U.S. patent application number 17/240182 was filed with the patent office on 2021-12-16 for methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid.
The applicant listed for this patent is Bellus Health Inc.. Invention is credited to Mohamed ATFANI, Benoit BACHAND, Abderrahim BOUZIDE, Stephane CIBLAT, Daniel DELORME, Xianqi KONG, Sophie LEVESQUE, David MIGNEAULT, Isabelle VALADE, Xinfu WU.
Application Number | 20210386693 17/240182 |
Document ID | / |
Family ID | 1000005807974 |
Filed Date | 2021-12-16 |
United States Patent
Application |
20210386693 |
Kind Code |
A1 |
KONG; Xianqi ; et
al. |
December 16, 2021 |
METHODS, COMPOUNDS, COMPOSITIONS AND VEHICLES FOR DELIVERING
3-AMINO-1-PROPANESULFONIC ACID
Abstract
The invention relates to methods, compounds, compositions and
vehicles for delivering 3-amino-1-propanesulfonic acid (3APS) in a
subject, preferably a human subject. The invention encompasses
compound that will yield or generate 3APS, either in vitro or in
vivo. Preferred compounds include amino acid prodrugs of 3APS for
use, including but not limited to the prevention and treatment of
Alzheimer's disease
Inventors: |
KONG; Xianqi;
(Dollard-des-Ormeaux, CA) ; ATFANI; Mohamed;
(Laval, CA) ; BACHAND; Benoit; (Saint-Laurent,
CA) ; BOUZIDE; Abderrahim; (Laval, CA) ;
CIBLAT; Stephane; (Montreal, CA) ; LEVESQUE;
Sophie; (Mirabel, CA) ; MIGNEAULT; David;
(Laval, CA) ; VALADE; Isabelle; (Laval, CA)
; WU; Xinfu; (Laval, CA) ; DELORME; Daniel;
(Saint-Lazare, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Bellus Health Inc. |
Laval |
|
CA |
|
|
Family ID: |
1000005807974 |
Appl. No.: |
17/240182 |
Filed: |
April 26, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16857520 |
Apr 24, 2020 |
11020360 |
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17240182 |
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16276941 |
Feb 15, 2019 |
10857109 |
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16857520 |
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15293965 |
Oct 14, 2016 |
10238611 |
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16276941 |
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14246894 |
Apr 7, 2014 |
9499480 |
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15293965 |
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11871639 |
Oct 12, 2007 |
8748656 |
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14246894 |
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60911459 |
Apr 12, 2007 |
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60851039 |
Oct 12, 2006 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 317/40 20130101;
C07H 7/02 20130101; A61K 47/542 20170801; C07C 309/15 20130101;
C07J 9/005 20130101; C07C 309/19 20130101; A61K 47/545 20170801;
A61K 47/554 20170801; C07D 209/20 20130101; C07D 233/64 20130101;
C07D 323/02 20130101; C07K 5/06069 20130101; C12P 13/001 20130101;
C07K 5/0606 20130101; A61K 31/145 20130101; A61K 47/549 20170801;
C07H 15/12 20130101; A61K 47/64 20170801; C07D 285/36 20130101;
C07D 217/24 20130101; C07D 333/24 20130101; C12P 11/00 20130101;
C07K 5/06086 20130101; C07D 291/02 20130101; C07K 5/06052 20130101;
C07C 309/18 20130101; Y02P 20/55 20151101; A61K 47/54 20170801;
C07K 5/0806 20130101; C07K 5/081 20130101; C07K 5/06026 20130101;
C07D 285/38 20130101; C07D 207/16 20130101; C07C 309/24
20130101 |
International
Class: |
A61K 31/145 20060101
A61K031/145; C07D 317/40 20060101 C07D317/40; C07C 309/15 20060101
C07C309/15; C07D 207/16 20060101 C07D207/16; C07D 209/20 20060101
