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United States Patent5648373
Winkler , ; et al.July 15, 1997
Title
CoA-IT and PAF inhibitors
Abstract
Coenzyme A-independent transacylase is required for the release of free arachidonic acid, and the production of arachidonic acid metabolites and platelet activation factor. Blocking of this enzyme inhibits the production of these inflammatory mediators and will be of therapeutic utility in a broad range of allergic and inflammatory diseases and disorders. Compounds are described herein which inhibit the action of CoA-IT and are therefore useful in the treatment of disease states caused thereby.
Inventors:Winkler; James David (Fort Washington, PA), Chilton, III; Floyd Harold  (Pilot Mountain, NC), Hickey; Deirdre Mary Bernadette  (Welwyn, GB2)
Assignee:SmithKline Beecham Corporation (Philadelphia, PA)
The Johns Hopkins University (Baltimore, MD)
Appl. No.:102877
Filed:August 6, 1993
Foreign Application Priority Data
Feb 11, 1992 [GB] 9202827
Current U.S. Class:514/398 514/406 514/425 514/825 514/826 514/863 514/916 514/921 514/359 514/374 514/392 514/397 
Field of Search:514/392,397,398,359,406,374,425,825,826,863,916,921
U.S. Patent Documents
3021338February 1962Bortnick
4355039October 1982Niedballa et al.
5087634February 1992Reitz et al.
5248689September 1993Girard et al.
5256695October 1993Poss
5338752August 1994Hickey et al.
Foreign Patent Documents
0 653160Jan., 1992AU
2 000 774Jan., 1979GB
9204331Mar., 1992WO
Other References
CA 116(25):255613, 19 Mar. 1992, Hickey et al. .
CA 82(25):170937W, 1975, Jorgensen. .
CA 117(3):26565, 19 Mar. 1992, Hickey et al. .
CA 117(11):111 616JA, 19 Mar. 1992, Hickey et al. .
CA 115(5):49682, 1991, Meanwell et al. .
CA 112(6):42581b, 1989, Duerr. .
CA 110(16):141549h, 1986, Schmitz et al. .
CA 105(23):20887p, 1986, Lautenschlaeger et al. .
Medline 92170539, 1991, Winkler et al. .
Medline 911521139, 1991, Winkler et al. .
CA 116(13):125547d, 1991, Suguira et al. .
CA 115(17):177545a, 1991, Uemura et al. .
Ninio et al., Regulation of the COA-Independent Transacylase in Human Neutrophils, Federation of European Biochemical Societies, Jul. 1991, pp. 138-140..~
Primary Examiner: Burn; Brain M.
Attorney, Agent or Firm:Dinner; Dara L. Venetianer; Stephen Lentz; Edward T.
Parent Case Text


This application is a continuation-in-part application of PCT application US93/01247, filed 11 Feb. 1993, which is a continuation-in-part application of U.S. Ser. Nos. 07/833,850 (now abandoned); 07/833,877 (now abandoned); 07/833,878 (now abandoned); 07/833,879 (now abandoned); 07/833,880 (now abandoned); 07/834,048 (now abandoned), all filed 11 Feb. 1992.
Claims

What is claimed is:
1. A method for treating an inflammatory component of a disease or disorder mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprises administering to a mammal in need thereof an effective amount of a compound which inhibits lipid mediator production by blocking the production, activation or action of Coenzyme A-independent transacylase (CoA-IT).

2. The method according to claim 1 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some part by the lipid inflammatory mediators.

3. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound of the formula ##STR17## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl;
n is 4 to 12;
X is 5-tetrazolyl, SO.sub.3 H, P(O)(OR.sub.2).sub.2, P(O)(OH).sub.2, or P(O)(R.sub.2)(OR.sub.2);
R.sub.2 is hydrogen or C.sub.1-4 alkyl;
R.sub.3 is independently hydrogen, C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C1-4 alkoxy;
m is a number having a value of 1 to 3;
q is a number having a value of 1 to 3;
or a pharmaceutically acceptable salt thereof.

4. The method according to claim 3 wherein the compound is
Diethyl-7-(3,4,5-triphenylimidazol-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate;
Ethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)methyl-phosphinate; or
7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptanephosphonate.

5. A compound which is
Diisopropyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptanephosph onate;
Dimethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl) heptanephosphonate;
Diethyl-6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanephosphonate ;
Diethyl-8-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanephosphonate ; or a
pharmacuetically acceptable salt thereof.

6. A pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a compound selected from the group consisting of:
Diisopropyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptanephosph onate;
Dimethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptanephosphona te;
Diethyl-6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanephosphonate ; and
Diethyl-8-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanephosphonate ; or a pharmacuetically acceptable salt thereof.

7. The method according to claim 3 or 4 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, or pyresis.

8. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound of the formula ##STR18## wherein R is hydrogen, C.sub.1-8 alkyl, C.sub.1-8 alkoxy, SC.sub.1-8 alkyl, optionally substituted phenyl, phenyl C.sub.1-4 alkyl in which the phenyl group is optionally substituted, C.sub.1-6 alkylCHO or C.sub.1-6 alkylCH(OR.sup.1)(OR.sup.2) in which each group R.sup.1 and R.sup.2 is C.sub.1-4 alkyl, or together form an ethane 1,2-diyl or propane 1,3-diyl group;
n is an integer having a value of 2 to 6;
m is an integer having a value of 0 to 6;
p is an integer having a value of 1 to 3;
R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are independently hydrogen or C.sub.1-4 alkyl;
AB is a bond, --CH.dbd.CH--, --S--, S-phenyl or O-phenyl;
X is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl;
R.sup.7 is hydrogen, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, halogen, hydroxy, or C.sub.1-4 alkoxy;
or a pharmaceutically acceptable salt thereof; provided that:
a) when X is 5-tetrazolyl, R.sup.7 is hydrogen, R is phenyl, and AB is a bond, then n+m are equal to a number greater than 6;
b) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not hydrogen;
c) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not alkyl or hydrogen;
d) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 4-hydroxy, then R is not phenyl;
e) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 4-Methoxy or is 4-hydroxy, then R is not hydrogen;
f) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 2-chloro, then R is not hydrogen;
g) when (R.sup.7)p is the same and is hydrogen, R is phenyl, n is 4, m is 0, and AB is O-phenyl then X is not CO.sub.2 --C.sub.1-6 alkyl;
h) when R is hydrogen, (R.sup.7)p is the same and is hydrogen, AB is a bond, n+m is equal to 7, than X is not CH.sub.3 O--(CH.sub.2).sub.2 --O--(CH.sub.2).sub.2 --O--C(O)--;
i) when X is CO.sub.2 --C.sub.1-6 alkyl, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not phenyl or 4-methoxyphenyl;
j) when X is CO.sub.2 --C.sub.1-6 alkyl, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 4-bromo or 4-methoxy, then R is not hydrogen;
k) when X is CO.sub.2 --C.sub.1-6 alkyl, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not 2-(4-methoxybenzyl);
l) when (R.sup.7)p is the same and is hydrogen, R is phenyl, AB is a bond n+m is equal to 10, then X is not CO.sub.2 --C.sub.1-6 alkyl;
m) when (R.sup.7)p is the same and is hydrogen, R is phenyl, n is 4, m is 0 and AB is O-phenyl, then X is not CO.sub.2 --C.sub.1-6 alkyl;
n) when AB is --S--, n is 5 or 6, then m is 1 and X is CO.sub.2 H;
or a pharmaceutically acceptable salt thereof.

9. The method according to claim 8 wherein the compound is
1-(7-Carboxyheptyl)-2-heptyl-4,5-diphenylimidazole;
1-(7-(5-Tetrazolylheptyl)-2,4,5-triphenylimidazole;
1-(10-Carboxydecyl)-2,4,5-triphenylimidazole;
4-[4-(2,4,5-triphenylimidazolyl)butyloxy]benzoic acid;
9-(1,2,4-tri-phenylimidazolyl)-2,2-dimethylnonanoic acid;
1-(8-Carboxyoctyl)-2,4,5-triphenylimidazole;
1-(7-Carboxy-heptyl)-2-(4-hydroxy-3,5-diiodophenyl)-4,5-diphenylimidazole;
Ethyl 8-(4,5-diphenylimidazol-1-yl)octanoate;
1-(7-Ethoxycarbonylheptyl)-2-methyl-4,5-diphenylimidazole; or
1-(7-Carboxyheptyl)-2-(4-hydroxyphenyl)-4,5-diphenyl-imidazole.

10. The method according to claim 8 or 9 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some part by the lipid inflammatory mediators.

11. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound of the formula ##STR19## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl;
n is an integer having a value of 4 to 12;
Y is oxygen or sulfur;
X is 5-tetrazolyl, SO.sub.3 H, P(O)(OR.sub.2).sub.2, P(O)(OH).sub.2, or P(O)(R.sub.2)(OR.sub.2);
R.sub.2 is hydrogen or C.sub.1-4 alkyl;
R.sub.3 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C1-4 alkoxy;
m is an integer having a value of 1 to 3;
q is an integer having a value of 1 to 3;
or a pharmaceutically acceptable salt thereof.

12. The method according to claim 11 wherein the compound is
Ethyl-7-(1,4,5-triphenyl-imidazol-2-yl-oxy)heptane methylphosphinate;or
Diethyl-7-(1,4,5-triphenyl-imidazol-2-yl-oxy)heptanephosphonate.

13. The method according to claim 11 or 12 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some part by the lipid inflammatory mediators.

14. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound of the formula ##STR20## wherein X is nitrogen or CR.sup.1 ;
R.sup.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl;
Y is nitrogen, N(CH.sub.2).sub.n A or C(CH.sub.2).sub.n A
Z is nitrogen, oxygen or N(CH.sub.2).sub.n A', and the dotted line indicates the optional presence of a double bond so as to form a fully unsaturated heterocyclic ring;
n is an integer having a value of 4 to 12;
A' is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl;
A is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, OH, Br, Cyano, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl;
R.sup.2 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;
m is an integer having a value of 1 to 3; provided that
a) X, Y and Z are not all at the same time, nitrogen;
b) when X is CR.sup.1, Y and Z are not both nitrogen;
c) when Y is N(CH.sub.2).sub.n A, Z is nitrogen; and
d) when Z is oxygen, Y is C(CH.sub.2).sub.n A;
e) when Y is N(CH.sub.2).sub.n A, X and Z are nitrogen, (R.sub.2).sub.m is the same and is hydrogen, and n is 6, 7, or 8 then X is not --CO.sub.2 --C.sub.1-6 alkyl;
f) when Z is oxygen, Y is C(CH.sub.2).sub.n A, n is 8, and (R.sub.2).sub.m is the same and is hydrogen, then X is not cyano;
g) when Z is N(CH.sub.2).sub.n A', X is nitrogen, Y is nitrogen, (R.sub.2).sub.m is the same and is hydrogen, and n is 7, then X is not CO.sub.2 H;
h) when Y is N(CH.sub.2).sub.n A, X and Z are nitrogen, (R.sub.2).sub.m is the same and is hydrogen, and n is 8 then X is not cyano;
or a pharmaceutically acceptable salt thereof.

15. The method according to claim 14 wherein the compound is
1-(8-Bromooctyl)-4,5-diphenyltriazole;
2-(8-Cyanooctyl)-4,5-diphenyl-triazole;
8-(3,4-Diphenylpyrazol-1-yl)octanoic acid;
2-(9-Hydroxynonyl)-4,5-diphenyl-1,2, 3-triazole
2-(7-Methoxycarbonylheptyl)-4,5-diphenyltriazole
8-(3,4-Diphenylpyrazol-1-yl)octanoic acid;
8-(4,5-Diphenylpyrazol-1-yl)octanoic acid;
2-(6-Carboxyhexyl)-4,5-triphenyltriazole; or
2-(7-Carboxyheptyl)-4,5-diphenyloxazole.

16. The method according to claim 14 or 15 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some past by the lipid inflammatory mediators.

17. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound of the Formula ##STR21## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, or optionally substituted phenyl;
n is 2 or 4 m 12;
X is cyano, CO.sub.2 H or a group hydrolysable to CO.sub.2 H;
R.sub.3 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;
q is an integer having a value of 1 to 3;
or a pharmaceutically acceptable salt thereof.

18. The method according to claim 17 wherein the compound is:
Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate;
Ethyl 6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;
9-[1-(3,4,5-Triphenyl-2-oxo-2,3-dihydroimidazolyl)]nonanoic acid;
7-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)heptanitrile;
Ethyl 6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
11-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)undecanoic acid; or
Ethyl-8-(4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanoate.

19. The method according to claims 17 or 18 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some part by the lipid inflammatory mediators.

20. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound of the Formula: ##STR22## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, or optionally substituted phenyl;
n is 4 to 12;
Y is oxygen or sulfur;
X is CO.sub.2 H or a group hydrolysable to CO.sub.2 H;
R.sub.3 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;
q is an integer having a value of 1 to 3;
or a pharmaceutically acceptable salt thereof.

21. The method according to claim 20 wherein the compound is
Ethyl 5-(1,4,5-triphenylimidazol-1-yl-oxy)valerate;
8-(1,4,5-Triphenylimidazol-2-yl-oxy)octanamide;
8-[1,4,5-Triphenylimidazol-2-yl-oxy]octanoic acid; or
8-[1,4,5-triphenylimidazol-2-yl-oxy]octanoic acid ammonium salt.

