INHIBITION OF CoA-INDEPENDENT TRANSACYLASE AND APOPTOSIS

Abstract: The present invention is to the use of inhibitors of CoA-IT for the treatment of inhibiting or reducing cell proliferation in a human or mammal by administering to a human or mammal in need thereof, an effective amount of a CoA-independent transacylase (CoA-IT) inhibitor, or a pharmaceutically acceptable salt thereof.

Description: Inhibition of CoA-independent transacylase and apoptosis FIELD OF THE INVENTION: The present invention is to the discovery of a new method to treat cell proliferation and cancer in an animal. BACKGROUND OF THE INVENTION There is an extensive literature on the effects of alkyl-lyso-phospholipids on cell proliferation and cancer. l-O-octadecyl-2-O-methyl-.sn-glycero-3-phosphocholine (ET- I8-O-CH3), along with other alkyl-lyso-phospholipids, has been shown to decrease cell proliferation of leukemia, carcinoma and sarcoma cell lines (Andreesen et al, Blood 54: 519-523 (1979); Modolell et al, Can. Res. 39: 4681-4686 (1979); Berdel et al, 1981; Hoffman et al, Blood 63: 545-552 (1984); Morris-Natschke et al, J. Med. Chem. 29: 2114-2117 (1986); Brachwitz et al, Lipids 22: 897-903 (1987); Lu et al, Can. Res. 52: 2806-2812 (1992)). This effect on proliferation appears to be selective, in that these compounds do not block proliferation of non-cancerous cells, such as normal bone marrow cells, neutrophils and skin fibroblasts (Hoffman et al, Blood 63: 545-552 (1984); Verdonck et al, Can. Res. 50: 4020-4025 (1990); Langen et al, Anticancer Res. 12: 2109-2112 (1992)). Recent studies suggest that ET-I8-OCH3 induces apoptosis in certain cell types (Mollinedo et al, Biochem. Biophys. Res. Commun. 192: 603-609 (1993)). However, the biochemical steps leading to apoptosis have not be identified. In addition to blocking cancer cell proliferation in vitro, some evidence exists that treatment of animals with ET- 18-O-CH3 and other alkyl-lyso-phospholipids can reduce the size of tumors. See Hilgard et al, Eur. J. Cancer Clin. Oncol. 24: 1457-1461 (1988); Berger et al, Lipids 22: 935-942 (1987); and Scherf et al, Lipids 22: 927-929 (1987). In addition, ET- 18-O-CH3 has shown some efficacy in the treatment of patients suffering from leukemias and carcinomas (Berdel et al, Cancer 50: 2011-2015 (1982); Andreesen et al, J. Natl. Can. Inst. 70: 623-627 (1983); Andreesen, Prog. Biochem. Pharmacol. 22: 118-131 (1988)). The mechanism of action of these anti-proliferative alkyl-lyso-phospholipids remains elusive. It is thought that the actions ascribed to ET-I8-O-CH3 are due to the parent compound itself, as little cellular metabolism of ET-I8-O-CH3 is observed (Wilcox et al, Lipids 22: 8000-807 (1987); Lu et al, Biochem. Cell Biol. 71: 122-126 (1993)). Although structurally similar to platelet-activating factor (PAF), these compounds do not appear to act through the receptor for platelet-activating factor because antagonists of this receptor do not block the anti-proliferative effects of alkyl-lyso- phospholipids (Workman et al, Biochem. Pharmacol. 41: 319-322 (1991)). Other potential mechanisms for the anti-proliferative effects of alkyl-lyso-phospholipids have been described: inhibition of phosphatidylinositol-phospholipase C with decreases in inositol-phosphate formation (IC50 ~40 μM) (Powis et al, Cancer and Metastasis Rev. 13: 91-103 (1994)); alteration of membrane-associated enzymes such as protein kinase C and Na,K- ATPase (Zheng et al, Can. Res. 50: 3025-3031 (1990); Hoffman et al, Acta Lipids Lipid Metab. 1127: 74-80 (1992)); decreases in acyl-CoA acyltransferase activity (IC50 -80 μM) (Herrmann et al, J. Biol. Chem. 17: 7742-7747 (1986)); and decreases in phospholipid metabolism (Modolell et al, Cancer Res. 39: 4681-4686 (1979); Herrmann, D. J., Natl. Can. Inst. 75: 423-430 (1985)). However, these postulated mechanisms have not satisfactorily explained all of the actions of alkyl-lyso-phospholipids and concentrations of alkyl-lyso-phospholipids necessary to inhibit the above-mentioned cellular events are higher than their anti-proliferative effects (3-10 μM). In addition, other compounds which inhibit these enzymes are not themselves anti-proliferative (Daniel et al, Lipids 22:851-855 (1987)). Finally, there are reports of a lack of correlation between decreases in phospholipid metabolism and the sensitivity of different cell lines to blockade of proliferation (Lu et al, Can. Res. 52: 2813-2817 (1992); Lu et al, Can. Res. 52: 2806-2812 (1992)). Coenzyme A-independent transacylase (CoA-IT) is an enzyme responsible for the movement of arachidonate between phospholipid molecular species of inflammatory cells. CoA-IT removes arachidonate from the sn-2 position of 1-acyl-containing phosphoiipids, such as l-acyl-2-arachidonoyl-.m-glycero-3-phosphocholine (l-acyl-2- arachidonoyl-GPC). It then transfers that arachidonate to a suitable lyso-phospholipid acceptor, such as l-alkyl-2-lyso-GPC and l-alkenyl-2-lyso-5«-glycero-3-phospho- ethanolamine (Sugiura et al, J. Biol. Chem. 262: 1199-1205 (1987); Kramer and Deykin, Biol. Chem. 258: 13806-13811 (1983); Chilton et al, J. Biol. Chem. 258: 7268-7271 (1983)). This activity is selective for 20 carbon fatty acyl groups and is the mechanism by which inflammatory cells move arachidonate into specific phospholipid pools prior to its release (Winkler and Chilton, Drug News Perspec. 6: 133-138 (1993); Snyder et al, J. Lipid Mediat. 10: 25-31 (1994)). There exists a need to find better chemotherapeutic agents which have non-toxic pharmacological and toxicological profiles for use in mammals. These compounds should block cell proliferation of diseased cells, and hence provide potential treatment for cancers, such as leukemia and other proliferative diseases and conditions, such as psoriasis. SUMMARY OF THE INVENTION The present invention is a method to decreasing, inhibiting or reducing cell proliferation in a mammal, and inducing apoptosis in a mammal, preferably in a human, by inhibition of the enzyme CoA-IT. This effect, induced by inhibition of CoA-IT, can be accomplished by structurally distinct classes of compounds. Therefore the present invention is to the use of CoA-IT inhibitors for the treatment of cell proliferation, in mammals, preferably humans, in need of such treatment, by administering to said human an effective amount of a CoA-IT inhibitory compound, or pharmaceutically acceptable salt thereof. A preferred disease state, for treatment herein, associated with cell proliferation is psoriasis, rheumatoid arthritis or atherosclerosis. Another aspect of the present invention is to a method of treating a cancerous cell growth in a mammal, preferably a human, in need of such treatment, which method comprises administering to said mammal an effective amount of a CoA-independent transacylase (CoA-IT) inhibitor, or pharmaceutically acceptable salt thereof; provided that the inhibitor is not l-O-octadecyl-2-O-methyl-sn-glycero-3-phosphocholine (ET-18- O-CH3) or an alkyl lysophospholipid analog. Alternatively, the CoA-IT inhibitor may be co-administered with an effective amount of ET-I8-O-CH3 or an alkyl lysophospholipid analog. A preferred disease state, for treatment herein, associated with cancerous cell growth is leukemia. Another aspect of the present invention is to a method of inducing apoptosis in a mammal, in need of such treatment which method comprises administering to said human or mammal an effective amount of a CoA-independent transacylase (CoA-IT) inhibitor, or pharmaceutically acceptable salt thereof; provided that the inhibitor is not l-O- octadecyl-2-O-methyl-5rt-glycero-3-phosphocholine (ET-I8-O-CH3) or an alkyl lysophospholipid analog. Alternatively, the CoA-IT inhibitor may be co-administered with an effective amount of ET-I8-O-CH3 or an alkyl lysophospholipid analog. