurons. Briefly, retinal cell cultures were mixed with 4% paraformaldehyde (Polysciences, Inc, Warrington, PA) and/or ethanol, washed in Dulbecco Phosphate Buffered Saline (DPBS), and incubated with primary antibody for 1 hour at 37 ℃. Cells were then washed with DPBS, incubated with secondary antibody (secondary antibody binding to Alexa 488-OR Alexa568- (molecular probes, Eugene, OR)) and washed with DPBS. Nuclei were transfected with 4', 6-diamidino-2-phenylindole (DAPI, molecular probe), cultures were washed with DPBS before removing the glass coverslips and mounted with fluorocount-G (southern biotech, Birmingham, AL) slides for observation and analysis.

Survival of mature retinal neurons in culture after various times was determined by histochemical analysis. Determining photoreceptor cells using rhodopsin antibodies; determining ganglion cells using the NFM antibody; and determining amacrine and horizontal cells by transfection with antibodies specific for calretin.

Cultures were analyzed by counting rhodopsin-labeled photoreceptors and NFM-labeled ganglion cells using an OlympusIX81 or CZX41 microscope (Olympus, tokyo, japan). The 20-fold field of each coverslip was counted with a 20 × objective. The 6 coverslips for each condition in each experiment were analyzed by this method. Cells not exposed to any stressor are counted and cells exposed to stressor are normalized to the number of control cells.

Example 196

Effect of alkynyl phenyl derivative Compounds on retinal cell survival

This example describes the use of a mature retinal cell culture system that includes a cell stressor for testing the effect of alkynyl phenyl derivative compounds on retinal cell viability.

Retinal cell cultures were prepared as described in example 195. A2E was added as a retinal cell stressor. After 1 week of cell culture, chemical stress, A2E, was applied. A2E was diluted in ethanol and added to the retinal cell cultures at concentrations of 0, 10. mu.M, 20. mu.M and 40. mu.M. Cultures were treated for 24 and 48 hours. A2E was obtained from Dr.KojiNakanishi (Columbia university, New YorkCity, NY) or synthesized according to the method of Parish et al (Proc.Natl.Acad.Sci.USA95:14602-13 (1998)). Next, an alkynylphenyl derivative compound is added to the culture. Alkynyl phenyl derivative compounds were added to other retinal cell cultures prior to stressor application or simultaneously with the addition of A2E to the retinal cell cultures. Cultures were stored at 37 ℃ and 5% CO during stress ₂In the tissue culture incubator below. The cells were then analyzed by immunocytochemistry as described in example 133.

Apoptosis assay

Retinal cell cultures were prepared and cultured for 2 weeks as described in example 195, then exposed to 6000 lux of white light stress for 24 hours, followed by a 13 hour resting period. The device is constructed to deliver light of a particular wavelength uniformly into a particular well of a 24-well plate. The device includes a fluorescent cold white light bulb (GEP/NFC12T9/CW) wired to an AC power source. The bulb is mounted in a standard tissue culture incubator. White light stimulation was applied by placing the cell plate directly under a fluorescent bulb. CO 2₂Level ofThe temperature at the cell plate was maintained at 37 ℃ and maintained at 5%. A thin thermocouple was used to monitor the temperature. The light intensity of all devices was measured and adjusted using a photometer (P/N401025; Waltham, MA) available from Extech instruments. The alkynyl phenyl linked amine derivative compound was added to the wells of the plate prior to exposure of the cells to white light and to the other wells of the culture after exposure to white light. To assess apoptosis, TUNEL was performed as described herein.

Apoptosis assays were also performed after exposing retinal cells to blue light. Retinal cell cultures were cultured as described in example 195. After 1 week of cell culture, blue light stimulation was applied. Blue light was delivered by a custom made light source consisting of two arrays of 24 (4 x 6) blue light emitting diodes (sunbrittledp/NSSP-01 TWB7UWB12) designed for each LED per well of a disposable 24 well plate filled with cells. The first row was placed on top of a 24-well plate filled with cells, while the second row was placed on the bottom of the cell plate, so that both rows simultaneously provided light stimulation to the cell plate. The entire instrument was placed in a standard tissue culture incubator. CO 2 ₂The level was maintained at 5% and the temperature at the cell plate was maintained at 37 ℃. The temperature was monitored with a thin thermocouple. The current to each LED is controlled separately by a separate voltmeter so that the light output of all LEDs is uniform. The cell plates were exposed to 2000 luxes of blue light stimulation for 2 hours or 48 hours, followed by a 14 hour rest period. The amine derivative compound with the alkynylphenyl group attached was added to the wells of the plate prior to exposure of the cells to blue light and to the other wells of the culture after exposure to blue light. To assess apoptosis, TUNEL was performed as described herein.

