U.S. patent application number 16/089461 was filed with the patent office on 2019-04-25 for analogs of cyp-eicosanoids for use in treating or preventing a disorder associated with neovascularization and/or inflammation.
The applicant listed for this patent is MAX DELBRUCK-CENTRUM FUR MOLEKULARE MEDIZIN, OMEICOS THERAPEUTICS GMBH. Invention is credited to Robert FISCHER, Anne KONKEL, Janine LOSSIE, Dominik MULLER, Wolf-Hagen SCHUNCK, Tim WESSER.
Application Number | 20190117597 16/089461 |
Document ID | / |
Family ID | 65863570 |
Filed Date | 2019-04-25 |
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United States Patent
Application |
20190117597 |
Kind Code |
A1 |
FISCHER; Robert ; et
al. |
April 25, 2019 |
ANALOGS OF CYP-EICOSANOIDS FOR USE IN TREATING OR PREVENTING A
DISORDER ASSOCIATED WITH NEOVASCULARIZATION AND/OR INFLAMMATION
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) for use in treating or reducing the risk of developing
or preventing: (i) neovascularization and/or (ii) inflammatory
disorder, in particular, ophthalmic disorders associated with
neovascularization and/or inflammation.
Inventors: |
FISCHER; Robert; (Berlin,
DE) ; SCHUNCK; Wolf-Hagen; (Berlin, DE) ;
MULLER; Dominik; (Berlin, DE) ; WESSER; Tim;
(Berlin, DE) ; KONKEL; Anne; (Berlin, DE) ;
LOSSIE; Janine; (Berlin, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
OMEICOS THERAPEUTICS GMBH
MAX DELBRUCK-CENTRUM FUR MOLEKULARE MEDIZIN |
Berlin
Berlin |
|
DE
DE |
|
|
Family ID: |
65863570 |
Appl. No.: |
16/089461 |
Filed: |
April 3, 2017 |
PCT Filed: |
April 3, 2017 |
PCT NO: |
PCT/EP2017/057830 |
371 Date: |
September 28, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62317253 |
Apr 1, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/4196 20130101;
A61K 31/40 20130101; A61P 29/00 20180101; A61K 31/192 20130101;
A61P 27/02 20180101; A61K 31/421 20130101; A61K 31/351 20130101;
A61K 31/5375 20130101; A61K 31/22 20130101; A61K 31/165 20130101;
A61K 31/341 20130101 |
International
Class: |
A61K 31/165 20060101
A61K031/165; A61K 31/192 20060101 A61K031/192; A61K 31/22 20060101
A61K031/22; A61K 31/341 20060101 A61K031/341; A61K 31/351 20060101
A61K031/351; A61K 31/40 20060101 A61K031/40; A61K 31/4196 20060101
A61K031/4196; A61K 31/421 20060101 A61K031/421; A61K 31/5375
20060101 A61K031/5375; A61P 27/02 20060101 A61P027/02; A61P 29/00
20060101 A61P029/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 26, 2017 |
EP |
17153412.6 |
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.n--O--(CH.sub.2).sub.k--X (II) wherein n is 0 or
an integer of from 3 to 8; and k is 0, 1 or 2; preferably with the
proviso that when n is 0 k is 1, most preferably k is 1; X
represents CH.sub.2OH, CH.sub.2OAc, CH(0) or a group selected from
the group consisting of: ##STR00055## preferably X is ##STR00056##
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(C.dbd.O)R.sup.11;
R.sup.11 is a C.sub.1-C.sub.6alkyl group which is optionally
substituted with one or more fluorine or chlorine atom(s); or 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.2--CH.sub.2).sub.i--R.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: ##STR00057## 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(.dbd.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(.dbd.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(.dbd.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(.dbd.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(.dbd.O)C.sub.11-C.sub.21alkyl; or
--C(.dbd.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 ##STR00058## 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(.dbd.O) by an amide bond; o is an
integer of from 1 to 10; R.sup.4 is selected from the group
consisting of: ##STR00059## h is 0, 1, or 2; R.sup.5 represents a
hydrogen atom; a fluorine or chlorine atom; --CF.sub.3;
--C(.dbd.O)OR.sup.51; --NHC(.dbd.O)R.sup.52;
--C(.dbd.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(C.dbd.O)(C.sub.1-C.sub.6)alkyl
group, or a
--O(C.sub.1-C.sub.6)alkyldiylO(C.dbd.O)(C.sub.2-C.sub.6)alkenyl
group; wherein the C.sub.1-C.sub.6alkyl group and the
C.sub.2-C.sub.6alkenyl group 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:
##STR00060## 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(.dbd.O)-aryl, --C(.dbd.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 C--OH; 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): ##STR00061## 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(.dbd.O)-aryl,
--C(.dbd.O)C.sub.1-C.sub.6alkyl, or
--SO.sub.2(C.sub.1-C.sub.6alkyl); or --SO.sub.2aryl; wherein any of
the foregoing C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, or aryl
are optionally substituted with one, two or three substituents
independently selected from the group consisting of --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N(C.sub.1-C.sub.6)dialkyl,
C.sub.1-C.sub.6alkylcarbonyloxy-,
C.sub.1-C.sub.6alkoxycarbonyloxy-,
C.sub.1-C.sub.6alkylcarbonylthio-,
C.sub.1-C.sub.6alkylaminocarbonyl-,
di(C.sub.1-C.sub.6)alkylaminocarbonyl-, fluorine or chlorine atom,
and 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,
--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; l is --(CH.sub.2).sub.m--Y, 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
--U--V--W--(CH.sub.2).sub.p--(CH.sub.3).sub.q, wherein p is an
integer from 0 to 6; q is 0 or 1; U is absent or selected from the
group consisting of CH, CH.sub.2 and NR.sup.40, with the proviso
that U is only CH if it forms an epoxy group together with V and W;
V is selected from the group consisting of --C(O)--,
--C(O)--C(O)--, --O--, and --S--; W is selected from the group
consisting of CH, CH.sub.2 and NR.sup.40 with the proviso that W is
only CH if it forms an epoxy group together with U and V; or Y
represents a group selected from the group consisting of:
##STR00062## 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(.dbd.O)aryl, or
--C(.dbd.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)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; f is an integer of from 0 to 2; with the proviso that
when X does not comprise a --C(.dbd.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 for use in treating,
reducing the risk of developing or preventing a disorder associated
with neovascularization and/or inflammation.
2. The compound for use according to claim 1, with the proviso that
when X does not comprise a --C(.dbd.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.
3. The compound for use according to claim 1, with the 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.3--O--(CH.sub.2)--X.sup.81;
--(CH.sub.2).sub.5--O--(CH.sub.2)--X.sup.81; wherein X.sup.81 x
represents a group selected from the group consisting of:
##STR00063## R.sup.1' is defined as R.sup.1 above; R.sup.2'
represents --NHR.sup.3'; --OR.sup.22';
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.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:
##STR00064## 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.
4. The compound according to claim 1, wherein X is ##STR00065##
wherein R.sup.2 is --OR.sup.22;
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.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:
##STR00066## 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.
5. The compound according to claim 1, wherein X is --C(.dbd.O)OH or
a suitable salt of the carboxylic acid, preferably a free
carboxylic acid.
6. The compound according to claim 1, wherein Y is one of the
oxamides defined according to claim 1.
7. The compound according to claim 1, wherein X is ##STR00067##
wherein R.sup.2 is --OR.sup.22;
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.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:
##STR00068## 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.
8. The compound according to claim 1, wherein X is C(.dbd.O)OH,
preferably the free carboxylic acid, and Y is one of the oxamides
defined according to claim 1.
9. The compound according to claim 1, with the formula (V): formula
(V): ##STR00069## wherein R.sup.55 represents --OH; --OR.sup.22;
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.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: ##STR00070## 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 0, 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.
10. The compound according to claim 9, wherein R.sup.55 represents
--OH or --(OCH.sub.2--CH.sub.2).sub.i--R.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.
11. The compounds according to claim 1, wherein the compound is
selected from the group consisting of: ##STR00071## ##STR00072##
##STR00073## ##STR00074## or a pharmaceutically acceptable salt
thereof.
12. The compound according to claim 1, with the formula (VI)
##STR00075## or a pharmaceutically acceptable salt thereof.
13. The compound for use according to claim 1, wherein the disorder
is associated with inflammation, preferably the disorder is
selected from inflammatory disorders, inflammation caused by other
diseases whatever type, etiology or pathogenesis, inflammation
caused by inflammatory diseases and immunological disorders.
14. The compound for use according to claim 1, wherein the disorder
associated with neovascularization and/or inflammation is an
ophthalmological disorder, preferably associated with
neovascularization, preferably age-related macular degeneration,
more preferably neovascular (wet) AMD, or atrophic (dry) AMD, or
retinopathy, more preferably retinopathy of prematurity (ROP);
diabetic retinopathy; diabetic proliferative retinopathy; retinal
vein occlusion, e.g. branch retinal vein occlusion, central retinal
vein occlusion; sickle cell retinopathy; and radiation retinopathy;
Best's disease or Stargardt's disease.
15. The composition for use according to claim 13, wherein the
compound or composition is administered orally, topically,
subcutaneously, intramuscularly, intravenously, intranasally,
intraocular, preferably orally or intraperitoneally, more
preferably orally.
16. The compound or composition for use 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.
Description
[0001] 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) for use in treating or reducing the risk of
developing or preventing: (i) neovascularization and/or (ii)
inflammatory disorder, in particular, ophthalmic disorders
associated with neovascularization and/or inflammation.
BACKGROUND OF THE INVENTION
[0002] Omega-6 and omega-3 polyunsaturated fatty acids (n-6 and n-3
PUFAs) are essential components of the mammalian diet. Biologically
most important n-3 PUFAs are eicosapentaenoic acid (EPA, 20:5 n-3)
and docosahexaenoic acid (DHA, 22:6 n-3). Dietary n-3 PUFAs have
effects on diverse physiological processes impacting normal health
and chronic disease, such as the regulation of plasma lipid levels,
cardiovascular and immune function, inflammation, insulin action,
and neuronal development and visual function.
[0003] Ingestion of n-3 PUFA will lead to their distribution to
virtually every cell in the body with effects on membrane
composition and function, eicosanoid synthesis, and signaling as
well as the regulation of gene expression.
[0004] Epidemiological and experimental studies showed that n-3
PUFA consumption is associated with a reduced risk of macular
degeneration. A major common mechanism in protecting against
macular degeneration and cancer consists in the capacity of n-3
PUFAs to inhibit pathological angiogenesis. EPA and DHA inhibit
abnormal retinal neovascularization, vascular permeability, and
inflammation. Angiogenesis is an essential step in tumor growth and
metastasis that is promoted by n-6 PUFAs and n-6 PUFA-derived
metabolites but inhibited by n-3 PUFAs and n-3 PUFA-derived
metabolites.
[0005] Simopoulos and colleagues summarized animal experiments and
clinical intervention studies indicating that n-3-PUFAs have
anti-inflammatory properties and, therefore, might be useful in the
management of inflammatory and autoimmune diseases (Simopoulos AP.
Omega-3 fatty acids in inflammation and autoimmune diseases. J Am.
Coll. NutL 2L 495-505 (2002)). Among the n-3-PUFAs, EPA and DHA
play an important and potent role with regard to anti-inflammatory
effects. (Calder C. P., Marine omega-3 fatty acids and inflammatory
processes: Effects, mechanisms and clinical relevance, Biochimica
et Biophsica Acta--Molecular and Cell Biology of Lipids, Volume
1851 (4), April 2015, 469-484).
[0006] Koto and colleagues showed that EPA has anti-inflammatory
activity in a mouse model for choroidal neovascularization (CNV)
(Koto et al. Eicosapentaenoic Acid Is Anti-Inflammatory in
Preventing Choroidal Neovascularization in Mice. Invest Ophthalmol
Vis Sci. 2007; 48:4328-4334). They demonstrated that an EPA-rich
diet results in significant suppression of CNV-related inflammatory
molecules in vivo and in vitro such as ICAM-1 and MCP-1 in
endothelial cells and VEGF and IL-6 in macrophages. Yanai and
colleagues demonstrated that dietary enrichment with n-3 PUFAs
suppresses choroidal neovascularization in a mouse model of
age-related macular degeneration (AMD) (Yanai et al. Cytochrome
P450-generated metabolites derived from .omega.-3 fatty acids
attenuate neovascularization. Proc Natl Acad Sci USA. 2014 Jul. 1;
111(26):9603-8.). Furthermore, they have shown that n-3 PUFAs have
anti-inflammatory properties in this model. This has been shown by
significantly reduced systemic immune-cell recruitment and down
regulation of Icam-1 and E-selectin expression on endothelial cells
and the ICAM-1 ligand CD11b-CD18 on the surface of circulating
leukocytes. N-3 PUFAs result also in suppression of macrophage
invasion into CNV lesions. They have further shown that this effect
is mediated by CYP-generated bioactive lipid mediators derived from
.omega.-3 LCPUFAs, specifically by the major CYP epoxygenase
metabolites derived from EPA (17,18-EEQ) and DHA (19,20-EDP) (Yanai
et al. Cytochrome P450-generated metabolites derived from .omega.-3
fatty acids attenuate neovascularization Proc Natl Acad Sci USA.
2014 Jul. 1; 111(26):9603-8 and WO 2014/110261 A1).
[0007] Laser induced CNV model in mice is a widely accepted model
for testing potential drugs for their effectiveness in treating
ophthalmic disorders associated with neovascularization and/or
inflammation, in particular AMD. Furthermore, ocular neovascular
diseases such as AMD are also suspected to have a significant
inflammatory component (Lopez et al. Pathologic features of
surgically excised subretinal neovascular membranes in age-related
macular degeneration. Am J Ophthalmol. 1991; 112(6):647-656.;
Grossniklaus et al. Macrophage and retinal pigment epithelium
expression of angiogenic cytokines in choroidal neovascularization.
Mol Vis. 2002 Apr. 21; 8:119-26; Lopez et al. Transdifferentiated
retinal pigment epithelial cells are immunoreactive for vascular
endothelial growth factor in surgically excised age-related macular
degeneration-related choroidal neovascular membranes. Invest
Ophthalmol Vis Sci. 1996, 37(5):855-868; Tezel et al. Pathogenesis
of age-related macular degeneration. Trends Mol Med. 2004;
10(9):417-420.; Schlingemann RO--Role of growth factors and the
wound healing response in age-related macular degeneration. Graefes
Arch Clin Exp Ophthalmol. 2004; 242(1):91-101). Grossniklaus and
colleagues have shown in choroidal membrane specimen from AMD
patients that progression of CNV represents a dynamic process with
not only angiogenesis but also with a strong inflammatory component
in particular macrophages. Based on the work by Ambati and
colleagues (Ambati et al. An animal model of age-related macular
degeneration in senescent Ccl-2- or Ccr-2-deficient mice. Nat. Med.
2003, 9, 1390-1397) linking AMD pathogenesis to complement system
and macrophages the inflammatory component of AMD was further
deciphered and changed the understanding of AMD pathogenesis
dramatically. Afterwards many research groups further explored the
role of inflammatory processes in AMD and CNV pathogenesis such as
recent work analyzed the macrophage polarization in experimental
and clinical choroidal neovascularization (Yang et al. Macrophage
polarization in experimental and clinical choroidal
neovascularization. Sci Rep., 2016 Aug. 4, 6:30933). Recent reviews
by the group of David Hinton, a pioneer of CNV research, and Campa
et al. (Inflammatory mediators and angiogenic factors in choroidal
neovascularization: pathogenetic interactions and therapeutic
implications. Mediators of Inflammation, 2010) nicely summarizes
the impact of inflammation in AMD pathogenesis and resulting
fibrosis (Ishikawa et al. Molecular mechanisms of subretinal
fibrosis in age-related macular degeneration. Exp Eye Res. 2016
January, 142:19-252016) and concluding CNV causing a heterogeneous
disease affecting the posterior segment of the eye which is more
properly definable as an aberrant tissue invasion of endothelial
and inflammatory cells, in which both angiogenesis and inflammation
are involved.
[0008] It becomes obvious that the laser-induced CNV model which is
widely used to elucidate the pathobiology of choroidal angiogenesis
and to identify novel therapeutic applications (Grossniklaus et al.
2010) has a strong inflammatory stimulus due to the injury after
laser burn of Bruch's membrane. Therefore, beside anti-angiogenic
compounds, many anti-inflammatory compounds showed broad activity
and the model can be also considered as a model for ocular
inflammation and treatment response and a model of ocular
inflammation would be considered by the skilled person to be
evidence of the suitability of a compound in the treatment of
inflammation as such.
[0009] During inflammation, circulating monocytes increasingly
leave the circulation and migrate into tissues where, following
conditioning by local growth factors, pro-inflammatory cytokines
and microbial products, they differentiate into macrophage or
dendritic cell populations. In the rat model shown in Example 5,
this event is visible by staining the infiltrated ED1-positive
monocytes/macrophages in heart and kidney slices with a respective
antibody. In general, recruitment of monocytes is essential for
effective control and clearance of viral, bacterial, fungal and
protozoal infections, but recruited monocytes also contribute to
the pathogenesis of inflammatory and degenerative diseases (Shi C.,
et al., Monocyte recruitment during infection and inflammation. Nat
Rev Immunol. 2011 Oct. 10; 11 (11) 762-74). Beyond promoting
atheroclerosis, recruited monocytes/macrophages are known to
contribute to acute and chronic inflammatory diseases of the heart
and kidney (Ingersoll et al., Monocyte trafficking in acute and
chronic inflammation. Trends Immuno 2011 October. 32(10) 470-7;
Hansson G., Inflammation, Atherosclerosis, and Coronary Artery
Disease. New Engl Jour Med 2005, (352) 1685-95; Kinsey et al,
Imflammation in Acute Kidney Injury. Experim Nephro 2008, (109)
e102-e107; Bonventre, J. Cellular pathophysiology of ischemic acute
kidney injury. J Clinic Invest 2011 November, (121) 4210-4221;
Guiteras R., et al., Macrophage in chronic kidney disease. Cli Kid
j 2016, vol. 9, no 6, 765-771).
[0010] The production of TNF-alpha plays an important role in
chronic inflammatory conditions, intermediary metabolism and
cardiovascular risk (Popa C. et al., The role of TNF-alpha in
chronic inflammatory conditions, intermediary metabolism, and
cardiovascular risk. J Lipid Res 2007, (48) 752-761). Aberrant
TNF-alpha production and TNF receptor signaling have been
associated with the pathogenesis of several chronic inflammatory
diseases (Parameswaran N. et al., Tumor Necrosis Factor-.alpha.
Signaling in Macrophages. Crit Rev Eukaryot Gene Expr 2010 20(2)
87-103). TNF-alpha has both diverse and potentially conflicting
roles in cardiac function and pathology (Sack M., Tumor necrosis
factor-alpha in cardiovascular biology and the potential role for
anti-tumor necrosis factor-alpha therapy in heart disease.
Pharmacol Ther 2002 April-May, 94(1-2) 123-135). As also shown in
Example 4, TNF-alpha can be produced in response to
pro-inflammatory stimuli by cardiomyocytes themselves. After
release into the surrounding tissue, TNF-alpha together with a wide
range of further mediators triggers leucocyte activation and
recruitment (Ghigo A. et al., Myocyte signalling in leucocyte
recruitment to the heart. Cardicovasc Res 2014 May. 102(2)
270-280).
[0011] One of the PUFAs most important biological roles is to
supply precursors for the production of bioactive fatty acid
metabolites that can modulate many functions. For instance,
arachidonic acid (AA; 20:4, n-6) is metabolized by Cytochrome P450
(CYP) enzymes to several classes of oxygenated metabolites with
potent biological activities. Major metabolites include
20-hydroxyeicosatetraenoic acid (20-HETE) and a series of regio-
and stereoisomeric epoxyeicosatrienoic acids (EETs). CYP4A and
CYP4F isoforms produce 20-HETE and CYP2C and CYP2J isoforms
EETs.
[0012] It is known that EPA (20:5, n-3) and DHA (22:6, n-3) may
serve as alternative substrates for AA-metabolizing CYP isoforms
(Arnold C. et al., J Biol Chem. 2010 Oct. 22; 285(43):32720-33.;
Fischer R. et al., J Lipid Res. 2014 Mar. 16; 55(6):1150-1164.).
CYP2C and CYP2J subfamily members that epoxidize AA to EETs,
metabolize EPA to epoxyeicosatetraenoic acids (EEQs), and DHA to
epoxydocosapentaenoic acids (EDPs). The .omega.-3 double bond
distinguishing EPA and DHA from AA is the preferred site of attack
by most of the epoxygenases resulting in the formation of 17,18-EEQ
and 19,20-EDP as main metabolites. CYP4A and CYP4F isoforms,
hydroxylating AA to 20-HETE, metabolize EPA to
20-hydroxyeicosapentaenoic acid (20-HEPE) and DHA to
22-hydroxydocosahexaenoic acid (22-HDHA). CYP1A1, CYP2E1 and other
isoforms converting AA predominantly to 19-HETE show pronounced
.omega.-3 epoxygenase activities with EPA and DHA. Human CYP1A1
variants lead to differential eicosapentaenoic acid metabolite
patterns. Cytochrome P450-dependent eicosapentaenoic acid
metabolites are novel BK channel activators. A remarkable feature
of CYP-dependent n-3 PUFA metabolism is the preferred epoxidation
of the n-3 double bond, which distinguishes EPA and DHA from AA.
The resulting metabolites--17,18-EEQ from EPA and 19,20-EDP from
DHA--are unique in having no homolog within the series of AA
products. In line with the substrate specificity of the CYP
isoforms, dietary EPA/DHA supplementation causes a profound shift
from AA- to EPA- and DHA-derived epoxy- and
.omega.-hydroxy-metabolites in all major organs and tissues of the
rat and presumably also in human.
