METABOLICALLY ROBUST ANALOGS OF CYP-EICOSANOIDS FOR THE TREATMENT OF CARDIAC DISEASE

Abstract

The present invention relates to compounds according to general formula (I) which are metabolically robust analogues of bioactive lipid mediators derived from omega-3 polyunsaturated fatty acids (n-3 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 cardiovascular diseases.

Claims

1. A compound of the general formula (I):
P-E-I(I) or a pharmaceutically acceptable salt thereof, wherein P is a group represented by the general formula (II):
(CH.sub.2).sub.nO(CH.sub.2).sub.kX(II) wherein n is an integer of from 3 to 8; and k is 0, 1 or 2; most preferably k is 1; X represents CH.sub.2OH, CH.sub.2OAc, CH(O) or a group selected from the group consisting of: ##STR00078## ##STR00079## preferably X is ##STR00080## wherein R and R each independently represents a hydrogen atom; or a C.sub.1-C.sub.6alkyl group which may be substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); 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(CO)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 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; (OCH.sub.2CH.sub.2).sub.iR.sup.23; C.sub.3-C.sub.10-heterocyclyl optionally substituted with one, two or three substituents independently selected from the group consisting of hydroxyl group, C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6alkyl, and oxo; -(Xaa).sub.o; a mono-, or disaccharide, or a derivative thereof, which is joined to C(O) by an ester bond via the 1-O, 3-O, or 6-O-position of the saccharide; or is selected from the group consisting of: ##STR00081## wherein R.sup.3 represents (SO.sub.2R.sup.30); (OR.sup.31); C.sub.1-C.sub.6alkanediyl(SO.sub.2R.sup.32); C.sub.1-C.sub.6alkanediyl(CO.sub.2H), an aryl group, a heteroaryl group, a cycloalkyl group or a heterocycloalkyl group, wherein the aryl group 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 C(O)OR.sup.51; wherein the heteroaryl group, 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 C(O)OR.sup.51; where the cycloalkyl group 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 C(O)OR.sup.51; and wherein the heterocycloalkyl group 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 C(O)OR.sup.51; R.sup.30 is a C.sub.1-C.sub.6alkyl, or an aryl group, wherein the C.sub.1-C.sub.6alkyl group 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; and wherein the aryl group 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.20 and R.sup.21 each independently represents a hydrogen atom; a C.sub.1-C.sub.6alkyl group which may be substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); a C.sub.3-C.sub.6cycloalkyl group which may be substituted with one or more fluorine or chlorine atom(s) or hydroxyl group(s); C.sub.1-C.sub.6alkyldiyl(CO.sub.2H) or together form a C.sub.3-C.sub.10-heterocycloalkyl which may be substituted with one or more C.sub.1-C.sub.6alkyl group(s), C.sub.1-C.sub.6alkoxy group(s), fluorine or chlorine atom(s) or hydroxyl group(s); 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), hydroxyl, or C.sub.1-C.sub.6alkoxy, an aralkyl group, a heteroalkyl group or a heteroalkylcycloalkyl group; 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.24, R.sup.25, and R.sup.26 each independently represents a hydrogen atom; C(O)C.sub.11-C.sub.21alkyl; or C(O)C.sub.11-C.sub.21alkenyl; R.sup.27 represents OH; O(CH.sub.2).sub.2NH.sub.2, OCH.sub.2[CH(NH.sub.2)(CO.sub.2H)], O(CH.sub.2).sub.2N(CH.sub.3).sub.3; or ##STR00082## Xaa represents Gly, a conventional D,L-, D- or L-amino acid, a non-conventional D,L-, D- or L-amino acid, or a 2- to 10-mer peptide; and is joined to C(O) by an amide bond; o is an integer of from 1 to 10; R.sup.4 is selected from the group consisting of: ##STR00083## h is 0, 1, or 2; R.sup.5 represents a hydrogen atom; a fluorine or chlorine atom; CF.sub.3; C(O)OR.sup.51; NHC(O)R.sup.52; C(O)NR.sup.53R.sup.54; or S(O.sub.2)OH; R.sup.51 represents 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), hydroxyl, or C.sub.1-C.sub.6alkoxy; R.sup.52, R.sup.53 and R.sup.54 each independently represents a C.sub.1-C.sub.6alkyl group which is optionally substituted with one or more fluorine or chlorine atom(s); a C.sub.3-C.sub.6cycloalkyl group which is optionally substituted with one or more fluorine or chlorine atom(s); 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.6haloalkyl, 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; R.sup.6 and R.sup.7 each independently represents 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(CO)(C.sub.1-C.sub.6)alkyl group, or a O(C.sub.1-C.sub.6)alkyldiylO(CO)(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 may be 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.6 alkylaminocarbonyl-, di(C.sub.1-C.sub.6)alkylaminocarbonyl-, or one, two or three fluorine or chlorine atom(s); or R.sup.6 represents a hydroxyl group and R.sup.7 represents a group: ##STR00084## R.sup.9 represents C.sub.1-C.sub.6alkyl, or aryl; wherein the C.sub.1-C.sub.6alkyl 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, C.sub.1-C.sub.6alkoxy, aryl, aryloxy, C(O)-aryl, C(O)C.sub.1-C.sub.6alkoxy; and wherein the aryl group 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; g is 1 or 2; 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); X.sup.3 represents an oxygen atom; sulfur atom; nitrogen atom; carbon atom; or COH; and the dashed line represents a carbon-carbon bond or a carbon-carbon double bond; E is a group represented by the general formula (III) or (IV): ##STR00085## wherein R12 and R13 are preferably in cis configuration, and wherein ring A in formula (III) represents a 5-membered or 6-membered carbocyclic or heterocyclic ring containing at least one double bond, including an aromatic carbocyclic or heterocyclic ring, which can be substituted with one to three or one to four 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; and L and T each independently represents a ring atom, wherein L and T are adjacent to another; R.sup.12 and R.sup.13 each independently represents a hydrogen atom, a fluorine atom, hydroxyl, 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, 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 hydroxyl; 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, 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 hydroxyl; I is (CH.sub.2).sub.mY, wherein m is an integer of from 3 to 6, provided that m is an integer of from 3 to 5 when E is a group according to general formula (III); Y represents a group selected from the group consisting of: ##STR00086## wherein R.sup.40, R.sup.41, R.sup.43, R.sup.44, R.sup.46, R.sup.48 and R.sup.49 each independently represents a hydrogen atom, C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.6cycloalkyl, C.sub.1-C.sub.6alkoxy, C(O)aryl, or C(O)C.sub.1-C.sub.6alkyl, wherein any of the foregoing C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.6cycloalkyl, 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.40 and R.sup.41, or R.sup.43 and R.sup.44, are taken together to form a 5-membered or 6-membered ring, which ring may be 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 hydroxyl; R.sup.42, R.sup.45, R.sup.47 and R.sup.50 each independently represents a C.sub.1-C.sub.3alkyl, wherein the C.sub.1-C.sub.3alkyl may be substituted with one, two or three substituents independently selected from the group consisting of NH.sub.2, NH(C.sub.1-C.sub.3)alkyl, N(C.sub.1-C.sub.3)dialkyl, C.sub.1-C.sub.3alkylcarbonyloxy-, C.sub.1-C.sub.3alkoxycarbonyloxy-, C.sub.1-C.sub.3alkylcarbonylthio-, C.sub.1-C.sub.3alkylaminocarbonyl-, di(C.sub.1-C.sub.3)alkylaminocarbonyl-, fluorine or chlorine atom, and hydroxyl; or R.sup.40 and R.sup.41; R.sup.43 and R.sup.44; R.sup.49 and R.sup.50 are taken together to form a 5-membered or 6-membered ring, which ring may be 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)diaIkyl, 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 hydroxyl; f is an integer of from 0 to 2; with the proviso that when X does not comprise a C(O)O-motif with the carbonyl carbon in alpha or beta position to the oxygen atom of general formula (II), Y is an oxamide, a carbamate or a carbamide, preferably Y is an oxamide as defined above.

2. The compound according to claim 1, with the further proviso that when n is 3, 5, 6, 7 or 8, k is 1 and E is a group according to general formula (III) or general formula (IV), wherein each of R.sup.12 and R.sup.13 is a hydrogen atom; P represents a group:
(CH.sub.2).sub.3O(CH.sub.2)X.sup.81; (CH.sub.2).sub.5O(CH.sub.2)X.sup.81; wherein X.sup.81 represents a group selected from the group consisting of: ##STR00087## R.sup.1 is defined as R.sup.1 above; R.sup.2 represents NHR.sup.3; OR.sup.22; (OCH.sub.2CH.sub.2).sub.iR.sup.23; a mono-, or disaccharide, or a derivative thereof, which is joined to C(O) by an ester bond via the 1-O, 3-O, or 6-O-position of the saccharide; or wherein R.sup.2 is selected from the group consisting of: ##STR00088## 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 hydrogen or a C.sub.3-C.sub.6cycloalkyl group, which is optionally substituted with NH2, 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; R.sup.24, R.sup.25, R.sup.26, and R.sup.27 are as defined above; 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.8 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.

3. The compound according to claim 1, wherein X is ##STR00089## wherein R.sup.2 is OR.sup.22; (OCH.sub.2CH.sub.2).sub.iR.sup.23; a mono-, or disaccharide, or a derivative thereof, which is joined to C(O) by an ester bond via the 1-O, 3-O, or 6-O-position of the saccharide; or wherein R.sup.2 is selected from the group consisting of: ##STR00090## wherein R.sup.23 and i are as defined above; and wherein R.sup.22, and R.sup.23 to R.sup.27 are as defined in claim 1.

4. The compound according to claim 1, wherein X is C(O)OH or a suitable salt of the carboxylic acid, preferably a free carboxylic acid.

5. The compound according to claim 1, wherein Y is one of the oxamides defined according to claim 1.

6. The compound according to claim 1, wherein X is ##STR00091## wherein R.sup.2 is OR.sup.22; (OCH.sub.2CH.sub.2).sub.iR.sup.23; a mono-, or disaccharide, or a derivative thereof, which is joined to C(O) by an ester bond via the 1-O, 3-O, or 6-O-position of the saccharide; or wherein R.sup.2 is selected from the group consisting of: ##STR00092## wherein and R.sup.22, R.sup.23 to R.sup.27 and i are as defined in claim 1, and wherein Y is one of the oxamides defined according to claim 1.

7. The compound according to claim 1, wherein X is C(O)OH, preferably the free carboxylic acid, and Y is one of the oxamides defined according to claim 1.

8. The compound according to claim 1, with the formula (V): ##STR00093## wherein R.sup.55 represents OH; OR.sup.22; (OCH.sub.2CH.sub.2).sub.iR.sup.23; a mono-, or disaccharide, or a derivative thereof, which is joined to C(O) by an ester bond via the 1-O, 3-O, or 6-O-position of the saccharide; R.sup.22, R.sup.23 and i are as defined in claim 1, preferably R.sup.22 is a hydrogen atom or a C.sub.1-C.sub.6alkyl group, more preferably a hydrogen atom and i is preferably 2 to 4, more preferably 3; Y represents a group selected from the group consisting of: ##STR00094## wherein R.sup.40 to R.sup.50 are defined in claim 1, preferably R.sup.40 is a hydrogen atom or a C.sub.1-C.sub.6alkyl group, more preferably a hydrogen atom R.sup.57 and R.sup.58 are hydrogen; or form together a five- or six-membered ring, preferably an aromatic ring, optionally substituted with one to three or one to four 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; s is 1 or 2, with the proviso that s is 0 if R.sup.57 and R.sup.58 form together a five- or six-membered ring; the double bond in formula (V) represents a double carbon-carbon bond in cis-configuration, if R.sup.57 and R.sup.58 are hydrogen, or this double bond is part of a five- or six-membered ring formed together by R.sup.57 and R.sup.58.

9. The compound according to claim 8, wherein R.sup.55 represents OH or (OCH.sub.2CH.sub.2).sub.iR.sup.23; i is 2 to 4, preferably i is 3; R.sup.23 is preferably OH; Y is an oxamide, a carbamide or a carbamate, preferably a C.sub.1-C.sub.6alkyl substituted oxamide, carbamide or carbamate; R.sup.57 and R.sup.58 are both H, or together form a substituted or non-substituted five- or six-membered aromatic ring, preferably form a substituted or non-substituted benzyl ring; and s is 1 or s is 0 if R.sup.57 and R.sup.58 together form a substituted or non-substituted five- or six-membered aromatic ring.

10. The compounds according to claim 1, wherein the compound is selected from the group consisting of: ##STR00095## ##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100## or a pharmaceutically acceptable salt thereof.

11. The compound according to claim 1, with the formula (VI) ##STR00101## or a pharmaceutically acceptable salt thereof.

12. A pharmaceutical composition comprising the compound according to claim 1 in combination with a physiologically acceptable excipient.

13. The method according to claim 13, wherein the cardiovascular disease is selected from the group consisting of atrial fibrillation, ventricular arrhythmia, heart failure, coronary artery disease, myocardial infarction, maladaptive cardiac hypertrophy, and cardiac arrhythmias including ventricular extrasystoles, ventricular tachycardia, malignant ventricular tachycardia, atrial tachycardia, atrial flutter and atrial fibrillation, dilatative cardiomyopathy, and hypertensive heart disease, preferably selected from the group consisting of atrial fibrillation, atrial tachycardia, ventricular arrhythmia, heart failure.

14. The method according to claim 13, wherein the compound or composition is administered orally, topically, subcutaneously, intramuscularly, intraperitoneally, intravenously, intranasally.

15. The method according to claim 13, wherein the compound or composition is a dosage form selected from the group consisting of a spray, an aerosol, a foam, an inhalant, a powder, a tablet, a capsule, a soft gelatin capsule, a tea, a syrup, a granule, a chewable tablet, a salve, a cream, a gel, a suppository, a lozenge, a liposome composition and a solution suitable for injection.

16. (canceled)

17. A method for the treatment of a cardiovascular disease, comprising the step of administering a compound according to claim 1 to a subject in need thereof in an effective amount.

18. A method for the treatment of a cardiovascular disease, comprising the step of administering a composition according to claim 12, to a subject in need thereof in an effective amount.

Description

FIGURES

[0115] FIG. 1 is a column chart showing compounds 1 to 5 (Comp-01 to Comp-05), being examples of the invention, and other related structures (Comp-06 to Comp-13) with their potential to reduce spontaneous beating of NRCMs (For further details see Example 2 below).

[0116] FIG. 2 is a column chart showing a treatment with compound 2 (Comp-02), a synthetic agonist of 17,18-EEQ, which ameliorates the AF burden (A) and severity (B) of atrial fibrillation in a mouse model of moderate cardiac hypertrophy.

[0117] FIG. 3 is a column chart showing a treatment with compound 3 (Comp-03), a synthetic agonist of 17,18-EEQ, which ameliorates the duration (A) and severity (B) of cardiac arrhythmias in a rat model of myocardial infarction.

[0118] FIG. 4 is a column chart showing a treatment with compound 3 (Comp-03), a synthetic agonist of 17,18-EEQ, improving the post-ischemic functional recovery of isolated perfused mice hearts.

[0119] FIG. 5 is a column chart showing the inhibition of soluble epoxide hydrolase (sEH) by compounds of the invention (Comp-01 to Comp-03) in comparison to related analogs (Comp-07, Comp-08 and Comp-10).

[0120] FIG. 6 is a column chart showing the permeability potential of metabolically robust analogs of CYP-eicosanoids all part of the invention (Comp-01 to Comp-04) tested in Caco-2 cells.

[0121] FIG. 7 is a table summarizing data about the incorporation of compounds of the invention (Comp-02 and Comp-04) and other related compounds into membrane phospholipids.

[0122] FIG. 8 shows that continuous infusion of 100 nM of Comp-02 did not induce any obvious negative side effects. After global ischemia, contractility of the control hearts (n=5) was strongly reduced.

[0123] FIG. 9A to 9B show, that Comp-02 partially protected against OGD-induced damage in primary cardiomyocytes. FIG. 9C shows the LDH-release by Comp-02 and 17,18-EEQ.

EXAMPLE 1 SYNTHESIS OF COMPOUNDS

[0124] In the following the synthesis of selected compounds of the invention is illustrated.

Compound 1 (Comp-01)

[0125] Synthesis of compound 1 (Comp-01) was analogous to synthesis of compound 3 (Comp-03), while the urea-group was introduced following the synthetic route described in patent application WO2010/081683 (example 13).

Compound 2 (Comp-02)

Summary of Synthesis

[0126] ##STR00031## ##STR00032##

General Method

[0127] NMR spectra were recorded on Bruker Avance 400 MHz for .sup.1HNMR and 100 MHz for .sup.13CNMR. LCMS were taken on a quadrupole Mass Spectrometer on Shimadzu LCMS 2010 (Column: sepax ODS 502.0 mm, 5 um) or Agilent 1200 HPLC, 1956 MSD (Column: Shim-pack XR-ODS 303.0 mm, 2.2 um) operating in ES (+) ionization mode. Chromatographic purifications were by flash chromatography using 100200 mesh silica gel. Anhydrous solvents were pre-treated with 3A MS column before used. All commercially available reagents were used as received unless otherwise stated.

General Procedure for Preparation of Compound 2

[0128]

TABLE-US-00001 [00033] [00034] Reagent MW. amount mmol ratio Other Info. Compound 1 136.53 100 g 732.44 1 MeNH.sub.2HCl 67.52 64.29 g 952.17 1.3 Et.sub.3N 101.19 185.29 g 1830 2.5 THE 2 L Product 131.13 70 g 533.82 Yield: 73% (compound 2)

[0129] Methanamine (64.29 g, 952.17 mmol, 1.30 Eq) in 500 mL THF was added Et.sub.3N (75 g, 732.44 mmol), the solution was added to Compound 1 (100.00 g, 732.44 mmol, 1.00 eq), Et.sub.3N (111 g, 1.1 mol) in THF (1.5 L) at 10 C. And the mixture was stirred at 25 C. for 16 h. Then the mixture was filtered, the filtrate was washed with 2N HCl (500 mL), extracted with EA (300 mL*4), concentrated and purified by silica gel (PE: EA=3:1 to 1:1) to afford Compound 2 (70.00 g, 533.82 mmol, 72.88% yield) as a yellow oil.

