AU2008206495A1 - 6-substituted indole-3-carboxylic acid amide compounds having Sphingosine-1-Phosphate (S1P) receptor antagonist biological activity - Google Patents

Description

WO 2008/089015 PCT/US2008/050695 1 6-SUBSTITUTED INDOLE-3-CARBOXYLIC ACID AMIDE COMPOUNDS HAVING SPHINGOSINE-1-PHOSPHATE (SiP) 5 RECEPTOR ANTAGONIST BIOLOGICAL ACTIVITY Inventors RICHARD L. BEARD HAIGING YUAN 10 CROSS REFERENCE This application claims the benefit of U.S. Provisional Application serial number 60/884,470, filed January 11, 2007 which is hereby incorporated by 15 reference in its entirety. BACKGROUND OF THE INVENTION 1. Field of the Invention 20 The present invention relates to derivatives and/or analogues of sphingosine and pharmaceutical compositions, including such derivatives and/or analogues, which are useful as drugs for the treatment of fungal infections, allergic diseases, immune disorders, etc. 25 2. Summary of the Art Sphingosine is a compound having the chemical structure shown in the general formula described below, in which Y' is hydrogen. It is known that various 30 sphingolipids, having sphingosine as a constituent, are widely distributed in the living body including on the surface of cell membranes of cells in the nervous system. H OH NH 2 I - 35 H- 3 C -(CH 2

)

12 -C =CH -CH -CH -CH 2 0-Y' WO 2008/089015 PCT/US2008/050695 2 H A sphingolipid is one of the lipids having important roles in the living body. A disease called lipidosis is caused by accumulation of a specified sphingolipid in the body. Sphingolipids present on cell membranes function to regulate cell 5 growth; participate in the development and differentiation of cells; function in nerves; are involved in the infection and malignancy of cells; etc. Many of the physiological roles of sphingolipids remain to be solved. Recently the possibility that ceramide, a derivative of sphingosine, has an important role in the mechanism of cell signal transduction has been indicated, and studies about 10 its effect on apoptosis and cell cycle have been reported. Sphingosine-1-phosphate is an important cellular metabolite, derived from ceramide that is synthesized de novo or as part of the sphingomeyeline cycle (in animals cells). It has also been found in insects, yeasts and plants. 15 The enzyme, ceramidase, acts upon ceramides to release sphingosine, which is phosphorylated by spingosine kinase, a ubiquitous enzyme in the cytosol and endoplasmic reticulum, to form sphingosine-1-phosphate. The reverse reaction can occur also by the action of sphingosine phosphatases, and the enzymes act 20 in concert to control the cellular concentrations of the metabolite, which concentrations are always low. In plasma, such concentration can reach 0.2 to 0.9 tM, and the metabolite is found in association with the lipoproteins, especially the HDL. It should also be noted that sphingosine- 1-phosphate formation is an essential step in the catabolism of sphingoid bases. 25 Like its precursors, sphingosine- 1-phosphate is a potent messenger molecule that perhaps uniquely operates both intra- and inter-cellularly, but with very different functions from ceramides and sphingosine. The balance between these various sphingolipid metabolites may be important for health. For example, 30 within the cell, sphingosine- 1-phosphate promotes cellular division (mitosis) as opposed to cell death (apoptosis), which it inhibits. Intracellularly, it also WO 2008/089015 PCT/US2008/050695 3 functions to regulate calcium mobilization and cell growth in response to a variety of extracellular stimuli. Current opinion appears to suggest that the balance between sphingosine-1-phosphate and ceramide and/or spingosine levels in cells is critical for their viability. In common with the 5 lysophospholipids, especially lysophosphatidic acid, with which it has some structural similarities, sphingosine- 1-phosphate exerts many of its extra-cellular effects through interaction with five specific G protein-coupled receptors on cell surfaces. These are important for the growth of new blood vessels, vascular maturation, cardiac development and immunity, and for directed cell movement. 10 Sphingosine- 1 phosphate 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 15 critical role in platelet aggregation and thrombosis and could aggravate cardiovascular disease. 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 sphingosine-1-phosphate, together with other lysolipids such as 20 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 25 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 sphingosine-1-phosphate metabolism is under active investigation. 30 Fungi and plants have sphingolipids and the major sphingosine contained in these organisms has the formula described below. It is known that these lipids WO 2008/089015 PCT/US2008/050695 4 have important roles in the cell growth of fungi and plants, but details of the roles remain to be solved. OH OH NH 2 5 | | |

H

3 C - (CH 2

)

12 - CH 2 - CH - CH - CH - CH 2 OH Recently it has been known that derivatives of sphingolipids and their related compounds exhibit a variety of biological activities through inhibition or 10 stimulation of the metabolism pathways. These compounds include inhibitors of protein kinase C, inducers of apoptosis, immuno-suppressive compounds, antifungal compounds, and the like. Substances having these biological activities are expected to be useful compounds for various diseases. 15 Derivatives of sphingosine have been prepared in various patents. For example, see U.S. Patents 4,952,683; 5,110,987; 6,235,912 B1 and 6,239,297 Bl. Also, compounds which are similar to certain spingosine derivatives, but which are not reported as being ligands for the spingosine receptors are reported in 20 various patents and published patent applications. See for example, U.S. Patents 5,294,722; 5,102,901; 5,403,851 and 5,580,878. U.S. Patent Application Publication No. U.S. 2003/0125371 A2. While certain of the compounds reported in the above patents are indoles, it does not appear that indole compounds have been reported as being ligands for sphingosine receptor 25 or having activity as sphingosine agonists or antagonists. SUMMARY OF THE INVENTION The present invention provides a derivative or analogue of sphingosine that is 30 able to regulate the functions of sphingolipid, and pharmaceutical compositions WO 2008/089015 PCT/US2008/050695 5 comprising said derivative or analogue. Compounds represented by the formula I having sphingosine- 1-phosphate receptor agonist and or antagonist biological activity: R1 4),/ _Ni IR3

Y(R

4 )n-A-(X)q zR2 A1-(XI)r'A2 B Formula I wherein: 10 R' R2, R3 and R 4 are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 carbons and 1 or 2 triple bonds, carbocyclic hydrocarbon groups having from 3 to 20 carbon atoms, heterocyclic groups having up to 20 carbon 15 atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, halo, CI to C 1 2 haloalkyl, hydroxyl, C 1 to C 1 2 alkoxy, C 3 to C 20 arylalkyloxy, C 1 to C 1 2 alkylcarbonyl, formyl, oxycarbonyl, carboxy, Ci to C 1 2 alkyl carboxylate, C 1 to C 1 2 alkyl amide, aminocarbonyl, amino, cyano, diazo, nitro, thio, sulfoxyl, and sulfonyl groups; 20 X and X' are independently selected from the group consisting of NR 5 , 0 and S;

R

5 is hydrogen, an alkyl group of 1 to 10 carbons, a cycloalkyl group of 5 to 10 carbons, phenyl or lower alkylphenyl; Y is a carbocyclic aryl or 25 heterocyclic aryl group wherein said carbocylic aryl comprises from 6 to 20 atoms and said heterocyclic aryl comprises from 2 to 20 carbon atoms and from 1 to 5 heteroatoms selected from the group consisting of WO 2008/089015 PCT/US2008/050695 6 nitrogen, oxygen and sulfur, and wherein said aryl may be bonded to A at any position; Z is O or S; n is 0 or an integer of from I to 5; o is 0 or an integer of from I to 3; p is 0 or an integer of from I to 3; q is 0 or 1; r is 0 or 1; A, A' and A 2 are independently selected from the group consisting of 10 (CH 2 )v wherein v is 0 or an integer of from 1 to 12, branched chain alkyl having 3 to 12 carbons, cycloalkyl having 3 to 12 carbons, alkenyl having 2 to 10 carbons and 1-3 double bonds and alkynyl having 2 to 10 carbons and 1 to 3 triple bonds; 67 B is selected from the group consisting of hydrogen, OR6, COOR , 921 136 7 15 NRR 9 , CONR R9, COR ", CH=NOR 11 , CH=NNR R' 3 wherein R6, R, R10 and R" are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 carbons and 1 or 2 triple bonds, a carbocyclic hydrocarbon group having 20 from 3 to 20 carbon atoms, a heterocyclic group having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, R 8 , R 9 , R1 2 and R1 3 are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 25 carbons and 1 or 2 triple bonds, a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, a heterocyclic group having up to 20 carbon WO 2008/089015 PCT/US2008/050695 7 atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, or R 8 and R 9 and/or R1 2 and R" , together, can form a divalent carbon radical of 2 to 5 carbons to form a heterocyclic ring with nitrogen, wherein any of 6 7 8 91 2 12 R6, R , R , R9, R', R", R or R' may be substituted with one or more 5 halogen, hydroxy, alkyloxy, cyano, nitro, mercapto or thiol radical; provided however, when v is 0, and r is 0, B is not hydrogen; or B is a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, or a heterocyclic group having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, and wherein when said B is a 10 carbocyclic or heterocyclic group B may be bonded to A2 at any position, or a pharmaceutically acceptable salt of said compound. 15 The aryl group is a carbocyclic aryl or heterocyclic aryl group wherein said 20 carbocylic aryl comprises from 6 to 20 atoms and said heterocyclic aryl comprise from 2 to 20 carbon atoms and from 1 to 5 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and preferably said aryl group is selected from the group consisting of benzene, pyridine, pyrazine, pyridazine, pyrimidine, triazine, thiophene, furan, thiazole, thiadiazole, 25 isothiazole, oxazole,oxadiazole, isooxazole, naphthalene, quinoline, tetralin, chroman, thiochroman, tetrahydroquinoline, dihydronaphthalene, tetrahydronaphthalen, chromene, thiochromene, dihydroquinoline, indan, dihydrobenzofuran, dihydrobenzothiophene, indene, benzofuran, benzothiophene, coumarin and coumarinone. Said aryl groups can be bonded to WO 2008/089015 PCT/US2008/050695 8 the above moiety at any position. Said aryl group may itself be substituted with any common organic functional group including but not limited to C 1 to C 1 2 alkyl, C 2 to C 6 alkenyl, C 2 to C 6 alkynyl, halo, C 1 to C 1 2 haloalkyl, hydroxyl, C 1 to C1 2 alkoxyl, CI to C 1 2 alkylcarbonyl, formyl, oxycarbonyl, carboxyl, C 1 to C 1 2 5 alkyl carboxylate, C, to C1 2 alkyl amide, aminocarbonyl, amino, cyano, diazo, nitro, thio, sulfoxyl, or sulfonyl groups. Preferably Z is 0. 10 Preferably, the carbocyclic aryl group will comprise from 6 to 14 carbon atoms, e.g. from 6 to 10 carbon atoms. Preferably the heterocyclic aryl group will comprise from 2 to 14 carbon atoms and one or more, e.g. from 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. 15 Preferably, A is CH 2 Preferably, X is NH. Preferably, n is 0 or an integer of 1 or 2 and R4 is fluoro. 20 Preferably, R' is i-propyl. Preferably, R 3 is selected from the group consisting of phenyl, which may be substituted with one or two flouro groups, and pyridyl. 25 Preferably, p is 0. Preferably, A' and A 2 are absent. Preferably, B is OR 6 or COOR 7 . 30 Preferably, X is 0, r is 1, A' is absent, A2 is (CH 2 )v, wherein v is 1 or 2, and B is

OR

6 or NRR 9 and R 6 , R 8 and R 9 are methyl.

