CYP-eicosanoid derivatives

Abstract

The present invention relates to compounds according to general formula (I) which are analogues of epoxymetabolites produced by cytochrome P450 (CYP) enzymes from omega-3 (n-3) polyunsaturated fatty acids (PUFAs). The present invention further relates to compositions containing one or more of these compounds and to the use of these compounds or compositions for the treatment or prevention of conditions and diseases associated with inflammation, proliferation, hypertension, coagulation, immune function, pathologic angiogenesis, heart failure and cardiac arrhythmias.

Claims

1. A compound of the general formula (IV): ##STR00073## or a pharmaceutically acceptable salt thereof, wherein P is a group represented by the general formula (II):
—(CH.sub.2).sub.n—B—(CH.sub.2).sub.k—X   (II) wherein X represents: ##STR00074## wherein k is 0 or 1; and B represents a bond, —O—, or —S—; n is 0 or an integer of from 3 to 8; and k is 0 or 1, provided that when n is 0 or 3, then k is 1 and when n is 3, B is —O— or —S—; wherein R.sup.1 represents a hydroxyl group, C.sub.1-C.sub.6alkoxy, —NHCN, —NH(C.sub.1-C.sub.6alkyl), —NH(C.sub.3-C.sub.6cycloalkyl), —NH(aryl), or —O(C.sub.1-C.sub.6alkyldiyl)O(C═O)R.sup.11; R.sup.11 is a C.sub.1-C.sub.6alkyl group which is optionally substituted with one or more fluorine or chlorine atom(s), or with a C.sub.3-C.sub.6cycloalkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); R.sup.2 represents —NHR.sup.3, —NR.sup.20R.sup.21, —OR.sup.22, or —(OCH.sub.2—CH.sub.2).sub.i—R.sup.23; wherein R.sup.3 represents (SO.sub.2R.sup.30), (OR.sup.31), —C.sub.1C.sub.6alkanediyl(SO.sub.2R.sup.32), or —C.sub.1-C.sub.6alkanediyl(CO.sub.2H); R.sup.30 is a C.sub.1-C.sub.6alkyl group which is optionally substituted with —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, C.sub.1-C.sub.6alkylcarbonyloxy-, C.sub.1-C.sub.6alkoxycarbonyloxy-, C.sub.1-C.sub.6alkylcarbonylthio-, C.sub.1-C.sub.6alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-, one, two or three fluorine or chlorine atoms, or a hydroxyl group, or an aryl group which is optionally substituted with one, two or three substituents independently selected from the group consisting of C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6alkylthio, fluorine or chlorine atom, hydroxyl group, amino group, —NH(C.sub.1-C.sub.6alkyl), and —N(C.sub.1-C.sub.6)dialkyl; R.sup.31 is a C.sub.1-C.sub.6alkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); or a C.sub.3-C.sub.6cycloalkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); R.sup.32 is a C.sub.1-C.sub.6alkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s), or a C.sub.3-C.sub.6cycloalkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); R.sup.°and R.sup.21 each independently represents a hydrogen atom, a C.sub.1-C.sub.6alkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s), a C.sub.3-C.sub.6cycloalkyl group which is optionally substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s) or —C.sub.1-C.sub.6alkyldiyl(CO.sub.2H); R.sup.22 is a hydrogen atom, a C.sub.1-C.sub.6alkyl group or a C.sub.3-C.sub.6cycloalkyl group, wherein the C.sub.1-C.sub.6alkyl group or the C.sub.3-C.sub.6cycloalkyl group is optionally substituted with —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, —NH(C.sub.1-C.sub.6)alkyldiyl, —C.sub.1-C.sub.6alkoxy, one, two or three fluorine or chlorine atom(s), hydroxy, or C.sub.1-C.sub.6alkoxy; R.sup.23 is —OH, —O(C.sub.1-C.sub.3)alkyl, or —N(C.sub.1-C.sub.3)dialkyl; i is an integer of from 1 to 10; R.sup.4 represents ##STR00075## h is 0, 1, or 2; R.sup.6 and R.sup.7 each independently represent a hydroxyl group, an —O(C.sub.1-C.sub.6)alkyl group, an —O(C.sub.2-C.sub.6)alkenyl group, a —O(C.sub.1-C.sub.6)alkyldiylO(C═O)(C.sub.1-C.sub.6)alkyl group or a —(C.sub.1-C.sub.6)alkyldiylO(C═O)(C.sub.2-C.sub.6)alkenyl group, wherein the C.sub.1-C.sub.6alkyl group and the C.sub.2-C.sub.6alkenyl group are optionally substituted with NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, C.sub.1-C.sub.5alkylcarbonyloxy-, C.sub.1-C.sub.6alkoxycarbonyloxy-, C.sub.1-C.sub.6alkylcarbonylthio-, C.sub.1-C.sub.6alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-; or with one, two or three fluorine or chlorine atom(s); R.sup.8 and R.sup.8′ each independently represents a hydrogen atom, a C.sub.1-C.sub.6alkyl group, —C(═O)C.sub.1-C.sub.6alkyl, —C(═O)C.sub.3-C.sub.6cycloalkyl, —C(═O)aryl; or —C(═O)heteroaryl, wherein the C.sub.1-C.sub.6alkyl, the C.sub.3-C.sub.6cycloalkyl, the aryl, or the heteroaryl group may be substituted with one, two or three substituents selected from the group consisting of fluorine atom, chlorine atom, hydroxy, —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, -N(C.sub.1-C.sub.6)dialkyl, —NH(C.sub.1-C.sub.6)alkanediyl-C.sub.1-C.sub.6alkoxy, and C.sub.1-C.sub.6alkoxy; X.sup.1 represents an oxygen atom; sulfur atom; or NH; X.sup.2 represents an oxygen atom; sulfur atom; NH; or N(CH.sub.3); wherein R.sup.12 and R.sup.13 of formula (IV) each independently is a hydrogen atom, a fluorine atom, hydroxy, —NH.sub.2, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, —C(═O)-aryl, —C(═O)C.sub.1-C.sub.6alkyl, —SO.sub.2(C.sub.1-C.sub.6alkyl) or —SO.sub.2aryl, wherein any of the foregoing C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, -—(═O)-aryl, or —SO.sub.2aryl are optionally substituted with one, two or three substituents independently selected from the group consisting of: —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, C.sub.1-C.sub.6alkylcarbonyloxy-, C.sub.1-C.sub.6alkoxycarbonyloxy-, C.sub.1-C.sub.6alkylcarbonylthio-, C.sub.1-C.sub.6alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-, fluorine atom, chlorine atom, and hydroxy; or R.sup.12 and R.sup.13 form together a 5-membered or 6-membered ring, which is optionally substituted with one, two or three substituents independently selected from the group consisting of: —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, C.sub.1-C.sub.6alkylcarbonyloxy-, C.sub.1-C.sub.6 6alkoxycarbonyloxy-, C.sub.1-C.sub.6alkylcarbonylthio-, C.sub.1-C.sub.6alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-, fluorine atom, chlorine atom, and hydroxy; I is —(CH.sub.2).sub.3—Y, wherein Y represents a group: ##STR00076## wherein R.sup.40, R.sup.41, R.sup.43, and R.sup.44 each represents a hydrogen atom; R.sup.42 is a methyl group and R.sup.45 is an ethyl group; with the proviso that when: (i) n is 3 and each of R.sup.12 and R.sup.13 is a hydrogen atom; (ii) n is 4 and k is 1 and each of R.sup.12 and R.sup.13 is a hydrogen atom; or (iii) n is 5, 6, 7, or 8, and each of R.sup.12 and R.sup.13 is a hydrogen atom then, P represents one of the following groups: —(CH.sub.2).sub.3—O—(CH.sub.2)—X.sup.81, —(CH.sub.2).sub.3—S—(CH.sub.2)—X.sup.81, —(CH.sub.2).sub.5—O—(CH.sub.2)—X.sup.81, —(CH.sub.2).sub.5—S—(CH.sub.2)—X.sup.81, —(CH.sub.2).sub.5—O—X.sup.82, —(CH.sub.2).sub.5—X.sup.83; —(CH.sub.2).sub.7—O—X.sup.82; or —(CH.sub.2).sub.7—X.sup.83, wherein X.sup.81 represents a group: ##STR00077## X.sup.82 represents a group: ##STR00078## X.sup.83 represents a group: ##STR00079## R.sup.1′ is defined as R.sup.1 above; R.sup.2′ represents —NHR.sup.3′, —OR.sup.22′, —(OCH.sub.2—CH.sub.2).sub.i—R.sup.23, wherein R.sup.3′ represents (SO.sub.2R.sup.30), (OR.sup.31 ), —C.sub.1-C.sub.6alkanediyl(SO.sub.2R.sup.32), or —C.sub.2-C.sub.6alkanediyl(CO.sub.2H), R.sup.22′ is a C.sub.3-C.sub.6cycloalkyl group, which is optionally substituted with —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, —NH(C.sub.1-C.sub.6)alkyldiyl-C.sub.1-C.sub.6alkoxy, one, two or three fluorine or chlorine atom(s), hydroxy, or C.sub.1-C.sub.6alkoxy, R.sup.23 and i are as defined above, provided that when i=3, R.sup.23 is not —OH; R.sup.4′ is defined as R.sup.4 above; and h is defined as above; R.sup.6′ and R.sup.7′ are defined as R.sup.6 and R.sup.7 above; R.sup.8″ and R.sup.′″ are defined as R.sup.8 and R.sup.8′ above; R.sup.9′ is defined as R.sup.9 above; R.sup.9″ represents aryl which is optionally substituted with one, two or three substituents independently selected from the group consisting of C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6alkylthio, fluorine or chlorine atom, hydroxyl group, amino group, —NH(C.sub.1-C.sub.6alkyl), —N(C.sub.1-C.sub.6)dialkyl, and an oxo substituent.

