Tetronic and thiotetronic acid derivatives as phospholipase A.sub.2 inhibitors

The present invention is directed to certain tetronic acid derivatives having anti-inflammatory activity and to a method for using them as anti-inflammatory agents.
It is now well-established that arachidonic acid (AA) is metabolized in mammals by two distinct pathways. The metabolism of arachidonic acid by cyclooxygenase enzymes results in the production of prostaglandins and thromboxanes. The physiological activity of the prostaglandins has already been amply elucidated in recent years. It is now known that prostaglandins arise from the endoperoxides PGG.sub.2 and PGH.sub.2 by the cyclooxygenase pathway of arachidonic acid metabolism. These endoperoxides are also the precursors of the thromboxanes (Tx) A.sub.2 and B.sub.2. TxA.sub.2 is a vasoconstrictor which stimulates platelet aggregation. In the normal situation, the vasoconstrictive and platelet aggregating properties of the thromboxanes are balanced by another product arising from the endoperoxides in the cyclooxygenase pathway, prostacyclin (PGI.sub.2), which is a vasodilator with platelet aggregation inhibitory activity. In the event prostacyclin synthesis is impaired and/or platelet activation is enhanced, then thrombosis and vasoconstriction is favored. The role of prostanoids in haemostasis and thrombosis are reviewed by R. J. Gryglewski, CRC Crit. Rev. Biochem., 7, 291 (1980) and J. B. Smith, Am. J. Pathol., 99, 743 (1980). Cyclooxygenase metabolites are known to participate directly in the inflammatory response [see Higgs et al., Annals of Clinical Research, 16, 287-299 (1984)]. This is through their vasodepressor activities, participation in pain and fever and augmentation of peptide mediator vascular permeability and edema forming properties. Finally, various aspects of cell mediated immunity are influenced by cyclooxygenase products.
The other pathway of AA metabolism involves lipoxygenase enzymes and results in the production of a number of oxidative products called leukotrienes. The latter are designated by the LT nomenclature system, and the most significant products of the lipoxygenase metabolic pathway are the leukotrienes B.sub.4, C.sub.4 and D.sub.4. The substance denominated slow-reacting substance of anaphylaxis (SRS-A) has been shown to consist of a mixture of leukotrienes, with LTC.sub.4 and LTD.sub.4 as the primary products and having varying amounts of other leukotriene metabolites [see Bach et al., J. Immun., 215, 115-118 (1980); Biochem. Biophys. Res. Commun., 93, 1121-1126 (1980)].
The significance of these leukotrienes is that a great deal of evidence is accumulating showing that leukotrienes participate in inflammatory reactions, exhibit chemotactic activities, stimulate lysosomal enzyme release and act as important factors in the immediate hypersensitivity reaction. It has been shown that LTC.sub.4 and LTD.sub.4 are potent bronchoconstrictors of the human bronchi [see Dahlen et al., Nature, 288, 484-486 (1980)], and another leukotriene, LTB.sub.4, is a powerful chemotactic factor for leukocytes [see A. W. Ford-Hutchinson, J. Roy Soc. Med., 831-833 (1981)]. The activity of leukotrienes and slow-reacting substances as mediators of inflammation and hypersensitivity is extensively reviewed in Bailey and Casey, Ann. Reports Med. Chem., 17, 203-207 (1982).
Phospholipase A.sub.2 (PLA.sub.2) is the critical rate limiting enzyme in the arachidonic acid (AA) cascade since it is responsible for the hydrolysis of esterified AA from the C-2 position of membrane phospholipids. This reaction generates two products (1) free AA which is then available for subsequent metabolism by either the cyclooxygenase or lipoxygenase enzymes and (2) lysophospholipid. When alkylarachidonoyl-glycerophosphatidylcholine is acted upon by the PLA.sub.2 the generation of platelet activating factor (PAF) is initiated; PAF is pro-inflammatory in its own right [see Wedmore et al., Br. J. Pharmacol., 74, 916-917 (1981)]. In this regard it may be noted that the anti-inflammatory steroids are thought to inhibit eicosanoid synthesis by inducing the synthesis of a PLA.sub.2 inhibitory protein denominated macrocortin or lipomodulin [see Flower et al., Nature, London 278, 456 (1979) and Hirata et al., Proc. Natn. Acad. Sci., U.S.A., 77, 2533 (1980)].
As the initial step leading to subsequent conversion of AA to the various eicosanoids by the cyclooxygenase and lipoxygenase pathways, the PLA.sub.2 -mediated release of AA from membrane phospholipids is a critical event in attempting to deal with the various physiological manifestations which are based on the activity of the eicosanoids and/or PAF. Thus, while PLA.sub.2 has been shown to be required for platelet aggregation [Pickett et al., Biochem. J., 160, 405 (1976)], cardiac contraction and excitation [Geisler et al., Pharm. Res. Commun., 9, 117 (1977)], as well as prostaglandin synthesis [Vogt, Adv. Prostagl. Throm. Res., 3, 89 (1978)], the inhibition of PLA.sub.2 is indicated in the therapeutic treatment of both PAF induced or cyclooxygenase and/or lipoxygenase pathway product-mediated physiological conditions. Thus, PLA.sub.2 inhibitors are a rational approach to the prevention, removal or amelioration of such conditions as allergy, anaphylaxis, asthma and inflammation.
The invention provides novel compounds of the formula ##STR4## wherein X is --CH.sub.2 R;
R is ##STR5## and further when Y=S, R may also be --(CH.sub.2).sub.e CH.sub.3 ; Y is --O-- or --S--;
R.sup.1 and R.sup.2 are each, independently, hydrogen or lower alkyl;
R.sup.3 is indolyl, furyl, phenyl or phenyl optionally mono- or disubstituted independently by alkyl of 1-7 carbon atoms, --C(CH.sub.3).sub.3, --C(CH.sub.3).sub.2 CH.sub.2 C(CH.sub.3).sub.3, --C(CH.sub.3).sub.2 CH.sub.2 CH.sub.3, haloloweralkyl, perfluoroalkyl, lower alkoxy, aryl alkoxy, halo or nitro;
R.sup.4 and R.sup.5 are, independently, --COCH.sub.2 R.sup.7, --CO.sub.2 R.sup.7, --CONHR.sup.7, geranyl or CH.sub.2 R.sup.3 ;
R.sup.6 is hydrogen or lower alkyl;
R.sup.7 is geranyl and any moiety selected from R other than ##STR6## A and B are, independently, --O--, --S-- or --NR.sup.6 --; and a is 0-8;
b is 1-10 when Y=S, and 2-10 when Y=O;
c is 1-3;
d is 0-9; and
e is 3-18.
The invention further provides a method for treating immunoinflammatory conditions such as allergy, anaphylaxis, asthma and inflammation, in mammals, which comprises administering to a mammal so afflicted an effective amount of a compound having the formula ##STR7## wherein Y is --O-- or --S--;
X is --(CH.sub.2).sub.a CH.sub.3, --(CH.sub.2).sub.b Z or --(CH.dbd.CH).sub.b Z when Y=O, and --(CH.sub.2).sub.a CH.sub.3 when Y=S;
R.sup.1 and R.sup.2 are each, independently, hydrogen or lower alkyl;
Z is indolyl, furyl, phenyl or phenyl optionally mono- or disubstituted independently by alkyl of 1-7 carbon atoms, haloloweralkyl, perfluoroalkyl, loweralkoxy, aralkoxy, halo or nitro;
a is 0-20 when Y=O, and a is 1-3 when Y=S; and
b is 1-2.
The terms "lower alkyl" and "lower alkoxy" refer to moieties having 1 to 6 carbon atoms. The term "aryl" refers to aromatic moieties having 6 to 10 carbon atoms. The term "halo" refers to fluoro, bromo or chloro.
The compounds within the scope of the invention by virtue of their configuration, exhibit stereoisomerism. Accordingly, the compounds of the invention include the diastereomers, enantiomorphs, racemates and mixtures thereof.
The compounds within the scope of the invention can be prepared by a variety of synthetic routes using conventional methods. According to one preparative scheme, a suitable R-containing reactant is condensed with tetronic acid to give an acyl tetronic acid, as exemplified by the condensation of oleic acid and tetronic acid: ##STR8##
Those compounds of the invention in which R is a moiety of the formula ##STR9## can be prepared by the following procedure. 1-Penten-5-ol is silylated with a suitable silylating agent to yield an intermediate silyloxy diol, which is osmylated to yield a saturated vicinial dihydroxysilyl ether: ##STR10## The latter ether can then be condensed, for example, with a suitable alkyl halide in the presence of sodium hydride to yield mono- or bis alkylated intermediates: ##STR11## Alternatively, condensation with a carboxylic acid gives a diacylated intermediate, while with an isocyanate, gives a dicarbamoylated intermediate: ##STR12## In either case, the resulting intermediate is subjected to deprotection and oxidation to yield, as a reactant, a carboxylic acid, which can then be condensed with tetronic acid to yield the desired final products: ##STR13##
The starting materials in the above preparative sequences are all commercially available or can be prepared by conventional methods as taught in the chemical literature.
The compounds of the invention, by virtue of their ability to inhibit activity of PLA.sub.2 enzyme, are useful in the treatment of conditions mediated by products of the oxidation of arachidonic acid. Accordingly, the compounds are indicated in the prevention and treatment of such conditions as allergic rhinitis, allergic bronchial asthma and other naso-bronchial obstructive air-passageway conditions, other immediate hypersensitivity reactions, such as allergic conjunctivitis; and various inflammatory conditions such as those present in rheumatoid arthritis, osteoarthritis, tendinitis, bursitis, psoriasis (and related skin inflammation) and the like.
When the compounds within the scope of the invention are employed in the treatment of allergic airways disorders or in anti-flammatory therapy, they can be formulated into oral dosage forms such as tablets, capsules and the like. The compounds can be administered alone or by combining them with conventional carriers, such as magnesium and carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter and the like. Diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, tablet-disintegrating agents and the like may be employed. The compounds may be encapsulated with or without other carriers. In all cases, the proportion of active ingredients in said compositions both solid and liquid will be at least to impart the desired activity thereto on oral administration. The compounds may also be injected parenterally, in which case they are used in the form of a sterile solution containing other solutes, for example, enough saline or glucose to make the solution isotonic. For administration by inhalation or insufflation, the compounds may be formulated into an aqueous or partially aqueous solution, which can then be utilized in the form of an aerosol. The compounds may also be used topically and for this purpose they may be formulated in the form of dusting powders, creams or lotions in pharmaceutically acceptable vehicles, which are applied to affected portions of the skin.
The dosage requirements vary with the particular compositions employed, the route of administration, the severity of the symptoms presented and the particular subject being treated. Treatment will generally be initiated with small dosages less than the optimum dose of the compound. Thereafter the dosage is increased until the optimum effect under the circumstances is reached. In general, the compounds of the invention are most desirably administered at a concentration that will generally afford effective results without causing any harmful or deleterious side effects, and can be administered either as a single unit dose, or if desired, the dosage may be divided into convenient subunits administered at suitable times throughout the day.
The standard pharmacological procedures, which are described fully in the examples given hereafter, inter alia, determine the specificity of action of the compounds of the invention as PLA.sub.2 inhibitors as measured by their ability to inhibit the synthesis of LTB.sub.4 and TxB.sub.2 by rat glycogen -elicited polymorphonuclear leukocytes; their ability to inhibit platelet-activating factor and LTB.sub.4 biosynthesis in human neutrophils; as well as measure their ability to inhibit arachidonic acid release mediated by human and non-human source PLA.sub.2. The procedures further measure the ability of the compounds of the invention to inhibit, in vivo, the activity of exogenously administered PLA.sub.2. The pharmacological testing additionally demonstrates the ability of the compounds of the invention to inhibit, in vivo, the lipoxygenase and cyclooxygenase pathways of arachidonic-acid metabolism; and also measures the in vivo activity of the compounds as anti-inflammatory agents in the rat carrageenan paw edema assay and the murine ear edema assay.

The following examples show the preparation and pharmacological testing of compounds within the invention.
EXAMPLE 1
4-Hydroxy-3-(1-oxodecyl)-2(5H)-furanone
To a solution of 300 mg (3.0 mmol) of tetronic acid and 629 mg (685 .mu.L, 3.3 mmol) of decanoyl chloride is added 527 .mu.L (4.5 mmol) of tin(IV) tetrachloride. The solution is heated in an oil bath at 100.degree. C. for 3 hours. The reaction mixture is worked up by dissolving the cooled black solution in dichloromethane and washing with 1.0N HCl followed by brine. The organic layer is dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow oil. Chromatography on a 2 mm Chromatotron plate eluting with 5% methanol/dichloromethane gives an oil which is crystallized from ethyl acetate to give 119 mg (16%) of product, m.p 71.degree.-74.degree. C.
Spectral data follows. IR (KBr) 1780 (C.dbd.O), 1755 (C.dbd.O) cm.sup.-1.
Ms (EI) 254 (M+), 236 (-H.sub.2 O).
Analysis Calc'd. for C.sub.14 H.sub.22 O.sub.4.0.25 H.sub.2 O: C, 64.99; H, 8.70; N, 0.00. Found: C, 64.95; H, 8.24; N, 0.18.
EXAMPLE 2
4-Hydroxy-3-(1-oxo-3-phenylpropyl)-2(5H)-furanone
To a solution of 330 mg (3.3 mmole) of tetronic acid, 495 .mu.L (3.63 mmol) of triethylamine, and 132 mg (1.09 mmol) of 4-dimethylaminopyridine in 17 mL of dichloromethane is added at 0.degree. C., 403 .mu.L (3.63 mmol) of hydrocinnamoyl chloride. The iced bath is removed and stirred for 18 hours. The reaction mixture is worked up by pouring into 1.0N HCl and extracting three times with dichloromethane. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light yellow solid which is recrystallized from ethyl acetate/hexane to give 363 mg (47%) of product.
Spectral data follows. .sup.1 H NMR (400 MHz, d.sub.6 -DMSO) .delta. 7.11-7.29 (m, 5H, arom), 4.40 (bs, 2H, CH.sub.2 OC.dbd.O), 2.99 (t, 2H, J=6.7 Hz, CH.sub.2 C.dbd.O), 2.77 (t, 2H, J=6.7 Hz, CH.sub.2 Ph); IR (KBr) 1775 (C.dbd.O), 1750 (C.dbd.O) cm.sup.-1 ; MS (EI) 232 (M+), 214 (--H.sub.2 O).
Analysis Calc'd. for C.sub.13 H.sub.12 O.sub.4.0.75 H.sub.2 O: C, 63.54; H, 5.50; N, 0.00. Found: C, 63.83; H, 4.90; N, 0.21
EXAMPLE 3
4-Hydroxy-3-[(phenylmethoxy)acetyl]-2(5H)-furanone
To a solution of 1.0 g (10 mmol) of tetronic acid in 40 mL of dry dichloromethane is added at 0.degree. C., 1.5 mL (11 mmol) of triethylamine and 400 mg (3.3 mmol) of 4-dimethylaminopyridine. Then 2.03 g (1.74 mL, 11 mmol) of benzyloxyacetyl chloride is added dropwise. After 10 minutes, the ice bath is removed. The reaction mixture is stirred overnight at room temperature and is worked up by pouring into 1.0N HCl and extracting three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow solid. The solid is recrystallized from ethyl acetate to give 650 mg (26%) of product, m.p. 154.degree.-155.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 7.30-7.42 (m, 5H, arom), 4.73 (s, 2H, OCH.sub.2 Ph), 4.72 (bs, 2H, CH.sub.2 OC.dbd.O), 4.67 (s, 2H, OCH.sub.2 C.dbd.O); IR (KBr) 1755 (C.dbd.O) cm.sup.-1 ; MS (EI) no M+ peak, 142 (M-OCH.sub.2 Ph).
Analysis Calc'd. for C.sub.13 H.sub.12 O.sub.5 : C, 62.90; H, 4.87; N, 0.00. Found: C, 62.44; H, 5.02; N, 0.43.
EXAMPLE 4
4-Hydroxy-3-[3-(1H-indol-3-yl)-1-oxopropyl]-2(5H)-furanone
To a solution of 1.0 g (10 mmol) of tetronic acid in 30 mL of dry dichloromethane is added at 0.degree. C., 1.5 mL (11 mmol) of triethylamine and 400 mg (3.3 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 2.27 g (12 mmol) of 3-(1H-indol-3-yl)propionic acid is added followed by 2.29 g (12 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a brown solid. Trituration with ether gave 2.28 g (84%) of product, m.p. 148.degree.-149.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, d.sub.6 -acetone) .delta. 10.02 (bs, 1H, NH), 7.65 (d, 1H, J=7.1 Hz, arom), 7.36 (d, 1H, J=7.1 Hz, arom), 7.20 (s, 1H, arom), 7.09 (t, 1H, J=7.1 Hz, arom), 7.02 (t, 1H, J=7.1 Hz, arom), 4.72 (bs, 2H, CH.sub.2 OC.dbd.O), 3.30 (t, 2H, J=6.5 Hz, CH.sub.2 C.dbd.O), 3.02 (t, 2H, J=6.5 Hz, CH.sub.2 Ar); IR (KBr) 1750 (C.dbd.O) cm.sup.-1 ; MS (EI) 271 (M+), 144, 130.
Analysis Calc'd. for C.sub.15 H.sub.13 NO.sub.4. 0.25 H.sub.2 O: C, 65.34; H, 4.90; N, 5.08. Found: C, 65.26; H, 4.89; N, 5.33.
EXAMPLE 5
4-Hydroxy-3-[4-(1H-indol-3-yl)-1-oxobutyl]-2(5H)-furanone
To a solution of 775 mg (7.75 mmol) of tetronic acid in 30 mL of dry dichloromethane is added at 0.degree. C., 1.16 mL (8.53 mmol) of triethylamine and 310 mg (2.58 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 1.89 g (9.3 mmol) of 4-(1H-indol-3-yl)butyric acid is added followed by 1.78 g (9.3 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a brown solid. Trituration with ether gives 1.51 g (68%) of product, m.p 133.degree.-135.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, d.sub.6 -acetone) .delta. 9.90 (bs, 1H, NH), 7.52 (d, 1H, J=7.3 Hz, arom), 7.32 (d, 1H, J=7.3 Hz, arom), 7.10 (s, 1H, arom), 6.95 (t, 1H, J=7.3 Hz, arom), 6.93 (t, 1H, J=7.3 Hz, arom), 4.55 (bs, 2H, CH.sub.2 OC.dbd.O), 2.95 (t, 2H, J=6.5 Hz, CH.sub.2 C.dbd.O), 2.85 (t, 2H, J=6.5 Hz, CH.sub.2 Ar), 2.08 (m, 2H, CH.sub.2); IR (KBr) 1760 (C.dbd.O) cm.sup.-1 ; MS (EI) 285 (M+), 130.
