U.S. patent application number 13/139260 was filed with the patent office on 2011-12-08 for antagonists of lysophosphatidic acid receptors.
Invention is credited to Jeannie M. Arruda, John Howard Hutchinson, Timothy Parr, Jeffrey Roger Roppe, Thomas Jon Seiders, Bowei Wang.
Application Number | 20110301211 13/139260 |
Document ID | / |
Family ID | 41666879 |
Filed Date | 2011-12-08 |
United States Patent
Application |
20110301211 |
Kind Code |
A1 |
Hutchinson; John Howard ; et
al. |
December 8, 2011 |
ANTAGONISTS OF LYSOPHOSPHATIDIC ACID RECEPTORS
Abstract
Described herein are compounds that are antagonists of
lysophosphatidic receptor(s). Also described are pharmaceutical
compositions and medicaments that include the compounds described
herein, as well as methods of using such antagonists, alone and in
combination with other compounds, for treating LPA-dependent or
LPA-mediated conditions or diseases.
Inventors: |
Hutchinson; John Howard;
(San Diego, CA) ; Seiders; Thomas Jon; (San Diego,
CA) ; Wang; Bowei; (Westfield, NJ) ; Arruda;
Jeannie M.; (San Diego, CA) ; Roppe; Jeffrey
Roger; (Temecula, CA) ; Parr; Timothy; (La
Mesa, CA) |
Family ID: |
41666879 |
Appl. No.: |
13/139260 |
Filed: |
December 15, 2009 |
PCT Filed: |
December 15, 2009 |
PCT NO: |
PCT/US2009/068106 |
371 Date: |
August 29, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61122568 |
Dec 15, 2008 |
|
|
|
Current U.S.
Class: |
514/380 ;
548/245 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 13/02 20180101; A61P 1/04 20180101; A61P 35/02 20180101; A61P
17/02 20180101; A61P 1/18 20180101; A61P 27/02 20180101; A61P 11/08
20180101; A61P 13/00 20180101; A61P 25/04 20180101; A61P 9/00
20180101; A61P 7/10 20180101; A61P 11/06 20180101; A61P 17/06
20180101; A61P 19/04 20180101; C07D 413/10 20130101; A61P 1/00
20180101; A61P 1/16 20180101; A61P 11/00 20180101; A61P 13/12
20180101; A61P 7/12 20180101; A61P 29/00 20180101; A61P 25/02
20180101; A61P 9/10 20180101; A61P 17/00 20180101; A61P 35/00
20180101; C07D 261/14 20130101; A61P 7/04 20180101; A61P 35/04
20180101 |
Class at
Publication: |
514/380 ;
548/245 |
International
Class: |
A61K 31/42 20060101
A61K031/42; C07D 413/10 20060101 C07D413/10; A61K 31/422 20060101
A61K031/422; A61P 1/16 20060101 A61P001/16; A61P 17/00 20060101
A61P017/00; A61P 35/00 20060101 A61P035/00; A61P 9/00 20060101
A61P009/00; A61P 11/00 20060101 A61P011/00; A61P 29/00 20060101
A61P029/00; A61P 1/00 20060101 A61P001/00; A61P 13/12 20060101
A61P013/12; A61P 13/02 20060101 A61P013/02; A61P 13/00 20060101
A61P013/00; A61P 1/18 20060101 A61P001/18; A61P 9/10 20060101
A61P009/10; A61P 7/04 20060101 A61P007/04; A61P 11/06 20060101
A61P011/06; A61P 35/02 20060101 A61P035/02; A61P 35/04 20060101
A61P035/04; A61P 27/02 20060101 A61P027/02; C07D 261/14 20060101
C07D261/14 |
Claims
1. A compound having the structure of Formula (I) or a
pharmaceutically acceptable salt thereof: ##STR00013## wherein
R.sup.1 is --CO.sub.2H, --CO.sub.2R.sup.D, --CN, tetrazolyl,
--C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.10,
--C(.dbd.O)NHSO.sub.2R.sup.10 or
--C(.dbd.O)NHCH.sub.2CH.sub.2SO.sub.3H; R.sup.D is H or
C.sub.1-C.sub.4alkyl; L.sup.1 is absent or C.sub.1-C.sub.6alkylene;
R.sup.3 is H, C.sub.1-C.sub.4alkyl, C.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4-fluoroalkyl; R.sup.7 is H or C.sub.1-C.sub.4alkyl;
R.sup.8 is H, C.sub.1-C.sub.4alkyl, or C.sub.1-C.sub.4-fluoroalkyl;
R.sup.10 is a C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6-fluoroalkyl,
C.sub.3-C.sub.6cycloalkyl, or a substituted or unsubstituted
phenyl; each of R.sup.A, R.sup.B, and R.sup.C are independently
selected from H, F, Cl, Br, I, --CN, --OH, C.sub.1-C.sub.4alkyl,
C.sub.1-C.sub.4-fluoroalkyl, C.sub.1-C.sub.4-fluoroalkoxy,
C.sub.1-C.sub.4alkoxy, and C.sub.1-C.sub.4heteroalkyl; m is 0, 1,
or 2; n is 0, 1, or 2; p is 0, 1, or 2.
2. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein: R.sup.1 is --CO.sub.2H, --CO.sub.2R.sup.D or
--C(.dbd.O)NHSO.sub.2R.sup.10; R.sup.3 is C.sub.1-C.sub.4alkyl;
R.sup.7 is H; R.sup.10 is a C.sub.1-C.sub.6alkyl or a substituted
or unsubstituted phenyl; each R.sup.A is independently selected
from H, F, Cl, Br, I, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3; each R.sup.B is independently selected from H, F,
Cl, Br, I, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3, and
--OCH.sub.3; each R.sup.C is independently selected from H, F, Cl,
Br, I, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3, and --OCH.sub.3;
m is 0 or 1; p is 0 or 1.
3. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein: R.sup.1 is --CO.sub.2H or --CO.sub.2R.sup.D;
R.sup.3 is --CH.sub.3 or --CH.sub.2CH.sub.3; R.sup.8 is H,
--CH.sub.3 or --CF.sub.3; R.sup.D is H, --CH.sub.3, or
--CH.sub.2CH.sub.3.
4. The compound of claim 3, or a pharmaceutically acceptable salt
thereof, wherein: R.sup.1 is --CO.sub.2H; R.sup.3 is --CH.sub.3;
R.sup.8 is --CH.sub.3; L.sup.1 is absent, --CH.sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2CH.sub.2CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH(CH.sub.2CH.sub.3)--,
--C(CH.sub.2CH.sub.3).sub.2--, --CH.sub.2CH(CH.sub.3)--, or
--CH.sub.2C(CH.sub.3).sub.2--.
5. The compound of claim 4, or a pharmaceutically acceptable salt
thereof, wherein: ##STR00014##
6. The compound of claim 5, or a pharmaceutically acceptable salt
thereof, wherein: L.sup.1 is absent, --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH(CH.sub.2CH.sub.3)--,
or --C(CH.sub.2CH.sub.3).sub.2--; each R.sup.C is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3; m is 0; n is 0, 1, or 2; p is 0.
7. The compound of claim 6, or a pharmaceutically acceptable salt
thereof, wherein: L.sup.1 is absent or --CH.sub.2--; each R.sup.C
is independently selected from H, F, Cl, --CH.sub.3, --CF.sub.3,
and --OH; n is 0 or 1.
8. The compound of claim 4, or a pharmaceutically acceptable salt
thereof, wherein: ##STR00015##
9. The compound of claim 8, or a pharmaceutically acceptable salt
thereof, wherein: L.sup.1 is absent, --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH(CH.sub.2CH.sub.3)--,
or --C(CH.sub.2CH.sub.3).sub.2--; each R.sup.C is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3; m is 0; n is 0, 1, or 2; p is 0.
10. The compound of claim 9, or a pharmaceutically acceptable salt
thereof, wherein: L.sup.1 is absent or --CH.sub.2--; each R.sup.C
is independently selected from H, F, Cl, --CH.sub.3, --CF.sub.3,
and --OH; n is 0 or 1.
11. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein: R.sup.1 is --C(.dbd.O)NHSO.sub.2R.sup.10 R.sup.3
is --CH.sub.3 or --CH.sub.2CH.sub.3; R.sup.8 is H, --CH.sub.3 or
--CF.sub.3; R.sup.10 is --CH.sub.3, or --CH.sub.2CH.sub.3.
12. The compound of claim 1 selected from:
(4'-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 1);
(4'-{3-Methyl-4-[1-(3-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 2);
(4'-{4-[1-(2,4-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 3);
(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic acid (Compound 4);
(4'-{4-[1-(3-Bromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-4-yl)-acetic acid (Compound 5);
(4'-{4-[1-(2-Methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-
-biphenyl-4-yl)-acetic acid (Compound 6);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
6-methoxy-biphenyl-3-yl)-acetic acid (Compound 7);
4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-4-carboxylic acid (Compound 8);
4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-2-carboxylic acid (Compound 9);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-2-yl)-acetic acid (Compound 10);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic acid (Compound 11);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-3-yl)-acetic acid (Compound 12);
3-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid (Compound 13);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
6-fluoro-biphenyl-3-yl)-acetic acid (Compound 14);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
4-fluoro-biphenyl-3-yl)-acetic acid (Compound 15);
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic acid methyl ester (Compound 16);
2-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid ethyl ester (Compound 17);
2-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid (Compound 18);
2-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 19);
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid (Compound 20);
4-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-butyric acid (Compound 21);
4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-3-carboxylic acid (Compound 22);
(4'-{4-[1-(4-Chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 23);
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 24);
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-2'-methyl-biphenyl-4-yl)-acetic acid (Compound 25);
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 26);
(4'-{4-[(R)-1-(2-Chloro-phenye-ethoxycarbonylamino]-3-methyl-isoxazol-5-y-
l}-biphenyl-4-yl)-acetic acid (Compound 27);
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 28);
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 29);
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic acid (Compound 30);
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic acid (Compound 31);
(4'-{4-[1-(2,6-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 32);
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-2'-methyl-biphenyl-4-yl)-propionic acid (Compound 33);
(4'-{4-[(S)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 34);
(4'-{4-[(S)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 35);
{4'-[4-(2-Chloro-benzyloxycarbonylamino)-3-methyl-isoxazol-5-yl]-biphenyl-
-4-yl}-acetic acid (Compound 36);
{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-4-yl}-acetic acid (Compound 37);
(4'-{4-[1-(2,3-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 38);
(4'-{4-[1-(2,4-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 39);
(4'-{4-[1-(2-Fluoro-4-methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxa-
zol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 40);
(4'-{4-[1-(2,5-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 41);
(4'-{4-[1-(2,6-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 42);
{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-3-yl}-acetic acid (Compound 43);
4'-[3-Methyl-44(R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl--
4-carboxylic acid (Compound 44);
{4'-[3-Methyl-4-(R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-
-2-yl}-acetic acid (Compound 45);
{4'-[3-Methyl-4-((R)-1-o-tolyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphen-
yl-4-yl}-acetic acid (Compound 46);
2-(4'-{4-[(R,R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazo-
l-5-yl}-biphenyl-4-yl)-propionic acid (Compound 47);
2-(4'-{4-[(R,S)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazo-
l-5-yl}-biphenyl-4-yl)-propionic acid (Compound 48);
(3'-Chloro-4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-i-
soxazol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 49);
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-butyric acid (Compound 50);
(2'-Chloro-4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-i-
soxazol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 51);
(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-2'-fluoro-biphenyl-4-yl)-acetic acid (Compound 52);
4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-y-
l}-biphenyl-4-carboxylic acid (Compound 53);
(4'-{4-[(R)-1-(3,5-Dibromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-
-5-yl}-biphenyl-4-yl)-acetic acid (Compound 56);
{4'-[3-Methyl-4-((S)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl}-bipheny-
l-4-yl]-acetic acid (Compound 57);
(4'-{4-[(R)-1-(3-Hydroxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-
-yl}-biphenyl-4-yl)-acetic acid (Compound 58);
{4'-[3-Methyl-4-(1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4--
yl}-acetic acid (Compound 59);
[5-(4'-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-chloro-phenyl)-ethyl ester (Compound 61);
[5-(4'-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 62);
{3-Methyl-5-[4'-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl}-isoxazol-4-yl]-ca-
rbamic acid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 63);
{3-Methyl-5-[4'-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-ca-
rbamic acid (R)-1-(2-chloro-phenyl)-ethyl ester (Compound 64);
[5-(4'-Carbamimidoylmethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbami-
c acid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 65);
{5-[4'-(2-Acetylamino-2-imino-ethyl)-biphenyl-4-yl]-3-methyl-isoxazol-4-y-
l}-carbamic acid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 66);
and
2-(2-{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bi-
phenyl-4-yl}-acetylamino)-ethanesulfonic acid (Compound 67); or a
pharmaceutically acceptable salt thereof.
13. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1, or a pharmaceutically
acceptable salt thereof, and at least one pharmaceutically
acceptable inactive ingredient.
14. The pharmaceutical composition of claim 13, wherein the
pharmaceutical composition is: (a) formulated for intravenous
injection, subcutaneous injection, oral administration, inhalation,
nasal administration, topical administration, ophthalmic
administration or otic administration; or (b) a tablet, a pill, a
capsule, a liquid, an inhalant, a nasal spray solution, a
suppository, a suspension, a gel, a colloid, a dispersion, a
suspension, a solution, an emulsion, an ointment, a lotion, an eye
drop or an ear drop.
15. A method: (a) of inhibiting the physiological activity of LPA
in a mammal; (b) for treating or preventing a LPA-dependent or
LPA-mediated disease or condition in a mammal; (c) for treating or
preventing fibrosis of organs or tissues, scarring, liver diseases,
dermatological conditions, cancer, cardiovascular diseases,
respiratory diseases or conditions, inflammatory diseases,
gastrointestinal tract diseases, renal diseases, urinary
tract-associated diseases, inflammatory diseases of lower urinary
tract, dysuria, frequent urination, pancreas disease, arterial
obstruction, cerebral infarction, cerebral hemorrhage, pain,
peripheral neuropathy, or fibromyalgia in a mammal; or (d) for
treating or preventing lung fibrosis, asthma, chronic obstructive
pulmonary disease (COPD), renal fibrosis, acute kidney injury,
chronic kidney disease, liver fibrosis, skin fibrosis, fibrosis of
the gut, breast cancer, pancreatic cancer, ovarian cancer, prostate
cancer, glioblastoma, bone cancer, colon cancer, bowel cancer, head
and neck cancer, melanoma, multiple myeloma, chronic lymphocytic
leukemia, cancer pain, tumor metastasis, transplant organ
rejection, scleroderma, ocular fibrosis, age related macular
degeneration (AMD), diabetic retinopathy, collagen vascular
disease, atherosclerosis, or neuropathic pain in a mammal;
comprising administering a therapeutically effective amount of a
compound according to claim 1, or a pharmaceutically acceptable
salt thereof to the mammal in need thereof.
Description
RELATED APPLICATIONS
[0001] This application claims benefit of U.S. Provisional
Application No. 61/122,568, entitled "ANTAGONISTS OF
LYSOPHOSPHATIDIC ACID RECEPTORS" filed on Dec. 15, 2008, which is
herein incorporated by reference.
FIELD OF THE INVENTION
[0002] Described herein are compounds, methods of making such
compounds, pharmaceutical compositions and medicaments comprising
such compounds, and methods of using such compounds to treat,
prevent or diagnose diseases, disorders or conditions associated
with one or more of the lysophosphatidic acid (LPA) receptors.
BACKGROUND OF THE INVENTION
[0003] Lysophospholipids are membrane-derived bioactive lipid
mediators. Lysophospholipids affect fundamental cellular functions
that include proliferation, differentiation, survival, migration,
adhesion, invasion, and morphogensis. These functions influence
many biological processes that include, but are not limited to,
neurogensis, angiogenesis, wound healing, fibrosis, immunity, and
carcinogenesis.
[0004] Lysophosphatidic acid (LPA) is a lysophospholipid that has
been shown to act through sets of specific G protein-coupled
receptors (GPCRs) in an autocrine and paracrine fashion. LPA
binding to its cognate GPCRs (LPA.sub.1, LPA.sub.2, LPA.sub.3,
LPA.sub.4, LPA.sub.5, LPA.sub.6) activates intracellular signaling
pathways to produce a variety of biological responses. Antagonists
of the LPA receptors find use in the treatment of diseases,
disorders or conditions in which LPA plays a role.
SUMMARY OF THE INVENTION
[0005] In one aspect, presented herein are compounds of Formula (I)
that inhibit the physiological activity of lysophosphatidic acid
(LPA), and therefore, are useful as agents for the treatment or
prevention of diseases in which inhibition of the physiological
activity of LPA is useful, such as diseases in which an LPA
receptor participates, is involved in the etiology or pathology of
the disease, or is otherwise associated with at least one symptom
of the disease. In a related aspect, such compounds are useful as
agents for the treatment or prevention of side effects,
complications, or adverse events associated with the use of a
different therapeutic agent or therapeutic action (e.g., radiation,
surgery, etc) used in treating a disease or condition.
[0006] In one aspect, the compounds of Formula (I) are useful for
the treatment of fibrosis of organs (liver, kidney, lung, heart and
the like), liver diseases (acute hepatatis, chronic hepatitis,
liver fibrosis, liver cirrhosis, portal hypertension, regenerative
failure, non-alcoholic steatohepatitis (NASH), liver hypofunction,
hepatic blood flow disorder, and the like), cell proliferative
disease (cancer (solid tumor, solid tumor metastasis, vascular
fibroma, myeloma, multiple myeloma, Kaposi's sarcoma, leukemia,
chronic lymphocytic leukemia (CLL) and the like) and invasive
metastasis of cancer cell, and the like), inflammatory disease
(psoriasis, nephropathy, pneumonia and the like), gastrointestinal
tract disease (irritable bowel syndrome (IBS), inflammatory bowel
disease (IBD), abnormal pancreatic secretion, and the like), renal
disease, urinary tract-associated disease (benign prostatic
hyperplasia or symptoms associated with neuropathic bladder
disease, spinal cord tumor, hernia of intervertebral disk, spinal
canal stenosis, symptoms derived from diabetes, lower urinary tract
disease (obstruction of lower urinary tract, and the like),
inflammatory disease of lower urinary tract, dysuria, frequent
urination, and the like), pancreas disease, abnormal
angiogenesis-associated disease (arterial obstruction and the
like), scleroderma, brain-associated disease (cerebral infarction,
cerebral hemorrhage, and the like), neuropathic pain, peripheral
neuropathy, and the like, ocular disease (age-related macular
degeneration (AMD), diabetic retinopathy, proliferative
vitreoretinopathy (PVR), cicatricial pemphigoid, glaucoma
filtration surgery scarring, and the like). In one aspect, the
compounds of Formula (I) are used in the treatment of fibrotic
diseases or conditions.
[0007] In one aspect, described herein are compounds of Formula
(I), pharmaceutically acceptable salts, solvates, and prodrugs
thereof. Compounds of Formula (I) are antagonists of at least one
of the LPA receptors selected from LPA.sub.1, LPA.sub.2, LPA.sub.3,
LPA.sub.4, LPA.sub.5 and LPA.sub.6. In one embodiment, compounds of
Formula (I) are antagonists of LPA.sub.1. In one embodiment,
compounds of Formula (I) are antagonists of LPA.sub.1 and/or
LPA.sub.3. In some embodiments, compounds of Formula (I) are
antagonists of LPA.sub.1 and/or LPA.sub.2. In some embodiments,
compounds of Formula (I) are selective antagonists for one of the
LPA receptors relative to the other LPA receptors. In some
embodiments, such a selective antagonist is selective for the
LPA.sub.1 receptor. In some embodiments, such a selective
antagonist is selective for the LPA.sub.2 receptor. In some
embodiments, such a selective antagonist is selective for the
LPA.sub.3 receptor.
[0008] Compounds of Formula (I) are used in the treatment of
diseases, disorders, or conditions in which activation of at least
one LPA receptor by LPA contributes to the symptomology or
progression of the disease, disorder or condition. In one aspect,
the methods, compounds, pharmaceutical compositions, and
medicaments described herein comprise antagonists of LPA receptors.
In one aspect, the methods, compounds, pharmaceutical compositions,
and medicaments described herein comprise antagonists of LPA.sub.1,
LPA.sub.2, or LPA.sub.3, or combinations thereof.
[0009] In one aspect, provided herein is a compound of Formula (I),
or a pharmaceutically acceptable salt thereof:
##STR00001##
[0010] wherein [0011] R.sup.1 is --CO.sub.2H, --CO.sub.2R.sup.D,
--CN, tetrazolyl, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.10,
--C(.dbd.O)NHSO.sub.2R.sup.10 or
--C(.dbd.O)NHCH.sub.2CH.sub.2SO.sub.3H; R.sup.D is H or
C.sub.1-C.sub.4alkyl; [0012] L.sup.1 is absent or
C.sub.1-C.sub.6alkylene; [0013] R.sup.3 is H, C.sub.1-C.sub.4alkyl,
C.sub.3-C.sub.6cycloalkyl, or C.sub.1-C.sub.4-fluoroalkyl; [0014]
R.sup.7 is H or C.sub.1-C.sub.4alkyl; [0015] R.sup.8 is H,
C.sub.1-C.sub.4alkyl, or C.sub.1-C.sub.4-fluoroalkyl; [0016]
R.sup.10 is a C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6-fluoroalkyl,
C.sub.3-C.sub.6cycloalkyl, or a substituted or unsubstituted
phenyl; [0017] each of R.sup.A, R.sup.B, and R.sup.C are
independently selected from H, F, Cl, Br, I, --CN, --OH,
C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.4-fluoroalkyl,
C.sub.1-C.sub.4-fluoroalkoxy, C.sub.1-C.sub.4alkoxy, and
C.sub.1-C.sub.4heteroalkyl; [0018] m is 0, 1, or 2; n is 0, 1, or
2; p is 0, 1, or 2.
[0019] In one aspect, provided is are compounds presented in Table
1, Table 2, Table 3, FIG. 1, FIG. 2, FIG. 3, FIG. 4, FIG. 5, FIG.
6, and FIG. 7.
[0020] Compounds of Formula (I) are antagonists of at least one LPA
receptor. In some embodiments, the compound of Formula (I) is an
antagonist of LPA.sub.1. In some embodiments, the compound of
Formula (I) is an antagonist of LPA.sub.2. In some embodiments, the
compound of Formula (I) is an antagonist of LPA.sub.3.
[0021] In some embodiments, presented herein are compounds selected
from active metabolites, tautomers, solvates, pharmaceutically
acceptable salts or prodrugs of a compound of Formula (I).
[0022] In some embodiments, provided is a pharmaceutical
composition comprising a therapeutically effective amount of a
compound of Formula (I). In some embodiments, the pharmaceutical
composition also contains at least one pharmaceutically acceptable
inactive ingredient.
[0023] In some embodiments, provided is a pharmaceutical
composition comprising a therapeutically effective amount of a
compound of Formula (I), or a pharmaceutically acceptable salt
thereof, and at least one pharmaceutically acceptable inactive
ingredient. In one aspect, the pharmaceutical composition is
formulated for intravenous injection, subcutaneous injection, oral
administration, inhalation, nasal administration, topical
administration, ophthalmic administration or otic administration.
In some embodiments, the pharmaceutical composition is a tablet, a
pill, a capsule, a liquid, an inhalant, a nasal spray solution, a
suppository, a suspension, a gel, a colloid, a dispersion, a
suspension, a solution, an emulsion, an ointment, a lotion, an eye
drop or an ear drop.
[0024] In some embodiments, the pharmaceutical composition further
comprises one or more additional therapeutically active agents
selected from: corticosteroids, immunosuppresants, analgesics,
anti-cancer agent, anti-inflammatories, chemokine receptor
antagonists, bronchodilators, leukotriene receptor antagonists,
leukotriene formation inhibitors, monoacylglycerol kinase
inhibitors, phospholipase A.sub.1 inhibitors, phospholipase A.sub.2
inhibitors, and lysophospholipase D (lysoPLD) inhibitors, autotaxin
inhibitors, decongestants, antihistamines, mucolytics,
anticholinergics, antitussives, expectorants, and .beta.-2
agonists.