2. The compound according to any one of claim 1, wherein X is ##STR00080##

3. The compound according to claim 1, wherein one of R.sup.12 and R.sup.13 represents a hydrogen atom and the other represents a fluorine atom, hydroxy, —NH.sub.2, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, —C(═O)-aryl, —C(═O)C.sub.1-C.sub.6alkyl, or —SO.sub.2(C.sub.1-C.sub.6alkyl); or —SO.sub.2aryl; wherein any of the foregoing C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, or aryl are optionally substituted with one, two or three substituents independently selected from the group consisting of —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, C.sub.1-C.sub.6alkylcarbonyloxy-, C.sub.1-C.sub.6alkoxycarbonyloxy-, C.sub.1-C.sub.6alkylcarbonylthio-, C.sub.1-C.sub.6alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-, fluorine or chlorine atom, and hydroxy; or R.sup.12 and R.sup.13 are taken together to form a 5-membered or 6-membered ring, which ring is optionally substituted with one, two or three substituents independently selected from the group consisting of —NH.sub.2, —NH(C.sub.1-C.sub.6)alkyl, —N(C.sub.1-C.sub.6)dialkyl, C.sub.1-C.sub.6alkylcarbonyloxy-, C.sub.1-C.sub.6alkoxycarbonyloxy-, C.sub.1-C.sub.6alkylcarbonylthio-, C.sub.1-C.sub.6alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-, fluorine or chlorine atom, and hydroxy.

4. The compound according to claim 1, wherein P is —(CH.sub.2).sub.3—O—(CH.sub.2)—X.sup.81; —(CH.sub.2).sub.5—O—(CH.sub.2)—X.sup.81; —(CH.sub.2).sub.3—S—(CH.sub.2)—X.sup.81; —(CH.sub.2).sub.5—S—(CH.sub.2)—X.sup.81; —(CH.sub.2).sub.5—O—X.sup.82; —(CH.sub.2).sub.7—O—X.sup.82; —(CH.sub.2).sub.5—X.sup.83 or —(CH.sub.2).sub.7—X.sup.83.

5. (Withdrawn- Currently Amended) The compound according to claim 4, wherein P represents —(CH.sub.2).sub.5—X.sup.83 or —(CH.sub.2).sub.7—X.sup.83.

6. The compound according to claim 1, wherein X.sup.83 represents a group selected from the groups consisting of: ##STR00081##

7. The compound according to claim 1, wherein n is 3 and R.sup.1′ is a hydroxyl group; and R.sup.2′ represents —NHR.sup.3′ wherein R.sup.3′ is (SO.sub.2R.sup.30), R.sup.30 is —C.sub.1-C.sub.6alkyl or phenyl; R.sup.6′ and R.sup.7′ each independently represents a hydroxyl group; an —O(C.sub.1-C.sub.6)alkyl group; or an —O(CH.sub.2)O(C═O)(C.sub.1-C.sub.6)alkyl group; R.sup.8″ is hydrogen atom; and R.sup.8′″ is —C(═O)C.sub.1-C.sub.6alkyl.

8. A compound selected from the group consisting of: ##STR00082## ##STR00083##

9. A pharmaceutical composition that comprises at least one compound according to claim 1, and, optionally, a carrier substance and/or an adjuvant.

10. The compound according to claim 1, wherein said compound is part of a medicament.

11. A method comprising: administering to a patient that suffers from a cardiovascular disease or has suffered in the past from a cardiovascular disease an effective amount of at least one of the compounds of claim 1.

12. The method of claim 11, wherein the cardiovascular disease is ventricular arrhythmia or atrial fibrillation.

13. The method of claim 12, wherein at least 0.5 mg of the at least one compound is administered to the patient in one day.

14. A method for providing omega-3 (n-3) polyunsaturated fatty acids (PUFA) derivatives to a persons at risk to develop a condition or disease associated with inflammation, proliferation, hypertension, coagulation, immune function, pathologic angiogenesis, or cardiac disease comprising: providing orally or parenterally to the person at risk at least one of the compounds of claim 1 in an amount considered effective to reduce the risk of developing said condition or disease.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1: Examples of n-3 PUFA analogues according to the present invention and their effect on an established in vitro cardiac arrhythmia model using spontaneously beating neonatal rat cardiomyocytes (NRCMs) compared to eicosapentaenoic acid (EPA) and 17,18-epoxyeicosatetraenoic acid (17,18-EEQ). As demonstrated, spontaneous beating of the cells under basal conditions was reduced by the application of the example analogs.

(2) The following examples serve to more fully describe the manner of using the above-described invention, as well as to set forth the best modes contemplated for carrying out various aspects of the invention. It is understood that these examples in no way serve to limit the true scope of this invention, but rather are presented for illustrative purposes.

EXAMPLES

(3) Specific examples for the preparation of compounds of formula (I) are provided in the following examples. Unless otherwise specified all starting materials and reagents are of standard commercial grade, and are used without further purification, or are readily prepared from such materials by routine methods. Those skilled in the art of organic synthesis will recognize that starting materials and reaction conditions may be varied including additional steps employed to produce compounds encompassed by the present invention. Preferred methods include but are not limited to those methods described below. Each of the references cited in connection with the described route of synthesis is hereby incorporated herein by reference.

Example 1

(4) Preparation of Intermediate 10

Step A. Synthesis of tert-butyldiphenyl((15-((tetrahydro-2H-pyran-2-yl)oxy)pentadec-5-yn-1-yl)oxy)silane (3)

(5) ##STR00039##

(6) n-BuLi (2.5 M in hexanes, 1 eqiv) was added dropwise to a −78° C. solution of 1 (1 eqiv) in anhydrous THF and freshly distilled HMPA (3:1). The reaction was stirred at −78° C. for 30 min and at 0° C. for 2 h. The reaction was re-cooled to −78° C. and 2 (1.2 eqiv) in dry THF was added slowly. After 40 min at −78° C. and at rt overnight (14 h), the reaction was quenched with sat. NH.sub.4Cl solution, water was added and the reaction was extracted twice with EtOAc. The combined organic extracts were washed twice with water, dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum. Purification via SiO.sub.2 column chromatography using 2% EtOAc/hexanes afforded 3 (89%) as a colorless oil.

(7) .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.70-7.63 (m, 4H), 7.47-7.30 (m, 6H), 4.61-4.54 (m, 1H), 3.92-3.82 (m, 1H), 3.78-3.70 (m, 1H), 3.67 (t, J=6.2 Hz, 1H), 3.56-3.45 (m, 1H), 3.44-3.33 (m, 1H), 2.22-2.07 (m, 2H), 1.90-1.39 (m, 10H), 1.39-1.23 (m, 9H), 1.04 (s, 9H), 0.92-0.81 (m, 2H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 135.80 (4), 134.26 (2), 129.76 (2), 127.84 (4), 99.08, 80.67, 80.22, 67.90, 63.73, 62.57, 31.99, 31.05, 30.01, 29.75, 29.73, 29.41, 29.39, 29.13, 27.12 (3), 26.48, 25.82, 25.77, 19.97, 19.47, 19.02, 18.81.

