PHENOXY DERIVATIVES AS

SPHINGOSINE 1 -PHOSPHATE (S1 P) RECEPTOR MODULATORS

By inventors: Wenkui K. Fang, Evelyn G. Corpuz and Ken Chow

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent Application No. 61/913,076 filed December 06, 2013, the entire disclosure of which is incorporated herein by this specific reference.

FIELD OF THE INVENTION

The present invention relates to phenoxy derivatives, processes for preparing them, pharmaceutical compositions containing them and their use as pharmaceuticals, as modulators of the sphingosine-1 -phosphate (S1 P) receptors. The invention relates specifically to the use of these compounds and their pharmaceutical compositions to treat disorders associated with S1 P receptor modulation.

BACKGROUND OF THE INVENTION

S1 P is stored in relatively high concentrations in human platelets, which lack the enzymes responsible for its catabolism, and it is released into the blood stream upon activation of physiological stimuli, such as growth factors, cytokines, and receptor agonists and antigens. It may also have a critical role in platelet aggregation and thrombosis and could aggravate cardiovascular diseases. On the other hand the relatively high concentration of the metabolite in high-density lipoproteins (HDL) may have beneficial implications for atherogenesis. For example, there are recent suggestions that S1 P, together with other lysolipids such as sphingosylphosphorylcholine and lysosulfatide, are responsible for the beneficial clinical effects of HDL by stimulating the production of the potent antiatherogenic signaling molecule nitric oxide by the vascular endothelium. In addition, like lysophosphatidic acid, it is a marker for certain types of cancer, and there is evidence that its role in cell division or proliferation may have an influence on the development of cancers. These are currently topics that are attracting great interest amongst medical researchers, and the potential for therapeutic intervention in S1 P metabolism is under active investigation.

SUMMARY OF THE INVENTION

A group of novel phenoxy derivatives, which are potent and selective S1 P modulators, has been discovered. As such, the compounds described herein are useful in treating a wide variety of disorders associated with modulation of the S1 P receptors. The term "modulator" as used herein, includes but is not limited to: receptor agonist, antagonist, inverse agonist, inverse antagonist, partial agonist, partial antagonist.

This invention describes compounds of Formula I, which have S1 P receptor biological activity. The compounds in accordance with the present invention are thus of use in medicine, for example, in the treatment of humans with diseases and conditions that are alleviated by S1 P modulation.

In one aspect, the invention provides a compound represented by Formula I:

Formula I

wherein:

R ¹ is H or F;

R ² is H or F;

R ³ is H or F;

R ⁴ is H, F, CI or -OCHs;

R ⁵ is F or -CH ₃ ; R ⁶ is -P(0)(OH) ₂ or -COOH; and

c is 0 or 1 ;

or an enantiomer, diastereomer or tautomer thereof;

or a pharmaceutically acceptable salt of any of the foregoing;

provided that at least one of R ¹ , R ² , R ³ and R ⁴ is not H.

Some compounds of the invention include:

3-[(4-{[5-(3,4-dimethylphenyl)-4-(2-fluoro-3-methoxypheny l)pentyl]oxy}benzyl) amino]propanoic acid;

{3-[(4-{[5-(3,4-dimethylphenyl)-4-(2-fluoro-3-methoxyphen yl)pentyl]oxy}

benzyl)amino]propyl}phosphonic acid;

3-[(4-{[5-(3,4-dimethylphenyl)-4-(3-fluoro-2-methylphenyl)pe ntyl]oxy}benzyl) amino]propanoic acid;

{3-[(4-{[5-(3,4-dimethylphenyl)-4-(3-fluoro-2-methylphenyl)p entyl]oxy}

benzyl)amino]propyl}phosphonic acid;

3-[(4-{[4-(2,6-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl] oxy}benzyl)amino]propanoic acid; {3-[(4-{[4-(2,6-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl ]oxy}benzyl)

amino]propyl}phosphonic acid;

3-[(4-{[4-(3-chloro-2-fluorophenyl)-5-(3,4-dimethylphenyl)pe ntyl]oxy}benzyl) amino]propanoic acid;

{3-[(4-{[4-(3-chloro-2-fluorophenyl)-5-(3,4-dimethylphenyl)p entyl]oxy}benzyl)

amino]propyl}phosphonic acid;

3-[(4-{[4-(2,5-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl] oxy}benzyl)amino]propanoic acid; {3-[(4-{[4-(2,5-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl ]oxy}benzyl)

amino]propyl}phosphonic acid;

3-[(4-{[4-(2,4-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl] oxy}benzyl)amino]propanoic acid; and

{3-[(4-{[4-(2,4-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl ]oxy}benzyl)

amino]propyl}phosphonic acid. The term "pharmaceutically acceptable salts" refers to salts or complexes that retain the desired biological activity of the above identified compounds and exhibit minimal or no undesired toxicological effects. The "pharmaceutically acceptable salts" according to the invention include therapeutically active, non-toxic base or acid salt forms, which the compounds of Formula I are able to form.