[0130] TLC Information (PE: EtOAc=2:1); R.sub.f (Comp-02)=0.39; LCMS: ET2662-1-P1A (M+H.sup.+): 131.7; .sup.1H NMR (CDCl.sub.3, 400 MHz) 4.364.24 (q, J=8 Hz, 2H), 2.932.85 (d, J=4 Hz, 3H), 1.381.30 (t, J=8 Hz, 3H)

General Procedure for Preparation of Compound 4

[0131]

TABLE-US-00002 [00035] [00036] Reagent MW. amount mmol ratio Other Info. Compound 3 98.14 47.5 g 484 1 Compound 4 147.13- 78.33 g 532.4 1.1 PPh.sub.3 262.29 133.3 g 508.2 1.05 DIAD 202.21 107.66 g 532.4 1.1 THF 1.8 L Product 227.26 42.5 g 374.02 Yield 77.3% (compound 5)

[0132] A solution of Compound 3 (47.50 g, 484.00 mmol, 1.00 eq.) and DIAD (107.66 g, 532.40 mmol, 1.10 eq.) in anhydrous THF (50 mL) was slowly added via cannula to a 0 C. solution of Compound 4 (78.33 g, 532.40 mmol, 1.10 eq.) and PPh.sub.3 (133.30 g, 508.20 mmol, 1.05 eq.) in anhydrous THF (100 mL). The flask and cannula were washed with an additional portion of dry THF (30 mL) to ensure complete addition. The reaction was allowed to gradually warm to 25 C. and stirred for 18 h. Then H.sub.2O (1000 mL) was added, extracted with EA (500 mL*2), concentrated and purified by silica gel (PE:EA=0-10:1) to give Compound 5 (42.5 g, 374.02 mmol, 77.28% yield) as a white solid.

[0133] TLC Information (PE: EtOAc=5:1); R.sub.f (Comp-03)=0.2; R.sub.f (Comp-05)=0.5; .sup.1H NMR (CDCl.sub.3, 400 MHz) 7.867.79 (m, 2H), 7.727.67 (m, 2H), 3.733.66 (t, J=8 Hz, 2H), 2.272.20 (m, 2H), 1.951.91 (t, J=4 Hz, 1H), 1.851.75 (m, 2H), 1.611.52 (m, 2H)

General Procedure for Preparation of Compound 6

[0134]

TABLE-US-00003 [00037] [00038] Reagent MW. amount mmol ratio Other Info. Compound 5 227.26 88 g 387.22 1 NIS 224.98 130.68 g 580.83 1.5 AgNO.sub.3 169.87 16.44 g 96.81 0.25 THF 1.6 L Product (Compound 6) 353.15 118.6 g 335.8 Yield: 86%

[0135] NIS (130.68 g, 580.83 mmol, 1.50 eq.) was added in one portion to a solution of Compound 5 (88.00 g, 387.22 mmol, 1.00 eq.) and AgNO.sub.3 (16.44 g, 96.81 mmol, 0.25 eq.) in anhydrous THF (1600 mL) at 25 C. The reaction head space was flushed with N.sub.2 and the reaction mixture was protected from light with an aluminum foil wrap and stirred for 16 h. The mixture was poured into water (1000 mL), extracted with EA (600 mL*3), concentrated and purified by silica (PE: EA=10:1 to 2:1) to give Compound 6 (118.6 g, 1.01 mol, 86.78% yield) as a white solid.

[0136] TLC Information (PE: EtOAc=20:1); R.sub.f (Comp-05)=0.22; R.sub.f(Cpd 6)=0.21; .sup.1H NMR (CDCl.sub.3, 400 MHz) 7.877.82 (m, 2H), 7.747.69 (m, 2H), 3.743.67 (t, J=8 Hz, 2H), 2.452.39 (t, J=8 Hz, 2H), 1.841.74 (m, 2H), 1.611.52 (m, 2H)

General Procedure for Preparation of Compound 7

[0137]

TABLE-US-00004 [00039] [00040] Reagent MW. amount mmol ratio Other Info. Compound 6 353.15 157 g 444.57 1 BH.sub.3.DMS 58 mL 577.94 1.3 2-methyl-2-butene 70.13 87.3 g 1240 2.8 AcOH 260 mL THF 1.2 L Product (compound 5) 355.17 135 g 380.1 Yield: 85%

[0138] 2-methylbut-2-ene (87.30 g, 1.24 mol, 2.80 eq.) was added over 30 min to a 0 C. solution of BH3.Me2S (43.91 g, 577.94 mmol, 1.30 eq.) in THF (300 mL). After 1 h, the reaction mixture was warmed to 25 C. and stirred for 90 min. After re-cooling to 0 C., a solution of Compound 6 (157.00 g, 444.57 mmol, 1.00 eq.) in THF (900 mL) was added slowly over 1 h. Upon complete addition, the cold bath was removed and the reaction mixture was stirred at 25 C. After 2 h, the reaction was cooled again to 0 C. where upon glacial AcOH (260 mL) was added slowly over 30 min (Caution: gas evolution) and stirred at 25 C. for 16 h. TLC (PE: EA=10:1) show the reaction was completed, the mixture was pour into water (1 L), extracted with EA (300 mL*2), concentrated and purified by silica gel (PE:EA=0-10:1) to give Compound 7 (135 g, 380.1 mmol, 85.50% yield) as a yellow oil.

[0139] TLC Information (PE: EtOAc=10:1); R.sub.f (Cpd 6)=0.5; R.sub.f(Cpd 7)=0.55; .sup.1H NMR: (CDCl.sub.3, 400 MHz) 7.887.80 (m, 2H), 7.757.67 (m, 2H), 6.246.11 (m, 2H), 3.743.66 (t, J=8 Hz, 2H), 2.242.15 (q, J=8 Hz, 2H), 1.781.67 (m, 2H), 1.551.44 (m, 2H)

General Procedure for Preparation of Compound 8

[0140]

TABLE-US-00005 [00041] [00042] Reagent MW. amount mmol ratio Other Info. Compound 7 355.17 138 g 388.55 1 N.sub.2H.sub.4H.sub.2O 50.06 97.25 g 1940 5 MeOH 2 L Product (compound 8) 225.07 81 g 683.79 Yield: 92%

[0141] N.sub.2H.sub.4.H.sub.2O (97.25 g, 1.94 mol, 5.00 eq.) was added to a solution of Compound 7 (138.00 g, 388.55 mmol, 1.00 eq) in anhydrous MeOH (2.00 L) at 0 C. and stirred at 25 C. for 18 h, TLC (PE:EA=10:1) show the reaction was completed, the reaction mixture was concentrated, the residue was poured into DCM (5000 mL) and stirred for 30 mins. Filtered and the filter cake was washed with DCM (1 L*2), the filtrate was concentrated to give Compound 8 (162.00 g, crude) as a yellow oil.

[0142] TLC Information (PE: EtOAc=10:1); R.sub.f (Cpd 7)=0.5; R.sub.f(Cpd 8)=0; TLC Information (DCM: MeOH=10:1); R.sub.f (Cpd 7)=1; R.sub.f (Cpd 8)=0.2; .sup.1H NMR: (CDCl.sub.3, 400 MHz) 6.196.07 (m, 2H), 2.732.59 (m, 2H), 2.202.05 (m, 2H), 1.751.55 (m, 2H), 1.511.36 (m, 4H)

General Procedure for Preparation of Compound 9

[0143]

TABLE-US-00006 [00043] [00044] Reagent MW. amount mmol ratio Other Info. Compound 8 225.07 92 g 408.76 1 Compound 2 131.13 53.6 g 408.76 1 Et3N 101.19 49.64 g 590.51 1.2 EtOH 1.5 L Product (compound 9) 310.13 90 g 232.16 Yield: 57%

[0144] Compound 8 (92.00 g, 408.76 mmol, 1.00 eq) Compound 2 (53.60 g, 408.76 mmol, 1.00 eq) and Et.sub.3N (49.64 g, 490.51 mmol, 1.20 eq) in anhydrous ethanol (1.5 L) was heated at 60 C. for 20 h. TLC (DCM:MeOH=10:1) show the reaction was completed, the mixture was concentrated to about 300 mL. Filtered and concentrated to give Compound 9 (90 g, 232.16 mmol, 57% yield) as a white solid.

[0145] TLC Information (DCM: MeOH=10:1); R.sub.f (Cpd 8)=0.2; R.sub.f (Cpd 9)=0.5; .sup.1H NMR: (CDCl.sub.3, 400 MHz) 7.577.37 (s, 2H), 6.256.20 (d, J=8 Hz, 1H), 6.186.11 (q, J=8 Hz, 1H), 3.37-3.30 (q, J=8 Hz, 2H), 2.932.88 (d, J=4 Hz, 3H), 2.212.13 (m, 2H), 1.661.56 (m, 2H), 1.531.43 (m, 2H)

General Procedure for Preparation of Compound 12

[0146]

TABLE-US-00007 [00045] [00046] Reagent MW. amount mmol ratio Other Info. Compound 10 114.1 25 g 197.2 1 90% Compound 11 114.18 27.02 g 236.63 1.2 In(OTf).sub.3 560 22.09 g 39.44 0.2 toluene 350 mL Compound 12 200.27 35 g 139.81 Yield: 71%

[0147] Compound 10 (25.00 g, 197.20 mmol, 1.00 eq.) in toluene (75 mL) was added to a solution of Compound 11 (27.02 g, 236.63 mmol, 1.20 eq.) In (OTf).sub.3 (22.09 g, 39.44 mmol, 0.20 eq) in toluene (275 mL) over 20 mins. Then the mixture was stirred at 25 C. for 48 h. The mixture was concentrated and purified by silica gel (PE: EA=20:1) to give ethyl Compound 12 (35.00 g, 139.81 mmol, 70.90% yield, 80% purity) as a yellow oil.

[0148] TLC Information (PE: EtOAc=10:1); R.sub.f (Cpd 11)=0.21; R.sub.f(Cpd 12)=0.55; .sup.1H NMR: (CDCl.sub.3, 400 MHz) 5.865.72 (m, 1H), 5.035.86 (m, 2H), 4.244.17 (q, J=8 Hz, 2H), 4.074.01 (s, 2H), 3.543.47 (t, J=8 Hz, 2H), 2.091.98 (m, 2H), 1.681.55 (m, 2H), 1.451.32 (m, 4H), 1.301.25 (t, J=8 Hz, 3H)

General Procedure for Preparation of Compound 13

[0149]

TABLE-US-00008 [00047] [00048] Reagent MW. amount mmol ratio Other Info. Compound 12 200.27 10.07 g 50.3 1.2 90% Compound 9 310.13 13 g 41.92 1 9-BBN 100.6 mL 100.6 2.4 Na.sub.2CO.sub.3 200 mL 2M Pd(PPh.sub.3)Cl.sub.2 701.9 1.47 g 2.1 0.05 THF 800 mL Compound 13 384.51 6.5 g 16.06 Yield: 38%

[0150] To an oven-dried flask containing 9-BBN (17.53 g, 100.60 mmol, 2.40 eq) in THF (540 mL) was added a solution of Compound 12 (10.07 g, 50.30 mmol, 1.20 eq) in THF (60 mL) at 0 C. After stirring at 25 C. for 16 h, an aqueous solution of Na.sub.2CO.sub.3 (200 mL of 2 M soln prepared from argon sparged H.sub.2O) was added. After 2 h, Pd(PPh.sub.3).sub.2Cl.sub.2 (1.47 g, 2.10 mmol, 0.05 eq) was added followed by Compound 9 (13.00 g, 41.92 mmol, 1.00 eq) dissolved in THF (200 mL). The resulting red solution was protected from light. The reaction was stirred at 50 C. for 5 h. LCMS show the reaction was completed. After cooling to 25 C., the reaction mixture was concentrated in vacuo and the residue was purified by silica gel (PE:EA=10:1 to 3:1) to give Compound 13 (6.5 g, 16.06 mmol, 38.31% yield, 95% purity) as a yellow solid.

[0151] TLC Information (PE: EtOAc=2:1); R.sub.f (Cpd 12)=0.3; R.sub.f(Cpd 13)=0.3; LCMS: ET2662-38-P1D (M+H.sup.+): 385.1; .sup.1H NMR: (CDCl.sub.3, 400 MHz) 7.577.38 (s, 1H), 5.415.25 (m, 2H), 4.254.17 (q, J=8 Hz, 2H), 4.074.02 (s, 2H), 3.543.47 (t, J=8 Hz, 2H), 3.343.26 (q, J=8 Hz, 2H), 2.922.87 (d, J=8 Hz, 3H), 2.081.94 (m, 4H), 1.651.51 (m, 4H), 1.431.23 (m, 13H)

General Procedure for Preparation of Comp-02

[0152]

TABLE-US-00009 [00049] [00050] Reagent MW. amount mmol ratio Other Info. Compound 13 384.51 7.5 g 19.51 1 90% LiOH 23.95 0.9341 g 39.02 2 H.sub.2O 40 mL THF 70 mL Compound 13 356.46 4 g 10.72 Yield: 55%

[0153] To a solution of Compound 13 (7.50 g, 19.51 mmol, 1.00 eq.) in THF (70.00 mL) was added LiOH (934.31 mg, 39.02 mmol, 2.00 eq.) in H.sub.2O (40.00 mL) at 0 C. and then the reaction mixture was stirred at 0-25 C. for 1 h. LCMS show the reaction was completed. Then H.sub.2O (60 mL) was added to the reaction mixture, the aqueous phase was treated with 3 N HCl (10 mL) to pH=3-4, extracted with EA (100 mL*3), dried, the organic phase was concentrated to give crude product. The residue was purified by column on gel (PE: EA=5:1 to EA) to give Comp-02 (4.00 g, 10.72 mmol, 54.95% yield, 95.51% purity)

[0154] TLC Information (DCM: MeOH=10:1); R.sub.f (Cpd 13)=0.9; R.sub.f (Comp-02)=0.4; MS: ET2662-43-P1C (M+Na.sup.+): 379.2; .sup.1H NMR (CDCl.sub.3, 400 MHz) 7.84 (s, 1H), 7.74 (s, 1H), 5.405.32 (m, 2H), 4.11 (s, 2H), 3.593.55 (t, J=6.4 Hz, 2H), 3.353.32 (t, J=6.8 Hz, 2H), 2.922.91 (d, J=5.2 Hz, 3H), 2.072.00 (m, 4H), 1.64-1.59 (m, 4H), 1.421.32 (m, 10H); .sup.13C NMR (CDCl3, 100 MHz) 173.7, 160.7, 159.8, 130.5, 129.0, 72.0, 67.8, 39.7, 29.4, 29.3, 29.0, 29.0, 28.6, 27.1, 26.8, 26.7, 25.8

Compound 3 (Comp-03)

Summary of Synthesis

[0155] ##STR00051## ##STR00052##

Synthesis of 2-(hex-5-yn-1-yl)isoindoline-1,3-dione (2)

[0156] Following literature precedent,.sup.1 a solution of 5-hexyn-1-ol (1) (5 g, 1 equiv) and diisopropyl azodicarboxylate (DIAD, 10.5 g, 1.02 equiv) in anhydrous THF (30 mL) was slowly added via cannula to a 0 C. solution of phthalimide (7.5 g, 51 mmol) and triphenylphosphine (TPP, 13.4 g, 1 equiv) in anhydrous THF (50 mL). The flask and cannula were washed with an additional portion of dry THF (20 mL) to ensure complete addition. The reaction was allowed to gradually warm to room temperature overnight. After a total of 18 h, all volatiles were evaporated and the residue was purified using a Teledyne Isco Combiflash RF chromatographic system (80 g SiO.sub.2 column eluted with hexanes, 2 min; 0-20% EtOAc/hexanes, 12 min; 20% EtOAc/hexanes, 6 min) to give 2 (8.3 g, 72%) as a white solid whose spectral values were identical to those reported..sup.2

##STR00053##

Synthesis of 2-(6-iodohex-5-yn-1-yl)isoindoline-1,3-dione (3)

[0157] Following literature precedent,.sup.3 N-iodosuccinimide (NIS, 7.4 g, 1.5 equiv) was added in one portion to a rt solution of alkyne 2 (5.0 g, 22 mmol) and AgNO.sub.3 (0.93 mg, 0.25 equiv) in anhydrous THF (120 mL). The reaction head space was flushed with argon and the reaction mixture was protected from light with an aluminum foil wrap. After 4 h, the reaction mixture was poured into H.sub.2O (200 mL) and extracted with Et.sub.2O (250 mL). The ethereal extracts were washed with brine (360 mL) (Note: biphasic mixture turned brown). The combined aqueous phases were re-extracted with Et.sub.2O (250 mL). The combined ethereal extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated on a rotary evaporator. The residue was purified using a Teledyne Isco Combiflash RF chromatographic system (80 g SiO.sub.2 column eluted with hexanes, 2 min; 0-40% EtOAc/hexanes, 8 min; 40% EtOAc/hexanes, 10 min; 40-100% EtOAc/hexanes, 5 min; 100%, EtOAc, 3 min) to give 3 (97%) as a white solid, mp 132.5-132.7 C. .sup.1H NMR (500 MHz, CDCl.sub.3) 7.85 (ddd, J=5.4, 3.0, 1.0 Hz, 2H), 7.72 (ddd, J=5.5, 3.0, 1.0 Hz, 2H), 3.71 (t, J=7.1 Hz, 2H), 2.42 (t, J=7.0 Hz, 2H), 1.83-1.73 (m, 2H), 1.61-1.51 (m, 2H); .sup.13C NMR (100 MHz, CDCl.sub.3) 168.62, 134.14, 132.30, 123.44, 94.04, 37.60, 27.91, 25.89, 20.60, 6.27.

##STR00054##

Synthesis of 2-(6-iodohex-5(Z)-en-1-yl)isoindoline-1,3-dione (4)

[0158] Following literature precedent,.sup.4 neat 2-methyl-2-butene (4.2 mL, 2.8 equiv) was added over 5 min to a 0 C. solution of BH.sub.3.Me.sub.2S (2.0 M in THF, 9.2 mL, 1.3 equiv) in THF (3 mL). After 1 h, the reaction mixture was warmed to room temperature and stirred for 90 min. After re-cooling to 0 C., a solution of iodoalkyne 3 (5 g, 1 equiv) in THF (30 mL) was added slowly over 5 min. Upon complete addition, the cold bath was removed and the reaction mixture was stirred at rt. After 2 h, the reaction was cooled again to 0 C. whereupon glacial AcOH (8.5 mL) was added slowly over 5 min (Caution: gas evolution). After stirring overnight (14 h), the reaction mixture was diluted with H.sub.2O (20 mL), then carefully poured into a stirring, saturated sodium bicarbonate solution (40 mL). The biphasic mixture was extracted with ether (240 mL) and the combined ethereal extracts were washed with water, brine, dried over anhydrous MgSO.sub.4, filtered, and concentrated in vacuo. The residue was purified using a Teledyne Isco Combiflash RF chromatographic system (40 g SiO.sub.2 column eluted with 0-20% EtOAc/hexanes, 8 min; 20% EtOAc/hexanes, 6 min) to give a mixture (4.52 g) of 4 and borane side-product. Further purification was postponed until the next step.

##STR00055##

Synthesis of 6-iodohex-5(Z)-en-1-amine (5)

[0159] Following literature precedent,.sup.5 40% wt MeNH.sub.2 in H.sub.2O (15 mL) was added to a rt solution of crude 4 (4.52 g) in anhydrous EtOH (20 mL). After stirring overnight (18 h), the reaction mixture was poured into ice water (100 mL) and extracted with Et.sub.2O (30 mL2). The combined ethereal extracts were washed with cold 1N HCl solution (20 mL2). The combined aqueous washes were adjusted to pH 8 with dilute, aq. NaOH. The solution was extracted with Et.sub.2O (30 mL2), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to give crude 5 (1.12 g) as a brown oil that was used in the next step without further purification. .sup.1H NMR (500 MHz, CDCl.sub.3) 6.29-6.08 (m, 2H), 2.71 (tt, J=7.0, 1.8 Hz, 2H), 2.16 (app q, J=6.5 Hz, 2H), 1.78-1.52 (m, 2H).