Step B. Synthesis of 15-((tetrahydro-2H-pyran-2-yl)oxy)pentadec-5-yn-1-ol (4)

(8) ##STR00040##

(9) To a solution of 3 in dry THF was added tetra-n-butylammonium fluoride (TBAF, 1.0 M soln in THF, 1.3 equiv). After 39 h, the THF was evaporated, the residue was suspended in water, and extracted twice with Et.sub.2O. The organic extracts were dried over MgSO.sub.4, filtered, and concentrated in vacuo. The residue was purified using a Teledyne Isco Combiflash® RF chromatographic system to give alcohol 4 (61%) as a colorless oil.

(10) .sup.1H NMR (500 MHz, CDCl.sub.3) δ 4.57 (dd, J=4.5, 2.8 Hz, 1H), 3.91-3.83 (m, 1H), 3.72 (dt, J=9.5, 6.9 Hz, 1H), 3.67 (t, J=6.4 Hz, 2H), 3.54-3.44 (m, 1H), 3.37 (dt, J=9.5, 6.7 Hz, 1H), 2.19 (tt, J=6.9, 2.4 Hz, 2H), 2.13 (tt, J=7.1, 2.4 Hz, 2H), 1.89-1.77 (m, 1H), 1.77-1.40 (m, 13H), 1.40-1.24 (m, 10H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 98.98, 80.75, 79.96, 67.86, 62.50 (2), 32.03, 30.93, 29.90, 29.61, 29.26 (2), 29.01, 26.39, 25.67, 25.60 (2), 19.83, 18.90, 18.73.

Step C. Synthesis of 15-(tetrahydro-2H-pyran-2-yloxy)pentadec-5(Z)-en-1-ol (5)

(11) ##STR00041##

(12) To a suspension of Ni(OAc).sub.2 (0.6 equiv) in absolute ethanol in a two necked flask under H.sub.2 (1 atm) was added NaBH.sub.4 (0.8 equiv) in one portion. After 25 min, distilled ethylenediamine (EDA, 3 equiv) was added neat followed by a solution of 4 in absolute EtOH. After 2 h, the reaction mixture was filtered through a pad of silica gel. The pad was washed with EtOAc. The combined organic filtrates were concentrated to give olefin 5 (98%), obtained as a colorless oil. TLC: 20% EtOAc/hexanes, R.sub.f˜0.35. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 5.41-5.31 (m, 2H), 4.59-4.56 (m, 1H), 3.89-3.85 (m, 1H), 3.73 (dt, J=9.7, 6.9 Hz, 1H), 3.65 (t, J=6.5 Hz, 2H), 3.53-3.47 (m, 1H), 3.38 (dt, J=9.7, 6.9 Hz, 1H), 2.06 (dt, J=7.0, 6.5 Hz, 2H), 2.01 (dt, J=7.0, 6.5 Hz, 2H), 1.87-1.79 (m, 1H), 1.75-1.68 (m, 1H), 1.64-1.48 (m, 9H), 1.46-1.38 (m, 2H), 1.38-1.24 (m, 11H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 130.5, 129.6, 99.0, 67.9, 62.8 (2), 62.5, 32.6, 30.9, 29.9, 29.8(2), 29.7, 29.5, 27.4, 27.2, 26.4, 26.1, 25.6, 19.8.

Step D. Synthesis of 2-(15-azidopentadec-10(Z)-enyloxy)tetrahydro-2H-pyran (6)

(13) ##STR00042##

(14) To a −25° C. solution of triphenylphosphine (TPP, 1.2 equiv) in dry THF was added dropwise diisopropyl azodicarboxylate (DIAD, 1.2 equiv). Ten minutes later, a solution of alcohol 5 in dry THF was added dropwise at the same temperature to form a yellow suspension. After 30 min, the reaction mixture was warmed to 0° C. and then diphenylphosphoryl azide (DPPA, 1.2 equiv) was added dropwise. The reaction mixture was warmed to rt and stirred. After 16 h, the reaction mixture was quenched with H.sub.2O and extracted with Et.sub.2O three times. The combined ethereal extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The crude product was purified via SiO.sub.2 column chromatography using 2% EtOAc/hexanes as eluant to give 6 (>97%) as a light yellow oil. An analytical sample was further purified using preparative TLC to give 6 as a colorless oil. TLC: 20% EtOAc/hexanes, R.sub.f˜0.8. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.28 (m, 2H), 4.59-4.56 (m, 1H), 3.91-3.84 (m, 1H), 3.77-3.69 (m, 1H), 3.54-3.46 (m, 1H), 3.41-3.35 (m, 1H), 3.27 (t, J=6.8 Hz, 2H), 2.10-1.95 (m, 4H), 1.88-1.78 (m, 1H), 1.76-1.68 (m, 1H), 1.65-1.48 (m, 6H), 1.46-1.38 (m, 2H), 1.38-1.24 (m, 14H); .sup.13C NMR (100 MHz, CDCl.sub.3) δ 130.9, 129.0, 99.0, 67.8, 62.4, 51.5, 31.0, 29.9, 29.8, 29.7, 29.6 (2), 29.5, 28.6, 27.4, 26.9, 26.8, 26.4, 25.7, 19.9.

Step E. Synthesis of 15-(tetrahydro-2H-pyran-2-yloxy)pentadec-5(Z)-en-1-amine (7)

(15) ##STR00043##

(16) To a rt solution of the above crude azide 6 in THF was added triphenylphosphine (TPP, 1.3 equiv) in one portion. After 2 h, H.sub.2O was added and the reaction was stirred at rt. After 12 h, the reaction mixture was diluted with EtOAc followed by brine and the biphasic mixture was extracted with EtOAc three times. The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The crude product was triturated in Et.sub.2O and filtered through a fritted funnel. The filtrate was concentrated in vacuo and the crude 7 was utilized in the next step without further purification. TLC: 5% MeOH/CH.sub.2Cl.sub.2, R.sub.f˜0.1.

Step F. Synthesis of N.SUP.1.-methyl-N.SUP.2.-(15-(tetrahydro-2H-pyran-2-yloxy)pentadec-5(Z)-enyl)oxalamide (9)

(17) ##STR00044##

(18) Following literature precedent,.sup.1 the above crude amine 7 and 8 (1.2 equiv) in anhydrous absolute ethanol were heated at 85° C. in a sealed tube. After 15 h, the reaction mixture was concentrated in vacuo and the crude product was purified via SiO.sub.2 column chromatography using 25% EtOAc/hexanes to give 9 (70%) as a white solid, mp 69.9-70.2° C. TLC: 50% EtOAc/hexanes, R.sub.f˜0.65. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.56-7.46 (br s, 2H), 5.42-5.28 (m, 2H), 4.59-4.56 (m, 1H), 3.91-3.84 (m, 1H), 3.72 (dt, J=9.6, 6.9 Hz, 1H), 3.54-3.46 (m, 1H), 3.37 (dt, J=9.6, 6.7 Hz, 1H), 3.30 (app q, J=6.9 Hz, 2H), 2.91 (d, J=5.5 Hz, 3H), 2.04 (dt, J=7.5, 7.0 Hz, 2H), 1.98 (app q, J=7.0, 2H), 1.88-1.78 (m, 1H), 1.76-1.68 (m, 1H), 1.64-1.47 (m, 7H), 1.44-1.22 (m, 15H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 160.8, 159.9, 130.9, 129.1, 99.1, 67.9, 62.6, 39.8, 31.0, 29.9(2), 29.8, 29.7(2), 29.5, 29.0, 27.5, 27.1, 26.9, 26.5, 26.4, 25.7, 19.9.

Step G. Synthesis of N.SUP.1.-(15-hydroxypentadec-5(Z)-enyl)-N.SUP.2.-methyloxalamide (10)

(19) ##STR00045##

(20) To a solution of 9 in methanol was added p-toluenesulfonic acid (PTSA, 0.07 eqiv). After 2 h, the solvent was evaporated in vacuo and the residue was re-dissolved in EtOAc. Passage of the crude product through a short silical gel pad using EtOAc as eluant gave 10 (>95%) as a white solid, mp 115.4-115.7° C. TLC: 50% EtOAc/hexanes, R.sub.f˜0.25. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.45 (br s, 2H), 5.42-5.28 (m, 2H), 3.71-3.55 (m, 2H), 3.31 (app q, J=6.8 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.12-1.91 (m, 4H), 1.61-1.52 (m, 6H), 1.44-1.23 (m, 12H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 160.8, 159.9, 130.9, 129.1, 63.3, 39.8, 33.0, 29.8, 29.7, 29.6(2), 29.4, 29.0, 27.4, 27.1, 26.9, 26.4, 25.9.