The acid addition salt form of a compound of Formula I that occurs in its free form as a base can be obtained by treating the free base with an appropriate acid such as an inorganic acid, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; or an organic acid such as for example, acetic acid, hydroxyacetic acid, propanoic acid, lactic acid, pyruvic acid, malonic acid, fumaric acid, maleic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, citric acid, methylsulfonic acid, ethanesulfonic acid, benzenesulfonic acid, formic and the like (Handbook of Pharmaceutical Salts, P. Heinrich Stahl & Camille G. Wermuth (Eds), Verlag Helvetica Chimica Acta, Zurich, 2002, 329-345). The base addition salt form of a compound of Formula I that occurs in its acid form can be obtained by treating the acid with an appropriate base such as an inorganic base, for example, sodium hydroxide, magnesium hydroxide, potassium hydroxide, calcium hydroxide, ammonia and the like; or an organic base such as for example, L-Arginine, ethanolamine, betaine, benzathine, morpholine and the like (Handbook of Pharmaceutical Salts, P. Heinrich Stahl & Camille G. Wermuth (Eds), Verlag Helvetica Chimica Acta, Zurich, 2002, 329-345).

Compounds of Formula I and their salts can be in the form of a solvate, which is included within the scope of the present invention. Such solvates include, for example, hydrates, alcoholates and the like.

With respect to the present invention, reference to a compound or compounds is intended to encompass that compound in each of its possible isomeric forms and mixtures thereof unless the particular isomeric form is referred to specifically. Compounds according to the present invention may exist in different polymorphic forms. Although not explicitly indicated in the above formula, such forms are intended to be included within the scope of the present invention.

The compounds of the invention are indicated for use in treating or preventing conditions in which there is likely to be a component involving the S1 P receptors.

In another embodiment, there are provided pharmaceutical compositions including at least one compound of the invention in a pharmaceutically acceptable carrier.

In a further embodiment of the invention, there are provided methods for treating disorders associated with modulation of S1 P receptors. Such methods can be performed, for example, by administering to a subject in need thereof a pharmaceutical composition containing a therapeutically effective amount of at least one compound of the invention.

Compounds of the invention are useful for the treatment of mammals, including humans, with a range of conditions and diseases that are alleviated by S1 P modulation.

Therapeutic utilities of S1 P modulators include diseases, such as but not limited to: Ocular Diseases: wet and dry age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity, retinal edema, dry eye, geographic atrophy, glaucomatous optic neuropathy, chorioretinopathy, hypertensive retinopathy, ocular ischemic syndrome, prevention of inflammation-induced fibrosis in the back of the eye, various ocular inflammatory diseases including uveitis, scleritis, keratitis, and retinal vasculitis; Systemic vascular barrier related diseases: various inflammatory diseases, including acute lung injury, its prevention, sepsis, tumor metastasis, atherosclerosis, pulmonary edemas, and ventilation-induced lung injury;

Autoimmune diseases and immnuosuppression: rheumatoid arthritis, Crohn's disease, Graves' disease, inflammatory bowel disease, multiple sclerosis, Myasthenia gravis, psoriasis, ulcerative colitis, antoimmune uveitis, renal ischemia/perfusion injury, contact hypersensitivity, atopic dermatitis, and organ transplantation; Allergies and other inflammatory diseases: urticaria, bronchial asthma, and other airway inflammations including pulmonary emphysema and chronic obstructive pulmonary diseases;

Cardiac functions: bradycardia, congestional heart failure, cardiac arrhythmia, prevention and treatment of atherosclerosis, and ischemia/reperfusion injury;

Wound Healing: scar-free healing of wounds from cosmetic skin surgery, ocular surgery, Gl surgery, general surgery, oral injuries, various mechanical, heat and burn injuries, prevention and treatment of photoaging and skin ageing, and prevention of radiation-induced injuries; Bone formation: treatment of osteoporosis and various bone fractures including hip and ankles;

Anti-nociceptive activity: visceral pain, pain associated with diabetic neuropathy, rheumatoid arthritis, chronic knee and joint pain, tendonitis, osteoarthritis, neuropathic pains; Anti-fibrosis: ocular, cardiac, hepatic and pulmonary fibrosis, proliferative vitreoretinopathy, cicatricial pemphigoid, surgically induced fibrosis in cornea, conjunctiva and tenon;

Pains and anti-inflammation: acute pain, flare-up of chronic pain, musculo-skeletal pains, visceral pain, pain associated with diabetic neuropathy, rheumatoid arthritis, chronic knee and joint pain, tendonitis, osteoarthritis, bursitis, neuropathic pains;

CNS neuronal injuries: Alzheimer's disease, age-related neuronal injuries;

Organ transplants: renal, corneal, cardiac and adipose tissue transplants.

