U.S. patent application number 14/272131 was filed with the patent office on 2014-08-21 for compounds and methods of treating ocular disorders.
The applicant listed for this patent is Case Western Reserve University. Invention is credited to Marcin Golczak, Akiko Maeda, Krzysztof Palczewski.
Application Number | 20140235604 14/272131 |
Document ID | / |
Family ID | 44146158 |
Filed Date | 2014-08-21 |
United States Patent
Application |
20140235604 |
Kind Code |
A1 |
Palczewski; Krzysztof ; et
al. |
August 21, 2014 |
COMPOUNDS AND METHODS OF TREATING OCULAR DISORDERS
Abstract
A method of treating an ocular disorder in a subject associated
with aberrant all-trans-retinal clearance in the retina, the method
comprising administering to the subject a therapeutically effective
amount of a primary amine compound of formula: ##STR00001## wherein
R.sub.1 is an aliphatic and/or aromatic compound.
Inventors: |
Palczewski; Krzysztof;
(Cleveland, OH) ; Maeda; Akiko; (Cleveland,
OH) ; Golczak; Marcin; (Cleveland, OH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Case Western Reserve University |
Cleveland |
OH |
US |
|
|
Family ID: |
44146158 |
Appl. No.: |
14/272131 |
Filed: |
May 7, 2014 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
13492193 |
Jun 8, 2012 |
8722669 |
|
|
14272131 |
|
|
|
|
PCT/US10/59426 |
Dec 8, 2010 |
|
|
|
13492193 |
|
|
|
|
61267645 |
Dec 8, 2009 |
|
|
|
Current U.S.
Class: |
514/210.1 ;
514/249; 514/380; 514/562; 514/563; 514/567; 514/640; 514/646;
514/659 |
Current CPC
Class: |
C07C 237/48 20130101;
A61K 31/428 20130101; A61K 31/4162 20130101; A61K 31/4184 20130101;
C07C 229/60 20130101; C07C 317/36 20130101; C07C 229/64 20130101;
A61P 27/02 20180101; A61K 9/0048 20130101; A61K 31/56 20130101;
C07C 211/01 20130101; C07D 261/04 20130101; A61K 31/197 20130101;
C07C 229/12 20130101; C07C 251/58 20130101; C07D 487/04
20130101 |
Class at
Publication: |
514/210.1 ;
514/249; 514/380; 514/562; 514/563; 514/567; 514/640; 514/646;
514/659 |
International
Class: |
C07D 487/04 20060101
C07D487/04; C07C 229/12 20060101 C07C229/12; C07C 237/48 20060101
C07C237/48; C07C 229/64 20060101 C07C229/64; C07C 251/58 20060101
C07C251/58; C07C 317/36 20060101 C07C317/36; C07C 211/01 20060101
C07C211/01; C07D 261/04 20060101 C07D261/04; C07C 229/60 20060101
C07C229/60 |
Goverment Interests
GOVERNMENT FUNDING
[0002] This invention was made with government support under Grant
No. EY09339 awarded by The National Institute of Health. The United
States Government has certain rights in the invention.
Claims
1-22. (canceled)
23. A method of treating a retinal pathology associated with
aberrant all-trans-retinal clearance in the retina outer segment of
a subject in need thereof, the method comprising: administering to
the subject a therapeutically effective amount of a primary amine
compound of formula: ##STR00045## wherein R.sub.1 is an aliphatic
and/or aromatic compound; the primary amine compound upon
administration to the subject forming a reversible Schiff-base with
the all-trans-retinal without adversely affecting normal retinoid
cycle performance; the primary amine compound when administered to
a Rdh8.sup.-/-Abca4.sup.-/- mouse increasing the optical coherence
tomography score of the mouse to at least about 2.5 and increasing
11-cis-retinal amount at least about 30% in comparison to untreated
control animal, wherein the primary amine compound is not a local
anesthetic, which includes an aromatic amine, that demonstrates
sodium channel blockade when administered to the subject.
24. The method of claim 23, wherein R.sub.1 is an aliphatic
compound.
25. The method of claim 23, wherein R.sub.1 is an aromatic
compound.
26. The method of claim 23, the primary amine compound not
inhibiting RPE65 enzymatic activity or any other proteins involved
in retinoid metabolism in the eye of the subject.
27. The method of claim 23, the primary amine compounds reducing
the formation of A2E and/or retinal dimer in the subject's
retina.
28. The method of claim 23, the primary amine compound being
delivered to the subject by at least one of topical administration,
systemic administration, intravitreal injection, and intraocular
delivery.
29. The method of claim 23, the retinal pathology comprising at
least one of macular degeneration, Stargardt's disease, geographic
atrophy, and retinitis pigmentosa.
30. The method of claim 23, the primary amine compound not causing
night blindness in the subject.
31. The method of claim 23, the primary compound comprising the
formula: ##STR00046## wherein R.sub.2 is hydrogen or
(C.sub.1-C.sub.6) straight chain or branched unsubstituted or
substituted alkyl; R.sub.3 is straight or branched unsubstituted or
substituted alkyl of from 1 to 8 carbon atoms, straight or branched
alkenyl of from 2 to 8 carbon atoms, cycloalkyl of from 3 to 7
carbon atoms, alkoxy of from 1 to 6 carbon atoms, -alkylcycloalkyl,
-alkylalkoxy, -alkyl, OH, -alkylphenyl, -alkylphenoxy, -phenyl or
substituted phenyl; R.sub.4 is hydrogen or (C.sub.1-C.sub.6)
straight chain or branched unsubstituted or substituted alkyl, or
carboxyl; Ar is phenyl which is unsubstituted or substituted with
1-5 of R.sub.7, wherein R.sub.7 is independently selected from the
group consisting of: (1) halogen, (2) C.sub.1-6 alkyl, which is
linear or branched and is unsubstituted or substituted with 1-5
halogens, (3) OC.sub.1-6 alkyl, which is linear or branched and is
unsubstituted or substituted with 1-5 halogens, and (4) CN; X.sub.1
is selected from the group consisting of: (1) N, and (2) CR.sub.6;
R.sub.5 and R.sub.6 are independently selected from the group
consisting of: (1) hydrogen, (2) CN, (3) C.sub.1-10 alkyl, which is
linear or branched and which is unsubstituted or substituted with
1-5 halogens or phenyl, which is unsubstituted or substituted with
1-5 substituents independently selected from halogen, CN, OH,
R.sub.8, OR.sub.8, NHSO.sub.2R.sub.8, SO.sub.2R.sub.8, CO.sub.2H,
and CO.sub.2C.sub.1-6 alkyl, wherein the CO.sub.2C.sub.1-6 alkyl is
linear or branched, (4) phenyl which is unsubstituted or
substituted with 1-5 substituents independently selected from
halogen, CN, OH, R.sub.8, OR.sub.8, NHSO.sub.2R.sub.8,
SO.sub.2R.sub.8, CO.sub.2H, and CO.sub.2C.sub.1-6 alkyl, wherein
the CO.sub.2C.sub.1-6 alkyl is linear or branched, and (5) a 5- or
6-membered heterocycle which may be saturated or unsaturated
comprising 1-4 heteroatoms independently selected from N, S and O,
the heterocycle being unsubstituted or substituted with 1-3
substituents independently selected from oxo, OH, halogen,
C.sub.1-6 alkyl, and OC.sub.1-6 alkyl, wherein the C.sub.1-6 alkyl
and OC.sub.1-6 alkyl are linear or branched and optionally
substituted with 1-5 halogens; R.sub.8 is C.sub.1-6 alkyl, which is
linear or branched and which is unsubstituted or substituted with
1-5 groups independently selected from halogen, CO.sub.2H, and
CO.sub.2C.sub.1-6 alkyl, wherein the CO.sub.2C.sub.1-6 alkyl is
linear or branched; R.sub.9 and R.sub.10 may be the same or
different and are hydrogen, straight or branched alkyl of from one
to six carbon atoms, lower alkylaryl, lower alkenyl, phenyl,
CF.sub.3, hydroxy, lower alkoxy, lower alkylthio, lower
alkylsulphonyl, CF.sub.3O, at the six position halogen, nitro,
carboxy, lower alkoxycarbonyl, NR.sub.11R.sub.12CO,
NR.sub.11R.sub.12, R.sub.11CONR.sub.12, CN,
NR.sub.11R.sub.12SO.sub.2, wherein R.sub.11 and R.sub.12 may be the
same or different and are hydrogen, lower alkyl, or aryl; R.sub.9
and R.sub.10 may together form a carbocyclic or methylenedioxy
ring; R.sub.14 is cyano, cyanomethyl, methoxymethyl, or
ethoxymethyl; X.sub.2 is O, N(H), or S, het is a 5 or 6-membered
heterocycle, n is 0, 1, 2, or 3, and each D is an unbranched lower
alkyl group; U is a substituent selected from halogen atom; cyano;
lower alkyl wherein one or more hydrogen atoms on the lower alkyl
group are optionally substituted by groups selected from a halogen
atom, hydroxyl, carbamoyl, amino, aryl, and a monocyclic or
bicyclic heterocyclic group containing one or more hetero-atoms
selected from nitrogen, oxygen, and sulfur atoms; lower alkylthio
wherein one or more hydrogen atoms on the alkyl group are
optionally substituted by groups selected from a halogen atom,
hydroxyl, carbamoyl, amino, and aryl; lower alkylsulfonyl wherein
one or more hydrogen atoms on the alkyl group are optionally
substituted by groups selected from a halogen atom, hydroxyl,
carbamoyl, amino, and aryl; hydroxyl; lower alkoxy; formyl; lower
alkylcarbonyl; arylcarbonyl; carboxyl; lower alkoxycarbonyl;
carbamoyl; N-lower alkylcarbamoyl; N,N-di-lower alkylaminocarbonyl;
amino; N-lower alkylamino; N,N-di-lower alkylamino; formylamino;
lower alkylcarbonylamino; aminosulfonylamino; (N-lower
alkylamino)sulfonylamino; (N,N-di-lower alkylamino)sulfonylamino;
aryl, optionally substituted by groups selected from a halogen
atom, hydroxyl, carbamoyl, aryl and amino; and a monocyclic or
bicyclic heterocyclic group containing one or more hetero-atoms
selected from nitrogen, oxygen, and sulfur atoms; W, X, Y, and Z
are each, independently, N, S, O CU or CH, such that at least one
of W, X, Y, and Z is N; A is ##STR00047## D is unbranched lower
alkyl; R.sub.15 and R.sub.16 are each independently substituted or
unsubstituted C.sub.1, C.sub.2, C.sub.3, C.sub.4, O.sub.5, O.sub.6,
O.sub.7, or C.sub.8, straight chain alkyl, or substituted or
unsubstituted C.sub.3, O.sub.4, O.sub.5, O.sub.6, C.sub.7, or
C.sub.8, branched chain alkyl; L is a single bond or CH.sub.2; m is
0, 1, or 2; n is 0, 1, 2, 3, or 4; Y.sub.1 is --(CH.sub.2).sub.2--,
--(CH.sub.2).sub.3--, --CH.sub.2CH(CH.sub.3)-- or
--CH.sub.2C(CH.sub.3).sub.2--; R.sub.17 is aryl or heteroaryl;
R.sub.18 and R.sub.19 are each independently C.sub.1-C.sub.4 alkyl
or 2-methoxyethyl; R.sub.20 is hydrogen, C.sub.1-C.sub.4 alkyl,
2-(C.sub.1-C.sub.4 alkoxy)ethyl, cyclopropylmethyl, benzyl, or
--(CH.sub.2).sub.m1COR.sub.21 where m1 is 1, 2 or 3 and R.sub.21 is
hydroxy, C.sub.1-C.sub.4 alkoxy or --NR.sub.22 where R.sub.22
hydrogen or C.sub.1-C.sub.4 alkyl; R.sub.23 and R.sub.24 can be the
same or different and are hydrogen, methyl, or ethyl as well as
pharmaceutically acceptable salts thereof.
32. The method of claim 23 the primary amine compound being
selected from the group consisting of: ##STR00048## ##STR00049##
and pharmaceutically acceptable salts thereof.
33. The method of claim 23, wherein the primary amine compound is
selected from the group consisting of: ##STR00050## and
pharmaceutically acceptable salts thereof.
34. The method of claim 23, the primary amine compound comprising
the formula: ##STR00051## wherein R.sub.2 is hydrogen, straight or
branched alkyl of from 1 to 6 carbon atoms or phenyl; R.sub.3 is
straight or branched alkyl of from 1 to 8 carbon atoms, straight or
branched alkenyl of from 2 to 8 carbon atoms, cycloalkyl of from 3
to 7 carbon atoms, alkoxy of from 1 to 6 carbon atoms,
-alkylcycloalkyl, -alkylalkoxy, -alkyl OH-alkylphenyl,
-alkylphenoxy, -phenyl or substituted phenyl; and R.sub.4 is
hydrogen, and R.sub.2 is straight or branched alkyl of from 1 to 6
carbon atoms or phenyl when R.sub.3 is methyl, or a
pharmaceutically acceptable salt thereof.
35. The method of claim 23, wherein the primary amine compound is
selected from the group consisting of:
3-Aminomethyl-5-methylhexanoic acid;
3-Aminomethyl-5-methylheptanoic acid;
3-Aminomethyl-5-methyl-octanoic acid;
3-Aminomethyl-5-methyl-nonanoic acid;
3-Aminomethyl-5-methyl-decanoic acid;
3-Aminomethyl-5-methyl-undecanoic acid;
3-Aminomethyl-5-methyl-dodecanoic acid;
3-Aminomethyl-5-methyl-tridecanoic acid;
3-Aminomethyl-5-cyclopropyl-hexanoic acid;
3-Aminomethyl-5-cyclobutyl-hexanoic acid;
3-Aminomethyl-5-cyclopentyl-hexanoic acid;
3-Aminomethyl-5-cyclohexyl-hexanoic acid;
3-Aminomethyl-5-trifluoromethyl-hexanoic acid;
3-Aminomethyl-5-phenyl-hexanoic acid;
3-Aminomethyl-5-(2-chlorophenyl)-hexanoic acid;
3-Aminomethyl-5-(3-chlorophenyl)-hexanoic acid;
3-Aminomethyl-5-(4-chlorophenyl)-hexanoic acid;
3-Aminomethyl-5-(2-methoxyphenyl)-hexanoic acid;
3-Aminomethyl-5-(3-methoxyphenyl)-hexanoic acid;
3-Aminomethyl-5-(4-methoxyphenyl)-hexanoic acid;
3-Aminomethyl-5-(phenylmethyl)-hexanoic acid;
(S)-3-(Aminomethyl)-5-methylhexanoic acid;
(R)-3-(Aminomethyl)-5-methylhexanoic acid;
(3R,4S)-3-Aminomethyl-4,5-dimethyl-hexanoic acid;
3-Aminomethyl-4,5-dimethyl-hexanoic acid;
(3R,4S)-3-Aminomethyl-4,5-dimethyl-hexanoic acid MP;
(3S,4S)-3-Aminomethyl-4,5-dimethyl-hexanoic acid;
(3R,4R)-3-Aminomethyl-4,5-dimethyl-hexanoic acid MP;
3-Aminomethyl-4-isopropyl-hexanoic acid;
3-Aminomethyl-4-isopropyl-heptanoic acid;
3-Aminomethyl-4-isopropyl-octanoic acid;
3-Aminomethyl-4-isopropyl-nonanoic acid;
3-Aminomethyl-4-isopropyl-decanoic acid;
3-Aminomethyl-4-phenyl-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-ethoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-propoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-isopropoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-tert-butoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-fluoromethoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-fluoro-ethoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3,3,3-trifluoro-propoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-phenoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-chloro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-chloro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-chloro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-fluoro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-fluoro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-fluoro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-methoxy-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-methoxy-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-methoxy-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-hydroxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-methoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-ethoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-propoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-isopropoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-tert-butoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-fluoromethoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-fluoro-ethoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-(3,3,3-trifluoro-propoxy)-hexanoic
acid; (3S,5S)-3-Aminomethyl-5-methyl-6-phenoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-chloro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-chloro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-chloro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-fluoro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-fluoro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-fluoro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-methoxy-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-methoxy-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-methoxy-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl
6-(4-trifluoromethyl-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl
6-(3-trifluoromethyl-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl
6-(2-trifluoromethyl-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl 6-(4-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl 6-(3-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl 6-(2-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-benzyloxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-7-hydroxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-methoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-ethoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-propoxy-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-isopropoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-tert-butoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-fluoromethoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(2-fluoro-ethoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(3,3,3-trifluoro-propoxy)-heptanoi-c
acid; (3S,5S)-3-Aminomethyl-7-benzyloxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-phenoxy-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(4-chloro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(3-chloro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(2-chloro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(4-fluoro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(3-fluoro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(2-fluoro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(4-methoxy-phenoxy)-5-methyl-heptanoic
acid;
(3S,5S)-3-Aminomethyl-7-(3-methoxy-phenoxy)-5-methyl-heptanoic
acid;
(3S,5S)-3-Aminomethyl-7-(2-methoxy-phenoxy)-5-methyl-heptanoic
acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(4-trifluoromethyl-phenoxy)-heptan-oic
acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(3-trifluoromethyl-phenoxy)-heptan-
-oic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(2-trifluoromethyl-phenoxy)-he-
ptan-oic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(4-nitro-phenoxy)-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(3-nitro-phenoxy)-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(2-nitro-phenoxy)-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-phenyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-chloro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-chloro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-chloro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-methoxy-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-methoxy-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-methoxy-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-fluoro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-fluoro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-fluoro-phenyl)-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-7-phenyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(4-chloro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(3-chloro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(2-chloro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(4-methoxy-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(3-methoxy-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(2-methoxy-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(4-fluoro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(3-fluoro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(2-fluoro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-oct-7-enoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-non-8-enoic acid;
(E)-(3S,5S)-3-Aminomethyl-5-methyl-oct-6-enoic acid;
(Z)-(3S,5S)-3-Aminomethyl-5-methyl-oct-6-enoic acid;
(Z)-(3S,5S)-3-Aminomethyl-5-methyl-non-6-enoic acid;
(E)-(3S,5S)-3-Aminomethyl-5-methyl-non-6-enoic acid;
(E)-(3S,5R)-3-Aminomethyl-5-methyl-non-7-enoic acid;
(Z)-(3S,5R)-3-Aminomethyl-5-methyl-non-7-enoic acid;
(Z)-(3S,5R)-3-Aminomethyl-5-methyl-dec-7-enoic acid;
(E)-(3S,5R)-3-Aminomethyl-5-methyl-undec-7-enoic acid;
(3S,5S)-3-Aminomethyl-5,6,6-trimethyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5,6-dimethyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclopropyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclobutyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclopentyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclohexyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-nonanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-decanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-undecanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-dodecanoic acid;
(3S,5R)-3-Aminomethyl-5,9-dimethyl-decanoic acid;
(3S,5R)-3-Aminomethyl-5,7-dimethyl-octanoic acid;
(3S,5R)-3-Aminomethyl-5,8-dimethyl-nonanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclopropyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclobutyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclopentyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclohexyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclopropyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclobutyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclopentyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclohexyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclopropyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclobutyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclopentyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclohexyl-5-methyl-octanoic acid;
(3S,5S)-3-Aminomethyl-6-fluoro-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-7-fluoro-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-8-fluoro-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-9-fluoro-5-methyl-nonanoic acid;
(3S,5S)-3-Aminomethyl-7,7,7-trifluoro-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-8,8,8-trifluoro-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-8-phenyl-octanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-phenyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-7-phenyl-heptanoic acid; and
pharmaceutically acceptable salts thereof.