Example 2

(21) Preparation of Example Compound C41

Step A. Synthesis of N.SUP.1.-(15-bromopentadec-5(Z)-enyl)-N.SUP.2.-methyloxalamide (11)

(22) ##STR00046##

(23) To a solution of TPP (745 mg, 1.2 equiv) in CH.sub.2Cl.sub.2 (80 mL) under an argon atmosphere was added a solution of common intermediate 10 (740 mg, 2.37 mmol, 1 equiv) in CH.sub.2Cl.sub.2 (40 mL) followed by carbon tetrabromide (CBr.sub.4, 1.2 equiv, 942 mg) in one portion. After 24 h, the reaction mixture was concentrated under vacuum and the residue was purified via silica gel column chromatography using 20-25% EtOAc/hexanes to give 11 (597 mg, 67%) as a white solid, mp 77.5-77.6° C. TLC: 50% EtOAc/hexanes, R.sub.f˜0.7. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.45 (br s, 2H), 5.42-5.27 (m, 2H), 3.41 (t, J=6.8 Hz, 2H), 3.31 (app q, J=6.8 Hz, 2H), 2.91 (d, J=5.6 Hz, 3H), 2.09-1.96 (m, 4H), 1.85 (app quintet, J=7.2 Hz, 2H), 1.62-1.52 (m, 2H), 1.47-1.37 (m, 4H), 1.37-1.23 (m, 10H); .sup.13C NMR (100 MHz, CDCl.sub.3) δ 160.8, 159.9, 130.9, 129.1, 39.8, 34.3, 33.0, 29.9, 29.6(2), 29.5, 29.0, 28.9, 28.4, 27.4, 27.1, 26.9, 26.4.

Step B. Synthesis of dimethyl 15-(2-(methylamino)-2-oxoacetamido)pentadec-10(Z)-enylphosphonate (12)

(24) ##STR00047##

(25) A solution of 11 (375 mg, 1.1 mmol) and trimethyl phosphite [P(OMe).sub.3] (16 mL) in dry THF (16 mL) was heated in a sealed tube at 120° C. After 3 d, the THF was evaporated in vacuo and the P(OMe).sub.3 was distilled off under reduced pressure. The crude 12 (240 mg, 54%) was subjected to the next reaction without further purification. An analytical sample was purified by preparative TLC. TLC: 50% EtOAc/hexanes, R.sub.f˜0.2. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.48 (br s, 2H), 5.39-5.26 (m, 2H), 3.72 (d, J.sub.P,H=10.5 Hz, 6H), 3.30 (app q, J=7.0 Hz, 2H), 2.90 (d, J=5.0 Hz, 3H), 2.04 (app q, J=7.5 Hz, 2H), 1.98 (app q, J=7.5 Hz, 2H), 1.81-1.67 (m, 2H), 1.63-1.51 (m, 4H), 1.43-1.21 (m, 14H); .sup.31P NMR (202 MHz, CD.sub.3OD; rel 85% H.sub.3PO.sub.4) δ 36.48 (s).

Step C. Synthesis of disodium 15-(2-(methylamino)-2-oxoacetamido)pentadec-10(Z)-enylphosphonate (C41)

(26) ##STR00048##

(27) Following literature precedent,.sup.2 TMSBr (10 equiv, 0.5 mL) was added dropwise to a 0° C. solution of 12 (150 mg, 0.371 mmol) in anhydrous CH.sub.2Cl.sub.2 (10 mL). After 75 min, the reaction was quenched with methanol (5 mL), concentrated in vacuo, and the residue was triturated with CH.sub.2Cl.sub.2 (2×10 mL). The residue, mp 130.6-130.7° C., was dissolved in aq. Na.sub.2CO.sub.3 solution (0.01 M, pH 10). Bio-Rad™ SM-2 Bio-Beads (20-50 mesh, 5 g) were added to the solution. After gently stirring for 30 min, the beads were collected on a fritted funnel and washed with water (20 mL). Methanol was then used to strip C41 from the Bio-Beads. Evaporation of the methanol afforded C41 (48 mg, 30%) as a white powder, mp 240° C. (dec). Free acid of C41: .sup.1H NMR (500 MHz, CD.sub.3OD) δ 8.58 (br s, 3H), 5.44-5.24 (m, 2H), 3.26 (app q, J=6.5 Hz, 2H), 2.82 (s, 1H), 2.81 (s, 2H), 2.12-1.98 (m, 4H), 1.88-1.72 (m, 2H), 1.71-1.50 (m, 4H), 1.47-1.24 (m, 14H); .sup.31P NMR (202 MHz, CD.sub.3OD; rel 85% H.sub.3PO.sub.4) δ 31.37 (s). C41: .sup.1H NMR (500 MHz, CD.sub.3OD) δ 5.43-5.28 (m, 2H), 3.26 (t, J=7.0 Hz, 2H), 2.82 (s, 3H), 2.14-1.95 (m, 4H), 1.88-1.71 (m, 2H), 1.69-1.42 (m, 4H), 1.42-1.24 (m, 14H).

Example 2A

(28) Preparation of Example Compounds C52 and C53

(29) ##STR00049##

(30) To a stirring, rt solution of dimethyl phosphonate 12 (1.0 mmol, 0.418 g) in dry CH.sub.3CN (10 mL) and CH.sub.2Cl.sub.2 (2 mL) under an argon atmosphere was added pivaloyloxymethyl iodide (POM-I; purchased from Enamine LLC, Princeton Corporate Plaza, 7 Deer Park Drive, Ste. M-3, Monmouth Jct., N.J. 08852 USA) (5.0 mmol, 0.76 mL). After 2 days, most the starting phosphonate was consumed (TLC analysis: 5% MeOH/CH.sub.2Cl.sub.2). The volatiles were evaporated in vacuo and the crude product was purified by SiO.sub.2 flash column chromatography using a gradient of 1-2% MeOH in CH.sub.2Cl.sub.2to give pure mono-POM ester C53 (20 mg, 4%) as an oil and di-POM ester C52 with some impurities. A second purification using preparative TLC (5% MeOH/CH.sub.2Cl.sub.2) furnished pure di-POM ester C52 (21 mg, 3%) as an oil. C52. TLC: R.sub.f˜0.5, 5% MeOH/CH.sub.2Cl.sub.2; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.46 (br s, 2H), 5.64 (d, J.sub.H-P=13.1 Hz, 4H), 5.40-5.23 (m, 2H), 3.28 (app q, J=7.0 Hz, 2H), 2.88 (d, J=5.2 Hz, 3H), 2.02 (app q, J=6.9 Hz, 2H), 1.97 (app q, J=6.9 Hz, 2H), 1.84-1.74 (m, 2H), 1.64-1.49 (m, 4H), 1.40-1.17 (m, 14H), 1.21 (s, 18H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 176.86, 160.53, 159.64, 130.57, 128.83, 81.24 (d, .sup.2J.sub.C-O-P=6.2 Hz), 39.52, 38.70, 30.40 (d, .sup.2J.sub.C-C-P=18.0 Hz), 29.64, 29.40, 29.27, 29.21, 29.02 (.sup.4J.sub.C-P=1.4 Hz), 28.78, 27.20, 26.84, 26.82, 26.65, 26.23 (d, .sup.1J.sub.C-P=84.0 Hz), 26.12, 21.91 (d, .sup.3J.sub.C-P=5.4 Hz). C53. TLC: R.sub.f˜0.4, 5% MeOH/CH.sub.2Cl.sub.2; .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.48 (br s, 2H), 5.68 (d, J.sub.H-P=13.2 Hz, 2H), 5.43-5.27 (m, 2H), 3.74 (d, J.sub.H-P=11.2 Hz, 3H), 3.32 (app q, J=6.9 Hz, 2H), 2.92 (d, J=5.2 Hz, 3H), 2.06 (app q, J=6.9 Hz, 2H), 2.00 (app q, J=6.9 Hz, 2H), 1.83-1.76 (m, 2H), 1.64-1.51 (m, 4H), 1.44-1.20 (m, 14H), 1.21 (s, 9H); .sup.13C NMR (101 MHz, CDCl.sub.3) δ 177.00, 160.55, 159.67, 130.59, 128.84, 81.66 (d, .sup.2J.sub.C-O-P=6.0 Hz), 51.81 (d, .sup.2J.sub.C-O-P=7.2 Hz), 39.53, 38.72, 30.49 (d, .sup.2J.sub.C-C-P=17.5 Hz), 29.66, 29.42, 29.28, 29.22, 29.04 (d, .sup.4J.sub.C-P=1.3 Hz), 28.79, 27.20, 26.90, 26.83, 26.66, 25.83 (d, .sup.1J.sub.C-P=139.4 Hz), 26.14, 22.10 (d, .sup.3J.sub.C-P=5.4 Hz).