[0025] In some embodiments, provided is a method comprising
administering a compound of Formula (I) to a human with a
LPA-dependent or LPA-mediated disease or condition. In some
embodiments, the human is already being administered one or more
additional therapeutically active agents other than a compound of
Formula (I). In some embodiments, the method further comprises
administering one or more additional therapeutically active agents
other than a compound of Formula (I).
[0026] In some embodiments, the one or more additional
therapeutically active agents other than a compound of Formula (I)
are selected from: corticosteroids, immunosuppresants, analgesics,
anti-cancer agent, anti-inflammatories, chemokine receptor
antagonists, bronchodilators, leukotriene receptor antagonists,
leukotriene formation inhibitors, monoacylglycerol kinase
inhibitors, phospholipase A.sub.1 inhibitors, phospholipase A.sub.2
inhibitors, and lysophospholipase D (lysoPLD) inhibitors, autotaxin
inhibitors, decongestants, antihistamines, mucolytics,
anticholinergics, antitussives, expectorants, and .beta.-2
agonists.
[0027] In another aspect is the use of a compound of Formula (I) in
the manufacture of a medicament for treating a disease, disorder or
condition in which the activity of at least one LPA receptor
contributes to the pathology and/or symptoms of the disease or
condition. In one embodiment of this aspect, the LPA receptor is
selected from LPA.sub.1, LPA.sub.2, LPA.sub.3, LPA.sub.4, LPA.sub.5
and LPA.sub.6. In some embodiments, the LPA receptor is LPA.sub.1.
In some embodiments, the LPA receptor is LPA.sub.2. In some
embodiments, the LPA receptor is LPA.sub.3. In some embodiments,
the disease or condition is any of the diseases or conditions
specified herein.
[0028] Also provided is a method of inhibiting the physiological
activity of LPA in a mammal comprising administering a
therapuetically effective amount of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof to the mammal in need
thereof.
[0029] In one aspect, provided is a medicament for treating a
LPA-dependent or LPA-mediated disease or condition in a mammal
comprising a therapeutically effective amount of a compound of
Formula (I).
[0030] In some cases disclosed herein is the use of a compound of
Formula (I) in the manufacture of a medicament for the treatment of
a LPA-dependent or LPA-mediated disease or condition.
[0031] In some cases disclosed herein is the use of a compound of
Formula (I) in the treatment or prevention of a LPA-dependent or
LPA-mediated disease or condition.
[0032] In one aspect, is a method for treating or preventing a
LPA-dependent or LPA-mediated disease or condition in a mammal
comprising administering a therapuetically effective amount of a
compound of Formula (I).
[0033] In one aspect, LPA-dependent or LPA-mediated diseases or
conditions include, but are not limited to, fibrosis of organs or
tissues, scarring, liver diseases, dermatological conditions,
cancer, cardiovascular disease, respiratory diseases or conditions,
inflammatory disease, gastrointestinal tract disease, renal
disease, urinary tract-associated disease, inflammatory disease of
lower urinary tract, dysuria, frequent urination, pancreas disease,
arterial obstruction, cerebral infarction, cerebral hemorrhage,
pain, peripheral neuropathy, and fibromyalgia.
[0034] In some embodiments, the LPA-dependent or LPA-mediated
disease or condition is selected from idiopathic pulmonary
fibrosis; other diffuse parenchymal lung diseases of different
etiologies including iatrogenic drug-induced fibrosis, occupational
and/or environmental induced fibrosis, granulomatous diseases
(sarcoidosis, hypersensitivity pneumonia), collagen vascular
disease, alveolar proteinosis, langerhans cell granulomatosis,
lymphangioleiomyomatosis, inherited diseases (Hermansky-Pudlak
Syndrome, tuberous sclerosis, neurofibromatosis, metabolic storage
disorders, familial interstitial lung disease); radiation induced
fibrosis; chronic obstructive pulmonary disease (COPD);
scleroderma; bleomycin induced pulmonary fibrosis; chronic asthma;
silicosis; asbestos induced pulmonary fibrosis; acute respiratory
distress syndrome (ARDS); kidney fibrosis; tubulointerstitium
fibrosis; glomerular nephritis; focal segmental glomerular
sclerosis; IgA nephropathy; hypertension; Alport; gut fibrosis;
liver fibrosis; cirrhosis; alcohol induced liver fibrosis;
toxic/drug induced liver fibrosis; hemochromatosis; nonalcoholic
steatohepatitis (NASH); biliary duct injury; primary biliary
cirrhosis; infection induced liver fibrosis; viral induced liver
fibrosis; and autoimmune hepatitis; corneal scarring; hypertrophic
scarring; Duputren disease, keloids, cutaneous fibrosis; cutaneous
scleroderma; spinal cord injury/fibrosis; myelofibrosis; vascular
restenosis; atherosclerosis; arteriosclerosis; Wegener's
granulomatosis; Peyronie's disease, chronic lymphocytic leukemia,
tumor metastasis, transplant organ rejection, endometreosis,
neonatal respiratory distress syndrome and neuropathic pain.
[0035] In one aspect, provided is a method for the treatment or
prevention of organ fibrosis in a mammal comprising administering a
therapeutically effective amount of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof to a mammal in need
thereof.
[0036] In some embodiments, the organ fibrosis comprises lung
fibrosis, renal fibrosis, or hepatic fibrosis.
[0037] In one aspect, provided is a method of improving lung
function in a mammal comprising administering a therapeutically
effective amount of a compound of Formula (I) or a pharmaceutically
acceptable salt thereof to the mammal in need thereof. In one
aspect, the mammal has been diagnosed as having lung fibrosis.
[0038] In one aspect, compounds disclosed herein are used to treat
idiopathic pulmonary fibrosis (usual interstitial pneumonia) in a
mammal.
[0039] In some embodiments, compounds disclosed herein are used to
treat diffuse parenchymal interstitial lung diseases in mammal:
iatrogenic drug induced, occupational/environmental (Farmer lung),
granulomatous diseases (sarcoidosis, hypersensitivity pneumonia),
collagen vascular disease (scleroderma and others), alveolar
proteinosis, langerhans cell granulonmatosis,
lymphangioleiomyomatosis, Hermansky-Pudlak Syndrome, Tuberous
sclerosis, neurofibromatosis, metabolic storage disorders, familial
interstitial lung disease.
[0040] In some embodiments, compounds disclosed herein are used to
treat post-transplant fibrosis associated with chronic rejection in
a mammal: Bronchiolitis obliterans for lung transplant.
[0041] In some embodiments, compounds disclosed herein are used to
treat cutaneous fibrosis in a mammal: cutaneous scleroderma,
Dupuytren disease, keloids.
[0042] In one aspect, compounds disclosed herein are used to treat
hepatic fibrosis with or without cirrhosis in a mammal: toxic/drug
induced (hemochromatosis), alcoholic liver disease, viral hepatitis
(hepatitis B virus, hepatitis C virus, HCV), nonalcoholic liver
disease (NASH), metabolic and auto-immune.
[0043] In one aspect, compounds disclosed herein are used to treat
renal fibrosis in a mammal: tubulointerstitium fibrosis, glomerular
sclerosis.
[0044] In any of the aforementioned aspects involving the treatment
of LPA dependent diseases or conditions are further embodiments
comprising administering at least one additional agent in addition
to the administartion of a compound having the structure of Formula
(I). In various embodiments, each agent is administered in any
order, including simultaneously.
[0045] In any of the embodiments disclosed herein, the mammal is a
human.
[0046] In some embodiments, compounds provided herein are
administered to a human. In some embodiments, compounds provided
herein are orally administered to a human.
[0047] In some embodiments, compounds provided herein are used as
antagonists of at least one LPA receptor. In some embodiments,
compounds provided herein are used for inhibiting the activity of
at least one LPA receptor or for the treatment of a disease or
condition that would benefit from inhibition of the activity of at
least one LPA receptor. In one aspect, the LPA receptor is
LPA.sub.1.
[0048] In other embodiments, compounds provided herein are used for
the formulation of a medicament for the inhibition of LPA.sub.1
activity.
[0049] Other objects, features and advantages of the compounds,
methods and compositions described herein will become apparent from
the following detailed description. It should be understood,
however, that the detailed description and the specific examples,
while indicating specific embodiments, are given by way of
illustration only, since various changes and modifications within
the spirit and scope of the instant disclosure will become apparent
to those skilled in the art from this detailed description
BRIEF DESCRIPTION OF THE FIGURES
[0050] FIG. 1. Illustrative examples of compounds described
herein.
[0051] FIG. 2. Illustrative examples of compounds described
herein.
[0052] FIG. 3. Illustrative examples of compounds described
herein.
[0053] FIG. 4. Illustrative examples of compounds described
herein.
[0054] FIG. 5. Illustrative examples of compounds described
herein.
[0055] FIG. 6. Illustrative examples of compounds described
herein.
[0056] FIG. 7. Illustrative examples of compounds described
herein.
DETAILED DESCRIPTION OF THE INVENTION
[0057] Lysophospholipids (such as lysophosphatidic acid (LPA))
affect fundamental cellular functions that include cellular
proliferation, differentiation, survival, migration, adhesion,
invasion, and morphogensis. These functions influence many
biological processes that include neurogensis, angiogenesis, wound
healing, immunity, and carcinogenesis.
[0058] LPA acts through sets of specific G protein-coupled
receptors (GPCRs) in an autocrine and paracrine fashion. LPA
binding to its cognate GPCRs (LPA.sub.1, LPA.sub.2, LPA.sub.3,
LPA.sub.4, LPA.sub.5, LPA.sub.6) activates intracellular signaling
pathways to produce a variety of biological responses.
[0059] LPA has a role as a biological effector molecule, and has a
diverse range of physiological actions such as, but not limited to,
effects on blood pressure, platelet activation, and smooth muscle
contraction, and a variety of cellular effects, which include cell
growth, cell rounding, neurite retraction, and actin stress fiber
formation and cell migration. The effects of LPA are predominantly
receptor mediated.
[0060] Activation of the LPA receptors (LPA.sub.1, LPA.sub.2,
LPA.sub.3, LPA.sub.4, LPA.sub.5, LPA.sub.6) with LPA mediates a
range of downstream signaling cascades. The actual pathway and
realized end point are dependent on a range of variables that
include receptor usage, cell type, expression level of a receptor
or signaling protein, and LPA concentration. Nearly all mammalian
cells, tissues and organs co-express several LPA-receptor subtypes,
which indicates that LPA receptors signal in a cooperative manner.
LPA.sub.1, LPA.sub.2, and LPA.sub.3 share high amino acid sequence
similarity.
Illustrative Biological Activity
[0061] LPA regulates many important functions of fibroblasts in
wound healing, including proliferation, migration, differentiation
and contraction. Fibroblast proliferation is required in wound
healing in order to fill an open wound. In contrast, fibrosis is
characterized by intense proliferation and accumulation of
myofibroblasts that actively synthesize ECM and proinflammatory
cytokines LPA can either increase or suppress the proliferation of
cell types important in wound healing.
[0062] Tissue injury initiates a complex series of host
wound-healing responses; if successful, these responses restore
normal tissue structure and function. If not, these responses can
lead to tissue fibrosis and loss of function.
[0063] A number of muscular dystrophies are characterized by a
progressive weakness and wasting of musculature, and by extensive
fibrosis. It has been shown that LPA treatment of cultured
myoblasts induced significant expression of connective tissue
growth factor (CTGF). CTGF subsequently induces collagen,
fibronectin and integrin expression and induces dedifferentiation
of these myoblasts. Treatment of a variety of cell types with LPA
induces reproducible and high level induction of CTGF. CTGF is a
profibrotic cytokine, signaling down-stream and in parallel with
TGF.beta..
[0064] LPA and LPA.sub.1 play key pathogenic roles in pulmonary
fibrosis. Fibroblast chemoattractant activity plays an important
role in the lungs in patients with pulmonary fibrosis. Profibrotic
effects of LPA.sub.1-receptor stimulation is explained by
LPA.sub.1-receptor-mediated vascular leakage and increased
fibroblast recruitment, both profibrotic events. The LPA-LPA.sub.1
pathway has a role in mediating fibroblast migration and vascular
leakage in IPF. The end result is the aberrant healing process that
characterises this fibrotic condition.
[0065] The LPA-LPA2 pathway contributes to the activation of the
TGF-.beta. pathway in pulmonary fibrosis. In some embodiments,
compounds that inhibit LPA2 show efficacy in the treatment of lung
fibrosis. In some embodiments, compounds that inhibit both LPA1 and
LPA2 show improved efficacy in the treatment of lung fibrosis
compared to compounds which inhibit only LPA1 or LPA2.
[0066] LPA and LPA.sub.1 are involved in the etiology of kidney
fibrosis. In mice invalidated for the LPA.sub.1 receptor (LPA.sub.1
(-/-), the development of renal fibrosis was significantly
attenuated. Unilateral ureteral obstruction (UUO; animal model of
renal fibrosis) mice treated with the LPA receptor antagonist
Ki16425 closely resembled the LPA.sub.1 (-/-) mice.
[0067] LPA is implicated in liver disease and fibrosis. Plasma LPA
levels and serum autotoxin are elevated in hepatitis patients and
animal models of liver injury in correlation with increased
fibrosis. LPA also regulates liver cell function. LPA.sub.1 and
LPA.sub.2 receptors are expressed by mouse hepatic stellate cells
and LPA stimulates migration of hepatic myofibroblasts.
[0068] LPA is in involved in wound healing in the eye. LPA.sub.1
and LPA.sub.3 receptors are detectable in the normal rabbit corneal
epithelial cells, keratocytes and endothelial cells and LPA.sub.1
and LPA.sub.3 expression are increased in corneal epithelial cells
following injury.
[0069] LPA is present in the aqueous humor and the lacrimal gland
fluid of the rabbit eye and these levels are increased in a rabbit
corneal injury model.
[0070] LPA induces actin stress fiber formation in rabbit corneal
endothelial and epithelial cells and promotes contraction corneal
fibroblasts. LPA also stimulates proliferation of human retinal
pigmented epithelial cells.
[0071] LPA is implicated in myocardial infarction and cardiac
fibrosis. Serum LPA levels are increased in patients following
mycocardial infarction (MI) and LPA stimulates proliferation and
collagen production (fibrosis) by rat cardiac fibroblasts. Both
LPA1 and LPA3 receptors are highly expressed in human heart
tissue.
[0072] In one aspect, compounds of Formula (I) are used to treat or
prevent fibrosis in a mammal. In one aspect, compounds of Formula
(I) are used to treat or prevent fibrosis of an organ or tissue in
a mammal.
[0073] The terms "fibrosis" or "fibrosing disorder," as used
herein, refers to conditions that are associated with the abnormal
accumulation of cells and/or fibronectin and/or collagen and/or
increased fibroblast recruitment and include but are not limited to
fibrosis of individual organs or tissues such as the heart, kidney,
liver, joints, lung, pleural tissue, peritoneal tissue, skin,
cornea, retina, musculoskeletal and digestive tract.
[0074] Exemplary diseases, disorders, or conditions that involve
fibrosis include, but are not limited to: Lung diseases associated
with fibrosis, e.g., idiopathic pulmonary fibrosis, pulmonary
fibrosis secondary to systemic inflammatory disease such as
rheumatoid arthritis, scleroderma, lupus, cryptogenic fibrosing
alveolitis, radiation induced fibrosis, chronic obstructive
pulmonary disease (COPD), scleroderma, chronic asthma, silicosis,
asbestos induced pulmonary or pleural fibrosis, acute lung injury
and acute respiratory distress (including bacterial pneumonia
induced, trauma induced, viral pneumonia induced, ventilator
induced, non-pulmonary sepsis induced, and aspiration induced);
Chronic nephropathies associated with injury/fibrosis (kidney
fibrosis), e.g., glomerulonephritis secondary to systemic
inflammatory diseases such as lupus and scleroderma, diabetes,
glomerular nephritis, focal segmental glomerular sclerosis, IgA
nephropathy, hypertension, allograft and Alport; Gut fibrosis,
e.g., scleroderma, and radiation induced gut fibrosis; Liver
fibrosis, e.g., cirrhosis, alcohol induced liver fibrosis,
nonalcoholic steatohepatitis (NASH), biliary duct injury, primary
biliary cirrhosis, infection or viral induced liver fibrosis (e.g.,
chronic HCV infection), and autoimmune hepatitis; Head and neck
fibrosis, e.g., radiation induced; Corneal scarring, e.g., LASIK
(laser-assisted in situ keratomileusis), corneal transplant, and
trabeculectomy; Hypertrophic scarring and keloids, e.g., burn
induced or surgical; and Other fibrotic diseases, e.g.,
sarcoidosis, scleroderma, spinal cord injury/fibrosis,
myelofibrosis, vascular restenosis, atherosclerosis,
arteriosclerosis, Wegener's granulomatosis, mixed connective tissue
disease, and Peyronie's disease.
[0075] In one aspect, a mammal suffering from one of the following
non-limiting exemplary diseases, disorders, or conditions will
benefit from therapy with a compound of Formula (I):
atherosclerosis, thrombosis, heart disease, vasculitis, formation
of scar tissue, restenosis, phlobitis, COPD (chronic obstructive
pulmonary disease), pulmonary hypertension, pulmonary fibrosis,
pulmonary inflammation, bowel adhesions, bladder fibrosis and
cystitis, fibrosis of the nasal passages, sinusitis, inflammation
mediated by neutrophils, and fibrosis mediated by fibroblasts.
[0076] In one aspect, compounds of Formula (I) are used to treat a
dermatological disorders in a mammal. Dermatological disorders
include, but are not limited to, proliferative or inflammatory
disorders of the skin such as, atopic dermatitis, bullous
disorders, collagenoses, psoriasis, psoriatic lesions, dermatitis,
contact dermatitis, eczema, urticaria, rosacea, wound healing,
scarring, hypertrophic scarring, keloids, Kawasaki Disease,
rosacea, Sjogren-Larsso Syndrome, urticaria.
[0077] In some embodiments, provided is a method of reducing lung
injury, vascular leakage, inflammation and/or fibrosis in a mammal
comprising administering to the mammal a selective LPA1 receptor
antagonist. In some embodiments, provided is a method of reducing
lung injury, vascular leakage, inflammation and fibrosis in a
mammal comprising administering to the mammal a selective LPA1
receptor antagonist. In some embodiments, provided is a method of
attenuating fibrosis in a mammal comprising administering a
selective LPA1 receptor antagonist. In some embodiments, provided
is a method of attenuating tissue remodeling and fibrosis in a
mammal comprising administering a selective LPA1 receptor
antagonist.
[0078] In some embodiments, provided is a method of decreasing
cytokine production in a mammal comprising administering a
selective LPA1 receptor antagonist. In some embodiments, the method
of decreasing cytokine production in a mammal comprising
administering a selective LPA1 receptor antagonist results in a
reduction of tissue damage and fibrosis in a mammal.
[0079] In some embodiments, provided is a method of treating
fibrosis is a mammal comprising administering to the mammal a
selective LPA1 receptor antagonist. In some embodiments, provided
is a method of treating fibrosis in a mammal while maintaining body
weight in the mammal comprising administering to the mammal a
selective LPA1 receptor antagonist. In some embodiments, provided
is a method of treating respiratory disease in a mammal comprising
administering to the mammal a selective LPA1 receptor
antagonist.
[0080] In some embodiments, provided is a method of treating
fibrosis in a mammal with a selective LPA1 receptor anatgonist,
wherein the fibrosis in the mammal is not responsive to treatment
with pirfenidone. In some embodiments, the LPA1 receptor antagonist
is a compound of Formula (I).
[0081] As shown in the Examples, a selective LPA1 receptor
antagonist reduced lung fibrosis, kidney fibrosis and liver
fibrosis in various animal models of fibrosis.
[0082] LPA is released following tissue injury. LPA.sub.1 plays a
role in the initiation of neuropathic pain. In one aspect,
compounds of Formula (I) are used in the treatment of pain in a
mammal. In one aspect, the pain is acute pain or chronic pain. In
another aspect, the pain is neuropathic pain. In another aspect,
the pain is cancer pain. In one aspect, compounds of Formula (I)
are used in the treatment of fibromylagia.
[0083] Lysophospholipid receptor signaling plays a role in the
etiology of cancer. Lysophosphatidic acid (LPA) and its G
protein-coupled receptors (GPCRs) LPA.sub.1, LPA.sub.2, and/or
LPA.sub.3 play a role in the development of several types of
cancers.
[0084] LPA contributes to tumorigenesis by increasing motility and
invasiveness of cells. LPA has been implicated in the initiation or
progression of ovarian cancer. LPA is present at significant
concentrations (2-80 .mu.M) in the ascitic fluid of ovarian cancer
patients. LPA receptors (LPA2 and LPA3) are also overexpressed in
ovarian cancer cells as compared to normal ovarian surface
epithelial cells. LPA has also been implicated in the initiation or
progression of prostate cancer, breast cancer, melanoma, head and
neck cancer, bowel cancer (colorectal cancer), thyroid cancer,
glioblastoma, and other cancers.
[0085] LPA receptors mediate both migration of and invasion by
pancreatic cancer cell lines: Ki16425 and LPA.sub.1-specific siRNA
effectively blocked in vitro migration in response to LPA and
peritoneal fluid (ascites) from pancreatic cancer patients; in
addition, Ki16425 blocked the LPA-induced and ascites-induced
invasion activity of a highly peritoneal metastatic pancreatic
cancer cell line (Yamada et al, J. Biol. Chem., 279, 6595-6605,
2004).
[0086] Colorectal carcinoma cell lines show significant expression
of LPA.sub.1 mRNA and respond to LPA by cell migration and
production of angiogenic factors. Overexpression of LPA receptors
has a role in the pathogenesis of thyroid cancer. LPA.sub.3 was
originally cloned from prostate cancer cells, concordant with the
ability of LPA to induce autocrine proliferation of prostate cancer
cells.
[0087] LPA has stimulatory roles in cancer progression in many
types of cancer. LPA is produced from and induces proliferation of
prostate cancer cell lines. LPA induces human colon carcinoma DLD1
cell proliferation, migration, adhesion, and secretion of
angiogenic factors through LPA.sub.1 signalling. In other human
colon carcinoma cells lines (HT29 and WiDR), LPA enhances cell
proliferation and secretion of angiogenic factors. In other colon
cancer cell lines, LPA.sub.2 and LPA.sub.3 receptor activation
results in proliferation of the cells. LPA.sub.1 is implicated in
bone metastasis and Ki16425 has been shown to inhibit metastasis to
bone in vivo (Boucharaba et al., Proc. Natl. Acad. Sci. USA, 103,
9643-9648, 2006).
[0088] In one aspect, a compound of Formula (I) is used in the
treatment of cancer. In one aspect, compounds of Formula (I) are
used in the treatment of malignant and benign proliferative
disease. In one aspect, compounds of Formula (I) are used to
prevent or reduce proliferation of tumor cells, invasion and
metastasis of carcinomas, pleural mesothelioma or peritoneal
mesothelioma, cancer pain, bone metastases. In one aspect is a
method of treating cancer in a mammal, the method comprising
administering to the mammal a compound of Formula (I) and a second
therapeutic agent, wherein the second therapeutic agent is an
anti-cancer agent. In some embodiments, radiation therapy is also
used.
[0089] The types of cancer include, but is not limited to, solid
tumors (such as those of the bladder, bowel, brain, breast,
endometrium, heart, kidney, lung, lymphatic tissue (lymphoma),
ovary, pancreas or other endocrine organ (thyroid), prostate, skin
(melanoma or basal cell cancer) or hematological tumors (such as
the leukemias) at any stage of the disease with or without
metastases.
[0090] In one aspect, LPA is a contributor to the pathogenesis of
respiratory diseases. Proinflammatory effects of LPA include
degranulation of mast cells, contraction of smooth-muscle cells and
release of cytokines from dendritic cells. LPA induces the
secretion of IL-8 from human bronchial epithelial cells. IL-8 is
found in increased concentrations in BAL fluids from patients with
asthma, chronic obstructive lung disease, pulmonary sarcoidosis and
acute respiratory distress syndrome and 11-8 has been shown to
exacerbate airway inflammation and airway remodeling of asthmatics.
LPA1, LPA2 and LPA3 receptors have all been shown to contribute to
the LPA-induced IL-8 production.