##STR00056##

Synthesis of N.SUP.1.-(6-iodohex-5(Z)-en-1-yl)-N.SUP.2.-methyloxalamide (7)

[0160] Following literature precedent,.sup.6 a solution of iodoalkene 5 (1.12 g, 4.98 mmol), ethyl 2-(methylamino)-2-oxoacetate (6) (0.62 g, 1.2 equiv) and triethylamine (0.83 mL, 1.2 equiv) in anhydrous ethanol (10 mL) was heated at 60 C. After 20 h, the brown solution was cooled to rt and concentrated in vacuo. Purification of the residue using a Teledyne Isco Combiflash RF chromatographic system (25 g SiO.sub.2 column eluted with 0-50% EtOAc/hexanes, 10 min; 50% EtOAc/hexanes, 10 min) gave 7 (0.93 g, 60%) as a white solid, 99.7-99.8 C. .sup.1H NMR (500 MHz, CDCl.sub.3) 7.46 (br s, 2H), 6.32-6.02 (m, 2H), 3.34 (app q, J=6.9 Hz, 2H), 2.91 (d, J=5.3 Hz, 3H), 2.18 (dt, J=7.5, 7.0 Hz, 2H), 1.68-1.59 (m, 2H), 1.54-1.42 (m, 2H);.sup.13C NMR (100 MHz, CDCl.sub.3) 160.47, 159.70, 140.43, 83.07, 39.40, 34.11, 28.61, 26.15, 25.11.

##STR00057##

Synthesis of ethyl 2-(oct-7-en-1-yloxy)acetate (10)

[0161] Following literature precedent,.sup.7 neat 8 (1.92 g, 1.2 equiv) was added to a rt suspension of In(OTf).sub.3 (1.57 g, 20 mol %) in anhydrous toluene (20 mL). Ethyl diazoacetate (9) (1.60 g, 14 mmol) was added slowly under an argon atmosphere over 5 min (caution: exothermic) to give a yellow solution. After 2 days, the reaction mixture was concentrated in vacuo and the residue was purified using a Teledyne Isco Combiflash RF chromatographic system (25 g SiO.sub.2 column eluted with 0-10% EtOAc/hexanes, 5 min; 10% EtOAc/hexanes, 8 min) to give 10 (2.72 g, 97%) as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3) 5.80 (ddt, J=16.9, 10.2, 6.6 Hz, 1H), 5.08-4.84 (m, 2H), 4.22 (q, J=7.1 Hz, 2H), 4.06 (s, 2H), 3.52 (t, J=6.7 Hz, 2H), 2.13-1.96 (m, 2H), 1.72-1.52 (m, 2H), 1.48-1.33 (m, 4H), 1.28 (t, J=7.1 Hz, 3H);.sup.13C NMR (100 MHz, CDCl.sub.3) 170.70, 138.99, 114.48, 71.97, 68.48, 60.86, 33.84, 29.55, 28.84, 25.63, 14.34.

##STR00058##

Synthesis of ethyl 2-((13-(2-(methylamino)-2-oxoacetamido)tridec-8(Z)-en-1-yl)oxy)acetate (11)

[0162] To an oven-dried flask containing ethyl 2-(oct-7-en-1-yloxy)acetate (10) (220 mg, 1.2 equiv) was added a solution of 9-BBN (0.5 M in THF, 2.4 equiv, 4.40 mL). After stirring at rt for 3 h, an aqueous solution of Na.sub.2CO.sub.3 (1.5 mL of 2 M soln prepared from argon sparged H.sub.2O) was added. After 5 min, Pd(PPh.sub.3).sub.2Cl.sub.2 (33 mg, 5 mol %) was added followed by 7 (284 mg, 0.92 mmol) dissolved in THF (4 mL). The resulting red solution was protected from light while another portion of aq. Na.sub.2CO.sub.3 (0.5 mL of 2 M soln) was added. The reaction was continued overnight (14 h) at rt, then at 50 C. for 4 h. After cooling to rt, the reaction mixture was concentrated in vacuo and the residue was purified using a Teledyne Isco Combiflash RF chromatographic system (24 g SiO.sub.2 column eluted with 0-40% EtOAc/hexanes, 6 min; 40% EtOAc/hexanes, 8 min; 40-100% EtOAc/hexanes, 4 min) to give ether 11 (330 mg, 90%) as an off-white solid. An analytical sample was purified by preparative TLC to give 11 as a white low melting solid.

[0163] TLC: 50% EtOAc/hexanes, R.sub.f0.49. .sup.1H NMR (500 MHz, CDCl.sub.3) 7.45 (br s, 2H), 5.42-5.26 (m, 2H), 4.22 (q, J=7.2 Hz, 2H), 4.06 (s, 2H), 3.52 (t, J=6.7 Hz, 2H), 3.31 (dt, J=7.0, 6.5 Hz, 2H), 2.91 (d, J=5.1 Hz, 3H), 2.15-1.91 (m, 4H), 1.70-1.50 (m, 2H), 1.44-1.31 (m, 12H), 1.29 (t, J=7.1 Hz, 3H);.sup.13C NMR (100 MHz, CDCl.sub.3) 170.62, 160.55, 159.66, 130.58, 128.86, 71.96, 68.64, 39.55, 29.61, 29.51, 29.30, 29.19, 27.20, 26.83, 26.67, 26.15, 25.93, 14.21.

##STR00059##

Synthesis of 2-((13-(2-(methylamino)-2-oxoacetamido)tridec-8(Z)-en-1-yl)oxy)acetic acid (12)

[0164] To a rt solution of 11 (720 mg, 1.87 mmol) in THF (44 mL) was added LiOH (9 mL of 1.0 M aq. solution). After 48 h, the reaction was cooled to 4 C. and acidified to pH 4 using aq. 2 N HCl. The mixture was diluted with H.sub.2O (10 mL) and extracted with EtOAc (15 mL3). The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered through a fritted funnel, and concentrated in vacuo. The crude material was purified using a Teledyne Isco Combiflash RF chromatographic system (12 g SiO.sub.2 column eluted with 0-80% EtOAc/hexanes, 15 min; 80% EtOAc/hexanes, 5 min) to give 12 (232 mg, 33%) as a white solid, mp 94.6-94.7 C.

[0165] .sup.1H NMR (500 MHz, CDCl.sub.3) 7.90 (s, 1H), 7.66 (s, 1H), 5.48-5.22 (m, 2H), 4.10 (s, 2H), 3.58 (t, J=6.5 Hz, 2H), 3.32 (dt, J=7.0, 6.5 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.16-1.90 (m, 4H), 1.71-1.48 (m, 4H), 1.45-1.18 (m, 10H); .sup.13C NMR (75 MHz, CD.sub.3OD) 176.96, 160.32, 160.12, 130.65, 129.99, 72.51, 69.84, 39.45, 29.82, 29.58, 29.15, 27.71, 27.38, 27.24, 27.08, 26.83, 25.84, 25.03.

##STR00060##

[0166] Synthesis of Comp-03:

[0167] A mixture of EDCI (275 mg, 1.3 equiv) and triethyleneglycol (1.5 mL, 10 equiv) was dried under high vacuum for 90 min. The reaction flask was flushed with argon and DMAP (175 mg, 1.3 equiv), acetonitrile (50 mL), and acid 12 (395 mg, 1.1 mmol) dissolved in CH.sub.2Cl.sub.2 (20 mL) were added. After 3 days, the reaction mixture was concentrated in vacuo, the crude residue was dissolved in EtOAc (20 mL) and washed with 1N HCl (20 mL) and brine (20 mL). The aqueous washings were re-extracted with EtOAc (20 mL2). The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The residue was purified using a Teledyne Isco Combiflash RF chromatographic system (12 g SiO.sub.2 column eluted with 0-80% EtOAc/hexanes, 8 min; 80% EtOAc/hexanes, 4 min; 80-100% EtOAc/hexanes, 3 min; 100% EtOAc, 15 min; 10% MeOH/CH.sub.2Cl.sub.2, 5 min) to give analog 13 (174 mg, 32%) as a white solid, mp 65.3-65.8 C.

[0168] .sup.1H NMR (500 MHz, CDCl.sub.3) 7.46 (s, 2H), 5.41-5.27 (m, 2H), 4.33 (t, J=4.7 Hz, 2H), 4.11 (s, 2H), 3.77-3.70 (m, 4H), 3.70-3.64 (m, 4H), 3.61 (app t, J=4.5 Hz, 2H), 3.52 (t, J=6.7 Hz, 2H), 3.42 (t, J=6.1 Hz, OH), 3.31 (dt, J=7.0, 6.5 Hz, 2H), 2.91 (d, J=5.2 Hz, 3H), 2.44 (s, 1H), 2.05 (dt, J=7.5, 7.0 Hz, 2H), 2.00 (dt, J=7.0, 6.5 Hz, 2H), 1.62-1.50 (m, 4H), 1.45-1.21 (m, 10H); .sup.13C NMR (125 MHz, CDCl.sub.3) 170.86, 160.83, 159.95, 130.76, 129.12, 72.76, 72.21, 70.77, 70.52, 69.19, 68.34, 63.84, 61.92, 39.78, 29.84, 29.73, 29.54, 29.42, 29.02, 27.44, 27.09, 26.92, 26.42, 26.15.

Compound 4 (Comp-04)

[0169] Synthesis of Compound 4 (Comp-04) was analogous to synthesis of compound 2 (Comp-02), while the urea-group was introduced following the synthetic route described in patent application WO2010/081683 (example 6).

Compound 5 (Comp-05)

Summary of Synthesis

[0170] ##STR00061##

General Procedure for Preparation of Compound 4-2

[0171]

TABLE-US-00010 [00062] [00063] Reagent MW. amount mol ratio Other Info. Cpd. 4-1 219.02 30.0 g 137 mmol 1.0 eq. Cpd. 1 98.14 13.4 g 137 mmol 1.0 eq. CuI 190.45 522 mg 2.74 mmol 0.02 eq. Pd(PPh.sub.3).sub.4 1155.56 1.58 g 1.37 mmol 0.01 eq. TEA 480 mL Product: (Cpd. 4-2) 189.25 21.0 g 99.9 mmol Yield: 73%

[0172] A mixture of Cpd.4-1 (30.0 g, 137 mmol, 1.0 eq) in TEA (480 mL) was added Cpd.1 (13.4 g, 137 mmol, 1.0 eq), CuI (522 mg, 2.74 mmol, 0.02 eq), Pd(PPh.sub.3).sub.4 (1.58 g, 1.37 mmol, 0.01 eq) under N.sub.2 at 25 C. and stirred at 25 C. for 16 hrs. TLC (petroleum ether/ethyl acetate=1/1, R.sub.f=0.5) showed that the reaction was complete. The solution was poured into aq.NH.sub.4Cl (1.0 L), extracted with DCM (200 mL*5), the combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel eluted with petroleum ether: EtOAc (10:1, 1:1) to give Cpd.4-2 (21.0 g, 73% yield) as yellow oil.

[0173] .sup.1H NMR: ET5008-6-P1b1 400 MHz CDCl.sub.3; 7.30-7.24 (m, 1H), 7.10 (t, J=7.6 Hz, 1H), 6.73-6.65 (m, 2H), 4.19 (br, 2H), 3.74 (m, 2H), 2.54 (t, J=6.0 Hz, 2H), 1.87-1.68 (m, 4H), 1.50-1.45 (m, 1H).

General Procedure for Preparation of Cpd.4-3

[0174]

TABLE-US-00011 [00064] [00065] Reagent MW. amount mol ratio Other Info. Cpd. 4-2 189.25 21.0 g 111 mmol 1.0 eq. Pd/C 500 mg MeOH 500 mL Product: (Cpd. 4-3) 193.28 17.0 g 83.6 mmol Yield: 75%

[0175] A mixture of Cpd.4-2 (21.0 g, 111 mmol, 1.0 eq) in MeOH (500 mL) was added Pd/C (500 mg) and stirred at 25 C. under 50 psi of H.sub.2 for 16 hrs. LC-MS (ET5008-10-P1A5, product: RT=1.10 min) show that the reaction was complete. Then the solution was filtered and concentrated to give Cpd.4-3 (17.0 g, 75% yield) as yellow oil.

[0176] .sup.1H NMR: ET5008-10-P1b1 400 MHz CDCl.sub.3; 7.08-7.03 (m, 2H), 6.78-6.69 (m, 2H), 3.69-3.62 (m, 4H), 2.52 (t, J=8.0 Hz, 2H), 1.68-1.59 (m, 4H), 1.47-1.42 (m, 4H), 1.31-1.27 (m, 1H).

General Procedure for Preparation of Cpd.4-4

[0177]

TABLE-US-00012 [00066] [00067] Reagent MW. amount mol ratio Other Info. Cpd. 4-3 193.28 17.0 g 88.0 mmol 1.0 eq. NaNO.sub.2 69.00 6.07 g 88.0 mmol 1.0 eq. KI 166.00 43.8 g 264 mmol 3.0 eq. ConH.sub.2SO.sub.4 98.08 30.2 g 308 mmol 3.5 eq. H.sub.2O 560 mL Product: (Cpd. 4-4) 304.17 17.0 g 50.3 mmol Yield: 57%

[0178] Con.H.sub.2SO.sub.4 (30.2 g, 308 mmol, 3.5 eq) was added to Cpd.4-3 (17.0 g, 88.0 mmol, 1.0 eq) in H.sub.2O (500 mL) at 0 C. under N.sub.2. A solution of NaNO.sub.2 (6.07 g, 88.0 mmol, 1.0 eq) in H.sub.2O (30.0 mL) was added to the solution at 0 C. and stirred at 0 C. for 15 mins. A solution of KI (43.8 g, 264 mmol, 3.0 eq) in H.sub.2O (30.0 mL) was added at 0 C. and the resulting suspension was warmed to 25 C. and stirred for 45 mins. TLC (petroleum ether/ethyl acetate=1/1, R.sub.f=0.9) showed that the reaction was complete. H.sub.2O (400 mL) was added, extracted with EtOAc (350 mL*3), the combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel eluted with petroleum ether: EtOAc (100:1, 10:1) to give Cpd.4-4 (17.0 g, 57% yield) as brown oil.

[0179] .sup.1H NMR: ET5008-22-P1b1 400 MHz CDCl.sub.3; 7.80 (d, J=7.2 Hz, 1H), 7.28-7.23 (m, 1H), 7.21-7.18 (m, 1H), 6.89-6.85 (m, 1H), 3.65 (t, J=6.8 Hz, 2H), 2.71 (t, J=8.0 Hz, 2H), 1.61-1.50 (m, 4H), 1.45-1.40 (m, 4H), 1.31-1.28 (m, 1H).

General Procedure for Preparation of Compound 4-5

[0180]

TABLE-US-00013 [00068] [00069] Reagent MW. amount mol ratio Other Info. Cpd. 4-4 304.17 10.0 g 32.9 mmol 1.0 eq. BrCH.sub.2CO.sub.2tBu 195.05 7.70 g 39.5 mmol 1.2 eq. KOH 56.11 33.0 g 588 mmol 18 eq. Bu.sub.4NHSO.sub.4 339.53 5.58 g 16.4 mmol 0.50 eq. toluene 50.0 mL H.sub.2O 50.0 mL Product: (Cpd. 4-5) 418.31 5.40 g 12.3 mmol Yield: 37%

[0181] A mixture of BrCH.sub.2CO.sub.2tBu (7.70 g, 39.5 mmol, 1.2 eq) and Cpd.4-4 (10.0 g, 32.9 mmol, 1.0 eq) in toluene (50.0 mL) was added Bu.sub.4NHSO.sub.4 (5.58 g, 16.4 mmol, 0.50 eq), KOH (33.0 g, 588 mmol, 17.9 eq) in H.sub.2O (50.0 mL) at 0 C., then the mixture was stirred at 25 C. for 16 hrs. TLC (petroleum ether/ethyl acetate=10/1, R.sub.f=0.62) show 40% SM remained. H.sub.2O (200 mL) was added, extracted with DCM (200 mL*2), the combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel eluted with petroleum ether: EtOAc (40:1) to give Cpd.4-5 (5.40 g, 37% yield) as yellow oil.

[0182] .sup.1H NMR: ET5008-26-P1b1 400 MHz CDCl.sub.3; 7.82 (d, J=7.2 Hz, 1H), 7.30-7.26 (m, 1H), 7.23-7.20 (m, 1H), 6.91-6.88 (m, 1H), 3.97 (s, 2H), 3.53 (t, J=6.8 Hz, 2H), 2.72 (t, J=8.0 Hz, 2H), 1.69-1.59 (m, 4H), 1.58-1.43 (m, 13H).

General Procedure for Preparation of Cpd.4-6

[0183]

TABLE-US-00014 [00070] [00071] Reagent MW. amount mol ratio Other Info. Cpd. 4-5 418.31 5.40 g 12.9 mmol 1.0 eq. Cpd. 2 169.22 2.18 g 12.9 mmol 1.0 eq. CuI 190.45 49.2 mg 258 umol 0.02 eq. PdCl.sub.2(PPh.sub.3).sub.2 701.90 181 mg 258 umol 0.02 eq. Et.sub.3N 110 mL Product: (Cpd. 4-6) 459.62 3.00 g 6.20 mmol Yield: 48%

[0184] A mixture of Cpd.4-5 (5.40 g, 12.9 mmol, 1.0 eq) and Cpd.2 (2.18 g, 12.9 mmol, 1.0 eq) in Et.sub.3N (110 mL) was added CuI (49.2 mg, 258 umol, 0.02 eq), PdCl.sub.2(PPh.sub.3).sub.2 (181 mg, 258 umol, 0.02 eq) at 25 C. under N.sub.2 and stirred at 25 C. for 16 hrs. TLC (petroleum ether/ethyl acetate=1/1, R.sub.f=0.3) show that the reaction was complete. Then aq.NH.sub.4Cl (200 mL) was added, extracted with EtOAc (200 mL*3), the combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel eluted with petroleum ether: EtOAc (10:1, 1:1) to give Cpd.4-6 (3.00 g, 48% yield) as yellow oil.

[0185] .sup.1H NMR: ET5008-32-P1b1 400 MHz CDCl.sub.3; 7.41-7.34 (m, 1H), 7.23-7.06 (m, 3H), 4.97-4.87 (m, 1H), 3.97 (s, 2H), 3.53 (t, J=6.8 Hz, 2H), 3.43-3.33 (m, 2H), 2.72 (t, J=8.0 Hz, 2H), 2.64 (J=8.0 Hz, 2H), 1.69-1.59 (m, 4H), 1.55-1.43 (m, 22H).