WO 2008/089015 PCT/US2008/050695 9 Preferably, B is CR' 0

=NOR"R'

0 wherein R" 0 is H and R" is methyl or i-butyl or B is CONR 8

R

9 wherein R 8 and R9 are selected from the group consisting of H, methyl, ethyl and propyl, or R 8 and R 9 , together with N, form a 5-member ring. 5 Preferably, A' is absent, r is 0, A2 is CH 2 and B is OR 6 , wherein R 6 is H, or X is 0, r is 1 and B is COR' 0 ,wherein R1 0 is methyl. Specific Examples of the compounds of formula I include F N NF F / N N | rN F H N N /N N 010 o1 | NN -~~ ~ HN/'i~ N H N'9 N 1 N 1 0~ 11 N 0 10 N-N Some compounds within the scope of the invention may be prepared as depicted in Scheme 1. Thus, methyl 6-methoxyindole-2-carboxylate is treated with an electrophilic compound (e.g. benzyl bromide) in the presence of an weak base (e.g. potassium carbonate) to produce an N-alkylated indole (e.g. methyl 1 15 benzyl-6-methoxyindole-2-carboxylate). The 2-carboxylate group is converted to an alkyl group by a three-step process: Grignard reaction, elimination, and hydrogenation. The resulting 2-alkyl indole is carboxylated in the 3-position by treatment with dimethylformamide and phosphorus oxychloride followed by sodium hypochlorite oxidation of the resulting aldehyde. The carboxylic acid 20 may be further functionalized by treatment with an amine in the presence of N (3-dimethylaminopropyl)-N'-ethylcarbodimide (EDC) to produce a 6 methoxyindole-3-carboxamide derivative (e.g. 3,4-difluorophenylmethyl 6- WO 2008/089015 PCT/US2008/050695 10 methoxy-2-isopropyl-1-benzylindole-3-carboxamide). The carboxylic acid may also be treated with an alcohol or thiol in the presence of EDC to produce an ester and thiol ester derivatives, respectively. The 6-methoxy group may then be deprotected using boron tribromide and the resulting hydroxide subjected to 5 alkylating (e.g. cyclopentyl iodide/potassium carbonate) or acylating (e.g. pivaloyl chloride/ pyridine) reagents to produce a large variety of 6-substituted indole homologs and derivatives within the scope of the invention. Scheme 1

CO

2 Me 1) MeLi, THF

CO

2 Me BnBr, K 2

CO

3 MeO N 2) HCI-Et 2 O MeO N 3) H2, Pd-C H /h Ph 1) POCl 3 , DMF MeO

CO

2 H NH2 MeO N 2) NaCIO 2 , NaH 2

PO

4 ) t-BuOH,

H

2 0 N EDC, DMAP, Ph Ph/ CH 2

C

2 , 64% MeO \ \ N 1) BBr 3 , CH 2 Cl 2 N H F 2) c-C 5

H

9 1, K 2

CO

3 I H T F Ph F Ph F 10 Many other compounds within the scope of the invention may be prepared as depicted in Scheme 2. Thus, ethyl 4-iodobenzoate may be nitrated in the 3 position with fuming nitric acid and the resulting nitro compound reduced 15 under mild conditions (e.g. SnCl 2

-H

2 0) to produce ethyl 3-amino-4 iodobenzoate. This compound may be converted to the indole by treatment with a terminal alkyne (e.g. 3-methylbutyne) in the presence of a palladium catalyst and copper iodide followed by heating the aryl alkyne in the presence copper iodide. The resulting 2-alkyl indole may then be carbonylated in the 3-position 20 by treatment with dimethylformamide and phosphorus oxychloride and N alkylated as described above (benzyl bromide, potassium carbonate), followed by sodium hypochlorite oxidation to produce an N-alkylindole-3-carboxylic WO 2008/089015 PCT/US2008/050695 11 acid. The carboxylic acid may be further functionalized by treatment with an amine in the presence of EDC to Scheme 2 1HN H2SO4 I 1) (CH 3

)

2 CHCCH, Cul I ~.- 2) SnCIr-H 2 O ,a .. ~JII PdCI 2 (PPh 3

)

2 Et 3 N EtO 2 C EtO 2 C NH 2 2) Cul, DMF, 160 *C CO 2 H 1) DMF, POCI 3 1) (COCI) 2 , cat. DMF 2) BnBr, K 2 C0 3 EtO 2 C N 2) Amine, Et 3 N EtO 2 C H 3) NaCIO 2 , KH 2 PO4,. 3) LiOH, H 2 0-MeOH 0

RO

2 C / 0 N N CI) cat DMF KN N N N 1 ) Dibal-H, THF 00 2) TPAP-NMO N OHC / Amine, NaCNBH 3 N NaOAc, HOAc or N

H

2

N-OCH

3 / /0-N N $n N produce a 6-methoxyindole-3-carboxamide derivative (e.g. 3-pyridylmethyl 1 benzyl-6-carboethoxy-2-isopropylindole-3-carboxamide). The carboxylic acid 10 may also be treated with an alcohol or thiol in the presence of EDC to produce an ester and thiol ester derivatives, respectively. The 6-carboethoxy group may be further functionalized to produce a large variety of 6-substituted indole homologs and derivatives within the scope of the invention. For example, the 6 carboethoxy group be hydrolyzed with strong base and the resulting carboxylic 15 acid converted to the carboxylic acid chloride, which could be reacted with WO 2008/089015 PCT/US2008/050695 12 various alcohols or amines in the presence of base to produce ester or amide derivatives, respectively, such as 3-pyridylmethyll-benzyl-2-isopropyl-6-(1 pyrrolidinylcarbamoyl)indole-3-carboxamide. Alternatively, the 6-carboethoxy group could be reduced to an alcohol and re-oxidized to an aldehyde 5 intermediate, which may then be treated with an amine under reducing conditions to give amine derivatives such as 3-pyridylmethyl 1-benzyl-2 isopropyl-6-(1-pyrrolidinylmethyl)-indole-3-carboxamide. The aldehyde may also be treated with oxime or hydrazine compounds to produce oxime and hydrazone derivatives, respectively. Thus, many compounds within the scope of 10 the invention may be produced by the general route depicted in Scheme 2. DETAILED DESCRIPTION OF THE INVENTION Unless otherwise indicated, the following terms as used throughout this 15 specification have the following meanings: "Me" refers to methyl. "Et" refers to ethyl. 20 "tBu" refers to t-butyl. "iPr" refers to i-propyl. 25 "Ph" refers to phenyl. "Pharmaceutically acceptable salt" refers to those salts which retain the biological effectiveness and properties of the free bases and which are obtained by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, 30 sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.

WO 2008/089015 PCT/US2008/050695 13 "Alkyl" refers to a straight-chain, branched or cyclic saturated aliphatic hydrocarbon. Preferably, the alkyl group has 1 to 12 carbons. More preferably, it is a lower alkyl of from 1 to 7 carbons, most preferably 1 to 4 carbons. Typical alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary 5 butyl, pentyl, hexyl and the like. The alkyl group may be optionally substituted with one or more substituents are selected from the group consisting of hydroxyl, cyano, alkoxy, =0, =S, NO 2 , halogen, dimethyl amino and SH. "Alkenyl" refers to a straight-chain, branched or cyclic unsaturated hydrocarbon 10 group containing at least one carbon--carbon double bond. Preferably, the alkenyl group has 2 to 12 carbons. More preferably it is a lower alkenyl of from 2 to 7 carbons, most preferably 2 to 4 carbons. The alkenyl group may be optionally substituted with one or more substituents selected from the group consisting of hydroxyl, cyano, alkoxy, 0, S, NO 2 , halogen, dimethyl amino and 15 SH. "Alkynyl" refers to a straight-chain, branched or cyclic unsaturated hydrocarbon containing at least one carbon--carbon triple bond. Preferably, the alkynyl group has 2 to 12 carbons. More preferably it is a lower alkynyl of from 2 to 7 carbons, 20 most preferably 2 to 4 carbons. The alkynyl group may be optionally substituted with one or more substituents selected from the group consisting of hydroxyl, cyano, alkoxy, 0, S, NO 2 , halogen, dimethyl amino and SH. "Alkoxy" refers to an "O-alkyl" group. 25 "Aryl" refers to an aromatic group which has at least one ring having a conjugated pi electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups. The aryl group may be optionally substituted with one or more substituents selected from the group consisting of halogen, trihalomethyl, 30 hydroxyl, SH, OH, NO 2 , amine, thioether, cyano, alkoxy, alkyll, and amino.