Analysis Calc'd. for C.sub.16 H.sub.15 NO.sub.4. 0.25 H.sub.2 O: C, 66.32; H, 5.35; N, 4.84. Found: C, 66.71; H, 5.73; N, 6.10.
EXAMPLE 6
(E)-4-Hydroxy-3-(3-(4-methoxyphenyl)-1-oxo-2-propenyl)-2 (5H)-furanone
Into a stirring solution of 750 mg (5.27 mmol) of 3-acetyl tetronic acid and 636 mg (4.53 mmol) of p-anisaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl is continued for 2 hours at room temperature. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol affording 125 mg (10%) of product.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 8.01 (d, 0.5H, J=15.9 Hz, olefin), 7.99 (d, 0.5H, J=15.9 Hz, olefin), 7.87 (d, 0.5H, J=25.9 Hz, olefin), 7.80 (d, 0.5H, J=25.9 Hz, olefin), 7.65 (d, 2H, J=8.4 Hz, arom), 7.44 (d, 2H, J=8.4 Hz, arom), 4.66 (s, 2H, OCH.sub.2 C.dbd.O), 3.97 (s, 3H, OCH.sub.3); IR (KBr) 3260, 1750 (C.dbd.O), 1720 (C.dbd.O), 1650, 1570, 1550 cm.sup.-1.
Analysis Calc'd. for C.sub.14 H.sub.12 O.sub.5 : C, 64.61; H, 4.61; N, 0.00. Found: C, 64.52; H, 4.42; N, 0.00.
EXAMPLE 7
(E)-4-Hydroxy-3-(3-(4-chlorophenyl)-1-oxo-2-propenyl)-2(5H)-furanone
Into a stirring solution of 750 mg (5.27 mmol) of 3-acetyl tetronic acid and 636 mg (4.53 mmol) of p-chlorobenzaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl gas is continued for 2 hours. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol affording 523 mg (44%) of product, m.p 158.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 8.00 (d, 0.5H, J=16.0 Hz, olefin) 7.98 (d, 0.5H, J=16.0 Hz, olefin), 7.77 (d, 0.5H, J=26.0 Hz, olefin), 7.70 (d, 0.5H, J=26.0 Hz, olefin), 7.64 (d, 2H, J=8.3 Hz, arom), 7.44 (d, 2H, J=8.3 Hz, arom), 4.67 (s, 1H, OCH.sub.2 C.dbd.O), 4.61 (s, 1H, OCH.sub.2 C.dbd.O), 3.10 (bs, 1H, OH); IR (KBr) 3160, 1750 (C.dbd.O), 1700 (C.dbd.O), 1640, 1580, 1570 cm.sup.-1 ; MS (EI) 266 (M+), 264 (100), 246, 165, 137.
Analysis Calc'd. for C.sub.13 H.sub.9 O.sub.4 Cl: C, 59.00; H, 3.34; N, 0.00. Found: C, 58.57; H, 3.57; N, 0.00.
EXAMPLE 8
(E)-4-Hydroxy-3-(3-(phenyl)-1-oxo-2-propenyl)-2(5H)-furanone
Into a stirring solution of 500 mg (3.52 mmol) of 3-acetyl tetronic acid and 308 .mu.L (3.03 mmol) of benzaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl is continued for 2 hours. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol affording 387 mg (56%) of product, m.p 136.degree.-138.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 8.07 (d, 0.5H,J=16.0 Hz, olefin) 8.05 (d, 0.5H, J=16.0 Hz, olefin), 7.81 (d, 0.5H, J=28.1 Hz, olefin), 7.77 (d, 0.5H, J=28.1 Hz, olefin), 7.71-7.72 (m, 2H, arom), 7.43-7.51 (m, 3H, arom), 4.67 (s, 1H, OCH.sub.2 C.dbd.O), 4.60 (s, 1H, OCH.sub.2 C.dbd.O); IR (KBr) 3400, 1765 (C.dbd.O), 1755 (C.dbd.O); 1650 (C.dbd.O), 1620, 1575, 1560 cm.sup.-1 ; MS (EI) 230 (M+), 149 (100).
Analysis Calc'd. for C.sub.13 H.sub.10 O.sub.4 : C, 67.82; H, 4.38; N, 0.00. Found: C, 67.49; H, 4.49; N, 0.00.
EXAMPLE 9
(Z)-4-Hydroxy-3-(1-oxo-9-octadecenyl)-2(5H)-furanone
To a solution of 1.48 g (14.75 mmol) of tetronic acid in 45 mL of dry dichloromethane is added at 0.degree. C., 2.22 mL (16.22 mmol) of triethylamine and 592 mg (4.92 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 5.0 g (17.7 mmol) of oleic acid is added followed by 3.38 g (17.7 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light yellow solid. Trituration with cold pentane gives 3.85 g (72%) of product, 46.degree.-48.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, d.sub.6 -DMSO) .delta. 5.30 (m, 2H, olefin), 4.53 (bs, 2H, CH.sub.2 OC.dbd.O), 2.63 (t, 2H, J=6.0 Hz, CH.sub.2 C.dbd.O), 1.96 (m, 4H, CH.sub.2 C.dbd.C), 1.55-1.05 (m, 22H, CH.sub.2), 0.83 (t, 3H, J=6.0 Hz, CH.sub.3); IR (KBr) 1740 (C.dbd.O); MS (CI) 365 (M+1), 143, 171.
Analysis Calc'd. for C.sub.22 H.sub.36 O.sub.4 : C, 72.53; H, 9.89; N, 0.00. Found: C, 72.99; H, 10.12; N, 0.06.
EXAMPLE 10
(E)-4-Hydroxy-3-(1-oxo-3-(3-(trifluoromethyl)phenyl)-2-propenyl)-2 (5H)-furanone
Into a stirring solution of 750 mg (5.27 mmol) of 3-acetyl tetronic acid and 606 .mu.L (4.53 mmol) of m-trifluoromethylbenzaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl is continued for 2 hours. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol affording 263 mg (20%) of product, m.p 171.degree.-175.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, d.sub.6 -DMSO) .delta. 8.02 (d, 1H, J=16.1 Hz, olefin), 7.95 (m, 2H, arom), 7.75 (m, 1H, arom) 7.66 (m, 1H, arom), 7.64 (d, 1H, J=16.1 Hz, olefin), 4.37 (s, 2H, OCH.sub.2 C.dbd.O); IR (KBr) 3400, 1730 (C.dbd.O), 1705 (C.dbd.O), 1690 (C.dbd.O), 1630, 1600, 1555 cm.sup.-1 ; MS (EI) 298 (M+), 271, 239, 238, 199 (100), 151, 115.
Analysis Calc'd. for C.sub.14 H.sub.9 F.sub.3 O.sub.4 : C, 56.42; H, 3.04; N, 0.00. Found: C, 56.08; H, 2.99; N, 0.00.
EXAMPLE 11
4-Hydroxy-3-(1-oxoeicosanoyl)-2(5H)-furanone
To a stirring suspension of 8.76 g (87.6 mmol) of tetronic acid in 200 mL of dichloromethane at 0.degree. C. is added 13.4 mL (96.4 mmol) of triethylamine and 3.5 g (31.0 mmol) of 4-dimethylaminopyridine. After allowing the reaction mixture to stir for 5 minutes, 17 g (96.4 mmol) of eicosanoic acid and 20 g (105.0 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are added. The reaction mixture is allowed to stir at room temperature for 48 hours. Then, 100 mL of water is added to the reaction and the layers are separated. The aqueous layer is extracted three times with 50 mL of dichloromethane. The aqueous layer is acidified with 100 mL of 1.0N HCl and extracted three times with 100 mL of dichloromethane. The combined organic layers are dried over anhydrous sodium sulfate and concentrated in vacuo. The residue is triturated in ethyl ether and filtered, affording 14.3 g (64 %) of product, m.p 91.degree.-92.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 4.63 (bs, 2H, OCH.sub.2 C.dbd.O), 2.92 (t, 2H, J=7.6 Hz, CH.sub.2 C.dbd.O), 1.70 (m, 2H, CH2CH.sub.2 C.dbd.O), 1.27-1.41 (m, 12H, CH.sub.2), 0.88 (t, 3H, J=6.9 Hz, CH.sub.2 CH.sub.3); IR (KBr) 3200, 2900, 2850, 1780 (C.dbd.O), 1750 (C.dbd.O), 1660, 1650, 1615 cm.sup.-1 ; MS (EI) 255 (M+), 254, 236, 155, 142 (100), 127.
Analysis Calc'd. for C.sub.14 H.sub.22 O.sub.4 : C, 66.12; H, 8.72; N, 0.00. Found: C, 66.26; H, 8.59; N, 0.00.
EXAMPLE 12
(E)-4-Hydroxy-3-(3-(3-nitrophenyl)-1-oxo-2-propenyl)-2 (5H)-furanone
Into a stirring solution of 750 mg (5.27 mmol) of 3-acetyl tetronic acid and 686 mg (4.53 mmol) of 3-nitrobenzaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl is continued for 2 hours. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol affording 210 mg (17%) of product, m.p 168.degree.-170.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 8.49 (m, 1H, arom), 8.33 (m, 1H, arom), 8.05 (m, 2H, arom), 7.89 (d, 0.5H, J=18.5 Hz, olefin), 7.85 (d, 0.5H, J=18.5 Hz, olefin), 7.67 (t, 1H, J=8.0, arom), 4.71 (s, 1H, OCH.sub.2 C.dbd.O), 4.64 (s, 1H, OCH.sub.2 C.dbd.O); IR (KBr) 3400, 1755 (C.dbd.O), 1700 (C.dbd.O), 1630, 1610, 1580, 1560, 1530 cm.sup.-1 ; MS (EI) 275 (M+), 176, 69 (100).
Analysis Calc'd. for C.sub.13 H.sub.9 NO.sub.6 : C, 56.73; H, 3.30; N, 5.09. Found: C, 56.28; H, 3.11; N, 4.96.
EXAMPLE 13
(E)-3-(3-(2,5-Dimethoxyphenyl)-1-oxo-2-propenyl)-4-hydroxy-2 (5H)-furanone
Into a stirring solution of 750 mg (5.27 mmol) of 3-acetyl tetronic acid and 753 mg (4.53 mmol) of 2,5-dimethoxybenzaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl is continued for 2 hours. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol, affording 60 mg (4.5%) of product, m.p. 169.degree.-171.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 8.04 (d, 0.5H, J=10.0 Hz, olefin), 8.00 (d, 0.5H, J.dbd.10.0 Hz, olefin), 7.67 (d, 0.5H, J.dbd.15.8 Hz, olefin), 7.58 (d, 0.5H, J=15.8 Hz, olefin), 7.32 (m, 1H, arom), 7.20 (m, 1H, arom), 6.94 (m, 1H, arom), 4.65 (s, 1H, OCH.sub.2 C.dbd.O), 4.59 (s, 1H, OCH.sub.2 C.dbd.O), 3.97 (s, 6H, OCH.sub.3); IR (KBr) 3450, 1765 (C.dbd.O), 1690 (C.dbd.O), 1625, 1580, 1560, 1550, 1510 cm.sup.-1 ; MS (EI) 290 (M+), 137, 69 (100).
Analysis Calc'd. for C.sub.15 H.sub.14 O.sub.6 : C, 62.07; H, 4.86; N, 0.00. Found: C, 62.00; H, 4.87; N, 0.00.
EXAMPLE 14
(Z,Z)-4-Hydroxy-3-(1-oxo-9,12-octadecadienyl)-2(5H)-furanone
To a solution of 1.48 g (14.75 mmol) of tetronic acid in 45 mL of dry dichloromethane is added at 0.degree. C., 2.22 mL (16.22 mmol) of triethylamine and 592 mg (4.92 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 4.96 g (17.7 mmol) of linoleic acid is added, followed by 3.38 g (17.7 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0 N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light waxy yellow solid. Trituration with cold pentane gives 4.54 g (85%) of product.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 5.40 (m, 4H, olefin), 4.64 (bs, 2H, CH.sub.2 OC.dbd.O), 2.93 (t, 2H, J.dbd.6.4 Hz, CH.sub.2 C.dbd.O), 2.78 (m, 2H, C.dbd.CCH.sub.2 C.dbd.C), 2.06 (m, 4H, CH.sub.2 C.dbd.C), 1.71-1.36 (m, 16H, CH.sub.2), 0.92 (t, 3H, J.dbd.6.5 Hz, CH.sub.3); IR (KBr) 1775 (C.dbd.O) cm.sup.-1 ; MS (FAB) 363 (M+H).
Analysis Calc'd. for C.sub.22 H.sub.34 O.sub.4.0.25 H.sub.2 O: C, 72.03; H, 9.41; N, 0.00. Found: C, 71.93; H, 9.65; N, 0.00.
EXAMPLE 15
(Z,Z,Z)-4-Hydroxy-3-(1-oxo-9,12,15-octadecatrienyl)-2(5H)-furanone
To a solution of 1.48 g (14.75 mmol) of tetronic acid in 45 mL of dry dichloromethane is added at 0.degree. C., 2.22 mL (16.22 mmol) of triethylamine and 592 mg (4.92 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 4.92 g (17.7 mmol) of linolenic acid is added, followed by 3.38 g (17.7 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light brown waxy solid. Trituration with cold pentane gives 4.38 g (83%) of product.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 5.39 (m, 6H, olefin), 4.61 (bs, 2H, CH.sub.2 OC.dbd.O), 2.90 (t, 2H, J=7.0 Hz, CH.sub.2 C.dbd.O), 2.79 (m, 4H, C.dbd.CCH.sub.2 C.dbd.C), 2.08 (m, 4H, CH.sub.2 C.dbd.C), 1.68-1.34 (m, 10H, CH.sub.2), 0.99 (t, 3H, J=6.5 Hz, CH.sub.3); IR (KBr) 1775 (C.dbd.O) cm.sup.-1 ; MS (FAB) 361 (M+H).
Analysis Calc'd. for C.sub.22 H.sub.34 O.sub.4. H.sub.2 O: C, 69.84; H, 8.99; N, 0.00. Found: C, 69.68; H, 8.72; N 0.00.
EXAMPLE 16
(E)-3-(3-(3,4-Dimethoxyphenyl)-1-oxo-2-propenyl)-4-hydroxy-2 (5H)-furanone
Into a stirring solution of 750 mg (5.27 mmol) of 3-acetyl tetronic acid and 753 mg (4.53 mmol) of 3,4-dimethoxybenzaldehyde in 60 mL of methanol at -10.degree. C. is bubbled HCl gas for 3 hours. The ice bath is removed and HCl is continued for 2 hours. The reaction is concentrated in vacuo and the residue is recrystallized from ethanol affording 135 mg (10%) of product, m.p. 191.degree.-192.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 7.83 (d, 1H, J=15.9 Hz, olefin), 7.64 (d, 1H, J=15.9 Hz, olefin), 7.34 (d of d, 1H, J.sub.1 =1.9 Hz, J.sub.2 =8.5 Hz, arom), 7.28 (d, 1H, J=1.9 Hz, arom), 7.07 (d, 1H, J=8.5 Hz, arom), 4.59 (s, 2H, OCH.sub.2 C.dbd.O), 3.81 (s, 3H, OCH.sub.3); IR (KBr) 3450, 2940, 2840, 1760 (C.dbd.O), 1685 (C.dbd.O), 1630, 1580, 1555, 1505 cm.sup.-1 ; MS (EI) 291 (M+), 202, 85, 81, 71, 69 (100), 67.
Analysis Calc'd. for C.sub.15 H.sub.14 O.sub.6 : C, 62.07; H, 4.86; N, 0.00. Found: C, 61.70; H, 4.87; N, 0.00.
EXAMPLE 17
(Z)-4-Hydroxy-5,5-dimethyl-3-(1-oxo-9-octadecenyl)-2(5H)-furanone
To a solution of 1.89 g (14.75 mmol) of 5,5-dimethyltetronic acid in 45 mL of dry dimethylformamide is added at 0.degree. C., 2.22 mL (16.22 mmol) of triethylamine and 592 mg (4.92 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 5.00 g (17.7 mmol) of oleic acid is added followed by 3.38 g (17.7 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0 N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light yellow oil. Trituration with cold pentane gives 4.38 g (83%) of product.
Spectral data follows. .sup.1 H NMR (400 MHz, d.sub.6 -DMSO) .delta. 5.30 (m, 2H, olefin), 2.69 (t, 2H, J=6.5 Hz, CH.sub.2 C.dbd.O), 1.97 (m, 4H, CH.sub.2 C.dbd.C), 1.43-1.22 (m, 22H, CH.sub.2), 0.83 (t, 3H, J=6.5 Hz, CH.sub.3); IR (KBr) 1770 (C.dbd.O) cm.sup.-1 ; MS (EI) 392 (M+), 374, 183.
Analysis Calc'd. for C.sub.22 H.sub.34 O.sub.4.1.5 H.sub.2 O: C, 68.74; H, 10.26; N, 0.00. Found: C, 68.90; H, 9.44; N, 0.24.
EXAMPLE 18
(Z)-4-Hydroxy-3-(1-oxo-6-octadecenyl)-2(5H)-furanone
To a solution of 1.48 g (14.75 mmol) of tetronic acid in 45 mL of dry dimethylformamide is added at 0.degree. C., 2.22 mL (16.22 mmol) of triethylamine and 592 mg (4.92 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 5.00 g (17.7 mmol) of petroselenic acid is added, followed by 3.38 g (17.7 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light yellow solid. Recrystallization from cold pentane gives 3.20 g (28%) of product, m.p 41.degree.-42.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 5.35 (m, 2H, olefin), 4.58 (bs, 2H, CH.sub.2 OC.dbd.O), 2.72 (t, 2H, J=6.5 Hz, CH.sub.2 C.dbd.O), 1.98 (m, 4H, CH.sub.2 C.dbd.C), 1.58-1.20 (m, 22H, CH.sub.2), 0.84 (t, 3H, J=6.5 Hz, CH.sub.3); IR (KBr) 1775 (C.dbd.O), 1750 (C.dbd.O) cm.sup.-1 ; MS (EI) 364 (M+).