36. The method of claim 23, wherein the primary amine compound is
selected from the group consisting of
(S)-3-(Aminomethyl)-5-methylhexanoic acid,
(R)-3-(Aminomethyl)-5-methylhexanoic acid, and racemic mixtures
thereof.
37. The method of claim 23, wherein the primary amine compound
administered to the subject includes less than about 1% by weight
(S)-3-(Aminomethyl)-5-methylhexanoic acid and greater than about
99% by weight (R)-3-(Aminomethyl)-5-methylhexanoic acid.
38. The method of claim 23, wherein the primary amine compound
administered to the subject includes less than about 1% by weight
(R)-3-(Aminomethyl)-5-methylhexanoic acid and greater than about
99% by weight (S)-3-(Aminomethyl)-5-methylhexanoic acid.
Description
RELATED APPLICATION
[0001] This application is a Continuation-in-Part of
PCT/US2010/059426, filed Dec. 8, 2010, which claims priority from
U.S. Provisional Application No. 61/267,645, filed Dec. 8, 2009,
the subject matter of which are incorporated herein by reference in
their entirety.
TECHNICAL FIELD
[0003] This application relates to compounds and methods of
treating ocular and/or retinal disorders that are associated with
aberrant all-trans-retinal clearance in the retina, and more
particularly to compounds and methods of treating retinal
degeneration and/or retinal disorders using primary amine
compounds.
BACKGROUND
[0004] The retinoid (visual) cycle is a complex enzymatic pathway
essential for regeneration of the visual chromophore,
11-cis-retinal, a component of rhodopsin and cone opsins that
undergoes activation by light in vertebrate eyes. Maintaining
continuous vision and preserving the health of photoreceptors
requires an adequate continuing supply of this aldehyde so
vertebrates evolved the retinoid cycle to achieve this objective.
The pathway operates in both photoreceptor cells and the retinal
pigmented epithelium (RPE), converting all-trans-retinal back to
11-cis-retinal by several chemical transformations. Whereas the
classical vertebrate retinoid cycle contributes primarily to
regeneration of rhodopsin in rod cells, RPE65-based chromophore
production may also be important for cone function.
[0005] Inadequate availability and/or processing of vitamin A to
the visual chromophore, 11-cis-retinal can adversely affect
vertebrate rhodopsin regeneration and visual congenital or
progressive blindness in humans. Inactivation of non-redundant
enzymes of the retinoid cycle, e.g., either LRAT that esterifies
all-trans-retinol or the retinoid isomerase called RPE65, produces
Leber congenital amaurosis (LCA), a leading cause of inherited
childhood blindness. LCA is an autosomal recessive, early onset
severe retinal dystrophy that accounts for 5% of all inherited
retinal dystrophies. Insufficient vitamin A in the diet also can
lead to progressive deterioration of vision and ultimately
blindness, a major problem in underdeveloped countries.
[0006] Whereas inadequate 11-cis-retinal production leads to
congenital blindness in humans, accumulation of the photoisomerized
chromophore all-trans-retinal also can be detrimental. Such is the
case when this reactive aldehyde is not efficiently cleared from
the internal membranes of retinal outer segment discs. Clearance of
all-trans-retinal involves two steps: 1). Translocation of
all-trans-retinal across the photoreceptor disc membranes by
ATP-binding cassette transporter 4 (ABCA4), and 2). Reduction of
all-trans-retinal to all-trans-retinol by retinol dehydrogenase 8
(RDH8), expressed in the outer segments of photoreceptors, and by
RDH12 located in photoreceptor inner segments.
[0007] ABCA4, also known as ABCR or the rim protein, localizes to
the rim of photoreceptor discs and transfers all-trans-retinal from
the inside to the outside of disc membranes after it is released
from visual pigments. Mutations in ABCA4 can cause Stargardt
macular degeneration, cone-rod dystrophy, or recessive RP. Also,
heterozygous mutations in ABCA4 increase the risk of developing
age-related macular degeneration.
Di-retinoid-pyridinium-retinylethanolamine (A2E) and retinal dimer
(RALdi) conjugates are the major fluorophores of lipofuscins
produced from all-trans-retinal. Even in the presence of a
functional transporter, both A2E and RALdi can accumulate as a
consequence of aging together with light exposure and produce toxic
effects on RPE cells. Patients affected by age-related macular
degeneration, Stargardt disease with a disabled ABCA4 gene or other
retinal diseases associated with lipofuscin accumulation develop
retinal degeneration. ABCA4 mutations also are linked to a high
risk of AMD.
SUMMARY
[0008] This application relates to compounds and methods of
treating an ocular disorder in a subject associated with aberrant
all-trans-retinal clearance in the retina. The ocular disorder can
include, for example, retinal disorders, such as retinal
degeneration, macular degeneration, including age-related macular
degeneration, Stargardt disease, geographic atrophy, and retinitis
pigmentosa. The method of treating the ocular disorder in a subject
can include administering to the subject a therapeutically
effective amount of a primary amine compound of formula:
##STR00002##
[0009] wherein R.sub.1 is an aliphatic and/or aromatic compound.
The primary amine compound upon administration to the subject forms
a reversible Schiff-base with the all-trans-retinal without
adversely affecting normal retinoid cycle performance. The primary
amine compound when administered to a Rdh8.sup.-/-Abca4.sup.-/-
mouse increases the optical coherence tomography score of the
mouse, which reflects severity in retinal morphology, to at least
about 2.5 and increases 11-cis-retinal amount at least about 30% in
comparison to untreated control animal. The primary amine compound
is not a local anesthetic, which includes an aromatic amine that
demonstrates sodium channel blockade when administered to the
subject.
[0010] In an aspect of the application, the primary amine compound
does not inhibit RPE65 enzymatic activity or any other proteins
involved in retinoid metabolism in the eye of the subject. The
primary amine compounds can reduce the formation of A2E and/or
retinal dimer in the subject's retina and promote 11-cis-retinal
production in the subject. The primary amine compound does not
induce night blindness.
[0011] In another aspect of the application, the primary amine
compound can have a molecular weight less than about 500 and be
delivered to the subject by at least one of topical administration,
systemic administration, intravitreal injection, and/or intraocular
delivery. In one example, the primary amine can be provided in an
ocular preparation for sustained delivery.
BRIEF DESCRIPTION OF THE DRAWINGS
[0012] FIG. 1 is a schematic illustration of the visual cycle.
[0013] FIG. 2 is a schematic illustration of retinoid flow and
all-trans-retinal clearance in the visual cycle.
[0014] FIGS. 3A-B illustrate UV/Vis spectra for active and inactive
primary amine compounds in accordance with an aspect of the
application.
[0015] FIGS. 4A-B illustrate UV/Vis spectra for active and inactive
primary amine compounds in accordance with an aspect of the
application.
[0016] FIG. 5 illustrates UV/Vis spectra for an active primary
amine compound in accordance with an aspect of the invention.
[0017] FIG. 6 illustrates a chromatogram of HPLC separation of
retinoids.
[0018] FIG. 7 illustrates a MS/MS spectrum of ritinyl imine
fragments in eyes of mice.
[0019] FIG. 8 illustrates SD-OCT images of retinas from WT and
Rdh8.sup.-/-Abca4.sup.-/- mice.
[0020] FIG. 9 illustrates OCT images showing grading of
retinas.
DETAILED DESCRIPTION
[0021] For convenience, certain terms employed in the
specification, examples, and appended claims are collected here.
Unless defined otherwise, all technical and scientific terms used
herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this application belongs.
[0022] The articles "a" and "an" are used herein to refer to one or
to more than one (i.e., to at least one) of the grammatical object
of the article. By way of example, "an element" means one element
or more than one element.
[0023] The terms "comprise," "comprising," "include," "including,"
"have," and "having" are used in the inclusive, open sense, meaning
that additional elements may be included. The terms "such as",
"e.g.", as used herein are non-limiting and are for illustrative
purposes only. "Including" and "including but not limited to" are
used interchangeably.
[0024] The term "or" as used herein should be understood to mean
"and/or", unless the context clearly indicates otherwise.
[0025] It will be noted that the structure of some of the compounds
of the application include asymmetric (chiral) carbon atoms. It is
to be understood accordingly that the isomers arising from such
asymmetry are included within the scope of the invention, unless
indicated otherwise. Such isomers can be obtained in substantially
pure form by classical separation techniques and by
stereochemically controlled synthesis. The compounds of this
application may exist in stereoisomeric form, therefore can be
produced as individual stereoisomers or as mixtures.
[0026] The term "isomerism" refers to compounds that have identical
molecular formulae but that differ in the nature or the sequence of
bonding of their atoms or in the arrangement of their atoms in
space. Isomers that differ in the arrangement of their atoms in
space are termed "stereoisomers". Stereoisomers that are not mirror
images of one another are termed "diastereoisomers", and
stereoisomers that are non-superimposable mirror images are termed
"enantiomers", or sometimes optical isomers. A carbon atom bonded
to four nonidentical substituents is termed a "chiral center".
[0027] The term "chiral isomer" refers to a compound with at least
one chiral center. It has two enantiomeric forms of opposite
chirality and may exist either as an individual enantiomer or as a
mixture of enantiomers. A mixture containing equal amounts of
individual enantiomeric forms of opposite chirality is termed a
"racemic mixture". A compound that has more than one chiral center
has 2n-1 enantiomeric pairs, where n is the number of chiral
centers. Compounds with more than one chiral center may exist as
either an individual diastereomer or as a mixture of diastereomers,
termed a "diastereomeric mixture". When one chiral center is
present, a stereoisomer may be characterized by the absolute
configuration (R or S) of that chiral center. Absolute
configuration refers to the arrangement in space of the
substituents attached to the chiral center. The substituents
attached to the chiral center under consideration are ranked in
accordance with the Sequence Rule of Cahn, Ingold and Prelog. (Cahn
et al, Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahn et
al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc.
1951 (London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn,
J., Chem. Educ. 1964, 41, 116).
[0028] The term "geometric isomers" refer to the diastereomers that
owe their existence to hindered rotation about double bonds. These
configurations are differentiated in their names by the prefixes
cis and trans, or Z and E, which indicate that the groups are on
the same or opposite side of the double bond in the molecule
according to the Cahn-Ingold-Prelog rules.
[0029] Further, the structures and other compounds discussed in
this application include all atropic isomers thereof. "Atropic
isomers" are a type of stereoisomer in which the atoms of two
isomers are arranged differently in space. Atropic isomers owe
their existence to a restricted rotation caused by hindrance of
rotation of large groups about a central bond. Such atropic isomers
typically exist as a mixture, however as a result of recent
advances in chromatography techniques, it has been possible to
separate mixtures of two atropic isomers in select cases.
[0030] The terms "crystal polymorphs" or "polymorphs" or "crystal
forms" means crystal structures in which a compound (or salt or
solvate thereof) can crystallize in different crystal packing
arrangements, all of which have the same elemental composition.
Different crystal forms usually have different X-ray diffraction
patterns, infrared spectral, melting points, density hardness,
crystal shape, optical and electrical properties, stability and
solubility. Recrystallization solvent, rate of crystallization,
storage temperature, and other factors may cause one crystal form
to dominate. Crystal polymorphs of the compounds can be prepared by
crystallization under different conditions.
[0031] The term "derivative", refers to compounds that have a
common core structure, and are substituted with various groups as
described herein. For example, all of the compounds represented by
formula I are primary amines and have formula I as a common
core.
[0032] The term "bioisostere" refers to a compound resulting from
the exchange of an atom or of a group of atoms with another,
broadly similar, atom or group of atoms. The objective of a
bioisosteric replacement is to create a new compound with similar
biological properties to the parent compound. The bioisosteric
replacement may be physicochemically or topologically based.
Examples of carboxylic acid bioisosteres include acyl sulfonimides,
tetrazoles, sulfonates, and phosphonates. See, e.g., Patani and
LaVoie, Chem. Rev. 96, 3147-3176 (1996).
[0033] The phrases "parenteral administration" and "administered
parenterally" refer to modes of administration other than enteral
and topical administration, such as injections, and include,
without limitation, intravenous, intramuscular, intrapleural,
intravascular, intrapericardial, intraarterial, intrathecal,
intracapsular, intraorbital, intracardiac, intradermal,
intraperitoneal, transtracheal, subcutaneous, subcuticular,
intra-articular, subcapsular, subarachnoid, intraspinal and
intrastemal injection and infusion.
[0034] The term "treating" refers to inhibiting a disease, disorder
or condition in a subject, e.g., impeding its progress; and
relieving the disease, disorder or condition, e.g., causing
regression of the disease, disorder and/or condition. Treating the
disease or condition includes ameliorating at least one symptom of
the particular disease or condition, even if the underlying
pathophysiology is not affected.
[0035] The term "preventing" refers to stopping a disease, disorder
or condition from occurring in a subject, which may be predisposed
to the disease, disorder and/or condition but has not yet been
diagnosed as having it. Preventing a condition related to a disease
includes stopping the condition from occurring after the disease
has been diagnosed but before the condition has been diagnosed.
