LTB.sub.4 Synthesis Inhibitors

BACKGROUND OF THE INVENTION

SUMMARY OF THE INVENTION

DETAILED DESCRIPTION

EXAMPLE 1 ##STR8## The above acid chloride was prepared from terphthalic acid by reacting 0.5 g (3 mmoles) of terphthalic acid with 2 cc of [COCl].sub.2 (23.6 mmoles) in 10 cc of benzene and with one drop of dimethylformamide. The reagents were mixed and warmed to 60.degree. C. for twenty-four hours. The reaction mixture was cooled to room temperature and the volatile components were removed in vacuo to give the above compound as a pale yellow solid.

EXAMPLE 2

EXAMPLE 3 ##STR9## The above compound was prepared by reacting 3 mmoles of the acid chloride product from Example 1 with 0.8 g (3 mmoles) of the TMS-acetylene product from Example 2. The acid chloride and the TMS-acetylene product were dissolved in 10 cc of dichloromethane and cooled to 0.degree. C. To the reaction mixture was added 0.8 g (6 mmoles) of aluminum chloride (AlCl.sub.3) in small portions over ten minutes. The reaction mixture was stirred for about 1.5 hours at 0.degree. C. The reaction was quenched with ice and the mixture was extracted three times with diethyl ether. The extracts were combined and washed once with water and once with brine (saturated sodium bicarbonate solution) and dried over magnesium sulfate to yield 0.29 g of the above product.

EXAMPLE 4 ##STR10## The above compound was prepared by mixing 3 g of m-iodobenzoic acid (12.1 mmoles) and 3.12 g (15.0 mmoles) of an acetylene derivative of the formula ##STR11## with 0.085 g (0.121 mmoles) of a palladium catalyst, Pd(PPh.sub.3).sub.2 Cl.sub.2 in 30 cc of Et.sub.3 N. The reaction vessel was purged with carbon monoxide. The reaction mixture was heated under a carbon monoxide atmosphere (atmospheric pressure, balloon) in an oil bath at 80.degree. C. for two hours. The reaction mixture was cooled to room temperature. The volatile components were removed in vacuo and the residue was taken up in 5% hydrochloric acid and extracted with diethyl ether. The diethyl ether was washed once with 10% hydrochloric acid, twice with water, and once with a brine solution and dried over magnesium sulfate. The solvent was removed yielding 4.58 gm of the product. Chromatography on silica gel afforded 3.2 g of yellow solid which was triturated with cold hexane to give a tan solid of 2.97 g.

EXAMPLE 5 ##STR12## The above compound was prepared by mixing one gram (4.03 mmoles) of m-iodobenzoic acid and 0.83 g (5 mmoles) of an acetylene derivative of the formula ##STR13## along with 0.028 g (0.04 mmoles) of the palladium catalyst used in Example 4. The reagents were mixed in 10 cc of triethylamine. The reaction vessel was purged with carbon monoxide. The reaction was performed under a carbon monoxide atmosphere (atmospheric pressure, balloon). The reaction mixture was heated in an oil bath at 80.degree. C. for two hours. The reaction mixture was cooled to room temperature. The volatile components were removed in vacuo and the residue was taken up in 5% hydrochloric acid (HCl) and extracted once with diethyl ether. The diethyl ether extract was washed once with 10% HCl, twice with water and once with brine (NaCl) and dried over magnesium sulfate. The solvent was removed yielding a red yellow gum. After purification by silica gel chromatography 0.51 g of the product having the above structure was produced.

EXAMPLE 6 ##STR14## A solution of l g (4.8 mmoles) of an acetylene having the formula

EXAMPLE 7 ##STR15## A solution of 10 g (52.4 mmoles) of a bromo-furanoic acid of the formula ##STR16## in 100 cc of methanol was prepared and cooled to 0.degree. C. using an ice bath. 20 ml of concentrated sulfuric acid was added dropwise over 15 minutes. The ice bath was removed and the reaction mixture stirred for eighteen hours at room temperature overnight. The reaction mixture was poured onto 500 ml of water and extracted twice with diethyl ether. The extracts were combined and washed once with water, once with NaHCO.sub.3, once again with water and brine and dried over magnesium sulfate. The solvent was removed to yield 6.77 g of a pale yellow solid.