Analysis Calc'd. for C.sub.22 H.sub.36 O.sub.4 : C, 72.49; H, 9.95; N, 0.00. Found: C, 72.05; H, 9.93; N, 0.02.
EXAMPLE 19
(Z)-4-Hydroxy-3-(1-oxo-8-(2'-octylcyclopropyl)octanyl)-2(5H)-furanone
To a solution of 1.48 g (14.75 mmol) of tetronic acid in 45 mL of dry dimethylformamide is added at 0.degree. C., 2.22 mL (16.22 mmol) of triethylamine and 592 mg (4.92 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 5.25 g (17.7 mmol) of cis-8-(2'-octylcyclopropyl)octanoic acid is added followed by 3.38 g (17.7 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a colorless solid. Recrystallization from cold pentane gave 3.20 g (57%) of product, m.p 49.degree.-50.degree. C.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 4.62 (bs, 2H, CH.sub.2 OC.dbd.O), 2.95 (t, 2H, J=6.0 Hz, CH.sub.2 C.dbd.O), 1.71 (m, 2H, CH.sub.2), 1.45-1.20 (m, 22H, CH.sub.2), 1.15 (m, 2H, cyclopropyl CH.sub.2), 0.92 (t, 3H, J=6.5 Hz, CH.sub.3), 0.90-0.70 (m, 1.5H, cyclopropyl CH.sub.2), -0.35 (m, 0.5H, cyclopropyl CH.sub.2); IR (KBr) 1775 (C.dbd.O), 1760 (C.dbd.O) cm.sup.-1.
Analysis Calc'd. for C.sub.23 H.sub.38 O.sub.4 : C, 72.98; H, 10.12; N, 0.00. Found: C, 72.52; H, 9.94; N, 0.05.
EXAMPLE 20
(Z,Z,Z)-4-Hydroxy-3-(1-oxo-6,9,12-octadecatrienyl)-2(5H)-furanone
To a solution of 300 g (2.98 mmol) of tetronic acid in 10 mL of dry dimethylformamide is added at 0.degree. C., 450 .mu.L (3.29 mmol) of triethylamine and 121 mg (1.0 mmol) of 4-dimethylaminopyridine. After stirring for 5 minutes, 1.00 g (3.59 mmol) of gamma-linolenic acid is added followed by 668 mg (3.59 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After 10 minutes, the ice bath is removed. The reaction mixture is stirred at room temperature overnight. The reaction mixture is poured into 1.0N HCl and extracted three times with ethyl acetate. The organic layers are combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a brown oil. Flash chromatography on a 40 mm.times.150 mm silica column eluting with 5% methanol-ethyl acetate gives 150 mg (14%) of product.
Spectral data follows. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 5.41 (m, 6H, olefin), 4.62 (bs, 2H, CH.sub.2 OC.dbd.O), 2.94 (t, 2H, J=7.0 Hz, CH.sub.2 C.dbd.O), 2.80 (m, 4H, C.dbd.CCH.sub.2 C.dbd.C), 2.09 (m, 4CH.sub.2 C.dbd.C), 1.75-1.24 (m, 10H, CH.sub.2), 0.90 (t, 3H, J=6.5 Hz, CH.sub.3); IR (KBr) 1770 (C.dbd.O) cm.sup.-1 ; MS (EI) 360 (M+).
EXAMPLE 21
4-Hydroxy-3-[6-(4-chlorophenoxy)-1-oxohexyl]-2(5H)-furanone
To a stirring solution of 554 mg (5.54 mmol) of tetronic acid in 20 mL of diemthylformamide is added 850 .mu.L (6.09 mmol) of triethylamine and 220 mg (1.765 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 1.26 g (6.63 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.475 g (6.63 mmol) of 6-[4-(hexyloxy)phenoxy]hexanoic acid are added and the reaction mixture is stirred 2 days at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo giving a yellow solid. Pure material is obtained by trituration in ether, filtering to give 774 mg (43%) of 4-hydroxy-3-[6-(4-chlorophenoxy)-1-oxohexyl]-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) 7.216 (d, 2H, Ar), 6.818 (d, 2H, Ar), 4.681 (s, 2H, CH.sub.2 OC.dbd.O), 3.930 (t, 2H, CH.sub.2 OAr), 2.954 (t, 2H, O.dbd.CCH.sub.2), 1.836-1.559 (m, 6H, CH.sub.2); IR (KBr) 3200, 2980, 2920, 2860, 1775, 1750, 1660, 1620, 1490, 1245, 1050 cm.sup.-1 ; MS (EI) 324 (M+), 197, 142, 128 (100), 127.
Analysis Calc'd. for C.sub.16 H.sub.17 O.sub.5 Cl.0.25H.sub.2 O: C, 58.36; H, 5.36. Found: C, 58.78; H, 5.20.
EXAMPLE 22
4-Hydroxy-3-[1-oxo-8-[2-[(2-pentylcyclopropyl)methyl]cyclopropyl]octyl]-2(5H)-furanone
To a stirring solution of 947 mg (9.5 mmol) of tetronic acid in 30 mL of dimethylformamide is added 1.42 mL (10.45 mmol) of triethylamine and 379 mg (3.14 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 2.17 g (11.4 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 3.5 g (11.4 mmol) of (cis)-7-(2-octylcyclopropyl)octanoic acid are added and the reaction mixture is stirred 2 days at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are dried over Na.sub.2 SO.sub.4 and concentrated in vacuo giving a yellow oil. Pure material is obtained by flash chromatography on a 40 mm.times.150 mm silica column eluting with 50% ethyl acetate/hexane to 100% ethyl acetate giving 870 mg (20%) of 4-hydroxy-3-[1-oxo-8-[2-[(2-pentylcyclopropyl)methyl]cyclopropyl]octyl]- 2(5H)-furanone as an oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3,400 MHz) .delta. 4.572 (s, 2H, CH.sub.2 OC.dbd.O), 2.704 (t, 2H, O.dbd.CCH.sub.2), 1.481 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.341-1.111 (m, 20H, CH.sub.2), 0.845 (t, 3H, CH.sub.2 CH.sub.3), 0.747-0.644 (m, 4H, cyclopropyl), 0.559 (m, 2H, cyclopropyl), -0.332 (m, 2H, cyclopropyl); IR (KBr) 3060, 2990, 2910, 2840, 1770, 1695, 1655, 1600, 1455, 1435, 1195, 1040 cm.sup.-1 ; MS (EI) 390 (M+), 127 (100), 67(94).
Analysis Calc'd. for C.sub.24 H.sub.38 O.sub.4 : C, 73.81; H, 9.81. Found: C, 73.59; H, 9.83.
EXAMPLE 23
4-Hydroxy-3-((Z)-1-oxo-10-tetradecenyl)-2(5H)-furanone
To a stirring solution of 523 mg (5.22 mmol) of tetronic acid in 17 mL of dimethylformamide is added 800 .mu.L (5.75 mmol) of triethylamine and 210 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 1.19 g (5.75 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.2 g (5.75 mmol) of (Z)-10-tetradecenoic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted with four 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material is obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 1.2 g (75%) of 4-hydroxy-3-((Z)-1-oxo-10-tetradecyl)-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (d.sub.6 -DMSO, 400 MHz) .delta. 5.356 (m, 2H, HC.dbd.CH), 4.509 (s, 2H, CH.sub.2 OC.dbd.O), 2.686 (t, 2H, O.dbd.CCH.sub.2), 1.932 (m, 4H, CH.sub.2 C.dbd.CCH.sub.2), 1.470 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.225 (s, 12H, CH.sub.2), 0.851 (t, 3H,CH.sub.2 CH.sub.3); IR (KBr) 2900, 2820, 1770, 1745, 1690, 1650, 1610, 1455, 1435, 1385, 1340, 1305, 1275, 1235, 1230, 1125,1060, 1015 cm.sup.-1 ; MS (EI) 310, 308 (M.sup.+.), 290,155, 142 (100), 127.
Analysis Calc'd. for C.sub.18 H.sub.28 O.sub.4 : C, 70.10; H, 9.15. Found: C, 69.89; H, 9.43.
EXAMPLE 24
(Z)-9-Octadecanoic acid 5-(2,5-dihydro-4-hydroxy-3-furanyl)-5-oxo-1,2-pentanediyl ester
A. 4,5-Bis[((Z)-1-oxo-9-octadecenyl)oxy]pentanoic acid
To a stirring solution of 3.76 g (16.1 mmol) of (Z)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-1-pentene-1,2-diol in 50 mL of methylene chloride is added 4.9 mL (35.2 mmol) of triethylamine and 1.3 g of 4-dimethylaminopyridine at 0.degree. C. Next, 7.4 g (38.6 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 10 g (35.4 mmol) of oleic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is poured into water and extracted with four 50 mL portions of methylene chloride. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo. The resulting crude diester dissolved in 32 mL of tetrahydrofuran and treated with 32 mL (32 mmol) of 1.0 N tetrabutylammonium fluoride in tetrahydrofuran and the reaction is stirred 2 hours at room temperature. The reaction is poured into water and extracted with four 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material was obtained by flash chromatography with 15% ethyl acetate/hexane as eluant giving 7.7 g (77%) of alcohol. Next, 20 mL of 2.0M Jones reagent is added at 0.degree. C. to a stirring solution of 7.7 g (11.86 mmol) of the alcohol and the reaction is stirred for 2 hours. The reaction is poured into water and extracted with four 50 ml portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material was obtained by flash chromatography with 35% ethyl acetate/hexane as eluant giving 2.2 g (28%) of 4,5-bis[((Z)-1-oxo-9-octadecyl)oxy]pentanoic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.344 (m, 4H, HC.dbd.CH), 5.175 (m, 1H, HCOC.dbd.O), 4.145 (m, 2H, H.sub.2 COC.dbd.O), 2.422 (t, 2H, HO.sub.2 CCH.sub.2), 2.301 (t, 4H, O(C.dbd.O)CH.sub.2), 2.012 (m, 8H, C.dbd.CCH.sub.2), 1.612 (m, 4H, O(C.dbd.O)CH.sub.2 CH.sub.2), 1.289 (m, 40H, CH.sub.2), 0.881 (t, 6H, CH.sub.2 CH.sub.3); IR (KBr) 3000, 2920, 2840, 1740,1710,1460, 1160 cm.sup.-1 ; MS (CI.sup.+) 664 (MH.sup.++), 663 (MH.sup.+), 645, 381 (100), 338, 265, 159, 87.
Analysis Calc'd. for C.sub.41 H.sub.74 O.sub.6 : C, 74.27; H, 11.25. Found: C, 73.92; H, 11.28.
B) (Z)-9-Octadecanoic acid 5-(2,5-dihydro-4-hydroxy-3-furanyl)-5-oxo-1,2-pentanediyl ester
To a stirring solution of 137 mg (1.37 mmol) of tetronic acid in 10 mL of dimethylformamide are added 220 .mu.L (1.5 mmol) of triethylamine and 70 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 330 mg (1.72 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.0 g (1.5 mmol) of 4,5-bis[((Z)-1-oxo-9-octadecenyl)oxy]pentanoic acid and the reaction is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted with four 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material was obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 608 mg (60%) of (Z)-9-octadecanoic acid 5-(2,5-dihydro-4-hydroxy-3-furanyl)-5-oxo-1,2-pentanediyl ester.
Spectral data follows: .sup.1 H NMR (d.sub.6 -DMSO, 400 MHz) .delta. 6.68 (bs, 1H, OH), 5.290 (m, 4H, HC.dbd.CH), 4.980 (s, 1H, HCOC.dbd.O), 4.500 (s, 2H, CH.sub.2 OC.dbd.O), 4.216 (s, 1H, CH.sub.2 OC.dbd.O), 3.974 (s, 1H, CH.sub.2 OC.dbd.O), 2.726 (m, 2H, O.dbd.CCH.sub.2), 2.218 (s, 4H, O.dbd.CCH.sub.2), 1.953 (m, 8H, C.dbd.CCH.sub.2), 1.477 (s, 4H, O.dbd.CCH.sub.2 CH.sub.2), 1.220 (s, 40H, CH.sub.2), 0.830 (t, 6H, CH.sub.2 CH.sub.3); IR (KBr) 3000, 2900, 2840, 1760,1730, 1690, 1650, 1600, 1460, 1450, 1430, 1220, 1165, 1035, 1010 cm.sup.-1 ; MS (CI.sup.+) 463, 409 (100), 265, 69.
Analysis Calc'd. for C.sub.45 H.sub.76 O.sub.8 : C, 72.54; H, 10.28. Found: C, 72.58; H, 10.48.
EXAMPLE 25
4-Hydroxy-3-((Z)-1-oxo-9-tetradecenyl)-2(5H)-furanone
To a stirring solution of 111 mg (1.11 mmol) of tetronic acid in 5 mL of dimethylformamide is added 185 .mu.L (1.33 mmol) of triethylamine and 100 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 303 mg (1.33 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 300 mg (1.33 mmol) of myristoleic acid are added and the reaction mixture is stirred 3 days at room temperature. The reaction is acidified with 1.0N HCl and extracted with four 100 mL portions of ethyl acetate. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material is obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 280 mg (82%) of 4-hydroxy-3-((Z)-1-oxo-9-tetradecenyl)-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.349 (m, 2H, HC.dbd.CH), 4.615 (s, 2H, CH.sub.2 OC.dbd.O), 2.919 (t, 2H, O.dbd.CCH.sub.2), 2.010 (m, 4H, CH.sub.2 C.dbd.CCH.sub.2), 1.704 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.314 (m, 12H, CH.sub.2), 0.896 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3200, 3000, 2930, 2860, 1775, 1750, 1660, 1615,1960, 1435, 1390, 1345, 1275,1235,1125,1060,1015 cm.sup.-1 ; MS (EI) 308 (M.sup.+.), 290, 155, 142 (100), 127, 69, 67.
Analysis Calc'd. for C.sub.18 H.sub.28 O.sub.4 : C, 70.10; H, 9.15. Found: C, 70.01; H, 9.50.
EXAMPLE 26
4-Hydroxy-3-((Z)-1-oxo-9-hexadecenyl)-2(5H)-furanone
To a stirring solution of 1.07 g (10.72 mmol) of tetronic acid in 30 mL of dimethylformamide is added 1.64 mL (11.97 mmol) of triethylamine and 428 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 2.44 g (12.77 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 3.0 g (11.79 mmol) of palmitoleic acid are added and the reaction is stirred for 3 days at room temperature. The reaction is acidified with 1.0N HCl and extracted with four 100 mL portions of ethyl acetate. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material was obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 1.3 g (36%) of 4-hydroxy-3-((Z)-1-oxo-9-hexadecenyl)-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.343 (m, 2H, HC.dbd.CH), 4.615 (d, 2H, CH.sub.2 OC.dbd.O), 2.918 (t, 2H, O.dbd.CCH.sub.2), 2.019 (m, 4H, C.dbd.CCH.sub.2), 1.704 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.312 (m, 16H, CH.sub.2), 0.883 (t, 3H, CH.sub.2 CH.sub.3), IR (KBr) 3200, 3000, 2920, 2850, 1775,1750, 1660, 1615, 1460, 1440, 1390, 1345, 1305, 1275, 1235, 1230, 1125, 1060, 1015 cm.sup.-1 ; MS (EI) 337 (MH.sup.+), 336 (M.sup.+.), 318, 155, 142 (100), 127, 69.
Analysis Calc'd. for C.sub.20 H.sub.32 O.sub.4 : C, 71.39; H, 9.59. Found: C, 70.92; H, 5.96.
EXAMPLE 27
4-Hydroxy-3-(1-oxo-9-octadecynyl)-2(5H)-furanone
To a stirring solution of 1.62 g (17.86 mmol) of tetronic acid in 50 mL of dimethylformamide is added 2.5 mL (17.86 mmol) of triethylamine and 652 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 3.71 g (19.35 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 5.0 g (17.86 mmol) of stearolic acid are added and the reaction is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted with three 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material was obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 3.3 g (56%) of 4-hydroxy-3-(1-oxo-9-octadecynyl)-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.616 (s, 2H, CH.sub.2 OC.dbd.O), 2.930 (t, 2H, O.dbd.CCH.sub.2), 2.137 (m, 4H, C.dbd.CCH.sub.2), 1.710 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.380 (m, 20H, CH.sub.2), 0.881 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3100, 2960, 2935, 2850, 1755, 1695, 1665, 1600, 1570, 1465, 1430, 1390, 1290, 1175, 1035, 1020, 865 cm.sup.-1 ; MS (EI) 362 (M.sup.+.), 344, 260 (100), 246, 155, 142, 127, 95, 81, 67.
Analysis Calc'd. for C.sub.22 H.sub.34 O.sub.4 : C, 72.89; H, 9.45. Found: C, 73.23; H, 9.63.
EXAMPLE 28
4-Hydroxy-3-(1-oxooctadecyl)-2(5H)-furanone
To a stirring solution of 1.6 g (16 mmol) of tetronic acid in 75 mL of methylene chloride is added 2.5 mL (17.86 mmol) of triethylamine and 640 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 5.0 g (17.6 mmol) of stearoyl chloride is added and the reaction is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted with two 100 mL portions of methylene chloride. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material was obtained by trituration with ethyl acetate as affording 3.3 g (58%) of 4-hydroxy-3-(1-oxooctadecyl)-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.615 (s, 2H, CH.sub.2 OC.dbd.O), 2.912 (t, 2H, O.dbd.CCH.sub.2), 1.684 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.388-1.111 (m, 28H, CH.sub.2), 0.881 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3450, 2960, 2920, 2850, 1760, 1650, 1610, 1030, 885 cm.sup.-1 ; MS (EI) 366 (M.sup.+.), 348, 155, 142(100), 127.
Analysis Calc'd. for C.sub.22 H.sub.38 O.sub.4 : C, 72.09; H, 10.45. Found: C, 72.42; H, 10.60.