[0013] EETs and 20-HETE play important roles in the regulation of
various cardiovascular functions (Roman R J., Physiol Rev. 2002;
82:131-85). It has been shown that Ang II-induced hypertension is
associated with a down-regulation of CYP-dependent AA metabolism
(Kaergel et I., Hypertension. 2002; 40:273-9) in a
double-transgenic rat (dTGR) model of Ang II-induced hypertension
and end-organ damage (Luft et al., Hypertension. 1999; 33:212-8).
The transgenic rats harbor the human renin and angiotensinogen
genes, produce Ang II locally and develop significant hypertension,
myocardial infarction and albuminuria. The animals die of
myocardial and renal failure before the eighth week of age. The
model shows severe features of Ang II-induced inflammation.
Reactive oxygen species are generated, the transcription factors
NF-.kappa.B and AP-1 are activated, and genes harboring binding
sites for these transcription factors are activated.
[0014] Recently, it has been shown that eicosapentaenoic acid (EPA)
supplementation significantly reduced the mortality of dTGR (Theuer
et al., Kidney Int. 2005; 67:248-58). Additionally, it has been
shown that dTGR develop ventricular arrhythmias based on Ang
II-induced electrical remodeling (Fischer et sl. Am J Physiol Heart
Circ Physiol. 2007; 293:H1242-1253). Treatment of the dTGR rats
with a PPAR-alpha activator strongly induced CYP2C23-dependent EET
production and protected against hypertension and end-organ damage
(Muller et al., Am J Pathol. 2004; 164:521-32).
[0015] Long-term feeding of dTGR (from week 4 to 7 of age) with a
mixture of pure EPA- and DHA-ethyl esters (Omacor from Solvay
Arzneimittel, Hannover, Germany) improved the electrical remodeling
of the heart in this model of angiotensin II-induced hypertension.
In particular, EPA and DHA reduced the mortality, suppressed the
inducibility of cardiac arrhythmias and protected against connexin
43-gap junctional remodeling (Fischer et al., Hypertension. 2008
February; 51(2):540-6). In general, CYP-dependent eicosanoids have
to be considered as second messengers: EETs and 20-HETE are
produced by CYP enzymes after extracellular signal induced release
of AA from membrane phospholipids (by phospholipase A2) and exert
their function in the context of signaling pathways modulating ion
transport, cell proliferation and inflammation. Depending on the
diet, n-3 PUFAs partially replace AA at the sn2-position of
phospholipids and may thus become involved as alternative molecules
in the subsequent signaling pathways.
[0016] The few studies on the biological activities of
CYP-dependent eicosanoids in the heart indicate important roles for
EETs and 20-HETE in the regulation of L-type Ca.sup.2+ and
sarcolemmal and mitochondrial ATP-sensitive potassium (KATP)
channels. In cardiac myocytes, L-type Ca.sup.2+ currents and cell
shorting are reduced upon inhibition of EET generation and these
effects can be reversed by adding 11,12-EET (Xiao et al., J
Physiol. 1998; 508 (Pt 3):777-92). EETs were also shown to activate
cardiac K.sub.ATP channels. This effect is highly stereoselective:
only the S,R but not the R,S-enantiomer of 11,12-EET was effective
(Lu et al., Mol Pharmacol. 2002; 62:1076-83). Overexpression of the
EET-generating human CYP2J2 resulted in an improved postischemic
functional recovery of the transgenic mouse heart via activation of
K.sub.ATP channels (Seubert et al., Circ Res. 2004; 95:506-14).
20-HETE appears to play an opposite role by acting as an endogenous
KATP channel blocker (Gross et al., J Mol Cell Cardiol. 2004;
37:1245-9; Nithipatikom et al., Circ Res. 2004; 95:e65-71).
[0017] Although n-3 PUFA-derived CYP metabolites, such as 17,18-EEQ
and 19,20-EDP, play important roles in mediating the beneficial
effects of n-3 PUFAs in the mammalian body, they are not used as
therapeutics due to their limited bioavailability as well as
chemical and metabolic instability. These epoxymetabolites of n-3
PUFAs are prone to autoxidation, rapid inactivation by the soluble
epoxide hydrolase, and degradation by .beta.-oxidation.
[0018] Therefore, the problem underlying the present invention is
to provide improved analogues of n-3 PUFA metabolites for treating
or reducing the risk of developing or preventing disorders
associated with neovascularization and/or inflammation, in
particular ophthalmic disorders associated with neovascularization
and/or inflammation,.
[0019] In a first aspect the above problem is solved by the
provision of compounds of the general formula (I):
P-E-I (I)
or a pharmaceutically acceptable salt thereof, wherein [0020] P is
a group represented by the general formula (II):
[0020] --(CH.sub.2).sub.n--O--(CH.sub.2).sub.k--X (II)
wherein [0021] n is 0 or an integer of from 3 to 8, i.e. 3, 4, 5,
6, 7, or 8, preferably 3; and [0022] k is 0, 1, or 2; preferably
with the proviso that when n is 0 k is 1, most preferably k is 1;
[0023] X represents CH.sub.2OH, CH.sub.2OAc, CH(O) or a group
selected from the group consisting of:
##STR00001##
[0023] preferably X is
##STR00002##
wherein [0024] 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);
[0025] R.sup.1 represents a hydroxyl group, C.sub.1-C.sub.6alkoxy,
--NHCN, --NH(C.sub.1-C.sub.6alkyl),
--NH(C.sub.3-C.sub.6cycloalkyl), --NH(aryl), or
--O(C.sub.1-C.sub.6alkyldiyl)O(C.dbd.O)R.sup.11; R.sup.11 is a
C.sub.1-C.sub.6alkyl group which is optionally substituted with one
or more fluorine or chlorine atom(s); or 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);
[0026] R.sup.2 represents --NHR.sup.3; --NR.sup.20R.sup.21;
--OR.sup.22; --(OCH.sub.2--CH.sub.2).sub.i--R.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; [0027] or is selected from the group consisting of:
##STR00003##
[0027] wherein [0028] 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(.dbd.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(.dbd.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(.dbd.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(.dbd.O)OR.sup.51; [0029]
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; [0030] 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);
[0031] 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); [0032] 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.1O-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); [0033]
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; [0034] R.sup.23 is --OH,
--O(C.sub.1-C.sub.3)alkyl, or --N(C.sub.1-C.sub.3)dialkyl; [0035] i
is an integer of from 1 to 10; [0036] R.sup.24, R.sup.25, and
R.sup.26 each independently represents a hydrogen atom;
--C(.dbd.O)C.sub.11-C.sub.21alkyl; or
--C(.dbd.O)C.sub.11-C.sub.21alkenyl; [0037] 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
[0037] ##STR00004## [0038] 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(.dbd.O) by an amide
bond; [0039] o is an integer of from 1 to 10; [0040] R.sup.4 is
selected from the group consisting of:
[0040] ##STR00005## [0041] h is 0, 1, or 2; [0042] R.sup.5
represents a hydrogen atom; a fluorine or chlorine atom;
--CF.sub.3; --C(.dbd.O)OR.sup.51; [0043] --NHC(.dbd.O)R.sup.52;
--C(.dbd.O)NR.sup.53R.sup.54; or --S(O.sub.2)OH; [0044] 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; [0045] 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; [0046] 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(C.dbd.O)(C.sub.1-C.sub.6)alkyl
group, or a
--O(C.sub.1-C.sub.6)alkyldiylO(C.dbd.O)(C.sub.2-C.sub.6)alkenyl
group; wherein the C.sub.1-C.sub.6alkyl group and the
C.sub.2-C.sub.6alkenyl group 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 [0047] R.sup.6
represents a hydroxyl group and R.sup.7 represents a group:
[0047] ##STR00006## [0048] 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(.dbd.O)-aryl, --C(.dbd.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), [0049]
--N(C.sub.1-C.sub.6)dialkyl, and an oxo substituent; [0050] g is 1
or 2, preferably 2; [0051] X.sup.1 represents an oxygen atom;
sulfur atom; or NH; [0052] X.sup.2 represents an oxygen atom;
sulfur atom; NH; or N(CH.sub.3); [0053] X.sup.3 represents an
oxygen atom; sulfur atom; nitrogen atom; carbon atom; or C--OH; and
the dashed line represents a carbon-carbon bond or a carbon-carbon
double bond; [0054] E is a group represented by the general formula
(III) or (IV):
[0054] ##STR00007## [0055] wherein R12 and R13 are preferably in
cis configuration, and wherein [0056] 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; [0057] 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(.dbd.O)-aryl, --C(.dbd.O)C.sub.1-C.sub.6alkyl, or
--SO.sub.2(C.sub.1-C.sub.6alkyl); or --SO.sub.2aryl; wherein any of
the foregoing C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy, or aryl
are optionally substituted with one, two or three substituents
independently selected from the group consisting of --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N(C.sub.1-C.sub.6)dialkyl,
C.sub.1-C.sub.6alkylcarbonyloxy-,
C.sub.1-C.sub.6alkoxycarbonyloxy-,
C.sub.1-C.sub.6alkylcarbonylthio-,
C.sub.1-C.sub.6alkylaminocarbonyl-,
di(C.sub.1-C.sub.6)alkylaminocarbonyl-, fluorine or chlorine atom,
and 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,
--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; [0058] l is --(CH.sub.2).sub.m--Y, wherein [0059] m
is an integer of from 3 to 6, i.e 3, 4, 5, or 6, provided that m is
an integer of from 3 to 5 when E is a group according to general
formula (III); [0060] Y represents
--U--V--W--(CH.sub.2).sub.p--(CH.sub.3).sub.q, wherein p is an
integer from 0 to 6; q is 0 or 1; U is absent or selected from the
group consisting of CH, CH.sub.2 and NR.sup.40, with the proviso
that U is only CH if it forms an epoxy group together with V and W;
V is selected from the group consisting of --C(O)--,
--C(O)--C(O)--, --O--, and --S--; W is selected from the group
consisting of CH, CH.sub.2 and NR.sup.40 with the proviso that W is
only CH if it forms an epoxy group together with U and V; [0061] or
Y represents a group selected from the group consisting of:
##STR00008##
[0061] wherein [0062] 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(.dbd.O)aryl, or
--C(.dbd.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; [0063] 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)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; [0064] f is an integer of from 0 to 2; [0065] with
the proviso that [0066] when X does not comprise a
--C(.dbd.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 [0067] for use in treating, reducing the risk of
developing or preventing a disorder associated with
neovascularization and/or inflammation. [0068] In a preferred
embodiment, the compounds of present invention are compounds of
formula (I) as described above with the proviso that [0069] when X
does not comprise a --C(.dbd.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. [0070] In a preferred embodiment, the
compounds of formula (I) are compounds as described above with the
further proviso that [0071] when n is 3, 5, 6, 7 or 8, preferably 3
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; [0072] P represents a group:
[0072] --(CH.sub.2).sub.3--O--(CH.sub.2)--X.sup.81;
--(CH.sub.2).sub.5--O--(CH.sub.2)--X.sup.81;
wherein [0073] X.sup.81 represents a group selected from the group
consisting of:
##STR00009##
[0074] R.sup.10 is defined as R.sup.1 above; [0075] R.sup.2'
represents --NHR.sup.3'; --OR.sup.22';
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.O) by an ester
bond via the 1-O-, 3-O-, or 6-O-position of the saccharide; [0076]
or wherein R.sup.2 is selected from the group consisting of:
##STR00010##
[0076] wherein [0077] 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); [0078] 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,
--NH(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; [0079] R.sup.23 and i are as defined above;
[0080] R.sup.24, R.sup.25, R.sup.26, and R.sup.27 are as defined
above; [0081] R.sup.4' is defined as R.sup.4 above; and h is
defined as above; [0082] R.sup.6' and R.sup.7' are defined as
R.sup.6 and R.sup.7 above; [0083] R.sup.8'' and R.sup.8'' are
defined as R.sup.8 and R.sup.8' above; [0084] 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.
[0085] In a more preferred embodiment the compound of the present
invention is one, wherein X is
##STR00011## [0086] wherein R.sup.2 is --OR.sup.22;
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.O) by an ester
bond via the 1-O-, 3-O-, or 6-O-position of the saccharide; [0087]
or wherein R.sup.2 is selected from the group consisting of:
[0087] ##STR00012## [0088] wherein R.sup.23 and i are as defined
above, preferable i is 3; [0089] and wherein R.sup.22, and R.sup.23
to R.sup.27 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.
[0090] In a further more preferred embodiment, the compound of the
present invention is one, wherein X is --C(.dbd.O)OH or a suitable
salt of the carboxylic acid, preferably a free carboxylic acid.
[0091] In another more preferred embodiment, the compound of the
present invention is one, wherein Y is one of the oxamides as
defined above.
[0092] It is further preferred that the compound of the present
invention is one, wherein X is
##STR00013## [0093] wherein R.sup.2 is --OR.sup.22;
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.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:
[0093] ##STR00014## [0094] wherein and R.sup.22, R.sup.23 to
R.sup.27 and i are as defined above, preferably R.sup.22 is a
hydrogen atom or a C.sub.1-C.sub.6alkyl group, more preferably a
hydrogen atom, preferably i is 2 to 4, more preferably 3, and
wherein Y is preferably one of the oxamides defined above.
[0095] In a more preferred embodiment, the compound of the present
invention is one, wherein X is C(.dbd.O)OH, preferably the free
carboxylic acid, and Y is preferably one of the oxamides defined
above.
[0096] In another more preferred embodiment, the compound of the
present invention is one with the following formula (V):
##STR00015##
wherein [0097] R.sup.55 represents --OH; --OR.sup.22;
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; a mono-, or disaccharide,
or a derivative thereof, which is joined to --C(.dbd.O) by an ester
bond via the 1-O-, 3-O-, or 6-O-position of the saccharide; [0098]
R.sup.22, R.sup.23 and i are as defined above, 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;
[0099] Y represents a group selected from the group consisting
of:
##STR00016##
[0099] wherein
##STR00017##
are preferred, and
##STR00018##
is particularly preferred; and [0100] wherein R.sup.40 to R.sup.50
are defined above, preferably R.sup.40 is a hydrogen atom or a
C.sub.1-C.sub.6alkyl group, more preferably a hydrogen atom [0101]
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; [0102] s is 0,
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; [0103] 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. [0104] In a further most preferred
embodiment the compounds of formula (V) are those wherein [0105]
R.sup.55 represents --OH or
--(OCH.sub.2--CH.sub.2).sub.i--R.sup.23; i is 2 to 4, preferably i
is 3; R.sup.23 is preferably OH; [0106] Y is an oxamide, a
carbamide or a carbamate, preferably a C.sub.1-C.sub.6alkyl
substituted oxamide, carbamide or carbamate; [0107] 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 [0108] 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.
[0109] The most preferred specific compounds of the present
invention are those selected from the group consisting of:
##STR00019## ##STR00020## ##STR00021## ##STR00022##
and [0110] or a pharmaceutically acceptable salt thereof.
[0111] Among the above, the compound with the following formula
(VI)
##STR00023##
or a pharmaceutically acceptable salt thereof is most
preferred.
[0112] The compounds of the present invention have the advantage as
demonstrated below in the experimental section that they are
effective for treating, reducing the risk of developing or
preventing a disorder associated with neovascularization and/or
inflammation, in particular an ophthalmic disorder associated with
neovascularization and/or inflammation. They are at the same time
metabolically robust for pharmaceutical formulation and
administration to subjects in need thereof.
[0113] The compounds described herein are generally described using
standard nomenclature. For compounds having asymmetric centers, it
is understood that, unless otherwise specified, all of the optical
isomers and mixtures thereof are encompassed. Compounds with two or
more asymmetric elements can also be present as mixtures of
diastereomers. In addition, compounds with carbon-carbon double
bonds may occur in Z- and E-forms, with all isomeric forms of the
compounds being included in the present invention unless otherwise
specified. Where a compound exists in various tautomeric forms, a
recited compound is not limited to any one specific tautomer, but
rather is intended to encompass all tautomeric forms. Recited
compounds are further intended to encompass compounds in which one
or more atoms are replaced with an isotope, i.e., an atom having
the same atomic number but a different mass number. By way of
general example, and without limitation, isotopes of hydrogen
include tritium and deuterium and isotopes of carbon include
.sup.11C, .sup.13C, and .sup.14C.
[0114] Compounds according to the formulas provided herein, which
have one or more stereogenic center(s), have an enantiomeric excess
of at least 50%. For example, such compounds may have an
enantiomeric excess of at least 60%, 70%, 80%, 85%, 90%, 95%, or
98%. Some embodiments of the compounds have an enantiomeric excess
of at least 99%. It will be apparent that single enantiomers
(optically active forms) can be obtained by asymmetric synthesis,
synthesis from optically pure precursors, biosynthesis, e.g. using
modified CYP102 (CYP BM-3) or by resolution of the racemates, e.g.
enzymatic resolution or resolution by conventional methods such as
crystallization in the presence of a resolving agent, or
chromatography, using, for example, a chiral HPLC column.
[0115] Certain compounds are described herein using a general
formula that includes variables such as, e.g. P, E, I,
R.sup.1-R.sup.50, X--X.sup.81, and Y. Unless otherwise specified,
each variable within such a formula is defined independently of any
other variable, and any variable that occurs more than one time in
a formula is defined independently at each occurrence. Thus, for
example, if a group is shown to be substituted with 0-2 R*, the
group may be unsubstituted or substituted with up to two R* groups,
and R* at each occurrence is selected independently from the
definition of R*. Also, combinations of substituents and/or
variables are permissible only if such combinations result in
stable compounds, i.e., compounds that can be isolated,
characterized and tested for biological activity.
[0116] A "pharmaceutically acceptable salt" of a compound disclosed
herein is an acid or base salt that is generally considered in the
art to be suitable for use in contact with the tissues of human
beings or animals without excessive toxicity or carcinogenicity,
and preferably without irritation, allergic response, or other
problem or complication. Such salts include mineral and organic
acid salts of basic residues such as amines, as well as alkali or
organic salts of acidic residues such as carboxylic acids.
[0117] Suitable pharmaceutical salts include, but are not limited
to, salts of acids such as hydrochloric, phosphoric, hydrobromic,
malic, glycolic, fumaric, sulfuric, sulfamic, sulfanilic, formic,
toluenesulfonic, methanesulfonic, benzenesulfonic, ethane
disulfonic, 2-hydroxyethylsulfonic, nitric, benzoic,
2-acetoxybenzoic, citric, tartaric, lactic, stearic, salicylic,
glutamic, ascorbic, pamoic, succinic, fumaric, maleic, propionic,
hydroxymaleic, hydroiodic, phenylacetic, alkanoic such as acetic,
HOOC--(CH.sub.2).sub.n--COOH where n is any integer from 0 to 6,
i.e. 0, 1, 2, 3, 4, 5 or 6, and the like. Similarly,
pharmaceutically acceptable cations include, but are not limited to
sodium, potassium, calcium, aluminum, lithium and ammonium. Those
of ordinary skill in the art will recognize further
pharmaceutically acceptable salts for the compounds provided
herein. In general, a pharmaceutically acceptable acid or base salt
can be synthesized from a parent compound that contains a basic or
acidic moiety by any conventional chemical method. Briefly, such
salts can be prepared by reacting the free acid or base forms of
these compounds with a stoichiometric amount of the appropriate
base or acid in water or in an organic solvent, or in a mixture of
the two. Generally, the use of nonaqueous media, such as ether,
ethyl acetate, ethanol, isopropanol or acetonitrile, is
preferred.
[0118] It will be apparent that each compound of formula (I) may,
but need not, be present as a hydrate, solvate or non-covalent
complex. In addition, the various crystal forms and polymorphs are
within the scope of the present invention as are prodrugs of the
compounds of formula (I) provided herein.
[0119] A "prodrug" is a compound that may not fully satisfy the
structural requirements of the compounds provided herein, but is
modified in vivo, following administration to a subject or patient,
to produce a compound of formula (I) provided herein. For example,
a prodrug may be an acylated derivative of a compound as provided
herein. Prodrugs include compounds wherein hydroxy, carboxy, amine
or sulfhydryl groups are bonded to any group that, when
administered to a mammalian subject, cleaves to form a free
hydroxy, carboxy, amino, or sulfhydryl group, respectively.
Examples of prodrugs include, but are not limited to, acetate,
formate, phosphate and benzoate derivatives of alcohol and amine
functional groups within the compounds provided herein. Prodrugs of
the compounds provided herein may be prepared by modifying
functional groups present in the compounds in such a way that the
modifications are cleaved in vivo to generate the parent
compounds.
[0120] A "substituent," as used herein, refers to a molecular
moiety that is covalently bonded to an atom within a molecule of
interest. For example, a "ring substituent" may be a moiety such as
a halogen, alkyl group, haloalkyl group or other substituent
described herein that is covalently bonded to an atom, preferably a
carbon or nitrogen atom, that is a ring member. The term
"substituted," as used herein, means that any one or more hydrogens
on the designated atom is replaced with a selection from the
indicated substituents, provided that the designated atom's normal
valence is not exceeded, and that the substitution results in a
stable compound, i.e., a compound that can be isolated,
characterized and tested for biological activity. When a
substituent is oxo, i.e., .dbd.O, then 2 hydrogens on the atom are
replaced. An oxo group that is a substituent of an aromatic carbon
atom results in a conversion of --CH-- to --C(.dbd.O)-- and a loss
of aromaticity. For example a pyridyl group substituted by oxo is a
pyridone.
[0121] The expression "optionally substituted" refers to a group in
which one, two, three or more hydrogen atoms may have been replaced
independently of each other by the respective substituents.
