U.S. patent application number 12/063177 was filed with the patent office on 2008-10-30 for management of ophthalmologic disorders, including macular degeneration.
Invention is credited to Robert R. Rando.
Application Number | 20080269324 12/063177 |
Document ID | / |
Family ID | 37711019 |
Filed Date | 2008-10-30 |
United States Patent
Application |
20080269324 |
Kind Code |
A1 |
Rando; Robert R. |
October 30, 2008 |
Management of Ophthalmologic Disorders, Including Macular
Degeneration
Abstract
A drag may be used in the preparation of a medicament for the
treatment or prevention of an ophthalmologic disorder, wherein the
drug inhibits, antagonizes, or short-circuits the visual cycle at a
step of the visual cycle that occurs outside a disc of a rod
photoreceptor cell.
Inventors: |
Rando; Robert R.;
(Brookline, MA) |
Correspondence
Address: |
FOLEY HOAG, LLP;PATENT GROUP (w/HUV HMV)
155 SEAPORT BLVD.
BOSTON
MA
02210-2600
US
|
Family ID: |
37711019 |
Appl. No.: |
12/063177 |
Filed: |
August 7, 2006 |
PCT Filed: |
August 7, 2006 |
PCT NO: |
PCT/US2006/030892 |
371 Date: |
June 30, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11199594 |
Aug 8, 2005 |
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12063177 |
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Current U.S.
Class: |
514/478 ;
514/512; 514/546; 514/627; 560/129; 564/204; 564/207; 568/417 |
Current CPC
Class: |
C07C 211/45 20130101;
C07C 403/12 20130101; C07C 403/16 20130101; C07C 49/203 20130101;
C07C 49/203 20130101; C07C 229/60 20130101; C07C 43/15 20130101;
C07C 233/09 20130101; C07B 2200/09 20130101; C07C 233/05 20130101;
C07D 203/08 20130101; C07C 233/25 20130101; C07C 69/24 20130101;
C07C 2601/14 20170501; C07C 243/38 20130101; C07C 403/10 20130101;
A61P 27/02 20180101; C07C 45/68 20130101; C07C 403/18 20130101;
C07C 229/56 20130101; C07C 217/86 20130101; C07C 251/40 20130101;
C07C 403/08 20130101; C07D 303/08 20130101; C07C 2601/16 20170501;
C07C 43/162 20130101; C07C 57/26 20130101; C07C 403/20 20130101;
C07C 45/68 20130101 |
Class at
Publication: |
514/478 ;
564/204; 564/207; 560/129; 568/417; 514/546; 514/512; 514/627 |
International
Class: |
A61K 31/16 20060101
A61K031/16; C07C 233/02 20060101 C07C233/02; C07C 69/587 20060101
C07C069/587; C07C 49/203 20060101 C07C049/203; C07C 233/09 20060101
C07C233/09; A61P 27/02 20060101 A61P027/02; A61K 31/22 20060101
A61K031/22; A61K 31/27 20060101 A61K031/27; A61K 31/265 20060101
A61K031/265 |
Goverment Interests
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] Support for research leading to subject matter disclosed in
this application was provided in part by the National Institutes of
Health Grant Nos. RO1-EY-04096 and/or RO1-EY-015425. Accordingly,
the United States Government has certain rights with respect to
subject matter of this application.
Claims
1. A compound of formula XI: ##STR00149## wherein, independently
for each occurrence, n is 0 to 10 inclusive; R.sup.1 is hydrogen or
alkyl; R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; Z is cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, aralkyl, --C(.dbd.O)R.sub.b, or
--(CH.sub.2).sub.pR.sub.b; p is 0 to 20 inclusive; R.sub.a is
hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,
or aralkyl; R.sub.b is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; and denotes a single bond
or a trans double bond; provided that the compound is not
N-(4-hydroxyphenyl)retinamide.
2. The compound of claim 1, wherein R.sup.1 is hydrogen or
methyl.
3. The compound of claim 1, wherein Z is aryl.
4. The compound of claim 1, wherein R.sub.a is hydrogen.
5. A compound of claim 1, wherein said compound of formula XI is a
compound of formula XIa: ##STR00150## wherein, independently for
each occurrence, n is 0 to 10 inclusive; R.sup.1 is hydrogen or
alkyl; R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; R.sup.3, R.sup.4, R.sup.5,
R.sup.6 and R.sup.7 are hydrogen, halogen, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, or sulfoxido; R.sub.a is hydrogen, alkyl, cycloalkyl,
alkenyl, cycloalkenyl, alkynyl, aryl, or aralkyl; and denotes a
single bond or a trans double bond; provided that the compound is
not N-(4-hydroxyphenyl)retinamide.
6. The compound of claim 5, wherein R.sup.1 is hydrogen or
methyl.
7. The compound of claim 5, wherein R.sub.a is hydrogen.
8. The compound of claim 5, wherein R.sup.3, R.sup.4, R.sup.6 and
R.sup.7 are hydrogen.
9. The compound of claim 5, wherein R.sup.5 is hydroxyl.
10. A compound of claim 1, wherein said compound of formula XI is a
compound of formula XIb: ##STR00151## wherein, independently for
each occurrence, n is 0 to 5 inclusive; R.sup.1 is hydrogen or
methyl; R.sup.2 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
##STR00152## R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7 and
R.sup.8 are hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, or sulfoxido; any two geminal R.sup.8 and the carbon to
which they are bound may represent C(.dbd.O); and R.sub.a is
hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,
or aralkyl; provided that the compound is not
N-(4-hydroxyphenyl)retinamide.
11. The compound of claim 10, wherein R.sup.1 is hydrogen or
methyl.
12. The compound of claim 10, wherein R.sub.a is hydrogen.
13. The compound of claim 10, wherein R.sup.3, R.sup.4, R.sup.6 and
R.sup.7 are hydrogen.
14. The compound of claim 10, wherein R.sup.5 is hydroxyl.
15. The compound of claim 10, wherein n is 1 to 3 inclusive.
16. The compound of claim 10, wherein R.sup.2 is ##STR00153##
17. The compound of claim 10, wherein R.sup.2 is ##STR00154##
18. The compound of claim 10, wherein R.sup.2 is ##STR00155##
19. A compound of formula IV: ##STR00156## wherein, independently
for each occurrence, n is 1, 2, 3 or 4; X is --O--, --NR.sub.a--,
--C(R.sub.b).sub.2- or --C(.dbd.O)--; Z is --C(.dbd.O)R.sub.b,
--OR.sub.b, --N(R.sub.b).sub.2, alkyl or haloalkyl; R.sub.a is
hydrogen, alkyl, haloalkyl, aryl or aralkyl; and R.sub.b is
hydrogen, alkyl, haloalkyl, aryl or aralkyl.
20. A compound having a structure represented by ##STR00157##
21. A formulation comprising a compound defined by claim 1, and a
second compound, different from the first compound, wherein said
second compound is selected from compounds of formula I, II, III,
IV, V, VI, VII, VIII, IX, or X.
22-27. (canceled)
28. A formulation comprising a compound defined by claim 19, and a
second compound, different from the first compound, wherein said
second compound is selected from compounds of formula I, II, III,
IV, V, VI, VII, VIII, IX, or X.
29. A formulation comprising a compound defined by claim 20, and a
second compound, different from the first compound, wherein said
second compound is selected from compounds of formula I, II, III,
IV, V, VI, VII, VIII, IX, or X.
30. A method for treating or preventing an opthalmologic disorder
in a subject comprising administering to a subject a
pharmaceutically acceptable amount of a compound of claim 1.
31. A method for treating or preventing an opthalmologic disorder
in a subject comprising administering to a subject a
pharmaceutically acceptable amount of a compound of claim 19.
32. A method for treating or preventing an opthalmologic disorder
in a subject comprising administering to a subject a
pharmaceutically acceptable amount of a compound of claim 20.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims priority to U.S. patent application
Ser. No. 11/199,594, filed Aug. 8, 2005; which is a
continuation-in-part of International Application No.
PCT/US2005/004990, filed Feb. 17, 2005, which claims the benefit of
U.S. Provisional Patent Application Ser. No. 60/545,456, filed Feb.
17, 2004; U.S. Provisional Patent Application Ser. No. 60/567,604,
filed May 3, 2004; and U.S. Provisional Patent Application Ser. No.
60/578,324, filed Jun. 9, 2004. All aforementioned applications are
hereby incorporated herein by reference in their entirety.
INTRODUCTION
[0003] Age related diseases of vision are an ever-increasing health
problem in industrial societies. Age related macular degeneration
(AMD) affects millions of persons worldwide and is a leading cause
of vision loss and blindness in ageing populations. In this
disease, daytime vision (cone dominated vision) degrades with time
because cone photoreceptors, which are concentrated in the foveal
region of the retina, die. The incidence of this disease increases
from less than 10% of the population 50 years of age to over 30% at
75 and continues upwards past this age. The onset of the disease
has been correlated with the accumulation of complex and toxic
biochemicals in and around the retinal pigment epithelium (RPE) and
lipofuscin in the RPE. The accumulation of these retinotoxic
mixtures is one of the most important known risk factors in the
etiology of AMD. The RPE forms part of the retinal-blood barrier
and also supports the function of photoreceptor cells, including
rods and cones. Among other activities, the RPE routinely
phagocytoses spent outer segments of rod cells. In at least some
forms of macular degeneration, accumulation of lipofuscin in the
RPE is due in part to this phagocytosis. Retinotoxic compounds form
in the discs of rod photoreceptor outer segments. Consequently, the
retinotoxic compounds in the disc are brought into the RPE, where
they impair further phagocytosis of outer segments and cause
apoptosis of the RPE. Photoreceptors cells, including cone cells
essential for daytime vision, then die, denuded of RPE support.
[0004] One of the retinotoxic compounds formed in the discs of rod
outer segments is N-retinylidene-N-retinylethanolamine (A.sub.2E),
which is an important component of the retinotoxic lipofuscins.
A.sub.2E is normally formed in the discs but in such small amounts
that it does not impair RPE function upon phagocytosis. However, in
certain pathological conditions, so much A.sub.2E can accumulate in
the disc that the RPE is "poisoned" when the outer segment is
phagocytosed.
[0005] A.sub.2E is produced from all-trans-retinal, one of the
intermediates of the rod cell visual cycle. During the normal
visual cycle (summarized in FIG. 1), all-trans-retinal is produced
inside rod outer-segment discs. The all-trans-retinal can react
with phosphatidylethanolamine (PE), a component of the disc
membrane, to form N-retinylidene-PE. Rim protein (RmP), an
ATP-binding cassette transporter located in the membranes of rod
outer-segment discs, then transports all-trans-retinal and/or
N-retinylidene-PE out of the disc and into rod outer-segment
cytoplasm. The environment there favors hydrolysis of the
N-retinylidene-PE. The all-trans-retinal is reduced to
all-trans-retinol in the rod cytoplasm. The all-trans-retinol then
crosses the rod outer-segment plasma membrane into the
extracellular space and is taken up by cells of the retinal pigment
epithelium (RPE). The all-trans-retinol is converted through a
series of reactions to 11-cis-retinal, which returns to the
photoreceptor and continues in the visual cycle. However, defects
in RmP can derange this process by impeding removal of
all-trans-retinal from the disc. In a recessive form of macular
degeneration called Stargardt's disease ( 1/10,000 incidence rate
often affecting children; 25,000 affected individuals in the U.S.),
the gene encoding RmP, abcr, is mutated, and the transporter is
nonfunctional. As a result, all-trans-retinal and/or
N-retinylidene-PE become trapped in the disc. The N-retinylidene-PE
can then react with another molecule of all-trans-retinal to form
N-retinylidene-N-retinylethanolamine (A.sub.2E); this is summarized
in FIG. 2. As noted above, some A.sub.2E is formed even under
normal conditions; however, its production is greatly increased
when its precursors accumulate inside the discs due to the
defective transporter, and can thereby cause macular
degeneration.
[0006] Other forms of macular degeneration may also result from
pathologies that result in lipofuscin accumulation. A dominant form
of Stargardt's disease, known as chromosome 6-linked autosomal
dominant macular dystrophy (ADMD, OMIM #600110), is caused by a
mutation in the gene encoding elongation of very long chain fatty
acids-4, elov14.
[0007] There are few, if any, preventative treatments for AMD, and
therapeutic interventions are available for only certain, less
common, forms of the disease.
SUMMARY
[0008] This disclosure relates to compositions, systems, and
methods for managing macular degeneration, and, more specifically,
for preventing the accumulation of retinotoxic compounds in and
around the retinal pigment epithelium.
[0009] In one embodiment, the accumulation of A.sub.2E in rod
outer-segment discs is prevented or reduced. It has been found that
A.sub.2E production in discs can be reduced by administering a drug
that limits the visual cycle. The limitation can be achieved in a
number of ways. In one approach, a drug can effectively
short-circuit the portion of the visual cycle that generates the
A.sub.2E precursor, all-trans-retinal. In another approach, a drug
can inhibit particular steps in the visual cycle necessary for
synthesizing all-trans-retinal. In yet another approach, a drug can
prevent binding of intermediate products (retinyl esters) to
certain chaperone proteins in the retinal pigment epithelium.
[0010] In one embodiment, a method of treating or preventing
macular degeneration in a subject may include administering to the
subject a drug that short-circuits the visual cycle at a step of
the visual cycle that occurs outside a disc of a rod photoreceptor
cell. In another embodiment, a method of treating or preventing
macular degeneration in a subject may include administering to the
subject a drug that inhibits and/or interferes with at least one of
lecithin retinol acyl transferase, RPE65, 11-cis-retinol
dehydrogenase, and isomerohydrolase.
[0011] In yet another embodiment, a method of identifying a macular
degeneration drug may include administering a candidate drug to a
subject having, or at risk for developing, macular degeneration,
and measuring accumulation of a retinotoxic compound in the retinal
pigment epithelium of the subject.
[0012] A wide variety of drugs are contemplated for use. In some
embodiments, inhibitors of the visual cycle include retinoic acid
analogs. In other embodiments, drugs that short circuit the visual
cycle include aromatic amines and hydrazines.
BRIEF DESCRIPTION OF THE FIGURES
[0013] FIG. 1 depicts the visual cycle.
[0014] FIG. 2 depicts the synthesis of A.sub.2E.
[0015] FIG. 3 depicts an intervention for short-circuiting the
visual cycle.
[0016] FIGS. 4A-B show that TDT (A) and TDH (B) bind to mRPE65 with
high affinities.
[0017] FIGS. 5A-B show ERG-b effects of TDT and TDH. FIG. 5A shows
acute effects of drugs 1 h after 50 mg/kg (i.p). FIG. 5B shows
persistent effects of TDT and TDH 3 days after treatment.
[0018] FIGS. 6A-C show that certain isoprenoid mRPE65 antagonists
inhibit 11-cis-retinal regeneration.
[0019] FIGS. 7A-D shows that certain isoprenoid mRPE65 antagonists
lower A.sub.2E accumulation.
DETAILED DESCRIPTION
Overview
[0020] The present disclosure provides compositions and methods for
managing macular degeneration by preventing or reducing the
accumulation of A.sub.2E in rod outer-segment discs. A.sub.2E
accumulation can be prevented or reduced by decreasing the amount
of all-trans-retinal present in discs of rod outer segments. In one
approach, a drug may be administered that inhibits one or more
enzymatic steps in the visual cycle, so that production of
all-trans-retinal is diminished. In another approach, a drug may be
administered that drives the isomerization of 11-cis-retinal to
all-trans-retinal in the RPE, thereby decreasing the amount
11-cis-retinal that returns to the outer segment discs to be
re-isomerized to all-trans-retinal.
Definitions
[0021] For convenience, before further description of exemplary
embodiments, certain terms employed in the specification, examples,
and appended claims are collected here. These definitions should be
read in light of the remainder of the disclosure and as understood
by a person of skill in the art.
[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 term "access device" is an art-recognized term and
includes any medical device adapted for gaining or maintaining
access to an anatomic area. Such devices are familiar to artisans
in the medical and surgical fields. An access device may be a
needle, a catheter, a cannula, a trocar, a tubing, a shunt, a
drain, or an endoscope such as an otoscope, nasopharyngoscope,
bronchoscope, or any other endoscope adapted for use in the joint
area, or any other medical device suitable for entering or
remaining positioned within the preselected anatomic area.
[0024] The terms "biocompatible compound" and "biocompatibility"
when used in relation to compounds are art-recognized. For example,
biocompatible compounds include compounds that are neither
themselves toxic to the host (e.g., an animal or human), nor
degrade (if the compound degrades) at a rate that produces
monomeric or oligomeric subunits or other byproducts at toxic
concentrations in the host. In certain embodiments, biodegradation
generally involves degradation of the compound in an organism,
e.g., into its monomeric subunits, which may be known to be
effectively non-toxic. Intermediate oligomeric products resulting
from such degradation may have different toxicological properties,
however, or biodegradation may involve oxidation or other
biochemical reactions that generate molecules other than monomeric
subunits of the compound. Consequently, in certain embodiments,
toxicology of a biodegradable compound intended for in vivo use,
such as implantation or injection into a patient, may be determined
after one or more toxicity analyses. It is not necessary that any
subject composition have a purity of 100% to be deemed
biocompatible; indeed, it is only necessary that the subject
compositions be biocompatible as set forth above. Hence, a subject
composition may comprise compounds comprising 99%, 98%, 97%, 96%,
95%, 90%, 85%, 80%, 75% or even less of biocompatible compounds,
e.g., including compounds and other materials and excipients
described herein, and still be biocompatible.
[0025] To determine whether a compound or other material is
biocompatible, it may be necessary to conduct a toxicity analysis.
Such assays are well known in the art. One example of such an assay
may be performed with live carcinoma cells, such as GT3TKB tumor
cells, in the following manner: the sample is degraded in 1M NaOH
at 37.degree. C. until complete degradation is observed. The
solution is then neutralized with 1M HCl. About 200 .mu.L of
various concentrations of the degraded sample products are placed
in 96-well tissue culture plates and seeded with human gastric
carcinoma cells (GT3TKB) at 10.sup.4/well density. The degraded
sample products are incubated with the GT3TKB cells for 48 hours.
The results of the assay may be plotted as % relative growth vs.
concentration of degraded sample in the tissue-culture well. In
addition, compounds and formulations may also be evaluated by
well-known in vivo tests, such as subcutaneous implantations in
rats to confirm that they do not cause significant levels of
irritation or inflammation at the subcutaneous implantation
sites.
[0026] The term "biodegradable" is art-recognized, and includes
compounds, compositions and formulations, such as those described
herein, that are intended to degrade during use. Biodegradable
compounds typically differ from non-biodegradable compounds in that
the former may be degraded during use. In certain embodiments, such
use involves in vivo use, such as in vivo therapy, and in other
certain embodiments, such use involves in vitro use. In general,
degradation attributable to biodegradability involves the
degradation of a biodegradable compound into its component
subunits, or digestion, e.g., by a biochemical process, of the
compound into smaller subunits. In certain embodiments, two
different types of biodegradation may generally be identified. For
example, one type of biodegradation may involve cleavage of bonds
(whether covalent or otherwise) in the compound. In such
biodegradation, monomers and oligomers typically result, and even
more typically, such biodegradation occurs by cleavage of a bond
connecting one or more of substituents of a compound. In contrast,
another type of biodegradation may involve cleavage of a bond
(whether covalent or otherwise) internal to side chain or that
connects a side chain to the compound. For example, a therapeutic
agent or other chemical moiety attached as a side chain to the
compound may be released by biodegradation. In certain embodiments,
one or the other or both generally types of biodegradation may
occur during use of a compound. As used herein, the term
"biodegradation" encompasses both general types of
biodegradation.
[0027] The degradation rate of a biodegradable compound often
depends in part on a variety of factors, including the chemical
identity of the linkage responsible for any degradation, the
molecular weight, crystallinity, biostability, and degree of
cross-linking of such compound, the physical characteristics of the
implant, shape and size, and the mode and location of
administration. For example, the greater the molecular weight, the
higher the degree of crystallinity, and/or the greater the
biostability, the biodegradation of any biodegradable compound is
usually slower. The term "biodegradable" is intended to cover
materials and processes also termed "bioerodible".
[0028] In certain embodiments, if the biodegradable compound also
has a therapeutic agent or other material associated with it, the
biodegradation rate of such compound may be characterized by a
release rate of such materials. In such circumstances, the
biodegradation rate may depend on not only the chemical identity
and physical characteristics of the compound, but also on the
identity of any such material incorporated therein.
[0029] In certain embodiments, compound formulations bio degrade
within a period that is acceptable in the desired application. In
certain embodiments, such as in vivo therapy, such degradation
occurs in a period usually less than about five years, one year,
six months, three months, one month, fifteen days, five days, three
days, or even one day on exposure to a physiological solution with
a pH between 6 and 8 having a temperature of between 25 and
37.degree. C. In other embodiments, the compound degrades in a
period of between about one hour and several weeks, depending on
the desired application.
[0030] 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.
[0031] The term "drug delivery device" is an art-recognized term
and refers to any medical device suitable for the application of a
drug to a targeted organ or anatomic region. The term includes
those devices that transport or accomplish the instillation of the
compositions towards the targeted organ or anatomic area, even if
the device itself is not formulated to include the composition. As
an example, a needle or a catheter through which the composition is
inserted into an anatomic area or into a blood vessel or other
structure related to the anatomic area is understood to be a drug
delivery device. As a further example, a stent or a shunt or a
catheter that has the composition included in its substance or
coated on its surface is understood to be a drug delivery
device.
[0032] When used with respect to a therapeutic agent or other
material, the term "sustained release" is art-recognized. For
example, a subject composition that releases a substance over time
may exhibit sustained release characteristics, in contrast to a
bolus type administration in which the entire amount of the
substance is made biologically available at one time. For example,
in particular embodiments, upon contact with body fluids including
blood, tissue fluid, lymph or the like, the compound matrices
(formulated as provided herein and otherwise as known to one of
skill in the art) may undergo gradual degradation (e.g., through
hydrolysis) with concomitant release of any material incorporated
therein, for a sustained or extended period (as compared to the
release from a bolus). This release may result in prolonged
delivery of therapeutically effective amounts of any incorporated a
therapeutic agent. Sustained release will vary in certain
embodiments as described in greater detail below.
[0033] The term "delivery agent" is an art-recognized term, and
includes molecules that facilitate the intracellular delivery of a
therapeutic agent or other material. Examples of delivery agents
include: sterols (e.g., cholesterol) and lipids (e.g., a cationic
lipid, virosome or liposome).
[0034] The term "or" as used herein should be understood to mean
"and/or", unless the context clearly indicates otherwise.
[0035] The phrases "parenteral administration" and "administered
parenterally" are art-recognized terms, and include 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 intrasternal injection and
infusion.
[0036] The term "treating" is art-recognized and includes
inhibiting a disease, disorder or condition in a subject having
been diagnosed with the disease, disorder, or condition, 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.
[0037] The term "preventing" is art-recognized and includes
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.