EXAMPLE 8 ##STR17## The above compound was prepared by reacting 2.02 g (9.7 mmoles) of the acetylene compound of the structure ##STR18## in 15 ml of THF at 0.degree. C. To the solution was added 6.5 ml (10.4 mmoles) of 1.6 molar n-butylithium. The reaction mixture was stirred for 25 minutes at 0.degree. C. To the cooled milky solution was added 3 ml (11.1 mmoles) of ClSnBu.sub.3 in a dropwise manner. The solution cleared to a pale yellow. The reaction was allowed to warm to room temperature. The reaction mixture was diluted with dichloromethane and washed with water and brine. The reaction mixture was dried over sodium sulfate, filtered and stripped to yield 5.13 g of the above product as a pale yellow oil.

EXAMPLE 9 ##STR19## The above compound was prepared by reacting 0.21 g (1.01 mmoles) of the product from Example 7 and 0.5 g (1.01 mmoles) of the acetylene product from Example 8 in 5 cc of toluene. The palladium catalyst of Example 4, 7 mg (0.01 mmoles) was added to the reaction mixture. The reaction mixture was degassed and purged three times with carbon monoxide. The reaction mixture was heated 100.degree. C. using an oil bath and stirred for eight hours at 100.degree. C. The toluene solution was treated with saturated aqueous potassium fluoride solution for 1/2 hour. The mixture was then poured into diethyl ether and extracted twice with water and dried over magnesium sulfate. After filtration and removal of the solvent, a brown gummy solid was obtained. Following chromatography on silica gel the above product was obtained, 0.08 g (0.22 mmoles).

EXAMPLE 10 ##STR20## The above compound was prepared by forming a solution of 2.5 gms. (13.1 mmoles) of a bromo-furanoic acid in 25 ml of tetrahydrofuran (THF) which was cooled to -78.degree. C. To the solution was added a 1.6 molar solution of n-butyl lithium in hexane (17.5 ml, 28 mmoles) which was stirred at -78.degree. for one hour. Dimethylformaide (DMF) was added in an amount of 2.4 ml (30 mmoles). The solution was allowed to warm to room temperature. The reaction mixture was quenched with water and acidified with 10% hydrochloric acid. The resultant reaction mixture was extracted twice with ethyl acetate, the combined extracts were washed twice with water and once with brine and subsequently dried over magnesium sulfate. An orange solid was obtained after removal of the solvent in vacuo. Following chromatography on silica gel 0.73 gms. of the compound of the above formula was obtained.

EXAMPLE 11 ##STR21## The above compound was prepared by forming a solution of 1.2 gms. (6 mmoles) of an acetylene of the formula H.dbd.(CH.sub.2).sub.12 CH.sub.3 in 25 ml. THF which contained a catalytic amount (20 mg.) of triphenylmethane. The solution was cooled to -50.degree. C. and then treated with 3.75 ml. (6 mmoles) of n-BuLi until the red color of the triphenylmethane anion persisted. A few drops of the acetylene compound was added until the color disappeared. An amount of 0.42 gms. of the product formed in Example 10 in 10 ml. THF was added drop-wise to the solution. The mixture was warmed to 0.degree. C. over one-half hour. The mixture was quenched with water and acidified with 10% hydrochloric acid. The aqueous phase was extracted twice with ethyl acetate. The extracts were combined and washed twice with water and once with brine and were dried over magnesium sulfate. Chromatography on silica gel yielded 0.80 gms. of a pale yellow solid.

EXAMPLE 12 ##STR22## The compound was prepared by reacting 0.145 g. (4.2 mmoles) of the product from example 11 in acetone (25 ml) and adding 1.5 g. of activated MnO.sub.2 portionwise over five minutes. The reaction mixture was stirred for 24 hours at room temperature. The reaction mixture was poured into 10% hydrochloric acid and extracted with ethyl acetate. The extract was washed once with water and dried over magnesium sulfate. After the solvent was removed in vacuo a white solid remained. Following chromatography on silica gel, 60 mg. of a white solid was recovered.