Example 3

(31) Preparation of Example Compound C38

Step A. Synthesis of N.SUP.1.-(15-cyanopentadec-5(Z)-enyl)-N.SUP.2.-methyloxalamide (13)

(32) ##STR00050##

(33) To a solution of bromide 11 (550 mg, 1.47 mmol) in DMSO (30 mL) was added KCN (500 mg, 5 equiv) in one portion. After 24 h at rt, the reaction mixture was diluted with water (60 mL) and extracted with EtOAc (20 mL×3). The combined organic extracts were washed with water (25 mL×2) and then with brine (30 mL). The extracts were dried over anhydrous Na.sub.2SO.sub.4, filtered, and then concentrated in vacuo. The residue was purified via SiO.sub.2 column chromatography using 20-25% EtOAc/hexanes to give 13 (490 mg, 99%) as a white powder, mp 88.8-88.9° C.

(34) TLC: 50% EtOAc/hexanes, R.sub.f˜0.55. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.43 (br s, 2H), 5.44-5.23 (m, 2H), 3.31 (app q, J=6.8 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.34 (t, J=7.2 Hz, 2H), 2.09-1.96 (m, 4H), 1.71-1.61 (m, 2H), 1.61-1.49 (m, 4H), 1.49-1.36 (m, 4H), 1.36-1.22 (m, 8H); .sup.13C NMR (100 MHz, CDCl.sub.3) δ 160.8, 159.9, 130.8, 129.1, 120.1, 39.8, 29.8, 29.6, 29.5, 29.4, 29.0, 28.9, 28.8, 27.4, 27.1, 26.9, 26.4, 25.6, 17.3.

Step B. Synthesis of N.SUP.1.-16-amino-16-(hydroxyimino)hexadec-5(Z)-enyl-N.SUP.2.-methyloxalamide (14)

(35) ##STR00051##

(36) Following literature precedent,.sup.3 a solution of nitrile 13 (100 mg, 0.311 mmol), NH.sub.2OH.HCl (108 mg, 5 equiv), and Na.sub.2CO.sub.3 (181 mg, 5.5 equiv) in anhydrous methanol (2 mL) was heated in a sealed tube at 84° C. After 2 d, the reaction mixture was cooled to rt, filtered, and concentrated in vacuo. The residue was triturated with EtOAc (60 mL×3) and then water (70 mL). The white solid residue (76 mg, 69%) was used in the next reaction without further purification. An analytical sample was purified by preparative TLC (5% MeOH/CH.sub.2Cl.sub.2, R.sub.f˜0.35), mp 118.1-118.5° C.

(37) .sup.1H NMR (400 MHz, CD.sub.3OD) δ 5.35 (td, J=5.9, 4.6 Hz, 2H), 3.25 (t, J=7.1 Hz, 2H), 2.82 (s, 3H), 2.12-1.98 (m, 6H), 1.90 (s, 1H), 1.56 (app quintet, J=7.3 Hz, 4H), 1.44-1.26 (m, 16H); .sup.13C NMR (100 MHz, CD.sub.3OD) δ 161.2, 160.4, 156.5, 130.1, 129.1, 39.19, 30.61, 29.63, 29.43, 29.38, 29.25, 29.12, 28.97, 28.67, 27.14, 26.94, 26.85, 26.58, 25.08.

Step C. Synthesis of N.SUP.1.-methyl-N.SUP.2.-(15-(2-oxido-3H-1,2,3,5-oxathiadiazol-4-yl)pentadec-5(Z)-en-1-yl)oxalamide (C38)

(38) ##STR00052##

(39) Following literature precedent,.sup.3 pyridine (43.6 μL, 2.6 equiv) followed by a solution of SOCl.sub.2 (20 μL, 1.3 equiv) in CH.sub.2Cl.sub.2 (1 mL) were added to a 0° C. solution of 14 (74 mg, 0.21 mmol) in THF (4 mL). After 1 hr 40 min, all volatiles were removed in vacuo and the residue was diluted with water (10 mL), and extracted with EtOAc (15 mL×5). The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by preparative TLC (10% MeOH/CH.sub.2Cl.sub.2) to give C38 (55 mg, 63%) as a white solid, mp 92.7-92.9° C.

(40) TLC: 5% MeOH/CH.sub.2Cl.sub.2, R.sub.f˜0.6. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 8.38 (s, 1H), 7.50 (s, 2H), 5.41-5.28 (m, 2H), 3.41-3.21 (m, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.62 (t, J=7.7 Hz, 2H), 2.06 (app q, J=7.0 Hz, 4H), 2.01 (app q, J=7.0 Hz, 4H), 1.69 (app quintet, J=7.7 Hz, 2H), 1.63-1.55 (m, 3H), 1.46-1.13 (m, 9H); .sup.13C NMR (100 MHz, CDCl.sub.3) δ 160.72, 159.91, 152.89, 130.89, 129.19, 39.95, 29.43, 29.32, 29.21, 29.12, 29.04, 28.97, 28.88, 27.08 (2), 26.88, 26.58, 26.53, 23.95.

Example 4

(41) Preparation of Example Compound C42

Step A. Synthesis of W-(15-iodopentadec-5(Z)-en-1-yl)-N.SUP.2.-methyloxalamide (15)

(42) ##STR00053##

(43) To a solution of 10 (1.80 g, 5.76 mmol), TPP (1.66 g, 1.1 equiv) and imidazole (784 mg, 2 equiv) in dry THF (180 mL) at 0° C. was added I.sub.2(1.75 g, 1.2 equiv). The reaction was allowed to warm to rt and stirred. After 15 h, the reaction was quenched with sat. NaHSO.sub.3 solution and washed twice with water. The aqueous phase was re-extracted with EtOAc (20 mL×2). The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The residue was purified via SiO.sub.2 column chromatography using 20-25% EtOAc/hexanes to give 15 (1.77 g, 70%) as a white solid, mp 81.7° C. TLC: 50% EtOAc/hexanes, R.sub.f˜0.65. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ 7.43 (br s, 2H), 5.42-5.27 (m, 2H), 3.31 (app q, J=6.9 Hz, 2H), 3.19 (t, J=7.0 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.05 (dt, J=7.5, 7.0 Hz, 2H), 2.00 (app q, J=7.0 Hz, 2H), 1.82 (app quintet, J=7.2 Hz, 2H), 1.63-1.48 (m, 2H), 1.44-1.22 (m, 14H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 160.81, 159.94, 130.87, 129.12, 39.82, 33.80, 30.75, 29.93, 29.67, 29.63, 29.50, 29.06, 28.78, 27.48, 27.11, 26.94, 26.44, 7.68.

Step B. Synthesis of Sodium 15-(2-(methylamino)-2-oxoacetamido)pentadec-10(Z)-en-1-sulfonate (C42)

(44) ##STR00054##

(45) Iodide 15 (200 mg, 0.46 mmol), Na.sub.2SO.sub.3 (231 mg, 4 equiv), ethanol (95%, 3 mL), cyclohexene (0.93 mL, 20 equiv) and water (1.5 mL) were heated at 85° C. in a sealed tube. After 4 d, the reaction mixture was cooled to rt, concentrated under vacuum, dissolved in H.sub.2O, and isolated by adsorption onto Bio-Rad SM-2 Bio-Beads as described for C41 to give C42 (51 mg, 27%) as an off-white solid, mp 202-210° C. (dec). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.86-8.55 (m, 2H), 5.45-5.18 (m, 2H), 3.18-2.99 (m, 2H), 2.65 (d, J=5.9 Hz, 3H), 2.34 (t, J=8.0 Hz, 2H), 2.05-1.87 (m, 4H), 1.60-1.35 (m, 4H), 1.35-1.10 (m, 14H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) δ 161.26, 160.51, 130.34, 129.85, 52.20 (2), 39.23, 29.81, 29.70, 29.58, 29.59, 29.32, 29.10, 28.98, 27.28, 27.07, 26.94, 25.78.