In still another embodiment of the invention, there are provided methods for treating disorders associated with modulation of sphingosine-1 -phosphate receptors. Such methods can be performed, for example, by administering to a subject in need thereof a therapeutically effective amount of at least one compound of the invention, or any combination thereof, or pharmaceutically acceptable salts, hydrates, solvates, crystal forms and individual isomers, enantiomers, and diastereomers thereof.

The present invention concerns the use of a compound of Formula I or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of:

Ocular Diseases: wet and dry age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity, retinal edema, dry eye, geographic atrophy, glaucomatous optic neuropathy, chorioretinopathy, hypertensive retinopathy, ocular ischemic syndrome, prevention of inflammation-induced fibrosis in the back of the eye, various ocular inflammatory diseases including uveitis, scleritis, keratitis, and retinal vasculitis;

Systemic vascular barrier related diseases: various inflammatory diseases, including acute lung injury, its prevention, sepsis, tumor metastasis, atherosclerosis, pulmonary edemas, and ventilation-induced lung injury;

Autoimmune diseases and immnuosuppression: rheumatoid arthritis, Crohn's disease, Graves' disease, inflammatory bowel disease, multiple sclerosis, Myasthenia gravis, psoriasis, ulcerative colitis, antoimmune uveitis, renal ischemia/perfusion injury, contact hypersensitivity, atopic dermatitis, and organ transplantation;

Allergies and other inflammatory diseases: urticaria, bronchial asthma, and other airway inflammations including pulmonary emphysema and chronic obstructive pulmonary diseases; Cardiac functions: bradycardia, congestional heart failure, cardiac arrhythmia, prevention and treatment of atherosclerosis, and ischemia/reperfusion injury;

Wound Healing: scar-free healing of wounds from cosmetic skin surgery, ocular surgery, Gl surgery, general surgery, oral injuries, various mechanical, heat and burn injuries, prevention and treatment of photoaging and skin ageing, and prevention of radiation-induced injuries;

Bone formation: treatment of osteoporosis and various bone fractures including hip and ankles; Anti-nociceptive activity: visceral pain, pain associated with diabetic neuropathy, rheumatoid arthritis, chronic knee and joint pain, tendonitis, osteoarthritis, neuropathic pains;

Anti-fibrosis: ocular, cardiac, hepatic and pulmonary fibrosis, proliferative vitreoretinopathy, cicatricial pemphigoid, surgically induced fibrosis in cornea, conjunctiva and tenon;

Pains and anti-inflammation: acute pain, flare-up of chronic pain, musculo-skeletal pains, visceral pain, pain associated with diabetic neuropathy, rheumatoid arthritis, chronic knee and joint pain, tendonitis, osteoarthritis, bursitis, neuropathic pains;

CNS neuronal injuries: Alzheimer's disease, age-related neuronal injuries; Organ transplants: renal, corneal, cardiac and adipose tissue transplants.

The actual amount of the compound to be administered in any given case will be determined by a physician taking into account the relevant circumstances, such as the severity of the condition, the age and weight of the patient, the patient's general physical condition, the cause of the condition, and the route of administration. The patient will be administered the compound orally in any acceptable form, such as a tablet, liquid, capsule, powder and the like, or other routes may be desirable or necessary, particularly if the patient suffers from nausea. Such other routes may include, without exception, transdermal, parenteral, subcutaneous, intranasal, via an implant stent, intrathecal, intravitreal, topical to the eye, back to the eye, intramuscular, intravenous, and intrarectal modes of delivery. Additionally, the formulations may be designed to delay release of the active compound over a given period of time, or to carefully control the amount of drug released at a given time during the course of therapy.

In another embodiment of the invention, there are provided pharmaceutical compositions including at least one compound of the invention in a pharmaceutically acceptable carrier thereof. The phrase "pharmaceutically acceptable" means the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. Pharmaceutical compositions of the present invention can be used in the form of a solid, a solution, an emulsion, a dispersion, a patch, a micelle, a liposome, and the like, wherein the resulting composition contains one or more compounds of the present invention, as an active ingredient, in admixture with an organic or inorganic carrier or excipient suitable for enteral or parenteral applications. Invention compounds may be combined, for example, with the usual non-toxic, pharmaceutically acceptable carriers for tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, and any other form suitable for use. The carriers which can be used include glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea, medium chain length triglycerides, dextrans, and other carriers suitable for use in manufacturing preparations, in solid, semisolid, or liquid form. In addition auxiliary, stabilizing, thickening and coloring agents and perfumes may be used. Invention compounds are included in the pharmaceutical composition in an amount sufficient to produce the desired effect upon the process or disease condition. Pharmaceutical compositions containing invention compounds may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of a sweetening agent such as sucrose, lactose, or saccharin, flavoring agents such as peppermint, oil of wintergreen or cherry, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets containing invention compounds in admixture with non-toxic pharmaceutically acceptable excipients may also be manufactured by known methods. The excipients used may be, for example, (1 ) inert diluents such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating and disintegrating agents such as corn starch, potato starch or alginic acid; (3) binding agents such as gum tragacanth, corn starch, gelatin or acacia, and (4) lubricating agents such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed.