EXAMPLE 13 ##STR23## The above compound was prepared by forming a solution of 0.83 gms. (4 mmoles) of an acetylene of the formula H--C.dbd.C--(CH.sub.2).sub.12 CH.sub.3 in 25 ml. THF containing a catalytic amount (20 mgs.) of triphenyl methane. The solution was cooled to -50.degree. C., then treated with 2.5 ml. of a 1.6 molar solution (4 mmoles) of n-BuLi in hexane until the red color of the triphenylmethane anion persisted. A few drops of the acetylene was added until the color disappeared. The resultant lithium acetylide preparation was cooled to -78.degree. C. A solution (6 mmoles) of a diacid chloride of the formula ##STR24## was cooled to -78.degree. C. and the -78.degree. C. solution of lithium acetylide was added dropwise via a cannula. The reaction was stirred for 10 minutes and quenched with water and warmed to room temperature. The reaction mixture was poured into water and acidified with acetic acid (HOAc). The aqueous solution was extracted twice with ethyl acetate and the extracts were washed twice with water, once with brine and dried over magnesium sulfate. Following chromatography on silica gel 0.95 gms. of a product of the above formula was recovered.

EXAMPLE 14 ##STR25## The compound of the above formula was prepared in the following manner. An acetylene solution was prepared by dissolving 0.83 gms. (4 mmoles) of an acetylene of the formula H--C.dbd.C--(CH.sub.2).sub.12 CH.sub.3 in 25 ml. of THF and 20 mgs. of triphenylmethane. The solution was cooled to -50.degree. C. and treated with n-BuLi until the red color of the triphenylmethane anion persisted. A few drops of the acetylene was added until the color disappeared. An aldehyde of the following formula ##STR26## in an amount of 0.36 gms. (2 mmoles) was added dropwise to the solution and the mixture was warmed to 0.degree. C. The reaction mixture was quenched with water and acidified with 10% hydrochloric acid. The aqueous phase was extracted twice with ethyl acetate and the extracts were washed twice with water, once with brine and dried over magnesium sulfate. Upon chromatography over silica gel 0.55 gms. (1.42 mmoles) of a product of the above formula was recovered.

EXAMPLE 15 ##STR27## The above three compounds were prepared in the following manner. An acid chloride of the structure ##STR28## was prepared in a manner as recited in Example 1 in an amount of 15.05 mmoles was dissolved in 50 ml. of dichloromethane and cooled to 0.degree. C. To the solution was added 3.2 gms. (15 mmoles) of an acetylene of the formula H--C.dbd.C--(CH.sub.2).sub.12 CH.sub.3. 2 gms. of aluminum chloride was added portionwise over one-half hour and the reaction mixture was stirred at 0.degree. C. for one hour. The reaction mixture was quenched with ice, diluted with ether and washed twice with water and dried over magnesium sulfate. Following separation by chromatography on silica gel 42 mg. and 30 mg of the isomers of the above formulae 15a and 15b were obtained and 23 mg. of the isomer of the above formula 15c was obtained.

EXAMPLE 16 ##STR29## A compound of the above structure was prepared by forming a solution of an acetylene of the formula H--C.dbd.C--(CH.sub.2).sub.12 CH.sub.3 in 25 ml. of THF and 20 mgs. of triphenylmethane. The solution was cooled to -50.degree. C. and treated with n-BuLi until the red color of the triphenylmethane anion persisted. A few drops of acetylene was added until the color disappeared. An aldehyde of the formula ##STR30## in an amount of 0.33 gms. (2 mmoles) in 5 ml. of THF was added dropwise to the solution and the mixture was warmed to 0.degree. C. The reaction mixture was quenched with water and acidified with 10% hydrochloric acid. The aqueous phase was extracted twice with ethyl acetate and the extracts were washed twice with water, once with brine and dried over magnesium sulfate. Upon separation by chromatography on silica gel, 0.447 gms. (1.2 mmoles) of the product of the above formula was recovered as a white solid.

EXAMPLE 17 ##STR31## The compound of the above formula was prepared in the following manner. 0.1 gms. (0.27 mmoles) of the compound prepared in Example 16 was dissolved in 10 cc. of acetone. Activated manganese dioxide (1 gm.) was added portionwise. The reaction mixture was stirred for 24 hours. The crude reaction mixture was filtered through celite and the solvent evaporated. Following chromatography over silica gel 0.55 gms. of a product of the above formula was produced as a white solid.