[0091] Administration of LPA in vivo induces airway
hyper-responsiveness, itch-scratch responses, infiltration and
activation of eosinophils and neutrophils, vascular remodeling, and
nociceptive flexor responses. LPA also induces histamine release
from mouse and rat mast cells. In one aspect, the effects of LPA
are mediated through LPA.sub.1 and/or LPA.sub.3. In one aspect,
compounds of Formula (I) are used in the treatment of various
allergic disorders in a mammal. In one aspect, compounds of Formula
(I) are used in the treatment of respiratory diseases, disorders or
conditions in a mammal. In one aspect, compounds of Formula (I) are
used in the treatment of asthma in a mammal. In one aspect,
compounds of Formula (I) are used in the treatment of chronic
asthma in a mammal.
[0092] The term "respiratory disease," as used herein, refers to
diseases affecting the organs that are involved in breathing, such
as the nose, throat, larynx, eustachian tubes, trachea, bronchi,
lungs, related muscles (e.g., diaphram and intercostals), and
nerves. Respiratory diseases include, but are not limited to,
asthma, adult respiratory distress syndrome and allergic
(extrinsic) asthma, non-allergic (intrinsic) asthma, acute severe
asthma, chronic asthma, clinical asthma, nocturnal asthma,
allergen-induced asthma, aspirin-sensitive asthma, exercise-induced
asthma, isocapnic hyperventilation, child-onset asthma, adult-onset
asthma, cough-variant asthma, occupational asthma,
steroid-resistant asthma, seasonal asthma, seasonal allergic
rhinitis, perennial allergic rhinitis, chronic obstructive
pulmonary disease, including chronic bronchitis or emphysema,
pulmonary hypertension, interstitial lung fibrosis and/or airway
inflammation and cystic fibrosis, and hypoxia.
[0093] In one aspect, presented herein is the use of compounds of
Formula (I) in the treatment or prevention of chronic obstructive
pulmonary disease in a mammal comprising administering to the
mammal at least once an effective amount of at least one compound
of Formula (I). In addition, chronic obstructive pulmonary disease
includes, but is not limited to, chronic bronchitis or emphysema,
pulmonary hypertension, interstitial lung fibrosis and/or airway
inflammation, and cystic fibrosis.
[0094] The nervous system is a major locus for LPA.sub.1
expression. In one aspect, provided is a compound of Formula (I)
for use in the treatment or prevention of a nervous system disorder
in a mammal. The term "nervous system disorder," as used herein
includes, but is not limited to, Alzheimer's Disease, cerebral
edema, cerebral ischemia, stroke, multiple sclerosis, neuropathies,
Parkinson's Disease, multiple sclerosis, retinal ischemia,
post-surgical cognitive dysfunction, migraine, peripheral
neuropathy/neuropathic pain, spinal cord injury, cerebral edema and
head injury.
[0095] Angiogenesis, the formation of new capillary networks from
pre-existing vasculature, is normally invoked in wound healing,
tissue growth and myocardial angiogenesis after ischemic injury.
Peptide growth factors and lysophospholipids control coordinated
proliferation, migration, adhesion, differentiation and assembly of
vascular endothelial cells (VECs) and surrounding vascular
smooth-muscle cells (VSMCs). In one aspect, dysregulation of the
processes mediating angiogenesis leads to atherosclerosis,
hypertension, tumor growth, rheumatoid arthritis and diabetic
retinopathy.
[0096] The specific effects of LPA are receptor-mediated.
[0097] In one aspect, compounds of Formula (I) are used to treat or
prevent cardiovascular disease in mammal, including but not limited
to: arrhythmia (atrial or ventricular or both); atherosclerosis and
its sequelae; angina; cardiac rhythm disturbances; myocardial
ischemia; myocardial infarction; cardiac or vascular aneurysm;
vasculitis, stroke; peripheral obstructive arteriopathy of a limb,
an organ, or a tissue; reperfusion injury following ischemia of the
brain, heart, kidney or other organ or tissue; endotoxic, surgical,
or traumatic shock; hypertension, valvular heart disease, heart
failure, abnormal blood pressure; shock; vasoconstriction
(including that associated with migraines); vascular abnormality,
inflammation, insufficiency limited to a single organ or
tissue.
[0098] In one aspect, provided herein are methods for preventing or
treating vasoconstriction, atherosclerosis and its sequelae
myocardial ischemia, myocardial infarction, aortic aneurysm,
vasculitis and stroke comprising administering at least once to the
mammal an effective amount of at least one compound of Formula (I)
or pharmaceutical composition or medicament which includes a
compound of Formula (I).
[0099] In one aspect, provided herein are methods for reducing
cardiac reperfusion injury following myocardial ischemia and/or
endotoxic shock comprising administering at least once to the
mammal an effective amount of at least one compound of Formula
(I).
[0100] In one aspect, provided herein are methods for reducing the
constriction of blood vessels in a mammal comprising administering
at least once to the mammal an effective amount of at least one
compound of Formula (I).
[0101] In one aspect, provided herein are methods for lowering or
preventing an increase in blood pressure of a mammal comprising
administering at least once to the mammal an effective amount of at
least one compound of Formula (I).
[0102] LPA is associated with various inflammatory/immune diseases.
In one aspect, compounds of Formula (I) are used to treat or
prevent inflammation in a mammal. In one aspect, antagonists of
LPA.sub.1 and/or LPA.sub.3 find use in the treatment or prevention
of inflammatory/immune disorders in a mammal.
[0103] Examples of inflammatory/immune disorders include psoriasis,
rheumatoid arthritis, vasculitis, inflammatory bowel disease,
dermatitis, osteoarthritis, asthma, inflammatory muscle disease,
allergic rhinitis, vaginitis, interstitial cystitis, scleroderma,
eczema, allogeneic or xenogeneic transplantation (organ, bone
marrow, stem cells and other cells and tissues) graft rejection,
graft-versus-host disease, lupus erythematosus, inflammatory
disease, type I diabetes, pulmonary fibrosis, dermatomyositis,
Sjogren's syndrome, thyroiditis (e.g., Hashimoto's and autoimmune
thyroiditis), myasthenia gravis, autoimmune hemolytic anemia,
multiple sclerosis, cystic fibrosis, chronic relapsing hepatitis,
primary biliary cirrhosis, allergic conjunctivitis and atopic
dermatitis.
Other Diseases, Disorders or Conditions
[0104] In accordance with one aspect, are methods for treating,
preventing, reversing, halting or slowing the progression of
LPA-dependent or LPA-mediated diseases or conditions once it
becomes clinically evident, or treating the symptoms associated
with or related to LPA-dependent or LPA-mediated diseases or
conditions, by administering to the mammal a compound of Formula
(I). In certain embodiments, the subject already has a
LPA-dependent or LPA-mediated disease or condition at the time of
administration, or is at risk of developing a LPA-dependent or
LPA-mediated disease or condition.
[0105] In certain aspects, are methods for preventing or treating
eosinophil and/or basophil and/or dendritic cell and/or neutrophil
and/or monocyte and/or T-cell recruitment comprising administering
at least once to the mammal an effective amount of at least one
compound of Formula (I).
[0106] In certain aspects, are methods for the treatment of
cystitis, including, e.g., interstitial cystitis, comprising
administering at least once to the mammal a therapeutically
effective amount of at least one compound of Formula (I).
[0107] In accordance with one aspect, methods described herein
include the diagnosis or determination of whether or not a patient
is suffering from a LPA-dependent or LPA-mediated disease or
condition by administering to the subject a therapeutically
effective amount of a compound of Formula (I) and determining
whether or not the patient responds to the treatment.
[0108] In one aspect provided herein are compounds of Formula (I),
pharmaceutically acceptable salts, pharmaceutically acceptable
prodrugs, and pharmaceutically acceptable solvates thereof, which
are antagonists of at least one LPA receptor (e.g. LPA.sub.1,
LPA.sub.2, LPA.sub.3) and are used to treat patients suffering from
one or more LPA-dependent or LPA-mediated conditions or diseases,
including, but not limited to, lung fibrosis, kindney fibrosis,
liver fibrosis, scarring, asthma, rhinitis, chronic obstructive
pulmonary disease, pulmonary hypertension, interstitial lung
fibrosis, arthritis, allergy, psoriasis, inflammatory bowel
disease, adult respiratory distress syndrome, myocardial
infarction, aneurysm, stroke, cancer, pain, proliferative disorders
and inflammatory conditions. In some embodiments, LPA-dependent
conditions or diseases include those wherein an absolute or
relative excess of LPA is present and/or observed.
[0109] In any of the aforementioned aspects the LPA-dependent or
LPA-mediated diseases or conditions include, but are not limited
to, organ fibrosis, asthma, allergic disorders, chronic obstructive
pulmonary disease, pulmonary hypertension, lung or pleural
fibrosis, peritoneal fibrosis, arthritis, allergy, cancer,
cardiovascular disease, ult respiratory distress syndrome,
myocardial infarction, aneurysm, stroke, and cancer.
[0110] In one aspect, compounds of Formula (I) are used to improve
the corneal sensitivity decrease caused by corneal operations such
as laser-assisted in situ keratomileusis (LASIK) or cataract
operation, corneal sensitivity decrease caused by corneal
degeneration, and dry eye symptom caused thereby.
[0111] In one aspect, presented herein is the use of compounds of
Formula (I) in the treatment or prevention of ocular inflammation
and allergic conjunctivitis, vernal keratoconjunctivitis, and
papillary conjunctivitis in a mammal comprising administering at
least once to the mammal an effective amount of at least one
compound of Formula (I).
[0112] In one aspect, presented herein is the use of compounds of
Formula (I) in the treatment or prevention of Sjogren disease or
inflammatory disease with dry eyes in a mammal comprising
administering at least once to the mammal an effective amount of at
least one compound of Formula (I).
[0113] In one aspect, LPA and LPA receptors (e.g. LPA.sub.1) are
involved in the pathogenesis of osteoarthritis. In one aspect,
presented herein is the use of compounds of Formula (I) in the
treatment or prevention of osteoarthritis in a mammal comprising
administering at least once to the mammal an effective amount of at
least one compound of Formula (I).
[0114] In one aspect, LPA receptors (e.g. LPA.sub.1, LPA.sub.3)
contribute to the pathogenesis of rheumatoid arthritis. In one
aspect, presented herein is the use of compounds of Formula (I) in
the treatment or prevention of rheumatoid arthritis in a mammal
comprising administering at least once to the mammal an effective
amount of at least one compound of Formula (I).
[0115] In one aspect, LPA receptors (e.g. LPA.sub.1) contribute to
adipogenesis. In one aspect, presented herein is the use of
compounds of Formula (I) in the promotion of adipose tissue
formation in a mammal comprising administering at least once to the
mammal an effective amount of at least one compound of Formula
(I).
Compounds
[0116] In some embodiments, provided herein is a compound having
the structure of Formula (I) or a pharmaceutically acceptable salt
thereof:
##STR00002##
wherein [0117] R.sup.1 is --CO.sub.2H, --CO.sub.2R.sup.D, --CN,
tetrazolyl, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.10,
--C(.dbd.O)NHSO.sub.2R.sup.10 or
--C(.dbd.O)NHCH.sub.2CH.sub.2SO.sub.3H; R.sup.D is H or
C.sub.1-C.sub.4alkyl; [0118] L.sup.1 is absent or
C.sub.1-C.sub.6alkylene; [0119] R.sup.3 is H, C.sub.1-C.sub.4alkyl,
C.sub.3-C.sub.6cycloalkyl, or C.sub.1-C.sub.4-fluoroalkyl; [0120]
R.sup.7 is H or C.sub.1-C.sub.4alkyl; [0121] R.sup.8 is H,
C.sub.1-C.sub.4alkyl, or C.sub.1-C.sub.4-fluoroalkyl; [0122]
R.sup.10 is a C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6-fluoroalkyl,
C.sub.3-C.sub.6cycloalkyl, or a substituted or unsubstituted
phenyl; [0123] each of R.sup.A, R.sup.B, and R.sup.C are
independently selected from H, F, Cl, Br, I, --CN, --OH,
C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.4-fluoroalkyl,
C.sub.1-C.sub.4-fluoroalkoxy, C.sub.1-C.sub.4alkoxy, and
C.sub.1-C.sub.4heteroalkyl; [0124] m is 0, 1, or 2; n is 0, 1, or
2; p is 0, 1, or 2.
[0125] For any and all of the embodiments, substituents are
selected from among from a subset of the listed alternatives. For
example, in some embodiments, R.sup.1 is --CO.sub.2H,
--CO.sub.2R.sup.D, --CN, tetrazolyl, or
--C(.dbd.O)NHSO.sub.2R.sup.10. For example, in some embodiments,
R.sup.1 is --CO.sub.2H or --CO.sub.2(R.sup.D). In some embodiments,
R.sup.D is H, --CH.sub.3, or --CH.sub.2CH.sub.3. In some
embodiments, R.sup.1 is --CO.sub.2H. In some embodiments, R.sup.1
is --C(.dbd.O)NHSO.sub.2R.sup.10. In some embodiments, R.sup.1 is a
carboxylic acid bioisostere.
[0126] In some embodiments, R.sup.3 is C.sub.1-C.sub.4alkyl. In
some embodiments, R.sup.3 is --CH.sub.3.
[0127] In some embodiments, R.sup.7 is H.
[0128] In some embodiments, R.sup.8 is H, C.sub.1-C.sub.4alkyl, or
C.sub.1-C.sub.4-fluoroalkyl. In some embodiments, R.sup.8 is H. In
some embodiments, R.sup.8 is H or C.sub.1-C.sub.4alkyl. In some
embodiments, R.sup.8 is H. In some embodiments, R.sup.8 is H,
--CH.sub.3, or --CF.sub.3. In some embodiments, R.sup.8 is
--CH.sub.3. In some embodiments, R.sup.8 is --CH.sub.2CH.sub.3.
[0129] In some embodiments, R.sup.10 is a C.sub.1-C.sub.6alkyl or a
substituted or unsubstituted phenyl. In some embodiments, R.sup.10
is a C.sub.1-C.sub.6alkyl. In some embodiments, R.sup.10 is a
substituted or unsubstituted phenyl.
[0130] In some embodiments, each R.sup.A is independently selected
from H, F, Cl, Br, I, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3. In some embodiments, each R.sup.A is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3. In some embodiments, each R.sup.A is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, and --OH. In some
embodiments,
[0131] each R.sup.A is independently selected from H, F, Cl,
--CH.sub.3, and --OH. In some embodiments, each R.sup.A is H.
[0132] In some embodiments, each R.sup.B is independently selected
from H, F, Cl, Br, I, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3. In some embodiments, each R.sup.B is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, and --OH. In some
embodiments, each R.sup.B is independently selected from H, F, Cl,
--CH.sub.3, and --OH. In some embodiments, each R.sup.B is H.
[0133] In some embodiments, each R.sup.C is independently selected
from H, F, Cl, Br, I, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3. In some embodiments, each R.sup.C is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3. In some embodiments, each R.sup.C is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, and --OH. In some
embodiments, each R.sup.C is independently selected from H, F, Cl,
and --OH. In some embodiments, each R.sup.C is independently
selected from H, F, and Cl. In some embodiments, each R.sup.C is
H.
[0134] In some embodiments, m is 0 or 1. In some embodiments, m is
0. In some embodiments, m is 1. In some embodiments, p is 0 or 1.
In some embodiments, p is 0. In some embodiments, p is 1. In some
embodiments, n is 0, 1, or 2. In some embodiments, n is 0 or 1. In
some embodiments, n is 0. In some embodiments, n is 1. In some
embodiments, n is 2.
[0135] In some embodiments, R.sup.1 is --CO.sub.2H,
--CO.sub.2R.sup.D or --C(.dbd.O)NHSO.sub.2R.sup.10; R.sup.3 is
C.sub.1-C.sub.4alkyl; R.sup.7 is H; R.sup.10 is a
C.sub.1-C.sub.6alkyl or a substituted or unsubstituted phenyl; each
R.sup.A is independently selected from H, F, Cl, Br, I, --CH.sub.3,
--CF.sub.3, --OH, --OCF.sub.3, and --OCH.sub.3; each R.sup.B is
independently selected from H, F, Cl, Br, I, --CH.sub.3,
--CF.sub.3, --OH, --OCF.sub.3, and --OCH.sub.3; each R.sup.C is
independently selected from H, F, Cl, Br, I, --CH.sub.3,
--CF.sub.3, --OH, --OCF.sub.3, and --OCH.sub.3; m is 0 or 1; p is 0
or 1.
[0136] In some embodiments, R.sup.1 is --CO.sub.2H or
--CO.sub.2R.sup.D; R.sup.3 is --CH.sub.3 or --CH.sub.2CH.sub.3;
R.sup.8 is H, --CH.sub.3 or --CF.sub.3; R.sup.D is H, --CH.sub.3,
or --CH.sub.2CH.sub.3.
[0137] In some embodiments, R.sup.1 is --CO.sub.2H; R.sup.3 is
--CH.sub.3; R.sup.8 is --CH.sub.3; L.sup.1 is absent, --CH.sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2CH.sub.2CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH(CH.sub.2CH.sub.3)--,
--C(CH.sub.2CH.sub.3).sub.2--, --CH.sub.2CH(CH.sub.3)--, or
--CH.sub.2C(CH.sub.3).sub.2--. In some embodiments, R.sup.1 is
--CO.sub.2H; R.sup.8 is --CH.sub.3; L.sup.1 is --CH.sub.2--. In
some embodiments, R.sup.1 is --CO.sub.2H; R.sup.8 is --CH.sub.3;
L.sup.1 is --CH.sub.2--; m is 0; n is 0 or 1; p is 0.
[0138] In some embodiments, the compound of Formula (I) has the
following structure:
##STR00003##
[0139] In some embodiments, L.sup.1 is absent, --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH(CH.sub.2CH.sub.3)--,
or --C(CH.sub.2CH.sub.3).sub.2--; each R.sup.C is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3; m is 0; n is 0, 1, or 2; p is 0.
[0140] In some embodiments, L.sup.1 is absent or --CH.sub.2--; each
R.sup.C is independently selected from H, F, Cl, --CH.sub.3,
--CF.sub.3, and --OH; n is 0 or 1.
[0141] In some embodiments, the compound of Formula (I) has the
following structure:
##STR00004##
[0142] In some embodiments, L.sup.1 is absent, --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH(CH.sub.2CH.sub.3)--,
or --C(CH.sub.2CH.sub.3).sub.2--; each R.sup.C is independently
selected from H, F, Cl, --CH.sub.3, --CF.sub.3, --OH, --OCF.sub.3,
and --OCH.sub.3; m is 0; n is 0, 1, or 2; p is 0.
[0143] In some embodiments, L.sup.1 is absent or --CH.sub.2--; each
R.sup.C is independently selected from H, F, Cl, --CH.sub.3,
--CF.sub.3, and --OH; n is 0 or 1. In some embodiments, L.sup.1 is
--CH.sub.2--.
[0144] In some embodiments, R.sup.1 is
--C(.dbd.O)NHSO.sub.2R.sup.10; R.sup.3 is --CH.sub.3 or
--CH.sub.2CH.sub.3; R.sup.8 is H, --CH.sub.3 or --CF.sub.3;
R.sup.10 is --CH.sub.3, or --CH.sub.2CH.sub.3.
[0145] In some embodiments, L.sup.1 is as described in Table 1
and/or Table 2.
[0146] In some embodiments,
##STR00005##
as defined in Table 1.
[0147] In some embodiments,
##STR00006##
as defined in Table 1.
[0148] In some embodiments,
##STR00007##
[0149] In some embodiments,
##STR00008##
[0150] In some embodiments, the compound of Formula (I) has a
structure selected from:
##STR00009##
[0151] Any combination of the groups described above for the
various variables is contemplated herein. Throughout the
specification, groups and substituents thereof are chosen by one
skilled in the field to provide stable moieties and compounds.
[0152] In some embodiments, compounds of Formula (I) include, but
are not limited to, those described in Table 1, Table 2, Table 3
and FIGS. 1 to 7.