C07D209/20; C07D 217/24 20060101 C07D217/24; C07D 233/64 20060101
C07D233/64; C07D 291/02 20060101 C07D291/02; C07D 333/24 20060101
C07D333/24; C07K 5/062 20060101 C07K005/062; C07K 5/068 20060101
C07K005/068; C07K 5/083 20060101 C07K005/083; C12P 11/00 20060101
C12P011/00; A61K 47/54 20060101 A61K047/54; A61K 47/64 20060101
A61K047/64; C07C 309/18 20060101 C07C309/18; C07C 309/19 20060101
C07C309/19; C07C 309/24 20060101 C07C309/24; C07D 285/36 20060101
C07D285/36; C07D 285/38 20060101 C07D285/38; C07D 323/02 20060101
C07D323/02; C07H 7/02 20060101 C07H007/02; C07H 15/12 20060101
C07H015/12; C07J 9/00 20060101 C07J009/00; C12P 13/00 20060101
C12P013/00 |
Claims
1. A compound selected from the group consisting of: a) a compound
of Formula I: B-L-A (I) wherein B is a pharmacokinetic modulating
moiety, which is optionally also bonded to A directly or indirectly
through a further linking group L; A is a 3-amino-1-propanesulfonic
acid moiety; and L is a cleavable linkage for covalently and
dissociably coupling B to A via the NH.sub.2 group, whereby L can
be a direct bond or additional chemical structure providing a
cleavable linkage, or a pharmaceutically acceptable salt thereof;
b) a compound of Formula I-A: ##STR00151## wherein, R.sup.x and
R.sup.y are independently selected from hydrogen and a protecting
group, wherein R.sup.x and R.sup.y are not both hydrogen; and
L.sup.1 and L.sup.2 are each a cleavable linkage; wherein when
R.sup.x is H, L.sup.1 is absent, and when R.sup.y is H, then
L.sup.2 is absent, or a pharmaceutically acceptable salt thereof;
c) a compound of Formula VII: ##STR00152## wherein, f) a compound
of Formula X: ##STR00153## wherein, R.sup.10 is a residue of a
carbohydrate, a carbohydrate derivative or a carbohydrate-derived
polyol, e.g., a C.sub.5-6 saturated or partially or completely
unsaturated cycloalkyl group, optionally and preferably containing
an --O-- group, which is substituted by 3 to 5 substituents, each
independently selected from --OH, --OAc, --CH.sub.2OH, --OCH.sub.3,
--CH.sub.2OAc and .dbd.O. L is a linking moiety or is absent, e.g.,
an alkyl group, which may be saturated or unsaturated, preferably a
lower alkyl group, which is optionally interrupted by one or more
--O-- and/or --NH-- groups, and is optionally substituted by one or
more .dbd.O, --OH, and/or --NH.sub.2 groups, or a pharmaceutically
acceptable salt thereof; g) a compound of Formula XI: ##STR00154##
wherein, R.sup.11 is a hydrogen or a substituted or unsubstituted
group selected from C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12
alkenyl, C.sub.2-C.sub.12 alkynyl, C.sub.3-C.sub.15 cycloalkyl,
C.sub.3-C.sub.15 heterocycloalkyl, C.sub.6-C.sub.15 aryl,
C.sub.5-C.sub.15 heteroaryl, C(O)R.sup.12, and C(O)OR.sup.13; and
R.sup.12 and R.sup.13 are independently selected from substituted
or unsubstituted C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl,
C.sub.2-C.sub.12 alkynyl, C.sub.3-C.sub.15 cycloalkyl,
C.sub.3-C.sub.15 heterocycloalkyl, C.sub.6-C.sub.15 aryl, and
C.sub.5-C.sub.15 heteroaryl, or a pharmaceutically acceptable salt
thereof; h) a compound of Formula XII: ##STR00155## wherein, D is a
carbonyl, an amino acid residue, or a substituted methylene group;
and R.sup.5 is a substituted or unsubstituted group selected from
C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12
alkynyl, C.sub.3-C.sub.15 cycloalkyl, C.sub.3-C.sub.15
heterocycloalkyl, C.sub.6-C.sub.15 aryl, C.sub.5-C.sub.15
heteroaryl, NH(C.sub.1-C.sub.6 alkyl), N(C.sub.1-C.sub.6
alkyl).sub.2, and C(O)(C.sub.1-C.sub.6 alkyl); R.sup.6 is a