In some cases, formulations for oral use may be in the form of hard gelatin capsules wherein the invention compounds are mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatin capsules wherein the invention compounds are mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil.

The pharmaceutical compositions may be in the form of a sterile injectable suspension. This suspension may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides, fatty acids (including oleic acid), naturally occurring vegetable oils like sesame oil, coconut oil, peanut oil, cottonseed oil, etc., or synthetic fatty vehicles like ethyl oleate or the like. Buffers, preservatives, antioxidants, and the like can be incorporated as required.

Pharmaceutical compositions containing invention compounds may be in a form suitable for topical use, for example, as oily suspensions, as solutions or suspensions in aqueous liquids or nonaqueous liquids, or as oil-in-water or water-in-oil liquid emulsions. Pharmaceutical compositions may be prepared by combining a therapeutically effective amount of at least one compound according to the present invention, or a pharmaceutically acceptable salt thereof, as an active ingredient with conventional ophthalmically acceptable pharmaceutical excipients and by preparation of unit dosage suitable for topical ocular use. The therapeutically efficient amount typically is between about 0.0001 and about 5% (w/v), preferably about 0.001 to about 2.0% (w/v) in liquid formulations.

For ophthalmic application, preferably solutions are prepared using a physiological saline solution as a major vehicle. The pH of such ophthalmic solutions should preferably be maintained between 4.5 and 8.0 with an appropriate buffer system, a neutral pH being preferred but not essential. The formulations may also contain conventional pharmaceutically acceptable preservatives, stabilizers and surfactants. Preferred preservatives that may be used in the pharmaceutical compositions of the present invention include, but are not limited to, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetate and phenylmercuric nitrate. A preferred surfactant is, for example, Tween 80. Likewise, various preferred vehicles may be used in the ophthalmic preparations of the present invention. These vehicles include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropyl methyl cellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl cellulose cyclodextrin and purified water. Tonicity adjustors may be added as needed or convenient. They include, but are not limited to, salts, particularly sodium chloride, potassium chloride, mannitol and glycerin, or any other suitable ophthalmically acceptable tonicity adjustor.

Various buffers and means for adjusting pH may be used so long as the resulting preparation is ophthalmically acceptable. Accordingly, buffers include acetate buffers, citrate buffers, phosphate buffers and borate buffers. Acids or bases may be used to adjust the pH of these formulations as needed.

In a similar manner an ophthalmically acceptable antioxidant for use in the present invention includes, but is not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene. Other excipient components which may be included in the ophthalmic preparations are chelating agents. The preferred chelating agent is edentate disodium, although other chelating agents may also be used in place of or in conjunction with it.

The ingredients are usually used in the following amounts: Ingredient Amount (% w/v)

active ingredient about 0.001 -5

preservative 0-0.10

vehicle 0-40 tonicity adjustor 0-10

buffer 0.01 -10

pH adjustor q .s. pH 4.5-7.8

antioxidant as needed

surfactant as needed

purified water to make 100%

The actual dose of the active compounds of the present invention depends on the specific compound, and on the condition to be treated; the selection of the appropriate dose is well within the knowledge of the skilled artisan.

The ophthalmic formulations of the present invention are conveniently packaged in forms suitable for metered application, such as in containers equipped with a dropper, to facilitate application to the eye. Containers suitable for drop wise application are usually made of suitable inert, non-toxic plastic material, and generally contain between about 0.5 and about 15 ml solution. One package may contain one or more unit doses. Especially preservative-free solutions are often formulated in non-resalable containers containing up to about ten, preferably up to about five units doses, where a typical unit dose is from one to about 8 drops, preferably one to about 3 drops. The volume of one drop usually is about 20-35 microliters.

Invention compounds may also be administered in the form of suppositories for rectal administration of the drug. These compositions may be prepared by mixing the invention compounds with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters of polyethylene glycols, which are solid at ordinary temperatures, but liquefy and/or dissolve in the rectal cavity to release the drug.

Since individual subjects may present a wide variation in severity of symptoms and each drug has its unique therapeutic characteristics, the precise mode of administration and dosage employed for each subject is left to the discretion of the practitioner.