General Procedure for Preparation of Cpd.4-7

[0186]

TABLE-US-00015 [00072] [00073] Reagent MW. amount mol ratio Other Info. Cpd. 4-6 459.62 3.00 g 6.53 mmol 1.0 eq. Pd/C 200 mg MeOH 20.0 mL Product: (Cpd. 4-7) 463.65 2.50 g 4.91 mmol Yield: 75%

[0187] A mixture of Cpd.4-6 (3.00 g, 6.53 mmol, 1.0 eq) in MeOH (20.0 mL) was added Pd/C (200 mg) and stirred at 25 C. under 50 psi of H.sub.2 for 5 hrs. LC-MS (ET5008-33-P1A4, product: RT=1.04 min) show that the reaction was completed. Then the solution was filtered and concentrated to give Cpd.4-7 (2.50 g, 75% yield) as yellow oil.

[0188] .sup.1H NMR: ET5008-33-P1b1 400 MHz CDCl.sub.3; 7.13 (s, 4H), 4.54 (s, 1H), 3.96 (s, 2H), 3.52 (t, J=6.8 Hz, 2H), 3.18-3.14 (m, 2H), 2.65-2.57 (m, 4H), 1.75-1.54 (m, 10H), 1.53-1.37 (m, 20H).

General Procedure for Preparation of Cpd.4-10

[0189]

TABLE-US-00016 [00074] [00075] Reagent MW. amount mol ratio Other Info. Cpd. 4-7 463.65 1.00 g 2.16 mmol 1.0 eq. HCl/EtOAc 30.0 mL 4N Product: (Cpd. 4-10) 343.89 800 mg 2.33 mmol crude

[0190] A mixture of Cpd.4-7 (1.00 g, 2.16 mmol, 1.0 eq) in HCl/EtOAc (30.0 mL) at 50 C. and stirred at 50 C. for 0.5 h. LC-MS (ET5008-34-P1A4, product: RT=0.698 min) show that the reaction was completed. The mixture was concentrated to give crude Cpd.4-10 (800 mg) as yellow solid.

General Procedure for Preparation of Comp-05

[0191]

TABLE-US-00017 [00076] [00077] Reagent MW. amount mol ratio Other Info. Cpd. 4-10 343.89 800 mg 2.33 mmol 1.0 eq. Cpd. R1 131.13 611 mg 4.66 mmol 2.0 eq. Et.sub.3N 101.19 2.36 g 23.3 mmol 10 eq. EtOH 40.0 mL Product: Comp-05 392.49 370 mg 933 umol Yield: 40%

[0192] A mixture of Cpd.4-10 (800 mg, 2.33 mmol, 1.0 eq) in EtOH (40.0 mL) was added Et.sub.3N (2.36 g, 23.3 mmol, 10.0 eq) and Cpd.R1 (611 mg, 4.66 mmol, 2.0 eq) at 25 C. Then the solution was stirred at 60 C. for 20 hrs. LC-MS (ET5008-35-P1A1, product: RT=0.81 min) show that the reaction was completed. The solution was concentrated. The residue was purified by prep-HPLC (TFA condition) to give Comp-05 (370 mg, 40% yield) as white solid.

HPLC Separation Method:

[0193]

TABLE-US-00018 Column Luna C18 100*30 5u Condition 0.05% HCl-ACN Begin B 30 End B 60 Gradient Time 12 min 100% B Hold Time 4 min Flow Rate 25 mL/min Injection 12

[0194] .sup.1H NMR: ET5008-35-P1b1 400 MHz CDCl.sub.3; 10.46 (br, 1H), 8.35 (s, 1H), 7.74 (s, 1H), 7.12 (s, 4H), 4.12 (s, 2H), 3.59 (t, J=6.0 Hz, 2H 2H), 3.35 (q, J=7.2 Hz, 2H), 2.92 (d, J=5.2 Hz, 3H), 2.65-2.57 (m, 4H), 1.68-1.44 (m, 14H).

For the synthesis of Comp-14 to Comp-34 compounds, general building blocks have been synthezised beforehand:

Building Block 1 (BB-1)

N[(5Z)-6-iodohex-5-en-1-yl]-Nmethylethanediamide

[0195] Step 1:

[0196] PPh3 (140 g) and phthalimide (82.5 g) were suspended in dry THF (500.0 mL) and cooled to 0 C. A solution of 5-hexyn-1-ol (50.0 g) and diisopropyl azodicarboxylate (110 mL) in dry THF (100 mL) was then added dropwise over a period of 45 min. The resulting mixture was stirred at 0 C. for 1 h and then at r.t. over night.

[0197] THF was removed in vacuo as far as possible. The residue was suspended in PE/EtOAc=9:1 (700 mL) and stirred vigourously. The solvent was decanted off from the precipitated OPPh3. During this process, white needles (product) formed in the decanted solvent, which were filtered off and set aside (F1).

[0198] The OPPh3 precipiate was then further washed with PE/EtOAc=9:1 several times. All filtrates were then combined and evaporated in vacuo (F2). The needles from F1 were dissolved in EtOAc (200 mL) and washed with 1N NaOH (275 mL) and brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was filtered through a patch of SiO 2 (eluent CH2Cl2). The solvent was removed in vacuo and the oily residue was left standing in the fridge over weekend, after which white needles had been formed. The mixture was diluted with PE, the product was then filtered off, washed with PE and dried in vacuo to afford F1 as white needles. The mother liquor was combined with F2.

[0199] The yellow oil of F2 was dissolved in EtOAc (400 mL) and washed with 1 N NaOH (3150 mL) and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO 2 (eluent CH2Cl2). The product containing fractions were combined and evaporated. PE was added to the yellow oily residue, after which a precipitate formed. The mixture was cooled to 0 C., the solid was then filtered off and washed with PE to afford F2 as white solid. The mother liquor was evaporated. PE was added to the oily residue after which a precipitate formed. The mixture was left standing in the fridge for 2 h, the precipitate was then filtered off, washed with PE and dried in vacuo to afford F3 as pale yellow solid.

[0200] Step 2:

[0201] 2-(hex-5-yn-1-yl)-2,3-dihydro-1H-isoindole-1,3-dione (46.3 g), AgNO 3 (8.65 g) and NIS (68.8 g) were placed in a 1 L flask. Dry THF (500 mL) was added, the flask was flushed with argon and wrapped with aluminium foil to protect the reaction from light. The mixture was then stirred under an Ar-atmosphere at r.t. for 16 h. Control by LC/MS showed product.

[0202] The reaction mixture was decanted from the formed precipitate, diluted with water (400 mL) and extracted with EtOAc (3200 mL). The combined organic layers were washed with water (100 mL), sat. Na 2SO3 (3100 mL) and brine (100 mL), dried over Na2SO4 and concentrated in vacuo. The residue was recrystallized from EtOH to afford F1 as white solid. The mother liquor was evaporated and again recrystallized from EtOH to afford F2 as yellow solid.

[0203] Step 3:

[0204] 2-Methyl-2-butene (29.4 mL) was added dropwise to a 0 C. cold solution of BH 3*SMe2 (2.00 M in THF, 64.4 mL) and stirred at 0 C. for 1 h and then at r.t. for 1 h. The mixture was then added dropwise to a 0 C. cold suspension of 2-(6-iodohex-5-yn-1-yl)-2,3-dihydro-1H-isoindole-1,3-dione (17.5 g) in THF (200 mL). After addition, the resulting mixture was stirred at r.t. for 1 h. Control by LC/MS showed complete consumption of starting material. The reaction mixture was cooled to 0 C., then HOAc (30.0 mL) was added dropwise, stirred for 30 min at 0 C. and then at r.t. over night. Control by LC/MS showed product.

[0205] THF was removed in vacuo as far as possible. The residue was then slowly poured into a solution of NaOH (15.0 g) in H.sub.2O (200 mL) and extracted with CH2Cl2 (3100 mL). The combined organic layers were washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was used for further transformation as such.

[0206] Step 4:

[0207] 2-[(5Z)-6-iodohex-5-en-1-yl]-2,3-dihydro-1H-isoindole-1,3-dione (17.6 g, crude IK-0353/4) was dissolved in MeOH (150 mL). Hydrazine hydrate (6.00 mL) was added and the resulting mixture was stirred at r.t. for 16 h. Control by LC/MS showed product.

[0208] MeOH was removed in vacuo. The residue was suspended in CH2Cl2 (300 mL). The solid was filtered off and washed with CH2Cl2 (2100 mL). The combined filtrates were then washed with water (2100 mL), dried over Na 2SO4 and concentrated in vacuo to afford the crude product as orange oil which was used for further transformation as such.

[0209] Step 5:

[0210] Ethylchlorformylformiat (10.0 g) was dissolved in THF (50 mL) and cooled to 0 C. Pyridine (7.70 mL) was added dropwise and the mixture stirred at 0 C. for 30 min. Methylamine (2.0 M in THF, 47.6 mL) was then added dropwise. Stirring was continued at 0 C. for 3 h. Control by TLC (PE/EtOAc=1:3) showed product.

[0211] The precipitated salt was filtered off and the filtrate evaporated. The residue was taken up in EtOAc (200 mL), washed with 1N HCl (250 mL), dried over Na 2SO4 and concentrated in vacuo to give the product in sufficient purity as brown oil.

[0212] Step 6:

[0213] (5Z)-6-iodohex-5-en-1-amine (11.15 g) was dissolved in EtOH (200 mL). ethyl (methylcarbamoyl)formate (6.50 g) and NEt3 (8.26 mL) were added and the resulting mixture was stirred at 50 C. for 24 h. Control by LC/MS showed incomplete conversion. Additional (methylcarbamoyl)formate (1.00 g) and NEt3 (4.00 mL) were added and stirring was continued at 50 C. for 24 h. Control by LC/MS showed product.

[0214] EtOH was removed in vacuo. The residue was purified by column chromatography on SiO2 (CH2Cl2->CH2Cl2/MeOH=50:1->CH2Cl2/MeOH=20:1). The product containing fractions were combined and evaporated. EtOAc (30 mL) was added to the partly solid residue, treated with sonication and left standing in the fridge over weekend. The precipitate was then filtered off, washed with little ice cold EtOAc and dried in vacuo.

[0215] Yield: 10.3 g (67%) pale yellow solid.

Building Block 2 (BB-2)

N-[4-(2-iodophenyl)butyl]-Nmethylethanediamide

[0216] Step 1:

[0217] PPh3 (95.5 g), phthalimide (56.1 g) and 3-Buten-1-ol (25.0 g) were suspended in dry THF (250 mL) and cooled to 0 C. Diisopropyl azodicarboxylate (75.1 mL) was then added dropwise over a period of 20 min. The resulting mixture was stirred at 0 C. for 30 min and then at r.t. over night. Control by LC/MS showed product.

[0218] THF was removed in vacuo as far as possible. The oily residue was diluted with PE/EtOAc=9:1 (400 mL) and stirred vigourously until a precipitate occurred. The precipitated OPPh3 was filtered off and washed extensively with PE/EtOAc=9:1. The combined filtrates were filtered through a patch of SiO 2 and then evaporated. The residue was diluted with PE (200 mL), mixed vigourously and placed in an icebath. The precipitated product was then filtered off and washed with PE to afford the product in sufficient purity as pale yellow solid.

[0219] Step 2:

[0220] 2-(but-3-en-1-yl)-2,3-dihydro-1H-isoindole-1,3-dione (22.1 g) was placed in a 1 L flask under and Ar-atmosphere. 9-BBN (0.5 M in THF, 273 mL) was then added dropwise at 0 C. and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. over night. A solution of Na2CO3 (48.4 g) in water (250 mL) was then added and stirring was continued at r.t. for 30 min. Then 2-Iodo-phenylamine (20.0 g) and PdCl2(PPh3)2 (2.80 g) was added and the mixture heated to 50 C. for 4 h. Control by LC/MS showed product.

[0221] The reaction mixture was diluted with EtOAc (200 mL) and the layers separated. The aqueous layer was extracted with EtOAc (300 mL) and the combined organic layers were washed with brine (200 mL) and dried over Na2SO4. The residue was purified by column chromatography on SiO2 (PE/EtOAc=6:4).

[0222] Step 3:

[0223] 2-[4-(2-aminophenyl)butyl]-2,3-dihydro-1H-isoindole-1,3-dione (22.0) was dissolved in acteone (100 mL). Then water (200 mL) and conc. H2SO4 (13.9 mL) were added and the resulting suspension was cooled to 0 C. A solution of NaNO2 (5.23 g) in water (50 mL) was added dropwise and the mixture stirred at 0 C. for 30 min. Then a solution of KI (37.2 g) in water (50 mL) was added dropwise, the reaction mixture warmed to r.t. and stirred for 20 h. Control by LC/MS showed product.

[0224] The reaction mixture was diluted with sat. Na 2503 (200 mL) and extracted with EtOAc (3200 mL). The combined organic layers were washed with brine (150 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=8:2).

[0225] Step 4:

[0226] 2-[4-(2-iodophenyl)butyl]-2,3-dihydro-1H-isoindole-1,3-dione (21.2 g) was suspended in MeOH (300 mL). Hydrazine hydrate (5.10 mL) was added and the resulting mixture was stirred at r.t. for 3d. Control by LC/MS showed product.

[0227] MeOH was removed in vacuo. The residue was suspended in CH2Cl2 (200 mL). The solid was filtered off and washed with CH2Cl2 (100 mL). The combined filtrates were then washed with water (2100 mL). The combined aqueous layers were reextracted with CH2Cl2 (50 mL) and the combined organic layers were then dried over Na2SO4 and concentrated in vacuo to afford the product in sufficient purity as yellow oil.

[0228] Step 5:

[0229] Ethylchlorformylformiat (10.0 g) was dissolved in THF (50 mL) and cooled to 0 C. Pyridine (7.70 mL) was added dropwise and the mixture stirred at 0 C. for 30 min. Methylamine (2.0 M in THF, 47.6 mL) was then added dropwise. Stirring was continued at 0 C. for 3 h. Control by TLC (PE/EtOAc=1:3) showed product.

[0230] The precipitated salt was filtered off and the filtrate evaporated. The residue was taken up in EtOAc (200 mL), washed with 1N HCl (250 mL), dried over Na 2SO4 and concentrated in vacuo to give the product in sufficient purity as brown oil.

[0231] Step 6:

[0232] 4-(2-iodophenyl)butan-1-amine (11.0 g, crude IK-0355710) was dissolved in EtOH (100 mL). Ethyl (methylcarbamoyl) formate (5.76 g) and NEt3 (6.67 mL) were added and the resulting mixture was stirred at 50 C. for 18 h. Control by LC/MS showed product.

[0233] The reaction mixture was cooled to r.t. and EtOH was removed in vacuo. The residue was filtered through a patch of SiO2 (CH2Cl2/MeOH=98:2). Further purification by recrystallization from EtOAc.

[0234] Yield: 7.76 g (54%) beige solid.

Building Block 4 (BB-4)

2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid

[0235] Step 1

[0236] NaH (60% in mineral oil, 771 mg) was suspended in dry THF (20.0 mL). The mixture was cooled to 0 C., then 6-Hepten-1-ol (1.18 mL) was added. Stirring was continued at 0 C. for 30 min, then a solution of bromoacetic acid (1.34 g) in THF (10.0 mL) was added dropwise. After complete addition, the ice bath was removed and stirred for 15 min, then the mixture was heated to 70 C. for 1.5 h. Control by TLC (PE/EtOAc=1:1) showed product.

[0237] The reaction mixture was poured into 1N NaOH (50 mL) and extracted with EtOAc (230 mL). The combined organic layers contained no product and were discarded. The aqueous layer was carefully acidified with conc. HCl and then again extracted with EtOAc (330 mL). The combined organic layers were dried over Na 2SO4 and concentrated in vacuo to afford the product in sufficient purity as colorless oil.

[0238] Step 2

[0239] 1,1-Carbonyldiimidazole (15.6 g) was suspended in THF (200 mL). A solution of 2-(hept-6-en-1-yloxy)acetic acid (15.1 g) in THF (20 mL) was then added dropwise and the resulting mixture was stirred at r.t. for 6 h. THF was then removed in vacuo and MeOH (200 mL) was added to the residue. The mixture was stirred at r.t. for 3d. Control by TLC (PE/EtOAc=9:1) showed product.

[0240] MeOH was removed in vacuo. PE (200 mL) was added to the residue and stirred vigourously for 5 min. The solvent was then decanted off from a thick, oily residue, which was further washed with PE (2100 mL) and then discarded. The combined PE fractions were washed with 1N HCl (100 mL) and 1N NaOH (100 mL), dried over Na2SO4 and concentrated in vacuo to afford the product in sufficient purity as colorless liquid.

[0241] Step 3

[0242] Methyl 2-(hept-6-en-1-yloxy)acetate (2.88 g) was placed in 100 mL flask and cooled to 0 C. under and Aratmosphere. 9-BBN (0.5 M in THF, 38.7 mL) was then added dropwise and the resulting mixture stirred at 0 C. for 30 min and then at r.t. for 2 h. A solution of Na 2CO3 (6.84 g) in water (30.0 mL) was then added and stirring was continued at r.t. for 30 min. Then N[(5Z)-6-iodohex-5-en-1-yl]-N-methylethanediamide (BB-1, 4.00 g) and PdCl2(PPh3)2 (453 mg) were added and the mixture heated to 50 C. for 1.5 h. Control by LC/MS showed product.

[0243] The reaction mixture was cooled to r.t. and the layers were separated. The aqueous layer was extracted with EtOAc (100 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc 1:1).

[0244] Step 4

[0245] Methyl 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetate (400 mg) was suspended in MeOH (20.0 mL). NaOH (3N, 5.00 mL) was added and the resulting mixture was stirred at r.t. for 15 min. Control by LC/MS showed product.

[0246] The reaction mixture was poured into 1N HCl (30 mL). The precipitated product was filtered off, washed with water and dried in vacuo.

[0247] Yield: 869 mg (86%) beige solid.

Building Block 6 (BB-6)

2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]aceticacid

[0248] Step 1:

[0249] NaH (60% in mineral oil, 15.2 g) was suspended in dry THF (250 mL). The mixture was cooled to 0 C., then allylalcohol (11.8 mL) was added. Stirring was continued at 0 C. for 30 min, then a solution of bromoacetic acid (26.3 g) in THF (50.0 mL) was added dropwise. After complete addition, the ice bath was removed and stirred for 15 min, the mixture was then heated to 70 C. for 3 h and stirred at r.t. over night.

[0250] The reaction mixture was poured into water (250 mL) and extracted with EtOAc (2100 mL). The combined organic layers contained no product and were discarded. The aqueous layer was carefully acidified with conc. HCl and then again extracted with EtOAc (3100 mL). The combined organic layers were dried over Na 2SO4 and concentrated in vacuo to afford the product in sufficient purity as pale brown liquid.

[0251] Step 2:

[0252] 1,1-Carbonyldiimidazole (30.7 g) was suspended in THF (200 mL). 2-(prop-2-en-1-yloxy)acetic acid (crude IK-0352/9) was then added dropwise and the resulting mixture was stirred at r.t. for 7 h. THF was then removed in vacuo and MeOH (200 mL) was added to the residue. The mixture was stirred at r.t. over night. Control by TLC (PE/EtOAc=8:2) showed product.