EXAMPLE 29
3-[9-(4-Chlorophenoxy)-1-oxononyl]-4-hydroxy-2(5H)-furanone
To a stirring solution of 736 mg (7.36 mmol) of tetronic acid in 30 mL of dimethylformamide is added 1.2 mL (8.09 mmol) of triethylamine and 300 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 1.7 g (8.86 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.6 g (8.06 mmol) of 9-(4-chlorophenoxy)nonanoic acid are added and the reaction is stirred overnight at room temperature. The reaction is acidified with 1.0 N HCl and extracted with four 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material is obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 800 mg (30%) of 3-[9-(4-chlorophenoxy)-1-oxononyl]4-hydroxy-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.216 (d, 2H, Ar), 6.813 (d, 2H, Ar), 4.614 (s, 2H, CH.sub.2 OC.dbd.O), 3.911 (t, 2H, OCH.sub.2 Ar), 2.912 (t, 2H, O.dbd.CCH.sub.2), 1.742 (m, 4H, CH.sub.2), 1.370 (m, 8H, CH.sub.2); IR (KBr) 3220, 2940, 2860, 1770, 1745, 1660, 1610, 1600, 1495, 1475, 1435, 1395, 1345, 1290, 1270, 1250, 1210, 1175, 1130, 1110, 1100, 1060, 1010, 825 cm.sup.-1 ; MS (CI.sup.+) 367 (MH.sup.+), 277, 275 (100), 257, 239, 91, 61.
Analysis Calc'd for C.sub.19 H.sub.23 ClO.sub.5 : C, 62.21; H, 6.32. Found: C, 61.82; H, 6.25.
EXAMPLE 30
Hexylcarbamic acid 5-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-5-oxo-1,2-pentanediyl ester
To a stirring solution of 234 mg (2.34 mmol) of tetronic acid in 20 mL of dimethylformamide is added 400 .mu.L (2.57 mmol) of triethylamine and 200 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 600 mg (3.12 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 1.0 g (2.57 mmol) of 4,5-bis[[(hexylamino)carbonyl]-oxy]pentanoic acid are added and the reaction is stirred 3 days at room temperature. The reaction is acidified with 1.0N HCl and extracted with three 100 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording 636 mg (58%) of hexylcarbamic acid 5-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-5-oxo-1,2-pentanediyl ester.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.896 (m, 1H, HCOC.dbd.O), 4.792 (m, 2H, NH), 4.628 (s, 2H, CH.sub.2 OC.dbd.O), 4.157 (m, 2H, H.sub.2 COC.dbd.O), 3.152 (m, 6H, NHCH.sub.2, O.dbd.CCH.sub.2), 2.047 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.485 (m, 4H, HNCH.sub.2 CH.sub.2), 1.289 (m, 12H, CH.sub.2), 0.885 (t, 6H, CH.sub.2 CH.sub.3); IR (KBr) 3350, 2960, 2935, 2860, 1750, 1700, 1645, 1580, 1550, 1465, 1275, 1040 cm.sup.-1 ; MS (pos.ion FAB) 515 (MNa.sup.++), 493 (MNa.sup.+), 370, 350, 181, 102, 55, 43 (100).
Analysis Calc'd. for C.sub.23 H.sub.38 N.sub.2 O.sub.8 : C, 58.71; H, 8.14; N, 5.95. Found: C, 58.77; H, 8.19; N, 5.77.
EXAMPLE 31
Dodecylcarbamic acid 5-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-5-oxo-1,2-pentanediyl ester
A) 4,5-Bis[[(Dodecylamino)carbonyl]oxy]pentanoic acid
To a stirring solution 1.9 g (9.47 mmol) of 1.9 g (9.47 mmol) of the (Z)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-1-pentene-1,2-diol in 20 mL of methylene chloride is added 3.2 mL (23 mmol) of triethylamine and 4.8 g (9.47 mmol) of dodecyl isocyanate at 0.degree. C. and the reaction is stirred for 2 days at room temperature. The reaction is diluted with methylene chloride and washed with 1.0N HCl, saturated sodium bicarbonate, and brine, dried over sodium sulfate and concentrated in vacuo. The resulting dicarbamate is dissolved in 20 mL of tetrahydrofuran and treated with 20 mL of 1.0M tetrabutylammonium fluoride in tetrahydrofuran and the reaction is stirred overnight at room temperature. The reaction mixture was poured into water and extracted with three 100 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material is obtained by flash chromatography with ethyl acetate as eluant affording 4.1 g (80%) of alcohol. Next, 12 mL of 2.0M Jones reagent is added at 0.degree. C. to a stirring solution of 4.1 g (7.62 mmol) of the alcohol in 75 mL of acetone and the reaction is stirred for 4 hours. The reaction is poured into water and extracted with three 100 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo, affording 2.1 g (50%) of 4,5-bis[[dodecylamino)carbonyl]oxy]pentanoic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.961 (m, 1H, HCOC.dbd.O), 4.741 (m, 2H, NH), 4.110 (m, 2H, H.sub.2 COC.dbd.O), 3.130 (m, 4H, HNCH.sub.2), 2.425 (m, 2H, HO.sub.2 CCH.sub.2), 1.897 (m, 2H, HO.sub.2 CCH.sub.2 CH.sub.2), 1.466 (m, 4H, HNCH.sub.2 CH.sub.2), 1.242 (s, 3H, CH.sub.2), 0.866 (t, 6H, CH.sub.2 CH.sub.3); IR (KBr) 3450, 2930, 2860, 1700, 1660, 1470, 1280 cm.sup.-1 ; MS (pos.ion FAB) 579 (M.sup.+ Na), 557 (MH.sup.+), 328 (100), 230, 186, 117, 91, 73, 57, 43, 30.
Analysis Calc'd. for C.sub.31 H.sub.60 N.sub.2 O.sub.6 : C, 66.87; H, 10.86; N, 5.03, Found: C, 67.30; H, 11.06; N, 5.06.
B) Dodecylcarbamic acid 5-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-5-oxo-1,2-pentanediyl ester
To a stirring solution of 163 mg (1.63 mmol) of tetronic acid in 20 mL of dimethylformamide is added 250 .mu.L (1.8 mmol) of triethylamine and 100 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 400 mg (2.08 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.0 g (1.8 mmol) of 4,5-bis[[(dodecylamino)carbonyl]oxy]pentanoic acid are added and the reaction is stirred 3 days at room temperature. The reaction is acidified with 1.0N HCl and extracted with three 100 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude compound. Pure material is obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 500 mg (50%) of dodecylcarbamic acid 5-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-5-oxo-1,2-pentanediyl ester.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.896 (m, 1H, HCOC.dbd.O), 4.792 (m, 2H, NH), 4.628 (s, 2H, CH.sub.2 OC.dbd.O), 4.157 (m, 2H, H.sub.2 COC.dbd.O), 3.152 (m, 6H, O.dbd.CCH.sub.2, NHCH.sub.2), 2.047 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.485 (m, 4H, NHCH.sub.2 CH.sub.2), 1.289 (m, 36H, CH.sub.2), 0.885 (t, 6H, CH.sub.2 CH.sub.3); IR (KBr) 3350, 2960, 2935, 2860, 1750, 1700, 1645, 1580, 1550, 1465, 1275, 1040 cm.sup.-1 ; MS (pos.ion FAB) 683 (MNa.sup.++), 661 (MNa.sup.+), 237, 221,186,131,91,73,57 (100).
Analysis Calc'd. for C.sub.35 H.sub.62 N.sub.2 O.sub.8 : C, 65.80; H, 9.78; N, 4.38, Found: C, 66.19; H, 9.86; N, 4.26.
EXAMPLE 32
4-Hydroxy-3-[6-[4-[(4-chlorophenyl)methoxyphenoxy]]1-oxohexyl]-2(5H)-furanone
To a stirring solution of 757 mg (7.57 mmol) of tetronic acid in 50 mL of dimethylformamide is added 1.20 mL (8.83 mmol) of triethylamine and 400 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 2.00 g (8.3 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.88 g (8.3 mmol) of 6-[4-(4-chlorophenyl)methoxyphenoxy]hexanoic acid are added and the reaction mixture is stirred 3 days at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo giving a yellow oil. Pure material was obtained by flash chromatography on a 40 mm.times.150 mm silica column eluting with 10% methanol/ethyl acetate giving 800 mg (25%) of 4-hydroxy-3-[6-[4-[(4-chlorophenyl)methoxyphenoxy]]-1-oxohexyl]-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.338 (s, 4H, Ar), 6.849 (d, 2H, Ar), 6.811 (d, 2H, Ar), 4.966 (s, 2H, CH.sub.2 Ar), 4.611 (s, 2H, CH.sub.2 OC.dbd.O), 3.899 (t, 2H, CH.sub.2 OAr), 2.964 (t, 2H, O.dbd.CCH.sub.2), 1.824-1.546 (m, 6H, CH.sub.2); IR (KBr) 3420, 2945, 2860, 1770, 1660, 1615, 1515, 1245, 1030 cm.sup.-1 ; MS (EI) 430 (M+), 197, 127, 125 (100).
Analysis Calc'd. for C.sub.23 H.sub.23 O.sub.6 Cl: C, 64.11; H, 5.38. Found: C, 63.98; H, 5.45.
EXAMPLE 33
4-Hydroxy-3-[6-[4-(hexyloxy)phenoxy]-1-oxohexyl]-2(5H)-furanone
To a stirring solution of 876 mg (8.76 mmol) of tetronic acid in 50 mL of dimethylformamide is added 1.40 mL (10.30 mmol) of triethylamine and 400 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 2.00 g (8.3 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.97 g (8.3 mmol) of 6-[4-(hexyloxy)phenoxy]hexanoic acid are added and the reaction mixture is stirred 3 days at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo giving a yellow oil. Pure material was obtained by flash chromatography on a 40 mm.times.150 mm silica column eluting with 10% methanol/ethyl acetate giving 1.1 g (32%) of 4-hydroxy-3-[6-[4-(hexyloxy)phenoxy]-1-oxohexyl]-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.601 (s, 2H, CH.sub.2 OC.dbd.O), 3.910 (s, 4H, CH.sub.2 OAr), 2.951 (t, 2H, O.dbd.CCH.sub.2), 1.815-1.309 (m, 14H, CH.sub.2), 0.918 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 2950, 2880, 1770, 1680, 1650, 1615, 1520, 1250, 1040 cm.sup.-1 ; MS (CI) 391 (MH+), 349, 291, 117 (100).
Analysis Calc'd for C.sub.22 H.sub.30 O.sub.6 : C, 67.67; H, 7.74. Found: C, 67.70; H, 7.70.
EXAMPLE 34
4-Hydroxy-3-[7-(4-chlorophenoxy)-1-oxoheptyl]-2(5H)-furanone
To a stirring solution of 1.00 g (10 mmol) of tetronic acid in 40 mL of dimethylformamide is added 1.51 mL (11.11 mmol) of triethylamine and 402 mg (3.33 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 2.29 g (11.97 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 3.07 g (11.97 mmol) of 7-(4-chlorophenoxy)heptanoic acid are added and the reaction mixture is stirred 2 days at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are dried over Na.sub.2 SO.sub.4 and concentrated in vacuo giving a yellow oil. Pure material was obtained by flash chromatography on a 40 mm.times.150 mm silica column eluting with 10% methanol/ethyl acetate giving 1.50 g (44%) of 4-hydroxy-3-[7-(4-chlorophenoxy)-1-oxoheptyl]-2(5H)furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3,400 MHz) .delta. 7.290 (d, 2H, Ar), 6.928 (d, 2H, Ar), 4.488 (s, 2H, CH.sub.2 OC.dbd.O), 3.924 (t, 2H, CH.sub.2 OAr), 2.711 (t, 2H, O.dbd.CCH.sub.2), 1.694-1.298 (m, 8H,CH.sub.2); IR (KBr) 3210, 2960, 2880, 1773, 1760, 1665, 1620, 1500, 1480, 1440, 1355, 1060 cm.sup.-1 ; MS (EI) 338 (M+), 211 (100), 193, 128.
Analysis Calc'd. for C.sub.17 H.sub.19 O.sub.5 Cl.0.25 H.sub.2 O: C, 59.48; H, 5.69, Found: C, 59.27; H, 5.79.
EXAMPLE 35
3-[(Z)-10-(4-chlorophenyl)-1-oxo-9-decenyl]-4-hydroxy-2(5H)-furanone
A) (Z)-10-(4-chlorophenyl)-9-decenoic acid
To a stirring solution of 20.0 g (79.7 mmol) of methyl-9-bromononate in 50 mL of acetonitrile is added 21.0 g (79.7 mmol) of triphenylphosphine and allowed to reflux overnight. The reaction mixture is concentrated in vacuo affording 41.0 g (100%) of phosphonium salt. Next, 8.5 g (16.5 mmol) of the phosphonium salt is dissolved in 100 mL of tetrahydrofuran and the reaction is cooled to -78.degree. C. 17.0 mL (17.0 mmol) of 1.0M LiHMDS is added dropwise and the reaction mixture is stirred for 1 hour. Then, 2.3 g (16.5 mmol) of 4-chlorobenzaldehyde is added and the reaction mixture is stirred overnight with gradual warming to room temperature. The reaction mixture is poured into water and extracted with three 30 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material was obtained by flash chromatography with 10% ethyl acetate/hexane as eluant affording 600 mg (12%) of olefin. Next, 2.0 g of KOH is added at room temperature to a solution of 600 mg (1.93 mmol) of olefin in a 3:1 solution of methanol/water and the reaction mixture is stirred for 2 hours. The reaction mixture is acidified with 1.0N HCl and extracted with three 50 mL portions of ether. The organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording 500 mg (87%) of (Z)-10-(4-chlorophenyl)-9-decenoic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.287 (d, 2H, Ar), 7.186 (d, 2H, Ar), 6.343 (d, 1H, HC.dbd.CH), 5.665 (m, 1H, HC.dbd.CH), 2.301 (m, 4H, HOCCH.sub.2, C.dbd.CCH.sub.2), 1.618 (m, 2H, HO.sub.2 CCH.sub.2 CH.sub.2), 1.433 (m, 2H, C.dbd.CCH.sub.2 CH.sub.2), 1.304 (m, 6H, CH.sub.2).
B) 3-[(Z)-10-(4-chlorophenyl)-1-oxo-9-decenyl]-4-hydroxy-2(5H)-furanone
To a stirring solution of 117.0 mg (117.0 mmol) of tetronic acid in 10 mL of dimethylformamide is added 181 .mu.L (1.3 mmol) of triethylamine and 50 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 264 mg (1.37 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 380 mg of (Z)-10-(4-chlorophenyl)-9-decenoic acid are added and the reaction mixture is stirred 2 days at room temperature. The reaction mixture is acidified with 1.0N HCl and extracted with three 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material was obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 100 mg (25%) of 3-[(Z)-10-(4-chlorophenyl)-1-oxo-9-decynyl]-4-hydroxy-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.221 (d, 2H, Ar), 7.115 (d, 2H, Ar), 6.279 (d, 1H, HC.dbd.CH), 5.596 (d of t, 1H, HC.dbd.CH), 4.544 (s, 2H, CH.sub.2 OC.dbd.O), 2.828 (t, 2H, O.dbd.CCH.sub.2), 2.202 (m, 2H, C.dbd.CCH.sub.2), 1.617 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.266 (m, 8H, CH.sub.2); IR (KBr) 3450, 3200, 2950, 2880, 1785, 1735, 1675, 1620, 1500, 1475, 1450, 1400, 1360, 1270, 1240, 1110, 1070, 1025, 885 cm.sup.-1 ; MS (EI) 364 (MH.sup.+), 362 (MH.sup.+), 328, 327, 309, 279, 167, 151, 149 (100), 138, 127, 95, 71.
Analysis Calc'd. for C.sub.20 H.sub.23 ClO.sub.4 : C, 66.20; H, 6.39. Found : C, 66.58; H, 6.43.
EXAMPLE 36
4-Hydroxy-3-((E)-1-oxo-9-octadecenyl)-2(5H)-furanone
To a stirring solution of 1.0 g (10.0 mmol) of tetronic acid in 32 mL of dimethylformamide is added 1.53 mL (10.98 mmol) of triethylamine and 400 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 2.28 g (11.98 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 3.1 g (10.98 mmol) of elaidic acid are added and the reaction mixture is stirred for 2 days at room temperature. The reaction mixture is acidified with 1.0N HCl and extracted with four 100 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo. Trituration of the crude product in ether affords 1.2 g (33%) of 4-hydroxy-3-((E)-1-oxo-9-octadecynyl)-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.367 (m, 2H, HC.dbd.CH), 4.602 (s, 2H, CH.sub.2 OC.dbd.O), 2.897 (t, 2H, O.dbd.CCH.sub.2), 1.943 (m, 4H, C.dbd.CCH.sub.2), 1.689 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.314 (m, 20H, CH.sub.2), 0.867 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3420, 2920, 2840, 1770, 1750, 1660, 1615, 1455, 1430, 1385, 1340, 1265, 1235, 1220, 1200, 1120, 1050, 1010, 960, 940, 830 cm.sup.-1 ; MS (EI) 364 (M.sup.+.), 346, 155, 142 (100), 127.
Analysis Calc'd. for C.sub.22 H.sub.36 O.sub.4 : C, 72.49; H, 9.95. Found: C, 72.50; H, 9.89.
EXAMPLE 37
3-[6-(4-Heptylphenoxy)-1-oxohexyl]-4-hydroxy-2(5H)-furanone
To a stirring solution of 1.1 g (10.98 mmol) of tetronic acid in 25 mL of dimethylformamide is added 1.53 mL (10.98 mmol) of triethylamine and 400 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 2.28 g (11.98 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 3.38 g (10.98 mmol) of 6-(4-heptylphenoxy)heptanoic acid are added and the reaction mixture is stirred for 3 days at room temperature. The reaction mixture is acidified with 1.0N HCl and extracted with three 200 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo. Trituration of the crude product in ethyl acetate/hexane affords 2.1 g (50%) of 3-[6-(4-heptylphenoxy)-1-oxohexyl]-4-hydroxy-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.071 (d, 2H, Ar), 6.797 (d, 2H, Ar), 4.608 (s, 2H,CH.sub.2 OC.dbd.O), 3.935 (t, 2H, OCH.sub.2), 2.949 (t, 2H, O.dbd.CCH.sub.2), 2.523 (t, 2H, CH.sub.2 Ar), 1.786 (m, 4H, O.dbd.CCH.sub.2 CH.sub.2), 1.573 (m, 4H, ArCH.sub.2 CH.sub.2), 1.284 (m, 8H, CH.sub.2), 0.873 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3450, 3200, 2920, 2850, 1780, 1750, 1660, 1615, 1510, 1460, 1430, 1385, 1340, 1245, 1175, 1125, 1050, 1010, 830 cm.sup.-1 ; MS (EI) 388 (M.sup.+.), 197, 192, 127, 107 (100).