Example 5

(46) Preparation of Example Compound C43

Synthesis of N.SUP.1.-(15-((2-acetamidobenzo[d]thiazol-7-yl)oxy)pentadec-5(Z)-en-1-yl)-N.SUP.2.-methyloxalamide (C43)

(47) ##STR00055##

(48) A sealed tube containing iodide 15 (200 mg, 0.458 mmol), N-(4-hydroxybenzo[d]thiazol-2-yl)acetamide.sup.4 (122 mg, 1 equiv), and K.sub.2CO.sub.3 (95 mg, 1.5 equiv) was heated at 85° C. After 6 h, the reaction was cooled to rt, diluted with EtOAc (15 mL) and water (15 mL), and extracted with EtOAc (15 mL×3). The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The residue was purified on a Teledyne Isco Combiflash® RF chromatographic system (1.2 g SiO.sub.2 column eluted with 50-60% EtOAc/hexane) to give C43 (68 mg, 29%) as a brown solid. The brown solid was dissolved in EtOH (1 mL) and sonicated at rt for 5 mins. Analog C43 precipitated as a white solid, upon standing and drying under high vacuum. TLC: 50% EtOAc/hexanes, R.sub.f˜0.2. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ 11.33 (br s, 1H), 7.85 (br s, 1H), 7.59 (br s, 1H), 7.40 (d, J=8.0 Hz,1H), 7.24 (app t, J=8.0 Hz,1H), 6.89 (d, J=8.0 Hz, 1H), 5.40-5.26 (m, 2H), 4.13 (t, J=6.5 Hz, 2H), 3.32 (app q, J=7.0 Hz, 2H), 2.89 (d, J=5.0 Hz, 3H), 2.24 (s, 3H), 2.04 (app q, J=7.0 Hz, 2H), 1.97 (app q, J=7.0 Hz, 2H), 1.88-1.79 (m, 2H), 1.62-1.51 (m, 2H), 1.49-1.34 (m, 4H), 1.34-1.17 (m, 10H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 169.20, 160.87, 160.15, 158.24, 151.58, 138.33, 133.73, 130.96, 129.07, 125.00, 113.63, 108.24, 69.00, 39.85, 29.83, 29.80, 29.70, 29.65, 29.50, 29.48, 29.02, 27.44, 27.07, 26.90, 26.53, 26.35, 23.59.

Example 6

(49) Preparation of Example Compound C48

(50) Step A.

(51) ##STR00056##

(52) A solution of 16 (12 mmol) and 17 (10 mmol) in absolute EtOH (100 mL) was heated under reflux. After 12 h, the reaction mixture was cooled to rt and concentrated in vacuo to approximately 20% of the original volume when 18 began to precipitate as an off-white solid. The solid was collected by filtration and used in the next step without further purification. TLC: EtOAc/hexanes (2:1), R.sub.f˜0.5; .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.26 (s, 1H), 2.90 (d, J=5.2 Hz, 3H), 4.06-4.13 (m, 2H), 7.44 (br s, 1H), 7.68 (br s, 1H).

(53) Step B.

(54) ##STR00057##

(55) To a solution of Pd(PPh.sub.3).sub.4 (3 mol %, 350 mg) and Cul (5 mol %, 100 mg) in Et.sub.3N (40 mL) under an argon atmosphere was added a solution of 1,2-diiodobenzene (10 mmol, 3.3 g) and 5-hexyne-1-ol (10 mmol, 980 mg) in Et.sub.3N (10 mL). The reaction was heated to 60° C. for 12 h, then cooled to rt and filtered through a pad of Celite®. The filtrate was concentrated in vacuo and the residue was purified using a Teledyne Isco Combiflash® RF chromatographic system [40 g SiO.sub.2 column eluted with EtOAc/hexanes (1:2)] to give 19 (1.5 g, 50%) as a pale yellow oil. TLC: EtOAc/hexanes (1:2), R.sub.f˜0.2. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 1.73-1.81 (m, 2H), 1.82-1.88 (m, 2H), 2.55 (t, J=7.0 Hz, 2H), 3.76 (t, J=7.0 Hz, 2H), 6.98 (dd, J=7.0, 8.0 Hz, 1H), 7.29 (dd, J=7.0, 8.0 Hz, 1H), 7.42 (d, J=7.5 Hz, 1H), 7.84 (d, J=8.0 Hz, 1H).

(56) Step C.

(57) ##STR00058##

(58) Et.sub.3N (16.6 mmol, 2.3 mL) and alcohol 19(1.66 mmol, 500 mg) were added sequentially to a solution of 18 (1.66 mmol, 232 mg), Pd(PPh.sub.3).sub.2Cl.sub.2 (3 mol %, 35 mg) and CuI (5 mol %, 16 mg) in dry CH.sub.3CN (15 mL) under an argon atmosphere. After heating at 50° C. for 12 h, the reaction mixture was cooled to it and filtered through a Celite® pad. The filtrate was concentrated in vacuo and the residue was purified using a Teledyne Isco Combiflashe RF chromatographic system [40 g SiO.sub.2 column eluted with EtOAc/hexanes (2:1)] to afford 20 (362 mg, 70%) as a pale yellow oil. TLC: EtOAc/hexanes (2:1), R.sub.f˜0.15. .sup.1H NMR (500 MHz, CDCl.sub.3) d 1.68-1.78 (m, 2H) 1.80-1.88 (m, 2H), 2.53 (t, J=7.0 Hz, 2H), 2.92 (d, J=5.0 Hz, 3H), 3.76 (t, J=6.5 Hz, 2H), 4.40 (d, J=6.0 Hz, 2H), 7.18-7.28 (m, 2H), 7.36-7.44 (m, 2H), 7.62 (br s, 1H), 8.24 (br s, 1H).

(59) Step D.

(60) ##STR00059##

(61) A mixture of diyne 20 (100 mg) and PtO.sub.2 (10 mg) in dry MeOH (10 mL) was shaken in a Parr hydrogenation apparatus under a H.sub.2 atmosphere (50 psi). After 12 h, the reaction mixture was filtered through a Celite® pad and the filtrate was concentrated in vacuo to give crude 21 as a white solid that was used in the next step without further purification.

(62) Step E.

(63) ##STR00060##

(64) PPh.sub.3 (0.38 mmol, 100 mg) was added in one portion to a 0° C. solution of 21 (0.32 mmol, 100 mg) and CBr.sub.4 (0.48 mmol, 160 mg) in CH.sub.2Cl.sub.2 (5 mL). After stirring at rt for 12 h, the solvent was evaporated in vacuo and the residue was purified using a Teledyne Isco Combiflash® RF chromatographic system [24 g SiO.sub.2 column eluted with EtOAc/hexanes (2:1)] to give bromide 22 (98 mg, 80%) as a white solid. TLC: EtOAc/hexanes (4:1), R.sub.f˜0.7. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.35-1.50 (m, 4H), 1.54-1.62 (m, 2H), 1.80-1.90 (m, 4H), 2.58 (dd, J=8.0, 8.0 Hz, 2H), 2.65 (dd, J=8.0, 8.0 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 3.38 (dd, J=7.2, 7.2 Hz, 2H), 3.40 (J=7.2, 7.2 Hz, 2H), 7.10-7.16 (m, 4H), 7.42 (br s, 1H), 7.47 (br s, 1H); .sup.13C NMR (100 MHz, CDCl.sub.3) δ 26.4, 28.3, 29.0, 30.0, 30.8, 31.2, 32.7, 32.9, 34.2, 39.7, 126.3, 126.5, 129.3, 129.5, 138.8, 140.4, 160.0, 160.7.

(65) Step F.

(66) ##STR00061##

(67) A mixture of bromide 22 (0.54 mmol, 200 mg) and P(OMe).sub.3 (16.2 mmol, 1.9 mL) was heated under reflux in a sealed tube for 48 h, then cooled to rt and the excess P(OMe).sub.3 was removed under vacuum. The residue was purified by PTLC using EtOAc/hexanes/MeOH (2:1:0.3) to give dimethyl phosphonate 23 (195 mg, 88%) as a white solid. TLC: EtOAc/hexanes/MeOH (2:1:0.3), R.sub.f˜0.3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.32-1.42 (m, 4H), 1.50-1.76 (m, 6H), 1.78-1.88 (m, 2H), 2.53-2.58 (m, 2H), 2.60-2.65 (m, 2H), 2.89 (d, J=5.2 Hz, 3H), 3.36 (dd, J=6.8, 6.8 Hz, 2H), 3.71 (d, J.sub.P-H=10.4 Hz, 6H), 7.08-7.12 (m, 4H), 7.56 (br s, 2H).

(68) Step G.