WO 2008/089015 PCT/US2008/050695 14 "Alkaryl" refers to an alkyl that is covalently joined to an aryl group. Preferably, the alkyl is a lower alkyl. "Aryloxy" refers to an "O-aryl" group. 5 "Arylalkyloxy" refers to an "O-alkaryl" group. "Carbocyclic" refers to cyclic saturated or unsaturated aliphatic hydrocarbon and aryl hydrocarbon groups wherein the ring atoms are exclusively carbons, and comprises from 6 to 20 carbon atoms, including said ring atoms. 10 "Carbocyclic aryl" refers to an aryl group w erein the ring atoms are canbo "Heterocyclic" refers to cyclic groups wherein the ring atoms comprise carbon 15 atoms and at least one oxygen, nitrogen, and/or sulfur atom and may be saturated, unsaturated, i.e. have one or more double bonds, or aryl, and comprises up to 20 carbon atoms and from 1 to 5 of the above heteroatoms. "Heterocyclic aryl" refers to an aryl group having from 1 to 3 heteroatoms as 20 ring atoms, the remainder of the ring atoms being carbon. Heteroatoms include oxygen, sulfur, and nitrogen. "Hydrocarbyl" refers to a hydrocarbon radical having only carbon and hydrogen atoms. Preferably, the hydrocarbyl radical has from 1 to 20 carbon atoms, more 25 preferably from 1 to 12 carbon atoms and most preferably from 1 to 7 carbon atoms. "Substituted hydrocarbyl" refers to a hydrocarbyl radical wherein one or more, but not all, of the hydrogen and/or the carbon atoms are replaced by a halogen, 30 nitrogen, oxygen, sulfur or phosphorus atom or a radical including a halogen, nitrogen, oxygen, sulfur or phosphorus atom, e.g. fluoro, chloro, cyano, nitro, WO 2008/089015 PCT/US2008/050695 15 hydroxyl, phosphate, thiol, etc. "Amide" refers to --C(O)--NH--R', wherein R' is alkyl, aryl, alkylaryl or hydrogen. 5 "Ester" refers to --C(O)--O--R', wherein R' is alkyl, aryl or alkylaryl. "Thioamide" refers to --C(S)--NH--R', wherein R' is alkyl, aryl, alkylaryl or hydrogen. 10 "Thiol ester" refers to --C(O)--S--R', wherein R' is alkyl, aryl, alkylaryl or hydrogen. "Amine" refers to a --N(R")R"' group, wherein R" and R' are independently 15 selected from the group consisting of alkyl, aryl, and alkylaryl. "Thioether" refers to --S--R", wherein R" is alkyl, aryl, or alkylaryl. "Sulfonyl" refers to --S(0) 2 --R"", where R"" is aryl, C(CN)=C-aryl, CH 2 CN, 20 alkyaryl, sulfonamide, NH-alkyl, NH-alkylaryl, or NH-aryl. Also, alternatively the substituent on the phenyl moiety, as shown below, is referred to as an o, m or p substituent or a 2, 3 or 4 substituent, respectively. (Obviously, the 5 substituent is also a m substituent and the 6 substituent is an o 25 substituent.) Specific compounds of the invention, that are prepared according to Example 2 through 29 and/or Schemes 1 through 3, are able to inhibit the activity of sphingosine-1-phosphate receptors reported in Table I, below. Compounds were 30 assessed for their ability to activate or block activation of the human SIP3 receptor in T24 cells stably expressing the human 51 P3 receptor. Ten thousand WO 2008/089015 PCT/US2008/050695 16 cells/well were plated into 384-well poly-D-lysine coated plates one day prior to use. The growth media for the S1P3 receptor expressing cell line was McCoy's 5A medium supplemented with 10% charcoal-treated fetal bovine serum (FBS), 1% antibiotic-antimycotic and 400 pg/ml geneticin. On the day of the 5 experiment, the cells were washed twice with Hank's Balanced Salt Solution supplemented with 20 mM HEPES (HBSS/Hepes buffer). The cells were then dye loaded with 2 uM Fluo-4 diluted in the HBSS/Hepes buffer with 1.25 mM Probenecid and incubated at 37 0 C for 40 minutes. Extracellular dye was removed by washing the cell plates four times prior to placing the plates in the 10 FLIPR (Fluorometric Imaging Plate Reader, Molecular Devices). Ligands were diluted in HBSS/Hepes buffer and prepared in 384-well microplates. The positive control, Sphingosine-1-Phosphate (SIP), was diluted in HBSS/Hepes buffer with 4 mg/ml fatty acid free bovine serum albumin. The FLIPR transferred 12.5 pl from the ligand microplate to the cell plate and took 15 fluorescent measurements for 75 seconds, taking readings every second, and then for 2.5 minutes, taking readings every 10 seconds. Drugs were tested over the concentration range of 0.61 nM to 10,000 nM. Data for Ca+ 2 responses were obtained in arbitrary fluorescence units and not translated into Ca+ 2 concentrations. IC 50 values were determined through a linear regression 20 analysis using the Levenburg Marquardt algorithm. Table 1 S1P3 Compound Structure inh) Number (% inh) Kb F F N\/ 560 nM 7 ( 98) F 8 N; ,N' 3.1 M 0 (71) WO 2008/089015 PCT/US2008/050695 17 F F 9 \/; iN 1 ~ ln (100) F N 2 nM 10 HN/~/ (100) I2 n F - - /_6 n M 11 H N 1 (100) 2 nM F 3 nM 12 H (100) ND F F 1.H nM 13 N/(100) 0 I,. 1.6 nM Table I S1P3 Compound Stnicture * Number (% inh) Kb F F #H ~ 3.1 gM (96) 2 nM 16F 15 d ii(100) O H~ 3nM 16 N (100) o1.2 nM WO 2008/089015 PCT/US2008/050695 18 F F 17 6.5 nM 17 (99) 18 , 242 nM o II I(94) F 19~~~ -NINnM 19 (100) 20!f~ (100) 0a Table 1, SI P3 Compound Structure (%inh) Number (%Kbh 21 N ,-N \/ 2n 00 F) F 22 H 4~> 4nM 22 N! ~ (99) ".0 23 14 N / InM 6 ~ (100) WO 2008/089015 PCT/US2008/050695 19 F 24 20OnM 6 'K ~ (100) 25 N/ 14nM o 1 (100) 39 nM 26 (100) 6o13 nM F 6 nM 27 (100) 11 nM The compounds of Table 1B are prepared according to procedures 5 analogous to the procedures of Schemes 1 through 3 and/or Examples 2 through 29. These compounds are also tested for ability to inhibit the activity of the S1 P3 receptor. Table 1B 10 S1P3 Compound Structure inh) Number (% inh) Kb F F HN / NnM (100) 0~ 8 nM 0 FO F :H ,N' 12 nM 0Z) (100) WO 2008/089015 PCT/US2008/050695 20 F F N 0 OH (40) . F H;N N N 170 nM 0 (100) 0 F H F -N 197 nM 0 (100) r 0 o N (100) Table 1 B S1P3 Compound Structure Inh0 Number (% inh) Kb F F F /N 346 nM N ~- N~Z (99) F NF$$R ~ N H(99) WO 2008/089015 PCT/US2008/050695 21 F F "' N / ~362 nM (93) FN 0 o (98) F N HN FZ\ N 2.3 pM o _______ H(84) 0N F F 2.6 pLM O H~ N p I H(8 7 ) Table 18B S1 P3 Compound Structure IC5( Number (% inh) Kb F N (97) F Number N 0(0nM o 0 (97) F9 F 0100 WO 2008/089015 PCT/US2008/050695 22 H 0.9 nM ~ K (100) F Ft H 21 M o I (99) N 88 nM 6 > (100) ~ q 00(95) Table lB Compound SIP3 ICs 0 Number Structure (% inh) Kb F NH NN 9 nM o (100) H N N ,N 9 9nM (96) 0 N, 187 nM o6 0-1 (95) 0 FF N; 1 \/ 45 nM 0 (99) WO 2008/089015 PCT/US2008/050695 23 NN H/ ND (98) H, 1.2 p o I(99) 0 N 4EN o (99) N N ND o H (54)

OHD

WO 2008/089015 PCT/US2008/050695 24 Table 1B S1P3 Compound Structure IC5h Number (% inh) Kb F \/H /42 nM (99) F N 43 nM 0 145 nM o K' (99) 0 H NO 145 nM o ~ ~N~J< (99) (10 F "" /~ /262 nM 0 (98) F\ H 147 nM NN Tae (100) F, H / __ 33 nM Q N h(100) NN 7 nM 5 Table 1B WO 2008/089015 PCT/US2008/050695 25 S1P3 Compound Structure Inh0 Number (% inh) Kb F F 673nRM N P (100) F F N/N -H j 2.4 M N;,/ (80) 0 H (10 174 nM (100) F F\ I 58nM N N N o 0 (100) R F 19 nM H; (100) F0 3 nM / ~ 44nM 0 Table 1B 5 Compound Structure 0 Number S(% inh) WO 2008/089015 PCT/US2008/050695 26 Kb F H/N 288 nM N N (100) o o F N- N 0, H / N324 nM (99) H~ 5nM N O/\/ (100) 9 nM F N o I ~ (100) F 15 nM 13 nM H N NA (65) FN \I ~/N93 nM (100) N/ Table 1B S1P3 Compound Structure ICSo Number (% inh) Kb WO 2008/089015 PCT/US2008/050695 27 F H\C 137 nM (100) As a result of the above activity of the compounds utilized in the method of the present invention, it is clear that such compounds may be used in treating the following diseases and conditions for the following reasons. 5 Glaucoma SIP3 subtypes are expressed in primary human trabecular meshwork cells and SIP decreases outflow facility >30% in perfused porcine eyes (See IOVS 45, 2263; 2004) by altering paracellular permeability. 10 Dry Eye/Immunology Induces lymphocyte sequestration without affecting T cell proliferation. Angiogenesis disorders S IP3 receptor subtype is expressed in vascular endothelial cells and 15 siRNA knockdown of S1 P1 and Si P3 inhibits angiogenesis. SIP also promotes vascular endothelial cell migration and promotes barrier assembly and integrity. Cardiovascular (S1P3) Si P3 "knock out" mice lack Si P induced pulmonary edema. 20 The invention is further illustrated by the following examples which are illustrative of a specific mode of practicing the invention and are not intended as limiting the scope of the claims. Unless otherwise indicated, the following Chemical Abbreviations are used in 25 the examples: BBr 3 : boron tribromide HCl: hydrogen chloride or hydrochloric acid KOH: potassium hydroxide WO 2008/089015 PCT/US2008/050695 28