[0036] The term a "pharmaceutical composition" refers to a
formulation containing the disclosed compounds in a form suitable
for administration to a subject. The pharmaceutical composition can
be in bulk or in unit dosage form. The unit dosage form is any of a
variety of forms, including, for example, a capsule, an IV bag, a
tablet, a single pump on an aerosol inhaler, or a vial. The
quantity of active ingredient (e.g., a formulation of the disclosed
compound or salts thereof) in a unit dose of composition is an
effective amount and is varied according to the particular
treatment involved. One skilled in the art will appreciate that it
is sometimes necessary to make routine variations to the dosage
depending on the age and condition of the patient. The dosage will
also depend on the route of administration. A variety of routes are
contemplated, including oral, pulmonary, rectal, parenteral,
transdermal, subcutaneous, intravenous, intramuscular,
intraperitoneal, intranasal, and the like. Dosage forms for the
topical or transdermal administration of a compound of this
invention include powders, sprays, ointments, pastes, creams,
lotions, gels, solutions, patches and inhalants. In a preferred
embodiment, the active compound is mixed under sterile conditions
with a pharmaceutically acceptable carrier, and with any
preservatives, buffers, or propellants that are required.
[0037] The term "flash dose" refers to compound formulations that
are rapidly dispersing dosage forms.
[0038] The term "immediate release" refers to a release of compound
from a dosage form in a relatively brief period of time, generally
up to about 60 minutes. The term "modified release" is defined to
include delayed release, extended release, and pulsed release. The
term "pulsed release" is defined as a series of releases of drug
from a dosage form. The term "sustained release" or "extended
release" is defined as continuous release of a compound from a
dosage form over a prolonged period.
[0039] The phrase "pharmaceutically acceptable" refers to
compositions, polymers and other materials and/or dosage forms
which are, within the scope of sound medical judgment, suitable for
use in contact with the tissues of human beings and animals without
excessive toxicity, irritation, allergic response, or other problem
or complication, commensurate with a reasonable benefit/risk
ratio.
[0040] The phrase "pharmaceutically acceptable carrier" refers to
pharmaceutically acceptable materials, compositions or vehicles,
such as a liquid or solid filler, diluent, excipient, solvent or
encapsulating material, involved in carrying or transporting any
subject composition from one organ, or portion of the body, to
another organ, or portion of the body. Each carrier must be
"acceptable" in the sense of being compatible with the other
ingredients of a subject composition and not injurious to the
patient. In certain embodiments, a pharmaceutically acceptable
carrier is non-pyrogenic. Some examples of materials which may
serve as pharmaceutically acceptable carriers include: (1) sugars,
such as lactose, glucose and sucrose; (2) starches, such as corn
starch and potato starch; (3) cellulose, and its derivatives, such
as sodium carboxymethyl cellulose, ethyl cellulose and cellulose
acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc;
(8) excipients, such as cocoa butter and suppository waxes; (9)
oils, such as peanut oil, cottonseed oil, sunflower oil, sesame
oil, olive oil, corn oil and soybean oil; (10) glycols, such as
propylene glycol; (11) polyols, such as glycerin, sorbitol,
mannitol and polyethylene glycol; (12) esters, such as ethyl oleate
and ethyl laurate; (13) agar; (14) buffering agents, such as
magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16)
pyrogen-free water; (17) isotonic saline; (18) Ringer's solution;
(19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other
non-toxic compatible substances employed in pharmaceutical
formulations.
[0041] The compounds of the application are capable of further
forming salts. All of these forms are also contemplated within the
scope of the claims.
[0042] The phrase "pharmaceutically acceptable salt" of a compound
means a salt that is pharmaceutically acceptable and that possesses
the desired pharmacological activity of the parent compound. For
example, the salt can be an acid addition salt. One embodiment of
an acid addition salt is a hydrochloride salt
[0043] The pharmaceutically acceptable salts can be synthesized
from a parent compound that contains a basic or acidic moiety by
conventional chemical methods. Generally, such salts can be
prepared by reacting the free acid or base forms of these compounds
with a stoichiometric amount of the appropriate base or acid in
water or in an organic solvent, or in a mixture of the two;
generally, non-aqueous media like ether, ethyl acetate, ethanol,
isopropanol, or acetonitrile are preferred. Lists of salts are
found in Remington's Pharmaceutical Sciences, 18th ed. (Mack
Publishing Company, 1990). For example, salts can include, but are
not limited to, the hydrochloride and acetate salts of the
aliphatic amine-containing, hydroxylamine-containing, and
imine-containing compounds of the present invention.
[0044] It should be understood that all references to
pharmaceutically acceptable salts include solvent addition forms
(solvates) or crystal forms (polymorphs) as defined herein, of the
same salt.
[0045] The compounds described herein can also be prepared as
esters, for example pharmaceutically acceptable esters. For
example, a carboxylic acid function group in a compound can be
converted to its corresponding ester, e.g., a methyl, ethyl, or
other ester. Also, an alcohol group in a compound can be converted
to its corresponding ester, e.g., an acetate, propionate, or other
ester.
[0046] The compounds described herein can also be prepared as
prodrugs, for example pharmaceutically acceptable prodrugs. The
terms "pro-drug" and "prodrug" are used interchangeably herein and
refer to any compound, which releases an active parent drug in
vivo. Since prodrugs are known to enhance numerous desirable
qualities of pharmaceuticals (e.g., solubility, bioavailability,
manufacturing, etc.) the compounds of the present invention can be
delivered in prodrug form. Thus, the present application is
intended to cover prodrugs of the presently claimed compounds,
methods of delivering the same and compositions containing the
same. "Prodrugs" are intended to include any covalently bonded
carriers that release an active parent drug in vivo when such
prodrug is administered to a subject. Prodrugs the present
invention are prepared by modifying functional groups present in
the compound in such a way that the modifications are cleaved,
either in routine manipulation or in vivo, to the parent compound.
Prodrugs include compounds described herein wherein a hydroxy,
amino, sulfhydryl, carboxy, or carbonyl group is bonded to any
group that may be cleaved in vivo to form a free hydroxyl, free
amino, free sulftydryl, free carboxy or free carbonyl group,
respectively.
[0047] Examples of prodrugs include, but are not limited to, esters
(e.g., acetate, dialkylaminoacetates, formates, phosphates,
sulfates, and benzoate derivatives) and carbamates (e.g.,
N,N-dimethylaminocarbonyl) of hydroxy functional groups, ester
groups (e.g., ethyl esters, morpholinoethanol esters) of carboxyl
functional groups, N-acyl derivatives (e.g., N-acetyl) N-Mannich
bases, Schiff bases and enaminones of amino functional groups,
oximes, acetals, ketals and enol esters of ketone and aldehyde
functional groups in compounds of Formula I, and the like (e.g.,
Bundegaard, H. "Design of Prodrugs" p 1-92, Elesevier, New
York-Oxford (1985)).
[0048] The term "protecting group" refers to a grouping of atoms
that when attached to a reactive group in a molecule masks, reduces
or prevents that reactivity. Examples of protecting groups can be
found in Green and Wuts, Protective Groups in Organic Chemistry,
(Wiley, 2.sup.nd ed. 1991); Harrison and Harrison et al.,
Compendium of Synthetic Organic Methods, Vols. 1-8 (John Wiley and
Sons, 1971-1996); and Kocienski, Protecting Groups, (Verlag,
3.sup.rd ed. 2003).
[0049] The term "amine protecting group" refers to a functional
group that converts an amine, amide, or other nitrogen-containing
moiety into a different chemical group that is substantially inert
to the conditions of a particular chemical reaction. Amine
protecting groups can be removed easily and selectively in good
yield under conditions that do not affect other functional groups
of the molecule. Examples of amine protecting groups include, but
are not limited to, formyl, acetyl, benzyl, t-butyldimethylsilyl,
t-butdyldiphenylsilyl, t-butyloxycarbonyl (Boc), p-methoxybenzyl,
methoxymethyl, tosyl, trifluoroacetyl, trimethylsilyl (TMS),
fluorenyl-methyloxycarbonyl, 2-trimethylsilyl-ethyoxycarbonyl,
1-methyl-1-(4-biphenylyl)ethoxycarbonyl, allyloxycarbonyl,
benzyloxycarbonyl (CBZ), 2-trimethylsilyl-ethanesulfonyl (SES),
trityl and substituted trityl groups, 9-fluorenylmethyloxycarbonyl
(FMOC), nitro-veratryloxycarbonyl (NVOC), and the like. Other amine
protecting groups can be identified by those of skill in the
art.
[0050] Representative hydroxy protecting groups include those where
the hydroxy group is either acylated or alkylated such as benzyl,
and trityl ethers as well as alkyl ethers, tetrahydropyranyl
ethers, trialkylsilyl ethers and allyl ethers.
[0051] Additionally, the salts of the compounds described herein,
can exist in either hydrated or unhydrated (the anhydrous) form or
as solvates with other solvent molecules. Nonlimiting examples of
hydrates include monohydrates, dihydrates, etc. Nonlimiting
examples of solvates include ethanol solvates, acetone solvates,
etc.
[0052] The term "solvates" refers to solvent addition forms that
contain either stoichiometric or non stoichiometric amounts of
solvent. Some compounds have a tendency to trap a fixed molar ratio
of solvent molecules in the crystalline solid state, thus forming a
solvate. If the solvent is water, the solvate formed is a hydrate;
when the solvent is alcohol, the solvate formed is an alcoholate.
Hydrates are formed by the combination of one or more molecules of
water with one of the substances in which the water retains its
molecular state as H.sub.2O, such combination being able to form
one or more hydrate.
[0053] The compounds, salts and prodrugs described herein can exist
in several tautomeric forms, including the enol and imine form, and
the keto and enamine form and geometric isomers and mixtures
thereof. All such tautomeric forms are included within the scope of
the present invention. Tautomers exist as mixtures of a tautomeric
set in solution. In solid form, usually one tautomer predominates.
Even though one tautomer may be described, the present application
includes all tautomers of the present compounds. A tautomer is one
of two or more structural isomers that exist in equilibrium and are
readily converted from one isomeric form to another. This reaction
results in the formal migration of a hydrogen atom accompanied by a
switch of adjacent conjugated double bonds. In solutions where
tautomerization is possible, a chemical equilibrium of the
tautomers will be reached. The exact ratio of the tautomers depends
on several factors, including temperature, solvent, and pH. The
concept of tautomers that are interconvertable by tautomerizations
is called tautomerism.
[0054] Of the various types of tautomerism that are possible, two
are commonly observed. In keto-enol tautomerism a simultaneous
shift of electrons and a hydrogen atom occurs.
[0055] Tautomerizations can be catalyzed by: Base: 1.
deprotonation; 2. formation of a delocalized anion (e.g. an
enolate); 3. protonation at a different position of the anion;
Acid: 1. protonation; 2. formation of a delocalized cation; 3.
deprotonation at a different position adjacent to the cation.
[0056] The term "analog" refers to a chemical compound that is
structurally similar to another but differs slightly in composition
(as in the replacement of one atom by an atom of a different
element or in the presence of a particular functional group, or the
replacement of one functional group by another functional group).
Thus, an analog is a compound that is similar or comparable in
function and appearance, but not in structure or origin to the
reference compound.
[0057] A "patient," "subject," or "host" to be treated by the
subject method may mean either a human or non-human animal, such as
primates, mammals, and vertebrates.
[0058] The term "prophylactic or therapeutic" treatment refers to
administration to the host of one or more of the subject
compositions. If it is administered prior to clinical manifestation
of the unwanted condition (e.g., disease or other unwanted state of
the host animal) then the treatment is prophylactic, i.e., it
protects the host against developing the unwanted condition,
whereas if it is administered after manifestation of the unwanted
condition, the treatment is therapeutic (i.e., it is intended to
diminish, ameliorate, or stabilize the existing unwanted condition
or side effects thereof).
[0059] The terms "therapeutic agent", "drug", "medicament" and
"bioactive substance" refer to molecules and other agents that are
biologically, physiologically, or pharmacologically active
substances that act locally or systemically in a patient or subject
to treat a disease or condition, such as retinal degeneration or
other forms of retinal disease whose etiology involves aberrant
clearance of all trans-retinal. The terms include without
limitation pharmaceutically acceptable salts thereof and prodrugs.
Such agents may be acidic, basic, or salts; they may be neutral
molecules, polar molecules, or molecular complexes capable of
hydrogen bonding; they may be prodrugs in the form of ethers,
esters, amides and the like that are biologically activated when
administered into a patient or subject.
[0060] The phrase "therapeutically effective amount" is an
art-recognized term. In certain embodiments, the term refers to an
amount of a therapeutic agent that, when incorporated into a
polymer, produces some desired effect at a reasonable benefit/risk
ratio applicable to any medical treatment. In certain embodiments,
the term refers to that amount necessary or sufficient to
eliminate, reduce or maintain a target of a particular therapeutic
regimen. The effective amount may vary depending on such factors as
the disease or condition being treated, the particular targeted
constructs being administered, the size of the subject or the
severity of the disease or condition. One of ordinary skill in the
art may empirically determine the effective amount of a particular
compound without necessitating undue experimentation. In certain
embodiments, a therapeutically effective amount of a therapeutic
agent for in vivo use will likely depend on a number of factors,
including: the rate of release of an agent from a polymer matrix,
which will depend in part on the chemical and physical
characteristics of the polymer; the identity of the agent; the mode
and method of administration; and any other materials incorporated
in the polymer matrix in addition to the agent.
[0061] The term "ED50" refer to the dose of a drug, which produces
50% of its maximum response or effect, or alternatively, the dose,
which produces a pre-determined response in 50% of test subjects or
preparations. The term "LD50" refers to the dose of a drug, which
is lethal in 50% of test subjects. The term "therapeutic index"
refers to the therapeutic index of a drug, defined as
LD50/ED50.
[0062] The term "substituted," as used herein, means that any one
or more hydrogens on the designated atom is replaced with a
selection from the indicated group, provided that the designated
atom's normal valency is not exceeded, and that the substitution
results in a stable compound. When the substituent is keto (i.e.,
.dbd.O), then 2 hydrogens on the atom are replaced. Ring double
bonds, as used herein, are double bonds that are formed between two
adjacent ring atoms (e.g., C.dbd.C, C.dbd.N, or N.dbd.N).
[0063] With respect to any chemical compounds, the present
application is intended to include all isotopes of atoms occurring
in the present compounds. Isotopes include those atoms having the
same atomic number but different mass numbers. By way of general
example and without limitation, isotopes of hydrogen include
tritium and deuterium, and isotopes of carbon include C-13 and
C-14.
[0064] When a bond to a substituent is shown to cross a bond
connecting two atoms in a ring, then such substituent can be bonded
to any atom in the ring. When a substituent is listed without
indicating the atom via which such substituent is bonded to the
rest of the compound of a given formula, then such substituent can
be bonded via any atom in such substituent. Combinations of
substituents and/or variables are permissible, but only if such
combinations result in stable compounds.
[0065] When an atom or a chemical moiety is followed by a
subscripted numeric range (e.g., C.sub.1-6), the invention is meant
to encompass each number within the range as well as all
intermediate ranges. For example, "C.sub.1-6 alkyl" is meant to
include alkyl groups with 1, 2, 3, 4, 5, 6, 1-6, 1-5, 1-4, 1-3,
1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5, and 5-6
carbons.
[0066] As used herein, "alkyl" is intended to include both branched
(e.g., isopropyl, tert-butyl, isobutyl), straight-chain e.g.,
methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl,
decyl), and cycloalkyl (e.g., alicyclic) groups (e.g., cyclopropyl,
cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkyl
substituted cycloalkyl groups, and cycloalkyl substituted alkyl
groups. Such aliphatic hydrocarbon groups have a specified number
of carbon atoms. For example, C.sub.1-6 alkyl is intended to
include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5, and C.sub.6
alkyl groups. As used herein, "lower alkyl" refers to alkyl groups
having from 1 to 6 carbon atoms in the backbone of the carbon
chain. "Alkyl" further includes alkyl groups that have oxygen,
nitrogen, sulfur or phosphorous atoms replacing one or more
hydrocarbon backbone carbon atoms. In certain embodiments, a
straight chain or branched chain alkyl has six or fewer carbon
atoms in its backbone (e.g., C.sub.1-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain), for example four or fewer.
Likewise, certain cycloalkyls have from three to eight carbon atoms
in their ring structure, such as five or six carbons in the ring
structure.
