MXPA96005760A - Synthesis of 4-alkenil or 4-alcanil-2-hydroxitetronic acids optically pu - Google Patents

Synthesis of 4-alkenil or 4-alcanil-2-hydroxitetronic acids optically pu

Info

Publication number
MXPA96005760A
MXPA96005760A MXPA/A/1996/005760A MX9605760A MXPA96005760A MX PA96005760 A MXPA96005760 A MX PA96005760A MX 9605760 A MX9605760 A MX 9605760A MX PA96005760 A MXPA96005760 A MX PA96005760A
Authority
MX
Mexico
Prior art keywords
further characterized
furanone
dihydroxy
carbon atoms
monocytes
Prior art date
Application number
MXPA/A/1996/005760A
Other languages
Spanish (es)
Other versions
MX9605760A (en
Inventor
Mantri Padmaja
T Witiak Donald
Original Assignee
The Ohio State University Research Foundation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US08/245,615 external-priority patent/US5504107A/en
Application filed by The Ohio State University Research Foundation filed Critical The Ohio State University Research Foundation
Publication of MX9605760A publication Critical patent/MX9605760A/en
Publication of MXPA96005760A publication Critical patent/MXPA96005760A/en

Links

Abstract

The present invention relates to an optically pure compound of the general formulas Ia or Ib: wherein R is an alkenyl group of 2-20 carbon atoms or an alkanol group of 9-20 carbon atoms and when an alkenyl group contains one or more degrees of unsaturation or a physiologically acceptable salt of