[0253] MeOH was removed in vacuo. PE (200 mL) was added to the residue and stirred vigourously for 5 min. The solvent was then decanted off from a thick, oily residue, which was further washed with PE (2100 mL). Control by TLC showed most of the product remaining in the oily residue, which was thus washed with MTBE (4100 mL). The PE and MTBE layers were combined and washed with 1N HCl (3100 mL) and brine (50 mL), dried over Na2SO4 and concentrated in vacuo to afford the product in sufficient purity as pale yellow liquid.

[0254] Step 3:

[0255] methyl 2-(prop-2-en-1-yloxy)acetate (1.30 g) was placed in a 100 mL flask and cooled to 0 C. under and Aratmosphere. 9-BBN (0.5 M in THF, 25.0 mL) was then added dropwise and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 2 h. A solution of Na 2CO3 (4.41 g) in water (25.0 mL) was then added and stirring was continued at r.t. for 30 min. Then N-[4-(2-iodophenyl)butyl]-N-methylethanediamide (BB-2, 3.00 g) and PdCl 2(PPh3)2 (292 mg) were added and the mixture heated to 50 C. for 4 h and then stirred at r.t. overnight. Control by LC/MS showed incomplete conversion. Additional methyl 2-(prop-2-en-1-yloxy)acetate (650 mg) was placed in a separate flask under an Ar-atmosphere. 9-BBN (0.5 M in THF, 12.5 mL) was added at r.t. and the mixture stirred at r.t. for 2 h. A sat. solution of Na 2CO3 (10 mL) was added and stirring was continued at r.t. for 30 min. The mixture was then added to the above reaction mixture. After adding fresh PdCl2(PPh3)2 (200 mg), the mixture was stirred at 50 C. for 2 h. Control by LC/MS showed product.

[0256] The reaction mixture was cooled to r.t. and the layers were separated. The aqueous layer was extracted with EtOAc (100 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc 3:7).

[0257] Step 4:

[0258] methyl 2-[3-(2-{4-[(methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetate (2.04 g) was dissolved in THF (30 mL). NaOH (3N, 30 mL) and MeOH (20 mL) were added and the resulting mixture was stirred at r.t. for 5 min. Control by LC/MS showed product.

[0259] The reaction mixture was acidified with 6N HCl and extracted with EtOAc (340 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by a short column on SiO2 (CH2Cl2/MeOH=9:1).

[0260] Yield: 1.56 g (80%) beige solid.

Building Block 8 (BB-8)

N[(5Z)-13-hydroxytridec-5-en-1-yl]-Nmethylethanediamide

[0261] Step 1:

[0262] 6-Hepten-1-ol (3.00 g) and imidazole (3.57 g) were dissolved in DMF (20.0 mL). TIPSCl (6.18 mL) was added and the resulting mixture was stirred at 60 C. for 6 h. Control by TLC (PE/EtOAc=8:2) showed almost complete conversion.

[0263] The reaction mixture was diluted with water (100 mL) and extracted with MTBE (340 mL). The combined organic layers were washed with 1N HCl (250 mL) and brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=95:5).

[0264] Step 2:

[0265] (hept-6-en-1-yloxy)tris(propan-2-yl)silane (1.57 g) was placed in 100 mL flask and cooled to 0 C. under and Aratmosphere. 9-BBN (0.5 M in THF, 14.5 mL) was then added dropwise and the resulting mixture stirred at 0 C. for 30 min and then at r.t. for 2 h. A solution of Na 2CO3 (2.56 g) in water (15.00 mL) was then added and stirring was continued at r.t. for 30 min. then N[(5Z)-6-iodohex-5-en-1-yl]-N-methylethanediamide (BB-1, 1.50 g) and PdCl2(PPh3)2 (170 mg) were added and the mixture heated to 50 C. for 2 h. Control by LC/MS showed product.

[0266] The reaction mixture was cooled to r.t. and the layers were separated. The aqueous layer was extracted with EtOAc (250 mL). The combined organic layers were washed with brine (30 mL), dried over Na 2SO4 and concentrated in vacuo. The residue was filtered through a patch of SiO 2 (PE/EtOAc=4:6). The so obtained crude product was used for further transformation as such.

[0267] Step 3:

[0268] (hept-6-en-1-yloxy)tris(propan-2-yl)silane (2.20 g, crude 1K-0357/16) was dissolved in THF (50 mL) and cooled to 0 C. TBAF*3H.sub.2O (2.29 g) was added and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 6 h. Control by TLC (PE/EtOAc=1:1) and LC/MS showed complete conversion.

[0269] The reaction mixture was poured into water (100 mL) and extracted with EtOAc (340 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was passed through a short column on SiO2 (PE/EtOAc=1:1->EtOAc).

[0270] Yield: 1.11 g (77%) beige solid.

Building Block 9 (BB-9)

N-{4-[2-(3-hydroxypropyl)phenyl]butyl}-Nmethylethanediamide

[0271] Step 1:

[0272] 2-Propen-1-ol (3.00 g) and imidazole (7.03 g) were dissolved in DMF (20.0 mL). TIPSCl (14.4 mL) was added and the resulting mixture was stirred at 60 C. for 6 h. Control by TLC (PE/EtOAc=8:2) showed almost complete conversion. The reaction mixture was diluted with water (100 mL) and extracted with MTBE (340 mL). The combined organic layers were washed with 1N HCl (250 mL) and brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=95:5).

[0273] Step 2:

[0274] (prop-2-en-1-yloxy)tris(propan-2-yl)silane (1.33 g) was placed in a 100 mL flask and cooled to 0 C. under and Ar-atmosphere. 9-BBN (0.5 M in THF, 14.2 mL) was then added dropwise and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 2 h.

[0275] A solution of Na 2CO3 (2.21 g) in water (15.0 mL) was then added and stirring was continued at r.t. for 30 min. Then N-[4-(2-iodophenyl)butyl]-N-methylethanediamide (1.50 g) and PdCl 2(PPh3)2 (146 mg) were added and the mixture heated to 50 C. for 3 h. Control by LC/MS showed product. The reaction mixture was cooled to r.t. and the layers were separated. The aqueous layer was extracted with EtOAc (100 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was passed through a short column of SiO 2 (PE/EtOAc 1:1). The still crude product was then used for further transformation as such.

[0276] Step 3:

[0277] N-methyl-N-{4-[2-(3-{[tris(propan-2-yl)silyl]oxy}propyl)phenyl]butyl}ethanediamide (1.87 g, crude IK-0357/17) was dissolved in THF (50 mL) and cooled to 0 C. TBAF*3H.sub.2O (1.97 g) was added and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 16 h. Control by LC/MS showed complete conversion. The reaction mixture was poured into water (100 mL) and extracted with EtOAc (340 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was passed through a short column on SiO2 (PE/EtOAc=1:1->EtOAc).

[0278] Yield: 911 mg (75%) beige solid.

Building Block 11 (BB-11)

N[(5Z)-13-(2-aminoethoxy)tridec-5-en-1-yl]-Nmethylethanediamide

[0279] Step 1:

[0280] NaH (60% in mineral oil, 7.71 g) was suspended in dry THF (200 mL). The mixture was cooled to 0 C., then 6-Hepten-1-ol (11.8 mL) was added. Stirring was continued at 0 C. for 30 min, then a solution of bromoacetic acid (13.4 g) in THF (100 mL) was added dropwise. After complete addition, the ice bath was removed and stirred for 15 min, then the mixture was heated to 70 C. for 3 h. Control by TLC (PE/EtOAc=1:1) showed product.

[0281] The reaction mixture was poured into 1N NaOH (300 mL) and extracted with EtOAc (2100 mL). The combined organic layers contained no product and were discarded. The aqueous layer was carefully acidified with conc. HCl and then again extracted with EtOAc (3100 mL). The combined organic layers were dried over Na 2SO4 and concentrated in vacuo to afford the product in sufficient purity as pale brown oil.

[0282] Step 2:

[0283] 1,1-Carbonyldiimidazole (15.6 g) was suspended in THF (200 mL). A solution of 2-(hept-6-en-1-yloxy)acetic acid (15.1 g) in THF (20 mL) was then added dropwise and the resulting mixture was stirred at r.t. for 6 h. THF was then removed in vacuo and MeOH (200 mL) was added to the residue. The mixture was stirred at r.t. for 3d. Control by TLC (PE/EtOAc=9:1) showed product.

[0284] MeOH was removed in vacuo. PE (200 mL) was added to the residue and stirred vigourously for 5 min. The solvent was then decanted off from a thick, oily residue, which was further washed with PE (2100 mL) and then discarded. The combined PE fractions were washed with 1N HCl (100 mL) and 1N NaOH (100 mL), dried over Na2SO4 and concentrated in vacuo to afford the product in sufficient purity as colorless liquid.

[0285] Step 3:

[0286] methyl 2-(hept-6-en-1-yloxy)acetate (5.00 g) was dissolved in CH 2Cl2 (100 mL) and cooled to 0 C. DIBALH (1.00 M in CH2Cl2, 61.7 mL) was added dropwise, and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. over night. Control by TLC (PE/EtOAc=8:2) showed complete conversion.

[0287] The reaction mixture was cooled to 0 C. and carefully quenched with sat. aqueous Na 2SO4. The mixture was then diluted with CH2Cl2 (100 mL), stirred vigourously for 20 min and then filtered through celite. The filtercake was washed with CH2Cl2 several times. The combined filtrates were concentrated in vacuo to afford the product in sufficient purity as colorless liquid.

[0288] Step 4:

[0289] PPh3 (7.17 g), phthalimide (4.21 g) and 2-(hept-6-en-1-yloxy)ethan-1-ol (4.12 g) were suspended in dry THF (100 mL) and cooled to 0 C. Diisopropyl azodicarboxylate (5.79 mL) was then added dropwise over a period of 20 min. The resulting mixture was stirred at 0 C. for 30 min and then at r.t. over night.

[0290] THF was removed in vacuo as far as possible. The oily residue was diluted with PE/EtOAc=9:1 (200 mL) and stirred vigourously until a precipitate occurred. The precipitated OPPh3 was filtered off and washed extensively with PE/EtOAc=9:1. The combined filtrates were filtered through a patch of SiO2 (eluent PE/EtOAc=9:1) and evaporated. The residue was purified by column chromatography on SiO2 (PE/EtOAc=8:2).

[0291] Step 5:

[0292] 2-[2-(hept-6-en-1-yloxy)ethyl]-2,3-dihydro-1H-isoindole-1,3-dione (2.22 g) was placed in 100 mL flask and cooled to 0 C. under and Ar-atmosphere. 9-BBN (0.5 M in THF, 19.3 mL) was then added dropwise and the resulting mixture stirred at 0 C. for 30 min and then at r.t. for 2 h. A solution of Na2CO3 (3.42 g) in water (20.0 mL) was then added and stirring was continued at r.t. for 30 min. then N [(5Z)-6-iodohex-5-en-1-yl]-N-methylethanediamide (BB-1, 2.00 g) and PdCl 2(PPh3)2 (226 mg) were added and the mixture heated to 50 C. for 1.5 h. Control by LC/MS showed product.

[0293] The reaction mixture was cooled to r.t. and the layers were separated. The aqueous layer was extracted with EtOAc (230 mL). The combined organic layers were washed with brine (30 mL), dried over Na 2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=4:6).

[0294] Step 6:

[0295] N[(5Z)-13-[2-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)ethoxy]tridec-5-en-1-yl]-N-methylethanediamide (2.36 g) was suspended in MeOH (100 mL). Hydrazine hydrate (486 L) was added and the resulting mixture was stirred at r.t. for 16 h. Control by LC/MS showed product.

[0296] MeOH was removed in vacuo. The residue was suspended in CH 2Cl2/7N NH3 in MeOH=9:1 (100 mL) and filtered through as patch of SiO2 and further eluated with CH2Cl2/7N NH3 in MeOH=9:1 (300 mL). The filtrate was concentrated in vacuo to afford 1.68 g of the crude product. 60 mg were subjected to purification by preparative TLC (CH2Cl2/7N NH3 in MeOH=9:1). The rest of the crude material was used for further transformations as such.

[0297] Yield: 41 mg (2%) pale yellow solid (purified).

Compound 14 (Comp-14)

N-methyl-N[(5Z)-13-[(1H-1,2,3,4-tetrazol-5-yl)methoxy]tridec-5-en-1-yl] ethanediamide Step 1

[0298] NaH (60% in mineral oil, 7.71 g) was suspended in dry THF (200 mL). The mixture was cooled to 0 C., then 6-Hepten-1-ol (11.8 mL) was added. Stirring was continued at 0 C. for 30 min, then a solution of bromoacetic acid (13.4 g) in THF (100 mL) was added dropwise. After complete addition, the ice bath was removed and stirred for 15 min, then the mixture was heated to 70 C. for 3 h. Control by TLC (PE/EtOAc=1:1) showed product. The reaction mixture was poured into 1N NaOH (300 mL) and extracted with EtOAc (2100 mL). The combined organic layers contained no product and were discarded. The aqueous layer was carefully acidified with conc. HCl and then again extracted with EtOAc (3100 mL). The combined organic layers were dried over Na 2SO4 and concentrated in vacuo to afford the product in sufficient purity as pale brown oil.

[0299] Yield: 14.1 g (93%) pale brown oil

[0300] Step 2

[0301] A mixture of 2-(hept-6-en-1-yloxy)acetic acid (3.00 g) and SOCl 2 (15.00 mL) was heated to 70 C. for 1 h. Excess SOCl2 was then removed in vacuo and the residue was taken up in dichloroethane (15.0 ml). Then ammonia was slowly bubbled through the solution for 5 min. The reaction mixture diluted with water (50 mL) and extracted with CH2Cl2 (330 mL). The combined organic layers were washed with sat. NaHCO.sub.3(30 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo to afford the product in sufficient purity as white solid. m=2,16 g (y=62%). Analog in TLC to IK-0367/1

[0302] Step 3

[0303] 2-(hept-6-en-1-yloxy)acetamide (2,61 g) was dissolved in CH2Cl2 (50 mL). NEt3 (6.35 mL) was added and the mixture was cooled to 0 C. A solution of POCl3 (1.54 mL) in CH2Cl2 (4 mL) was slowly added. Stirring was then continued at 0 C. for 15 min. Control by TLC (PE/EtOAc=8:2) showed product.

[0304] sat. NaHCO.sub.3(5.00 mL) was added at 0 C. and stirred for 30 min at that temperature. The mixture was allowed to come to r.t., diluted with water (15.0 mL) and extracted with CH2Cl2 (320 mL). The combined organic layers were washed with sat. NaHCO.sub.3(10.0 mL) and brine (10.0 mL), dried over Na2SO4 and then filtered through a pad of SiO2 (eluent CH2Cl2). The product was obtained after evaporation in sufficient purity as colorless oil. m=2,08 g, y=89%.

[0305] Step 4

[0306] 2-(hept-6-en-1-yloxy)acetonitrile was placed in a 10 mL flask and cooled to 0 C. under and Ar-atmosphere. 9-BBN (0.5 M in THF, 1.63 mL) was then added dropwise and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 2 h. A solution of Na2CO3 (288 mg) in degazed water (1 mL) was then added and stirring was continued at r.t. for 30 min. Then N[(5Z)-6-iodohex-5-en-1-yl]-N-methylethanediamide (168 mg) and PdCl2(PPh3)2 (19 mg) were added and the mixture heated to 50 C. overnight. Water was added and mixture was extracted with DCM. Organic layer was dried over MgSO4, filtered and solvent evaporated. Mixture was purified by preparative TLC (DCM/MeOH 95/5). m=70 mg, y=38%.

[0307] Step 5

[0308] N[(5Z)-13-(cyanomethoxy)tridec-5-en-1-yl]-N-methylethanediamide, natrium azide and triethylamine hydrochloride were dissolved in THF and the reaction mixture stirred at 70 C. overnight.

[0309] Water and ethyl acetate were added. The mixture was acidified with HCl3N. The aqueous layer (acid pH) is then extracted with ethyl acetate (3), and the combined organic layer washed with brine. The organic layer was dried over MgSO4, filtered and solvent removed under vacuo. m=82 mg. Product was purified by preparative TLC (DCM/MeOH 95/5)

[0310] Yield: 8 mg (10%), as white powder.

Compound 15 (Comp-15)

N-(4-{2-[3-(carbamoylmethoxy)propyl]phenyl}butyl)-N-methylethanediamide

[0311] 250 mg (0.72 ol) 2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic acid (BB-6) and 164.1 mg (0.86 mmol) EDCI were dissolved in 20 ml DCM. 36.5 mg (2.14 mmol,

[0312] 5.35 ml) ammonia (0.4 M in THF) were added and the mixture was stirred at rt over the weekend.

[0313] The mixture was poured into 50 ml water and extracted with DCM (350 ml). The combined organic layers were dried over Na2SO4 and concentrated.

[0314] Yield: 50 mg (20%), white solid.

Compound 16 (Comp-16)

N-Methyl-N[(5Z)-13-[(5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy]tridec-5-en-1-yl]ethanediamide

[0315] Step 1:

[0316] NaH (60% in mineral oil, 7.71 g) was suspended in dry THF (200 mL). The mixture was cooled to 0 C., then 6-Hepten-1-ol (11.8 mL) was added. Stirring was continued at 0 C. for 30 min, then a solution of bromoacetic acid (13.4 g) in THF (100 mL) was added dropwise. After complete addition, the ice bath was removed and stirred for 15 min, then the mixture was heated to 70 C. for 3 h. Control by TLC (PE/EtOAc=1:1) showed product. The reaction mixture was poured into 1N NaOH (300 mL) and extracted with EtOAc (2100 mL). The combined organic layers contained no product and were discarded. The aqueous layer was carefully acidified with conc. HCl and then again extracted with EtOAc (3100 mL). The combined organic layers were dried over Na 2SO4 and concentrated in vacuo to afford the product in sufficient purity as pale brown oil.

[0317] Step 2:

[0318] 1,1-Carbonyldiimidazole (15.6 g) was suspended in THF (200 mL). A solution of 2-(hept-6-en-1-yloxy)acetic acid (15.1 g) in THF (20 mL) was then added dropwise and the resulting mixture was stirred at r.t. for 6 h. THF was then removed in vacuo and MeOH (200 mL) was added to the residue. The mixture was stirred at r.t. for 3d. Control by TLC (PE/EtOAc=9:1) showed product. MeOH was removed in vacuo. PE (200 mL) was added to the residue and stirred vigourously for 5 min. The solvent was then decanted off from a thick, oily residue, which was further washed with PE (2100 mL) and then discarded. The combined PE fractions were washed with 1N HCl (100 mL) and 1N NaOH (100 mL), dried over Na2SO4 and concentrated in vacuo to afford the product in sufficient purity as colorless liquid.

[0319] Step 3:

[0320] 500 mg (2.68 mmol) and 1.34 g (26.9 mmol, 1.30 ml) hydrazine hydrate were dissolved in 5 ml EtOH and stirred at 70 C. for 4.5 h (->clear solution). [0321] LC/MS: GH-0513/1-1 [0322] TLC (EA/PE 1:1): Complete consumption of starting material

[0323] The mixture was evaporated to dryness

[0324] Step 4:

[0325] 500 mg (2.68 mmol) 2-(Hept-6-en-1-yloxy)acetohydrazide (GH-0513/1) were dissolved in 3 ml AcOH. 653.3 mg (8.05 mmol) potassium cyanate dissolved in 3 ml water were added and the mixture was stirred at rt for 1.5 h (a yellow solution). The mixture was evaporated to dryness.