[0038] The term "fluid" is art-recognized to refer to a non-solid
state of matter in which the atoms or molecules are free to move in
relation to each other, as in a gas or liquid. If unconstrained
upon application, a fluid material may flow to assume the shape of
the space available to it, covering for example, the surfaces of an
excisional site or the dead space left under a flap. A fluid
material may be inserted or injected into a limited portion of a
space and then may flow to enter a larger portion of the space or
its entirety. Such a material may be termed "flowable." This term
is art-recognized and includes, for example, liquid compositions
that are capable of being sprayed into a site; injected with a
manually operated syringe fitted with, for example, a 23-gauge
needle; or delivered through a catheter. Also included in the term
"flowable" are those highly viscous, "gel-like" materials at room
temperature that may be delivered to the desired site by pouring,
squeezing from a tube, or being injected with any one of the
commercially available injection devices that provide injection
pressures sufficient to propel highly viscous materials through a
delivery system such as a needle or a catheter. When the compound
used is itself flowable, a composition comprising it need not
include a biocompatible solvent to allow its dispersion within a
body cavity. Rather, the flowable compound may be delivered into
the body cavity using a delivery system that relies upon the native
flowability of the material for its application to the desired
tissue surfaces. For example, if flowable, a composition comprising
compounds can be injected to form, after injection, a temporary
biomechanical barrier to coat or encapsulate internal organs or
tissues, or it can be used to produce coatings for solid
implantable devices. In certain instances, flowable subject
compositions have the ability to assume, over time, the shape of
the space containing it at body temperature.
[0039] Viscosity is understood herein as it is recognized in the
art to be the internal friction of a fluid or the resistance to
flow exhibited by a fluid material when subjected to deformation.
The degree of viscosity of the compound may be adjusted by the
molecular weight of the compound and other methods for altering the
physical characteristics of a specific compound will be evident to
practitioners of ordinary skill with no more than routine
experimentation. The molecular weight of the compound used may vary
widely, depending on whether a rigid solid state (higher molecular
weights) desirable, or whether a fluid state (lower molecular
weights) is desired.
[0040] The phrase "pharmaceutically acceptable" is art-recognized.
In certain embodiments, the term includes compositions, compounds
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.
[0041] The phrase "pharmaceutically acceptable carrier" is
art-recognized, and includes, for example, 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.
[0042] The term "pharmaceutically acceptable salts" is
art-recognized, and includes relatively non-toxic, inorganic and
organic acid addition salts of compositions, including without
limitation, therapeutic agents, excipients, other materials and the
like. Examples of pharmaceutically acceptable salts include those
derived from mineral acids, such as hydrochloric acid and sulfuric
acid, and those derived from organic acids, such as ethanesulfonic
acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like.
Examples of suitable inorganic bases for the formation of salts
include the hydroxides, carbonates, and bicarbonates of ammonia,
sodium, lithium, potassium, calcium, magnesium, aluminum, zinc and
the like. Salts may also be formed with suitable organic bases,
including those that are non-toxic and strong enough to form such
salts. For purposes of illustration, the class of such organic
bases may include mono-, di-, and trialkylamines, such as
methylamine, dimethylamine, and triethylamine; mono-, di- or
trihydroxyalkylamines such as mono-, di-, and triethanolamine;
amino acids, such as arginine and lysine; guanidine;
N-methylglucosamine; N-methylglucamine; L-glutamine;
N-methylpiperazine; morpholine; ethylenediamine;
N-benzylphenethylamine; (trihydroxymethyl)aminoethane; and the
like. See, for example, J. Pharm. Sci., 66:1-19 (1977).
[0043] 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.
[0044] The term "prophylactic or therapeutic" treatment is
art-recognized and includes 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).
[0045] The terms "therapeutic agent", "drug", "medicament" and
"bioactive substance" are art-recognized and include 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 macular degeneration. The terms include without
limitation pharmaceutically acceptable salts thereof and pro-drugs.
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.
[0046] 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
compound, 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 (e.g., prevent the spread of) a tumor
or other 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 compound matrix,
which will depend in part on the chemical and physical
characteristics of the compound; the identity of the agent; the
mode and method of administration; and any other materials
incorporated in the compound matrix in addition to the agent.
[0047] "Radiosensitizer" is defined as a therapeutic agent that,
upon administration in a therapeutically effective amount, promotes
the treatment of one or more diseases or conditions that are
treatable with electromagnetic radiation. In general,
radiosensitizers are intended to be used in conjunction with
electromagnetic radiation as part of a prophylactic or therapeutic
treatment. Appropriate radiosensitizers to use in conjunction with
treatment with the subject compositions will be known to those of
skill in the art.
[0048] "Electromagnetic radiation" as used in this specification
includes, but is not limited to, radiation having the wavelength of
10-20 to 10 meters. Particular embodiments of electromagnetic
radiation employ the electromagnetic radiation of: gamma-radiation
(10.sup.-20 to 10.sup.-13 m), x-ray radiation (10.sup.-11 to
10.sup.-9 m), ultraviolet light (10 nm to 400 nm), visible light
(400 nm to 700 nm), infrared radiation (700 nm to 1.0 mm), and
microwave radiation (1 mm to 30 cm).
[0049] "Retinol binding protein" (RBP) is the principal carrier of
all-trans-retinol, which comprises over 90% of serum vitamin A. RBP
is found in serum in association with a cotransport protein called
transthyretin or prealbumin. Within cells, retinol and its
metabolites are bound to retinol-binding proteins in the cytosol
and nucleus (Folli et al. J. Biol. Chem., Vol. 277:41970; Ong, et
al. (1994) in The Retinoids: Biology, Chemistry and Medicine
(Sporn, M. B., Roberts, A. B., and Goodman, D. S., eds), pp.
283-318, Raven Press Ltd., New York; and Li et al. (1996) Annu.
Rev. Nutr. 16, 205). The eye shows a very marked preference for
acquiring retinol from the retinol-RBP complex (Vogel et al. (2002)
Biochemistry. 41(51):15360). The RBP family contains 7 members:
RBP1-7. RBP1 is also referred to as cellular RBP1, HGNC:9919,
CRABP-I, CRBP, CRBP1, and RBPC, and its nucleotide and amino acid
sequences are set forth in GenBank Accession Nos. NM.sub.--002899
and NP.sub.--002890. RBP2 is also referred to as cellular RBP2,
HGNC:9920, CRABP-II, CRBP2, CRBPII, and RBPC2, and its nucleotide
and amino acid sequences are set forth in GenBank Accession Nos.
NM.sub.--004164 and NP.sub.--004155. RBP3 is also referred to as
insterstitial RBP3, IRBP; RBPI; D10S64; D10S65; and D10S66, and its
nucleotide and amino acid sequences are set forth in GenBank
Accession Nos. NM.sub.--002900 and NP.sub.--002891. RBP4 is also
referred to as plasma RBP4, and its nucleotide and amino acid
sequences are set forth in GenBank Accession Nos. NM.sub.--006744
and NP.sub.--006735. RBP5 is also referred to as cellular RBP5,
CRBP3; CRBPIII; and CRBP-III, and its nucleotide and amino acid
sequences are set forth in GenBank Accession Nos. NM.sub.--031491
and NP.sub.--113679. RBP6 is also referred to as cellular retinoic
acid-binding protein 2, cellular retinoic acid binding protein 2,
CRABP2, HGNC:2339, CRABP-II, and its nucleotide and amino acid
sequences are set forth in GenBank Accession Nos. NM.sub.--001878
and NP.sub.--001869. RBP7 is also referred to as cellular RBP7,
HGNC:30316, CRBP4, CRBPIV, and MGC70641, and its nucleotide and
amino acid sequences are set forth in GenBank Accession Nos:
NM.sub.--052960 and NP.sub.--443192. The RBP that is believed to
import Vitamin A into the eye are the CRBPs. Inhibition of other
RBPs may also be used for treating and/or preventing diseases of
the eye.
[0050] The phrases "systemic administration," "administered
systemically," "peripheral administration" and "administered
peripherally" are art-recognized, and include the administration of
a subject composition or other material at a site remote from the
site affected by the disease being treated. Administration of an
agent directly into, onto or in the vicinity of a lesion of the
disease being treated, even if the agent is subsequently
distributed systemically, may be termed "local" or "topical" or
"regional" administration. The term "ED.sub.50" is art-recognized.
In certain embodiments, ED.sub.50 means 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 "LD.sub.50" is art-recognized.
In certain embodiments, LD.sub.50 means the dose of a drug which is
lethal in 50% of test subjects. The term "therapeutic index" is an
art-recognized term which refers to the therapeutic index of a
drug, defined as LD.sub.50/ED.sub.50.
[0051] The terms "incorporated" and "encapsulated" are
art-recognized when used in reference to a therapeutic agent and a
compound, such as a composition disclosed herein. In certain
embodiments, these terms include incorporating, formulating or
otherwise including such agent into a composition which allows for
sustained release of such agent in the desired application. The
terms may contemplate any manner by which a therapeutic agent or
other material is incorporated into a compound matrix, including
for example: the compound is a polymer, and the agent is attached
to a monomer of such polymer (by covalent or other binding
interaction) and having such monomer be part of the polymerization
to give a polymeric formulation, distributed throughout the
polymeric matrix, appended to the surface of the polymeric matrix
(by covalent or other binding interactions), encapsulated inside
the polymeric matrix, etc. The term "co-incorporation" or
"co-encapsulation" refers to the incorporation of a therapeutic
agent or other material and at least one other a therapeutic agent
or other material in a subject composition.
[0052] More specifically, the physical form in which a therapeutic
agent or other material is encapsulated in compounds may vary with
the particular embodiment. For example, a therapeutic agent or
other material may be first encapsulated in a microsphere and then
combined with the compound in such a way that at least a portion of
the microsphere structure is maintained. Alternatively, a
therapeutic agent or other material may be sufficiently immiscible
in a controlled-release compound that it is dispersed as small
droplets, rather than being dissolved, in the compound. Any form of
encapsulation or incorporation is contemplated by the present
disclosure, in so much as the sustained release of any encapsulated
therapeutic agent or other material determines whether the form of
encapsulation is sufficiently acceptable for any particular
use.
[0053] The term "biocompatible plasticizer" is art-recognized, and
includes materials which are soluble or dispersible in the
controlled-release compositions described herein, which increase
the flexibility of the compound matrix, and which, in the amounts
employed, are biocompatible. Suitable plasticizers are well known
in the art and include those disclosed in U.S. Pat. Nos. 2,784,127
and 4,444,933. Specific plasticizers include, by way of example,
acetyl tri-n-butyl citrate (about 20 weight percent or less),
acetyl trihexyl citrate (about 20 weight percent or less), butyl
benzyl phthalate, dibutyl phthalate, dioctylphthalate, n-butyryl
tri-n-hexyl citrate, diethylene glycol dibenzoate (c. 20 weight
percent or less) and the like.
[0054] "Small molecule" is an art-recognized term. In certain
embodiments, this term 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.
[0055] The term "alkyl" is art-recognized, and includes saturated
aliphatic groups, including straight-chain alkyl groups,
branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl
substituted cycloalkyl groups, and cycloalkyl substituted alkyl
groups. In certain embodiments, a straight chain or branched chain
alkyl has about 30 or fewer carbon atoms in its backbone (e.g.,
C1-C30 for straight chain, C3-C30 for branched chain), and
alternatively, about 20 or fewer. Likewise, cycloalkyls have from
about 3 to about 10 carbon atoms in their ring structure, and
alternatively about 5, 6 or 7 carbons in the ring structure.
[0056] Unless the number of carbons is otherwise specified, "lower
alkyl" refers to an alkyl group, as defined above, but having from
one to about ten carbons, alternatively from one to about six
carbon atoms in its backbone structure. Likewise, "lower alkenyl"
and "lower alkynyl" have similar chain lengths.
[0057] The term "aralkyl" is art-recognized and refers to an alkyl
group substituted with an aryl group (e.g., an aromatic or
heteroaromatic group).
[0058] The terms "alkenyl" and "alkynyl" are art-recognized and
refer to unsaturated aliphatic groups analogous in length and
possible substitution to the alkyls described above, but that
contain at least one double or triple bond respectively.
[0059] The term "aryl" is art-recognized and refers to 5-, 6- and
7-membered single-ring aromatic groups that may include from zero
to four heteroatoms, for example, benzene, naphthalene, anthracene,
pyrene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole,
triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine,
and the like. Those aryl groups having heteroatoms in the ring
structure may also be referred to as "aryl heterocycles" or
"heteroaromatics." The aromatic ring may be substituted at one or
more ring positions with such substituents as described above, for
example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl,
cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino,
amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether,
alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester,
heterocyclyl, aromatic or heteroaromatic moieties, --CF.sub.3,
--CN, or the like. The term "aryl" also includes polycyclic ring
systems having two or more cyclic rings in which two or more
carbons are common to two adjoining rings (the rings are "fused
rings") wherein at least one of the rings is aromatic, e.g., the
other cyclic rings may be cycloalkyls, cycloalkenyls,
cycloalkynyls, aryls and/or heterocyclyls.
[0060] The terms ortho, meta and para are art-recognized and refer
to 1,2-, 1,3- and 1,4-disubstituted benzenes, respectively. For
example, the names 1,2-dimethylbenzene and ortho-dimethylbenzene
are synonymous.
[0061] The terms "heterocyclyl", "heteroaryl", or "heterocyclic
group" are art-recognized and refer to 3- to about 10-membered ring
structures, alternatively 3- to about 7-membered rings, whose ring
structures include one to four heteroatoms. Heterocycles may also
be polycycles. Heterocyclyl groups include, for example, thiophene,
thianthrene, furan, pyran, isobenzofuran, chromene, xanthene,
phenoxanthene, pyrrole, imidazole, pyrazole, isothiazole,
isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine,
isoindole, indole, indazole, purine, quinolizine, isoquinoline,
quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline,
cinnoline, pteridine, carbazole, carboline, phenanthridine,
acridine, pyrimidine, phenanthroline, phenazine, phenarsazine,
phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane,
thiolane, oxazole, piperidine, piperazine, morpholine, lactones,
lactams such as azetidinones and pyrrolidinones, sultams, sultones,
and the like. The heterocyclic ring may be substituted at one or
more positions with such substituents as described above, as for
example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,
hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate,
phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,
ketone, aldehyde, ester, a heterocyclyl, an aromatic or
heteroaromatic moiety, --CF.sub.3, --CN, or the like.
[0062] The terms "polycyclyl" or "polycyclic group" are
art-recognized and refer to two or more rings (e.g., cycloalkyls,
cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) in which
two or more carbons are common to two adjoining rings, e.g., the
rings are "fused rings". Rings that are joined through non-adjacent
atoms are termed "bridged" rings. Each of the rings of the
polycycle may be substituted with such substituents as described
above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl,
cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido,
phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether,
alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an
aromatic or heteroaromatic moiety, --CF.sub.3, --CN, or the
like.
[0063] The term "carbocycle" is art-recognized and refers to an
aromatic or non-aromatic ring in which each atom of the ring is
carbon.
[0064] The term "nitro" is art-recognized and refers to --NO.sub.2;
the term "halogen" is art-recognized and refers to --F, --Cl, --Br
or --I; the term "sulfhydryl" is art-recognized and refers to SH;
the term "hydroxyl" means --OH; and the term "sulfonyl" is
art-recognized and refers to SO.sub.2--. "Halide" designates the
corresponding anion of the halogens, and "pseudohalide" has the
definition set forth on page 560 of "Advanced Inorganic Chemistry"
by Cotton and Wilkinson.
[0065] The terms "amine" and "amino" are art-recognized and refer
to both unsubstituted and substituted amines, e.g., a moiety that
may be represented by the general formulas:
##STR00001##
wherein R50, R51 and R52 each independently represent a hydrogen,
an alkyl, an alkenyl, (CH.sub.2).sub.m--R61, or R50 and R51, taken
together with the N atom to which they are attached complete a
heterocycle having from 4 to 8 atoms in the ring structure; R61
represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle or
a polycycle; and m is zero or an integer in the range of 1 to 8. In
other embodiments, R50 and R51 (and optionally R52) each
independently represent a hydrogen, an alkyl, an alkenyl, or
--(CH.sub.2).sub.m--R61. Thus, the term "alkylamine" includes an
amine group, as defined above, having a substituted or
unsubstituted alkyl attached thereto, i.e., at least one of R50 and
R51 is an alkyl group.
[0066] The term "acylamino" is art-recognized and refers to a
moiety that may be represented by the general formula:
##STR00002##
wherein R50 is as defined above, and R54 represents a hydrogen, an
alkyl, an alkenyl or --(CH.sub.2).sub.m--R61, where m and R61 are
as defined above.
[0067] The term "amido" is art recognized as an amino-substituted
carbonyl and includes a moiety that may be represented by the
general formula:
##STR00003##
wherein R50 and R51 are as defined above. Certain embodiments of
the amide in the present invention will not include imides which
may be unstable.
[0068] The term "alkylthio" refers to an alkyl group, as defined
above, having a sulfur radical attached thereto. In certain
embodiments, the "alkylthio" moiety is represented by one of S
alkyl, --S-alkenyl, --S-alkynyl, and --S--(CH.sub.2).sub.m--R61,
wherein m and R61 are defined above. Representative alkylthio
groups include methylthio, ethyl thio, and the like.
The term "carboxyl" is art recognized and includes such moieties as
may be represented by the general formulas:
##STR00004##
wherein X50 is a bond or represents an oxygen or a sulfur, and R55
and R56 represents a hydrogen, an alkyl, an alkenyl,
--(CH.sub.2).sub.m--R61 or a pharmaceutically acceptable salt, R56
represents a hydrogen, an alkyl, an alkenyl or
--(CH.sub.2).sub.m--R61, where m and R61 are defined above. Where
X50 is an oxygen and R55 or R56 is not hydrogen, the formula
represents an "ester". Where X50 is an oxygen, and R55 is as
defined above, the moiety is referred to herein as a carboxyl
group, and particularly when R55 is a hydrogen, the formula
represents a "carboxylic acid". Where X50 is an oxygen, and R56 is
hydrogen, the formula represents a "formate". In general, where the
oxygen atom of the above formula is replaced by sulfur, the formula
represents a "thiolcarbonyl" group. Where X50 is a sulfur and R55
or R56 is not hydrogen, the formula represents a "thiolester."
Where X50 is a sulfur and R55 is hydrogen, the formula represents a
"thiolcarboxylic acid." Where X50 is a sulfur and R56 is hydrogen,
the formula represents a "thiolformate." On the other hand, where
X50 is a bond, and R55 is not hydrogen, the above formula
represents a "ketone" group. Where X50 is a bond, and R55 is
hydrogen, the above formula represents an "aldehyde" group.
[0069] The term "carbamoyl" refers to --O(C.dbd.O)NRR', where R and
R' are independently H, aliphatic groups, aryl groups or heteroaryl
groups.
[0070] The term "oxo" refers to a carbonyl oxygen (.dbd.O).
[0071] The terms "oxime" and "oxime ether" are art-recognized and
refer to moieties that may be represented by the general
formula:
##STR00005##
wherein R75 is hydrogen, allyl, cycloalkyl, alkenyl, alkynyl, aryl,
aralkyl, or --(CH.sub.2).sub.m--R61. The moiety is an "oxime" when
R is H; and it is an "oxime ether" when R is alkyl, cycloalkyl,
alkenyl, alkynyl, aryl, aralkyl, or --(CH.sub.2).sub.m--R61.
[0072] The terms "alkoxyl" or "alkoxy" are art-recognized and refer
to an alkyl group, as defined above, having an oxygen radical
attached thereto. Representative alkoxyl groups include methoxy,
ethoxy, propyloxy, tert-butoxy and the like. An "ether" is two
hydrocarbons covalently linked by an oxygen. Accordingly, the
substituent of an alkyl that renders that alkyl an ether is or
resembles an alkoxyl, such as may be represented by one of
--O-alkyl, --O-alkenyl, O-alkynyl, --O--(CH.sub.2).sub.m--R61,
where m and R61 are described above.
[0073] The term "sulfonate" is art recognized and refers to a
moiety that may be represented by the general formula:
##STR00006##
in which R57 is an electron pair, hydrogen, alkyl, cycloalkyl, or
aryl.
[0074] The term "sulfate" is art recognized and includes a moiety
that may be represented by the general formula:
##STR00007##
in which R57 is as defined above.
[0075] The term "sulfonamido" is art recognized and includes a
moiety that may be represented by the general formula:
##STR00008##
in which R50 and R56 are as defined above.
[0076] The term "sulfamoyl" is art-recognized and refers to a
moiety that may be represented by the general formula:
##STR00009##
in which R50 and R51 are as defined above.
[0077] The term "sulfonyl" is art-recognized and refers to a moiety
that may be represented by the general formula:
##STR00010##
in which R58 is one of the following: hydrogen, alkyl, alkenyl,
alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl.
[0078] The term "sulfoxido" is art-recognized and refers to a
moiety that may be represented by the general formula:
##STR00011##
in which R58 is defined above.
[0079] The term "phosphoryl" is art-recognized and may in general
be represented by the formula:
##STR00012##
wherein Q50 represents S or O, and R59 represents hydrogen, a lower
alkyl or an aryl. When used to substitute, e.g., an alkyl, the
phosphoryl group of the phosphorylalkyl may be represented by the
general formulas:
##STR00013##
wherein Q50 and R59, each independently, are defined above, and Q51
represents O, S or N. When Q50 is S, the phosphoryl moiety is a
"phosphorothioate".
[0080] The term "phosphoramidite" is art-recognized and may be
represented in the general formulas:
##STR00014##
wherein Q51, R50, R51 and R59 are as defined above. The term
"phosphonamidite" is art-recognized and may be represented in the
general formulas:
##STR00015##
wherein Q51, R50, R51 and R59 are as defined above, and R60
represents a lower alkyl or an aryl.
[0081] Analogous substitutions may be made to alkenyl and alkynyl
groups to produce, for example, aminoalkenyls, aminoalkynyls,
amidoalkenyls, amidoalkynyls, iminoalkenyls, iminoalkynyls,
thioalkenyls, thioalkynyls, carbonyl-substituted alkenyls or
alkynyls.
The definition of each expression, e.g. alkyl, m, n, and the like,
when it occurs more than once in any structure, is intended to be
independent of its definition elsewhere in the same structure.
[0082] The term "selenoalkyl" is art-recognized and refers to an
alkyl group having a substituted seleno group attached thereto.
Exemplary "selenoethers" which may be substituted on the alkyl are
selected from one of -Se-alkyl, -Se-alkenyl, -Se-alkynyl, and
-Se-(CH.sub.2).sub.m--R61, m and R61 being defined above.
[0083] The terms triflyl, tosyl, mesyl, and nonaflyl are
art-recognized and refer to trifluoromethanesulfonyl,
p-toluenesulfonyl, methanesulfonyl, and nonafluorobutanesulfonyl
groups, respectively. The terms triflate, tosylate, mesylate, and
nonaflate are art-recognized and refer to trifluoromethanesulfonate
ester, p-toluenesulfonate ester, methanesulfonate ester, and
nonafluorobutanesulfonate ester functional groups and molecules
that contain said groups, respectively.
[0084] The abbreviations Me, Et, Ph, Tf, Nf, Ts, and Ms represent
methyl, ethyl, phenyl, trifluoromethanesulfonyl,
nonafluorobutanesulfonyl, p-toluenesulfonyl and methanesulfonyl,
respectively. A more comprehensive list of the abbreviations
utilized by organic chemists of ordinary skill in the art appears
in the first issue of each volume of the Journal of Organic
Chemistry; this list is typically presented in a table entitled
Standard List of Abbreviations.