TABLE-US-00001 TABLE 1 ##STR00010## Cmpd # R.sup.A B -L.sup.1-
R.sup.D C R.sup.8 M + H* 1 H Phen-1,4- --CH.sub.2-- H
2-Trifluoromethyl- CH.sub.3 525 ylene phenyl 2 H Phen-1,4-
--CH.sub.2-- H 3-Trifluoromethyl- CH.sub.3 525 ylene phenyl 3 H
Phen-1,4- --CH.sub.2-- H 2,4-Dichloro-phenyl CH.sub.3 525 ylene 4 H
Phen-1,4- --CH.sub.2-- H 2-Fluoro-phenyl CH.sub.3 475 ylene 5 H
Phen-1,4- --CH.sub.2-- H 3-Bromo-phenyl CH.sub.3 535 ylene 6 H
Phen-1,4- --CH.sub.2-- H 2-Methoxy-phenyl CH.sub.3 487 ylene 7 H 2-
--CH.sub.2-- H 2-Chloro-phenyl CH.sub.3 521 Methoxyphen- 1,5-ylene
8 H Phen-1,4- -- H 2-Chloro-phenyl CH.sub.3 477 ylene 9 H Phen-1,2-
-- H 2-Chloro-phenyl CH.sub.3 477 ylene 10 H Phen-1,2- --CH.sub.2--
H 2-Chloro-phenyl CH.sub.3 491 ylene 11 H Phen-1,4- --CH.sub.2-- H
2-Chloro-phenyl CH.sub.3 491 ylene 12 H Phen-1,3- --CH.sub.2-- H
2-Chloro-phenyl CH.sub.3 491 ylene 13 H Phen-1,4-
--CH.sub.2CH.sub.2-- H 2-Chloro-phenyl CH.sub.3 505 ylene 14 H 2-
--CH.sub.2-- H 2-Chloro-phenyl CH.sub.3 509 Fluorophen- 1,5-ylene
15 H 4- --CH.sub.2-- H 2-Chloro-phenyl CH.sub.3 509 Fluorophen-
1,5-ylene 16 H Phen-1,4- --CH.sub.2-- --CH.sub.3 2-Chloro-phenyl
CH.sub.3 505 ylene 17 H Phen-1,4- --CH(CH.sub.3)--
--CH.sub.2CH.sub.3 2-Chloro-phenyl CH.sub.3 533 ylene 18 H
Phen-1,4- --CH(CH.sub.3)-- H 2-Chloro-phenyl CH.sub.3 505 ylene 19
H Phen-1,4- --C(CH.sub.3).sub.2-- H 2-Chloro-phenyl CH.sub.3 519
ylene 20 H Phen-1,4- --CH(CH.sub.3)-- H 2-Fluoro-phenyl CH.sub.3
489 ylene 21 H Phen-1,4- --CH.sub.2CH.sub.2CH.sub.2-- H
2-Chloro-phenyl CH.sub.3 519 ylene 22 H Phen-1,3- -- H
2-Chloro-phenyl CH.sub.3 477 ylene 23 H Phen-1,4- --CH.sub.2-- H
2-Fluoro-4-chloro- CH.sub.3 509 ylene phenyl 24 H Phen-1,4-
--CH.sub.2-- H 2-Fluoro-phenyl (R)--CH.sub.3 475 ylene 25
--CH.sub.3 Phen-1,4- --CH.sub.2-- H 2-Fluoro-phenyl (R)--CH.sub.3
489 ylene 26 H Phen-1,4- --C(CH.sub.3).sub.2-- H 2-Fluoro-phenyl
CH.sub.3 503 ylene 27 H Phen-1,4- --CH.sub.2-- H 2-Chloro-phenyl
(R)--CH.sub.3 491 ylene 28 H Phen-1,4- --C(CH.sub.3).sub.2-- H
2-Chloro-phenyl (R)--CH.sub.3 519 ylene 29 H Phen-1,4-
--C(CH.sub.3).sub.2-- H 2-Fluoro-phenyl (R)--CH.sub.3 503 ylene 30
H Phen-1,4- --CH(CH.sub.3)-- H 2-Fluoro-phenyl (R)--CH.sub.3 489
ylene 31 H Phen-1,4- --CH(CH.sub.3)-- H 2-Chloro-phenyl
(R)--CH.sub.3 505 ylene 32 H Phen-1,4- --CH.sub.2-- H
2,6-Dichloro-phenyl CH.sub.3 525 ylene 33 --CH.sub.3 Phen-1,4-
--CH(CH.sub.3)-- H 2-Fluoro-phenyl (R)--CH.sub.3 503 ylene 34 H
Phen-1,4- --CH.sub.2-- H 2-Fluoro-phenyl (S)--CH.sub.3 475 ylene 35
H Phen-1,4- --CH.sub.2-- H 2-Chloro-phenyl (S)--CH.sub.3 491 ylene
36 H Phen-1,4- --CH.sub.2-- H 2-Chloro-phenyl H 477 ylene 37 H
Phen-1,4- --CH.sub.2-- H Phenyl (R)--CH.sub.3 457 ylene 38 H
Phen-1,4- --CH.sub.2-- H 2,3-Difluoro-phenyl CH.sub.3 493 ylene 39
H Phen-1,4- --CH.sub.2-- H 2,4-Difluoro-phenyl CH.sub.3 493 ylene
40 H Phen-1,4- --CH.sub.2-- H 2-Fluoro-4- CH.sub.3 505 ylene
methoxy-phenyl 41 H Phen-1,4- --CH.sub.2-- H 2,5-Difluoro-phenyl
CH.sub.3 493 ylene 42 H Phen-1,4- --CH.sub.2-- H
2,6-Difluoro-phenyl CH.sub.3 493 ylene 43 H Phen-1,3- --CH.sub.2--
H Phenyl (R)--CH.sub.3 457 ylene 44 H Phen-1,4- -- H Phenyl
(R)--CH.sub.3 443 ylene 45 H Phen-1,2- --CH.sub.2-- H Phenyl
(R)--CH.sub.3 457 ylene 46 H Phen-1,4- --CH.sub.2-- H
2-Methyl-phenyl (R)--CH.sub.3 471 ylene 47 H Phen-1,4-
--CH(CH.sub.3)-- H 2-Chloro-phenyl (R)--CH.sub.3 505 ylene 48 H
Phen-1,4- --CH(CH.sub.3)-- H 2-Chloro-phenyl (R)--CH.sub.3 505
ylene 49 H Phen-1,4- --CH.sub.2-- H 2-Chloro-phenyl (R)--CH.sub.3
525 ylene 50 H Phen-1,4- --CH(CH.sub.2CH.sub.3)-- H 2-Chloro-phenyl
(R)--CH.sub.3 519 ylene 51 Cl Phen-1,4- --CH.sub.2-- H
2-Chloro-phenyl (R)--CH.sub.3 526 ylene 52 F Phen-1,4- --CH.sub.2--
H 2-Chloro-phenyl (R)--CH.sub.3 509 ylene 53 H Phen-1,4- -- H
2-Chloro-phenyl (R)--CH.sub.3 477 ylene 56 H Phen-1,4- --CH.sub.2--
H 3,5-Dibromo-phenyl (R)--CH.sub.3 615 ylene 57 H Phen-1,4-
--CH.sub.2-- H Phenyl (S)--CH.sub.3 457 ylene 58 H Phen-1,4-
--CH.sub.2-- H 3-Hydroxy-phenyl (R)--CH.sub.3 473 ylene 59 H
Phen-1,4- --CH.sub.2-- H Phenyl --CH.sub.3 457 ylene 60 H Phen-1,4-
--CH.sub.2-- H Phenyl-d5 --CD.sub.3 466 ylene *mass spectrometric
data
TABLE-US-00002 TABLE 2 ##STR00011## Cmpd # L.sup.1 R.sup.1 R.sup.C
R.sup.8 M + H* 61 --CH.sub.2-- --CN Cl (R)--CH.sub.3 472 62
--CH.sub.2-- --CN F (R)--CH.sub.3 456 63 --CH.sub.2--
2H-Tetrazol-5-yl F (R)--CH.sub.3 499 64 --CH.sub.2--
2H-Tetrazol-5-yl Cl (R)--CH.sub.3 515 65 --CH.sub.2--
--C(.dbd.NH)--NH.sub.2 F (R)--CH.sub.3 473 66 --CH.sub.2--
--C(.dbd.NH)--NH--Ac F (R)--CH.sub.3 515 67 --CH.sub.2--
--C(.dbd.O)--NHCH.sub.2CH.sub.2 SO.sub.3H H (R)--CH.sub.3 564 *mass
spectrometric data
TABLE-US-00003 TABLE 3 ##STR00012## Cmpd # R.sup.1 R.sup.C R.sup.8
67 --C(.dbd.O)NHSO.sub.2Me H (R)--CH.sub.3 68
--C(.dbd.O)NHSO.sub.2Me H H 69 --C(.dbd.O)NHSO.sub.2Me 2-F
(R)--CH.sub.3 70 --C(.dbd.O)NHSO.sub.2Me 2-F H 71
--C(.dbd.O)NHSO.sub.2Me 2-Cl (R)--CH.sub.3 72
--C(.dbd.O)NHSO.sub.2Me 2-Cl H 73 --C(.dbd.O)NHSO.sub.2Me 2-
CH.sub.3 (R)--CH.sub.3 74 --C(.dbd.O)NHSO.sub.2Me 2- CH.sub.3 H 75
--C(.dbd.O)NHSO.sub.2Me 2-CF.sub.3 (R)--CH.sub.3 76
--C(.dbd.O)NHSO.sub.2Me 2-CF.sub.3 H 77 --C(.dbd.O)NHSO.sub.2Me 3-F
(R)--CH.sub.3 78 --C(.dbd.O)NHSO.sub.2Me 3-F H 79
--C(.dbd.O)NHSO.sub.2Me 3-Cl (R)--CH.sub.3 80
--C(.dbd.O)NHSO.sub.2Me 3-Cl H 81 --C(.dbd.O)NHSO.sub.2Me 3-
CH.sub.3 (R)--CH.sub.3 82 --C(.dbd.O)NHSO.sub.2Me 3- CH.sub.3 H 83
--C(.dbd.O)NHSO.sub.2Me 3-CF.sub.3 (R)--CH.sub.3 84
--C(.dbd.O)NHSO.sub.2Me 3-CF.sub.3 H 85 --C(.dbd.O)NHSO.sub.2Ph H
(R)--CH.sub.3 86 --C(.dbd.O)NHSO.sub.2Ph H H 87
--C(.dbd.O)NHSO.sub.2Ph 2-F (R)--CH.sub.3 88
--C(.dbd.O)NHSO.sub.2Ph 2-F H 89 --C(.dbd.O)NHSO.sub.2Ph 2-Cl
(R)--CH.sub.3 90 --C(.dbd.O)NHSO.sub.2Ph 2-Cl H 91
--C(.dbd.O)NHSO.sub.2Ph 2- CH.sub.3 (R)--CH.sub.3 92
--C(.dbd.O)NHSO.sub.2Ph 2- CH.sub.3 H 93 --C(.dbd.O)NHSO.sub.2Ph
2-CF.sub.3 (R)--CH.sub.3 94 --C(.dbd.O)NHSO.sub.2Ph 2-CF.sub.3 H 95
--C(.dbd.O)NHSO.sub.2Ph 3-F (R)--CH.sub.3 96
--C(.dbd.O)NHSO.sub.2Ph 3-F H 97 --C(.dbd.O)NHSO.sub.2Ph 3-Cl
(R)--CH.sub.3 98 --C(.dbd.O)NHSO.sub.2Ph 3-Cl H 99
--C(.dbd.O)NHSO.sub.2Ph 3- CH.sub.3 (R)--CH.sub.3 100
--C(.dbd.O)NHSO.sub.2Ph 3- CH.sub.3 H 101 --C(.dbd.O)NHSO.sub.2Ph
3-CF.sub.3 (R)--CH.sub.3 102 --C(.dbd.O)NHSO.sub.2Ph 3-CF.sub.3
H
Synthesis of Compounds
[0153] Compounds of Formula (I) described herein are synthesized
using standard synthetic techniques or using methods known in the
art in combination with methods described herein. In additions,
solvents, temperatures and other reaction conditions presented
herein may vary.
[0154] The starting material used for the synthesis of the
compounds of Formula (I) are either synthesized or obtained from
commercial sources, such as, but not limited to, Sigma-Aldrich,
Fluka, Acros Organics, Alfa Aesar, and the like. General methods
for the preparation of compounds can be modified by the use of
appropriate reagents and conditions for the introduction of the
various moieties found in the formulae as provided herein.
[0155] In some embodiments, the compounds of Formula (I) are
prepared as outlined herein.
Further Forms of Compounds
[0156] In one aspect, compounds of Formula (I) possess one or more
stereocenters and each stereocenter exists independently in either
the R or S configuration. The compounds presented herein include
all diastereomeric, and enantiomeric forms. Stereoisomers are
obtained, if desired, by methods such as, stereoselective synthesis
and/or the separation of stereoisomers by chiral chromatographic
columns.
[0157] The methods and formulations described herein include the
use of N-oxides (if appropriate), crystalline forms (also known as
polymorphs), amorphous phases, and/or pharmaceutically acceptable
salts of compounds having the structure of Formula (I), as well as
metabolites and active metabolites of these compounds having the
same type of activity. In some situations, compounds may exist as
tautomers. All tautomers are included within the scope of the
compounds presented herein. In specific embodiments, the compounds
described herein exist in solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like. In other
embodiments, the compounds described herein exist in unsolvated
form.
[0158] In some embodiments, compounds described herein are prepared
as prodrugs. A "prodrug" refers to an agent that is converted into
the parent drug in vivo. In certain embodiments, upon in vivo
administration, a prodrug is chemically converted to the
biologically, pharmaceutically or therapeutically active form of
the compound. In certain embodiments, a prodrug is enzymatically
metabolized by one or more steps or processes to the biologically,
pharmaceutically or therapeutically active form of the
compound.
[0159] In some embodiments, sites on the aromatic ring portion of
compounds of Formula (I) are susceptible to various metabolic
reactions. Incorporation of appropriate substituents on the
aromatic ring structures will reduce, minimize or eliminate this
metabolic pathway. In specific embodiments, the appropriate
substituent to decrease or eliminate the susceptibility of the
aromatic ring to metabolic reactions is, by way of example only, a
deuterium, a halogen, or an alkyl group.
[0160] In another embodiment, the compounds described herein are
labeled isotopically or by another other means, including, but not
limited to, the use of chromophores or fluorescent moieties,
bioluminescent labels, or chemiluminescent labels.
[0161] In one aspect, substitution with isotopes such as deuterium
affords certain therapeutic advantages resulting from greater
metabolic stability, such as, for example, increased in vivo
half-life or reduced dosage requirements.
[0162] "Pharmaceutically acceptable," as used herein, refers a
material, such as a carrier or diluent, which does not abrogate the
biological activity or properties of the compound, and is
relatively nontoxic, i.e., the material may be administered to an
individual without causing undesirable biological effects or
interacting in a deleterious manner with any of the components of
the composition in which it is contained.
[0163] In some embodiments, pharmaceutically acceptable salts are
obtained by reacting a compound of Formula (I) with acids.
Pharmaceutically acceptable salts are also obtained by reacting a
compound of Formula (I) with a base to form a salt.
[0164] Compounds described herein may be formed as, and/or used as,
pharmaceutically acceptable salts. The type of pharmaceutical
acceptable salts, include, but are not limited to: (1) acid
addition salts, formed by reacting the free base form of the
compound with a pharmaceutically acceptable inorganic acid (e.g.
hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric
acid, and the like); or with an organic acid (e.g. acetic acid,
propionic acid, glycolic acid, pyruvic acid, lactic acid, malonic
acid, succinic acid, malic acid, maleic acid, fumaric acid,
trifluoroacetic acid, tartaric acid, citric acid, benzoic acid,
cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic
acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid,
benzenesulfonic acid, toluenesulfonic acid, 2-naphthalenesulfonic
acid, salicylic acid, stearic acid, muconic acid, butyric acid,
phenylacetic acid, phenylbutyric acid, valproic acid, and the
like); (2) salts formed when an acidic proton present in the parent
compound is replaced by a metal ion, e.g., an alkali metal ion
(e.g. lithium, sodium, potassium), an alkaline earth ion (e.g.
magnesium, or calcium), or an aluminum ion. In some cases,
compounds described herein may coordinate with an organic base,
such as, but not limited to, ethanolamine, diethanolamine,
triethanolamine, tromethamine, N-methylglucamine,
dicyclohexylamine, tris(hydroxymethyl)methylamine. In other cases,
compounds described herein may form salts with amino acids such as,
but not limited to, arginine, lysine, and the like. Acceptable
inorganic bases used to form salts with compounds that include an
acidic proton, include, but are not limited to, aluminum hydroxide,
calcium hydroxide, potassium hydroxide, sodium carbonate, sodium
hydroxide, and the like. In some embodiments, a sodium salt of the
compound of Formula (I) is prepared.
[0165] It should be understood that a reference to a
pharmaceutically acceptable salt includes the solvent addition
forms or crystal forms thereof, particularly solvates or
polymorphs. Solvates contain either stoichiometric or
non-stoichiometric amounts of a solvent, and may be formed during
the process of crystallization with pharmaceutically acceptable
solvents such as water, ethanol, and the like. Hydrates are formed
when the solvent is water, or alcoholates are formed when the
solvent is alcohol. In addition, the compounds provided herein can
exist in unsolvated as well as solvated forms. In general, the
solvated forms are considered equivalent to the unsolvated forms
for the purposes of the compounds and methods provided herein.
[0166] Compounds described herein, such as compounds of Formula
(I), may be in various forms, including but not limited to,
amorphous forms, milled forms and nano-particulate forms. In
addition, compounds described herein include crystalline forms,
also known as polymorphs. Polymorphs include the different crystal
packing arrangements of the same elemental composition of a
compound.
Certain Terminology
[0167] Unless otherwise stated, the following terms used in this
application, including the specification and claims, have the
definitions given below. It must be noted that, as used in the
specification and the appended claims, the singular forms "a," "an"
and "the" include plural referents unless the context clearly
dictates otherwise. Unless otherwise indicated, conventional
methods of mass spectroscopy, NMR, HPLC, protein chemistry,
biochemistry, recombinant DNA techniques and pharmacology are
employed. In this application, the use of "or" or "and" means
"and/or" unless stated otherwise. Furthermore, use of the term
"including" as well as other forms, such as "include", "includes,"
and "included," is not limiting. The section headings used herein
are for organizational purposes only and are not to be construed as
limiting the subject matter described.
[0168] An "alkyl" refers to an aliphatic hydrocarbon. The alkyl may
be saturated or unsaturated. The alkyl, whether saturated or
unsaturated, is a branched alkyl or straight chain alkyl. Typical
alkyl groups include, but are in no way limited to, methyl, ethyl,
propyl, isopropyl, butyl, isobutyl, sec-butyl, tertiary butyl,
pentyl, neopentyl, hexyl, allyl, but-2-enyl, but-3-enyl, and the
like.
[0169] The term "alkylene" refers to a divalent alkyl radical.
Typical alkylene groups include, but are not limited to,
--CH.sub.2--, --CH(CH.sub.3)--, --C(CH.sub.3).sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2CH(CH.sub.3)--,
--CH.sub.2C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2CH.sub.2--,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--, and the like.
[0170] An "alkoxy" group refers to a (alkyl)O-- group, where alkyl
is as defined herein.
[0171] "Cycloalkyl" refers to cyclopropyl, cyclobutyl, cyclopentyl,
cyclopentenyl, cyclohexyl, and cyclohexenyl.
[0172] The term "halo" or, alternatively, "halogen" or "halide"
means fluoro, chloro, bromo or iodo.
[0173] The term "fluoroalkyl" refers to an alkyl in which one or
more hydrogen atoms are replaced by a fluorine atom.
[0174] The term "heteroalkyl" refers to an alkyl group in which one
or more skeletal atoms of the alkyl are selected from an atom other
than carbon, e.g., oxygen, nitrogen (e.g. NH or Nalkyl), sulfur, or
combinations thereof. In some embodiments, one aspect, heteroalkyl
refers to an alkyl group in which one of the skeletal atoms of the
alkyl is oxygen.
[0175] The term "optionally substituted" or "substituted" means
that the referenced group may be substituted with one or more
additional group(s) individually and independently selected from
halogen, --CN, --NH.sub.2, --OH, --NH(CH.sub.3),
--N(CH.sub.3).sub.2, --CO.sub.2H, --CO.sub.2alkyl,
--C(.dbd.O)NH.sub.2, --C(.dbd.O)NHalkyl, --C(.dbd.O)N(alkyl).sub.2,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NH(alkyl),
--S(.dbd.O).sub.2N(alkyl).sub.2, alkyl, cycloalkyl, fluoroalkyl,
heteroalkyl, alkoxy, fluoroalkoxy, --S-- alkyl, or
--S(.dbd.O).sub.2alkyl. In some embodiments, an optional
substituent is selected from halogen, --CN, --NH.sub.2, --OH,
--NH(CH.sub.3), --N(CH.sub.3).sub.2, --CH.sub.3,
--CH.sub.2CH.sub.3, --CF.sub.3, --OCH.sub.3, --OCH.sub.2CH.sub.3,
and --OCF.sub.3. In some embodiments, substituted groups are
substituted with one or two of the preceding groups. In some
embodiments, substituted groups are substituted with one of the
preceding groups.
[0176] The term "acceptable" with respect to a formulation,
composition or ingredient, as used herein, means having no
persistent detrimental effect on the general health of the subject
being treated.
[0177] The term "modulate," as used herein, means to interact with
a target either directly or indirectly so as to alter the activity
of the target, including, by way of example only, to enhance the
activity of the target, to inhibit the activity of the target, to
limit the activity of the target, or to extend the activity of the
target.
[0178] The term "modulator," as used herein, refers to a molecule
that interacts with a target either directly or indirectly. The
interactions include, but are not limited to, the interactions of
an agonist, partial agonist, an inverse agonist and antagonist. In
one embodiment, a modulator is an antagonist.
[0179] The term "agonist," as used herein, refers to a molecule
such as a compound, a drug, an enzyme activator or a hormone
modulator that binds to a specific receptor and triggers a response
in the cell. An agonist mimics the action of an endogenous ligand
(such as LPA, prostaglandin, hormone or neurotransmitter) that
binds to the same receptor.
[0180] The term "antagonist," as used herein, refers to a molecule
such as a compound, which diminishes, inhibits, or prevents the
action of another molecule or the activity of a receptor site.
Antagonists include, but are not limited to, competitive
antagonists, non-competitive antagonists, uncompetitive
antagonists, partial agonists and inverse agonists.
[0181] The term "LPA-dependent", as used herein, refers to
conditions or disorders that would not occur, or would not occur to
the same extent, in the absence of LPA.
[0182] The term "LPA-mediated", as used herein, refers to refers to
conditions or disorders that might occur in the absence of LPA but
can occur in the presence of LPA.
[0183] "Selectivity" for one LPA receptor versus other LPA
receptors means that the compound has an IC.sub.50 (Ca Flux assay)
for the indicated LPA receptor that is at least 10-fold less than
the IC.sub.50 for other LPA receptors. In some embodiments,
selectivity for one LPA receptor versus other LPA receptor means
that the compound has an IC.sub.50 for the indicated LPA receptor
that is at least 10-fold, at least 20-fold, at least 40-fold, at
least 50-fold, at least 100-fold, at least 200-fold, at least
500-fold, or at least 1000-fold, less than the IC.sub.50 for other
LPA receptors. For example, a selective LPA.sub.1 receptor
antagonist has an IC.sub.50 that is at least 10-fold, at least
20-fold, at least 40-fold, at least 50-fold, at least 100-fold, at
least 200-fold, at least 500-fold, or at least 1000-fold, less than
the IC.sub.50 for other LPA receptors (e.g. LPA.sub.2,
LPA.sub.3).
[0184] The terms "co-administration" or the like, as used herein,
are meant to encompass administration of the selected therapeutic
agents to a single patient, and are intended to include treatment
regimens in which the agents are administered by the same or
different route of administration or at the same or different
time.
[0185] The terms "effective amount" or "therapeutically effective
amount," as used herein, refer to a sufficient amount of an agent
or a compound being administered which will relieve to some extent
one or more of the symptoms of the disease or condition being
treated. The result can be reduction and/or alleviation of the
signs, symptoms, or causes of a disease, or any other desired
alteration of a biological system. For example, an "effective
amount" for therapeutic uses is the amount of the composition
comprising a compound as disclosed herein required to provide a
clinically significant decrease in disease symptoms. An appropriate
"effective" amount in any individual case may be determined using
techniques, such as a dose escalation study.
[0186] The term "pharmaceutical combination" as used herein, means
a product that results from the mixing or combining of more than
one active ingredient and includes both fixed and non-fixed
combinations of the active ingredients. The term "fixed
combination" means that the active ingredients, e.g. a compound of
Formula (I) and a co-agent, are both administered to a patient
simultaneously in the form of a single entity or dosage. The term
"non-fixed combination" means that the active ingredients, e.g. a
compound of Formula (I) and a co-agent, are administered to a
patient as separate entities either simultaneously, concurrently or
sequentially with no specific intervening time limits, wherein such
administration provides effective levels of the two compounds in
the body of the patient. The latter also applies to cocktail
therapy, e.g. the administration of three or more active
ingredients.
[0187] The term "subject" or "patient" encompasses mammals.
Examples of mammals include, but are not limited to, humans,
chimpanzees, apes, monkey, cattle, horses, sheep, goats, swine,
rabbits, dogs, cats, rodents, rats, mice guinea pigs, and the like.
In one embodiment, the mammal is a human.
[0188] The terms "treat," "treating" or "treatment," as used
herein, include alleviating, abating or ameliorating at least one
symptom of a disease disease or condition, preventing additional
symptoms, inhibiting the disease or condition, e.g., arresting the
development of the disease or condition, relieving the disease or
condition, causing regression of the disease or condition,
relieving a condition caused by the disease or condition, or
stopping the symptoms of the disease or condition either
prophylactically and/or therapeutically.
Pharmaceutical Compositions/Formulations and Routes of
Administration
[0189] In some embodiments, the compounds described herein are
formulated into pharmaceutical compositions. Pharmaceutical
compositions are formulated in a conventional manner using one or
more pharmaceutically acceptable inactive ingredients that
facilitate processing of the active compounds into preparations
that can be used pharmaceutically. Proper formulation is dependent
upon the route of administration chosen. A summary of
pharmaceutical compositions described herein can be found, for
example, in Remington: The Science and Practice of Pharmacy,
Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover,
John E., Remington's Pharmaceutical Sciences, Mack Publishing Co.,
Easton, Pa. 1975; Liberman, H. A. and Lachman, L., Eds.,
Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980;
and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh
Ed. (Lippincott Williams & Wilkins 1999), herein incorporated
by reference for such disclosure.
[0190] A pharmaceutical composition, as used herein, refers to a
mixture of a compound of Formula (I) with other chemical components
(i.e. pharmaceutically acceptable inactive ingredients), such as
carriers, excipients, binders, filling agents, suspending agents,
flavoring agents, sweetening agents, disintegrating agents,
dispersing agents, surfactants, lubricants, colorants, diluents,
solubilizers, moistening agents, plasticizers, stabilizers,
penetration enhancers, wetting agents, anti-foaming agents,
antioxidants, preservatives, or one or more combination thereof.
The pharmaceutical composition facilitates administration of the
compound to an organism.
[0191] Pharmaceutical formulations described herein are
administerable to a subject in a variety of ways by multiple
administration routes, including but not limited to, oral,
parenteral (e.g., intravenous, subcutaneous, intramuscular,
intramedullary injections, intrathecal, direct intraventricular,
intraperitoneal, intralymphatic, intranasal injections),
intranasal, buccal, topical or transdermal administration routes.
The pharmaceutical formulations described herein include, but are
not limited to, aqueous liquid dispersions, self-emulsifying
dispersions, solid solutions, liposomal dispersions, aerosols,
solid dosage forms, powders, immediate release formulations,
controlled release formulations, fast melt formulations, tablets,
capsules, pills, delayed release formulations, extended release
formulations, pulsatile release formulations, multiparticulate
formulations, and mixed immediate and controlled release
formulations.
[0192] In some embodiments, the compounds of Formula (I) are
administered orally.
[0193] In some embodiments, the compounds of Formula (I) are
administered topically. In such embodiments, the compound of
Formula (I) is formulated into a variety of topically administrable
compositions, such as solutions, suspensions, lotions, gels,
pastes, shampoos, scrubs, rubs, smears, medicated sticks, medicated
bandages, balms, creams or ointments. In one aspect, the compounds
of Formula (I) are administered topically to the skin.