[0067] The term "substituted alkyls" refers to alkyl moieties
having substituents replacing a hydrogen on one or more carbons of
the hydrocarbon backbone. Such substituents can include, for
example, alkyl, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Cycloalkyls can be further substituted, e.g., with the substituents
described above. An "alkylaryl" or an "aralkyl" moiety is an alkyl
substituted with an aryl (e.g., phenylmethyl (benzyl)).
[0068] As used herein, "alkenyl" is intended to include hydrocarbon
chains of either straight or branched configuration having one or
more carbon-carbon double bonds occurring at any stable point along
the chain. For example, C.sub.2-6 alkenyl is intended to include
C.sub.2, C.sub.3, C.sub.4, C.sub.5, and C.sub.6 alkenyl groups.
Examples of alkenyl include, but are not limited to, ethenyl and
propenyl.
[0069] As used herein, "alkynyl" is intended to include hydrocarbon
chains of either straight or branched configuration having one or
more carbon-carbon triple bonds occurring at any stable point along
the chain. For example, C.sub.2-6 alkynyl is intended to include
C.sub.2, C.sub.3, C.sub.4, C.sub.5, and C.sub.6 alkynyl groups.
Examples of alkynyl include, but are not limited to, ethynyl and
propynyl.
[0070] Furthermore, "alkyl", "alkenyl", and "alkynyl" are intended
to include moieties which are diradicals, i.e., having two points
of attachment. A nonlimiting example of such an alkyl moiety that
is a diradical is --CH.sub.2CH.sub.2--, i.e., a C.sub.2 alkyl group
that is covalently bonded via each terminal carbon atom to the
remainder of the molecule.
[0071] "Aryl" includes groups with aromaticity, including 5- and
6-membered "unconjugated", or single-ring, aromatic groups that may
include from zero to four heteroatoms, as well as "conjugated", or
multicyclic, systems with at least one aromatic ring. Examples of
aryl groups include benzene, phenyl, pyrrole, furan, thiophene,
thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole,
oxazole, isooxazole, pyridine, pyrazine, pyridazine, and
pyrimidine, and the like. Furthermore, the term "aryl" includes
multicyclic aryl groups, e.g., tricyclic, bicyclic, e.g.,
naphthalene, benzoxazole, benzodioxazole, benzothiazole,
benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline,
isoquinoline, napthridine, indole, benzofuran, purine, benzofuran,
deazapurine, or indolizine. Those aryl groups having heteroatoms in
the ring structure may also be referred to as "aryl heterocycles",
"heterocycles," "heteroaryls" or "heteroaromatics". The aromatic
ring can be substituted at one or more ring positions with such
substituents as described above, as for example, halogen, hydroxyl,
alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl,
aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl,
arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkylamino, dialkylamino,
arylamino, diaryl amino, and alkylaryl amino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety. Aryl groups can also be fused or
bridged with alicyclic or heterocyclic rings, which are not
aromatic so as to form a multicyclic system (e.g., tetralin,
methylenedioxyphenyl).
[0072] The terms "heterocyclyl" or "heterocyclic group" include
closed ring structures, e.g., 3- to 10-, or 4- to 7-membered rings,
which include one or more heteroatoms. "Heteroatom" includes atoms
of any element other than carbon or hydrogen. Examples of
heteroatoms include nitrogen, oxygen, sulfur and phosphorus.
[0073] Heterocyclyl groups can be saturated or unsaturated and
include pyrrolidine, oxolane, thiolane, piperidine, piperazine,
morpholine, lactones, lactams such as azetidinones and
pyrrolidinones, sultams, and sultones. Heterocyclic groups such as
pyrrole and furan can have aromatic character. They include fused
ring structures such as quinoline and isoquinoline. Other examples
of heterocyclic groups include pyridine and purine. The
heterocyclic ring can be substituted at one or more positions with
such substituents as described above, as for example, halogen,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkyl amino, dialkylamino,
arylamino, diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, or an aromatic or
heteroaromatic moiety. Heterocyclic groups can also be substituted
at one or more constituent atoms with, for example, a lower alkyl,
a lower alkenyl, a lower alkoxy, a lower alkylthio, a lower
alkylamino, a lower alkylcarboxyl, a nitro, a hydroxyl, --CF.sub.3,
or --CN, or the like.
[0074] As used herein, "halo" or "halogen" refers to fluoro,
chloro, bromo, and iodo. "Counterion" is used to represent a small,
negatively charged species such as fluoride, chloride, bromide,
iodide, hydroxide, acetate, and sulfate.
[0075] "Stable compound" and "stable structure" are meant to
indicate a compound that is sufficiently robust to survive
isolation, and as appropriate, purification from a reaction
mixture, and formulation into an efficacious therapeutic agent.
[0076] "Free compound" is used herein to describe a compound in the
unbound state.
[0077] "Extinction coefficient" is a constant used in the
Beer-Lambert Law which relates the concentration of the substance
being measured (in moles) to the absorbance of the substance in
solution (how well the substance in solution blocks light beamed
through it from getting out on the other side). It is an indicator
of how much light a compound absorbs at a particular
wavelength.
[0078] In the specification, the singular forms also include the
plural, unless the context clearly dictates otherwise. Throughout
the description, where compositions are described as having,
including, or comprising, specific components, it is contemplated
that compositions also consist essentially of, or consist of, the
recited components. Similarly, where methods or processes are
described as having, including, or comprising specific process
steps, the processes also consist essentially of, or consist of,
the recited processing steps. Further, it should be understood that
the order of steps or order for performing certain actions is
immaterial so long as the invention remains operable. Moreover, two
or more steps or actions can be conducted simultaneously.
[0079] "Small molecule" refers to a molecule, which has a molecular
weight of less than about 2000 amu, or less than about 1000 amu,
and even less than about 500 amu.
[0080] All percentages and ratios used herein, unless otherwise
indicated, are by weight.
[0081] The term "retina" refers to a region of the central nervous
system with approximately 150 million neurons. It is located at the
back of the eye where it rests upon a specialized epithelial tissue
called retinal pigment epithelium or RPE. The retina initiates the
first stage of visual processing by transducing visual stimuli in
specialized neurons called "photoreceptors". Their synaptic outputs
are processed by elaborate neural networks in the retina and then
transmitted to the brain. The retina has evolved two specialized
classes of photoreceptors to operate under a wide range of light
conditions. "Rod" photoreceptors transduce visual images under low
light conditions and mediate achromatic vision. "Cone"
photoreceptors transduce visual images in dim to bright light
conditions and mediate both color vision and high acuity
vision.
[0082] Every photoreceptor is compartmentalized into two regions
called the "outer" and "inner" segment. The inner segment is the
neuronal cell body containing the cell nucleus. The inner segment
survives for a lifetime in the absence of retinal disease. The
outer segment is the region where the light sensitive visual
pigment molecules are concentrated in a dense array of stacked
membrane structures. Part of the outer segment is routinely shed
and regrown in a diurnal process called outer segment renewal. Shed
outer segments are ingested and metabolized by RPE cells.
[0083] The term "macula" refers to the central region of the
retina, which contains the fovea where visual images are processed
by long slender cones in high spatial detail ("visual acuity").
"Macular degeneration" is a form of retinal neurodegeneration,
which attacks the macula and destroys high acuity vision in the
center of the visual field. AMD can be in a "dry form"
characterized by residual lysosomal granules called lipofuscin in
RPE cells, and by extracellular deposits called "drusen". Drusen
contain cellular waste products excreted by RPE cells. "Lipofuscin"
and drusen can be detected clinically by ophthalmologists and
quantified using fluorescence techniques. They can be the first
clinical signs of macular degeneration.
[0084] Lipfuscin contains aggregations of A2E. Lipofuscin
accumulates in RPE cells and poisons them by multiple known
mechanisms. As RPE cells become poisoned, their biochemical
activities decline and photoreceptors begin to degenerate.
Extracellular drusen may further compromise RPE cells by
interfering with their supply of vascular nutrients. Drusen also
trigger inflammatory processes, which leads to choroidal
neovascular invasions of the macula in one patient in ten who
progresses to wet form AMD. Both the dry form and wet form progress
to blindness.
[0085] The term "ERG" is an acronym for electroretinogram, which is
the measurement of the electric field potential emitted by retinal
neurons during their response to an experimentally defined light
stimulus. ERG is a non-invasive measurement, which can be performed
on either living subjects (human or animal) or a hemisected eye in
solution that has been removed surgically from a living animal.
[0086] The term "RAL" means retinaldehyde. "Free RAL" is defined as
RAL that is not bound to a visual cycle protein. The terms
"trans-RAL" and "all-trans-RAL" are used interchangeably and mean
all-trans-retinaldehyde.
[0087] An embodiment of the application relates to compounds and
methods of treating an ocular disorder in a subject associated with
aberrant all-trans-retinal clearance in the retina. The ocular
disorder can include, for example, retinal disorders, such as
macular degeneration, including age-related macular degeneration,
geographic atrophy (GA), Stargardt disease, and retinitis
pigmentosa. FIGS. 1 and 2 show the retinoid flow in the visual
cycle including condensation of all-trans-RAL, and all-trans-RAL
clearance. After 11-cis-retinal binds to opsin from rhodopsin, the
resulting visual chromophore 11-cis-retinylidene is photoisomerized
to all-trans-retinylidene, the precursor or all-trans-RAL that is
later released. Most of the all-trans-RAL dissociates from opsin
into the cytoplasm before it is reduced to all-trans-retinol by
RDHs including RDH8. The fraction of all-trans-RAL that dissociates
into disc lumens is transported by ABCA4 before is it is reduced.
Thus, condensation products can be generated both within the disc
lumens and the cytoplasm before it is reduced.
[0088] It was found that all-trans-RAL that has escaped
sequestering by opsins in photoreceptor outer segments of the
retina is toxic to retina cells and that aberrant all-trans-RAL
clearance from the inner disc membrane to the outer disc membrane
can cause retinal degeneration. The mechanism of all-trans-RAL
toxicity can include plasma membrane permeability and mitochondrial
poisoning that leads to caspase activation and mitochondrial
associated cell death.
[0089] In accordance with an embodiment of the application,
compounds used to treat an ocular disorder associated with aberrant
all-trans-RAL clearance can include primary amines (i.e., primary
amine compounds) that form reversible Schiff-bases with free
all-trans-RAL, which has escaped sequestering in photoreceptor
outer segments of the retina without adversely affecting normal
retinoid cycle. Formation of a reversible Schiff base between RAL
and the primary amine compounds described herein can control or
modulate all-trans-RAL levels in the retina and prevent retina
degeneration. The stability of the Schiff-bases formed between the
primary amine compounds and the free RAL under physiologic
conditions of the retina can be used to determine the efficacy of
these compounds in treating the ocular disorder. The stability of
the Schiff-bases formed from the primary amine compounds should be
such that the level of free RAL in the retina is reduced to a level
that is effective to mitigate retinal degeneration but not impair
the normal retinoid cycle.
[0090] In an embodiment of the application, the primary amine
compounds that can form stable Schiff-bases with all-trans-RAL
under physiological conditions of the retina and that can inhibit
retinal degeneration upon administration to a subject can be
selected using an in vitro assay that measures the ability of a
primary amine compound to form a Schiff base with retinal under
physiological condition of the retina and in vivo assays that
measure, respectively, 11-cis-retinal formation and the optical
coherence tomography score of retinas of Rdh8.sup.-/-Abca4.sup.-/-
mice. Primary amine compounds that form a Schiff-base with
all-trans-RAL or its metabolite under physiologic conditions of the
retina and that when administered to a Rdh8.sup.-/-Abca4.sup.-/-
mouse increase the optical coherence tomography score of the mouse
to at least about 2.5 and increase 11-cis-retinal amount at least
about 30% in comparison to untreated control animal are effective
in treating retinal degeneration in a subject associated with
aberrant all-trans-RAL clearance. Primary amines compounds that do
not form a form a Schiff-base with all-trans-RAL or its metabolite
under physiologic conditions of the retina or which when
administered to a Rdh8.sup.-/- Abca4.sup.-/- mouse do not increase
the optical coherence tomography score of the mouse to at least
about 2.5 and increase 11-cis-retinal amount at least about 30% in
comparison to untreated control animal, were found to be
ineffective in treating retinal degeneration in a subject
associated with aberrant all-trans-RAL clearance. Additionally,
therapeutic efficacy of the primary amine compounds of the
application can be determined using an in vitro assay that measures
the ability of a primary amine compound to improve viability of RPE
cells treated with retinal.
[0091] In some embodiments, the primary amine compound can include
the structural formula (I):
##STR00003##
[0092] wherein R.sub.1 is an aliphatic and/or aromatic
compound.
[0093] Primary amine compounds having formula I that are used to
treat retinal degeneration in accordance with an embodiment of the
application can upon administration to the subject form a
reversible Schiff-base with the all-trans-RAL without adversely
affecting normal retinoid cycle performance and when administered
to a Rdh8.sup.-/-Abca4.sup.-/- mouse increase the optical coherence
tomography score of the mouse to at least about 2.5 and increase
11-cis-retinal amount at least about 30% in comparison to untreated
control animal. Primary amine compounds in accordance with the
application, however, do not include and are not a local
anesthetic, which includes an aromatic amine that demonstrates
sodium channel blockade when administered to the subject.
[0094] Advantageously, the primary amine compounds in accordance
with the application do not inhibit RPE65 enzymatic activity or any
other proteins involved in retinoid metabolism in the eye of the
subject. The primary amine compounds can reduce the formation of
A2E and/or retinal dimer in the subject's retina, promote
11-cis-retinal production in the subject, and does not cause night
blindness.
[0095] In some embodiments, primary compounds having formula I that
upon administration to a subject form a reversible Schiff-base with
the all-trans-RAL without adversely affecting normal retinoid cycle
performance and that when administered to a Rdh8.sup.-/-
Abca4.sup.-/- mouse increase the optical coherence tomography score
of the mouse to at least about 2.5 and increase 11-cis-retinal
amount at least about 30% in comparison to untreated control animal
can be selected using the methods described in the Examples from
known primary amine compounds.