[0122] As used herein, the term "amino acid" refers to any organic
acid containing one or more amino substituents, e.g. .alpha.-,
.beta.- or .gamma.-amino, derivatives of aliphatic carboxylic
acids. In the polypeptide notation used herein, e.g. Xaa.sub.5,
i.e. Xaa.sub.1Xaa.sub.2Xaa.sub.3Xaa.sub.4Xaa.sub.5, wherein
Xaa.sub.1 to Xaa.sub.5 are each and independently selected from
amino acids as defined, the left hand direction is the amino
terminal direction and the right hand direction is the carboxy
terminal direction, in accordance with standard usage and
convention.
[0123] The term "conventional amino acid" refers to the twenty
naturally occurring amino acids, and encompasses all stereomeric
isoforms, i.e. D,L-, D- and L-amino acids thereof. These
conventional amino acids can herein also be referred to by their
conventional three-letter or one-letter abbreviations and their
abbreviations follow conventional usage (see, for example,
Immunology--A Synthesis, 2.sup.nd Edition, E. S. Golub and D. R.
Gren, Eds., Sinauer Associates, Sunderland Mass. (1991)).
[0124] The term "non-conventional amino acid" refers to unnatural
amino acids or chemical amino acid analogues, e.g.
.alpha.,.alpha.-disubstituted amino acids, N-alkyl amino acids,
homo-amino acids, dehydroamino acids, aromatic amino acids (other
than phenylalanine, tyrosine and tryptophan), and ortho-, meta- or
para-aminobenzoic acid. Non-conventional amino acids also include
compounds which have an amine and carboxyl functional group
separated in a 1,3 or larger substitution pattern, such as
.beta.-alanine, .gamma.-amino butyric acid, Freidinger lactam, the
bicyclic dipeptide (BTD), amino-methyl benzoic acid and others well
known in the art. Statine-like isosteres, hydroxyethylene
isosteres, reduced amide bond isosteres, thioamide isosteres, urea
isosteres, carbamate isosteres, thioether isosteres, vinyl
isosteres and other amide bond isosteres known to the art may also
be used. The use of analogues or non-conventional amino acids may
improve the stability and biological half-life of the added peptide
since they are more resistant to breakdown under physiological
conditions. The person skilled in the art will be aware of similar
types of substitution which may be made. A non limiting list of
non-conventional amino acids which may be used as suitable building
blocks for a peptide and their standard abbreviations (in brackets)
is as follows: .alpha.-aminobutyric acid (Abu), L-N-methylalanine
(Nmala), .alpha.-amino-.alpha.-methylbutyrate (Mgabu),
L-N-methylarginine (Nmarg), aminocyclopropane (Cpro),
L-N-methylasparagine (Nmasn), carboxylate L-N-methylaspartic acid
(Nmasp), aniinoisobutyric acid (Aib), L-N-methylcysteine (Nmcys),
aminonorbornyl (Norb), L-N-methylglutamine (Nmgln), carboxylate
L-N-methylglutamic acid (Nmglu), cyclohexylalanine (Chexa),
L-N-methylhistidine (Nmhis), cyclopentylalanine (Cpen),
L-N-methylisolleucine (Nmile), L-N-methylleucine (Nmleu),
L-N-methyllysine (Nmlys), L-N-methylmethionine (Nmmet),
L-N-methylnorleucine (Nmnle), L-N-methylnorvaline (Nmnva),
L-N-methylornithine (Nmorn), L-N-methylphenylalanine (Nmphe),
L-N-methylproline (Nmpro), L-N-methylserine (Nmser),
L-N-methylthreonine (Nmthr), L-N-methyltryptophan (Nmtrp),
D-ornithine (Dorn), L-N-methyltyrosine (Nmtyr), L-N-methylvaline
(Nmval), L-N-methylethylglycine (Nmetg), L-N-methyl-t-butylglycine
(Nmtbug), L-norleucine (Nle), L-norvaline (Nva),
.alpha.-methyl-aminoisobutyrate (Maib),
.alpha.-methyl-.gamma.-aminobutyrate (Mgabu),
D-.alpha.-methylalanine (Dmala), .alpha.-methylcyclohexylalanine
(Mchexa), D-.alpha.-methylarginine (Dmarg),
.alpha.-methylcylcopentylalanine (Mcpen),
D-.alpha.-methylasparagine (Dmasn),
.alpha.-methyl-.alpha.-napthylalanine (Manap),
D-.alpha.-methylaspartate (Dmasp), .alpha.-methylpenicillamine
(Mpen), D-.alpha.-methylcysteine (Dmcys), N-(4-aminobutyl)glycine
(Nglu), D-.alpha.-methylglutamine (Dmgln), N-(2-aminoethyl)glycine
(Naeg), D-.alpha.-methylhistidine (Dmhis), N-(3-aminopropyl)glycine
(Norn), D-.alpha.-methylisoleucine (Dmile),
N-amino-.alpha.-methylbutyrate (Nmaabu), D-.alpha.-methylleucine
(Dmleu), .alpha.-napthylalanine (Anap), D-.alpha.-methyllysine
(Dmlys), N-benzylglycine (Nphe), D-.alpha.-methylmethionine
(Dmmet), N-(2-carbamylethyl)glycine (Ngln),
D-.alpha.-methylornithine (Dmorn), N-(carbamyl-methyl)glycine
(Nasn), D-.alpha.-methylphenylalanine (Dmphe),
N-(2-carboxyethyl)glycine (Nglu), D-.alpha.-methylproline (Dmpro),
N-(carboxymethyl)glycine (Nasp), D-.alpha.-methylserine (Dmser),
N-cyclobutylglycine (Ncbut), D-.alpha.-methylthreonine (Dmthr),
N-cycloheptylglycine (Nchep), D-.alpha.-methyltryptophan (Dmtrp),
N-cyclohexylglycine (Nchex), D-.alpha.-methyltyrosine (Dmty),
N-cyclo-decylglycine (Ncdec), D-.alpha.-methylvaline (Dmval),
N-cylcododecylglycine (Ncdod), D-N-methylalanine (Dnmala),
N-cyclooctylglycine (Ncoct), D-N-methylarginine (Dnmarg),
N-cyclopropylglycine (Ncpro), D-N-methylasparagine (Dnmasn),
N-cycloundecylglycine (Ncund), D-N-methylaspartate (Dnmasp),
N-(2,2-diphenylethyl)glycine (Nbhm), D-N-methylcysteine (Dnmcys),
N-(3,3-diphenylpropyl)glycine (Nbhe), D-N-methylglutamine (Dnmgln),
N-(3-guanidinopropyl)glycine (Narg), D-N-methylglutamate (Dnmglu),
N-(1-hydroxyethyl)glycine (Ntbx), D-N-methylhistidine (Dnmhis),
N-(hydroxyethyl))glycine (Nser), D-N-methylisoleucine (Dnmile),
N-(imidazolylethyl))glycine (Nhis), D-N-methylleucine (Dnmleu),
N-(3-indolylyethyl)glycine (Nhtrp), D-N-methyllysine (Dnnilys),
N-methyl-.gamma.-aminobutyrate (Nmgabu), N-methylcyclohexylalanine
(Nmchexa), D-N-methylmethionine (Dnmmet), D-N-methylornithine
(Dnmorn), N-methylcyclopentylalanine (Nmcpen), N-methylglycine
(Nala), D-N-methylphenylalanine (Dnmphe), N-methylaminoisobutyrate
(Nmaib), D-N-methylproline (Dnmpro), N-(1-methylpropyl)glycine
(Nile), D-N-methylserine (Dnmser), N-(2-methylpropyl)glycine
(Nleu), D-N-methylthreonine (Dnmthr), D-N-methyltryptophan
(Dnmtrp), N-(1-methylethyl)glycine (Nval), D-N-methyltyrosine
(Dnmtyr), N-methyla-napthylala nine (Nmanap), D-N-methylvaline
(Dnmval), N-methylpenicillamine (Nmpen), .gamma.-aminobutyric acid
(Gabu), N-(p-hydroxyphenyl)glycine (Nhtyr), L-/- butylglycine
(Tbug), N-(thiomethyl)glycine (Ncys), L-ethylglycine (Etg),
penicillamine (Pen), L-homophenylalanine (Hphe),
L-.alpha.-methylalanine (Mala), L-.alpha.-methylarginine (Marg),
L-.alpha.-methylasparagine (Masn), L-.alpha.-methylaspartate
(Masp), L-.alpha.-methyl-t-butylglycine (Mtbug),
L-.alpha.-methylcysteine (Mcys), L-methylethylglycine (Metg),
L-.alpha.-methylglutamine (Mgln), L-.alpha.-methylglutamate (Mglu),
L-.alpha.-methylhistidine (Mhis), L-.alpha.-methylhomophenylalanine
(Mhphe), L-.alpha.-methylisoleucine (Mile),
N-(2-methylthioethyl)glycine (Nmet), L-.alpha.-methylleucine
(Mleu), L-.alpha.-methyllysine (Mlys), L-.alpha.-methylmethionine
(Mmet), L-.alpha.-methylnorleucine (Mnle),
L-.alpha.-methylnorvaline (Mnva), L-.alpha.-methylornithine (Morn),
L-.alpha.-methylphenylalanine (Mphe), L-.alpha.-methylproline
(Mpro), L-.alpha.-methylserine (Mser), L-.alpha.-methylthreonine
(Mthr), L-.alpha.-methyltryptophan (Mtrp), L-.alpha.-methyltyrosine
(Mtyr), L-.alpha.-methylvaline (Mval), L-N-methylhomophenylalanine
(Nmhphe), N-(N-(2,2-diphenylethyl)carbamylmethyl)glycine (Nnbhm),
N-(N-(3,3-diphenylpropyl)-carbamylmethyl)glycine (Nnbhe),
1-carboxy-1-(2,2-diphenyl-ethylamino)cyclopropane (Nmbc),
L-O-methyl serine (Omser), L-O-methyl homoserine (Omhser).
[0125] The expression alkyl refers to a saturated, straight-chain
or branched hydrocarbon group that contains from 1 to 20 carbon
atoms, preferably from 1 to 10 carbon atoms, e.g. a n-octyl group,
especially from 1 to 6, i.e. 1, 2, 3, 4, 5, or 6, carbon atoms, for
example a methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl,
sec-butyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl, or
2,2-dimethylbutyl.
[0126] The expression alkenyl refers to an at least partially
unsaturated, straight-chain or branched, hydrocarbon group that
contains from 2 to 21 carbon atoms, preferably from 2 to 6 carbon
atoms, i.e. 2, 3, 4, 5 or 6 carbon atoms, for example an ethenyl
(vinyl), propenyl (allyl), iso-propenyl, butenyl, isoprenyl or
hex-2-enyl group, or from 11 to 21 carbon atoms, i.e. 11, 12, 13,
14, 15, 16, 17, 18, 19, 20 or 21 carbon atoms, for example a
hydrocarbon group comprising a methylene chain interrupted by one
double bond as, for example, found in monounsaturated fatty acids
or a hydrocarbon group comprising methylene-interrupted polyenes,
e.g. hydrocarbon groups comprising two or more of the following
structural unit --[CH.dbd.CH--CH.sub.2]--, as, for example, found
in polyunsaturated fatty acids. Alkenyl groups have one or more,
preferably 1, 2, 3, 4, 5, or 6 double bond(s).
[0127] The expression alkynyl refer to at least partially
unsaturated, straight-chain or branched hydrocarbon groups that
contain from 2 to 20 carbon atoms, preferably from 2 to 10 carbon
atoms, especially from 2 to 6, i.e. 2, 3, 4, 5 or 6, carbon atoms,
for example an ethinyl, propinyl, butinyl, acetylenyl, or propargyl
group. Preferably, alkynyl groups have one or two (especially
preferably one) triple bond(s).
[0128] Furthermore, the terms alkyl, alkenyl and alkynyl refer to
groups in which one or more hydrogen atom(s) have been replaced,
e.g. by a halogen atom, preferably F or Cl, such as, for example, a
2,2,2-trichloroethyl or a trifluoromethyl group.
[0129] The expression heteroalkyl refers to an alkyl, alkenyl or
alkynyl group in which one or more, preferably 1, 2 or 3, carbon
atoms, have been replaced independently of each other by an oxygen,
nitrogen, phosphorus, boron, selenium, silicon or sulfur atom,
preferably by an oxygen, sulfur or nitrogen atom. The expression
heteroalkyl can also refer to a carboxylic acid or to a group
derived from a carboxylic acid, such as, for example, acyl,
acylalkyl, alkoxycarbonyl, acyloxy, acyloxyalkyl, carboxyalkylamide
or alkoxycarbonyloxy.
[0130] Preferably, a heteroalkyl group contains from 1 to 10 carbon
atoms and from 1 to 4 hetero atoms selected from oxygen, nitrogen
and sulphur (especially oxygen and nitrogen). Especially
preferably, a heteroalkyl group contains from 1 to 6, i.e. 1, 2, 3,
4, 5, or 6, carbon atoms and 1, 2 or 3, especially 1 or 2, hetero
atoms selected from oxygen, nitrogen and sulphur, especially oxygen
and nitrogen.
[0131] Examples of heteroalkyl groups are groups of formulae:
R.sup.a--O--Y.sup.a--, R.sup.a--S--Y.sup.a--,
R.sup.a--N(R.sup.b)--Y.sup.a--, R.sup.a--CO--Y.sup.a--,
R.sup.a--O--CO--Y.sup.a--, R.sup.a--CO--O--Y.sup.a--,
R.sup.a--CO--N(R.sup.b)--Y.sup.a--,
R.sup.a--N(R.sup.b)--CO--Y.sup.a--,
R.sup.a--O--CO--N(R.sup.b)--Y.sup.a--,
R.sup.a--N(R.sup.b)--CO--O--Y.sup.a--,
R.sup.a--N(R.sup.b)--CO--N(R.sup.c)--Y.sup.a--,
R.sup.a--O--CO--O--Y.sup.a--,
R.sup.a--N(R.sup.b)--C(.dbd.NR.sup.d)--N(R.sup.c)--Y.sup.a--,
R.sup.a--CS--Y.sup.a--, R.sup.a--O--CS--Y.sup.a--,
R.sup.a--CS--O--Y.sup.a--, R.sup.a--CS--N(R.sup.b)--Y.sup.a--,
R.sup.a--N(R.sup.b)--CS--Y.sup.a--,
R.sup.a--O--CS--N(R.sup.b)--Y.sup.a--,
R.sup.a--N(R.sup.b)--CS--O--Y.sup.a--,
R.sup.a--N(R.sup.b)--CS--N(R.sup.c)--Y.sup.a--,
R.sup.a--O--CS--O--Y.sup.a--, R.sup.a--S--CO--Y.sup.a--,
R.sup.a--CO--S--Y.sup.a--, R.sup.a--S--CO--N(R.sup.b)--Y.sup.a--,
R.sup.a--N(R.sup.b)--CO--S--Y.sup.a--,
R.sup.a--S--CO--O--Y.sup.a--, R.sup.a--O--CO--S--Y.sup.a--,
R.sup.a--S--CO--S--Y.sup.a--, R.sup.a--S--CS--Y.sup.a--,
R.sup.a--CS--S--Y.sup.a--, R.sup.a--S--CS--N(R.sup.b)--Y.sup.a--,
R.sup.a--N(R.sup.b)--CS--S--Y.sup.a--,
R.sup.a--S--CS--O--Y.sup.a--, R.sup.a--O--CS--S--Y.sup.a--, wherein
R.sup.a being a hydrogen atom, a C.sub.1-C.sub.6 alkyl, a
C.sub.2-C.sub.6 alkenyl or a C.sub.2-C.sub.6 alkynyl group; R.sup.b
being a hydrogen atom, a C.sub.1-C.sub.6 alkyl, a C.sub.2-C.sub.6
alkenyl or a C.sub.2-C.sub.6 alkynyl group; R.sup.c being a
hydrogen atom, a C.sub.1-C.sub.6 alkyl, a C.sub.2-C.sub.6 alkenyl
or a C.sub.2-C.sub.6 alkynyl group; R.sup.d being a hydrogen atom,
a C.sub.1-C.sub.6 alkyl, a C.sub.2-C.sub.6 alkenyl or a
C.sub.2-C.sub.6 alkynyl group and Y.sup.a being a direct bond, a
C.sub.1-C.sub.6alkylene, a C.sub.2-C.sub.6 alkenylene or a
C.sub.2-C.sub.6 alkynylene group, wherein each heteroalkyl group
contains at least one carbon atom and one or more hydrogen atoms
may be replaced by fluorine or chlorine atoms.
[0132] Specific examples of heteroalkyl groups are methoxy,
trifluoromethoxy, ethoxy, n-propyloxy, isopropyloxy, butoxy,
tert-butyloxy, methoxymethyl, ethoxymethyl, --CH.sub.2CH.sub.2OH,
--CH.sub.2OH, methoxyethyl, 1-methoxyethyl, 1-ethoxyethyl,
2-methoxyethyl or 2-ethoxyethyl, methylamino, ethylamino,
propylamino, isopropylamino, dimethylamino, diethylamino,
isopropylethylamino, methylamino methyl, ethylamino methyl,
diisopropylamino ethyl, methylthio, ethylthio, isopropylthio, enol
ether, dimethylamino methyl, dimethylamino ethyl, acetyl,
propionyl, butyryloxy, acetyloxy, methoxycarbonyl, ethoxycarbonyl,
propionyloxy, acetylamino or propionylamino, carboxymethyl,
carboxyethyl or carboxypropyl, N-ethyl-N-methylcarbamoyl or
N-methylcarbamoyl. Further examples of heteroalkyl groups are
nitrile, isonitrile, cyanate, thiocyanate, isocyanate,
isothiocyanate and alkylnitrile groups.
[0133] The expression alkoxy refers to an alkyl group singular
bonded to oxygen.
[0134] The expression alkylthio refers to an alkyl group singular
bonded to sulfur.
[0135] The expressions cycloalkyl and carbocyclic ring refer to a
saturated cyclic group of hydrocarbons that contains one or more
rings, preferably 1 or 2), and contains from 3 to 14 ring carbon
atoms, preferably from 3 to 10, especially 3, 4, 5, 6 or 7 ring
carbon atoms, e.g. a cyclopropyl, cyclobutyl, cyclopentyl,
spiro[4,5]decanyl, norbornyl, cyclohexyl, decalinyl,
bicyclo[4.3.0]nonyl, tetraline, or cyclopentylcyclohexyl group. The
expression cycloalkyl refers furthermore to groups in which one or
more hydrogen atoms have been replaced by fluorine, chlorine,
bromine or iodine atoms or by OH, .dbd.O, SH, NH.sub.2, .dbd.NH,
N.sub.3 or NO.sub.2 groups, thus, for example, cyclic ketones such
as, for example, cyclohexanone, 2-cyclohexenone or cyclopentanone.
Further specific examples of cycloalkyl groups are a cyclopropyl,
cyclobutyl, cyclopentyl, spiro[4,5]decanyl, norbornyl, cyclohexyl,
cyclopentenyl, cyclohexadienyl, decalinyl, bicyclo[4.3.0]nonyl,
tetraline, cyclopentylcyclohexyl, fluorocyclohexyl or
cyclohex-2-enyl group.
[0136] The expression aryl refers to an aromatic group that
contains one or more rings containing from 6 to 14 ring carbon
atoms, preferably from 6 to 10, especially 6, ring carbon
atoms.
[0137] The expression heteroaryl refers to an aromatic group that
contains one or more rings containing from 5 to 14 ring atoms,
preferably from 5 to 10, especially 5 or 6, ring atoms, and
contains one or more, preferably 1, 2, 3 or 4, oxygen, nitrogen,
phosphorus or sulfur ring atoms, preferably O, S or N. Examples are
pyridyl (e.g. 4-pyridyl), imidazolyl (e.g. 2-imidazolyl),
phenylpyrrolyl (e.g. 3-phenylpyrrolyl), thiazolyl, isothiazolyl,
1,2,3-triazolyl, 1,2,4-triazolyl, oxadiazolyl,thiadiazolyl,
indolyl, indazolyl, tetrazolyl, pyrazinyl, pyrimidinyl,
pyridazinyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl,
isoxazolyl, indazolyl, indolyl, benzimidazolyl, benzoxazolyl,
benzisoxazolyl, benzthiazolyl, pyridazinyl, quinolinyl,
isoquinolinyl, pyrrolyl, purinyl, carbazolyl, acridinyl, pyrimidyl,
2,3'-bifuryl, pyrazolyl (e.g. 3-pyrazolyl) and isoquinolinyl
groups. The expression heterocycloalkyl refers to a cycloalkyl
group as defined above in which one or more (preferably 1, 2 or 3)
ring carbon atoms, each independently, have been replaced by an
oxygen, nitrogen, silicon, selenium, phosphorus or sulfur atom
(preferably by an oxygen, sulfur or nitrogen atom). A
heterocycloalkyl group has preferably 1 or 2 ring(s) containing
from 3 to 10 (especially 3, 4, 5, 6 or 7) ring atoms (preferably
selected from C, O, N and S). The expression heterocycloalkyl
refers furthermore to groups in which one or more hydrogen atoms
have been replaced by fluorine, chlorine, bromine or iodine atoms
or by OH, .dbd.O, SH, .dbd.S, NH.sub.2, .dbd.NH, N.sub.3 or
NO.sub.2 groups. Examples are a piperidyl, prolinyl,
imidazolidinyl, piperazinyl, morpholinyl, urotropinyl,
pyrrolidinyl, tetrahydrothiophenyl, tetrahydropyranyl,
tetrahydrofuryl or 2-pyrazolinyl group and also lactames, lactones,
cyclic imides and cyclic anhydrides.