EXAMPLE 18 ##STR32## The above compound was prepared in the following manner. Decadiyne in the amount of 6.71 gms. (15 mmoles) was dissolved in 200 ml. of THF, along with 50 mgs. of triphenylmethane. The mixture was cooled to -50.degree. C. and 31.3 ml. (50 mmoles) of 1.6 molar n-BuLi in hexane was added until the red color of the triphenylmethane anion persisted. Additional decadiyne was added until the red color disappeared. The reaction mixture was warmed to -20.degree. C. and stirred for one-half hour. The reaction mixture was cooled to -40.degree. C. and 9.9 gms. (50 mmoles) of iodohexane was added dropwise. Following the addition of the iodohexane, 50 ml. of HMPA was added dropwise and the reaction mixture was stirred and allowed to warm to room temperature. The reaction mixture was quenched with water and poured into hexane and washed with four washings of water, one with brine and dried over magnesium sulfate. The crude product contained the diyne of the formula H--C.dbd.C--(CH.sub.2 ).sub.6 --C.dbd.C--C.sub.5 H.sub.11.

EXAMPLE 19 ##STR34## The compound of the above formula was prepared in the following manner. A solution was prepared by dissolving 0.125 gms. (0.35 mmoles) of the product from Example 18 in 15 ml. of acetone. To the solution was added 1 gm. of manganese dioxide (MnO.sub.2). The manganese dioxide was added portionwise over a period of 15 minutes. The reaction mixture was stirred for 24 hours at room temperature. The reaction mixture was poured into 10% hydrochloric acid and extracted with ethyl acetate. The extract was washed once with water and dried over magnesium sulfate. The solvent was removed in vacuo and the product was isolated using chromatography on silica gel to provide 0.90 gms. of a product of the above formula.

EXAMPLE 20 ##STR35## A compound of the above formula was prepared in the following manner. 10 mgs. (0.028 mmoles) of the compound prepared in Example 19 was mixed with 2.0 ml. of hexane and 1 mg. of a Lindlar catalyst and 0.01 ml. of a quinoline. The reagents were mixed and the reaction vessel was degassed 3 times with hydrogen delivered from a balloon. The reaction mixture was monitored closely via TLC and stirred at room temperature. The reaction vessel was evacuated and purged three times with argon. The reaction mixture was then filtered through celite. The filtrate was washed once with 10% hydrochloric acid, once with water, once with saturated sodium bicarbonate, once with saturated sodium chloride, and dried over magnesium sulfate. Following separation by chromatography 0.008 gms. of a product of the above formula was recovered as a mixture of E and Z isomers about the enone double bond.

EXAMPLE 21 ##STR36## A compound of the above structure was prepared in the following manner. To a solution of 0.1 gms. of the product from Example 18 (0.282 mmoles) in 1 ml. of DMF was added 0.1 gms. (1.5 mmoles) of imidazole, followed by 0.19 gms. (0.7 mmoles) of t-butyldiphenylsilyl chloride (TBDPS-Cl) at room temperature under argon. The solution was stirred for 4 hours at room temperature. The reaction mixture was diluted with 200 ml. of diethyl ether and washed 3 times with water (30 ml. each), once with brine and dried over magnesium sulfate. Removal of the solvent yielded an oil which upon separation by chromatography on silica gel yielded 0.11 gms. of a product of the formula ##STR37## This product, 0.11 g (0.13 mmoles) was mixed with 10 ml. of hexane, 5 mgs. of a Lindlar catalyst and 0.05 ml of quinoline. The reaction mixture was stirred at room temperature under hydrogen and after 1 hour TLC indicated the reaction was complete. The reaction vessel was evacuated and purged 3 times with argon. The reaction mixture was filtered through celite, diluted with hexane and washed once with 5% hydrochloric acid, once with water, once with brine and dried over magnesium sulfate. The resultant product produced was 0.11 gms of a product of the formula ##STR38## A solution of this product was prepared by dissolving 0.11 gms. in 10 ml. of THF, which was then treated with 1.5 ml. of 1 molar n-Bu.sub.4 NF in THF. After the starting material was consumed (via TLC), the reaction mixture was diluted with diethyl ether and washed with water and dried over magnesium sulfate. Following chromatography 0.037 gms. (0.103 mmoles) of the above identified product was recovered.