22. The method according to claim 20 or 21 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some part by the lipid inflammatory mediators.

23. A method for treating disease or disorders mediated by the lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprising administering to a mammal in need thereof an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a compound selected from
7-(3,4,5-Triphenylimidazol-1-yl-oxy)heptanitrile;
8-(2,3-Diphenylmaleimido)octanoic acid;
11-(2,3-Diphenylmaleimido)undecanoic acid;
1-(7-Ethoxycarbonyl)-4-phenylimidazole;
Methyl-7-(3,4,5-triphenyl)-2-oxo-1,2-dihydroimidazol-1-yl)-5-heptynoate;
2-[4-(3-Carboxypropoxy)phenyl]-4,5-diphenylimidazole;
1-(7-Carboxyheptyl)-2-phenylimidazole;
1-(7-Ethoxycarbonyl)-4-phenylimidazole;
1-(7-Carboxyheptyl)-2-octylthio-4,5,-diphenylimidazole;
8-(1,4,5-Triphenylimidazol-2-yl-oxy)octanamide; and the pharmaceutically acceptable salts thereof.

24. The method according to claim 23 wherein the compound is
1-(7-Carboxyheptyl)-2-octylthio-4,5,-diphenylimidazole;
8-[1,4,5-Triphenylimidazol-2-yl-oxy]octanoic acid;
Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;
Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate;
Ethyl 6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
7-(3,4,5-Triphenylimidazol-2-oxo-2,3-dihydroimidazol-1-yl)heptanonitrile;
Ethyl 6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
1-(7-Ethoxycarbonyl)-4-phenylimidazole; and
Methyl-7-(3,4,5-triphenyl)-2-oxo-1,2-dihydroimidazol-1-yl)-5-heptynoate.

25. The method according to claims 23 or 34 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome mediated in some part by the lipid inflammatory mediators.

26. The compound which is
1-(7-Carboxyheptyl)-2-octylthio-4,5,-diphenylimidazole;
8-(2,3-Diphenylmaleimido)octanoic acid;
11-(2,3-Diphenylmaleimido)undecanoic acid;
1-(7-Ethoxycarbonyl)-4-phenylimidazole;
7-(3,4,5-Triphenylimidazol-2-oxo-2,3-dihydroimidazol-1-yl)heptanonitrile;
Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate;
Ethyl 6-(3,4,5-triphenyl1.2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;
9-[1-(3,4,5-Triphenyl-2-oxo-2,3-dihydroimidazolyl)]nonanoic acid;
Ethyl 6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
Ethyl-8-(4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanoate;
7-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)heptanitrile; or
Methyl-7-(3,4,5-triphenyl)-2-oxo-1,2-dihydroimidazol-1-yl)-5-heptynoate.

27. The compound according to claim 26 which is
1-(7-Carboxyheptyl)-2-octylthio-4,5-diphenylimidazole; or
Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate.

28. A pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a compound selected from the group consisting of:
1-(7-Carboxyheptyl)-2-octylthio-4,5,-diphenylimidazole;
8-(2,3-Diphenylmaleimido)octanoic acid;
11-(2,3-Diphenylmaleimido)undecanoic acid;
1-(7-Ethoxycarbonyl)-4-phenylimidazole;
7-(3,4,5-Triphenylimidazol-2-oxo-2,3-dihydroimidazol-1-yl)heptanonitrile;
Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate;
Ethyl 6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;
9-[1-(3,4,5-Triphenyl-2-oxo-2,3-dihydroimidazolyl)]nonanoic acid;
Ethyl 6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;
Ethyl-8-(4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanoate;
7-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)heptanitrile; or
Methyl-7-(3,4,5-triphenyl)-2-oxo-1,2-dihydroimidazol-1-yl)-5-heptynoate.

29. The compound according to claim 28 which is
1-(7-Carboxyheptyl)-2-octylthio-4,5-diphenylimidazole; or
Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate.

30. The method according to claim 2 wherein the disease or disorder is allergic rhinitis, asthma, myocardial infarction, stroke, circulatory shock, hypotension, ischemia, reperfusion injury, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, adult respiratory distress syndrome, analphylaxis, shock, endotoxic shock, actinic keratosis, psoriasis, contact dermatitis, pyresis, or any other disease, disorder or syndrome in which a significant increase in the amount, or production of, a lipid inflammatory mediator, arachidonic acid, its metabolites, and/or platelet activating factor occurs.
Description
FIELD OF THE INVENTION

The invention relates to the area of inflammatory mediators. The invention is based on the discovery that blocking a key enzyme responsible for arachidonate movement (or remodelling), Coenzyme A-independent transacylase (CoA-IT), inhibits the production of lipid mediators (free arachidonic acid, arachidonic acid metabolites, and platelet-activating factor (PAF)). It has been discovered that CoA-IT is required to maintain arachidonate content in cellular phospholipids, for the release of free arachidonic acid and the synthesis of arachidonic acid metabolites and PAF. As CoA-IT is involved in arachidonate phospholipid metabolism, and required for the release of free arachidonaic acid and the production of eicosanoids and PAF, inhibition of such would be useful for the treatment of disease states caused thereby.

BACKGROUND OF THE INVENTION

An early event in the response of most inflammatory cells to immunologic activation and other stimuli is the release of newly formed products (mediators) which alter the function and biochemistry of surrounding cells and tissues. The ensuing biological responses, as well as much of the pathogenesis which is attributed to inflammation and allergy, are thought to be dependent on the effects that these newly-formed mediators have on adjacent cells within the inflammatory region.

In the last 20 years, it has become apparent that lipid mediators are among the most potent and important products which are generated during inflammatory reactions. The synthesis of most lipid mediators is initiated by the cleavage of complex phospholipid molecules which contain arachidonate at their sn-2 position. Free arachidonic acid is released from these phospholipids and this represents the rate-limiting step in the formation of eicosanoids (leukotrienes, prostaglandins and thromboxanes). As arachidonic acid is released, it is then converted to oxygenated derivatives by at least two enzymatic systems (lipoxygenase and/or cyclooxygenase). Concomitant with arachidonate release, lysophospholipids are formed. One of these lyso phospholipids, 1-alkyl-2-lyso-sn-glycero-3-phosphocholine, is then acetylated to form platelet-activating factor (PAF). Each of the cell types involved in the inflammatory response produce and secrete a unique subset of lipid mediators. The quantities and nature of the metabolites depend on which enzymes and precursor phospholipid pools are available to inflammatory cells.

Once lipid mediators such as PAF and eicosanoids are formed by the aforementioned pathways, they induce signs and symptoms observed in the pathogenesis of various inflammatory disorders. Indeed, the pathophysiological activity of arachidonic acid (and its metabolites) is well known to those skilled in the art. For example, these mediators have been implicated as having an important role in allergy, asthma, anaphylaxis, adult respiratory distress syndrome, reperfusion injury, inflammatory bowel disease, rheumatoid arthritis, endotoxic shock, and cardiovascular disease. Aalmon and Higgs [Br. Med. Bull (1978) 43:285-296]; Piper et al. [Ann. NY Acad. Sci. (1991) 629:112-119]; Holtzman [Am. Rev. Respir. Dis. (1991) 143:188-203]. Snyder (Am. J. Physiol. Cell Physiol.) (1990) 259:C697-C708]; Prescott et al. [J. Biol. Chem. (1990) 265:17381-17384].

Similar to arachidonate products, PAF is a potent proinflammatory mediator produced by a variety of cells. In vitro, PAF stimulates the movement and aggregation of neutrophils and the release therefrom of tissue-damaging enzymes and oxygen radicals. PAF has also been implicated in activation of leukocytes, monocytes, and macrophages. These activities contribute to the actions of PAF as having (pathological) physiological activity in inflammatory and allergic responses. PAF has also been implicated in smooth muscle contraction, pain, edema, hypotensive action, increases in vascular permeability, cardiovascular disorders, asthma, lung edema, endotoxin shock, and adult respiratory distress syndrome. PAF elicits these responses either directly through its own cellular receptor(s) or indirectly by inducing the synthesis of other mediators.

Accordingly, a method which antagonises the production of free arachidonic acid, its metabolites or PAF will have clinical utility in the treatment of a variety of allergic, inflammatory and hypersecretory conditions such as asthma, arthritis, rhinitis, bronchitis and urticaria, as well as reperfusion injury and other disease involving lipid mediators of inflammation.

Many published patent applications or issued U.S. patents exist which describe various compounds having utility as PAF or Eicosanoid antagonists. Such patents include U.S. Pat. Nos. 4,788,205, 4,801,598, 4,981,860, 4,992,455, 4,983,592,
5,011,847, 5,019,581 and 5,002,941.

Described in this application is a method to inhibit the generation of lipid mediators. As mentioned above, arachidonate-containing phospholipids are the key precursors for a broad range of lipid mediators including arachidonic acid, eicosanoids and PAF. Because of the special role arachidonate-containing phospholipids have in mediator generation, inflammatory cells treat these phospholipids differently than other fatty acid-containing phospholipids. In particular, there are enzymes which control the amount of arachidonate in different phospholipid pools and these enzymes are tightly regulated to maintain arachidonate homeostasis. The movement of arachidonate into and from all phospholipids was originally thought to be exclusively by CoA-dependent acyl transferase activities. Holub et al., Adv. Lipid Res., 16:1-125 (1978); Lands et al., In The Enzymes of Biological Membranes, ed. Martonosi, A., pp. 3-85, Plenum Press, NY, 1976. However, it has now been demonstrated that an enzyme, CoA-IT, is involved in the movment of arachidonate into particular (1-alkyl- and 1-alkenyl) phospholipid pools. These are the phospholipid pools of arachidonate that are preferentially mobilized during cell activation. Moreover, arachidonic acid and lyso-PAF released from these pools are utilized for eicosanoid and PAF, respectively.

CoA-IT has a specificity for certain phospholipids as donor and acceptor molecules. The fatty acid transferred is long chained and unsaturated, and almost exclusively arachidonate. Other fatty acids such as the 16:0, 18:1 or 18:2 are not apparently moved into alkyl and 1-alkenyl phospholipid pools by CoA-IT. The specificity of CoA-IT is in direct contrast to many other CoA-dependent acylation activities which acylate a wide variety of lysophospholipids with no selectivity for arachidonate.

Accordingly, a method by which CoA-IT is inhibited will consequently and preferentially decrease the arachidonate content of 1-alkyl- and 1-alkenyl-linked phospholipids and will therefore decrease the production of pro-inflammatory mediators such as free arachidonic acid, leukotriene and PAF during an inflammatory response. Accordingly, a method by which CoA-IT is inhibited, will have clinical utility in the treatment of allergic, inflammatory and hypersecretory conditions.

SUMMARY OF THE INVENTION

It is an object of this invention to provide a method of treating or reducing allergy and inflammation. It is also an object of this invention to inhibit undesirable lipid mediator production.

This invention is based on the discovery that blocking CoA- independent transacylase, using selective pharmacologic tools, prevents the movement of arachidonate into phospholipid pools needed for the concomitant formation of PAF, free arachidonic acid and its metabolites such as eicosanoids.

The invention relates to a method of treating disease or disorders mediated by free arachidonic acid, its metabolites and/or PAF by administering to a patient in need thereof, an effective amount of a compound which inhibits the production, activation or action of CoA-IT. Inhibition of CoA-IT inhibits lipid mediator production as well as signs and symptoms of disease and disorders induced by lipid mediators.

The premise of this invention is that blocking the movement of arachidonate into specific arachidonate-containing phospholipid pools inhibits lipid mediator (PAF and eicosanoid) production by inflammatory cells. More precisely, when arachidonate is prevented from entering key common precursor phospholipids, precursor molecules will not be formed. If key precursor pools are not formed, arachidonate cannot be removed from these precursors. This means that free arachidonic acid and lyso PAF will be not be mobilized and therefore PAF as well as eicosanoids will not be produced. The end result of CoA-IT inhibition will be reduced signs and symptoms of allergy and inflammation mediated by eicosanoids and PAF.

Still another aspect of the invention relates to a method of screening chemical compounds for potential anti-inflammatory action. In this way, chemical compounds can be rapidly and easily screened for the ability to inhibit CoA-IT and be useful as an anti-inflammatory agent.

Another aspect of the invention relates to the therapeutic use, in medicine,of the compounds, and pharmaceutical compositions, as disclosed herein, in particular for compounds of Formulas (I) to (VI), as inhibitors of CoA-IT activity. As CoA-IT activity is required for the release of lipid inflammatory mediators, such as arachidonic acid and the production of platelet-activiating factor, by inflammatory cells and that inhibition of the production, activation or activity of CoA-IT will have beneficial and therapeutic effect the compounds of the present invention, as described herein, which are inhibitors of CoA-IT are useful in the treatment of disease states caused thereby.

Treatment of disease states caused by these lipid inflammatory mediators i.e., arachidonate, eicosanoids and PAF, include certain cardiovascular disorders such as but not limited to, myocardial infarction, stroke, circulatory shock, or hypotension, ischemia, reperfusion injury, inflammatory diseases such as, but not limited to, arthritis, inflammatory bowel disease, Crohn's disease, or ulcerative colitis, respiratory disease such as but not limited to, asthma, or adult respiratory distress syndrome, analphylaxis, shock such as but not limited to endotoxic shock, topical disesases, such as but not limited to actinic keratosis, psoriasis, or contact dermatitis, or pyresis.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings contain the following figures:

FIG. 1 illustrates the Role of CoA-Independent Transacylase in Arachidonic Acid and Platelet-Activating Factor Metabolism.