To assess apoptosis TUNEL was performed according to standard techniques in the art and according to the manufacturer's instructions. Briefly, retinal cell cultures were first mixed with 4% paraformaldehyde and then ethanol, followed by rinsing in DPBS. The immobilized cells were incubated with TdT enzyme (0.2 units/. mu.l final concentration) in a reaction buffer (Fermentas, Hanover, Md.) coupled to Chroma-TideAlexa568-5-dUTP (0.1. mu.M final concentration) (molecular probes) for 1 hour at 37 ℃. The cultures were washed with DPBS and incubated with primary antibody either overnight at 4 ℃ or 1 hour at 37 ℃. Cells were then washed with DPBS, incubated with Alexa 488-linked secondary antibody, and washed with DPBS. Nuclei were transfected with DAPI, cultures were washed with DPBS before removing the glass coverslips and mounted with fluorocount-G slides for observation and analysis.

Cultures were analyzed by counting TUNEL-labeled nuclei using an Olympusix81 or CZX41 microscope (Olympus, Tokyo, Japan). The 20-fold field of each coverslip was counted with a 20 × objective. 6 coverslips per condition were analyzed by this method. Cells not exposed to the alkynylphenyl derivative compound were counted and cells exposed to the antibody were normalized to the number of cells of the control. Data were analyzed using unpaired student's t-test.

Example 197

Mouse model of light injury in vivo

This example describes the effect of an alkynyl phenyl linked amine derivative in an in vivo photodamaged mouse model.

Eye exposure to intense white light can cause photodamage to the retina. The extent of damage following light treatment can be assessed by measuring the content of cytoplasmic histone-associated DNA fragments (mono-and oligonucleosomes) in the eye (see, e.g., Wenzelet, prog.Retin. EyeRes.24:275-306 (2005)).

Dark-adapted male Balb/c (albino, 10/group) mice were gavaged with different doses (0.03, 0.1, 0.3, 1 and 3mg/kg) of the compound or vehicle only. Six hours after dosing, animals were light treated (8,000 lux white light for 1 hour). Mice were sacrificed 40 hours after recovery in the dark and retinas were excised. Cell death assay ELISA analysis was performed according to the protocol (Roche applied science, CellDeathDetectionELISAplusKit). The amount of fragment DNA within the retina was measured to estimate the retinoid-protecting activity of the compounds.

Example 198

Electroretinogram (ERG) study

This example describes the determination of the effect of alkyne-derived compounds as modulators of visual circulation on a measure of ERG response in the mouse eyeball following oral administration of the compounds to animals. The level of ERG response in the eye is determined 18 and 66 hours after administration of the compound to the animal.

Three groups of nine-week-old mice (19-25 g) of both sexes were reared at room temperature, 72 ± 4 ° f and about 25% relative humidity (C57BL/6 line, charles river laboratory, Wilmington, MA). Animals were kept in a 12 hour light/dark cycle environment, with free access to food and water and were checked for general condition and health levels prior to use in the study. Body weights were determined as representative samples prior to initial dosing. The average body weight determined from this sample was used to determine the dose for all mice in the study.

Each test compound was dissolved in a control solvent (EtOH) and diluted 1:10(90mL/900mL) in corn oil (crisco pure corn oil, j.m. smucker company, Orrville, OH) to dissolve the desired dose (mg/kg) in the desired volume (-0.1 mL/animal). The control vehicle was ethanol corn oil (1:10(0.9ml/9 ml)). Treatment protocols and animal assignments are described in table 16.

TABLE 16

^*The total results of 2 studies (n =4, n =3, respectively)

Animals were dosed orally once during the course of the light cycle (between 30 minutes and 3 hours and 30 minutes after the start of the light cycle) by gavage with the indicated vehicle control or test compound. The volume of the administered dose does not exceed 10 mL/kg.

ERG results for dark adaptation were obtained, then (during the same experiment) for the light adaptation phase. The reaction was started at least 30 minutes after the start of the light cycle for dark adaptation and the animals were left in a dark adaptation environment for at least 1 hour before recording.