K

2 C0 3 : potassium carbonate

KH

2

PO

4 : potassium dihydrogenphosphate NaOH: sodium hydroxide NaHC03: sodium bicarbonate 5 NaClO 2 : sodium hypochlorite Nal: sodium iodide Na 2

SO

4 : sodium sulfate MgSO 4 : magnesium sulfate POCl 3 : phosphorus oxychloride 10 t-BuOH: tert-butyl alcohol MeOH: methanol EtOH: ethanol i-PrOH: isopropanol EtOAc: ethyl acetate 15 Et 2 0: diethyl ether

CH

2 Cl 2 : methylene chloride

CH

3 CN: acetonitrile DHP: dihydropyran DMAP: 4-(dimethylamino)pyridine 20 DMF: NN-dimethylformamide DMSO: dimethylsulfoxide EDC: 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide LDA: lithium diisopropylamide LiAlH4: lithium aluminum hydride 25 MOMCl: methyl chloromethyl ether MeLi: methyllithium MeMgBr: Methylmagnesium bromide NaBH 3 CN: sodium cyanoborohydride NMO: 4-methylmorpholine, N-oxide 30 Pd-C: palladium on activated carbon PhCHO: benzaldehyde WO 2008/089015 PCT/US2008/050695 29 THF: tetrahydrofuran THP: tetrahydropyran TPAP: tetrapropylammonium perruthenate PTLC: preparative thin layer chromatography 5 Acetyl chloride, benzyl bromide, 2-bromoethyl methyl ether, cyclopentyl iodide, diisopropylethylamine, 2-dimethylaminoethyl chloride hydrochloride, dimethylcarbamyl chloride, 1-iodobutane, 2-iodobutane, iodoethane, 1 iodohexane, 1-iodopropane, 2-iodopropane, 4-methylbenzene-1sulfonyl chloride, pivaloyl chloride, pyridinium p-toluenesufonate and tetrahydrofuran-3 10 ol were purchased from Aldrich Chemical Company. Scheme 3 ' I C 2 Me -

C

2 Me N~-~ MeO N MeO MeO OH Ph Ph 1 2 3 / CHO C0 2 H MeO>iv MeO v MeO H Ph) Ph Ph 4 5 6 MeO /\ O HO \ 0 - N N F xorxi P2 F Ph-a 7 8 Viii RO \xi RO / I H 00 HI PhF R O P-- N F PhF 0/ Ph' F 9 R= Et 16 R = cydopentyl N 10 = -P 17R =CHCH 2 CN H I26 R =methoxymethyl 0R =-Pr CH2OCH3 F 27 R = 2-tetrahydrofuranyl 11 R = i-Pr 18 R =cCHNcH) P-j' 12 R = n-Bu F 13 R = i-Bu 19 R0= 14 R = hexyl 21 R = N(CH 3

)

2 15 R = Bn 20 R = 22 R = t-Bu 23 R=Me 0 24 R = Et 25 R = i-Pr WO 2008/089015 PCT/US2008/050695 30 a Reagents and conditions: (i) BnBr, K 2 C0 3 , DMF; (ii) MeLi, THF; (iii) H 2 , Pd C, EtOAc, EtOH, HCl-Et 2 O; (iv) POCl 3 , DMF; (v) NaClO 2 , KH 2

PO

4 , isobutene, t-BuOH, CH 3 CN, H20; (vi) 3,4-difluorobenzylamine, EDC, DMAP,

CH

2 Cl 2 ; (vii) BBr 3 , CH 2 Cl 2 ; (viii) RX, K 2 C0 3 , DMF; (ix) RCOCl, pyridine; (x) 5 MOMCl, i-Pr 2 NEt, CH 2 Cl 2 ; (xi) 2,3-dihydrofuran, PPTS, CH 2 Cl 2 . Example 2 Methyl 1-Benzyl-6-methoxy-1H-indole-2-carboxylate (Compound 2). To a solution of methyl 6-methoxy-1H-indole-2-carboxylate (Compound 1, 1.0 g, 10 4.9 mmol) in DMF (10 ml) was added K 2 C0 3 (2.0 g, 14.6 mmol) and benzyl bromide (0.87 ml, 7.3 mmol). The mixture was stirred at room temperature for 40 h and was diluted with EtOAc, washed with H20, brine, dried over Na 2

SO

4 and concentrated in vacuo. The residue was purified by crystallization from Et 2 0 to yield the title compound as an off-white solid. 15 'H NMR (500 MHz, CHLOROFORM-d) 6 ppm 3.81 (s, 3 H), 3.85 (s, 3 H), 5.81 (s, 2 H), 6.73 (d, J = 2.0 Hz, 1 H), 6.84 (dd, J = 8.8, 2.0 Hz, 1 H), 7.07 (d, J= 6.8 Hz, 2 H), 7.19 - 7.29 (m, 3 H), 7.33 (s, 1 H), 7.58 (d, J= 8.8 Hz, 1 H). Example 3 20 2-(1-Benzyl-6-methoxy-1H-indol-2-yl)propan-2-ol (Compound 3). To a solution of methyl 1-benzyl-6-methoxy- 1 H-indole-2-carboxylate (Compound 2, 4.33 g, 14.7 mmol) in THF (50 ml) at 0 *C under argon was added MeLi (3.0 M in diethoxymethane, 19.6 ml, 58.7 mmol) slowly. After 1 h, the ice-water bath was removed and the reaction was stirred at room temperature for lh, cooled to 25 -78 *C, quenched with dry ice, diluted with EtOAc, washed with H20, brine, dried over Na 2

SO

4 , concentrated in vacuo to yield the crude title compound as a yellow solid. 'H NMR (500 MHz, CHLOROFORM-d) 6 ppm 1.69 (s, 6 H), 3.73 (s, 3 H), 5.76 (s, 2 H), 6.42 (s, 1 H), 6.55 (d, J= 2.4 Hz, 1 H), 6.75 - 6.81 (m, 1 H), 6.96 30 (d, J = 7.3 Hz, 2 H), 7.22 (d, J = 7.3 Hz, 1 H), 7.25 - 7.30 (m, 2 H), 7.49 (d, J= 8.8 Hz, 1 H). Example 4 WO 2008/089015 PCT/US2008/050695 31 1-Benzyl-2-isopropyl-6-methoxy-1H-indole (Compound 4). To a solution of 2-(1-benzyl-6-methoxy-1H-indol-2-yl)propan-2-ol (Compound 3, 1.05 g, 3.57 mmol) in EtOAc (35 ml) and EtOH (15 ml) was added 10% Pd-C (190 mg, 0.18 mmol) and HCl-Et 2 O (1.0 M, 1.25 ml, 1.25 mmol). The mixture was stirred 5 under hydrogen gas (atmospheric pressure) for lh and was filtered. To the filtrate was added NaHCO 3 (0.5 g) and H 2 0 (0.5 ml), followed by Na 2

SO

4 and MgSO 4 . This was then filtered and concentrated in vacuo to yield the crude title compound as a yellow solid. 'H NMR (300 MHz, CHLOROFORM-d) 8 ppm 1.31 (d, J = 6.7 Hz, 6 H), 2.90 10 - 3.10 (in, 1 H), 3.79 (s, 3 H), 5.33 (s, 2 H), 6.33 (s, 1 H), 6.68 (d, J = 2.1 Hz, 1 H), 6.79 (dd, J = 8.5, 2.3 Hz, 1 H), 6.94 - 7.04 (in, 2 H), 7.20 - 7.37 (in, 2 H), 7.49 (d, J = 8.5 Hz, 1 H). Example 5 15 1-Benzyl-2-isopropyl-6-methoxy-1H-indole-3-carbaldehyde (Compound 5). POC1 3 (0.48 ml, 5.23 mmol) was added dropwise to anhydrous DMF (2 ml) at 0 'C under argon. After stirred for 30 min, this solution was added dropwise to a solution of 1-benzyl-2-isopropyl-6-methoxy-1H-indole (Compound 4, 583 mg, 2.09 mmol) in anhydrous DMF (8 ml) at 0 *C under argon. The reaction was 20 stirred for 1 h at 0 'C and 30 min at room temperature, diluted with EtOAc, washed with aqueous NaHCO 3 , brine, dried over Na 2

SO

4 , and concentrated in vacuo. The residue was purified by chromatography on silica gel (0-+30% EtOAc-hexanes) to yield the title compound as a light yellow syrup. 'H NMR (500 MHz, CHLOROFORM-d) 8 ppm 1.45 (d, J= 7.3 Hz, 6 H), 3.40 25 - 3.52 (in, 1 H), 3.79 (s, 3 H), 5.40 (s, 2 H), 6.69 (d, J = 2.4 Hz, 1 H), 6.94 (dd, J = 8.8, 2.0 Hz, 1 H), 7.01 (d, J = 7.3 Hz, 2 H), 7.25 - 7.35 (in, 3 H), 8.28 (d, J 8.8 Hz, 1 H), 10.45 (s, 1 H). Example 6 30 1-Benzyl-2-isopropyl-6-methoxy-1H-indole-3-carboxylic Acid (Compound 6). To a solution of 1-benzyl-2-isopropyl-6-methoxy-IH-indole-3-carbaldehyde WO 2008/089015 PCT/US2008/050695 32 (Compound 5, 608 mg, 1.98 mmol) in t-BuOH (15 ml), CH 3 CN (15 ml), and 2 methyl-2-butene (10 ml) was added a solution of KH 2

PO

4 (5.4 g, 39.6 mmol) and NaClO 2 (80%, 4.5 g, 39.6 mmol) in H 2 0 (50 ml). The mixture was stirred at room temperature and additional 2-methyl-2-butene, KH 2