(69) ##STR00062##

(70) TMSBr (2 mmol, 260 uL) was added dropwise to a solution of crude diester 23 (0.19 mmol, 80 mg) in dry CH.sub.2Cl.sub.2 (3 mL) under an argon atmosphere. After 3 h, the reaction was quenched with MeOH (2 mL). After stirring for another 1 h, all volatiles were removed in vacuo and aq. Na.sub.2CO.sub.3 solution (0.5 M) was added to reach pH˜10. Bio-Rad™ SM-2 Bio-Beads (20-50 mesh, 5 g) were added to the solution. After gently stirring for 30 min, the beads were collected on a fritted funnel and washed with water (20 mL). Methanol and EtOAc were then used to strip compound from the Bio-Beads. Evaporation of the organic washes gave C48 (37 mg, 45%) as an off-white solid, mp>300° C. (dec). .sup.1H NMR (400 MHz, CD.sub.3OD) δ 1.34-1.66 (m, 10H), 1.74-1.84 (m, 2H), 2.58 (dd, J=8.0, 8.0 Hz, 2H), 2.63 (dd, J=8.0, 8.0 Hz, 2H), 2.81 (s, 3H), 3.26-3.34 (m, 2H), 7.02-7.12 (m, 4H); .sup.13C NMR (100 MHz, CD.sub.3OD) δ 24.4 (d, J.sub.C-P=4.0 Hz), 24.9, 29.3, 29.5, 30.5, 30.6, 31.3, 31.6 (d, J.sub.C-P=17.4 Hz), 32.4, 39.1, 125.4, 125.6, 128.7, 129.0, 138.9, 140.3, 160.3, 161.0; .sup.31P NMR (162 MHz, CD.sub.3OD; ref 85% H.sub.3PO.sub.4) δ 24.4.

Example 7

(71) Preparation of Example Compound C49

(72) Step A.

(73) ##STR00063##

(74) Crude 24 was prepared closely following the procedure used above to generate homolog 18 and was used without further purification.

(75) Step B.

(76) ##STR00064##

(77) NaH (60 wt % in mineral oil, 714 mmol, 2.85 g) was added in one portion to ethylenediamine (35 mL) at 0° C. under an argon atmosphere. The reaction was stirred at rt for 1 h then at 60° C. for 1 h. After cooling to rt, alcohol 25 (17.85 mmol, 2.84 mL) was added dropwise. Upon complete addition, the reaction mixture was reheated to 60° C. After 1 h, the reaction mixture was cooled to 0° C. and quenched with 1 N HCl. The organic layer was extracted with ether (3×100 mL). The combined ethereal extracts were concentrated in vacuo and the residue was purified using a Teledyne Isco Combiflash® RF chromatographic system [40 g SiO.sub.2 column eluted with EtOAc/hexanes (1:5)] to give 26 (1.4 g, 56%) as a light yellow oil.

(78) Step C.

(79) ##STR00065##

(80) The cross-coupling to generate 27 was conducted as described for the synthesis of 19.

(81) Step D.

(82) ##STR00066##

(83) Following the procedure used to prepare 20, iodide 27 and acetylene 24 were transformed into 28, obtained as a pale yellow solid. TLC: EtOAc/hexanes (2:1), R.sub.f˜0.1. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.32-1.42 (m, 4H), 1.46-1.66 (m, 6H), 2.46 (t, J=6.8 Hz, 2H), 2.72 (t, J=6.8 Hz, 2H), 2.90 (d, J=5.2 Hz, 3H), 3.58 (dd, J=5.2, 13.2 Hz, 2H), 3.64 (t, J=6.4 Hz, 2H), 7.15-7.22 (m, 2H), 7.34-7.40 (m, 2H), 7.46 (br s, 1H), 7.90 (br s , 1H).

(84) Step E.

(85) ##STR00067##

(86) Following the procedure used to prepare 21, diyne 28 was transformed into 29, obtained as a white solid. TLC: EtOAc/hexanes (2:1), R.sub.f˜0.15. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.25-1.40 (m, 10H), 1.50-1.65 (m, 8H), 2.54-2.59 (m, 2H), 2.60-2.65 (m, 2H), 2.90 (d, J=5.2 Hz, 3H), 3.30-3.38 (m, 2H), 3.60-3.68 (m, 2H), 7.08-7.14 (m, 4H), 7.45 (br s, 2H).

(87) Step F.

(88) ##STR00068##

(89) Following the procedure used to prepare 22, alcohol 29 was transformed into 30, obtained as a white solid. TLC: EtOAc/hexanes (2:1), R.sub.f˜0.75. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.22-1.48 (m, 12H), 1.50-1.70 (m, 6H), 1.80-1.90 (m, 2H), 2.52-2.68 (m, 4H), 2.89 (d, J=4.8 Hz, 3H), 3.26-3.46 (m, 4H), 7.08-7.16 (m, 4H), 7.53 (br s, 2H); .sup.13C NMR (100 MHz, CDCl.sub.3) δ 26.4, 28.4, 28.6, 29.0, 29.5, 29.6, 29.7, 29.9, 31.5, 32.3, 32.9, 33.0, 34.3, 39.8, 126.0, 126.2, 129.3, 129.4, 139.7, 140.7, 159.9, 160.8.

(90) Step G.

(91) ##STR00069##

(92) Following the procedure used to prepare 23, bromide 30 was transformed into 31, obtained as a white solid. TLC: EtOAc/hexanes/MeOH (1:1:0.2), R.sub.f˜0.2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.20-1.80 (m, 20H), 2.56-2.70 (m, 4H), 2.93 (s, 3H), 3.30-3.42 (m, 2H), 3.76 (d, J.sub.P-H=10.0 Hz, 6H), 7.08-7.18 (m, 4H), 7.52 (br s, 2H).

(93) Step H.

(94) ##STR00070##

(95) Following the procedure used to prepare C48, dimethyl phosphonate 31 was converted into disodium salt C49, obtained as a white solid, mp>300° C. (dec). .sup.1H NMR (400 MHz, CD.sub.3OD) δ 1.25-1.40 (m, 10H), 1.45-1.65 (m, 10H), 2.56-2.66 (m, 4H), 2.80 (s, 3H), 3.25-3.30 (m, 2H), 7.02-7.12 (m, 4H); .sup.13C NMR (100 MHz, CD.sub.3OD) δ 24.1 (d, J.sub.C-P=4.2 Hz), 24.8, 28.4, 28.7, 28.8, 29.2, 29.3, 29.4, 30.1, 31.3, 31.4 (d, J.sub.C-P=17.4 Hz), 31.8, 32.2, 38.9, 125.4, 125.5, 128.8, 128.9, 139.5, 140.1, 160.2, 161.0; .sup.31P NMR (162 MHz, CD.sub.3OD) d 24.6.

Example 8

(96) Preparation of Example Compound C50

Synthesis of N.SUP.1.-(16-phenylsulfonamido-16-oxohexadec-5(Z)-en-1-yl)-N.SUP.2.-methyloxalamide C50

(97) ##STR00071##

(98) 16-(2-(methylamino)-2-oxoacetamido)hexadec-11(Z)-enoic acid 32 was prepared following literature precedent..sup.5 (Z)-16-(2-(methylamino)-2-oxoacetamido)hexadec-11-enoic acid 32 (30 mg, 0.091 mmol) and benzenesulfonamide 33 (13 mg, 0.091 mmol) were taken in a dried round bottom flask in 5 mL anhydrous DMF under an argon atmosphere. Dimethylaminopyridine (DMAP, 13 mg, 0.12, 1.2 equiv) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (16 mg,0.12 mmol; EDCI. HCl) were added as a solid. After stirring for 12 h at room temperature, the reaction mixture was diluted with water (20 mL) and the combined aqueous layers were extracted with EtOAc (3×20 mL), organic layers were washed with water (2×10 mL) and brine (10 mL). The combined organic extracts were dried over Na.sub.2SO.sub.4, concentrated under reduced pressure, and the residue was purified by PTLC using 100% EtOAc as eluent to give amide (35 mg, 84%) as a white solid. TLC: 100% EtOAc, Rf: 0.30. .sup.1H NMR (CDCl.sub.3, 500 MHz): δ 8.05 (d, J=7.5 Hz, 2H), 7.85 (bs, 1H, NH), 7.65 (bs, 1H, NH), 7.60 (t, J=7.5 Hz, 1H), 7.55 (t, J=7.5 Hz, 2H), 5.26-5.42 (m, 2H), 3.28_3.35 (m, 2H), 2.90 (s, 3H), 2.36 (t, 2H, J=7.3 Hz), 1.97-2.08 (m, 4H), 1.51-1.64 (m, 4H), 1.22-1.42 (m, 14H). mp: 72° C.-73° C.