The compounds and pharmaceutical compositions described herein are useful as medicaments in mammals, including humans, for treatment of diseases and/or alleviations of conditions which are responsive to treatment by agonists or functional antagonists of sphingosine-1 -phosphate receptors. Thus, in further embodiments of the invention, there are provided methods for treating a disorder associated with modulation of sphingosine-1 - phosphate receptors. Such methods can be performed, for example, by administering to a subject in need thereof a pharmaceutical composition containing a therapeutically effective amount of at least one invention compound. As used herein, the term "therapeutically effective amount" means the amount of the pharmaceutical composition that will elicit the biological or medical response of a subject in need thereof that is being sought by the researcher, veterinarian, medical doctor or other clinician. In some embodiments, the subject in need thereof is a mammal. In some embodiments, the mammal is human. The present invention also concerns processes for preparing the compounds of

Formula I. The compounds of Formula I according to the invention can be prepared analogously to conventional methods as understood by the person skilled in the art of synthetic organic chemistry. The synthetic schemes set forth below illustrate how compounds according to the invention can be made. The definitions of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , c and R ⁶ in the schemes are as defined in Formula I. Those skilled in the art will be able to routinely modify and/or adapt the following schemes to synthesize any compounds of the invention covered by Formula I.

Scheme 1a

Scheme 1c

Scheme 1 d

Description of Scheme 1 a A suitably substituted acid chloride and a suitably substituted arene were mixed and cooled to -78°C; AICI ₃ (1 .5 eq) was added, the resulting reaction mixture was stirred at this temperature for 60 minutes, and then quenched with concentrated HCI. The reaction mixture thus created was diluted with ethyl acetate and washed with brine, then concentrated. Flash chromatography gave the desired ketone. Description of Scheme 1 b

In another reaction vessel, a suitably substituted hydroxybenzoic acid ester was mixed with 1 ,3-dibromopropane and cooled to 0°C. NaH (60% suspension in mineral oil, 1 .5 eq) was slowly added, the resulting reaction mixture was stirred at 95°C for 4 hours, then concentrated. The reaction mixture was then diluted with ethyl acetate, and washed with saturated ammonium chloride and brine, then dried (Na ₂ S0 ₄ ) and concentrated. Flash chromatography gave the desired bromopropylloxybenzoic acid ester.

Description of Scheme 1 c The above-prepared ketone (Scheme 1a) was added to LDA (prepared fresh from n-

BuLi and diisopropylamine in THF) at -78°C, then warmed to 0°C and stirred for 60 minutes. The reaction mixture was again cooled to -78°C, the bromopropylloxybenzoic acid ester prepared above (Scheme 1 b) was added, the resultant mixture was allowed to warm to room temperature and stirred for 4 hours, then heated to 95°C and stirred for another 14 hours. After dilution with ethyl acetate, the resulting reaction mixture was washed with saturated ammonium chloride and brine, then dried (Na ₂ S0 ₄ ) and concentrated. This alkylated ketone was then dissolved in methanol and cooled to 0°C, and sodium borohydride (1 .0 eq) was slowly added. The reaction mixture was stirred at 0°C for 10 minutes, then diluted with ethyl acetate and the resultant organic phase was washed with saturated ammonium chloride and brine, then dried and concentrated. Flash chromatography gave the desired compound which was dissolved in dichloromethane and cooled to 0°C. Trifluoroacetic acid (2 eq) was added, followed by the addition of triethylsilane. The resulting reaction mixture was allowed to warm to room temperature and stirred for 4 hours, then concentrated. This intermediate ester was dissolved in ether, then cooled to 0°C. LAH (1 .5 eq) was added, the resulting reaction mixture was stirred at the same temperature for 60 minutes, then diluted HCI was added. The organic phase was separated and washed with brine, then dried (Na ₂ S0 ₄ ) and concentrated. Flash chromatography gave the desired alcohol.

Description of Scheme 1 d

The alcohol prepared above (Scheme 1c) was mixed with NMO (26 eq), molecular sieve (500mg) in AcCN:DCM (1 :5). A catalytic amount of TPAP (35mg) was added. The resulting reaction mixture was stirred at RT for 1 hour and then evaporated to dryness. The desired aldehyde compound was purified by MPLC using 0-10% ethyl acetate in hexanes. Procedure B

A sample of the above-prepared aldehyde (see Description of Scheme 1 d) was dissolved in minimum THF. Methanol (5ml_) was then added followed by 5 drops of HOAc. A suitable intermediate bearing an amine and a carboxylate group (e.g., β-alanine) was added, and the resulting mixture was stirred at room temperature for 30 minutes. NaCNBH ₃ was dissolved in 0.5 mL MeOH and added to the reaction. The resulting mixture was stirred at room temperature for 3 hours, then quenched with water and concentrated on the rotary evaporator. The crude mixture was purified by MPLC to give the final product. Procedure C