[0138] The expression alkylcycloalkyl refers to a group that
contains both cycloalkyl and also alkyl, alkenyl or alkynyl groups
in accordance with the above definitions, for example
alkylcycloalkyl, cycloalkylalkyl, alkylcycloalkenyl,
alkenylcycloalkyl and alkynylcycloalkyl groups. An alkylcycloalkyl
group preferably contains a cycloalkyl group that contains one or
two ring systems having from 3 to 10 (especially 3, 4, 5, 6 or 7)
ring carbon atoms, and one or two alkyl, alkenyl or alkynyl groups
having 1 or 2 to 6 carbon atoms. The expression aralkyl refers to a
group containing both aryl and also alkyl, alkenyl, alkynyl and/or
cycloalkyl groups in accordance with the above definitions, such
as, for example, an arylalkyl, arylalkenyl, arylalkynyl,
arylcycloalkyl, arylcycloalkenyl, alkylarylcycloalkyl and
alkylarylcycloalkenyl group. Specific examples of aralkyls are
toluene, xylene, mesitylene, styrene, benzyl chloride,
o-fluorotoluene, 1H-indene, tetraline, dihydronaphthalene,
indanone, phenylcyclopentyl, cumene, cyclohexylphenyl, fluorene and
indane. An aralkyl group preferably contains one or two aromatic
ring systems (1 or 2 rings) containing from 6 to 10 carbon atoms
and one or two alkyl, alkenyl and/or alkynyl groups containing from
1 or 2 to 6 carbon atoms and/or a cycloalkyl group containing 5 or
6 ring carbon atoms.
[0139] The expression heteroalkylcycloalkyl refers to
alkylcycloalkyl groups as defined above in which one or more,
preferably 1, 2 or 3, carbon atoms have been replaced independently
of each other by an oxygen, nitrogen, silicon, selenium, phosphorus
or sulfur atom (preferably by an oxygen, sulfur or nitrogen atom).
A heteroalkylcycloalkyl group preferably contains 1 or 2 ring
systems having from 3 to 10 (especially 3, 4, 5, 6 or 7) ring
atoms, and one or two alkyl, alkenyl, alkynyl or heteroalkyl groups
having from 1 or 2 to 6 carbon atoms. Examples of such groups are
alkylheterocycloalkyl, alkylheterocycloalkenyl,
alkenylheterocycloalkyl, alkynylheterocycloalkyl,
heteroalkylcycloalkyl, heteroalkylheterocycloalkyl and
heteroalkylheterocycloalkenyl, the cyclic groups being saturated or
mono-, di- or tri-unsaturated.
[0140] The expression heterocyclic ring refers to heteroaryl group
as defined above as well as to a cycloalkyl group or carbocyclic
ring as defined above in which one or more (preferably 1, 2 or 3)
ring carbon atoms, each independently, have been replaced by an
oxygen, nitrogen, silicon, selenium, phosphorus or sulfur atom,
preferably by an oxygen, sulfur or nitrogen atom. A heterocyclic
ring has preferably 1 or 2 ring(s) containing from 3 to 10,
especially 3, 4, 5, 6 or 7 ring atoms, preferably selected from C,
O, N and S. Examples are a aziridinyl, oxiranyl, thiiranyl,
oxaziridinyl, dioxiranyl, azetidinyl, oxetanyl, thietanyl,
diazetidinyl, dioxetanyl, dithietanyl, pyrrolidinyl,
tetrahydrofuranyl, thiolanyl, phospholanyl, silolanyl, azolyl,
thiazolyl, isothiazolyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl,
dioxolanyl, dithiolanyl, piperazinyl, morpholinyl,
thiopmorpholinyl, trioxanyl, azepanyl, oxepanyl, thiepanyl,
homopiperazinyl, or urotropinyl group.
[0141] The expression heteroaralkyl refers to an aralkyl group as
defined above in which one or more (preferably 1, 2, 3 or 4) carbon
atoms, each independently, have been replaced by an oxygen,
nitrogen, silicon, selenium, phosphorus, boron or sulfur atom
(preferably oxygen, sulfur or nitrogen), that is to say to a group
containing both aryl or heteroaryl, respectively, and also alkyl,
alkenyl, alkynyl and/or heteroalkyl and/or cycloalkyl and/or
heterocycloalkyl groups in accordance with the above definitions. A
heteroaralkyl group preferably contains one or two aromatic ring
systems (1 or 2 rings) containing from 5 or 6 to 10 ring carbon
atoms and one or two alkyl, alkenyl and/or alkynyl groups
containing 1 or 2 to 6 carbon atoms and/or a cycloalkyl group
containing 5 or 6 ring carbon atoms, wherein 1, 2, 3 or 4 of these
carbon atoms have been replaced by oxygen, sulfur or nitrogen
atoms.
[0142] Examples are arylheteroalkyl, arylheterocycloalkyl,
arylheterocycloalkenyl, arylalkylheterocycloalkyl,
arylalkenylheterocycloalkyl, arylalkynylheterocycloalkyl,
arylalkylheterocycloalkenyl, heteroarylalkyl, heteroarylalkenyl,
heteroarylalkynyl, heteroarylheteroalkyl, heteroarylcycloalkyl,
heteroarylcycloalkenyl, heteroarylheterocycloalkyl,
heteroarylheterocycloalkenyl, heteroarylalkylcycloalkyl,
heteroarylalkylheterocycloalkenyl, heteroarylheteroalkylcycloalkyl,
heteroarylheteroalkylcycloalkenyl and
heteroarylheteroalkylheterocycloalkyl groups, the cyclic groups
being saturated or mono-, di- or tri-unsaturated. Specific examples
are a tetrahydroisoquinolinyl, benzoyl, 2- or 3-ethylindolyl,
4-methylpyridino, 2-, 3- or 4-methoxyphenyl, 4-ethoxyphenyl, 2-, 3-
or 4-carboxyphenylalkyl group.
[0143] As already stated above, the expressions cycloalkyl,
heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aryl,
heteroaryl, aralkyl and heteroaralkyl also refer to groups in which
one or more hydrogen atoms of such groups have been replaced
independently of each other by fluorine, chlorine, bromine or
iodine atoms or by OH, .dbd.O, SH, .dbd.S, NH.sub.2, .dbd.NH,
N.sub.3 or NO.sub.2 groups.
[0144] The general term ring as used herein, unless defined
otherwise, includes cycloalkyl groups or carbocyclic rings,
heterocyclic rings, aryl groups, and heteroaryl groups.
[0145] The expressions "halo", "halogen"" or "halogen atom" as used
herein means fluorine, chlorine, bromine, or iodine, preferably
fluorine and/or chlorine.
[0146] The expression mono- or disaccharide, and derivatives
thereof as used herein means a carbohydrate or sugar belonging to
or derived from the group of monosaccharides or disaccharides.
[0147] Examples of mono-, disaccharides, and derivatives include
glucose, 3-O-methyl-glucose, 1-deoxy-glucose, 6-deoxy-glucose,
galactose, mannose, fructose, xylose, ribose, cellobiose, maltose,
lactose, gentiobiose, saccharose, trehalose and mannitol, sorbitol
and ribitol. Preferably, the saccharides are D-form saccharides,
e.g. D-glucose, 3-O-methyl-D-glucose, 1-deoxy-D-glucose, or
6-deoxy-D-glucose, D-galactose, D-mannose.
[0148] As used herein a wording defining the limits of a range of
length such as, e. g., "from 1 to 5" means any integer from 1 to 5,
i.e. 1, 2, 3, 4 and 5. In other words, any range defined by two
integers explicitly mentioned is meant to comprise and disclose any
integer defining said limits and any integer comprised in said
range.
[0149] The expression "--C(.dbd.O)O-motif" is used herein in order
to clearly define a group comprising an sp.sup.2-hybridized
carbonyl carbon attached (i) to any carbon or hetero atom and (ii)
to an oxygen which in turn can be attached to hydrogen or any other
chemical atom. The term "carboxyl group" is avoided for the
description of a "--C(.dbd.O)O-motif" because it could be mistaken
as describing the carboxylic acid only.
[0150] The term "in alpha position" is used to describe a directly
adjacent position, while the term "in beta position" indicates a
neighboring position of an atom or group A and an atom or group B,
characterized in that one further atom or group is localized
between A and B.
[0151] As used herein, the term oxamide refers to the arbitrarily
substituted organic compound comprising 2 carbonyl carbons and two
nitrogens, which compound is an arbitrarily substituted diamide
derived from any oxalic acid.
[0152] Those skilled in the art will readily recognize that some of
the n-3 PUFA analogues of general formula (I) of the present
invention represent "bioisosteres" of the naturally occurring
epoxymetabolites produced by cytochrome P450 (CYP) enzymes from
omega-3 (n-3) polyunsaturated fatty acids (PUFAs). A bioisostere is
a compound resulting from the exchange of an atom or of a group of
atoms with an alternative, broadly similar, atom or group of atoms,
thereby creating a new compound with similar biological properties
to the parent compound. Bioisosterism has, for example, been used
by medicinal chemists for improving desired biological or physical
properties of a compound, e.g. to attenuate toxicity, modify
activity, alter pharmacokinetics and/or metabolism of a compound.
For example, the replacement of a hydrogen atom with fluorine at a
site of metabolic oxidation in a compound may prevent such
metabolism from taking place. Because fluorine is similar in size
to the hydrogen atom the overall topology of the molecule is not
significantly affected, leaving the desired biological activity
unaffected. However, with a blocked pathway for metabolism, said
compound may have a longer half-life. Another example is the
bioisosteric replacement of carboxylic acid groups which has
resulted in analogues showing improved bioavailability, enhanced
blood-brain barrier penetration, increased activity, better
chemical stability and/or selectivity towards the target (see, e.g.
the textbook "The practice of medicinal chemistry", edited by
Camille Georges Wermuth, 3.sup.rd edition, Academic Press, 2008,
e.g. p. 303-310; Ballatore C. et al. "Carboxylic Acid
(Bio)Isosteres in Drug Design", Chem Med Chem 8, 385-395 (2013)).
Further, bioisosterism can also be used to provide a "prodrug" of a
compound, i.e. a compound that is initially administered to a
subject or patient in an inactive (or less active) form, and then
becomes modified in vivo to its active form through the normal
metabolic processes of the body. For example, conjugation of a
compound with lipid and/or sugar units has resulted in analogues
(prodrugs) showing increased drug delivery compared to the parent
compound (see, e.g. Wong A. and Toth I. "Lipid, Sugar and
Liposaccharide Based Delivery Systems", Current Medicinal Chemistry
8, 1123-1136 (2001)).
[0153] The n-3 PUFA analogues of general formula (I) of the present
invention can be prepared in a number of ways well known to one
skilled in the art of organic synthesis. For example, the compounds
of the present invention can be synthesized according to the
general reaction schemes shown below using synthetic methods known
in the art of synthetic organic chemistry, or variations thereon as
appreciated by those skilled in the art. Unless indicated
otherwise, all variables, e.g. n, k, R.sup.2 (also referred to as
R.sub.2), R.sup.6, R.sup.7, R.sup.8, R.sup.41, R.sup.42, R.sup.44
and R.sup.45, have the above defined meaning. As starting materials
reagents of standard commercial grade can be used without further
purification, or can be readily prepared from such materials by
routine methods. Those skilled in the art of organic synthesis will
recognize that starting materials and reaction conditions may be
varied including additional steps employed to produce compounds for
use encompassed by the present invention.
[0154] The compounds of the present invention are effective for
treating, reducing the risk of developing or preventing a disorder
associated with neovascularization and/or inflammation. In one
embodiment the disorder is a disorder associated with
neovascularization. In another embodiment the disorder is a
disorder associated with inflammation.
[0155] Examples of a disorder associated with inflammation include
inflammatory disorders, inflammation caused by other diseases
whatever type, etiology or pathogenesis, inflammation caused by
inflammatory diseases exemplified below and immunological
disorders.
[0156] In one embodiment the disorder associated with inflammation
is an inflammatory disorder. Examples of inflammatory disorders are
acute-phase reaction, local and systemic inflammation.
[0157] In one embodiment the disorder associated with inflammation
is an immunological disorder. Examples of immunological disorders
are hyperesthesia, autoimmune disorders, graft rejection in
transplantation, transplant toxicity, granulomatous
inflammation/tissue remodelling, myasthenia gravis,
immunosuppression, immune-complex diseases, over- and
underproduction of antibodies, and vasculitis.
[0158] In one embodiment the disorder associated with inflammation
is inflammation caused by other diseases whatever type, etiology or
pathogenesis, or inflammation caused by inflammatory diseases.
Examples of such conditions and diseases include inflammatory bowel
disease including Crohn's disease and ulcerative colitis, irritable
bowel syndrome, enterocolitis, liver diseases, pancreatitis,
nephritis, cystitis (interstitial cystitis), otitis media,
peridontitis, inflammatory skin disorders such as psoriasis,
eczema, atopic diseases, dermatitis, juvenile or adult onset
rheumatoid arthritis and gouty arthritis, ankylosing spondylitis,
adult onset or pediatric (systemic onset juvenile idiopathic
arthritis) Still's disease, psoriatic arthritis, osteoarthritis and
edema associated with burns, sprains or fracture, cerebral edema,
angioedema, vasculitis, diabetic vasculopathy, type I diabetes,
diabetic nephropathy, diabetic neuropathy, diabetic retinopathy,
post capillary resistance or diabetic syndromes associated with
insulitis (e.g. hyperglycemia, diuresis, proteinuria and increased
nitrite and kallikrein urinary excretion), gall bladder diseases,
multiple sclerosis, epilepsy, amyotrophic lateral sclerosis,
systemic inflammatory response syndrome (SIRS),
ischemia-reperfusion injury and atherosclerosis, septic shock,
inflammation caused by antihypovolemic and/or anti-hypotensive
agents, migraine, gingivitis, osteoporosis, benign prostatic
hyperplasia, hyperactive bladder, fibrotic diseases such as
pulmonary fibrosis, renal fibrosis, progressive sclerosis and
recurrent stricture formation in Crohn's disease, disorders of the
respiratory pathways in asthma, atopic or non-atopic asthma,
occupational asthma, exercise-induced bronchoconstriction,
bronchitis, pneumoconiosis including aluminosis, anhracosis,
asbestosis, chalicosis, ptilosis, siderosis, silicosis, tabaccosis
and byssinosis, chronic obstructive pulmonary disease including
emphysema, adult respiratory distress syndrome, pneumonia, allergic
rhinitis, vasomotor rhinitis and pleurisy, auto-inflammatory
diseases such as familial Mediterranean fever (FMF), tumor-necrosis
factor receptor associated periodic syndrome (TRAPS), neonatal
onset multisystem inflammatory disease (NOMID), familial cold
autoinflammatory syndrome (FCAS) including familial cold urticaria
(FCU), pyogenic arthritis pyoderma gangrenosum acne (PAPA) syndrome
and Muckle-Wells disease.
[0159] In a preferred embodiment the disorder associated with
neovascularization and/or inflammation is an ophthalmological
disorder associated with neovascularization, e.g., associated with
corneal, retinal, choroidal, uveal, or iris neovascularization
and/or an ophthalmological disorder associated with inflammation. A
preferred ophthalmological disorder associated with
neovascularization is age-related macular degeneration, e.g.,
neovascular (wet) AMD, or atrophic (dry) AMD. In this embodiment
the treatment results in blood vessel regression. In some
embodiments, the ophthalmological disorder associated with
neovascularization is retinopathy, preferably retinopathy of
prematurity (ROP); diabetic retinopathy; diabetic proliferative
retinopathy, retinal vein occlusion; e.g. branch retinal vein
occlusion, central retinal vein occlusion; sickle cell retinopathy;
and radiation retinopathy; Best's disease; or Stargardt's disease.
A preferred ophthalmological disorder associated with inflammation
is age-related macular degeneration.
[0160] In one embodiment the disorder associated with
neovascularization and/or inflammation is not a cardiovascular
disease.
[0161] In a preferred embodiment, the compound or composition for
use according to the invention is administered orally, topically,
subcutaneously, intravitrealy, intramuscularly, intraperitoneally,
intravenously, or intranasally, preferably orally or
intraveneously, more preferably orally or intraperitoneally. The
preferred delivery route of ocular medications to the eye for the
treatment of an ophthalmological disorder is topical, local ocular
(e.g., subconjunctival, intravitreal, retrobulbar, intracameral),
and systemic. The latter is preferably achieved through oral,
intramuscular or intravenous administration.
[0162] It is further preferred that the compound or composition for
use according to the invention 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.
[0163] The compound or composition for use according to the
invention may further comprise at least one compound of formula (I)
and, optionally, one or more carrier substances, e.g. cyclodextrins
such as hydroxypropyl .beta.-cyclodextrin, micelles or liposomes,
excipients and/or adjuvants. It may additionally comprise, for
example, one or more of water, buffers such as, e.g., neutral
buffered saline or phosphate buffered saline, ethanol, mineral oil,
vegetable oil, dimethylsulfoxide, carbohydrates such as e.g.,
glucose, mannose, sucrose or dextrans, mannitol, proteins,
adjuvants, polypeptides or amino acids such as glycine,
antioxidants, chelating agents such as EDTA or glutathione and/or
preservatives. Furthermore, one or more other active ingredients
may, but need not, be included compositions for us provided herein.
For instance, the compounds of the invention may advantageously be
employed in combination with an antibiotic, anti-fungal, or
anti-viral agent, an anti-histamine, a non-steroidal
anti-inflammatory drug, a disease modifying anti-rheumatic drug, an
anti-inflammatory drug to treat an autoimmune disease, a cytostatic
drug, an antiangiogenic drug or mixtures of the aforementioned.
Preferably the drug is an antiangiogenic drug, more preferably an
inhibitor of Vascular Endothelial Growth Factor (VEGF) or Vascular
Endothelial Growth Factor Receptor (VEGFR).
[0164] The compositions for use may be formulated for any
appropriate route of administration, including, for example,
topical such as, e.g., transdermal or ocular, oral, buccal, nasal,
vaginal, rectal or parenteral administration. The term parenteral
as used herein includes subcutaneous, intradermal, intravascular
such as, e.g., intravenous, intramuscular, spinal, intracranial,
intrathecal, intraocular, periocular, intraorbital, intrasynovial,
intraperitoneal and local ocular (e.g., subconjunctival,
intravitreal, retrobulbar, intracameral) injection, as well as any
similar injection or infusion technique. In certain embodiments,
compositions in a form suitable for oral use are preferred. Such
forms include, for example, tablets, troches, lozenges, aqueous or
oily suspensions, dispersible powders or granules, emulsion, hard
or soft capsules, or syrups or elixirs. Within yet other
embodiments, compositions provided herein may be formulated as a
lyophilizate.
[0165] Compositions intended for oral use may further comprise one
or more components such as sweetening agents, flavoring agents,
coloring agents and/or preserving agents in order to provide
appealing and palatable preparations. Tablets contain the active
ingredient in admixture with physiologically acceptable excipients
that are suitable for the manufacture of tablets. Such excipients
include, for example, inert diluents such as, e.g., calcium
carbonate, sodium carbonate, lactose, calcium phosphate or sodium
phosphate, granulating and disintegrating agents such as, e.g.,
corn starch or alginic acid, binding agents such as, e.g., starch,
gelatin or acacia, and lubricating agents such as, e.g., magnesium
stearate, stearic acid or talc. The tablets may be uncoated or they
may be coated by known techniques to delay disintegration and
absorption in the gastrointestinal tract and thereby provide a
sustained action over a longer period. For example, a time delay
material such as glyceryl monosterate or glyceryl distearate may be
employed. Methods for preparing such compositions are known (see,
for example, H. C. Ansel and N. G. Popovish, Pharmaceutical Dosage
Forms and Drug Delivery Systems, 5th ed., Lea and Febiger
(1990)).
[0166] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent such as, e.g., calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium such as, e.g.,
peanut oil, liquid paraffin or olive oil.
[0167] Aqueous suspensions contain the active ingredient(s) in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients include suspending agents such as,
e.g., sodium carboxymethylcellulose, methylcellulose,
hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone,
gum tragacanth and gum acacia; and dispersing or wetting agents
such as, e.g., naturally-occurring phosphatides such as lecithin,
condensation products of an alkylene oxide with fatty acids such as
polyoxyethylene stearate, condensation products of ethylene oxide
with long chain aliphatic alcohols such as
heptadecaethyleneoxycetanol, condensation products of ethylene
oxide with partial esters derived from fatty acids and a hexitol
such as polyoxyethylene sorbitol monooleate, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and hexitol anhydrides such as polyethylene sorbitan
monooleate. Aqueous suspensions may also comprise one or more
preservatives, for example ethyl, or n-propyl p-hydroxybenzoate,
one or more coloring agents, one or more flavoring agents, and one
or more sweetening agents, such as sucrose or saccharin.
[0168] Oily suspensions may be formulated by suspending the active
ingredients in a vegetable oil such as, e.g., arachis oil, olive
oil, sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent such
as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such
as those set forth above, and/or flavoring agents may be added to
provide palatable oral preparations. Such suspensions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0169] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, such as sweetening,
flavoring and coloring agents, may also be present.
[0170] Compositions for use may also be in the form of oil-in-water
emulsions. The oily phase may be a vegetable oil such as, e.g.,
olive oil or arachis oil, a mineral oil such as, e.g., liquid
paraffin, or a mixture thereof. Suitable emulsifying agents include
naturally-occurring gums such as, e.g., gum acacia or gum
tragacanth, naturally-occurring phosphatides such as, e.g., soy
bean lecithin, and esters or partial esters derived from fatty
acids and hexitol, anhydrides such as, e.g., sorbitan monoleate,
and condensation products of partial esters derived from fatty
acids and hexitol with ethylene oxide such as, e.g.,
polyoxyethylene sorbitan monoleate. An emulsion may also comprise
one or more sweetening and/or flavoring agents.
[0171] Syrups and elixirs may be formulated with sweetening agents,
such as glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also comprise one or more demulcents,
preservatives, flavoring agents and/or coloring agents.
[0172] Compounds for use according to the invention may be
formulated for local or topical administration, such as for topical
application to the skin or mucous membranes, such as in the eye.