[0085] Certain compounds contained in compositions of the present
invention may exist in particular geometric or stereoisomeric
forms. In addition, polymers of the present invention may also be
optically active. The present invention contemplates all such
compounds, including cis- and trans-isomers, R- and S-enantiomers,
diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures
thereof, and other mixtures thereof, as falling within the scope of
the invention. Additional asymmetric carbon atoms may be present in
a substituent such as an alkyl group. All such isomers, as well as
mixtures thereof, are intended to be included in this
invention.
[0086] If, for instance, a particular enantiomer of compound of the
present invention is desired, it may be prepared by asymmetric
synthesis, or by derivation with a chiral auxiliary, where the
resulting diastereomeric mixture is separated and the auxiliary
group cleaved to provide the pure desired enantiomers.
Alternatively, where the molecule contains a basic functional
group, such as amino, or an acidic functional group, such as
carboxyl, diastereomeric salts are formed with an appropriate
optically-active acid or base, followed by resolution of the
diastereomers thus formed by fractional crystallization or
chromatographic means well known in the art, and subsequent
recovery of the pure enantiomers.
[0087] It will be understood that "substitution" or "substituted
with" includes the implicit proviso that such substitution is in
accordance with permitted valence of the substituted atom and the
substituent, and that the substitution results in a stable
compound, e.g., which does not spontaneously undergo transformation
such as by rearrangement, cyclization, elimination, or other
reaction.
[0088] The term "substituted" is also contemplated to include all
permissible substituents of organic compounds. In a broad aspect,
the permissible substituents include acyclic and cyclic, branched
and unbranched, carbocyclic and heterocyclic, aromatic and
nonaromatic substituents of organic compounds. Illustrative
substituents include, for example, those described herein above.
The permissible substituents may be one or more and the same or
different for appropriate organic compounds. For purposes of this
invention, the heteroatoms such as nitrogen may have hydrogen
substituents and/or any permissible substituents of organic
compounds described herein which satisfy the valences of the
heteroatoms. This invention is not intended to be limited in any
manner by the permissible substituents of organic compounds.
[0089] 3. Compositions
[0090] As described above, macular degeneration may be treated or
prevented by interfering with the visual cycle in such a way that
diminishes the amount of all-trans-retinal present in the discs of
the rod photoreceptor outer segments. Production of retinotoxic
compounds by cone cells is negligible and may be ignored, because
rods represent 95% of all photoreceptors.
[0091] FIG. 1 depicts the mammalian visual cycle. In the course of
the visual cycle, a complex of 11-cis-retinal and opsin, known as
rhodopsin, passes through a series of biochemical steps initiated
by the absorption of light. Various steps of this cycle in distinct
places. As FIG. 1 illustrates, the initial steps of light
absorption to the dissociation of opsin and the formation of
all-trans-retinal occur in the discs of the rod photoreceptor cell
outer segment. The reduction of all-trans-retinal to
all-trans-retinol takes place in the cytoplasm of the rod cell, and
the remaining steps to regenerate 11-cis-retinal occur in the
retinal pigment epithelium (RPE).
[0092] At least two broad approaches are contemplated for
preventing the accumulation of all-trans-retinal in the disc. In
one approach, one or more enzymatic steps or chaperone binding
steps in the visual cycle may be inhibited so that the synthetic
pathway to all-trans-retinal is blocked. In another approach, a
portion of the visual cycle is "short-circuited," i.e., an early
intermediate in the cycle is shunted to an intermediate that is two
or more steps later in the visual cycle, so that these steps of the
cycle are bypassed while the all-trans-retinal precursors are not
in the disc.
[0093] A. Enzyme Inhibitors
[0094] Limiting the flux of retinoids through the visual cycle can
be achieved by inhibiting any of the key biochemical reactions of
the visual cycle. Each step of the cycle is potentially addressable
in this fashion. Inhibiting an enzymatic step could thus be used to
"stall" the visual cycle in the RPE, thereby keeping
all-tran's-retinal out of the discs.
[0095] Other steps in the visual cycle are also prone to
inhibition. For example, as shown in FIG. 1, several enzymes act
upon all-trans-retinol and its derivatives upon its return to the
RPE, including LRAT (lecithin retinol acyl transferase),
11-cis-retinol dehydrogenase and IMH (isomerohydrolase). In
addition, the chaperone RPE65 binds retinyl esters to make those
typically hydrophobic compounds available to IMH for processing to
11-cis-retinol. These enzymes and chaperone may be targeted for
inhibition and/or interference.
[0096] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula I:
##STR00016##
[0097] wherein, independently for each occurrence,
[0098] n is 0 to 10 inclusive;
[0099] R.sup.1 is hydrogen or alkyl;
[0100] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl;
[0101] Y is --C(R.sub.b).sub.p--, --C(.dbd.O)-- or
--C(R.sub.b).sub.pC(.dbd.O)--;
[0102] X is --O--, --N(R.sub.a)-, --C(R.sub.b).sub.p-- or
--S--;
[0103] Z is alkyl, haloalkyl, --(CH.sub.2CH.sub.2O).sub.pR.sub.b or
--C(.dbd.O)R.sub.b;
[0104] p is 0 to 20 inclusive;
[0105] R.sub.a is hydrogen, alkyl, aryl or aralkyl;
[0106] R.sub.b is hydrogen, alkyl or haloalkyl; and denotes a
single bond, a cis double bond, or a trans double bond.
[0107] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula II:
##STR00017## [0108] wherein, independently for each occurrence,
[0109] n is 0 to 10 inclusive; [0110] R.sup.1 is hydrogen or alkyl;
[0111] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; [0112] Y is
--C(R.sub.b).sub.p--, --C(.dbd.O)-- or
--C(R.sub.b).sub.pC(.dbd.O)--; [0113] X is hydrogen, --O--, --S--,
--N(R.sub.a)-, --N(R.sub.a)--N(R.sub.a)-, --C(.dbd.O)--,
--C(.dbd.NR.sub.a)-, --C(.dbd.NOH)--, --C(.dbd.S)-- or
--C(R.sub.b).sub.p-; [0114] Z is absent, hydrogen, alkyl,
haloalkyl, aryl, aralkyl, --CN, --OR.sub.b,
--(CH.sub.2CH.sub.2O).sub.pR.sub.b, --C(.dbd.O)R.sub.b,
--C(.dbd.O)CH.sub.2F, --C(.dbd.O)CHF.sub.2, --C(.dbd.O)CF.sub.3,
--C(.dbd.O)CHN.sub.2, --C(.dbd.O)OR.sub.b, [0115]
--C(.dbd.O)CH.sub.2C(.dbd.O)R.sub.b,
--C(.dbd.O)C(.dbd.C(R.sub.b).sub.2)R.sub.b,
[0115] ##STR00018## [0116] p is 0 to 20 inclusive; [0117] R.sub.a
is hydrogen, alkyl, aryl or aralkyl; [0118] R.sub.b is hydrogen,
alkyl, haloalkyl, aryl or aralkyl; and denotes a single bond, a cis
double bond or a trans double bond.
[0119] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula III:
##STR00019## [0120] wherein, independently for each occurrence,
[0121] n is 0 to 10 inclusive; [0122] R.sup.1 is hydrogen or alkyl;
[0123] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; [0124] Y is
--CR.sub.b(OR.sub.b)-, --CR.sub.b(N(R.sub.a).sub.2)--,
--C(R.sub.b).sub.p--, --C(.dbd.O)-- or
--C(R.sub.b).sub.pC(.dbd.O)--; [0125] X is --O--, --S--,
--N(R.sub.a)-, --C(.dbd.O)--, or --C(R.sub.b).sub.p-; [0126] Z is
hydrogen, alkyl, haloalkyl, aryl, aralkyl, --OR.sub.b,
--N(R.sub.b).sub.2, --(CH.sub.2CH.sub.2O).sub.pR.sub.b,
--C(.dbd.O)R.sub.b, --C(.dbd.NR.sub.a)R.sub.b,
--C(.dbd.NOR.sub.b)R.sub.b, --C(OR.sub.b)(R.sub.b).sub.2,
--C(N(R.sub.a).sub.2)(R.sub.b).sub.2 or
--(CH.sub.2CH.sub.2O).sub.pR.sub.b; [0127] p is 0 to 20 inclusive;
[0128] R.sub.a is hydrogen, alkyl, aryl or aralkyl; [0129] R.sub.b
is hydrogen, alkyl, haloalkyl, aryl or aralkyl; and denotes a
single bond or a trans double bond.
[0130] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula VI:
##STR00020##
[0131] wherein, independently for each occurrence,
[0132] R.sup.1 is hydrogen, alkyl, aryl or aralkyl;
[0133] X is alkyl, alkenyl, --C(R.sub.b).sub.2-, --C(.dbd.O)--,
--C(.dbd.NR.sub.a)-, --C(OH)R.sub.b or
--C(N(R.sub.a).sub.2)R.sub.b--;
[0134] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl;
[0135] R.sub.a is hydrogen, alkyl, aryl or aralkyl; and
[0136] R.sub.b is hydrogen or alkyl.
[0137] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula I:
##STR00021##
[0138] wherein, independently for each occurrence,
[0139] n is 0 to 10 inclusive;
[0140] R.sup.1 is hydrogen or alkyl;
[0141] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl;
[0142] Y is --C(R.sub.b).sub.p--, --C(.dbd.O)-- or
--C(R.sub.b).sub.pC(.dbd.O)--;
[0143] X is --O--, --N(R.sub.a)-, --C(R.sub.b).sub.p-- or
--S--;
[0144] Z is alkyl, haloalkyl, --(CH.sub.2CH.sub.2O).sub.pR.sub.b or
--C(.dbd.O)R.sub.b;
[0145] p is 0 to 20 inclusive;
[0146] R.sub.a is hydrogen, alkyl, aryl or aralkyl;
[0147] R.sub.b is hydrogen, alkyl or haloalkyl; and denotes a
single bond, a cis double bond, or a trans double bond.
[0148] In certain embodiments, an inhibitor of isomerohydrolase
(IMH) has a structure represented by formula Ia, Ib, Ic, or Id:
##STR00022## [0149] wherein, independently for each occurrence,
[0150] n is 0 to 4 inclusive; [0151] R.sup.1 is hydrogen or alkyl;
[0152] R.sup.3 is hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, and sulfoxido; [0153] R.sup.4 is absent, hydrogen,
halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl,
aralkyenyl, aralkynyl, heteroaralkyl, heteroaralkyenyl,
heteroaralkynyl, cyano, nitro, sulfhydryl, hydroxyl, sulfonyl,
amino, acylamino, amido, alkylthio, carboxyl, carbamoyl, alkoxyl,
sulfonate, sulfate, sulfonamido, sulfamoyl, sulfonyl, and
sulfoxido; [0154] Y is --C(R.sub.b).sub.2- or --C(.dbd.O)--; [0155]
X is --O--, --N(R.sub.a)-, --C(R.sub.b).sub.2- or --S--; [0156] Z
is alkyl, haloalkyl or --C(.dbd.O)R.sub.b; [0157] R.sub.a is
hydrogen, alkyl, aryl or aralkyl; [0158] R.sub.b is hydrogen, alkyl
or haloalkyl; and denotes a single bond, a cis double bond, or a
trans double bond.
[0159] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein
R.sup.1 is methyl.
[0160] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein n is
0.
[0161] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein n is
1.
[0162] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein Y is
--CH.sub.2--.
[0163] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein X is
--O--.
[0164] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein X is
--N(H)--.
[0165] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein Z is
--C(.dbd.O)R.sub.b.
[0166] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein Z is
--C(.dbd.O)R.sub.b; and R.sub.b is haloalkyl.
[0167] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein Z is
alkyl.
[0168] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein Z is
haloalkyl.
[0169] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein
R.sup.3 is hydrogen.
[0170] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ia, Ib, Ic, or Id, wherein
R.sup.4 is hydrogen, methyl or absent.
[0171] In certain embodiments, an inhibitor of isomerohydrolase
(IMH) has a structure represented by formula Ie, If, Ig, or Ih:
##STR00023## [0172] wherein, independently for each occurrence,
[0173] n is 0 to 4 inclusive; [0174] R.sup.1 is hydrogen or alkyl;
[0175] R.sup.3 is hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, and sulfoxido; [0176] X is --O--, --N(R.sub.a)-,
--C(R.sub.b).sub.2- or --S--; [0177] Z is alkyl, haloalkyl or
--C(.dbd.O)R.sub.b; [0178] R.sub.a is hydrogen, alkyl, aryl or
aralkyl; and [0179] R.sub.b is hydrogen, alkyl or haloalkyl.
[0180] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein n is
0.
[0181] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein n is
1.
[0182] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein X is
--O--.
[0183] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein X is
--N(H)--.
[0184] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein Z is
--C(.dbd.O)R.sub.b.
[0185] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein Z is
--C(.dbd.O)R.sub.b; and R.sub.b is haloalkyl.
[0186] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein Z is
alkyl.
[0187] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein Z is
haloalkyl.
[0188] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein
R.sup.3 is hydrogen.
[0189] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein X is
--O--; and Z is alkyl.
[0190] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein X is
--O--; and Z is haloalkyl.
[0191] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein X is
--N(H)--; and Z is alkyl.
[0192] In further embodiments, an inhibitor of isomerohydrolase
(IMH) has the structure of formula Ie, If, Ig, or Ih, wherein X is
--N(H)--; and Z is haloalkyl.
[0193] In one embodiment, an inhibitor of isomerohydrolase (IMH) is
11-cis-retinyl bromoacetate (cBRA):
##STR00024##
[0194] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula II:
##STR00025## [0195] wherein, independently for each occurrence,
[0196] n is 0 to 10 inclusive; [0197] R.sup.1 is hydrogen or alkyl;
[0198] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; [0199] Y is
--C(R.sub.b).sub.p--, --C(.dbd.O)-- or
--C(R.sub.b).sub.pC(.dbd.O)--; [0200] X is hydrogen, --O--, --S--,
--N(R.sub.a)-, N(R.sub.a)--N(R.sub.a)-, --C(.dbd.O)--,
C(.dbd.NR.sub.a)-, --C(.dbd.NOH)--, --C(.dbd.S)-- or
--C(R.sub.b).sub.p-; [0201] Z is absent, hydrogen, alkyl,
haloalkyl, aryl, aralkyl, --CN, --OR.sub.b,
--(CH.sub.2CH.sub.2O).sub.pR.sub.b, --C(.dbd.O)R.sub.b,
--C(.dbd.O)CH.sub.2F, --C(.dbd.O)CHF.sub.2, --C(.dbd.O)CF.sub.3,
--C(.dbd.O)CHN.sub.2, --C(.dbd.O)OR.sub.b,
--C(.dbd.O)CH.sub.2C(.dbd.O)R.sub.b,
--C(.dbd.O)C(.dbd.C(R.sub.b).sub.2)R.sub.b,
[0201] ##STR00026## [0202] p is 0 to 20 inclusive; [0203] R.sub.a
is hydrogen, alkyl, aryl or aralkyl; [0204] R.sub.b is hydrogen,
alkyl, haloalkyl, aryl or aralkyl; and denotes a single bond, a cis
double bond or a trans double bond.
[0205] In certain embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIe, or IId:
##STR00027## [0206] wherein, independently for each occurrence,
[0207] n is 0 to 4 inclusive; [0208] R.sup.1 is hydrogen or alkyl;
[0209] R.sup.3 is hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, and sulfoxido; [0210] R.sup.4 is absent, hydrogen,
halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl,
aralkyenyl, aralkynyl, heteroaralkyl, heteroaralkyenyl,
heteroaralkynyl, cyano, nitro, sulfhydryl, hydroxyl, sulfonyl,
amino, acylamino, amido, alkylthio, carboxyl, carbamoyl, alkoxyl,
sulfonate, sulfate, sulfonamido, sulfamoyl, sulfonyl, and
sulfoxido; [0211] Y is --C(.dbd.O)-- or --C(R.sub.b).sub.2-; [0212]
X is hydrogen, --O--, --S--, --N(R.sub.a)-,
--N(R.sub.a)--N(R.sub.a)-, --C(.dbd.O)--, --C(.dbd.NR.sub.a)-,
--C(.dbd.NOH)--, --C(.dbd.S)-- or --C(R.sub.b).sub.2-; [0213] Z is
absent, hydrogen, alkyl, haloalkyl, aryl, aralkyl, --CN,
--OR.sub.b, --C(.dbd.O)R.sub.b, --C(.dbd.O)CH.sub.2F,
--C(.dbd.O)CHF.sub.2, --C(.dbd.O)CF.sub.3, --C(.dbd.O)CHN.sub.2,
--C(.dbd.O)CH.sub.2OC(.dbd.O)R.sub.b, --C(.dbd.O)OR.sub.b,
--C(.dbd.O)C(.dbd.C(R.sub.b).sub.2)R.sub.b,
[0213] ##STR00028## [0214] R.sub.a is hydrogen, alkyl, aryl or
aralkyl; [0215] R.sub.b is hydrogen, alkyl, haloalkyl, aryl or
aralkyl; and denotes a single bond, a cis double bond or a trans
double bond.
[0216] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein n is 0.
[0217] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein n is 1.
[0218] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein R.sup.1 is hydrogen or methyl.
[0219] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein R.sup.3 is hydrogen.
[0220] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein R.sup.4 is hydrogen or methyl.
[0221] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein Y is --CH.sub.2--
[0222] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein X is --O--.
[0223] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein X is --NH--.
[0224] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein X is --C(R.sub.b).sub.2-.
[0225] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein X is --C(.dbd.O)--.
[0226] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein Z is alkyl.
[0227] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIa,
IIb, IIc, or IId, wherein Z is haloalkyl.
[0228] In certain embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh:
##STR00029## [0229] wherein, independently for each occurrence,
[0230] n is 0 to 4 inclusive; [0231] R.sup.1 is hydrogen or alkyl;
[0232] R.sup.3 is hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl,
heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, and sulfoxido; [0233] X is hydrogen, --O--, --S--,
--N(R.sub.a)-, --N(R.sub.a)--N(R.sub.a)-, --C(.dbd.O)--,
--C(.dbd.NR.sub.a)-, --C(.dbd.NOH)--, --C(.dbd.S)-- or
--C(R.sub.b).sub.2-; [0234] Z is absent, hydrogen, alkyl,
haloalkyl, aryl, aralkyl, --CN, --OR.sub.b, --C(.dbd.O)R.sub.b,
--C(.dbd.O)CH.sub.2F, --C(.dbd.O)CHF.sub.2, --C(.dbd.O)CF.sub.3,
--C(.dbd.O)CHN.sub.2, --C(.dbd.O)CH.sub.2C(.dbd.O)R.sub.b,
--C(.dbd.O)OR.sub.b,
--C(.dbd.O)C(.dbd.C(R.sub.b).sub.2)R.sub.b,
[0234] ##STR00030## [0235] R.sub.a is hydrogen, alkyl, aryl or
aralkyl; and [0236] R.sub.b is hydrogen, alkyl, haloalkyl, aryl or
aralkyl.
[0237] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein n is 0.
[0238] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein n is 1.
[0239] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein R.sup.1 is hydrogen or methyl.
[0240] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein R.sup.3 is hydrogen.
[0241] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein R.sup.4 is hydrogen or methyl.
[0242] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein X is --O--.
[0243] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein X is --NH--.
[0244] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein X is --CH.sub.2--.
[0245] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein X is --C(.dbd.O)--.
[0246] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein Z is alkyl.
[0247] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein Z is haloalkyl.
[0248] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein Z is --C(.dbd.O)R.sub.b.
[0249] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein X is --O--; and Z is
--C(.dbd.O)R.sub.b--
[0250] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula IIe,
IIf, IIg, or IIh, wherein X is --CH.sub.2--; and Z is
--C(.dbd.O)R.sub.b.
[0251] In further embodiments, an inhibitor of lecithin retinol
acyl transferase (LRAT) has a structure represented by formula Ie,
IIf, IIg, or IIh, wherein X is --NH--; and Z is
--C(.dbd.O)R.sub.b.
[0252] In one embodiment, an inhibitor of lecithin retinol acyl
transferase (LRAT) is 13-desmethyl-13,14-dihydro-all-trans-retunyl
trifluoroacetate (RFA):
##STR00031##
[0253] In one embodiment, an inhibitor of lecithin retinol acyl
transferase (LRAT) is all-trans-retinyl .alpha.-bromoacetate.
[0254] In certain embodiments, an inhibitor of isomerohydrolase
(IMH), an inhibitor 11-cis-retinol dehydrogenase, an inhibitor of
lecithin retinol acyl transferase (LRAT), or an antagonist of
chaperone retinal pigment epithelium (RPE65) has a structure
represented by formula III:
##STR00032## [0255] wherein, independently for each occurrence,
[0256] n is 0 to 10 inclusive; [0257] R.sup.1 is hydrogen or alkyl;
[0258] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; [0259] Y is
--CR.sub.b(OR.sub.b)-, CR.sub.b(N(R.sub.a).sub.2)--,
--C(R.sub.b).sub.p--, --C(.dbd.O)-- or
--C(R.sub.b).sub.pC(.dbd.O)--; [0260] X is --O--, --S--,
--N(R.sub.a)-, --C(.dbd.O)--, or --C(R.sub.b).sub.p-; [0261] Z is
hydrogen, alkyl, haloalkyl, aryl, aralkyl, --OR.sub.b,
--N(R.sub.b).sub.2, --(CH.sub.2CH.sub.2O).sub.pR.sub.b,
--C(.dbd.O)R.sub.b, --C(.dbd.NR.sub.a)R.sub.b,
--C(.dbd.NOR.sub.b)R.sub.b, --C(OR.sub.b)(R.sub.b).sub.2,
--C(N(R.sub.a).sub.2)(R.sub.b).sub.2 or
--(CH.sub.2CH.sub.2O).sub.pR.sub.b; [0262] p is 0 to 20 inclusive;
[0263] R.sub.a is hydrogen, alkyl, aryl or aralkyl; [0264] R.sub.b
is hydrogen, alkyl, haloalkyl, aryl or aralkyl; and denotes a
single bond or a trans double bond.
[0265] In certain embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc or IIId:
##STR00033## [0266] wherein, independently for each occurrence,
[0267] n is 0 to 4 inclusive; [0268] R.sup.1 is hydrogen or alkyl;
[0269] Y is --C(.dbd.O)--, --CR.sub.b(OR.sub.b)-,
--CR.sub.b(N(R.sub.a).sub.2)- or --C(R.sub.b).sub.2-; [0270] X is
--O--, --S--, --N(R.sub.a)-, --C(.dbd.O)--, or --C(R.sub.b).sub.2-;
[0271] Z is hydrogen, alkyl, haloalkyl, aryl, aralkyl, --OR.sub.b,
--N(R.sub.b).sub.2, --C(.dbd.O)R.sub.b, C(.dbd.NR.sub.a)R.sub.b,
--C(.dbd.NOH)R.sub.b, --C(OR.sub.b)(R.sub.b).sub.2,
C(N(R.sub.a).sub.2)(R.sub.b).sub.2 or
--(CH.sub.2CH.sub.2O).sub.pR.sub.b; [0272] R.sub.a is hydrogen,
alkyl, aryl or aralkyl; [0273] R.sub.b is hydrogen, alkyl,
haloalkyl, aryl or aralkyl; [0274] p is 0 to 10 inclusive; and
denotes a single bond or a trans double bond.