At 18 and 66 hours after administration, mice were anesthetized with a mixture of ketamine and xylazine (100 mg/kg and 20mg/kg, respectively) and placed on a heating pad to maintain a stable core body temperature during the experiment. The pupil was dilated by dropping 5 microliters of mydriatic agent solution (0.5% tropicamide) into the noted intraocular lens. A mouse corneal unipolar lens contact electrode (Mayo corporation, Inazawa, aici, japan) was placed on the cornea, and a 12mm subcutaneous low profile needle-shaped reference electrode (GrassTelefactor, WWarwick, RI) was placed in the middle of the eyeball. The needle-shaped grounding electrode is arranged at the tail part. Using espion E²(DiagnosysLLC, Littleton, MA) ERG recording system data acquisition was obtained with a colorful hemisphere Ganzfeld (Ganzfeld) stimulator. The response function of the full dark adaptive intensity is a short one from 0.0001cd.s/m ²To 333cd.s/m²14 intensities of white flash stimuli over the range. Then, the full-scale adaptive intensity response function is passed through a short time from 0.33cd.s/m²To 333cd.s/m²Range of 9 intensities of white flash stimuli. The obtained reaction was analyzed off-line. The intensity response function determination was done by fitting the data to a Sigmoid function (NakaKI, RushtonWA,1966; NakaKI, RushtonWA, 1967). The ERG responses are listed in tables 17 and 18 below.

TABLE 17

Dark adapted ERG response

^*Ensemble averaging;

^**comparison with 4 hours Carrier

Watch 18

Photopic ERG response data

^*Ensemble averaging;

^**comparison with 4 hours Carrier

Dark-adapted ERG was inhibited at a dose rate of two doses (0.5 and 1mg/kg) of the compound 4 hours after administration. Both doses improved the maximum photopic ERG response. At 25 hours after application, dark adaptation ERG increased significantly at 25 hours with residual levels within <20% of vehicle treated animals. The residual response of the photopic ERG was enhanced.

The related examples describe the determination of the effect of alkyne-derived compounds as visual cycle modulators at the level of ERG response in the mouse eyeball following oral administration of the compounds to animals. The experimental procedure was the same as that described in the previous examples, except that 12-16 weeks old BALB/c mice (body weight 16-25 g) were used. The ERG responses are listed in tables 19 and 20 below.

Watch 19

Dark-adapted ERG response data

^*Ensemble averaging;

^**comparison with 4 hours Carrier

Watch 20

Photopic ERG response data

^*Ensemble averaging;

^**comparison with 4 hours Carrier

Administration resulted in a significant and dose-dependent (53% -96%) inhibition of the ERG dark adaptation intensity response function at three doses. This is accompanied by photopic adaptation V_maxRelatively small of (<15%) was added.

Example 199

Effect of alkynyl phenyl derivative compound on lipofuscin fluorescent substance reduction

This example describes the ability of alkynyl phenyl derivative compounds to reduce the level of A2E present in mouse retinas and to prevent the formation of A2E.

Abca4-null (abca4-/-) mutant mice (see, e.g., Wengetal, Cell98:13-23 (1999)) have increased accumulation of lipofuscin fluorescence in the eyeball, such as A2E (see, e.g., Karantal, Proc. Natl. Acad. Sci. USA102:4164-69 (2005)). Abca 4-/-mice about two months old are given daily oral gavage of compound (1mg/kg) or vehicle for three months. mice are sacrificed after three months of treatment.retinas and RPE are extracted for A2E analysis.

Similar experiments were performed with adult balb/c mice (10 months of age). Test mice were treated with 1 mg/kg/day of compound for three months and control mice were treated with vehicle.

Example 200

Effect of alkynyl phenyl derivative Compounds on retinoid Nuclear receptor Activity

Retinoid nuclear receptor activity is associated with transduction of non-ocular physiological, pharmacological and toxicological retinoid signals that affect tissue and organ growth, development, differentiation and homeostasis.

The effects of compound 4, compound 28 and compound 29, and Retinoic Acid Receptor (RAR) agonists (E-4- [2- (5,6,7, 8-tetrahydro-5, 5,8, 8-tetramethyl-2-naphthylene) -1-propenyl-receptor (RAR) were studied at the Retinoic Acid Receptor (RAR) and Retinoid X Receptor (RXR) in general accordance with Achkar et al (Proc. Natl. Acad. Sci. USA93:4879-84(1996))]Benzoic acid) (TTNPB) and all-trans retinoic acid (at-RA) as agonists of RAR and Retinoid X Receptors (RXR). The results of these analyses are shown in Table 6. Compound 4-HCl, compound 28-HCl and compound 29-HCl each showed no significant effect on retinoid nuclear receptors (RAR and RXR) at levels up to 10. mu.M. In contrast, TTNPB and at-RA activated RXR as expected_α、RAR_α、RAR_βAnd RAR_γReceptors (table 21).