FIG. 2 illustrates the effects of the compound of Example 3, Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-1-yl)heptane-phosphonate on CoA-IT activity.

FIG. 3 illustrates the effects of the compound of Example 3, Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-1-yl)heptane-phosphonate which produce a concentration-dependent blockade in the movement of [.sup.3 H]arachidonate into
1-ether-linked phospholipids, 1-alkyl PC and 1-alkenyl PE.

FIG. 4 illustrates the effects of the compound of Example 3, Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-1-yl)heptane-phosphonate which induced a concentration-dependent loss of arachidonate from all major phospholipid classes of the mast cell. In contrast, there was no loss in the mass of 18:0, 18:1, 18:2 from PI, PE and PC in response to Compound 3.

FIG. 5 illustrates the effects of the compound of Example 3, Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-1-yl)heptane-phosphonate on inhibition of the production of PAF in human neutrophils.

FIG. 6 illustrates the effects of the compound of Example 3, Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-1-yl)heptane-phosphonate on inhibition of free arachidonic acid release in a concentration dependent fashion.

FIG. 7 illustrates the in vivo anti-inflammatory responses of the compound of Example 3, Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-1-yl)heptane-phosphonate in the mouse ear by the topical application of a pro-inflammatory agent,
12-0-tetradecanoylphorbol 13-acetate.

DETAILED DESCRIPTION OF THE INVENTION

It has now been discovered that CoA-IT activity is required for lipid mediator production. Specifically, it has been discovered that CoA-IT activity is required for the movement of arachidonate into, and the maintance of arachidonate within, phospholipid pools from which it can be released to form free arachidonic acid and for the production of lyso PAF needed for PAF synthesis. Further, CoA-IT has been shown to be crucial in the mobilization of lyso-PAF and free arachidonic acid during inflammatory cell activation. Inhibition of CoA-IT activity will result in a decrease in arachidonate content of phospholipids, a decreased production of PAF, and a decreased release of free arachidonic acid from cellular phospholipids.

More specficially, FIG. 1 shows a simplified scheme of how arachidonic acid is directed through phospholipids of inflammatory cells. As arachidonic acid enters inflammatory cells or is produced within these cells, it is converted to arachidonoyl-CoA by the enzyme arachidonoyl CoA synthetase. At that point, arachidonic acid is incorporated into the sn-2 position of a lyso phospholipid by arachidonoyl-CoA acyl transferase. The arachidonate-containing phospholipids formed in this reaction appear to belong to a special group or pool (pool 1) of phospholipids which contains predominantly 1-acyl-linkages at the sn-1 position of the molecule. When cells are not stimulated, arachidonic acid is slowly transferred from this first pool to other pools (pool 2) of phospholipids which contain predominantly 1-alkyl and 1-alk-1-enyl linkages at the sn-1 position and phosphatidylcholine and phosphatidylethanolamine linkages at the sn-3 position of phospholipids. This transfer into other pools of AA-containing phospholipids is accomplished by the enzyme CoA-IT.

During inflammatory cell stimulation, there is a calcium-dependent activation of an enzyme phospholipase A.sub.2 which removes arachidonic acid from arachidonate-containing phospholipids which are predominantly in the second (1-alkyl and
1-alk-1-enyl) pool (pool 2). Arachidonic acid and lyso phospholipids formed in this reaction become key intermediates for eicosanoid generation and platelet activating factor generation, respectively. In particular, one of these arachidonate-containing phospholipids in pool 2, 1-alkyl-2-arachidonyl-sn-glycero-3-phosphocholine, is a common precursor for arachidonate and platelet-activating factor. During inflammatory cell activation, arachidonic acid is rapidly depleted from phospholipids in pool 2. As these pools are depleted by the action of phospholipase A.sub.2, they are rapidly replenished by CoA-IT. It is our discovery that the movement of arachidonate into special pools mediated by CoA-IT is required for lipid mediator production and that the blockage of CoA-IT will decrease arachidonate content within cellular phospholipids and inhibit lipid mediator production. This will have beneficial therapeutic effects for diseases mediated, in some part, by eicosanoids and platelet-activating factor.

1. Characteristics of CoA-IT Activity

CoA-IT activity had been defined to have the following characteristics.

A. Co-factors

CoA-IT activity is independent of the presence of Coenzyme A. In addition, no other co-factors required for activity or that modulate activity have been discovered. CoA-IT activity is not altered by the absence or presence of calcium (0-10 mM), magnesium (0-10 mM), EGTA (0-2 mM), EDTA (0-10 mM), ATP, CoA or CoA-fatty acids.

B. pH

CoA-IT activity over a wide range of pH levels was determined. The results demonstrate that the enzyme is active over a broad pH range of 6.5-9. The activity of the enzyme rapidly decreases below pH 6.5 and above pH 10.

C. Kinetics.

The kinetics of the CoA-IT reaction were studied with various concentrations of 1-alkyl-2-lyso-GPC. CoA-IT activity increases as a function of the concentration of substrate, 1-alkyl-2-lyso-GPC. The enzyme exhibits an apparent substrate affinity (K.sub.m) for 1-alkyl-2-lyso-GPC of 0.1-2 .mu.M.

D. Other Characteristics

CoA-IT is stable when treated with dithiothreitol (DTT) or 2-mercaptoethanol (1-10 mM). CoA-IT is inactivated by exposure to heat or acid and is inhibited by addition of detergents such as 3-octyl glucoside, deoxycholate, cholate, Triton X-100, C12E8, CHAPS and hexadecyl-trimethyl ammonium bromide.

E. Specificity

A key characteristic of CoA-IT is the exquisite specificity of this enzyme for polyunsaturated fatty acids and especially arachidonic acid. Sugiura et al. (J. Biol. Chem. (1987) 262:1199-1205); Chilton et al. (J Biol. Chem. (1983)
258:7268-7271); Kramer and Deykin (J. Biol. Chem. (183) 258:13806-13811).

F. Location

Within the cell, CoA-IT activity is completely and tightly associated with microsomal membranes. Treatment of these membranes with 2M KCI fails to extract more than 75% of the CoA-IT activity, suggesting that CoA-IT is an integral membrane component. The subcellular location of CoA-IT activity remains to be determined.

Evidence of CoA-IT activity exists in a variety of inflammatory cells, including human neutrophils, monocytes, lung mast cells, guinea pig eosinophils and human U937 monocytic and HL-60 granulocyte cells lines. There is also preliminary evidence that somewhat less CoA-IT activity is found in tissues such as lung, liver and kidney. Less activity yet is found in heart, skeletal muscle and brain.

G. Comparison with other enzymes

CoA-IT has characteristics which distinguish its activity from the activities of other enzymes involved in lipid metabolism, such as phospholipase A.sub.2, lypoxygenases, cyclooxygenases, CoA-dependent acyltransferases and PAF acetyl transferase.

These differences include different co-factor requirements, location within cells, effects of detergents on activity, effects of heat or acid treatment, stability to reducing agents such as dithiothreitol (DTT) and selectivity for arachidonate-containing substrates. The following Table, Table I, summarizes these differences in characteristics between CoA-IT and other enzymes.

TABLE I ______________________________________ Comparison of CoA-IT to other enzymes ______________________________________ LMW HMW Property CoA-IT Pan.PLA.sub.2 PLA.sub.2 PLA.sub.2 ______________________________________ Co-factors None Ca.sup.2+ Ca.sup.2+ Ca.sup.2+ Location membrane extracellular extra- cytosol cellular Detergent inhibition stimulation inhibition stimulation Heat/Acid unstable stable stable unstable DTT no effect inhibition inhibition no effect AA sel yes no no yes ______________________________________ Property CoA-IT CoA-D AcetylTase AcetylHy ______________________________________ Co-factors None CoA Ca.sup.2+ / none A-CoA Location membrane membrane membrane cyto/LDL Detergent inhibition mixed no effect mixed Heat/Acid unstable -- -- stable DTT no effect no effect no effect inhibition AA Sel yes no no no ______________________________________ Property CoA-IT CO 5LO ______________________________________ Co-factors None peroxide peroxide Location membrane membrane cyto-memb Detergent inhibition no effect -- Heat/Acid unstable unstable unstable DTT no effect -- no effect AA Sel yes yes yes ______________________________________ Key to Table 1: Pan. Pancreatic LDL low density lipoprotein CoAD CoAdependant acyltransferase AcetylTase AcetylCoA transferase --no data available LMW low molecular weight HMW high molecular weight cyto cytosol Ca.sup.2+ calcium CO cyclooxygenase 5LO 5lipoxygenase DTT dithiotheritol ACoA AceticCoA AA Sel Arachidonic acid selectivity

This distinction of CoA-IT from the other enzymes based on characteristics is important for several reasons. First, the data indicate that CoA-IT activity is a novel enzyme activity. Second, even though a microsomal preparation is used to assess CoA-IT activity, the distinct characteristics of CoA-IT assure that the assays measure only CoA-IT activity. Finally, the characteristics of CoA-IT demonstrate that the pharmacological utility of inhibition of CoA-IT is unique.

2. CoA-IT inhibition

(a) Evaluation of CoA-IT inhibition in broken cell assays

Inhibitors of CoA-IT activity have now been discovered and characterized. Suitable inhibitors can readily be identified employing the broken cell assay (a) described below. For example, FIG. 2 shows the effect of compound 3 on CoA-IT activity. Often, inhibitors will include an imidazole structure.

(b) Evaluation of CoA-IT inhibitors in intact cells

CoA-IT activity can also be measured in intact cells by following the movement of a pulse of [.sup.3 H]arachidonate as it moves into the "pool 2" phospholipids (1-alkyl and 1-alkenyl phospholipids) in inflammatory cells. These specific studies were performed in intact human neutrophils, which contain CoA-IT activity and transfer arachidonate into 1-alkyl and 1-alkenyl phospholipids. To determine the effects of Compound 3 on this transfer of arachidonate, neutrophils were exposed briefly to [.sup.3 H]arachidonic acid to allow incorporation of the label into 1-acyl phospholipids, treated with various concentrations of Compound 3 and the movement of [.sup.3 H]arachidonate between phospholipid subclasses assessed over the subsequent 2 hours in the absence of cellular stimulation. FIG. 3 shows that Compound 3 produced a concentration-dependent blockade in the movement of [.sup.3 H]arachidonate into 1-ether-linked phospholipids, 1-alkyl PC and 1-alkenyl PE, and at concentrations similar to those needed to block CoA-IT activity in a broken cell preparation. It is important to note that Compound 3 could completely block arachidonate movement, suggesting that not only can CoA-IT mediate this movement, but that CoA-IT activity is required for arachidonate transfer and that no other enzyme can substitute for CoA-IT.

As treatment of intact cells blocks the movement of arachidonate into specific cellular phospholipids, it followed that prolonged treatment of inflammatory cells with CoA-IT inhibitors could deplete their large endogenous pools of arachidonate within phospholipids. To determine that CoA-IT inhibition can alter the amount of arachidonate within cellular phospholipids, mouse bone marrow-derived mast cells were treated with Compound 3 for 24 hours and the mass of arachidonate within total cellular phospholipids was determined. Compound 3 induced a concentration-dependent reduction in the total cellular arachidonate within mast cells, with greater than 75% reduction apparent at 20 .mu.M. As CoA-IT is specific for arachidonate and not other fatty acyl groups, it is critical to compare the effect CoA-IT inhibitors have on arachidonate with other polyunsaturated fatty acids. FIG. 4 illustrates that Compound 3 induced a concentration-dependent loss of arachidonate from all major phospholipid classes of the mast cell. In contrast, there was no loss in the mass of 18:0, 18:1, 18:2 from PI, PE and PC in response to Compound 3. These results show that CoA-IT inhibitors selectively deplete arachidonate, but not other polyunsaturated fatty acids, from inflammatory cells. The magnitude and rapidness of the loss of arachidonate is striking, and suggests a novel mechanism of action and unique therapeutic potential for CoA-IT inhibitors.

3. Role of CoA-IT in PAF Production and AA Release

The molecule 1-alkyl-2-arachidonoyl-GPC has been shown to be a necessary precursor for PAF production (Chilton et al., J. Biol. Chem. (1984) 259, 12014-12020). CoA-IT activity plays two pivotal roles in PAF production, centering on this molecule. First, CoA-IT activity is required for the specific movement of arachidonate into 1-alkyl-2-arachidonoyl-GPC to produce the necessary precursor molecule for PAF. Second, CoA-IT activity has been shown to promote the breakdown of the precursor of PAF, 1-alkyl-2-arachidonoyl-GPC into lyso PAF, to allow PAF production. This CoA-IT mediated production to lyso PAF can be differentiated from PLA.sub.2 activity. CoA-IT activity plays a central and necessary role in the production of PAF.

There is strong evidence that, in activated inflammatory cells, arachidonate is released from specific phospholipid pools. For example, in neutrophils and mast cells the primary source of free arachidonic acid is 1-alkenyl-2-arachidonoyl-GPE. As shown in FIG. 1, CoA-IT activity, due to its unique properties, can replenish this pool with arachidonate to allow and maintain the release of free arachidonic acid. It has now been discovered that CoA-IT activity is necessary and essential for the release of free arachidonic acid and the subsequent formation of bioactive lipid mediators.