PO

4 , and NaClO 2 were 5 added at the above ratio every 16-24 h until the starting material was consumed. The reaction mixture was extracted with EtOAc (x3) and the combined organic layer was washed with brine, dried over Na 2

SO

4 , and concentrated in vacuo. The residue was purified by chromatography on silica gel (0-+25% EtOAc hexanes) to yield the title compound as a yellow solid. 10 'H NMR (500 MHz, CHLOROFORM-d) 8 ppm 1.39 (d, J = 7.3 Hz, 6 H), 3.75 (s, 3 H), 3.99 - 4.17 (m, 1 H), 5.45 (s, 2 H), 6.62 (d, J = 2.4 Hz, 1 H), 6.90 (dd, J = 8.8, 2.4 Hz, 1 H), 6.99 (d, J = 7.3 Hz, 2 H), 7.22 - 7.34 (m, 3 H), 8.18 (d, J 8.8 Hz, 1 H). 15 Example 7 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-methoxy-1H-indole-3 carboxamide (Compound 7). To a solution of 1-benzyl-2-isopropyl-6 methoxy-1H-indole-3-carboxylic acid (Compound 6, 226 mg, 0.70 mmol) in

CH

2 Cl 2 (7.0 ml) was added EDC (202 mg, 1.05 mmol) and DMAP (128 mg, 20 1.05 mmol) followed by 3,4-difluorobenzylamine (0.25 ml, 2.1 mmol). The reaction was stirred at room temperature for 18 h, diluted with EtOAc, washed with H 2 0, brine, dried over Na 2

SO

4 and concentrated in vacuo. The residue was purified by chromatography on silica gel (0--*30% EtOAc-hexanes) to yield the title compound as a yellow solid. 25 'H NMR (500 MHz, CHLOROFORM-d) 8 ppm 1.37 (d, J = 7.3 Hz, 6 H), 3.65 - 3.73 (m, 1 H), 3.74 (s, 3 H), 4.66 (d, J = 5.9 Hz, 2 H), 5.40 (s, 2 H), 6.30 (t, J = 6.3 Hz, 1 H), 6.63 (d, J = 2.0 Hz, 1 H), 6.82 (dd, J = 8.8, 2.4 Hz, 1 H), 6.96 (d, J= 6.8 Hz, 2 H), 7.11 - 7.17 (m, 2 H), 7.21 - 7.31 (m, 4 H), 7.51 (d, J:- 8.3 Hz, 1 H). 30 Example 8 WO 2008/089015 PCT/US2008/050695 33 1-Benzyl-N-(3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3 carboxamide (Compound 8). To a solution of 1-benzyl-N-(3,4 difluorobenzyl)-2-isopropyl-6-methoxy-1H-indole-3-carboxamide (Compound 7, 452 mg, 1.0 mmol) in CH 2 C1 2 (20 ml) at 0 *C was added BBr 3 (1.0 M in 5 CH 2 Cl 2 , 3.0 ml, 3.0 mmol) dropwise. The reaction was stirred for 1 h at 0 'C and 1 h at room temperature, quenched with ice, extracted with EtOAc, the organic layer was washed with brine, dried over Na 2

SO

4 , and concentrated in vacuo. The residue was purified by chromatography on silica gel (0-+50% EtOAc-hexanes) to yield the title compound as a yellow solid. 10 'H NMR (500 MHz, CHLOROFORM-d) 8 ppm 1.37 (d, J= 7.3 Hz, 6 H), 3.65 - 3.74 (in, 1 H), 4.66 (d, J = 5.9 Hz, 2 H), 4.78 (s, 1 H), 5.37 (s, 2 H), 6.27 (t, J = 5.6 Hz, 1 H), 6.60 (d, J = 2.4 Hz, 1 H), 6.71 (dd, J = 8.5, 2.2 Hz, 1 H), 6.95 (d, J = 6.8 Hz, 2 H), 7.11 - 7.17 (in, 2 H), 7.21 - 7.32 (in, 4 H), 7.46 (d, J = 8.8 Hz, 1 H). 15 Example 9 1-benzyl-N-(3,4-difluorobenzyl)-6-ethoxy-2-isopropyl-1H-indole-3 carboxamide (Compound 9). General Procedure A. To a solution of 1 benzyl-N-(3,4-difluorobenzyl)-6-hydroxy-2-isopropyl- 1 H-indole-3 -carboxamide 20 (Compound 8, 40 mg, 0.092 mmol) in DMF (2.0 ml) was added K 2 C0 3 (39 mg, 0.28 mmol) and iodoethane (22 pd, 0.28 mmol). The reaction was stirred at room temperature for 48 h, diluted with EtOAc, washed with H 2 0, brine, dried over Na 2

SO

4 and concentrated in vacuo. The residue was purified by PTLC on silica gel (30% EtOAc-hexanes) to yield the title compound as an off-white 25 solid. 'H NMR (500 MHz, CHLOROFORM-d) 6 ppm 1.37 (t, J= 7.0 Hz, 3 H), 1.38 (d, J = 7.3 Hz, 6 H), 3.68 - 3.75 (in, 1 H), 3.96 (q, J = 7.0 Hz, 2 H), 4.67 (d, J= 6.3 Hz, 2 H), 5.40 (s, 2 H), 6.31 (t, J 5.4 Hz, 1 H), 6.64 (d, J = 2.4 Hz, 1 H), 6.82 (dd, J= 8.8, 2.0 Hz, 1 H), 6.97 (d, J= 6.8 Hz, 2 H), 7.13 - 7.17 (in, 2 H), 30 7.23 - 7.31 (m, 4 H), 7.52 (d, J= 8.3 Hz, 1 H) WO 2008/089015 PCT/US2008/050695 34 Example 10 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-propoxy-1H-indole-3 carboxamide (Compound 10). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide 5 (Compound 8, 8.0 mg, 0.018 mmol) in DMF (1.0 ml) was reacted with K 2 C0 3 (8.0 mg, 0.055 mmol) and 1-iodopropane (9.0 pl, 0.092 mmol) to yield the title compound as a white solid. H NMR (500 MHz, METHANOL-d 4 ) 8 ppm 0.99 (t, J = 7.6 Hz, 3 H), 1.32 (d, J = 7.3 Hz, 6 H), 1.67 - 1.77 (m, 2 H), 3.42 - 3.53 (m, 1 H), 3.84 (t, J = 6.6 Hz, 10 2 H), 4.57 (s, 2 H), 5.46 (s, 2 H), 6.73 (d, J = 2.0 Hz, 1 H), 6.78 (dd, J = 8.8, 2.4 Hz, 1 H), 6.95 (d, J = 6.8 Hz, 2 H), 7.19 - 7.29 (m, 5 H), 7.30 - 7.36 (m, 1 H), 7.49 (d, J = 8.3 Hz, 1 H). Example 11 15 1-Benzyl-N-(3,4-difluorobenzyl)-6-isopropoxy-2-isopropyl-1H-indole-3 carboxamide (Compound 11). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 8.0 mg, 0.018 mmol) in DMF (1.0 ml) was reacted with K 2 CO3 (8.0 mg, 0.055 mmol) and 2-iodopropane (9.0 pl, 0.092 mmol) to yield the title 20 compound as a white solid. 'H NMR (500 MHz, METHANOL-d 4 ) 8 ppm 1.21 (d, J = 5.9 Hz, 6 H), 1.33 (d, J = 7.3 Hz, 6 H), 3.45 - 3.55 (m, 1 H), 4.41 - 4.50 (m, 1 H), 4.57 (s, 2 H), 5.46 (s, 2 H), 6.72 (d, J = 2.0 Hz, 1 H), 6.74 - 6.79 (m, 1 H), 6.96 (d, J = 7.3 Hz, 2 H), 7.18 - 7.29 (m, 5 H), 7.30 - 7.37 (m, 1 H), 7.49 (d, J= 8.8 Hz, 1 H). 25 Example 12 1-Benzyl-6-butoxy-N-(3,4-difluorobenzyl)-2-isopropyl-1H-indole-3 carboxamide (Compound 12). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide 30 (Compound 8, 10.7 mg, 0.025 mmol) in DMF (1.0 ml) was reacted with K 2 CO3 WO 2008/089015 PCT/US2008/050695 35 (10.0 mg, 0.074 mmol) and 1-iodobutane (14.0 pl, 0.12 mmol) to yield the title compound as a white solid. 'H NMR (500 MHz, CHLOROFORM-d) 8 ppm 0.93 (t, J = 7.3 Hz, 3 H), 1.37 (d, J= 7.3 Hz, 6 H), 1.40 - 1.50 (m, 2 H), 1.66 - 1.74 (m, 2 H), 3.61 - 3.75 (m, 1 5 H), 3.88 (t, J: 6.6 Hz, 2 H), 4.66 (d, J 6.3 Hz, 2 H), 5.39 (s, 2 H), 6.30 (t, J 5.9 Hz, 1 H), 6.63 (d, J = 2.0 Hz, 1 H), 6.81 (dd, J = 8.5, 2.2 Hz, 1 H), 6.96 (d, J= 6.8 Hz, 2 H), 7.10 - 7.17 (m, 2 H), 7.21 - 7.32 (m, 4 H), 7.50 (d, J= 8.8 Hz, 1 H). 10 Example 13 1-Benzyl-N-(3,4-difluorobenzyl)-6-isobutoxy-2-isopropyl-1H-indole-3 carboxamide (Compound 13). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 10.7 mg, 0.025 mmol) in DMF (1.0 ml) was reacted with K 2 C0 3 15 (10.0 mg, 0.074 mmol) and 2-iodobutane (14.0 pl, 0.12 mmol) to yield the title compound as a white solid. 'H NMR (500 MHz, CHLOROFORM-d) 8 ppm 0.98 (d, J = 6.8 Hz, 6 H), 1.36 (d, J= 7.3 Hz, 6 H), 1.96 - 2.08 (m, 1 H), 3.65 (d, J= 6.8 Hz, 2 H), 3.65 - 3.72 (m, 1 H), 4.66 (d, J = 6.3 Hz, 2 H), 5.39 (s, 2 H), 6.29 (t, J = 5.6 Hz, 1 H), 6.63 20 (d, J = 2.0 Hz, 1 H), 6.82 (dd, J = 8.8, 2.0 Hz, 1 H), 6.96 (d, J = 6.8 Hz, 2 H), 7.11 - 7.16 (m, 2 H), 7.21 - 7.31 (m, 4 H), 7.50 (d, J= 8.8 Hz, 1 H). Example 14 1-Benzyl-N-(3,4-difluorobenzyl)-6-(hexoxy)-2-isopropyl-1H-indole-3 25 carboxamide (Compound 14). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 10.7 mg, 0.025 mmol) in DMF (1.0 ml) was reacted with K 2 C0 3 (10.0 mg, 0.074 mmol) and 1-iodohexane (18.0 pL, 0.12 mmol) to yield the title compound as a white solid. 30 'H NMR (500 MHz, CHLOROFORM-d) S ppm 0.85 - 0.93 (m, 3 H), 1.24 1.33 (m, 4 H), 1.37 (d, J= 6.8 Hz, 6 H), 1.38 - 1.46 (m, 2 H), 1.66 - 1.77 (m, 2 WO 2008/089015 PCT/US2008/050695 36 H), 3.63 - 3.75 (m, 1 H), 3.87 (t, J = 6.6 Hz, 2 H), 4.66 (d, J = 5.9 Hz, 2 H), 5.39 (s, 2 H), 6.30 (t, J = 5.6 Hz, 1 H), 6.63 (d, J = 2.4 Hz, 1 H), 6.81 (dd, J 8.8, 2.4 Hz, 1 H), 6.96 (d, J= 6.8 Hz, 2 H), 7.10 - 7.16 (m, 2 H), 7.21 - 7.31 (m, 4 H), 7.50 (d, J = 8.8 Hz, 1 H). 5 Example 15 1-Benzyl-6-(benzyloxy)-N-(3,4-difluorobenzyl)-2-isopropyl-1H-indole-3 carboxamide (Compound 15). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl- 1 H-indole-3-carboxamide 10 (Compound 8, 10.7 mg, 0.025 mmol) in DMF (1.0 ml) and acetone (1.0 ml) was reacted with K 2