[0326] Step 5:

[0327] The oily residue was dissolved in 10 ml 2M NaOH and heated to reflux for 2 h.->LC/MS: GH-0515/1-2: Complete consumption of intermediate 1. The mixture was acidified using conc. HCl and extracted with EA (320 ml). The combined organic layers were dried over Na2SO4 and concentrated to dryness. The crude solid was recrystallized from ACN.

[0328] Step 6:

[0329] Under Argon atmosphere 92.0 mg (0.44 mmol) 3-[(Hept-6-en-1-yloxy)methyl]-4,5-dihydro-1H-1,2,4-triazol-5-one (GH-0515/1) dissolved in 2 ml anhydrous THF were added to a solution of 88.5 mg (0.73 mmol, 1.45 ml) 9BBN (0.5M in THF) and the mixture was stirred at rt over night. A solution of 153.8 mg (1.45 mmol) Na2CO3 in 1 ml water were added and stirring at rt was continued for 15 min. Then 90.0 mg (0.29 mmol) N[(5Z)-6-iodohex-5-en-1-yl]-methylethanediamide (IK-0356/2) dissolved in 2 ml THF and 10.2 mg (14.5 mol) PdCl 2(PPh3)2 were added and the mixture was heated to 50 C. for 4 h ( yellow biphasic mixture). [0330] LC/MS: GH-0516/1-1: Product was detected

[0331] The organic layer was separated via pipette and evaporated to dryness. The crude product was purified via flash column chromatography on silica gel (DCM/MeOH 20:1 9:1, Rf of possible product: 0.62). Recrystallization from CAN.

[0332] Yield: 51 mg (0.13 mmol, 45%).

Compound 17 (Comp-17)

N-methyl-N[(5Z)-13-[(phenylcarbamoyl)methoxy]tridec-5-en-1-yl]ethanediamide

[0333] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50.0 mg, 140.3 mol), Aniline (26 l, 280.5 mol), HBTU (53.4 mg, 140.3 mol) and DMAP (1.7 mg, 14.0 mol) were placed in a G16 vial. DMF (2.00 ml) and NEt 3 (78.0 l, 561.1 mol) were added and the resulting mixture was stirred at r.t. for 16 h. Control by LC/MS showed product.

[0334] The reaction mixture was diluted with water (20 ml) and was extracted with Et 20 (320 ml). The combined organic layers were washed with sat. NaHCO.sub.3(20 ml) and brine (10 ml), dried over Na2SO4 and concentrated in vacuo. The product was lyophilized.

[0335] Yield: 52 mg (87%), white solid.

Compound 18 (Comp-18)

N-methyl-N[(5Z)-13-{[(oxan-4-yl)carbamoyl]methoxy}tridec-5-en-1-yl] ethanediamide

[0336] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50.0 mg, 140.3 mol), 4-Aminotetrahydropyran (29 l, 280.5 mol), HBTU (53.4 mg, 140.3 mol) and DMAP (1.7 mg, 14.0 mol) were placed in a G16 vial. DMF (2.00 ml) and NEt3 (78.0 l, 561.1 mol) were added and the resulting mixture was stirred at r.t. for 16 h. Control by LC/MS showed product. The reaction mixture was diluted with water (20 ml) and was extracted with Et2O (320 ml). The combined organic layers were washed with sat. NaHCO.sub.3(20 ml) and brine (10 ml), dried over Na2SO4 and concentrated in vacuo. The product was lyophilized.

[0337] Yield: m=58 mg (94%) white solid.

Compound 19 (Comp-19)

N-methyl-N[(5Z)-13-{[(1,3-oxazol-2-yl)carbamoyl]methoxy}tridec-5-en-1-yl]ethanediamide

[0338] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50.0 mg, 140.3 mol), 1,3-Oxazol-2-amine (24 mg, 280.5 mol), HBTU (53.4 mg, 140.3 mol) and DMAP (1.7 mg, 14.0 mol) were placed in a G16 vial. DMF (2.00 ml) and NEt 3 (78.0 l, 561.1 mol) were added and the resulting mixture was stirred at r.t. for 16 h. Control by LC/MS showed product.

[0339] The reaction mixture was diluted with water (20 ml) and was extracted with Et 20 (320 ml). The combined organic layers were washed with sat. NaHCO.sub.3(20 ml) and brine (10 ml), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (DCM/MeOH=95:5).

[0340] Yield: 11 mg (19%), white solid.

Compound 20 (Comp-20)

N[(5Z)-13-{[(4-methoxyphenyl)carbamoyl]methoxy}tridec-5-en-1-yl]-Nmethylethanediamide

[0341] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50.0 mg, 140.3 mol), p-Anisidine (35 mg, 280.5 mol), HBTU (53.4 mg, 140.3 mol) and DMAP (1.7 mg, 14.0 mol) were placed in a G16 vial. DMF (2.00 ml) and NEt3 (78.0 l, 561.1 mol) were added and the resulting mixture was stirred at r.t. for 16 h. Control by LC/MS showed product.

[0342] The reaction mixture was diluted with water (20 ml) and was extracted with Et 20 (320 ml). The combined organic layers were washed with sat. NaHCO.sub.3(20 ml) and brine (10 ml), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (DCM/MeOH=98:2).

[0343] Yield: 18 mg (28%), beige solid.

Compound 21 (Comp-21)

N-Methyl-N-[4-(2-{3-[(phenylcarbamoyl)methoxy]propyl}phenyl)butyl] ethanediamide

[0344] 40 mg (0.11 mmol) 2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic acid (BB-6), 26.3 mg (0.14 mmol) EDCI and 11.7 mg (0.13 mmol, 11.5 l) aniline were dis.Math.sol.Math.ved in 3 ml DCM and stirred at rt of the weekend (clear solution).

[0345] The mixture was evaporated to dryness and purified via pTLC (1 mm, DCM/MeOH 20:1, Rf of possible product: 0.54).

[0346] Yield: 24 mg (51%), white solid.

Compound 22 (Comp-22)

N-Methyl-N-[4-(2-{3-[2-oxo-2-(pyrrolidin-1-yl)ethoxy]propyl}phenyl)butyl] ethanediamide

[0347] 50 mg (0.14 mmol) 2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic acid (BB-6), 32.8 mg (0.17 mmol) EDCI and 20.3 mg (0.29 mmol, 23.4 l) Pyrrolidine were dissolved in 3 ml DCM and stirred at rt for 1.5 h (clear solution).

[0348] The mixture was evaporated to dryness and purified via pTLC (1 mm, DCM/MeOH 10:1, Rf of possible product: 0.46).

[0349] Yield: 20 mg (35%), white solid.

Compound 23 (Comp-23)

N-Methyl-N-[4-(2-{3-[2-(morpholin-4-yl)-2-oxoethoxy]propyl}phenyl)butyl] ethanediamide

[0350] 50 mg (0.14 mmol) 2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic acid (BB-6, IK-0358/6), 32.8 mg (0.17 mmol) EDCI and 24.9 mg (0.29 mmol, 24.9 l) morpholine were dissolved in 3 ml DCM and stirred at rt over the weekend (clear solution).

[0351] The mixture was evaporated to dryness and purified via pTLC (1 mm, DCM/MeOH 10:1, Rf of possible product: 0.48).

[0352] Yield: 34 mg (58%), white solid.

Compound 24 (Comp-24)

N[(5Z)-13-{[(benzenesulfonyl)carbamoyl]methoxy}tridec-5-en-1-yl]-Nmethylethanediamide

[0353] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50.0 mg) was dissolved in THF (2.00 mL). 1,1-Carbonyldiimidazole (29.6 mg) was added and the resulting mixture was stirred at r.t. for 2 h. Then DBU (52.9 L) and benzenesulfonamide (33.1 mg) were added and stirring was continued at r.t. for 25 h. Control by LC/MS showed OMT-121.

[0354] The reaction mixture was diluted with water (30 mL) and extracted with CH 2Cl2 (320 mL). The combined organic layers were washed with 1N HCl (20 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0355] Yield: 44 mg (63%), white solid.

Compound 25 (Comp-25)

4-{2-[3-(2-{4-[(methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetamido}benzoic acid

[0356] Step 1:

[0357] 50 mg (0.14 mmol) 2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic acid (BB-6, IK-0358/6), 32.8 mg (0.17 mmol) EDCI and 43.1 mg (0.29 mmol) Methyl 4-aminobenzoate were dissolved in 3 ml DCM and stirred at rt for 1.5 h (clear solution).

[0358] The mixture was evaporated to dryness and purified via pTLC (1 mm, EA/PE 4:1, R f of possible product: 0.31)

[0359] Step 2:

[0360] To a solution of 48 mg (0.10 mmol)N-Methyl-N-{4-[2-(3-{[(pyridin-2-yl)carbamoyl]methoxy}propyl)phenyl]butyl}ethanediamide (GH-0498/1) in 2 ml THF 16.7 mg (0.40 mmol) LiOH monohydrate dissolved in 0.5 ml water were added and the mixture was stirred at rt over night (a biphasic mixture).

[0361] The mixture was poured into 1 N HCl solution (10 ml) and extracted with DCM (320 ml). The combined organic layers were dried over Na2SO4 and concentrated to dryness. The crude product was purified via pTLC (DCM/MeOH/FA 100:10:1, Rf of possible product: 0.43).

[0362] Yield: 10 mg (21%), white solid.

Compound 26 (Comp-26)

N[(5Z)-13-[2-(4-hydroxy-2-oxo-2,5-dihydrofuran-3-yl)-2-oxoethoxy]tridec-5-en-1-yl]-Nmethylethanediamide

[0363] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50 mg), DCC (34.7 mg), DMAP (22.3 mg) and 2,4(3H,5H)-Furandione (15.4 mg) were placed in a G16 vial. CH2Cl2 (3.00 mL) was added and the resulting mixture was stirred at r.t. for 18 h. Control by LC/MS showed product.

[0364] The reaction mixture was diluted with water (30 mL) and extracted with CH 2Cl2 (320 mL). The combined organic layers were washed with brine (10 mL), dried over Na 2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=9:1).

[0365] Yield: 20 mg (33%), beige solid.

Compound 27 (Comp-27)

N-[4-(2-(3-[2-(hydroxymethyl) phenoxyl]propyl)phenyl)butyl]-N-methylethanediamide

[0366] Step 1:

[0367] Salicylaldehyde (2.00 g) and imidazole (2.79 g) were dissolved in DMF (20.0 mL). TIPSCl (5.96 mL) was added and the resulting mixture was stirred at 60 C. for 2d. Control by TLC (PE/EtOAc=95:5) and LC/MS showed incomplete conversion. Additional TIPSCl (2.00 mL) was added and stirring was continued at 60 C. for 3d. Control by TLC (PE/EtOAc=95:5) and LC/MS showed almost complete conversion. The reaction mixture was diluted with water (100 mL) and extracted with MTBE (340 mL). The combined organic layers were washed with 1N NaOH (30 mL) and brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=95:5).

[0368] Yield: 3.54 g (78%) pale yellow liquid

[0369] Step 2:

[0370] 2-{[tris(propan-2-yl)silyl]oxy}benzaldehyde (3.54 g) was dissolved in EtOH (30.0 mL) and cooled to 0 C. NaBH4 (481 mg) was added and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 18 h. Control by TLC (PE/EtOAc=8:2) and LC/MS showed product. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (340 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=8:2).

[0371] Yield: 2.73 g (77%) yellow oil

[0372] Step 3:

[0373] (2-{[tris(propan-2-yl)silyl]oxy}phenyl)methanol (400 mg) was dissolved in dry THF (15 mL). NaH (60% in mineral oil, 85.6 mg) was added and the mixture stirred at r.t. for 15 min. Allylbromide (309 L) was then added and the resulting mixture was stirred at r.t. over night. Control by LC/MS showed product. The reaction mixture was diluted with water (50 mL) and extracted with CH2Cl2 (330 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The so obtained crude product was used for further transformation as such.

[0374] Yield: 480 mg (crude) yellow oil

[0375] Step 4:

[0376] {2-[(prop-2-en-1-yloxy)methyl]phenoxy}tris(propan-2-yl)silane (178 mg) was placed in a 10 mL flask and cooled to 0 C. under and Ar-atmosphere. 9-BBN (0.5 M in THF, 1.38 mL) was then added dropwise and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 2 h. A solution of Na2CO3 (147 mg) in water (1.50 mL) was then added and stirring was continued at r.t. for 30 min. Then N-[4-(2-iodophenyl)butyl]-N-methylethanediamide (BB-2, 100 mg) and PdCl2(PPh3)2 (9.7 mg) were added and the mixture heated to 50 C. for 3 h. Control by LC/MS showed product. The reaction mixture was cooled to r.t., diluted with water (20 mL) and the layers were separated. The aqueous layer was extracted with EtOAc (15 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was passed through a short column of SiO2 (PE/EtOAc 1:1). The still crude product was then used for further transformation as such.

[0377] Yield: 235 mg (crude) yellow oil.

[0378] Step 5:

[0379] N-methyl-N-[4-(2-{3-[(2-{[tris(propan-2-yl)silyl]oxy}phenyl)methoxy]propyl}phenyl)butyl]ethanediamide (154 mg, crude IK-0357/19) was dissolved in THF (5.00 mL). TBAF*3H 2O (87.0 mg) was added and the resulting mixture was stirred at r.t. for 30 min. Control by LC/MS showed complete conversion. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (310 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0380] Yield: 75 mg (66%) white solid.

Compound 28 (Comp-28)

N[(5Z)-13-(2-benzenesulfonamidoethoxy)tridec-5-en-1-yl]-Nmethylethanediamide

[0381] N[(5Z)-13-(2-aminoethoxy)tridec-5-en-1-yl]-Nmethylethanediamide (BB-11, 50.0 mg, crude IK-0355/8) was suspended in CH2Cl2 (3.00 mL). Benzenesulfonyl chloride (37.5 L) and NEt 3 (61.1 L) were added and the reaction mixture stirred at r.t. for 2 h. Control by LC/MS showed product. Control by LC/MS showed product. Control by LC/MS showed product.

[0382] The reaction mixture was diluted with water (20 mL) and sat. NaHCO 3 (20 mL) and then extracted with CH 2Cl2 (320 mL). The combined organic layers were washed with 1 N HCl (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0383] Yield: 42 mg (60%), white solid.

Compound 29 (Comp-29)

N[(5Z)-13-(2-hydroxyphenoxy)tridec-5-en-1-yl]-Nmethylethanediamide

[0384] Step 1:

[0385] N[(5Z)-13-hydroxytridec-5-en-1-yl]-N-methylethanediamide (BB-8, 400 mg) was suspended in CH 2Cl2 (20 mL). PPh 3 (598 mg) and CBr4 (756 mg) were added and the resulting mixture was stirred at r.t. for 1.5 h. Control by TLC (PE/EtOAc=1:1) and LC/MS showed complete conversion.

[0386] The reaction mixture was concentrated in vacuo and the residue was purified by preparative TLC (PE/EtOAc=1:1).

[0387] Step 2:

[0388] N[(5Z)-13-bromotridec-5-en-1-yl]-N-methylethanediamide (50 mg), pyrocatechol (76.2 mg) and K2CO3 (57.4 mg) were placed in a G16 vial. DMF (3.00 mL) was added and the resulting mixture was stirred at 60 C. for 2.5 h. Control by LC/MS showed product.

[0389] The reaction mixture was diluted with water (40 mL) and extracted with MTBE (320 mL). The combined organic layers were washed with water (20 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0390] Yield: 43 mg (80%), white solid.

Compound 30 (Comp-30)

N-Methyl-N-[4-(2-{3-[2-(morpholin-4-yl)-2-oxoethoxy]propyl}phenyl)butyl] ethanediamide

[0391] 50 mg (0.14 mmol) 2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic acid (BB-8, IK-0358/6), 32.8 mg (0.17 mmol) EDCI and 24.9 mg (0.29 mmol, 24.9 l) morpholine were dissolved in 3 ml DCM and stirred at rt over the weekend (clear solution).

[0392] The mixture was evaporated to dryness and purified via pTLC (1 mm, DCM/MeOH 10:1, Rf of possible product: 0.48).

[0393] Yield: 34 mg (58%), white solid.

Compound 31 (Comp-31)

N-(4-{2-[3-(3-hydroxyphenoxy)propyl]phenyl}butyl)-Nmethylethanediamide

[0394] Step 1:

[0395] N-{4-[2-(3-hydroxypropyl)phenyl]butyl}-N-methylethanediamide (BB-9, 400 mg) was suspended in CH2Cl2 (20 mL). PPh3 (610 mg) and CBr4 (771 mg) were added and the resulting mixture was stirred at r.t. for 30 min. Control by TLC (PE/EtOAc=1:1) and LC/MS showed complete conversion. The reaction mixture was concentrated in vacuo and the residue was purified by preparative TLC (PE/EtOAc=1:1).

[0396] Step 2:

[0397] N-{4-[2-(3-bromopropyl)phenyl]butyl}-N-methylethanediamide (50 mg), 1,3-Benzenediol (77.5 mg) and K2CO3 (58.4 mg) were placed in a G16 vial. DMF (3.00 mL) was added and the resulting mixture was stirred at 60 C. for 1 h. Control by LC/MS showed product.

[0398] The reaction mixture was diluted with water (40 mL) and extracted with MTBE (320 mL). The combined organic layers were washed with water (20 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0399] Yield: 40 mg (74%), white solid.

Compound 32 (Comp-32)

N[(5Z)-13-(4-hydroxyphenoxy)tridec-5-en-1-yl]-Nmethylethanediamide

[0400] Step 1:

[0401] N[(5Z)-13-hydroxytridec-5-en-1-yl]-N-methylethanediamide (BB-8, 400 mg) was suspended in CH 2Cl2 (20 mL). PPh 3 (598 mg) and CBr4 (756 mg) were added and the resulting mixture was stirred at r.t. for 1.5 h. Control by TLC (PE/EtOAc=1:1) and LC/MS showed complete conversion.

[0402] The reaction mixture was concentrated in vacuo and the residue was purified by preparative TLC (PE/EtOAc=1:1).

[0403] Step 2:

[0404] N[(5Z)-13-bromotridec-5-en-1-yl]-N-methylethanediamide (50 mg), hydroquinone (76.2 mg) and

[0405] K2CO3 (57.4 mg) were placed in a G16 vial. DMF (3.00 mL) was added and the resulting mixture was stirred at 60 C. for 2.5 h. Control by LC/MS showed product.