EXAMPLE 22 ##STR39## A compound of the above formula was prepared in the following manner. The compound prepared in Example 21, 0.030 gms., was dissolved in 3 ml. of acetone and 0.3 gms. of MnO.sub.2 was added portionwise over 5 minutes. The reaction mixture was stirred for 24 hours at room temperature. The reaction mixture was then poured into 10% hydrochloric acid and extracted with ethyl acetate. The extract was washed once with water and dried over magnesium sulfate. After the solvent was removed in vacuo the resultant product was separated by chromatography over silica gel, yielding 0.021 gms. of product of the above formula.

EXAMPLE 23 ##STR40## A compound of the above formula was prepared in the following manner. To a cooled (0.degree. C.) solution of 5.5 ml. (39.2 mmoles) of diisopropylamine in 50 ml. of THF was added 20.5 ml. of 1.6 molar BuLi (32.8 mmoles) to make 32.8 mmoles of lithium diisopropylamide (LDA). The reaction mixture was stirred for one-half hour at 0.degree. C. and cooled to -78.degree. C. To the mixture was added 2.1 gm (16.4 mmoles) of 2-thiophene carboxylic acid in 25 ml. THF. Additional THF was added to increase the volume to 200 ml. and the reaction mixture was stirred for one-half hour. DMF was added in an amount of 1.3 ml. (16.8 mmoles). The reaction mixture was warmed to room temperature and stirred for 11/2 hours. The reaction mixture was quenched with water and acidified with 1N hydrochloric acid and extracted with ethyl acetate. The organic extracts were combined and dried over magnesium sulfate. The resultant mixture was filtered and stripped to yield a yellow solid. Separation using chromatography on silica eluting with ethyl acetate/hexane/1% acetic acid provided 1.3 gms. of a yellow solid of the above formula. MP 160.degree.-163.degree..

EXAMPLE 24 ##STR41## The compound with the above structure was prepared in the following manner. An acetylene of the formula H--C.dbd.C--(CH.sub.2).sub.12 CH.sub.3 in an amount of 549.3 mg. (2.6 mmoles) in 15 ml. of THF was cooled to -20.degree.. To the solution was added 1.6 ml. (2.6 mmoles) of n-BuLi. The reaction was stirred for one half hour and 203.4 mg. (1.3 mmoles) of the product from Example 23 in 10 ml. of THF was added. The mixture was stirred and maintained at -20.degree. for 15 minutes and allowed to warm to 0.degree. C. and stirred for one-half hour. The reaction mixture was quenched with water and acidified with 10% hydrochloric acid and extracted with ethyl acetate. The combined organic extracts were dried over magnesium sulfate. The extract was filtered and stripped to yield a yellow oil which solidified upon standing. The solid was dissolved in ethyl acetate and filtered through silica gel eluting with 100% hexane followed by ethyl acetate in 1% acetic acid. The ethyl acetate fraction yielded 392.8 mg. of the above compound as a yellow solid having a melting point of 75.degree.-90.degree..

EXAMPLE 25 ##STR42## A compound of the above structure was formed in the following manner. 7.2 mg. (0.020 mmoles) of the compound prepared in Example 24 was dissolved in 1 ml. acetone. To the solution was added 72 mg. (0.83 mmoles) of activated manganese dioxide. The reaction mixture was stirred vigorously at room temperature overnight. The reaction mixture was poured into 10% hydrochloric acid. The aqueous phase was extracted with ethyl acetate. The organic washes were combined and dried over magnesium sulfate. The organic phase was filtered and stripped to yield 6 mg. (0.010 mmoles) of the above product as a white solid.

EXAMPLE 26 ##STR43## A compound of the above formula was prepared in the following manner. m-Iodobenzoic acid was reacted in an amount of 569 mgs. (2.29 mmoles) with 440 mg. (2.30 mmoles) of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide.HCl in 25 ml. of DMF. To the reaction mixture was added 0.35 ml. (2.51 mmoles) of Et.sub.3 N and 361 mg. (2.30 mmoles) of NH.sub.2 SO.sub.2 (C.sub.6 H.sub.5). The reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was poured into water and extracted with ethyl acetate. The organic extracts were combined and washed with brine and dried over magnesium sulfate. Following chromatography on silica gel eluting with ethyl acetate/hexane/1% acetic acid, 126 mg. of a white solid of the product with the above formula was recovered.