Description

SYNTHESIS OF 4-ALOUENIL OR 4-ALCANIL-2-HYDROXITETRON COS ÓPTICALLY PURE ACIDS BACKGROUND OF THE INVENTION The present invention relates generally to methods for the synthesis of optically pure 4-alkenyl- or 4-alkanyl-2-hydroxytetronic acid compounds of aci-reductone, acid. The aci-reductone- compound of 4- (4-** "" chlorophenyl) -2-hydroxytetronic acid (CHTA), possesses antilipidemic and antiaggregation properties that differ from those of classical phenoxyacetic acids as described in Witiak and other J Med. Chem., 1988, 31: 1437-1455 and Kamanna et al., Lipids, 1989, 24: 25-32. Although unsubstituted 2-alkanyl- and 2-acyl-tetronic acids are frequently found in nature, the 2-hydroxy-substituted reduction oxide system is found only in vitamin C and its closely related kinship compounds (iso-ascorbic acid, erythroascorbic acid) and derivatives, and the antibiotic macrolide chlorothricin. The anti-aggregating activities of 2-hydroxytetronic acid-reductone compounds such as CHTA are of interest since blood platelets are involved in the genesis of atherosclerosis. Acy-reductones of 2-hydroxytetronic acid inhibit the aggregation of human platelets induced by cyclooxygenase (CO) and the secretion of [1 C] serotonin in a concentration-dependent manner at equivalent doses, as reported by Witiak et al., J. Med. Chem., 1982, 25: 90-93. The compound CHTA inhibits the function of platelets by inhibiting enzymes and ROS clearance mechanisms that ultimately result in the blockage of thromboxane A2 synthesis. • Reducing oxide analogues such as 2-hydroxytetronic acid, can function as antioxidants in membranes and interfere with free radical procedures involved in the biosynthetic processing of cyclic prostaglandin endoperoxides (PGG2 and PGH2), and subsequently thromboxane A2 from aci-reductones, such as CHTA, possess numerous biological properties and potentially have many therapeutic applications. They present antilipidemic and antithrombotic activities and increase the effectiveness of the activity of killer cells activated by lymphokine promoted by IL-2 (LAK) in human peripheral blood mononuclear cells. (PBMC). The aci-reductones inhibit the aggregation of platelets induced by AA dependent on CO. See Witiak et al., J. Med. Chem., 1982, 25: 90-93; Witiak et al., 1988, J. Med. Chem., 1986 31: 1437-1445 and Witiak et al., J. Med. Chem. 1986, 29: 2170-2174. A positive linear free energy ratio is observed between the parameters of enzyme inhibition and calculated hydrophobicity (D). In this way, 4-biphenyl and 4- (4'-chlorodiphenyl) -2-hydroxytetronic acids possess an estimated 'D' of 1.96 and 2.67 and inhibit platelet aggregation and ~ induced by AA with CI5QS of 135 and 44 μM , respectively. The 4-aryl-2-hydroxytetronic acids potentiate LAK activity induced by IL-2. This activity is related in part to the inhibition of CO. The highly tumoricidal lymphocytes induced IL-2 have therapeutic potential in the treatment of cancers for which conventional antineoplastic therapy is not useful. Activation by-products of IL-2, PGE2, and reactive oxygen species (ROS) such as superoxide anion radical, revoke LAK activity.Ac-reductones such as 4-aryl-, 4-alkanyl- and 4,4-spiroalkanyl-2-hydroxytetronics inhibit CO and PGE2 production and clear ROS, thereby improving LAK activity induced by IL-2, 4-hour normal 51Cr release tests, improvement in LAK activity observed is comparable to the combined synergy obtained using the CO inhibiting indomethacin and the superoxide desmutase of / (SOD) and catalase purifying ROS. See Triozzi and others, Int. J. Immunopharmac. , 1993, 15: 47-54 and Witiak et al., Am. Canc. Res. Mtg., 1993, Florida. In this way, the acyproducts may be useful for enhancing cancer therapy with IL-2. Aci-reductones are also antilipidemic and reduce total cholesterol, triglycerides, VLDL, and LDL in rats fed cholesterol / cholic acid. These compounds have been found to reduce apoB in VLDL in vivo and inhibit copper catalyzed LDL oxidation in vitro. See, Witiak and d. others, J. Med. Chem., 1982, 25: 90-93, (1982); Witiak et al., Current Chem. Ther., 1988, 15: 41-62 (1988) and Witiak et al., J. Med. Chem., 1988, 31: 1437-1445. Free radicals play an important role in toxicities induced by drugs and xenobiotics and activate molecular oxygen to superoxide and other ROS including hydrogen peroxide and hydroxyl radical. Defense mechanisms, including enzymes such as SOD and "catalase" and radical scavengers such as glutathione, retinoic acid and ascorbic acid protect proteins and nucleic acids against free radical toxicities by extinguishing ROS Unsuitable ROS protection results in ischemia. myocardium, photosensitivity, radiation sensitization, emollysis of red blood cells and atherosclerosis It has been found that 4-aryl-2-hydroxytetronic acids have antioxidant efficiencies similar to probucol and Á-tocopherol. ^ others, "Trends in Medicinal Chemistry", pp. 243-256, Blackwell Scientific Publications: Oxford, 1990. Synthesis for 2-hydroxytetronic acids other than ascorbic acid have been reviewed by Haynes and Plimmer in "Tetronic Acids", "Ouart. Rev., 1960, 292-315, and by Shank," Reductones " , Synthesis, 1972, 176-190.The 2-hydroxytetronic acids have generally been prepared using three different routes: (1) hydroxyl group insertion at the 2-position of the corresponding tetronic acid core, (2) Claisen intramolecular ester cyclization of subatituted glyoxylate; (3) cyclization promoted by 2,4-dihydroxy-3-ketobutanoate base, Witiak and Tehim, J. Org. Chem., 1987, 52: 2324-2327 synthesized spiro-2-hydroxytetronic acids of 5 and 6 members using conversion of propargyl alcohol to methyl spiroteotonates by treatment with sodium methoxide The hydroxylation attempted in position 2 by reaction and reaction with dibenzoyl peroxide provided only a 6% yield of corresponding 2-benzoyloxytetronic acid. However, the 2-hydroxyl group was introduced in good yields by lithiation using lithium diisopropylamide (LDA), boronate ester formation [B (eO) 3J and oxidative hydrolysis (AcOH, H2O2). The methyl 2-hydroxytetronate was converted to the corresponding aci-reductone by stirring the 48% HBr at 45 ° C for 12 hours. Ireland and Thompson, J. Org. Chem., 1979, 44: 3041-3052, used the condensation of and Claisen for the construction of 2-hydroxytetronic acids. Witiak and Tehim, J. Org. Chem., 1987, 52: 2324-2327 prepared 5-and 6-member spiro-2-hydroxytetronic acids using strategies developed by Ireland and Thompson, mentioned above, this method was superior for using hydroxyl group insertion methods because fewer steps were needed and the global returns were higher. For example, the intramolecular Claisen cyclization of readily prepared methoxy or benzyloxy thiocarboxylate intermediates using LDA or lithium hexamethyldisilazide (LiHMDA) at -78 ° C occurred in higher yields. The resulting 2-methoxytetronic acids underwent deprotection by acetylation and subsequent reaction with BBr3, while 2-benzyloxytetronic acids were convertible to target 2-hydroxytetronic acids by transfer hydrogenation. Witiak and Tehim, J. Org. Chem., 1990, 55: 1112-1114 developed the first synthesis for optically pure 4-phenyl-2-hydroxytetronic acid using Claisen cyclization and kinetically controlled conditions. The 2-benzyloxymethoxyacetate derivative of the corresponding methyl mandelate underwent said cyclization at -100 ° C using the non-nucleophilic, sterically blocked base, lithium disyclohexylamide (LiDCyA). Deprotection of subsequent benzyl group of tetronic acid generated the desired compound in low overall yields; 12% for both steps. U.S. Patent No. 5,095,126 and U.S. Patent Application No. 07 / 847,295 relate to the preparation of optically pure stereochemically labile 4-substituted-2-hydroxytetronic acid compounds.
BRIEF DESCRIPTION OF THE INVENTION The present invention relates to optically pure alkenyl or 4-alkanyl-2-hydroxytetronic acids and processes for their preparation. The chiral approach of the present invention utilizes a Claisen condensation of 2- [(2-allyloxy) acetyloxy] acetate methyl-2-alkanyl or 2-alkenyl substituted to generate 2-alleloxytetronic 4-alkanyl or 4-alkenyl substituted acids. The deprotection provides aci-reductonas objective. The invention further relates to methods for using said optically pure compounds as potent platelet aggregation inhibitors and pharmaceutical compositions therefor. The invention also relates to the pharmaceutical use of said compositions for the treatment and / or prevention of coronary artery diseases, platelet aggregation and thrombosis, and / or prevention of atherosclerosis.
DETAILED DESCRIPTION OF THE INVENTION In a first embodiment, the present invention relates to optically pure 4-alkenyl or 4-alkanyl-2-hydroxytetronic acid compounds of the general formulas Ia or Ib. (la) A (Ib) wherein R is an alkenyl group of 2-20 carbon atoms or an alkanyl group of 9-20 carbon atoms, and when an alkenyl group contains one or more degrees of unsaturation. In one aspect of the composition, the present invention encompasses novel pharmaceutical compositions comprising the optically pure compound of the general formulas la and Ib, together with a physiologically acceptable carrier or excipient, in an amount sufficient to have antilipidemic or antiplatelet activities in a animal or patient The compounds and their compositions of the present invention are therefore useful in the treatment or prevention of atherosclerotic disorders As used herein, the term "alkenyl group of 2 to 20 carbon atoms containing one or more degrees of unsaturation "means an organic alkanyl group containing one or more double bonds and which may be optionally substituted by one or more halogen groups, '- * - lower alkanyl, alkoxy, aromatic or heteroaromatic.
Examples of unsubstituted alkenyl groups include those such as 3-octadecenyl, 3, 6, 9, 12-octadecatetraenyl and the like. Examples of alkanyl groups of 9 to 20 carbon atoms include hexadecanyl, heptadecanyl, octadecanyl, nonadecanyl and eicosanyl as well as their corresponding branched chain analogues. Examples of substituted lower alkenyl groups include those such as halogen substituted alkylene, e.g., alkenyl substituted with fluorine, chlorine, bronze and iodine; alkenyl substituted with alkanyl, e.g., methanyl-, ethanil and similar alkenyl; and alkenyl substituted with alkoxy, e.g., methoxy, ethoxy and similar-alkenyl. As used herein, the term "lower alkanyl" means straight or branched chain saturated aliphatic hydrocarbon groups, preferably containing from 1 to 6 carbon atoms. Representative of said groups are methyl, ethanyl, isopropanol, isobutanyl, butanyl, pentanyl, hexanyl and the like. The term "alkoxy" means a lower alkanyl group attached to the rest of the molecule by oxygen. Examples of alkoxy are methoxy, ethoxy, propoxy, isopropoxy and the like. The term "aryl" means phenyl or benzyl, optionally substituted by one or more halogen atoms, e.g. , fluorine, chlorine, bromine or iodine, or lower alkanyl groups. A second embodiment of the present invention relates to a process for making optically pure 2-hydroxytetrone-substituted acid compounds of the formula I: (Ic) (Id) wherein R 'is an alkenyl or alkanyl group of 2-20 carbon atoms, and when an alkenyl group contains one or more degrees of unsaturation or an aryl group. This process comprises: (a) coupling of α-allyloxy acetic acid with the optically pure alkanyl ester of formula II or its corresponding isomer, wherein R 'is as defined above and ale is a lower alkanyl group of 1 to 6 carbon atoms in the presence of DCC (N, N-dicyclohexylcarbodiimide) and an acid acceptor such as 4-pyrrolidinopyridine or dimethylaminopyridine, to obtain an α-allyloxy acetyl ester of the formula III or its corresponding isomer, wherein ale and R 'are as defined above; (b) cyclization of the allyl ester of the formula III with LiHMDA to give the allyloxy aci-reductone of the formula IV wherein R 'is as defined above, or its corresponding isomer; (c) isomerization of the allyloxy acyl-reductone of the formula IV with hydrogen and an iridium catalyst to give the corresponding ether-propenyl of the formula V or its corresponding isomer, wherein R 'is as defined above; and (d) hydrolysis of the enol ether of the formula V, or its corresponding isomer with aqueous acid, to give the desired compound of formula I. The coupling of step (a) is typically conducted in an anhydrous solvent such as methylene chloride, under an inert atmosphere. Preferably, the reaction is conducted at temperatures of about 5 ° C at room temperature, for periods of 6 to 24 hours. The buffering reagents used are preferably DCC or hydroxybenzotriazine (HOBT) or the like in combination with a catalytic nucleophile such as 4- (N, N-dimethylamino) pyridine (DMAP), 4-pyrrolidinopyridine or others typically used for this purpose. The cyclization of step (b) is typically conducted in an anhydrous solvent such as tetrahydrofuran under an inert atmosphere. Preferably, this reaction is conducted at temperatures of about -78 ° C for periods of 1 to 4 hours. LiHMDA is typically used as a sterically blocked non-nucleophilic base, but other similar sterically blocked bases can be used. The isomerization of step (c) is conducted preferably in an anhydrous solvent, such as tetrahydrofuran, under an inert atmosphere. Typically, the reaction is conducted at room temperature, with reaction times of about 1 to 4 hours. The hydrolysis of step (d) is conducted with an aqueous acid, preferably 50% acetic acid, at reflux temperature. The invention also provides the pharmaceutical compositions comprising the optically pure compounds of the general formula I, as well as their physiologically acceptable salts (such as, for example, Na +, K +, NH 4 +. The compounds of the invention have antilipidemic and angling activity and Accordingly, the invention further provides optically pure compounds of the general formula I and their physiologically acceptable salts for use in the therapy or prophylaxis of atherosclerotic disorders.When they are tested in accordance with the methods of the invention, they are useful in the treatment or prevention of atherosclerotic disorders. described in the art, the isomers (S) of the formula Ib having the formula where R is as defined above, it has been found that they possess markedly superior properties when compared to their corresponding (R) isomers. The R and S enantiomers are tested as inhibitors of aggregation induced by arachidonic acid in a human plasma rich in platelets. Data for individual experiments (2 separate donors) are given as PIC50 (logarithmic molar inhibitory concentration of each drug that blocks aggregation to arachidonic acid by 50%). The inhibitors were preincubated for 1 minute before the addition of arachidonic acid (200-400 μm). Changes in light transmission were measured as an aggregation index and quantified after 4 minutes. These properties are summarized in the following Tables 1 and 2: TABLE 1 (R) -3,4-dihydroxy-5- [(all-Z) -3,6,9, 12-octadecatetraenyl] -2 (5H) -furanone (μM) Empirical data Data -lxLOG (0.000001x 764. 500 3.117 538.000 3.269 657.000 3.182 653.167 AVE 3.189 113.299 STD 0.077 65.413 SEM 0.044 17.346 CV 2.400 3 N 3 371.695 -95% CL 2.999 934.639 + 95% CL 3.380 Retrotransformed / data: (geometric mean) 646.510 AVE 417.291 -95% CL 1001.640 + 95% CL TABLE 2 (S) -3,4-dihydroxy-5 [(all-Z) -3,6,9,1-octadecatetraenyl] -2 (5H) -furanone (μM) Empirical data Data -lxLOG (0. OOOOOlx 0-319 6,496 4,300 5,367 2,200 5,658 2,273 2,223 AVE 5,840 1,992 STD 0.587 1,150 SEM 0.339 87,616 CV 10,043 3 N 3 -2,675 -95% CL 4,383 7,221 + 95% CL 7,297 Retrotransformed / data: (geometric mean) 1,445 AVE 0.050 -95% CL 41.405 + 95% CL Additionally, testing on standardized sieves that examine a broad spectrum of human monocyte activities indicates that the compounds of this invention possess a spectrum of activity that makes them useful in the treatment of pathologies involving acute and chronic inflammation. These tests are as described below and Table 3 below indicates the test results in these tests for the compound respectively.
TEST: TUMOR NECROSIS FACTOR-Á (FNT-Á) This test determines the effect of the test compounds on the production of purified human monocytes TNF-A. The compounds are tested for their ability to down-regulate the production of TNF-A in activated monocytes, or up-regulate the secretion of TNF-A in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log concentrations. • Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of TNF-A in the resulting supernatants is "" quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-well format. The FNT-Á present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-TNF-A antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The levels of TNF-A are determined by interpolation of the resulting heat change in the substrate of a normal curve. All the - "• Supernatants are tested in duplicate with controls and normal values.
TEST: INTERLEUCINE (IL-1β) This test determines the effect of the test compounds on the secretion of IL-1β from purified human monocytes. The compounds are tested for their ability to down-regulate the production of IL-lβ in activated monocytes, or up-regulate the secretion of IL-lβ in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log-rank concentrations. Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of IL-lβ in the resulting supernatants is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-well format. The IL-lβ present in the samples is captured by a specific monoclonal ~~ antibody immobilized on the cavity, and detected with a polyclonal anti-IL-lβ antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The levels of IL-lβ are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
. * "TEST: INTERLEUCINE (IL-6) This test determines the effect of test compounds on the secretion of purified human monocyte IL-6 compounds are tested for their ability to down-regulate the production of IL-6 in activated monocytes, or up-regulate the secretion of IL-6 in unstimulated monocytes The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log./g concentrations Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes The level of IL-6 in the resulting supernatants is quantified in a solid-phase enzyme-linked immunosorbent assay (EIA) performed in a 96-well format. in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-IL-6 antibody conjugated to horseradish peroxidase, followed by a Appropriate course. The levels of IL-6 are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: INTERLEUCINE (IL-8) This test determines the effect of test compounds on the secretion of IL-8 from human monocytes ~ purified. The compounds are tested for their ability to down-regulate the production of IL-8 in activated monocytes, or up-regulate the secretion of IL-8 in unstimulated monocytes. Test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three logig concentrations. Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of IL-8 in the resulting supernatants is quantified in a solid phase enzyme-linked Immunoassay (EIA) performed in a 96-well format. The IL-8 present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-IL-8 antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The levels of IL-8 are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: GRANULOCYTE MACROPHAGE COLONIAL STIMULATOR FACTOR (GM-CSF) This test determines the effect of test compounds on the secretion of GM-CSF from purified human monocytes. The compounds are tested for their ability to down-regulate the production of GM-CSF in monocytes Activated, or up-regulate the secretion of GM-CSF in unstimulated monocytes The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log10 concentrations Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes The level of GM-CSF in the resulting supernatants is quantified in a solid-phase enzyme-linked immunosorbent assay (EIA) performed in a 96-well format. Samples are captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-GM-CSF antibody conjugated to horseradish peroxidase, followed by an appropriate substrate.The levels of GM-CSF are determined by heat exchange interpolation resulting in the substrate of a normal curve All supernatants are tested in duplicate with normal controls and values.