[0275] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein n is 0.
[0276] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein n is 1.
[0277] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein R.sup.1 is hydrogen or methyl.
[0278] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein R.sup.3 is hydrogen.
[0279] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein R.sup.4 is hydrogen or methyl.
[0280] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein X is --O--.
[0281] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein X is --NH--.
[0282] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIc, or IIId, wherein X is --C(R.sub.b).sub.2-.
[0283] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIe, or IIId, wherein X is --C(.dbd.O)--.
[0284] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein Z is alkyl.
[0285] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIa,
IIIb, IIIc, or IIId, wherein Z is haloalkyl.
[0286] In certain embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh:
##STR00034## [0287] wherein, independently for each occurrence,
[0288] n is 0 to 4 inclusive; [0289] R.sup.1 is hydrogen or alkyl;
[0290] X is --O--, --S--, --N(R.sub.a)-, --C(.dbd.O)--, or
--C(R.sub.b).sub.2-; [0291] Z is hydrogen, alkyl, haloalkyl, aryl,
aralkyl, --OR.sub.b, --N(R.sub.b).sub.2, --C(.dbd.O)R.sub.b,
--C(.dbd.NR.sub.a)R.sub.b, --C(.dbd.NOH)R.sub.b,
--C(OR.sub.b)(R.sub.b).sub.2, --C(N(R.sub.a).sub.2)(R.sub.b).sub.2
or --(CH.sub.2CH.sub.2O).sub.pR.sub.b; [0292] R.sub.a is hydrogen,
alkyl, aryl or aralkyl; [0293] R.sub.b is hydrogen, alkyl,
haloalkyl, aryl or aralkyl; and [0294] p is 0 to 10 inclusive.
[0295] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein n is 0.
[0296] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein n is 1.
[0297] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein R.sup.1 is hydrogen or methyl.
[0298] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Y is --C(.dbd.O)--.
[0299] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Y is --CH.sub.2--.
[0300] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Z is --C(.dbd.O)R.sub.b.
[0301] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Z is --CH(OH)R.sub.b--.
[0302] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Z is CH(NH)R.sub.b.
[0303] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Z is alkyl.
[0304] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IIIe,
IIIf, IIIg, or IIIh, wherein Z is haloalkyl.
[0305] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is 13-cis-retinoic acid (isoretinoin,
ACCUTANE.RTM.):
##STR00035##
[0306] In certain embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV:
##STR00036##
[0307] wherein, independently for each occurrence,
[0308] n is 1, 2, 3 or 4;
[0309] Y is --C(R.sub.b).sub.2-, --C(.dbd.O)-- or
--OC(.dbd.O)--;
[0310] X is --O--, --NR.sub.a--, --C(R.sub.b).sub.2- or
--C(.dbd.O)--;
[0311] Z is --C(.dbd.O)R.sub.b, --OR.sub.b, --N(R.sub.b).sub.2,
alkyl or haloalkyl;
[0312] R.sub.a is hydrogen, alkyl, haloalkyl, aryl or aralkyl;
and
[0313] R.sub.b is hydrogen, alkyl, haloalkyl, aryl or aralkyl.
[0314] In further embodiments, an inhibitor of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Y is --CH.sub.2--.
[0315] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein X is --O--.
[0316] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Z is --C(.dbd.O)R.sub.b; and R.sub.b is alkyl.
[0317] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Z is alkyl.
[0318] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Y is --CH.sub.2--; X is --O--; Z is --C(.dbd.O)R.sub.b; and
R.sub.b is alkyl.
[0319] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Y is --CH.sub.2--; X is --O--; and Z is alkyl.
[0320] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Y is --CH.sub.2--; X is --C(.dbd.O)--; and Z is alkyl.
[0321] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula IV,
wherein Y is --CH.sub.2--; X is --C(.dbd.O)--; Z is
--N(R.sub.b).sub.2; and R.sub.b is alkyl.
[0322] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is geranyl palmitate (K.sub.D=301 nM):
##STR00037##
[0323] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is farnesyl palmitate (K.sub.D=63 nM)
##STR00038##
[0324] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is geranylgeranyl palmitate (K.sub.D=213
nM):
##STR00039##
[0325] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is geranyl palmityl ether (K.sub.D=416 nM):
##STR00040##
[0326] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is farnesyl palmityl ether (K.sub.D=60 nM):
##STR00041##
[0327] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is geranylgeranyl palmityl ether (K.sub.D=195
nM):
##STR00042##
[0328] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound:
##STR00043##
[0329] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=96 nM):
##STR00044##
[0330] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound:
##STR00045##
[0331] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=56 nM):
##STR00046##
[0332] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is farnesyl octyl ketone:
##STR00047##
[0333] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is octyl farnesimide:
##STR00048##
[0334] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is palmityl farnesimide:
##STR00049##
[0335] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=56 nM):
##STR00050##
[0336] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=58.+-.5 nM),
called 13,17,21-Trimethyl-docosa-12,16,20-trien-11-one or
"TDT":
##STR00051##
[0337] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=96.+-.14 nM),
called 3,7,11-Trimethyl-dodeca-2,6,10-trienoic acid hexadecylamide
or "TDH":
##STR00052##
[0338] In certain embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V:
##STR00053##
[0339] wherein, independently for each occurrence,
[0340] n is 1, 2 or 3;
[0341] Y is --C(R.sub.b).sub.2-, --C(.dbd.O)-- or --CH(OH)--;
[0342] X is --O--, NR.sub.a, or --C(R.sub.b).sub.2-;
[0343] Z is --C(.dbd.O)R.sub.b, hydrogen,
--(CH.sub.2CH.sub.2O).sub.pR.sub.b, alkyl or haloalkyl;
[0344] R.sub.a is hydrogen, alkyl, haloalkyl, aryl or aralkyl;
[0345] R.sub.b is hydrogen, alkyl, haloalkyl, aryl or aralkyl;
and
[0346] p is 1 to 10 inclusive.
[0347] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein Y is --CH.sub.2--.
[0348] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein Y is --C(.dbd.O)--.
[0349] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein Y is --CH(OH)--.
[0350] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein X is --O--.
[0351] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein X is --NR.sub.a--
[0352] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein X is --C(R.sub.b)-.
[0353] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein Z is alkyl.
[0354] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein Z is --C(.dbd.O)R.sub.b; and R.sub.b is alkyl.
[0355] In further embodiments, an antagonist of retinal pigment
epithelium (RPE65) has a structure represented by formula V,
wherein Z is --(CH.sub.2CH.sub.2O).sub.pR.sub.b; and R.sub.b is
alkyl.
[0356] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is .beta.-ionoacetyl palmitate (K.sub.D=153
nM):
##STR00054##
[0357] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is .beta.-ionoacetyl palmityl ether (K.sub.D=156
nM):
##STR00055##
[0358] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is retinyl palmitate (4a; K.sub.D=47 nM):
##STR00056##
[0359] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is retinyl hexanoate (4b; K.sub.D=235 nM):
##STR00057##
[0360] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is retinyl pentanoate:
##STR00058##
[0361] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is retinyl acetate (4c; K.sub.D=1,300 nM):
##STR00059##
[0362] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is palmityl retinyl ether (4d, K.sub.D=25
nM):
##STR00060##
[0363] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is hexyl retinyl ether (K.sub.D=151 nM):
##STR00061##
[0364] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is methyl retinyl ether (K.sub.D=24 nM):
##STR00062##
[0365] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is retinyl [2-(2'-methoxy)ethoxy]ethyl ether
(K.sub.D=486 nM):
##STR00063##
[0366] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is:
##STR00064##
[0367] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is N-palmityl retinimide (K.sub.D=40 nM):
##STR00065##
[0368] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is N,N-dimethyl retinimide (K.sub.D=3,577
nM):
##STR00066##
[0369] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is N-tert-butyl retinimide (K.sub.D=4,321
nM):
##STR00067##
[0370] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is palmityl retinyl alcohol (K.sub.D=170
nM):
##STR00068##
[0371] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is methyl retinyl alcohol:
##STR00069##
[0372] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is palmityl retinyl ketone (K.sub.D=64 nM):
##STR00070##
[0373] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is retinyl decyl ketone:
##STR00071##
[0374] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is methyl retinyl ketone (K.sub.D=3,786 nM):
##STR00072##
[0375] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (4e):
##STR00073##
[0376] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (4f; K.sub.D=64
nM)
##STR00074##
[0377] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=173 nM):
##STR00075##
[0378] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is the following compound (K.sub.D=3,786
nM):
##STR00076##
[0379] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is:
##STR00077##
[0380] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is:
##STR00078##
[0381] In one embodiment, an antagonist of retinal pigment
epithelium (RPE65) is:
##STR00079##
[0382] The above-described RPE65 antagonist compounds and general
formulas of compounds, with their various substituent definitions
and further embodiments, are also LRAT inhibitors, and are
incorporated herein by reference as LRAT inhibitors.
[0383] Other antagonists of RPE65 and inhibitors of LRAT include
agents that inhibit palmitoylation. For example, 2-bromopalmitate
inhibits palmitoylation. In some embodiments, a racemic mixture of
2-bromopalmitate may be applied to inhibit LRAT and/or antagonize
RPE65. In other embodiments, purified (R)-2-bromopalmitic acid may
be applied to inhibit LRAT and/or antagonize RPE65. In yet other
embodiments, purified (S)-2-bromopalmitic acid may be applied to
inhibit LRAT and/or antagonize RPE65.
[0384] In certain embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VI:
##STR00080##
[0385] wherein, independently for each occurrence,
[0386] R.sup.1 is hydrogen, alkyl, aryl or aralkyl;
[0387] X is alkyl, alkenyl, --C(R.sub.b).sub.2-, --C(.dbd.O)--,
--C(.dbd.NR.sub.a)-, --C(OH)R.sub.b or
--C(N(R.sub.a).sub.2)R.sub.b--;
[0388] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl;
[0389] R.sub.a is hydrogen, alkyl, aryl or aralkyl; and
[0390] R.sub.b is hydrogen or alkyl.
[0391] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VI, wherein
R.sup.1 is hydrogen.
[0392] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VI, wherein X
is --C(R.sub.b).sub.2-.
[0393] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VI, wherein X
is --C(.dbd.O)--.
[0394] In certain embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIa or
VIb:
##STR00081##
[0395] wherein, independently for each occurrence,
[0396] R.sup.1 is hydrogen, alkyl, aryl or aralkyl;
[0397] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl;
[0398] R.sup.3 is hydrogen or alkyl;
[0399] R.sub.a is hydrogen, alkyl, aryl or aralkyl;
[0400] R.sub.b is hydrogen or alkyl; and denotes a single bond, a
cis double bond, or a trans double bond.
[0401] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIa or VIb,
wherein R.sup.1 is hydrogen.
[0402] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIa or VIb,
wherein R.sup.2 is alkyl.
[0403] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIa or VIb,
wherein R.sup.3 is hydrogen or methyl.
[0404] In certain embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIc, VId or
VIe:
##STR00082## [0405] wherein, independently for each occurrence,
[0406] n is 1 to 5 inclusive; [0407] m is 0 to 30 inclusive; [0408]
R.sup.1 is hydrogen, alkyl, aryl or aralkyl; [0409] R.sup.2 is
hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,
or aralkyl; [0410] R.sup.3 is hydrogen or alkyl; [0411] R.sup.4 is
hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl,
aralkyl, aralkyenyl, aralkynyl, heteroaralkyl, heteroaralkyenyl,
heteroaralkynyl, cyano, nitro, sulfhydryl, hydroxyl, sulfonyl,
amino, acylamino, amido, alkylthio, carboxyl, carbamoyl, alkoxyl,
sulfonate, sulfate, sulfonamido, sulfamoyl, sulfonyl, and
sulfoxido; [0412] R.sub.a is hydrogen, alkyl, aryl or aralkyl;
and
[0413] R.sub.b is hydrogen or alkyl.
[0414] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIc, wherein
R.sup.1 is hydrogen.
[0415] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIc, wherein
R.sup.4 is hydrogen.
[0416] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIe, wherein
R.sup.1 is hydrogen; and R.sup.4 is hydrogen.
[0417] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VId, wherein n
is 1, 2 or 3.
[0418] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VId, wherein
R.sup.3 is methyl.
[0419] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VId, wherein
R.sup.1 is hydrogen.
[0420] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VId, wherein n
is 1, 2 or 3; R.sup.3 is methyl.
[0421] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VId, wherein n
is 1, 2 or 3; R.sup.3 is methyl; and R.sup.1 is hydrogen.
[0422] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIe, wherein
R.sup.1 is hydrogen.
[0423] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIe, wherein m
is 1 to 10 inclusive.
[0424] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIe, wherein m
is 11 to 20 inclusive.
[0425] In further embodiments, an inhibitor of 11-cis-retinol
dehydrogenase has a structure represented by formula VIe, wherein m
is 11 to 20 inclusive; and R.sup.1 is hydrogen.
[0426] 11-cis-retinol dehydrogenase inhibitors having structures
represented by formula VIe may be generated according to a
diversity library approach as shown in Scheme 1, among other
ways:
##STR00083##
[0427] In one embodiment, an inhibitor of 11-cis-retinol
dehydrogenas is 13-cis-retinoic acid (isoretinoin,
ACCUTANE.RTM.):
##STR00084##
[0428] Also included are pharmaceutically acceptable addition salts
and complexes of the compounds of the formulas given above. In
cases wherein the compounds may have one or more chiral centers,
unless specified, the compounds contemplated herein may be a single
stereoisomer or racemic mixtures of stereoisomers. Further included
are prodrugs, analogs, and derivatives thereof.
[0429] In some embodiments, two or more enzyme inhibitors and/or
RPE65 binding inhibitors may be combined. In some embodiments, an
enzyme inhibitor and/or RPE65 binding inhibitor may be combined
with a short-circuiting compound. Combinations may be selected to
inhibit sequential steps in the visual cycle (that is, two steps
that occur one immediately after the other).
[0430] In certain embodiments, an inhibitor of isomerohydrolase
(IMH) may be a compound having a structure represented by general
structure 1:
##STR00085## [0431] wherein, independently for each occurrence:
[0432] R, R.sub.1, R.sub.2, and R.sub.3 are H, alkyl, alkenyl,
alkynyl, aryl, aralkyl, heteroaryl, or heteroaralkyl; [0433] W and
Y are O, NR, R, or S; [0434] X is H, alkyl, haloalkyl, aryl, or
halide; [0435] m and n are integers from 1 to 6 inclusive; and
[0436] p is an integer from 0 to 6 inclusive.
[0437] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me.
[0438] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein m is
2.
[0439] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein n is
2.
[0440] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein W is
O.
[0441] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein W is
C.
[0442] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein Y is
O.
[0443] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein p is
1.
[0444] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein X is
Br.
[0445] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, and m is 2.
[0446] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 2, and n is 2.
[0447] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 2, n is 2, and W is O.
[0448] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 2, n is 2, W is O, and Y is
O.
[0449] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 2, n is 2, W is O, Y is O, and
p is 1.
[0450] In a further embodiment, the inhibitor of IMH has the
structure of formula 1 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 2, n is 2, W is O, Y is O, p
is 1, and X is Br.
[0451] In one embodiment, an isomerohydrolase inhibitor is
11-cis-retinyl bromoacetate (cRBA):
##STR00086##
[0452] In certain embodiments, an inhibitor of IMH may be a
compound of formula 8a:
##STR00087## [0453] wherein, independently for each occurrence:
[0454] X is O, S, NR', CH.sub.2, or NHNR'; [0455] Z is O or NOH;
[0456] R.sub.1 is --CH.sub.2F, --CHF.sub.2, --CF.sub.3,
--CH.sub.2N.sub.2, --CH.sub.2C(O)OR, --OR', --C(O)CHR',
--C(NH)CHR', or --CH.dbd.CHR'; [0457] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0457] ##STR00088## [0458] R''' is CH.sub.3 or H; and [0459] n is
0, 1 or 2; [0460] wherein denotes a single bond, a cis double bond
or a trans double bond.
[0461] Compounds of formula 8a may be considered irreversible
inhibitors of IMH because they can covalently bind IMH, permanently
disabling it.
[0462] In certain embodiments, an inhibitor of IMH may be a
compound of formula 8a wherein Z is O.
[0463] In certain embodiments, an inhibitor of IMH may be a
compound of formula 8b:
##STR00089## [0464] wherein, independently for each occurrence:
[0465] Y is C.dbd.O, C.dbd.S, C.dbd.NR', or CH.sub.2; [0466]
R.sub.1 is R', --OR', or --CN; [0467] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0467] ##STR00090## [0468] R''' is CH.sub.3 or H; and [0469] n is
0, 1, or 2; [0470] wherein denotes a single bond, a cis double bond
or a trans double bond.
[0471] Compounds of formula 8b may be considered reversible
inhibitors of IMH because they can noncovalently bind IMH without
permanently disabling it.
[0472] In certain embodiments, an inhibitor of IMH may be a
compound of formula 8c:
##STR00091## [0473] wherein, independently for each occurrence:
[0474] X is O, S, NR', CH.sub.2, or NHNR'; [0475] Z is O or NOH;
[0476] R.sub.1 is --CH.sub.2F, --CHF.sub.2, --CF.sub.3,
--CH.sub.2N.sub.2, --CH.sub.2C(O)OR', --OR', --C(O)CHR',
--C(NH)CHR', or --CH.dbd.CHR'; [0477] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0477] ##STR00092## [0478] R''' is CH.sub.3 or H; and [0479] n is
0, 1 or 2.
[0480] Compounds of formula 8c may be considered irreversible
inhibitors of IMH because they can covalently bind IMH, permanently
disabling it.
[0481] In certain embodiments, an inhibitor of IMH may be a
compound of formula 8c wherein Z is O.
[0482] In certain embodiments, an inhibitor of IMH may be a
compound of formula 8d:
##STR00093## [0483] wherein, independently for each occurrence:
[0484] Y is C.dbd.O, C.dbd.S, C.dbd.NR', or CH.sub.2; [0485]
R.sub.1 is R', --OR', or --CN; [0486] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0486] ##STR00094## [0487] R''' is CH.sub.3 or H; and [0488] n is
0, 1 or 2.
[0489] Compounds of formula 8d may be considered reversible
inhibitors of IMH because they can noncovalently bind IMH without
permanently disabling it.
[0490] In certain embodiments, an inhibitor of LRAT may be a
compound having a structure represented by general structure 2:
##STR00095## [0491] wherein, independently for each occurrence:
[0492] R, R.sub.1, R.sub.2, and R.sub.3 are H, alkyl, alkenyl,
alkynyl, aryl, aralkyl, heteroaryl, or heteroaralkyl; [0493] W and
Y are O, NR, R, or S; [0494] X is H, alkyl, haloalkyl, or aryl;
[0495] m and n are integers from 1 to 6 inclusive; and [0496] p is
an integer from 0 to 6 inclusive.
[0497] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me.
[0498] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein m is
3.
[0499] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein n is
1.
[0500] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein W is
O.
[0501] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein W is
C.
[0502] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein Y is
O.
[0503] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein p is
0.
[0504] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein X is
OCF.sub.3.
[0505] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, and m is 3.
[0506] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 3, and n is 1.
[0507] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 3, n is 1, and W is O.
[0508] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 3, n is 1, W is O, and Y is
O.
[0509] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 3, n is 1, W is O, Y is O, and
p is 0.
[0510] In a further embodiment, the inhibitor of LRAT has the
structure of formula 2 and the attendant definitions, wherein
R.sub.2 and R.sub.3 is H or Me, m is 3, n is 1, W is O, Y is O, p
is 0, and X is OCF.sub.3.
[0511] An exemplary inhibitor of LRAT is all-trans-retinyl
.alpha.-bromoacetate. Another exemplary inhibitor of LRAT is
13-desmethyl-13,14-dihydro-all-trans-retinyl trifluoroacetate
(RFA):
##STR00096##
[0512] In certain embodiments, a compound that interferes with
RPE65 binding may be a compound having a structure represented by
general structure 3:
##STR00097## [0513] wherein, independently for each occurrence:
[0514] R and R.sub.1 are H, alkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, or heteroaralkyl; [0515] R.sub.2 is H, alkyl, alkenyl,
alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, or --CO.sub.2R;
[0516] R.sub.3 is H, alkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, heteroaralkyl, or --CH.sub.2OR.sub.4; [0517] R.sub.4 is
H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, heterocyclyl; and [0518] m is an integer from 1 to 6
inclusive.
[0519] In a further embodiment, the inhibitor of LRAT has the
structure of formula 3 and the attendant definitions, wherein
R.sub.2 is H, Me, or --CO.sub.2H.
[0520] In a further embodiment, the inhibitor of LRAT has the
structure of formula 3 and the attendant definitions, wherein m is
4.
[0521] In a further embodiment, the inhibitor of LRAT has the
structure of formula 3 and the attendant definitions, wherein
R.sub.3 is H.
[0522] In a further embodiment, the inhibitor of LRAT has the
structure of formula 3 and the attendant definitions, wherein
R.sub.2 is H, Me, or --CO.sub.2H and m is 4.
[0523] In a further embodiment, the inhibitor of LRAT has the
structure of formula 3 and the attendant definitions, wherein
R.sub.2 is H, Me, or --CO.sub.2H, m is 4, and R.sub.3 is H.
[0524] In certain embodiments, an inhibitor of LRAT may be a
compound of formula 6a:
##STR00098## [0525] wherein, independently for each occurrence:
[0526] X is O, S, NR', CH.sub.2, or NHNR'; [0527] Z is O or
NOH;
[0528] R.sub.1 is --CH.sub.2F, --CHF.sub.2, --CF.sub.3,
--CH.sub.2N.sub.2, --CH.sub.2C(O)OR, --OR', --C(O)CHR',
--C(NH)CHR', or --CH.dbd.CHR'; [0529] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0529] ##STR00099## [0530] n is 1, 2, or 3; [0531] wherein denotes
a single bond, a cis double bond or a trans double bond.
[0532] Compounds of formula 6a may be considered irreversible
inhibitors of LRAT because they can covalently bind LRAT,
permanently disabling it.
[0533] In certain embodiments, an inhibitor of LRAT may be a
compound of formula 6a wherein Z is O.
[0534] In certain embodiments, an inhibitor of LRAT may be a
compound of formula 6b:
##STR00100## [0535] wherein, independently for each occurrence:
[0536] Y is C.dbd.O, C.dbd.S, C.dbd.NR', or CH.sub.2; [0537]
R.sub.1 is R', --OR', or --CN; [0538] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0538] ##STR00101## [0539] n is 1, 2, or 3; [0540] wherein denotes
a single bond, a cis double bond or a trans double bond.
[0541] Compounds of formula 6c may be considered reversible
inhibitors of LRAT because they can noncovalently bind LRAT without
permanently disabling it.