Formulations for topical administration typically comprise a
topical vehicle combined with active agent(s), with or without
additional optional components. Suitable topical vehicles and
additional components are well known in the art, and it will be
apparent that the choice of a vehicle will depend on the particular
physical form and mode of delivery. Topical vehicles include water;
organic solvents such as alcohols such as, e.g., ethanol or
isopropyl alcohol or glycerin; glycols such as, e.g., butylene,
isoprene or propylene glycol; aliphatic alcohols such as, e.g.,
lanolin; mixtures of water and organic solvents and mixtures of
organic solvents such as alcohol and glycerin; lipid-based
materials such as fatty acids, acylglycerols including oils, such
as, e.g., mineral oil, and fats of natural or synthetic origin,
phosphoglycerides, sphingolipids and waxes; protein-based materials
such as collagen and gelatin; silicone-based materials, both
non-volatile and volatile; and hydrocarbon-based materials such as
microsponges and polymer matrices. A composition may further
include one or more components adapted to improve the stability or
effectiveness of the applied formulation, such as stabilizing
agents, suspending agents, emulsifying agents, viscosity adjusters,
gelling agents, preservatives, antioxidants, skin penetration
enhancers, moisturizers and sustained release materials. Examples
of such components are described in Martindale-The Extra
Pharmacopoeia (Pharmaceutical Press, London 1993) and Martin (ed.),
Remington's Pharmaceutical Sciences. Formulations may comprise
microcapsules, such as hydroxymethylcellulose or
gelatin-microcapsules, liposomes, albumin microspheres,
microemulsions, nanoparticles or nanocapsules.
[0173] A topical formulation for use may be prepared in a variety
of physical forms including, for example, solids, pastes, creams,
foams, lotions, gels, powders, aqueous liquids, emulsions, sprays,
eye-drops and skin patches. The physical appearance and viscosity
of such forms can be governed by the presence and amount of
emulsifier(s) and viscosity adjuster(s) present in the formulation.
Solids are generally firm and non-pourable and commonly are
formulated as bars or sticks, or in particulate form; solids can be
opaque or transparent, and optionally can contain solvents,
emulsifiers, moisturizers, emollients, fragrances, dyes/colorants,
preservatives and other active ingredients that increase or enhance
the efficacy of the final product. Creams and lotions are often
similar to one another, differing mainly in their viscosity; both
lotions and creams may be opaque, translucent or clear and often
contain emulsifiers, solvents, and viscosity adjusting agents, as
well as moisturizers, emollients, fragrances, dyes/colorants,
preservatives and other active ingredients that increase or enhance
the efficacy of the final product. Gels can be prepared with a
range of viscosities, from thick or high viscosity to thin or low
viscosity. These formulations, like those of lotions and creams,
may also contain solvents, emulsifiers, moisturizers, emollients,
fragrances, dyes/colorants, preservatives and other active
ingredients that increase or enhance the efficacy of the final
product. Liquids are thinner than creams, lotions, or gels and
often do not contain emulsifiers. Liquid topical products often
contain solvents, emulsifiers, moisturizers, emollients,
fragrances, dyes/colorants, preservatives and other active
ingredients that increase or enhance the efficacy of the final
product.
[0174] Suitable emulsifiers for use in topical formulations
include, but are not limited to, ionic emulsifiers, cetearyl
alcohol, non-ionic emulsifiers like polyoxyethylene oleyl ether,
PEG-40 stearate, ceteareth-12, ceteareth-20, ceteareth-30,
ceteareth alcohol, PEG-100 stearate and glyceryl stearate. Suitable
viscosity adjusting agents include, but are not limited to,
protective colloids or non-ionic gums such as
hydroxyethylcellulose, xanthan gum, magnesium aluminum silicate,
silica, microcrystalline wax, beeswax, paraffin, and cetyl
palmitate. A gel composition may be formed by the addition of a
gelling agent such as chitosan, methyl cellulose, ethyl cellulose,
polyvinyl alcohol, polyquaterniums, hydroxyethylcellulose,
hydroxypropylcellulose, hydroxypropylmethylcellulose, carbomer or
ammoniated glycyrrhizinate. Suitable surfactants include, but are
not limited to, nonionic, amphoteric, ionic and anionic
surfactants. For example, one or more of dimethicone copolyol,
polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80,
lauramide DEA, cocamide DEA, and cocamide MEA, oleyl betaine,
cocamidopropyl phosphatidyl PG-dimonium chloride, and ammonium
laureth sulfate may be used within topical formulations.
[0175] Suitable preservatives include, but are not limited to,
antimicrobials such as methylparaben, propylparaben, sorbic acid,
benzoic acid, and formaldehyde, as well as physical stabilizers and
antioxidants such as vitamin E, sodium ascorbate/ascorbic acid and
propyl gallate. Suitable moisturizers include, but are not limited
to, lactic acid and other hydroxy acids and their salts, glycerin,
propylene glycol, and butylene glycol. Suitable emollients include
lanolin alcohol, lanolin, lanolin derivatives, cholesterol,
petrolatum, isostearyl neopentanoate and mineral oils. Suitable
fragrances and colors include, but are not limited to, FD&C Red
No. 40 and FD&C Yellow No. 5. Other suitable additional
ingredients that may be included in a topical formulation include,
but are not limited to, abrasives, absorbents, anti-caking agents,
anti-foaming agents, anti-static agents, astringents such as, e.g.,
witch hazel, alcohol and herbal extracts such as chamomile extract,
binders/excipients, buffering agents, chelating agents, film
forming agents, conditioning agents, propellants, opacifying
agents, pH adjusters and protectants.
[0176] An example of a suitable topical vehicle for formulation of
a gel is: hydroxypropylcellulose (2.1%); 70/30 isopropyl
alcohol/water (90.9%); propylene glycol (5.1%); and Polysorbate 80
(1.9%). An example of a suitable topical vehicle for formulation as
a foam is: cetyl alcohol (1.1%); stearyl alcohol (0.5%); Quaternium
52 (1.0%); propylene glycol (2.0%); Ethanol 95 PGF3 (61.05%);
deionized water (30.05%); P75 hydrocarbon propellant (4.30%). All
percents are by weight.
[0177] Typical modes of delivery for topical compositions include
application using the fingers; application using a physical
applicator such as a cloth, tissue, swab, stick or brush; spraying
including mist, aerosol or foam spraying; dropper application;
sprinkling; soaking; and rinsing. Controlled release vehicles can
also be used, and compositions may be formulated for transdermal
administration as a transdermal patch.
[0178] A composition for use may be formulated as inhaled
formulations, including sprays, mists, or aerosols. Such
formulations are particularly useful for the treatment of asthma or
other respiratory conditions. For inhalation formulations, the
compounds provided herein may be delivered via any inhalation
methods known to those skilled in the art. Such inhalation methods
and devices include, but are not limited to, metered dose inhalers
with propellants such as CFC or HFA or propellants that are
physiologically and environmentally acceptable. Other suitable
devices are breath operated inhalers, multidose dry powder inhalers
and aerosol nebulizers. Aerosol formulations for use in the subject
method typically include propellants, surfactants and co-solvents
and may be filled into conventional aerosol containers that are
closed by a suitable metering valve.
[0179] Inhalant compositions may comprise liquid or powdered
compositions containing the active ingredient that are suitable for
nebulization and intrabronchial use, or aerosol compositions
administered via an aerosol unit dispensing metered doses. Suitable
liquid compositions comprise the active ingredient in an aqueous,
pharmaceutically acceptable inhalant solvent, e.g., isotonic saline
or bacteriostatic water. The solutions are administered by means of
a pump or squeeze-actuated nebulized spray dispenser, or by any
other conventional means for causing or enabling the requisite
dosage amount of the liquid composition to be inhaled into the
patient's lungs. Suitable formulations, wherein the carrier is a
liquid, for administration, as for example, a nasal spray or as
nasal drops, include aqueous or oily solutions of the active
ingredient.
[0180] Formulations or compositions suitable for nasal
administration, wherein the carrier is a solid, include a coarse
powder having a particle size, for example, in the range of 20 to
500 microns which is administered in the manner in which snuff is
administered, i.e., by rapid inhalation through the nasal passage
from a container of the powder held close up to the nose. Suitable
powder compositions include, by way of illustration, powdered
preparations of the active ingredient thoroughly intermixed with
lactose or other inert powders acceptable for intrabronchial
administration. The powder compositions can be administered via an
aerosol dispenser or encased in a breakable capsule which may be
inserted by the patient into a device that punctures the capsule
and blows the powder out in a steady stream suitable for
inhalation.
[0181] Compositions for use may also be prepared in the form of
suppositories such as e.g., for rectal administration. Such
compositions can be prepared by mixing the drug with a suitable
non-irritating excipient that is solid at ordinary temperatures but
liquid at the rectal temperature and will therefore melt in the
rectum to release the drug. Suitable excipients include, for
example, cocoa butter and polyethylene glycols.
[0182] Compositions for use may be formulated as sustained release
formulations such as, i.e., a formulation such as a capsule that
creates a slow release of modulator following administration. Such
formulations may generally be prepared using well known technology
and administered by, for example, oral, rectal or subcutaneous
implantation, or by implantation at the desired target site.
Carriers for use within such formulations are biocompatible, and
may also be biodegradable; preferably the formulation provides a
relatively constant level of modulator release. The amount of
modulator contained within a sustained release formulation depends
upon, for example, the site of implantation, the rate and expected
duration of release and the nature of the condition to be treated
or prevented.
[0183] It will be understood, however, that the specific dose level
for any particular patient will depend upon a variety of factors
including the activity of the specific compound employed, the age,
body weight, general health, sex, diet, time of administration,
route of administration, and rate of excretion, drug combination,
i.e. other drugs being used to treat the patient, and the severity
of the particular disease undergoing therapy.
[0184] Preferred compounds of the invention will have certain
pharmacological properties. Such properties include, but are not
limited to oral bioavailability, such that the preferred oral
dosage forms discussed above can provide therapeutically effective
levels of the compound in vivo.
[0185] n-3 PUFA derivatives provided herein are preferably
administered to a patient such as, e.g., a human, orally or
parenterally, and are present within at least one body fluid or
tissue of the patient.
[0186] As used herein, the term "treatment" encompasses both any
type of disease-modifying treatment and including symptomatic
treatment, i.e., a treatment after the onset of symptoms, either of
which may be prophylactic. However, disease-modifying treatment may
involve administration before the onset of symptoms, in order to
prevent, at least delay or reduce the severity of symptoms after
onset. A disease-modifying treatment may also be therapeutic, i.e.,
after the onset of symptoms, in order to reduce the severity and/or
duration of symptoms. A treatment after onset of symptoms may also
simply involve stopping progressing of the disease (stable
disease). In certain embodiment, the n-3 PUFA derivatives provided
herein are administered prophylactically, i.e., before the onset of
the disease and/or symptoms, ideally, but not necessarily, to
actually prevent the diseases and/or symptoms. It is to be
understood that the term prophylaxis and prophylactic in the
context of the present invention, simply describes that the
compound(s) of the present invention are administered before the
onset of symptoms. A prophylactic administration may an
administration before the onset of symptoms that are clearly
associated with a disease discussed herein: the n-3 PUFA
derivatives provided herein may, e.g., be administered to a subject
prophylactically when he or she displays certain conditions that
may indicate a propensity to develop one of the conditions or
diseases that can be treated with one of the n-3 PUFA derivatives
of the present invention. Such indicative conditions are, e.g. high
blood pressure or diabetes. Such a prophylactic treatment is called
primary prophylaxis. In another embodiment, the n-3 PUFA
derivatives provided herein may be administered to a subject
prophylactically when he or she has previously suffered from a
condition or disease that can be treated with the n-3 PUFA
derivatives of the present invention, but currently does not
display any symptoms. Such a prophylactic treatment is called
secondary prophylaxis. Patients receiving the n-3 PUFA derivatives
for the purpose of primary or secondary prophylaxis are considered
to be in need of such a treatment. Patients may include but are not
limited to mammals, especially humans, domesticated companion
animals such as dogs, cats, horses, and livestock such as cattle,
pigs, sheep, with dosages as described herein.
[0187] The activity of the n-3 PUFA analogues according to the
invention can, for example, be determined in appropriate in vitro
and/or in vivo assays. For instance, the biological activity of the
n-3 PUFA analogues according to the present invention may be
determined using the established cell model of Kang and Leaf (Proc
Natl Acad Sci USA, 1994. 91(21): p. 9886-90.) known to those
skilled in the art.
[0188] The following figure and examples serve to illustrate the
invention and are not intended to limit the scope of the invention
as described in the appended claims.
FIGURES
[0189] FIG. 1: Efficacy of compound with the structure according to
Formula (VI), which is referred to as "Compound 1" in a laser
induced choroidal neovascularization model in rat evaluated by
measuring vascular leakage using fluorescence angiography:
Fluorescence angiography grading data (mean.+-.SD) on day 14 and 21
after induction of choroidal neovascularization (laser burn),
grading scale: 0-3 arbitrary unit. The leakage of fluorescein was
evaluated in the fluorescence angiograms by two independent
examiners masked to the study groups and graded as follows: Score
0: no leakage; Score 1: slightly stained; Score 2: moderately
stained; Score 3: strongly stained, n=10 animals/group with 6
lesions per animal, statistic: Kruskal-Wallis-test and Dunn's
multiple comparisons test, *p<0.05, po: oral administration, ip:
intraperitoneal administration, IVT: intravitreal administration,
FA: Fluorescence angiography
[0190] FIG. 2: FIG. 2 shows the anti-inflammatory effect of a
metabolically robust analog of 17,18-EEQ (Comp-02) on HL-1
cardiomyocytes. A cardiomyocyte cell line was used (mouse derived
immortalized cardiomyocytes, HL-1 cells). Cells were either treated
with vehicle (0.01% ethanol) or different concentrations of test
compound (Comp-02: cE=10 nM, 100 nM or 1 .mu.M). Simultaneously,
the cells were challenged with 1 .mu.g/mL lipopolysaccharide (LPS).
After 24 h of incubation, the cells were processed to measure
viability.
[0191] FIG. 3: FIG. 3 shows the anti-inflammatory effect of a
metabolically robust analog of 17,18-EEQ (Comp-02) on HL-1
cardiomyocytes. A cardiomyocyte cell line was used (mouse derived
immortalized cardiomyocytes, HL-1 cells). Cells were either treated
with vehicle (0.01% ethanol) or different concentrations of test
compound (Comp-02: cE=10 nM, 100 nM or 1 .mu.M). Simultaneously,
the cells were challenged with 1 .mu.g/mL lipopolysaccharide (LPS).
After 24 h of incubation, the cells were processed to measure
release of the pro-inflammatory cytokine TNF alpha.
[0192] FIG. 4: FIG. 4 shows the inhibitory effect of a
metabolically robust analog of 17,18-EEQ (Comp-02) on cardiac
inflammation in a rat model of severe hypertension and end-organ
damage. Modulation of inflammation was measured with macrophages
infiltration via ED1 staining as a marker for inflammation. The
values shown are expressed as median with interquartile range.
Count values are pooled in bins of 20 view fields. The results show
that dTGR animals have significant higher amounts of infiltrated
macrophages in the cardiac tissue compared to non-treated SD
animals. However, Comp-02 treatment led to a significant reduction
of macrophage infiltration (ED1 positive cells) in dTGR animals
compared to vehicle treated dTGRs.
[0193] FIG. 5: FIG. 5 shows the inhibitory effect of a
metabolically robust analog of 17,18-EEQ (Comp-02) on renal
inflammation in a rat model of severe hypertension and end-organ
damage. Modulation of inflammation was measured with macrophages
infiltration via ED1 staining as a marker for inflammation. The
values shown are expressed as median with interquartile range.
Count values are pooled in bins of 20 view fields. The results show
that dTGR animals have significant higher amounts of infiltrated
macrophages in the renal tissue compared to non-treated SD animals.
However, Comp-02 treatment led to a significant reduction of
macrophage infiltration (ED1 positive cells) in dTGR animals
compared to vehicle treated dTGRs.
EXAMPLE 1
Synthesis of Compounds
[0194] In the following the synthesis of selected compounds of the
invention is illustrated.
Compound 1 (Comp-01)
[0195] 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
##STR00024## ##STR00025##
[0196] General Method
[0197] 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 50.times.2.0 mm, 5 um) or Agilent 1200 HPLC, 1956 MSD (Column:
Shim-pack XR-ODS 30.times.3.0 mm, 2.2 um) operating in ES (+)
ionization mode. Chromatographic purifications were by flash
chromatography using 100.about.200 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
##STR00026##
TABLE-US-00001 [0198] Other Reagent MW. Amount Mmol ratio Info.
Compound 1 136.53 100 g 732.44 1 MeNH.sub.2--HCl 67.52 64.29 g
952.17 1.3 Et.sub.3N 101.19 185.29 g 1830 2.5 THF 2 L Product
131.13 70 g 533.82 Yield: (compound 2) 73%
[0199] 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.degree. C. And the mixture was stirred
at 25.degree. 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.
[0200] 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) 4.36.about.4.24 (q, J=8 Hz, 2H), 2.93.about.2.85 (d, J=4 Hz,
3H), 1.38.about.1.30 (t, J=8 Hz, 3H)
General Procedure for Preparation of Compound 4
##STR00027##
TABLE-US-00002 [0201] Other Reagent MW. amount mmol ratio 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 (compound 5) 77.3%
[0202] 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.degree. 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.degree. 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.
[0203] 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)
7.86.about.7.79 (m, 2H), 7.72.about.7.67 (m, 2H), 3.73.about.3.66
(t, J=8 Hz, 2H), 2.27.about.2.20 (m, 2H), 1.95.about.1.91 (t, J=4
Hz, 1H), 1.85.about.1.75 (m, 2H), 1.61.about.1.52 (m, 2H)
General Procedure for Preparation of Compound 6
##STR00028##
TABLE-US-00003 [0204] Other Reagent MW. amount mmol ratio 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 353.15 118.6
g 335.8 Yield: (Compound 6) 86%
[0205] 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.degree. 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.
[0206] 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)
7.87.about.7.82 (m, 2H), 7.74.about.7.69 (m, 2H), 3.74.about.3.67
(t, J=8 Hz, 2H), 2.45.about.2.39 (t, J=8 Hz, 2H), 1.84.about.1.74
(m, 2H), 1.61.about.1.52 (m, 2H)
General Procedure for Preparation of Compound 7
##STR00029##
TABLE-US-00004 [0207] Other Reagent MW. amount mmol ratio Info.
Compound 6 353.15 157 g 444.57 1 BH.sub.3.cndot.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 355.17 135 g 380.1 Yield: (compound 5) 85%
[0208] 2-methylbut-2-ene (87.30 g, 1.24 mol, 2.80 eq.) was added
over 30 min to a 0.degree. 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.degree. C. and stirred for 90 min. After
re-cooling to 0.degree. 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.degree. C. After 2 h, the reaction was
cooled again to 0.degree. C. where upon glacial AcOH (260 mL) was
added slowly over 30 min (Caution: gas evolution) and stirred at
25.degree. 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.
[0209] 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)
7.88.about.7.80 (m, 2H), 7.75.about.7.67 (m, 2H), 6.24.about.6.11
(m, 2H), 3.74.about.3.66 (t, J=8 Hz, 2H), 2.24.about.2.15 (q, J=8
Hz, 2H), 1.78.about.1.67 (m, 2H), 1.55.about.1.44 (m, 2H)
General Procedure for Preparation of Compound 8
##STR00030##
TABLE-US-00005 [0210] Other Reagent MW. amount mmol ratio Info.
Compound 7 355.17 138 g 388.55 1 N.sub.2H.sub.4.cndot.H.sub.2O
50.06 97.25 g 1940 5 MeOH 2 L Product 225.07 81 g 683.79 Yield:
(compound 8) 92%
[0211] 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.degree. C. and stirred at
25.degree. 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.
[0212] 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)
6.19.about.6.07 (m, 2H), 2.73.about.2.59 (m, 2H), 2.20.about.2.05
(m, 2H), 1.75.about.1.55 (m, 2H), 1.51.about.1.36 (m, 4H)
General Procedure for Preparation of Compound 9
##STR00031##
TABLE-US-00006 [0213] Other Reagent MW. amount mmol ratio 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 310.13 90 g
232.16 Yield: (compound 9) 57%
[0214] 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.degree. 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.
[0215] 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)
7.57.about.7.37 (s, 2H), 6.25.about.6.20 (d, J=8 Hz, 1H),
6.18.about.6.11 (q, J=8 Hz, 1H), 3.37-3.30 (q, J=8 Hz, 2H),
2.93.about.2.88 (d, J=4 Hz, 3H), 2.21.about.2.13 (m, 2H),
1.66.about.1.56 (m, 2H), 1.53.about.1.43 (m, 2H)
General Procedure for Preparation of Compound 12
##STR00032##
TABLE-US-00007 [0216] Other Reagent MW. amount mmol ratio 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%
[0217] 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.degree.
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.
[0218] 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)
5.86.about.5.72 (m, 1H), 5.03.about.5.86 (m, 2H), 4.24.about.4.17
(q, J=8 Hz, 2H), 4.07.about.4.01 (s, 2H), 3.54.about.3.47 (t, J=8
Hz, 2H), 2.09.about.1.98 (m, 2H), 1.68.about.1.55 (m, 2H),
1.45.about.1.32 (m, 4H), 1.30.about.1.25 (t, J=8 Hz, 3H)
General Procedure for Preparation of Compound 13
##STR00033##
TABLE-US-00008 [0219] Other Reagent MW. amount mmol ratio 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%
[0220] 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.degree. C. After
stirring at 25.degree. 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.degree. C. for 5 h. LCMS show the reaction was completed. After
cooling to 25.degree. 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.