"TEST: INTERLEUCINE-1, receptor antagonist (IL-lra) This test determines the effect of test compounds on the secretion of IL-lra from purified human monocytes The compounds are tested for their ability to down-regulate or up-regulate IL-ra production in unstimulated monocytes and activated monocytes The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log-o- concentrations where appropriate., lipopolysaccharide (LPS) is used to stimulate monocytes. The level of IL-ra in the resulting supernatants is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-well format. The IL-ra present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with an anti-IL-ra polyclonal antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. IL-ra levels are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: TISSUE FACTOR (FT) This test determines the effect of the test compounds on the production of membrane bound tissue (FT) factor of purified human monocytes. The compounds are tested for their ability to down-regulate the production of FT in activated monocytes, or up-regulate the secretion of TF in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three logium concentrations. • Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. In the previous incubation period, the medium is aspirated from the monocytes, and the cells are solubilized in Triton-X 100. The level of TF in the resulting soluble fraction is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-well format The FT present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-FT antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. by interpolation of the resulting heat change in the substrate of a normal curve All supernatants are tested in duplicate with controls and values n Ormales TEST: INTERLEUCINE B4 (LTB4) This test measures the ability of test compounds to modulate the amount of LTB4 produced by the oxygenation of arachidonic acid by 5-lipoxygenase produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of LTB4 from activated monocytes, or up-regulate the secretion of LTB4 from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three logig concentrations. Where it is appropriate, the '* .zimosan is used to stimulate monocytes. The level of LTB4 in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of LTB4 modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PLATELET ACTIVATION FACTOR (FAP) This test measures the ability of test compounds to modulate the amount of PAF produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of FAP from activated monocytes, or up-regulate the secretion of FAP from unstimulated monocytes. Test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three log concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of PAF in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of FAP modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values. , - TEST: PROSTAGLANDINE E2 (PEG2) This test measures the ability of test compounds to modulate the amount of PEG2 produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of PGE2 from activated monocytes, or up-regulate the secretion of PGE2 from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three log ^ Q concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of PGE2 in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of GE2 modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values. IC TEST: TROMBOXANE A2 (TxA2) This test measures the ability of test compounds to modulate the amount of TXA2 (the major cyclooxygenase product) produced by purified human monocytes.
The compounds are tested for their ability to down-regulate the secretion of TXA2 from activated monocytes, or to up-regulate the secretion of TXA2 from non-monocyte monocytes. '""' stimulated. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three logio- concentrations Where appropriate, zymosan is used to stimulate monocytes. The level of TxA2 in the resulting supernatants is quantified in an immunoassay performed in a 96-fold format. cavities The levels of TXA2 modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PEPTIDOLEUCOTRIANS This test measures the ability of test compounds to modulate the amount of peptidoleukotrienes produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of peptidoleukotrienes of activated monocytes, or up-regulate the secretion of peptidoleukotrienes from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three log ^ concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of peptidoleukotrienes in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of peptidoleukotrienes modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PHOSPHOLIPASE A2 (PLA2) This test measures the ability of test compounds to modulate the activity of cellular phospholipase A2 in human monocytes. The compounds are tested for their ability to stimulate phospholipase A2 activity or activate basal activity in unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three log ^ Q concentrations. Where appropriate, zymosan is used to stimulate monocytes. Monocytes with the arachidonyl portion of radiolabeled phospholipids are used in a 96-well format. The release rate of radiolabelled fatty acid in the extracellular medium is quantified by liquid scinitilation counting.All supernatants are tested in duplicate with normal controls and values.
TEST: CHEMOTHERAPY This test measures the ability of test compounds to modulate the chemotactic response of purified human monocytes in response to a chemoattractant. The compounds are - - test four dilutions on a scale of three log ^ Q concentrations. The test compounds are combined with monocytes labeled with a fluorescent indicator and placed in the upper chambers of a chemotaxis chamber, above an optimized concentration of a monocyte chemoattractant placed in the cavities below, and separated by a micropore filter. . After ninety minutes, the migration of cells through the filter is fluorometrically quantified, and expressed in relation to spontaneous and maximum controls. All tests are carried out in triplicate.
TEST: CELL ADHESION This test measures the ability of test compounds to modulate the adhesion of purified human monocytes to cultures of umbilical vein endothelial cells (HUVEC). The HUVEC are grown to come together in 96-well plates. The purified human monocytes are labeled with fluorescent dye. Triplicate cultures of HUVEC and monocytes are treated with four dilutions of test compound on a scale of three concentrations log ^ o <; in addition to stimulated and unstimulated controls. One hour after the treatment, FNT-Á is added to HUVEC cultures to stimulate the production of adhesion molecules. After twelve hours of incubation, the treated monocytes are added to their corresponding HUVEC cultures and incubated for ten minutes. The stimulation index (I.E.) is determined by comparing the fluorescence of treated cultures to untreated controls using a fluorometric reading in a 96-well format.
TEST: This test measures the ability of test compounds to modulate the ability of purified human monocytes to release superoxide anions in response to a zymosan. The compounds are tested at four dilutions on a scale of three log ^ concentrations. Cultures of monocyte compounds are treated with zymosan to stimulate the production of anions. The amount of superoxide anion produced is determined by the ability of the culture to reduce cytochrome C, as measured calorimetrically. The test is carried out in a format of 96 cavities in triplicate and the results are expressed in relation to the unstimulated baseline and maximum controls. The appropriate reference controls are carried out with each test.
"" TEST: PROLIFERATION OF CELLS STIMULATED BY MITOGENS AND REACTION OF MIXED LYMPHOCYTES The purpose of this procedure is to determine the effect of test compounds on the reaction of mixed lymphocytes and the proliferation induced by mittengens of normal human mononuclear cells. Procedure for cell proliferation: Peripheral blood mononuclear cells (PBMNC) are added to a 96-well cell culture plate. The compounds are tested at four dilutions in quadruplicate on a scale of three log concentrations together with negative and inhibitory controls. The test compounds are incubated with the cells for one hour, then phytohemagglutinin (PHA) is added to the reactive test wells and the cultures are incubated for three days. The cultures were then pulsed with the addition of 3H then determined with a Wallace Betaplate counter.
Procedure for two-way mixed lymphocyte reaction: PBMNC from two different donors are added to a 96-well cell culture plate. Four dilutions of test compounds, on a scale of three logio concentrations together with negative controls and inhibitors are then added to the plate and incubated for six days. The crops are pulsed, harvested and subsequently counted as described above. When tested in the above standard tests, a representative compound of formula I, ie, (S) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone, is found to give the following results that are shown later in table 3.
TABLE 3 Key: NE No effect INHIB Inhibition STIM Stimulation CI40 Inhibitory concentration at 40? EC50 Concentration tolerated at 50% TC Tolerated concentration IA Interferes with the test The ability of the compounds of formula I to inhibit the action of several inflammatory cytokines makes them useful in a wide variety of therapeutic methods. Specifically, its ability to mediate or inhibit the actions of TNF-A makes these compounds useful in the treatment of various invasive diseases, infections and inflammatory conditions. Particularly important is the inhibition of the large amount of TNF produced during severe bacterial infections, which can activate a state of shock and tissue damage (septic shock syndrome). A further important use of the compounds of the formula I is to inhibit the TNF that is known to mediate cachexia produced during chronic disease states. Therefore, these compounds are particularly useful in adjunctive therapy for patients with AIDS and cancer to reduce and / or reduce the consequences of cachexia produced during these pathological states. An additional specific treatment method for which the compounds of the present invention are particularly useful is in the treatment of rheumatoid arthritis wherein increased amounts of the inflammatory cytokines, TNF-A and IL-1 are present. Due to its ability to mediate and / or inhibit the action of these cytokines, the inflammation and severity of the disease state can be reduced or eliminated. The compounds of the present invention can also be used in the treatment of multiple sclerosis (MS), Crohn's disease and ulcerative colitis by inhibition and the activity of the inflammatory cytokines that are the basis of these disease states.
TEST: TUMOR NECROSIS FACTOR-Á (FNT-Á) This test determines the effect of the test compounds on the production of purified human monocytes TNF-A. The compounds are tested for their ability to down-regulate the production of TNF-A in activated monocytes, or up-regulate the secretion of TNF-A in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three logite concentrations. Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of TNF-A in the resulting supernatants is quantified in a solid-phase enzyme linked immunosorbent assay (EIA) performed in a 96-well format. The FNT-Á present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-TNF-A antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The levels of TNF-A are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: INTERLEUCINE (IL-1β) This test determines the effect of the test compounds on the secretion of IL-1β from purified human monocytes. The compounds are tested for their ability to down-regulate the production of IL-1d in activated monocytes, or up-regulate the secretion of IL-1β in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log-n concentrations. Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of IL-lβ in the resulting supernatants is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-well format. The IL-lβ present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-IL-1β antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The levels of IL-lβ are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: INTERLEUCINE (IL-6) This test determines the effect of the test compounds on the secretion of IL-6 from purified human monocytes. The compounds are tested for their ability to down-regulate the production of IL-6 in activated monocytes, or up-regulate the secretion of IL-6 in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three "login concentrations." Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. in the resulting supernatants is quantified in a solid-phase enzyme-linked immunoassay (EIA) performed in a 96-well format.The IL-6 present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a anti-IL-6 polyclonal antibody conjugated to '* horseradish peroxidase, followed by an appropriate substrate. The levels of IL-6 are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: INTERLEUCINE (IL-8) This test determines the effect of the test compounds on the secretion of IL-8 from purified human monocytes. The compounds are tested for their ability to down-regulate the production of IL-8 in activated monocytes, or up-regulate the secretion of IL-8 in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three logio- In concentrations where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of IL-8 in the resulting supernatants is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-cavity format. The IL-8 present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-IL-8 antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The levels of IL-8 are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: GRANULOCYTE MACROPHAGE COLONIAL STIMULATOR FACTOR (GM-CSF) This test determines the effect of test compounds on the secretion of GM-CSF from purified human monocytes. The compounds are tested for their ability to down-regulate the production of GM-CSF in activated monocytes, or up-regulate the secretion of GM-CSF in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log concentrations. • Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of GM-CSF in the resulting supernatants is quantified in a solid-phase enzyme-linked immunoassay (EIA) performed in a 96-well format. The GM-CSF present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-GM-CSF antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. The GM-CSF levels are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: INTERLEUCINE-1, receptor antagonist (IL-lra) This test determines the effect of the test compounds on the secretion of IL-lra from purified human monocytes. The compounds are tested for their ability to down-regulate or up-regulate IL-ra production in unstimulated monocytes and activated monocytes. Test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three log-n concentrations. Where appropriate, lipopolysaccharide (LPS) is used to stimulate monocytes. The level of IL-ra in the resulting supernatants is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-well format. The IL-ra present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-IL-ra antibody conjugated to horseradish peroxidase, followed by an appropriate substrate. IL-ra levels are determined by interpolation of the resulting heat change in the substrate of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: TISSUE FACTOR (FT) This test determines the effect of the test compounds on the production of membrane bound tissue (FT) factor of purified human monocytes. The compounds are tested for their ability to down-regulate the production of FT in activated monocytes, or up-regulate the secretion of TF in unstimulated monocytes. The test compounds are incubated for sixteen hours with purified human monocytes at four different dilutions on a scale of three concentrations. Where appropriate, lipopolysaccharide (LPS) is used to stimulate ^ ... monocytes. After the previous incubation period, the medium is aspirated from the monocytes, and the cells are solubilized in Triton-X 100. The level of FT in the resulting soluble fraction is quantified in a solid-phase enzyme-linked Immunoassay (EIA) performed in a 96-cavity format. The FT present in the samples is captured by a specific monoclonal antibody immobilized on the cavity, and detected with a polyclonal anti-FT antibody conjugated to "horseradish peroxidase, followed by an appropriate substrate." FT levels are determined by interpolation of the change of heat resulting in the substrate of a normal curve All supernatants are tested in duplicate with normal controls and values.
TEST: INTERLEUCINE B (LTB4) This test measures the ability of test compounds "- to modulate the amount of LTB4 produced by the oxygenation of arachidonic acid by 5-lipoxygenase produced by purified human monocytes The compounds are tested for their ability to down-regulate the production of activated monocytes LTB4, or up-regulate the secretion of LTB4 from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three logium concentrations. • Where appropriate, zymosan is used to stimulate monocytes. The level of LTB4 in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of LTB4 modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PLATELET ACTIVATION FACTOR (FAP) This test measures the ability of test compounds to modulate the amount of PAF produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of FAP from activated monocytes, or up-regulate the secretion of FAP from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of PAF in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of FAP modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PROSTAGLANDI TO E2 (PEG2) This test measures the ability of test compounds to modulate the amount of PEG2 produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of PGE2 from activated monocytes, or up-regulate the secretion of PGE2 from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three log ^ n concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of PGE2 in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of PGE2 modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: TROMBOXANE A2 (TxA2) This test measures the ability of test compounds to modulate the amount of T A2 (the major cyclooxygenase product) produced by purified human monocytes. The compounds are tested for their ability to down regulate the secretion of TA2 from activated monocytes, or up-regulate the secretion of TXA2 from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three log ^ Q concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of A2 in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of T A2 modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PEPTIDOLEUCOTRIANS This test measures the ability of test compounds to modulate the amount of peptidoleukotrienes produced by purified human monocytes. The compounds are tested for their ability to down-regulate the production of peptidoleukotrienes of activated monocytes, or up-regulate the secretion of peptidoleukotrienes from unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three logig concentrations. Where appropriate, zymosan is used to stimulate monocytes. The level of peptidoleukotrienes in the resulting supernatants is quantified in an immunoassay performed in a 96-cavity format. The levels of peptidoleukotrienes modulated by unknown compounds are determined by interpolation of a normal curve. All supernatants are tested in duplicate with normal controls and values.
TEST: PHOSPHOLIPASE A2 (PLA2) This test measures the ability of test compounds to modulate the activity of cellular phospholipase A2 in human monocytes. The compounds are tested for their ability to stimulate phospholipase A2 activity or activate basal activity in unstimulated monocytes. The test compounds are incubated for ninety minutes with purified human monocytes at four different dilutions on a scale of three logiQ concentrations. Where it is appropriate, zymosan is used to stimulate monocytes. Monocytes with the arachidonyl portion of radiolabeled phospholipids are used in a 96-well format. The percent of radiolabelled fatty acid release in the extracellular medium is quantified by liquid scinitilation counting. All supernatants are tested in duplicate with normal controls and values.
TEST: CHEMOTHERAPY This test measures the ability of test compounds to modulate the chemotactic response of purified human monocytes in response to a chemoattractant. The compounds are tested at four dilutions on a scale of three log ^ n concentrations. The test compounds are combined with monocytes labeled with a fluorescent indicator and placed in the upper cavities of a quimiotaxis chamber, an optimized concentration of a monocyte chemoattractant placed in the cavities below, and separated by a filter of micropore After ninety minutes, the migration of cells through the filter is fluorometrically quantified, and expressed in relation to spontaneous and maximum controls. All tests are carried out in triplicate.
TEST: CELL ADHESION This test measures the ability of test compounds to modulate the adhesion of purified human monocytes to cultures of umbilical vein endothelial cells (HUVEC). The HUVEC are grown to come together in 96-well plates. The purified human monocytes are labeled with fluorescent dye. The triplicate cultures of HUVEC and monocytes are treated with four dilutions of test compound on a scale of three logium concentrations in addition to stimulated and unstimulated controls. One hour after the treatment, FNT-Á is added to HUVEC cultures to stimulate the production of adhesion molecules. After twelve hours of incubation, the treated monocytes are added to their corresponding HUVEC cultures and incubated for ten minutes. The stimulation index (I.E.) is determined by comparing the fluorescence of treated cultures to untreated controls using a fluorometric reading in a 96-well format.