A mixture of a suitable intermediate bearing an amine and a phosphonic acid group (e.g., 3-aminopropylphosphonic acid) and Bu ₄ NOH in methanol was stirred at 30 °C for a few minutes. The above prepared aldehyde (see Description of Scheme 1d) dissolved in a minimum amount of THF was added. The reaction mixture was stirred at 50 °C for 3 hours, then NaCNBH ₃ dissolved in minimum amount of MeOH was added. The resulting mixture was stirred at 50 °C for 3 hours, cooled to room temperature and quenched with water, and then concentrated on the rotary evaporator and purified by MPLC to give the final product.

The following abbreviations are used herein:

AcCN acetonitrile

DCM dichloromethane

MeOH methanol

CD ₃ OD deuterated methanol

RT or rt room temperature

AcOH acetic acid

MPLC Medium Pressure Liquid Chromatography

NMO 4-methylmorpholine N-oxide

TPAP tetrapropylammonium perruthenate

NaCNBHs sodium cyanoborohydride

AICI3 aluminum chloride

HCI hydrochloric acid NaH sodium hydrate

Na ₂ S0 sodium sulfate

LDA lithium di-isopropylamide

n-BuLi n-butyllithium

THF tetrahydrofuran

LAH lithium aluminum hydride

TMS tetramethylsilane

DRAWINGS

Figure 1 shows the structure of Formula I.

DETAILED DESCRIPTION OF THE INVENTION

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention claimed. As used herein, the use of the singular includes the plural unless specifically stated otherwise.

It will be readily apparent to those skilled in the art that some of the compounds of the invention may contain one or more asymmetric centers, such that the compounds may exist in enantiomeric as well as in diastereomeric forms. Unless it is specifically noted otherwise, the scope of the present invention includes all enantiomers, diastereomers and racemic mixtures. Some of the compounds of the invention may form salts with pharmaceutically acceptable acids or bases, and such pharmaceutically acceptable salts of the compounds described herein are also within the scope of the invention.

The present invention includes all pharmaceutically acceptable isotopically enriched compounds. Any compound of the invention may contain one or more isotopic atoms enriched or different than the natural ratio such as deuterium ² H (or D) in place of protium ¹ H (or H) or use of ¹³ C-enriched material in place of ¹² C and the like. Similar substitutions can be employed for N, O and S. The use of isotopes may assist in analytical as well as therapeutic aspects of the invention. For example, use of deuterium may increase the in vivo half-life by altering the metabolism (rate) of the compounds of the invention. These compounds can be prepared in accord with the preparations described by use of isotopically enriched reagents.

The following examples are for illustrative purposes only and are not intended, nor should they be construed as limiting the invention in any manner. Those skilled in the art will appreciate that variations and modifications of the following examples can be made without exceeding the spirit or scope of the invention.

As will be evident to those skilled in the art, individual diastereomeric forms can be obtained by separation of mixtures thereof in conventional manner; chromatographic separation may be employed.

The following are non-limiting embodiments of the invention.

In embodiment (1 ), there is provided a compound having Formula I:

Formula I

wherein:

R is H or F

R ² is H or F;

R ³ is H or F;

R ⁴ is H, F, CI or -OCHs;

R ⁵ is F or -CH ₃ ;

R ⁶ is -P(0)(OH) ₂ or -COOH; and c is 0 or 1 ;

or an enantiomer, diastereomer or tautomer thereof;

or a pharmaceutically acceptable salt of any of the foregoing;

provided that at least one of R ¹ , R ² , R ³ and R ⁴ is not H.

In embodiment (2), there is provide a compound of embodiment (1 ), wherein the compound is selected from:

3-[(4-{[5-(3,4-dimethylphenyl)-4-(2-fluoro-3-methoxyphenyl)p entyl]oxy}benzyl) amino]propanoic acid;

{3-[(4-{[5-(3,4-dimethylphenyl)-4-(2-fluoro-3-methoxyphen yl)pentyl]oxy}

benzyl)amino]propyl}phosphonic acid;

3-[(4-{[5-(3,4-dimethylphenyl)-4-(3-fluoro-2-methylphenyl)pe ntyl]oxy}benzyl) aminojpropanoic acid;

{3-[(4-{[5-(3,4-dimethylphenyl)-4-(3-fluoro-2-methylphenyl)p entyl]oxy}

benzyl)amino]propyl}phosphonic acid;

3-[(4-{[4-(2,6-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl] oxy}benzyl)amino]propanoic acid; {3-[(4-{[4-(2,6-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl ]oxy}benzyl)

amino]propyl}phosphonic acid;