[0194] In another aspect, the compounds of Formula (I) are
administered by inhalation.
[0195] In another aspect, the compounds of Formula (I) are
formulated for intranasal administration. Such formulations include
nasal sprays, nasal mists, and the like.
[0196] In another aspect, the compounds of Formula (I) are
formulated as eye drops.
[0197] In any of the aforementioned aspects are further embodiments
in which the effective amount of the compound of Formula (I) is:
(a) systemically administered to the mammal; and/or (b)
administered orally to the mammal; and/or (c) intravenously
administered to the mammal; and/or (d) administered by inhalation
to the mammal; and/or (e) administered by nasal administration to
the mammal; or and/or (f) administered by injection to the mammal;
and/or (g) administered topically to the mammal; and/or (h)
administered by ophthalmic administration; and/or (i) administered
rectally to the mammal; and/or (j) adminstered non-systemically or
locally to the mammal.
[0198] In any of the aforementioned aspects are further embodiments
comprising single administrations of the effective amount of the
compound, including further embodiments in which (i) the compound
is administered once; (ii) the compound is administered to the
mammal multiple times over the span of one day; (iii) continually;
or (iv) continuously.
[0199] In any of the aforementioned aspects are further embodiments
comprising multiple administrations of the effective amount of the
compound, including further embodiments in which (i) the compound
is administered continuously or intermittently: as in a a single
dose; (ii) the time between multiple administrations is every 6
hours; (iii) the compound is administered to the mammal every 8
hours; (iv) the compound is administered to the mammal every 12
hours; (v) the compound is administered to the mammal every 24
hours. In further or alternative embodiments, the method comprises
a drug holiday, wherein the administration of the compound is
temporarily suspended or the dose of the compound being
administered is temporarily reduced; at the end of the drug
holiday, dosing of the compound is resumed. In one embodiment, the
length of the drug holiday varies from 2 days to 1 year.
[0200] In certain embodiments, a compound as described herein is
administered in a local rather than systemic manner.
[0201] In some embodiments, the compound described herein is
administered topically. In some embodiments, the compound described
herein is administered systemically.
[0202] In some embodiments, the pharmaceutical formulation is in
the form of a tablet. In other embodiments, pharmaceutical
formulations of the compounds of Formula (I) are in the form of a
capsule.
[0203] In one aspect, liquid formulation dosage forms for oral
administration are in the form of aqueous suspensions or solutions
selected from the group including, but not limited to, aqueous oral
dispersions, emulsions, solutions, elixirs, gels, and syrups.
[0204] For administration by inhalation, a compound of Formula (I)
is formulated for use as an aerosol, a mist or a powder.
[0205] For buccal or sublingual administration, the compositions
may take the form of tablets, lozenges, or gels formulated in a
conventional manner.
[0206] In some embodiments, compounds of Formula (I) are prepared
as transdermal dosage forms.
[0207] In one aspect, a compound of Formula (I) is formulated into
a pharmaceutical composition suitable for intramuscular,
subcutaneous, or intravenous injection.
[0208] In some embodiments, the compounds described herein may be
administered topically and can be formulated into a variety of
topically administrable compositions, such as solutions,
suspensions, lotions, gels, pastes, medicated sticks, balms, creams
or ointments.
[0209] In some embodiments, the compounds of Formula (I) are
formulated in rectal compositions such as enemas, rectal gels,
rectal foams, rectal aerosols, suppositories, jelly suppositories,
or retention enemas.
Methods of Dosing and Treatment Regimens
[0210] In one embodiment, the compounds of Formula (I) are used in
the preparation of medicaments for the treatment of LPA-dependent
or LPA-mediated diseases or conditions. In addition, a method for
treating any of the diseases or conditions described herein in a
subject in need of such treatment, involves administration of
pharmaceutical compositions that include at least one compound of
Formula (I) or a pharmaceutically acceptable salt, active
metabolite, prodrug, or solvate thereof, in therapeutically
effective amounts to said subject.
[0211] In certain embodiments, the compositions containing the
compound(s) described herein are administered for prophylactic
and/or therapeutic treatments. In certain therapeutic applications,
the compositions are administered to a patient already suffering
from a disease or condition, in an amount sufficient to cure or at
least partially arrest at least one of the symptoms of the disease
or condition. Amounts effective for this use depend on the severity
and course of the disease or condition, previous therapy, the
patient's health status, weight, and response to the drugs, and the
judgment of the treating physician. Therapeutically effective
amounts are optionally determined by methods including, but not
limited to, a dose escalation clinical trial.
[0212] In prophylactic applications, compositions containing the
compounds described herein are administered to a patient
susceptible to or otherwise at risk of a particular disease,
disorder or condition.
[0213] In certain embodiments, the dose of drug being administered
may be temporarily reduced or temporarily suspended for a certain
length of time (i.e., a "drug holiday").
[0214] Doses employed for adult human treatment are typically in
the range of 0.01 mg-5000 mg per day or from about 1 mg to about
1000 mg per day. In one embodiment, the desired dose is
conveniently presented in a single dose or in divided doses.
Patient Selection
[0215] In any of the aforementioned aspects involving the
prevention or treatment of LPA-mediated diseases or conditions are
further embodiments comprising identifying patients by screening
for LPA receptor gene SNPs. Patients can be further selected based
on increased LPA receptor expression in the tissue of interest. LPA
receptor expression are determined by methods including, but not
limited to, northern blotting, western blotting, quantitative PCR
(qPCR), flow cytometry, autoradiography (using a small molecule
radioligand or PET ligand). In some embodiments, patients are
selected based on the concentration of serum or tissue LPA measured
by mass spectrometry. In some embodiments, patients are selected
based on a combination of the above markers (increased LPA
concentrations and increased LPA receptor expression).
Combination Treatments
[0216] In certain instances, it is appropriate to administer at
least one compound of Formula (I) in combination with another
therapeutic agent.
[0217] In one specific embodiment, a compound of Formula (I) is
co-administered with a second therapeutic agent, wherein the
compound of Formula (I) and the second therapeutic agent modulate
different aspects of the disease, disorder or condition being
treated, thereby providing a greater overall benefit than
administration of either therapeutic agent alone.
[0218] For combination therapies described herein, dosages of the
co-administered compounds vary depending on the type of co-drug(s)
employed, on the specific drug(s) employed, on the disease or
condition being treated and so forth. In additional embodiments,
when co-administered with one or more other therapeutic agents, the
compound provided herein is administered either simultaneously with
the one or more other therapeutic agents, or sequentially.
[0219] If administration is simultaneous, the multiple therapeutic
agents are, by way of example only, provided in a single, unified
form, or in multiple forms.
[0220] In another embodiment described herein, methods for
treatment of proliferative disorders, including cancer, comprises
administration to a mammal a compound of Formula (I) in combination
with one or more anti-cancer agents and/or radiation therapy.
[0221] In one aspect, compounds of Formula (I) are to treat or
reduce fibrosis in a mammal. In one aspect, compounds of Formula
(I) are administered in combination with one or more
immunosuppresants. In some embodiments, a compound of Formula (I)
is adminsitered with corticosteroids.
[0222] In yet another embodiment described herein, methods for
treating LPA-dependent or LPA-mediated conditions or diseases, such
as the therapy of respiratory disorders (e.g., pulmonary fibrosis,
asthma, COPD, rhinitis), comprises administration to a patient
compounds, pharmaceutical compositions, or medicaments described
herein in combination with at least one agent used in the treatment
of respiratory conditions.
[0223] In some embodiments, compounds of Formula (I) are
administered to a patient in combination with anti-inflammatory
agents.
[0224] In one embodiment, compounds of Formula (I) are administered
to a patient in combination with inhaled corticosteroids.
EXAMPLES
[0225] These examples are provided for illustrative purposes only
and not to limit the scope of the claims provided herein.
Synthesis of Compounds
Example 1
Synthesis of (R)-2'-chloro-alpha-methylbenzyl Alcohol
[0226] Using the procedure of Meier et at (Tetrahedron, 1996, 52,
589; Method 3), 2'-chloroacetophenone (Aldrich) was reduced to give
(R)-2'-chloro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLC
analysis of the acetate derivative (made by reacting the benzyl
alcohol with acetyl chloride and triethylamine in methylene
chloride) using Chiralcel OD eluted with 99:1 Hexane:ethanol. R
isomer retention time 4.3 minutes).
Example 2
Synthesis of (S)-2'-chloro-alpha-methylbenzyl Alcohol
[0227] Using the procedure of Meier et at (Tetrahedron, 1996, 52,
589; Method 3), 2'-chloroacetophenone (Aldrich) was reduced to give
(S)-2'-chloro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLC
analysis of the acetate derivative (made by reacting the benzyl
alcohol with acetyl chloride and triethylamine in methylene
chloride) using Chiralcel OD eluted with 99:1 Hexane:ethanol. S
isomer retention time 5.3 minutes)
Example 3
Synthesis of (R)-2'-fluoro-alpha-methylbenzyl Alcohol
[0228] Using the procedure of Meier et at (Tetrahedron, 1996, 52,
589; Method 3), 2'-fluoroacetophenone (Aldrich) was reduced to give
(R)-2'-fluoro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLC
analysis of the acetate derivative (made by reacting the benzyl
alcohol with acetyl chloride and triethylamine in methylene
chloride) using Chiralcel OD eluted with 99.8:0.2 Hexane:ethanol. R
isomer retention time 5.9 minutes).
Example 4
Synthesis of (S)-2'-fluoro-alpha-methylbenzyl Alcohol
[0229] Using the procedure of Meier et at (Tetrahedron, 1996, 52,
589; Method 3), 2'-fluoroacetophenone (Aldrich) was reduced to give
(S)-2'-fluoro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLC
analysis of the acetate derivative (made by reacting the benzyl
alcohol with acetyl chloride and triethylamine in methylene
chloride) using Chiralcel OD eluted with 99.8:0.2 Hexane:Ethanol. S
isomer retention time 6.7 minutes).
Example 5
Step 1: Synthesis of (3-Bromo-4-methoxy-phenyl)-acetic Acid Ethyl
Ester
[0230] To 3-bromo-4-methoxyphenylacetic acid (24 g, 97.9 mmol) in
ethanol (240 mL) was added thionyl chloride (7.8 mL, 107.7 mmol),
and the reaction was stirred at room temperature for 1 hour. Once
no starting material was seen by analytical LCMS, the mixture was
basified with saturated aqueous NaHCO.sub.3 and extracted with
dichloromethane. The combined organic layers were dried over
MgSO.sub.4, filtered, and concentrated to give the desired
product.
Step 2: Synthesis of
[4-Methoxy-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceti-
c acid ethyl ester
[0231] (3-Bromo-4-methoxy-phenyl)-acetic acid ethyl ester (27.4 g,
100.3 mmol), bis(pinacolato)diboron (25.47 g, 100.3 mmol), and
potassium acetate (24.6 g, 250.8 mmol) were combined in 1,4-dioxane
(250 mL) under N.sub.2. The solution was purged with N.sub.2, and
then (1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II)
(4.10 g, 5.02 mmol) was added and the reaction was heated to
110.degree. C. overnight. The mixture was filtered through Celite
and partitioned between EtOAc and brine. The aqueous layer was
separated and extracted twice with EtOAc, and the combined organic
layers were dried and concentrated. The residue was purified by
silica gel chromatography (20-60% EtOAc in hexanes) to give the
title compound.
Example 6
Synthesis of 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic
Acid
Step 1: 3-Methylamino-but-2-enoic Acid Methyl Ester
[0232] To a solution of methyl acetoacetate (29.4 g, 253 mmol) in
methanol (30 mL) was added methylamine (33 wt % in EtOH; 48 mL, 385
mmol) dropwise at room temperature. The reaction was stirred for 1
hour, and then concentrated and dried to give the title compound as
a white crystalline solid.
Step 2: 2-(4-Bromo-benzoyl)-3-oxo-butyric Acid Methyl Ester
[0233] To 3-methylamino-but-2-enoic acid methyl ester (5.0 g, 39.1
mmol) in THF (70 mL) was added pyridine (3.7 mL, 47 mmol) dropwise.
The mixture was cooled to 0.degree. C., and 4-bromobenzoyl chloride
(8.55 g, 39.1 mmol) in THF (30 mL) was added dropwise. The reaction
was stirred at room temperature overnight, and then water was
added. The mixture was extracted with EtOAc, and the combined
organic layers were washed with water, dried, filtered, and
concentrated to give the title compound.
Step 3: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic Acid
Methyl Ester
[0234] To a mixture of 2-(4-bromo-benzoyl)-3-oxo-butyric acid
methyl ester (11 g, 39 mmol) in acetic acid (50 mL) was added
hydroxylamine hydrochloride (2.66 g, 39 mmol), and the reaction was
stirred at 115.degree. C. for 1 hours. After cooling, saturated
aqueous NaHCO.sub.3 was added to the mixture to adjust to pH 8. The
solution was extracted with EtOAc, and the combined organic layers
were washed with brine, dried, filtered, and concentrated to give
the title compound.
Step 4: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic Acid
[0235] Lithium hydroxide (2 g, 48 mmol) was added to a solution of
5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid methyl
ester (39 mmol) in methanol (50 mL) and water (10 mL), and the
reaction was stirred at 60.degree. C. for 1 hour. Acidic work-up
gave the title compound.
Example 7
Synthesis of
[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
Acid Ethyl Ester and 4-((Ethoxycarbonyl)methyl)phenylboronic
Acid
Step 1:
[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
Acid Ethyl Ester
[0236] Ethyl 4-bromophenylacetate (12 g, 49 mmol),
bis(pinacolato)diboron (12.4 g, 59 mmol),
1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (2 g,
2.4 mmol), and potassium acetate (9.6 g, 98 mmol) were combined in
1,4-dioxane (100 mL), and the reaction was heated to 80.degree. C.
for 6 hours. The mixture was worked-up to give the title
compound.
Step 2: 4-((Ethoxycarbonyl)methyl)phenylboronic Acid
[0237]
[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl (0.460 g, 1.58 mmol), sodium periodate (0.746 g, 3.49
mmol), and ammonium acetate (0.309 g, 4.01 mmol) were combined in
1:1 acetone:water and stirred overnight at room temperature.
Aqueous work-up provided the title compound.
Example 8
Synthesis of
[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
Acid Methyl Ester and 3-((Methoxycarbonyl)methyl)phenylboronic
Acid
Step 1:
[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
Acid Methyl Ester
[0238] Prepared according to the procedure described in Example 7,
Step 1 using methyl 2-(3-bromophenyl)acetate and
bis(pinacolato)diboron.
Step 2: 3-((Methoxycarbonyl)methyl)phenylboronic Acid
[0239] Prepared according to the procedure described in Example 7,
Step 2 using
[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid methyl ester.
Example 9
Synthesis of
(4'-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 1)
Step 1:
5-(4'-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-car-
boxylic acid
[0240] 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (2.0
g, 7.07 mmol),
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester (2.46 g, 8.5 mmol),
tetrakis(triphenylphosphine)palladium(0) (0.80 g, 0.70 mmol), and
sodium bicarbonate (4.3 g, 52 mmol) were combined in 1,4-dioxane
(50 mL) and water (10 mL), and the reaction was stirred overnight
at 80.degree. C. to give the title compound.
Step 2:
(4'-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-
-isoxazol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester
[0241]
5-(4'-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carb-
oxylic acid (0.150 g, 0.41 mmol),
alpha-methyl-2-trifluoromethylbenzyl alcohol (0.094 g, 0.49 mmol),
diphenylphosphoryl azide (0.11 mL, 0.49 mmol), and triethylamine
(0.11 mL, 0.49 mmol) were combined in toluene (3 mL) and stirred at
80.degree. C. for 2 hours. After cooling, the mixture was worked-up
to give the title compound.
Step 3:
(4'-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-
-isoxazol-5-yl}-biphenyl-4-yl)-acetic Acid
[0242]
(4'-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]--
isoxazol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester (0.41 mmol)
was treated with aqueous lithium hydroxide in methanol at
60.degree. C. for 30 minutes. The mixture was purified by
preparative HPLC to give the title compound.
Example 10
Synthesis of
(4'-{3-Methyl-4-[1-(3-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic Acid (Compound 2)
[0243] Following the procedure described in Example 9, Step 2,
5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and alpha-methyl-3-trifluoromethylbenzyl alcohol were reacted
to provide
(4'-{3-methyl-4-[1-(3-trifluoromethyl-phenyl)-ethoxycarbonylamino-
]-isoxazol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 9, Step 3.
Example 11
Synthesis of
(4'-{4-[1-(2,4-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic Acid (Compound 3)
[0244] Following the procedure described in Example 9, Step 2,
5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and 2,4-dichloro-alpha-methylbenzyl alcohol were reacted to
provide
(4'-{4-[1-(2,4-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 9, Step 3.
Example 12
Synthesis of
(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic Acid (Compound 4)
[0245] Following the procedure described in Example 9, Step 2,
5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and 2-fluoro-alpha-methylbenzyl alcohol were reacted to
provide
(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to the
acid as described in Example 9, Step 3.
Example 13
Synthesis of
(4'-{4-[1-(3-Bromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-4-yl)-acetic Acid (Compound 5)
[0246] Following the procedure described in Example 9, Step 2,
5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and 3-bromo-alpha-methylbenzyl alcohol were reacted to provide
(4'-{4-[1-(3-bromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to the
acid as described in Example 9, Step 3.
Example 14
Synthesis of
(4'-{4-[1-(2-Methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-
-biphenyl-4-yl)-acetic Acid (Compound 6)
[0247] Following the procedure described in Example 9, Step 2,
5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and 1-(2-methoxyphenyl)ethanol were reacted to provide
(4'-{4-[1-(2-Methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-
-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 9, Step 3.
Example 15
Synthesis of (3-Bromo-4-fluoro-phenyl)-acetic Acid Ethyl Ester
[0248] Step 1: 3-Bromo-4-fluorophenylacetic acid (2.35 g, 9.0 mmol)
was treated with concentrated sulfuric acid (2 mL) in ethanol (100
mL) at reflux for 2 hours. Purification by silica gel
chromatography provided the title compound.
Example 16
Synthesis of (5-Bromo-2-fluoro-phenyl)-acetic Acid Ethyl Ester
[0249] Step 1: Prepared according to the procedure described in
Example 15, Step 1 using the following starting material:
5-bromo-2-fluorophenylacetic acid.
Example 17
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
6-methoxy-biphenyl-3-yl)-acetic Acid (Compound 7)
[0250] Step 1: To
5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (5.764 g,
20.4 mmol) in toluene (200 mL) was added triethylamine (3.13 mL,
22.4 mmol) and diphenylphosphoryl azide (4.42 mL, 20.4 mmol), and
the mixture was stirred for 5 minutes at room temperature.
2-Chloro-alpha-methylbenzyl alcohol (3.2 g, 22.4 mol) was added,
and the reaction was stirred at 80.degree. C. for 2 hours. The
mixture was diluted with EtOAc (200 mL) and extracted with EtOAc.
The combined organic layers were washed twice with water and once
with brine, and the combined aqueous layers were back-extracted
with EtOAc. The combined organic layers were concentrated, and the
residue was purified by silica gel chromatography to give
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-chloro-phenyl)-ethyl ester.
[0251] Step 2: To
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-chloro-phenyl)-ethyl ester (0.100 g, 0.23 mmol) in DME (5 mL)
was added
[4-methoxy-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phe-
nyl]-acetic acid ethyl ester (0.074 g, 0.23 mmol), followed by
potassium carbonate (0.079 g, 0.58 mmol) and water (3 mL). The
solution was purged with N.sub.2 for 10 minutes, and then
tetrakis(triphenylphosphine)palladium(0) (0.027 g, 0.02 mmol) was
added. The reaction was stirred at 100.degree. C. until no starting
material was seen by analytical LCMS and thin layer chromatography
(tic), and then cooled to room temperature and diluted with EtOAc.
The mixture was extracted with EtOAc, and the combined organic
layers were washed with water and brine. The combined aqueous
layers were back-extracted with EtOAc, and the combined organic
layers were dried and concentrated. The residue was purified by
silica gel chromatography to give
(4'-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
6-methoxy-biphenyl-3-yl)-acetic acid ethyl ester.
[0252] Step 3: To
(4'-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
6-methoxy-biphenyl-3-yl)-acetic acid ethyl ester (0.105 g, 0.20
mmol) in methanol (20 mL) was added 1N aqueous LiOH (5 mL, 5 mmol),
and the reaction was stirred for 2 hours. The mixture was diluted
with water and extracted with EtOAc. The combined organic layers
were dried and concentrated to give the title compound.
Example 18
Synthesis of
{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]isox-
azol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester
[0253] [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-chloro-phenyl)-ethyl ester (3.48 g, 8.0 mmol),
bis(pinacolato)diboron (2.24 g, 8.8 mmol), and potassium acetate
(2.88 g, 32.0 mmol) were combined in 1,4-dioxane (200 mL) under
N.sub.2. The solution was purged with N.sub.2 for 10 minutes, and
then (1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II)
(0.652 g, 0.8 mmol) was added and the reaction was heated to
80.degree. C. for 6 hours, and then cooled to room temperature and
stirred overnight. The mixture was diluted with water and extracted
with EtOAc. The combined organic layers were washed twice with
water and once with brine, and the combined aqueous layers were
back-extracted with EtOAc. The combined organic layers were
concentrated and the residue was purified by silica gel
chromatography (0-100% EtOAc in hexanes) to give the title
compound.
Example 19
Synthesis of
4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino}-3-methyl-isoxazol-5-yl]-b-
iphenyl-4-carboxylic acid (Compound 8)
[0254] Step 1:
{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester (0.100 g,
0.21 mmol), 4-bromobenzoic acid (0.042 g, 0.21 mmol), and potassium
carbonate (0.073 g, 0.53 mmol) were combined in DME (10 mL) and
water (5 mL). The solution was purged with N.sub.2 for 10 minutes,
and then
(1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (0.014
g, 0.02 mmol) was added. The reaction was stirred at 70.degree. C.
overnight, until no starting material was seen by analytical LCMS.
The mixture was cooled to room temperature and quenched with water.
The mixture was extracted with EtOAc, and the combined organic
layers were dried and concentrated. The residue was purified by
preparative HPLC to give the title compound.
Example 20
Synthesis of
4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-2-carboxylic acid (Compound 9)
[0255] Step 1:
{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester, methyl
2-bromobenzoate, and bis(triphenylphosphine)palladium(II)
dichloride were reacted as described in Example 17, Step 2 to
provide
4'-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-2-carboxylic acid methyl ester.
[0256] Step 2: To
4'-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-2-carboxylic acid methyl ester (0.21 mmol) in methanol was
added 1N aqueous LiOH (2 mL) and stirred overnight at room
temperature. The solution was stirred at 40.degree. C. for 3 days.
The mixture was purified by preparative HPLC to give the title
compound.
Example 21
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-2-yl)-acetic Acid (Compound 10)
[0257] Step 1: Prepared according to the procedure described in
Example 19, Step 1 using
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester and
2-bromophenylacetic acid.
Example 22
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic Acid (Compound 11)
[0258] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester, ethyl
4-bromophenylacetate, and
(1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) were
reacted to provide (4'-{-4-[1-(2-chloro-phenyl)-ethoxyc
arbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-acetic acid
ethyl ester, which was hydrolyzed to the acid as described in
Example 17, Step 3.
Example 23
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-3-yl)-acetic acid (Compound 12)
[0259] 3-Bromophenylacetic acid (0.3 .mu.mol) was treated with
concentrated sulfuric acid in ethanol at reflux for 2 hours to give
(3-bromo-phenyl)-acetic acid ethyl ester.
[0260] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
(3-bromo-phenyl)-acetic acid ethyl ester, and
(1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) were
reacted to provide (4'-{-4-[1-(2-Chloro-phenyl)-ethoxyc
arbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-3-yl)-acetic acid
ethyl ester, which was hydrolyzed to the acid as described in
Example 17, Step 3.
Example 24
Synthesis of
3-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid (Compound 13)
[0261] 3-(4-Bromo-phenyl)-propionic acid ethyl ester was prepared
according to the procedure described in Example 15, Step 1 using
3-(4-bromophenyl)propionic acid.