TEST: This test measures the ability of test compounds to modulate the ability of purified human monocytes to release superoxide anions in response to a zymosan. The compounds are tested at four dilutions on a scale of three logig concentrations. Cultures of monocyte compounds are treated with zymosan to stimulate the production of anions. The i * - '- amount of superoxide anion produced is determined by the ability of the culture to reduce cytochrome C, as measured calorimetrically. The test is carried out in a format of 96 cavities in triplicate and the results are expressed in relation to the unstimulated baseline and maximum controls. The appropriate reference controls are carried out with each test.
TEST: PROLIFERATION OF CELLS STIMULATED BY MITOGENS AND REACTION OF MIXED LYMPHOCYTES The purpose of this procedure is to determine the effect of test compounds on the reaction of mixed lymphocytes and proliferation induced by mitogens of normal human mononuclear cells. Procedure for cell proliferation: Peripheral blood mononuclear cells (PBMNC) are added to a 96-well cell culture plate. The compounds are tested at four dilutions in quadruplicate on a scale of three log-g concentrations along with negative and inhibitory controls. The test compounds are incubated with the cells for one hour, then phytohemagglutinin (PHA) is added to the reactive test wells and the cultures are incubated for three days. The cultures were then pulsed with the addition of 3H then determined with a Wallace Betaplate counter. Procedure for two-way mixed lymphocyte reaction: PBMNC from two different donors are added to a 96-well cell culture plate. Four dilutions of test compounds, on a scale of three concentrations l ° 910 together with negative and inhibitory controls are then added to the plate and incubated for six days. The crops are pulsed, harvested and subsequently counted as described above. The compounds of the invention may be formulated in a conventional manner, optionally together with one or more additional active ingredients, for administration by any convenient route, for example, oral, intravenous or intramuscular administration. Therefore, in accordance with another aspect, the invention provides a pharmaceutical composition comprising a compound of the formulas Ia or Ib and / or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier or excipient.
For oral administration, the pharmaceutical composition can take the form of, for example, tablets, capsules, powders, solutions, syrups or suspensions prepared by conventional means with physiologically acceptable excipients. The compounds can be formulated for intravenous or intramuscular administration in dry form for reconstitution before use, or as a sterile solution or suspension. A proposed daily dose on similar pharmacokinetic parameters for CHTA for administration to a man is 10 to 25 mg / kg, for example 1 mg daily to 70 kg, which can be conveniently administered in 1 to 3 doses per day. Of course, the precise dose will depend on the age and condition of the patient. The following examples illustrate the present invention. The melting points were determined in open capillaries with a Thomas-Hoover Uni-Melt apparatus and are not corrected. The infrared spectra were recorded by an RFX-FTIR laser precision analytical spectrometer (model TSI-400). Nuclear magnetic resonance spectra were obtained with either a FT NMR spectrometer model NR / 250, 270 or 500. Tetra-ethylsilane (TMS) in CDC13, acetone-dg or CD3OD was used on the scale of the á scale with peak multiplicities : s, individual band; d, doublet; dd, doublet of doublets; ddd, doublet of doublets of doublets; t, triplet; q, quadriplete; m, multiple bands. The tetrahydrofuran (THF) was distilled from Na / benzophenone-cetyl and CH2CI2 was dried over P2O5. Optical rotations were taken on a Perkin-Elmer model 241 polarimeter using a 10 cm cell, 1 Ml. The mass spectra were acquired with a mass spectrometer Kratos MS25RFA or VG 70-250S. Elemental analyzes were performed by Galbraith Laboratories Inc., Knoxville, TN.
PREPARATION OF STARTING MATERIALS EXAMPLE A 3,6,9, -Pßntadßcatrin-1-ol. In a 1 liter, three neck, flame-dried, round-bottomed flask equipped with a reflux condenser and a rubber spectrum, 3.86 g were placed. (159 mmoles) of magnesium in 250 ml of anhydrous tetrahydrofuran.
Bromoethanes (17.3 g, 1585 mmoles) in 250 ml of tetrahydrofuran were added dropwise under argon, and the reflux rate was controlled with the aid of an ice bath The mixture was heated to reflux and stirred for 1 hour. 3-butyl-1-ol (5.56 g, 79.3 mmol), dissolved in 150 ml of anhydrous tetrahydrofuran, was slowly added dropwise with stirring (2 hours) After the addition, the reaction was heated to reflux. stirring for 90 minutes, 0.5 g (2.63 mmoles) of cuprous iodide (I) was added.After 75 minutes, 9.0 g (39.65 mmoles) of l-bromo-2,5-undecadine dissolved in 150 ml of anhydrous tetrahydrofuran was added. The mixture was refluxed for 12 hours and 0.25 g (1.32 ml) of additional cuprous iodide (I) was added.The mixture was heated to reflux for 7 hours, cooled and quenched with the addition of 400 ml of water with ice saturated with ammonium chloride. After filtration (Celite), the filtrate extracted with 3 x 400 ml of ether. The ether layers were washed with 2 x 300 ml of saturated ammonium chloride solution, 3 x 200 ml of water, 250 ml of brine, dried with 5.75 g (75%) of an unstable yellow oil (room temperature) which it was used immediately in the next IR reaction (Neto ^ pf1) 3365, 2956, 2933, 2225; *? NMR (CDCl,) 5 3.70 (t, J - 6.2 Hz, 2H), 3.17-3.13 (m, 4H), 2.45 (dt, J - 1.5, 5.9, 12.0 Hz, 2H), 2.15 (dt, J - 2.1 , 6.9, 13.9 Hz, 2H), 1.68 (br, 1H), 1.55-1.43 (m, 2H), 1.43-1.26 (ra, 4H), 0.89 (t, J - 6.9 Hz, 3H); HRMS calculated for • C ^ oO (M *) 216.1514, Encont. 216.1519. (3Z, 6Z, 9Z) -3,6, 9-Pentadecatriene-1-ol. 3.6, 9-Pentadecanotriol (5.75 g, 26.6 mmol), 5% palladium on barium saulfate (0.5 g) and 5 drops of 3% quinoline in methanol were added to a 500 ml hydrogenation flask. Hydrogen was collected at an initial pressure of 5.06 kg / cm2 for 30 minutes. The mixture was filtered (Celite) and the filtrate was evaporated in vacuo to yield 5.7 g of crude triene which was purified on silica gel using ethyl acetate hexanes (1: 5) yielding 5.5 (93%) of a light yellow oil. . IR (Net cm "1) 3336 (br), 3012, 2958, 2927, 1652, 719; XH NMR (CDClj) 6 5.60-5.28 (ra, 6H), 3.67 (t, J m 6.4 Hz, 2H), 2.88-2.79 (m, 2H), 2.43-2.33 (m, 2H), 2.12-1.98 (m , 2H), 1.68-1.53 (m, 2H), 1.53-1.23 (m, 6H), 0.89 (t, J = 6.7 HZ, 3H); HRMS calculated for C1SH260 (M *) 222.1984, Encont.222.1990. (3Z, 6Z, 9Z) -l-Bromo-3 / 6,9-pentancatriene-1-ol. To a 250 ml three-necked round bottom flask were added under nitrogen 9.46 g (36.1 mmol) of triphenylphosphine dissolved in 150 ml of anhydrous acetonitrile. After cooling to 0 ° C (ice-salt bath), bromine (5.77g, 36.1 mmol) was added per drops with stirring. The mixture was warmed to room temperature and stirred for 30 minutes. 3, 6, 9-Pentadecanotriol (5.75 g, 36.1 mmoles), dissolved in anhydrous acetonitrile, was added dropwise for about 4 hours. Upon completion of the reaction, the acetonitrile was removed in vacuo and the residue was dissolved in 75 ml of ether. Hexanes were used to precipitate the side product of triphenylphosphorane which was removed by filtration. The crude residue, obtained after concentration of the filtrate in vacuo, was purified on silica gel using ethyl acetate: hexanes (1: 9) as eluent. »IR (Net cm * '1) 2958, 2927, 1652, 1267, 723, * H NMR (CDCl,) d 5.69-5.28 (m, 6H), 3.38 (t, J - 7.1 Hz, 2H), 2.89-2.75 (m, 4H), 2.75-2.52 ( m, 2H), 2.12-1.93 (m, 2H >, 1.49-1.31 (m, 6H), 0.89 (C, J • 6 .8 H2, 3H); l3C NMR (CDCl,) 6 131. 0, 130. 6, 128. 9, 127 .4 (2C), 126.3, 32.2, 31.5, 30.9, 29.3, 27. 3, 25.8, 25 .7, 22. 6, 14 0 / HSMS calculated for C ^ Br (M *) 284. 1139, Encont.284. 1101 Bromide dß (3Z, 6Z, 9Z) -3,6,9-pentane-triptyl-1-triphenylphosphonium - It was treated (3Z, 6Z, 9Z) -l-Bromo-3, 6, -pentadecatriene (7.0 g, 24. 6 mmoles) with 7.5 g (28.6 mmoles) of triphenylphosphine in 25 ml of acetonitrile. The mixture was heated to 70 ° C under controlled atmosphere. After completion of the reaction (salt formation was monitored by CCD, 72 hr) the mixture was dried during 36 hours under reduced pressure to ensure the removal of minimal amounts of acetonitrile. The resulting yellow residue, 13.45 g, was used in the next reaction without further purification: __, IR (Net cm'1) 3010, 2958, 1652, 1191, 723; ** H NMR (CDCl,) d 7.93- 7.61 (m, 15 H), 5.69-5.12 (m, 6H), 4.02-3.91 (m, 2H), 2.69-2.48 (m, 4H), 2.09-1.92 ( m, 2H), 1.89-1.72 (m, 2H), 1.46-1.25 (m, 6H), 0.88 (t, J »6.9 Hz, 3H).
Chloride of (S) -2,2-Dimethyl-4-oxo-l, 3-dioxolane-5-acetyl. To 20 g (1149 mmol) of (S) -2, 2-Dimethyl-4-oxo-l, 3-dioxolane-5-acetic acid in a 250 ml three-necked round bottom flask was added under argon at room temperature 75 g (630 mmol) of thionyl chloride and 2 drops of DMF. The reaction mixture was stirred until evolution of gaseous HCl (oil sparger, about 2 hours). The excess thionyl chloride was distilled in vacuo and the remaining minimum quantities were removed under reduced pressure (9 hours). The acid chloride (22.1 g) thus obtained was used without further purification in the next step: IR (Neto csir * -) 2998, 1793, 1751, 989, 958; X H N (CDClj) d 4.69 (dd, J> 3.6, 6.4 Hz, iH), 3.56 (dd, J = 3.6, 18.1 Hz, 1H, 3.36 (dd, J - 6.4, 18.1 Hz, 1H), 1.65 ( s, 3H), 1.58 (S, 3H).
(S) -2,2-D-methyl-4-oxo-l, 3-dioxolane-5-acetaldehyde. To a 250 ml three-necked round bottom flask equipped with a mechanical stirrer, reflux condenser and gas inlet dispersion tube, 22.1 g (115 mmol) of (S) -2 chloride was added, Crude 2-dimethyl-4-oxo-l, 3-dioxolane-5-acetyl dissolved in 250 ml of anhydrous xylenes. To this solution were added 2. 0 g of 5% palladium on barium sulfate and 0.2 ml of auinoline-sulfur poison solution (prepared by placing reflux 1 g of sulfur with 5 ml of quinoline for 6 hours and diluting to a final volume of 70 ml with anhydrous xylenes). Hydrogen gas was bubbled through the reaction mixture and the hydrochloric acid generated was trapped in 175 ml of water containing a few drops of phenolphthalein indicator. The mixture was heated to 135 ° C and monitored by titration of the hydrochloric acid solution with 5M sodium hydroxide solution. Upon completion (approximately 3 hours), the reaction mixture was cooled to room temperature and 1.5 g of Norit was added. The mixture was filtered (Celite) and the filtrate was concentrated under reduced pressure. The residue was purified on silica gel using ethyl acetate: hexanes (1: 3) to give 16.2 g (89%) of white solid: m.p. 37-38ßC; [a] D25 1.4 ° (c- 4.54, CHjOH); IR (Neto crn'1) 2994 2744, 1793, 1725, 1386; : H NMR (CDCl,) d 9.78 (s, 1H), 4.80 (dd, J-3.6, 6.8 Hz, 1H), 3.10 (dd, J «3.6, 18.3 Hz, 1H); 2.92 (dd, J »6.9, 18.3 Hz, 1H), 1.63 (s, 3H), 1.58 (s, 3H); HRMS calculated for C7H10O4 (NT) 158.0579, Encont.158.0572.
(S) -5- (3-Methoxyallyl) -2,2-dimethyl-l, 3-dioxolane-4-one. In a 250 ml three-neck round bottom flask under argon, 14.2 g (126.58 mmoles) of potassium t-butoxide were dissolved in 300 ml of anhydrous tetrahydrofuran. The solution was cooled to 0 ° C and 44 g (126.6 mmoles) of methoxymethyltriphenylphosphine chloride were added slowly and with stirring (20 minutes). The resulting orange-red solution was stirred at 0 ° C for 45 minutes and 10 g (63.3 mmol) of (S) -2,2-Dimethyl-4-oxo-1,3-dioxolane-5-acetaldehyde in 50 ml of Anhydrous tetrahydrofuran was added dropwise (15 minutes). The mixture was allowed to stir at room temperature for one hour and was quenched by addition of 100 ml of brine. After being stirred for one hour, the mixture was extracted with 3 x 250 ml of ether. The combined ether extract was washed with 2 x. 150 ml of brine (Na2 =? 4) and filtered. The filtrate was evaporated under reduced pressure to give 18 g of a crude brown liquid (contaminated with triphenylphosphorane). The residue was purified on silica gel using ethyl acetate: petroleum ether (1: 9) to give 9.2 g (78%) of a mixture of enamel ethers E: Z inseparable as a colorless liquid [c.] D2S -3.2 ° (c = 2.8, CH "OH); IR (neat, cm "1) 2994, 2938, 1793, 1658;? NMR (CDCl,) d for the E-enol ether (75%) 6.42 (d, J-12.7 Kz, 1H), 4.49-4.34 (m , 2H), 3.53 (s 3H), 2.72-2.32 (m, 2H), 1.60 (S, 3H), 1.54 (S, 3H), For the ether E-enolic (25%) 6.04 (d, J - 6.1 Hz, 1H), 4.78-4.61 (m, 2HJ, j.bl (s, 3H ), 2.72-2.32 (m, 2H), 1.60 (s, 3H), 1.54 (s, 3H); HRMS calculated for C, Hl404 (M *) 186.0892, Encont.186.0894. (2S) -Methyl tetrahydro-5-methoxy-2-furoate. To enol ethers, (S) -5- (3-methoxyallyl) -2, 2-dimethyl-1,3-dioxolane-4-one (4.0 g, 21.5 mmol), dissolved in 150 ml of anhydrous methanol, were added 5-6 drops of concentrated H2SO4. The resulting solution was heated for 6 hours and cooled to room temperature. Sodium bicarbonate (0.5 g) was added, and the methanol was removed in vacuo. The residue was dissolved in 250 ml of CH 2 Cl 2 and washed with 2 x 100 ml of saturated sodium bicarbonate solution and 2 x 125 ml of brine. The organic extract was dried (Na 2 S 4) and the solvent was removed under reduced pressure to produce a colorless liquid which was purified on silica gel using ethyl acetate: hexanes (1: 1) as eluent to provide 2.93 g (86%) ) of a colorless liquid as a mixture of 3: 1 diastereomers: [aJD2S 27.3 ° (c-1.2, CH, OH); IR (Net cm "x) 2958, 1739, 1213, 1105; XH NMR (CDCl,) d for, diastereomer A ($ 6) 5 # 21 { P 1H) # 4,64.4.52 (ra, 1H), 3.77 (s, 3H), 2.44-1.83 (m, 4H), for diastereomer, B (33%) 5.08 (m, 1H), 4.64-4.52 (m, 1H), 3.77 (s, 3H), 3.42 (s) , 3H), 2.44-1.83 (, 4H); .HRMS calculated for C, Hu04 (M4) 160.0735, Encont.160.0718. (2S) -Methyl tetrahydro-5-hydroxy-2-furoate. (2S) -Tetrahydro Methyl 5-methoxy-2-furoate (2.93 g, 18.3 mmol) was stirred with 500 ml of 25% aqueous acetic acid for about 10 hours (monitored by CCD). Upon completion of the reaction, the aqueous acetic acid was stirring in vacuo and the residue was purified on silica gel using ethyl acetate: hexane (1: 1) to give 2.5 g (90%) of a colorless liquid as a diastereomeric mixture of 5.8: 4.2: [ft] ss 9 3ß (and = 2.7, CH, 0H); IR I Net aiT1) 3457 (br), 2956, 1735, 1062, 1010; -? NMR (CDCl,) 5 For diastereomer A (58%) 5.62 (m, 1H), 4.60 (dd, J = 6.5, 8.1 Hz, 1H), 3.78 (s, 3H), 2.46-1.93 (m, 4H); for diastereomer-B (42%) 5.75 (m, 1H) 4.73 (dd, J * 3.8, 8.5 Hz, 1H), 3.76, (s, 3H), 2.46-1.93 (m, 4H); HRMS calculated for CßH10O4 (M *) 146.0579, Found. 146.0574.
(S) -2-Methyl hydroxyrachidonate. To a round, three-necked, flame-dried flask equipped with a low temperature thermometer and a rubber partition, 12.35 g (22.57 mmoles) of 3, 6, 9-pentadecatrienotriphenylphosphine bromide and 350 were added under argon. ml of anhydrous tetrahydrofuran. The solution was cooled to -35 ° C and 14.1 ml of n-BuLi at 1.6 M in hexanes (22.56 mmoles) were added dropwise with stirring. The dark red solution was warmed to room temperature, stirred for an additional 30 minutes and cooled to -60 ° C, and (2S) -Tetrahydro-5-hydroxy-2-methyl furoate (1.65 g, 11.28 mmol) was dissolved in 25 ml of anhydrous tetrahydrofuran was added dropwise (approximately 15 minutes). The mixture was stirred at -60 ° C for 2 hours and warmed to room temperature. Upon completion (monitoring with CCD, about 8-9 hours), the reaction was quenched by the addition of 100 ml of 10% aqueous HCl and extracted with 3 x 300 ml of ethyl acetate. The organic layers were washed with 3 x 250 ml of water, 2 x 200 ml of brine, dried (Na 2 SO 4), filtered and concentrated in vacuo. The residue contaminated with triphenylphosphorane was purified on silica gel using ethyl acetate: hexanes (1: 5) as eluent to provide 3.1 g. (82%) yellow oil: [] D25 10.2 »(c = 5.4, CH, OH); IR (Net cm ---) 3477 (br), 3012, 2956, 1739, 1652, 721; lH RN (CDCl,) 5 5.45-5.25 (m, 8H), 4.20 (dd, J-4.0, 7.6 Hz, 1H), 3.79 (s, 3H), 2.92-2.74 (m, 4H), 2.35-1.92 ( m, 4H), 1.91-1.62 (m, 2H), 1.49-1.24 (m, 6H), 0.89 (t, J - 6.4 Hz, 3H); HRMS calculated for C "H3403 (M *) 334.2507, found 334.2509; analysis calculated for Cv, H -, _. 0,: C, 75.41; H, 10.25. Encont. C, 75.36; H, 10.13.
EXAMPLE B Chloride of (R) -2,2-Dimethyl-4-oxo-l, 3-dioxolane-5-acetyl was prepared as for the S-isomer of Example A of R-malic acid in a similar yield: IR, Ne or crn 1) 2996, 1793, 1751, 989 958; ** H NMR (CDCl,) d 4.69 (dd, J-3.6, 6.4 Hz, 1H), 3.53 (dd, J-3.6, 18.1 Hz, 1H) 3.35 (dd, J-6.4, 18.1 HZ, 1H) 1.65 (S, 3H), 1.58 (s, 3H).
(R) -2,2-Dimethyl-4-oxo-l, 3-dioxolane-5-acetaldehyde was prepared as for the S-isomer of (R) -2, 2-Dimethyl-4-oxoyl chloride , 3- dioxolane-5-acetyl in a similar yield: tpp 37-38 ° C; [a] D2S 3.7β (c = 5.12, CH, OH); IR (Net c "1) 2996, 2746, 1791, 1727, 1388; ** H NMR (CDCl,) d 9.78 (3, 1H), 4.80 (dd, J - 3.6, 6.8 Hz, 1H), 3.11 (dd) , J - 3.6, 18.3 Hz, 1H), 2.93 (dd, J - 6.9, 18.3 Hz, 1H), 1.63 (a, 3H), 1.58 (S, 3H); HRMS calculated for • C7H10O4 (M *) 158.0579, found.158.0574.
(R) -5- (3-Methoxyallyl) -2,2-dimethyl-4-oxo-l, 3-dioxolane-4-one was prepared as for the S-isomer of (R) -2,2-dimethyl-4-oxo -l, 3-dioxolane-5-acetaldehyde in a similar yield: ra? as 3.1 * (C-3.07, CH3OH); IR (Net aiT1) 2992, 2740, 1793, 1656; lH NMR (CDCl,) for thß E-enol ether (72%) d 6.42 (d, J- 12.7 Hz, 1H), 4.49-4.34 (, 2H), 3.53 (s, 3H), 2.72- 2.32 (ra, 2H), 1.61 (s, 3H), 1.54 (s, 3H); for thß Z-enol ßther (28%) 6.05 (d, J - 6.1 Hz, 1H), 4.78-4. &S (m, 2H), 3.61 (S, 3H), 2.72-2.32 (m, 2H) , 1.61 (S, 3H), 1.54 (S, 3H); HRMS calculated for C, H1404 (M *) 186.0892, Found. 186.0891. (2R) -Methyl tetrahydro-5-methoxy-2-furoate was prepared as for the S-isomer of (R) -5- (3-Methoxyallyl) -2,2-dimethyl-4-oxoyl, 3-dioxolane-4 -one in a similar yield: raj as -28.33 ° (c-0.1, CH, 0H); IR (Net cm'1) 2956, 1754, 1209, 1105; LH NMR (CDCl,) diastereomer A (66%) d 5.21 (m, 1H), 4.64-4.52 (m, 1H). 3.77 (s, 3H), 3.38 (s, 3H), 2.42- 1.84 (m, 4H) for diastereomer B (33%) 5.08 (m, 1H), 4.64- 4.52 (m, 1H), 3.77 (a, 3H) ), 3.42 (s, 3H), 2.42-1.84 (, 4H); HRMS calculated for - CH, ^ (M #) 160.0735, Encont. 160.0750. (2R) -Methyl tetrahydro-5-hydroxy-2-furoate was prepared as for the S isomer of methyl (2R) -tetrahydro-5-methoxy-2-furoate in a similar yield: [a] D2 $ -9.2 ° (c »1.8, CHjOH); IR (Net ap'1) 3543 (br), 2956, 1741, 1068, 1OX0; XH NMR (CDCLj) for diastereomer A (58%) d 5.62 (m, 1H), 4.67 (dd, J = 6.5, 8.1 Hz, 1H), 3.78 (s, 3H), 2.46-1.93 (m, 4H)? for diastereomer B (42%) d 5.75 (m, 1H), 4.73 (dd, J »3.8, 8.5 Hz, 1H), 3.76 (s, 3H), 2.46-1.93 (m, 4H); HRMS calculated for CßH10O4 (M *), 146.0579, Found. 146.0577.
(R) -2-Methyl hydroxyarachidonate was prepared as for the S isomer of methyl (2R) -tetrahydro-5-hydroxy-2-furoate in a similar yield: lr? A # j1o 2s - -1.0 n. 5 ee °, / (C- = "" 0.9?, ^ CHtjO "H".) "; - IR (, Net cm "1) 3504 (br), 3012, 2956, 1739, 1652, 723; XH NMR (CDCLj) d 5.45-5.25 (m, 8H), 4.20 <dd, J - 4.0, 7.7 Hz , 1H), 3.79 (s, 3H), 2.92-2.74 (m, 4H), 1.90-1.62 (m, 2H), 1.49-1.24 (m, 6H), 0.89 (t, J - 6.4 Hz, 3H), HRMS calculated for C ^ H ^ 0, (M *), 334.2507, Encont.334.2506.
EXAMPLE C Pentadecyl rifenyl phosphonium bromide. Under a nitrogen atmosphere, 1-bromopentadecane (6.5 g, 22.4 mmol) was treated with 5.87 g (22.4 mmol) of triphenylphosphine in 15 ml of acetonitrile and heated to 135 ° C. The reaction was monitored by CCD (salt formation, approximately 18 hours), and the mixture was dried under reduced pressure (24 hours) to ensure the removal of minimal amounts of acetonitrile and gave 12.3 g of a colorless solid which was used in the next reaction without further purification. (2S, 5Z) -2-hydroxy-5-eicosenoate methyl. To a flame-dried 500 ml 3-necked flask equipped with a low temperature thermometer and a rubber partition, 12.2 g 9 (22.06 mmoles) of pentadecylphosphonium bromide and 300 ml of anhydrous tetrahydrofuran were added under argon. After cooling to -35 ° C, 13.8 ml of n-BuLi were added dropwise to 1.6 M in hexanes (22.06 mmoles) and with stirring. The orange solution was warmed to room temperature and stirred for an additional 30 minutes. The mixture was cooled to -35 ° C, and 18.5 g (103.3 mmol) of hexamethylphenoramide was added slowly. The reaction mixture was stirred for 45 minutes and cooled to -60 ° C. (2S) -tetrahydro-5-hydroxy-2-methyl furoate (1.6 g, 11. 03 mmoles) was dissolved in 25 ml of anhydrous tetrahydrofuran and added dropwise, and the stirring was continued for 1 hour.