[0096] In an embodiment of the application, the primary amine
compounds can include known primary amine compounds having the
following structural formulas:
##STR00004##
[0097] wherein R.sub.2 is hydrogen or (C.sub.1-C.sub.6) straight
chain or branched unsubstituted or substituted alkyl;
[0098] R.sub.3 is straight or branched unsubstituted or substituted
alkyl of from 1 to 8 carbon atoms, straight or branched alkenyl of
from 2 to 8 carbon atoms, cycloalkyl of from 3 to 7 carbon atoms,
alkoxy of from 1 to 6 carbon atoms, -alkylcycloalkyl, -alkylalkoxy,
-alkyl, OH, -alkylphenyl, -alkylphenoxy, -phenyl or substituted
phenyl;
[0099] R.sub.4 is hydrogen or (C.sub.1-C.sub.6) straight chain or
branched unsubstituted or substituted alkyl, or carboxyl;
[0100] Ar is phenyl which is unsubstituted or substituted with 1-5
of R.sub.7, wherein R.sub.7 is independently selected from the
group consisting of: [0101] (1) halogen, [0102] (2) C.sub.1-6
alkyl, which is linear or branched and is unsubstituted or
substituted with 1-5 halogens, [0103] (3) OC.sub.1-6 alkyl, which
is linear or branched and is unsubstituted or substituted with 1-5
halogens, and [0104] (4) CN;
[0105] X.sub.1 is selected from the group consisting of: [0106] (1)
N, and [0107] (2) CR.sub.6;
[0108] R.sub.5 and R.sub.6 are independently selected from the
group consisting of: [0109] (1) hydrogen, [0110] (2) CN, [0111] (3)
C.sub.1-10 alkyl, which is linear or branched and which is
unsubstituted or substituted with 1-5 halogens or phenyl, which is
unsubstituted or substituted with 1-5 substituents independently
selected from halogen, CN, OH, R.sub.8, OR.sub.8,
NHSO.sub.2R.sub.8, SO.sub.2R.sub.8, CO.sub.2H, and
CO.sub.2C.sub.1-6 alkyl, wherein the CO.sub.2C.sub.1-6 alkyl is
linear or branched, [0112] (4) phenyl which is unsubstituted or
substituted with 1-5 substituents independently selected from
halogen, CN, OH, R.sub.8, OR.sub.8, NHSO.sub.2R.sub.8,
SO.sub.2R.sub.8, CO.sub.2H, and CO.sub.2C.sub.1-6 alkyl, wherein
the CO.sub.2C.sub.1-6 alkyl is linear or branched, and [0113] (5) a
5- or 6-membered heterocycle which may be saturated or unsaturated
comprising 1-4 heteroatoms independently selected from N, S and O,
the heterocycle being unsubstituted or substituted with 1-3
substituents independently selected from oxo, OH, halogen,
C.sub.1-6 alkyl, and OC.sub.1-6 alkyl, wherein the C.sub.1-6 alkyl
and OC.sub.1-6 alkyl are linear or branched and optionally
substituted with 1-5 halogens;
[0114] R.sub.8 is C.sub.1-6 alkyl, which is linear or branched and
which is unsubstituted or substituted with 1-5 groups independently
selected from halogen, CO.sub.2H, and CO.sub.2C.sub.1-6 alkyl,
wherein the CO.sub.2C.sub.1-6 alkyl is linear or branched;
[0115] R.sub.9 and R.sub.10 may be the same or different and are
hydrogen, straight or branched alkyl of from one to six carbon
atoms, lower alkylaryl, lower alkenyl, phenyl, CF.sub.3, hydroxy,
lower alkoxy, lower alkylthio, lower alkylsulphonyl, CF.sub.3O, at
the six position halogen, nitro, carboxy, lower alkoxycarbonyl,
NR.sub.11R.sub.12CO, NR.sub.11R.sub.12, R.sub.11CONR.sub.12, CN,
NR.sub.11R.sub.12SO.sub.2, wherein R.sub.11 and R.sub.12 may be the
same or different and are hydrogen, lower alkyl, or aryl; R.sub.9
and R.sub.10 may together form a carbocyclic or methylenedioxy
ring;
[0116] R.sub.14 is cyano, cyanomethyl, methoxymethyl, or
ethoxymethyl;
[0117] X.sub.2 is O, N(H), or S, het is a 5 or 6-membered
heterocycle, n is 0, 1, 2, or 3, and each D is an unbranched lower
alkyl group;
[0118] U is a substituent selected from halogen atom; cyano; lower
alkyl wherein one or more hydrogen atoms on the lower alkyl group
are optionally substituted by groups selected from a halogen atom,
hydroxyl, carbamoyl, amino, aryl, and a monocyclic or bicyclic
heterocyclic group containing one or more hetero-atoms selected
from nitrogen, oxygen, and sulfur atoms; lower alkylthio wherein
one or more hydrogen atoms on the alkyl group are optionally
substituted by groups selected from a halogen atom, hydroxyl,
carbamoyl, amino, and aryl; lower alkylsulfonyl wherein one or more
hydrogen atoms on the alkyl group are optionally substituted by
groups selected from a halogen atom, hydroxyl, carbamoyl, amino,
and aryl; hydroxyl; lower alkoxy; formyl; lower alkylcarbonyl;
arylcarbonyl; carboxyl; lower alkoxycarbonyl; carbamoyl; N-lower
alkylcarbamoyl; N,N-di-lower alkylaminocarbonyl; amino; N-lower
alkylamino; N,N-di-lower alkylamino; formylamino; lower
alkylcarbonylamino; aminosulfonylamino; (N-lower
alkylamino)sulfonylamino; (N,N-di-lower alkylamino)sulfonylamino;
aryl, optionally substituted by groups selected from a halogen
atom, hydroxyl, carbamoyl, aryl and amino; and a monocyclic or
bicyclic heterocyclic group containing one or more hetero-atoms
selected from nitrogen, oxygen, and sulfur atoms;
[0119] Q, T, and V are each, independently, N, S, O CU or CH;
[0120] W, X, Y, and Z are each, independently, N, S, O CU or CH,
such that at least one of W, X, Y, and Z is N;
[0121] A is
##STR00005##
[0122] D is unbranched lower alkyl;
[0123] R.sub.15 and R.sub.16 are each independently substituted or
unsubstituted C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5, C.sub.6,
C.sub.7, or C.sub.8, straight chain alkyl, or substituted or
unsubstituted C.sub.3, C.sub.4, C.sub.5, C.sub.6, C.sub.7, or
C.sub.8, branched chain alkyl;
[0124] L is a single bond or CH.sub.2;
[0125] m is 0, 1, or 2;
[0126] n is 0, 1, 2, 3, or 4;
[0127] Y.sub.1 is --(CH.sub.2).sub.2--, --(CH.sub.2).sub.3--,
--CH.sub.2CH(CH.sub.3)-- or --CH.sub.2C(CH.sub.3).sub.2--;
[0128] R.sub.17 is aryl or heteroaryl;
[0129] R.sub.18 and R.sub.19 are each independently C.sub.1-C.sub.4
alkyl or 2-methoxyethyl;
[0130] R.sub.20 is hydrogen, C.sub.1-C.sub.4 alkyl,
2-(C.sub.1-C.sub.4 alkoxy)ethyl, cyclopropylmethyl, benzyl, or
--(CH.sub.2).sub.m1COR.sub.21 where m1 is 1, 2 or 3 and R.sup.21 is
hydroxy, C.sub.1-C.sub.4 alkoxy or --NR.sub.22 where R.sub.22
hydrogen or C.sub.1-C.sub.4 alkyl;
[0131] R.sub.23 and R.sub.24 can be the same or different and are
hydrogen, methyl, or ethyl
[0132] as well as pharmaceutically acceptable salts thereof.
[0133] In other embodiments, the primary amine compound can be
selected from the group consisting of:
##STR00006## ##STR00007##
[0134] and pharmaceutically acceptable salts thereof.
[0135] In a still further embodiment, the primary amine compound
can be selected from the group consisting of:
##STR00008##
[0136] and pharmaceutically acceptable salts thereof.
[0137] In another embodiment, the primary compound can have the
following structural formula:
##STR00009##
[0138] wherein Ar is phenyl which is unsubstituted or substituted
with 1-5 substitutents which are independently selected from the
group consisting of: (1) fluoro, (2) bromo, and (3) CF.sub.3,
R.sub.5 is selected from the group consisting of: (1) hydrogen, and
(2) C.sub.1-6 alkyl, which is linear or branched and which is
unsubstituted or substituted with phenyl or 1-5 fluoro.
[0139] In yet another embodiment, it is more preferred that Ar is
selected from the group consisting of: (1) phenyl, (2)
2-fluorophenyl, (3) 3,4-difluorophenyl, (4) 2,5-difluorophenyl, (5)
2,4,5-trifluorophenyl, (6) 2-fluoro-4-(trifluoromethyl)phenyl, and
(7) 4-bromo-2,5-difluorophenyl and R5 is selected from the group
consisting of: (1) hydrogen, (2) methyl, (3) ethyl, (4) CF.sub.3,
(5) CH.sub.2CF.sub.3, (5) CF.sub.2CF.sub.3 (6) phenyl, and (7)
benzyl.
[0140] In another embodiment, the primary amine compound can be a
monobasic dihydrogenphosphate salt of
4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8-
H)-yl]-1-(2,4,5-trifluorophenyl) butan-2-amine of the following
structural formula:
##STR00010##
[0141] or a crystalline hydrate thereof. The crystalline hydrate
can be a crystalline monohydrate of the dihydrogenphosphate
salt.
[0142] The dihydrogenphosphate salt shown above has a center of
asymmetry at the stereogenic carbon atom indicated with an * and
can thus occur as a racemate, racemic mixture, and single
enantiomers, with all isomeric forms being included in the present
invention. The separate enantiomers, substantially free of the
other, are included within the scope of the invention, as well as
mixtures of the two enantiomers. Monobasic dihydrogenphosphate salt
of
4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8-
H)-yl]-1-(2,4,5-trifluorophenyl) butan-2-amine.
[0143] In a further embodiment, the primary amine compound can be a
dihydrogenphosphate salt of
(2R)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazi-
n-7(8H)-yl]-1-(2,4,5-triflorophenyl) butan-2-amine of the following
structural formula:
##STR00011##
[0144] or a crystalline hydrate thereof.
[0145] In yet another embodiment, the primary amine compound can be
a dihydrogenphosphate salt of
(2S)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazi-
-n-7(8H)-yl]-1-(2,4,5-trifluorophenyl) butan-2-amine of the
following structural formula:
##STR00012##
[0146] or a crystalline hydrate thereof.
[0147] In some embodiments, the primary amine compound is a
compound having the following structural formula:
##STR00013##
[0148] wherein R.sub.2 is hydrogen or (C.sub.1-C.sub.6) straight
chain or branched unsubstituted or substituted alkyl;
[0149] R.sub.3 is straight or branched unsubstituted or substituted
alkyl of from 1 to 8 carbon atoms, straight or branched alkenyl of
from 2 to 8 carbon atoms, cycloalkyl of from 3 to 7 carbon atoms,
alkoxy of from 1 to 6 carbon atoms, -alkylcycloalkyl, -alkylalkoxy,
-alkyl, OH, -alkylphenyl, -alkylphenoxy, -phenyl or substituted
phenyl;
[0150] R.sub.4 is hydrogen or (C.sub.1-C.sub.6) straight chain or
branched unsubstituted or substituted alkyl, or carboxyl;
[0151] as well as pharmaceutically acceptable salts thereof.
[0152] In other embodiments, the primary amine compound is a
compound having the following structural formula:
##STR00014##
[0153] wherein R.sub.2 is hydrogen, straight or branched alkyl of
from 1 to 6 carbon atoms or phenyl;
[0154] R.sub.3 is straight or branched alkyl of from 1 to 8 carbon
atoms, straight or branched alkenyl of from 2 to 8 carbon atoms,
cycloalkyl of from 3 to 7 carbon atoms, alkoxy of from 1 to 6
carbon atoms, -alkylcycloalkyl, -alkylalkoxy, -alkyl
OH-alkylphenyl, -alkylphenoxy, -phenyl or substituted phenyl;
and
[0155] R.sub.4 is hydrogen, and R.sub.2 is straight or branched
alkyl of from 1 to 6 carbon atoms or phenyl when R.sub.3 is methyl,
or a pharmaceutically acceptable salt thereof.
[0156] In other embodiments, the primary amine compound can have
the following structural formula:
##STR00015##
[0157] wherein R.sub.2 is methyl, R.sub.3 is an alkyl, and R.sub.4
is a hydrogen, or a pharmaceutically acceptable salt thereof.
[0158] Specific examples of compounds of above noted formulas are
selected from: 3-Aminomethyl-5-methylhexanoic acid;
3-Aminomethyl-5-methylheptanoic acid;
3-Aminomethyl-5-methyl-octanoic acid;
3-Aminomethyl-5-methyl-nonanoic acid;
3-Aminomethyl-5-methyl-decanoic acid;
3-Aminomethyl-5-methyl-undecanoic acid;
3-Aminomethyl-5-methyl-dodecanoic acid;
3-Aminomethyl-5-methyl-tridecanoic acid;
3-Aminomethyl-5-cyclopropyl-hexanoic acid;
3-Aminomethyl-5-cyclobutyl-hexanoic acid;
3-Aminomethyl-5-cyclopentyl-hexanoic acid;
3-Aminomethyl-5-cyclohexyl-hexanoic acid;
3-Aminomethyl-5-trifluoromethyl-hexanoic acid;
3-Aminomethyl-5-phenyl-hexanoic acid;
3-Aminomethyl-5-(2-chlorophenyl)-hexanoic acid;
3-Aminomethyl-5-(3-chlorophenyl)-hexanoic acid;
3-Aminomethyl-5-(4-chlorophenyl)-hexanoic acid;
3-Aminomethyl-5-(2-methoxyphenyl)-hexanoic acid;
3-Aminomethyl-5-(3-methoxyphenyl)-hexanoic acid;
3-Aminomethyl-5-(4-methoxyphenyl)-hexanoic acid;
3-Aminomethyl-5-(phenylmethyl)-hexanoic acid;
(S)-3-(Aminomethyl)-5-methylhexanoic acid;
(R)-3-(Aminomethyl)-5-methylhexanoic acid;
(3R,4S)-3-Aminomethyl-4,5-dimethyl-hexanoic acid;
3-Aminomethyl-4,5-dimethyl-hexanoic acid;
(3R,4S)-3-Aminomethyl-4,5-dimethyl-hexanoic acid MP;
(3S,4S)-3-Aminomethyl-4,5-dimethyl-hexanoic acid;
(3R,4R)-3-Aminomethyl-4,5-dimethyl-hexanoic acid MP;
3-Aminomethyl-4-isopropyl-hexanoic acid;
3-Aminomethyl-4-isopropyl-heptanoic acid;
3-Aminomethyl-4-isopropyl-octanoic acid;
3-Aminomethyl-4-isopropyl-nonanoic acid;
3-Aminomethyl-4-isopropyl-decanoic acid;
3-Aminomethyl-4-phenyl-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-ethoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-propoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-isopropoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-tert-butoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-fluoromethoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-fluoro-ethoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3,3,3-trifluoro-propoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-phenoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-chloro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-chloro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-chloro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-fluoro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-fluoro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-fluoro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-methoxy-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-methoxy-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-methoxy-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(4-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(3-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-(2-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-hydroxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-methoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-ethoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-propoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-isopropoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-tert-butoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-fluoromethoxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-fluoro-ethoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-(3,3,3-trifluoro-propoxy)-hexanoic
acid; (3S,5S)-3-Aminomethyl-5-methyl-6-phenoxy-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-chloro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-chloro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-chloro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-fluoro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-fluoro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-fluoro-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-methoxy-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-methoxy-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-methoxy-phenoxy)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl
6-(4-trifluoromethyl-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl
6-(3-trifluoromethyl-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl
6-(2-trifluoromethyl-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl 6-(4-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl 6-(3-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl 6-(2-nitro-phenoxy)-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-benzyloxy-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-7-hydroxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-methoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-ethoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-propoxy-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-isopropoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-tert-butoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-fluoromethoxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(2-fluoro-ethoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(3,3,3-trifluoro-propoxy)-heptanoi-c
acid; (3S,5S)-3-Aminomethyl-7-benzyloxy-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-phenoxy-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(4-chloro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(3-chloro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(2-chloro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(4-fluoro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(3-fluoro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(2-fluoro-phenoxy)-5-methyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-7-(4-methoxy-phenoxy)-5-methyl-heptanoic
acid;
(3S,5S)-3-Aminomethyl-7-(3-methoxy-phenoxy)-5-methyl-heptanoic
acid;
(3S,5S)-3-Aminomethyl-7-(2-methoxy-phenoxy)-5-methyl-heptanoic
acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(4-trifluoromethyl-phenoxy)-heptan-oic
acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(3-trifluoromethyl-phenoxy)-heptan-
-oic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(2-trifluoromethyl-phenoxy)-he-
ptan-oic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(4-nitro-phenoxy)-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(3-nitro-phenoxy)-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-7-(2-nitro-phenoxy)-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-phenyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-chloro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-chloro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-chloro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-methoxy-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-methoxy-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-methoxy-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(4-fluoro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(3-fluoro-phenyl)-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-6-(2-fluoro-phenyl)-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-7-phenyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(4-chloro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(3-chloro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(2-chloro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(4-methoxy-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(3-methoxy-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(2-methoxy-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(4-fluoro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(3-fluoro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-(2-fluoro-phenyl)-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-oct-7-enoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-non-8-enoic acid;
(E)-(3S,5S)-3-Aminomethyl-5-methyl-oct-6-enoic acid;
(Z)-(3S,5S)-3-Aminomethyl-5-methyl-oct-6-enoic acid;
(Z)-(3S,5S)-3-Aminomethyl-5-methyl-non-6-enoic acid;
(E)-(3S,5S)-3-Aminomethyl-5-methyl-non-6-enoic acid;
(E)-(3S,5R)-3-Aminomethyl-5-methyl-non-7-enoic acid;
(Z)-(3S,5R)-3-Aminomethyl-5-methyl-non-7-enoic acid;
(Z)-(3S,5R)-3-Aminomethyl-5-methyl-dec-7-enoic acid;
(E)-(3S,5R)-3-Aminomethyl-5-methyl-undec-7-enoic acid;
(3S,5S)-3-Aminomethyl-5,6,6-trimethyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5,6-dimethyl-heptanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclopropyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclobutyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclopentyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-5-cyclohexyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-nonanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-decanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-undecanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-dodecanoic acid;
(3S,5R)-3-Aminomethyl-5,9-dimethyl-decanoic acid;
(3S,5R)-3-Aminomethyl-5,7-dimethyl-octanoic acid;
(3S,5R)-3-Aminomethyl-5,8-dimethyl-nonanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclopropyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclobutyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclopentyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-6-cyclohexyl-5-methyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclopropyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclobutyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclopentyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-7-cyclohexyl-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclopropyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclobutyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclopentyl-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-8-cyclohexyl-5-methyl-octanoic acid;
(3S,5S)-3-Aminomethyl-6-fluoro-5-methyl-hexanoic acid;
(3S,5S)-3-Aminomethyl-7-fluoro-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-8-fluoro-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-9-fluoro-5-methyl-nonanoic acid;
(3S,5S)-3-Aminomethyl-7,7,7-trifluoro-5-methyl-heptanoic acid;
(3S,5R)-3-Aminomethyl-8,8,8-trifluoro-5-methyl-octanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-8-phenyl-octanoic acid;
(3S,5S)-3-Aminomethyl-5-methyl-6-phenyl-hexanoic acid;
(3S,5R)-3-Aminomethyl-5-methyl-7-phenyl-heptanoic acid; and
pharmaceutically acceptable salts thereof. Methods of synthesizing
the above noted compounds are described in PCT Patent Application
No. WO 00/76958, which is incorporated herein by reference in its
entirety.