To further demonstrate the utility of inhibiting CoA-IT, the compound of Example 3 was shown to inhibit the production of PAF (assay c) and the release of free arachidonic acid (assay b) from human neutrophils. The method of synthesis of this compound and its structural formula is set forth below. This compound inhibited PAF production (FIG. 5) and free arachidonic acid release (FIG. 6) completely and in a concentration dependent fashion. The specificity for inhibition of CoA-IT activity for these compounds and not the activity of other enzymes, such as PLA2 and PAF acetyl transferase, has been demonstrated. These data demonstrate that inhibition of CoA-IT can and will inhibit the production of PAF and the release of free arachidonic acid.

4. Role of CoA-IT in Inflammatory Responses in vivo

The ability of compounds that inhibit CoA-IT to affect in vivo inflammatory responses was assessed. Inflammatory responses were induced in the mouse ear by the topical application of a pro-inflammatory agent, such as 12-0-tetradecanoylphorbol
13-acetate. This produced an edematous response, as measured by increases in ear thickness, as well as increased inflammatory cellular infiltrate, as measured by increases in myeloperoxidase activity as described in the methods. Application of compounds that inhibit CoA-IT had an anti-inflammatory effect, as demonstrated for compound 3 in FIG. 7. This proven anti-inflammatory effect is predictive of therapeutic usefulness in a wide variety of inflammatory diseases and conditions.

5. Assays

(a) Assay for CoA-IT Activity

The following is a method to measure CoA-IT activity and the effects of compounds on CoA-IT activity. The assay is based upon mixing cellular material containing CoA-IT activity with a stable lyso phospholipid such as 1-alkyl-2-acyl-GPC and measuring the production of phospholipid product such as 1-alkyl-2-acyl-GPC occurring in the absence of added CoA or CoA-fatty acids.

Cell Preparation

Any inflammatory cell that contains high levels of CoA-IT activity can be used, such as neutrophils, macrophages or cell lines such as U937 cells. U937 cells were obtained from American Type Culture Collection and grown in RPMI-1640 media (Gibco, Grand Island, N.Y.) supplemented with 10% fetal bovine serum (Hyclone, Logan, Utah) at 37.degree. C., 5%CO.sub.2. Cells were grown without differentiation (basal state) by any agent, such as dimethyl sulfoxide. As used herein, "inflammatory cells" include, but are not limited to neutrophils, macrophages, monocytes, lymphocytes, eosinophils, basophils, and mast cells.

Microsomal preparation

Microsomes were prepared using standard techniques. In this case, cells were washed with a buffer of 250 mM sucrose, 10 mM Tris, 1 mM EGTA, 1 mM MgCl.sub.2, pH 7.4 and ruptured by N.sub.2 cavitation (750 psi, 10 minutes). The ruptured cells were centrifuged 1000.times. g, 5 minutes. The resulting supernatant was centrifuged at 20,000.times.g,.about.20 minutes. Microsomes were prepared from this supernatant by centrifugation at 100,000.times. g, 60 minutes. The resulting pellet was washed once with assay buffer (150 mM NaCl, 10 mM Na.sub.2 KPO.sub.4, 1 mM EGTA, pH 7.4), recentrifuged and the pellet resuspended in assay buffer (4-20 mg protein/ml) and was stored at -80.degree. C. until assayed.

CoA-IT activity

CoA-IT activity was measured in 1.5 ml centrifuge tubes in a total volume of 100 ul. Microsomes were diluted in assay buffer to the desired protein concentration (6-20 ug/tube). The reaction was initiated by addition of [.sup.3 H ]1-alkyl-2-lyso-sn-glycero-3-phosphocholine (GPC) (.about.0.1 uCi/tube) and 1 .mu.M final cold 1-alkyl-2-lyso-GPC in assay buffer with 0.25 mg/ml fatty acid-poor bovine serumalbumin (BSA) (Calbiochem, La Jolla, Calif.). [.sup.3 H]1-alkyl-2-lyso-GPC, approximately 50 Ci/mmol, was from NEN-Dupont (Boston, Mass.) and cold 1-alkyl-2-lyso-GPC was from Biomol (Plymouth Meeting, Pa.). Microsomes were pretreated with desired agents for the desired time (10 minutes) before the addition of [.sup.3
H]1-alkyl-2-lyso-GPC. The reaction was run for the desired time (10 minutes) at 37.degree. C. The reaction was stopped and the lipids extracted by addition of 100 ul of chloroform:methanol (1:2, v/v) followed by 100 ul of chloroform and 100 ul of 1M KCI. The samples were vortexed and centrifuged at high speed in a microfuge for 2-3 minutes. An aliquot of the chloroform-extracted materials were separated, usually by TLC in chloroform/methanol/acetic acid/water (50:25:8:4, v/v), visualized by radioscanning (Bioscan) and the product, [.sup.3 H ]1-alkyl-2-acyl-GPC, was scraped and quantified by liquid scintillation spectroscopy. With this TLC system, the synthetic standards of 1-alkyl-2-lyso-GPC and 1-alkyl-2-acyl-GPC were well separated, with Rf values of approximately 0.25 and 0.65, respectively. Other methods can be used to separate substrate from product, including but not limited to column chromatography, affinity chromatography and post reaction derivitization.

Protein concentration were assessed using the protein assay reagents from Bio-Rad (Richmond, Calif.).

Results

A variety of compounds have been tested in this assay to determine its selectivity and inability to detect trivial, non-selective inhibitors. Inhibitors of 5-lipoxygenase (5-LO) and cyclooxygenase (CO), such as indomethicin, naproxen,
6-(4'-Fluorophenyl)-5-(4-pyridyl)-2,3-dihydroimidzo-[2,1-b]thiazole and 6-(4'-Fluorophenyl)-5-(4-pyridyl)2,3-dihydroimidzo-[2,1-b]thiazole-dioxide had no effect on CoA-IT activity at concentrations up to 100 .mu.M. The anti-oxidant BHT also has no effect at concentrations up to 100 .mu.M. Compounds which complex with phospholipids and inhibit PLA.sub.2 activity, such as quinacrine and aristolochic acid have no effect on CoA-IT activity at concentrations up to 500 .mu.M. Doxepine, a compound reported to inhibit PAF release did not inhibit CoA-IT at concentrations up to 100 .mu.M. Sodiumdiclofenac, reported to decrease leukotriene production by altering arachidonic acid metabolism, had no effect on CoA-IT activity at concentrations up to 500
.mu.M. These results show that the assay for CoA-IT activity is sensitive and selective.

Representative compounds which inhibit CoA-IT activity in a microsomal CoA-IT assay (assay a) at 50 .mu.M are:

1. Ethyl 6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate

2. Sodium7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimldazol-1-yl)-heptanesulphone

3. Diethyl 7-(3-4-5-triphenyl-2-oxo-2-3-dihydroimidazol-1-yl)heptane phosphonate

4. 8-(1,4,5,-Triphenylimidazol-2-yl-oxy)octanoic acid

5. 8-(2-3-Diphenylmaleimido)octanic acid

6. 11-(2,3-Diphenylmaleimido)undecanoic acid

7. Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate

8. Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate

9. Ethyl 5-(1,4-5-triphenylimidazol-1-yl-oxy)valerate

10. 2-(7-Carboxyheptyl)-4,5-diphenyloxazole

11. Ethyl-6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl-hexanoate

12. Ethyl-8-(4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanoate

13. 8-[1-(1,4,5-Triphenylimidazol-2-yl-oxy)]octanoic acid, ammonium salt

14. 1-(7-Methoxycarbonylheptyl)-4,5-diphenyl-1,2,3-triazole

15. 8-(1,4,5-Triphenyllmidazol-2-yl-oxy)-octanamide

16. 1-(7-Carboxyheptyl)-2-3-4-triphenylimidazole

17. 8-(4,5-Diphenylimldazol-2-yl-thio)octanoic acid

18. 9-[1-(3,4,5-Triphenyl-2-oxo-2,3-dihydroimidazolyl)]nonanoic acid

19. 2-(9-Hydroxynonyl)-4,5-diphenyl-1,2,3-triazole

20. Diethyl 7-(1,4,5-triphenylimidazol-2-yl-oxy)heptane phosphanate

21. -(6-Ethoxycarbonylhexyl)-2,4,5-triphenylimidazole

22. Ethyl 8-(4,5-Diphenylimidazol-1-yl)octanoate

23. 11-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)undecanoic acid

24. 7-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)heptanitrile

25. 7-(3,4,5-Triphenylimidazol-1-yl-oxy)heptanitrile.

26. 1-(6-Carboxyhexyl)-2,4,5-triphenylimidazole

27. 2-(6-Carboxyheptyl)-4-5-diphenyl-1,2,3-triazole

28. 1-(8-Bromooctyl)-4,5-diphenyl-1,2,3-triazole

29. 1-(8-Carboxyoctyl)-2,4,5-triphenylimidazole

30. Ethyl [7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)methyl phosphonate

31. 2-(2-Methoxyethoxy)ethyl-8-(4,5-diphenylimidazol-1-yl)octanoate

32. 1-(8-Cyanooctyl)-4,5-diphenyl-1,2,3-triazole

33. 1-(7 Carboxyheptyl)-2-(4-methoxyphenyl)-4,5-diphenylimidazole

34. 1-(7-Ethoxycarbonylheptyl)-2-methyl-4,5-diphenylimidazole

35. Methyl 7-(3,4,5-triphenyl-2 oxo-2,3-dihydroimidazol-1-yl)-5-heptynoate

36. 2-Benzyl-1-(7-carboxyheptyl)-4,5-diphenylimidazole

37. Ethyl 8-(phenanthro[9,10-d)imidazol-1-yl)octanoate

38. 1-(7-Carboxyheptyl)-2-(4-hydroxyphenyl)-4,5-diphenylimidazole

39. Ethyl 7-(1,4,5-triphenylimidazol-2-yloxy)heptane methylphosphinate

40. 2-[4-(3-Carboxypropoxy)phenyl]-4,5 diphenylimidazole

41. 1-(7-Carboxyheptyl)-4,5,-bis(2-chlorophenyl)-2-phenylimidazole

42. 1-(7-Carboxyheptyl)-2-(4-hydroxy-3,5-diiodophenyl)-4,5-diphenylimidazole

43. 1-(7-Carboxyheptyl)-2-phenyl-4,5-bis(4-methoxyphenyl)imidazole

44. 1-(10-Carboxydecyl)-2,4,5-triphenylimidazole

45. 1-(7-Carboxyheptyl)-2-phenylimidazole

46. 1-(7-Ethoxycarbonyl)-4-phenylimidazole

47. 8-(3,4-Diphonylpyrazol-1-yl)octanoic acid

48. 1-(8-carboxy-8,8-dimethyloctyl)-2,4,5-triphenylimidazole

49. 1-(7 Carboxyheptyl)-2-octylthio-4,5-diphenylimidazole

50. 4-[4-(2,4,5-Triphenylimidazol-1-yl)butyloxy]benzoic acid

51. 1-(Carboxyheptyl)-2-heptyl-4,5-diphenylimidazole

52. 1-(7-(5-Tetrazolyl)heptyl]-2,4,5-triphenylimidazole

53. Sodium 7-(2,4,5-triphenylimidazole-1-yl)heptane sulphonate

54. 2-[5-(1,3-dioxalan-2-yl)pentylthio]-1-(7-ethoxycarbonylheptyl)-4,5-dipheny limidazole

56. 7-(2,4,5-Triphenylimidazol-1-yl)heptane phosphonic acid

(b) Arachidonic Acid Release Assay

Preparation of human neutrophils

Human neutrophils were obtained in the laboratory using three different methods. One method used leukophoresis packs from normal humans and neutrophils were isolated using the histopaque-1077 technique. The blood was centrifuged at 300.times. g for 10 minutes. The cell pellets were resuspended in PBS composed of 137 mM NaCI, 8.8 mM Na2HPO4, 1.5 mM KH2PO4, 2.7 mM KCI (Dulbecco's Gibco Laboratories, Long Island, N.Y.) and layered over histopaque-1077 (Sigma, St. Louis, Mo.). The pellets were collected after centrifugation (300.times. g for 30 minutes) and washed once in PBS. The cell pellets were exposed briefly to dionized water to Iyse any erythrocytes. The remaining cells were collected by centrifugation, suspended in PBS, counted and identified after cytospinning and staining. The final leukocyte preparation was of greater than 95% purity and viability.

The second method isolated human neutrophils from fresh heparinized normal blood using the Histopaque-1077 technique. The blood was layered over Histopaque-1077 (Sigma, St. Louis Mo.) and centrifuged at 400.times. g for 30 minutes. The cell pellets were resuspended in 35 ml of PBS and 12 ml of 6% Dextran, followed by Dextran sedimentation at room temperature for 45 minutes. The upper layer was coLiected and further centrifugated for 10 minutes at 1000 rpm. The cell pellets were exposed briefly to deionized water to lyse erythrocytes. The remaining cells were collected by centrifugation, suspended in PBS, counted and identified after cytospinning and staining. The final leukocyte preparation was of greater than 95% purity and viability.

The third method isolated human neutrophils from freshly drawn heparinized normal blood using the Percoll technique. The blood was first treated with 6% Dextran at room temperature for a 1 hour sedmination. The upper layers of plasma were collected and centrifuged at 400.times. g for 10 minutes. The cell pellets were resuspended in Percoll 1.070 g/ml supplemented with 5% fetal bovine serumand layered on discontinuous gradients (1.080, 1.085, 1.090, 1.095 g/ml) followed by centrifugation at 400.times. g for 45 minutes. The neutrophils were collected from interfaces of 1;080 and 1.085 and the 1.085 and 1.090 Percoll densities, followed by a centrifugation at 400.times. g for 45 minutes. The neutrophils were suspended in PBS, counted and identified after cytospinning and staining. The final leukocyte preparation was of greater than 95% purity and viability.