CO

3 (10.0 mg, 0.074 mmol), benzyl bromide (14.0 pl, 0.12 mmol), and catalytic amount of NaI to yield the title compound as an off-white solid. 'H NMR (500 MHz, CHLOROFORM-d) 6 ppm 1.37 (d, J = 7.3 Hz, 6 H), 3.65 15 - 3.75 (m, 1 H), 4.66 (d, J = 6.3 Hz, 2 H), 4.99 (s, 2 H), 5.37 (s, 2 H), 6.28 (t, J = 6.3 Hz, 1 H), 6.71 (d, J = 2.0 Hz, 1 H), 6.89 (dd, J = 8.8, 2.0 Hz, 1 H), 6.95 (d, J= 6.8 Hz, 2 H), 7.11 - 7.18 (m, 2 H), 7.22 - 7.30 (m, 5 H), 7.31 - 7.39 (m, 4 H), 7.51 (d, J = 8.8 Hz, 1 H). 20 Example 16 1-Benzyl-6-(cyclopentoxy)-N-(3,4-difluorobenzyl)-2-isopropyl-1H-indole-3 carboxamide (Compound 16). Following General Procedure A, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 40 mg, 0.092 mmol) in DMF (1.0 ml) was reacted with K 2 CO3 25 (38 mg, 0.28 mmol), cyclopentyl iodide (53 p, 0.46 mmol) to yield the title compound as a white solid. 'H NMR (300 MHz, CHLOROFORM-d) 8 ppm 1.37 (d, J = 7.0 Hz, 6 H), 1.48 - 1.60 (m, 2 H), 1.66 - 1.86 (m, 6 H), 3.62 - 3.83 (m, 1 H), 4.56 - 4.77 (m, 3 H), 5.38 (s, 2 H), 6.32 (t, J= 5.9 Hz, 1 H), 6.61 (d, J= 2.1 Hz, 1 H), 6.78 (dd, J= 30 8.8, 2.1 Hz, 1 H), 6.91 - 7.02 (m, 2 H), 7.08 - 7.17 (m, 2 H), 7.17 - 7.36 (m, 4 H), 7.49 (d, J = 8.5 Hz, 1 H).

WO 2008/089015 PCT/US2008/050695 37 Example 17 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-(2-methoxyethoxy)-1H indole-3-carboxamide (Compound 17). Following General Procedure A, 1 5 benzyl-N-(3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 17 mg, 0.039 mmol) in DMF (1.0 ml) was reacted with K 2 C0 3 (28 mg, 0.20 mmol), 2-bromoethyl methyl ether (18 pl, 0.20 mmol) to yield the title compound (9 mg, 49%). 'H NMR (300 MHz, CDCl 3 ) 8 ppm 1.37 (d, J = 7.04 Hz, 6 H), 3.40 (s, 3 H), 10 3.60 - 3.78 (m, 3 H), 4.04 (dd, J = 5.42, 3.96 Hz, 2 H), 4.66 (d, J = 5.86 Hz, 2 H), 5.39 (s, 2 H), 6.30 (t, J = 5.86 Hz, 1 H), 6.68 (d, J = 2.35 Hz, 1 H), 6.85 (dd, J= 8.65, 2.20 Hz, 1 H), 6.89 - 7.01 (m, 2 H), 7.10 - 7.18 (m, 2 H), 7.17 7.35 (m, 4 H), 7.51 (d, J = 8.79 Hz, 1 H). 15 Example 18 1-Benzyl-N-(3,4-difluorobenzyl)-6-(2-(dimethylamino)ethoxy)-2-isopropyl 1H-indole-3-carboxamide (Compound 18). Following General Procedure A, 1-benzyl-N-(3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3 carboxamide (Compound 8, 17 mg, 0.039 mmol) in DMF (1.0 ml) was reacted 20 with K 2

CO

3 (28 mg, 0.20 mmol), 2-dimehtylamino ethyl chloride hydrochloride (20 mg, 0.20 mmol) to yield the title compound (10 mg, 53%). 'H NMR (300 MHz, CD 3 0D) 8 ppm 1.32 (d, J = 7.04 Hz, 6 H), 2.30 (s, 6 H), 2.71 (t, J = 5.42 Hz, 2 H), 3.37 - 3.59 (m, 1 H), 4.02 (t, J = 5.42 Hz, 2 H), 4.57 (s, 2 H), 5.48 (s, 2 H), 6.73 - 6.88 (m, 2 H), 6.89 - 7.02 (m, 2 H), 7.12 - 7.40 (m, 25 6 H), 7.50 (d, J = 8.50 Hz, 1 H). Example 19 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-(tetrahydrofuran-3-yloxy) 1H-indole-3-carboxamide (Compound 19). To a solution of 1-benzyl-N-(3,4 30 difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 8 mg, 0.039 mmol) in DMF (1.0 ml) was added K 2

CO

3 (13 mg, 0.092 mmol) WO 2008/089015 PCT/US2008/050695 38 and catalytic amount of NaOH, 3-iodotetrahydrofuran (Compound 29, 120 mg, crude). The reaction was stirred at room temperature for 2 days, and purified by a short silica gel column to yield the title compound (8 mg, 86%). 'H NMR (300 MHz, CDCl 3 ) 8 ppm 1.38 (d, J = 7.04 Hz, 6 H), 1.95 - 2.14 (m, 2 5 H), 3.59 - 4.01 (m, 5 H), 4.66 (d, J- 6.16 Hz, 2 H), 4.74 - 4.88 (m, 1 H), 5.39 (s, 2 H), 6.29 (t, J = 4.40 Hz, 1 H), 6.57 (d, J = 2.05 Hz, 1 H), 6.69 - 6.83 (m, 1 H), 6.96 (d, J= 7.62 Hz, 2 H), 7.08 - 7.19 (m, 2 H), 7.18 - 7.35 (m, 4 H), 7.51 (d, J = 8.79 Hz, 1 H). 10 Example 20 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-(2-oxotetrahydrofuran-3 yloxy)-1H-indole-3-carboxamide (Compound 20). Following General Procedure A, 1 -benzyl-N-(3,4-difluorobenzyl)-6-hydroxy-2-isopropyl- 1 H indole-3-carboxamide (Compound 8, 19mg, 0.044mol) in DMF (1.0 ml) was 15 reacted with K 2 C0 3 (30 g, 0.22 mmol), 3-bromodihydrofuran-2(3H)-one (20 mg, 0.22mmol) to yield the title compound (16mg, 71%). 1H NMR (300 MHz, acetone-d 6 ) 8 ppm 1.33 (d, J = 5.57 Hz, 6 H), 2.21 - 2.42 (m, 1 H), 2.68 - 2.88 (m, 1 H), 3.43 - 3.65 (m, 1 H), 4.21 - 4.53 (m, 2 H), 4.66 (d, J = 6.16 Hz, 2 H), 5.10 - 5.24 (m, 1 H), 5.54 (s, 2 H), 6.90 (dd, J = 8.65, 20 2.20 Hz, 1 H), 6.97 - 7.08 (m, 2 H), 7.11 (d, J = 2.35 Hz, 1 H), 7.17 - 7.35 (m, 5 H), 7.42 (dd, J = 12.31, 8.50 Hz, 1 H), 7.62 (d, J = 8.79 Hz, 1 H), 7.68 - 7.78 (m, 1 H). Example 21 25 1-benzyl-3-(3,4-difluorobenzylcarbamoyl)-2-isopropyl-1H-indol-6-yl Dimethylcarbamate (Compound 21). General Procedure B. To a solution of 1-benzyl-N-(3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3 carboxamide (Compound 8, 18mg, 0.041mol) in pyridine (1 ml) was added dimethylcarbamyl chloride (40 pl, 0.41 mmol) and stirred at room temperature 30 overnight. The reaction was quenched with water, extracted with ethyl acetate. The combined organic layer was washed with water, brine, dried over Na 2