Analysis Calc'd. for C.sub.23 H.sub.32 O.sub.5 : C, 71.10; H, 8.30. Found: C, 71.23; H, 8.36.
EXAMPLE 38
(Z)-9-Octadecanoic acid 4-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-4-oxobutanoic ester
A) (Z)-9-octadecenoic acid 4-hydroxybutyl ester
To a stirring solution of 2.0 g (7.0 mmol) of oleic acid and 1.6 g (7.0 mmol) of 2-(4-bromobutoxy)tetrahydro-2H-pyran in 20 mL of tetrahydrofuran is added 2.3 g (7.1 mmol) of cesium carbonate and the reaction mixture is stirred overnight at room temperature. The reaction mixture is poured into water and extracted with three 100 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording 2.9 g (100%) of (Z)-9-octadecenoic acid 4-[(tetrahydro-2H-pyran-2-yl)oxy]butyl ester. Next, 500 mg of Dowex 50W 8X resin is added to a stirring solution of 2.9 g (7.0 mmol) of (Z)-9-octadecenoic acid 4-[(tetrahydro-2H-pyran-2-yl)oxy]butyl ester in 100 mL of 1:1 tetrahydrofuran/methanol and the reaction mixture is stirred overnight at room temperature. The reaction mixture is filtered and concentrated in vacuo affording 2.4 g (100%) of alcohol. Next, 11 mL of 2.0M Jones reagent is added at 0.degree. C. to a stirring solution of 2.4 g (7.0 mmol) of alcohol in 75 mL of acetone and the reaction mixture is stirred for 2 hours. The reaction mixture is poured into water and extracted with three 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording 2.1 g (81%) of title compound.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.332 (m, 2H, HC.dbd.CH), 4.126 (t, 2H, O.dbd.COCH.sub.2), 2.405 (t, 2H, HO.sub.2 CCH.sub.2), 2.283 (t, 2H, O(C.dbd.O)CH.sub.2), 2.003 (m, 4H, C.dbd.CCH.sub.2), 1.595 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.270 (m, 22H, CH.sub.2), 0.867 (t, 3H, CH.sub.2 CH.sub.3).
B) (Z)-9-Octadecanoic acid 4-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-4-oxobutanoic ester
To a stirring solution of 570 mg (5.7 mmol) of tetronic acid in 50 mL of dimethylformamide is added 795 .mu.L (5.7 mmol) of triethylamine and 400 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 1.2 g (6.26 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.1 g of (Z)-9-octadecenoic acid 4-hydroxybutyl ester are added and the reaction mixture is stirred overnight at room temperature. The reaction mixture is acidified with 1.0N HCl and extracted with three 100 mL portions of ethyl acetate. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material is obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 1.8 g (72%) of (Z)-9-octadecenoic acid 4-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-4-oxobutanoic ester.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.332 (m, 2H, HC.dbd.CH), 4.616 (s, 2H, CH.sub.2 OC.dbd.O), 4.126 (t, 2H, O.dbd.COCH.sub.2), 3.004 (t, 2H, O.dbd.CCH.sub.2), 2.283 (t, 2H, O(C.dbd.O)CH.sub.2), 2.003 (m, 4H, C.dbd.CCH.sub.2), 1.595 (m, 2H, O.dbd.CCH.sub.2 CH.sub.2), 1.270 (m, 22H, CH.sub.2), 0.867 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3025 2935, 2865, 1775, 1735, 1705, 1610, 1520, 1470, 1440, 1230, 1180, 1045, 1015 cm.sup.-1 ; MS (CI.sup.+) 451 (MH.sup.+), 297, 283, 265, 183, 169 (100), 102.
Analysis Calc'd. for C.sub.26 H.sub.42 O.sub.6 : C, 69.30; H, 9.39. Found: C, 69.08; H, 9.37.
EXAMPLE 39
4-Hydroxy-3-[6-(1,1,3,3-tetramethylbutyl)phenoxy)-1-oxohexyl]2(5H)-furanone
To a stirring solution of 329 mg (3.29 mmol) of tetronic acid in 15 mL of dimethylformamide is added 458 .mu.L (3.29 mmol) of triethylamine and 220 mg (1.765 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 681 mg (3.58 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrocholoride and 1.0 g (3.29 mmol) of 6-(1,1,3,3-tetramethybutyl)phenoxy)hexanoic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo giving a yellow solid. Flash chromatography on a 40 mm.times.150 mm silica gel column eluting with 10% methanol/ethyl acetate gives 650 mg (50%) of 4-hydroxy-3-[6-(1,1,3,3-tetramethylbutyl)phenoxy)-1-oxohexyl]-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.255 (d, 2H, Ar), 6.778 (d, 2H, Ar), 4.599 (s, 2H, CH.sub.2 OC.dbd.0), 3.935 (t, CH.sub.2 OAr), 2.947 (t, 2H, O.dbd.CCH.sub.2), 1.824-1.573 (m, 6H, CH.sub.2), 1.688 (s, 2H, CH.sub.2), 1.330 (3, 6H, C(CH.sub.3).sub.2), 0.705 (s, 9H, t-Bu); IR (KBr) 2960, 2870, 1775, 1695, 1655, 1610, 1510, 1250, 1040 cm.sup.1 ; MS (EI) 402 (M+), 331 (100), 127.
Analysis Calc'd. for C.sub.24 H.sub.34 O.sub.5 : C, 71.61; H, 8.51. Found: C, 71.70; H, 8.42.
EXAMPLE 40
((Z)-9-octadecynyl)carbamic acid 1-[[((E)-3,7-Dimethyl-2,6-octadecadienyl)oxy]methyl]-4-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-4-oxobutyl ester
A) 5-[((E)-3,7-Dimethyl-2,6-octadienyl)oxy]-4-[[((Z)-9-octadecenylamino)carbonyl]oxy]pentanoic acid
To a stirring suspension of 140 mg (4.6 mmol) of 80% NaH in 5 mL of tetrahydrofuran is added 1.07 g (4.6 mmol) of (Z)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-1-pentene-1,2-diol at 0.degree. C. and the reaction mixture is stirred for 1 hour. Next, 1.0 g (4.6 mmol) of geranyl bromide is added and the reaction is stirred overnight at room temperature. The reaction mixture is poured into water and extracted with three 50 mL portions of ethyl acetate. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording 1.7 g (100%) of (E)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-1-[(3,7-dimethyl-2,6-octadienyl)oxy]-2-pentanol. Next, 1.6 g (4.3 mmol) of (E)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-1-[(3,7-dimethyl-2,6-octadienyl)oxy]-2-pentanol and 800 .mu.L (5.73 mmol) of triethylamine in 20 mL of methylene chloride are added at 0.degree. C. to a stirring solution of 13.3 mL of 20% phosgene in toluene and the reaction mixture is stirred for 1 hour. The reaction mixture is concentrated in vacuo and redissolved in 20 mL of tetrahydrofuran. Next, 1.3 g (4.6 mmol) of oleyl amine is added and the reaction mixture is stirred for 2 hours. The reaction mixture is poured into water and extracted with three 50 mL portions of ether. The organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material is obtained by flash chromatography with 10% ethyl acetate/hexane as eluant affording 1.67 g (67%) of (Z)-9-octadecenylcarbamic acid (E)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-1-[[(3,7-dimethyl-2,6-octadienyl)oxy]methyl]butyl ester. Next, 5 mL of 1.0M tetrabutylammonium fluoride in tetrahydrofuran is added to a stirring solution of 1.67 g (2.5 mmol) of (Z)-9-octadecenylcarbamic acid (E)-5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]- 1-[[(3,7-dimethyl-2,6-octadienyl)oxy]methyl]butyl ester in 5 mL of tetrahydrofuran and the reaction mixture is stirred overnight. The reaction mixture is poured into water and extracted with three 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material is obtained by flash chromatography with 10% ethyl acetate/hexane as eluant affording 722 mg (54%) of (Z)-9-octadecenylcarbamic acid (E)-1-[[(3,7-dimethyl-2,6-octadienyl)oxy]methyl]-4-hydroxybutyl ester. Next, 2 mL of 2.0M Jones reagent is added at 0.degree. C. to a solution of 722 mg (1.33 mmol) of (Z)-9-octadecenylcarbamic acid (E)-1-[[(3,7-dimethyl-2,6-octadienyl)oxy]methyl]-4-hydroxybutyl ester in 12 mL of acetone and the reaction is stirred for 3 hours. The reaction is poured into water and extracted with three 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material was obtained by flash chromatography with 35% ethyl acetate/hexane as eluant affording 226 mg (30%) of 5-[((E)-3,7-dimethyl-2,6-octadienyl)oxy]-4-[[((Z)-9-octadecenylamino) carbonyl]oxy]pentanoic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.336 (m, 3H, HC.dbd.CH), 5.076 (m, 1H, HC.dbd.C), 4.903 (m, 1H, HCOC.dbd.O), 4.751 (m, 1H, NH), 4.001 (m, 2H, OCH.sub.2), 3.487 (d, 2H, OCH.sub.2), 3.143 (d, 2H, O.dbd.CCH.sub.2, HNCH.sub.2), 2.422 (t, 2H, HO.sub.2 CCH.sub.2), 2.076 (m, 8H, C.dbd.CCH.sub.2), 1.666 (s, 3H, C.dbd.CCH.sub.3), 1.640 (s, 3H, C.dbd.CCH.sub.3), 1.588 (s, 3H, C.dbd.CCH.sub.3), 1.466 (m, 4H, O.dbd.CCH.sub.2 CH.sub.2, NHCH.sub.2 CH.sub.2), 1.256 (m, 22H, CH.sub.2), 0.869 (t, 3H, CH.sub.2 CH.sub.3); IR (KBr) 3350, 2940, 2870, 1725, 1545, 1455, 1380, 1250, 1130 cm.sup.-1 ; MS (CI.sup. +) 564 (MH.sup.+), 429, 428, 410, 153, 137 (100), 135, 117, 99, 81.
Analysis Calc'd. for C.sub.34 H.sub.61 NO.sub.5 : C, 72.42; H, 10.90; N, 2.48. Found: C, 72.13; H, 10.53; N, 2.41.
B) ((Z)-9-octadecynyl)carbamic acid 1-[[((E)-3,7-Dimethyl-2,6-octadecadienyl)oxy]methyl]-4-(2,5-dihydro-4-hydroxy-2-oxo-3-furanyl)-4-oxobutyl ester
To a stirring solution of 146 mg (1.46 mmol) of tetronic acid in 10 mL of dimethylformamide is added 200 .mu.L (1.46 mmol) of triethylamine and 75 mg of 4-dimethylaminopyridine at 0.degree. C. Next, 300 mg (1.56 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 810 mg (1.46 mmol) of 5-[((E)-3,7-dimethyl-2,6-octadienyl)oxy]-4-[[((Z)-9-octadecenylamino)carbonyl]oxy]pentanoic acid are added and the reaction mixture is stirred for 2 days at room temperature. The reaction mixture is acidified with 1.0N HCl and extracted with three 50 mL portions of ether. The combined organic layers are dried over MgSO.sub.4 and concentrated in vacuo affording crude product. Pure material was obtained by flash chromatography with 10% methanol/ethyl acetate as eluant affording 800 mg (86%) of ((Z)-9-octadecynyl)carbamic acid 1-[[((E)-3,7-dimethyl-2,6-octadecadienyl)oxy]methyl]-4-(2,5-dihydro-4-hydroxy-2-oxo-3 -furanyl)-4-oxobutyl ester.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.351 (m, 3H, HC.dbd.CH), 5.090 (m, 1H, HC.dbd.C), 4.918 (m, 1H, HCOC.dbd.O), 4.747 (m, 1H, NH), 4.667 (s, 2H, CH.sub.2 OC.dbd.O), 4.039 (m, 2H, OCH.sub.2), 3.515 (d, 2H, OCH.sub.2), 3.136 (m, 2H, NHCH.sub.2), 3.033 (m, 2H, O.dbd.CCH.sub.2), 2.076 (m 8H, C.dbd.CCH.sub.2), 1.683 (s, 3H, C.dbd.CCH.sub.3), 1.658 (s, 3H, C.dbd.CCH.sub.3), 1.605 (s, 3H, C.dbd.CCH.sub.3), 1.483 (m, 4H, O.dbd.CCH.sub.2 CH.sub.2, HNCH.sub.2 CH.sub.2), 1.256 (m, 22H, CH.sub.2), 0.865 (t, 3H, CH.sub.2 CH.sub.3); IR (film) 3360, 2930, 2860, 1775, 1725, 1705, 1605, 1525, 1460, 1440, 1375, 1245, 1125, 1040, 1015 cm.sup.-1 ; MS (CI.sup.+) 646 (MH.sup.+), 511, 510, 492, 412, 411, 410 (100), 209, 199, 153, 137, 81.
Analysis Calc'd. for C.sub.38 H.sub.63 NO.sub.7 : C, 70.66; H, 9.83; N, 2.17. Found: C, 70.60; H, 9.48; N, 1.92.
EXAMPLE 41
4-Hydroxy-3-[10-(4-chlorophenyl)-1-oxodecyl]-2(5H)-furanone
To a solution of 165 mL (165 mmol) of a 1.0M solution of boranetetrahydrofuran complex in tetrahydrofuran in 200 mL of dry diethyl ether at 0.degree. C. is added dropwise 25.0 g (123 mmol) of azelaic acid monomethyl ester, added at such a rate as to prevent excessive release of gas and exothermicity. The solution is allowed to warm to room temperature overnight and is worked up by slowly adding water until gas evolution ceases. Then solid potassium carbonate is added and the organic layer is separated. The aqueous layer is extracted three times with diethyl ether. The combined extracts are washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo to give 8-carbomethoxyoctan-1-ol as a colorless oil.
This crude oil is dissolved in 800 mL of dry dichloromethane and 417 mg of 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical is added followed by 4.25 g of sodium bromide in 42 mL of water. The solution is cooled to 0.degree.-10.degree. C. and 417 mL of a 5.25% solution of NaOCl (Clorox) and 8.34 g of sodium bicarbonate are added dropwise to the stirring solution. The solution is stirred an additional hour after addition was complete. The organic layer is separated, washed with aqueous sodium thiosulfate, followed by a saturated sodium bicarbonate solution and then by brine. The solution is dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo to give a 19.0 g (83%) of 8-carbomethoxyoctan-1-al as a light yellow oil.
To a solution of 18.2 g (43 mmol) of 4-chlorobenzyltriphenylphosphonium chloride in 200 mL of dry tetrahydrofuran at -78.degree. C. is added dropwise, 30.7 mL (43 mmol) of a 1.4M solution of potassium hexamethyldisilazide in tetrahydrofuran. This is allowed to stir for 1 hour when 8.0 g (43 mmol) of 8-carbomethoxyoctanal is added dropwise. The solution is stirred at -78.degree. C. for 2 hours, allowed to warm to room temperature, and stirred for 2 days. The reaction is worked up by adding water and extracting three times with diethyl ether. The ether extracts are combined, washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo to give a yellow solid. Pad filtration through flash silica gel eluting with 10% ethyl acetate/hexane, gives a yellow oil after concentration in vacuo. Flash chromatography on a 60 mm.times.150 mm silica column eluting with 5% ethyl acetate/hexane gives 1.62 g (13%) of 10-(4-chlorophenyl)-dec-9-enoic acid methyl ester as a mixture of E and Z isomers.
To a solution of 1.62 g of 10-(4-chlorophenyl)-dec-9-enoic acid methyl ester in 200 mL of 4:1 methanol:water is added 5.4 g of KOH. The solution is stirred for 2 hours and extracted three times with diethyl ether. The aqueous layer is acidified with concentrated hydrochloric acid, and extracted three times with diethyl ether. The organic layers are combined, washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo to give a yellow oil. The residue is purified via flash chromatography on a 40 mm.times.150 mm silica column eluting with 40% ethyl acetate/hexane to give 1.6 g (100%) of pure 10-(4-chlorophenyl)dec-9-enoic acid as a mixture of E and Z isomers.
To a stirring solution of 189 mg (1.89 mmol) of tetronic acid in 15 mL of dimethylformamide is added 290 .mu.L (2.08 mmol) of triethylamine and 8 mg (66 .mu.mol) of 4-dimethylaminopyridine at 0.degree. C. Next, 427 mg (2.22 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 690 mg (2.20 mmol) of 10-(4-chlorophenyl)dec-9-enoic acid as a mixture of E and Z isomers are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over Na.sub.2 SO.sub.4, and concentrated in vacuo. The residue is purified via flash chromatography on a 40 mm.times.150 mm silica gel column eluting with 5% methanol/ethyl acetate to give 400 mg (58%) of pure 4-hydroxy-3-[10-(4-chlorophenyl)-1-oxodec-9-enyl] -2(5H)-furanone as a mixture of E and Z isomers.
To a solution of 1.58 g (4.35 mmol) of 4-hydroxy-3-[10-(4-chlorophenyl)-1-oxodec-9-enyl]-2(5H)-furanone in 100 mL of ethyl acetate is added 250 mg of 5% palladium on carbon and subjected to atmospheric hydrogenation using a balloon to deliver the hydrogen. After 1.5 hours, the reaction is worked up by filtering through Solka Flok. The filtrate is washed with copious amounts of ethyl acetate and concentrated in vacuo to give 1.05 g (66%) of pure 4-hydroxy-3-[10-(4-chlorophenyl)1-oxodecyl]-2(5H)-furanone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta.7.22 (d, 2H, arom), 7.09 (d, 2H, arom), 4.66 (s, 2H, CH.sub.2 OC.dbd.O), 2.90 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.56 (t, 2H, CH.sub.2 Ar), 1.69 (m, 2H, CH.sub.2), 1.57 (m, 2H, CH.sub.2), 1.42-1.22 (m, 10H, CH.sub.2); IR (KBr) 2920, 2840, 1760, (C.dbd.O), 1745, 1655, 1610 cm.sup.-1 ; MS (EI) 364 (M+), 329, 155, 142, 125 (100).