Time at -60 ° C. The mixture was warmed to room temperature, the conclusion of the reaction was monitored using CCD The reaction was quenched by the addition of 100 ml of 10% aqueous Hel's solution and extracted with 3 x. 300 ml of ethyl acetate The organic layers were washed with 3 x 250 ml of water, 2 x 200 ml of brine, dried (a2S? 4), filtered and evaporated in vacuo, the residue, contaminated with triphenylphosphorane, was purified on silica gel using ethyl acetate: hexanes (1: 5) to give 2.98 g (80%) of colorless oil:. [< ?) 03S +8.9 (C = s.l.4, CH3OH); IR Net a'1) 3475 (br), 3006, 2925, .1-739, 721; XH NMR (CDjCOCD,) d 5.48-5.24 (m, 2H), 4.20 (m, 1H), 3.79 (s, 3H), 2.72 (d, J * 5.3 Hz, 1H), 2.32-2.15 (m, 2H) , 2.15-1.98 (m, 2H), 1.92-1.64 (mf 2H), 1.48-1.21 (m, 24H), 0.88 (t, J = »6.3 Hz, 3H); UC NMR. (CD, COCD,) 5 175.7, 131.5, 127.9, 70.0, 52.4, 34.4, 31.9, 29.7 (5C), 29.6 (2C), 29.3 (3C), 27.2, 22.7 (2C), 14.0; HRMS calculated for C HMO, (M *) 340.2977, found 340.2977; analysis calculated for Ca? H4o03: C, 74.07; H, 11.84 .: Encont: C, 74.13; H, 11.91.
EXAMPLE D (2R, 5Z) -2-hydroxy-5-eicosenoate methyl was prepared as for the S-isomer from (2R-tetrahydro-5-hydroxy-2-methyl furoate in a similar yield: [d] ^ * - 8.8 ° (c «2.1, CH, OH); IR (Net cm-1) 3482 (br), 3006, 2925, 1739, 721; ** H NMR (CD3COCD3) d 5.48-5.24 (m, 2H), 4.20 (m, 1H), 3.79 (a, H), 2.72 (d, J - 5.3 Hz, 1H), 2.32-2.15 (m, 2H), 2.15-1.98 (, 2H), 1.92-1.64 (m, 2H) , 1.48-1.21 (m, 24H), 0.88 (t, J = 6.3 Hz, 3H), 13C NMR (CD, COCD3) d 175.7, 131.5, 127.9, 70.0, 52.3, 34.4, 31.9, 29.7 (5C), 29.6 (2C), 29.3 (3C), 27.2, 22.7 (2C), 14.0; HRMS calculated for Ca? .H40O, (M *) 340.2977, Encopt.340.2970.
EXAMPLE 1 (S) -2- (Allyloxy) acetyloxy-methyl arachidinoate. To a dry two-necked round-bottomed flask equipped with a rubber partition was added, under argon, 1.5 g (4.49 mmol) of methyl (S) -2-hydroxy-arachidonate dissolved in 125 ml of anhydrous CH 2 Cl 2. The solution was cooled to 10 ° C (ice bath) and 1.3 g of allyloxyacetic acid (11.23 mmole) dissolved in 15 ml of anhydrous CH2CI2 and 0.133 g (0.90 mmole) of 4-pyrrolidinopyridine dissolved in 2 ml of CH2Cl2 was added. A solution of 2.32 g (11.23 mmoles) of DCC in 25 ml of CH2Cl2 was added dropwise with stirring, warmed to room temperature and stirred overnight. The CH2Cl2 was removed by distillation under reduced pressure and the residue was chromatographed on silica gel using ethyl acetate: hexane (1: 5) as eluent to give 1.79 g (92%) of yellow oil: .er] D--, -8.1β (c-0.1, CH 3 OH); IR (Net cpf x) 3012, 2956, 1751, 1652; ** H NMR (CDC13) d 5.94-5.82 (m, 1H), 5.41-5.22 (m, 10H), 5.11 (t, J - 6.3 Hz, 1H), 4.23 (d, J - 16.6 Hz, 1H), 4.22 (d, J - 16.6 Hz, 1H), 4.12 (d, J • = 1.3, 5.7 Hz, 2H), 3.75 (s, 3H), 2.89-2.68 (m, 4H), 2.32-1.89 (m.2H ), 1.42-1.31 (m, 6H), 0.89 (t, 6.5 Hz, 3H); HRMS calculated for C ^ H ^ Oj (M *) 432.2875, Encont. 432.2856; analysis calculated for C ^ HITA: C, 72.19; H, 9.32: Encont. C, 71.90; H, 9.11.
(S) -3- (Allyloxy) -4-hydroxy-5- [all-Z) -3,6, 9, 12, octadecatetraenyl] -2 (5H) -furanone. To a 250 ml three-neck round bottom flask dried with argon flame equipped with a low temperature thermometer and a partition was added 1.23 g (7.59 mmoles) of '- "hexamethyldisilazane in 100 ml of anhydrous tetrahydrofuran. The contents were cooled to -25 ° C (dry ice, CCI4) and 4.75 ml of n-BuLi at 1.6 M (7.59 mmoles) in hexanes was added dropwise with stirring while maintaining the temperature below 15 ° C. The stirred reaction mixture was heated to -5 ° C, the contents were maintained between -5 ° C and 0 ° C for 45 minutes, and cooled to -78 ° C (dry ice / acetone). (S) -2- Methyl hydroxyrachidonate (allyloxy) acetate (1.56 g, 3.61 mmol) in 30 ml of anhydrous tetrahydrofuran was added dropwise with stirring while maintaining the temperature below -68 ° C. After the addition, the mixture was stirred at -68 ° C for 75 minutes and cooled by the addition of 40 ml of 10% aqueous HCl solution. Ether (125 ml) was added, the mixture was warmed to room temperature and 3 x 10 ml of ether was extracted. The ether extract was washed with 2 x 75 ml of brine, dried (a2S? 4) and concentrated in vacuo to yield 1.34 g of crude product which was purified on silica gel using 10% methanol in chloroform as eluent to provide 1.28 g (89%) yellow oil:. .2 S -9.7 ° (c = 0.2, CH, 0H); IR (Net OtL'x) 3081 (brj, 3012, 2956, 1747, 1670, 723; lH MN (CD, C0CD3) d 6.05 ^ 5.89 (m, 1H), 5.44-5.14 (m, 10H), 4.72 ( dd, J - 3.5, 7.7 Hz, 1H), 4.48 (dt, J «1.2, 5.6 Hz, 2H), 2.89-2.69 (m, 6H), 2.38- 1.88 (m, 4H), 1.71-1.21 (, 8H ), 0.87 (t, J »6.6 Hz, 3H); MS (FAB) (M + l) * 401; HRMS calculated for C, ßH3í04 M * 400.2614), found 400.2606; analysis calculated for C "H, ß04 + HaO: C, 71.74; H, 9.15: Encont; C, 71.90; H, 9.11.
(S) -4-hydroxy-5- [all-Z) -3,6,9,18-octadecatetraenyl] -3- [(E) -propynyloxy] -2 (5H) -furanone. To a 250 ml three-necked round bottom flask, flame-dried, 0.254 g (0.30 mmol) of [bis (methyldiphenylphosphino)] (1,5-cyclooctadiene) iridium (1) hexafluorophosphate suspended in argon was added under argon. 50 ml of anhydrous tetrahydrofuran without freshly distilled peroxide. The flask was evacuated and the argon was displaced with nitrogen. The red suspension turned into a colorless solution and after 5 minutes the flask was evacuated and replaced with argon. The (S) -3- (allyloxy) -4-hydroxy-5- [all-Z) -3,6,9,16 octadecatetraenyl] -2 (5H) -furanone (0.6 g, 1.5 mmol), dissolved in 25 ml of tetrahydrofuran without peroxide was added and the conclusion of the reaction was monitored using CCD (approximately 3 hours). The solvent was evaporated under reduced pressure and the residue was purified on silica gel using 10% methanol in chloroform as eluent to give 0.47 g. (79%) of a dark yellow oil: 0.2, CH, 0H); IR (Net cm * 1) 3081 (br), 3012, 2956, 1745, 16ß2, 721; XH MN (CD, C0CDj) d 6.42-6.35 (m, 1H), 5.49- 5.23 (m, 8H), 5.05-4.89 (, 1H), 4.77 (dd, J - 3.5, 7.9 Hz, 1H), 2.92- 2.74 (m, 4H), 2.31-1.57 (m, 6H), 1.S1 (dd, J - 1.6, 6.9 Hz, 3H), 1.45-1.28 (m, 8H), 0.87 (t, J - 6.4 Hz, 3H); MS (FAB) (M + 1) * 401, (M + Na) + 423; HRMS. calculated for C2 $ H, §04 (M *) 400.2614, Encont, 400.2616; analysis calculated for C, 71.74; H, 9.15: EnconU: C, 71. 70; H, 9.06.
(S) -3,4-dihydroxy-5- [odos-Z) -3,6,9,12-octadecatetraenyl] -2 (5H) furanone. To a 100 ml round bottom flask was added, under nitrogen, 0.3 g (0.74 mmoles) of (S) -4-hydroxy-5 [(All-Z) -3,6,9, 12-octadecatetraenyl] -3 - [(E-propenyloxy] -2 (5H) -furanone dissolved in 60 ml of 50% aqueous acetic acid The stirred solution was heated to reflux (oil bath) for 15 minutes, cooled, and concentrated in vacuo. The residue was chromatographed on silica gel using 12% methanol in chloroform as eluent to give 0.26 g (95%) of yellow oil: [a] D2s .13? 5ß (c-0.2, CHjOH); IR (Net, cm "1) 3220 (br), 3012, 2956, 1751, 1670, 1652, 723, 694; XK NMR (CD, COCD3) d 5.48-5.34 (ra, 8H), 4.68 (dd, J» 3.4, 7.9 Hz , 1H), 2.85-2.72 (m, 4H), 2.25-2.19 (, 2H), 2.18-1.97 (, 4H), 1.65-1.52 (m, 1H), 1.48-1.28 (m, 7H), 0.87 (t , J - 6.7 Hz, 3H); HRMS calculated for C22H3204 (M *) 360.2301, Encont 360.2308, analysis calculated for C "H" 04 + 0.33 H20: C, 72.1; H, 8.98: Found: C, 72.18; H, 8.93.
EXAMPLE 2 (R) -2- (2-allyloxy) methyl acetyloxy-arachidonate was prepared for the S-isomer from methyl (R) -2-hydroxy-arachidonate in a similar yield: faj 23 7 g0, 0.1, CH, OH); IR (Net, era'4) 3012, 2956, 1756, 1648; * H NMR (CDCl,) 6 5.94-5.82 (Bf 2H), 5.45-5.22 (m, 10H), 5.11 (t = 6.3 KZ, IH); 4.23 (d, J _ .16 (6 Hz, 1H), 4.22 (d, J, '16-6, 1H), 4.12 (dt, J =! .3, 5.7 Hz, 2H) / 3 ^ ^ ^ 2.89 -2.68 (m, 4H), 2.28-1.89 («, SH), 1.61-1.49 (rp, 2H) '1.42-1.31 (, 6H), 0.89 (t, J = 6.5 Hz, 3H); HRMS calculated for C "H40O5 (M *), 432.2875, Encont 432.2858.
(R) -3- (Allyloxy) -4-hydroxy-5- [(all-Z) -3,6,9,1-octadecatetraenyl] -2 (5H) -furanone was prepared as for the S-isomer from (R) -2-methyl hydroxy-qurachidonate, (allyloxy) acetate in a similar yield: [or *] DM 9.4 ° (c »0.3, CH30H); IR (Net crn * 1) 3081 (br), 3012, 2956, 1749, 1670, 723; * H NMR (CD, COCD,) d 6.05-5.89 (m, 1H), 5.44-5.14 (m, 10H), 4.74 (dd, J »3.5, 7.7 Hz, 1H), 4.48 (dt, J - 1.1, 5.7 Hz, 2H), 2.87-2.69 (m, 6H), 2.42-1.88 (m, 4H), 1.71-1.21 (m, 8H), 0.87 (t, J - 6.6 Hz, 3H); MS (FAB) (M + D * 401; HRMS calculated for C "H3t04 (M *) 400.2614, Found 400.2607.
(R) -4-hydroxy-5- [(all-Z) -3,6,9,1-octadecatetraenyl] -3- [(E) -propenyloxy] -2 (5H) -furanone was prepared as for the isomer S from (R) -3- (allyloxy) -4-hydroxy-5- [(all-Z) -3, 6, 9, 12-octadecatetraenyl] -2 (5H) -furanone in a similar yield: [ aiuV * 11.70 (c-0.2, CH3OH); IR (Net, cm "1) 3081 (br), 3012, 2956, 1749, 1664, 696; * HRN (CD3COCD3) d 6.42-6.35 (m, 1H), 5.49-5.23 (m, 8H), 5.05- 4.89 (m, 1H) 4.77 (dd, J = 3.5, 7.9 HZ, 1H), 2.92-2.74 (m, 4H), 2.31- 1.57 (m, 6H), 1.51 (dd, J * 1.6, 6.9 Hz, 3H ), 1.45-1.28 (m, 8H), 0.87 (t, J »6.4 Hz, 3H); MS (FAB) (M + l) * 401, (M + Na) * 423; HRMS calculated for C2 $ H3s04 ( M *) 400.2614, Encont 400.2615.
(R) -3,4-dihydroxy-5- [(all-Z) -3,6,9,1-octadecatetraenyl] -2 (5H) -furanone was prepared as for the S-isomer from (R) - 4-hydroxy-5- [(all-Z) -3,6, 9,12-octadecatetraenyl] -3- [(E) -propenyloxy] -2 (5H) -furanone in a similar yield: rßj as. ^ ^ or id-0.2, CH 3 OH); IR (Net, crn "1) 3241 (br), 3012, 2956, 1751, 1675, 1652, 723, 694; ** H NMR (CD3COCD3) d 5.48-5.34 (m, 8H), 4.68 (dd, J - 3.4, 7.9 Hz, 1H), 2.85-2.72 (m, 4H), 2.35-2.19 (ra, 2H), 2.18-1.97 (m, 4H), 1.65-1.52 (ra, 1H), 1.48-1.28 (m, 7H), 0.87 (t, J * 6.7 Hz, 3H), HRMS calculated for CJ3HJ204 (M *) 360.2301, Encorit 360.2305, analysis calculated for C "H3a04 + 0.5 H20: C, 71.51; H, 9.00; 71.52; H, 9.14.
EXAMPLE 3 (2S, 5Z) -2- (2-Allyloxy) acetyloxy-5-eicosenoate methyl. To a three-necked, flame-dried round-bottomed flask equipped with a rubber partition, 0.65 g (1.91 mmol) of methyl (2S, 5Z) -2-hydroxy-5-eicosenoate was added under argon and 50 ral of anhydrous CH2CI2. After cooling to 10 ° C (ice bath), 0.45 g (3.82 mmoles) of allyloxyacetic acid dissolved in 15 ml of anhydrous CH2Cl2 and 0.028 g (1.29 mmoles) of 4-pyrrolidinopyridine dissolved in 2 ml of water were added. CH2Cl2 anhydrous. A solution containing 0.79 g (3.82 mmoles) of DDC in 15 ml of CH2Cl2 was added dropwise and the stirred mixture was warmed to room temperature and stirred for an additional 8 hours. CH2Cl2 was evaporated in vacuo and the product was purified on silica gel using ethyl acetate: hexanes (1: 5) as eluent to give 0.78 g (93%) of colorless oil: 1.6, CH3OH); IR (Net.cm * 1) 3006, 2923, 1758, 721; XH NMR (CD3COCD3) 5 6.01-5.86 (m, 1H), 5.51-5.24 (m, 4H), 5.10 (t, J - 6.3 Hz, 1H), 4.22 (d, J - 16.6 Hz, 1H), 4.23 (d, J = 16.6 Hz, 1H), 4.13 (dt, J • 1.1, 5.7 Hz, .2H), 3.75 (S, 3H), 2.26-2.12 (m, 2H), 2.07-1.87 (m, 4H ), 1.42-1.28 (m, 24H), 0.88 (t, J - 6.3 Hz, 3H); HRMS calculated for (M), 438.3345 Encont438.3330.
(S) -3-Allyloxy-4-hydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone. To a 250 ml round, three-neck, flame-dried flask equipped with a low temperature thermometer and a rubber partition, 0.648 g (4.02 mmol) of hexamethyldisidalazane in 50 ml of anhydrous tetrahydrofuran was added under argon. The contents were cooled to -25 ° C) dry ice / CCl), and 2.51 ml of n-BuLi at 1.6 M (4.02 mmoles) in hexanes were added dropwise with stirring while maintaining the temperature below -15 ° C. The mixture was heated to -5 ° C and the contents were maintained between -5 ° C and 0 ° C for 45 minutes. The mixture was cooled to -78 ° C (dry ice / acetone) and 0.84 g (1.91 mmol) of (2S, 5Z) -2- (2-Allyloxy) acetyloxy-5-eicosenoate in 20 ml of anhydrous tetrahydrofuran was added dropwise with stirring while maintaining the temperature below -78 ° C for 75 minutes and extinguished with 10 ml of 10% aqueous HCl solution. Ether (75 ml) was added and the reaction mixture was warmed to room temperature and extracted with 3 x. 75 ml of ether. The ether extracts were washed with 2 x 50 ml of brine, dried (a2SO4) and concentrated in vacuo to provide 0.75 g (97%) of residue. Purification on silica gel using 10% methanol in chloroform as eluent gave 0. 69 g (89%) of white solid: m.p. 51-54 ° C; [a30"-9.0 ° (C-0.6, CHjOH); IR (Net cm" 1) 3079 (br), 3002, 2915, 1741, 1654, 734, 719; 4H NMR (CD3C0CD.}.) D 6.04-5.89 (m, 1H), 5.47-5.15 (m, 4H), 4.72 (dd, J-3.5, 7.6 Hz, 1H), 4.48 (dt, J-1.1, 5.9 Hz, 2H), 2.21-1.54 (m, 6H). 1.45-1.28 (m, 24H), 0.87 (t, J - 6.8 Hz, 3H); HRMS calculated for C3SH4204 (M *) 406.3083, Encont 406.3084; analysis calculated for CjjH ^ O *: C, 73.85; H, 10.41. Encont. C, 73.52; H, 10.27. (a) -4-hydroxy-5- [(Z) -3-octadecenyl] -3- [(E) -propenyloxy] -2 (5H) -furanone. To a 250 ml three-necked round bottom flask, flame dried, under argon, 0.172 g (0.20 mmol) of [bis (methyldiphenylphosphine] (1,5-cyclooctadian) iridium (I) hexafluorophosphate suspended in 35 ml was added. of anhydrous tetrahydrofuran without freshly distilled peroxide The flask was evacuated and the argon was displaced by hydrogen The red suspension became a colorless solution after 5 minutes The flask was evacuated and the hydrogen replaced by argon (S) 3-Allyloxy-4-hydroxy-5- [(Z) -3- octadecenyl] -2 (5H) -furanone (0.415 g, 1.02 mmol) was dissolved in 25 ml of tetrahydrofuran without peroxide and added to the activated catalyst. Upon completion (CCD, about 3 hours) the solvent was evaporated in vacuo and purified on silica gel using 10% methanol in chloroform as eluent to provide 0.33 g (79%) of a white waxy solid: mp 42-45 ° C; [a) 0 »-13.0 ° (C = 0.3, CHsOH); IR (Net, era'1) 3079 (br), 3004, 2919, 1739, 1681, 1658, 738, 721; XH NMR (CD3COCD3 d 6.43-6.36 (m, 1H), 5.49-5.31 (m, 2H), 5.05-4.91 (m, 1H), "4.77 (dd, J» 3.5, 7.8 Hz, 1H), 2.24-1.62 (m, 6H), 1.51 * "(dd, 'J' 1.7, 6.9 Hz, 3H), 1.44-1.24 (m, 24H), 0.87 (t, J» 6.8 Hz, 3H); HRMS calculated for C ^ jO , (M *) 406.3083, Encont 406.3083.
(S) -3,4-Dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone. To a 100 ml round bottom flask was added, nitrogen garlic, 0.2 g (0.49 mmole) (S) -4-hydroxy-5- [(Z) -3-octadecenyl] -3- [(E) -propenyloxy] -2 (5H) -furanone was dissolved in 60 ml of 50% aqueous acetic acid. The solution was heated to reflux (oil bath) for 15 minutes, cooled and the aqueous acetic acid was removed in vacuo. The residue was purified on silica gel using 12% methanol in chloroform as eluent to give 0.17 g (95%) of a white waxy solid: m.p. 64-66 ° C; [a] D "-9.9 ° (e =» 0.5, CH, 0H), IR (Net CUT1) 3421 (br), 2917, 2850, 1754, 1668, 719; lH NMR (DsCOCD3) d 5.49-5.28 (m , 2H), 4.67 (dd, J - 3.4, 7.8 Hz, 1H), 2.32- 1.88 (m, 4H), 1.69-1.51 (m, 2H), 1.44-1.25 (m, 24H), 0.87 (t, J * 6.7 HZ, 3H); HRMS calculated for C22H3804 (M *) 366.2770, Encont 366.2780, analysis calculated for C22H3g04 + 0.33 H20: C, 70.93; H, 10.46; Found: C, 70.73; HL.10.31.
EXAMPLE 4 (2r, 5z) -2- (2-allyloxy) acetyloxy-5-eicosenoate methyl was prepared as the S-isomer from methyl (2R, 5Z) -2-hydroxy-5-eicosenoate (as prepared in Example D) in a similar yield: toe] 02 * 6.4 ° (s = 1.4, CH3OH); IR (Net, was "1) 3006, 2921, 1756, 721; ** H NMR (CDsC0CD3) d 6.01-5.86 (m, - '1H), 5.51-5.24 (ra, 4H), 5.10 (t , J - 6.3 Hz, 1H), 4.22 (d, J - 16.6 Hz, 1H), 4.23 (d, J - 16.6 Hz, 1H), 4.13 (dt, J - 1.1, 5.7 Hz, 2H), 3.75 (s) , 3H), 2.26-2.12 (m, 2H), 2.07-1.87 (mr 4H), 1.42-1.28 (m, 24H), 0.88 (t, J * 6.3 Hz, 3H); HRMS calculated for C ^ H ^ Oj (M *) 438.3345, Encont438.3347.
(R) -3-Allyloxy-4-hydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone was prepared as for the S-isomer from (2R, 5Z) -2- (2 - allyloxy) acetyloxy-5-eicosenoate methyl in a similar yield: mp 49-53 ° C; [a] 0 «8 > 9o (c to 0 8 / cH, OH); IR (Net cm. 3079 (br), 3002, 2915, 1741, 1654, 734, 719; XH NMR (CDjCOCDj) 6.04-5.89 (m, 1H), 5.47-5.15 (m, 4H), 4.72 (dd, J »3.5, 7.6 Hz, 1H), 4.48 (dt, J - 1.1, 5.9 Hz, 2H) , 2.21-1.54 (m, 6H), 1.45-1.28 (ra, 24H), 0.87 (t, J »6.8 Hz, 3H); HRMS calculated for C2SH4204 (M *), 406.3083 Encont 406.3094.
(R) -4-hydroxy-5- [(Z) -3-octadecenyl] -3- [(E) -propenyloxy] -2 (5H) -furanone was prepared as for the S-isomer from (R) - 3- allyloxy-4-hydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone in a similar yield: mp 42-45 ° C; [< ?] B3S 12.8 ° (c »0.3, CH3OH); IR (neat, cm'1) 3079 (br), 3004, 2919, 1739, 1681, 1658, 738, 721; lH NMR (CD, C0CD3) d 6.43-6.36 (ra, 1H), 5.49-5.31 (m, 2H), 5.05-4.91 (ra, 1H), 4.77 (dd, J * 3.5, 7.8 Hz, 1H), 2.24 -1.62 (m, 6H), 1.51 (dd, J-1.7, 6.9 • ** HZ, 3H), 1.44-1.24 (m, 24H), 0.87 (t, J = 6.8 Hz, 3H); HRMS calculated for Cj ^ A (M *) 406.3083, Encont: 406, 3076.
(R) -3,4-Dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone was prepared as for the S-isomer from (R) -4-hydroxy-5- [( Z) -3-octadecenyl] -3- [(E) -propenyloxy] -2 (5H) -furanone in a similar yield: mp 65-67 ° C; [] c 25 9.5 ° (C = 0.5, CH 3 OH); IR (Net cm "1) 3421 (br), 2917, 2850, 1754, 1666, 734, 719; XH NMR (CD3COCD3) d 5.49-5.28 (ra, 2H), 4.67 (dd.J = 3.4, 7.8 Hz, 1H), 2.32-1.88 (tn, 4H), 1.69-1.51 (m, 2H), 1.44-1.25 (m, 24H), 0.87 (t, J * 6.7 Hz, 3H); HRMS calculated, for C22H3.04 ( M *), 366.2770 Encont.366.2780, analysis calculated for • C23H3í04 + 0.33 H20: C, 70.93; H, 10.46: Encont: C, 70.53; H, 10.25.
EXAMPLE 5 (2S-5Z) -2- (2-benzyloxy) acetyloxy-5-eicosenoate methyl. To a two-neck, flame-dried, 100 ml round bottom flask, 0.275 g (0.81 mmol) of methyl (2S, 5Z) -2-hydroxy-5-eicosenoate in 40 ml of anhydrous CH2Cl2 was added under argon and 0.20 g (1.09 mmol) of benzine-acetyl chloride. The solution was cooled to 0 ° C (ice-salt bath), and pyridine (0-086 g, 1.09 mmol) was added dropwise. The mixture was stirred for 30 minutes at 0 ° C, warmed to room temperature and stirred for an additional 8 hours. The reaction was quenched with 10 ml of ice water. CH2Cl2 (20 ml) was added, and the mixture was stirred for 6 hours. The CH2Cl2 layer was washed with 3 x 20 ml of a 10% aqueous Hel's solution, 3 x 15 ml of saturated sodium bicarbonate solution, 2 x 25 ml of brine, dried (Na2 =? 4) and concentrated under reduced pressure. The residue was purified on silica gel using ethyl acetate: hexanes (1: 5) as an eluent to give 0.34 g (87%) of a white solid: [Qf] s2s-S.4 »(C * 0.5, CH3OH ); IR (Neto sra-i) 3006 (br), 2923, 1756, 1455, 734, 698; XH NMR (CD3COCD3) d 7.40-7.26 (m, 5H), 5.48-5.22 (rn, 2H), 5.12 (t, J * 16.7 Hz, 1H); 4.17 (d, J = 16.7 Hz, 1H), 3.76 (s, 3H), 2.20-1.87 (ra, 6H), 1.49-1.21 m, 24H), 0.88 (t, J - 6.4 Hz, 3H); HRMS calculated for C30H4ßOe (M *), 488.3501, Encont.488.3501.
(S) -3-benzyloxy-4-hydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
To a flame-dried three-neck 100 ml flask equipped with a low temperature thermometer was added under argon0.302 g (1.87 mmol) of hexamethyldisilazide in 25 ml of anhydrous tetrahydrofuran. The solution was cooled to -25 ° C and 1.17 ml of n-BuLi at 1.6 M (1.87 mmol) in hexanes was added dropwise with stirring while maintaining the temperature below -15 ° C. The reaction was maintained between -3 ° C and -5 ° C for 45 minutes, cooled to -78 ° C and 0.434 g (0.89 mmoles) of methyl (2S, 5Z) -2-hydroxy-5-eicosenoate, ( benzyloxy) acetate in 8 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stirred for 2 hours and quenched at -78 ° C with 10 ml of cooled 10% Hel aqueous solution. Ether (15 ml) was added and the mixture was warmed to room temperature and extracted with 3 x 50 ml of ether. The organic layers were washed with 3 x 25 ml of brine, dried (Na2 =? 4) and concentrated under reduced pressure. The residue was purified on silica gel using 10% methanol in chloroform as eluent to give 0.34 g (84%) of a white solid: m.p. 76-79 ° C; [af] D2s -13.8 ° (c = 0.3, CH3OH); IR (KBr, cm "1) 3033 (br), 3004, 2917, 1739, 1660, 1402, 1342, 738, 728, 696; 4H NMR (CD3COCD3) d 7.43-7.29 (m, 5H), 5.48-5 27 (m, 2H>, 5.06 (d, J - 11.3 Hz, 1H), 5.01 (d, J - 11.3 Hz, 1H), 4.69 (dd, J - 3.5, 7.6 Hz, 1H), 2.28-1.51 (m , 6H), 1.42-1.27 (m, 24H), 0.86 (t, J - 6.7 Hz, 3H); HRMS calculated for (^ o, (M *), 456.3239; analysis calculated £ "Ci '^ 4 + 0.2 H20; C, 75.68; H, 9.72; Found: C, '75 .57-H, 9.68.
(S) -3,4-Dihydroxy-5-octadecyl-2 (5H) -furanone. To a 250 ml hydrogenation bottle was added 0.02 g of 10% palladium on carbon in 5 ml of methanol. To this suspension was added 0.1 g (0.219 mmoles) of (S) -3-benzyloxy-4-hydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone dissolved in 15 ml of methanol. The hydrogenation was started at 2.81 kg / cm2 at room temperature. The reaction was monitored to complete by CCD (approximately 5 to 6 hours), filtered (Celite) and evaporated under reduced pressure. The residue was purified on silica gel using 10% methanol in chloroform as eluent to generate 58 mg (72%) of a white solid: m.p. 110-112 ° C; 112-C, W -6.8- (c-0.3, CH3OH); IR (Kbr, cm'4) 3380 (br), 2917, 2848, 1741, 1668; ? NMR (CD3COCD3) d 4.67 (dd, J 3.5, 7.3 Hz, 1H), 2.01-1.88 (m, 1H), 1.57-1.47 (, 1H), 1.42-1.23 (ra, 322), 0.87 (t, J - 6.8 H, 3H); HRMS calculated for c ^ 0, UH, 368.2927, Encont, 368 2928 • analysis calculated for C2aH40O4 + 0.5 H.O: C, 69.99; H, 10.95; Encont. C, 70.27; H, 11.22.