EXAMPLE 27 ##STR44## A compound with the above formula was prepared in the following manner. The compound from Example 26 in an amount of 77 mg. (0.20 mmoles) was combined with 49 mg. (0.24 mmoles) of an acetylene of the formula H--C.dbd.C--(CH.sub.2).sub.12 CH.sub.3 and 3 mg. (0.004 mmoles) of Pd(Ph.sub.3).sub.2 Cl.sub.2 in 2 ml. of Et.sub.3 N. The mixture was heated in an oil bath at about 80.degree. C. The reaction vessel was purged 4 times with carbon monoxide from a carbon monoxide balloon and permitted to react with stirring under carbon monoxide for 2.5 hours. The reaction mixture was then stirred at room temperature for about 60 hours. The solvent was evaporated under nitrogen. The residue was dissolved in ethyl acetate and washed with successive washes of 10% hydrochloric acid, water and brine. The organic layer was dried over magnesium sulfate, filtered and stripped to yield a brown oil. Upon chromatography on silica gel, 35 mgs. of a compound with the above formula was recovered as a brown oil.

EXAMPLE 28 ##STR45## A compound having the above formula was prepared in the following manner. A solution of m-cyanobenzaldehyde was prepared by dissolving 2.0 gms. (15.2 mmoles) of the benzaldehyde along with 2.98 gms. (45.8 mmoles) of NaN.sub.3 and 2.9 gms. (21.1 mmoles) of Et.sub.3 N.HCl, all of which were dissolved in 50 ml. of 1-methyl-2-pyrrolidinone. The reaction mixture was refluxed under argon. After 1 hour and 45 minutes the reaction mixture was cooled to room temperature and poured into 200 ml. of water and acidified with 10% hydrochloric acid. The reaction mixture was extracted with successive ethyl acetate washes. The ethyl acetate extracts were combined and washed with brine and dried over magnesium sulfate. The ethyl acetate extract was chromatographed through silica gel, yielding 0.3 gms. of the product having the above formula as a white solid.

EXAMPLE 29 ##STR46## A compound of the above formula was prepared in the following manner. A solution of 5.03 g (24.1 mmoles) of an acetylene of the formula HC.dbd.C(CH.sub.2).sub.12 CH.sub.3 was dissolved in 100 ml. THF and cooled to -30.degree. C. To the solution was added 15 ml. (24 mmoles) of a 1.6 molar n-BuLi solution which was added dropwise. The reaction mixture was stirred for 15 minutes at which time 2.1 gms. (12.0 mmoles) of the product from Example 28 dissolved in 75 ml. of THF was added dropwise. The solution was stirred and maintained at -30.degree. C. for one-half hour, then warmed to room temperature. The reaction mixture was quenched with water and acidified with 10% hydrochloric acid. The layers were separated and the organic phase was washed with brine and dried over sodium sulfate. The layer was filtered and stripped to yield a yellow solid which upon chromatography over silica gel yielded 3.75 gms. of the above product as a white solid.

EXAMPLE 30 ##STR47## The compound having the above formula was prepared in the following manner. A solution was prepared by dissolving 36.2 mgs. (0.095 mmoles) of the product from Example 29 in acetone. To the solution was added 360 mgs. (4.14 mmoles) of activated manganese dioxide. The reaction mixture was stirred vigorously at room temperature overnight. The reaction mixture was poured into 10% hydrochloric acid. The aqueous phase was extracted with ethyl acetate and the organic washes were combined and dried over magnesium sulfate. The organic phase was filtered and stripped to yield 18 mgs. of a pale yellow solid of a compound having the above formula.

ASSAY FOR LTB.sub.4 AND PGE.sub.2 PRODUCTION BY HL-60 CELLS

HUMAN SYNOVIAL FLUID PHOSPHOLIPASE A.sub.2 (HSF-PLA.sub.2) ASSAY