[0542] In certain embodiments, an inhibitor of LRAT may be a
compound of formula 6c:
##STR00102## [0543] wherein independently for each occurrence:
[0544] X is O, S, NR', CH.sub.2, or NHNR'; [0545] Z is O or
NOH;
[0546] R.sub.1 is --CH.sub.2F, --CHF.sub.2, --CF.sub.3,
--CH.sub.2N.sub.2, --CH.sub.2C(O)OR', --OR', --C(O)CHR',
--C(NH)CHR', or --CH.dbd.CHR'; [0547] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0547] ##STR00103## [0548] n is 1, 2, or 3.
[0549] Compounds of formula 6c may be considered irreversible
inhibitors of LRAT because they can covalently bind LRAT,
permanently disabling it.
[0550] In certain embodiments, an inhibitor of LRAT may be a
compound of formula 6c wherein Z is O.
[0551] In certain embodiments, an inhibitor of LRAT may be a
compound of formula 6d:
##STR00104##
[0552] wherein, independently for each occurrence:
[0553] Y is C.dbd.O, C.dbd.S, C.dbd.NR', or CH.sub.2;
[0554] R.sub.1 is R', --OR', or --CN;
[0555] R' is H, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or
heteroaralkyl;
##STR00105##
[0556] n is 1, 2, or 3.
[0557] Compounds of formula 6d may be considered reversible
inhibitors of LRAT because they can noncovalently bind LRAT without
permanently disabling it.
[0558] One exemplary embodiment of a compound that interferes with
RPE65 binding is 13-cis-retinoic acid (isotretinoin,
ACCUTANE.RTM.):
##STR00106##
[0559] 13-cis-retinoic acid is converted in vivo to
all-trans-retinoic acid, which is a powerful inhibitor of RPE65
function.
[0560] In certain embodiments, an antagonist of RPE65 is a compound
having a structure represented by general structure 4:
##STR00107## [0561] wherein, independently for each occurrence:
[0562] R, R.sub.1, R.sub.2 are H, alkyl, alkenyl, alkynyl, aryl,
aralkyl, heteroaryl, heteroaralkyl, alkoxy, aryloxy, amino, halo,
hydroxy, or carboxyl; [0563] R.sub.3 is alkyl, alkenyl, alkynyl,
aryl, aralkyl, heteroaryl, heteroaralkyl, or ether;
[0564] L is H, OH, NH.sub.2, N(R).sub.2, alkoxy, aryloxy, halo,
hydroxy, carboxyl, or two L taken together represent O, S, or NR;
[0565] X is C(R).sub.2, O, S, or NR; and [0566] m is an integer
from 1 to 6 inclusive.
[0567] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O.
[0568] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
CH.sub.2.
[0569] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
NH.
[0570] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein two
Ls taken together represent O. In a further embodiment, an RPE65
antagonist has the structure of formula 4 and the attendant
definitions, wherein two Ls taken together represent NOH.
[0571] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein L is
H, OH, or NH.sub.2.
[0572] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein each
L is H.
[0573] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein m is
4.
[0574] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein m is
3.
[0575] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein
R.sub.2 is H or methyl.
[0576] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein
R.sub.3 is alkyl.
[0577] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein
R.sub.3 is ether.
[0578] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O and two L taken together represents O.
[0579] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O and each L is H.
[0580] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
NH and two L taken together represents O.
[0581] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
CH.sub.2 and two L taken together represents O.
[0582] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
CH.sub.2 and two L taken together represents NOH.
[0583] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O, two L taken together represent O, R.sub.2 is H or methyl, m is
4, and R.sub.3 is a C15 alkyl.
[0584] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O, two L taken together represent O, R.sub.2 is H or methyl, m is
4, and R.sub.3 is a C5 alkyl.
[0585] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O, two L taken together represent O, R.sub.2 is H or methyl, m is
4, and R.sub.3 is methyl.
[0586] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O, each L is H, R.sub.2 is H or methyl, m is 4, and R.sub.3 is a
C15 alkyl.
[0587] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
NH, two L taken together represents O, R.sub.2 is H or methyl, m is
4, and R.sub.3 is a C15 alkyl.
[0588] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
CH.sub.2, two L taken together represents O, R.sub.2 is H or
methyl, m is 4, and R.sub.3 is a C15 alkyl.
[0589] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O, each L is H, R.sub.2 is H or methyl, m is 4, and R.sub.3 is an
ether.
[0590] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
O, each L is H, R.sub.2 is H or methyl, m is 4, and R.sub.3 is
--CH.sub.2OCH.sub.2CH.sub.2OCH.sub.2CH.sub.2OC.sub.7H.sub.15.
[0591] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
CH.sub.2, two L taken together represent NOH, R.sub.2 is H or
methyl, m is 4, and R.sub.3 is a C15 alkyl.
[0592] In a further embodiment, an RPE65 antagonist has the
structure of formula 4 and the attendant definitions, wherein X is
CH.sub.2, L is H, OH, or NH.sub.2, R.sub.2 is H or methyl, m is 4,
and R.sub.3 is a C15 alkyl.
[0593] In certain embodiments, an inhibitor of RPE65 may be a
compound of formula 7a:
##STR00108## [0594] wherein, independently for each occurrence:
[0595] X is O, S, NR', CH.sub.2, or NHNR'; [0596] Z is O or
NOH;
[0597] R.sub.1 is --CH.sub.2F, --CHF.sub.2, --CF.sub.3,
--CH.sub.2N.sub.2, --CH.sub.2C(O)OR', --OR', --C(O)CHR',
--C(NH)CHR', or --CH.dbd.CHR'; [0598] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0598] ##STR00109## [0599] n is 1, 2, or 3; [0600] wherein denotes
a single bond, a cis double bond or a trans double bond.
[0601] Compounds of formula 7a may be considered irreversible
antagonists of RPE65 because they can covalently bind RPE65,
permanently disabling it.
[0602] In certain embodiments, an inhibitor of RPE65 may be a
compound of formula 7a wherein Z is O.
[0603] In certain embodiments, an inhibitor of RPE65 may be a
compound of formula 7b:
##STR00110## [0604] wherein, independently for each occurrence:
[0605] Y is O, S, NR', CH.sub.2.dbd.O, C.dbd.S, C.dbd.NR', CHOR',
CHNR'R'', CHSR', or CH.sub.2; [0606] R.sub.1 is R', --OR', --CN or
(CH.sub.2CH.sub.2O).sub.mR'; [0607] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl; [0608] R'' is H,
alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or
heteroaralkyl;
[0608] ##STR00111## [0609] m is 1, 2 or 3; and [0610] n is 1, 2, or
3; [0611] wherein denotes a single bond, a cis double bond or a
trans double bond.
[0612] Compounds of formula 7b may be considered reversible
antagonists of RPE65 because they can noncovalently bind RPE65
without permanently disabling it.
[0613] In certain embodiments, an inhibitor of RPE65 may be a
compound of formula 7e:
##STR00112## [0614] wherein, independently for each occurrence:
[0615] X is O, S, NR', CH.sub.2, or NHNR'; [0616] Z is O or
NOH;
[0617] R.sub.1 is --CH.sub.2F, --CHF.sub.2, --CF.sub.3,
--CH.sub.2N.sub.2, --CH.sub.2C(O)OR', --OR', --C(O)CHR',
--C(NH)CHR', or --CH.dbd.CHR'; [0618] R' is H, alkyl, heteroalkyl,
aryl, heteroaryl, aralkyl, or heteroaralkyl;
[0618] ##STR00113## [0619] n is 1, 2, or 3.
[0620] Compounds of formula 7c may be considered irreversible
antagonists of RPE65 because they can covalently bind RPE65,
permanently disabling it.
[0621] In certain embodiments, an inhibitor of RPE65 may be a
compound of formula 7c wherein Z is O.
[0622] In certain embodiments, an inhibitor of RPE65 may be a
compound of formula 7d:
##STR00114## [0623] wherein, independently for each occurrence:
[0624] Y is C.dbd.O, C.dbd.S, C.dbd.NR', CHOH, CHOR', NH.sub.2,
NHR', NR'R'', SH, SR', or CH.sub.2; [0625] R.sub.1 is R', --OR',
--CN or --(CH.sub.2CH.sub.2O).sub.mR'; [0626] R' is H, alkyl,
heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl; [0627]
R'' is H, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or
heteroaralkyl;
[0628] R is
##STR00115## [0629] m is 1, 2 or 3; and [0630] n is 1, 2, or 3.
[0631] B. Compositions for Short-Circuiting
[0632] Short-circuiting the visual cycle can be achieved by
catalyzing the thermodynamically downhill isomerization of
11-cis-retinal to all-trans-retinal in the RPE, before the
11-cis-retinal leaves the RPE. FIG. 3 depicts one contemplated
intervention. A very wide variety of substances are envisioned as
appropriate for this use. Broadly speaking, appropriate drugs
include aniline derivates, i.e., a benzene ring with an amine side
chain.
[0633] Short circuiting molecules operate by first forming a Schiff
base with a retinal. When a Schiff base is formed with
11-cis-retinal, isomerization occurs. This is the short
circuit.
[0634] Short-circuit compounds may also trap retinals so that they
are not available to form A.sub.2E, its precursors or analogs. With
all-trans-retinal, a relatively stable Schiff base can be formed
with the drugs which traps the all-trans-retinal and prevents it
from forming A.sub.2E and like compounds. The short-circuit drug
competes with phosphatidylethanolamine for binding
all-trans-retinal. The trapped compounds may then be broken down in
lysozomes to non-toxic metabolites. A short-circuit drug may
disrupt the visual cycle in one or both ways, i.e., by
short-circuiting 11-cis-retinals and/or by trapping
all-trans-retinals. (A.sub.2E is the best characterized of the
lipofuscins. There may be other adducts between all-trans-retinal
and amines--or even proteins--whose formation is initiated by
Schiff base formation between a reactive retinal and an amine.)
[0635] While it is not expected that an aromatic
amine/all-trans-retinal Schiff base will go on to form
A.sub.2E-like molecules (because it will be degraded first), this
can be more reliably prevented by using a short-circuiting drug
that is a secondary amine. This is because the mechanism of
A.sub.2E formation requires a primary amine (two free Hs) because
two new N-alkyl bonds are made (one with each all-trans-retinal
molecule) and this cannot happen starting with a secondary or
tertiary amine. If the short-circuit drug is a secondary amine,
then it can bind only one molecule of all-trans-retinal and has no
remaining site to bind a second all-trans-retinal, thereby
preventing the formation of compounds analogous to A.sub.2E akin to
the process shown in FIG. 2.
[0636] Short-circuit drugs may also provide a long-term effect, so
that their administration can be infrequent. In some cases,
administration may be required monthly. In other cases,
administration may be required weekly. The short-circuit drugs
effectively deplete vitamin A stores locally in the eye by trapping
all-trans-retinal. Once the store of vitamin is diminished by the
drug, the visual cycle is impaired, and lipofuscin formation is
retarded, which is the goal of therapy. Vitamin A stores are
replenished only very slowly in the eye, so that a single
administration of short-circuit drug may have a prolonged effect.
In addition, the short-circuit drugs may be cleared slowly from the
eye, so that they may be available for binding over extended
periods.
[0637] In certain embodiments, a short-circuiting compound has the
structure represented by formula VII:
##STR00116## [0638] wherein, independently for each occurrence:
[0639] R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, or carbonyl; [0640] L is a hydrophobic moiety, or
any two adjacent L taken together form a fused aromatic or
heteroaromatic ring (e.g. a naphthalene, an anthracene, an indole,
a quinoline, etc.).
[0641] In certain embodiments, independently for each occurrence, L
is alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, carbonyl, ether, or polycyclic. In certain
embodiments, L has the formula VIa:
##STR00117## [0642] wherein, independently for each occurrence:
[0643] R' and X are hydrogen, alkyl, alkenyl, alkynyl, aryl,
aralkyl, heteroaryl, heteroaralkyl, carbonyl, alkoxy, hydroxy,
thiol, thioalkyl, or amino; and [0644] m is an integer from 1 to 6
inclusive.
[0645] In some embodiments, a short circuit drug may be represented
by the following generic formula VIb:
##STR00118##
wherein n is an integer from 1 to 8 inclusive.
[0646] In some embodiments, a short circuit drug may be represented
by the following generic formula VIc:
##STR00119##
[0647] wherein, independently for each occurrence,
[0648] R is H, alkyl, or acyl; and
[0649] R' is alkyl or ether.
[0650] In a further embodiment, a short circuit drug has the
structure of formula VIIc and the attendant definitions, wherein R
is H for both occurrences.
[0651] In a further embodiment, a short circuit drug has the
structure of formula VIIc and the attendant definitions, wherein at
least one R is alkyl.
[0652] In a further embodiment, a short circuit drug has the
structure of formula VIIc and the attendant definitions, wherein at
least one R is methyl.
[0653] In some embodiments, a short circuit drug may be represented
by the following generic formula VIId:
##STR00120##
[0654] wherein, independently for each occurrence:
[0655] R is H, alkyl, or acyl; and
[0656] R' is alkyl or ether.
[0657] In a further embodiment, a short circuit drug has the
structure of formula VIId and the attendant definitions, wherein R
is H for both occurrences.
[0658] In a further embodiment, a short circuit drug has the
structure of formula VIId and the attendant definitions, wherein at
least one R is alkyl.
[0659] In a further embodiment, a short circuit drug has the
structure of formula VIId and the attendant definitions, wherein at
least one R is methyl.
[0660] In some embodiments, a short circuit drug may be represented
by the following generic formula VIIe:
##STR00121##
[0661] wherein, independently for each occurrence:
[0662] X is hydrogen or --C(.dbd.O)OR';
[0663] R is H, alkyl, or acyl; and
[0664] R' is alkyl.
[0665] In a further embodiment, a short circuit drug has the
structure of formula VIIe and the attendant definitions, wherein R
is H.
[0666] In a further embodiment, a short circuit drug has the
structure of formula VIIe and the attendant definitions, wherein at
least one R is alkyl.
[0667] In a further embodiment, a short circuit drug has the
structure of formula VIIe and the attendant definitions, wherein R
is methyl.
[0668] In some embodiments, a short circuit drug may be represented
by the following generic formula VIIf:
##STR00122##
[0669] wherein, independently for each occurrence
[0670] R is H, alkyl, or acyl; and
[0671] R' is alkyl.
[0672] In a further embodiment, a short circuit drug has the
structure of formula VIIf and the attendant definitions, wherein R
is H.
[0673] In a further embodiment, a short circuit drug has the
structure of formula VIIf and the attendant definitions, wherein at
least one R is alkyl.
[0674] In a further embodiment, a short circuit drug has the
structure of formula VIIf and the attendant definitions, wherein R
is methyl.
[0675] In one embodiment, a short circuiting drug is
diaminophenoxypentane:
##STR00123##
[0676] In one embodiment, a short circuiting drug is
phenetidine:
##STR00124##
[0677] In one embodiment, a short circuiting drug is and
tricaine:
##STR00125##
[0678] In one embodiment, a short circuiting drug is
4-butylanaline:
##STR00126##
[0679] In one embodiment, a short circuiting drug is
N-methyl-4-butylanaline:
##STR00127##
[0680] In one embodiment, a short circuiting drug is ethyl
3-aminobenzoate:
##STR00128##
[0681] In one embodiment, a short circuiting drug is ethyl
N-methyl-3-aminobenzoate:
##STR00129##
[0682] In one embodiment, a short circuiting drug is ethyl
2-aminobenzoate:
##STR00130##
[0683] In one embodiment, a short circuiting drug is ethyl
N-methyl-2-aminobenzoate:
##STR00131##
[0684] In some embodiments, a short circuit drug may be represented
by the following generic formula VIII:
##STR00132## [0685] wherein R.sup.1 is hydrogen, alkyl or ether; or
any two adjacent L taken together form a fused aromatic or
heteroaromatic ring (e.g. a naphthalene, an anthracene, etc.).
[0686] In certain embodiments, a short-circuiting compound has the
structure represented by formula IX:
ANR.sub.2 IX [0687] wherein, independently for each occurrence:
[0688] R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, or carbonyl; and [0689] A is optionally substituted
aryl or heteroaryl.
[0690] In some embodiments, a short circuit drug may be represented
by the following generic formula X:
AC(.dbd.O)NHNH.sub.2 X
[0691] wherein independently for each occurrence:
[0692] R' is hydrogen, alkyl or ether; and
[0693] A is optionally substituted aryl or heteroaryl.
[0694] In certain embodiments, a short-circuiting compound may have
a structure represented by general structure 5:
##STR00133## [0695] wherein, independently for each occurrence:
[0696] R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, or carbonyl;
[0697] L is a hydrophobic moiety, or any two adjacent L taken
together form a fused aromatic ring; and [0698] n is an integer
from 0 to 5 inclusive.
[0699] In certain embodiments, independently for each occurrence, L
is alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, carbonyl, ether, or polycyclic. In certain
embodiments, L has the formula 5a:
##STR00134## [0700] wherein, independently for each occurrence:
[0701] R' and X are H, alkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, heteroaralkyl, carbonyl, alkoxy, hydroxy, thiol,
thioalkyl, or amino; [0702] m is an integer from 1 to 6 inclusive;
and [0703] p is an integer from 0 to 5 inclusive.
[0704] Selected specific examples of short circuit drugs include
diaminophenoxypentane:
##STR00135##
[0705] In some embodiments, a short circuit drug may be represented
by the following generic formula 5b:
##STR00136## [0706] wherein n is an integer from 1 to 8
inclusive.
[0707] In some embodiments, a short circuit drug may be represented
by the following generic formula 5c:
##STR00137## [0708] wherein, independently for each occurrence:
[0709] R is H, alkyl, or acyl; and [0710] R' is alkyl or ether.
[0711] In a further embodiment, a short circuit drug has the
structure of formula 5c and the attendant definitions, wherein R is
H for both occurrences.
[0712] In a further embodiment, a short circuit drug has the
structure of formula 5c and the attendant definitions, wherein at
least one R is alkyl.
[0713] In a further embodiment, a short circuit drug has the
structure of formula 5c and the attendant definitions, wherein at
least one R is methyl.
[0714] In some embodiments, a short circuit drug may be represented
by the following generic formula 5c1:
##STR00138## [0715] wherein, independently for each occurrence:
[0716] R is H, alkyl, or acyl; and [0717] R' is alkyl or ether.
[0718] In a further embodiment, a short circuit drug has the
structure of formula 5 .mu.l and the attendant definitions, wherein
R is H for both occurrences.
[0719] In a further embodiment, a short circuit drug has the
structure of formula 5 .mu.l and the attendant definitions, wherein
at least one R is alkyl.
[0720] In a further embodiment, a short circuit drug has the
structure of formula 5 .mu.l and the attendant definitions, wherein
at least one R is methyl.
[0721] In some embodiments, a short circuit drug may be represented
by the following generic formula 5d:
##STR00139## [0722] wherein, independently for each occurrence:
[0723] R is H, alkyl, or acyl; and [0724] R' is alkyl.
[0725] In a further embodiment, a short circuit drug has the
structure of formula 5d1 and the attendant definitions, wherein R
is H.
[0726] In a further embodiment, a short circuit drug has the
structure of formula 5d and the attendant definitions, wherein at
least one R is alkyl.
[0727] In a further embodiment, a short circuit drug has the
structure of formula 5d and the attendant definitions, wherein R is
methyl.
[0728] In some embodiments, a short circuit drug may be represented
by the following generic formula 5d1:
##STR00140## [0729] wherein, independently for each occurrence:
[0730] R is H, alkyl, or acyl; and [0731] R' is alkyl.
[0732] In a further embodiment, a short circuit drug has the
structure of formula 5d1 and the attendant definitions, wherein R
is H.
[0733] In a further embodiment, a short circuit drug has the
structure of formula 5d1 and the attendant definitions, wherein at
least one R is alkyl.
[0734] In a further embodiment, a short circuit drug has the
structure of formula 5d1 and the attendant definitions, wherein R
is methyl.
[0735] In some embodiments, a short circuit drug may be represented
by the following generic formula 5e:
##STR00141## [0736] wherein R' is alkyl or ether.
[0737] Diseases associated with lipofuscin accumulation may also be
treated or prevented with agents or drugs that prevent vitamin A
import into the eye. Exemplary agents are those that prevent
vitamin A delivery by retinol binding protein (RBP). Agents may
thus be RBP inhibitors or blocking agents. RBP may be a CRBP
protein (Ong et al. (1994) Nutr. Rev. 52:524 and Cowan et al.
(1993) J. Mol. Biol. 230:1225) or a serum RBP (Blomhoff et al.
(1990) Science 250:399 and Newcomer et al. (1984) EMBO J. 3, 1451).
A preferred RBP to inhibit is CRBP-1. In an illustrative
embodiment, the RBP blocking agent is fenretinide, a non-retinoid
fenretinide analog or a non-retinoid fenretinide isoprenoid.
Exemplary compounds have the structure depicted in formula XI:
##STR00142## [0738] wherein, independently for each occurrence,
[0739] n is 0 to 10 inclusive; [0740] R.sup.1 is hydrogen or alkyl;
[0741] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; [0742] Z is hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl,
--C(.dbd.O)R.sub.b, or --(CH.sub.2).sub.pR.sub.b; [0743] p is 0 to
20 inclusive; [0744] R.sub.a is hydrogen, alkyl, cycloalkyl,
alkenyl, cycloalkenyl, alkynyl, aryl, or aralkyl; [0745] R.sub.b is
hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,
or aralkyl; and [0746] denotes a single bond or a trans double
bond.
[0747] In a further embodiment a RBP blocking agent has the
structure of XI, wherein R.sup.1 is hydrogen or methyl.
[0748] In a further embodiment a RBP blocking agent has the
structure of XI, wherein Z is aryl.
[0749] In a further embodiment a RBP blocking agent has the
structure of XI, wherein R.sub.a is hydrogen.
[0750] In a further embodiment a RBP blocking agent has the
structure of XI, wherein R.sup.1 is hydrogen or methyl; Z is aryl;
and R.sub.a is hydrogen.
[0751] In a another embodiment a RBP blocking agent of the
invention has the structure of formula XIa:
##STR00143## [0752] wherein, independently for each occurrence,
[0753] n is 0 to 10 inclusive; [0754] R.sup.1 is hydrogen or alkyl;
[0755] R.sup.2 is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, or aralkyl; [0756] R.sup.3, R.sup.4,
R.sup.5, R.sup.6 and R.sup.7 are hydrogen, halogen, alkyl, alkenyl,
alkynyl, aryl, heteroaryl, aralkyl, aralkyenyl, aralkynyl,
heteroaralkyl, heteroaralkyenyl, heteroaralkynyl, cyano, nitro,
sulfhydryl, hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio,
carboxyl, carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido,
sulfamoyl, sulfonyl, or sulfoxido; [0757] R.sub.a is hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, or
aralkyl; and [0758] denotes a single bond or a trans double
bond.
[0759] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sup.1 is hydrogen or methyl.
[0760] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sub.a is hydrogen.
[0761] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sup.1 is hydrogen or methyl; and
R.sub.a is hydrogen.
[0762] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sup.3, R.sup.4, R.sup.6 and R.sup.7 are
hydrogen.
[0763] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sup.5 is hydroxyl.
[0764] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sup.3, R.sup.4, R.sup.6 and R.sup.7 are
hydrogen; and R.sup.5 is hydroxyl.