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 demonstrates the effect of ET-I8-O-CH3 on CoA-IT activity. Microsomes from U937 cells were incubated with the indicated concentrations of ET-I8-O-CH3 for 10 minutes then CoA-IT activity has measured as described in Methods section herein. The data are the means + SD of triplicate determinations and are representative of 3 experiments. Figure 2 demonstrates the kinetics of ET-I8-O-CH3 inhibition of CoA-IT activity. U937 microsomes were incubate with the indicated concentrations of ET-I8-O-CH3 and l-alkyl-2-lyso-GPC as described in the Methods section herein. The quantities of product produced in the reactions were determined. A double-reciprocal plot of 1/velocity versus l/[product] is shown. Figure 3 demonstrates the effect of ET-I8-O-CH3 (A), diethyl 7-(3,4,5-triphenyl-2-oxo- 2,3-dihydro-imidazol-l-yl)heptane-phosphonate (B, Compound III) and 2-[2-[3-(4- Chloro-3-trifluoromethylphenyl) ureido] -4-trifluoromethyl phenoxy]-4,5- dichlorobenzene sulfonic acid (C, Compound I) on HL-60 cell proliferation. HL-60 cells were treated at time 0 with the indicated concentrations of compound and the number of viable cells determined by trypan blue exclusion after 24 and 48 hours. Figure 4 demonstrates the effect of ET-I8-O-CH3 on DNA synthesis in HL-60 cells. HL-60 cells were treated with various concentrations of compound or vehicle. Twenty- four hours prior to harvesting the cells, they were given a pulse of [^Hlthymidine. At the indicated time points, the cells were harvested, washed, lysed and the amount of [^H] incorporated into DNA was measured as described in the Methods. The results are the means + SD of triplicate determinations and are representative of 3 experiments. Figure 5 demonstrates the effects of ET-I8-O-CH3, 2-[2-[3-(4-Chloro-3- trifluoromethylphenyl)ureido]-4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid (Compound I) and diethyl 7-(3,4,5-tripheny l-2-oxo-2,3-dihydro- imidazol- 1- yl)heptane-phosphonate (Compound III) on DNA fragmentation in HL-60 cells. HL-60 cells were treated with the indicated concentration of compound and DNA fragmentation determinate after 24 hours as described in the Methods. Control is vehicle treatment and the arrow indicates the internal standard fragment. Figure 6 demonstrates the effects of ET-I8-O-CH3, 2-[2-[3-(4-Chloro-3- trifluoromethylphenyl)ureido]-4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid (Compound I) and diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-l- yl)heptane-phosphonate (Compound III) on the content of arachidonate in phosphatidylethanolamine subclasses of HL-60 cells. HL-60 cells were treated with ET- I8-O-CH3 (25 μM), Compound I (50 μM), Compound III (25 μM) or vehicle for 48 hours and A A content determined as described in the Methods. Figure 7 demonstrates the incoφoration of [3H]thymidine by chronic myelogenous leukemic cells following incubation with inhibitors for 20 hours. - o - DETAILED DESCRIPTION OF THE INVENTION It has been found that CoA-IT represents a crucial event in the regulation of 'cancer' cell growth, and in particular apoptosis, Therefore, inhibition of CoA-IT will have a profound therapeutic effect on proliferative diseases and treatment of them thereby. ET-I8-O-CH3 was examined for its effect on CoA-IT activity in U937 microsomes by measuring the acylation of l-[-1H]alkyl-2-lyso-GPC to form product, [ 3 H]l-alkyl-2-arachidonoyl-GPC. The results (Figure 1) showed that ET-I8-O-CH3 was a potent inhibitor of CoA-IT activity with an IC5 of 0.5 uM and complete inhibition observed at 3 uM. To further explore the interaction of ET-I8-O-CH3 with CoA-IT, kinetic analysis experiments were performed. These data (Figure 2) revealed that ET-18- O-CH3 inhibited CoA-IT activity in a manner that was best fit by a model of competitive competition. The apparent Km of the substrate was 0.013 + 0.005 uM and the K js of ET- I8-O-CH3 was 1.9 ± 0.9 uM. These results show that ET-I8-O-CH3 is a CoA-IT inhibitor, interacting with CoA-IT in a specific and predictable manner and that the inhibition of CoA-IT was not caused by a non-specific effect, such as a detergent effect on the microsomal membranes. As described above, ET-I8-O-CH3 has been shown to block the proliferation of cancer cells. Therefore, another aspect of the present invention is the discovery that ET- I8-O-CH3 is a potent inhibitor of the enzyme CoA-IT and hence is useful for treating a CoA-IT mediated disease or disorder in a mammal in need thereof. This method comprises administering to said mammal an effective amount of l-O-octadecyl-2-O- methyl--?n-glycero-3-phosphocholine (ET-I8-O-CH3) as a CoA-IT inhibitor; provided that the disease or disorder is other than inhibiting cell proliferation or inducing apoptosis. The findings that ET-I8-O-CH3 is a potent inhibitor of CoA-IT raised the question of whether other structurally different CoA-IT inhibitors would influence proliferation in a manner comparable to ET-I8-O-CH3. Figure 3 illustrates the effects of ET-I8-O-CH3 and two distinct classes of compounds which are CoA-IT inhibitors on the proliferation of HL-60 cells. ET-I8-O-CH3 reduced the number of HL-60 cells in culture in a dose and time-dependent manner (Figure 3). In addition, two other structurally different CoA-IT inhibitors blocked the increases in cell numbers in a comparable fashion to ET-I8-O-CH3 (Figure 3). All three inhibitors caused a loss of cell viability that was observed between 24 and 48 h. To confirm the antiproliferative effects of the CoA-IT inhibitors, the synthesis of new DNA was monitored by H] thymidine incorporation. The results depicted in Figure 4 show that ET-I8-O-CH3 caused a time and concentration-dependent reduction in DNA synthesis. Similar results are seen with 2-[2-[3-(4-chloro-3- trifluoromethylphenyl) ureido] -4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid and diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-l-yl)heptane- phosphonate. Together, these data reveal that in addition to sharing the capacity to block the enzyme CoA-IT, ET-I8-O-CH3, 2-[2-[3-(4-chloro-3-trifluoromethylphenyl) ureido] - 4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid and diethyl 7-(3,4,5- triphenyl-2-oxo-2,3-dihydro-imidazol-l-yl)heptane-phosphonate all attenuate the growth and viability of HL-60 cells. A major mechanism that has been demonstrated to reduce cell numbers is the induction of programmed cell death or apoptosis (Bergamaschi et al, Haematologica 79: 86-93 (1994)). The induction of apoptosis has been proposed to be responsible for much of the inhibition in HL-60 cell numbers observed after treatment with ET-I8-O-CH3 (Mollinedo et al, Biochem. Biophys. Res. Commun. 