SO

4

,

WO 2008/089015 PCT/US2008/050695 39 and concentrated in vacuo. The residue was purified by chromatography on silica gel (0--+50% EtOAc-hexanes) to yield the title compound as a white solid (17 mg, 82%). 'H NMR (300 MHz, CD 3 0D) 8 ppm 1.32 (d, J = 7.04 Hz, 6 H), 2.96 (s, 3 H), 5 3.09 (s, 3 H), 3.37 - 3.55 (m, 1 H), 4.58 (s, 2 H), 5.48 (s, 2 H), 6.87 (dd, J = 8.65, 1.91 Hz, 1 H), 6.91 - 6.99 (m, 2 H),7.02 (d, J= 2.05 Hz, 1 H), 7.16 - 7.39 (m, 6 H), 7.58 (d, J = 8.79 Hz, 1 H). Example 22 10 1-Benzyl-3-(3,4-difluorobenzylcarbamoyl)-2-isopropyl-1H-indol-6-yl Pivalate (Compound 22). Following General Procedure B, 1-benzyl-N-(3,4 difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 18mg, 0.04 1mol) in pyridine (1 ml) was reacted with pivaloyl chloride (5.1 pd, 0.41 mmol) to yield the title compound (16 mg, 74%). 15 'H NMR (300 MHz, CD 3 0D) 8 ppm 1.25 - 1.40 (m, 15 H), 3.34 - 3.55 (m, 1 H), 4.58 (d, J = 5.86 Hz, 2 H), 5.49 (s, 2 H), 6.73 - 6.88 (m, 1 H), 6.89 - 6.99 (m, 2 H), 7.00 (d, J = 1.76 Hz, 1 H), 7.14 - 7.41 (m, 6 H), 7.60 (d, J = 8.50 Hz, 1 H). Example 23 20 1-Benzyl-3-(3,4-difluorobenzylcarbamoyl)-2-isopropyl-1H-indol-6-yl Acetate (Compound 23). Following General Procedure B, 1-benzyl-N-(3,4 difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 7mg, 0.016mol) in pyridine (1 ml) was reacted with acetyl chloride (1.0 pil, 0.16 mmol) to yield the title compound (8 mg, 100%). 25 'H NMR (300 MHz, CDCl 3 ) S ppm 1.37 (d, J 7.04 Hz, 6 H), 2.26 (s, 3 H), 3.54 - 3.76 (m, 1 H), 4.66 (d, J= 6.16 Hz, 2 H), 5.41 (s, 2 H), 6.28 (t, J = 6.01 Hz, 1 H), 6.81 - 7.01 (m, 4 H), 7.06 - 7.19 (m, 2 H), 7.18 - 7.35 (m, 4 H), 7.61 (d, J = 9.09 Hz, 1 H). 30 Example 24 WO 2008/089015 PCT/US2008/050695 40 1-Benzyl-3-(3,4-difluorobenzylcarbamoyl)-2-isopropyl-1H-indol-6-yl Propionate (Compound 24). Following General Procedure B, 1 -benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 7mg, 0.0 16mol) in pyridine (1 ml) was reacted with propionyl 5 chloride (1.4 pd, 0.16 mmol) to yield the title compound (8 mg, 100%). 'H NMR (300 MHz, CDC1 3 ) 8 ppm 1.23 (t, J = 7.48 Hz, 3 H), 1.37 (d, J = 7.33 Hz, 6 H), 2.55 (q, J = 7.43 Hz, 2 H), 3.53 - 3.73 (m, 1 H), 4.66 (d, J = 5.86 Hz, 2 H), 5.41 (s, 2 H), 6.30 (t, J = 5.72 Hz, 1 H), 6.83 - 7.00 (m, 4 H), 7.06 - 7.18 (m, 2 H), 7.18 - 7.35 (m, 4 H), 7.60 (d, J= 8.50 Hz, 1 H). 10 Example 25 1-Benzyl-3-(3,4-difluorobenzylcarbamoyl)-2-isopropyl-1H-indol-6-yl Isobutyrate (Compound 25). Following General Procedure B, 1-benzyl-N (3,4-difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide 15 (Compound 8, 9mg, 0.021mol) in pyridine (1 ml) was reacted with isobutyryl chloride (4.1 pl, 0.21 mmol) to yield the title compound (8 mg, 80%). 'H NMR (300 MHz, CDCl 3 ) 8 ppm 1.13 - 1.42 (m, 12 H), 2.47 - 2.85 (m, 1 H), 3.50 - 3.74 (m, 1 H), 4.66 (d, J = 6.16 Hz, 2 H), 5.41 (s, 2 H), 6.20 - 6.44 (m, 1 H), 6.74 - 7.00 (m, 4 H), 7.07 - 7.18 (m, 2 H), 7.17 - 7.35 (m, 4 H), 7.60 (d, J 20 8.50 Hz, 1 H). Example 26 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-(methoxymethoxy)-1H indole-3-carboxamide (Compound 26). To a solution of 1-benzyl-N-(3,4 25 difluorobenzyl)-6-hydroxy-2-isopropyl-1H-indole-3-carboxamide (Compound 8, 39 mg, 0.090 mmol) in CH 2 Cl 2 (2.0 ml) was added i-Pr 2 NEt (47 p1l, 0.27 mmol) and MOMCl (35 pl, 0.45 mmol). The reaction was stirred at room temperature for 4 h, and was purified directly by PTLC on silica gel (30% EtOAc-hexanes) to yield the title compound as a white solid. 30 'H NMR (300 MHz, CHLOROFORM-d) 6 ppm 1.37 (d, J = 7.0 Hz, 6 H), 3.42 (s, 3 H), 3.59 - 3.78 (m, 1 H), 4.66 (d, J= 5.9 Hz, 2 H), 5.10 (s, 2 H), 5.40 (s, 2 WO 2008/089015 PCT/US2008/050695 41 H), 6.29 (t, J= 5.7 Hz, 1 H), 6.85 (d, J= 2.1 Hz, 1 H), 6.89 - 7.01 (m, 3 H), 7.10 - 7.17 (m, 2 H), 7.20 - 7.34 (m, 4 H), 7.52 (d, J = 8.5 Hz, 1 H). Example 27 5 1-Benzyl-N-(3,4-difluorobenzyl)-2-isopropyl-6-(tetrahydrofuran-2-yloxy) 1H-indole-3-carboxamide (Compound 27). To a solution of 1-benzyl-N-(3,4 difluorobenzyl)-6-hydroxy-2-isopropyl- 1 H-indole-3-carboxamide (Compound 8, 39 mg, 0.090 mmol) in CH 2 Cl 2 (2.0 ml) was added 2,3-dihydrofuran (68 pl, 0.90 mmol) and catalytic amount of PPTS. The reaction was stirred at room 10 temperature for 4 h, and was purified directly by PTLC on silica gel (30% EtOAc-hexanes) to yield the title compound as a white solid. 'H NMR (300 MHz, CHLOROFORM-d) 8 ppm 1.36 (d, J = 7.3 Hz, 6 H), 1.85 - 1.98 (m, 1 H), 2.01 - 2.19 (m, 3 H), 3.58 - 3.73 (m, 1 H), 3.85 - 3.95 (m, 1 H), 3.96 - 4.07 (m, 1 H), 4.66 (d, J = 5.9 Hz, 2 H), 5.40 (s, 2 H), 5.70 (d, J = 4.7 15 Hz, 1 H), 6.29 (t, J= 5.7 Hz, 1 H), 6.86 (d, J= 2.1 Hz, 1 H), 6.89 - 6.99 (m, 3 H), 7.11 - 7.17 (m, 2 H), 7.19 - 7.32 (m, 4 H), 7.51 (d, J= 8.5 Hz, 1 H). Scheme 4 S TsCI 9-o Nal OH 0-eto1eX OH:>- Pyridine O acetone 20 28 29 Example 28 Tetrahydrofuran-3-yl 4-methylbenzenesulfonate (Compound 28). To a solution of tetrahydrofuran-3-ol (500 mg, 5.67 mmol) in pyridine (10 ml) at 0 C 25 was added 4-methylbenzene-1-sulfonyl chloride (1.08 g, 5.67 mmol). The reaction was stirred at room temperature overnight. The reaction was quenched with water, extracted with ethyl acetate. The organic layer was washed with water, brine, dried over Na 2

SO

4 and concentrated in vacuo to yield crude oil (1.2 g).

WO 2008/089015 PCT/US2008/050695 42 'H NMR (300 MHz, CDCl 3 ) 8 ppm 1.91 - 2.23 (m, 2 H), 3.61 - 4.05 (m, 4 H), 4.95 - 5.24 (m, 1 H), 7.36 (d, J = 7.92 Hz, 2 H), 7.80 (d, J = 8.50 Hz, 2 H). Example 29 5 3-lodotetrahydrofuran (Compound 29). To a solution of crude tetrahydrofuran-3-yl 4-methylbenzenesulfonate (Compound 28, 1.2 g, 4.96 mmol) in dry acetone (50 ml) was added NaI (1.1 g, 7.44 mmol). The reacted was heated at 60 C for 2 days. The mixture was diluted with water and extracted with diethyl ether. The organic layer was washed with water, brine, 10 dried over Na 2

SO

4 and concentrated in vacuo to yield crude oil which was used directly without purification. 'H NMR (300 MHz, CDCl 3 ) 8 ppm 2.23 - 2.55 (m, 2 H), 3.81 - 4.08 (m, 3 H), 4.08 - 4.43 (m, 2 H). The foregoing description details specific methods and compositions that 15 can be employed to practice the present invention, and represents the best mode contemplated. Thus, however detailed the foregoing may appear in text, it should not be construed as limiting the overall scope hereof; rather, the ambit of the present invention was to be governed only by the lawful construction of the appended claims. In particular, the present invention includes a 6-substituted 20 indole-3-carboxylic acid-N- arylmethyl amide having sphingosine-1-phosphate antagonist activity wherein the 6-substituent is represented by the formula -A I -(X')r-A2-B 25 wherein X, is 0; r is 0 or 1;