[0765] In a further embodiment a RBP blocking agent has the
structure of XIa, wherein R.sup.1 is hydrogen or methyl; R.sub.a is
hydrogen; R.sup.3, R.sup.4, R.sup.6 and R.sup.7 are hydrogen; and
R.sup.5 is hydroxyl.
[0766] In a another embodiment a RBP blocking agent of the
invention has the structure of formula XIb:
##STR00144## [0767] wherein, independently for each occurrence,
[0768] n is 0 to 5 inclusive; [0769] R.sup.1 is hydrogen or methyl;
[0770] R.sup.2 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
[0770] ##STR00145## [0771] R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7 and R.sup.8 are hydrogen, halogen, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, aralkyl, aralkyenyl, aralkynyl, heteroaralkyl,
heteroaralkyenyl, heteroaralkynyl, cyano, nitro, sulfhydryl,
hydroxyl, sulfonyl, amino, acylamino, amido, alkylthio, carboxyl,
carbamoyl, alkoxyl, sulfonate, sulfate, sulfonamido, sulfamoyl,
sulfonyl, or sulfoxido; [0772] any two geminal R.sup.8 and the
carbon to which they are bound may represent C(.dbd.O); and [0773]
R.sub.a is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, aryl, or aralkyl.
[0774] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.1 is hydrogen or methyl.
[0775] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sub.a is hydrogen.
[0776] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.1 is hydrogen or methyl; and
R.sub.a is hydrogen.
[0777] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.3, R.sup.4, R.sup.6 and R.sup.7 are
hydrogen.
[0778] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.5 is hydroxyl.
[0779] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.3, R.sup.4, R.sup.6 and R.sup.7 are
hydrogen; and R.sup.5 is hydroxyl.
[0780] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.1 is hydrogen or methyl; R.sub.a is
hydrogen; R.sup.3, R.sup.4, R.sup.6 and R.sup.7 are hydrogen; and
R.sup.5 is hydroxyl.
[0781] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein n is 1 to 3 inclusive.
[0782] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.2 is
##STR00146##
[0783] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.2 is
##STR00147##
[0784] In a further embodiment a RBP blocking agent has the
structure of XIb, wherein R.sup.2 is
##STR00148##
[0785] An RBP inhibitory compound may be a non-retinoid fenretinide
analog, such as those set forth above, wherein the analog is not
fenretinide.
[0786] Vitamin A delivery to the eye may also be inhibited by
interfering with the membrane receptor for holo-RBP, which have
been reported to be present in the RBP (Vogel et al. (1999) cited
in Quadro et al. (1999) EMBO J. 18:4633). Alternatively, the
interaction between RBP and its receptor may be inhibited.
[0787] In yet other embodiments, inhibitors of retinyl-ester
isomerase (e.g., 11-cis-retinyl bromoacetate) and/or inhibitors of
cellular retinaldehyde binding protein (CRALBP) (e.g.,
cis-isoprenoids) and/or inhibitors of 11-cis-retinol dehydrogenase
(e.g., pyrazoles) may be used.
[0788] Also included are pharmaceutically acceptable addition salts
and complexes of the compounds of the formulas given above. In
cases wherein the compounds may have one or more chiral centers,
unless specified, the compounds contemplated herein may be a single
stereoisomer or racemic mixtures of stereoisomers. Further included
are prodrugs, analogs, and derivatives thereof.
[0789] In some embodiments, a combination of one or more compounds
described herein is administered to a subject. Two or more
short-circuiting compounds may be combined. In some embodiments, an
enzyme inhibitor and/or RPE65 binding inhibitor may be combined
with a short-circuiting compound. An enzyme inhibitor and/or
short-circuiting compound may also be combined with one or more
agents that prevent the delivery of vitamin A to the eye, such as
fenretinide, a non-retinoid fenretinide analog and a non-retinoid
isoprenoid. For example, a compound of formula XI may be
administered to a subject who is also receiving a compound of
formula I, II, III, IV, V, VI, VII, VIII, IX, or X. Any other
combination of compounds affecting different target proteins may
also be used.
[0790] A compound described herein, e.g., a compound of formula XI,
may be administered to a subject who is also receiving a compound
selected from the group consisting of C-glycoside and arylamide
analogues of N-(4-hydroxyphenyl)retinamide-O-glucuronide, including
but not limited to 4-(retinamido)phenyl-C-glucuronide,
4-(retinamido)phenyl-C-glucoside, 4-(retinamido)phenyl-C-xyloside,
4-(retinamido)benzyl-C-glucuronide,
4-(retinamido)benzyl-C-glucoside, 4-(retinamido)benzyl-C-xyloside;
and retinoyl .beta.-glucuronide analogues such as, for example,
1-(.beta.-D-glucopyranosyl) retinamide and
1-(D-glucopyranosyluronosyl)retinamide, described in U.S. Pat. Nos.
5,516,792, 5,663,377, 5,599,953, 5,574,177, and Bhatnagar et al,
Biochem. Pharmacol., 41:1471-7 (1991), each incorporated herein by
reference. Other fenretinide derivatives, such as those described
in WO2006/063128 and WO2006/007314, may be contemplated for use in
certain embodiments. Additional compounds which may be used in
combination with the inventive compounds, e.g., a compound of
formula XI, are also disclosed in WO2006/063128 and WO2006/007314.
Both WO2006/063128 and WO2006/007314 are incorporated herein by
reference in their entirety.
[0791] In certain embodiments, a compound described herein, e.g., a
compound of formula XI, may be administered to a subject who is
also receiving a vitamin A derivative, including those disclosed in
U.S. Pat. No. 4,743,400, incorporated herein by reference. These
retinoids include, for example, all-trans retinoyl chloride,
all-trans-4-(methoxyphenyl) retinamide (methoxyphenyl retinamide),
13-cis-4-(hydroxyphenyl) retinamide and all-trans-4-(ethoxyphenyl)
retinamide. U.S. Pat. No. 4,310,546, incorporated herein by
reference, describes N-(4-acyloxyphenyl)-all-trans retinamides,
such as, for example, N-(4-acetoxyphenyl)-all-trans-retinamide,
N-(4-propionyloxyphenyl)-all-trans-retinamide and
N-(4-N-butyryloxyphenyl)-all-trans-retinamide, all of which are
contemplated for use in certain embodiments. Other vitamin A
derivatives or metabolites, such as N-(1H-tetrazol-5-yl)retinamide,
N-ethylretinamide, 13-cis-N-ethylretinamide, N-butylretinamide,
etretin (acitretin), etretinate, tretinoin (all-trans-retinoic
acid) or isotretinoin (13-cis-retinoic acid) may be contemplated
for use in certain embodiments. See U.S. Provisional Patent
Applications Nos. 60/582,293 and 60/602,675; see also Turton et
al., Int. J. Exp. Pathol., 73:551-63 (1992), all herein
incorporated by reference.
[0792] In combination therapies, the compounds may be administered
simultaneously, e.g., in the form of one composition, or
consecutively. When consecutively administered, the time between
the two administrations may be one or more minutes, one or more
hours, one or more days, or one or more weeks.
[0793] Therapeutic or prophylactic treatments with one or more of
the compounds described herein may be combined with other
treatments known in the art. For example, they may be used in
conjunction with surgery and/or supplements, and/or radiation,
and/or other therapeutic methods. Treatment of the wet form of AMD
may be combined a treatment that removes or destroys the new blood
vessels that grow in or around the macula. A laser, such as a
thermal laser may be used for that purpose. Transpupillary
thermotherapy is an alternative treatment, in which an infrared
laser is used. Another method is photodynamic therapy, in which a
substance that sensitizes the blood vessels in the eye to laser
light is given intravenously, and then a beam of laser light is
used to destroy the abnormal blood vessels. Photocoagulation
therapy may also be used.
[0794] Treatments may also be combined with administration of an
anti-oxidant, e.g., high doses of antioxidants, such as (vitamin C,
vitamin E, and beta-carotene), zinc and copper ("supplementation
therapy"). The antioxidant formulation may contain a combination of
vitamin C, vitamin E, and beta-carotene. The specific daily amounts
of antioxidants and zinc may be about 500 milligrams of vitamin C;
400 international units of vitamin E; 15 milligrams of
beta-carotene; 80 milligrams of zinc as zinc oxide; and two
milligrams of copper as cupric oxide (used in the Age-Related Eye
Disease (ARED) study). A number of new drugs have promise for the
prevention or delay of photoreceptor cell death and retinal
degeneration. These drugs include PKC 412 (which blocks chemicals
in the body that foster new blood vessel growth, or angiogenesis),
Glial Derived Neurotrophic Factor (a survival factor which has
slowed degeneration in a rodent model), and diatazem (a
calcium-channel blocker which addresses a rare retinal gene defect
called beta PDE).
[0795] Where a treatment of macular degeneration described herein
is combined with radiotherapy, preferred forms of radiation for use
in the treatment include: proton beam, strontium-90, palladium-103,
radiosurgery, and EBRT (external beam radiation therapy). Radiation
therapy for "wet" macular degeneration is used to destroy blood
vessels and prevent neovascularization. Radiation therapy is useful
after surgery to prevent or reduce scarring by killing or effecting
the cells which make up newly formed blood vessels, inflammatory
cells which promote scarring and cells which help create fibrous
tissues.
[0796] The two therapies may be provided simultaneously or
consecutively. For example, a surgical method may be applied first,
followed by administration of one or more compounds described
herein. Alternatively, a surgical method may be used after
administration of one or more compounds described herein. The
second therapeutic method, such as surgery, may also be preceded by
and followed by administration of one or more compounds described
herein.
[0797] The other therapy may precede or follow the therapeutic
agent-based therapy by intervals ranging from minutes to days to
weeks. In embodiments where the other macular or retinal
degeneration therapy and the therapeutic agent-based therapy are
administered together, one may prefer to avoid that a significant
period of time did not expire between the time of each delivery. In
such instances, it is contemplated that one would administer to a
patient both modalities within about 12-24 hours of each other and,
more preferably, within about 6-12 hours of each other, with a
delay time of only about 12 hours being most preferred. In some
situations, it may be desirable to extend the time period for
treatment significantly, however, where several days (2, 3, 4, 5, 6
or 7) to several weeks (1, 2, 3, 4, 5, 6, 7 or 8) lapse between the
respective administrations.
[0798] It also is conceivable that more than one administration of
either the other macular or retinal degeneration therapy and the
therapeutic agent-based therapy will be required to prevent
blindness or a decrease in vision. Various combinations may be
employed, where the other macular or retinal degeneration therapy
is "A" and the therapeutic agent-based therapy treatment is "B", as
exemplified below:
TABLE-US-00001 A/B/A B/A/B B/B/A A/A/B B/A/A A/B/B B/B/B/A B/B/A/B
A/A/B/B A/B/A/B A/B/B/A B/B/A/A B/A/B/A B/A/A/B B/B/B/A A/A/A/B
B/A/A/A A/B/A/A A/A/B/A A/B/B/B B/A/B/B B/B/A/B.
[0799] Pharmaceutical compositions for use in accordance with the
present methods may be formulated in conventional manner using one
or more physiologically acceptable carriers or excipients. Thus,
activating compounds and their physiologically acceptable salts and
solvates may be formulated for administration by, for example,
injection, inhalation or insufflation (either through the mouth or
the nose) or oral, buccal, parenteral or rectal administration. In
one embodiment, the compound is administered locally, at the site
where the target cells, e.g., diseased cells, are present, i.e., in
the eye or the retina.
[0800] Compounds can be formulated for a variety of loads of
administration, including systemic and topical or localized
administration. Techniques and formulations generally may be found
in Remington's Pharmaceutical Sciences, Meade Publishing Co.,
Easton, Pa. For systemic administration, injection is preferred,
including intramuscular, intravenous, intraperitoneal, and
subcutaneous. For injection, the compounds can be formulated in
liquid solutions, preferably in physiologically compatible buffers
such as Hank's solution or Ringer's solution. In addition, the
compounds may be formulated in solid form and redissolved or
suspended immediately prior to use. Lyophilized forms are also
included.
[0801] For oral administration, the pharmaceutical compositions may
tale the form of, for example, tablets, lozanges, or capsules
prepared by conventional means with pharmaceutically acceptable
excipients such as binding agents (e.g., pregelatinised maize
starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose);
fillers (e.g., lactose, microcrystalline cellulose or calcium
hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or
silica); disintegrants (e.g., potato starch or sodium starch
glycolate); or wetting agents (e.g., sodium lauryl sulphate). The
tablets may be coated by methods well known in the art. Liquid
preparations for oral administration may take the form of, for
example, solutions, syrups or suspensions, or they may be presented
as a dry product for constitution with water or other suitable
vehicle before use. Such liquid preparations may be prepared by
conventional means with pharmaceutically acceptable additives such
as suspending agents (e.g., sorbitol syrup, cellulose derivatives
or hydrogenated edible fats); emulsifying agents (e.g., lecithin or
acacia); non-aqueous vehicles (e.g., ationd oil, oily esters, ethyl
alcohol or fractionated vegetable oils); and preservatives (e.g.,
methyl or propyl-p-hydroxybenzoates or sorbic acid). The
preparations may also contain buffer salts, flavoring, coloring and
sweetening agents as appropriate. Preparations for oral
administration may be suitably formulated to give controlled
release of the active compound.
[0802] For administration by inhalation, the compounds may be
conveniently delivered in the form of an aerosol spray presentation
from pressurized packs or a nebulizer, with the use of a suitable
propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In
the case of a pressurized aerosol the dosage unit may be determined
by providing a valve to deliver a metered amount. Capsules and
cartridges of e.g., gelatin, for use in an inhaler or insufflator
may be formulated containing a powder mix of the compound and a
suitable powder base such as lactose or starch.
[0803] The compounds may be formulated for parenteral
administration by injection, e.g., by bolus injection or continuous
infusion. Formulations for injection may be presented in unit
dosage form, e.g., in ampoules or in multi-dose containers, with an
added preservative. The compositions may take such forms as
suspensions, solutions or emulsions in oily or aqueous vehicles,
and may contain formulatory agents such as suspending, stabilizing
and/or dispersing agents. Alternatively, the active ingredient may
be in powder form for constitution with a suitable vehicle, e.g.,
sterile pyrogen-free water, before use.
[0804] The compounds may also be formulated in rectal compositions
such as suppositories or retention enemas, e.g., containing
conventional suppository bases such as cocoa butter or other
glycerides.
[0805] In addition to the formulations described previously, the
compounds may also be formulated as a depot preparation. Such long
acting formulations may be administered by implantation (for
example subcutaneously or intramuscularly) or by intramuscular
injection. Thus, for example, the compounds may be formulated with
suitable polymeric or hydrophobic materials (for example as an
emulsion in an acceptable oil) or ion exchange resins, or as
sparingly soluble derivatives, for example, as a sparingly soluble
salt.
[0806] Pharmaceutical compositions (including cosmetic
preparations) may comprise from about 0.00001 to 100% such as from
0.001 to 10% or from 0.1% to 5% by weight of one or more compounds
described herein.
[0807] In one embodiment, a compound described herein, is
incorporated into a topical formulation containing a topical
carrier that is generally suited to topical drug administration and
comprising any such material known in the art. The topical carrier
may be selected so as to provide the composition in the desired
form, e.g., as an ointment, lotion, cream, microemulsion, gel, oil,
solution, or the like, and may be comprised of a material of either
naturally occurring or synthetic origin. It is preferable that the
selected carrier not adversely affect the active agent or other
components of the topical formulation. Examples of suitable topical
carriers for use herein include water, alcohols and other nontoxic
organic solvents, glycerin, mineral oil, silicone, petroleum jelly,
lanolin, fatty acids, vegetable oils, parabens, waxes, and the
like.
[0808] Formulations may be colorless, odorless ointments, lotions,
creams, microemulsions and gels.
[0809] Compounds may be incorporated into ointments, which
generally are semisolid preparations which are typically based on
petrolatum or other petroleum derivatives. The specific ointment
base to be used, as will be appreciated by those skilled in the
art, is one that will provide for optimum drug delivery, and,
preferably, will provide for other desired characteristics as well,
e.g., emolliency or the like. As with other carriers or vehicles,
an ointment base should be inert, stable, nonirritating and
nonsensitizing. As explained in Remington's, cited in the preceding
section, ointment bases may be grouped in four classes: oleaginous
bases; emulsifiable bases; emulsion bases; and water-soluble bases.
Oleaginous ointment bases include, for example, vegetable oils,
fats obtained from animals, and semisolid hydrocarbons obtained
from petroleum. Emulsifiable ointment bases, also known as
absorbent ointment bases, contain little or no water and include,
for example, hydroxystearin sulfate, anhydrous lanolin and
hydrophilic petrolatum. Emulsion ointment bases are either
water-in-oil (W/O) emulsions or oil-in-water (O/W) emulsions, and
include, for example, cetyl alcohol, glyceryl monostearate, lanolin
and stearic acid. Exemplary water-soluble ointment bases are
prepared from polyethylene glycols (PEGs) of varying molecular
weight; again, reference may be had to Remington's, supra, for
further information.
[0810] Compounds may be incorporated into lotions, which generally
are preparations to be applied to the skin surface without
friction, and are typically liquid or semiliquid preparations in
which solid particles, including the active agent, are present in a
water or alcohol base. Lotions are usually suspensions of solids,
and may comprise a liquid oily emulsion of the oil-in-water type.
Lotions are preferred formulations for treating large body areas,
because of the ease of applying a more fluid composition. It is
generally necessary that the insoluble matter in a lotion be finely
divided. Lotions will typically contain suspending agents to
produce better dispersions as well as compounds useful for
localizing and holding the active agent in contact with the skin,
e.g., methylcellulose, sodium carboxymethylcellulose, or the like.
An exemplary lotion formulation for use in conjunction with the
present method contains propylene glycol mixed with a hydrophilic
petrolatum such as that which may be obtained under the trademark
Aquaphor.RTM. from Beiersdorf, Inc. (Norwalk, Conn.).
[0811] Compounds may be incorporated into creams, which generally
are viscous liquid or semisolid emulsions, either oil-in-water or
water-in-oil. Cream bases are water-washable, and contain an oil
phase, an emulsifier and an aqueous phase. The oil phase is
generally comprised of petrolatum and a fatty alcohol such as cetyl
or stearyl alcohol; the aqueous phase usually, although not
necessarily, exceeds the oil phase in volume, and generally
contains a humectant. The emulsifier in a cream formulation, as
explained in Remington's, supra, is generally a nonionic, anionic,
cationic or amphoteric surfactant.
[0812] Compounds may be incorporated into microemulsions, which
generally are thermodynamically stable, isotropically clear
dispersions of two immiscible liquids, such as oil and water,
stabilized by an interfacial film of surfactant molecules
(Encyclopedia of Pharmaceutical Technology (New York: Marcel
Dekker, 1992), volume 9). For the preparation of microemulsions,
surfactant (emulsifier), co-surfactant (co-emulsifier), an oil
phase and a water phase are necessary. Suitable surfactants include
any surfactants that are useful in the preparation of emulsions,
e.g., emulsifiers that are typically used in the preparation of
creams. The co-surfactant (or "co-emulsifer") is generally selected
from the group of polyglycerol derivatives, glycerol derivatives
and fatty alcohols. Preferred emulsifier/co-emulsifier combinations
are generally although not necessarily selected from the group
consisting of: glyceryl monostearate and polyoxyethylene stearate;
polyethylene glycol and ethylene glycol palmitostearate; and
caprilic and capric triglycerides and oleoyl macrogolglycerides.
The water phase includes not only water but also, typically,
buffers, glucose, propylene glycol, polyethylene glycols,
preferably lower molecular weight polyethylene glycols (e.g., PEG
300 and PEG 400), and/or glycerol, and the like, while the oil
phase will generally comprise, for example, fatty acid esters,
modified vegetable oils, silicone oils, mixtures of mono- di- and
triglycerides, mono- and di-esters of PEG (e.g., oleoyl macrogol
glycerides), etc.
[0813] Compounds may be incorporated into gel formulations, which
generally are semisolid systems consisting of either suspensions
made up of small inorganic particles (two-phase systems) or large
organic molecules distributed substantially uniformly throughout a
carrier liquid (single phase gels). Single phase gels can be made,
for example, by combining the active agent, a carrier liquid and a
suitable gelling agent such as tragacanth (at 2 to 5%), sodium
alginate (at 2-10%), gelatin (at 2-15%), methylcellulose (at 3-5%),
sodium carboxymethylcellulose (at 2-5%), carbomer (at 0.3-5%) or
polyvinyl alcohol (at 10-20%) together and mixing until a
characteristic semisolid product is produced. Other suitable
gelling agents include methylhydroxycellulose,
polyoxyethylene-polyoxypropylene, hydroxyethylcellulose and
gelatin. Although gels commonly employ aqueous carrier liquid,
alcohols and oils can be used as the carrier liquid as well.
[0814] Various additives, known to those skilled in the art, may be
included in formulations, e.g., topical formulations. Examples of
additives include, but are not limited to, solubilizers, skin
permeation enhancers, opacifiers, preservatives (e.g.,
anti-oxidants), gelling agents, buffering agents, surfactants
(particularly nonionic and amphoteric surfactants), emulsifiers,
emollients, thickening agents, stabilizers, humectants, colorants,
fragrance, and the like. Inclusion of solubilizers and/or skin
permeation enhancers is particularly preferred, along with
emulsifiers, emollients and preservatives. An optimum topical
formulation comprises approximately: 2 wt. % to 60 wt. %,
preferably 2 wt. % to 50 wt. %, solubilizer and/or skin permeation
enhancer; 2 wt. % to 50 wt. %, preferably 2 wt. % to 20 wt. %,
emulsifiers; 2 wt. % to 20 wt. % emollient; and 0.01 to 0.2 wt. %
preservative, with the active agent and carrier (e.g., water)
making of the remainder of the formulation.
[0815] A skin permeation enhancer serves to facilitate passage of
therapeutic levels of active agent to pass through a reasonably
sized area of unbroken skin. Suitable enhancers are well known in
the art and include, for example: lower alkanols such as methanol
ethanol and 2-propanol; alkyl methyl sulfoxides such as
dimethylsulfoxide (DMSO), decylmethylsulfoxide (C.sub.10 MSO) and
tetradecylmethyl sulfoxide; pyrrolidones such as 2-pyrrolidone,
N-methyl-2-pyrrolidone and N-(-hydroxyethyl)pyrrolidone; urea;
N,N-diethyl-m-toluamide; C.sub.2-C.sub.6 alkanediols; miscellaneous
solvents such as dimethyl formamide (DMF), N,N-dimethylacetamide
(DMA) and tetrahydrofurfuryl alcohol; and the 1-substituted
azacycloheptan-2-ones, particularly
1-n-dodecylcyclazacycloheptan-2-one (laurocapram; available under
the trademark Azone.RTM. from Whitby Research Incorporated,
Richmond, Va.).
[0816] Examples of solubilizers include, but are not limited to,
the following: hydrophilic ethers such as diethylene glycol
monoethyl ether (ethoxydiglycol, available commercially as
Transcutol.RTM.) and diethylene glycol monoethyl ether oleate
(available commercially as Softcutol.RTM.); polyethylene castor oil
derivatives such as polyoxy 35 castor oil, polyoxy 40 hydrogenated
castor oil, etc.; polyethylene glycol, particularly lower molecular
weight polyethylene glycols such as PEG 300 and PEG 400, and
polyethylene glycol derivatives such as PEG-8 caprylic/capric
glycerides (available commercially as Labrasol.TM.); alkyl methyl
sulfoxides such as DMSO; pyrrolidones such as 2-pyrrolidone and
N-methyl-2-pyrrolidone; and DMA. Many solubilizers can also act as
absorption enhancers. A single solubilizer may be incorporated into
the formulation, or a mixture of solubilizers may be incorporated
therein.