192: 603-609 (1993)). The next group of experiments were performed to determine if other CoA-IT inhibitors would also instigate apoptosis. Apoptosis was determined in these studies by examining internucleosomal DNA fragmentation patterns before and after treatment with CoA-IT inhibitors. HL-60 cells treated with ET-I8-O-CH3 (6 uM), diethyl 7-(3,4,5-triphenyl-2- oxo-2,3-dihydro-imidazol-l-yl)heptane-phosphonate(50 uM) and 2-[2-[3-(4-chloro-3- trifluoromethylphenyl) ureido] -4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid (50 uM) for 24 hours all showed a pattern of DNA "ladder" fragmentation (Figure 5). DNA ladder formation in HL-60 cells depended on the dose of the CoA-IT inhibitor and occurred within 3 hours and was maximal 24 hours after treatment with CoA-IT inhibitors. Treatment of neutrophils with CoA-IT inhibitors can indirectly block the mobilization of arachidonic acid and formation of eicosanoids. A group of experiments examined whether other more direct inhibitors of arachidonic acid metabolism would induce apoptosis in HL-60 cells. Inhibitors of two phospholipase A2 isotypes as well as 5-lipoxygenase and cyclooxygenase were examined and apoptosis was determined by the presence of DNA "ladders". The results of these experiments are shown in Table 1. In Table 1 HL-60 cells were treated with the indicated concentration of compound for 24 hours and DNA fragmentation was measured as described in the Methods section herein. See also, Zileuton (Carter et al, J. Pharmacol. Exp. Ther. 256: 929-937 (1991); indomethacin (Shen, T.Y., in Anti-inflammatory and anti-rheumatic drugs, ed. by K. D. Rainsford and M. R. C. Path, pp. 149-159, CRC Press, Inc., Boca Raton, FL(1985)); AA-CF3- ketone (Street et al, Biochemistry 32: 5935-5940 (1993)). Two important findings were derived from these experiments which are 1) apoptosis is not caused by inhibitors of 5-lipoxygenase, cyclooxygenase, 14 kDa phospholipase A2 or 85 kDa phospholipase A2; and 2) CoA-IT inhibitors with widely divergent structures (ET-I8-O-CH3, 2-[2-[3-(4-chloro-3-trifluoromethylphenyl)ureido]- 4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid, diethyl 7-(3,4,5-triphenyl- 2-oxo-2,3-dihydro-imidazol-l-yl)heptane-phosphonate and diethyl 7-(l,4,5-triphenyl imidazol-2-yloxy) heptane phosphonate ) induce apoptosis. One aspect of the present invention is the demonstration that inhibition of CoA-IT has a role in apoptosis, and that inhibitors of CoA-IT, such as ET-I8-O-CH3 , 2-[2-[3-(4- chloro-3 -trifluoromethylphenyl) ureido] -4-trifluoromethyl phenoxy]-4,5- dichlorobenzene sulfonic acid and diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol- l-yl)heptane-phosphonate all caused apoptosis of HL-60 cells. To assure that something about the chemical structures of 2-[2-[3-(4-chloro-3-trifluoromethylphenyl) ureido] -4- trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid and diethyl 7-(3,4,5- triphenyl-2-oxo-2,3-dihydro-imidazol-l-yl)heptane-phosphonate, other than their capacity to inhibit CoA-IT was not causing apoptosis, close structural analogs (not having CoA-IT inhibitory activity) were tested. These latter compounds (Compounds II, V and VI; 2-[2-[3,5-bis(trifluoromethyl)phenyl]-ureidol-4-trifluoromethylphenoxy]-5-( 1,1- dimethylpropyl) benzenesulfonamide; 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-l- yl)valeric acid; and ethyl 6-(l,4,5-triphenyl imidazol- 1-yloxy) hexanoate) did not induce apoptosis suggesting that one of the key processes regulated by CoA-IT is cellular apoptosis. Arachidonate remodeling itself appears to be influencing apoptosis. To confirm this supposition, we measured the quantities of arachidonate in phosphatidylethanolamine and 1 -ether-linked phosphatidylcholine 24 and 48 hours after treatment with CoA-IT inhibitors. The enzyme CoA-IT orchestrates the movement of arachidonate into these phosphoiipids (Chilton et al, Biochemistry 34: 5403-5410 (1995)). There was a time- dependent loss of arachidonate from phosphatidylethanolamine which correlated with apoptosis in HL-60 cells (Figure 6). In addition to the loss from phosphatidyl- ethanolamine, CoA-IT inhibitors also blocked the incorporation of arachidonate into 1- ether-linked phosphatidylcholine . It has long been recognized that many 'cancer' cells contain high levels of 1 -ether- linked phosphoiipids and in particular 1 -alkenyl phosphatidylethanolamine (Snyder et al, Can. Res. 28: 972-978 (1968); and Snyder et al, Can. Res. 29: 251-257 (1969)). Moreover, Chabot et al. (Cancer Res. 49: 4441-4445 (1989)) ' have shown that susceptibility to ET- 18-O-CH3 correlates with the ether lipid content of leukemia cell lines. Extending these studies to human leukemic patients, they showed that there is a clinical correlation between response to treatment and the size of 1 -ether-linked phospholipid pools in leukemia cells (Chabot et al, Can. Res. 50: 7174-7178 (1990)). It may have been that blocking CoA-IT prevents the "loading" of AA into 1 -ether-linked phospholipid pools. The evidence in Figure 6 showing that CoA-IT inhibitors dramatically reduce arachidonate content in 1 -ether-linked phosphatidylethanolamine and phosphatidylcholine strongly support this contention. The following diagram illustrates these points. The normal path for incoφoration of arachidonate into 1 -ether-linked phosphoiipids is for arachidonate acid to be converted to arachidonate-CoA, to be then incoφorated into 1-acyl-linked phosphoiipids, such as l-acyl-2-arachidonate-phosphocholine. The enzyme CoA-IT then moves the arachidonate portion from the 1-acyl-linked phospholipid into 1 -ether-linked phosphoiipids. These 1 -ether-linked phosphoiipids are high in cancer cells. CoA-IT inhibitors block the movement of arachidonate into these 1 -ether-linked phosphoiipids, depriving cancers cells of a key component for their survival. Thus inhibition of CoA-IT induces apoptosis of cancer cells, thus dramatically influencing their growth and proliferation. Arachidonic acid Arachi dd ♦oornate -CoA 1-acyl-2-arac Jhidonate-phospholipids arachidonate in 1 -ether-linked phosphoiipids (such as: 1-alkyl-2-arachidonate-phosphocholine 1-alkenyl-2-arachidonate-phosphoethanolamine) Large amounts of 1 -ether-linked phosphoiipids in cancer cells In summary, the data support the discovery, detailed herein, that inhibition of CoA-IT activity results in a decrease in cell growth and proliferation, in induction of apoptosis, with therapeutic utility in proliferative and cancerous disorders. This invention therefore provides a method, CoA-IT inhibition, for treating proliferative and cancerous disorders. As used herein, the terms "alkyl lysophospholipid" (ALP), "alkyl lysophospholipid analog" and "ether-linked phospholipid and analogs thereof are well known terms to those of skill in the art and need no additional definitions. Generally these terms will encompass compounds that are ether linked phosphoiipids with alkyl chains in the sn- 1 position of the glycerol backbone. These compounds may be derivatives of l-O-alkyl-2-O-methyl-rac-glycero-3-phosphocholine, derivatives of l-O- hexadecyl-2-acetal-sn-glycero-3-phosphocholine (PAF), derivatives of ester- lysophospholipid (2-PLC), or derivatives of lysophophatidylcholine (2-LPC). Ether- lysophospholipids are also referred to as alkyllysoPC's, or ALPs; alkylphosphocholines are also referred to as APCs; a hexadecyl phosphocholine is referred to as HPC. Suitable compounds encompassed by this term, include but are not limited to, those described in Schick et al., Lipids, 22 (11) 904-910 (1987); such as those described in Morris- Natschke et al., J. Med. Chem., 29 (10) 2114-2117 (1986); Goto et al., Anticancer Research, 14:357362 (1994); Langen et al., Anticancer Research, 12:2109-2112 (1992); Volger et al., Lipids, 28(6) 511-516 (1993); Workman et al., Biochem. Pharm., 41(2) 319-22 (1991); Kohler et al., Inflammation, 17(3), 245-261 (1993); Brachwitz et al., Lipids, 22 (11) 897-903 (1987); Berdel, W., Lipids, 22(11) 970-973 (1987); Danhauser et al., Lipids, 22(11) 911-915 (1987); Danhauser-Riedl et al., J. Lipid Metabolism, 2, 271- 280 (1990); Lohmeyer et al., Biochem. Pharm., 45(1) 77086 (1993); Ries, et al., Chemistry and Physics of Lipids, 61, 225-234 (1992); as well as derivatives having sulfur and nitrogen linkages, such as Guivisdalsky et al., J. Med. Chem., 33 (9), 2614-2621 (1990); and Pignol et al., Anti-Cancer Drugs, 3, 599-608 (1992) whose disclosures are incoφorated by reference in their entirety herein. In addition to the compounds noted herein in the Synthetic Chemistry section, other compounds, which are CoA-IT inhibitors useful in the present invention, can be found in USSN 08/252,717; USSN 08/252,712; USSN 08/252,714; USSN 08/252,715; USSN 08/252,716; USSN 08/252,718; USSN 08/252,851; 08/102,877; 08/254,564 and its published PCT application WO/93/ 16674 whose disclosures are all incoφorated herein by reference in their entirety. TABLE 1: Ability of compounds to induce apoptosis in HL-60 cells. a 2-[2-[3-(4-chloro-3-trifluoromethylphenyl) ureido] -4-trifluoromethyl phenoxy]-4,5- dichlorobenzene sulfonic acid ^ 2-[2-[3,5-bis(trifluoromethyl)phenyl]ureidol-4-trifluoromethylphenoxy]-5-(l,l- dimethylpropyl) benzenesulfonamide c Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol- l-yl)heptane-phosphonate d Diethyl 7-(l,4,5-triphenyl imidazol-2-yloxy) heptane phosphonate e 5-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-l-yl)valeric acid f Ethyl 6-( 1 ,4,5-triphenyl imidazol- 1 -yloxy ) hexanoate 8 2-[2-[3,5-bis(trifluoromethyl) phenylsulfonamidol]-4-trifluoromethylphenoxy] benzoic acid MATERIALS AND METHODS Materials l-[3H]alkyl-2-lyso-GPC (30-60 Ci mmol) was purchased from New England Nuclear (Boston, MA). Common laboratory chemicals were purchased from Sigma Chemical Co. (St. Louis, MO). l-alkyl-2-lyso-GPC was purchased from Biomol (Plymouth Meeting, PA). Silica gel G plates were from Analtech Inc. (Newark, DE). Essentially fatty acid-free bovine serum albumin was obtained from Calbiochem (San Diego, CA). Compounds ET-I8-O-CH3 was obtained from Biomol and was made as a 100 mM stock in CHCI3, then diluted in RPMI-1640 (1:>16,000) prior to addition to cell suspensions. The following compounds were made in accordance with the synthesis as described herein: Diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-l-yl)heptane-phosphonate, as described herein ; Diethyl 7-(l,4,5-triphenyl imidazol-2-yloxy) heptane phosphonate, 5- (3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-l-yl)valeric acid, Ethyl 6-(l,4,5-triphenyl imidazol- 1-yloxy) hexanoate, 2-[2-[3,5-bis(trifluoromethyl)phenyl]ureidol-4- trifluoromethylphenoxy]-5-(l,l-dimethylpropyl) benzenesulfonamide, (2-[2-[3,5- Bis(trifluoromethyl) phenylsulfonamidol]-4-trifluoromethylphenoxy] benzoic acid sodium salt), and Zileuton. 2-[2-[3-(4-chloro-3-trifIuoromethylphenyl)ureido]-4- trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid and was obtained from Bader Chemical Co. Indomethacin was obtained from Sigma Chemicals. Arachidonyl- trifluoromethyl ketone was purchased from Cayman Chemicals, (Ann Arbor, MI). These compounds were made as stocks (5-100 mM) in dimethylsulfoxide (DMSO) and diluted in DMSO to provide final concentrations, with DMSO concentrations ranging from 0.1-1%. TNF (Sigma) was made as stock in distiller water and diluted to provide the desired concentrations. Preparation of cells HL-60 cells were obtained from American Type Culture Collection and grown in RPMI- 1640 media supplemented with 10% fetal bovine serum at 37°, 5% CO2. Cells were seeded in T-flasks at 0.03 to 0.08 x 10^ cells / ml and used for experiments at 0.5 to 0.6 x lθ6 cells / ml. The cell concentration at the time of drug exposure was found to be important parameter (Fujiwara et al, 1994). Proliferation and apoptosis The functional status of cells was measured in three ways. First cell viability was assessed by the ability of the cells to exclude trypan blue stain as shown in Figure 3. A second, more sensitive and quantitative method was to measure DNA synthesis, using the incoφoration of [^HJthymidine into newly-formed DNA as the index of synthesis. In these assays, HL-60 cells were treated with various concentrations of compounds or vehicle and kept in cell culture for different lengths of time (1 to 3 days). Cells were then given a pulse of [-^HJthymidine (1 μCi) then removed from culture 24 hours later, washed 2X with PBS and the treated with 0.2 N NaOH for 30 min followed by addition of 15% trichloroacetic acid for 2 hrs. The total sample from each well was filtered through GF/C Watman glass microfilters under vaccum and washed 3X with 5% trichloroacetic acid. The incoφoration of [*-*H]thymidine into DNA bound to the filter was quantatied using liquid scintillation spectroscopy; see Figure 4 herein. The final method, measurement of DNA fragmentation, determines not only the amount of cell death, but also provides insight into mechanism. When cells undergo programmed cell death or apoptosis, but not necrosis, cellular DNA is broken down into oligo-nucleosomal fragments which can be discerned on gels (Kerr et al, Cancer 73: 2013-2026, (1994)). Cells were treated for various times (3 - 24 hours) with different concentrations of compounds or vehicle. The washed cells were lysed by resuspending them in 200 μL of cold, sterile detergent buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA, 0.2% Triton X-l 00) and incubated on ice 30 min. Cellular protein and RNA were enzymatically degraded by incubation with 75 μg / mL RNAse A for 1 hour at 37° , then 200 μg/mL Proteinase K and 0.5% SDS (final) for 1 hour at 37°. An internal standard of DNA was added during the cell lysis and its recovery used as a confirmation of efficient extraction of cellular DNA fragments. The samples were extracted twice with an equal volume of cold, buffer saturated phenol, once with phenol / chloroform / isoamyl alcohol (25:24:1, v/v), and once with chloroform. NaCl (300 mM final) and cold ethanol were added, the solution was vortexed and let stand overnight. The ethanol was removed and the samples and DNA bp standards (Sigma) were run on a 1% agarose gel with TBE buffer with ethidium bromide (45 mM Tris, pH 8.0, 45 mM boric acid, 1 mM EDTA). The DNA was visualized under UV light (UVP Gel Documentation system) and the size of DNA fragments noted by comparison to molecular weight standards; see Figure 5 herein. CoA-IT activity CoA-IT activity was measured as described previously (Winkler et al, Biophys. Acta Lipids Lipid Metab. 