Analysis Calc'd. for C.sub.20 H.sub.25 ClO.sub.4 : C, 65.84; H, 6.91. Found: C, 66.03; H, 7.02.
EXAMPLE 42
4-Hydroxy-3-[10-(3,4-dichlorophenyl)-1-oxodecyl]-2(5H)-furanone
Using the procedure of Example 41 for the synthesis of 4-hydroxy-3-[10-(4-chlorophenyl)-1-oxodecyl]-2(5H)-furanone above, but using 3,4-dichlorobenzyltriphenylphosphonium bromide in place of 4-chlorobenzyltriphenylphosphonium chloride, there is obtained the title compound.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.29 (m, 2H, arom), 6.97 (m, 1H, arom), 4.64 (s, 2H, CH.sub.2 OC.dbd.O), 2.90 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.25 (t, 2H, CH.sub.2 Ar), 1.67 (m, 2H, CH.sub.2), 1.54 (m, 2H, CH.sub.2), 1.42-1.22 (m, 10H, CH.sub.2); IR (KBr) 3180, 2920, 2845, 1760 (C.dbd.O), 1655, 1610 cm.sup.-1 ; MS (EI) 398 (M+), 159, 127 (100).
Analysis Calc'd. for C.sub.20 H.sub.24 Cl.sub.2 O.sub.4 : C, 60.16; H, 6.06. Found: C, 60.55; H, 5.65.
EXAMPLE 43
4-Hydroxy-3-[10-[4-(1,1-dimethylethyl)phenyl]1-oxodecyl]-2(5H)-furanone
Using the procedure of Example 41, but using 4-(1,1-dimethylethyl)benzyltriphenylphosphonium bromide in place of 4-chlorobenzyltriphenylphosphonium chloride, there is obtained the title compound.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 7.29 (d, 2H, arom), 7.11 (d, 2H, arom), 4.65 (s, 2H, CH.sub.2 OC.dbd.O), 2.92 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.57 (t, 2H, CH.sub.2 Ar), 1.70 (m, 2H, CH.sub.2), 1.59 (m, 2H, CH.sub.2), 1.42-1.22 (m, 10H, CH.sub.2); IR (KBr) 3410, 2910, 2845, 1775 (C.dbd.O), 1695, 1650, 1605 cm.sup.-1 ; MS (EI) 386 (M+), 371, 147, 127 (100).
Analysis Calc'd. for C.sub.24 H.sub.34 O.sub.4 : C, 74.58; H, 8.87. Found: C, 73.95; H, 8.67.
EXAMPLE 44
4-Hydroxy-3-[10-(4-bromophenyl)-1-oxodecyl]-2(5H)-furanone
Using the procedure of Example 41, but using 4-bromobenzyltriphenylphosphonium bromide in place of 4-chlorobenzyltriphenylphosphonium chloride, there is obtained the title compound.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3 , 400 MNz) .delta. 7.37 (d, 2H, arom), 7.04 (d, 2H, arom), 4.64 (s, 2H, CH.sub.2 OC.dbd.O), 2.91 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.54 (t, 2H, CH.sub.2 Ar), 1.69 (m, 2H, CH.sub.2), 1.59 (m, 2H, CH.sub.2), 1.42-1.22 (m, 10H, CH.sub.2); IR (KBr) 3200, 2930, 2850, 1765 (C.dbd.O), 1660, 1610 cm.sup.-1 ; MS (CI+) 409 (MH+).
Analysis Calc'd. for C.sub.20 H.sub.25 BrO.sub.4 : C, 58.69; H, 6.16. Found: C, 59.24; H, 6.12.
EXAMPLE 45
4-Hydroxy-3[10-(4-trifluoromethyl)phenyl]-2(5H)-furanone
Using the procedure of Example 41, but using 4-trifluoromethylbenzyltriphenylphosphonium bromide in place of 4-chlorobenzyltriphenylphosphonium chloride, there is obtained the title compound.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3,400 MHz) .delta. 7.52 (d, 2H, arom), 7.26 (d, 2H, arom), 4.63 (s, 2H, CH.sub.2 OC.dbd.O), 2.91 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.65 (t, 2H, CH.sub.2 Ar), 1.70 (m, 2H, CH.sub.2), 1.61 (m, 2H, CH.sub.2), 1.42-1.22 (m, 10H, CH.sub.2); IR (KBr) 3400, 2920, 2840, 1770 (C.dbd.O), 1695, 1650, 1605 cm.sup.-1 ; MS (neg. FAB) 397 (M-H).
Analysis Calc'd. for C.sub.21 H.sub.25 F.sub.3 O.sub.4 : C, 63.31; H, 6.32. Found: C, 62.97; H, 6.48.
EXAMPLE 46
4-Hydroxy-3[(Z,Z)-1-oxo-9,12-octadecadienyl]-2(5H)-thiophenone
To a stirring solution of 1.71 g (14.75 mmol) of thiotetronic acid in 10 mL of dimethylformamide is added 2.22 ML (17.2 mmol) of triethylamine and 592 mg (4.85 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 3.38 g (17.6 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4.96 g (17.7 mmol) of linoleic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with diethyl ether. The combined organic layers are washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo. The residue is recrystallized from pentane at -78.degree. C. to give a solid which melts upon warming to room temperature giving 1.67 g (30%) of 4-hydroxy-3-[(Z,Z)-1-oxo-9,12-octadecadienyl]-2(5H)-thiophenone as an oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.35 (m, 4H, CH.sub.2 CH.dbd.CHCH.sub.2), 3.98 (s, 2H, CH.sub.2 SC.dbd.O), 2.95 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.77 (m, 2H, CH.dbd.CHCH.sub.2 CH.dbd.CH), 2.02 (m, 4H, CH.sub.2 CH.dbd.CH), 1.65 (m, 2H, CH.sub.2), 1.42-1.22 (m, 14H, CH.sub.2), 0.88 (t, 3H, J=7.5 Hz, CH.sub.3); IR (Film) 2995, 2905, 2840, 1683 (C.dbd.O), 1612, 1565 cm.sup.-1 ; MS (CI+) 379 (MH+), 281 (100), 263.
Analysis Calc'd. for C.sub.22 H.sub.34 SO.sub.3 : C, 69.80; H, 9.05. Found: C, 70.10; H, 9.28.
EXAMPLE 47
4-Hydroxy-3-[(Z)-1-oxo-9-octadecenyl]-2(5H)-thiophenone
To a stirring solution of 1.71 g (14.75 mmol) of thiotetronic acid in 10 mL of dimethylformamide is added 2.22 mL (17.2 mmol) of triethylamine and 592 mg (4.85 mmol) of 4-diemthylaminopyridine at 0.degree. C. Next, 3.38 g (17.6 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 5.0 g (17.7 mmol) of oleic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with diethyl ether. The combined organic layers are washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo. The residue is subjected to HPLC chromatography (2 inch Dynamax silica gel column, 25 mL/min) eluting with 100% ethyl acetate to give 1.85 g (33%) of 4-hydroxy-3-[(Z)-1-oxo-9-octadecenyl]-2(5H)-thiophenone as an oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.34 (m, 2H, CH.sub.2 CH.dbd.CHCH.sub.2), 3.98 (s, 2H, CH.sub.2 SC.dbd.O), 2.96 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.01 (m, 4H, CH.sub.2 CH.dbd.CH), 1.67 (m, 2H, CH.sub.2), 1.42-1.20 (m, 20H, CH.sub.2), 0.88 (t, 3H, J=7.5 Hz, CH.sub.3); IR (Film) 2990, 2905, 2840, 1685 (C.dbd.O), 1615, 1565 cm.sup.-1 ; MS (EI) 380 (M+), 362, 158 (100), 143, 116.
EXAMPLE 48
4-Hydroxy-3-[(Z)-1-oxo-9-tetra-decenyl]-2(5H)-thiophenone
To a stirring solution of 855 mg (7.37 mmol) of thiotetronic acid in 10 mL of dimethylformamide is added 902 .mu.L (6.47 mmol) of triethylamine and 240 mg (1.96 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 1.68 g (8.76 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.0 g (8.84 mmol) of myristoleic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0 N HCl and extracted three times with diethyl ether. The combined organic layers are washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo. The residue is subjected to HPLC chromatography (2 inch Dynamax silica gel column, 25 mL/min) eluting with 100% ethyl acetate to give 785 mg (33%) of 4-hydroxy-3-[(Z)-1-oxo-9-tetradecenyl]-2(5H)-thiophenone as an oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta.5.35 (m, 2H, CH.sub.2 CH.dbd.CHCH.sub.2), 3.98 (s, 2H, CH.sub.2 SC.dbd.O), 2.95 (t, 2H, J.dbd.7.5 Hz, O.dbd.CCH.sub.2), 2.02 (m, 4H, CH.sub.2 CH.dbd.CH), 1.66 (m, 2H, CH.sub.2), 1.31 (m, 12H, CH.sub.2), 0.90 (t,3H, J.dbd.7.5 Hz, CH.sub.3); IR (Film) 3020, 2920, 2860, 1695 (C.dbd.O), 1625, 1560 cm.sup.-1 ; MS (EI) 324 (M+), 306, 157, 69 (100).
EXAMPLE 49
4-Hydroxy-3-[5-(4-chlorophenoxy)-1-oxopentyl]-2(5H)-thiophenone
To a stirring solution of 224 mg (1.89 mmol) of thiotetronic acid in 10 mL of dimethylformamide is added 290 .mu.L (2.08 mmol) of triethylamine and 77 mg (631 .mu.mol) of 4-dimethylaminopyridine at 0.degree. C. Next, 440 mg (2.29 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 530 mg (2.32 mmol) of 5-[4-(chlorophenoxy]pentanoic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ether. The combined organic layers are dried over Na.sub.2 SO.sub.4 and concentrated in vacuo giving a yellow solid. The residue is recrystallized from ethyl acetate/hexane at -78.degree. C. to give 150 mg (24%) of 4-hydroxy-3-[5-(4-chlorophenoxy)-1-oxopentyl]-2(5H)-thiophenone.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta.7.20 (d, 2H, Ar), 6.79 (d, 2H, Ar), 3.95 (s, 2H, CH.sub.2 SC.dbd.O), 3.94 (t, 2H, CH.sub.2 OAr), 3.03 (t, 2H, O.dbd.CCH.sub.2), 1.85 (m, 4H, CH.sub.2); IR (KBr) 3410, 2960, 2940, 2880, 1680, 1625, 1580, 1490 cm.sup.-1 ; MS (EI) 326 (M+), 199, 143, 128 (100).
Analysis Calc'd. for C.sub.15 H.sub.15 SO.sub.4 Cl: C, 55.13; H, 4.63, Found: C, 55.46; H, 4.78.
EXAMPLE 50
4-Hydroxy-3-[(Z,Z,Z)-1-oxo-6,9,12-octadecatrienyl]-2(5H)-thiophenone
To a stirring solution of 695 mg (6.04 mmol) of thiotetronic acid in 10 mL of dimethylformamide is added 902 .mu.L (6.47 mmol) of triethylamine and 240 mg (1.97 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 1.37 g (7.13 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.0 g (7.19 mmol) of .gamma.-linolenic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with diethyl ether. The combined organic layers are washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo. The residue was subjected to HPLC chromatography (2 inch Dynamax silica gel column, 25 mL/min) eluting with 100% ethyl acetate to give 1.8 g (79%) of 4-hydroxy-3-[(Z,Z,Z)-1-oxo-6,9,12-octadecatrienyl]-2(5H)-thiophenone as an oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.38 (m, 6H, CH.sub.2 CH.dbd.CHCH.sub.2), 3.98 (s, 2H, CH.sub.2 SC.dbd.O), 2.97 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.81 (m, 4H, CH.dbd.CHCH.sub.2 CH.dbd.CH), 2.08 (m, 4H, CH.sub.2 CH.dbd.CH), 1.69 (m, 2H, CH.sub.2), 1.47 (m, 2H, CH.sub.2), 1.30 (m, 6H, CH.sub.2), 0.88 (t, 3H, J=7.5 Hz, CH.sub.3); IR (Film) 2980, 2900, 2820, 1695 (C.dbd.O), 1675, 1615, 1550 cm.sup.-1 ; MS (CI+) 377 (MH+).
EXAMPLE 51
4-Hydroxy-3-[(Z)-1-oxo-6-octadecenyl]-2(5H)-thiophenone
To a stirring solution of 1.71 g (14.7 mmol) of thiotetronic acid in 20 mL of dimethylformamide is added 2.22 mL (15.9 mmol) of triethylamine and 592 mg (4.85 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 3.38 g (17.6 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 5.0 g (17.7 mmol) of petroselinic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with diethyl ether. The combined organic layers are washed with brine, dried over Na.sub.2 SO.sub.4, filtered, and concentrated in vacuo. The residue is subjected to HPLC chromatography (2 inch Dynamax silica gel column, 25 mL/min) eluting with 100% ethyl acetate to give 1.2 g (21%) of 4-hydroxy-3-[(Z)-1-oxo-6-octadecenyl]-2(5H)-thiophenone as an oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 5.34 (m, 2H, CH.sub.2 CH.dbd.CHCH.sub.2), 3.98 (s, 2H, CH.sub.2 SC.dbd.O), 2.96 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 2.05 (m, 4H, CH.sub.2 CH.dbd.CHCH.sub.2), 1.68 (m, 2H, CH.sub.2), 1.43 (m, 2H, CH.sub.2), 1.26 (m, 18H, CH.sub.2), 0.88 (t, 3H, J=7.5 Hz, CH.sub.3); IR (Film) 2980, 2900, 2820, 1695 (C.dbd.O), 1675, 1615 cm.sup.-1 ; MS (CI+) 381 (MH+).
EXAMPLE 52
4-Hydroxy-3-[(Z)-8-(2-octylcyclopropyl)-1-oxooctyl]-2(5H)-thiophenone
To 2.2 g of zinc-copper couple in 30 mL of dry ether is added 5.7 mL (16.9 mmol) of methyl oleate and 5.4 mL (70.7 mmol) of diiodomethane. The reaction mixture is refluxed overnight, cooled to room temperature, poured into 1.0N HCl, and extracted three times with ether. The organic layers are combined, washed with brine, dried over MgSO.sub.4, filtered, and concentrated in vacuo to give an oil. The oil is subjected to the conditions described in Example 51 above to give an oil free of starting material. The oil is taken up in a combination of tetrahydrofuran:methanol:water (2:1:1), followed by the addition of 4.0 g of 85% KOH. After 4 hours stirring at room temperature, the solvents are evaporated, the residue taken up in 0.1N KOH, and extracted three times with ether. The aqueous phase is acidified using concentrated HCl and extracted three times with ether. The ether layers are combined, washed with brine, dried over, MgSO.sub.4, filtered, and concentrated in vacuo to give cis-8-(2 -octylcyclopropyl) octanoic acid. Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 2.35 (t, 2H, J=7.0 Hz, CH.sub.2 C=O), 0.6-1.7 (m, 33H).
To a stirring solution of 330 mg (2.84 mmol) of thiotetronic acid in 20 mL of dimethylformamide is added 410 .mu.L (2.94 mmol) of triethylamine and 100 mg (820 .mu.mol) of 4-dimethylaminopyridine at 0.degree. C. Next, 640 mg (3.33 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.0 g (3.38 mmol) of cis-8-(2-octylcyclopropyl)octanoic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over MgSO.sub.4, and concentrated in vacuo. The residue is recrystallized from 5% ethyl acetate/hexanes, filtering to give 250 mg (22%) of 4-hydroxy-3-[(Z)-8-(2-octylcyclopropyl)-1-oxooctyl]-2(5H)-thiophenone, mp 30.degree.-32.degree. C.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.0 (s, 2H, CH.sub.2 SC.dbd.O), 2.96 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 1.65 (m, 2H, CH.sub.2), 1.2-1.4 (m, 12H, aliphatic), 0.88 (t, 2H, J=7 Hz, cyclopropyl), 0.60 (m, 2H, cyclopropyl); IR (KBr) 2910, 2840, 1685 (C.dbd.O), 1620, 1570 cm.sup.-1 ; MS (EI) 394 (M+).
Analysis Calc'd. for C.sub.22 H.sub.38 SO.sub.3 : C, 70.01; H, 9.71. Found: C, 70.13; H, 9.52.
EXAMPLE 53
4-Hydroxy-3-[(Z)-1-oxo-2-(2-pentylcyclopropy)ethyl]-2(5H)-furanone
Cis-nonen-1-ol (10.0 g) is subjected to the cyclopropanation conditions as described in the preparation of Example 52. The resulting residue is taken up in 100 mL of dichloromethane at 0.degree. C., followed by the addition of 800 mg of sodium bromide in 2 mL of water and 150 mg of 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical. To this mixture is added 1.50 g of Aliquot 336 followed by the dropwise addition of 9.2 g of sodium bicarbonate in 230 mL of 5% NaOCl. The aqueous phase is made basic, separated from the organic phase and acidified with concentrated HCl, and then extracted three times with dichloromethane. The combined organic layers are washed with water, dried over MgSO.sub.4, and concentrated in vacuo to give cis-2-(2'-pentylcyclopropyl)acetic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 1.5 (m, 2H, CH.sub.2 C.dbd.O), 0.8-1.6 (m, 15H, aliphatic).
To a stirring solution of 500 mg (5.0 mmol) of tetronic acid in 20 mL of dimethylformamide is added 250 .mu.L (1.79 mmol) of triethylamine and 200 mg (1.80 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 1.13 g (5.89 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.0 g (5.89 mmol) of cis-2-(2'-pentylcyclopropyl)acetic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over MgSO.sub.4, and concentrated in vacuo. The residue is recrystallized from pentane, filtering to give 80 mg (6%) of 4-hydroxy-3-[(Z)-1-oxo-2-(2-pentylcyclopropyl)ethyl]-2(5H)-furanone, mp 55.degree.-58.degree. C.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.67 (s, 2H, CH.sub.2 OC.dbd.O), 2.9 (m, 4H, CH.sub.2 cyclopropyl and O.dbd.CCH.sub.2), 1.1-1.5 (m, 14H, aliphatic), 0.9 (m, 6H, CH.sub.3 and cyclopropyl), 0.70 (m, 2H, cyclopropyl); IR (KBr) 2920, 2840, 1770 (C.dbd.O), 1650, 1610 cm.sup.-1 ; MS (EI) 252 (M+).