EXAMPLE 6 (2R, 5Z) -2- (2-benzyloxy) acetyloxy-5-eicosenoate methyl was prepared as the S-isomer from methyl (2R, 5Z) -2-hydroxy-5-eicosenoate in a similar yield: [ a] D? 5 5.2 ° (c * 0.5, CHjOH); GO . { Net crn "1) 3006, 2923, 1756, 1455, 734, 698; ** H NMR (CD3COCD3) d 7.40-7.26 (m, 5H), 5.48-5.22 (m, 2H), 5.12 (t, J» 6.4 Hz, 1H), 4.66 (s, 2H), 4.25 (d, J = 16.7 Hz, 1H), 4.17 (d, J »16.7 Hz, 1H), 3.76 (s, 3H), 2.20-1.87 (ra, SH ), 1.49-1.21 (m, 24H), 0.88 (t, J = 6.4 Hz, 3H); HRMS calculated for Cj0H48O5 (M *), 488.3501, Find 488.3501.
(R) -3-benzyloxy-4-hydroxy-5- t (Z) -3-octadecenyl] -2 (5H) -furanone was prepared for the S-isomer from (2R, 5Z) -2- (2- benzyloxy) acetyloxy-5-eicosenoate in a similar yield; p.f. 76-79 ° C [a] n 13.5"(c - 0.5, CH30H); IR (K * > r ' Ctn'1) 3033 (br), 3004, 2917, 1739, 1660, 1400, 1342, 738, 730; 696; ** H NMR (CD3COCD3) d 7.43-7.29 (m, 5H), 5.48-5.27 (m, 2H), 5.06 (d, J »11.3 Hz, 1H); 5.01 (d, J = 11.3 Hz, 1H); 4.67 (dd, J * 3.5, 7.6"Hz, 1H), 2.28-1.51 (m, 6H), 1.42-1.27 (ra, 24H), 0.86 (t, J« 6.7 Hz, 3H); HRMS calculated for CítH4404 ( M '), 456.3239, Encont.456.3243.
(R) -3,4-Dihydroxy-5- [3-octadecyl] -2 (5H) -furanone was prepared as for the S-isomer from (R) -3-benzyloxy-4-hydroxy-5- [( Z) -3-octadecenyl] -2 (5H) -furanone in a similar yield: mp 114-117 ° C; [ct] Bls 5.2 ° (c = 0.2, CHjOH); IR (Kbr, was "1) 3411 (br), 2917, 2848, 1754, 1668; lH NMR (CD3COCD3) 4.67 (dd, J = 3.5, 7.3 Hz, 1H), 2.01-1.38 (m, 1H > 1.57-1.47 (m, 1H), 1.42-1.23 (ra, 32H), 0.87 (t, J «6.8 Hz, 3H), HRMS calculated for C2aH400 (M *), 368.2927, Find 368.2927, analysis calculated for C2jH40O4 + 0.5 H20: C, 69.99; H, 10.95; Encont.C: 69.71; H, 11.09.
EXAMPLE 7 (S) -3-methyl f-lactanylate. In a round-bottomed flask equipped with a reflux condenser, 3.0 g (18.07 mmol) of S-phenylacetic acid in 225 ml of methanol containing 4 drops of concentrated sulfuric acid was heated to reflux for 7 hours. The reaction mixture was cooled, 0.6 g of sodium bicarbonate were added, and the methanol was evaporated under reduced pressure. The residue was taken up in 200 ml of ether, and the ether layer was washed with 2 x 75 ml of water, 2 x 100 ml of saturated sodium bicarbonate solution and 2 x 75 ml of brine, dried (Na 2 S 4) and evaporated under reduced pressure to provide 3.12 g of crude product which was purified on silica gel using ethyl acetate: hexanes (1: 5) 'to give 2.92 g (90%) of a crystalline solid. white: mp 46-47 ° C, IR (Kbr, ero-1) 3473 (br), 3029, 2954, 1739, 1496, 1454; XH NMR (CDC13) 7.34-7.19 (m, 5H), 4.50-4.43 ( ra, 1H), 3.77 (s, 3H), 3.14 (dd, J * 4.4, 13.9 Hz, 1H), 2.96 (dd, J - 6.8, 13.9 HZ, 1H), 2.72 (d, J - 6.2 Hz, 1H ); HRMS salculatßd for CX0H12O3 (M *), 180.0786, Found.iso .0786.
(S) -2- (2-benzyloxy) acetyloxy-3-phenylactate methyl. To a 250 ml two-necked round-bottomed flask with flame drying, under argon, 2.92 g (16.2 mmol) of methyl (S) -3-phenylactate in 80 ml of anhydrous CH2Cl2 was added. Benzyloxycetyl chloride (4.49 g, 24.33 mmol) was added with stirring and the solution was cooled to 0 ° C in an ice-salt bath. Pyridine (1.93 g, 24.33 mmol) was added dropwise and the contents of the reaction were stirred for 30 minutes at 0 ° C, warmed to room temperature and stirred for an additional 3 hours. The reaction was quenched with 40 mL of ice water and 75 mL of CH2Cl2 was added. After stirring overnight the CH2Cl2 layer was washed with 3 x 75 ml of 10% aqueous HCl solution, 3 x 100 ml of saturated sodium bicarbonate under reduced pressure. The crude product was purified on silica gel using ethyl acetate hexanes (1: 5) to give 4.68 g (88%) of a white crystalline solid: m.p. 51-52 ° C, [a] ß ** -14.3ß (c - 2.4, '"CHjOH); IR (Kbr, crn "1) 2948, 2886, 1766, 1745, 1455, 1434; lH NMR (CDCl,) d 7.36-7.19 (m, JL0H), 5.36 (dd, * J» 4.5, 8.7 Hz, 1H); 4.57 (s, 2H), 4.19 (d, J - 16.7 Hz, 18), 4.09 (d, J - 16.7 Hz, 1H), 3.74 (s, 3H), 3.23 (dd, J = 4.5, 14.3 Hz, 1H), 3.11 (dd, J-8.7, 14.3 Hz, 1H); HRMS calculated for CltHj0O5 (M *), 328.1311, Encont.328.1269.
(S) -5-benzyl-3-benzyloxy-4-hydroxy-2 (5H) -furanone. To a three-neck, flame-dried, 100 ml flask, purged with argon, equipped with a low temperature thermometer, 2.38 g (14.73 mmol) of hexamethyldisilizide in 35 ml of anhydrous tetrahydrofuran was added. The solution was cooled to -25 ° C, and 5.9 ml of n-BuLi at 2.5 M (14.73 mmoles) in hexanes were added dropwise while keeping the temperature below -15 ° C. After the addition, the reaction was stirred and maintained between -3 and -5 ° C for 45 minutes and cooled to -78 ° C. (S) -2- (2-benzyloxy) acetyloxy-3-phenylactate methyl (2.3 g, 7.01 mmol) in 10 ml of anhydrous tetrahydrofuran were added dropwise. The reaction was stirred for 75 minutes and quenched at -780C with 25 mL of 10% aqueous HCl solution previously cooled. After warming to room temperature, the aqueous layer was extracted with 3 x 89 ml of ether and the combined organic layers were washed with 3 x 100 ml of brine and dried (Na 2 S 4). The ether was evaporated under reduced pressure to provide 1.98 g of crude white solid which was recrystallized using ether / petroleum ether to give 1.82 g (85%) of a white crystalline solid: m.p. 181-182 ° C; [a] D2s -57.1 »(c-0.9, CHjOH); IR (Kbr, cm'1) 3031 (br), 2719, 1743, 1662, 1454; 2H NMR (CD3C0CDj) d 7.34-7.29 (m 5H), 7.29-7.21 (m, 5H), 4.96 (dd, J-3.7, 6.5 Hz, 1H), 4.87 (d, 18.2 Hz, 1H), 4.79 (d, J • 18.2 Hz, 1H), 3.26 (dd, J-3.7, 14.5 Hz, 1H), 2.88 (dd, J-6.5, 14.5 Hz, 1H); 13C NMR (CD3C0CDj) 169.0, 160.2, 138.2, 136.4, 130.6 (2C), 129.2 (2C), 129.1 (2C), 129.0 (2C), 128.8, 127.6, 121.9, 76.0, 73.6, 38.6; HRMS calculated for CnH ^ O, (M *), 296.1048, Encont.296.1045; analysis calculated for ClMilitO: C, 72.96; H, 5.44; Found C: 72.59; H, 5.53.
(S) -5-benzyl-3,4-dihydroxy-2 (5H) -furanone. 0.2 g of 10% palladium on carbon in 10 ml of methanol was suspended in a 250 ml hydrogenation bottle flushed with argon. To this suspension was added 2.0 g (6.76 mmoles) of (S) -5-benzyl-3-benzyloxy-4-hydroxy-2 (5H) -furanone and 25 ml of methanol. The mixture was stirred at room temperature under hydrogen (2.46 kg / cm) and monitored by CCD (approximately 5-6 hours). After filtration (Celite pad) the filtrate was evaporated under reduced pressure and the residue was purified by recrystallization from acetone / hexane to give 1.25 g (90%) of a white crystalline solid: m.p. 142-144 ° C; [< *] "" -40.2 ° (c = 2.1, CHjOH); IR (Kbr, cnT1) 3334 (br), 1762, 1681, 1455, 1319; aH NMR (CD3COCD3) d 7.28-7.21 (ra, SH), 4.93 (dd, J »3.5, 6.7 HZ, 1H), 3.29 (dd, J * 3.5, 14.5 HZ, 1H), 2.88 (dd, J = 6.5, 14.5 Hz, 1H); HRMS calculated" for CuH10O4 (M *) , 206.0579, Encont «206.0583; analysis calculated for CuH1004: C, 64.08; H, 4.82; Encont.C: 63.99; H, 4.89.
EXAMPLE 8 (R) -3-phenylactate methyl was prepared in a similar yield as for the corresponding S-isomer of (R) -phenylactic acid: m.p. 47 ° C; IR (Kbr, cm "1) 3479 (br), 3029, 2954, 1739, 1496, 1454; - * NMR (CDCl, d 7.34-7.19 (ra, 5H), 4.50-4.43 (m, 1H), 3.78 ( s, 3H), 3.14 (dd, J «4.3, 13.9 Hz, 1H), 2.97 (dd, J = 6.8, 13.9 Hz, 1H), 2.72 (d, J = 6.2 Hz, 1H); HRMS calculated for C10H12O3 ( M *), 180.0786, Enco'nt 180.0795.
(R) -2- (2-benzyloxy) acetyloxy-3-phenylactate methyl was prepared in a similar yield as for the corresponding S-isomer from (R) -3-phenylactate: m.p. 49-50 ° C; (C = 0.4, CH 3 OH); IR (Kbr, was "1) 2948, 2886, 1766, 1745, 1455, 1434, ** H NMR (CDC13) 6 7.35-7.19 (m, 10H), 5.36 (dd, J-4.5, 8.7 Hz, 1H) , 4.57 (s, 2H), 4.19 (d, J * «16.7 Hz, 1H), 4.09 (d, J = 16.7 Hz, 1H), 3.74 (s, 3H)," 3.23 (dd, J »4.5, 14.3 Hz, 1H), 3.11 (dd, J «8.7, 14.3 Hz, 1H); HRMS calculated for CuHjßOj (M *), 328.1311, Encont.328.1303.
(R) -5-benzyl-3-benzyloxy-4-hydroxy-2 (5H) -furanone was prepared in a similar yield as for (S) -5-benzyl-3-benzyloxy-4-hydroxy -2 (5H) -furanone from methyl (R) -2- (2-benzyloxy) acetyloxy-3-phenylactate: mp 182-183 ° C; [ot] 0ss 57. 8β (C »0.4, CH 3 OH); IR (Kbr, cm'1) 3029 (br), 2717, 1743, 1660, 1454, ** H NMR (CDjCOCD3) d 7.34-7.29 (m, 5H), 7.29-7.21 (m, 5H), 4.96 (dd) , J - 3.7, 6.5 Hz, 1H), 4.86 (d, 18.4 Hz, 1H), 4.79 (d, J »18.4 Hz, 1H), 3.26 (dd, J * 2.7, 14.4 Hz, 1H), 2.88 (dd , J • 6.5, 14.4 Hz, 1H); HRMS calculated for Cl $ Hxí04 (M *), 296.1048, Encont, 296.1045.
(R) -5-benzyl-3,4-dihydroxy-2 (5H) -furanone was prepared in a similar yield as for the corresponding S-isomer from (R) -5-benzyl-3-benzyloxy-4- hydroxy-2 (5H) -furanone: mp 142-144 ° C; W 40.8 ° (c = 0.8, CHsOH); IR (Kbr, cm'1) 3336 (br), 1762, 1679, 1455, 1319 .--- H NMR (CD3COCD3) 6 7.28-7.22 (m, 5), 4.93 (dd, J »3.6, 6.7 Hz, 1H), 3.29 (dd, J = 3.6, 14.5 Hz, 1H), 2.88 (dd, J.6.7, 14.5 Hz, 1H); HRMS calculated for C?;. H1004: C, 64.08; H, 4.82. Encont; C, 63.93, H, 4.86. ls EXAMPLE 9 (S) -5- (2-hydroxymethyl) -2,2, dimethyl-1,2-dioxolan-4-one. ? n a round flask with 500 ml flame-dried flask equipped with a partition and under argon was placed 11.0 g (63.2 mmol) of the (S) -malic acid diosolanate (prepared by reaction of commercially available (S) -malonic acid with excess of dimethoxypropane and p-toluenesulfonic acid as a catalyst) dissolved in 200 ml of anhydrous tetrahydrofuran. The reaction was cooled (-20 ° C to -30 ° C) and 70 ml of a 1 M borane-tetrahydrofuran complex was added dropwise over 2 hours. After the addition, the reaction vessel was placed in a refrigerator at 4 ° C for 11 hours, heated to At room temperature, it was stirred at room temperature for 9 hours and chromatographed (silica gel) using acetone as eluent. After evaporation under reduced pressure, the residue was chromatographed as before to generate 9.1 g (90%) of the alcohol as a colorless, unstable liquid which was dried under reduced pressure and used as such in the following reaction: 3480 (br), 2994, 2940, 2888, 1791, 1220; lH RN (CDC13) d 4.58 (dd, J = 5.1, 7.0 HZ, 1H), 3.91-3.79 (ra, 2H), 2.28-2.13 (ra, 1H), 2.13-1.97 (ra, 1H), 1.64 (s) , 3H), 1.57 (s, 3H). p-Toluenesulfonate of (S) -5- (2-hydroxyethyl) -2,2-dimethyl-1,3-dioxolan-4-one. Dry (S) -5- (2-hydroxyethyl) -2,2-dimethyl-1,3-dioxolane-4-one (9.0 g, 59.2 mmol) was dissolved in 100 ml of anhydrous pyridine under argon and cooled to - 4 ° C. To this solution maintained at about 0 ° C was added dropwise 11.3 g (59.22 mmoles) of p-toluenesulfonyl chloride dissolved in 100 ml of pyridine. After the dition, the mixture was placed in the refrigerator (0-4 ° C) overnight. Water (150 ml) was added and the aqueous mixture was extracted with 4 x 200 ml of ether. The ether layers were combined, washed with 3 x 150 ml of water, 3 x 150 ml of saturated copper sulfate solution (until no dark blue remained), 2 x 100 ml of water, 3 x 150 ml of brine, dried (Na2 =? 4), filtered and evaporated under reduced pressure. The crude tosylate was purified by column chromatography using ethyl acetate: hexanes (1: 1) to provide the title compound, 16.7 g (95%) as a white solid: m.p. 48-49 ° C; [α »-3.7» (c = 0.3, CH 3 OH); IR (Kbr, cm'1) 2989, 1785, 1390, 1357, 1278; : H NMR (CDClj) d 7.80 (d, J = 8.3 Hz, 2H), 7.36 (d, J = 8.3 HZ, 2H), 4.43 (dd, J -.4.4, 8.1 Hz, 1H), 4.27-4.14 ( m, 2H), 2.46 (s, 3H), 2.3-2.17 (ra, 1H), 2.09-1.92 (m, 1H), 1.58 (s, 3H), 1.51 (s, 3H); HRMS. calculated for C14HlßOßS (M *), 314.0824, Encont.314.0817.
(S) -5-hexyl-2, 2-dimethyl-l, 3-dioxolan-4-one. To a 300 ml three-neck round bottom flask dried under nitrogen containing a suspension of 4.85 g (25.48 mmol) of cuprous iodide in 200 ml of anhydrous ether and kept at -30 ° C, 31.85 was added dropwise. ml of n-BuLi at 1.6 M (50.96 mmoles) in hexanes. The dark red-brown solution was stirred at -30 ° C to -40 ° C for 2 hours and cooled to -78 ° C. p-Toluenesulfonate of (S) -5- (2-hydroxyethyl) -2, 2-dimethyl-l, 3-dioxolan-4-one (4.0 g, 12.74 mmoles) dissolved in 30 ml of anhydrous ether and 10 ml of tetrahydrofuran was added dropwise while maintaining the temperature below -70 ° C. The reaction mixture was stirred for 18 hours at -78 ° C. After completion (supervision with CCD), the reaction was heated to -10 ° C and quenched by the addition of 125 ml of saturated ammonium chloride solution previously cooled. Ether (100 ml) was added and the mixture was filtered over Celite. The aqueous phase was extracted with 3 x 175 ml of ether. The combined ether extract was washed with 2 x 125 ml of saturated ammonium chloride solution, 1 x 75 ml of water and 2 x 100 ml of brine, dried (Na S04), filtered and concentrated under reduced pressure. The product was purified (silica gel) using ethyl acetate: hexanes (1: 5) as eluent to give 2.44 g (95%) of the title compound as a colorless oil: [a] D2s 0.6 ° (c = 1.9, CHjOH); IR (Kbr, was "1) 2958, 2933, 2861, 1797; * H NMR (CDCl,) d 4.39 (dd, J-4.4, 7.1 Hz, 1H), 1.95-1.81 (m, 1H), 1.81-1.62 (ra, 1H), 1.61 (s, 3H), 1.54 (s) , 3H), 1.49-1.38 (ra, 2H), 1.35-1.29 (m, 6H), 0.89 (t, J - 6.6"C NMR (CDCl,) 6 173.3, 110.2, 74.2, 31.7, 31.6, 28.8, 25.8 , 24.8, 22.5, 13.9; HRMS calculated for CuHaß03 (M *), 200.1412, Encont, 200.1422.
(S) methyl -2-hydroxyoctanoate. In a 250 ml round bottom flask equipped with a reflux condenser were placed 2. 4 g (12 mmol) of (S) -5-hexyl-2,2-dimethyl-l, 3-dioxolan-4-one in 150 ml of methanol containing 2 drops of concentrated sulfuric acid. After heating to reflux for 6 hours, the reaction mixture was cooled, and 0.5 g of sodium bicarbonate was added. The solvent was evaporated under reduced pressure and the residue was dissolved in 200 ml of CH2Cl2. The solution was washed with 2 x 75 ml of water, 2 x 100 ml of saturated sodium bicarbonate solution, and 2 x 75 ml of brine, and dried (Na2 =? 4). The CH2Cl2 solvent was evaporated under reduced pressure to provide 2.07 g of crude product which was purified on silica gel using ethyl acetate hexanes (1: 5) to give 2.0 g (96%) of the title compound as a yellow oil pale: [a] D "2.9 ° (c = 0.8, CKjOH), IR (Net, era-1) 3475 (br), 2925, 2857, 1739; * H NMR (CDClj) d 4.19 (m, 1H), 3.79 (s, 3H), 2.73 (d, J - 4.8 Hz, 1H), 1.89-1.75 (ra, 1H), 1.72-1.56 (m, 1H), 1.55-1.39 (m, 2H), 1.39-1.28 ( m, 6H), 0.88 (t, J - 6.6 Hz, 3H); HRMS calculated for C? A.0, (* », 174.1255, Encont.174.1255.
(S) -2- (2-benzyloxy) acetyloxyoctanoate methyl. In a 250 ml two-necked round bottom flask, flame dried, under argon, 4.28 g (25 or mmoles) of methyl (S) -2-hydroxyoctanoate in 100 ml of anhydrous CH2Cl2 was added. Benzyloxycetyl chloride (6.82 g, 36.9 mmol) was added and the mixture was cooled to 0 ° C in an ice-salt bath. Pyridine (2.92 g, 36.9 mmol) was added dropwise. The mixture was stirred for 30 minutes at 0 ° C, warmed to room temperature, stirred for a further 3 hours, and quenched with 30 ml of ice water. An additional 50 ml of CH CI2 was added. After stirring overnight, the CH2Cl2 layer was separated and extracted with 3 x 50 ml of 10% aqueous HCl solution, 3 x 74 ml of saturated sodium bicarbonate solution, and 2 x 100 ml of salmeter, dried (a2S? 4) and concentrated under reduced pressure. The residue was purified on silica gel with ethyl acetate: hexanes (1: 6) as eluent to give 6.7 g (88%) of the title compound as a pale yellow liquid: [OfJo "-10.9 ° (c = 0.3, CH3OH); IR (Net, cm'1) 2927, 2859, 1735, 1455, 1438, lH NMR (CDC13) d 7.36-7.29 (m, 5H), 5.11 (t, J * 6.5 Hz, 1H), 4.66 ( s, 2H), 4.23 (d,. = 16.7 Hz, 1H), 4.17 (2, J = 16.7 Hz, 1H), 3.75 (s, 3H), 1.87-1.72 (m, 2H), 1.42-1.15 (m , 8H), 0.87 (t, J - 6.6 Hz, 3H); HRMS calculated for C ^ H ^ Os (M *), 322.1780, Encont.322.1760.
(S) -3-benzyloxy-5-hexyl-4-hydroxy-2 (5H) -furanone. In a flame-dried three-neck 100 ml flask equipped with a low temperature thermometer, 2.1 g (13.04 mmoles) of hexamethyldisilazide in 35 ml of anhydrous tetrahydrofuran were added. After cooling to -25 ° C, 5.3 ml of n-Buli at 2.5 M in hexanes (13.04 mmoles) was added dropwise and with stirring while keeping the temperature below -15 ° C. The mixture was stirred and maintained at -3 ° C to -5 ° C for 45 minutes and cooled to -78 ° C. (S) -2- (2-benzyloxy) acetyloxyoctanoate methyl (2.0 g, 6.21 mmol) in 10 ml of anhydrous tetrahydrofuran were added dropwise. The reaction was stirred for 75 minutes and was quenched at -78 ° C with 30 ml of 10% aqueous HCl solution previously cooled. After warming to room temperature the product was extracted with 3 x 80 ml of ether, and the organic layer was washed with 3 x 100 ml of brine, dried (Na 2 SO 4) and evaporated under reduced pressure to give 1.65 g of crude white product which was recrystallized using ether / petroleum ether to give 1.53 g (85%) of white crystals: pg 74-75 ° C; [] a2i -18.9 ° (c «2.0, CHjOH); IR (KBr, crn "1.}. 3035 (br), 2950, 2921, 1735, 1646, 1465; -? NMR (CD3COCD3) d 7.43-7.29 (m, 5H),? .06 (d, 16.1 Hz, 1H), 5.01 (d, J * 16.1 Hz, 1H), 4.69 (dd, J-3.7, 7.0 Hz, 1H ), 1.90-1.82 (ra, 1H), 1.61-1.45 (m, 1H), 1.38-1.15 (m, 8H), 0.87 (t, J »6.8 Hz, 3H); 13C MN (CD3COCD3) d 171.6, 169.4, 161.2, 138.3, 129.2 (2C), 129.1 (2C), 128.9, 120.9, 75.9, 73.4, 32.6, 32.4, 24.4, 23.2, 14.3; HRMS calculated for Ct, H «04 (M *), 290.1518; Encont., 290.1538; analysis calculated for CltHj304 C 70.36, H 7.50; Encont.C 70.32, H 7.64. / -lu (S) -5-hexyl-3,4-dihydroxy-2 (5H) -furanone. In a 100 ml two-necked round bottom flask, 0.7 g was added under argon. (2.41 mmoles) of (S) -3-benzyloxy-4-hydroxy-3 (5H) -furanone, 0.7 g of 10% Pd / C and 4.96 g (660.35 mmoles) of cyclohexane in 50 g. ml of absolute ethanol. The mixture was stirred and heated to reflux, filtered (Celite) and the solvent was removed under reduced pressure. The residue was recrystallized from acetone / hexanes to give 0.362 g (75%) of a white solid: m.p. 100-101 ° C; [α] D «-14.10 (c to 20 0.4, CHjOH), - IR (KBr, cm-1) 3426 (br), 2921, 1766, 1662; lK NMR (CDjC0CD3) or 4.56 (dd, J-3.4, 7.0 Hz, 1H), 1.98-1.84 (ra, 1H), 1.57-1.43 (m, 1H), 1.42-1.22 (m, 8H), 0.90 (t , J - 6.7 Hz, 3H); HRMS calculated for ClcHla4 (Mf), 200.1049; Encont.200.1049; analysis calculated for C ^ A 'C 60.05, H 8.05.
EXAMPLE 10 (R) -5- (2-Hydroxyethyl) -2,2-dimethyl-1,3-dioxolan-4-one was prepared in a similar yield as for the corresponding S-isomer from (S) -malic acid dioxolamate : IR (Net, cm'1) 3453 (br), 2994, 2940, 2888, 1791, 1220; *? fc / MR (CDC13) 4.58 (dd, J = 5.1, 7.0 HZ, 1H), 3.91-3.79 (m, 2H), 2.28-2.13 (m, 1H), 2.12-1.97 (ra, 1H), 1.64 (s) , 3H), 1.57 (s, 3H). p-Toluenesulfonate of (R) -5- (2-hydroxyethyl) -2,2-dimethyl-l, 3-dioxolan-4-one was prepared in a similar yield as for the corresponding S-isomer from (R) -5- (2-hydroxyethyl) -2,2-dimethyl-l, 3-dioxolane-4-one: mp 52-53 ° C; [ar] B "3.8 ° (c * 1.1, CHjOH); IR (KBr, was" 1) 2992, 1785, 1388, 1357, 1278; lK NMR (CDC13) d 7.80 (d, J - 8.3 Hz, 2H), 7.36 (d, J »8.3 Hz, 2H), 4.43 (dd, J * 4.4, 8.1 Hz, 1H), 4.27-4.14 (m, 2H), 2.46 (s, 3H), 2.30-2.18 (m, 1H), 2.09-1.93 (m, 1H), 1.58 (S, 3H), 1.51 (s, 3H); HRMS calculated for CI.HUOÍS (M *), 314.0824; Encont.314.0830.
(R) -5-hexyl-2,2-dimethyl-1,3-dioxolan-4-one was prepared in a similar yield as for the corresponding S-isomer from (R) -5- (2-hydroxyethyl) - 2,2-dimethyl-1,3-dioxolan-4-one: [a) p2i -2.9 ° (c = 1.3, CHjOH); IR (nßat, era'1) 2958, 2931, 2859, 1793; lH NMR (CD3Cls) d 4.39 (dd, J »4.4, 7.1 Hz, 1H), 1.95-1.81 (ra, 1H), 1.81-1.62 (m, 1H), l.βl (s, 3H), 1.54 (s) , 3H), 1.49-1.38 (m, 2H), 1.24-1.28 (ra, 6H), 0.89 (t, J - 6.6 HZ, 3H); HRMS calculated for Ca? H "03 (M *), 200.1412; Encont.200.1431.
(R) -2-hydroxyoctanoate methyl was prepared in a similar yield as for the corresponding S-isomer from (R) -5-hexyl-2,2-dimethyl-l, 3-dioxolan-4-one: Ca] C2S 2.7 ° (c = 0.3, CHjOH); IR (Neto, cpT1) 3496 (br), 2929, 2859, 1749; lH NMR (CDClj) d 4.19 (m, 1H), 3.79 (s, 3H), 2.73 (d, J - 4.8 Hz, 1H), 1.89-1.75 (m, 1H), 1.72-1.57 (m, 1H), 1.55-1.39 (m, 2H), 1.39-1.28 (m, 6H), 0.88 (t, J »6.6 HZ, 3H); HRMS calculated for CjH ^ Os (M *), 174.1255; Encont, 17 .1256. Methyl (R) -2- (2-benzyloxy) acetyloxy was prepared in a similar yield as for the corresponding S-isomer from methyl (R) -2-hydroxyoctanoate: [α] D2S 10.8 ° (c-1.1, CH30H); IR (KBr, crn "1) 2925, 2857, 1733, 1455, 1436; lH NMR (CDCla) d 7.36-7.29 (ra, 5H), 5.10 (t, J * 6.5 Hz, 1H), 4.65 (S, 2H ), 4.23 (d, J - 16.7 Hz, 1H) -, 4.17 (d, J = 16.7 Hz, 1H), 3.74 (s, 3H), 1.87-1.72 (m, 2H), 1.42-1.15 (m, 8H ), 0.87 (t, J - 6.6 HZ, 3H); HRMS calculated for UHJJOS (M *), 322.1780; Encont.322.1782.
(R) -3-benzyloxy-5-hexyl-4-hydroxy-2 (5H) -furanone was prepared in a similar yield as for the corresponding S-isomer for methyl (R) -2- (2-benzyloxy) acetyloxyoctonoate: pf 86-87 ° C; [0f3D2S 18.8 * (c-0.9, CH, OH); IR (KBr, car1) 3035 (br), 2950, 2921, 1735; 1646; 1465; lH MN (CD, COCD,) d 7.43-7.29 (ra, 5H), 5.07 (d, 16.2, 1H), 5.01 (d, J> 16.2, 1H), 4.69 (dd, J - 3.7, 7.0, 1H ), 1.90-1.82 (ra, 1H), 1.61-1.45 (ra, 1H), 1.61-1.45 (m, 1H), 1.3fi-1.15 (m, 8H), 0.87 (t, J = 6.7 HZ, 3H); HRMS calculated for C "HM04 (M *), 290.1518, Encont.290.1505, (R) -5-hexyl-3,4-dihydroxy-2 (5H) -furanone was prepared in a similar yield as for the S-isomer corresponding to starting from (R) -3-benzyloxy-5-hexyl-4-hydroxy-2 (5H) -furanone: mp 98-99 ° C; [α] K 14.2 * (c * 1.9, CH 3 OH); IR (KBr, cpT1) 3423 (br), 2921, 1768, 1660; XH NMR (CD3COCD3) d 4.57 (dd, J = 3.4, 7.0 Hz, 1H), 1.98-1.84 (m, 1H), 1.57-1.43 (m, 1H), 1.42-1.22 (m, 8H), 0.90 (t , J = 6.7 Hz, 3H); HRMS calculated for C10Hls04 (M *), 200.1049; Encont.200.1049.