1081: 339-346, (1991)) in a total volume of 100 μl. Microsomes (5-20 μg protein) from a 100,000 x g, 60 min centrifugation (Winkler et al, supra) were diluted in PBS with 1 mM EGTA to the desired protein concentration. The reaction was initiated by addition of l-[^H]alkyl-2-lyso-GPC (0.1 μCi/tube) and 1 μM final cold l-alkyl-2-lyso-GPC in assay buffer with 0.25 mg/ml fatty acid-poor bovine serum albumin. The reaction was run for 10 min at 37°. The reaction was stopped, the lipids extracted (Bligh and Dyer, Can. J. Biochem. Physiol. 37: 911-917, (1959)), and materials from an aliquot of the chloroform extract were separated by TLC in chloroform / methanol / acetic acid / water (50:25:8:4, v/v) and visualized by radioscanning (Bioscan). The product, l-[-1H]alkyl-2-acyl-GPC, was then scraped and quantified by liquid scintillation spectroscopy; see Figure 1 herein. The effects of ET-I8-O-CH3 on the kinetics of the CoA-IT reaction were examined. Various concentrations of ET-I8-O-CH3 (0-15 μM) were mixed with variable concentrations of substrate l-alkyl-2-lyso-GPC (0.01-3 μM). U937 microsomes containing CoA-IT activity were added and the reaction allowed to proceed for 3 minutes at 37°. The amount of product formed was determined as described above and was not more than 5% of added substrate at any concentration. Analysis of the results was performed using the method of Cleland,W., Methods Enzymol. 63: 103-138, (1979); see Figure 2 herein. Separation of phospholipid classes, subclasses and determination of arachidonate content HL-60 cells were exposed to compounds or vehicle and incubated for 48 hrs. At that time, incubation was terminated by extraction (Bligh and Dyer, Can. J. Biochem. Physiol. 37: 911-917 (1959)), the phospholipid classes in the organic phase were separated and collected by normal phase HPLC (Chilton, Methods Enzymol. 187: 157- 166 (1990)). The phosphoiipids were converted into diradylglycerols by addition of 20- 80 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 l,2-diradyl-3-acetylglycerols by incubation with acetic anhydride and pyridine (Chilton, Methods Enzymol. 187: 157- 166 (1990)). The phospholipid subclasses were separated by TLC in benzene / hexane / ethyl ether (50:45:4, v/v). The content of arachidonate in the phospholipid subclasses was determined after base hydrosysis, performed by using 2 M potassium hydroxide in 75% ethanol (30 min, 60°C). The reaction was stopped by addition of an equal amount of water and the reaction mixture was adjusted to pH 3 with HCl. Free fatty acids were extracted with ethyl ether, solvents removed under nitrogen and the free fatty acids converted to pentafluorobenzylesters and analyzed by GC/MS (Hubbard et al., Prostaglandins 22: 349-355 (1986)). Octadeuterioarachidonic acid was used as an internal standard and mole quantities determined using a standard curved obtained using known quantities of arachidonic acid; see Figure 6 herein. Protein analysis Protein concentrations were determined using the method of Bradford. M., Anal. Biochem. 72: 248-254, (1976), with reagents purchased from Bio-Rad (Hercules, CA). Human Luekemic Cell Lines.' One important question is whether or not the findings that inhibition of CoA- IT in the human leukemic cell line HL-60 would transfer into actual human leukemic cells. The present invention demonstrates that inhibition of CoA-IT by diverse, structurally unrelated compounds can produce an apoptotic response in cancer cells, taken directly from human patients. Method.- Blood was obtained from patients with myelogenous leukemia. Leukemic cells from these patients were treated with inhibitors of CoA-IT and proliferation measured by monitoring the incoφoration of [3H]thymidine into newly synthesized DNA. As shown in Figure 7, two structurally different inhibitors of CoA-IT activity inhibited proliferation of human cancer cells. In Figure 7, incoφoration of [3H]thymidine by chronic myelogenous leukemic cells is shown following incubation with the noted inhibitors for 20 hours. Leukemic cells from a human patient were incubated with vehicle (DMSO), diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydro- imidazol-l-yl)heptane-phosphonate (Compound III) or 2-[2-[3-(4-Chloro-3- trifluoromethylphenyl) ureido] -4-trifluoromethyl phenoxy]-4,5-dichlorobenzene sulfonic acid (Compound I) at the indicated concentrations. After 20 hours of treatment, the incoφoration of [3H]thymidine into DNA was determined as described in the Methods Section herein. To define the mechanism of this inhibition of proliferation, examination of the moφhology of these cells was made by electron microscopy. Electron microscopic analysis of human leukemic cells were performed, wherein the human leukemic cells were treated with vehicle (DMSO) or diethyl 7-(3,4,5-tri-phenyl-2-oxo-2,3-dihydro- imidazol-l-yl)heptane-phosphonate (Compound III) (25 μM) for 24 hours. After this treatment, cells were prepared for electronic microscopic analysis by standard methods. The cells treated with vehicle (DMSO) are active and viable cells, as demonstrated by the cellular processes, metabolic granules and multilobular, noncondensed nuclei. In contrast, cells treated with the CoA-IT inhibitor display moφhological changes typical of apoptosis, including condensation of the nuclei, decreased cell processes and appearance of apoptotic bodies. Figures 7 to 11 are described in greater detail below: To further confirm that the apoptotic process was initiated by treatment with CoA-IT inhibitors, cells were examined for the presence of DNA fragments, a hallmark of apoptosis. This was accomplished by labeling strand breads in the DNA with fluorescein dTUP with terminal transferase. Fluorecent analysis of DNA fragmentation, was performed for human leukemic cells treated with vehicle (DMSO) or diethyl 7- (3,4,5-triphenyl-2-oxo-2,3-dihydro-imidazol-l-yl)heptane-phosphonate (Compound III) (25 μM) for 24 hours. The cells were stained for DNA strand breaks using a kit from Boehringer Mannheim (Cat. No 1684795) according to the manufacture's instructions. Cells treated with vehicle (DMSO) had little to no fluorescent staining, whereas cells treated with the CoA-IT inhibitor had marked staining indicating that CoA-IT inhibition of human cancer cells induced apopotosis in those cells. METHOD OF