[0159] In other embodiments, the primary amine compound can
comprise at least one of (S)-3-(Aminomethyl)-5-methylhexanoic acid
or (R)-3-(Aminomethyl)-5-methylhexanoic acid. In still other
embodiments, the primary amine compound can include a mixture of
(S)-3-(Aminomethyl)-5-methylhexanoic acid and
(R)-3-(Aminomethyl)-5-methylhexanoic acid. For example, the primary
amine compound can comprise a racemic mixture of
(S)-3-(Aminomethyl)-5-methylhexanoic acid and
(R)-3-(Aminomethyl)-5-methylhexanoic acid. In other examples, the
primary amine compound can comprise a mixture of: less than about
50% by weight (S)-3-(Aminomethyl)-5-methylhexanoic acid and greater
than about 50% by weight (R)-3-(Aminomethyl)-5-methylhexanoic acid,
less than about 25% by weight (S)-3-(Aminomethyl)-5-methylhexanoic
acid and greater than about 75% by weight
(R)-3-(Aminomethyl)-5-methylhexanoic acid, less than about 10% by
weight (S)-3-(Aminomethyl)-5-methylhexanoic acid and greater than
about 90% by weight (R)-3-(Aminomethyl)-5-methylhexanoic acid, less
than about 1% by weight (S)-3-(Aminomethyl)-5-methylhexanoic acid
and greater than about 99% by weight
(R)-3-(Aminomethyl)-5-methylhexanoic acid, greater than about 50%
by weight (S)-3-(Aminomethyl)-5-methylhexanoic acid and less than
about 50% by weight (R)-3-(Aminomethyl)-5-methylhexanoic acid,
greater than about 75% by weight
(S)-3-(Aminomethyl)-5-methylhexanoic acid and less than about 25%
by weight (R)-3-(Aminomethyl)-5-methylhexanoic acid, greater than
about 90% by weight (S)-3-(Aminomethyl)-5-methylhexanoic acid and
less than about 10% by weight (R)-3-(Aminomethyl)-5-methylhexanoic
acid, or greater than about 99% by weight
(S)-3-(Aminomethyl)-5-methylhexanoic acid and less than about 1% by
weight (R)-3-(Aminomethyl)-5-methylhexanoic acid.
[0160] In a still further embodiment, the primary amine compound
can consist essentially of or consist of
(S)-3-(Aminomethyl)-5-methylhexanoic acid. In yet another
embodiment, the primary amine compound can consist essentially of
or consist of (R)-3-(Aminomethyl)-5-methylhexanoic acid.
[0161] In some embodiments, the primary amine compound is a
compound having the following structural formula:
##STR00016##
[0162] wherein R.sub.25 is hydrogen or a lower alky, such as a
(C.sub.1-C.sub.6) straight chain or branched unsubstituted or
substituted alkyl, n is 4, 5, or 6 and pharmaceutically acceptable
salts thereof. Compounds having the above noted structural formula
and methods of forming such compounds are described in U.S. Pat.
No. 4,024,175, which is incorporated by reference in its
entirety.
[0163] In some embodiments, the primary amine compound is a
compound having the following structural formula:
##STR00017##
[0164] as well as pharmaceutically acceptable salts thereof. A
primary compound having this structural formula is also referred to
as gabapentin and is sold under the trade name Neurontin.
[0165] In another embodiment, the primary amine compound can have
the following structural formula:
##STR00018##
[0166] wherein R.sub.14 is a methoxymethyl or ethoxymethyl groups.
Methods of synthesizing the above noted compounds are described in
U.S. Pat. No. 4,085,225, which is herein incorporated by reference
in its entirety.
[0167] In a further embodiment, the primary amine compound can have
the following structural formula:
##STR00019##
[0168] wherein R.sub.17 is 2-chlorophenyl, 2-fluorophenyl,
2-methoxyphenyl, 3-chlorophenyl, 2-chloro-3-hydroxyphenyl,
2-chloro-6-fluorophenyl, unsubstituted phenyl or
2,3-dichlorophenyl; R.sub.18 is preferably CH.sub.3; R.sub.19 is
C.sub.2H.sub.5; R.sub.20 is H or CH.sub.3; and Y.sub.1 is
(CH.sub.2).sub.2 or CH.sub.2CH(CH.sub.3). Methods of synthesizing
the above noted compounds are described in U.S. Pat. No. 4,572,909,
which is herein incorporated by reference in its entirety.
[0169] In a further embodiment, the primary amine compound can have
the following structural formula:
##STR00020##
[0170] wherein R.sub.9 and R.sub.10 are hydrogen, straight or
branched alkyl of from one to six carbon atoms, lower alkylaryl,
alkenyl, phenyl, CF.sub.3, lower alkoxy, lower alkylthio, lower
alkylsulphonyl, CF.sub.3O at the six position, halogen, nitro,
NR.sub.11R.sub.12, R.sub.11CONR.sub.11, or CN.
[0171] Examples of compounds having the above noted structure are:
2-aminobenzothiazole, 2-amino-6-methylbenzothiazole,
2-amino-4-methylbenzothiazole,
2-amino-6-trifluoromethylbenzothiazole,
2-amino-4-trifluoromethylbenzothiazole,
2-amino-5-trifluoromethylbenzothiazole,
2-amino-6-trifluoromethoxybenzothiazole,
2-amino-6-ethoxybenzothiazole, 2-amino-6-nitrobenzothiazole,
2-amino-4-methoxybenzothiazole, 2-amino-5-methoxybenzothiazole,
2-amino-4,6-dimethylbenzothiazole, 2-amino-6-bromobenzothiazole,
2-amino-6-chlorobenzothiazole, 2-amino-4-chlorobenzothiazole,
2-amino-6-fluoromethylbenzothiazole, 2-amino-naptho[1,2-d]thiazole,
2-ethylaminobenzothiazole,
2-[[2-(1-methyl-2-pyrrolidinyl)ethyl]amino]-benzothiazole,
2-amino-6-methylsulphonylbenzothiazole,
2-amino-4,6-difluorobenzothiazole,
2-amino-6-methylthiobenzothiazole, 2-benzylaminobenzothiazole, and
pharmaceutically acceptable salts thereof. Methods of synthesizing
the above noted compounds are described in U.S. Pat. No. 4,826,860,
which is herein incorporated by reference in its entirety.
[0172] In some embodiments, the primary amine compound can be
selected using an in vitro assay that measures the ability of the
primary amine compound to improve viability of RPE cells treated
with retinal. By way of example, primary amine compounds
administered to RPE cells treated with retinal that improved the
viability of the RPE cells at least 15% compared to untreated cells
are selected from the group consisting of:
##STR00021## ##STR00022## ##STR00023## ##STR00024## ##STR00025##
##STR00026## ##STR00027## ##STR00028## ##STR00029## ##STR00030##
##STR00031## ##STR00032## ##STR00033## ##STR00034## ##STR00035##
##STR00036## ##STR00037## ##STR00038## ##STR00039## ##STR00040##
##STR00041## ##STR00042## ##STR00043## ##STR00044##
[0173] and pharmaceutically acceptable salts thereof.
[0174] In another example, primary amine compounds administered to
RPE cells treated with retinal that improved the viability of the
RPE cells at least 15% compared to untreated cells are selected
from the group consisting of:
5-amino-2,3-dihydrophthalazine-1,4-dione, 3,4-diethoxyaniline,
1-isopropyl-2-methyl-benzimidazol-5-amine,
N2-(4-dimethylaminophenyl)-1,3-benzothiazole-2,6-diamine,
N-[(3-aminophenyl)methyl]-6-methoxy-chroman-4-amine,
1-[[4-(aminomethyl)phenyl]methyl]hexahydropyrimidin-2-one,
1-(2,4-diphenylpyrimidin-5-yl)ethanamine,
3-(5-aminopentyl)-1-[(E)-(5-nitro-2-furyl)methyleneamino]imidazolidine-2,-
4-dione,
2-amino-N-[1-[[1-[(2-amino-1-benzyl-2-oxo-ethyl)carbamoyl]-2-meth-
yl-propyl]carbamoyl]-3-methyl-butyl]-4-methyl-pentanamide,
2-(2-furyl)bicyclo[2.2.1]hept-5-en-3-amine,
5-(3-aminophenyl)furan-2-carboxamidine,
3-(3-aminopropanoyl)-1-[(E)-[5-(4-methoxyphenyl)-2-furyl]methyleneamino]i-
midazolidine-2,4-dione, 4-amino-N-(2-amino-2-oxo-ethyl)benzamide,
4-amino-N-[2-oxo-2-[(2-oxooxazolidin-3-yl)amino]ethyl]benzamide,
(1S,2S,4R)-2-amino-4-isopropenyl-1-methyl-cyclohexanol,
2-amino-4-benzyl-phenol,
(3S,5R,8R,9S,10S,13R,14S)-14-amino-3-hydroxy-10,13-dimethyl-1,2,3,4,5,6,7-
,8,9,11,12,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-17-carboxylic
acid, methyl
(3S,5R,8R,9S,10S,13R,14S)-14-amino-3-[(2S,5R)-5-hydroxy-6-methyl-tetrahyd-
ropyran-2-yl]oxy-10,13-dimethyl-1,2,3,4,5,6,7,8,9,11,12,15,16,17-tetradeca-
hydrocyclopenta[a]phenanthrene-17-carboxylate,
1-[(E)-[5-(4-aminophenyl)-2-furyl]methyleneamino]-3-[4-(4-methylpiperazin-
-1-yl)butyl]imidazolidine-2,4-dione, 4-amino-2-hydroxy-benzoic
acid, fluoranthen-3-amine, phenazine-2,3-diamine,
3-chloro-4-(4-chlorophenoxy)aniline,
4-(6-methyl-1,3-benzothiazol-2-yl)aniline,
3-[5-(1H-benzimidazol-2-yl)-2-furyl]aniline,
N-(2-aminoethyl)-7-tert-butyl-3,3-dimethyl-2H-benzofuran-5-carboxamide,
N'-benzylpropane-1,3-diamine,
5,6-dihydro-2-methyl-4H-pyrrolo[3,2,1-ij]quinoline-1-propanamine,
5-(4-aminophenyl)-2-(o-tolyl)pyrazol-3-amine,
(2,3-dimethyl-1H-indol-5-yl)methanamine,
2,4-dimethyl-6-nitro-aniline, methyl
2-amino-4,5-dimethoxy-benzoate,
2-(5-propyl-1H-indol-3-yl)ethanamine,
2-(7-methoxy-5-nitro-1H-indol-3-yl)ethanamine,
5-amino-2-[(4-carboxyphenyl)carbamoyl]benzoic acid,
5-amino-2-[(3-carboxyphenyl)carbamoyl]benzoic acid,
[2-[2-(3-aminobenzoyl)oxyphenyl]phenyl]3-aminobenzoate,
[4-[1-[4-(4-aminobenzoyl)oxyphenyl]-1-methyl-ethyl]phenyl]4-aminobenzoate-
, 4-amino-N'-(4-chlorobenzoyl)benzohydrazide,
3-(4-aminophenyl)propanoic acid,
2,1,3-benzothiadiazole-4,5-diamine,
1H-benzimidazol-2-ylmethanamine,
2-amino-1-[16-(2-aminoacetyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadec-7-
-yl]ethanone, methyl 6-(2-aminophenyl)-6-oxo-hexanoate,
2-(3-amino-4-ethyl-phenyl)pyridin-3-ol,
(5-amino-6,7-dimethoxy-3-methyl-benzofuran-2-yl)-morpholino-methanone,
(3,5-diaminophenyl)methyl N-butylcarbamate,
(3,5-diaminophenyl)methyl N-(2,4-dimethoxyphenyl)carbamate,
1-(4-aminophenyl)-3-(3,4-difluorophenyl)-1-phenyl-propan-2-one,
N-(2-aminoethyl)-2-[bis(2-hydroxyethyl)amino]acetamide,
(Z)--N-(2-aminoethyl)-3-(1-naphthyl)prop-2-enamide,
N-(2-aminoethyl)naphthalene-1-carboxamide,
(2-amino-5-chloro-phenyl)-phenyl-methanone,
4-(4-bromophenoxy)aniline, 3-aminophenazin-2-ol,
5-amino-N-butyl-2-hydroxy-benzenesulfonamide, ethyl
2-[(2-aminophenyl)carbamothioylamino]acetate,
2-(2-aminophenyl)sulfanyl-4,6-dimethyl-pyridine-3-carbonitrile,
2-amino-1-phenyl-ethanone, 2-(2-methylphenoxy)aniline,
(2-amino-5-chloro-phenyl)-(2-chlorophenyl)methanone,
(1-phenylcyclopentyl)methanamine, tetralin-5-amine,
2-amino-3-(2-hydroxyphenyl)propanoic acid,
3-aminopropane-1-sulfinic acid,
(3R,4R,5R)-2-[(1S,2S)-4,6-diamino-3-[(2R,3R)-3-amino-6-[1-(methylam-
ino)ethyl]tetrahydropyran-2-yl]oxy-2-hydroxy-cyclohexoxy]-5-methyl-4-(meth-
ylamino)tetrahydropyran-3,5-diol, 4-ethoxyaniline,
N-(4-amino-5-chloro-2-hydroxy-phenyl)benzenesulfonamide,
3-amino-N-(3,5-dichloro-2-hydroxy-4-methyl-phenyl)benzamide,
5,6,7,8-tetrahydrophenanthren-2-amine,
2-amino-N-(2-amino-1-benzyl-2-oxo-ethyl)-3-methyl-pentanamide,
1-benzylpiperidin-4-amine, (2R)-2-amino-3-ethylsulfanyl-propanoic
acid,
2-amino-N-[2-(2,5-dioxopiperazin-1-yl)-2-oxo-ethyl]propanamide,
2-amino-3-(1H-imidazol-4-yl)propanamide,
2-amino-N-(2-naphthyl)acetamide,
(2-amino-6-methyl-phenyl)-phenyl-methanone,
3-[2-(2-aminoethylamino)ethylamino]propanenitrile,
2-amino-1-(3-bromophenyl)ethanone,
(1,1-dioxothiolan-3-yl)methanamine, 2,4,6-tritert-butylaniline,
N1,N4-bis(4-amino-2-chloro-phenyl)terephthalamide,
4-[(3,4-diaminophenyl)methyl]benzene-1,2-diamine,
5-methoxy-2-methyl-1,3-benzothiazol-6-amine,
2-(2-methyl-5-nitro-imidazol-1-yl)ethanamine,
1-bromonaphthalen-2-amine, 4-amino-2,6-dibromo-benzenesulfonamide,
N'-[(E)-(2-aminophenyl)methyleneamino]-N-(4-chloro-3-nitro-phenyl)oxamide-
, 2-bromo-4,5-dimethyl-aniline, ethyl
2-[(4-amino-3-nitro-benzoyl)amino]-4,5,6,7-tetrahydrobenzothiophene-3-car-
boxylate, 4-amino-2-morpholinosulfonyl-phenol,
4-[(4-amino-3,5-diethyl-phenyl)methyl]-2,6-diethyl-aniline,
5-[1-(3-amino-4-methyl-phenyl)-2,2,2-trifluoro-1-(trifluoromethyl)ethyl]--
2-methyl-aniline, 4-pyridylmethanamine,
2-phenylbenzotriazole-4,5-diamine,
5-amino-2-hydroxy-N,N-dimethyl-benzenesulfonamide, methyl
2-amino-3-phenyl-propanoate,
4-amino-N-[4-[6-[(4-aminobenzoyl)amino]-7-chloro-1H-benzimidazol-2-yl]phe-
nyl]benzamide, 3-chloro-4-(2-naphthyloxy)aniline,
2-bromo-6-(difluoromethylsulfonyl)-4-nitro-aniline,
5-(4-aminophenoxy)-2-(1-naphthyl)isoindoline-1,3-dione,
5-(3-aminophenoxy)-2-(1-naphthyl)isoindoline-1,3-dione,
7-[3-(aminomethyl)-1-piperidyl]-1-cyclopropyl-8-methoxy-4-oxo-quinoline-3-
-carboxylic acid,
7-[3-(1-amino-1-methyl-ethyl)-1-piperidyl]-1-cyclopropyl-8-methoxy-4-oxo--
quinoline-3-carboxylic acid,
N-(3-amino-4-chloro-phenyl)-4,4-dimethyl-3-oxo-pentanamide,
(4-aminophenyl)-(4-fluorophenyl)methanone,
2-(5-fluoro-1H-indol-3-yl)ethanamine,
N1-(4-methoxyphenyl)benzene-1,4-diamine,
2-nitro-5-piperazin-1-yl-aniline,
5-(4-methylpiperazin-1-yl)-2-nitro-aniline,
2-amino-N--[(Z)-1-(4-chlorophenyl)ethylideneamino]benzamide,
3-amino-N-(2-amino-5-methyl-phenyl)-N-benzyl-benzamide,
1-[(Z)-1-(4-aminophenyl)ethylideneamino]-3-(m-tolyl)thiourea,
2-amino-4-cyclopropyl-6-(4-methoxyphenyl)benzene-1,3-dicarbonitrile,
2-(2-naphthyl)-1,3-benzoxazol-5-amine,
N-[(E)-1-(4-aminophenyl)ethylideneamino]furan-2-carboxamide,
4-(4-aminophenyl)thiazol-2-amine,
(2R)-2-acetamido-6-[[(2R)-2-aminobutanoyl]amino]-N-[[3-(trifluoromethyl)p-
henyl]methyl]hexanamide,
(4S)-5-[[(5R)-5-acetamido-6-oxo-6-(propylamino)hexyl]amino]-4-amino-5-oxo-
-pentanoic acid,
N-[(1R)-5-[[4-(aminomethyl)cyclohexanecarbonyl]amino]-1-[[(2R)-2-hydroxyp-
ropyl]carbamoyl]pentyl]thiophene-2-carboxamide,
N-[(1R)-1-(allylcarbamoyl)-5-[(4-aminobenzoyl)amino]pentyl]thiophene-2-ca-
rboxamide,
(4S)-4-amino-5-oxo-5-[[(5R)-6-oxo-6-[2-(2-thienyl)ethylamino]-5-