Claims (34)

NOVELTY OF THE INVENTION CLAIMS
1. - An optically pure compound of the general formulas la or Ib: (la) (Ib) wherein R is an alkenyl group of 2-20 carbon atoms or an alkanyl group of 9-20 carbon atoms, and when an alkenyl group contains one or more degrees of unsaturation, or a physiologically acceptable salt of the same.
2. - A compound in accordance with the claim 1, further characterized in that R is an alkanyl group of 9 to 20 carbon atoms.
3. - A compound in accordance with the claim 2, further characterized in that it is (S) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone.
4. A compound according to claim 2, further characterized in that it is (R) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone.
5. A compound according to claim 1, further characterized in that R is an alkenyl group of 2 to 20 carbon atoms.
6. A compound according to claim 5, further characterized in that it is (S) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
7. A compound according to claim 5, further characterized in that it is (R) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
8. A compound according to claim 5, further characterized in that it is (S) -3,4-dihydroxy-5- [(all-Z) -3, 6, 9, 12-octadecatetraenyl] -2 (5H) -furanone.
9. A compound according to claim 5, further characterized in that it is (R) -3,4-dihydroxy-5- [(all-Z) -3,6,9, 12-octadecatetraenyl] -2 (5H) -furanone.
10. A pharmaceutical composition comprising an effective amount of a compound selected from the group consisting of an optically pure compound of the formulas Ia or Ib: . { Ia) (Ib) wherein R is an alkenyl group of 2-20 carbon atoms or an alkanyl group of 9-20 carbon atoms, and when a --... alkenyl group contains one or more degrees of unsaturation, or a physiologically acceptable salt of the same, in combination with a pharmaceutically acceptable vehicle therefor.
11. A composition according to claim 10, further characterized in that R is an alkanyl group of 2 to 20 carbon atoms.
12. - A composition according to claim 11, further characterized in that it is (S) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone.
13. A composition according to claim 11, further characterized in that it is (R) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone.
14. - A composition according to claim 10, further characterized in that R is an alkenyl group of 2 to 20 carbon atoms.
15. A composition according to claim 14, further characterized in that it is (S) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
16. - A composition according to claim 14, further characterized in that it is (R) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
17. A composition according to claim 14, further characterized in that it is (S) -3,4-dihydroxy-5- [(all-Z) -3, 6, 9, 12-octadecatetraenyl] -2 (5H) - furanone.
18. A composition according to claim 14, further characterized in that it is (R) -3,4-dihydroxy-5- [(all-Z) -3,6,9, 12-octadecatetraenyl] -2 (5H) -furanone.
19. - A process for the preparation of compounds of the formula I wherein R is an alkenyl or alkanyl group of 2-20 carbon atoms, and when an alkenyl group contains one or more degrees of unsaturation, or an aryl group; comprising: (a) coupling of allyloxyacetic acid with the optically pure alkanyl ester of formula II or its corresponding isomer, wherein R 'is as defined above; and ale is a lower alkanyl group of 1 to 6 carbon atoms in the presence of DCC and an acid acceptor such as 4-pyrrolidinopyridine to obtain an allyloxy ester of the formula III or its corresponding isomer, wherein ale and R 'are as defined above; (b) cyclization of the allyl ester of the formula III with LiHMDA to give the allyloxytetronic acid of the formula IV or its corresponding isomer, wherein R 'is as defined above; (c) isomerization of the allyloxy aci-reductone of the formula IV with hydrogen and an iridium catalyst to give the corresponding ether-propenyl of the formula V or its corresponding isomer, wherein R 'is as defined above; and (d) hydrolysis of the enol ether of the formula V, or its corresponding isomer with aqueous acid, to give the desired compound of the formula I.
20. A process according to claim 19, further characterized in that the coupling of the step (a) is carried out using chloride as a solvent.
21. A process according to claim 19, further characterized in that the cyclization of step (b) is carried out using about 2 equivalents of LiHMDA at about -78 ° C.
22. A process according to claim 19, further characterized in that the isomerization of step (c) is carried out using hexafluorophosphate of [bis (methyldiphenylphosphino] (1,5-cyclooctadiene) iridium as the catalyst 23.
A process according to claim 19, further characterized in that the hydrolysis of step (d) is carried out with 50% aqueous acetic acid.
A method for treating or preventing atherosclerosis disorders comprising administering to a mammal in need of such therapy an effective amount of a compound selected from the group consisting of an optically pure compound of the formulas Ia or Ib: (la) (Ib) wherein R is an alkenyl group of 2-20 carbon atoms or an alkanyl group of 9-20 carbon atoms, and when an alkenyl group contains one or more degrees of unsaturation, or a physiologically acceptable salt thereof, in combination with a pharmaceutically acceptable vehicle therefor.
25. A method according to claim 24, further characterized in that R is an alkanyl group of 9 to 20 carbon atoms.
26. A method according to claim 24, further characterized in that (S) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone.
27. A method according to claim 24, further characterized in that (R) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone.
28.- A method in accordance with the claim 24, further characterized in that R is an alkenyl group of 2 to 20 carbon atoms.
29. A method according to claim 24, further characterized in that (S) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
30. A method according to claim 24, further characterized in that (R) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone.
31. A method according to claim 24, further characterized in that it is (S) -3,4-dihydroxy-5- [(all-Z) -3,6,9,1-octadecatetraenyl] -2 (5H) -furanone.
32. - A method according to claim 24, further characterized in that it is (R) -3,4-dihydroxy-5- [(all-Z) -3,6,9,1-octadecatetraenyl] -2 (5H) -furanone .
33. A method for inhibiting inflammatory cytokines comprising administering to a mammal in need of such therapy an effective amount of a compound selected from the group consisting of an optically pure compound of the formulas I or Ib: wherein R is an alkenyl group of 2-20 carbon atoms or an alkanyl group of 9-20 carbon atoms, and when an alkenyl group contains one or more degrees of unsaturation, or a physiologically acceptable salt thereof, in combination with a pharmaceutically acceptable vehicle therefor.
34. A method according to claim 20 33, further characterized in that R is an alkanyl group of 9 a 20 carbon atoms. 35.- A method in accordance with the claim 33, further characterized in that (S) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone. 25 36.- A method in accordance with the claim 33, further characterized in that (R) -3,4-dihydroxy-5-3-octadecanyl-2 (5H) -furanone. 37. A method according to claim 33, further characterized in that R is an alkenyl group of 2 to 20 carbon atoms. 38.- A method in accordance with the claim 33, further characterized in that (S) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone. 39. A method according to claim 33, further characterized in that (R) -3,4-dihydroxy-5- [(Z) -3-octadecenyl] -2 (5H) -furanone. 40. A method according to claim 33, further characterized in that it is (S) -3,4-dihydroxy-5- [(all-Z) -3, 6, 9, 12-octadecatetraenyl] -2 (5H) -furanone. 41. A method according to claim 33, further characterized in that it is (R) -3, -dihydroxy-5- [(all-Z) -3,6, 9, 12-octadecatetraenyl] -2 (5H) - furanone. 42.- A method according to claim 33, further characterized in that the inflammatory cytokine is present in rheumatoid arthritis 43.- A method in accordance with the claim 33, further characterized in that the inflammatory cytokine is present in acute or chronic inflammation. 44. - A method according to claim 33, further characterized in that the inflammatory cytokine is present in multiple sclerosis. 45. A method according to claim 33, further characterized in that the inflammatory cytokine is present in septic shock syndrome. 46. A method according to claim 33, further characterized in that the inflammatory cytokine is present in cachexia.
MXPA/A/1996/005760A 1994-05-19 1995-05-18 Synthesis of 4-alkenil or 4-alcanil-2-hydroxitetronic acids optically pu MXPA96005760A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US08/245,615 US5504107A (en) 1990-01-12 1994-05-19 Optically pure 4-alkenyl- or 4-alkanyl-2-hydroxytetronic acids and pharmaceutical use thereof
US08245615 1994-05-19
PCT/US1995/006220 WO1995032194A1 (en) 1994-05-19 1995-05-18 Synthesis of optically pure 4-alkenyl- or 4-alkanyl-2-hydroxytetronic acids