TREATMENT For therapeutic use the compounds of the present invention will generally be administered in a standard pharmaceutical composition obtained by admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. For example, they may be administered orally in the form of tablets containing such excipients as starch or lactose, or in capsule, ovules or lozenges either alone or in admixture with excipients, or in the form of elixirs or suspensions containing flavouring or colouring agents. They may be injected parenterally, for example, intravenously, intramuscularly or subcutaneously. For parenteral administration, they are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The choice of form for administration as well as effective dosages will vary depending, inter alia, on the condition being treated. The choice of mode of administration and dosage is within the skill of the art. The compounds of the present invention, particularly those noted herein or their pharmaceutically acceptable salts which are active when given orally, can be formulated as liquids, for example syrups, suspensions or emulsions, tablets, capsules and lozenges. A liquid formulation will generally consist of a suspension or solution of the compound or pharmaceutically acceptable salt in a suitable liquid carrier(s) for example, ethanol, glycerine, non-aqueous solvent, for example polyethylene glycol, oils, or water with a suspending agent, preservative, flavouring or colouring agent. A composition in the form of a tablet can be prepared using any suitable pharmaceutical carrier(s) routinely used for preparing solid formulations. Examples of such carriers include magnesium stearate, starch, lactose, sucrose and cellulose. A composition in the form of a capsule can be prepared using routine encapsulation procedures. For example, pellets containing the active ingredient can be prepared using standard carriers and then filled into a hard gelatin capsule; alternatively, a dispersion or suspension can be prepared using any suitable pharmaceutical carrier(s), for example aqueous gums, celluloses, silicates or oils and the dispersion or suspension then filled into a soft gelatin capsule. Preferably the composition is in unit dose form such as a tablet or capsule. Typical parenteral compositions consist of a solution or suspension of the compound or pharmaceutically acceptable salt in a sterile aqueous carrier or parenterally acceptable oil, for example polyethylene glycol, polyvinyl pyrrolidone, lecithin, arachis oil or sesame oil. Alternatively, the solution can be lyophilised and then reconstituted with a suitable solvent just prior to administration. A typical suppository formulation comprises a compound or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent such as polymeric glycols, gelatins or cocoa butter or other low melting vegetable or synthetic waxes or fats. For the compounds noted herein, each dosage unit for oral administration contains from about 1 to 250 mg (and for parenteral administration contains preferably from 0.1 to 25 mg) of the compound, or a pharmaceutically acceptable salt thereof calculated as the free base. The pharmaceutically acceptable compounds of the invention will normally be administered to a subject in a daily dosage regimen. For an adult patient this may be, for example, an oral dose of between 1 mg and 500 mg, preferably between 1 mg and 250 mg, or an intravenous, subcutaneous, or intramuscular dose of between 0.1 mg and 100 mg, preferably between 0.1 mg and 25 mg, of the compound, or a pharmaceutically acceptable salt thereof calculated as the free base, the compound being administered 1 to 4 times per day. Disease states which could benefit from the inhibition of cell proliferation include, but are not limited to, rheumatoid arthritis, psoriasis, atherosclerosis, and cancers such as leukemia or solid tumors. SYNTHETIC CHEMISTRY Without further elaboration, it is believed that one skilled in the art can, using the preceding descriptions, utilize the present invention to its fullest extent. The following examples further illustrate the synthesis of compounds of this invention. The following examples are, therefore, to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. Temperatures are recorded in degrees centigrade unless otherwise noted. Example 1 Preparation of 2-r2-r3.5-Bis(trifluoromethyDphenylsulfonamidol-4-trifluoromethyl- phenoxylbenzoic acid a) Methyl 2-(2-nitro-4-trifluoromethylphenoxy)benzoate A mixture of methyl salicylate (15.2 g, 0.1 mol), 4-chloro-3-nitrobenzotrifluoride (22.6 g, 0.1 mol), and potassium carbonate (8.3 g, 0.06 mol) in dimethylformamide (100 mL) was stirred under argon and heated in an oil bath to 150 β C for 70 min. The reaction mixture was diluted with ethyl acetate (300 mL), filtered, and the residue washed with ethyl acetate. The combined filtrates were evaporated, and the crude product was purified by flash chromatography (silica gel, ethyl acetate/hexane) to give the title compound as a pale yellow solid; mp 69-70°C. b) Methyl 2-(2-amino-4-trifluoromethylphenoxy)benzoate A mixture of methyl 2-(2-nitro-4-trifluoromethylphenoxy)benzoate (26 g, 0.076 mol) and 10% palladium on carbon (1 g) in ethyl acetate (400 mL) was hydrogenated in a parr bottle at 55 psi for 2 h. The reaction mixture was flushed with argon, filtered through Celite® and evaporated. The crude product was purified by flash chromatography (silica gel, ethyl acetate/hexane) to give the title compound as a white solid. --H NMR (250 MHz, CDCI3) d 7.91 (dd, IH), 7.45-7.53 (m, IH), 7.17-7.24 (m, IH), 6.88-7.03 (m, 3H), 6.78 (d, IH), 4.25 (br s, 2H), 3.84 (s, 3H). c) 2-(2-Amino-4-trifluoromethylphenoxy)benzoic acid A mixture of methyl 2-(2-amino-4-trifluoromethylphenoxy)benzoate (19.2 g, 0.062 mol) and 2 N sodium hydroxide (65 mL, 0.13 mol) in tetrahydrofuran (100 mL) was warmed to 50°C while stirring under argon for 3 h. The tetrahydrofuran was evaporated, and the aqueous solution was acidified with dilute HCl, extracted with ethyl acetate, dried over anhydrous MgSO4 ; filtered and evaporated. The crude product was recrystallized from EtOAc/hexane to give the title compound as a white crystalline solid. 1H NMR (250 MHz, CD3OD) d 7.91 (d, IH), 7.50 (dd, IH), 7.22 (dd, IH), 7.11 (d, IH), 6.97 (d, IH), 6.86 (d, IH), 6.78 (d, IH). d) Benzhydrol 2-(2-amino-4-trifluoromethylphenoxy)benzoate A mixture of 2-(2-amino-4-trifluoromethy Iphenoxy )benzoic acid (5 g, 0.017 mol) in EtOAc (50 mL) was stirred under argon at 50°C. A solution of diphenyldiazomethane (3.3g, 0.017 mol) dissolved in toluene (50 mL) was added dropwise. The solvents were evaporated and the residue was purified by flash chromatography (silica gel, ethyl acetate/hexane) to give the title compound as a viscous oil. 1H NMR (250 MHz, CDCI3) _ 8.02 (dd, IH), 7.45-7.52 (m, IH), 7.20-7.33 (m, I IH), 7.07 (s, IH), 6.96-7.07 (m, 2 H), 6.86-6.91 (m, IH), 6.73 (d, IH), 4.00 (br s, 2H). e) Benzhydrol 2-[2-[3,5-bis (trifluoromethyl)phenylsulfonamido]-4- trifluoromethylphenoxy]benzoate Benzhydrol 2-(2-amino-4-trifluoromethylphenoxy)benzoate (4 g, 8.6 mmol) was dissolved in pyridine (25 mL) and stirred under argon at room temperature. 