Analysis Calc'd. for C.sub.13 H.sub.20 O.sub.4 : C, 66.65; H, 7.99. Found: C, 65.36; H, 7.27.
EXAMPLE 54
4-Hydroxy-3-[(Z)-8-(2-butylcyclopropyl)-1-oxooctyl]-2(5H)-furanone
Methyl myristoleate (1.5 g) is subjected to the cyclopropanation and saponification conditions described above for Example 52 to give cis-8-(2'-butylcyclopropyl)octanoic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 2.35 (t, 2H, J=7.5 Hz, CH.sub.2 C.dbd.O), 0.6-1.8 (m, 25H).
To a stirring solution of 300 mg (3.0 mmol) of tetronic acid in 20 mL of dimethylformamide is added 460 .mu.L (3.21 mmol) of triethylamine and 700 mg (5.74 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 700 mg (3.65 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 820 mg (3.42 mmol) of cis-8-(2'-butylcyclopropyl)octanoic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over MgSO.sub.4, and concentrated in vacuo. The residue was subjected to flash chromatography (acid washed silica gel) eluting with 50% ethyl acetate/hexanes to give 10 mg (10%) of 4-hydroxy-3-[(Z)-8-(2-butylcyclopropyl)-1-oxooctyl]-2(5H)-furanone as a yellow wax.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.68 (s, 2H, CH.sub.2 OC.dbd.O), 2.92 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 0.60-2.0 (m, 15H,cyclopropyl and aliphatic); IR (KBr) 2920, 2850, 1780 (C.dbd.O), 1750, 1650, 1610 cm.sup.-1 ; MS (EI) 322 (M+).
Analysis Calc'd. for C.sub.19 H.sub.30 O.sub.4 : C, 70.77; H, 9.38, Found: C, 70.06; H, 8.86.
EXAMPLE 55
4-Hydroxy-3-[(Z)-1-oxo-5-(2-undecylcyclopropyl)pentyl]-2(5H-furanone
Methyl petroselinate (2.5 g) was subjected to the cyclopropanation and saponification conditions described in Example 52 to give cis-5-(2'-undecylcyclopropyl)pentanoic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 2.35 (t, 2H, J=7 Hz, CH.sub.2 C.dbd.O), 0.6-1.8 (m, 33H, aliphatic).
To a stirring solution of 420 mg (4.20 mmol) of tetronic acid in 20 mL of dimethylformamide is added 640 .mu.L (4.59 mmol) of triethylamine and 160 mg (1.31 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 1.0 g (5.20 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1.5 g (5.06 mmol) of cis-5-(2'-undecylcyclopropyl)pentanoic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over MgSO.sub.4, and concentrated in vacuo. The residue is subjected to flash chromatography (acid washed silica gel) eluting with 50% ethyl acetate/hexanes to give a 400 mg (25%) of 4-hydroxy-3-[(Z)-1-oxo-5-(2-undecylcyclopropyl)pentyl]-2(5H)-furanone as a yellow solid, mp 40.degree.-42.degree. C.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.68 (s, 2H, CH.sub.2 OC.dbd.O), 2.95 (t, 2H, J=7.5 Hz, O.dbd.CCH.sub.2), 0.60-2.1 (m, 20H,cyclopropyl and aliphatic); IR (KBr) 2920, 2840, 1770 (C.dbd.O), 1690, 1600 cm.sup.-1 ; MS (EI) 378 (M+).
Analysis Calc'd. for C.sub.23 H.sub.38 O.sub.4 : C, 72.98; H, 10.12. Found: C, 72.64; H, 8.88.
EXAMPLE 56
4-Hydroxy-3-[(Z)-1-oxo-2-(2-pentylcyclopropyl)ethyl]-2(5H)-thiophenone
Cis-nonen-1-ol (10.0 g) is subjected to the cyclopropanation conditions described in the preparation of Example 52. The resulting residue is taken up in 100 mL of dichloromethane at 0.degree. C., followed by the addition of 800 mg of sodium bromide in 2 mL of water and 150 mg of 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical. To this mixture is added 1.50 g of Aliquot 336 followed by the dropwise addition of 9.2 g of sodium bicarbonate in 230 mL of 5% NaOCl. The aqueous phase is made basic, separated from the organic phase and acidified with concentrated HCl, and then extracted three times with dichloromethane. The combined organic layers are washed with water, dried over MgSO.sub.4, and concentrated in vacuo to give cis-2-(2'-pentylcyclopropyl)acetic acid.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 1.5 (m, 2H, CH.sub.2 C.dbd.O), 0.8-1.6 (m, 15H, aliphatic).
To a stirring solution of 2.0 g (17.2 mmol) of thiotetronic acid in 20 mL of dimethylformamide is added 2.6 mL (18.64 mmol) of triethylamine and 640 mg (5.25 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 4.0 g (20.8 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 3.5 g (20.6 mmol) of cis-2-(2'-pentylcyclopropyl)acetic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over MgSO.sub.4, and concentrated in vacuo. The residue is subjected to flash chromatography (acid washed silica gel) eluting with 10% ethyl acetate/hexanes to give 400 mg (9%) of 4-hydroxy-3-[ (Z)-1-oxo-2-(2-pentylcyclopropyl)ethyl]-2(5H)-thiophenone as an orange oil.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.0 (s, 2H, CH.sub.2 SC.dbd.O), 3.0 (m, 4H, CH.sub.2 cyclopropyl and O.dbd.CCH.sub.2), 1.65 (m, 2H, CH.sub.2), 0.7-1.5 (m, 10H, aliphatic); IR (KBr) 2920, 1685 (C.dbd.O), 1620, 1570 cm.sup.-1 ; MS (EI) 268 (M+).
Analysis Calc'd. for C.sub.13 H.sub.20 SO.sub.3 : C, 62.66; H, 7.51. Found: C, 62.84; H, 7.36.
EXAMPLE 57
4-Hydroxy-3-[1-oxohexadecyl]-2(5H)-furanone
To a stirring solution of 650 mg (6.5 mmol) of tetronic acid in 20 mL of dimethylformamide is added 1.0 mL (7.17 mmol) of triethylamine and 230 mg (1.88 mmol) of 4-dimethylaminopyridine at 0.degree. C. Next, 1.50 g (7.81 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2.0 g (7.81 mmol) of palmitic acid are added and the reaction mixture is stirred overnight at room temperature. The reaction is acidified with 1.0N HCl and extracted three times with ethyl acetate. The combined organic layers are washed with brine, dried over MgSO.sub.4, and concentrated in vacuo. The residue is recrystallized from pentane to give 2.0 g (91%) of 4-hydroxy-3-[1-oxohexadecyl]-2(5H)-furanone, mp 81.degree.-83.degree. C.
Spectral data follows: .sup.1 H NMR (CDCl.sub.3, 400 MHz) .delta. 4.7 (s, 2H, CH.sub.2 OC.dbd.O), 2.9 (t, 2H, J=7.0 Hz, O.dbd.CCH.sub.2), 1.8-0.8 (m, 29H, aliphatic); IR (KBr) 2920, 2840, 1775 (C.dbd.O), 1750, 1660, 1620 cm.sup.-1 ; MS (EI) 338 (M+).
Analysis Calc'd. for C.sub.20 H.sub.34 O.sub.4 : C, 70.97; H, 10.12. Found: C, 70.91; H, 9.73.
EXAMPLE 58
The compounds 5- and 12-hydroxyeicosatetraenoic acid (5-HETE and 12-HETE) and LTB.sub.4 are early arachidonic acid oxidation products in the lipoxygenase cascade, which have been shown to mediate several aspects of inflammatory and allergic response. This is especially true with respect to 5,12-diHETE, which is also denoted as LTB.sub.4 [see Ford-Hitchinson, J. Roy. Soc. Med., 74, 831 (1981)]. Compounds which inhibit the PLA.sub.2 -mediated release of arachidonic acid thereby effectively prevent the oxidation of arachidonic acid to the various leukotriene products via the lipoxygenase cascade. Accordingly, the specificity of action of PLA.sub.2 inhibitors can be determined by the activity of test compounds in this assay, which measures the ability of compounds to inhibit the synthesis of LTB.sub.4 and 5-HETE by rat glycogen-elicited polymorphonuclear leukocytes (PMN) in the presence of exogenous substrate.
The assay is carried out as follows:
Rat polymorphonuclear leukocytes (PMNs) are obtained from female Wistar rats (150-200 g) which receive an injection of 6% glycogen (10 ml i.p.). Rats are sacrificed 18-24 hours post injection by CO.sub.2 asphyxiation and the elicited cells are harvested by peritoneal lavage using physiological saline (0.9% NaCl). The exudate is centrifuged at 400 xg for 10 minutes. The supernatant fluid is discarded and the cell pellet is resuspended to a concentration of 2.0.times.10.sup.7 cells/mL in HBSS containing Ca.sup.++ and Mg.sup.++ and 10 .mu.M L-cysteine.
To 1 mL aliquots of cell suspension, test drugs or vehicle are added, then preincubated at 37.degree. C. for 10 minutes. A23187 (1 .mu.M), [.sup.3 H]-AA (3.0 .mu.Ci/mL) and unlabeled AA (1 .mu.M) are then added and the samples are further incubated for 10 minutes. The reaction is terminated by centrifugation and pelleting cells. Supernatants are then analyzed by HPLC analysis on a 15 cm.times.4.6 mm ID supelcosil LC-18 (Supelco)(3M) column, using a two solvent system at a flow rate of 1.4 mL total flow as follows:
Solvent A: 70:30 17.4 mM H.sub.3 PO.sub.4 :CH.sub.3 CN.
Solvent B. CH.sub.3 CN.
______________________________________Gradient: (system is equilibrated with Solvent A)Time Percent A Percent B______________________________________0 100 015.0 100 020.0 65 3540.0 65 3542.0 10 9050.0 10 9050.1 100 0______________________________________
Percent solvent changes are accomplished in a linear fashion.
Injections: 140 .mu.L of each supernatant is injected directly onto column and .sup.3 H arachidonic acid metabolites are monitored using an on-line radioactivity detector (Ramona, IN/US, Fairfield, N.J.).
Standards: 10.sup.4 -2.0.times.10.sup.4 dpm of eicosanoids of interest are injected in 90 .mu.L EtOH cocktail.
Co-chromatography with standard [.sup.3 H] leukotriene B.sub.4 (LTB.sub.4) in medium of stimulated PMN exposed to drug is compared to that found in medium of stimulated cells exposed to no drug, generating percent inhibition.
Results are expressed as percent inhibition at a given compound dose or as an IC.sub.50 value.
Testing compounds of the invention in this assay gave the following results:
TABLE I______________________________________Compound of % InhibitionExample No. LTB.sub.4 5-HETE______________________________________etodolac 21 (at 0.5 .mu.M)indomethacin 31 (at 10 .mu.M) 1 60 (at 50 .mu.M) 7 +13* (at 10 .mu.M) 8 +10* (at 10 .mu.M) 9 1 (at 10 .mu.M)12 +5* (at 10 .mu.M)23 83 (at 10 .mu.M) 93 (at 10 .mu.M)25 8 (at 10 .mu.M) +81* (at 10 .mu.M)26 82 (at 10 .mu.M) 85 (at 10 .mu.M)______________________________________ *Denotes potentiation of lipoxygenase (LTB.sub.4 or 5HETE synthesis).
EXAMPLE 59
The procedure of Example 58 is also employed for the determination of the extent to which compounds of the invention inhibit the synthesis of the arachidonic acid cyclooxygenase oxidation products PGE.sub.2 and TxB.sub.2.
In this assay, the procedure of Example 58 is carried out as described. However, in order to determine cyclooxygenase activity, the samples are co-chromatographed with authentic reference [.sup.3 H]-TxB.sub.2 or [.sup.3 H]-PGE.sub.2.
The results are calculated as in Example 58 and presented below:
TABLE II______________________________________Compound of % InhibitionExample No. TxB.sub.2 PGE.sub.2______________________________________etodolac 100 (at 0.5 .mu.M)indomethacin 100 (at 10 .mu.M) 1 71 (at 50 .mu.M) 7 +18* (at 10 .mu.M) 8 1 (at 10 .mu.M) 9 31 (at 10 .mu.M)12 9 (at 10 .mu.M)23 8 (at 10 .mu.M) +1* (at 10 .mu.M)25 +8* (at 10 .mu.M) +68* (at 10 .mu.M)26 1 (at 10 .mu.M) +6* (at 10 .mu.M)______________________________________ *Denotes a potentiation of cyclooxygenase (PGE.sub.2 or TxB.sub.2 synthesis).
EXAMPLE 60
The compounds of the invention are tested in an in vitro phospholipase A.sub.2 assay to determine the ability of the compounds to inhibit the biosynthesis of platelet-activating factor and LTB.sub.4 in purified human neutrophils.
This assay is carried out as follows:
Isolation of Human Polymorphonuclear Neutrophils
A leukocyte enriched blood sample obtained from a healthy male donor is procured by leukophoresis using a Haemonetics model 30+ blood processor (Biological Specialties, Inc., Lansdale, Penn.). The top "platelet-rich" layer is removed after a low speed spin (35 xg, 15 min, 25.degree. C.) of the sample. The remaining cell suspension is centrifuged (400 xg, 10 min, 25.degree. C.) to sediment the remaining cells. The supernatant is discarded and the cell pellet resuspended in 120 ml HBSS (without Ca.sup.++ /Mg.sup.++). The cell suspension is subjected to ficoll-hypaque sedimentation (Histopaque 1077, 400 xg, 30 min, 25.degree. C.). Contaminating erythrocytes are lysed by hypotonic shock (1 min). The cells are then washed once with 40 ml of HBSS and resuspended with HBSS (without Ca.sup.++ /Mg.sup.++) to a concentration of 2.5.times.10.sup.7 cells/ml for further use. Greater than 95% purity is obtained, as assessed by microscopic examination.
Platelet-Activating Factor Biosynthesis in Human Polymorphonuclear Neutrophils (PMN)
One ml of human PMN (2.5.times.10.sup.7 cells/ml) is incubated with vehicle or drugs (10 .mu.l) for 10 minutes at 30.degree. C. After preincubation, an equal volume of HBSS (1 ml) containing 2.4 mM CaCl.sub.2, 6 .mu.M calcium ionophore A23187 and 50 .mu.Ci [.sup.3 H]-acetate is then incubated at 30.degree. C. for 15 minutes. An aliquot (100 .mu.l) of the reaction mixture is taken out and mixed with 900 .mu.l of 15% ethanol. LTB.sub.4 is extracted by using solid phase extraction on reverse phase C.sub.18 columns to remove excess [.sup.3 H]-acetate and PAD. The C.sub.18 column is prewashed once with 2 ml of ethanol and water. The sample aliquot is acidified with 0.1N HCl to pH 3 before applying to the column. The column is then washed with 2 ml of water followed by 2 ml of 15% ethanol and 2 ml of petroleum ether to remove excess labeled acetate. The sample is eluted with 2 ml of ethyl acetate. The collected samples are dried with nitrogen and resuspended in 0.5 ml RIA buffer. The quantity of LTB.sub.4 in the sample is obtained from RIA determination. For PAF determination, the reaction is terminated by addition of 5 ml chloroform:methanol:acetic acid (2:1:0.04, v/v/v). [.sup.3 H]-PAF is obtained by Bligh and Dyer extraction. The chloroform phase is removed and dried under nitrogen. The residue is redissolved in 75 .mu.l of chloroform:methanol (80:20, v/v). [.sup.3 H]-PAF is resolved from other phospholipids by TLC on RPC.sub.18 plates with a solvent system of chloroform:methanol:water (2:3:1, v/v/v) and is quantitated using a Berthold automated TLC linear analyzer.
Data presented are the mean +/- s.d. of the values relative to control A23187 stimulated cells for each experiment assayed in triplicate. Percent inhibition when used is calculated as:
% Inhibition=100-[(x.div.Control).times.100].
Dose response analysis is performed by non-linear regression analysis for curve fitting and IC.sub.50 determination.
In this assay, scalaradial, an irreversible inhibitor of PLA.sub.2, isolated from the marine sponge Cacospongia sp. gives an IC.sub.50 of 1.0 .mu.M.
When tested in this assay, the compounds of the invention gave the following results:
TABLE III______________________________________Compound of Dose, % Inhibition Dose, % InhibitionExample No. .mu.M PAF .mu.M LTB.sub.4______________________________________ 1 10 13 10 37 25 32 25 46 50 68 9 50 56 50 10014 10 16 10 30 25 19 25 -5 50 6415 50 21 50 6818 25 53 25 83 50 4720 50 83 50 9522 10 24 10 93 25 49 25 91 50 6725 10 74 50 47 50 9735 10 81.6 50 5 50 98.546 25 7 50 50 50 3947 50 35 50 4748 25 76 25 8650 10 42 25 10051 50 70______________________________________
EXAMPLE 61
The compounds of the invention are tested in an in vitro isolated phospholipase A.sub.2 assay to determine the ability of the test compounds to inhibit the release of arachidonic acid from an arachidonic acid-containing substrate by the action of phospholipase A.sub.2 enzyme from human and non-human sources.
This assay is carried out as follows:
Into a 15 mL polypropylene tube are added the following:
______________________________________Agent Volume, .mu.L Final Conc.______________________________________.sup.3 H-AA E. coli substrate.sup.1 25 5 nmoles PLCaCl.sub.2 (0.1 M).sup.2 5 5 mMTris-HCl (0.5 M) pH 7.5.sup.3 20 100 mMWater.sup.4 25Drug/vehicle.sup.5 1 50 .mu.MPLA.sub.2 25 Volume yielding 12% 100 hydrolysis in 10 min.______________________________________ *pre-incubate at room temperature 30 min prior to substrate addition. .sup.1 Prepared by adding 2 mL deionized and distilled water to 2 mL .sup.3 Harachidonate labeled E. coli (lower count), to which is added to mL of .sup.3 Harachidonate labeled E. coli (higher count) to yield a tota of 5 m substrate (containing 1000 nmoles phospholipid). .sup.2 Stock 0.1 m CaCl.sub.2, required for enzyme activity. .sup.3 Stock 0.5 m TrismaBase. Stock 0.5 M TrismaHCl. Adjust pH to 7.5 (optimum for enzyme). .sup.4 Deionized and distilled water. .sup.5 Stock 10 mM prepared in dimethyl sulfoxide. Make 1:2 dilution with dimethyl sulfoxide and add 1 .mu.L to 100 .mu.L assay tube. .sup.6 Two human PLA.sub.2 enzymes are used: a) Semipurified human platelet acid extract PLA.sub.2 (in 10 mM sodium acetate buffer, pH 4.5). Remove protein precipitate by centrifugation at about 2200 rpm for 10 minutes. b) Purified human synovial fluid.