Example 9

(99) Preparation of Example Compound C44

Synthesis of (Z)—N.SUP.1.-(15-((2-hydroxyphenyl)thio)pentadec-5-en-1-yl)-N.SUP.2.-methyloxalamide C44

(100) ##STR00072##

(101) To a suspension of (Z)—N.sup.1-(15-iodopentadec-5-en-1-yl)-N.sup.2-methyloxalamide (15) (400 mg, 0.92 mmol) and KHCO.sub.3 (1.2 equiv, 1.10 mmol, 111 mg) in anhydrous DMF (3.5 mL) was added 2-mercaptophenol (1 equiv, 116 mg) dropwise. The reaction was stirred overnight at room temperature. Note: The reaction went from a white suspension to a clear solution by the next day. After the reaction was judged complete by TLC analysis, the reaction was quenched with water, extracted with ethyl acetate (3×30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The crude product was purified using a Teledyne Isco Combiflash® RF chromatographic system (12 g SiO.sub.2 column eluted with 15-20% EtOAc/hexane) to give the title phenol (317 mg, 79%) as a pale yellow solid. TLC: 50% EtOAc/hexanes, R.sub.f≈0.65. .sup.1H NMR (CDCl.sub.3, 500 MHz) δ 7.46 (dd, J=7.5, 1.5 Hz, 2H), 7.45-7.39 (brs, 1H), 7.29-7.22 (m, 1H), 6.98 (dd, J=8.3, 1.3 Hz, 1H), 6.87 (td, J=7.5, 1.3 Hz, 1H), 6.78 (s, 1H), 5.42-5.26 (m, 2H), 3.31 (q, J=6.9 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.72-2.65 (m, 2H), 2.10-1.91 (m, 5H), 1.62-1.49 (m, 5H), 1.44-1.23 (m, 12H); .sup.13C NMR (125 MHz, CDCl.sub.3) δ 160.69, 159.81, 156.99, 135.93, 130.95, 130.72, 128.96, 120.74, 119.37, 114.78, 39.68, 36.85, 29.77, 29.73, 29.52, 29.51, 29.34, 29.19, 28.91, 28.67, 27.32, 26.96, 26.79, 26.28. mp: 62.4-62.7° C.

REFERENCES

(102) 1. Meddad-Belhabich, N.; Aoun, D.; Djimdé, A.; Redeuilh, C.; Dive, G.; Massicot, F.; Chau, F.; Heymans, F.; Lamouri, A. Design of new potent and selective secretory phospholipase A.sub.2 inhibitors. 6-Synthesis, structure-activity relationships and molecular modeling of 1-substituted-4-[4,5-dihydro-1,2,4-(4H)-oxadiazol-5-one-3-yl(methyl)]-functionalized aryl piperazin/one/dione derivatives. Bioorg. Med. Chem. 2010, 18, 3588-3600. 2. Borbas, K. E.; Ling Kee, H.; Holten, D.; Lindsey, S. J. A compact water-soluble porphyrin bearing an iodoacetamido bioconjugatable site. Org. Biomol. Chem., 2008, 6, 187-194. 3. Ellingboe, J. W.; Lombardo, L. J.; Alessi, T. R.; Nguyen, T. T.; Guzzo, F.; Guinosso, C. J.; Bullington, J.; Browne, E. N. C.; Bagli, J. F. Antihyperglycemic activity of novel naphthalenylmethyl-3H-1,2,3,5-oxathiadiazole 2-oxides. J. Med. Chem. 1993, 36, 2485-2493. 4. Thiel, O. R.; Bernard, C.; King, T.; Dilmeghani-Seran, M.; Bostick, T.; Larsen, R. D.; Margaret M. Faul, M. M. J. Org. Chem. 2008, 73, 3508-3515. 5. Falck, J. R.; Wallukat, G.; Puli, N.; Gall, M.; Arnold, C.; Konkel, A.; Rothe, M.; Fischer, R.; Müller, D. N.; Schunck, W. H. 17(R),18(S)-epoxyeicosatetraenoic acid, a potent eicosapentaenoic acid (EPA) derived regulator of cardiomyocyte contraction: structure-activity relationships and stable analogues. J. Med. Chem. 2011, 54, 4109-4118. 6. Y. Hamada et al./Bioorg. Med. Chem. Lett. 18: 1649-1653, 2008.

Example 10

(103) Determination of Biological Activities of Selected Example Compounds of the Present Invention

(104) Materials and Methods:

(105) The structures of all compounds tested are given in FIG. 1. The compounds include analogues synthesized as described in Examples 2-9. EPA and 17,18-EEQ (purchased from Cayman Chemical) were used as controls. Before use the compounds to be tested were prepared as 1000-fold stock solutions in ethanol.

(106) In order to measure the biological activities of the novel compounds an established cell model was used (Kang, J. X. and A. Leaf, Effects of long-chain polyunsaturated fatty acids on the contraction of neonatal rat cardiac myocytes. Proc Natl Acad Sci USA, 1994. 91(21): p. 9886-90.). The spontaneously beating neonatal rat cardiomyocytes (NRCMs) are a model system to investigate anti-arrhythmic effects of test-compounds. Irregular and asynchronous beating of the cells in response to arrhythmic substances serve as an in vitro equivalent to cardiac fibrillation in vivo, which can be reversed by synthetic 17,18-EEQ analogs/test compounds.

(107) Isolation and cultivation of NRCMs were performed as described previously (Wallukat, G; Wollenberger, A. Biomed Biochim Acta. 1987;78:634-639; Wallukat G, Homuth V, Fischer T, Lindschau C, Horstkamp B, Jupner A, Baur E, Nissen E, Vetter K, Neichel D, Dudenhausen J W, Haller H, Luft F C. J Clin Invest. 1999; 103: 945-952). Briefly, neonatal Wistar rats (1-2 days old) were killed in conformity to the recommendations of the Community of Health Service of the City of Berlin and the cardiomyocytes were dissociated from the minced ventricles with a 0.2% solution of crude trypsin. The isolated cells were then cultured as monolayers on the bottom (12.5 cm.sup.2) of Falcon flasks in 2.5 ml of Halle SM 20-I medium equilibrated with humidified air. The medium contained 10% heat-inactivated FCS and 2 μmol/l fluoro-deoxyuridine (Serva, Heidelberg, Germany), the latter to prevent proliferation of non-muscle cells. The NRCMs (2.4×10.sup.6 cells/flask) were cultured at 37° C. in an incubator. After 5 to 7 days, the NRCMs formed spontaneously beating cell clusters. The cells in each cluster showed synchronized contraction with a beating rate of 120 to 140 beats per minute. On the day of the experiment, the culture medium was replaced by 2.0 ml fresh serum-containing medium. Two hours later, the beating rates were monitored at 37° C. using an inverted microscope equipped with a heating stage. To determine the basal rate, 6 to 8 individual clusters were selected and the number of contractions was counted for 15 sec. After that, the compound to be tested was added to the culture and the beating rate of the same clusters was monitored 5 min later again. Based on the difference between the basal and compound-induced beating rate of the individual clusters, the chronotropic effects (Δ beats/min) were calculated and are given as mean±SE values. N refers to the number of clusters monitored which originated, in general, from at least three independent NRCM cultures.

(108) Results:

(109) The results of these experiments are presented in FIG. 1. All compounds tested were added to the NRCMs at a final concentration of 30 nM and the measurement was performed after 5 min of incubation; except EPA that was used at a final concentration of 3.3 μM and the effect was monitored after 30 min of incubation. Under the same conditions, the vehicle control (0.1% ethanol) showed no effect on the spontaneous beating rate.

(110) As summarized in FIG. 1, synthetic analogues tested showed a negative chronotropic effect similar to that of EPA and 17,18-EEQ. Therefore, the carboxy group can be replaced with different carboxylic acid bioisoters (C38, C41, C42, C43, C44, C49, C50, C52) without a change in the negative chronotropic effect of these synthetic analogs. Since C44 showed the lowest activity (−7.5±4.5; n=12) it seems to harbor the least effective carboxylic acid bioisostere.

(111) C38, C41, C42, C43, C44, C50 and C52 provide examples for compounds according to the general formula (IV) in claim-1. The location of the double bond in these compounds is in agreement with previous structure-activity relationship studies showing that the 11,12-double bond is essential for the biological activity of 17,18-EEQ and its agonists (Falck J R, Wallukat G, Puli N, Goli M, Arnold C, Konkel A, Rothe M, Fischer R, Müller D N, Schunck W H. 17(R),18(S)-epoxyeicosatetraenoic acid, a potent eicosapentaenoic acid (EPA) derived regulator of cardiomyocyte contraction: structure-activity relationships and stable analogues. J Med Chem. 2011 Jun. 23; 54(12):4109-18). C48 and C49 contain aromatic ring structure in those part of the molecule that otherwise harbors the 11,12-double bond. The negative chronotropic effects of C48 and C49 demonstrate that also compounds according to the general formula (III) in claim-1 are bioactive.