[0262] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
3-(4-bromo-phenyl)-propionic acid ethyl ester, and
bis(triphenylphosphine)palladium(II) dichloride were reacted to
provide 3-(4'-{4-[1-(2-chloro-phenyl)-ethoxyc arb
onylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionic acid
ethyl ester, which was hydrolyzed to the acid as described in
Example 17, Step 3.
Example 25
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
6-fluoro-biphenyl-3-yl)-acetic acid (Compound 14)
[0263] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
(3-bromo-4-fluoro-phenyl)-acetic acid ethyl ester, and
(1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) were
reacted to provide (4'-{4-[1-(2-chloro-phenyl)-ethoxyc
arbonylamino]-3-methyl-isoxazol-5-yl}-6-fluoro-biphenyl-3-yl)-acetic
acid ethyl ester, which was hydrolyzed to the acid as described in
Example 17, Step 3.
Example 26
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
4-fluoro-biphenyl-3-yl)-acetic Acid (Compound 15)
[0264] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
(5-bromo-2-fluoro-phenyl)-acetic acid ethyl ester, and
(1,1'-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) were
reacted to provide (4'-{4-[1-(2-chloro-phenyl)-ethoxyc
arbonylamino]-3-methyl-isoxazol-5-yl}-4-fluoro-biphenyl-3-yl)-acetic
acid ethyl ester, which was hydrolyzed to the acid as described in
Example 17, Step 3.
Example 27
Synthesis of
(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}--
biphenyl-4-yl)-acetic Acid Methyl Ester (Compound 16)
[0265] To
(4'-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid (0.050 g, 0.1 mmol) in benzene
(17 mL) and methanol (2 mL) was added (trimethylsilyl)diazomethane
(2M in hexanes; 0.1 mL, 0.2 mmol), and the reaction was stirred at
room temperature for 5 minutes. Additional
(trimethylsilyl)diazomethane (2M in hexanes; 0.25 mL, 0.5 mmol) was
added, and the reaction was stirred until no starting material was
seen by analytical LCMS. The mixture was quenched with water and
extracted with EtOAc. The combined organic layers were washed twice
with water and once with brine, and then dried and concentrated.
The residue was purified by silica gel chromatography (0-80% EtOAc
in hexanes) to give the title compound.
Example 28
Synthesis of
2-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino}-3-methyl-isoxazol-5-yl-
]-biphenyl-4-yl)-propionic Acid Ethyl Ester (Compound 17)
[0266] Step 1: To ethyl 4-bromophenylacetate (2.137 g, 8.8 mmol) in
THF (20 mL) at -78.degree. C. was added lithium
bis(trimethylsilyl)amide (1M in hexane; 11.4 mL, 11.4 mmol), and
the solution was stirred for 1 hour at -78.degree. C. Iodomethane
(0.71 mL, 11.4 mmol) in THF (5 mL) was added, and the reaction was
warmed to room temperature. The mixture was quenched with water and
concentrated. The residue was dissolved in EtOAc and washed three
times with water and once with brine. The organic layer was dried
and concentrated, and the crude material was purified by silica gel
chromatography (0-60% EtOAc in hexanes) to give
2-(4-bromo-phenyl)-propionic acid ethyl ester.
[0267] Step 2: Following the procedure described in Example 17,
Step 2 the title compound was prepared using
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
2-(4-bromo-phenyl)-propionic acid ethyl ester, and
bis(triphenylphosphine)palladium(II) dichloride; further
purification by preparative HPLC was required.
Example 29
Synthesis of
2-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic Acid (Compound 18)
[0268] Prepared according to the procedure described in Example 17,
Step 3 using
2-(4'-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazo-
l-5-yl}-biphenyl-4-yl)-propionic acid ethyl ester.
Example 30
Synthesis of
2-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 19)
[0269] To sodium hydride (0.859 g, 35.8 mmol) and iodomethane (2.23
mL, 35.8 mmol) in dimethylformamide (80 mL) at 0.degree. C. was
added ethyl 4-bromophenylacetate (2.17 g, 9.0 mmol) dropwise, and
the reaction was warmed to room temperature and stirred for 4
hours. The mixture was quenched with water and 1N aqueous HCl (20
mL), and then extracted with 1:1 EtOAc:hexanes. The combined
organic layers were washed five times with water and once with
brine, and the combined aqueous layers were back-extracted with
EtOAc. The combined organic layers were dried and concentrated, and
the residue was purified by silica gel chromatography (0-20% EtOAc
in hexanes). Further purification by preparative HPLC provided
2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester.
[0270] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester, and
bis(triphenylphosphine)palladium(II) dichloride were reacted to
provide 2-(4'-{4-[1-(2-chloro-phenyl)-ethoxycarb
onylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionic
acid ethyl ester; further purification by preparative HPLC was
required. The ester was hydrolyzed to the acid as described in
Example 17, Step 3; purification by preparative HPLC was
utilized.
Example 31
Synthesis of
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid (Compound 20)
[0271] Step 1: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic
acid (4 g, 14.2 mmol) was dissolved in toluene (150 mL).
Triethylamine (2.187 mL, 15.6 mmol) was added, followed by
diphenylphosphoryl azide (3.372 mL, 15.6 mmol), and the mixture was
stirred for 10 minutes. 2-Fluoro-alpha-methylbenzyl alcohol (2 g,
15.6 mmol) was added, and the reaction was stirred at 80.degree. C.
overnight. The mixture was cooled to room temperature and
concentrated, and the residue was partitioned between with EtOAc
and water. The organic layer was washed 4 times with water and once
with brine, and then dried, filtered, and concentrated, and the
residue was purified by silica gel chromatography (dry load; 0-100%
EtOAc in hexanes) to give
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-fluoro-phenyl)-ethyl ester.
Step 2:
{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phen-
yl]-isoxazol-4-yl}-carbamic acid 1-(2-fluoro-phenyl)-ethyl
Ester
[0272] Prepared according to the procedure described in Example 18
using [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-fluoro-phenyl)-ethyl ester and bis(pinacolato)diboron.
Step 3:
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-propionic Acid Ethyl Ester
[0273] Prepared according to the procedure described in Example 17,
Step 2 using
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pheny-
l]-isoxazol-4-yl}-carbamic acid 1-(2-fluoro-phenyl)-ethyl ester,
2-(4-bromo-phenyl)-propionic acid ethyl ester, and
bis(triphenylphosphine)palladium(II) dichloride; purification by
preparative HPLC, followed by silica gel chromatography, was
utilized.
Step 4:
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino}-3-methyl-isoxaz-
ol-5-yl]-biphenyl-4-yl)-propionic acid
[0274] Prepared as described in Example 17, Step 3 using
2-(4'-{4-[1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-propionic acid ethyl ester.
Example 32
Synthesis of
4-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-butyric acid (Compound 21)
[0275] 4-(4-Bromo-phenyl)-butyric acid ethyl ester was prepared
according to the procedure described in Example 23 using
4-(4-bromophenyl)butanoic acid.
[0276] Following the procedure described in Example 17, Step 2,
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-chloro-phenyl)-ethyl ester,
4-(4-bromo-phenyl)-butyric acid ethyl ester, and
bis(triphenylphosphine)palladium(II) dichloride were reacted to
provide
4-(4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-butyric acid ethyl ester, which was hydrolyzed to
the acid as described in Example 17, Step 3.
Example 33
Synthesis of
4'-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-b-
iphenyl-3-carboxylic acid (Compound 22)
[0277] Step 1: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic
acid 1-(2-chloro-phenyl)-ethyl ester (0.200 g, 0.46 mmol),
3-carboxyphenylboronic acid (0.092 g, 0.55 mmol), and potassium
carbonate (0.190 g, 1.37 mmol) were combined in 2:1 DME:water (5
mL), and the solution was purged with N.sub.2 for 10 minutes.
Tetrakis(triphenylphosphine)palladium(0) (0.053 g, 0.05 mmol) was
added, and the reaction was purged with N.sub.2 for an additional
10 minutes, and then sealed and stirred at 80.degree. C. overnight.
The mixture was partitioned between EtOAc and water, and the
aqueous layer was extracted with EtOAc. The combined organic layers
were dried over MgSO.sub.4, filtered, and concentrated, and the
residue was purified by preparative HPLC to give the title
compound.
Example 34
Synthesis of
(4'-{4-[1-(4-Chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic Acid (Compound 23)
[0278] Step 1:
5-(4'-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and 1-(4-chloro-2-fluorophenyl)ethanol were reacted as
described in Example 31, Step 1 to provide
(4'-{4-[1-(4-Chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester.
[0279] Step 2: Excess lithium hydroxide was added to a solution of
(4'-{4-[1-(4-chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester (0.100 g, 0.19
mmol) in methanol and water, and the reaction was stirred at room
temperature overnight. The mixture was acidified with 1N aqueous
HCl and extracted with EtOAc, and the combined organic layers were
dried, filtered, and concentrated. The residue was purified by
preparative HPLC to give the title compound.
Example 35
Synthesis of
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic Acid (Compound 24)
[0280] Step 1: 2'-Fluoroacetophenone (25 g, 180 mmol) and
(S)-(-)-2-methyl-CBS-- oxazaborolidine (6.02 g, 22 mmol) were
combined in THF (200 mL). Borane methyl sulfide complex (2M in THF;
59.4 mL, 119 mmol) was added over 20 minutes, and the reaction was
stirred at room temperature for 30 minutes. Methanol was added, and
the mixture was worked-up with dichloromethane and water. The
organic layer was concentrated to give
(R)-1-(2-fluoro-phenyl)-ethanol in 90.6% e.e.
[0281] Step 2:
5-(4'-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and (R)-1-(2-fluoro-phenyl)-ethanol were reacted as described
in Example 31, Step 1 to provide
(4'-{4-[(R)-1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester.
[0282] Step 3: The title compound was prepared as described in
Example 34, Step 2 using
(4'-{4-[(R)-1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester.
Example 36
Synthesis of
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-2'-methyl-biphenyl-4-yl)-acetic Acid (Compound 25)
Step 1: 4-Bromo-3-methyl-benzoyl Chloride
[0283] To 4-bromo-3-methylbenzoic acid (8.6 g, 40.0 mmol) in
CH.sub.2Cl.sub.2 was added DMF (0.4 mL), followed by oxalyl
chloride (3.7 mL, 42 mmol) slowly over 10 minutes by syringe. The
reaction was stirred for 1 hour at room temperature, and then
concentrated to give the title compound.
Step 2: 2-(4-Bromo-3-methyl-benzoyl)-3-[(E)-methylimino]-butyric
Acid Methyl Ester
[0284] To 4-bromo-3-methyl-benzoyl chloride (40.0 mmoml) in
tertahydrofuran (300 mL) wad added 3-methylamino-but-2-enoic acid
methyl ester (5.12 g, 40.0 mmol), followed by pyridine (3.2 mL,
40.0 mmol), and the reaction was stirred overnight at room
temperature. Solid pyridine hydrochloride coated the flask, so the
mixture was decanted, and the solid was washed three times with
EtOAc. The combined solutions were concentrated and diluted with
EtOAc (500 mL), and the mixture was washed three times with water.
The aqueous layer was separated and back-extracted with EtOAc, and
the combined organic layers were dried over MgSO.sub.4, filtered,
and concentrated to give the title compound.
Step 3: 5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic
Acid Methyl Ester
[0285] 2-(4-Bromo-3-methyl-benzoyl)-3-[(E)-methylimino]-butyric
acid methyl ester (40.0 mmol) and hydroxylamine hydrochloride (2.9
g, 41 mmol) were combined in acetic acid (100 mL) and stirred at
60.degree. C. overnight. The mixture was concentrated, and the
residue was partitioned between EtOAc and water. Saturated aqueous
NaHCO.sub.3 was added to neutralize residual acetic acid, and the
organic layer was separated and washed twice with water. The
combined aqueous layers were back-extracted with EtOAc, and the
combined organic layers were dried, filtered, and concentrated. The
crude material was purified by silica gel chromatography to give
the title compound.
Step 4: 5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic
Acid
[0286] 5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic
acid methyl ester (7.26 g, 23.4 mmol) was hydrolyzed with 1N
aqueous LiOH. Acidic work-up provided the title compound.
Step 5:
[5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazol-4-yl]carbamic acid
(R)-1-(2-fluoro-phenyl)-ethyl Ester
[0287] 5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic
acid (1.78 g, 6.0 mmol) was dissolved in toluene (50 mL).
Triethylamine (0.92 mL, 6.6 mmol) was added, followed by
diphenylphosphoryl azide (1.43 mL, 6.6 mmol), and the mixture was
stirred for 10 minutes. (R)-1-(2-Fluoro-phenyl)-ethanol (0.924 g,
6.6 mmol) was added, and the reaction was stirred at 85.degree. C.
for 3 hours, and then overnight at room temperature. The mixture
was concentrated, and the residue was diluted with dichloromethane
and purified by silica gel chromatography to give the title
compound.
Step 6:
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isox-
azol-5-yl}-2'-methyl-biphenyl-4-yl)-acetic Acid Ethyl Ester
[0288]
[5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-fluoro-phenyl)-ethyl ester (0.350 g, 0.80 mmol),
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester (0.292 g, 0.96 mmol), and sodium bicarbonate
(0.235 g, 2.8 mmol) were combined in 2:1 DME:water (11 mL), and the
solution was purged with N.sub.2 for 5 minutes.
Bis(triphenylphosphine)palladium(II) dichloride (0.030 g, 0.04
mmol) was added, and the reaction was stirred at 80.degree. C. for
2 hour. After work-up, the crude material was purified by silica
gel chromatography to give the title compound.
Step 7:
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isox-
azol-5-yl}-2'-methyl-biphenyl-4-yl)-acetic Acid
[0289]
(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxa-
zol-5-yl}-2'-methyl-biphenyl-4-yl)-acetic acid ethyl ester (0.282
g, 0.53 mmol) was hydrolyzed with lithium hydroxide in methanol and
water, and the crude material was purified by preparative HPLC to
give the title compound.
Example 37
Synthesis of
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl-
}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 26)
Step 1:
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-2-methyl-propionic Acid Ethyl Ester
[0290] Prepared as described in Example 36, Step 6 using
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid 1-(2-fluoro-phenyl)-ethyl ester and
2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester;
purification by preparative HPLC was utilized.
Step 2:
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-2-methyl-propionic Acid
[0291]
2-(4'-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazo-
l-5-yl}-biphenyl-4-yl)-2-methyl-propionic acid ethyl ester (0.51
mmol) in Methanol (50 mL) was treated with lithium hydroxide
(excess) as well as 1N aqueous LiOH (excess, and the reaction was
stirred at room temperature. Analytical LCMS indicated that
starting material was still present, so the reaction was stirred at
80.degree. C. overnight. The mixture was neutralized with 1N HCl
(20 mL) and extracted with EtOAc. The combined organic layers were
washed with water, dried, and concentrated, and the residue was
purified by preparative HPLC to give the title compound.
Example 38
Synthesis of
(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 27)
Step 1: (R)-1-(2-Chloro-phenyl)-ethanol
[0292] 2'-Chloroacetophenone (8.4 mL, 65 mmol) and
(S)-(-)-2-methyl-CBS-- oxazaborolidine (0.90 g, 0.32 mmol) were
combined in THF (75 mL). Borane methyl sulfide complex (2M in THF;
21.5 mL, 43 mmol) was added over 20 minutes, and the reaction was
stirred at room temperature for 30 minutes. MeOH was added, and the
mixture was worked-up with CH.sub.2Cl.sub.2 and H.sub.2O. The
organic layer was concentrated to give the title compound.
Step 2: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl Ester
[0293] 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (10
g, 35 mmol), (R)-1-(2-chloro-phenyl)-ethanol (6.6 g, 42 mmol),
triethylamine (10 mL, 70 mmol), and diphenylphosphoryl azide (11.5
g, 42 mmol) were combined in toluene (100 mL) and stirred at
90.degree. C. for 1 hour. The mixture was concentrated, and the
residue was purified by silica gel chromatography (0-30% EtOAc in
hexanes). The isolated product was recrystallized in 5:1
hexanes:acetone to give the title compound in >99% e.e (by
chiral HPLC. Chiracel OD 98.4% hexanes/1.6% Ethanol. Major isomer
27.9 min minor isomer 32.7 min).
Step 3:
[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
Acid Ethyl Ester
[0294] Ethyl 4-bromophenylacetate (9.72 g, 40 mmol),
bis(pinacolato)diboron (12.2 g, 48 mmol), and potassium acetate
(18.8 g, 1690 mmol) were combined in 1,4-dioxane, and the mixture
was purged with N.sub.2 for 10 minutes.
1,1'-Bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (3.26
g, 4.0 mmol) was added, and the reaction was heated to 80.degree.
C. for 24 hours, and at room temperature for 2 days. The mixture
was diluted with H.sub.2O and extracted with EtOAc. The combined
organic layers were washed with 4 times with H.sub.2O, once with
brine, and then dried, filtered, and concentrated. The residue was
dissolved in CH.sub.2Cl.sub.2 and filtered through Celite to remove
solids. The filtrate was concentrated, and the crude material was
purified by silica gel chromatography (0-30% EtOAc in hexanes) to
give the title compound.
Step 4:
(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isox-
azol-5-yl}-biphenyl-4-yl)-acetic Acid Ethyl Ester
[0295] [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester (3.0 g, 6.9 mmol),
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester (2.2 g, 7.6 mmol),
dichloro-bis(triphenylphosphine)palladium(II) (0.25 g, 0.35 mmol),
and sodium bicarbonate (1.7 g, 20.7 mmol) were combined in DME (30
mL) and H.sub.2O (10 mL), and the reaction was stirred at
80.degree. C. for 2 hours. After aqueous work-up, the crude
material was purified by silica gel chromatography (0-40% EtOAc in
hexanes) to give the title compound.
Step 5:
(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isox-
azol-5-yl}-biphenyl-4-yl)-acetic acid
[0296]
4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxaz-
ol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester (3 g, 5.9 mmol) in
MeOH and H.sub.2O was treated with lithium hydroxide (1 g, 23.8
mmol), and the reaction was stirred at 60.degree. C. Acidic work-up
provided the title compound.
Example 39
Synthesis of
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-2-methyl-propionic acid (Compound 28)
Step 1:
2-Methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pheny-
l]-propionic Acid Ethyl Ester
[0297] Prepared as described in Example 18 using
2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester and
bis(pinacolato)diboron.
Step 2:
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-is-
oxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionic acid ethyl ester
[0298] Prepared as described in Example 36, Step 6 using
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and
2-methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-prop-
ionic acid ethyl ester.
Step 3:
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-is-
oxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionic Acid
[0299] Prepared as described in Example 17, Step 3 using
2-(4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-2-methyl-propionic acid ethyl ester.
Example 40
Synthesis of
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-2-methyl-propionic Acid (Compound 29)
[0300] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-fluoro-phenyl)-ethyl ester and
2-methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-prop-
ionic acid ethyl ester were reacted to provide
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-2-methyl-propionic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 17, Step 3.
Example 41
Synthesis of
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic Acid (Compound 30)
[0301]
2-[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propion-
ic acid ethyl ester was prepared according to the procedure
described in Example 18 using 2-(4-bromo-phenyl)-propionic acid
ethyl ester and bis(pinacolato)diboron.
[0302] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-fluoro-phenyl)-ethyl ester and
2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionic
acid ethyl ester were reacted to provide
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 17, Step 3.
Example 42
Synthesis of
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic Acid (Compound 31)
[0303] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and
2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionic
acid ethyl ester were reacted to provide
2-(4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 17, Step 3.
Example 43
Synthesis of
[5-(4'-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
Acid (R)-1-(2-chloro-phenyl)-ethyl Ester (Compound 61)
[0304] Prepared as described in Example 36, Step 6 using
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and 4-cyanomethylphenylboronic
acid.
Example 44
Synthesis of
[5-(4'-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
Acid (R)-1-(2-fluoro-phenyl)-ethyl Ester (Compound 62)
[0305] Prepared as described in Example 36, Step 6 using
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-fluoro-phenyl)-ethyl ester and 4-cyanomethylphenylboronic
acid.
Example 45
Synthesis of
{3-Methyl-[5-(4'-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-c-
arbamic acid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 63)
[0306] To a solution of
[5-(4'-cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-fluoro-phenyl)-ethyl ester (0.200 g, 0.44 mmol) in
toluene (5 mL) was added dibutyltin oxide (0.011 g, 0.04 mmol),
followed by azidotrimethylsilane (0.07 mL, 0.53 mmol), and the
reaction was stirred at 100.degree. C. for 5 hours, and then
stirred at room temperature overnight. The mixture was
concentrated, and the residue was dissolved in EtOAc and washed
with water and brine. The organic layer was dried and concentrated,
and the crude material was purified by preparative HPLC to give the
title compound.
Example 46
Synthesis of
{3-Methyl-5-[4'-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-ca-
rbamic acid (R)-1-(2-chloro-phenyl)-ethyl ester (Compound 64)
[0307] Prepared as described in Example 45 using
[5-(4'-cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-chloro-phenyl)-ethyl ester and
azidotrimethylsilane.
Example 47
Synthesis of
{4'-[3-Methyl-4-(R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-
-4-yl}-acetic Acid (Compound 37)
Step 1: (R)-1-Phenyl-ethanol
[0308] To acetophenone (19.47 mL, 166.6 mmol) in THF (100 mL) was
added (S)-(-)-2-methyl-CBS-oxazaborolidine (4.62 g, 16.6 mmol), and
the reaction was cooled to 0.degree. C. Borane methyl sulfide
complex (2M in THF; 50 mL, 100 mmol) was added over 15 minutes, and
the reaction was stirred at room temperature. Aqueous work-up gave
the title compound.
Step 2: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-phenyl-ethyl Ester
[0309] 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (25
g, 88.7 mmol) in toluene (500 mL) was added triethylamine (18.5 mL,
133 mmol), followed by diphenylphosphoryl azide (22.1 mL, 101.9
mmol). (R)-(+)-1-Phenylethyl alcohol (11.9 mL, 97.5 mmol) was
added, and the reaction was stirred at 75.degree. C. for 2 hours.
The mixture was partitioned between EtOAc and H.sub.2O and filtered
through Celite. The aqueous layer was extracted with EtOAc, and the
combined organic layers were dried over MgSO.sub.4, filtered, and
concentrated to give the title compound.
Step 3:
{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl}--
biphenyl-4-yl]-acetic Acid Ethyl Ester
[0310] [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-phenyl-ethyl ester (39 g, 97.2 mmol),
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester (31 g, 107 mmol), and sodium bicarbonate (32.6 g,
389 mmol) were combined in 3:1 DME:H.sub.2O (500 mL), and the
mixture was purged with N.sub.2 for 15 minutes.
(1,1'-Bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (2.13
g, 2.91 mmol) was added, and the reaction was purged with N.sub.2
for an additional 10 minutes and then stirred at 90.degree. C.
overnight. The mixture was partitioned between EtOAc and H.sub.2O,
and the aqueous layer was extracted with EtOAc. The combined
organic layers were washed with H.sub.2O, dried over MgSO.sub.4,
filtered, and concentrated, and the residue was purified by silica
gel chromatography (EtOAc/hexane gradient) to give the title
compound.
Step 4:
{4'-[3-Methyl-4-(R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-b-
iphenyl-4-yl}-acetic Acid
[0311] To a suspension of
{4'-[3-methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-4-yl}-acetic acid ethyl ester (24 g, 49 mmol) in 3:1
MeOH:H.sub.2O (300 mL) was added lithium hydroxide (8.3 g, 198
mmol), and the reaction was stirred at room temperature overnight.
The mixture was acidified and partitioned between EtOAc and
H.sub.2O. The aqueous layer was extracted with EtOAc, and the
combined organic layers were washed with H.sub.2O, dried over
MgSO.sub.4, filtered, and concentrated to give the title compound.
Mass spec. data (M+H)=457.
Example 48
Synthesis of
{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-3-yl}-acetic Acid (Compound 43)
[0312] (3-Bromo-phenyl)-acetic acid ethyl ester and
bis(pinacolato)diboron were reacted as described in Example 18 to
provide
[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester.