[0221] 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) 0.57.about.7.38 (s, 1H),
5.41.about.5.25 (m, 2H), 4.25.about.4.17 (q, J=8 Hz, 2H),
4.07.about.4.02 (s, 2H), 3.54.about.3.47 (t, J=8 Hz, 2H),
3.34.about.3.26 (q, J=8 Hz, 2H), 2.92.about.2.87 (d, J=8 Hz, 3H),
2.08.about.1.94 (m, 4H), 1.65.about.1.51 (m, 4H), 1.43.about.1.23
(m, 13H)
General Procedure for Preparation of Comp-02
##STR00034##
TABLE-US-00009 [0222] Other Reagent MW. amount mmol ratio 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%
[0223] 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.degree. C. and then the reaction
mixture was stirred at 0-25.degree. 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)
[0224] 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),
5.40.about.5.32 (m, 2H), 4.11 (s, 2H), 3.59.about.3.55 (t, J=6.4
Hz, 2H), 3.35.about.3.32 (t, J=6.8 Hz, 2H), 2.92.about.2.91 (d,
J=5.2 Hz, 3H), 2.07.about.2.00 (m, 4H), 1.64-1.59 (m, 4H),
1.42.about.1.32 (m, 10H); .sup.13C NMR (CDCl3, 100 MHz) .delta.
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
##STR00035## ##STR00036##
[0226] Synthesis of 2-(hex-5-yn-1-yl)isoindoline-1,3-dione (2):
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.degree. 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.RTM. 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
##STR00037##
[0227] Synthesis of 2-(6-iodohex-5-yn-1-yl)isoindoline-1,3-dione
(3): 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 (2.times.50 mL). The
ethereal extracts were washed with brine (3.times.60 mL) (Note:
biphasic mixture turned brown). The combined aqueous phases were
re-extracted with Et.sub.2O (2.times.50 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.RTM. 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.degree. C. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
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) .delta.
168.62, 134.14, 132.30, 123.44, 94.04, 37.60, 27.91, 25.89, 20.60,
-6.27.
##STR00038##
[0228] Synthesis of 2-(6-iodohex-5(Z)-en-1-yl)isoindoline-1,3-dione
(4): Following literature precedent,.sup.4 neat 2-methyl-2-butene
(4.2 mL, 2.8 equiv) was added over 5 min to a 0.degree. 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.degree. 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.degree. 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 (2.times.40 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.RTM. 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.
##STR00039##
[0229] Synthesis of 6-iodohex-5(Z)-en-1-amine (5): Following
literature precedent,' 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
mL.times.2). The combined ethereal extracts were washed with cold
1N HCl solution (20 mL.times.2). The combined aqueous washes were
adjusted to pH 8 with dilute, aq. NaOH. The solution was extracted
with Et.sub.2O (30 mL.times.2), 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) .delta.
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).
##STR00040##
[0230] Synthesis of
N.sup.1-(6-iodohex-5(Z)-en-1-yl)-N.sup.2-methyloxalamide (7):
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.degree. C. After 20 h, the brown
solution was cooled to rt and concentrated in vacuo. Purification
of the residue using a Teledyne Isco Combiflash.RTM. 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.degree. C. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 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) .delta. 160.47, 159.70, 140.43, 83.07, 39.40, 34.11,
28.61, 26.15, 25.11.
##STR00041##
[0231] Synthesis of ethyl 2-(oct-7-en-1-yloxy)acetate (10):
Following literature precedent,` 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.RTM. 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) .delta. 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) .delta. 170.70, 138.99, 114.48, 71.97, 68.48, 60.86,
33.84, 29.55, 28.84, 25.63, 14.34.
##STR00042##
[0232] Synthesis of ethyl
2-((13-(2-(methylamino)-2-oxoacetamido)tridec-8(Z)-en-1-yl)oxy)acetate
(11): 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.degree. 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.RTM. 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.
[0233] TLC: 50% EtOAc/hexanes, R.sub.f.about.0.49. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 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) .delta. 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.
##STR00043##
[0234] Synthesis of
2-((13-(2-(methylamino)-2-oxoacetamido)tridec-8(Z)-en-1-yl)oxy)acetic
acid (12): 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.degree. 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 mL.times.3). 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.RTM. 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.degree. C.
[0235] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 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) .delta. 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.
##STR00044##
[0236] Synthesis of Comp-03: 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 mL.times.2). 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.RTM. 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.degree. C.
[0237] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 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) .delta. 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)
[0238] 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
##STR00045## ##STR00046##
[0239] General Procedure for Preparation of Compound 4-2
##STR00047##
TABLE-US-00010 [0240] Other Reagent MW. amount Mol Ratio 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. Cul 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: 189.25 21.0 g
99.9 mmol Yield: (Cpd. 4-2) 73%
[0241] 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.degree. C. and stirred at 25.degree. 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.
[0242] .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
##STR00048##
TABLE-US-00011 [0243] Other Reagent MW. amount Mol Ratio Info. Cpd.
4-2 189.25 21.0 g 111 mmol 1.0 eq. Pd/C 500 mg MeOH 500 mL Product:
193.28 17.0 g 83.6 mmol Yield: (Cpd. 4-3) 75%
[0244] 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.degree. 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.
[0245] .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
##STR00049##
TABLE-US-00012 [0246] Other Reagent MW. amount Mol Ratio 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.
Con.cndot.H.sub.2SO.sub.4 98.08 30.2 g 308 mmol 3.5 eq. H.sub.2O
560 mL Product: 304.17 17.0 g 50.3 mmol Yield: (Cpd. 4-4) 57%
[0247] 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.degree. 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.degree. C. and stirred at 0.degree. 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.degree. C. and the resulting suspension was warmed to 25.degree.
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.
[0248] .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'
##STR00050##
TABLE-US-00013 [0249] Other Reagent MW. amount Mol Ratio 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: 418.31 5.40 g 12.3 mmol Yield: (Cpd.
4-5') 37%
[0250] 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.degree. C.,
then the mixture was stirred at 25.degree. 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.
[0251] .sup.1H NMR: ET5008-26-P1b1 400 MHz CDCl.sub.3;
.quadrature.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
##STR00051##
TABLE-US-00014 [0252] Other Reagent MW. amount Mol Ratio 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. Cul 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: 459.62 3.00 g 6.20 mmol Yield: (Cpd. 4-6)
48%
[0253] 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.degree. C. under N.sub.2 and stirred at
25.degree. 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.
[0254] .sup.1H NMR: ET5008-32-P1b1 400 MHz CDCl.sub.3;
.quadrature.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
##STR00052##
TABLE-US-00015 [0255] Other Reagent MW. amount Mol Ratio Info. Cpd.
4-6 459.62 3.00 g 6.53 mmol 1.0 eq. Pd/C 200 mg MeOH 20.0 mL
Product: 463.65 2.50 g 4.91 mmol Yield: (Cpd. 4-7) 75%
[0256] 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.degree. C.
under 50 psi of H2 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.
[0257] .sup.1H NMR: ET5008-33-P1b1 400 MHz CDCl.sub.3;
.quadrature.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
##STR00053##
TABLE-US-00016 [0258] Other Reagent MW. amount Mol Ratio Info. Cpd.
4-7 463.65 1.00 g 2.16 mmol 1.0 eq. HCl/EtOAc 30.0 mL 4N Product:
343.89 800 mg 2.33 mmol crude (Cpd. 4-10)
[0259] A mixture of Cpd.4-7 (1.00 g, 2.16 mmol, 1.0 eq) in
HCl/EtOAc (30.0 mL) at 50.degree. C. and stirred at 50.degree. 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
##STR00054##
TABLE-US-00017 [0260] Other Reagent MW. amount Mol Ratio 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: 392.49 370 mg 933 umol Yield: Comp-05 40%
[0261] 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.degree. C. Then the
solution was stirred at 60.degree. 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:
TABLE-US-00018 [0262] Column Luna C18 100*30 5 u Condition 0.05%
HCl-can Begin B 30 End B 60 Gradient Time 12 min 100% B Hold Time 4
min Flow Rate 25 mL/min Injection 12
[0263] .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-32 compounds, general building
blocks have been synthesized beforehand:
Building Block 1 (BB-1)
N'-[(5Z)-6-iodohex-5-en-1-yl]-N-methylethanediamide
[0264] Step 1:
[0265] PPh3 (140 g) and phthalimide (82.5 g) were suspended in dry
THF (500.0 mL) and cooled to 0.degree. 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.degree. C. for 1 h and then
at r.t. over night.
[0266] 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).
[0267] 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 (2.times.75 mL) and brine
(50 mL), dried over Na2SO4 and concentrated in vacuo. The residue
was filtered through a patch of SiO.sub.2 (eluent
CH.sub.2Cl.sub.2). 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.
[0268] The yellow oil of F2 was dissolved in EtOAc (400 mL) and
washed with 1 N NaOH (3.times.150 mL) and brine (50 mL). The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue was purified by column chromatography on
SiO.sub.2 (eluent CH.sub.2Cl.sub.2). 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.degree. 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.
[0269] Step 2:
[0270] 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 aluminum 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.
[0271] The reaction mixture was decanted from the formed
precipitate, diluted with water (400 mL) and extracted with EtOAc
(3.times.200 mL). The combined organic layers were washed with
water (100 mL), sat. Na.sub.2SO.sub.3 (3.times.100 mL) and brine
(100 mL), dried over Na.sub.2SO.sub.4 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.
[0272] Step 3:
[0273] 2-Methyl-2-butene (29.4 mL) was added dropwise to a
0.degree. C. cold solution of BH 3*SMe2 (2.00 M in THF, 64.4 mL)
and stirred at 0.degree. C. for 1 h and then at r.t. for 1 h. The
mixture was then added dropwise to a 0.degree. 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.degree. C., then HOAc (30.0 mL) was added dropwise, stirred
for 30 min at 0.degree. C. and then at r.t. over night. Control by
LC/MS showed product.
[0274] THF was removed in vacuo as far as possible. The residue was
then slowly poured into a solution of NaOH (15.0 g) in H2O (200 mL)
and extracted with CH.sub.2Cl.sub.2 (3.times.100 mL). The combined
organic layers were washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was used for further
transformation as such.
[0275] Step 4:
[0276]
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.
[0277] MeOH was removed in vacuo. The residue was suspended in
CH.sub.2Cl.sub.2 (300 mL). The solid was filtered off and and
washed with CH.sub.2Cl.sub.2 (2.times.100 mL). The combined
filtrates were then washed with water (2.times.100 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the crude
product as orange oil which was used for further transformation as
such.
[0278] Step 5:
[0279] Ethylchlorformylformiat (10.0 g) was dissolved in THF (50
mL) and cooled to 0.degree. C. Pyridine (7.70 mL) was added added
dropwise and the mixture stirred at 0.degree. C. for 30 min.
Methylamine (2.0 M in THF, 47.6 mL) was then added dropwise.
Stirring was continued at 0.degree. C. for 3 h. Control by TLC
(PE/EtOAc=1:3) showed product.
[0280] The precipitated salt was filtered off and the filtrate
evaporated. The residue was taken up in EtOAc (200 mL), washed with
1N HCl (2.times.50 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to give the product in sufficient purity as
brown oil.
[0281] Step 6:
[0282] (5Z)-6-iodohex-5-en-1-amine (11.15 g) was dissolved in EtOH
(200 mL). ethyl (methylcarbamoyl)formate (6.50 g) and NEt.sub.3
(8.26 mL) were added and the resulting mixture was stirred at
50.degree. C. for 24 h. Control by LC/MS showed incomplete
conversion. Additional (methylcarbamoyl)formate (1.00 g) and
NEt.sub.3 (4.00 mL) were added and stirring was continued at
50.degree. C. for 24 h. Control by LC/MS showed product.
[0283] EtOH was removed in vacuo. The residue was purified by
column chromatography on SiO.sub.2
(CH.sub.2Cl.sub.2.fwdarw.CH.sub.2Cl.sub.2/MeOH=50:1.fwdarw.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 icecold EtOAc
and dried in vacuo.
[0284] Yield: 10.3 g (67%) pale yellow solid.
Building Block 2 (BB-2)
N'-[4-(2-iodophenyl)butyl]-N-methylethanediamide
[0285] Step 1:
[0286] 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.degree. C.
Diisopropyl azodicarboxylate (75.1 mL) was then added dropwise over
a period of 20 min. The resulting mixture was stirred at 0.degree.
C. for 30 min and then at r.t. over night. Control by LC/MS showed
product.
[0287] 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.sub.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.
[0288] Step 2:
[0289] 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.degree. C. and the
resulting mixture was stirred at 0.degree. 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 PdCl.sub.2(PPh.sub.3).sub.2 (2.80
g) was added and the mixture heated to 50.degree. C. for 4 h.
Control by LC/MS showed product.
[0290] 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 Na.sub.2SO.sub.4. The residue was purified by column
chromatography on SiO.sub.2 (PE/EtOAc=6:4).
[0291] Step 3:
[0292]
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. H.sub.2SO.sub.4 (13.9 mL) were added and the resulting
suspension was cooled to 0.degree. C. A solution of NaNO.sub.2
(5.23 g) in water (50 mL) was added dropwise and the mixture
stirred at 0.degree. 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.
[0293] The reaction mixture was diluted with sat. Na.sub.2SO.sub.3
(200 mL) and extracted with EtOAc (3.times.200 mL). The combined
organic layers were washed with brine (150 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on SiO.sub.2 (PE/EtOAc=8:2).
[0294] Step 4:
[0295] 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 3
d. Control by LC/MS showed product.
[0296] MeOH was removed in vacuo. The residue was suspended in
CH.sub.2Cl.sub.2 (200 mL). The solid was filtered off and washed
with CH.sub.2Cl.sub.2 (100 mL). The combined filtrates were then
washed with water (2.times.100 mL). The combined aqueous layers
were reextracted with CH.sub.2Cl.sub.2 (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.
[0297] Step 5:
[0298] Ethylchlorformylformiat (10.0 g) was dissolved in THF (50
mL) and cooled to 0.degree. C. Pyridine (7.70 mL) was added added
dropwise and the mixture stirred at 0.degree. C. for 30 min.
Methylamine (2.0 M in THF, 47.6 mL) was then added dropwise.
Stirring was continued at 0.degree. C. for 3 h. Control by TLC
(PE/EtOAc=1:3) showed product.
[0299] The precipitated salt was filtered off and the filtrate
evaporated. The residue was taken up in EtOAc (200 mL), washed with
1N HCl (2.times.50 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to give the product in sufficient purity as
brown oil.
[0300] Step6:
[0301] 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.degree. C. for 18 h. Control by LC/MS showed
product.
[0302] The reaction mixture was cooled to r.t. and EtOH was removed
in vacuo. The residue was filtered through a patch of SiO2
(CH.sub.2Cl.sub.2/MeOH=98:2). Further purification by
recrystallization from EtOAc.
[0303] 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
[0304] Step 1
[0305] NaH (60% in mineral oil, 771 mg) was suspended in dry THF
(20.0 mL). The mixture was cooled to 0.degree. C., then
6-Hepten-1-ol (1.18 mL) was added. Stirring was continued at
0.degree. 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.degree. C. for 1.5 h. Control by TLC
(PE/EtOAc=1:1) showed product.
[0306] The reaction mixture was poured into 1N NaOH (50 mL) and
extracted with EtOAc (2.times.30 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 (3.times.30 mL). The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the product in
sufficient purity as colorless oil.
[0307] Step 2
[0308] 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 3 d. Control by TLC (PE/EtOAc=9:1) showed product.
[0309] 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 (2.times.100 mL) and then discarded. The combined PE
fractions were washed with 1N HCl (100 mL) and 1N NaOH (100 mL),
dried over Na.sub.2SO.sub.4 and concentrated in vacuo to afford the
product in sufficient purity as colorless liquid.
[0310] Step 3
[0311] Methyl 2-(hept-6-en-1-yloxy)acetate (2.88 g) was placed in
100 mL flask and cooled to 0.degree. C. under and Ar atmosphere.
9-BBN (0.5 M in THF, 38.7 mL) was then added dropwise and the
resulting mixture stirred at 0.degree. C. for 30 min and then at
r.t. for 2 h. A solution of Na.sub.2CO.sub.3 (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 PdCl.sub.2(PPh.sub.3).sub.2 (453 mg) were added
and the mixture heated to 50.degree. C. for 1.5 h. Control by LC/MS
showed product.
[0312] 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 SiO.sub.2 (PE/EtOAc 1:1).
[0313] Step 4
[0314] 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.
[0315] The reaction mixture was poured into 1N HCl (30 mL). The
precipitated product was filtered off, washed with water and dried
in vacuo.
[0316] Yield: 869 mg (86%) beige solid.
Building Block 6 (BB-6)
2-[3-(2-{4-[(Methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetic
acid
[0317] Step 1:
[0318] NaH (60% in mineral oil, 15.2 g) was suspended in dry THF
(250 mL). The mixture was cooled to 0.degree. C., then allyl
alcohol (11.8 mL) was added. Stirring was continued at 0.degree. 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.degree. C. for 3 h and stirred at r.t. over night.
[0319] The reaction mixture was poured into water (250 mL) and
extracted with EtOAc (2.times.100 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 (3.times.100 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.
[0320] Step 2:
[0321] 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.
[0322] 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 (2.times.100 mL). Control by TLC showed most of the product
remaining in the oily residue, which was thus washed with MTBE
(4.times.100 mL). The PE and MTBE layers were combined and washed
with 1N HCl (3.times.100 mL) and brine (50 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the product in
sufficient purity as pale yellow liquid.
[0323] Step 3:
[0324] Methyl 2-(prop-2-en-1-yloxy)acetate (1.30 g) was placed in a
100 mL flask and cooled to 0.degree. C. under and Ar atmosphere.
9-BBN (0.5 M in THF, 25.0 mL) was then added dropwise and the
resulting mixture was stirred at 0.degree. C. for 30 min and then
at r.t. for 2 h. A solution of Na.sub.2CO.sub.3 (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.sub.2(PPh.sub.3).sub.2 (292 mg) were added and the
mixture heated to 50.degree. 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.sub.2CO.sub.3 (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
PdCl.sub.2(PPh.sub.3).sub.2 (200 mg), the mixture was stirred at
50.degree. C. for 2 h. Control by LC/MS showed product.
[0325] 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
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on SiO2 (PE/EtOAc 3:7).
[0326] Step 4:
[0327] 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.
[0328] The reaction mixture was acidified with 6N HCl and extracted
with EtOAc (3.times.40 mL). The combined organic layers were washed
with brine (30 mL), dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue was purified by a short column on SiO2
(CH.sub.2Cl.sub.2/MeOH=9:1).
[0329] Yield: 1.56 g (80%) beige solid.
Building Block 8 (BB-8)
N'-[(5Z)-13-hydroxytridec-5-en-1-yl]-N-methylethanediamide
[0330] Step 1:
[0331] 6-Hepten-1-ol (3.00 g) and imidazole (3.57 g) were dissolved
in DMF (20.0 mL). TIPSCI (6.18 mL) was added and the resulting
mixture was stirred at 60.degree. C. for 6 h. Control by TLC
(PE/EtOAc=8:2) showed almost complete conversion.
[0332] The reaction mixture was diluted with water (100 mL) and
extracted with MTBE (3.times.40 mL). The combined organic layers
were washed with 1N HCl (2.times.50 mL) and brine (20 mL), dried
over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on SiO.sub.2 (PE/EtOAc=95:5).
[0333] Step 2:
[0334] (Hept-6-en-1-yloxy)tris(propan-2-yl)silane (1.57 g) was
placed in 100 mL flask and cooled to 0.degree. C. under and Ar
atmosphere. 9-BBN (0.5 M in THF, 14.5 mL) was then added dropwise
and the resulting mixture stirred at 0.degree. C. for 30 min and
then at r.t. for 2 h. A solution of Na.sub.2CO.sub.3 (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 PdCl.sub.2(PPh.sub.3).sub.2 (170 mg) were added and the mixture
heated to 50.degree. C. for 2 h. Control by LC/MS showed
product.
[0335] The reaction mixture was cooled to r.t. and the layers were
separated. The aqueous layer was extracted with EtOAc (2.times.50
mL). The combined organic layers were washed with brine (30 mL),
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was filtered through a patch of SiO.sub.2 (PE/EtOAc=4:6). The so
obtained crude product was used for further transformation as
such.
[0336] Step 3:
[0337] (hept-6-en-1-yloxy)tris(propan-2-yl)silane (2.20 g, crude
IK-0357/16) was dissolved in THF (50 mL) and cooled to 0.degree. C.
TBAF*3H2O (2.29 g) was added and the resulting mixture was stirred
at 0.degree. 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.
[0338] The reaction mixture was poured into water (100 mL) and
extracted with EtOAc (3.times.40 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.fwdarw.EtOAc).
[0339] Yield: 1.11 g (77%) beige solid.
Building Block 9 (BB-9)
N'-{4-[2-(3-hydroxypropyl)phenyl]butyl}-Nmethylethanediamide
[0340] Step 1:
[0341] 2-Propen-1-ol (3.00 g) and imidazole (7.03 g) were dissolved
in DMF (20.0 mL). TIPSCI (14.4 mL) was added and the resulting
mixture was stirred at 60.degree. 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
(3.times.40 mL). The combined organic layers were washed with 1N
HCl (2.times.50 mL) and brine (20 mL), dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by column
chromatography on SiO.sub.2 (PE/EtOAc=95:5).
[0342] Step 2:
[0343] (prop-2-en-1-yloxy)tris(propan-2-yl)silane (1.33 g) was
placed in a 100 mL flask and cooled to 0.degree. 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.degree. C. for
30 min and then at r.t. for 2 h.
[0344] A solution of Na2CO3 (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.sub.2(PPh.sub.3).sub.2 (146 mg) were added and the mixture
heated to 50.degree. 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
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was passed
through a short column of SiO.sub.2 (PE/EtOAc 1:1). The still crude
product was then used for further transformation as such.