TABLE 21

N/D = no activity detected; N/A = not applicable

When ranges are used herein for physical properties such as molecular weight or chemical properties such as chemical formulas, all combinations and subcombinations of the ranges and specific embodiments are intended to be included herein.

Various embodiments provided herein may be combined to provide further embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and/or listed in the application data sheet, are incorporated herein by reference, in their entirety.

From the foregoing, it will be appreciated that, although specific embodiments have been described herein for purposes of illustration, various modifications may be made. Those skilled in the art will understand, or be able to ascertain using no more than routine experimentation, equivalents to the specific embodiments described herein. Such equivalents are intended to be encompassed by the following claims. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the disclosure.

While a preferred embodiment of the invention has been shown and described herein, it will be understood by those skilled in the art that this embodiment is provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that many variations of the embodiments of the invention described herein may be used to practice the invention. It is intended that the following claims define the scope of the invention and that methods and structures within these claims and their equivalents be covered thereby.

Claims

1. A compound represented by the general formula (E):

or a pharmaceutically acceptable salt thereof, wherein:

m is 0 or 1;

R²¹and R²²Each independently is hydrogen or C₁～C₅An alkyl group;

R³and R⁴Each is independentThe site is hydrogen;

R⁵is C₁～C₁₃Alkyl radical, C₆～C₁₈Aryl, or C₃～C₁₀A carbocyclic group;

R¹²and R¹³Are the same or different and are independently hydrogen, -CO₂R⁹or-C (═ O) R⁹；

R⁹Is C₁～C₈An alkyl group;

each R is¹⁴Are the same or different and are independently C₁～C₅Alkyl, halogen, OR-OR⁶(ii) a And

each R is⁶Are the same or different and are independently hydrogen or C₁～C₅An alkyl group.

2. A compound represented by the general formula (F):

or a pharmaceutically acceptable salt thereof, wherein:

m is 0 or 1;

R²¹and R²²Each independently is hydrogen or C₁～C₅An alkyl group;

R³and R⁴Each independently is hydrogen;

R⁹Is C₁～C₈An alkyl group;

each R is¹⁴Are the same or different and are independently C₁～C₅Alkyl, halogen OR-OR⁶；

R¹⁶、R¹⁷And R¹⁸Each of which is the same or different and is independently hydrogen, C₁～C₈Alkyl, -OR⁶、C₃～C₁₀Carbocyclic radical or C₆～C₁₈An aryl group; and

3. The compound of claim 2, wherein R¹²And R¹³Each is hydrogen.

4. The compound of claim 3, wherein m is 0.

5. The compound of claim 4, wherein R ¹⁶、R¹⁷And R¹⁸Each independently is hydrogen, C₁～C₈Alkyl OR-OR⁶Wherein each R is⁶Independently is hydrogen or C₁～C₅An alkyl group; or wherein R is¹⁶、R¹⁷And R¹⁸Each independently is hydrogen, C₁～C₈Alkyl or C₆～C₁₈And (4) an aryl group.

6. The compound of claim 1, wherein R⁵Is C₃～C₁₀A carbocyclic group.

7. A compound represented by the general formula (G):

or a pharmaceutically acceptable salt thereof, wherein:

m is 0 or 1;

p is 1, 2, 3, 4 or 5;

q is 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9;

R²¹and R²²Each of which is the same or different and is independently hydrogen or C₁～C₅An alkyl group;

R³and R⁴Each independently is hydrogen;

R¹²and R¹³The same or different and independentlyIs hydrogen, -CO₂R⁹or-C (═ O) R⁹；

R⁹Is C₁～C₈An alkyl group;

Each R is¹⁹Are the same or different and are independently C₁～C₅Alkyl or OH; and

8. The compound of claim 7, wherein R¹²And R¹³Each is hydrogen.

9. The compound of claim 8, wherein m is 0.

10. The compound of claim 9, wherein q is 0 or 1.

11. The compound of claim 1, wherein m is 0 and R is¹²And R¹³Each is hydrogen.

12. The compound of claim 1, wherein R ⁵Is C₆～C₁₈And (4) an aryl group.

13. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of any one of claims 1 to 12.

14. Use of a compound of any one of claims 1 to 12 or a pharmaceutical composition of claim 13 in the manufacture of a medicament for modulating the amount of a chromophore in the retinoid circulation or for treating an ophthalmic disease or disorder in an individual.

15. The use of claim 14, wherein the medicament causes a reduction in lipofuscin pigment accumulated in the eye of the subject.

16. The use according to claim 15, wherein the lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).