-(thiophene-2-carbonylamino)hexyl]amino]pentanoic acid,
2-[(6-amino-1,3-benzothiazol-2-yl)sulfanyl]-N-(2-fluorophenyl)acetamide,
N-(5-amino-2-methoxy-phenyl)-2,4-dichloro-benzamide,
N-(6-amino-4-methyl-1,3-benzothiazol-2-yl)acetamide,
3-amino-N'-[2-(2-naphthyloxy)acetyl]-5-nitro-benzohydrazide,
2-(2-aminophenyl)sulfanyl-N-[3,5-bis(trifluoromethyl)phenyl]-2-phenyl-ace-
tamide, ethyl
2-[[2-[2-[[2-amino-3-(4-hydroxyphenyl)propanoyl]amino]propanoylamino]acet-
yl]amino]acetate, 2-amino-5-chloro-N-(4-pyridylmethyl)benzamide,
8-nitronaphthalen-1-amine, 2-amino-3-cyclopropyl-propanoic acid,
2-(2-isopropyl-5-methyl-phenoxy)ethanamine,
2-amino-N-[(E)-1-(2-hydroxyphenyl)ethylideneamino]benzamide,
(2R)-2-amino-3-benzhydrylsulfanyl-propanoic acid, tert-butyl
2-aminopropanoate,
2-[4-(1-ethylpropyl)phenoxy]-5-(trifluoromethyl)aniline,
N1-methylbenzene-1,3-diamine,
1-(4-aminophenyl)sulfanyl-3-(diethylamino)propan-2-ol,
N-(4-aminophenyl)-2,2-dimethyl-propanamide,
2-amino-3-(4-nitrophenyl)butanoic acid,
2-(2-amino-5-bromo-phenyl)-4-methyl-benzo[g]quinoxalin-3-one,
N-[3-[(2-aminophenyl)methylamino]-1-methyl-3-oxo-propyl]-2-phenyl-quinoli-
ne-4-carboxamide,
N-[2-[(2-aminophenyl)methylamino]-2-oxo-1-phenyl-ethyl]-2-phenyl-quinolin-
e-4-carboxamide,
(5S)-5-(4-aminobutyl)-3-[4-(o-tolyl)phenyl]imidazolidine-2,4-dione,
(5S)-5-(4-aminobutyl)-3-[4-(benzothiophen-2-yl)-1-naphthyl]-2-thioxo-imid-
azolidin-4-one, 2-amino-4,6-ditert-butyl-phenol,
5-(aminomethyl)-2,4-dimethyl-pyridin-3-amine,
3-amino-N-[5-hydroxy-1-(2,4,6-trichlorophenyl)pyrazol-3-yl]benzamide,
(2R)-2-amino-3-(4-fluorophenyl)-N-[4-guanidino-1-(1-piperidylmethyl)butyl-
]propanamide,
3-[[2-[2-(3-aminopropylcarbamoyl)phenyl]benzoyl]-[(2,5-difluorophenyl)met-
hyl]amino]propanoic acid,
N-[(4-acetamidophenyl)methyl]-N-(3-amino-2,2-dimethyl-propyl)-2-(4-ethylp-
henyl)pyridine-4-carboxamide,
N-(3-aminopropyl)-2-(4-ethylphenyl)-N-[(3,4,5-trimethoxyphenyl)methyl]pyr-
idine-4-carboxamide,
N-(2-aminoethyl)-5-(4-fluorophenyl)-N-(2-pyridylmethyl)pyridine-3-carboxa-
mide,
N-[[4-(aminomethyl)phenyl]methyl]-5-(1-naphthyl)-N-(2-pyridylmethyl)-
pyridine-3-carboxamide,
2-(3-acetylphenyl)-N-(3-aminopropyl)-N-(2,3-dihydro-1,4-benzodioxin-6-ylm-
ethyl)pyridine-4-carboxamide,
2-[(4S,5R)-2-[(1R)-1-amino-2-(4-fluorophenyl)ethyl]-5-(2-naphthyl)tetrahy-
dropyran-4-yl]acetonitrile,
(2R)-2-amino-1-[(2S,4R)-4-benzyloxy-2-[2-(1,2,4-triazol-4-yl)ethyl]pyrrol-
idin-1-yl]-3-(4-fluorophenyl)propan-1-one,
(2R)-2-amino-3-phenyl-1-[4-phenyl-4-(1,2,4-triazol-1-ylmethyl)-1-piperidy-
l]propan-1-one, N'-cyclododecylethane-1,2-diamine,
7-[2-[(2-amino-2-methyl-propyl)amino]pyrimidin-4-yl]-6-(4-fluorophenyl)-2-
,3-dihydro-1H-pyrazolo[1,2-a]pyrazol-5-one,
2,3,4,5-tetrahydro-1-benzothiepin-5-amine,
5-[(2R,3R,4S)-3-amino-4-(methoxycarbonylamino)tetrahydrothiophen-2-yl]pen-
tanoic acid,
3-(2-aminophenyl)sulfanyl-3-(3,4-dichlorophenyl)-1-phenyl-propan-1-one,
and pharmaceutically acceptable salts thereof.
[0175] The primary amine compounds used in methods described herein
can be administered to the subject to treat the ocular disorder
(e.g., macular degeneration or Stargardt disease, geographic
atrophy) using standard delivery methods including, for example,
ophthalmic, topical, parenteral, subcutaneous, intravenous,
intraarticular, intrathecal, intramuscular, intraperitoneal,
intradermal injections, or by transdermal, buccal, oromucosal, oral
routes or via inhalation. The particular approach and dosage used
for a particular subject depends on several factors including, for
example, the general health, weight, and age of the subject. Based
on factors such as these, a medical practitioner can select an
appropriate approach to treatment.
[0176] Treatment according to the method described herein can be
altered, stopped, or re-initiated in a subject depending on the
status of ocular disorder. Treatment can be carried out as
intervals determined to be appropriate by those skilled in the art.
For example, the administration can be carried out 1, 2, 3, or 4
times a day. In another embodiment, the primary amine compound can
be administered after induction of macular degeneration has
occurred.
[0177] The treatment methods can include administering to the
subject a therapeutically effective amount of the primary amine
compound. Determination of a therapeutically effective amount is
within the capability of those skilled in the art. The exact
formulation, route of administration, and dosage can be chosen by
the individual physician in view of the subject's condition.
[0178] Formulation of pharmaceutical compounds for use in the modes
of administration noted above (and others) are described, for
example, in Remington's Pharmaceutical Sciences (18th edition), ed.
A. Gennaro, 1990, Mack Publishing Company, Easton, Pa. (also see,
e.g., M. J. Rathbone, ed., Oral Mucosal Drug Delivery, Drugs and
the Pharmaceutical Sciences Series, Marcel Dekker, Inc., N.Y.,
U.S.A., 1996; M. J. Rathbone et al., eds., Modified-Release Drug
Delivery Technology, Drugs and the Pharmaceutical Sciences Series,
Marcel Dekker, Inc., N.Y., U.S.A., 2003; Ghosh et al., eds., Drug
Delivery to the Oral Cavity, Drugs and the Pharmaceutical Sciences
Series, Marcel Dekker, Inc., N.Y. U.S.A., 1999.
[0179] In one example, the primary amine compound can be provided
in an ophthalmic preparation that can be administered to the
subject's eye. The ophthalmic preparation can contain the primary
amine compound in a pharmaceutically acceptable solution,
suspension or ointment. Some variations in concentration will
necessarily occur, depending on the particular primary amine
compound employed, the condition of the subject to be treated and
the like, and the person responsible for treatment will determine
the most suitable concentration for the individual subject. The
ophthalmic preparation can be in the form of a sterile aqueous
solution containing, if desired, additional ingredients, for
example, preservatives, buffers, tonicity agents, antioxidants,
stabilizers, nonionic wetting or clarifying agents, and viscosity
increasing agents.
[0180] Examples of preservatives for use in such a solution include
benzalkonium chloride, benzethonium chloride, chlorobutanol,
thimerosal and the like. Examples of buffers include boric acid,
sodium and potassium bicarbonate, sodium and potassium borates,
sodium and potassium carbonate, sodium acetate, and sodium
biphosphate, in amounts sufficient to maintain the pH at between
about pH 6 and about pH 8, and for example, between about pH 7 and
about pH 7.5. Examples of tonicity agents are dextran 40, dextran
70, dextrose, glycerin, potassium chloride, propylene glycol, and
sodium chloride.
[0181] Examples of antioxidants and stabilizers include sodium
bisulfite, sodium metabisulfite, sodium thiosulfite, and thiourea.
Examples of wetting and clarifying agents include polysorbate 80,
polysorbate 20, poloxamer 282 and tyloxapol. Examples of
viscosity-increasing agents include gelatin, glycerin,
hydroxyethylcellulose, hydroxmethylpropylcellulose, lanolin,
methylcellulose, petrolatum, polyethylene glycol, polyvinyl
alcohol, polyvinylpyrrolidone, and carboxymethylcellulose. The
ophthalmic preparation will be administered topically to the eye of
the subject in need of treatment by conventional methods, for
example, in the form of drops or by bathing the eye in the
ophthalmic solution.
[0182] The primary amine compound can also be formulated for
topical administration through the skin. "Topical delivery systems"
also include transdermal patches containing the ingredient to be
administered. Delivery through the skin can further be achieved by
iontophoresis or electrotransport, if desired.
[0183] Formulations for topical administration to the skin can
include, for example, ointments, creams, gels and pastes comprising
the primary amine compound in a pharmaceutical acceptable carrier.
The formulation of the primary amine compound for topical use
includes the preparation of oleaginous or water-soluble ointment
bases, as is well known to those in the art. For example, these
formulations may include vegetable oils, animal fats, and, for
example, semisolid hydrocarbons obtained from petroleum. Particular
components used may include white ointment, yellow ointment, cetyl
esters wax, oleic acid, olive oil, paraffin, petrolatum, white
petrolatum, spermaceti, starch glycerite, white wax, yellow wax,
lanolin, anhydrous lanolin and glyceryl monostearate. Various
water-soluble ointment bases may also be used, including glycol
ethers and derivatives, polyethylene glycols, polyoxyl 40 stearate
and polysorbates.
[0184] Subjects affected with or at risk of macular degeneration,
which are not readily accessible or suitable for ophthalmic (e.g.
eye-drops) and/or topical administration, can be treated by a
systemic approach, such as intravenous infusion. For example, the
primary amine compound can be administered at a low dosage by
continuous intravenous infusion. In another example, in which a
patient requires longer-term care, the primary amine compound can
be administered intermittently (e.g., every 12-24 hours). In a
variation of this approach, the initial or loading dose can be
followed by maintenance doses that are less than, (e.g., half) the
loading dose or by continuous infusion. The duration of such
treatment can be determined by those having skill in the art, based
on factors, for example, the severity of the condition and the
observation of improvements.
[0185] When administering the primary amine compound to the subject
by intravenous infusion, devices and equipment (e.g., catheters,
such as central or peripheral venous catheters, tubing, drip
chambers, flashback bulbs, injection Y sites, stopcocks, and
infusion bags) can be used that are compatible with the primary
amine compound.
[0186] As discussed above, the primary amine compounds may be
administered to a subject in order to treat or prevent macular
degeneration and other forms of retinal disease whose etiology
involves aberrant all-trans-RAL clearance. Other diseases,
disorders, or conditions characterized by aberrant all-trans-RAL
may be similarly treated.
[0187] In one embodiment, a subject is diagnosed as having symptoms
of macular degeneration, and then a disclosed compound is
administered. In another embodiment, a subject may be identified as
being at risk for developing macular degeneration (risk factors
include a history of smoking, age, female gender, and family
history), and then a disclosed compound is administered. In another
embodiment, a subject may have dry AMD in both eye, and then a
disclosed compound is administered. In another embodiment, a
subject may have wet AMD in one eye but dry AMD in the other eye,
and then a disclosed compound is administered. In yet another
embodiment, a subject may be diagnosed as having Stargardt disease
and then a disclosed compound is administered. In another
embodiment, a subject is diagnosed as having symptoms of other
forms of retinal disease whose etiology involves aberrant
all-trans-RAL clearance, such as geographic atrophy (GA), and then
the compound is administered. In another embodiment, a subject may
be identified as being at risk for developing other forms of
retinal disease whose etiology involves all-trans-RAL clearance,
and then the disclosed compound is administered. In some
embodiments, a compound is administered prophylactically. In some
embodiments, a subject has been diagnosed as having the disease
before retinal damage is apparent. In some embodiments, a human
subject may know that he or she is in need of the macular
generation treatment or prevention.
[0188] In some embodiments, a subject may be monitored for the
extent of macular degeneration. A subject may be monitored in a
variety of ways, such as by eye examination, dilated eye
examination, fundoscopic examination, visual acuity test, and/or
biopsy. Monitoring can be performed at a variety of times. For
example, a subject may be monitored after a compound is
administered. The monitoring can occur, for example, one day, one
week, two weeks, one month, two months, six months, one year, two
years, five years, or any other time period after the first
administration of a compound. A subject can be repeatedly
monitored. In some embodiments, the dose of a compound may be
altered in response to monitoring.
[0189] In some embodiments, the disclosed methods may be combined
with other methods for treating or preventing macular degeneration
or other forms of retinal disease whose etiology involves aberrant
all-trans-RAL clearance, such as photodynamic therapy. For example,
a patient may be treated with more than one therapy for one or more
diseases or disorders. For example, a patient may have one eye
afflicted with dry form AMD, which is treated with a compound of
the invention, and the other eye afflicted with wet form AMD, which
is treated with, e.g., photodynamic therapy.