Publications (2)

Publication Number Publication Date
MX9605760A MX9605760A (en) 1998-05-31
MXPA96005760A true MXPA96005760A (en) 1998-10-23

Family

ID=

Similar Documents

Publication Publication Date Title
RU2540330C2 (en) Bicyclic compound and using it for medical applications
JP2763894B2 (en) Biologically active carboxylic esters
KR101561171B1 (en) Process for the preparation of f-series prostaglandins
JP3992297B2 (en) Vitamin D3 derivative and therapeutic agent for inflammatory respiratory diseases using the same
CN1533372A (en) Process and intermediates to prepare latanoprost
CH638483A5 (en) 2,2-difluoro-6a carba pgi2 compounds.
EP0322004A1 (en) Cyclic anti-inflammatory derivatives of di-tert-butylphenol compounds
WO2006112742A2 (en) PROCESS FOR PREPARATION OF 13,14-DIHYDRO-PGF2α DERIVATIVES
US5504107A (en) Optically pure 4-alkenyl- or 4-alkanyl-2-hydroxytetronic acids and pharmaceutical use thereof
KR20150022831A (en) Dihydroorotic acid dehydrogenase inhibitor
IL87037A (en) 3-Demethyl-4-fluoromevalonic acid derivatives, a process for the preparation thereof, pharmaceutical products based on these compounds, and the use thereof
EP2780336A1 (en) Structural variants of mycolactones for use in modulating inflammation, immunity and pain
JP2002509915A (en) C11 oxymyl and hydroxylamino prostaglandins useful as medicaments
JP2608089B2 (en) 4-hydroxy-2-cyclopentenones and pharmaceutical compositions containing them
MXPA96005760A (en) Synthesis of 4-alkenil or 4-alcanil-2-hydroxitetronic acids optically pu
JPH07508501A (en) Fluorine-containing vitamin D↓3 analogue
US5656662A (en) Synthesis of optically pure 4-alkenyl- or 4-alkanyl-2-hydroxytetronic acids
NZ286456A (en) Cyclohexanol and cyclohexanone derivatives spiro condensed in position-4 with an oxirane ring; n-acyl carbamate esters derived therefrom
EP0746551A1 (en) Optically pure 4-aryl-2-hydroxytetronic acids
SU1072801A3 (en) Process of preparing derivatives of prostacycline or their epimers
FI78289C (en) FOERFARANDE FOER FRAMSTAELLNING AV TERAPEUTISKT ANVAENDBARA BICYKLO / 3.2.0 / HEPTAN-6-ONOXIMETERDERIVAT.
RU2501793C2 (en) Method of producing corey lactone enantiomeric derivatives
US4704462A (en) Substituted 2,3,3a,6-tetrahydro-6-oxobenzofuran derivative useful as PAF antagonist
Ishibashi et al. Synthesis and algicidal activity of (+)-cyanobacterin and its stereoisomer
JPS6296438A (en) 4-hydroxy-2-cyclopentenone and production thereof