[0313] Following the procedure described in Example 17, Step 2,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-phenyl-ethyl ester,
[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester, and bis(triphenylphosphine)palladium(II)
dichloride were reacted to provide
{4'-[3-methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-3-yl}-acetic acid ethyl ester, which was hydrolyzed to the acid
as described in Example 17, Step 3.
Example 49
Synthesis of
4'-[3-Methyl-4-(R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl--
4-carboxylic acid (Compound 44)
[0314] Following the procedure described in Example 17, Step 2,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-phenyl-ethyl ester and 4-ethoxycarbonylphenylboronic acid
were reacted to provide
4'-[3-methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-
-4-carboxylic acid ethyl ester, which was hydrolyzed to the acid as
described in Example 17, Step 3.
Example 50
Synthesis of
(4'-{4-[1-(2,6-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 32)
[0315] Following the procedure described in Example 36, Step
5,5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxyl-
ic acid and 1-(2,6-dichlorophenyl)ethanol were reacted to provide
(4'-{4-[1-(2,6-dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 51
Synthesis of
2-(4'-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-2'-methyl-biphenyl-4-yl)-propionic acid (Compound 33)
[0316] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-3-methyl-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-fluoro-phenyl)-ethyl ester and
2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionic
acid ethyl ester were reacted to provide
2-(4'-{4-[(R)-1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-2'-methyl-biphenyl-4-yl)-propionic acid ethyl ester, which
was hydrolyzed to the acid as described in Example 36, Step 7.
Example 52
Synthesis of
(4'-{4-[(S)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 34)
[0317] (S)-1-(2-Fluoro-phenyl)-ethanol was prepared according to
the procedure described in Example 35, Step 1 using
2'-fluoroacetophenone and (R)-(+)-2-methyl-CBS--
oxazaborolidine.
[0318] Following the procedure described in Example 36, Step
5,5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxyl-
ic acid and (S)-1-(2-fluoro-phenyl)-ethanol were reacted to provide
(4'-{4-[(S)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 53
Synthesis of
(4'-{4-[(S)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 35)
[0319] (S)-1-(2-Chloro-phenyl)-ethanol was prepared as described in
Example 35, Step 1 using 2'-chloroacetophenone and
(R)-(+)-2-methyl-CBS-oxazaborolidine.
[0320] [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(S)-1-(2-chloro-phenyl)-ethyl ester was prepared as described in
Example 36, Step 5 using
5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylic
acid and (S)-1-(2-chloro-phenyl)-ethanol.
[0321] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(S)-1-(2-chloro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[(S)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 54
Synthesis of
{4'-[4-(2-Chloro-benzyloxycarbonylamino)-3-methyl-isoxazol-5-yl]-biphenyl-
-4-yl}-acetic acid (Compound 36)
[0322] [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
2-chloro-benzyl ester was prepared as described in Example 36, Step
5 using 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
chlorobenzyl alcohol.
[0323] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
2-chloro-benzyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceti- c
acid ethyl ester were reacted to provide
{4'-[4-(2-chloro-benzyloxycarbonylamino)-3-methyl-isoxazol-5-yl]-biphenyl-
-4-yl}-acetic acid ethyl ester, which was hydrolyzed to the acid as
described in Example 36, Step 7.
Example 55
Synthesis of
(4'-{4-[1-(2,3-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 38)
[0324] 2',3'-Difluoroacetophenone (0.337 g, 2.16 mmol) in methanol
(15 mL) was treated with sodium borohydride (0.090 g, 2.37 mmol),
and the reaction was stirred at room temperature for 30 minutes.
The mixture was partitioned between dichloromethane and water, and
the organic layer was separated, dried over MgSO.sub.4, filtered,
and concentrated to give 1-(2,3-difluoro-phenyl)-ethanol.
[0325] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-(2,3-difluoro-phenyl)-ethanol were reacted as described in
Example 36, Step 5 to provide
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,3-difluoro-phenyl)-ethyl ester.
[0326] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,3-difluoro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[1-(2,3-difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 56
Synthesis of
(4'-{4-[1-(2,4-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 39)
[0327] 2',4'-difluoroacetophenone was reduced to
1-(2,4-Difluorophenyl)ethanol as described in Example 55.
[0328] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-(2,4-difluorophenyl)ethanol were reacted as described in Example
36, Step 5 to provide
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,4-difluoro-phenyl)-ethyl ester.
[0329] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,4-difluoro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[1-(2,4-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 57
Synthesis of
(4'-{4-[1-(2-Fluoro-4-methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxa-
zol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 40)
[0330] 2'-Fluoro-4'-methoxyacetophenone was reduced to
1-(2-fluoro-4-methoxyphenyl)ethan-1-ol as described in Example
55.
[0331] 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-(2-fluoro-4-methoxyphenyl)ethan-1-ol were reacted as described in
Example 36, Step 5 to provide
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-fluoro-4-methoxy-phenyl)-ethyl ester.
[0332] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2-fluoro-4-methoxy-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[1-(2-fluoro-4-methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxa-
zol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 34, Step 2.
Example 58
Synthesis of
(4'-{4-[1-(2,5-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 41)
[0333] 2',5'-Difluoroacetophenone was reduced to
1-(2,5-Difluorophenyl)ethanol as described in Example 55.
[0334] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-(2,5-difluorophenyl)ethanol were reacted as described in Example
36, Step 5 to provide
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,5-difluoro-phenyl)-ethyl ester.
[0335] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,5-difluoro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[1-(2,5-difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 59
Synthesis of
(4'-{4-[1-(2,6-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid (Compound 42)
[0336] 2',6'-difluoroacetophenone was reduced to
1-(2,6-difluorophenyl)ethan-1-ol as described in Example 55.
[0337] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-(2,6-difluorophenyl)ethan-1-ol were reacted as described in
Example 36, Step 5 to provide
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,6-difluoro-phenyl)-ethyl ester.
[0338] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-(2,6-difluoro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[1-(2,6-difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 34, Step 2.
Example 60
Synthesis of
{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-2-yl}-acetic acid (Compound 45)
[0339] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-phenyl-ethyl ester and
[2-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-2-yl}-acetic acid ethyl ester, which was hydrolyzed to the acid
as described in Example 34, Step 2.
Example 61
Synthesis of
{4'-[3-Methyl-4-((R)-1-o-tolyl-ethoxycarbonylamino)-isoxazol-5-yl]hipheny-
l-4-yl}-acetic acid (Compound 46)
[0340] 2'-methylacetophenone and
(S)-(-)-2-methyl-CBS-oxazaborolidine were reacted as described in
Example 35, Step 1 to provide (R)-1-o-Tolyl-ethanol.
[0341] 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
(R)-1-o-tolyl-ethanol were reacted as described in Example 36, Step
5 to provide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-o-tolyl-ethyl ester.
[0342] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-o-tolyl-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
{4'-[3-methyl-4-((R)-1-o-tolyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphen-
yl-4-yl}-acetic acid ethyl ester, which was hydrolyzed to the acid
as described in Example 34, Step 2.
Example 62
Synthesis of
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-propionic acid (Diastereomers--Compound 47 and
Compound 48)
Step 1: 2-(4-Bromo-phenyl)-propionic Acid Ethyl Ester
[0343] 2-(4-Bromo-phenyl)-propionic acid ethyl ester (10.0 g, 41
mmol) was dissolved in tetrahydrofuran (400 mL) and cooled to
-78.degree. C. Sodium bis(trimethylsilyl)amide (2M in
tetrahydrofuran; 53.5 mL, 107 mmol) was added, and the mixture was
stirred for 1 hour at -78.degree. C. Iodomethane (2.814 mL, 45.1
mmol) was added, and the reaction was slowly warmed to room
temperature and stirred overnight. The mixture was concentrated,
and the residue was extracted with EtOAc. The combined organic
layers were washed with water and brine, and then dried and
concentrated. The crude material was purified by silica gel
chromatography (0-15% EtOAc in hexanes) to give the title
compound.
Step 2: 2-(4-Bromo-phenyl)-propionic Acid
[0344] Prepared according to the procedure described in Example 6,
Step 4 using 2-(4-bromo-phenyl)-propionic acid ethyl ester.
Step 3: 2-(4-Bromo-phenyl)-propionyl Chloride
[0345] 2-(4-Bromo-phenyl)-propionic acid (0.500 g, 2.18 mmol) in
dichloromethane (20 mL) was treated with dimethylformamide (0.03
mL, 0.44 mmol), followed by oxalyl chloride (0.248 mL, 2.8 mmol)
dropwise over 5 minutes, and the reaction was stirred at room
temperature for 1 hour. The mixture was then concentrated and dried
under vacuum to give the title compound.
Step 4:
(4R,5S)-3-[2-(4-Bromo-phenyl)-propionyl]-4-methyl-5-phenyl-oxazoli-
din-2-one
[0346] (4R,5S)-(+)-4-Methyl-5-phenyl-2-oxazolidinone (0.348 g, 1.96
mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to
-78.degree. C. n-Butyllithium (1.6M in hexanes; 1.64 mL, 2.62 mmol)
was added, and the reaction was stirred for 1 hour at -78.degree.
C. 2-(4-Bromo-phenyl)-propionyl chloride (0.540 g, 2.18 mmol) in
tetrahydrofuran (10 mL) was added, and the reaction was stirred for
30 minutes. The mixture was concentrated, and the residue was
partitioned between EtOAc and water. The organic layer was
separated, dried, and concentrated, and the crude material was
purified by silica gel chromatography (0-25% EtOAc in hexanes) to
give two diastereomers as separated products. Diastereomer A was
the first product to come off the column, while Diastereomer B was
the second product to come off the column.
Step 5:
(4R,5S)-4-Methyl-5-phenyl-3-{2-[4-(4,4,5,5-tetramethyl-[1,3,2]diox-
aborolan-2-yl)-phenyl]-propionyl}-oxazolidin-2-one
[0347] Diastereomer A--Prepared as described in Example 5, Step 2
using
(4R,5S)-3-[2-(4-bromo-phenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-o-
ne (Diastereomer A) and bis(pinacolato)diboron.
[0348] Diastereomer B--Prepared as described in Example 5, Step 2
using
(4R,5S)-3-[2-(4-bromo-phenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-o-
ne (Diastereomer B) and bis(pinacolato)diboron.
Step 6:
(3-Methyl-5-{4'-[1-methyl-2-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazo-
lidin-3-yl)-2-oxo-ethyl]-biphenyl-4-yl}-isoxazol-4-yl)-carbamic
acid (R)-1-(2-chloro-phenyl)-ethyl Ester
[0349] Diastereomer A--Prepared as described in Example 36, Step 6
using
(4R,5S)-4-methyl-5-phenyl-3-{2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborola-
n-2-yl)-phenyl]-propionyl}-oxazolidin-2-one (Diastereomer A) and
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester.
[0350] Diastereomer B--Prepared as described in Example 36, Step 6
using
(4R,5S)-4-methyl-5-phenyl-3-{2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborola-
n-2-yl)-phenyl]-propionyl}-oxazolidin-2-one (Diastereomer B) and
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester.
Step 7:
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-is-
oxazol-5-yl}-biphenyl-4-yl)-propionic Acid
[0351] Diastereomer A--Prepared as described in Example 34, Step 2
using
(3-methyl-5-{4'-[1-methyl-2-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-
-yl)-2-oxo-ethyl]-biphenyl-4-yl}-isoxazol-4-yl)-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester (Diastereomer A).
[0352] Diastereomer B--Prepared as described in Example 34, Step 2
using
(3-methyl-5-{4'-[1-methyl-2-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-
-yl)-2-oxo-ethyl]-biphenyl-4-yl}-isoxazol-4-yl)-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester (Diastereomer B)
Example 64
Synthesis of
(3'-Chloro-4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-i-
soxazol-5-yl}-biphenyl-4-yl)-acetic Acid (Compound 49)
[0353] Following the procedures described in Example 36:
4-bromo-2-chlorobenzoic acid and oxalyl chloride were reacted to
provide 4-bromo-2-chloro-benzoyl chloride, which was reacted with
3-methylamino-but-2-enoic acid methyl ester to provide
2-(4-bromo-2-chloro-benzoyl)-3-[(E)-methylimino]-butyric acid
methyl ester.
2-(4-Bromo-2-chloro-benzoyl)-3-[(E)-methylimino]-butyric acid
methyl ester and hydroxylamine hydrochloride were then reacted to
provide 5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic
acid methyl ester. Following Example 36, Step
4,5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid
methyl ester was hydrolyzed to
5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid.
5-(4-Bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid
and (R)-1-(2-chloro-phenyl)-ethanol were then reacted as described
in Example 36, Step 5 to provide
[5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester.
[0354] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(3'-chloro-4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-i-
soxazol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 36, Step 7.
Example 65
Synthesis of
2-(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol--
5-yl}-biphenyl-4-yl)-butyric Acid (Compound 50)
[0355] Prepared as described in Example 36, Step 6 using
{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-iso-
xazol-4-yl}-carbamic acid (R)-1-(2-chloro-phenyl)-ethyl ester and
2-(4-bromophenyl)butanoic acid.
Example 66
Synthesis of
(2'-Chloro-4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-i-
soxazol-5-yl}-biphenyl-4-yl)-acetic acid (Compound 51)
[0356] Following the procedures described in Example 36:
4-bromo-3-chlorobenzoic acid and oxalyl chloride were reacted to
provide 4-bromo-3-chloro-benzoyl chloride, which was then reacted
with 3-methylamino-but-2-enoic acid methyl ester to provide
2-(4-bromo-3-chloro-benzoyl)-3-[(E)-methylimino]-butyric acid
methyl ester.
2-(4-Bromo-3-chloro-benzoyl)-3-[(E)-methylimino]-butyric acid
methyl ester and hydroxylamine hydrochloride were then reacted as
described in Example 36, Step 3 to provide
5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid
methyl ester, which was then hydrolyzed to
5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid as
described in Example 36, Step 4.
5-(4-Bromo-3-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid
and (R)-1-(2-chloro-phenyl)-ethanol were reacted to provide
[5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester as described in Example 36,
Step 5.
[0357] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(2'-chloro-4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-i-
soxazol-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 36, Step 7.
Example 67
Synthesis of
(4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-2'-fluoro-biphenyl-4-yl)-acetic acid (Compound 52)
[0358] Following the procedures described in Example 36:
4-bromo-3-fluorobenzoyl chloride and oxalyl chloride were reacted
to provide 4-bromo-3-fluoro-benzoyl chloride, which was then
reacted with 3-methylamino-but-2-enoic acid methyl ester to provide
2-(4-bromo-3-fluoro-benzoyl)-3-[(E)-methylimino]-butyric acid
methyl ester.
2-(4-bromo-3-fluoro-benzoyl)-3-[(E)-methylimino]-butyric acid
methyl ester and hydroxylamine hydrochloride were then reacted as
described in Example 36, Step 3 to provide
5-(4-bromo-3-fluoro-phenyl)-3-methyl-isoxazole-4-carboxylic acid
methyl ester, which was then hydrolyzed to
5-(4-bromo-3-fluoro-phenyl)-3-methyl-isoxazole-4-carboxylic acid as
described in Example 36, Step 4.
5-(4-Bromo-3-fluoro-phenyl)-3-methyl-isoxazole-4-carboxylic acid
and (R)-1-(2-chloro-phenyl)-ethanol were reacted to provide
[5-(4-Bromo-3-fluoro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester as described in Example 36,
Step 5.
[0359] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-3-fluoro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5--
yl}-2'-fluoro-biphenyl-4-yl)-acetic acid ethyl ester, which was
hydrolyzed to the acid as described in Example 36, Step 7.
Example 68
Synthesis of
4'-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-y-
l}-biphenyl-4-carboxylic acid (Compound 53)
[0360] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(2-chloro-phenyl)-ethyl ester and
4-ethoxycarbonylphenylboronic acid were reacted to provide
4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-y-
l}-biphenyl-4-carboxylic acid ethyl ester, which was hydrolyzed to
the acid as described in Example 36, Step 7.
Example 71
Synthesis of
(4'-{4-[(R)-1-(3,5-Dibromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-
-5-yl}-biphenyl-4-yl)-acetic acid (Compound 56)
[0361] To a solution of 3,5-dibromobenzoic acid (2.5 g, 8.9 mmol)
in Et.sub.2O (30 mL) at 0.degree. C. was added methyl lithium (1.6M
in diethyl ether; 12.3 mL, 19.6 mmol) was added dropwise. The
reaction was warmed to room temperature and stirred for 2 hours.
Acidic work-up, followed by silica gel chromatography, gave
1-(3,5-dibromo-phenyl)-ethanone.
[0362] 1-(3,5-dibromo-phenyl)-ethanone and
(S)-(-)-2-methyl-CBS-oxazaborolidine were reacted as described in
Example 35, Step 1 to provide
(R)-1-(3,5-dibromo-phenyl)-ethanol.
[0363] Following the procedure described in Example 36, Step
5,5-(4'-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxyl-
ic acid and (R)-1-(3,5-dibromo-phenyl)-ethanol were reacted to
provide
(4'-{4-[(R)-1-(3,5-dibromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-
-5-yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed
to the acid as described in Example 34, Step 2.
Example 72
Synthesis of
{4'-[3-Methyl-4-((S)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bipheny-
l-4-yl}-acetic acid (Compound 57)
[0364] 2'-Chloroacetophenone and
(R)-(+)-2-methyl-CBS-oxazaborolidine were reacted as described in
Example 35, Step 1 to provide (S)-1-phenyl-ethanol.
[0365] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
(S)-1-phenyl-ethanol were reacted as described in Example 36, Step
5 to provide [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic
acid (S)-1-phenyl-ethyl ester.
[0366] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(S)-1-phenyl-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
{4'-[3-methyl-4-(S)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-
-4-yl}-acetic acid ethyl ester, which was hydrolyzed to the acid as
described in Example 34, Step 2.
Example 73
Synthesis of
(4'-{4-[(R)-1-(3-Hydroxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-
-yl}-biphenyl-4-yl)-acetic acid (Compound 58)
[0367] 3'-hydroxyacetophenone and
(S)-(-)-2-methyl-CBS-oxazaborolidine were reacted as described in
Example 35, Step 1 to provide 3-((R)-1-hydroxy-ethyl)-phenol.
[0368] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
3-((R)-1-hydroxy-ethyl)-phenol were reacted as described in Example
36, Step 5 to provide
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(3-hydroxy-phenyl)-ethyl ester.
[0369] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
(R)-1-(3-hydroxy-phenyl)-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl ester were reacted to provide
(4'-{4-[(R)-1-(3-hydroxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-
-yl}-biphenyl-4-yl)-acetic acid ethyl ester, which was hydrolyzed
to the acid as described in Example 34, Step 2.
Example 74
Synthesis of
{4'-[3-Methyl-4-(1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4--
yl}-acetic Acid (Compound 59)
[0370] 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-phenylethanol were reacted as described in Example 36, Step 5 to
provide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-phenyl-ethyl ester.
[0371] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-phenyl-ethyl ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceti- c
acid ethyl ester were reacted to provide
{4'-[3-methyl-4-(1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4--
yl}-acetic acid ethyl ester, which was hydrolyzed to the acid as
described in Example 34, Step 2.
Example 75
Synthesis of
{4'-[3-Methyl-4-(1-phenyl-ethoxy-d.sub.9-carbonylamino)-isoxazol-5-yl]-bi-
phenyl-4-yl}-acetic Acid (Compound 60)
[0372] 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and
1-phenylethanol-d.sub.9 (fully dueterated 1-phenylethanol obtained
from Carbocore) were reacted as described in Example 36, Step 5 to
provide [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-phenyl-ethyl-d9 ester.
[0373] Following the procedure described in Example 36, Step 6,
[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid
1-phenyl-ethyl-d9 ester and
[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic
acid ethyl were reacted to provide
{4'-[3-methyl-4-(1-phenyl-ethoxy-d9-carbonylamino)-isoxazol-5-yl]-bipheny-
l-4-yl}-acetic acid ethyl ester, which was hydrolyzed to the acid
as described in Example 34, Step 2.
Example 76
Synthesis of
[5-(4'-Carbamimidoylmethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbami-
c acid (R)-1-(2-fluoro-phenyl)-ethyl Ester (Compound 65)
[0374] Step 1: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-fluoro-phenyl)-ethyl ester and
4-cyanomethylphenylboronic acid were reacted as described in
Example 36, Step 6 to provide
[5-(4'-cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-fluoro-phenyl)-ethyl ester.
[0375] Step 2:
[5-(4'-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic
acid (R)-1-(2-fluoro-phenyl)-ethyl ester (0.400 g, 0.88 mmol) in
ethanol (5 mL) was treated with 4N HCl in 1,4-dioxane (5 mL), and
the reaction was stirred overnight at room temperature. The mixture
was concentrated to dryness, and then dissolved in 2M NH.sub.3 in
methanol. The reaction was stirred overnight at room temperature,
and then additional 2M NH.sub.3 in methanol was added and the
reaction was stirred at 40.degree. C. for 1.5 hours. The mixture
was concentrated, and the residue was purified by preparative HPLC
to give the title compound.
Example 77
Synthesis of
{5-[4'-(2-Acetylamino-2-imino-ethyl)-biphenyl-4-yl]-3-methyl-isoxazol-4-y-
l}-carbamic Acid (R)-1-(2-fluoro-phenyl)-ethyl Ester (Compound
66)
[0376] To
[5-(4'-carbamimidoylmethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl-
]-carbamic acid (R)-1-(2-fluoro-phenyl)-ethyl ester (0.055 g, 0.12
mmol) in dichloromethane (5 mL) was added diisopropylethylamine
(0.052 mL, 0.3 mmol), followed by acetyl chloride (0.009 mL, 0.126
mmol), and the reaction was stirred at room temperature. Additional
acetyl chloride (0.009 mL, 0.126 mmol) was added, and the reaction
was stirred at room temperature. The mixture was quenched with
water and extracted with dichloromethane. The combined organic
layers were dried, filterd, and concentrated, and the residue was
purified by preparative HPLC to give the title compound.
Example 78
Synthesis of
[4'-(4-Amino-3-methyl-isoxazol-5-yl)-biphenyl-4-yl]acetic acid
[0377]
[4'-(4-tert-Butoxycarbonylamino-3-methyl-isoxazol-5-yl)-biphenyl-4--
yl]-acetic acid (1.0 mmol) was treated with trifluoroacetic acid (5
mL) for 1 hour. Work-up provided the title compound. Mass spec.
data (M+H)=309.
Example 79
Synthesis of
2-(2-{4'-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-bi-
phenyl-4-yl}-acetylamino)-ethanesulfonic Acid (Compound 67)
[0378] Rats were dosed with
{4'-[3-ethyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-
-4-yl}-acetic acid (30 mg/kg) and the bile was collected over 6
hours. The title compound was purified from bile by reverse phase
HPLC. Mass spec. data (M+H)=564.
[0379] In some embodiments, Mass spectrometric data (mass spec.
data) is obtained on with a Shimadzu LCMS 2010A.
In Vitro Assay(s)
Example 80
Establishment of a CHO Cell Line Stably Expressing Human
LPA.sub.1
[0380] A 1.1 kb cDNA encoding the human LPA.sub.1 receptor was
cloned from human lung. Human lung RNA (Clontech Laboratories, Inc.
USA) was reverse transcribed using the RETROscript kit (Ambion,
Inc.) and the full-length cDNA for human LPA.sub.1 was obtained by
PCR of the reverse transcription reaction. The nucleotide sequence
of the cloned human LPA.sub.1 was determined by sequencing and
confirmed to be identical to the published human LPA.sub.1 sequence
(An et al. Biochem. Biophys. Res. Commun. 231:619 (1997). The cDNA
was cloned into the pcDNA5/FRT expression plasmid and transfected
in CHO cells using lipofectamine 2000 (Invitrogen Corp., USA).
Clones stably expressing human LPA.sub.1 were selected using
hygromycin and identified as cells that show Ca-influx in response
to LPA.