3,5- bis(trifluoromethyl)benzenesulfonyl chloride (4.05 g, 12.9 mmol) was added in portions, and the mixture stirred for 16 h. The solvent was evaporated, and the residue was purified by flash chromatography (silica gel, ethyl acetate/hexane) to give the title compound as a white solid. H NMR (250 MHz, CDCI3) d 8.25 (br s, IH), 8.15 (s, 2H), 7.99 (dd, IH), 7.85-7.92 (m, 2H), 7.1-7.5 (m, 13H), 7.01 (s, IH), 6.72 (d, IH), 6.45 (dd, 1 H). f) 2-[2-[3,5-Bis (trifluoromethyl)phenylsulfonamido]-4-trifluoromethylphenoxy]benzoic acid Benzhydrol 2-[2-[3,5-bis (trifluoromethyl)phenylsulfonamido]-4- trifluoromethylphenoxyjbenzoate (3.7 g, 5 mmol) was dissolved in ethyl acetate (125 mL) and was hydrogenated for 0.75 hour at 50 psi over 10% palladium on carbon (0.5g). The catalyst was filtered off and the solvent evaporated to give the crude product which was purified by flash chromatography (silica gel, ethyl acetate/hexane/formic acid). Recrystallization from ethyl acetate/hexane gave the title compound as a white crystalline solid; mp 133-134°C. In a similar manner, the sodium salt of 2-[2-[3,5-Bis (trifluoromethyl)phenyl- sulfonamido]-4-trifluoromethylphenoxy]benzoic acid may also be prepared. EXAMPLE 2 2-r2-r3-(4-Chloro-3-(trifluoromethyl')phenynureido1-4-(trifluoromethyl')phenoxy1-4.5- dichlorobenzenesulfonic acid, sodium salt a) 2-(2-Nitro-4-trifluoromethylphenoxy)-4,5- dichlorobenzene A mixture of 3,4-dichlorophenol (25 gram (hereinafter g), 0.153 mol), 4-chloro-3- nitrobenzo-trifluoride (52 g, 0.23 mol) and potassium carbonate (63 g, 0.459 mol) in dimethylformamide (450 mL)was stirred under argon at 120°C for 24 h. The reaction mixture was filtered and the filtrate evaporated. The residue was dissolved in ethyl acetate, washed with water, brine, dried over anhydrous magnesium sulfate, filtered and evaporated. The residue was flash chromatographed (silica gel, methylene chloride/hexane), to give the title compound as a yellow oil. b) 2-(2-Nitro-4-trifluoromethylphenoxy)-4,5-dichlorobenzenesulfonic acid, ammonium salt Fuming sulfuric acid (8 mililiter (hereinafter mL) was cooled to -20°C and 2-(2- nitro-4-trifluoromethyl-phenoxy)-4,5-dichlorobenzene (8g, 22.8 mmol) in methylene chloride (8 mL) was added to the solution. The mixture was stirred for 15 min and then quenched in ice. Extraction with ethyl acetate followed by flash chromatography (silica gel, methylene chloride/isopropanol/ammonium hydroxide) gave the title compound. 1H NMR (400 MH Z , CD3OD) 8.31 (d, IH), 8.11 (s, IH), 7.87 (dd, IH), 7.4 (s, IH), 7.16 (d, IH). c) 2-(2-Amino-4-trifluoromethylphenoxy)-4,5-dichlorobenzenesulfonic acid The 2-(2-nitro-4-trifluoromethylphenoxy)-4,5-dichlorobenzenesulfonic acid, ammonium salt (5 g, 11.2 mmol) was dissolved in acetic acid (75 mL). The mixture was diluted with water (75 mL), and a 20 % aqueous solution of titanium trichloride (65 mL)was added. After aqueous workup the crude product was'flash chromatographed (silica gel, methylene chloride/ethanol/ammonium hydroxide) to give the title compound. -Η NMR (400 MH Z , CD3OD) d 8.02 (s, IH), 7.17 (d, IH), 7.12 (d, IH), 6.94 (dd, IH), 6.87 (s, IH). d) 2-[2-[3-(4-Chloro-3-(trifluoromethyl)phenyl]ureido]-4-(trifluoromethyl)phenoxy]-4,5- dichlorobenzenesulfonic acid, ammonium salt A solution of 2-(2-amino-4-trifluoromethylphenoxy)-4,5-dichlorobenzenesulfonic acid (2 g, 4.8 mmol) and 4-chloro-3-trifluoromethylphenyl isocyanate (1.11 g, 5 mmol) in pyridine (20 mL) was stirred under argon for 72 hours (hereinafter h). The solvent was evaporated and the residue was flash chromatographed (silica gel, methylene chloride/ethanol/ammonium hydroxide) to give the title compound. -Η NMR (400 MH Z; CD3OD) d 8.61 (d, IH), 8.08 (s, IH), 7.95 (d, IH), 7.60 (dd, IH), 7.43 (d, IH), 7.30- 7.36 (m, 2H), 7.10 (s, IH). e) 2-[2-[3-(4-Chloro-3-(trifluoromethyl)phenyl]ureido]-4-(trifluoromethyl)phenoxy]-4,5- dichlorobenzenesulfonic acid, sodium salt A mixture of 2-[2-[3-(4-chloro-3-(trifluoromethyl)phenyl]ureido]-4-(trifluoro- methyl)phenoxy]-4,5-dichlorobenzenesulfonic acid, ammonium salt (2.1 g, 3.28 mmol) and sodium bicarbonate (0.33 lg, 3.94 mmol) in methanol (20 mL) and water (2 mL) were stirred under argon for 30 min. The solvents were evaporated, and the residue was flash chromatographed (Ci8 reverse phase, MeOH/H2θ) to give the title compound after lyophilization. MS (ES) m/e 643.8 [M+H]+. EXAMPLE 3 Diethyl 7-(3.4.5-triphenyl-2-oxo-2.3-dihydro-imidazol- 1 -v heptane-phosphonate A solution of l,4,5-triphenyl-3-(7-bromoheptyl)imidazol-2-one (l.Og) and triethylphosphite (1.66g) in xylene (5ml) was heated at reflux temperature for 40 hours. The solution was evaporated to an oil and chromatographed on silica gel (ethyl acetate/ethanol). The resulting oil was taken up in diethyl ether, filtered and evaporated to give diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-l-yl)heptanephosphonate as a clear oil (0.84g, 75%). Found: C, 70.11 ; H, 7.37; N, 4.94%; C 32 H 39 N 2 O 4 P; Requires: C, 70.31; H, 7.19; N, 5.12% EXAMPLE 4 Diethyl 7-d AS-triphenylimidazol-Σ-yloxylheptanephosphonate A mixture of l,4,5-triphenyl-3-(7-bromoheptyl)-imidazol-2-one (l.Og) and triethyl phosphite (1.66g) was heated at reflux temperature in xylene (5 ml) for 40 hours. The solution was evaporated to an oil and re-evaporated from ethanol. The oil was partitioned between ether and water, the ether solution was separated, dried over magnesium sulphate and evaporated to an oil which was chromato-graphed on silica gel (ethyl acetate) to give diethyl 7-(l,4,5-triphenylimidazol-2-yl-oxy)heptanephosphonate as an oil which solidified on standing to a white solid (0.83g; 75%), m.p. 76-7°C. Found: C, 70.49; H, 7.40; N, 4.94%; C 32 H 39 N 2 O 4 P; Requires: C, 70.31; H, 7.19; N, 5.12%. EXAMPLE 5 Ethyl 6-( 1.4.5-triphenylimidazol-2-oxy)hexanoate A mixture of ethyl-6- bromohexanoate (13.38g), 1,4,5-triphenylimidazole (6.24g), potassium carbonate (13.82g) and butan-2-one (250ml) was stirred under nitrogen at reflux for 24 hours. The mixrture was filtered and the filtrate was evaporated. The residue was chromatographed on silica gel eluted with 10% ethyl acetate to give ethyl 6-(l,4,5-triphenylimidazol-2-one- 3-yl)hexanoate (5.61g) and the title compound, (2.21g) m.p. 102 - 103.5°C. Found: C, 76.73; H, 6.73; N, 6.11%; (C29H30N2O3) requires: C, 76.63; H, 6.65; N, 6.16% The compound may also be synthesized in accordance with the procedures indicated in DE 3,228, 271 Al whose disclosure is incoφorated herein by reference in its entirety. EXAMPLE 6 5-( 1 AS-Triphenylimidazol-Σ-one-S-yllvalerate a) Ethyl 5-(l,4,5-triphenylimidazol-2-yloxy)valerate l,4,5-Triphenylimidazol-2-one was treated with ethyl 5-bromovalerate and potassium carbonate in butanone to give after chromatographic work-up ethyl 5-( 1,4,5- triphenylimidazol-2-yloxy)valerate, m.p. 95-96°C, Found: C, 76.5;H, 6.5; N, 6.3%; C28H28N2O3 requires: C, 76.3;H, 6.4;N,6.4% b) A mixture of ethyl 5-(l,4,5-triphenylimidazol-2-one-3-yl)valerate(1.25g), 