A

2 is absent or is (CH 2 )v, wherein v is 1 or 2; B is OR 6 or NR 8

R

9 , wherein R 6 , R 8 and RW are methyl; or B is CR' 0 =NO R"R'O wherein R1 0 is H and WO 2008/089015 PCT/US2008/050695 43 R" is methyl or i-butyl; or B is CONRSR 9 , wherein R 8 and R 9 are selected from the group consisting of H, methyl, ethyl and propyl or R8 and R 9 , together with N, form a 5-membered ring; or B is OR 6 , wherein R 6 is H; or B is COR 0 , wherein R1 0 is methyl. 5

Claims (20)

1. Compounds represented by the formula I having sphingosine-1 5 phosphate receptor agonist and or antagonist biological activity: R1 /N ($0R3 Y(R 4 )n-A-(X)q Z 1-(X 1 )r'BA2B Formula I wherein: 10 R1 R 2 , R 3 and R 4 are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 carbons and 1 or 2 triple bonds, carbocyclic hydrocarbon groups having 15 from 3 to 20 carbon atoms, heterocyclic groups having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, halo, CI to C 1 2 haloalkyl, hydroxyl, C 1 to C 1 2 alkoxy, C 3 to C 20 arylalkyloxy, C 1 to C 1 2 alkylcarbonyl, formyl, oxycarbonyl, carboxy, CI to C 12 alkyl carboxylate, C 1 to C 1 2 alkyl amide, aminocarbonyl, amino, cyano, diazo, 20 nitro, thio, sulfoxyl, and sulfonyl groups; X and X 1 are independently selected from the group consisting of NR 5 , 0 and S; R 5 is hydrogen, an alkyl group of 1 to 10 carbons, a cycloalkyl group of 5 25 to 10 carbons, phenyl or lower alkylphenyl; Y is a carbocyclic aryl or heterocyclic aryl group wherein said carbocylic aryl comprises from 6 to 20 atoms and said heterocyclic aryl comprises from 2 to 20 carbon atoms and from 1 to 5 heteroatoms selected from the WO 2008/089015 PCT/US2008/050695 45 group consisting of nitrogen, oxygen and sulfur, and wherein said aryl may be bonded to A at any position; Z is O or S; n is 0 or an integer of from 1 to 5; 5 o is 0 or an integer of from 1 to 3; p is 0 or an integer of from I to 3; q is 0 or 1; r is 0 or 1; A, A' and A 2 are independently selected from the group consisting of 10 (CH 2 )v wherein v is 0 or an integer of from 1 to 12, branched chain alkyl having 3 to 12 carbons, cycloalkyl having 3 to 12 carbons, alkenyl having 2 to 10 carbons and 1-3 double bonds and alkynyl having 2 to 10 carbons and 1 to 3 triple bonds; B is selected from the group consisting of hydrogen, OR 6 , COOR 7 , 15 NRR 9 , CONRR 9 , COR' 0 , CH=NOR", CH=NNR 2 R" wherein R 6, R , R10 and R" are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 carbons and 1 or 2 triple bonds, a carbocyclic hydrocarbon 20 group having from 3 to 20 carbon atoms, a heterocyclic group having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, R', R 9 , R 2 and R" 3 are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl 25 having 2 to 6 carbons and 1 or 2 triple bonds, a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, a heterocyclic group having up to WO 2008/089015 PCT/US2008/050695 46 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, or R and R 9 and/or R1 2 and R 3 , together, can form a divalent carbon radical of 2 to 5 carbons to form a heterocyclic ring with nitrogen, wherein any of R 6 , R , R , R 9 , R' 0 , R", R' or R may be substituted with 5 one or more halogen, hydroxy, alkyloxy, cyano, nitro, mercapto or thiol radical; provided however, when v is 0, and r is 0, B is not hydrogen; or B is a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, or a heterocyclic group having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, and wherein when said B is a 10 carbocyclic or heterocyclic group B may be bonded to A2 at any position, or a pharmaceutically acceptable salt of said compound.

2. The compound of claim 1 wherein Z is 0.

3. The compound of claim 2 wherein Y is a phenyl group or a pyridyl 15 group.

4. The compound of claim 3 wherein A is CH 2 .

5. The compound of claim 4 wherein X is NH. 20

6. The compound of claim 5 wherein n is 0 or an integer of 1 or 2 and R 4 is fluoro.

7. The compound of claim 6 wherein R' is i-propyl. 25

8. The compound of claim 7 wherein R3 is selected from the group consisting of phenyl, which may be substituted with one or two flouro groups, and pyridyl. 30

9. The compound of claim 8 wherein p is 0. WO 2008/089015 PCT/US2008/050695 47

10. The compound of claim 9 wherein A' and A2 are absent.

11. The compound of claim 10 wherein B is OR 6 . 5

12. The compound of claim 10 wherein B is COOR 7 .

13. The compound of claim 10 wherein X1 is 0, r is 1, A' is absent, A2 is (CH 2 )v, wherein v is 1 or 2, and B is OR 6 or NR 8 R 9 . 10

14. The compound of claim 13 wherien R 6 , R 8 and R 9 are methyl.

15. The compound of claim 10 wherein B is CR 10 =NOR"R1 0 wherein R' 0 is H and R" is methyl or i-butyl. 15

16. The compound of claim 10 wherein B is CONRR 9 wherein R 8 and R 9 are selected from the group consisting of H, methyl, ethyl and propyl, or R 8 and R 9 , together with N, form a 5-member ring. 20

17. The compound of claim 10 wherein A' is absent, r is 0, A2 is CH 2 and B is OR6, wherein R6 is H.

18. The compound of claim 10 wherein A' is absent, X is 0, r is l and B is COR' 0 wherein R1 0 is methyl. 25

19. A 6-substituted indole-3-carboxylic acid-N- arylmethyl amide having spingosine- 1-phosphate antagonist activity wherein the 6-substituent is represented by the formula (X)r-A 2-B 30 wherein X' is 0; r is 0 or 1; A 2 is absent or is (CH 2 )v, wherein v is 1 or 2; B is OR6 or NR R 9 , wherein R 6, R8 and R9 are methyl; or B is CR=NO R"R 0 wherein R1 0 is H and 35 R" is methyl or i-butyl; or WO 2008/089015 PCT/US2008/050695 48 B is CONR 8 R 9 , wherein R 8 and R 9 are selected from the group consisting of H, methyl, ethyl and propyl or R 8 and R 9 , together with N, form a 5-membered ring; or B is OR6, wherein R6 is H; or B is COR' 0 , wherein R1 0 is methyl. 5

20. A method of treating a disease or condition selected from the group consisting of glaucoma, dry eye, angiogenesis, cardiovascular conditions and diseases, and wound healing, which comprises administering to a patient in need 10 thereof a compound having sphingosine-l-phosphate receptor agonist and or antagonist biological activity represented by the general formula I: R1 / N R3, Y(R 4 ),A-(X)q / NgR3 ZR A1(X )r'2 B R~) 21P Formula I 15 wherein: R1 R2, R3 and R 4 are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 20 carbons and 1 or 2 triple bonds, carbocyclic hydrocarbon groups having from 3 to 20 carbon atoms, heterocyclic groups having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, halo, CI to C 12 haloalkyl, hydroxyl, CI to C 12 alkoxy, C 3 to C 2 0 arylalkyloxy, C 1 to C 1 2 alkylcarbonyl, formyl, oxycarbonyl, carboxy, CI to C 1 2 alkyl 25 carboxylate, CI to C 12 alkyl amide, aminocarbonyl, amino, cyano, diazo, nitro, thio, sulfoxyl, and sulfonyl groups; WO 2008/089015 PCT/US2008/050695 49 X and X 1 are independently selected from the group consisting of NR 5 , 0 and S; R 5 is hydrogen, an alkyl group of 1 to 10 carbons, a cycloalkyl group of 5 to 10 carbons, phenyl or lower alkylphenyl; 5 Y is a carbocyclic aryl or heterocyclic aryl group wherein said carbocylic aryl comprises from 6 to 20 atoms and said heterocyclic aryl comprises from 2 to 20 carbon atoms and from 1 to 5 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and wherein said aryl may be bonded to A at any position; 10 Z is O or S; n is 0 or an integer of from 1 to 5; o is 0 or an integer of from 1 to 3; p is 0 or an integer of from 1 to 3; q is 0 or 1; 15 r is 0 or 1; A, A' and A 2 are independently selected from the group consisting of (CH 2 )v wherein v is 0 or an integer of from 1 to 12, branched chain alkyl having 3 to 12 carbons, cycloalkyl having 3 to 12 carbons, alkenyl having 2 to 10 carbons and 1-3 double bonds and alkynyl having 2 to 10 carbons 20 and 1 to 3 triple bonds; B is selected from the group consisting of hydrogen, OR 6 , COOR 7 , NR 8 R 9 , CONR R9, COR'", CH=NOR", CH=NNR1 2 R1 3 wherein R 6 , R 7 , R1 0 and R' are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 25 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl having 2 to 6 carbons and 1 or 2 triple bonds, a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, a heterocyclic group having up to WO 2008/089015 PCT/US2008/050695 50 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, R', R' , R1 2 and R1 3 are are independently selected from the group consisting of hydrogen, straight or branched chain alkyl having 1 to 12 carbons, alkenyl having 2 to 6 carbons and 1 or 2 double bonds, alkynyl 5 having 2 to 6 carbons and 1 or 2 triple bonds, a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, a heterocyclic group having up to 20 carbon atoms and at least one of oxygen, nitrogen and/or sulfur in the ring, or R8 and R 9 and/or R1 2 and R 3 , together, can form a divalent carbon radical of 2 to 5 carbons to form a heterocyclic ring with nitrogen, 10 711 12 13 10 wherein any of R 6 , R 7 , R , R 9 , R1, R", R or R may be substituted with one or more halogen, hydroxy, alkyloxy, cyano, nitro, mercapto or thiol radical; provided however, when v is 0, and r is 0, B is not hydrogen; or B is a carbocyclic hydrocarbon group having from 3 to 20 carbon atoms, or a heterocyclic group having up to 20 carbon atoms and at least one of 15 oxygen, nitrogen and/or sulfur in the ring, and wherein when said B is a carbocyclic or heterocyclic group B may be bonded to A2 at any position, or a pharmaceutically acceptable salt of said compound. 20