Incubate the 100 .mu.L reaction mixture for 10 minutes at 37.degree. C. in a shaking water bath. The reaction is terminated by the addition of 2 mL tetrahydrofuran, followed by vortexing. NH.sub.2 columns (100 .mu.g/mL--Analytichem International) are conditioned with 0.5 mL tetrahydrofuran followed by 0.5 mL tetrahydrofuran/water (2 mL:0.1 mL, v/v).
The sample is loaded onto the columns and slowly drawn through them. The hydrolyzed arachidonic acid retained in the columns is eluted therefrom with 1 mL tetrahydrofuran/glacial acetic acid (2%). The arachidonic acid is transferred to scintillation vials and quantitated by .beta.-counting analysis. A "total counts" sample is prepared by pipetting 25 .mu.L .sup.3 H-arachidonate E. coli directly into a scintillation vial to which is added 1 mL tetrahydrofuran. 10 mL aquasol (scintillation cocktail) is added to all samples.
______________________________________Calculations: ##STR14## ##STR15##Activity of Standard Drugs: IC.sub.50 (.mu.M) Human Platelet Human SynovialDrug PLA.sub.2 PLA.sub.2______________________________________Arachidonic Acid 8.6 3.2Monoalide 25.2 0.14______________________________________
When tested in this assay, the compounds of the invention gave the following results:
TABLE IV______________________________________Compound of % Inhibition at 10 .mu.M IC.sub.50 (.mu.M)Example No. HP* HSF** HP HSF______________________________________sulindac 33 34 30.2 1 14.9 35.1 10.8 2 8 (at 50 .mu.M) 27 (at 50 .mu.M) 3 3.5 (at 50 .mu.M) 25 (at 50 .mu.M) 4 10 (at 50 .mu.M) 25 (at 50 .mu.M) 5 0 (at 50 .mu.M) 11 (at 50 .mu.M) 6 0 59 8.7 7 0 98 1.8 8 3 77 10.5 9 +18.5 97.3 1.110 +9.6 3411 6.7 3212 +11 82 2.013 11 3414 44 95 1.7 0.4815 30 77.516 +2.1 5.517 16 918 46 919 83 8820 62 8121 2.5 2422 71 89.8 13.1 2.523 52.7 4424A 45.4 80.724 56.6 79.525 58.3 28.8 26.7 33.426 24.4 +1.727 34.9 +31.228 37.4 12.629 20.6 33.630 37.4 21.031A 47.7 43.231 33.5 34.532 31.6 15.133 37.1 +10.035 68.6 28.4 7.5 1636 25.4 10.337 22.5 +35.738 80.9 18.839 24.4 +20.340A 87.7 74.240 96.5 91.041 12 1342 6.6 2243 29.7 44.444 16.1 37.045 63.0 55.646 16.4 72.047 13.8 34.848 1.1 50.949 1.7 6.750 14.1 55.951 15.2 29.452 8.4 18.653 1.7 0.1______________________________________ *human platelet **human synovial fluid
EXAMPLE 62
The ability of the compounds of the invention to act as inhibitors of the enzymes 5-lipoxygenase and cyclooxygenase is measured in the resident murine peritoneal macrophage assay.
This assay is carried out as follows:
Resident peritoneal macrophages are collected from female Swiss Webster mice (49 days old, 20-25 gms, Buckshire) by lavaging with 7-8 ml Hanks Balanced Salt Solution (HBSS) without Ca.sup.++ and Mg.sup.++ (GIBCO). The lavage fluid from several mice is pooled and centrifuged at 4.degree. C. for 10 minutes at 400 xg. The cell pellet is resuspended in Medium 199 (GIBCO) with HEPES buffer containing 100 .mu.g/ml gentamicin. Two ml of the cell suspension (4.times.10.sup.6 cells) are then plated on 35 mm culture dishes (Nunc).
A macrophage monolayer is established after a 1-1.5 hour incubation of the cells at 37.degree. C. in an atmosphere of 95% O.sub.2 and 5% CO.sub.2. The monolayers are washed 2.times. with 2 ml HBDSS, containing Ca.sup.++ and Mg.sup.++ after which 2 ml Medium 199 supplemented with 10% freshly thawed heat-inactivated fetal bovine serum and 100 .mu.g/ml gentamicin is added for an overnight incubation.
Residual serum and cellular debris are removed from the monolayers by washing 3.times. with 2 ml HBSS containing Ca.sup.++ and Mg.sup.++. Macrophages are preincubated for 5 minutes with 1 ml serum-free M199 containing 10 .mu.l dimethyl sulfoxide (DMSO) vehicle or test compound prior to cell activation with zymosan (100 Mg/ml) or arachidonic acid (AA) (2 .mu.M). After 2 hours, the supernatants are removed and either assayed for LTC.sub.4 and PGE.sub.2 by radioimmunoassay (RIA) directly or stored at -20.degree. C. In all cases, results are expressed as ng metabolite/4.times.10.sup.6 cells.
______________________________________Summary of RIAs used for quantitation of metabolitelevels in zymosan or arachidonic acid stimulatedmouse macrophage culture media. Metabolite Levels Range of (ng/4 .times. 10.sup.6 cells)Metabolite detection (.mu.g/ml) (x .+-. S.E.M., n)______________________________________LTC.sub.4 0.25-16 93.7 .+-. 9.9 (34)PGE.sub.2 0.027-20 30.90 .+-. 1.93 (39)______________________________________
Calculations
Raw data (dpm) may be stored directly onto an "Autostart" tape using the HP85 in room C-096. The raw data are converted to ng metabolite/4-10.sup.6 cells using the standard curve by a "RIANAL" program (HP85) or a "NONLIN" program (HP9816). Results are then expressed as percent inhibition of zymosan induced, leukotriene or prostaglandin synthesis (control) using the equation: ##EQU1##
REFERENCE COMPOUNDS
The compounds used are listed below.
IC.sub.50 values of reference 5-lipoxygenase and/or cyclooxygenase inhibitors.
______________________________________IC.sub.50 .mu.M (95%) Confidence limits)Compound LTC.sub.4 PGE.sub.2______________________________________BW 755c 0.21 1.04 (0.10, 0.42) (0.73, 1.49)ETYA 0.44 1.26 (0.36, 0.53) (0.99, 1.60)Indomethacin >50 0.002 (0.001, 0.003)NDGA 1.87 2.15 (0.22, 15.57) (1.15, 4.04)______________________________________
When tested in this assay, the compounds of the invention exhibited the following levels of enzyme inhibition:
TABLE V__________________________________________________________________________ PGE.sub.2 LTC.sub.4Compound of zymosan AA zymosan AAExample No. Dose .mu.M % Inhibition IC.sub.50 Dose .mu.M % Inhibition Dose .mu.M % Inhibition IC.sub.50 Dose %__________________________________________________________________________ Inhibition1 3.0 2.09 0.2 0.3 4 0.2 0.3 814 4.0 5 38 5 6115 0.35 0.4522 0.2 0.218 0.14 0.3 -17 0.16 0.3 -625 0.2 0.6 -338 0.25 0.6 -419 0.5 68.4 0.5 37.520 0.7 1.0 1 0.7 1.0 -5130 1.0 45.8 1.0 41.535 0.15 0.338 1.0 75.8 1.0 46.440 37 0.8 87 0.841 0.1 29 0.1 61 0.5 97 0.5 9042 0.1 -8 0.1 58 0.5 97 0.5 7043 0.1 54 0.1 71 0.5 92 0.5 9644 0.1 -20 0.1 58 0.5 88 0.5 9445 0.2 45 0.2 34.546 0.1 51.9 0.1 0.5 5 0.3 0.5 -947 0.1 48.4 0.1 0.3 -26 0.1 46.2 0.1 0.3 948 0.1 71.8 0.3 -16 0.1 25.3 0.3 -5 0.03 25.4 1.0 77.149 0.5 -11.8 0.5 -19.750 0.1 21.6 0.5 -35 0.1 38.8 0.5 -33 0.5 70.1 0.5 48.151 0.1 0.5 0.5 -795 0.1 21.1 0.5 -4 0.5 91.3 0.5 59.454 0.1 49.5 0.1 59.355 0.1 39.2 0.1 80.3 0.5 74.3 0.5 64.2__________________________________________________________________________
EXAMPLE 63
The ability of the compounds of the invention to inhibit paw edema induced by the exogenous administration of PLA.sub.2 is measured in the in vivo PLA.sub.2 murine paw edema assay.
The assay is carried out as follows:
Non-fasted, male CD-1 mice (8 weeks old; 31-36 grams) are placed in plastic boxes in groups of six. The right hind paw volume is measured using mercury plethysmography (zero time). Compounds are dosed orally (0.5 mL of 0.5% Tween-80) 1 or 3 hours prior to PLA.sub.2 injection or intravenously (0.2 mL in 0.3% dimethylsulfoxide/saline) 3 minutes prior to PLA.sub.2 injection. A solution of purified PLA.sub.2, from the diamond back cotton mouth snake (A. piscivorus piscivorus) is prepared in saline at a concentration of 6 .mu.g/mL. Fifty (50) .mu.L (0.3 .mu.g) of this PLA.sub.2 solution is injected subcutaneously into the right hind paw with a plastic 1 mL plastic syringe (27 gauge, 1" needle). Paw volume of the injected paw is measured again at 10 minutes, 30 minutes and 60 minutes after PLA.sub.2 injection. Animals are euthanized with CO.sub.2 at the completion of the study.
The paw edema is calculated by subtracting the zero time volume from the volume recorded at each time period. Mean paw edema for each treatment group is then calculated and expressed as (.mu.L.+-.S.E.). Drug effects are expressed as a percent change from control (vehicle) values. Statistical significance is determined by a one-way analysis of variance with LSD comparison to control (p<0.05). ED.sub.50 's are determined using repression analysis.
The activity of standard drugs in this assay is as follows:
______________________________________ ED.sub.50 mg/kg p.o.Compound at +10 min.______________________________________Cyproheptadine 3.1BW755C 50Dexamethasone* 10Naproxen 18Aristolochic Acid** Not ActiveLuffarrellolide** Not Active______________________________________ *p.o. 3 hr. **Some activity (30% inhibition) only when coinjected with enzyme.
When tested in this assay, the compounds of the invention gave the following results:
TABLE VI______________________________________Compound of Dose % Change in EdemaExample No. mg/kg 10 min 30 min 60 min______________________________________indomethacin 10 (p.o.)** -32 -31 -42 1 10 (i.v.)* -51 -43 -23 100 (p.o.) -36 -29 -29 7 10 (i.v.) -19 +39 +15 100 (p.o.) -24 -4 -9 9 10 (i.v.) -48 -42 -48 100 (p.o.) -1 +1.5 +7.112 10 (i.v.) -11 -19 +24 100 (p.o.) -27 -33 -223 1 (i.p.)*** -- -41 -20 10 (i.p.) -- -48 -2825 1 (i.p.) -50 -65 -1 10 (i.p.) -38 -44 -18 200 (p.o.) -40 -33 -4326 1 (i.p.) -- -62 -38 10 (i.p.) -- -67 -43______________________________________ *intravenous **peroral ***intraperitoneal
The results show that the compounds of the invention are effective in vivo in inhibiting edema induced by the exogenous administration of snake venom PLA.sub.2.
EXAMPLE 64
The compounds of the invention are evaluated for their ability to inhibit the lipoxygenase and/or cyclooxygenase pathways of arachidonic acid metabolism in the in vivo murine zymosan peritonitis assay.
This assay is carried out as follows:
Male CD-1 mice (8 weeks old) are placed in plastic boxes in groups of six. Animals are injected with 1 mL i.p. of either 1% zymosan in pyrogen free 0.9% saline or saline (unstimulated control). Compounds are dosed orally 1 hour prior to zymosan injection. Twenty minutes after zymosan injection, the mice are asphyxiated by CO.sub.2 inhalation and the peritoneal cavity is lavaged with 2 mL ice cold Hanks Balanced Salt Solution (HBSS) without CaCl.sub.2, MgSO.sub.4. 7H.sub.2 O and MgCl.sub.2.6H.sub.2 O. Peritoneal lavage fluid from each mouse is removed by syringe and placed in 5 mL plastic test tubes put on ice and volume is noted. Preparation of samples for evaluation by ELISA is as follows: Samples are centrifuged at 800 xg for 15 minutes; 1 mL of the supernatant is added to 8 mL ice cold methanol and kept at -70.degree. C. overnight to precipitate protein; and samples are then centrifuged at 800 xg for 15 minutes, followed by a drying procedure in a Savant speed vac concentrator. The samples are reconstituted with 1 mL ice cold ELISA buffer and stored at -70.degree. C. until assayed. The assay for eicosanoids (LTC.sub.4 and 6-keto-PGF.sub.1.alpha.) is performed according to conventional ELISA procedures.
Compounds to be tested orally are suspended in 0.5% Tween 80. Compounds to be tested intraperitoneally are suspended in 0.5% methylcellulose in 0.9% saline.
The total metabolite level in lavage fluid/mouse is calculated and the significance is determined by a one-way analysis of variance with LSD comparisons to control (p.ltoreq.0.05). Drug effects are expressed as a percent change from control values.
The activity of standard drugs in this assay is as follows:
______________________________________ ED.sub.50 mg/kg p.o.Compound LTC.sub.4 6-keto-PGF.sub.1.alpha. /TxB.sub.2______________________________________BW755C <10 22.0Phenidone 24.0 <30.0Indomethacin Not Active 0.126Ibuprofen Not Active 7.0______________________________________
When tested in this assay a compound of the invention and the anti-inflammatory compound etodolac gave the following results:
TABLE VII______________________________________Compound of Dose % InhibitionExample No. mg/kg LTC.sub.4 6-keto-PGF______________________________________indomethacin 10 (p.o.)* +251 100 (p.o.) -58 -58**______________________________________ *perorally administered **negative values denote potentiation
EXAMPLE 65
The ability of the compounds of the invention to inhibit inflammatory responses is examined in the in vivo arachidonic acid (AA)/12-o-tetradecanoylphorbol acetate (TPA)-induced murine ear edema assay.
This assay is carried out as follows:
Swiss Webster female mice (Buckshire), approximately 8 weeks old are placed into plastic boxes in groups of six. Eight groups of mice receive AA topically on the right ear, and another 8 groups receive TPA topically on the right ear. AA and TPA are dissolved in acetone at concentrations of 100 mg/ml and 100 .mu.g/ml respectively. The phlogistics are applied to the right ear by the means of an automatic pipet. Volumes of 10 .mu.l are applied to the inner and outer surfaces of the ear. Each mouse receives either 2 mg/ear AA or 2 .mu.g/ear TPA. The left ear (control) receives acetone delivered in the same manner. Topical and subcutaneous dosing regimens are as follows 1) drugs are given 30 minutes prior to AA treatment and 2) drugs are given 30 minutes after treatment with TPA.
Measurements are taken with Oditest calipers, 0-10 mm with 0.01 graduations. The right and left ears are measured after 1 hr AA-induced inflammation and 4 hours after TPA-induced inflammation.
Calculations
The difference between right and left ear thickness is calculated and the significance is determined by a one way analysis of variance with Dunnett's comparisons to control (P=0.05). Drug effects are expressed as a percent change from control values: ##EQU2##
Activity of standard drugs:
______________________________________ Oral ED.sub.50 (mg/kg)Drug (AA) (TPA)______________________________________BW755c 65 88Phenidone 85 235Indomethacin inactive at 10 inactive at 10______________________________________
The results for compounds of the invention tested in this assay are presented in Table VIII.
TABLE VIII______________________________________TPA - Route of Drug AdministrationCom- Topical Subcutaneouspound % %of Exam- Inhibition Inhibitionple No. vehicle at 200 .mu.g IC.sub.50 at 50 m/kg ED.sub.50 mg/kg______________________________________ 1 acetone 31 250 23 9 acetone 42 190 7 EPW 56 2814 acetone 28 150 21 118 EPW 51 3715 acetone 3418 acetone 1 37 EPW 53 2822 acetone 40 450 3625 acetone 30 64 8620 acetone 80 160 76 31 EPW 8135 acetone 3938 acetone 23 acetone 4546 acetone 41 110 73 46 EPW 8247 acetone 74 77 EPW 6948 5850 78 1951 55 76______________________________________
EXAMPLE 66
The compounds of the invention are further tested in the rat carrageenan paw edema assay to determine their ability to inhibit the acute inflammatory response.
This assay is carried out as follows:
140-180 g Male Sprague-Dawley rats, in groups of 6 animals are injected subcutaneously in the right paw with 0.1 mL of 1% carrageenan at zero time. Mercury plethysmographic readings (mL) of the paw are made at zero time and 3 hours later. Test compounds are suspended or dissolved in 0.5% methylcellulose and given perorally 1 hour prior to carrageenan administration.
The increase in paw volume (edema in mL) produced by the carrageenan is measured. Paw edema is calculated (3 hour volume minus zero time volume), and percent inhibition of edema is determined. Unpaired Student's t-test is used to determine statistical significance.
The activity of standard drugs in this assay is as follows:
______________________________________ Oral ED.sub.50Drug (95% C.L.) mg/kg______________________________________Indomethacin 3.7 (0.6, 23.8)Aspirin 145.4 (33.1, 645.6)Phenylbutazone 26.2 (2.3, 291.0)______________________________________
When tested in this assay, a compound of the invention and the anti-inflammatory drug etodolac gave the following results:
TABLE IX______________________________________Compound of Dose* % InhibitionExample No. (mg/kg) 50 mg/kg (peroral)______________________________________1 50 406 50 317 50 479 100 4112 50 30______________________________________ *administered perorally
The results show that the compounds tested have activity in the rat carrageenan paw edema assay, evidencing an effect on the acute inflammatory response.

Number	Date	Country
4969659	Jul 1974	JPX
51-1633	Jan 1976	JPX
62-19582	Jan 1987	JPX
1276061	Jun 1972	GBX

Tetronic and thiotetronic acid derivatives as phospholipase A.sub.2 inhibitors

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Non-Patent Literature Citations (11)

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