10% sodium hydroxide solution and ethanol (50ml) was stirred for 1 hour. The solution was acidified and the precipitate was collected. Recrystallisation from methanol gave the title compound, (0.66g) m.p. 130-132°C. Found: C, 75.80; H, 5.91; N, 6.67%; ( C 26 H 24 N 2°3) requires: C, 75.71; H, 5.86; N, 6.79% EXAMPLE 7 Preparation of 2-r2- 3-r3.5-Bis(trifluoromethyl ' )phenyllureidol-4- trifluoromethylphenoxyl-5-( 1.1 -dimethylpropyllbenzenesulfonamide a) 2-(Nitro)-4-(trifluoromethyl)phenoxy-4-( 1 , 1 -dimethylpropyl)benzenesulfonic acid ammonium salt. 5-(l,l-dimethylpropyl)2-(hydroxyl)benzenesulfonic acid ammonium salt (l.Og, 0.00383 mol), 4-bromo-3-nitrobenzotrifluoride (1.5g, 0.00557 mol) and K2CO3 (l.Og, 0.00766 mol) were mixed in dimethylformamide (25 ml) at 150 ° C for sixteen hours. The reaction was purified by flash chromatography (silica gel, methylene chloride/isopropanol/ammonium hyroxide) to yield the title compound ( 1.0g, 58%). H NMR(250 MHz, CD3OD) d 8.22 (d, IH), 8.05 (d, IH), 7.70-7.60 (dd, IH), 7.55-7.45 (dd, IH), 7.10-7.00 (m, 2H), 1.78-1.65 (q, 2H), 1.42 (s, 6H), 0.78-0.64 (t, 3H). b) 2-Nitro-4-(trifluoromethyl)phenoxy-4-( 1 , 1 -dimethylpropyl)benzenesulfonyl chloride 2-Nitro-4-(trifluoromethyl)phenoxy-4-( 1 , l-dimethylpropyl)benzenesulfonic acid ammonium salt (500mg, 0.0011 mol) and thionyl chloride (5.0ml, 0.0688mol) were mixed for thirty minutes. Five drops of dimethylformamide were added and a color change was noted within five minutes (puφle to yellow). Reaction was allowed to mix for an hour. It was determined done by TLC. The thionyl chloride was evaporated and the title compound was used without further purification or characterization. c) 2-Nitro-4-(trifluoromethyl)phenoxy-4-( 1 , l-dimethylpropyl)benzenesulfonamide 2-Nitro-4-(trifluoromethyl)phenoxy-4-( 1 , 1 -dimethylpropyl)benzenesulfonyl chloride was mixed in ammonium hydroxide (20ml) at 0° C for an hour. The reaction mixture was extracted with ethyl acetate and the organic layer dried over MgSO4 5 filtered, and flash chromatographed (silica gel, 20% ethyl acetate/hexane) to yield the title compound (300mg, 61%). -H NMR(250 MHz, CD3OD) d 8.35 (s, IH), 8.00 (s, IH), 7.85 (d, IH), 7.65 (dd, IH), 7.19 (t, 2H), 1.73 (q, 2H), 1.38 (s, 6H), 0.76 (t, 3H). d) 2- Amino-4-(trifluoromethyl)phenoxy-4-( 1 , 1 -dimethylpropyl)benzene sulfonamide 2-Nitro-4-(trifluoromethyl)phenoxy-4-( 1 , 1 -dimethylpropyl)benzenesulfonamide (300mg, 0.0007mol) was mixed in ethyl acetate (25ml) and palladium on carbon (lOOmg) was added under argon. The mixture was hydrogenated at 58 psi for five hours, filtered through celite with methanol and chloroform and evaporated. The title compound was used without further purification or characterization. e) 2-[2-[3-[3,5-Bis(trifluoromethyl)phenyl]ureido]-4-trifluoromethylphenoxy]-5-(l,l- dimethylpropyl)benzenesulfonamide 2- Amino-4-(trifluoromethyl)phenoxy-4-( 1 , 1 -dimethylpropyl)benzenesulfonamide (360mg, 0.0009mol) and 3,5-di(trifluoromethyl)phenyl isocyanate were mixed in pyridine(lθml) for sixteen hours. The pyridine was evaporated and the reaction was flash chromatographed (silica gel, 25% ethyl acetate/hexane). The product was further purified by flash chromatography (silica gel, methylene chloride/hexane/ethyl acetate) to yield the title compound (130mg, 22%). *H .NMR(250 MHz, CD3OD) δ. 8.60 (s, IH), 8.00 (s, 3H), 7.55 (s, 2H), 7.35 (m, IH), 7.15 (d, IH), 6.95 (d, IH), 1.70 (q, 2H), 1.30 (s, 6H), 0.70 (t, 3H). Abbreviations: The abbreviations used herein are: AA, arachidonic acid, 5-8-11-14 eicosatetraenoic acid; 1-alkyl, 1-O-alkyl; 1-alkenyl, l-O-alk-l'-enyl; BSA, bovine serum albumin; CoA-IT, Coenzyme A-independent transacylase; COX, cyclooxygenase; ET- I8-O-CH3, l-O-octadecyl-2-O-methyl-sn-glycero-3-phosphocholine; GPC, ^n-glycero- 3-phosphocholine; 5LO, 5-lipoxygenase; PLA2, phospholipase A2; PBS, phosphate buffered saline; TLC, thin layer chromatography; TNF, tumor necrosis factor α. The above description fully discloses the invention including preferred embodiments thereof. Modifications and improvements of the embodiments specifically disclosed herein are within the scope of the following claims. Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. Therefore the Examples herein are to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows.

Claims: WHAT IS CLAIMED IS: 1. A method of inhibiting or reducing cell proliferation in a mammal, in need of such which method comprises administering to said human or mammal an effective amount of a CoA-independent transacylase (CoA-IT) inhibitor, or pharmaceutically acceptable salt thereof; provided that the inhibitor is not l-O-octadecyl-2-O-methyl-sn- glycero-3-phosphocholine (ET-I8-O-CH3) or an alkyl lysophospholipid analog. 2. The method according to Claim 1 wherein the CoA-IT inhibitor is co¬ administered with an effective amount of ET-I8-O-CH3 or an or an alkyl lysophospholipid analog. 3. The method according to Claim 1 wherein the mammal is afflicted with psoriasis, rheumatoid arthritis, or atherosclerosis. 4. A method of treating a cancerous cell growth in a mammal, in need of such which method comprises administering to said mammal an effective amount of a CoA- independent transacylase (CoA-IT) inhibitor, or pharmaceutically acceptable salt thereof; provided that the inhibitor is not l-O-octadecyl-2-O-methyl-5n-glycero-3-phosphocholine (ET-I8-O-CH3) or an alkyl lysophospholipid analog. 5. The method according to Claim 4 wherein the CoA-IT inhibitor is co¬ administered with an effective amount of ET-I8-O-CH3 or an alkyl lysophospholipid analog. 6. The method according to Claim 4 wherein the mammal is afflicted with leukemia. 7. A method of inducing apoptosis in a mammal, in need of such treatment which method comprises administering to said human or mammal an effective amount of a CoA-independent transacylase (CoA-IT) inhibitor, or pharmaceutically acceptable salt thereof; provided that the inhibitor is not 1 -O-octadecyl-2-O-methyl-5«-glycero-3- phosphocholine (ET-I8-O-CH3) or an alkyl lysophospholipid analog. 8. The method according to Claim 7 wherein the CoA-IT inhibitor is co¬ administered with an effective amount of ET-I8-O-CH3 or an alkyl lysophospholipid analog. 9. Use of 1 -O-octadecyl-2-O-methyl-sn-glycero-3-phosphocholine (ET- 18-O-CH3) as a CoA-IT inhibitor. 10. A method for treating a CoA-IT mediated disease or disorder in a mammal in need thereof which method comprises administering to said mammal an effective amount of l-O-octadecyl-2-O-methyl-jn-glycero-3-phosphocholine (ET-I8-O-CH3) as a CoA-IT inhibitor; provided that the disease or disorder is other than inhibiting cell proliferation or inducing apoptosis.

Inventors: CHILTON FLOYD III, WINKLER JAMES DAVID, CHILTON, FLOYD, III, WINKLER, JAMES, DAVID
Applicants: CHILTON FLOYD III, SMITHKLINE BEECHAM CORP, WAKE FORREST UNIVERSITY, WINKLER JAMES DAVID, CHILTON, FLOYD, III, SMITHKLINE BEECHAM CORPORATION, WINKLER, JAMES, DAVID
Application year: 1996
Main classification: A61K31
IPC codes: A61K31, A61K45, A61P9, A61P17, A61P29, A61P43