There was no difference noted in the response of the neutrophils nor in the effects of test compounds in neutrophils isolated by the three different techniques.

Treatment of human neutrophils

Neutrophils were suspended in PBS with 1 mM Ca.sup.2+ and 1.1 mM Mg.sup.2+ at concentrations of 5 to 20.times.106 cells per ml. Cells were added to test tubes and treated with the desired compounds for 5 to 10 minutes, then challenged with calcium ionophere A23187, 2 .mu.M, or vehicle control, PBS containing 0.25-1 mg/ml BSA. After 5 to 20 minutes, the reactions were terminated by addition of an equal volume of chloroform:methanol (1:2, v/v) to the samples. [.sup.2 H.sub.8 ]Arachidonic acid (50, 100 or 200 ng) was added as an internal standard and the lipids were extracted by addition of equal volumes of chloroform and distilled water. The samples were vortexed and centrifuged at high speed and the chloroform layer removed to a clean tube.

Assay for free arachidonic acid

The chloroform extract for each sample was evaporated to dryness and the material resuspended in hexane. The hexane was passed through a Silica solid phase column (500 mg), washed 2.times. with hexane and a fatty acid enriched fraction eluted with hexane:ethyl ether (1:1, v/v). Solvents were removed from the samples under a stream of nitrogen then the samples were convened to pentafluorobenzyl esters using pentafluorobenzyl bromide and diisopropylethylamine in acetronitrile. Solvents were removed and samples were suspended in hexane. GC/MS analysis was performed on a suitable instrument, such as a Finnigan MAT TSQ 700 GC/MS/MS/DS (San Jose, Calif.) operated as a single stage quadruple system or a Hewlett-Packard 5890 with a 5989A M5
system.

The peaks corresponding to arachidonic acid and [.sup.2 H.sub.8 ]Arachidonic acid were identified and the areas of those peaks compared and the released arachidonic acid calculated as ng of arachidonic acid for each sample.

Protein concentrations were assessed using the protein assay reagents from Bio-Rad (Richmond, Calif.).

(c) Assay for Production of Platelet-Activating Factor (PAF)

Preparation of human neutrophils:

Blood was obtained from normal humans and neutrophils were isolated as described for the arachidonic acid release assay, above. The final leukocyte preparation was of greater than 95% purity and viability.

Treatment of human neutrophils

Neutrophils were suspended in PBS at concentrations of 5 to 20.times.10.sup.6 cells per ml. Cells were added to test tubes and treated with the desired compounds for 5 to 10 minutes, then challenged with calcium ionophore A23187, 2 .mu.M and
20-30 .mu.Ci of [.sup.3 H]acetic acid (NEN-Dupont, Boston, Mass.), or the vehicle of PBS with 0.25-1 mg/ml of the. After 5 to 20 minutes, the reactions were terminated by addition of an equal volume of chloroform:methanol (1:2, v/v) to the samples and the lipids were extracted by addition of equal volumes of chloroform and distilled water. The samples were vortexed and centrifuged at high speed and the chloroform layer removed to a clean tube.

Assay for PAF

The chloroform from each tube was evaporated to dryness and the material suspended in a small volume of chloroform or chloroform:methanol (25-100 .mu.l) and the total material spotted on a Silica TLC plate. The plates were developed in chloroform/methanol/acetic acid/water (50:25:8:4, v/v) visualized by radioscanning (Bioscan) and the product, [.sup.3 H]PAF, was scraped and quantified by liquid scintillation spectroscopy. With this TLC system, the Rf value for a synthetic standard of PAF was approximately 0.33.

(d) Assay (Method) for TPA-induced Inflammation

Animals:

Male Balb/c inbred mice were obtained from Charle River Breeding Laboratories (Kingston, N.Y.). Within a single experiment mice (22-25 g) were age-matched. These in vivo experiments typically involved use of 5-6 animals/group.

TPA-induced Inflammation:

TPA (12-0-tetradecanoylphorbol 13-acetate) (Sigma Chemical Company) in acetone (4 .mu.g/20 .mu.l) was applied to the inner and outer surfaces of the left ear of BALB/c male mice. The thickness of both ears was then measured with a dial micrometer (Mitutoyo, Japan) at both 2 and 4 hours after treatment, and the data expressed as the change in thickness (10-.sup.3 cm) between treated and untreated ears. The application of acetone did not cause an edematous response; therefore, the difference in ear thickness represented the response to the TPA. After measuring the edema, the inflammed left ears were removed and stored at -70.degree. C. until they were assayed for MPO (myeloperoxidase) activity where appropriate.

Assay of Myeloperoxidase (MPO) in Inflamed Ear Tissue:

On the day of the assay, partially thawed ear tissues were minced and then homogenized (10% w/v) with a Tissumizer homogenizer (Tekmar Co.) in 50 mM phosphate buffer (pH 6) containing 0.5% HTAB. The tissue homogenates were taken through three cycles of freeze-thaw, followed by brief sonication (10 sec). The method of Bradley et al. was used with modifications as described. The appearance of a colored product from the MPO-dependent reaction of o-dianisidine (0.167 mg/ml; Sigma) and hydrogen peroxide (0.0005%; Sigma) was measured spectrophotometrically at 460 nm. Supernatant MPO activity was quantified kinetically (change in absorbance measured over 3 min, sampled at 15-sec intervals) using a Beckman DU-7 spectrophotometer and a Kinetics Analysis package (Beckman Instruments, Inc.). One unit of MPO activity is defined as that degrading one micromole of peroxide per minute at 25.degree. C.

Statistics:

Statistical analysis was done using Student's "t" test. The ED.sub.35 and ED.sub.50 are values which caused a 35% and 50% (respectively) inhibition of the inflammatory response and were calculated by regression analysis of the dose response data.

The compound of Example 3 demonstrated a positive inhibition in this animal model demonstrating a clear utility in the treatment of topically administered dieases associated with inflammation as noted herein such as, but not limited to, inflammatory bowel disease, contact dermatoses, actinic keratosis, psoriasis, or conjuctivitis.

Alternatively, a dosage of 50 .mu.M/kg per os dose may be administered to the animals and the assay conducted accordingly. A positive in vivo response would similarly be indicative for use in disease states which require systemic treatments, as described herein, such as, but not limited to, asthma, adult respiratory distress syndrome or allergic responses.

(e) Methods for the evaluation of CoA-IT inhibitors in intact cells

Measurement of the effect of CoA-IT inhibitors on the transfer of [.sup.3 H]arachidonate into 1-ether phospholipids in non-stimulated inflammatory cells can be accomplished by general application of the following specific method. Human neutrophils were isolated and resuspended (5.times.10.sup.7 /ml) in Hanks Balanced Salt Solution (HBSS; Gibco). [5,6,8,9,11,12,14,15-.sup.3 H]-Arachidonic acid (100 Ci/mmol; New England Nuclear) complexed to 200 .mu.l HBSS containing 0.25 mg/ml HSA was added to the cell suspension (1 .mu.Ci/ml). The cells were incubated with gentle shaking at 37.degree. C. for 5 min. The reaction was terminated by the addition of 40 ml ice-cold HBSS containing HSA (0.25 mg/ml). The cells were then removed from the supernatant fluid by centrifugation (225 g, 8 min). Unincorporated [.sup.3 H]-arachidonic acid was completely removed by two more washes of HBSS containing 0.25 mg/ml HSA. The neutrophils were resuspended in fresh buffer, exposed to various concentrations of a CoA-IT inhibitor or its vehicle and incubated without stimulation for 2 hrs. At that time, the tubes containing the cells and buffer were extracted (Bligh & Dyer [Can. J. Biochem. Physiol. (1959) 37, 911-917]) and the phospholipid classes separated and collected by normal phase HPLC, using a Ultrasphere Silica column (4.6 mm.times.250 mm; Rainin) eluted with hexane/2-propanol/ethanol/phosphate buffer (pH 7.4)/acetic acid (490:367:100:30:0.6 v/v) for 5 min at a flow rate of 1
ml/min. The amount of phosphate buffer in the eluting solvent was increased to 5% over 10 min and this solvent composition was maintained until all the phospholipid classes had eluted from the column (30-40 min) (Chilton, F. H. [Methods Enzymol. (1990)
187, 157-166]). The phospholipids were converted into diradylglycerols by addition of phospholipase C, 20 units-40 units of Bacillus cereus phospholipase C (Sigma Type XIII) in 100 mM Tris HCl buffer (pH 7.4) for 2.5-6 hr, then converted into
1,2-diradyl-3-acetylglycerols by incubation with acetic anhydride and pyridine (Chilton, F. H. [Methods Enzymol. (1990) 187, 157-166]). The phospholipid subclasses were separated by TLC in benzene/hexane/ethyl ether (50:45:4, v/v), located by image analysis (Bioscan) and the amount of radioactivity in each class was determined by zonal scraping and liquid scintillation counting.

The following is the method for assessing the ability of a compound to alter arachidonate content of cellular phospholipids, which can be generalized for any desired cell. Specifically, mouse bone marrow-derived mast cells are removed from culture and provided with exogenous [.sup.3 H]arachidonic acid for 30 minutes. The labeled arachidonic acid which had not been incorporated into the cells is then removed by washing the cells 2 times with an albumin-containing buffer. At that point, the cells are treated with various concentrations of CoA-IT inhibitors and then placed back in culture for 24-48 hours. The phospholipids are extracted by the method of Bligh and Dyer [Can. J. Biochem. Physiol. (1959) 37, 911-917] and phospholipids separated by normal phase HPLC by the method of Chilton [Methods Enzymol. (1990) 187, 157-166]. The radioactive and mole quantities of arachidonate in complex lipids are determined. At this point, cellular lipid extracts are treated with KOH (0.5M) to remove fatty acids from complex lipids (phospholipids) and the quantities of arachidonate in these extracts can then be determined by various methods, including gas chromatography and mass spectrometry (Chilton [Methods Enzymol. (1990) 187, 157-166]).

6. Assay for screening chemical compounds for potential anti-inflammatory action

An assay method for determining the inhibitory activity of compounds for CoA-IT and the inhibition of PAF and free arachidonic acid production is also encompassed by the invention. The method comprises (1) measuring the inhibition of the CoA-independent acylation of lysophospholipids in broken cell preparations of said compounds; (2) measuring the inhibition of PAF production in activated inflammatory cells of said compounds; and/or (3) measuring the inhibition of free arachidonic acid release in activated inflammatory cells of said compounds; and/or (4) measuring deletion of arachidonate form phospholipds of inflammatory cells by said compounds; and/or (5) measuring the anti-inflammatory activity of said compounds in animal models of inflammation. The activity of the compound is determined by inhibition of at least 20% of the activities of CoA-IT, PAF or free arachidonic acid release. This assay method provides a means wherein chemical compounds can be easily screened for CoA-IT inhibiting activity.

As used herein, various abbreviations and explanations are as follows: [.sup.3 H], a molecule that contains tritium atoms, a radioactive isotope; A23187, a compound that allows free entry of calcium into a cell; AA, arachidonic acid; arachidonate, arachidonic acid contained within a phospholipid; free arachidonic acid, arachidonic acid that is not contained within a phospholipid; [.sup.2 H.sub.8 ]arachidonic acid, the form of arachidonic acid labeled with 8 deuterium atoms, a stable isotope; 1-alkyl, 1-O-alkyl; 1-alkenyl, 1-O-alk-1'-enyl; BSA, bovine serum albumin; CoA, coenzyme A; CoA-IT, CoA-independent transacylase; DTT, dithiothreitol; EGTA, [ethylenebis(oxyethylenenitrilo)]tetra acetic acid, a calcium chelator; GPC, sn-glycero-3-phosphocholine; EDTA, a metal ion chelator; GPE, sn-glycero-3-phosphoethanolamine; GC/MS, gas chromatography and mass spectrometry; 5HETE, 5(S)-hydroxyeicosa-6,8,11,14-tetraenoic acid; 15HETE, 15(S)-hydroxyeicosa-5,8,11,13-tetraenoic acid; HL-60, American Type Tissue Culture designated cell line similar to a monocyte; LTB.sub.4, leukotriene B.sub.4 ; LTC.sub.4, leukotriene C.sub.4 ; LTD.sub.4, leukotriene D.sub.4 ; lyso PAF, 1-alkyl-2-lyso-GPC, lyso platelet-activating factor; PLA.sub.2, phospholipase A.sub.2 ; PBS, phosphate buffered saline; PAF, platelet activating factor, 1-alkyl-2-acetyl-GPC; PL, phospholipid; PC, phosphatidylcholine; PE, phosphatidylethanolamine, PI, phosphatidylinositol; PMN, polymorphonuclear neutrophilic cell, neutrophil; PS phosphatidylserine; Rf, the distance a compound travels as a fraction of the solvent front; TLC, thin layer chromatography; U937, American Type Tissue Culture designated cell line similar to a monocyte.

Compounds

Illustrative of compounds useful in this inventions are the compounds of Formulas (I) to (VI), as noted below. Compounds which are also useful in the instant invention and which do not specifically fall within any of the structures herein are further described below. Another invention is the pharmaceutical compositions for use herein comprising the compounds as noted herein, and in particular the pharmacuetical compositions comprising a compounds of Formulas (I) to (VI), or a pharmacuetically acceptable salt thereof and a pharmacuetically acceptable carrier or diluent.