3-[(4-{[4-(3-chloro-2-fluorophenyl)-5-(3,4-dimethylphenyl)pe ntyl]oxy}benzyl) aminojpropanoic acid;

{3-[(4-{[4-(3-chloro-2-fluorophenyl)-5-(3,4-dimethylphenyl)p entyl]oxy}benzyl)

amino]propyl}phosphonic acid;

3-[(4-{[4-(2,5-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl] oxy}benzyl)amino]propanoic acid; {3-[(4-{[4-(2,5-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl ]oxy}benzyl)

amino]propyl}phosphonic acid;

3-[(4-{[4-(2,4-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl] oxy}benzyl)amino]propanoic acid; and

{3-[(4-{[4-(2,4-difluorophenyl)-5-(3,4-dimethylphenyl)pentyl ]oxy}benzyl)

amino]propyl}phosphonic acid. In embodiment (3), there is provided a pharmaceutical composition comprising as active ingredient a therapeutically effective amount of a compound according to embodiment (1 ) or (2), and a pharmaceutically acceptable diluent, excipient or carrier.

In embodiment (4) there is provided a method of treating a condition associated with sphingosine-1 -phosphate receptor modulation, the method comprising administering to a mammal in need thereof a pharmaceutical composition comprising a therapeutically effective amount of at least one compound according any one of embodiments (1 ), (2) or (3).

In embodiment (5), there is provided a method of embodiment (4) wherein the condition is an ocular disease, preferably selected from wet and dry age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity, retinal edema, dry eye, geographic atrophy, glaucomatous optic neuropathy, chorioretinopathy, hypertensive retinopathy, ocular ischemic syndrome, prevention of inflammation-induced fibrosis in the back of the eye, ocular inflammatory diseases including uveitis, scleritis, keratitis, and retinal vasculitis.

In embodiment (6), there is provided a method of embodiment (4) wherein condition is a systemic vascular barrier related disease, preferably selected from inflammatory diseases, including acute lung injury, its prevention, sepsis, tumor metastasis, atherosclerosis, pulmonary edemas, and ventilation-induced lung injury.

In embodiment (7), there is provided a method of embodiment (4) wherein the condition is an autoimmune or immunosuppressive disease or disorder, wherein the disease or disorder is preferably selected from rheumatoid arthritis, Crohn's disease, Graves' disease, inflammatory bowel disease, multiple sclerosis, Myasthenia gravis, psoriasis, ulcerative colitis, antoimmune uveitis, renal ischemia/perfusion injury, contact hypersensitivity, atopic dermatitis, and organ transplantation.

In embodiment (8), there is provided a method of embodiment (4) wherein the condition is an allergy or other inflammatory condition, preferably selected from urticaria, bronchial asthma, and other airway inflammations (including pulmonary emphysema and chronic obstructive pulmonary diseases). In embodiment (9), there is provided a method of embodiment (4) wherein the condition is associated with cardiac function, preferably selected from bradycardia, congestional heart failure, cardiac arrhythmia, prevention and treatment of atherosclerosis, and ischemia/reperfusion injury. In embodiment (10), there is provided a method of embodiment (4) wherein the condition is wound healing, preferably selected from scar-free healing of wounds from cosmetic skin surgery, ocular surgery, gastrointestinal surgery, general surgery, oral injuries, various mechanical, heat and burn injuries, prevention and treatment of photoaging and skin ageing, and prevention of radiation-induced injuries. In embodiment (1 1 ), there is provided a method of embodiment (4) wherein the condition is associated with bone formation, preferably selected from osteoporosis and bone fractures.

In embodiment (12), there is provided a method of embodiment (4) wherein the condition is associated with anti-nociceptive activity, preferably selected from visceral pain, pain associated with diabetic neuropathy, rheumatoid arthritis, chronic knee and joint pain, tendonitis, osteoarthritis and neuropathic pain.

In embodiment (13), there is provided a method of embodiment (4) wherein the condition is anti-fibrosis, preferably selected from ocular, cardiac, hepatic and pulmonary fibrosis, proliferative vitreoretinopathy, cicatricial pemphigoid, surgically induced fibrosis in cornea, conjunctiva or tenon.

In embodiment (14), there is provided a method of embodiment (4) wherein the condition is pain or inflammation, preferably selected from acute pain, flare-up of chronic pain, musculo-skeletal pains, visceral pain, pain associated with diabetic neuropathy, rheumatoid arthritis, chronic knee and joint pain, tendonitis, osteoarthritis, bursitis and neuropathic pain. In embodiment (15), there is provided a method of embodiment (4) wherein the condition is associated with CNS neuronal injuries, preferably selected from Alzheimer's disease and age-related neuronal injuries. In embodiment (16), there is provided a method of embodiment (4) wherein the condition is associated with organ transplants, preferably selected from renal, corneal, cardiac and adipose tissue transplants.