[0345] Step 3:
[0346]
N-methyl-N'-{4-[2-(3-{[tris(propan-2-yl)silyl]oxy}propyl)phenyl]but-
yl}ethanediamide (1.87 g, crude IK-0357/17) was dissolved in THF
(50 mL) and cooled to 0.degree. C. TBAF*3H.sub.2O (1.97 g) was
added and the resulting mixture was stirred at 0.degree. 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 (3.times.40 mL). The combined organic layers
were washed with brine (20 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was passed through a short
column on SiO.sub.2(PE/EtOAc=1:1.fwdarw.EtOAc).
[0347] Yield: 911 mg (75%) beige solid.
Building Block 11 (BB-11)
N'-[(5Z)-13-(2-aminoethoxy)tridec-5-en-1-yl]-Nmethylethanediamide
[0348] Step 1:
[0349] NaH (60% in mineral oil, 7.71 g) was suspended in dry THF
(200 mL). The mixture was cooled to 0.degree. C., then
6-Hepten-1-ol (11.8 mL) was added. Stirring was continued at
0.degree. 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.degree. C. for 3 h. Control by TLC (PE/EtOAc=1:1)
showed product.
[0350] The reaction mixture was poured into 1N NaOH (300 mL) and
extracted with EtOAc (2.times.100 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 (3.times.100 mL). The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the product in
sufficient purity as pale brown oil.
[0351] Step 2:
[0352] 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 3 d. Control by TLC (PE/EtOAc=9:1) showed product.
[0353] 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 (2.times.100 mL) and then discarded. The combined PE
fractions were washed with 1N HCl (100 mL) and 1N NaOH (100 mL),
dried over Na.sub.2SO.sub.4 and concentrated in vacuo to afford the
product in sufficient purity as colorless liquid.
[0354] Step 3:
[0355] methyl 2-(hept-6-en-1-yloxy)acetate (5.00 g) was dissolved
in CH.sub.2Cl.sub.2 (100 mL) and cooled to 0.degree. C. DIBALH
(1.00 M in CH.sub.2Cl.sub.2, 61.7 mL) was added dropwise, and the
resulting mixture was stirred at 0.degree. C. for 30 min and then
at r.t. over night. Control by TLC (PE/EtOAc=8:2) showed complete
conversion.
[0356] The reaction mixture was cooled to 0.degree. C. and
carefully quenched with sat. aqueous Na.sub.2SO.sub.4. The mixture
was then diluted with CH.sub.2Cl.sub.2 (100 mL), stirred
vigourously for 20 min and then filtered through celite. The
filtercake was washed with CH.sub.2Cl.sub.2 several times. The
combined filtrates were concentrated in vacuo to afford the product
in sufficient purity as colorless liquid.
[0357] Step 4:
[0358] PPh.sub.3 (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.degree. C. Diisopropyl azodicarboxylate
(5.79 mL) was then added dropwise over a period of 20 min. The
resulting mixture was stirred at 0.degree. C. for 30 min and then
at r.t. over night.
[0359] 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 SiO.sub.2
(eluent PE/EtOAc=9:1) and evaporated. The residue was purified by
column chromatography on SiO.sub.2 (PE/EtOAc=8:2).
[0360] Step 5:
[0361]
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.degree. 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.degree. 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.sub.2(PPh.sub.3).sub.2 (226 mg) were added and the mixture
heated to 50.degree. C. for 1.5 h. Control by LC/MS showed
product.
[0362] The reaction mixture was cooled to r.t. and the layers were
separated. The aqueous layer was extracted with EtOAc (2.times.30
mL). The combined organic layers were washed with brine (30 mL),
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by column chromatography on SiO2 (PE/EtOAc=4:6).
[0363] Step 6:
[0364]
N'-[(5Z)-13-[2-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)ethoxy]tride-
c-5-en-1-yl]-N-methylethanediamide (2.36 g) was suspended in MeOH
(100 mL). Hydrazine hydrate (486 .mu.L) was added and the resulting
mixture was stirred at r.t. for 16 h. Control by LC/MS showed
product.
[0365] MeOH was removed in vacuo. The residue was suspended in
CH.sub.2Cl.sub.2/7N NH.sub.3 in MeOH=9:1 (100 mL) and filtered
through as patch of SiO.sub.2 and further eluated with
CH.sub.2Cl.sub.2/7N NH.sub.3 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
(CH.sub.2Cl.sub.2/7N NH3 in MeOH=9:1). The rest of the crude
material was used for further transformations as such.
[0366] 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
[0367] Step 1
[0368] NaH (60% in mineral oil, 7.71 g) was suspended in dry THF
(200 mL). The mixture was cooled to 0.degree. C., then
6-Hepten-1-ol (11.8 mL) was added. Stirring was continued at
0.degree. 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.degree. 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 (2.times.100 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 (3.times.100 mL). The combined organic layers were dried
over Na.sub.2SO.sub.4 and concentrated in vacuo to afford the
product in sufficient purity as pale brown oil.
[0369] Yield: 14.1 g (93%) pale brown oil
[0370] Step 2
[0371] A mixture of 2-(hept-6-en-1-yloxy)acetic acid (3.00 g) and
SOCl.sub.2 (15.00 mL) was heated to 70.degree. 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 (3.times.30 mL). The combined organic
layers were washed with sat. NaHCO3 (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
[0372] Step 3
[0373] 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.degree. C. A solution of POCl3 (1.54 mL) in CH2Cl2 (4 mL) was
slowly added. Stirring was then continued at 0.degree. C. for 15
min. Control by TLC (PE/EtOAc=8:2) showed product.
[0374] sat. NaHCO3 (5.00 mL) was added at 0.degree. 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
(3.times.20 mL). The combined organic layers were washed with sat.
NaHCO3 (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%.
[0375] Step 4
[0376] 2-(hept-6-en-1-yloxy)acetonitrile was placed in a 10 mL
flask and cooled to 0.degree. 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.degree. C. for 30 min and then at r.t. for
2 h. A solution of Na.sub.2CO.sub.3 (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 PdCl.sub.2(PPh.sub.3).sub.2 (19 mg) were added and the mixture
heated to 50.degree. 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%.
[0377] Step 5
[0378]
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.degree. C. overnight.
[0379] Water and ethyl acetate were added. The mixture was
acidified with HCl3N. The aqueous layer (acid pH) is then extracted
with ethyl acetate (.times.3), and the combined organic layer
washed with brine. The organic layer was dried over MgSO.sub.4,
filtered and solvent removed under vacuo. m=82 mg. Product was
purified by preparative TLC (DCM/MeOH 95/5)
[0380] Yield: 8 mg (10%), as white powder.
Compound 15 (Comp-15)
N'-(4-{2-[3-(carbamoylmethoxy)propyl]phenyl}butyl)-N-methylethanediamide
[0381] 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,
[0382] 5.35 ml) ammonia (0.4 M in THF) were added and the mixture
was stirred at rt over the weekend.
[0383] The mixture was poured into 50 ml water and extracted with
DCM (3.times.50 ml). The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated.
[0384] 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]tri-
dec-5-en-1-yl]ethanediamide
[0385] Step 1:
[0386] NaH (60% in mineral oil, 7.71 g) was suspended in dry THF
(200 mL). The mixture was cooled to 0.degree. C., then
6-Hepten-1-ol (11.8 mL) was added. Stirring was continued at
0.degree. 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.degree. 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 (2.times.100 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 (3.times.100 mL). The combined organic layers were dried
over Na2SO4 and concentrated in vacuo to afford the product in
sufficient purity as pale brown oil.
[0387] Step 2:
[0388] 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 3 d. 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 (2.times.100
mL) and then discarded. The combined PE fractions were washed with
1N HCl (100 mL) and 1N NaOH (100 mL), dried over Na.sub.2SO.sub.4
and concentrated in vacuo to afford the product in sufficient
purity as colorless liquid.
[0389] Step 3:
[0390] 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.degree. C.
for 4.5 h (.fwdarw.clear solution).
[0391] .fwdarw.LC/MS: GH-0513/1-1
[0392] .fwdarw.TLC (EA/PE 1:1): Complete consumption of starting
material
[0393] The mixture was evaporated to dryness
[0394] Step 4:
[0395] 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.
[0396] Step 5:
[0397] The oily residue was dissolved in 10 ml 2M NaOH and heated
to reflux for 2 h..fwdarw.LC/MS: GH-0515/1-2: Complete consumption
of intermediate 1. The mixture was acidified using conc. HCl and
extracted with EA (3.times.20 ml). The combined organic layers were
dried over Na.sub.2SO.sub.4 and concentrated to dryness. The crude
solid was recrystallized from ACN.
[0398] Step 6:
[0399] 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) Na.sub.2CO.sub.3 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 .mu.mol) PdCl 2(PPh3)2 were
added and the mixture was heated to 50.degree. C. for 4 h (a yellow
biphasic mixture).
[0400] .fwdarw.LC/MS: GH-0516/1-1: Product was detected
[0401] 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 a 9:1, Rf of possible
product: 0.62). Recrystallization from CAN.
[0402] Yield: 51 mg (0.13 mmol, 45%).
Compound 17 (Comp-17)
N-methyl-N'-[(5Z)-13-[(phenylcarbamoyl)methoxy]tridec-5-en-1-yl]ethanediam-
ide
[0403]
2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic
acid (BB-4, 50.0 mg, 140.3 .mu.mol), Aniline (26 .mu.l, 280.5
.mu.mol), HBTU (53.4 mg, 140.3 .mu.mol) and DMAP (1.7 mg, 14.0
.mu.mol) were placed in a G16 vial. DMF (2.00 ml) and NEt 3 (78.0
.mu.l, 561.1 .mu.mol) were added and the resulting mixture was
stirred at r.t. for 16 h. Control by LC/MS showed product.
[0404] The reaction mixture was diluted with water (20 ml) and was
extracted with Et 20 (3.times.20 ml). The combined organic layers
were washed with sat. NaHCO.sub.3 (20 ml) and brine (10 ml), dried
over Na.sub.2SO.sub.4 and concentrated in vacuo. The product was
lyophilized.
[0405] 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
[0406]
2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic
acid (BB-4, 50.0 mg, 140.3 .mu.mol), 4-Aminotetrahydropyran (29
.mu.l, 280.5 .mu.mol), HBTU (53.4 mg, 140.3 .mu.mol) and DMAP (1.7
mg, 14.0 .mu.mol) were placed in a G16 vial. DMF (2.00 ml) and NEt3
(78.0 .mu.l, 561.1 .mu.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 (3.times.20 ml). The combined organic layers were washed
with sat. NaHCO3 (20 ml) and brine (10 ml), dried over Na2SO4 and
concentrated in vacuo. The product was lyophilized.
[0407] 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
[0408]
2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic
acid (BB-4, 50.0 mg, 140.3 .mu.mol), 1,3-Oxazol-2-amine (24 mg,
280.5 .mu.mol), HBTU (53.4 mg, 140.3 .mu.mol) and DMAP (1.7 mg,
14.0 .mu.mol) were placed in a G16 vial. DMF (2.00 ml) and
NEt.sub.3 (78.0 .mu.l, 561.1 .mu.mol) were added and the resulting
mixture was stirred at r.t. for 16 h. Control by LC/MS showed
product.
[0409] The reaction mixture was diluted with water (20 ml) and was
extracted with Et.sub.2O (3.times.20 ml). The combined organic
layers were washed with sat. NaHCO3 (20 ml) and brine (10 ml),
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by preparative TLC (DCM/MeOH=95:5).
[0410] Yield: 11 mg (19%), white solid.
Compound 20 (Comp-20)
N'-[(5Z)-13-{[(4-methoxyphenyl)carbamoyl]methoxy}tridec-5-en-1-yl]-Nmethyl-
ethanediamide
[0411]
2-{[(8Z)-13-[(methylcarbamoyl)formamido]tridec-8-en-1-yl]oxy}acetic
acid (BB-4, 50.0 mg, 140.3 .mu.mol), p-Anisidine (35 mg, 280.5
.mu.mol), HBTU (53.4 mg, 140.3 .mu.mol) and DMAP (1.7 mg, 14.0
.mu.mol) were placed in a G16 vial. DMF (2.00 ml) and NEt.sub.3
(78.0 .mu.l, 561.1 .mu.mol) were added and the resulting mixture
was stirred at r.t. for 16 h. Control by LC/MS showed product.
[0412] The reaction mixture was diluted with water (20 ml) and was
extracted with Et.sub.2O (3.times.20 ml). The combined organic
layers were washed with sat. NaHCO3 (20 ml) and brine (10 ml),
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by preparative TLC (DCM/MeOH=98:2).
[0413] Yield: 18 mg (28%), beige solid.
Compound 21 (Comp-21)
N-Methyl-N'-[4-(2-{3-[(phenylcarbamoyl)methoxy]propyl}phenyl)butyl]
ethanediamide
[0414] 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
.mu.l) aniline were dissolved in 3 ml DCM and stirred at rt of the
weekend (clear solution).
[0415] The mixture was evaporated to dryness and purified via pTLC
(1 mm, DCM/MeOH 20:1, Rf of possible product: 0.54).
[0416] 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
[0417] 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
.mu.l) Pyrrolidine were dissolved in 3 ml DCM and stirred at rt for
1.5 h (clear solution).
[0418] The mixture was evaporated to dryness and purified via pTLC
(1 mm, DCM/MeOH 10:1, Rf of possible product: 0.46).
[0419] 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
[0420] 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 .mu.l) mor pholine were dissolved in 3 ml DCM and
stirred at rt over the weekend (clear solution).
[0421] The mixture was evaporated to dryness and purified via pTLC
(1 mm, DCM/MeOH 10:1, Rf of possible product: 0.48).
[0422] Yield: 34 mg (58%), white solid.
Compound 24 (Comp-24)
4-{2-[3-(2-{4-[(methylcarbamoyl)formamido]butyl}phenyl)propoxy]acetamido}b-
enzoic acid
[0423] STEP 1:
[0424] 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).
[0425] The mixture was evaporated to dryness and purified via pTLC
(1 mm, EA/PE 4:1, R f of possible product: 0.31)
[0426] STEP 2:
[0427] To a solution of 48 mg (0.10 mmol)
N-Methyl-N'-{4-[2-(3-{[(pyridin-2-yl)carbamoyl]methoxy}
[0428] 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).
[0429] The mixture was poured into 1 N HCl solution (10 ml) and
extracted with DCM (3.times.20 ml). The combined organic layers
were dried over Na.sub.2SO.sub.4 and concentrated to dryness. The
crude product was purified via pTLC (DCM/MeOH/FA 100:10:1, Rf of
possible product: 0.43).
[0430] Yield: 10 mg (21%), white solid.
Compound 25 (Comp-25)
N'-[(5Z)-13-[2-(4-hydroxy-2-oxo-2,5-dihydrofuran-3-yl)-2-oxoethoxy]tridec--
5-en-1-yl]-Nmethylethanediamide
[0431]
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.
CH.sub.2Cl.sub.2 (3.00 mL) was added and the resulting mixture was
stirred at r.t. for 18 h. Control by LC/MS showed product.
[0432] The reaction mixture was diluted with water (30 mL) and
extracted with CH.sub.2Cl.sub.2 (3.times.20 mL). The combined
organic layers were washed with brine (10 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by preparative TLC (CH.sub.2Cl.sub.2/MeOH=9:1).
[0433] Yield: 20 mg (33%), beige solid.
Compound 26 (Comp-26)
N'-[4-(2-(3-[2-(hydroxymethyl)
phenoxyl]propyl)phenyl)butyl]-N-methylethanediamide
[0434] Step 1:
[0435] Salicylaldehyde (2.00 g) and imidazole (2.79 g) were
dissolved in DMF (20.0 mL). TIPSCI (5.96 mL) was added and the
resulting mixture was stirred at 60.degree. C. for 2 d. Control by
TLC (PE/EtOAc=95:5) and LC/MS showed incomplete conversion.
Additional TIPSCI (2.00 mL) was added and stirring was continued at
60.degree. C. for 3 d. 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 (3.times.40 mL). The
combined organic layers were washed with 1N NaOH (30 mL) and brine
(20 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by column chromatography on SiO.sub.2
(PE/EtOAc=95:5).
[0436] Yield: 3.54 g (78%) pale yellow liquid
[0437] Step 2:
[0438] 2-{[tris(propan-2-yl)silyl]oxy}benzaldehyde (3.54 g) was
dissolved in EtOH (30.0 mL) and cooled to 0.degree. C. NaBH4 (481
mg) was added and the resulting mixture was stirred at 0.degree. 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 (3.times.40 mL). The
combined organic layers were washed with brine (30 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on SiO.sub.2 (PE/EtOAc=8:2).
[0439] Yield: 2.73 g (77%) yellow oil
[0440] Step 3:
[0441] (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
.mu.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 CH.sub.2Cl.sub.2
(3.times.30 mL). The combined organic layers were washed with brine
(20 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
so obtained crude product was used for further transformation as
such.
[0442] Yield: 480 mg (crude) yellow oil
[0443] Step 4:
[0444]
{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.degree. 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.degree. 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.degree. 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 SiO.sub.2 (PE/EtOAc 1:1). The
still crude product was then used for further transformation as
such.
[0445] Yield: 235 mg (crude) yellow oil.
[0446] Step 5:
[0447]
N-methyl-N'-[4-(2-{3-[(2-{[tris(propan-2-yl)silyl]oxy}phenyl)methox-
y]propyl}phenyl)butyl]ethanediamide (154 mg, crude IK-0357/19) was
dissolved in THF (5.00 mL). TBAF*3H2O (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 (3.times.10 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 (CH.sub.2Cl.sub.2/MeOH=95:5).
[0448] Yield: 75 mg (66%) white solid.
Compound 27 (Comp-27)
N'-[(5Z)-13-(2-hydroxyphenoxy)tridec-5-en-1-yl]-Nmethylethanediamide
[0449] Step 1:
[0450] N'-[(5Z)-13-hydroxytridec-5-en-1-yl]-N-methylethanediamide
(BB-8, 400 mg) was suspended in CH.sub.2Cl.sub.2 (20 mL). PPh3 (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.
[0451] The reaction mixture was concentrated in vacuo and the
residue was purified by preparative TLC (PE/EtOAc=1:1).
[0452] Step 2:
[0453] N'-[(5Z)-13-bromotridec-5-en-1-yl]-N-methylethanediamide (50
mg), pyrocatechol (76.2 mg) and K.sub.2CO.sub.3 (57.4 mg) were
placed in a G16 vial. DMF (3.00 mL) was added and the resulting
mixture was stirred at 60.degree. C. for 2.5 h. Control by LC/MS
showed product.
[0454] The reaction mixture was diluted with water (40 mL) and
extracted with MTBE (3.times.20 mL). The combined organic layers
were washed with water (20 mL) and brine (10 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by preparative TLC (CH.sub.2Cl.sub.2/MeOH=95:5).
[0455] Yield: 43 mg (80%), white solid.
Compound 28 (Comp-28)
N-Methyl-N'-[4-(2-{3-[2-(morpholin-4-yl)-2-oxoethoxy]propyl}phenyl)butyl]
ethanediamide
[0456] 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 .mu.l) morpholine were dissolved in 3 ml DCM and stirred
at rt over the weekend (clear solution).
[0457] The mixture was evaporated to dryness and purified via pTLC
(1 mm, DCM/MeOH 10:1, Rf of possible product: 0.48).
[0458] Yield: 34 mg (58%), white solid.
Compound 29 (Comp-29)
N'-(4-{2-[3-(3-hydroxyphenoxy)propyl]phenyl}butyl)-Nmethylethanediamide
[0459] Step 1:
[0460]
N'-{4-[2-(3-hydroxypropyl)phenyl]butyl}-N-methylethanediamide
(BB-9, 400 mg) was suspended in CH.sub.2Cl.sub.2 (20 mL). PPh.sub.3
(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).
[0461] Step 2:
[0462] N'-{4-[2-(3-bromopropyl)phenyl]butyl}-N-methylethanediamide
(50 mg), 1,3-Benzenediol (77.5 mg) and K.sub.2CO.sub.3 (58.4 mg)
were placed in a G16 vial. DMF (3.00 mL) was added and the
resulting mixture was stirred at 60.degree. C. for 1 h. Control by
LC/MS showed product.
[0463] The reaction mixture was diluted with water (40 mL) and
extracted with MTBE (3.times.20 mL). The combined organic layers
were washed with water (20 mL) and brine (10 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by preparative TLC (CH.sub.2Cl.sub.2/MeOH=95:5).
[0464] Yield: 40 mg (74%), white solid.
Compound 30 (Comp-30)
N'-[(5Z)-13-(4-hydroxyphenoxy)tridec-5-en-1-yl]-Nmethylethanediamide
[0465] Step 1:
[0466] N'-[(5Z)-13-hydroxytridec-5-en-1-yl]-N-methylethanediamide
(BB-8, 400 mg) was suspended in CH2Cl2 (20 mL). PPh3 (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.
[0467] The reaction mixture was concentrated in vacuo and the
residue was purified by preparative TLC (PE/EtOAc=1:1).
[0468] Step 2:
[0469] N'-[(5Z)-13-bromotridec-5-en-1-yl]-N-methylethanediamide (50
mg), hydroquinone (76.2 mg) and K.sub.2CO.sub.3 (57.4 mg) were
placed in a G16 vial. DMF (3.00 mL) was added and the resulting
mixture was stirred at 60.degree. C. for 2.5 h. Control by LC/MS
showed product.
[0470] The reaction mixture was diluted with water (40 mL) and
extracted with MTBE (3.times.20 mL). The combined organic layers
were washed with water (20 mL) and brine (10 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by preparative TLC (CH.sub.2Cl.sub.2/MeOH=95:5).