Compounds of Formula (I) are represented by the structure: ##STR1## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl;

n is an integer having a value of 4 to 12;

X is 5-tetrazolyl, SO.sub.3 H, P(O)(OR.sub.2).sub.2, P(O)(OH).sub.2, or P(O)(R.sub.2)(OR.sub.2);

R.sub.2 is hydrogen or C.sub.1-4 alkyl;

R.sub.3 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;

m is an integer having a value of 1 to 3;

q is an integer having a value of 1 to 3;

or a pharmaceutically acceptable salt thereof.

Suitably, R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl. Preferably R.sub.1 is optionally substituted phenyl; most preferably an unsubstituted phenyl.

Suitably, n is 4 to 12; preferably n is 4 to 8, most preferably n is 6 or 7.

Suitably m and p are 1 to 3, preferably 1.

Suitably, X is 5-tetrazolyl, SO.sub.3 H, P(O)(OR.sub.2).sub.2, P(O)(OH).sub.2, or P(O)(R.sub.2)(OR.sub.2) in which R.sub.2 is independently a C.sub.1-4 alkyl group. Preferably X is P(O)(OEt).sub.2 or P(O)(Me)(OEt).

Suitable R.sub.3 substituent include, for example, 1 to 3 groups which may be the same or different and are selected from C.sub.1-4 alkyl, such as methyl or ethyl, haloC.sub.1-4 alkyl such as CF.sub.3, halogen, such as F or Cl, hydroxy and C.sub.1-4 alkoxy, such as methoxy. Preferably R.sub.3 is hydrogen.

Suitable heteroaryl groups include, for example, saturated or unsaturated 5- or 6- membered rings comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulphur.

Preferably such rings include, for example, thienyl and furyl rings.

Compounds of structure (I) include:

1. Diethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate;

2. Ethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)methyl-phosphinate

3. 7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonic acid;

4. Sodium 7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane sulphonate;

5. Diisopropyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate;

6. Dimethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate;

7. Diethyl-6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexane phosphonate; or

8. Diethyl-8-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octane phosphonate.

A preferred compound of Formula (I) is 7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)-heptanephosphonate.

Another aspect of the present invention is the novel compounds and their pharmaceutical compositions of Formula (I) which are:

Diisopropyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate;

Dimethyl-7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate;

Diethyl-6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexane phosphonate;

Diethyl-8-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octane phosphonate; and the pharmaceutically acceptable salts thereof.

The compounds of structure (I) can be prepared using procedures analogous to those known in the art. The present invention therefore provides in a further aspect a process for the preparation of compounds of structure (I) in which X is other than 5-tetrazolyl which comprises reaction of a compound of structure (Ia): ##STR2## in which

R.sub.1, R.sub.3, m, n, and p are as described for structure (I) and L is a leaving group, with a suitable source of the group X; and optionally thereafter forming a pharmaceutically acceptable salt thereof.

Compounds of structure (I) in which X is 5-tetrazolyl, can be prepared from compounds of structure (Ia) by standard techniques, for example, when L is bromine, by reaction with sodium cyanide in a suitable solvent such as dimethylsulphoxide, to form the intermediate compound in which L is cyano; followed by reaction with tri-n-butyl tin azide in, for example, tetrahydrofuran to form the desired compound of structure (I).

Suitable leaving groups L will be apparent to those skilled in the art and include, for example, halogen, such as bromine.

Suitable sources of the group X will again be apparent to those skilled in the art and include, for example, where X is SO.sub.3 Na, sodium sulphite.

The reaction between the compounds of structure (Ia) and the source of X is carried out in a solvent at elevated temperature. Preferably, for example where X is SO.sub.3 Na the reaction is carried out in aqueous ethanol at reflux temperature for a suitable period to allow the reaction to go to completion; and where X is a phosphorus containing group the reaction is carried out in an organic solvent such as toluene or xylene.

The compounds of structure (Ia) can be prepared from compounds of structure (Ib): ##STR3## in which R.sub.1, R.sub.3, m, n, and p are as described for structure (I) by reaction with, for example, a compound of formula L.sup.1 (CH.sub.2).sub.n L, in which L and L.sup.1 are suitable leaving groups, in the presence of a base such as potassium carbonate and a suitable solvent such as butanone. Suitable groups L are as described for structure (Ia). Suitable groups L.sup.1 will be apparent to those skilled in the art, and include halogen, in particular bromine.

Compounds of structure (Ib) are known or can be prepared by standard techniques.

The compounds of Examples 1 to 8 found in the Synthetic Chemistry section serve to illustrate the preparation of compounds representative of structure (I).

Compounds of Formula (II) are represented by the structure ##STR4## wherein R is hydrogen, C.sub.1-8 alkyl, C.sub.1-8 alkoxy, SC.sub.1-8 alkyl, optionally substituted phenyl, phenyl C.sub.1-4 alkyl in which the phenyl group is optionally substituted, C.sub.1-6 alkylCHO or C.sub.1-6 alkylCH(OR.sup.1)(OR.sup.2) in which each group R.sup.1 and R.sup.2 is C.sub.1-4 alkyl, or together form an ethane 1,2-diyl or propane 1,3-diyl group;

n is 2 to 6 and m is 0 to 6;

R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are independently hydrogen or C.sub.1-4 alkyl;

AB is a single bond, --CH.dbd.CH--, --S--, S-phenyl or O-phenyl;

X is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl;

R.sup.7 is independently selected from hydrogen, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, halogen, hydroxy, or C.sub.1,4 alkoxy;

p is an integer having a value of 1 to 3;

or a pharmaceutically acceptable salt thereof;

provided that:

a) when X is 5-tetrazolyl, R.sup.7 is hydrogen, R is phenyl, and AB is a bond, then n+m are equal to a number greater than 6;

b) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not hydrogen;

c) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not alkyl or hydrogen;

d) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 4-hydroxy, then R is not phenyl;

e) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 4-Methoxy or is 4-hydroxy, then R is not hydrogen;

f) when X is CO.sub.2 H, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 2-chloro, then R is not hydrogen;

g) when (R.sup.7)p is the same and is hydrogen, R is phenyl, n is 4, m is 0, and AB is O-phenyl then X is not CO.sub.2 --C.sub.1-6 alkyl;

h) when R is hydrogen, (R.sup.7)p is the same and is hydrogen, AB is a bond, n+m is equal to 7, than X is not CH.sub.3 O--(CH.sub.2).sub.2 --O--(CH.sub.2).sub.2 --O--C(O)--;

i) when X is CO.sub.2 --C.sub.1-6 alkyl; AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not phenyl or 4-methoxyphenyl;

j) when X is CO.sub.2 --C.sub.1-6 alkyl, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is 4-bromo or 4-methoxy, then R is not hydrogen;

k) when X is CO.sub.2 --C.sub.1-6 alkyl, AB is a bond, n+m is equal to 7, and (R.sup.7)p is the same and is hydrogen, then R is not 2-(4-methoxybenzyl);

l) when (R.sup.7)p is the same and is hydrogen, R is phenyl, AB is a bond n+m is equal to 10, then X is not CO.sub.2 --C.sub.1-6 alkyl;

m) when (R.sup.7)p is the same and is hydrogen, R is phenyl, n is 4, m is 0 and AB is O-phenyl, then X is not CO.sub.2 --C.sub.1-6 alkyl;

n) when AB is --S--, n is 5 or 6, and m is 1 then X is CO.sub.2 H; or a pharmaceutically acceptable salt thereof.

Suitably, p is 1 to 3, and R.sup.7 is independently selected from hydrogen, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, such as CF.sub.3, halogen, hydroxy or C.sub.1-4 alkoxy. Preferably R.sup.7 is hydrogen.

Suitably, R is hydrogen, C.sub.1-8 alkyl, C.sub.1-8 alkoxy, SC.sub.1-8 alkyl, optionally substituted phenyl, phenyl C.sub.1-4 alkyl in which the phenyl group is optionally substituted, C.sub.1-6 alkylCHO or C.sub.1-6 alkylCH(OR.sup.1)(OR.sup.2) in which each group R.sup.1 and R.sup.2 is C.sub.1-4 alkyl, or together form an ethane 1,2-diyl or propane 1,3-diyl group.

Preferably R is C.sub.1-4 alkyl or optionally substituted phenyl. When R is an optionally substituted phenyl the substituent include, for example, 1 to 3 groups which may be the same or different and are selected from C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, such as CF.sub.3, halogen, hydroxy and C.sub.1-4 alkoxy.

Suitably, n and m together are 4 to 12, preferably 4 to 8, and most preferably 6 or 7.

Suitably, R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are the same or different and are each hydrogen or C.sub.1-4 alky; Preferably, R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are the same and are each hydrogen.

Suitably, AB is a single bond, --CH.dbd.CH--, S-phenyl or O-phenyl. Preferably, AB is a single bond.

Suitably, X is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl. Preferably X is CO.sub.2 H, a group hydrolysable to CO.sub.2 H or 5-tetrazolyl.

Suitable heteroaryl groups include, for example, saturated or unsaturated 5- or 6-membered rings comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulphur. Preferably such rings include, for example, thienyl and furyl rings.

Suitable groups X, hydrolysable to CO.sub.2 H include for example, nitriles, amides and ester groups. Examples of ester groups are C.sub.1-6 alkyl esters and optionally substituted benzyl esters. Particular ester groups include mono-C.sub.1-4
alkoxycarbonyl groups such as ethoxycarbonyl and methoxycarbonyl, and tri-C.sub.1-4 alkoxy carbonyl groups such as methoxyethoxyethoxy carbonyl groups (CH.sub.3 O(CH.sub.2).sub.2 O(CH.sub.2).sub.2 O--C(O)--).

Compounds of Formula (II) include:

1-(7-Ethoxycarbonylheptyl)-2,4,5-triphenylimidazole;

1-(7-Carboxyheptyl)-2,4,5-triphenylimidazole;

1-(7-Methoxycarbonylheptyl)-2,4,5-triphenylimidazole;

1-(6-Ethoxycarbonylhexyl)-2,4,5-triphenylimidazole;

1-(6-Carboxyhexyl)-2,4,5-triphenylimidazole;

1-(8-Carboxyoctyl)-2,4,5-triphenylimidazole;

1-(10-Carboxydecyl)-2,4,5-triphenylimidazole;

1-(7-Ethoxycarbonylheptyl)-2-methyl-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-methyl-4,5-diphenylimidazole;

1-[7-(5-Tetrazolylheptyl]-2,4,5-triphenylimidazole;

2-(2-Methoxyethoxy)ethyl-8-(2,4,5-triphenylimidazol-1-yl)octanoate;

Ethyl 8-(4,5-diphenylimidazol-1-yl)octanoate;

8-(4,5-Diphenyl-imidazol-1-yl)octanoic acid;

2-(2-Methoxyethoxy)ethyl-8-(4,5-diphenylimidazole-1-yl)octanoate;

1-(7-Ethoxycarbonylheptyl)2-(4-methoxyphenyl)-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-(4-methoxyphenyl)-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-(4-hydroxyphenyl)-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-(4-hydroxy-3,5-diiodophenyl)-4,5-diphenylimidazole;

2-Benzyl-1-(7-ethoxycarbonylheptyl)-4,5-diphenylimidazole;

2-Benzyl-1-(7-carboxyheptyl)-4,5-diphenylimidazole;

1-(7-Ethoxycarbonylheptyl)-2-[4-octyloxyphenyl]-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-[4-octyloxyphenyl]-4,5-diphenylimidazole;

1-(7-Ethoxycarbonylheptyl)-2-octylthio-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2 octylthio-4,5-diphenylimidazole;

1-(7-Ethoxycarbonylheptyl)-4,5-bis-4-hydroxyphenyl)-imidazole;

4,5-Bis(2-chlorophenyl)-1-(7-ethoxycarbonyl-heptyl)imidazole;

4,5-Bis(2-chloro-phenyl)-1-(7-ethoxycarbonylheptyl)-2-phenylimidazole;

1-(7-Carboxyheptyl)-4,5-bis-(2-chlorophenyl)-2-phenylimidazole;

1-(7-Ethoxy-carbonylheptyl)-4,5-bis-(4-methoxyphenyl)-2-phenylimidazole;

1-(7-Carboxyheptyl)-4,5-bis(4-methoxy-phenyl)-2-phenylimidazole;

1-(7-Ethoxycarbonylheptyl)-2-heptyl-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-heptyl-4,5-diphenylimidazole;

7-(1,2,4-Triphenylimidazolyl)-hept-5-ynoic acid;

9-(1,2,4-Triphenylimidazolyl)-2,2-dimethylnonanoic acid;

4-[4-(2,4,5-Triphenylimidazolyl)butyloxy]benzoic acid;

7-(2,4,5-Triphenylimidazol-1-yl)heptanesulphonate;

Sodium 7-(2,4,5-Triphenylimidazol-1-yl)heptanesulphonate;

7-(2,4,5-Triphenylimidazol-1-yl)heptanephosphonate;

7-(2,4,5-Triphenylimidazol-1-yl)heptanephosphonic acid;

Ethyl 8-(phenanthro[9.10-d]imidazol-1-yl)octanoate; or

1-(7-Carboxyheptyl)-2-(5-[1,3-dioxalan-2-yl]pentylthio)-4,5-diphenyl imidazole.