[0406] The reaction mixture was diluted with water (40 mL) and extracted with MTBE (320 mL). The combined organic layers were washed with water (20 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0407] Yield: 43 mg (80%), white solid

Compound 33 (Comp-33)

N-(4-{2-[3-(4-hydroxyphenoxy)propyl]phenyl}butyl)-Nmethylethanediamide

[0408] Step 1:

[0409] N-{4-[2-(3-hydroxypropyl)phenyl]butyl}-N-methylethanediamide (BB-9, 400 mg) was suspended in CH2Cl2 (20 mL). PPh3 (610 mg) and CBr4 (771 mg) were added and the resulting mixture was stirred at r.t. for 30 min. Control by TLC (PE/EtOAc=1:1) and LC/MS showed complete conversion.

[0410] The reaction mixture was concentrated in vacuo and the residue was purified by preparative TLC (PE/EtOAc=1:1).

[0411] Step 2:

[0412] N-{4-[2-(3-bromopropyl)phenyl]butyl}-N-methylethanediamide (50 mg), Hydroquinone (77.5 mg) and K2CO3 (58.4 mg) were placed in a G16 vial. DMF (3.00 mL) was added and the resulting mixture was stirred at 60 C. for 1 h. Control by LC/MS showed product.

[0413] The reaction mixture was diluted with water (40 mL) and extracted with MTBE (320 mL). The combined organic layers were washed with water (20 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0414] Yield: 39 mg (72%), white solid

Compound 34 (Comp-34)

N-[4-(2-{3-[(4-hydroxyphenyl)methoxy]propyl}phenyl)butyl]-N-methylethanediamide

[0415] Step 1

[0416] 4-Hydroxybenzaldehyde (2.00 g) and imidazole (2.79 g) were dissolved in DMF (20.0 mL). TIPSCl (5.96 mL) was added and the resulting mixture was stirred at 60 C. for 2d.

[0417] Control by TLC (PE/EtOAc=95:5) and LC/MS showed complete conversion.

[0418] The reaction mixture was diluted with water (100 mL) and extracted with MTBE (340 mL). The combined organic layers were washed with 1N NaOH (30 mL) and brine (20 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=95:5).

[0419] Yield: 3.94 g (86%) pale yellow oil

[0420] Step 2

[0421] 4-{[tris(propan-2-yl)silyl]oxy}benzaldehyde (3.94 g) was dissolved in EtOH (30.0 mL) and cooled to 0 C. NaBH 4 (535 mg) was added and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 18 h.

[0422] Control by TLC (PE/EtOAc=8:2) and LC/MS showed product.

[0423] The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (340 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (PE/EtOAc=8:2).

[0424] Step 3

[0425] (4-{[tris(propan-2-yl)silyl]oxy}phenyl)methanol (300 mg) was dissolved in dry THF (5.00 mL). NaH (60% in mineral oil, 64.2 mg) was added and the mixture stirred at r.t. for 15 min. Allylbromide (231 L) was then added and the resulting mixture was stirred at r.t. for 2.5 h. Control by LC/MS showed complete conversion.

[0426] The reaction mixture was poured into water (30 mL) and extracted with EtOAc (310 mL). The combined organic layers were washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was used for further transformation as such.

[0427] Yield: 372 mg (crude) yellow oil

[0428] Step 4

[0429] {4-[(prop-2-en-1-yloxy)methyl]phenoxy}tris(propan-2-yl)silane (356 mg) was placed in a 10 mL flask and cooled to 0 C. under and Ar-atmosphere. 9-BBN (0.5 M in THF, 3.33 mL) was then added dropwise and the resulting mixture was stirred at 0 C. for 30 min and then at r.t. for 2 h.

[0430] A solution of Na2CO3 (147 mg) in water (3.00 mL) was then added and stirring was continued at r.t. for 30 min. Then N-[4-(2-iodophenyl)butyl]-N-methylethanediamide (BB-2, 100 mg) and PdCl2(PPh3)2 (9.7 mg) were added and the mixture heated to 50 C. for 2 h. Control by LC/MS showed product. The reaction mixture was cooled to r.t., diluted with water (20 mL) and the layers were separated. The aqueous layer was extracted with EtOAc (15 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was passed through a short column of SiO2 (PE/EtOAc 1:1). The still crude product was then used for further transformation as such.

[0431] Yield: 248 mg (crude) yellow oil.

[0432] Step 5

[0433] N-methyl-N-[4-(2-{3-[(4-{[tris(propan-2-yl)silyl]oxy}phenyl)methoxy]propyl}phenyl)butyl]ethanediamide (154 mg, crude IK-0357/20) was dissolved in THF (5.00 mL). TBAF*3H 2O (131 mg) was added and the resulting mixture was stirred at r.t. for 30 min. Control by LC/MS showed complete conversion.

[0434] The reaction mixture was poured into water (40 mL) and extracted with EtOAc (320 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0435] Yield: 76 mg (68%), white solid.

Compound 35 (Comp-35)

N[(5Z)-13-[(methanesulfonylcarbamoyl)methoxy]tridec-5-en-1-yl]-Nmethylethanediamide

[0436] 2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic acid (BB-4, 50.0 mg) was dissolved in THF (2.00 mL). 1,1-Carbonyldiimidazole (25.0 mg) was added and the resulting mixture was stirred at r.t. for 1.5 h. Then DBU (50.0 L) and methanesulphonamide (16.0 mg) were added and stirring was continued at r.t. for 18 h. Control by LC/MS showed product. The reaction mixture was diluted with water (30 mL) and extracted with CH 2Cl2 (320 mL). The combined organic layers were washed with 1N HCl (20 mL) and brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by preparative TLC (CH2Cl2/MeOH=95:5).

[0437] Yield: 27 mg (44%) white solid

Analytical Devices Used to Analyze Comp-14 to Comp-34

[0438] Analytical LC/ESI-MS: Waters 2700 Autosampler. Waters 1525 Multisolvent Delivery System. 5 L sample loop. Column, Phenomenex Onyx Monolythic C18 502 mm, with stainless steel 2 m prefilter. Eluent A, H.sub.2O+0.1% HCOOH; eluent B, MeCN. Gradient, 5% B to 100% B within 3.80 min, then isocratic for 0.20 min, then back to 5% B within 0.07 min, then isocratic for 0.23 min; flow, 0.6 ml/min and 1.2 ml/min.

[0439] Waters Micromass ZQ 4000 single quadrupol mass spectrometer with electrospray source. MS method, MS4_15 minPM-80-800-35V; positive/negative ion mode scanning, m/z 80-800 in 0.5 s; capillary voltage, 3.50 kV; cone voltage, 50 V; multiplier voltage, 650 V; source block and desolvation gas temperature, 120 C. and 300 C., respectively. Waters 2487 Dual A Absorbance Detector, set to 254 nm. Software, Waters Masslynx V 4.0.

[0440] Waters Micromass LCZ Platform 4000 single quadrupol mass spectrometer with electrospray source. MS method, MS4_15 minPM-80-800-35V; positive/negative ion mode scanning, m/z 80-800 in 1 s; capillary voltage, 4.0 kV; cone voltage, 30 V; multiplier voltage, 900 V; source block and desolvation gas temperature, 120 C. and 300 C., respectively. Waters 996 Photodiode Array Detector, set 200 to 400 nm. Software, Waters Masslynx V4.0.

[0441] Values for [M+H]+ given in the examples are those found within the corresponding LC/MS chromatogram for the respective compound. These values were all found within tolerable margins of +/0.3 units compared to calculated exact mass upon protonation of the compound.

[0442] Preparative thinlayer chromatography (preparative TLC): Merck PLC plates, silica gel 60 F254, 0.5 mm, 1.0 mm or 2.0 mm.

[0443] Column chromatography: Acros silica gel 60A, 0.035-0.070 mm.

[0444] Preparative HPLC-MS: Waters 2767 Autosampler, Waters 600 Multisolvent Delivery System with analytical pump heads (100 L); Waters 600 Controller; Waters 2525 Binary Gradient Modul with preparative pump heads (500 L). At-Column-Dilution: solvent1, MeCN:H2O 70:30 (v/v), solvent2, MeCN:MeOH:DMF 80:15:5 (v/v/v); flow rate, 5 mL/min. Autosampler 2767 with 10 mL syringe and 10 mL Sample loop. Column 6-position valve Flom 401 with Waters X-Terra RP18, 5 m, 19150 mm with X-Terra RP18 guard cartridge 5 m, 1910 mm, used at flow rate 20 mL/min; Waters SunFire Prep OBD 5 3050 mm with SunFire RP18 guard cartridge 5 m, 1910 mm, used at flow rate 25 mL/min; Waters Atlantis Prep T3 OBD 5 m, 3050 mm with Atlantis guard cartridge, used at flow rate 50 mL/min; Waters X-Bridge Prep OBD 5 m, 19150 mm with X-Bridge RP18 guard cartridge 5 m, 1910 mm used at flow rate 20 mL/min; Waters Atlantis Prep T3 OBD 5 m, 1950 mm with Atlantis guard cartridge, used at flow rate 25 mL/min and YMC-Actus Hydrosphere C18 5 m, 2050 mm with Actus guard cartridge, used at flow rate 20 mL/min. Eluent A, H.sub.2O containing 0.1% (v/v) HCOOH or H.sub.2O containing 0.1% (v/v) NEt3; eluent B, MeCN. Different linear gradients, individually adapted to sample. Injection volume, 9 mL, depending on sample. Make-up solvent, MeOH-MeCNH2OHCOOH 80:15:4.95:0.05 (v/v/v/v). Make-up pump, Waters Reagent Manager, flow rate 0.5 mL/min. Waters ZQ single quadrupole mass spectrometer with electrospray source. Positive or negative ion mode scanning m/z 105-950 in 1 s; capillary, 3.6 kV; cone voltage, 45 V; multiplier voltage, 700 V; probe and desolvation gas temperature, 120 C. and 250 C., respectively. Waters Fraction Collector 2767 with mass or UV-triggered fraction collection. Waters 2487 Dual A Absorbance Detector, set to 254 nm. Software, Waters Masslynx V 4.0 SP4.

[0445] 1H NMR spectra were recorded at room temperature on a Bruker Supraleitendes Fourier NMR Spektrometer, Avance 300 MHz. Chemical shifts 6 are reported in ppm. Multiplicity of a certain signal (singlet, doublet, triplet, quartet, multiplet) is indicated by the respective abbreviation (s, d, t, q, m respectively). br s indicates a broad singlet, mC a centered multiplet. The solvent residual signals were used as internal standards: (CDCl3)=7.26, (d6-DMSO)=2.50, (CD3OD)=3.31, (d6-acetone)=2.05.

TABLE-US-00019 TABLE xxx Calculated exact mass of Comp-14 to Comp-34 Calculated Compound IUPAC name Comp-00 exact mass M + 1 N-methyl-N-[(5Z)-13-[(1H-1,2,3,4-tetrazol-5-yl)methoxy]tridec-5-en-1- Comp-14 380.25359 381.260869 yl]ethanediamide N-(4-{2-[3-(Carbamoylmethoxy)propyl]phenyl}butyl)-N-methylethanediamide Comp-15 349.20016 350.207437 N-Methyl-N-[(5Z)-13-[(5-oxo-4,5-dihydro-1H-1,2,4-triazol-3- Comp-16 395.25326 396.260535 yl)methoxy]tridec-5-en-1-yl]ethanediamide N-methyl-N-[(5Z)-13-[(phenylcarbamoyl)methoxy]tridec-5-en-1- Comp-17 431.27841 432.285687 yl]ethanediamide N-methyl-N-[(5Z)-13-{[(oxan-4-yl)carbamoyl]methoxy}tridec-5-en-1- Comp-18 439.30462 440.311902 yl]ethanediamide N-methyl-N-[(5Z)-13-{[(1,3-oxazol-2-yl)carbamoyl]methoxy}tridec-5-en-1- Comp-19 422.25292 423.260201 yl]ethanediamide N-[(5Z)-13-{[(4-methoxyphenyl)carbamoyl]methoxy}tridec-5-en-1-yl]-N- Comp-20 461.28897 462.296252 methylethanediamide N-Methyl-N-[4-(2-{3- Comp-21 425.23146 426.238737 [(phenylcarbamoyl)methoxy]propyl}phenyl)butyl]ethanediamide N-Methyl-N-[4-(2-{3-[2-oxo-2-(pyrrolidin-1- Comp-22 403.24711 404.254387 yl)ethoxy]propyl}phenyl)butyl]ethanediamide N-Methyl-N-[4-(2-{3-[2-(morpholin-4-yl)-2- Comp-23 419.24202 420.249302 oxoethoxy]propyl}phenyl)butyl]ethanediamide N-[(5Z)-13-{[(benzenesulfonyl)carbamoyl]methoxy}tridec-5-en-1-yl]-N- Comp-24 495.24031 496.247589 methylethanediamide 4-{2-[3-(2-{4- Comp-25 469.22129 470.228567 [(methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetamido}benzoic acid N-[(5Z)-13-[2-(4-hydroxy-2-oxo-2,5-dihydrofuran-3-yl)-2-oxoethoxy]tridec-5- Comp-26 438.2366 439.243883 en-1-yl]-N-methylethanediamide N-[4-(2-{3-[2-(hydroxymethyl)phenoxy]propyl}phenyl)butyl]-N- Comp-27 398.22056 399.227838 methylethanediamide N-[(5Z)-13-(2-benzenesulfonamidoethoxy)tridec-5-en-1-yl]-N- Comp-28 481.26104 482.268324 methylethanediamide N-[(5Z)-13-(2-hydroxyphenoxy)tridec-5-en-1-yl]-N-methylethanediamide Comp-29 390.25186 391.259138 N-[(5Z)-13-(3-hydroxyphenoxy)tridec-5-en-1-yl]-N-methylethanediamide Comp-30 390.25186 391.259138 N-(4-{2-[3-(3-hydroxyphenoxy)propyl]phenyl}butyl)-N-methylethanediamide Comp-31 384.20491 385.212188 N-[(5Z)-13-(4-hydroxyphenoxy)tridec-5-en-1-yl]-N-methylethanediamide Comp-32 390.25186 391.259138 N-(4-{2-[3-(4-hydroxyphenoxy)propyl]phenyl}butyl)-N-methylethanediamide Comp-33 384.20491 385.212188 N-[4-(2-{3-[(4-hydroxyphenyl)methoxy]propyl}phenyl)butyl]-N- Comp-34 398.22056 399.227838 methylethanediamide N-[(5Z)-13-[(methanesulfonylcarbamoyl)methoxy]tridec-5-en-1-yl]-N- Comp-35 433.22466 434.231939 methylethanediamide

EXAMPLE 2: ANTI-ARRHYTHMIC EFFECT OF METABOLICALLY ROBUST ANALOGS OF 17,18-EEQ ON NRCMS

Materials and Methods

[0446] The structures of all compounds tested are given in FIG. 1. The compounds include analogues being part of the invention (Comp-01 to Comp-05), which were synthesized as described in example 1, and additional related compounds (Comp-06 to Comp-13). A 1000-fold stock solutions in ethanol of the test compounds was prepared before use.

[0447] In order to measure the biological activities of metabolically robust analogs of CYP-eicosanoids an established cell model was used (Kang, J. X. et al., 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. Indicating their anti-arrhythmic potential, test compounds reduce the spontaneous beating rate of these cells and prevent their irregular and asynchronous contraction in response to arrhythmic substances.

[0448] Isolation and cultivation of NRCMs were performed as described previously (Wallukat, G et al., J Clin Invest. 1999; 103: 945-952; Falck, J R et al., J Med Chem. 2011 Jun. 23; 54(12):4109-18). The isolated cells were cultured as monolayers on the bottom (12.5 cm2) of Falcon flasks in 2.5 ml of Halle SM 20-l medium equilibrated with humidified air. The medium contained 10% heat-inactivated FCS and 2 mol/l fluoro-deoxyuridine (Serva, Heidelberg, Germany), the latter to prevent proliferation of non-muscle cells. The NRCMs (2.4106 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. After four hours of incubation cells were adopted for 10 min to 31 C. and beating was recorded using an inverted microscope (Leica DM IRB) equipped with a CCD camera and coupled to an lonOptix (software: lonWizard6, lonOptix). 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 ( 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. Stock solutions of the compounds were prepared in ethanol and applied to give a final concentration of either 20 nM or 30 nM on NRCMs (n=6 per compound). The vehicle control (0.1%) showed no effect on the basal beating rate.

Results

[0449] The results are presented in FIG. 1. The potential of the 17,18-EEQ analogs to reduce the spontaneous beating rate ranged between 1.3+/1.0 delta bpm up to 38.0+/3.3 delta bpm according to distinct structural features. Free carboxylic acid derivatives showed a greater reduction than analogs where the carboxy group was esterified to a polyalkoxyalkyl or an amino acid (Comp-10 vs. Comp-07 and Comp-08 and Comp-06 vs. Comp-09). The results also showed, that it is possible to replace the double bond in 11,12-position with a phenyl ring without major loss of activity (Comp-06 and Comp-05). However, shifting the 11,12-double bond to the 14,15-position strongly reduced the negative chronotropic effect on NRCMs (Comp-06 vs. Comp-11). Moreover, analogs containing an 3-oxa group showed an equal potency to reduce the beating rate (Comp-06 vs. Comp-02 and Comp-03 or Comp-10 vs. Comp-01) or increased the negative chronotropic effect (Comp-10 vs. Comp-04). For the oxamide group it was shown that it was essential for the in vitro efficacy, since two degradation products of the oxamide group were inactive (Comp-06 vs. Comp-12 and Comp-13). As shown in FIG. 1 continued further compounds bearing both an 3-oxa and an oxamide group showed good activity (Comp-14 to Comp-35).

EXAMPLE 3: ANTI-ARRHYTHMIC EFFECT OF 17,18-EEQ AGONISTS COMP-02 ON ATRIAL FIBRILLATION

[0450] This example shows that the agonistic analog Comp-02 ameliorates atrial fibrillation

Materials and Methods

[0451] Study design: To gain insight into in-vivo effect of synthetic 17,18-EEQ-agonists, atrial fibrillation studies were performed in male B16 mice as described in Westphal, C. et al., PLoS ONE. 2013, 8(8): e73490. Briefly, moderate cardiac hypertrophy was induced by continuous infusion of isoproterenol via subcutaneously implanted osmotic minipumps (Alzet) at a rate of 40 mg/kg/d for two weeks. After two weeks of treatment, ECG and electrophysiological data were recorded. Programmed electrical stimulation (PES) was performed in the right atrium or right ventricle using a digital electrophysiology lab (EP Tracer; CardioTek) to determine refractory periods and arrhythmia inducibility. Atrial arrhythmias were defined as fast (>800 bpm) electrical activity in the right atrial electrograms, with ECG P waves different to normal sinus rhythm and subsequent fast, but physiological activation of the ventricles (ECG R wave and right ventricular electrograms similar to normal sinus rhythm). Atrial fibrillation was defined as fast, irregular activity in the right atrial electrograms with irregular conduction to the ventricles (high variability of RR intervals). Ventricular arrhythmias were defined by fast (>800 bpm) activity originating from the ventricular myocardium (change in morphology of ECG R waves and local right ventricular electrograms compared to normal sinus rhythm). During inhalation anesthesia with isoflurane (2% with 360 ml/min air flow; Univentor 400 anesthesia unit), the animals' body temperature was kept constant at 37 C. using a homeothermic blanket control unit (Hugo Sachs Elektronik, Harvard Apparatus) with rectal temperature control. After preparation of the right jugular vein, a 2 French octapolar electrophysiology catheter (CIBER mouse cath; NuMed) was placed in the right heart, including atrium and ventricle. PES was performed using a standardized protocol that included trains of 10 basal stimuli (S1) followed by up to 3 extra stimuli (S2-S4), delivered with a coupling interval decreasing in steps of 5 ms until ventricular or atrial refractoriness was reached. The stimulation procedures were repeated at three different basal cycle lengths (100 ms, 90 ms, 80 ms) with each animal. Occurrence and duration of inducible arrhythmias were documented. Only stimulation protocols with reproducible arrhythmias longer than five consecutive beats in ventricle and episodes longer than 350 ms in the atria were considered positive. Arrhythmia inducibility was calculated as the percentage of effective (positive) out of total protocols applied. Accordingly, the arrhythmia inducibility of individual animals could take a value of 0, 33, 66 or 100%. For statistical evaluation, the data obtained for the individual animals in a given group were averaged and are given as meanSEM. For scoring the severity of induced arrhythmias, three response categories were defined: sustained WO consecutive ventricular extrasystoles, VES or atrial fibrillation episodes 30 sec in at least one protocol), non-sustained (<10 VES or atrial fibrillation episodes <30 sec in at least one protocol) and no arrhythmias in all three protocols. The data are given as percentage of animals in a given group assigned to these categories.