[0190] In yet another embodiment, the primary amine compound
described herein can be administered as part of a combinatorial
therapy with additional therapeutic agents. The phrase
"combinatorial therapy" or "combination therapy" embraces the
administration of a primary amine compound, and one or more
therapeutic agents as part of a specific treatment regimen intended
to provide beneficial effect from the co-action of these
therapeutic agents. Administration of these therapeutic agents in
combination typically is carried out over a defined period (usually
minutes, hours, days or weeks depending upon the combination
selected). "Combinatorial therapy" or "combination therapy" is
intended to embrace administration of these therapeutic agents in a
sequential manner, that is, wherein each therapeutic agent is
administered at a different time, as well as administration of
these therapeutic agents, or at least two of the therapeutic
agents, in a substantially simultaneous manner. Substantially
simultaneous administration can be accomplished, for example by
administering to the subject an individual dose having a fixed
ratio of each therapeutic agent or in multiple, individual doses
for each of the therapeutic agents. Sequential or substantially
simultaneous administration of each therapeutic agent can be
effected by any appropriate route including, but not limited to,
oral routes, intravenous routes, intramuscular routes, and direct
absorption through mucous membrane tissue. The therapeutic agents
can be administered by the same route or by different routes. The
sequence in which the therapeutic agents are administered is not
narrowly critical.
[0191] The invention is further illustrated by the following
example, which is not intended to limit the scope of the
claims.
Example 1
Formation of Schiff Base Between Retinal and Selected Amines
[0192] We tested 5 FDA approved drugs containing primary amino
groups to determine if they react with the aldehyde group of
all-trans-RAL to form conjugates (Schiff-base) under the
physiological conditions of the eye. The five FDA approved drugs
included Potaba, Paser, Seromycin, Cuprimine, and Lyrica.
[0193] Formation of conjugates (Schiff-base) between retinal and
primary amine compounds containing amino group (putative drugs) is
the key mechanism to control all-trans-RAL levels in the retina and
prevent retina degeneration. Stability of these conjugates is an
important factor that can determine biological activity of the
drugs. To standardize conditions of Schiff-base formation all
reactions were performed in 90% ethanol in water buffered with 0.1
M phosphate buffer, pH 7.0. Stock solution of retinal was made up
freshly in ethanol. Its final concentration was determined
spectrophotometricaly at 380 nm (.epsilon.=42,880). All procedures
were carried out in the dark. Schiff base formation was initiated
by addition of retinal stock solution to a 2 molar excess of tested
amine dissolved in the reaction buffer. The reaction mixture was
incubated for 1 h in room temperature.
[0194] Steady-state rate of Schiff-base formation was obtained from
absorbance changes at 380 and 440 nm (510 nm for aromatic amines)
corresponding to free retinal and protonated Schiff base,
respectively. To investigate stability of the given retinal
conjugates a mixture of HPLC purified Schiff base of selected
compound and phosphatidylocholine was dried down in SpeedVac. Then,
Schiff-base and lipids were overlaid with 0.1 M phosphate buffer
and sonicated immediately to form liposomes. The samples were
extracted with hexane following 30 min incubated in room
temperature. The breakdown of Schiff-base was monitored by HPLC
detection of free retinal extracted from the reaction mixture.
[0195] FIGS. 3-5 illustrate UV/Vis spectra for the FDA approved
drugs. FDA approved drugs that were active readily formed a Schiff
base with all-trans-RAL as evidenced by a substantial differential
between spectra of unprotonated and protonated Schiff base.
Whereas, compounds that were inactive did not readily form a Schiff
Base with all-trans-RAL and showed little differential in the
spectra. Of the FDA approved drugs tested, Paser, Seromycin, and
Lyrica readily formed stable Schiff-bases with all-trans-RAL. In
contrast, Potaba and Curprimine did not readily form stable
Schiff-bases with all-trans-RAL.
Example 2
[0196] We tested 21 drugs containing primary amino groups to
determine if they react with the aldehyde group of all-trans-RAL to
reduce its toxic levels in the eye after exposure to light and
prevent its condensation into toxic conjugates. The drugs included
Flumadine, Nameda, Potaba, Dapsone, Paser, Luvox, Seromycin,
Aminohippurate Sodium, Cuprimine, Januvia, Primaxin I.M., Prinivil,
Sulfamylon, Exforge, Stalevo, Sodium Diuril, Lyrica
((S)-3-(aminomethyl)-5-methylhexanoic acid),
(R)-3-(aminomethyl)-5-methylhexanoic acid), Asacol, Tamiflu, and
Rilutek. For these experiments, we used ABCA4.sup.-/-/RDH8.sup.-/-
mice as models for Stargardt's disease and AMD. We employed
analytical methods for determining the pharmacodynamics and
pharmacokinetics of candidate drugs including: HPLC/MS/MS for
analysis of retinoids and histological sections and OCT for
assessment of retinal pathology. The relative amount of retinoids
and their composition strongly correlates with the health status of
the retina. FDA approved drugs were determined effective in
treating retinal degeneration if upon administration to
ABCA4.sup.-/-/RDH8.sup.-/- mice, the mice showed optical coherence
tomography score of at least about 2.5 and the drug increased
11-cis-retinal amount at least about 30% in comparison to untreated
control animal. Methodologies for performing the analysis on
ABCA4.sup.-/-/RDH8.sup.-/- mice is described below.
Animals
[0197] Rdh8.sup.-/- mice were generated and genotyped as previously
described in J. Biol. Chem. 280, 188-18832 (2005). Abca4.sup.-/-
mice also were generated by standard procedures (Ingenious
Targeting, Inc., Stony Brook, N.Y.). The targeting vector was
constructed by replacing exon 1 with the neo cassette as described
by Cell, 98 13-23 (1999). No Immunoreactivity against ABCA4 was
detected in eye extracts from these mice by Immunocytochemistry or
Immunoblottmg. Abca4.sup.-/- mice were maintained with either
pigmented 129Sv/Ev or C57BL/6 mixed backgrounds, and their siblings
were used for most experiments. Rdh8.sup.-/-Abca4.sup.-/- mice were
established by crossbreeding Abca4.sup.-/- mice with Rdh8.sup.-/-
mice. Genotyping or mice was carried out by PCR with primers ABCR1
(5'-gcccagtggtcgatctgtctagc-3') (SEQ ID NO: 1) d ABCR2
(5'-cacaaaggccgctaggaccacg-3') (SEQ ID NO: 2) for wild type (WT)
(619 bp) and A0 (5'-ccacagcacacatcagcatttctcc-3') (SEQ ID NO: 3)
and N1 (5'-tgcgaggccagaggccacttgtgtagc-3') (SEQ ID NO: 4) for
targeted deletion (455 bp). PCR products were cloned and sequenced
to verify their identities. Rdh8.sup.-/-Abca4.sup.-/- mice were
fertile and showed no obvious developmental abnormalities.
Extraction and Hplc Analysis of Non-Polar Retinoids
[0198] All experimental procedures related to extraction,
derivatization, and separation of retinoids from dissected mouse
eyes were carried out under dim red light. Two whole mouse eyes
were homogenized in 1 ml of 50 mM phosphate buffer, pH 7.0,
containing 50% ethanol and 10 mM NH.sub.2OH. Ice-cold methanol (1
ml) was added to the homogenates 20 min after incubation at room
temperature (RT), non-polar retinoids were extracted twice with 4
ml of hexane. The organic phase was collected, dried down in a
SpeedVac and re-solubilized in 0.3 ml of hexane. Three main classes
of retinoids (retinyl esters, retinal oximes, and retinols) as well
as their geometrical isomers was separated in single run by normal
phase HPLC by using an Agilent Si, 5 .mu.m, 4.5.times.250 mm column
and a stepwise gradient of ethylacetate in hexane (0.5% for 15 min,
and 6% for up to 60 min) at a flow rate of 1.4 ml/min (FIG. 6).
Retinoids were detected at 325 nm (retinyl esters and retinols) and
350 nm (retinyl oximes) with a diode array detector. Those of
interest were quantified based on the areas under their peaks
calculated with the help of HP Chemstation A.03.03 software and
compared with areas calculated based on known amounts of synthetic
standards plotted as a standard curve.
Mass Spectrometry of Retinoids
[0199] A complementary technique, mass spectrometry, was used to
detect, identify, and quantify retinoids and their derivatives in
eye tissue. The conjugated polyene chain of retinoids contributes
to relatively strong light absorption at UV and visible
wavelengths. Thus, absorbance spectra provided information about
the number of conjugated double bonds. Moreover, slight differences
in wavelengths of maximum absorbance and shapes of the spectra
permitted precise identification of retinoid isomers. However, a
limitation of this method is the low selectivity of its UV-Vis
absorbance, which mandates carefully designed chromatographic
conditions and precise identification of the compounds being
analyzed. This analysis can become especially challenging when
multiple geometric isomers of retinoids at low abundance (less than
3 pmols/eye) or unidentified compounds are present. Thus, we used
an alternative technique, mass spectrometry combined with high
performance liquid chromatography (LC-MS) to address this issue.
Mass spectra of retinoids was acquired by using a LXQ high
throughput linear ion trap mass spectrometer (Thermo Scientific,
Waltham, Mass.) connected with an Agilent 1100 HPLC system and
interfaced with an atmospheric pressure chemical ionization (APCI)
source. The APCI source in a positive ionization mode is chosen for
LC-MS methodology because of its wide dynamic range and capacity to
operate at the high flow rates required for HPLC retinoid
separation. The greatest advantage of LC-MS is its sensitivity that
reaches the limits of retinoid detection and quantification in the
10 to 50 fmol and 20 to 200 fmol ranges, respectively. Moreover,
this LXQ instrument has capability to perform MSn analyses that
provide definitive structural identification.
Detection and Quantification of Retinal Amine Condensation Products
in Mouse Eye
[0200] Eyes of mice treated with compounds containing primary
amines were homogenized in Tris/HCl buffer, pH 9.0, in 50% methanol
and extracted with hexane or ethyl acetate depending of the
polarity, dried down and resuspended in acetonitrile. After
centrifugation, extracted compounds in the supernatant were
separated by reverse phase HPLC chromatography (Agilent Zorbax
Eclipse XBD C18, 5 .mu.m, 4.6.times.150 mm column) with a linear
gradient of water in acetonitrile (50-100%) for 20 min at a fixed
flow rate of 1.5 ml/min. Retinal conjugates were detected and
identified with a LXQ mass spectrometer equipped with an APCI
source. MS scans were recorded in a SIM mode for each individual
compound (FIG. 7 M3). The identity of detected adducts will be
confirmed based on their MS.sup.2 spectra. Amounts of retinal-amine
conjugates will be quantified with the aid of isotopically labeled
synthetic standards added prior to extraction.
Ultra-High Resolution Spectral-Domain Optical Coherence Tomography
(SD-OCT) Imaging
[0201] Although analytical and histological methods provide
exhaustive characterization of retina, they cannot be performed in
vivo. To reduce the number of sacrificed animals and time required
for analysis, SD-OCT from Bioptigen (Research Triangle Park, N.C.)
was employed for in vivo imaging of mouse retinas. Bioptigen OCT
systems utilize a narrow single-mode beam from a wide bandwidth
light source to probe the structure of retina at a higher
resolution (2.0 .mu.m) than normal OCT systems. Mice were
anesthetized by intraperitoneal (IP) injection of 20 .mu.l/g bw of
6 mg/ml ketamine and 0.44 mg/ml xylazine diluted with 10 mM sodium
phosphate, pH 7.2, containing 100 mM NaCl. Pupils were dilated with
1% tropicamide. In vivo SD-OCT images were obtained from both eyes.
Four pictures acquired in a B-scan mode were used to construct the
final averaged images (FIG. 8). SD-OCT imaging enabled us to
identify early pathological changes in the retina and monitor
progression or amelioration/prevention of pathological lesions
quantitatively under various therapeutic regimens in the same live
animal at a resolution comparable to that obtained by current
histopathological methods that employ cross sections of the
retina.
Grading with OCT
[0202] In vivo retinal structures of Rdh8.sup.-/-Abca4.sup.-/- mice
are imaged by SD-OCT 7 days after 10,000 lux illumination for 30
min at 4 weeks of age. FIG. 9 illustrates Rdh8.sup.-/-Abca4.sup.-/-
mouse without light shows healthy retina, whereas light with 10,000
lux for 30 min cause severe retinal degeneration (left panel).
Preventive effects of compounds in retinal morphology are indicated
as OCT score (right panel). [0203] Score 5: no retinal degeneration
[0204] Score 4: regional retinal degeneration (less than 1000 .mu.m
width) [0205] Score 3: widely observed retinal degeneration (more
than 1000 .mu.m width) with reflection of ELM [0206] Score 2:
widely observed retinal degeneration (more than 1000 .mu.m width)
without reflection of ELM [0207] Score 1: severe retinal
degeneration without reflection from photoreceptors as well as
light exposed Rdh8.sup.-/-Abca4.sup.-/- mice (see left panel)
Results
[0208] The following Table lists the OCT score and 11 cis-retinal
area of ABCA4.sup.-/-/RDH8.sup.-/- treated with Flumadine, Nameda,
Potaba, Dapsone, Paser, Luvox, Seromycin, Aminohippurate Sodium,
Cuprimine, Januvia, Primaxin I.M., Prinivil, Sulfamylon, Exforge,
Stalevo, Sodium Diuril, Lyrica,
(R)-3-(aminomethyl)-5-methylhexanoic acid, Asacol, Tamiflu, and
Rilutek. As noted in the Table, ABCA4.sup.-/-/RDH8.sup.-/- mice
treated with either Flumadine, Dapsone, Paser, Luvox, Seromycin,
Januvia, Sulfamylon, Exforge, Sodium Diuril, Lyrica,
(R)-3-(aminomethyl)-5-methylhexanoic acid, Asacol, or Rilutek had a
optical coherence tomography score of at least about 2.5 and the
drug increased 11-cis-retinal amount at least about 30% in
comparison to untreated control animal and were effective in
treating retinal degeneration in the mice. It is noted that the
three FDA approved drugs that readily formed Schiff bases in
Example 1 with all-trans-RAL under simulated physiological
conditions had a optical coherence tomography score of at least
about 2.5, increased 11-cis-retinal amount at least about 30% in
comparison to untreated control animal, and were effective in
treating retinal degeneration in the mice. In contrast, the two FDA
approved drugs that did not readily formed Schiff bases in Example
1 with all-trans-RAL under simulated physiological conditions had
an optical coherence tomography score below 2.5, did not increase
11-cis-retinal amount at least about 30% in comparison to untreated
control animal, and were ineffective in treating retinal
degeneration in the mice.
TABLE-US-00001 TABLE Score 11cRAL OCT (area) 1 Flumadine
.gtoreq.2.5 135.3 2 Nameda 1.7 97.6 3 Potaba 2.3 90.6 4 Dapsone
.gtoreq.2.5 156.5 5 Paser .gtoreq.2.5 151.9 6 Luvox .gtoreq.2.5
175.5 7 Seromycin .gtoreq.2.5 159.9 8 Aminohippurate 1 109.4 Sodium
9 Cuprimine 1.7 118.5 10 Januvia .gtoreq.2.5 126.8 11 Primaxin I.M.
1.7 108.1 12 Prinivil 2.0 132.7 13 Sulfamylon .gtoreq.2.5 166.1 14
Exforge .gtoreq.2.5 173.8 15 Stalevo 1.3 86.7 16 Sodium Diuril
.gtoreq.2.5 139.7 17 Lyrica .gtoreq.2.5 166.5 (S)-3-(aminomethyl)-
5-methylhexanoic acid 18 (R)-3-(aminomethyl)- .gtoreq.2.5 164.8
5-methylhexanoic acid 19 Asacol .gtoreq.2.5 140.2 20 Tamiflu 1.3
87.3 21 Rilutek .gtoreq.2.5 158.2 22 No Treatment 1 88.2
[0209] From the above description of the invention, those skilled
in the art will perceive improvements, changes and modifications.
Such improvements, changes and modifications within the skill of
the art are intended to be covered by the appended claims. All
references, publications, and patents cited in the present
application are herein incorporated by reference in their
entirety.
* * * * *