Preferred compounds of Formula (II) include:

1-(7-Carboxyheptyl)-2-heptyl-4,5-diphenylimidazole;

1-(7-(5-Tetrazolylheptyl)-2,4,5-triphenylimidazole;

1-(10-Carboxydecyl)-2,4,5-triphenylimidazole;

4-[4-(2,4,5-triphenylimidazolyl)butyloxy]benzoic acid;

9-(1,2,4-tri-phenylimidazolyl)-2,2-dimethylnonanoic acid;

1-(8-Carboxyoctyl)-2,4,5-triphenylimidazole;

1-(7-Carboxy-heptyl)-2-(4-hydroxy-3,5-diiodophenyl)-4,5-diphenylimidazole;

Ethyl 8-(4,5-diphenylimidazol-1-yl)octanoate;

1-(7-Ethoxycarbonyl-heptyl)-2-methyl-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-(4-hydroxyphenyl)-4,5-diphenylimidazole;

1-(7-carboxyheptyl)-2,4,5-triphenylimidazole;

1-(6-ethoxy-carbonylhexyl)-2,4,5-triphenylimidazole;

1-(6-carboxyhexyl)-2,4,5-triphenylimidazole;

2-(2-methoxyethoxy)ethyl 8-(4,5-diphenylimidazole-1-yl)octanoate;

1-(7-carboxyheptyl)-2-(4-methoxyphenyl)-4,5-diphenylimidazole;

2-benzyl-1-(7-carboxyheptyl)-4,5-diphenylimidazole;

1-(7-carboxyheptyl)-4,5-bis(2-chloro-phenyl)-2-phenylimidazole;

1-(7-carboxyheptyl)-4,5-bis(4-methoxy-phenyl)-2-phenylimidazole;

7-(2,4,5-tri-phenylimidazol-1-yl)heptane-sulphonate;

7-(2,4,5-triphenylimidazol-1-yl)heptanephosphonic acid; or

Ethyl 8-(phenanthrimidazol-1-yl)octanoate.

More preferred compounds of Formula (II) are:

1-(7-Carboxyheptyl)-2-heptyl-4,5-diphenylimidazole;

1-(7-(5-Tetrazolylheptyl)-2,4,5-triphenylimidazole;

1-(10-Carboxydecyl)-2,4,5-triphenylimidazole;

4-[4-(2,4,5-triphenylimidazolyl)butyloxy]benzoic acid;

9-(1,2,4-tri-phenylimidazolyl)-2,2-dimethylnonanoic acid;

1-(8-Carboxyoctyl)-2,4,5-triphenylimidazole;

1-(7-Carboxy-heptyl)-2-(4-hydroxy-3,5-diiodophenyl)-4,5-diphenylimidazole;

Ethyl 8-(4,5-diphenylimidazol-1-yl)octanoate;

1-(7-Ethoxycarbonyl-heptyl)-2-methyl-4,5-diphenylimidazole; or

1-(7-Carboxy-heptyl)-2-(4-hydroxyphenyl)-4,5-diphenylimidazole.

Most preferred compounds of Formula (II) are:

1-(7-Carboxyheptyl)-2-heptyl-4,5-diphenylimidazole;

1-(7-(5-Tetrazolylheptyl)-2,4,5-triphenylimidazole; or

1(10-Carboxydecyl)-2,4,5-triphenylimidazole.

The compounds of structure (II) can be prepared using procedures analogous to those known in the art. The present invention therefore provides in a further aspect a process for the preparation of compounds of structure (II) which comprises:

(a) for compounds other than those in which X is 5-tetrazolyl, reaction of a compound of structure (IIa): ##STR5## in which Ar, R, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, AB, n, p, and m are as described for structure (II) and L is a leaving group, with a suitable source of the group X;

(b) reaction of a compound of structure (IIb): ##STR6## in which R and R.sup.7 are as described for structure (II) with a compound of structure (IIc):

in which R.sup.3, R.sup.4, R.sup.5, R.sup.6, AB, n, m and X are as described for structure (II) and L is a leaving group; or

(c) for compounds in which A is other than a bond or --CH.dbd.CH--, reaction of a compound of structure (IId): ##STR7## in which Ar, R, R.sup.3, R.sup.4 R.sup.7, and n are as described for structure (II) and L is a leaving group, with a compound of structure (IIe):

in which A.sup.1 B.sup.1 is --C.tbd.C--, S, O, SPh or OPh, R.sup.5, R.sup.6, m and X are as described for structure (II) and L' is hydrogen or a metal;

(d) for compounds in which X is 5-tetrazolyl reaction of a compound of structure (IIa) in which L is CN, with tri-n-butyl tin azide, and optionally thereafter converting one group X into another group X, and optionally forming a salt.

Suitable leaving groups L will be apparent to those skilled in the art and include, for example, halogen, such as bromine, and sulphonic acid derivatives such as tosylate and mesylate.

Suitable metals include, for example, alkali metals such as sodium or lithium.

Suitable sources of the group X will again be apparent to those skilled in the art and include, for example, where X is SO.sub.3 Na, sodium sulphite.

The reaction between the compounds of structure (IIa) and the source of X is carried out in a solvent at elevated temperature. Preferably, for example where X is SO.sub.3 Na the reaction is carried out in aqueous ethanol at reflux temperature for a suitable period to allow the reaction to go to completion; and where X is a phosphorus containing group the reaction is carried out in an organic solvent such as toluene or xylene.

The reaction between compounds of structure (IIb) and structure (IIc) can be carried out in an organic solvent in the presence of a base, at a temperature of between ambient and the reflux temperature of the solvent used. Suitable solvents include, for example, C.sub.1-4 alkanols such as methanol or ethanol, dimethyl formamide and butanone, and suitable bases include, for example, potassium carbonate, sodium hydroxide and sodium hydride.

The reaction between compounds of structure (IId) and structure (IIe) is carded out in a suitable solvent in the presence of a base at a temperature of between ambient and the reflux temperature of the solvent used.

Suitable solvents and reagents include, for example, potassium carbonate as the base in butanone as solvent, and sodium in methanol as a solvent.

Compounds of structure (II) in which X is 5-tetrazolyl, can be prepared from compounds of structure (IIa) by standard techniques, for example, when L is bromine, by reaction with sodium cyanide in a suitable solvent such as dimethylsulphoxide, to form the intermediate compound in which L is cyano; followed by reaction with tri-n-butyl tin azide in, for example, tetrahydrofuran to form the desired compound of structure (II).

The intermediate compounds of structures (IIa), (IIb), (IIc), (IId) and (IIe) are known or can be prepared by standard techniques.

Examples 9 to 49 found in the synthetic chemistry section serve to illustrate the preparation of compounds representative of structure (II).

The compounds of Formula (III) are represented by the structure ##STR8## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl;

n is 4 to 12;

Y is oxygen or sulfur;

X is 5-tetrazolyl, cyano, SO.sub.3 H, P(O)(OR.sub.2).sub.2, P(O)(OH).sub.2, or P(O)(R.sub.2)(OR.sub.2);

R.sub.2 is hydrogen or C.sub.1-4 alkyl;

R.sub.3 is independently hydrogen, C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;

m is an integer having a value of 1 to 3;

q is an integer having a value of 1 to 3;

provided that when X is cyano, R.sub.1 is an optionally substituted phenyl;

or a pharmaceutically acceptable salt thereof.

Suitably, R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl. Preferably R.sub.1 is optionally substituted phenyl; most preferably an unsubstituted phenyl.

Suitably, n is 4 to 12; preferably n is 4 to 8, most preferably n is 6 or 7.

Suitably Y is oxygen or sulphur; preferably Y is oxygen.

Suitably m and p are 1 to 3, preferably 1.

Suitably, X is 5-tetrazolyl, SO.sub.3 H, P(O)(OR.sub.2).sub.2, P(O)(OH).sub.2, or P(O)(R.sub.2)(OR.sub.2) in which R.sub.2 is independently a C.sub.1-4 alkyl group. Preferably X is P(O) (OEt).sub.2 or P(O)(Me)(OEt).

Suitable R.sub.3 substituent include, for example, 1 to 3 groups which may be the same or different and are selected from C.sub.1-4 alkyl, such as methyl or ethyl, haloC.sub.1-4 alkyl such as CF.sub.3, halogen, such as F or Cl, hydroxy and C.sub.1-4 alkoxy, such as methoxy.

Suitable heteroaryl groups include, for example, saturated or unsaturated 5- or 6-membered rings comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulphur.

Preferably such rings include, for example, thienyl and furyl rings.

Compounds of structure (III) include:

Sodium 6-(1,4,5-triphenylimidazol-2-yloxy)hexanesulphonate;

Sodium 7-(1,4,5-triphenylimidazol-2-yloxy)heptanesulphonate;

7-(1,4,5-Triphenylimidazol-2-yl-oxy)heptanemethylphosphinate;

7-(1,4,5-Triphenylimidazol-2-yl-oxy)heptanephosphonate;

Ethyl-7-(1,4,5-triphenyl-imidazol-2-yl-oxy)heptane methylphosphinate;

Diethyl-7-(1,4,5-triphenyl-imidazol-2-yl-oxy)heptane phosphonate; and

7-(3,4,5-Triphenylimidazol-1-yl-oxy)heptanitrile.

Preferred compounds of Formula (III) include:

Ethyl-7-(1,4,5-triphenyl-imidazol-2-yl-oxy)heptane methylphosphinate; and

Diethyl-7-(1,4,5-triphenyl-imidazol-2-yl-oxy)heptane phosphonate.

The compounds of structure (III) can be prepared using procedures analogous to those known in the art. The present invention therefore provides in a further aspect a process for the preparation of compounds of structure (III) in which X is other than 5-tetrazolyl which comprises reaction of a compound of structure (IIIa): ##STR9## in which R.sub.1, R.sub.3, m, n, and p are as described for structure (III) and L is a leaving group, with a suitable source of the group X; and optionally thereafter forming a pharmaceutically acceptable salt thereof.

Compounds of structure (III) in which X is 5-tetrazolyl, can be prepared from compounds of structure (IIIa) by standard techniques, for example, when L is bromine, by reaction with sodium cyanide in a suitable solvent such as dimethylsulphoxide, to form the intermediate compound in which L is cyano; followed by reaction with tri-n-butyl tin azide in, for example, tetrahydrofuran to form the desired compound of structure (III).

Suitable leaving groups L will be apparent to those skilled in the art and include, for example, halogen, such as bromine.

Suitable sources of the group X will again be apparent to those skilled in the art and include, for example, where X is SO.sub.3 Na, sodium sulphite.

The reaction between the compounds of structure (IIIa) and the source of X is carried out in a solvent at elevated temperature. Preferably, for example where X is SO.sub.3 Na the reaction is carried out in aqueous ethanol at reflux temperature for a suitable period to allow the reaction to go to completion; and where X is a phosphorus containing group the reaction is carried out in an organic solvent such as toluene or xylene.

The compounds of structure (IIIa) can be prepared from compounds of structure (IIIb): ##STR10## in which R.sub.1, R.sub.3, Y, m, n, and p are as described for structure (III) by reaction with, for example, a compound of formula L.sup.1
(CH.sub.2).sub.n L, in which L and L.sup.1 are suitable leaving groups, in the presence of a base such as potassium carbonate and a suitable solvent such as butanone. Suitable groups L are as described for structure (IIIa). Suitable groups L.sup.1 will be apparent to those skilled in the art, and include halogen, in particular bromine.

Compounds of structure (IIIb) are known or can be prepared by standard techniques.

Examples 50 to 55 found in the synthetic chemistry section serve to illustrate the preparation of compounds represented by structure (III).

The compounds of Formula (IV) are represented by the structure ##STR11## wherein X is nitrogen or CR.sup.1 ;

R.sup.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl;

Y is nitrogen, N(CH.sub.2).sub.n A or C(CH.sub.2).sub.n A

Z is nitrogen, oxygen or N(CH.sub.2).sub.n A', and the dotted line indicates the optional presence of a double bond so as to form a fully unsaturated heterocyclic ring;

n is 4 to 12;

A is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, OH, Br, Cyano, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl;

A' is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl;

R.sup.2 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;

m is a number having a value of 1 to 3; provided that

a) X, Y and Z are not all at the same time, nitrogen;

b) when X is CR.sup.1, Y and Z are not both nitrogen;

c) when Y is N(CH.sub.2).sub.n A, Z is nitrogen; and

d) when Z is oxygen, Y is C(CH.sub.2).sub.n A;

e) when Y is N(CH.sub.2).sub.n A, X and Z are nitrogen, (R.sub.2).sub.m is the same and is hydrogen, and n is 6,7, or 8 then X is not --CO.sub.2 --C.sub.1-6 alkyl;

f) when Z is oxygen, Y is C(CH.sub.2).sub.n A, n is 8, and (R.sup.2).sub.m is the same and is hydrogen, then X is not cyano;

g) when Z is N(CH.sub.2).sub.n A', X is nitrogen, Y is nitrogen, (R.sub.2).sub.m is the same and is hydrogen, and n is 7, then X is not CO.sub.2 H;

h) when Y is N(CH.sub.2).sub.n A, X and Z are nitrogen, (R.sub.2).sub.m is the same and is hydrogen, and n is 8 then X is not cyano; or a pharmaceutically acceptable salt thereof.

Suitably, X is nitrogen or CR.sup.1 ; preferably X is nitrogen.