[0817] Suitable emulsifiers and co-emulsifiers include, without
limitation, those emulsifiers and co-emulsifiers described with
respect to microemulsion formulations. Emollients include, for
example, propylene glycol, glycerol, isopropyl myristate,
polypropylene glycol-2 (PPG-2) myristyl ether propionate, and the
like.
[0818] Other active agents may also be included in formulations,
e.g., other anti-inflammatory agents, analgesics, antimicrobial
agents, antifungal agents, antibiotics, vitamins, antioxidants, and
sunblock agents commonly found in sunscreen formulations including,
but not limited to, anthranilates, benzophenones (particularly
benzophenone-3), camphor derivatives, cinnamates (e.g., octyl
methoxycinnamate), dibenzoyl methanes (e.g., butyl methoxydibenzoyl
methane), p-aminobenzoic acid (PABA) and derivatives thereof, and
salicylates (e.g., octyl salicylate).
[0819] In certain topical formulations, the active agent is present
in an amount in the range of approximately 0.25 wt. % to 75 wt. %
of the formulation, preferably in the range of approximately 0.25
wt. % to 30 wt. % of the formulation, more preferably in the range
of approximately 0.5 wt. % to 15 wt. % of the formulation, and most
preferably in the range of approximately 1.0 wt. % to 10 wt. % of
the formulation.
[0820] Topical skin treatment compositions can be packaged in a
suitable container to suit its viscosity and intended use by the
consumer. For example, a lotion or cream can be packaged in a
bottle or a roll-ball applicator, or a propellant-driven aerosol
device or a container fitted with a pump suitable for finger
operation. When the composition is a cream, it can simply be stored
in a non-deformable bottle or squeeze container, such as a tube or
a lidded jar. The composition may also be included in capsules such
as those described in U.S. Pat. No. 5,063,507. Accordingly, also
provided are closed containers containing a cosmetically acceptable
composition as herein defined.
[0821] In an alternative embodiment, a pharmaceutical formulation
is provided for oral or parenteral administration, in which case
the formulation may comprises an activating compound-containing
microemulsion as described above, but may contain alternative
pharmaceutically acceptable carriers, vehicles, additives, etc.
particularly suited to oral or parenteral drug administration.
Alternatively, an activating compound-containing microemulsion may
be administered orally or parenterally substantially as described
above, without modification.
[0822] Cells, e.g., treated ex vivo with a compound described
herein, can be administered according to methods for administering
a graft to a subject, which may be accompanied, e.g., by
administration of an immunosuppressant drug, e.g., cyclosporin A.
For general principles in medicinal formulation, the reader is
referred to Cell Therapy: Stem Cell Transplantation, Gene Therapy,
and Cellular Immunotherapy, by G. Morstyn & W. Sheridan eds,
Cambridge University Press, 1996; and Hematopoictic Stem Cell
Therapy, E. D. Ball, J. Lister & P. Law, Churchill Livingstone,
2000.
[0823] Also provided herein are kits, e.g., kits for therapeutic
and/or diagnostic purposes. A kit may include one or more compounds
described herein, and optionally devices for contacting tissue or
cells with the compounds. Devices include needles, syringes,
stents, resuspension liquid, and other devices for introducing a
compound into a subject.
[0824] In any of the forgoing embodiments
1,5-bis(p-aminophenoxy)pentane may be specifically excluded.
[0825] In any of the forgoing embodiments 11-cis-retinol may be
specifically excluded.
[0826] In any of the forgoing embodiments 11-cis-retional palmitate
may be specifically excluded.
[0827] In any of the forgoing embodiments 13-cis-retinoic acid
(accutane) may be specifically excluded.
[0828] In any of the forgoing embodiments 2-bromopalmitic acid may
be specifically excluded.
[0829] In any of the forgoing embodiments 3-aminobenzoic acid ethyl
ester methane sulfonate may be specifically excluded.
[0830] In any of the forgoing embodiments acetaminophen may be
specifically excluded.
[0831] In any of the forgoing embodiments adamantylamine may be
specifically excluded.
[0832] In any of the forgoing embodiments all-trans-retinaldehyde
may be specifically excluded.
[0833] In any of the forgoing embodiments all-trans-retinoic acid
may be specifically excluded.
[0834] In any of the forgoing embodiments all-trans-retinol
(vitamin A) may be specifically excluded.
[0835] In any of the forgoing embodiments all-trans-retinyl
plamitate may be specifically excluded.
[0836] In any of the forgoing embodiments analine may be
specifically excluded.
[0837] In any of the forgoing embodiments cyclohexylamine may be
specifically excluded.
[0838] In any of the forgoing embodiments dapson may be
specifically excluded.
[0839] In any of the forgoing embodiments diaminophenoxypentane may
be specifically excluded.
[0840] In any of the forgoing embodiments ethyl m-aminobenzoate may
be specifically excluded.
[0841] In any of the forgoing embodiments m-aminobenzoic acid may
be specifically excluded.
[0842] In any of the forgoing embodiments m-phenetidine may be
specifically excluded.
[0843] In any of the forgoing embodiments
N-(4-hydroxyphenyl)retinamide (fenretinide) may be specifically
excluded.
[0844] In any of the forgoing embodiments N,N-dimethylaniline may
be specifically excluded.
[0845] In any of the forgoing embodiments
N,N-dimethyl-p-phenetidine may be specifically excluded.
[0846] In any of the forgoing embodiments N-methylaniline may be
specifically excluded.
[0847] In any of the forgoing embodiments N-methyl-p-phenetidine
may be specifically excluded.
[0848] In any of the forgoing embodiments o-phenetidine may be
specifically excluded.
[0849] In any of the forgoing embodiments p-(n-hexyloxy)aniline may
be specifically excluded.
[0850] In any of the forgoing embodiments p-(n-hexyloxy)benzamide
may be specifically excluded.
[0851] In any of the forgoing embodiments p-(n-hexyloxy)benzoic
acid hydrazide may be specifically excluded.
[0852] In any of the forgoing embodiments p-anisidine may be
specifically excluded.
[0853] In any of the forgoing embodiments p-ethylanaline may be
specifically excluded.
[0854] In any of the forgoing embodiments p-ethyoxybenzylamine may
be specifically excluded.
[0855] In any of the forgoing embodiments p-ethyoxyphenol may be
specifically excluded.
[0856] In any of the forgoing embodiments phenetidine may be
specifically excluded.
[0857] In any of the forgoing embodiments piperidine may be
specifically excluded.
[0858] In any of the forgoing embodiments p-n-boutoxyaniline may be
specifically excluded.
[0859] In any of the forgoing embodiments p-n-butylaniline may be
specifically excluded.
[0860] In any of the forgoing embodiments p-n-dodecylaniline may be
specifically excluded.
[0861] In any of the forgoing embodiments p-nitroaniline may be
specifically excluded.
[0862] In any of the forgoing embodiments sulfabenzamide may be
specifically excluded.
[0863] In any of the forgoing embodiments sulfamoxaole may be
specifically excluded.
[0864] In any of the forgoing embodiments sulfanilamide may be
specifically excluded.
[0865] In any of the forgoing embodiments tricaine may be
specifically excluded.
[0866] In addition, any compound cited in the references
incorporated herein may also be specifically excluded from any of
the forgoing embodiments.
[0867] 4. Methods
[0868] Disclosed herein are methods for treating or preventing an
ophtalmologic disorder. An exemplary method comprises administering
to a subject, e.g. a subject in need thereof, a therapeutically
effective amount of a composition, e.g., a pharmaceutical
composition, described herein. A subject in need thereof may be a
subject who knows that he has or is likely to develop an
opthalmologic disorder.
[0869] As discussed above, a disclosed composition may be
administered to a subject in order to treat or prevent macular
degeneration. Other diseases, disorders, or conditions
characterized by the accumulation of retinotoxic compounds, e.g.,
lipofuscin, in the RPE may be similarly treated (e.g.,
lipofuscin-based retinopathies).
[0870] The methods described herein may be used for the treatment
or prevention of any form of retinal or macular degeneration
associated with lipofuscin accumulation, such as hereditary or
degenerative diseases of the macula as e.g. age-related macular
degeneration (AMD). There are two forms of age-related macular
degeneration, dry (atrophic) and wet (neovascular or exudative)
macular degeneration.
[0871] As discussed above, macular degeneration (also referred to
as retinal degeneration) is a disease of the eye that involves
deterioration of the macula, the central portion of the retina.
Approximately 85% to 90% of the cases of macular degeneration are
the "dry" (atrophic or non-neovascular) type.
[0872] In "dry" macular degeneration, the deterioration of the
retina is associated with the formation of small yellow deposits,
known as drusen, under the macula. This phenomena leads to a
thinning and drying out of the macula. The location and amount of
thinning in the retinal caused by the drusen directly correlates to
the amount of central vision loss. Degeneration of the pigmented
layer of the retina and photoreceptors overlying drusen become
atrophic and cause a slow of central vision. This often occurs over
a decade or more. Vision loss can occur very rapidly in subjects
having strong geographic atrophy. Such subjects may be treated as
described herein.
[0873] Most people who lose vision from age related macular
degeneration have "wet" macular degeneration. In "wet"
(neovascular) macular degeneration, abnormal blood vessels from the
choroidal layer of the eye, known as subretinal neovascularization
grow under the retina and macula. These blood vessels tend to
proliferate with fibrous tissue, and bleed and leak fluid under the
macula, causing the macula to bulge or move and distort the central
vision. Acute vision loss occurs as transudate or hemorrhage
accumulates in and beneath the retina. Permanent vision loss occurs
as the outer retina becomes atrophic or replaced by fibrous
tissues.
[0874] Stargardt's disease (STGD) is a recessive form of macular
degeneration with an onset during childhood. STGD is characterized
clinically by progressive loss of central vision and progressive
atrophy of the retinal pigment epithelium (RPE) overlying the
macula. Mutations in the human ABCR gene for RmP are responsible
for STGD. Early in the disease course, patients show delayed dark
adaptation but otherwise normal rod function. Histologically, STGD
is associated with deposition of lipofuscin pigment granules in RPE
cells, presumably arising from impaired digestion after
phagocytosis of shed distal outer-segments. Degeneration of the RPE
occurs subsequently, with photoreceptor degeneration appearing late
in the disease. This pathological picture has lead to the
conclusion that STGD is primarily a defect of the RPE. However, the
pattern of early RPE degeneration and preservation of
photoreceptors must be reconciled with the observation that RmP is
present exclusively in outer segments and not expressed in RPE
cells. Some AMDs are caused by mutations in a gene, such as the
genes ABCA4, ELOVL4, PROML1, VMD2, Peripherin/RDS, EFEMP1, TIMP3,
and XLRS1. One mutation of the ABC4A gene is G1961E. Macular
dystrophies also include the following diseases: Stargardt
disease/fundus flavimaculatus (OMIM 248200), which is an autosomal
recessive disease characterized by a mutation at chromosome locus
1p21-p22 (STGD1); Stargardt-like macular dystrophy (OMIM 600110),
which is an autosomal dominant disease characterized by a mutation
at chromosome locus 6q14 (STGD3); Stargardt-like macular dystrophy
(OMIM 603786), which is an autosomal dominant disease characterized
by a mutation at chromosome locus 4p (STGD4); autosomal dominant
"bull's eye" macular dystrophy, which is an autosomal dominant
disease characterized by a mutation at chromosome locus 4p (MCDR2);
Bestmacular dystrophy (OMIM 153700), which is an autosomal dominant
disease characterized by a mutation at chromosome locus 11q13;
adult vitelliform dystrophy (OMIM 179605), which is an autosomal
dominant disease characterized by a mutation at chromosome locus
6p21.2-cen; pattern dystrophy (OMIM 169150), which is an autosomal
dominant disease characterized by a mutation at chromosome locus
6p21.2-cen; Doyne honeycomb retinal dystrophy (OMIM 126600), which
is an autosomal dominant disease characterized by a mutation at
chromosome locus 2p16; North Carolina macular dystrophy (OMIM
136550), which is an autosomal dominant disease characterized by a
mutation at chromosome locus 6q14-q16.2 (MCDR1); autosomal dominant
macular dystrophy resembling MCDR1, which is an autosomal dominant
disease characterized by a mutation at chromosome locus
5p15.33-p13.1 (MCDR3); North Carolina-like macular dystrophy
associated with deafness, which is an autosomal dominant disease
characterized by a mutation at chromosome locus 14p (MCDR4);
progressive biforcal chorioretinal atrophy (OMIM 600790), which is
an autosomal dominant disease characterized by a mutation at
chromosome locus 6q14-q16.2; Sorby's fundus dystrophy (OMIM
136900), which is an autosomal dominant disease characterized by a
mutation at chromosome locus 22q12.1-q13.2; central areolar
choroidal dystrophy (OMIM 215500), which is an autosomal dominant
disease characterized by a mutation at chromosome locus
6p21.2-cen17p13; dominant cystoid macular dystrophy (OMIM 153880),
which is an autosomal dominant disease characterized by a mutation
at chromosome locus 7p15-p21; and juvenile retinoschisis (OMIM
312700), which is an X-linked disease characterized by a mutation
at chromosome locus Xp22.2 (Michaelides et al. (2003) J. Med.
Genet. 40:641). The methods described herein may be used to treat
or prevent any of these genetically inhereted macular dystrophies,
provided that they are associated with abnormal lipofuscin
accumulation similar to AMD and Stargardt disease.
[0875] Other diseases that may be treated or prevented include
cone-rod dystrophy, certain types of retinitis pigmentosa, and
fundus flavimaculatus.
[0876] In one embodiment, a drug is administered to a subject that
short-circuits the visual cycle at a step of the visual cycle that
occurs outside a disc of a rod photoreceptor cell. For example, as
shown in FIG. 3, the drug may react with 11-cis-retinal in the RPE
and shunt it to all-trans-retinal while it remains in the RPE. More
specifically, the therapeutic may react with 11-cis-retinal to form
an intermediate that isomerizes to the all-trans configuration. The
all-trans intermediate may then release the therapeutic to form
all-trans-retinal. The all-trans-retinal could then be re-processed
through the remainder of the visual cycle as normal in the RPE.
Thus, the visual cycle would be reduced to a futile cycle, in which
all-trans-retinal has little or no opportunity to accumulate in the
disc.
[0877] In one embodiment, a subject may be diagnosed as having
macular degeneration, and then a disclosed drug or combination
therapy may be 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). In yet another embodiment, a subject may be
diagnosed as having Stargardt's disease, a familial form of macular
degeneration. In some embodiments, a drug may be administered
prophylactically. In some embodiments, a subject may be diagnosed
as having the disease before retinal damage is apparent. For
example, a subject may be found to carry a gene mutation for abcr,
elovl4, and/or another gene, and thus be diagnosed as having
Stargardt's disease before any opthalmologic signs are manifest, or
a subject may be found to have early macular changes indicative of
macular degeneration before the subject is aware of any effect on
vision. In some embodiments, a human subject may know that he or
she is in need of the macular generation treatment or
prevention.
[0878] Doctors can usually diagnose macular degeneration by
examining the eyes with an opthalmoscope or a slit lamp. Sometimes
fluorescein angiography--a procedure in which a doctor injects dye
into a vein and photographs the retina--is used to determine the
diagnosis. Lipofuscin accumulation may be detected by
autofluorescence imaging optionally with confocal scanning laser
opthalmoscope.
[0879] 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,
angiography, fluorescein angiography, and/or biopsy. Monitoring can
be performed at a variety of times. For example, a subject may be
monitored after a drug is administered. The monitoring can occur
one day, one week, two weeks, one month, two months, six months,
one year, two years, and/or five years after the first
administration of a drug. A subject can be repeatedly monitored. In
some embodiments, the dose of a drug may be altered in response to
monitoring.
[0880] In some embodiments, the disclosed methods may be combined
with other methods for treating or preventing macular degeneration,
such as photodynamic therapy. Other methods are further described
herein.
[0881] In some embodiments, a drug for treating or preventing
macular degeneration may be administered chronically. The drug may
be administered daily, more than once daily, twice a week, three
times a week, weekly, biweekly, monthly, bimonthly, semiannually,
annually, and/or biannually.
[0882] The therapeutics may be administered by a wide variety
routes, described above. In some embodiments, a drug may be
administered orally, in the form of a tablet, a capsule, a liquid,
a paste, and/or a powder. In some embodiments, a drug may be
administered locally, as by intraocular injection. In some
embodiments, a drug may be administered systemically in a caged,
masked, or otherwise inactive form and activated in the eye (such
as by photodynamic therapy). In some embodiments, a drug may be
administered in a depo form, so sustained release of the drug is
provided over a period of time, such as hours, days, weeks, and/or
months.
[0883] The therapeutic agents are used in amounts that are
therapeutically effective, which varies widely depending largely on
the particular agent being used. The amount of agent incorporated
into the composition also depends upon the desired release profile,
the concentration of the agent required for a biological effect,
and the length of time that the biologically active substance has
to be released for treatment. In certain embodiments, the
biologically active substance may be blended with a compound matrix
at different loading levels, in one embodiment at room temperature
and without the need for an organic solvent. In other embodiments,
the compositions may be formulated as microspheres. In some
embodiments, the drug may be formulated for sustained release.
[0884] It is noted that disruption of the visual cycle to prevent
accumulation of A.sub.2E may impair a subject's night (low-light)
vision and might cause night-blindness. Indeed, some of the
therapeutics noted herein as appropriate for preventing A.sub.2E
accumulation have been used sparingly in humans or withheld from
use entirely because of their propensity to cause night-blindness.
However, with the recognition that this very cause of night
blindness might be turned to the therapeutic and/or preventative
treatment of macular degeneration, it is likely that patients in
need of such treatment would readily accept some night-blindness in
return for sparing of normal vision. This is because the visual
cycle described above operates in rod photoreceptors, which operate
only at low levels of illumination and do not operate during the
day. Therefore, macular function would be little affected by
decreases in visual cycle function, while there might be some
effect on low light vision at night. At least some patients, and
probably most, might readily sacrifice a decrement in night vision
for a lessening of the probability that they would eventually lose
their cone day vision.
[0885] 5. Screening Methods
[0886] Suitable drugs may be identified by a variety of screening
methods. For example, a candidate drug may be administered to a
subject that has or is at risk for having macular degeneration,
e.g., an animal that is an animal model for macular degeneration,
and the accumulation of a retinotoxic compound, such as A.sub.2E,
can be measured. A drug that results in reduced accumulation of a
retinotoxic compound compared to a control (absence of the drug)
would thus be identified as a suitable drug. Alternatively,
photoreceptor disks may be analyzed for the presence of
all-trans-retinal, N-retinylidene-PE, and/or A.sub.2E. Animal
models that have rapid development of macular degeneration are of
considerable interest because naturally-occurring macular
degeneration typically takes years to develop. A number of animal
models are accepted models for macular degeneration. For example,
the abcr -/- knockout mouse has been described as a model for
macular degeneration and/or lipofuscin accumulation, as has been
the elovl4-/- knockout mouse. In addition, knockout mice deficient
in monocyte chemoattractant protein-1 (Ccl-2; also known as MCP-1)
or it cognate receptor, C--C chemokine receptor-2 (Ccr-2), have
also been described as accelerated models for macular
degeneration.
[0887] In addition, in vitro models of the visual system may
facilitate screening studies for drugs that inhibit or short
circuit the visual cycle. In vitro models can be created by placing
selected intermediates in solution with appropriate enzymes and
other necessary cofactors. Alternatively, an in vitro RPE culture
system may be employed. For example, LRAT inhibition can be tested
by adding a candidate drug to a solution containing LRAT and a
substrate for LRAT, and measuring accumulation of an expected
product. Analogous systems are envisioned for the other potential
inhibition targets described herein.
[0888] Agents, such as small molecules, that inhibits one of the
enzymes of the visual cycle or shortcircuits the visual cycle may
also be identified. For example, agents that bind to one of these
enzymes may be identified. Screening assays may comprise contacting
an enzyme with a test agent and determining whether the agent binds
to the enzyme. An enzyme of biologically active fragment thereof
may be used. The enzyme or fragment may be labeled, such as with a
fluorophore. Screening assays may also comprise contacting an
enzyme with its substrate in the presence of a test agent and
determine whether the test agent prevents binding of the enzyme to
the substrate. These assays may be adapted to highthroughput
screening assays.
[0889] Agents, such as small molecules, that inhibit vitamin A
delivery to the eye may be identified in screening methods. A
method may comprise contacting an RBP with a test agent in the
presence of vitamin A and measuring the amount of vitamin A bound
to RBP in the presence relative to the absence of the test agent.
Another method may comprise contacting an RBP with a test agent in
an vitro eye model, adding vitamin A, and determining the amount of
vitamin A that is imported into the eye model in the presence
relative to the absence of the test agent. Other screening assays
may comprise contacting an RBP and an RBP receptor optionally in
the presence of vitamin A and determining the amount of RBP bound
to the RBP receptor in the presence relative to the absence of the
test agent.
EXAMPLES
[0890] The present description is further illustrated by the
following examples, which should not be construed as limiting in
any way. The contents of all cited references (including literature
references, issued patents, published patent applications as cited
throughout this application) are hereby expressly incorporated by
reference.
Example 1
TDT and TDH Bind to mRPE65 with High Affinities
[0891] Materials: Frozen bovine eye-cups devoid of retinas were
purchased from W. L. Lawson Co., Lincoln, Nebr.
Ethylenediaminetetraacetic acid (EDTA), phenyl-Sepharose CL-4B, and
Trizma.RTM. base, trans-trans-farnesol, pyridinium chlorochromate,
Dess-Martin reagent, decyl magnesium bromide, hexadecyl amine,
dimethylsulfoxide were from Sigma-Aldrich. Dithiothreitol (DTT) was
from ICN Biomedicals Inc. Anagrade.TM. CHAPS was from Anatrace.
HPLC grade solvents were from Sigma-Aldrich Chemicals. Anti RPE65
(NFITKVNPETLETIK) antibody was obtained from Genmed Inc. Broad
spectrum EDTA-free protease inhibitor cocktail was obtained from
Roche Biosciences. The precast gels (4-20%) for SDS-PAGE, BenchMark
prestained molecular weight marker were from Invitrogen. DEAE
Sepharose was from Amersham Biosciences. All reagents were
analytical grade unless specified otherwise.
[0892] Methods
[0893] Animal Studies: Protocols were approved by the Standing
Committee on Animal Care of Harvard Medical School, the
Institutional Animal Care and Use Committee of Columbia University
and complied with guidelines set forth by The Association for
Research in Vision and Opthalmology. 7 week old male Balb/c albino
mice and 7 week old male Sprague-Dawley rats were from Charles
River Breeding Laboratories and were housed in 12:12 h light:dark
cycle. 8-10 week old Abcr null mutant mice (129/SV X C57BL/6J) were
generated as formerly described (12,20) and Abcr.sup.-/- and
Abcr.sup.+/+ mice were raised under 12-hour on-off cyclic lighting
with an in-cage illuminance of 30-50 lux. In Abcr.sup.-/- and
Abcr.sup.+/+ mice, Rpe65 was 9 sequenced as reported previously
[20].