[0471] Yield: 43 mg (80%), white solid
Compound 31 (Comp-31)
N'-(4-{2-[3-(4-hydroxyphenoxy)propyl]phenyl}butyl)-Nmethylethanediamide
[0472] Step 1:
[0473]
N'-{4-[2-(3-hydroxypropyl)phenyl]butyl}-N-methylethanediamide
(BB-9, 400 mg) was suspended in CH2Cl2 (20 mL). PPh3 (610 mg) and
CBr.sub.4 (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.
[0474] The reaction mixture was concentrated in vacuo and the
residue was purified by preparative TLC (PE/EtOAc=1:1).
[0475] Step 2:
[0476] N'-{4-[2-(3-bromopropyl)phenyl]butyl}-N-methylethanediamide
(50 mg), Hydroquinone (77.5 mg) and K.sub.2CO.sub.3 (58.4 mg) were
placed in a G16 vial. DMF (3.00 mL) was added and the resulting
mixture was stirred at 60.degree. C. for 1 h. Control by LC/MS
showed product.
[0477] The reaction mixture was diluted with water (40 mL) and
extracted with MTBE (3.times.20 mL). The combined organic layers
were washed with water (20 mL) and brine (10 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by preparative TLC (CH.sub.2Cl.sub.2/MeOH=95:5).
[0478] Yield: 39 mg (72%), white solid
Compound 3'' (Comp-3'')
N'-[4-(2-{3-[(4-hydroxyphenyl)methoxy]propyl}phenyl)butyl]-N-methylethaned-
iamide
[0479] Step 1
[0480] 4-Hydroxybenzaldehyde (2.00 g) and imidazole (2.79 g) were
dissolved in DMF (20.0 mL). TIPSCI (5.96 mL) was added and the
resulting mixture was stirred at 60.degree. C. for 2 d.
[0481] Control by TLC (PE/EtOAc=95:5) and LC/MS showed complete
conversion.
[0482] The reaction mixture was diluted with water (100 mL) and
extracted with MTBE (3.times.40 mL). The combined organic layers
were washed with 1N NaOH (30 mL) and brine (20 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on SiO.sub.2 (PE/EtOAc=95:5).
Yield: 3.94 g (86%) pale yellow oil
[0483] Step 2
[0484] 4-{[tris(propan-2-yl)silyl]oxy}benzaldehyde (3.94 g) was
dissolved in EtOH (30.0 mL) and cooled to 0.degree. C. NaBH4 (535
mg) was added and the resulting mixture was stirred at 0.degree. C.
for 30 min and then at r.t. for 18 h.
[0485] Control by TLC (PE/EtOAc=8:2) and LC/MS showed product.
[0486] The reaction mixture was diluted with water (100 mL) and
extracted with EtOAc (3.times.40 mL). The combined organic layers
were washed with brine (30 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by column
chromatography on SiO.sub.2 (PE/EtOAc=8:2).
[0487] Step 3
[0488] (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 .mu.L) was then added and the resulting mixture was stirred at
r.t. for 2.5 h. Control by LC/MS showed complete conversion.
[0489] The reaction mixture was poured into water (30 mL) and
extracted with EtOAc (3.times.10 mL). The combined organic layers
were washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was used for further
transformation as such.
[0490] Yield: 372 mg (crude) yellow oil
[0491] Step 4
[0492]
{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.degree. 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.degree. C. for
30 min and then at r.t. for 2 h.
[0493] A solution of Na.sub.2CO.sub.3 (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
PdCl.sub.2(PPh.sub.3).sub.2 (9.7 mg) were added and the mixture
heated to 50.degree. 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.
[0494] Yield: 248 mg (crude) yellow oil.
[0495] Step 5
[0496]
N-methyl-N'-[4-(2-{3-[(4-{[tris(propan-2-yl)silyl]oxy}phenyl)methox-
y]propyl}phenyl)butyl]ethane-diamide (154 mg, crude IK-0357/20) was
dissolved in THF (5.00 mL). TBAF*3H2O (131 mg) was added and the
resulting mixture was stirred at r.t. for 30 min. Control by LC/MS
showed complete conversion.
[0497] The reaction mixture was poured into water (40 mL) and
extracted with EtOAc (3.times.20 mL). The combined organic layers
were washed with brine (10 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by preparative TLC
(CH.sub.2Cl.sub.2/MeOH=95:5).
[0498] Yield: 76 mg (68%), white solid.
TABLE-US-00019 TABLE 1 Calculated exact mass of Comp-14 to Comp-32
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
4-{2-[3-(2-{4- Comp-24 469.22129 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-25 438.2366 439.243883 en-1-yl]-N-methylethanediamide
N'-[4-(2-{3-[2-(hydroxymethyl)phenoxy]propyl}phenyl)butyl]-N-
Comp-26 398.22056 399.227838 methylethanediamide
N'-[(5Z)-13-(2-hydroxyphenoxy)tridec-5-en-1-yl]-N-methylethanediamide
Comp-27 390.25186 391.259138
N'-[(5Z)-13-(3-hydroxyphenoxy)tridec-5-en-1-yl]-N-methylethanediamide
Comp-28 390.25186 391.259138
N'-(4-{2-[3-(3-hydroxyphenoxy)propyl]phenyl}butyl)-N-methylethanediamide
Comp-29 384.20491 385.212188
N'-[(5Z)-13-(4-hydroxyphenoxy)tridec-5-en-1-yl]-N-methylethanediamide
Comp-30 390.25186 391.259138
N'-(4-{2-[3-(4-hydroxyphenoxy)propyl]phenyl}butyl)-N-methylethanediamide
Comp-31 384.20491 385.212188
N'-[4-(2-{3-[(4-hydroxyphenyl)methoxy]propyl}phenyl)butyl]-N-
Comp-32 398.22056 399.227838 methylethanediamide
Example 2
Efficacy of Compound 1 in a Laser Induced Choroidal
Neovascularization Model in Rat Evaluated by Measuring Vascular
Leakage Using Fluorescence Angiography
[0499] The laser induced CNV model in rat is a widely used model to
prove the therapeutic efficacy of drugs for treatment of several
ocular diseases characterized by angiogenesis and inflammation in
the eye such as wet age-related macular degeneration (AMD). In this
model a focused laser burn in the layer of Bruch's membrane causes
a disruption of this membrane, a local injury, followed by
inflammation and growth of new blood vessels. These newly formed
blood vessels are typically leaky. This leakage is measured by
extravasation of a fluorescent dye in the back of the eye
(Fluorescein) and is a well-accepted marker for vascular leakage
which is proportionally to the amount of newly grown blood vessels.
This method is also the state of the art procedure in the clinical
practice in man.
[0500] All standard operating procedures and protocols described in
this study plan have been reviewed by Ethics Committee. All animals
have been treated according to the Directive 2010/63/UE European
Convention for the Protection of Vertebrate Animals used for
Experimental and Other Scientific Purposes and to the Association
for Research in Vision and Ophthalmology (ARVO) Statement for the
Use of Animals in Ophthalmic and Vision Research.
[0501] Animals: Brown Norway rats at the age of 8-10 weeks at the
day of induction.
[0502] Induction of neovascularization: Animals have been
anesthetized by an intramuscular injection of a mix xylazine and
ketamine. Right eye pupils have been dilated by instillation of one
drop of 0.5% tropicamide before handling. Six burns on anesthetized
animals have been created in the right eye by applying 170-180 mW
of 532-nm laser light (Viridis laser, Quantel, France) on 75-.mu.m
spots around the optic nerve, between the main retinal vessel
branches, for 0.1 s, through the slit lamp and a contact lens.
Production of a bubble at the time of laser application confirmed
the rupture of Bruch's membrane.
[0503] Treatment: For vehicle control group on Day 0, just after
induction of neovascularization, vehicle was intravitreally
injected (5 .mu.L) in right eyes under an operating microscope
using a 30-G-needle mounted on a 100 .mu.L Hamilton syringe. The
administrations have been made on anesthetized animals (same
anesthesia than for neovascularization induction). In the first
treatment group Compound 1, which has a structure as indicated in
formula (VI) was intraperitoneally administrated (100 .mu.L/100 g),
once daily from Day 0 (3 hours.+-.15 min prior induction) to Day
23. In another treatment group Compound 1 was orally administrated
via gavage (1 mL/100 g), twice daily from Day 0 (3 hours.+-.15 min
prior induction) to Day 22 (q7 h.+-.30 min).
[0504] Fluorescein angiography: Fluorescein angiography has been
performed in the lasered right eyes on Days 14 and 21 using
Heidelberg Retinal Angiograph (HRA). After anesthesia (see above),
250 .mu.L/100 g body weight of a 10% sodium fluorescein will be
injected subcutaneously, and fluorescence photos will be recorded
10 minutes after dye injection. For the evaluation by fluorescein
angiography the leakage of fluorescein will be evaluated in the
angiograms by two independent examiners masked to the study groups
and graded as follows: Score 0: no leakage; Score 1: slightly
stained; Score 2: moderately stained; Score 3: strongly
stained.
[0505] In-life phase termination: At the end of the study on Day
23, all animals will be anesthetized (see above) then euthanized
using an overdosed intracardiac injection of pentobarbital. This
method is among the recommended methods by the European
Authorities.
[0506] Statistical analysis: The statistical analyses have been
performed using the software Prism. A Kruskal-Wallis analysis has
been performed and the drug effect has been assessed using the
Dunett's test for multiple comparisons; each treated group has been
compared to that of the vehicle at each time-point. The p values
lower than 0.05 will be significant.
[0507] Results are shown in FIG. 1.
Example 3
Comp-02 Distribution after Oral, Topical or Intravitreal
Administration in Different Compartments of the Rabbit Eye
[0508] This example shows that highest ocular exposures of Comp-02
in the posterior eye are reached after intravitreal injection.
Materials and Methods
[0509] Study design: To gain insight into the distribution
behaviour of synthetic 17,18-EEQ-agonists, ocular pharmacokinetic
studies were performed in pigmented HY79b rabbits. Briefly, 15 male
rabbits per administration route were anesthetized at the
time-points for sampling indicated in tables 2-4 by an
intramuscular injection of a mixed solution of xylazine and
ketamine. Blood collection was performed by cardiac puncture in
K.sub.3EDTA tubes and centrifuged at 2000 g, 10 min at 4.degree. C.
Approximately 3 mL of plasma were sampled, put in plastic tubes,
snap frozen in liquid nitrogen and stored at -80.degree. C. until
assay. Then animals were euthanized by intracardiac injection of
overdosed pentobarbital. This method is one of the recommended
methods for euthanasia by the European authorities (French decree
No. 2013-118, dated Feb. 1, 2013 publishing the European directive
2010/63/UE. J.Offic.Rp.Fr. 2013; Text 24 out of 130). Immediately
after euthanasia, samples from both eyes were quickly and carefully
taken, put in plastic tubes, weighed, snap frozen in liquid
nitrogen and stored at -80.degree. C. until assay. Content of
Comp-02 was determined in ocular samples and plasma following an
established RRLCMS/MS method. The results are presented in Tables
5-7 and are expressed as mean values for the different
pharmacokinetic parameters. Chromatogram integration was done using
MassHunter software. Apparent Cmax and Tmax, T1/2, AUC1-48 h were
calculated by Excel.RTM. software. Concentrations were expressed in
ng/g or ng/mL of tissue/eye compartment.
Results
[0510] After oral administration Comp-02 was below lower limit of
detection (0.1 ng/10 .mu.l injected) in both aqueous humor (except
for one out of 15 animals) and retina (except for two out of 15
animals) samples. However, oral administration of Comp-02 reached
effective dose level (246 ng/g) 1 h after administration of 2 mg/kg
(0.2 mg/mL) in choroid tissue as well as considerable plasma level
(1213 ng/mL, 1 h after administration). Corresponding to the
maximal level considerable exposures were only seen in choroid and
plasma (AUC.sub.1-48hrs (ng*h/mL): 1387 and 6331).
[0511] For topical administration it was found, that the Comp-02
formulation was macroscopically very well tolerated, as checked by
ophthalmoscopy. Table 6 shows, besides generally very low level,
that highest dose level were seen in aqueous humor (88 ng/mL 1 h
post-administration), followed by the choroid (79 ng/g 1 h
post-administration). Both in the retina and in plasma only low
level were reached (24 ng/g 1 h post-administration and 31 ng/mL
0.5 h post-administration, respectively). AUC.sub.1-48 hrs values
were low after topical administration in all tested ocular tissues
(AUC.sub.1-48 hrs (ng*h/mL): 324 for aqueous humor, 117 for choroid
and 21 for retina) as well as in plasma (262 ng*h/mL).
[0512] Also for intravitreal injection it was found, that the
Comp-02 formulation was macroscopically very well tolerated, as
checked by ocular examination with a slit lamp. Table 6 shows, that
considerable Comp-02 level were found in all tissues of the eye and
in plasma. Highest level were found in the choroid (14957 ng/g 1 h
post-injection) followed by retina and vitreous (10052 ng/g and
6647 ng/mL 1 h post-injection). Exposures were >55000
(AUC.sub.1-48 hrs (ng*h/mL): 55401 for vitreous, 63283 for choroid
and 79866 for retina) except for plasma (287 ng*h/mL)).
[0513] In summary, the highest ocular exposures of Comp-02 in the
posterior eye were reached after intravitreal injection compared to
oral and topical administration.
TABLE-US-00020 TABLE 2 Study design for oral administration
Time-points Animal Group Formulation Administration for sampling
number 1 Comp-02 in T0 h: Single oral 1 h .+-. 6 min 1, 2, 3 2
isotonic salt administration 4 h .+-. 24 min 4, 5, 6 3 solution 2
mg/kg, 0.2 mg/mL 8 h .+-. 48 min 7, 8, 9 4 24 h .+-. 144 min 10,
11, 12 5 48 h .+-. 288 min 13, 14, 15
TABLE-US-00021 TABLE 3 Study design for topical administration
Time-points Animal Group Formulation Administration for sampling
number 1 Comp-02 in T0 h: Single 30 min .+-. 3 min 1, 2, 3 2
isotonic salt instillation 1 h .+-. 6 min 4, 5, 6 3 solution 50
.mu.l (1 mg/ 2 h .+-. 12 min 7, 8, 9 4 mL) in both 4 h .+-. 24 min
10, 11, 12 5 eyes 24 h .+-. 144 min 13, 14, 15
TABLE-US-00022 TABLE 4 Study design for intravitreal administration
Time-points Animal Group Formulation Administration for sampling
number 1 Comp-02 in T0 h: Single vitreal 1 h .+-. 6 min 1, 2, 3 2
isotonic salt injection of 50 .mu.l 2 h .+-. 12 min 4, 5, 6 3
solution (1 mg/mL) in both 8 h .+-. 48 min 7, 8, 9 4 eyes 24 h .+-.
144 min 10, 11, 12 5 48 h .+-. 288 min 13, 14, 15
TABLE-US-00023 TABLE 5 Pharmacokinetic parameters in ocular tissue
and plasma after oral administration Aqueous humor Retina Choroid
Plasma C.sub.max 8 15 246 1213 T.sub.max 1 4 1 1 t.sub.1/2 NA NA 9
6 AUC.sub.1-48 hours 12 64 1387 6331 Cmax = the highest mean value
measured (ng/g or mL of tissue) Tmax = time-point when the highest
mean value is measured (hours) AUC time 1 to time 2 = area under
the curve in ng/g or mL of tissue t1/2 = ln(2)/(-a) with a = slope
of ln (concentration) = f(t) t1/2 = calculated from the Tmax value
NA = Not applicable, insufficient data available
TABLE-US-00024 TABLE 6 Pharmacokinetic parameters in ocular tissue
and plasma after topical administration Aqueous humor Retina
Choroid Plasma C.sub.max 88 24 79 31 T.sub.max 1 1 1 0.5 t.sub.1/2
1 NA 0.7 5.4 AUC.sub.1-48 hours 324 21 117 262 Cmax = the highest
mean value measured (ng/g or mL of tissue) Tmax = time-point when
the highest mean value is measured (hours) AUC time 1 to time 2 =
area under the curve in ng/g or mL of tissue t1/2 = ln(2)/(-a) with
a = slope of ln (concentration) = f(t) t1/2 = calculated from the
Tmax value NA = Not applicable, insufficient data available
TABLE-US-00025 TABLE 7 Pharmacokinetic parameters in ocular tissue
and plasma after intravitreal injection Vitreous Retina Choroid
Plasma C.sub.max 6647 10052 14957 19 T.sub.max 1 1 1 2 t.sub.1/2
4.6 2.6 2.4 4.2 AUC.sub.1-48 hours 55401 79866 63283 287 Cmax = the
highest mean value measured (ng/g or mL of tissue) Tmax =
time-point when the highest mean value is measured (hours) AUC time
1 to time 2 = area under the curve in ng/g or mL of tissue t1/2 =
ln(2)/(-a) with a = slope of ln (concentration) = f(t) t1/2 =
calculated from the Tmax value NA = Not applicable, insufficient
data available
Example 4
Anti-Inflammatory Effect of a Metabolically Robust Analog of
17,18-EEQ (Comp-02) on HL-1 Cardiomyocytes
Materials and Methods
[0514] In order to investigate the anti-inflammatory potential of
compounds being part of the invention in vitro, a cardiomyocyte
cell line was used (mouse derived immortalized cardiomyocytes, HL-1
cells). Cells were either treated with vehicle (0.01% ethanol) or
different concentrations of test compound (Comp-02: c.sub.E=10 nM,
100 nM or 1 .mu.M). Simultaneously, the cells were challenged with
1 .mu.g/mL lipopolysaccharide (LPS). After 24 h of incubation, the
cells were processed to measure viability (FIG. 2) and release of
the pro-inflammatory cytokine TNF alpha (FIG. 3).
Results
[0515] The results are presented in FIG. 2 and FIG. 3. The
inflammatory stimulus LPS leads to a significant reduction in cell
viability. This cytotoxic effect was dose-dependently reversed by
Comp-02, FIG. 2. Moreover, LPS-incubation significantly induced the
production of the pro-inflammatory cytokine TNF alpha (FIG. 3).
Comp-02 alone did not have an influence on TNF alpha production.
LPS-induced TNF alpha release from the HL-1 cells was significantly
and dose-dependently reduced by Comp-02 (FIG. 3).
Example 5
Inhibitory Effect of a Metabolically Robust Analog of 17,18-EEQ
(Comp-02) on Renal and Cardiac Inflammation in a Rat Model of
Severe Hypertension and End-Organ Damage
Materials and Methods
[0516] The aim of this study was to assess and compare the effect
of continuous oral treatment of a metabolically robust analog of
17,18-EEQ (Comp-02) or vehicle (isotonic salt solution) on
physiological parameters, clinical chemistry and end-organ damage
in male double transgenic rats (dTGR). Non-treated Sprague-Dawley
rats (SD) of same genetic background were used as control. Double
transgenic rats, overexpressing the human renin and angiotensinogen
genes, develop severe hypertension and are a model for Ang-II
induced end-organ damage with cardiac hypertrophy and kidney
failure. Moreover, the dTGR model features severe atrial
hypertension, cardiac hypertrophy and nephrosclerosis with marked
fibrosis and inflammation.
[0517] Before start of the experiment the animals were randomly
assigned to either compound or vehicle treatment. Prior to the
experiment chow was changed to an omega-6 rich chow for all
treatment groups as well as the SD-control animals. dTGR compound
treated animals were dosed twice daily with 1.33 mg/kg for 21 days
by gavage. Comp-02 was prepared in 61.6 ppm Na2CO3 and 0.9% NaCl
ready to use solution for application, while dTGR animals were
dosed twice daily with 0.9% NaCl ready to use solution for vehicle
control. The application volume was 5.0 mL/kg b.w. After 21 days of
treatment, at the age of seven weeks, animals were sacrificed
through heart removal under pentobarbital anaesthesia (20 mg/kg
b.w., heparin 500 I.E./mL) and blood and organs were harvested.
Heart and kidney were removed and weighed. Heart and kidney were
divided and snap frozen in liquid nitrogen for molecular biology
analyses, frozen in -40.degree. C. isopentane and stored at
-80.degree. C. for cryoslides. Frozen kidneys and hearts embedded
in TissueTek were cryosectioned to 5 .mu.m thickness. Cryo sections
were fixed with ice-cold acetone, washed with PBS and blocked with
normal donkey serum at room temperature. Afterwards sections were
incubated in a dark humid chamber over night at 4.degree. C. with
primary monoclonal antibody mouse anti-ED1(cD68) (AbD serotec).
After washing with PSB slides were incubated with secondary
antibody donkey anti-mouse (Cy3) for 90 minutes in a dark humid
chamber at room temperature followed by mounting with vectashield.
Samples were analysed using a Zeiss Axioplan-2 imaging microscope
with the digital image-processing program AxioVision 4.8. All
evaluations were done by a single blinded investigator. Analysis of
infiltrating macrophages was done by enumerating ED1 positive cells
in heart (ventricle) and kidney tissue (outer medulla and cortex)
sections. Data were expressed as sum of 20 randomly chosen view
fields, non-overlapping fields per section.
[0518] Statistical analyses were performed with software R for
immunohistochemistry. As the data are nonparametric (counts)
Kruskall-Wallis-tests were applied for overall group differences
while pairwise U-tests with FDR correction were used to analyze
differences between individual groups. P-values below 0.05 were
considered significant.
Results
[0519] FIG. 4 and FIG. 5 show the potential of a robust analog of
17,18-EEQ (Comp-02) to modulate inflammation in dTGR animals. This
was measured both in heart (FIG. 4) and kidney (FIG. 5) with
macrophages infiltration via ED1 staining as a marker for
inflammation. The values shown are expressed as median with
interquartile range. Count values are pooled in bins of 20 view
fields. The results show that dTGR animals have significant higher
amounts of infiltrated macrophages in the cardiac or renal tissue
compared to non-treated SD animals (FIG. 4 and FIG. 5). However,
Comp-02 treatment led to a significant reduction of macrophage
infiltration (ED1 positive cells) in dTGR animals compared to
vehicle treated dTGRs.
* * * * *