Compound names were generated with ACDLabs version 12.5 and intermediates and reagent names used in the examples were generated with software such as Chem Bio Draw Ultra version 12.0 or Auto Norn 2000 from MDL ISIS Draw 2.5 SP1 .

In general, characterization of the compounds is performed according to the following methods: NMR spectra are recorded on 300 and/or 600 MHz Varian NMR spectrometer and acquired at room temperature; or at 60 MHz on a Varian T-60 spectrometer or at 300 MHz on a Varian Inova system. Chemical shifts are given in ppm referenced either to internal TMS or to the solvent signal.

All the reagents, solvents, catalysts for which the synthesis is not described are purchased from chemical vendors such as Sigma Aldrich, Fluka, Bio-Blocks, Combi-blocks, TCI, VWR, Lancaster, Oakwood, Trans World Chemical, Alfa, AscentScientific LLC, Fisher, Maybridge, Frontier, Matrix, Ukrorgsynth, Toronto, Ryan Scientific, SiliCycle, Anaspec, Syn Chem, Chem-lmpex, MIC-scientific, Ltd; however, some known intermediates were prepared according to published procedures. The compounds of the invention were purified by column chromatography (Auto-column) on a Teledyne-ISCO CombiFlash with a silica column, unless noted otherwise. Intermediates 1 through 6 (Table 1 ) and Compounds 1 through 12 (Table 2) were prepared according to the general procedures described above (See Schemes 1 a through 1 d). Table 2 also depicts the intermediate used in the preparation of Compounds 1 through 12. Those skilled in the art will be able to routinely modify and/or adapt the procedures to synthesize any compounds of the invention covered by Formula I.

Table 1

Interm. Name Ή NMR δ (ppm)

Structure

No.

1 4-{[5-(3,4-dimethylphenyl)-4-(2-fluoro-3- ^] H NMR (300 MHz, CDCI ₃ ) δ

methoxyphenyl)pentyl]oxy}benzaldehyde ppm 1 .50 - 1 .71 (m, 2 H) 1 .71 - H) 2.75 -2.89 (m, 21-1)3.16- 3.36 (m, 1 H)3.94 (t, =6.15Hz,

2 H) 6.70 -7.03 (m, 8 H) 7.80 (d, J=8.79 Hz, 2 H) 9.87 (s, 1 H)

6 4-{[4-(2,4-difluorophenyl)-5-(3,4- ^] H NMR (300 MHz, CDCI ₃ ) δ dimethylphenyl)pentyl]oxy}benzaldehyde ppm 1.59 - 1.73 (m, 2 H) 1.74 -

F 1.97 (m, 2 H)2.18 (br. s, 6 H)

2.84 (d, J=7.33 Hz, 2 H) 3.11 - 3.33 (m, 1 H)3.94 (t, J=6.15Hz, 2 H) 6.65 -6.86 (m, 4 H) 6.87 - 6.99 (m, 3 H) 7.03 -7.21 (m, 1 H) 7.80 (d, J=8.79 Hz, 2 H) 9.87

(s, 1 H)

Table 2

Biological Data

Compounds were synthesized and tested for S1 P1 activity using the GTP γ ³ ¾ binding assay. These compounds may be assessed for their ability to activate or block activation of the human S1 P1 receptor in cells stably expressing the S1 P1 receptor.

GTP Y ³⁵ S binding was measured in the medium containing (mM) HEPES 25, pH 7.4, MgCI ₂ 10, NaCI 100, dithitothreitol 0.5, digitonin 0.003%, 0.2 nM GTP y ³⁵ S, and 5 pg membrane protein in a volume of 150 μΙ. Test compounds were included in the concentration range from 0.08 to 5,000 nM unless indicated otherwise. Membranes were incubated with 100 μΜ 5'-adenylylimmidodiphosphate for 30 min, and subsequently with 10 μΜ GDP for 10 min on ice. Drug solutions and membrane were mixed, and then reactions were initiated by adding GTP Y ³⁵ S and continued for 30 min at 25 °C. Reaction mixtures were filtered over Whatman GF/B filters under vacuum, and washed three times with 3 mL of ice-cold buffer (HEPES 25, pH7.4, MgCl ₂ 10 and NaCI 100). Filters were dried and mixed with scintillant, and counted for ³⁵ S activity using a β-counter. Agonist-induced GTP y ³⁵ S binding was obtained by subtracting that in the absence of agonist. Binding data were analyzed using a non-linear regression method. In case of antagonist assay, the reaction mixture contained 10 nM S1 P in the presence of test antagonist at concentrations ranging from 0.08 to 5000 nM. Table 3 shows activity potency: S1 P1 receptor from GTP y ³⁵ S: nM, (EC ₅₀ ).

Table 3