Example 81
Generation of Cells Transiently Expressing Human LPA.sub.2
[0381] A vector containing the human LPA.sub.2 receptor cDNA was
obtained from the Missouri S&T cDNA Resource Center
(www.cdna.org). The full-length cDNA fragment for human LPA.sub.2
was obtained by PCR from the vector. The nucleotide sequence of the
cloned human LPA.sub.2 was determined by sequencing and confirmed
to be identical to the published human LPA.sub.2 sequence (NCBI
accession number NM 004720). The cDNA was cloned into the pcDNA3.1
expression plasmid and transfected into B103 cells (Invitrogen
Corp., USA) by seeding cells in a 96-well poly-D-lysine coated
plate at 30,000-35,000 cells per well together with 0.2 .mu.l
lipofectamine 2000 and 0.2 .mu.g of the LPA.sub.2 expression
vector. Cells were cultured overnight in complete media before
being assayed for LPA-induced Ca-influx.
Example 82
Establishment of a CHO Cell Line Stably Expressing Human
LPA.sub.3
[0382] A vector containing the human LPA.sub.3 receptor cDNA was
obtained from the Missouri S&T cDNA Resource Center
(www.cdna.org). The full-length cDNA fragment for human LPA.sub.3
was obtained by PCR from the vector. The nucleotide sequence of the
cloned human LPA.sub.3 was determined by sequencing and confirmed
to be identical to the published human LPA.sub.3 sequence (NCBI
accession number NM 012152). The cDNA was cloned into the
pcDNA5/FRT expression plasmid and transfected in CHO cells using
lipofectamine 2000 (Invitrogen Corp., USA). Clones stably
expressing human LPA.sub.3 were selected using hygromycin and
identified as cells that show Ca-influx in response to LPA.
Example 83
LPA1 and LPA3 Calcium Flux Assays
[0383] Human LPA.sub.1 or LPA.sub.3 expressing CHO cells are seeded
at 20,000-45,000 cells per well in a 96-well poly-D-lysine coated
plate one or two days before the assay. Prior to the assay, the
cells are washed once with PBS and then cultured in serum-free
media overnight. On the day of the assay, a calcium indicator dye
(Calcium 4, Molecular Devices) in assay buffer (HBSS with Ca.sup.2+
and Mg.sup.2+ and containing 20 mM Hepes and 0.3% fatty-acid free
human serum albumin) is added to each well and incubation continued
for 1 hour at 37.degree. C. 10 .mu.l of test compounds in 2.5% DMSO
are added to the cells and incubation continued at room temperature
for 30 minutes. Cells are the stimulated by the addition of 10 nM
LPA and intracellular Ca.sup.2+ measured using the Flexstation 3
(Molecular Devices). IC.sub.50s are determined using Graphpad prism
analysis of drug titration curves.
Example 84
LPA2 Calcium Flux Assay
[0384] Following an overnight culture with lipofectamine 2000 and
the LPA.sub.2 expression vector, the B103 cells are washed once
with PBS then serum starved for 4 hours. A calcium indicator dye
(Calcium 4, Molecular Devices) in assay buffer (HBSS with Ca.sup.2+
and Mg.sup.2+ and containing 20 mM Hepes and 0.3% fatty-acid free
human serum albumin) is added to each well and incubation continued
for 1 hour at 37.degree. C. 10 .mu.l of test compounds in 2.5% DMSO
are added to the cells and incubation continued at room temperature
for 30 minutes. Cells are the stimulated by the addition of 10 nM
LPA and intracellular Ca.sup.2+ measured using the Flexstation 3
(Molecular Devices). IC.sub.50s are determined using Graphpad prism
analysis of drug titration curves.
Example 85
GTP.gamma.S Binding Assay
[0385] The ability of a compound to inhibit binding of GTP to
LPA.sub.1 is assessed via a membrane GTP.gamma.S assay. CHO cells
stably expressing the recombinant human LPA.sub.1 receptor are
resuspended in 10 mM Hepes, 7.4 containing 1 mM DTT, lysed and
centrifuged at 75,000.times.g to pellet the membranes. The
membranes are resuspended in 10 mM Hepes, 7.4 containing 1 mM DTT
and 10% glycerol. Membranes (.about.25 .mu.g per well) are
incubated in 96-well plates with 0.1 nM [.sup.35S]-GTP.gamma.S, 900
nM LPA, 5 .mu.M GDP, and test compound in Assay Buffer (50 mM
Hepes, pH 7.4, 100 mM NaCl, 10 mM MgCl.sub.2, 50 .mu.g/ml saponin
and 0.2% fatty-acid free human serum albumin) for 30 minutes at
30.degree. C. The reactions are terminated by rapid filtration
through Whatman GF/B glass fibre filter plates. The filter plates
are washed 3 times with 1 ml cold Wash Buffer (50 mM Hepes, 7.5,
100 mM NaCl and 10 mM MgCl.sub.2) and dried. Scintillant is then
added to the plates and the radioactivity retained on the filters
is determined on a Packard TopCount (Perkin Elmer). Specific
binding is determined as total radioactive binding minus
non-specific binding in the absence of the ligand (900 nM LPA).
IC.sub.50s were determined using Graphpad prism analysis of drug
titration curves.
[0386] Illustrative in vitro biological data for representative
compounds of Formula (I) is presented in the Table below. Unless
otherwise noted, compounds that were tested had an IC.sub.50 of
less than 50 .mu.M in the HLPA1 Ca Flux assay.
TABLE-US-00004 Compound HLPA1 Ca HLPA3 Ca Flux Number Flux IC50
(uM) IC50 (uM) 1 A C 2 B C 3 A C 4 A C 5 A C 6 A C 7 C C 8 A C 9 C
ND 10 A C 11 A C 12 A B 13 A C 14 A C 15 A C 16 A C 17 A ND 18 A C
19 A ND 20 A C 21 A ND 22 A C 23 A C 24 A C 25 A C 26 A 27 A C 28 A
B 29 A C 30 A C 31 A B 32 C ND 33 A ND 34 A C 35 B C 36 B ND 37 A C
38 A C 39 A C 40 C ND 41 A C 42 A D 43 B D 44 A C 45 C ND 46 A C 47
A B 48 A C 49 A C 50 A B 51 A C 52 A C 53 A B 56 C ND 57 A C 58 C D
61 A C 62 A D 63 A C 64 A C 65 C ND 66 A ND 67 A ND A = less than
0.3 .mu.M; B = greater than 0.3 .mu.M and less than 1 .mu.M; C =
greater than 1 .mu.M and less than 50 .mu.M; D = greater than 50
.mu.M; ND = not determined.
Example 86
LPA1 Chemotaxis Assay
[0387] Chemotaxis of the A2058 human melanoma cells was measured
using the Neuroprobe ChemoTx.RTM. System plates (8 .mu.m pore size,
5.7 mm diameter sites). The filter sites were coated with 0.001%
fibronectin (Sigma) in 20 mM Hepes, pH 7.4 and allowed to dry.
A2058 cells were serum-starved for 24 hours, then harvested with
Cell Stripper and resuspended in DMEM containing 0.1%
fatty-acid-free bovine serum albumin (BSA) to a concentration of
1.times.10.sup.6/ml. Cells were mixed with an equal volume of test
compound (2.times.) in DMEM containing 0.1% fatty-acid-free BSA and
incubated at 37.degree. C. for 15 minutes. LPA (100 nM in DMEM
containing 0.1% fatty-acid-free BSA) or vehicle was added to each
well of the lower chamber and 50 .mu.l of the cell suspension/test
compound mix was applied to the upper portion of the ChemoTx plate.
Plates were incubated at 37.degree. C. for three hours and then the
cells removed from the upper portion by rinsing with PBS and
scraping. The filter was dried then stained with HEMA 3 Staining
System (Fisher Scientific). The absorbance of the filter was read
at 590 nM and IC.sub.50s were determined using Symyx Assay
Explorer.
[0388] Compound 27 and Compound 37 inhibited LPA-driven chemotaxis
(IC.sub.50 less than 300 nM) of human A2058 melanoma cells.
In Vivo Assay(s)
Example 87
Bleomycin-Induced Lung Fibrosis Model in Mice
[0389] Female CD-1 mice (Harlan, 25-30 g) are housed 4 per cage,
given free access to food and water and allowed to acclimate for at
least 7 days prior to test initiation. After the habituation phase,
mice are lightly anesthetized with isoflurane (5% in 100% O.sub.2)
and administered with bleomycin sulfate (0.01-5 U/kg, Henry Schein)
via intratracheal instillation (Cuzzocrea S et al. Am J Physiol
Lung Cell Mol. Physiol. 2007 May; 292(5):L1095-104. Epub 2007 Jan.
12.). Mice are returned to their cages and monitored daily for the
duration of the experiment. Test compound or vehicle is delivered
po, ip or sc daily. The route and frequency of dosing is based on
previously determined pharmacokinetic properties. All animals are
sacrificed using inhaled isoflurane 3, 7, 14, 21 or 28 days after
bleomycin instillation. Following sacrifice, mice are intubated
with a 20 gauge angiocatheter attached to a 1 ml syringe. Lungs are
lavaged with saline to obtain bronchoalveolar lavage fluid (BALF)
and then removed and fixed in 10% neutral buffered formalin for
subsequent histopathological analysis. BALF is centrifuged for 10
min at 800.times.g to pellet the cells and the cell supernatant
removed and frozen at -80.degree. C. for subsequent protein
analysis using the DC protein assay kit (Biorad, Hercules, Calif.)
and soluble collagen analysis using Sircol (Biocolor Ltd, UK). BALF
is analyzed for concentrations of inflammatory, pro-fibrotic and
tissue injury biomarkers including transforming growth factor
.beta.1, hyaluronic acid, tissue inhibitor of metalloproteinase-1,
matrix matelloproteinase-7, connective tissue growth factor and
lactate dehydrogenase activity, using commercially available ELISA.
The cell pellet is re-suspended in PBS. Total cell counts are then
obtained using a Hemavet hematology system (Drew Scientific, Wayne,
Pa.) and differential cells counts are determined using Shandon
cytospin (Thermo Scientific, Waltham, Mass.). Lung tissue is
stained using hematoxylin and eosin (H&E) and trichrome and
lung fibrosisis determined by semiquantitative histopathological
scoring (Ashcroft T. et al. J. Clin. Path. 1988; 41; 4, 467-470.)
using light microscopy (10.times. magnification) and quantitative,
computer-assisted densitometry of collagen in lung tissue sections
using light microscopy. The data are plotted using Graphpad prism
and statistical differences between groups determined.
[0390] In the acute setting (3 day), Compound 27 significantly
reduced total protein, lactate dehydrogenase activity (LDH; tissue
injury marker) and tissue inhibitor of metalloproteinase-1 (TIMP-1;
pro-fibrotic marker) concentrations in broncheoalveolar lavage
fluid (BALF). In the chronic setting (14 and 28 day) model,
Compound 27 maintained mouse body weight and decreased inflammatory
cell influx and fibrosis after a single bleomycin (1.5 units/kg)
instillation and decreased pulmonary resistance and lung fibrosis
following repeated bleomycin (3.0-5.0 units/kg/week)
instillations.
[0391] Compound 37 reduced total protein, lactate and TIMP-1 in the
BALF in the acute setting (3-day). Compound 37 decreased
inflammatory cell influx and fibrosis after a single bleomycin
instillation (3.0 units) in the chronic setting (14-days only).
Example 88
Mouse Carbon Tetrachloride (CCl.sub.4)-Induced Liver Fibrosis
Model
[0392] Female C57BL/6 mice (Harlan, 20-25 g) housed 4/cage are
given free access to food and water and allowed to acclimate for at
least 7 days prior to test initiation. After the habituation phase,
mice receive CCl.sub.4 (0.5-1.0 ml/kg body weight) diluted in corn
oil vehicle (100 .mu.L volume) via i.p. injection twice a week for
4-6 weeks. (Higazi, A. A. et al., Clin Exp Immunol. 2008 April;
152(1):163-73. Epub 2008 Feb. 14.). Control mice receive an
equivalent volume of corn oil vehicle only. Test compound or
vehicle is delivered po, ip or sc daily. At the end of the study
(4-6 weeks after first i.p. injection of CCl.sub.4), mice are
sacrificed using inhaled isoflurane and blood is drawn via cardiac
puncture for subsequent analysis of ALT/AST levels. The liver is
harvested, and one half of the liver is frozen at -80.degree. C.
and the other half is fixed in 10% neutral buffered formalin for
histological assessment of liver fibrosis using light microscopy
(10.times. magnification). Liver tissue homogenates are analyzed
for collagen levels using Sircol (Biocolor Ltd, UK). Fixed Liver
tissue is stained using hematoxylin and eosin (H&E) and
trichrome and liver fibrosis is determined by quantitative,
computer-assisted densitometry of collagen in liver tissue sections
using light microscopy. Plasma and liver tissue lysates are also
analyzed for concentrations of inflammatory, pro-fibrotic and
tissue injury biomarkers including transforming growth factor
.beta.1, hyaluronic acid, tissue inhibitor of metalloproteinase-1,
matrix matelloproteinase-7, connective tissue growth factor and
lactate dehydrogenase activity, using commercially available ELISA.
The resulting data are plotted using Graphpad prism and statistical
differences between groups determined.
[0393] In this experiment, Compound 27 significantly reduced
collagen deposition in the liver as compared to the untreated
group. Compound 27 (30 mg/kg, po, qd) had the same effect on
collagen deposition as pirfenidone. Compound 37 significantly
reduced collagen deposition in the liver as compared to untreated
control group.
Example 89
Mouse Intravenous LPA-Induced Histamine Release
[0394] A mouse intravenous LPA-induced histamine release model is
utilized to determine the in vivo potency of LPA.sub.1 and
LPA.sub.3 receptor antagonists. Female CD-1 mice (weighing 25-35
grams) are administered compound (i.p., s.c. or p.o.) in a volume
of 10 ml/kg 30 minutes to 24 hours prior to intravenous LPA
challenge (300 .mu.g/mouse in 0.1% FAF BSA). Immediately following
LPA challenge mice are placed into an enclosed Plexiglas chamber
and exposed to an isoflurane for a period of 2 minutes. They are
removed, decapitated and trunk blood collected into tubes
containing EDTA. Blood is then centrifuged at 10,000.times.g for 10
minutes at 4.degree. C. Histamine concentrations in the plasma are
determined by EIA. Drug concentrations in plasma are determined by
mass spectrometry. The dose to achieve 50% inhibition of blood
histamine release is calculated by nonlinear regression (Graphpad
Prism) and plotted as the ED.sub.50. The plasma concentration
associated with this dose is plotted as the EC.sub.50.
Example 90
Mouse Unilateral Ureteral Obstruction Kidney Fibrosis Model
[0395] Female C57BL/6 mice (Harlan, 20-25 g) housed 4/cage will be
given free access to food and water and allowed to acclimate for at
least 7 days prior to test initiation. After the habituation phase,
mice undergo unilateral ureteral obstruction (UUO) surgery or sham
to left kidney. Briefly, a longitudinal, upper left incision is
performed to expose the left kidney. The renal artery is located
and 6/0 silk thread is passed between the artery and the ureter.
The thread is looped around the ureter and knotted 3 times insuring
full ligation of ureter. The kidney is returned to abdomen, the
abdominal muscle is sutured and the skin is stapled closed. Mice
are returned to their cages and monitored daily for the duration of
the experiment. Test compound or vehicle is delivered po, ip or sc
daily. The route and frequency of dosing is based on previously
determined pharmacokinetic properties. All animals are sacrificed
using inhaled isoflurane 4, 8 or 14 days after UUO surgery.
Following sacrifice blood is drawn via cardiac puncture, the
kidneys are harvested and one half of the kidney is frozen at
-80.degree. C. and the other half is fixed in 10% neutral buffered
formalin for histological assessment of kidney fibrosis using light
microscopy (10.times. magnification). Kidney tissue homogenates are
analyzed for collagen levels using Sircol (Biocolor Ltd, UK). Fixed
kidney tissue is also stained using hematoxylin and eosin (H&E)
and trichrome and kidney fibrosis is determined by quantitative,
computer-assisted densitometry of collagen in liver tissue sections
using light microscopy. Plasma and kidney tissue lysates are also
analyzed for concentrations of inflammatory, pro-fibrotic and
tissue injury biomarkers including transforming growth factor
.beta.1, hyaluronic acid, tissue inhibitor of metalloproteinase-1,
matrix matelloproteinase-7, connective tissue growth factor and
lactate dehydrogenase activity, using commercially available ELISA.
The resulting data are plotted using Graphpad prism and statistical
differences between groups determined.
[0396] In this experiment, Compound 27 reduced kidney fibrosis by
at least 20% as compared to the untreated group. Compound 37
reduced kidney fibrosis by 55% as compared to untreated group.
Example 91
Clinical Trial in Humans with Idiopathic Pulmonary Fibrosis (IPF)
Purpose
[0397] The purposes of this study is to assess the efficacy of
treatment with a compound of Formula (I) compared with placebo in
patients with idiopathic pulmonary fibrosis (IPF) and to assess the
safety of treatment with a compound of Formula (I) compared with
placebo in patients with IPF.
[0398] The primary outcome variable is the absolute change in
percent predicted forced vital capacity (FVC) from baseline to Week
72.
[0399] Secondary outcome measures include: composite outcomes of
important IPF-- related events; progression-free survival;
categorical assessment of absolute change in percent predicted FVC
from baseline to Week 72; change in Shortness-of-Breath from
baseline to Week 72; change in percent predicted hemoglobin
(Hb)-corrected carbon monoxide diffusing capacity (DLco) of the
lungs from baseline to Week 72; change in oxygen saturation during
the 6 minute walk test (6MWT) from baseline to Week 72; change in
high-resolution computed tomography (HRCT) assessment from baseline
to Week 72; change in distance walked in the 6MWT from baseline to
Week 72.
Criteria
[0400] Patients eligible for this study include those patients that
satisfy the following inclusion criteria: diagnosis of IPF; 40 to
80 years of age; FVC.gtoreq.50% predicted value; DLco.gtoreq.35%
predicted value; either FVC or DLco.ltoreq.90% predicted value; no
improvement in past year; able to walk 150 meters in 6 minutes and
maintain saturation .gtoreq.83% while on no more than 6 L/min
supplemental oxygen.
[0401] Patients are excluded from this study if they satisfy any of
the following criteria: unable to undergo pulmonary function
testing; evidence of significant obstructive lung disease or airway
hyper-responsiveness; in the clinical opinion of the investigator,
the patient is expected to need and be eligible for a lung
transplant within 72 weeks of randomization; active infection;
liver disease; cancer or other medical condition likely to result
in death within 2 years; diabetes; pregnancy or lactation;
substance abuse; personal or family history of long QT syndrome;
other IPF treatment; unable to take study medication; withdrawal
from other IPF trials.
[0402] Patients are orally dosed with either placebo or an amount
of compound of Formula (I) (1 mg/day-1000 mg/day). The primary
outcome variable will be the absolute change in percent predicted
FVC from Baseline to Week 72. Patients will receive blinded study
treatment from the time of randomization until the last patient
randomized has been treated for 72 weeks. A Data Monitoring
Committee (DMC) will periodically review safety and efficacy data
to ensure patient safety.
[0403] After week 72, patients who meet the Progression of Disease
(POD) definition, which is a .gtoreq.10% absolute decrease in
percent predicted FVC or a .gtoreq.15% absolute decrease in percent
predicted DLco, will be eligible to receive permitted IPF therapies
in addition to their blinded study drug. Permitted IPF therapies
include corticosteroids, azathioprine, cyclophosphamide and
N-acetyl-cysteine.
Example 92a
Parenteral Pharmaceutical Composition
[0404] To prepare a parenteral pharmaceutical composition suitable
for administration by injection (subcutaneous, intravenous, and the
like), 100 mg of a water-soluble salt of a compound of Formula (I)
is dissolved in sterile water and then mixed with 10 mL of 0.9%
sterile saline. The mixture is incorporated into a dosage unit form
suitable for administration by injection
[0405] In another embodiment, the following ingredients are mixed
to form an injectable formulation: 1.2 g of a compound of Formulas
(I), 2.0 mL of sodium acetate buffer solution (0.4 M), HCl (1 N) or
NaOH (1 M) (q.s. to suitable pH), water (distilled, sterile)
(q.s.to 20 mL). All of the above ingredients, except water, are
combined and stirred and if necessary, with slight heating if
necessary. A sufficient quantity of water is then added.
Example 92b
Oral Pharmaceutical Composition
[0406] To prepare a pharmaceutical composition for oral delivery,
100 mg of a compound of Formula (I) is mixed with 750 mg of starch.
The mixture is incorporated into an oral dosage unit for, such as a
hard gelatin capsule, which is suitable for oral
administration.
Example 92c
Sublingual (Hard Lozenge) Pharmaceutical Composition
[0407] To prepare a pharmaceutical composition for buccal delivery,
such as a hard lozenge, mix 100 mg of a compound of Formula (I)
with 420 mg of powdered sugar mixed, with 1.6 mL of light corn
syrup, 2.4 mL distilled water, and 0.42 mL mint extract. The
mixture is gently blended and poured into a mold to form a lozenge
suitable for buccal administration.
Example 92d
Fast-Disintegrating Sublingual Tablet
[0408] A fast-disintegrating sublingual tablet is prepared by
mixing 48.5% by weigh of a compound of Formula (I), 44.5% by weight
of microcrystalline cellulose (KG-802), 5% by weight of
low-substituted hydroxypropyl cellulose (50 .mu.m), and 2% by
weight of magnesium stearate. Tablets are prepared by direct
compression (AAPS PharmSciTech. 2006; 7(2):E41). The total weight
of the compressed tablets is maintained at 150 mg. The formulation
is prepared by mixing the amount of compound of Formula (I) with
the total quantity of microcrystalline cellulose (MCC) and
two-thirds of the quantity of low-substituted hydroxypropyl
cellulose (L-HPC) by using a three dimensional manual mixer
(lnversina.RTM., Bioengineering AG, Switzerland) for 4.5 minutes.
All of the magnesium stearate (MS) and the remaining one-third of
the quantity of L-HPC are added 30 seconds before the end of
mixing.
Example 92e
Inhalation Pharmaceutical Composition
[0409] To prepare a pharmaceutical composition for inhalation
delivery, 20 mg of a compound of Formula (I) is mixed with 50 mg of
anhydrous citric acid and 100 mL of 0.9% sodium chloride solution.
The mixture is incorporated into an inhalation delivery unit, such
as a nebulizer, which is suitable for inhalation
administration.
[0410] In another embodiment, compound of Formula (I) (500 mg) is
suspended in sterile water (100 mL), Span 85 (1 g) is added
followed by addition of dextrose (5.5 g) and ascorbic acid (10 mg).
Benzalkonium chloride (3 mL of a 1:750 aqueous solution) is added
and the pH is adjusted to 7 with phosphate buffer. The suspension
is packaged in sterile nebulizers.
Example 92f
Rectal Gel Pharmaceutical Composition
[0411] To prepare a pharmaceutical composition for rectal delivery,
100 mg of a compound of Formula (I) is mixed with 2.5 g of
methylcelluose (1500 mPa), 100 mg of methylparapen, 5 g of glycerin
and 100 mL of purified water. The resulting gel mixture is then
incorporated into rectal delivery units, such as syringes, which
are suitable for rectal administration.
Example 92g
Topical Gel Pharmaceutical Composition
[0412] To prepare a pharmaceutical topical gel composition, 100 mg
of a compound of Formula (I) is mixed with 1.75 g of hydroxypropyl
celluose, 10 mL of propylene glycol, 10 mL of isopropyl myristate
and 100 mL of purified alcohol USP. The resulting gel mixture is
then incorporated into containers, such as tubes, which are
suitable for topicl administration.
Example 92h
Ophthalmic Solution
[0413] To prepare a pharmaceutical opthalmic solution composition,
100 mg of a compound of Formula (I) is mixed with 0.9 g of NaCl in
100 mL of purified water and filterd using a 0.2 micron filter. The
resulting isotonic solution is then incorporated into ophthalmic
delivery units, such as eye drop containers, which are suitable for
ophthalmic administration.
Example 92i
Nasal Spray Solution
[0414] To prepare a pharmaceutical nasal spray solution, 10 g of a
compound of Formula (I) is mixed with 30 mL of a 0.05M phosphate
buffer solution (pH 4.4). The solution is placed in a nasal
administrator designed to deliver 100 .mu.l of spray for each
application.
[0415] The examples and embodiments described herein are for
illustrative purposes only and various modifications or changes
suggested to persons skilled in the art are to be included within
the spirit and purview of this application and scope of the
appended claims.
* * * * *