[0894] Purification of mRPE65: mRPE65 was extracted and purified
from the bovine eye cups using a procedure described earlier [29].
Protein purity was established by silver staining and Western
blotting (1:4000 primary antibody-1 h at room temperature and
1:4000 secondary antibody 0.5 h at room temperature). Buffers were
changed by dialysis in the request buffer overnight in a
Slide-a-lyser.TM. cassette from Pierce (10 KDa MWCO). RPE65
solutions were concentrated with an Amicon Ultra.TM. centrifugal
filtration device (30 KDa-cutoff) from Millipore Corp. The final
protein solution contained 100 mM phosphate buffered saline (150 mM
NaCl) pH 7.4 and 1% CHAPSO. The protein concentration was measured
by a modified Lowry method [30] using the Bio-Rad DC protein assay
protocol.
[0895] Syntheses:
[0896] 13,17,21-Trimethyl-docosa-12,16,20-trien-11-one (TDT: A
solution of trans, trans-farnesal (200 mg, 0.9 mmol) in ether (2
mL) was added to a solution of decyl magnesium bromide (1 M
solution in ether, 1.5 mL) at 0.degree. C. and stirred for 15 min.
The reaction mixture was then warmed to room temperature and
quenched with aqueous saturated NH.sub.4Cl (1 mL). H.sub.2O (2 mL)
was added and the reaction mixture was extracted with hexane
(3.times.5 mL). The combined extracts were collected, washed with
brine, dried with magnesium sulfate and evaporated under reduced
pressure. The residue was chromatographed (SiO.sub.2, EtOAc-light
petroleum, 10:90) to give the alcohol (319 mg, 92%); R.sub.f
(EtOAc-light petroleum, 2:8) 0.56. Dess-Martin periodinate (419 mg,
0.99 mmol) was added to a solution of the above alcohol (319 mg,
0.83 mmol) in CH.sub.2Cl.sub.2 (1.5 mL) at room temperature and
stirred for 10 min. The reaction mixture was then treated with
sodium thiosulfate-sodium bicarbonate solution (1:1 v/v of 10%
sodium thiosulfate and aqueous saturated NaHCO.sub.3, 3 mL) and
stirring continued for another 10 min. H.sub.2O (2 mL) was added,
the reaction mixture was extracted with hexane (3.times.5 mL),
washed with brine, dried with Mg.sub.2SO4 and the combined extracts
were evaporated under reduced pressure. The residue was
chromatographed (SiO.sub.2, EtOAc-light petroleum, 1:99) to give
the ketone (TDT) (283 mg, 89%); R.sub.f(EtOAc-light petroleum, 2:8)
0.8; .delta..sub.H (200 MHz; CDCl.sub.3) 6.04 (s, 1H), 5.19-5.01
(m, 2H), 2.44-2.30 (m, 2H), 2.20-1.85 (m, 8H), 1.71 (s, 3H), 1.59
(s, 6H), 1.55 (s, 3H) and 1.38-1.17 (m, 21H); m/z (ESI) (Found:
M+Na 383.3277, C.sub.25H.sub.44O requires M+Na 383.3284).
[0897] 3,7,11-Trimethyl-dodeca-2,6,10-trienoic acid hexadecylamide
(TDH): NaCN (31 mg) and MnO.sub.2 (590 mg) were added to a stirring
solution of trans-trans-farnesol (100 mg, 0.45 mmol) in hexane (3
mL) at room temperature, followed by hexadecyl amine (545 mg, 2.2
mmol) and stirring continued for 1 h. An additional portion of
MnO.sub.2 (590 mg) was added and the mixture left for overnight at
room temperature with stirring. The mixture was then filtered
through a pad of silica and celite and washed with hexane several
times. The combined extracts were evaporated and the residue was
chromatographed (SiO.sub.2, EtOAc-light petroleum, 3:97) to give
3,7,11-Trimethyl-dodeca-2,6,10-trienoic acid hexadecylamide (145
mg, 70%); R.sub.f(EtOAc-light petroleum, 2:8) 0.52; .delta..sub.H
(200 MHz; CDCl.sub.3) 8.18 (d, J=9 Hz, 1H), 6.0 (d, J=9.6 Hz, 1H),
5.2-5.0 (m, 2H), 3.42 (t, J=6.8 Hz, 2H), 2.23-1.91 (m, 8H), 1.67
(s, 3H), 1.59 (s, 9H), 1.39-1.20 (m, 31H); m/z (ESI) (Found: M+Na
482.4334, C.sub.31H.sub.57ON requires M+Na 482.2332).
[0898] Fluorescence binding assays: RPE65 in PBS, 1% CHAPS, pH 7.4
was used in the fluorometric titration studies. All titrations were
performed at 25.degree. C. The samples in PBS buffer were excited
at 280 nm and the fluorescence was scanned from 300 to 500 nm.
Fluorescence measurements, using 450 .mu.L quartz cuvettes with a
0.5 cm path length, were made at 25.degree. C. on a Jobin Yvon
Instruments, Fluoromax 2 employing the right-angle detection
method.
[0899] The fluorescence of the protein solution was measured after
equilibrating it at 25.degree. C. for 10 min. The sample was then
titrated with a solution of retinoid dissolved in DMSO in the
absence of any overhead light and the solution was mixed thoroughly
before fluorescence measurement. In each titration, to a 350 .mu.L
solution of the protein an equivalent amount of ligand, typically
0.3 .mu.L. was added and thoroughly mixed before allowing it to
equilibrate for 10 min prior to recording the fluorescence
intensity. The addition of DMSO (0.1% per addition) did not have
any effect on the fluorescence intensity. The binding constant
(K.sub.D) was calculated from the fluorescence intensity as
described before (17,19).
[0900] Drug treatment and retinoid extraction: The drugs were
injected i.p in DMSO as carrier. Controls received DMSO alone. The
volume of the solution was 50 .mu.L for mice and 180 .mu.L for
rats. After the injections were given the animals were housed in
dark for 2 h and then bleached for 2 h. Then the animals were dark
adapted (5 min for mice and 30 min for rats) before being
sacrificed and eyes were enucleated. In the experiments performed
male Balb/c mice and male Sprague Dawley rats were used.
[0901] Eyes were placed in glass-glass homogenizer in 0.8 mL of 1 M
hydroxylamine/0.1 M MOPS [3-(N-morpholino) propanesulfonic acid],
pH 6.5 and 0.2% SDS and homogenized [31]. Ethanol (0.6 mL) was
added and the homogenates were incubated for 30 min at room
temperature to allow formation of the 11-cis-retinal oximes [31].
The retinoids were extracted with dichloromethane (3.times.0.4 mL).
The combined extracts were dried with magnesium sulfate, evaporated
under the flow of argon and subjected to HPLC analysis. The normal
phase HPLC column was YMC-PVA SIL NP, 250.times.4.6 mm and the
mobile phase was hexane-dioxane (93:7 v/v) with a flow rate of 1.5
mL/min. Absorbance was monitored at 325 nm, and peaks were
identified by comparing with standards. In the HPLC profiles shown,
retinyl esters and 11-cis-retinal syn-oxime were measured.
Regeneration time was 5 min with mice and 30 min with rats. The
rates of regeneration and the effects of the drugs appeared
indistinguishable between the Balbc/mice and the pigmented wt
(129/SV X C57BL/6J) mice (on the same genetic background as the
Abcr knockout mice).
[0902] Electroretinogram Determinations (ERG): Mice were
dark-adapted overnight before all ERG experiments. To determine the
acute effect of compounds under study, mice were given a single
i.p. injection of a compound at 50 mg/kg in 25 uL DMSO under dim
red light and kept in darkness for an additional 1 h before being
exposed to the bleaching light prior to ERG recordings. Control
("untreated") animals were injected with 25 uL of DMSO. Mice were
anaesthetized with ketamine (80 mg/kg) and xylazine (5-10 mg/kg)
and pupils were dilated with 1% phenylephrine and 1%
cyclopentolate, followed by an exposure to 5000 lux bleaching light
for 2 min. The ERG was recorded from the cornea with a cotton wick
saline electrodes for about 50 min immediately after bleaching.
Subcutaneous 30 gauge needles on the forehead and trunk were used
as reference and ground electrodes, respectively. The animals
rested on a heater maintaining the body temperature at 37.degree.
C. The light stimulus was obtained from a ganzfeld stimulator
having a stroboscope (PS33 Grass Instruments Inc., West Warwick,
R.I.) removed from its housing and recessed above and behind the
head of the mouse. The flash was diffused to cover the ganzfeld
homogeneously. Maximum flash intensity was measured with a
calibrated light meter (J16 Tektronics Instruments, Beaverton,
Oreg.). Responses were averaged by a Macintosh computer-controlled
data acquisition system (PowerLab, AD Instruments, Mountain View,
Calif.) at a frequency of 0.1 Hz.
[0903] The same animals were subjected to ERG experiments according
to exactly the same protocol 3 days later, except no (repeated)
injection of compounds was performed.
[0904] Tissue extraction and HPLC analysis: Posterior eye cups were
pooled and homogenized in PBS using a tissue grinder. An equal
volume of a mixture of chloroform/methanol (2:1) was added and the
sample was extracted three times. To remove insoluble material,
extracts were filtered through cotton and passed through a reversed
phase (C18 Sep-Pak, Millipore) cartridge with 0.1% TFA in methanol.
After removing solvent by evaporation under gas, the extract was
dissolved in methanol containing 0.1% TFA, for HPLC analysis. For
quantification of A.sub.2E, a Waters Alliance 2695 HPLC was
employed with a Atlantis.RTM. dC18 column (Waters, 4.6.times.150
mm, 3 .mu.m) and the following gradient of acetonitrile in water
(containing 0.1% trifluoroacetic acid): 90-100% (0-10 min), 100%
acetonitrile (10-20 min), and a flow rate of 0.8 mL/min with
monitoring at 430 nm. The injection volume was 10 .mu.L. Extraction
and injection for HPLC were performed under dim red light. Levels
of A.sub.2E and iso-A.sub.2E were determined by reference to an
external standard of HPLC-purified A.sub.2E/iso-A.sub.2E. Since
A.sub.2E and iso-A.sub.2E reach photoequilibrium in vivo [4], use
of the term A.sub.2E will refer to both isomers, unless stated
otherwise.
[0905] Results
[0906] Design and In Vitro Activities of Specific mRPE65
Antagonists: In previous quantitative fluorescence studies we
showed that mRPE65 saturably binds all-trans-retinyl palmitate
(tRP) with a K.sub.D=47 nM [17,19]. Further structure-activity
studies on ligand binding to mRPE65 reveals that amide and ketone
equivalents of tRP bound approximately as well as tRP itself
[unpublished data]. Moreover, isoprenoids, such as C15 farnesyl,
can substitute for the all-trans-retinyl moiety [unpublished data].
Based on these observations we prepared the trans,
trans-farnesylated ketone (TDT) and amide (TDH) shown in FIGS.
4A-B. TDT and TDH specifically bind to purified bovine mRPE65 as
shown by fluorescence titration of mRPE65 with TDT and TDH reported
in FIGS. 4A-B. The excitation wavelength was at 280 nm and the
emission was observed through 0.5 cm layer of solution. The
titration solution consisted of 0.952 .mu.M of mRPE65 in 100 mM
phosphate buffered saline (150 mM NaCl) pH 7.4 and 1% CHAPS. Panel
(a) of FIG. 4A shows the emission spectra of mRPE65 when binding to
TDT. Panel (b) shows the change in the fluorescence intensity at
338 nm with increasing concentrations of TDT or TDH. Panel (c)
shows the linear square fit plots of the equation P.sub.0.alpha. vs
R.sub.0.alpha./(1-.alpha.), for the titration of mRPE65 vs. TDT or
TDH [17,19]. TDT binds with K.sub.D=58.+-.5 nM while TDH binds with
K.sub.D=96.+-.14 nM. In these experiments, we made use of the fact
that the specific binding of these analogs to mRPE65 quenches
protein fluorescence.
[0907] In Vivo Studies with Analogs TDT and TDH
[0908] Acute effects: In order to determine whether TDT and TDH
have an effect on the visual cycle in vivo, the overnight
dark-adapted Abca4.sup.+/+ (Rpe65 450Leu, pigmented, 129/SV X
C57BL/6J) mice received single (i.p.) injections of the two
compounds at 50 mg/kg. For comparison, mice were also injected with
13-cis-retinoic acid (13-RA; Accutane) at the same concentration.
One hour after treatment, the mice were subjected to
photo-bleaching (5000 lux for 2 min to bleach .about.90% of
rhodopsin) and ERGs were recorded.
[0909] FIG. 5A shows the effects of TDT and TDH on the ERG b-wave
amplitudes in animals 1 h after treatment with 50 mg/kg dose of
three compounds (13-RA, TDT and TDH) on rod b-wave amplitude
recovery after photo-bleaching. Results are averaged from three
mice in each group with SD bars shown. Both isoprenoids delayed the
recovery of dark-adapted visual responses to an extent similar to
that of 13-RA as judged from dark-adapted ERG b-wave amplitudes
recorded using dim light flashes delivered immediately before and
at regular intervals after photo-bleaching. A substantial effect on
rod b-wave recovery induced by TDT and TDH was still present 3 days
after treatment while no sustained effect of 13-RA was detected
(FIG. 5B).
[0910] To establish whether recovery of the dark-adapted rod b-wave
was retarded because of an effect on 11-cis-retinal synthesis we
studied the effects of TDT on 11-cis-retinal regeneration in rats
and mice. The 11-cis-retinal that is regenerated is essentially all
bound to rhodopsin in rodents, so that measuring its level is
equivalent to measuring the rhodopsin content [27]. Initial
experiments were performed on Sprague Dawley rats because similar
experiments using 13-RA were previously carried out using these
animals so that ready comparisons of potency and effectiveness can
be made [24]. In these experiments, 13-RA was shown to exhibit
profound effects on visual cycle function by interfering with
11-cis-retinal regeneration after a bleaching [24]. Accordingly, in
the current experiments, the rats were given single injections
(i.p.) of TDT (TDH) (50 mg/kg in DMSO), 13-RA (50 mg/kg in DMSO),
and DMSO alone. After injecting the analogs, the rats were dark
adapted for 2 h, and then exposed to light that led to <10% of
dark adapted 11-cis-retinal in these animals, compared to dark
adapted controls (data not shown). After allowing the bleached rats
to dark-adapt again for 30 min, the animals were sacrificed, and
the regenerated 11-cis-retinal was determined as indicated in
Methods. In these experiments, the amount of resynthesized
11-cis-retinal, the chromophore of rhodopsin, is measured by HPLC
and compared to the amounts of all-trans-retinyl ester
precursor.
[0911] As shown in FIGS. 6A-C, both 13-RA (FIG. 6A) and TDT (FIG.
6B) achieved substantial (4-5-fold) inhibition of 11-cis-retinal
regeneration, while the inhibitory effect of TDH (FIG. 6C) is less
pronounced than with TDT. These figures show results of HPLC
analysis of extracted retinoids from drug treated rats. FIG. 6A
shows HPLC data from 13-RA treated Sprague Dawley rats. In FIG. 6A,
the relative amounts of 11-cis-retinal-syn-oxime and
all-trans-(cis)-retinyl esters are shown for the control (--13-RA)
(top) and drug treated animals (bottom). In FIG. 6B, the
corresponding data are shown for the TDT treated rats (top-control,
bottom TDT treated). In this case, the ester pool in the drug
treated rat is largely, if not exclusively, all-trans. FIG. 6C
shows corresponding data for TDH treated rats (top-control, bottom
TDH treated). These results are consistent with the observed lower
potency of TDH as an mRPE65 antagonist compared to TDT. Upon
repetition of the experiments two further times the average
inhibition values are as follows: 13-RA (78.+-.2%), TDT (79.+-.4%),
and TDH (55.+-.2%). These percent inhibition values are generated
by comparing the integrated areas under the retinyl ester and
11-cis-retinal-syn-oxime peaks. (Materials and Methods).
[0912] It is significant that the all-trans-retinyl ester pool
increases at the expense of the 11-cis-retinal pool in the presence
of TDT and TDH (FIGS. 6A-C). The concomitant increase in the
all-trans-retinyl ester pool is expected of an antagonist of
mRPE65. The magnitude of inhibition by 13-RA is approximately the
same as reported [25]. In the case of 13-RA inhibition, the ester
pool is a mixture of the 11-cis and all-trans isomers because of
the inhibition of 11-cis-retinol-dehydrogenase [22,25]. In the
experiments described here with the isoprenoid antagonists, only
all-trans-retinyl esters are detectable [data not shown].
[0913] Similar experiments with inhibitors were also performed in
Balb/c mice. Here again inhibition is observed, but the effects are
less pronounced than in rats, both with 13-RA and the isoprenoid
antagonists. The inhibition values are as follows: 13-RA
(33.+-.4%), TDT (35.+-.2%), and TDH (24.+-.6%). It is noteworthy
that in rats the 11-cis chromophore is regenerated considerably
more slowly than in mice [27]. In mice, as in rats, the isoprenoid
mRPE65 antagonists and 13-RA proved to be approximately equi-potent
with respect to the inhibition of 11-cis-retinal regeneration.
[0914] b) Effect of the long-term (chronic) treatment of
ABAC4.sup.-/- mice with TDT and TDH on A.sub.2E accumulation: The
RPE65 antagonists TDT and TDH were further tested for their
abilities to reduce the accumulation of the RPE lipofuscin
fluorophores A.sub.2E and iso-A.sub.2E. Beginning at 2 months of
age, Abca4.sup.-/- mice (on the same genetic background as the
Abca4.sup.+/+ animals) were given i.p. injections of the two
compounds at 50 mg/kg twice a week and A.sub.2E and iso-A.sub.2E
levels were determined by quantitative HPLC after an additional 2
months.
[0915] As shown in FIGS. 7A-D, both compounds, but especially TDT,
were highly efficient in lowering A.sub.2E accumulation. FIGS. 7A-D
show quantitation of A.sub.2E and iso-A.sub.2E in eye cups of Abcr
-/- mice. (A-C) Typical chromatograms obtained by reverse-phase
HPLC with monitoring at 430 nm illustrate the detection of A.sub.2E
and iso-A.sub.2E and a reduction in peak intensity with TDT and TDH
treatment relative to vehicle-treated controls. FIG. 7D:
A.sub.2E/iso-A.sub.2E quantitation from integrated peak areas
normalized to external standards. Values expressed as picomoles per
eye and are based on single samples obtained by pooling 4 eyes.
[0916] Specifically, the levels of A.sub.2E in eyecups of mice
treated with TDT were 85% lower than in vehicle-treated (DMSO)
Abca4.sup.-/- animals. This is an under estimate of the extent of
A.sub.2E reduction because drug treatment did not commence until
two months of age, and the data is not corrected for A.sub.2E
accumulation in the knockout mice up to this point. These results
demonstrate that the mRPE65 antagonists TDT and TDH are effective
in vivo and slow the rate of A.sub.2E accumulation by limiting
visual cycle function.
[0917] Discussion
[0918] RPE65 is of central importance in the operation of the
visual cycle and been shown to be necessary for rhodopsin
regeneration [15]. The retinoic acids specifically bind to mRPE65
and can block isomerization in RPE membranes [22]. The fact that
13-RA limits the visual cycle in rats in vivo [24] suggests the
possibility that mRPE65 might be a viable target for interfering
with toxic lipofuscin formation. In rats, the effects of 13-RA on
visual function are pronounced. There is an approximately 4-fold
inhibition measured for 11-cis-retinal regeneration after bleaching
[24]. This inhibition is translated into a diminution in the
accumulation of the lipofuscins in the A.sub.2E series in the
Abca4.sup.-/- knockout mouse model [25]. However, since the
retinoic acids exhibit pleotropic effects, we undertook to prepare
non-retinoid antagonists of mRPE65 to directly determine if
inhibition of this target in and of itself could limit the visual
cycle, establish that RPE65 function is part of the rate-limiting
step in visual pigment regeneration, and lessen the accumulation of
the retinotoxic lipofuscins.
[0919] Two non-retinoid antagonists of mRPE65 were readily designed
and shown to bind potently to mRPE65, a target unique to the visual
cycle. Both TDT and TDH inhibited 11-cis-retinal regeneration after
photobleaching to approximately the same extent as 13-RA. However
unlike 13-RA, both TDT and TDH are directed solely at mRPE65, and
in vivo inhibition results are consistent with this protein being
the operant target. Rodents treated with TDT and TDH accumulate
all-trans-retinyl esters behind the mRPE65 block. This result is
expected, because the all-trans-retinyl esters are converted into
11-cis-retinol more slowly when mRPE65 is inhibited. By comparison,
in the presence of 13-RA, the accumulation of both
all-trans-retinyl and 11-cis-retinyl esters is noted [24]. This
occurs because 13-RA inhibits both mRPE65 and 11-cis-retinol
dehydrogenase [22,23].
[0920] Chronic treatment with TDT and TDH had profound effects on
limiting A.sub.2E accumulation in the animal model of STGD, the
Abca4.sup.-/- mice. TDT, in particular, prevented A.sub.2E
formation by approximately 85% compared to untreated Abca4.sup.-/-
animals, and brought A.sub.2E levels down to approximately those
observed in wt animals of similar age. The relationship between the
extent of inhibition of visual cycle turnover and A.sub.2E
accumulation remains to be explored. It is likely to be non-linear,
at least in part due to the fact that A.sub.2E formation is second
order in all-trans-retinal. Other non-linear effects may operate as
well.
[0921] With respect to the pharmacology of the isoprenoid mRPE65
antagonists, it should be noted that the effects of both TDT and
TDH are substantially more persistent than with 13-RA, probably
because they are more hydrophobic than the retinoic acids. An
increase in hydrophobicity tends to slow down rates of elimination
of drugs. In addition, animals treated with TDT and TDH tolerated
the compounds extremely well, showing no obvious signs of toxicity
and/or distress, even when the compounds were administered every 48
h.
[0922] In conclusion, we have designed and studied specific,
non-retinoid mRPE65 antagonists, which inhibit 11-cis-retinal
regeneration after bleaching, further supporting the hypothesis
that mRPE65 is minimally part of the rate-limiting process in
visual pigment regeneration. The analogs described here will be
useful in a chemical genetic approach to the temporal function of
mRPE65 in visual cycle function visual pigment regeneration.
Similar analogs will be used to probe the function of the
congeneric sRPE65 as it relates to the regulation of the visual
cycle [19]. In addition to analyzing the function of RPE65 in
vitro, the non-retinoid antagonists also profoundly inhibited
lipofuscin A.sub.2E accumulation in the Abca4.sup.-/- mouse model
of macular degeneration. Our studies suggest that these, or similar
molecules, may be efficient and non-toxic candidates as drugs aimed
at preventing the onset of lipofuscin-sensitive forms of macular
degeneration, including STGD and a prevalent form of AMD
(geographic atrophy) leading to visual loss [28].
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[0925] The examples should not be construed as limiting in any way.
The contents of all cited references (including literature
references, issued patents, published patent applications as cited
throughout this application) are hereby expressly incorporated by
reference.
[0926] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents of the specific embodiments described herein.
* * * * *