Suitably, Y is nitrogen, N(CH.sub.2).sub.n A or C(CH.sub.2).sub.n A; preferably, Y is nitrogen or N(CH.sub.2).sub.n A; most preferably Y is N(CH.sub.2).sub.n A.

Suitably, Z is nitrogen, N(CH.sub.2).sub.n A or oxygen; preferably Z is nitrogen or N(CH.sub.2).sub.n A; most preferably Z is nitrogen.

Suitably, n is 4 to 12, preferably 4 to 8 and most preferably 7 or 8.

Suitably, A is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, OH, Br, cyano, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl; preferably A is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, for example CO.sub.2 C.sub.1-4 alkyl such as CO.sub.2 CH.sub.3 or CO.sub.2 C.sub.2 H.sub.5.

Suitably, A' is CO.sub.2 H or a group hydrolysable to CO.sub.2 H, 5-tetrazolyl, SO.sub.3 H, P(O)(OR).sub.2, P(O)(OH).sub.2, or P(O)(R)(OR) in which R is hydrogen or C.sub.1-4 alkyl; preferably A is CO.sub.2 H or a group hydrolysable to CO.sub.2
H, for example CO.sub.2 C.sub.1-4 alkyl such as CO.sub.2 CH.sub.3 or CO.sub.2 C.sub.2 H.sub.5.

Suitably, R.sub.1 is hydrogen, C.sub.1-4 alkyl, optionally substituted phenyl or optionally substituted heteroaryl. Preferably R.sup.1 is hydrogen.

Suitable R.sup.2 substituent or substituent for R.sup.1 as an optionally substituted phenyl groups Ar and R.sup.1 include, for example, 1 to 3 groups which may be the same or different and are selected from C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, such as CF.sub.3, halogen, hydroxy and C.sub.1-4 alkoxy.

Suitable heteroaryl groups include, for example, saturated or unsaturated 5- or 6-membered rings comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulphur. Preferably such rings include, for example, thienyl and furyl rings.

Particularly preferred compounds of structure (IV) include:

1-(8-Bromooctyl)-4,5-diphenyl-1,2,3-triazole;

2-(8-Bromooctyl)-4,5-diphenyl-1,2,3-triazole;

1-(8-cyanooctyl)-4,5-diphenyl-1,2,3-triazole;

2-(8-cyanooctyl)-4,5-diphenyl-1,2,3-triazole;

2-(8-carboxyoctyl)-4,5-diphenyl-1,2,3triazole;

1-(8-carboxyoctyl)-4,5-diphenyl-1,2,3triazole;

2-(8-ethoxycarbonyloctyl)-4,5-diphenyl-1,2,3-triazole;

2-(6-Ethoxycarbonylhexyl)-4,5-diphenyl-1,2,3-triazole;

2-(6-Carboxyheptyl)-2,4,5-triphenyl-1,2,3-triazole;

2-(7-Carboxyheptyl)-4,5-diphenyloxazole;

1-(7-bromoheptyl) -4,5-diphenyloxazole;

2-(7-cyanoheptyl)-4,5-diphenyloxazole;

8-(3,4-Diphenylpyrazol-1-yl)octanoic acid;

8-(4,5-Diphenylpyrazol-1-yl)octanoic acid;

1-(7-Methoxycarbonylheptyl)-4,5-diphenyl-1,2,3-triazole;

2-(7-Methoxycarbonylheptyl)-4,5-diphenyl-1,2,3-triazole;

1-(7-Carboxyheptyl)-4,5-diphenyl-1,2,3-triazole;

8-(3,4-diphenylpyrazol-1-yl)octanoic acid;

8-(4,5-diphenylpyrazol-1-yl)octanoic acid; and

2-(9-Hydroxynonyl)-4,5-diphenyl-1,2,3-triazole.

Preferred compounds of structure (IV) include:

1-(8-Bromooctyl)-4,5-diphenyl-1,2,3-triazole;

2-(8-cyanooctyl)-4,5-diphenyl-1,2,3-triazole;

8-(3,4-diphenylpyrazol-1-yl)octanoic acid;

2-(9-Hydroxynonyl)-4,5-diphenyl-1,2,3-triazole;

2-(7-Methoxycarbonylheptyl)-4,5-diphenyltriazole;

8-(3,4-Diphenylpyrazol-1-yl)octanoic acid;

8-(4,5-Diphenylpyrazol-1-yl)octanoic acid;

2-(6-Carboxyheptyl)-2,4,5-triphenyl-1,2,3-triazole; and

2-(7-Carboxyheptyl)-4,5-diphenyloxazole.

Most preferred compounds of structure (IV) include:

2-(9-Hydroxynonyl)-4,5-diphenyl-1,2,3-triazole;

2-(7-Methoxycarbonylheptyl)-4,5-diphenyltriazole; and

1-(8-Bromooctyl)-4,5-diphenyl-1,2,3-triazole.

The compounds of structure (IV) can be prepared using procedures analogous to those known in the art. The present invention therefore provides in a further aspect a process for the preparation of compounds of structure (IV) which comprises: ##STR12## in which R.sup.2, X, m are as described for structure (IV) and Y.sub.a is N or C(CH.sub.2).sub.n A; with a compound of structure:

in which n and A are as described for structure (IV) and L is a leaving group, or

(b) reaction of a compound of structure (IVc): ##STR13## in which R.sup.2, m and X are as described in structure (IV), Y.sub.b is N, N(CH.sub.2).sub.n A.sub.b or C(CH.sub.2).sub.n A.sub.b, Z.sub.b is N, O or N(CH.sub.2).sub.n A.sub.b provided that:

X, Y.sub.b and Z.sub.b are not all nitrogen,

when X is CR.sup.1, Y.sub.b and Z.sub.b are not both nitrogen,

when Y.sub.b is N(CH2).sub.n A.sub.b, Z.sub.b is nitrogen, and

when Z.sub.b is O, Y.sub.b is --C(CH.sub.2).sub.n A.sub.b ;

A.sub.b is a group convertible to a group A as described in structure (IV), with a reagent suitable to convert the group A.sub.b into a group A and, optionally thereafter, converting one group A into another group A, and optionally forming a salt.

Suitable leaving groups L will be apparent to those skilled in the art and include, for example, halogen, such as bromine.

Suitable groups A.sub.b convertible to a group A include, for example, where A is CO.sub.2 H, CN groups, which can be converted into CO.sub.2 H groups by reaction with, for example, sulphuric acid. Other groups and suitable reagents will be apparent to those skilled in the art.

The reaction between compounds of structures (IVa) and (IVb) can be carried out in a suitable solvent in the presence of a base at a temperature of between ambient and the reflux temperature of the solvent used. For example, compounds of structure (IV) in which X and Y are both nitrogen and Z is N(CH.sub.2).sub.n CO.sub.2 R, can be prepared by reacting a compound of structure (IVa) in which X and Y.sub.a are both nitrogen with a compound of structure (IVb) in which L is bromine and A is CO.sub.2 H, in aqueous solution in the presence of sodium hydroxide as base. Further reaction of said compound of structure (IV) with, for example, p-toluene sulphonic acid in methanol gives the corresponding compound in which A is CO.sub.2 CH.sub.3. The compounds of structures (IVa) and (IVb) are available commercially, or can be prepared by standard techniques.

The reaction between compounds of structure (IVc) and a reagent suitable to convert the group A.sub.b to a group A will, of course, take place under conditions which will depend on the nature of the group A.sub.b. As already described, for example when A.sub.b is CN, reaction with sulphuric acid under aqueous conditions affords the desired compounds of structure (IV) in which A is CO.sub.2 H.

Other suitable groups and conditions will be apparent to those skilled in the art. Compounds of structure (IVc) are available commercially or can be prepared by standard procedures. For example, compounds of structure (IVc) in which X is nitrogen, Y.sub.b is C(CH.sub.2).sub.n CN and Z.sub.b is oxygen can be prepared via the following reaction sequence: ##STR14##

Examples 56 to 68 in the Synthetic Chemistry section serve to illustrate the preparation of compounds representative of structure (IV).

Compounds of Formula (V) are represented by the structure: ##STR15## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, or optionally substituted phenyl;

n is 2 or 4 to 12;

X is cyano, CO.sub.2 H or a group hydrolysable to CO.sub.2 H;

R.sub.3 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;

q is an integer having a value of 1 to 3;

or a pharmaceutically acceptable salt thereof.

Suitably, p is 1 to 3, and R.sub.3 is independently selected from hydrogen, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, such as CF.sub.3, halogen, hydroxy or C.sub.1-4 alkoxy. Preferably R.sub.3 is hydrogen.

Suitable when n is 2 then X is not cyano.

Suitably, R.sub.1 is hydrogen, C.sub.1-8 alkyl, C.sub.1-8 alkoxy, SC.sub.1-8 alkyl, optionally substituted phenyl, or phenyl C.sub.1-4 alkyl in which the phenyl group is optionally substituted. Preferably R.sub.1 is C.sub.1-4 alkyl or optionally substituted phenyl. When R.sub.1 is an optionally substituted phenyl the substituent include, for example, 1 to 3 groups which may be the same or different and are selected from C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, such as CF.sub.3, halogen, hydroxy and C.sub.1-4 alkoxy.

Suitably, n and m together are 4 to 12, preferably 4 to 8, and most preferably 6 or 7.

Suitable groups X, hydrolysable to CO.sub.2 H include for example, nitriles, amides and ester groups. Examples of ester groups are C.sub.1-6 alkyl esters and optionally substituted benzyl esters. Particular ester groups include mono-C.sub.1-4
alkoxycarbonyl groups such as ethoxycarbonyl and methoxycarbonyl, and tri-C.sub.1-4 alkoxy carbonyl groups such as methoxyethoxyethoxy carbonyl groups (CH.sub.3 O(CH.sub.2).sub.2 O(CH.sub.2).sub.2 O--C(O)--).

Compounds of Formula (V) include:

Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate;

Ethyl 6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;

Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;

9-[1-(3,4,5-Triphenyl-2-oxo-2,3-dihydroimidazolyl)]nonanoic acid;

7-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)heptanitrile;

Ethyl 6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;

11-(3,4,5-Triphenyl-2-oxo-1,2-dihydroimidazol-1-yl)undecanoic acid; or

Ethyl-8-(4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)octanoate.

Compounds of Formula (VI) are represented by the structure: ##STR16## wherein R.sub.1 is hydrogen, C.sub.1-4 alkyl, or optionally substituted phenyl;

n is 4 to 12;

Y is oxygen or sulfur;

X is CO.sub.2 H or a group hydrolysable to CO.sub.2 H;

R.sub.3 is independently C.sub.1-4 alkyl, halo substituted C.sub.1-4 alkyl, halogen, hydroxy or C.sub.1-4 alkoxy;

q is an integer having a value of 1 to 3;

or a pharmaceutically acceptable salt thereof.

Suitably the variables R.sub.1, R.sub.3, p, n, and X as described in Formula (V) are the same for Formula (VI).

Compounds of Formula (VI) include:

Ethyl 5-(1,4,5-triphenylimidazol-1-yl-oxy)valerate;

8-(1,4,5-Triphenylimidazol-2-yl-oxy)octanamide;

8-[1,4,5-Triphenylimidazol-2-yl-oxy]octanoic acid; or

8-[1,4,5-triphenylimidazol-2-yl-oxy]octanoic acid ammonium salt.

Additional compounds which are not encompassed by Formula(s) (I) to (VI) but are useful in this invention are listed below:

7-(3,4,5-Triphenylimidazol-1-yl-oxy)heptanitrile;

8-(2,3-Diphenylmaleimido)octanoic acid;

11-(2,3-Diphenylmaleimido)undecanoic acid;

1-(7-Ethoxycarbonyl)-4-phenylimidazole;

Methyl-7-(3,4,5-triphenyl)-2-oxo-1,2-dihydroimidazol-1-yl)-5-heptynoate;

2-[4-(3-Carboxypropoxy)phenyl]-4,5-diphenylimidazole;

1-(7-Carboxyheptyl)-2-phenylimidazole;

1-(7-Ethoxycarbonyl)-4-phenylimidazole;

1-(7-Carboxyheptyl )-2-octylthio-4,5,-diphenylimidazole;

8-(1,4,5-Triphenylimidazol-2-yl-oxy)octanamide; and the pharmaceutically acceptable salts thereof.

Preferred compounds of the Formula (V), (VI) and the additional compounds noted above are:

1-(7-Carboxyheptyl)-2-octylthio-4,5,-diphenylimidazole;

8-[1,4,5-Triphenylimidazol-2-yl-oxy]octanoic acid;

Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;

Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate;

Ethyl 6-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;

7-(3,4,5-Triphenylimidazol-2-oxo-2,3-dihydroimidazol-1-yl)heptanonitrile;

Ethyl 6-(3-methyl-4,5-diphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hexanoate;

1-(7-Ethoxycarbonyl)-4-phenylimidazole; and

Methyl-7-(3,4,5-triphenyl)-2-oxo-1,2-dihydroimidazol-1-yl)-5-heptynoate.

More preferred compounds for use herein are:

1-(7-Carboxyheptyl)-2-octylthio-4,5,-diphenylimidazole;

8-[1,4,5-Triphenylimidazol-2-yl-oxy]octanoic acid;

Ethyl 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)valerate;

Ethyl 3-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)propionate; and

7-(3,4,5,-Triphenylimidazol-2-oxo-2,3-dihydroimidazol-1-yl)heptanonitrile.
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