Results

[0452] The results are presented in FIG. 2. Bolus injection of the synthetic 17,18-EEQ agonist (Comp. 02) did not induce any obvious negative side effects. Programmed electrical stimulation induced atrial fibrillation in a majority of vehicle-treated mice (n=12). A single i.v. injection of the synthetic 17,18-EEQ-agonist Comp-02 (2 mg/kg body weight) reduced significantly the sum of totally induced atrial fibrillation episodes (atrial fibrillation burden) (n=14); FIG. 2 A. Moreover, the severity of induced atrial fibrillation episodes was significantly reduced. In particular, the inducibility of sustained arrhythmia episodes was significantly reduced by 62%; FIG. 2 B.

EXAMPLE 4: ANTI-ARRHYTHMIC EFFECT OF 17,18-EEQ AGONISTS ON SPONTANEOUSLY VENTRICULAR TACHYCARDIAS IN ACUTE MYOCARDIAL INFARCTION

[0453] This example shows that the agonistic analog Comp-03 ameliorates arrhythmias as induced by myocardial infarction.

Materials and Methods

[0454] Study design: To gain insight into the in-vivo effects of synthetic 17,18-EEQ-agonists, myocardial infarction studies were performed in male Wistar rats. Briefly, rats weighing 220-250 g were randomized to receive an i.v. bolus of Comp-03 (100 g in 300 l 0.9% NaCl) or only 300 l 0.9% NaCl as vehicle control ten minutes before induction of myocardial infarction. For safe bolus application, animals were mildly anesthetized using isoflorane. Continuous monitoring of the surface-ECG was started (EPTracer, Netherlands) and maintained until the end of the study (45 minutes after induction of myocardial infarction). After recording of the basal ECG, myocardial infarction was induced by ligation of the left anterior descending artery (LAD). 45 minutes after myocardial infarction animals were sacrificed and organ harvested. Samples from urine, blood, liver, kidney and heart were stored for further analysis.

[0455] Method of arrhythmia analysis: Ventricular tachycardia burden was calculated as the sum of all arrhythmic events originating from the ventricular myocardium, which were observed within the first hour after induction of myocardial infarction. In order to quantify not only the frequency but also the severity of the ventricular arrhythmias, an arrhythmia-severity-score was calculated. This score was calculated as the sum of the number of different arrhythmia events (PVC, couplet, triplet, VT<1.5 sec, VT >=1.5 sec), each class factorized by an increasing severity index of 1-5 (e.g. PVC1, couplets2, triplets 1.5 sec3, VT>=1.5 sec5).

Results

[0456] The results are presented in FIG. 3. Bolus injection of the synthetic 17,18-EEQ agonist (Comp-03) did not induce any obvious negative side effects. Ventricular arrhythmias occurred after coronary artery ligation and were observed as single premature ventricular contractions (PVC), short runs of non-sustained ventricular tachycardia (VT) and ventricular tachycardia/fibrillation. Rats treated with the synthetic 17,18-EEQ-agonist (n=10) showed a significantly reduced ventricular tachycardia duration compared to controls (n=9); FIG. 3 A. Moreover, the arrhythmia severity score was significantly reduced. FIG. 3 B.

EXAMPLE 5: CARDIOPROTECTIVE EFFECT OF 17,18-EEQ IN ISCHEMIA/REPERFUSION DAMAGE OF THE HEART

[0457] This example shows that the agonistic analog Comp-03 ameliorates post-ischemic recovery as induced by a defined period of ischemia.

Materials and Methods:

[0458] Study design: To gain insight into the cardioprotective effects of synthetic 17,18-EEQ-agonists, ischemia-reperfusion studies were performed in isolated mouse hearts. Briefly, hearts were perfused in the Langendorff mode as described in Seubert et al., Circ Res. 2004; 95:506-514. Hearts were perfused with buffer for a 10-minutes stabilization period, then perfused with either the agonistic analog Comp-03 (1 M) or vehicle for 10 minutes, then subjected to 35 minutes global no-flow ischemia, followed by 40 minutes reperfusion. During ischemia and reperfusion the infusion with either the agonistic analog Comp-03 or vehicle was kept constant. Recovery of contractile function was taken as left ventricular developed pressure (LVDP) at the end of reperfusion expressed as a percentage of pre-ischemic LVDP.

Results

[0459] The results are presented in FIG. 4. Continuous infusion of 1 M of the synthetic 17,18-EEQ agonist (Comp-03) did not induce any obvious negative side effects. Immediately after the 35 minutes episode of global ischemia contractility of the control hearts (n=14) as expressed by the left ventricular developed pressure (LVdP) were strongly reduced and returned then gradually in the reperfusion phase to about 50% of the pre-ischemic values. Hearts treated with the synthetic 17,18-EEQ-agonist Comp-03 (n=14) showed significantly improved recovery of contractility.

EXAMPLE 6: IN VITRO INHIBITION OF RECOMBINANT HUMAN SOLUBLE EPOXIDE HYDROLASE

Materials and Methods

[0460] A selection of compounds was tested for the ability to inhibit human soluble expoxide hydrolase (sEH). The metabolically robust 17,18-EEQ analogs themselves are not prone to metabolization by the sEH but might act as inhibitors of this enzyme. Briefly, the assay was performed at 37 C. for 20 min in a final volume of 100 L potassium phosphate buffer (0.1 M, pH 7.2) containing 50 M 14,15-EET as substrate. The reactions were started by adding enzyme (107.5 ng per reaction, 12.06 U/ml activity, Cayman Chemicals) and terminated with 300 l ethyl acetate. The remaining substrate and its product (14,15-DHET) were extracted and analyzed by reversed-phase high performance liquid chromatography (RP-HPLC) (Muller, D. N. et al., Biochem J 2007. 403(1): p. 109-118). Metabolically robust analogs shown in FIG. 5 were tested at final concentration of 10 M for potency of sEH-inhibition compared to vehicle control (1% DMSO), n=2-4.

Results

[0461] FIG. 5 shows that some of the tested 17,18-EEQ analogs inhibited human sEH up to 76.6%. These compounds share one structural feature, containing an urea group (Comp-01, -07, -08 and Comp-10). In contrast, both compounds tested containing an oxamide group did not show sEH-inhibition (Comp-02 and Comp-03).

EXAMPLE 7: PERMEABILITY POTENTIAL OF METABOLICALLY ROBUST ANALOGS OF CYP-EICOSANOIDS TESTED IN CACO-2 CELLS

Materials and Methods

[0462] To predict human intestinal permeability and to investigate drug efflux, parameters affecting the bioavailability of a compound (van Breemen R B & Li Y., Expert Opin Drug Metab Toxicol. 2005 August; 1(2):175-85.), a selection of metabolically robust 17,18-EEQ analogs were tested for their potential to permeate through a confluent monolayer of Caco-2 cells (a human colon adeno-carcinoma cell line). Stock solutions of compounds were generated in DMSO (1 mM) and compounds were tested at a final concentration of 1 M. A solution of a test compound placed on the apical side of a Caco-2 cell monolayer (2 h incubation time), and the rates of appearance of the test compounds on the basolateral side of the cells are measured to assess the permeability of the monolayer and are given as permeability from A to B in cm/s for the tested compounds (FIG. 6).

Results

[0463] The results presented in FIG. 6 show that compounds with an urea group showed less permeability (Comp-01 and Comp-04) than compounds containing an oxamide group (Comp-02 and Comp-03). An additional polyalkoxyalkyl esterified to the carboxylic acid group further improved permeability (Comp-02 vs. Comp-03).

EXAMPLE 8: ABILITY OF METABOLICALLY ROBUST ANALOGS OF CYP-EICOSANOIDS TO INCORPORATE INTO PHOSPHOLIPID MEMBRANES

Material and Methods

[0464] To see if modified structures (selected metabolically robust analogs of CYP-eicosanoids, Table 1) are able to incorporate into phospholipid membranes, a cardiomyocyte cell line (H9c2, rat) was incubated with test compounds for four hours at a final concentration of 1 M (n=2). After incubation cellular lipids were extracted with a chloroform/methanol 1:2 mixture. To differentiate between free and incorporated compounds an aliquot of the samples was subjected to alkaline hydrolysis. Both extracts were analyzed for the test compounds using LC-MS/MS. 17,18-EEQ and 20-HETE (Kaduce T L et al., J Biol Chem. 2004 Jan. 23; 279(4):2648-56.) served as positive control for membrane incorporation.

Results

[0465] FIG. 7 shows that in contrast to the positive controls (17,18-EEQ and 20-HETE) the compounds of invention (Comp-02 and Comp-04) as well as the oxamide containing compound (Comp-06) showed no incorporation into phospholipid membranes. The urea Comp-10 lacking the 3-oxa group showed weak incorporation.

EXAMPLE 9: SOLUBILITY ASSESSMENT OF METABOLICALLY ROBUST ANALOGS OF 17,18-EEQ

Materials and Methods

[0466] Aqueous solubility of Comp-06 and Comp-02 was determined in deionised water at 37 C. Known amounts of the compounds were suspended by agitation for 24 hours. The filtrates were isolated and content of the appropriate compounds was determined by HPLC analysis.

Results

[0467] In Table 1 it is shown, that the introduction of an 3-oxa group into the structure improved solubility. Comparing the solubility in deionised water, solubility of Comp-02 was approximately 10 greater than that of Comp-06.

TABLE-US-00020 TABLE 1 OMT-02 OMT-06 Highest and Average OMT-02 Highest and lowest Average OMT-06 Aqueous lowest OMT-02 solubility at OMT-06 solubility solubility at media, solubility values 37 C., values at 37 C., 37 C., 0.1M at 37 C., mg/ml mg/ml mg/ml mg/ml Deionised 0.075 0.068 0.0070 0.0068 water, pH 0.059 0.0066 5.90

EXAMPLE 10: PHARMACOKINETIC PROPERTIES OF METABOLICALLY ROBUST ANALOGS OF 17,18-EEQ

Material and Methods

[0468] Study Design:

[0469] To gain insight into pharmacological properties of synthetic 17,18-EEQ agonists pharmacokinetic studies were performed with two selected compounds (Comp-02 and Comp-06). These compounds were pharmacologically evaluated in plasma after single intravenous and oral administration. Therefore, male C57BL/6 mice (Janvier Labs (France), n=12 per group) were administered to the test item at a dose of 2 mg/kg (i.v.) and 8 mg/kg (p.o.), respectively. Intravenous application of Comp-02 (sodium salt in isotonic salt solution) and Comp-06 (free acid in DMSO/PEG400 20:80) was performed via the tail vein, and for oral administration via gavage. To assure equal solubility Comp-06 was formulated in DMSO/PEG400 20:80, since Comp-06 showed impaired solubility as sodium salt. Blood sampling was of 100 l were obtained at two different time point for each mouse (0.083/2 h, 0.25/4 h, 0.5/8 h and 1/24 h). Biological samples were subjected to an extraction procedure (40 l Acetonitril+22.4 l sample, shaking, 10 min centrifugation at 6000g at room temperature, dilution of 50 l supernatant 1:1 with water). For analysis 20 l of sample were injected into the LC-MS, Accela 1250 UHPLC system, Accela Open Autosampler, Q-Exactive mass spectrometer (Thermo Fisher Scientific). Appropriate samples were used as controls: zero sample, calibration standard and QC sample. The pharmacokinetic analysis were performed applying a non-compartment model using the Kinetica 5.0 software (Thermo Scientific, Waltham, USA). All given parameters were obtained by trapezoid area calculation.

TABLE-US-00021 Cmax (ng/ml) maximal observed plasma concentration tmax (h) time of maximal observed plasma concentration AUC0-(ng*h/ml) area under the concentration-time curve extrapolated to infinity F (%) oral bioavailability expressed as percentage

Results

[0470] The results presented in Table 2 show improved properties in pharmacokinetic parameters of Comp-02 compared to Comp-06. The compound of invention (Comp-02) differs only in an additional 3-oxa group, which is missing in Comp-06. This structural feature greatly improved the oral bioavailability (87% vs 5.3%). Moreover, the maximal observed concentration was much higher and reached faster with Comp-02 (903.3 ng/ml and 0.3 h) than Comp-06 (24.4 ng/ml and 0.5 h). Further, the AUC0-, as measure for total compound exposure over time, was higher with Comp-02 (1818 ng*h/ml) compared to Comp-06 (31 ng*h/ml).

TABLE-US-00022 TABLE 2 Comp-02 Comp-06 Iv po iv po 2 mg/kg 8 mg/kg 2 mg/kg 8 mg/kg C max (ng/ml) 903.8 24.4 t max (h) 0.3 0.5 AUC0-(ng*h/ml) 524 1818 146 31 F (%) 100 87 100 5.3

EXAMPLE 11: CARDIOPROTECTIVE EFFECT OF SYNTHETIC 17,18-EEQ-AGONIST IN ISCHEMIA/REPERFUSION OF THE HEART

[0471] Further to the example 5, this example shows that another agonistic analog of the invention, namely Comp-02 ameliorates post-ischemic recovery as induced by a defined period of ischemia.

Materials and Methods

[0472] Study design: To gain insight into the cardioprotective effects of synthetic 17,18-EEQ-agonists, ischemia-reperfusion studies were performed in isolated mouse hearts of 12-14 week old male C57BL6/n animals. Briefly, mice were anesthetized with i.p. injected pentobarbital/heparin, hearts were rapidly removed and perfused in a Langendorff apparatus. Excised hearts were put in ice-cold Krebs-Henseleit buffer and aortas were cannulated. Modified Krebs-Henseleit buffer for retrograde perfusion contained 118 mM NaCl, 3.5 mM KCl, 1.3 mM MgSO.sub.4, 2.5 mM CaCl.sub.2, 24.7 mM NaHCO.sub.3, 1.4 mM KH.sub.2PO.sub.4 and 11 mM glucose, was aerated with 95% O.sub.2-5% CO.sub.2 and kept at constant temperature of 37 C. and pH 7.3. Perfusion pressure was adjusted to 80 mmHg and a latex balloon connected to a pressure transducer was inserted in the left ventricle through the left atrium. Hearts were perfused with buffer for a 15 minutes stabilisation period, followed by 10 minutes perfusion with either the agonistic analog Comp-02 (100 nM) or vehicle (0.9% NaCl in water). Afterwards hearts were subjected to 35 minutes global no-flow ischemia followed by 40 minutes of reperfusion. Compound or vehicle perfusion was kept constant during the reperfusion time. To analyse recovery of contractile function the left ventricular developed pressure (LVdP) at different time points during reperfusion was measured and expressed as percentage of pre-ischemic LVdP. Exclusion criteria for the hearts were: (i) a LVdP below 60 mmHg at the end of the stabilisation phase (baseline measurement), (ii) a heart rate below 260 bpm, (iii) coronary flow below 1.5 or over 4 ml/min and finally (iv) severe sustained arrhythmias during the stabilisation phase. The results are presented in FIG. 1 and expressed as meanSEM percentage of pre-ischemic LVdP. Data were analysed by two-tailed unpaired Student's t test and considered significant if p<0.05*.

Results

[0473] FIG. 8 shows that continuous infusion of 100 nM of Comp-02 did not induce any obvious negative side effects. After global ischemia, contractility of the control hearts (n=5) was strongly reduced. After 10 min of reperfusion time, pre-ischemic LVdP values ware at 1% but returned gradually to about 18% after 40 minutes reperfusion. In contrast, hearts treated with Comp-02 (n=5) showed significantly improved post-ischemic recovery of contractility function compared to the control group. Early (10 minutes) as well as late (40 minutes) time points during reperfusion phase showed better functional recovery with 18% and 59% of the pre-ischemic LVdP values, respectively.

EXAMPLE 12: CARDIOPROTECTIVE EFFECT OF ROBUST ANALOGS OF 17,18-EEQ ON ISOLATED PRIMARY CARDIOMYOCYTES

Materials and Methods

[0474] In vitro ischemic injury was induced in mouse neonatal primary cardiomyocyte culture by oxygen-glucose deprivation for 18 hours and a 24 hours reoxygenation period in a humidified incubator with 5% CO.sub.2/95% air. Cardiomyocytes were incubated with the test compounds during 4 hours prior to the oxygen glucose depriviation (OGD) induction. The test compounds were also present during the OGD insult and during the 24 h-reoxygenation period. After the reoxygenation period, cell number was assessed by Hoechst 33342 staining, apoptosis was measured by determining caspase 3/7 activation, and cell membrane integrity or necrosis was measured as LDH release.

Data were expressed as meanSD of three separate wells and individual comparisons were made with Student's t-test (SigmaPlot 9.0 program). The results were normalized to the normoxia-treated cells. Statistical significance (*) p<0.05 as compared to normoxia/DMSO-treated cells and () hypoxia/DMSO treated cells.

Results

[0475] FIG. 9A to 9B show, that Comp-02 partially protected against OGD-induced damage in primary cardiomyocytes. It ameliorates loss of cell number (by 35% with 30 nM or by 44% with 10 M) and reduced OGD-induced apoptosis and necrosis (by 43% and 29% respectively). The natural precursor of the compound of invention 17,18-EEQ appeared less efficacious, as it did not significantly affect cell number (at 30 nM and 10 M), and reduced apoptosis only at high concentration (10 M, 43%) but not at the low concentration tested (30 nM). However, to prevent OGD-reoxygenation induced cellular necrosis, as tested by LDH-release, Comp-02 and 17,18-EEQ seemed similarly effective. Comp-02 reduced cellular necrosis by 43% (30 nM) and by 45% (10 M) respectively. Comparably, 17,18-EEQ reduced LDH-release by 41% (30 nM) and by 37% (10 M) respectively, see FIG. 9C