hydrogen or a substituted or unsubstituted group selected from
C(O)NH.sub.2, C(O)NH(C.sub.1-C.sub.6 alkyl), C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, and C(O)(C.sub.1-C.sub.6 alkyl); or R.sup.5 and
R.sup.6 are taken together with the adjacent carbon atom to form a
substituted or unsubstituted C.sub.3-C.sub.12 heterocycloalkyl; M
is selected from the group consisting of oxygen, sulfur, nitrogen
or absent, or a pharmaceutically acceptable salt thereof; d) a
compound of Formula VIII: ##STR00156## wherein, R.sup.7 is a
substituted or unsubstituted group selected from C.sub.1-C.sub.12
alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl,
C.sub.3-C.sub.15 cycloalkyl, C.sub.3-C.sub.15 heterocycloalkyl,
C.sub.6-C.sub.15 aryl, C.sub.5-C.sub.15 heteroaryl,
C.sub.7-C.sub.12 arylalkyl, C.sub.7-C.sub.12 heteroarylalkyl, and
combinations thereof, or a pharmaceutically acceptable salt
thereof; e) a compound of Formula IX: ##STR00157## wherein, R.sup.8
is a substituted or unsubstituted group selected from
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12
alkynyl, C.sub.3-C.sub.15 cycloalkyl, C.sub.3-C.sub.15
heterocycloalkyl, C.sub.6-C.sub.15 aryl, C.sub.5-C.sub.15
heteroaryl; and R.sup.9 is a hydrogen or a substituted or
unsubstituted C(O)(C.sub.1-C.sub.6 alkyl), C(O)NH.sub.2,
C(O)NH(C.sub.1-C.sub.6 alkyl), or C(O)N(C.sub.1-C.sub.6
alkyl).sub.2; or R.sup.8 and R.sup.9 are taken together with the
adjacent carbon atom to form a substituted or unsubstituted
C.sub.3-C.sub.12 heterocycloalkyl, or a pharmaceutically acceptable
salt thereof; X is selected from O, NH, and S, or a
pharmaceutically acceptable salt thereof; i) a compound of Formula
XIII: ##STR00158## wherein, R.sup.15 and R.sup.16 are independently
selected from a hydrogen or a substituted or unsubstituted group
selected from C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl,
C.sub.2-C.sub.12 alkynyl, C.sub.3-C.sub.15 cycloalkyl,
C.sub.3-C.sub.15 heterocycloalkyl, C.sub.6-C.sub.15 aryl, and
C.sub.5-C.sub.15 heteroaryl, or a pharmaceutically acceptable salt
thereof; and j) a compound of Formula I-P:
A-(L.sup.x-A).sub.p-L.sup.x-A (I-P) wherein: A is
3-amino-1-propanesulfonic acid moiety; L.sup.x is a cleavable
linkage for covalently and dissociably coupling together two
adjacent 3APS moieties, and p is 0, or an integer number which may
vary from 1 to 5, e.g. 2, 3, 4, or 5; or a pharmaceutically
acceptable salt thereof.
2. The compound of claim 1, wherein the compound of Formula XII is
a compound of Formula XII-A: ##STR00159## wherein, R.sup.14 is a
substituted or unsubstituted group selected from C.sub.1-C.sub.12
alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl,
C.sub.3-C.sub.15 cycloalkyl, C.sub.3-C.sub.15 heterocycloalkyl,
C.sub.6-C.sub.15 aryl, C.sub.5-C.sub.15 heteroaryl, or a
pharmaceutically acceptable salt thereof.
3. The compound of claim 1, wherein said compound of Formula I-P is
selected from the group consisting of: a) a compound of Formula
I-P2: L.sup.y(A).sub.m (I-P2) wherein, m is an integer from 2 to 5;
A is 3-amino-1-propanesulfonic acid moiety; L.sup.y is a
multivalent carrier moiety for covalently and dissociably coupling
from two to five A moieties, either at the amino or sulfonic acid
end of A, or a pharmaceutically acceptable salt thereof; b) a
compound of Formula I-C: ##STR00160## wherein, L.sup.3 is bivalent
linker which connects two molecules of 3APS at their amino groups
either using the same or different linkages as defined herein,
including, but not limited to, amide linkage and carbamate linkage,
or a pharmaceutically acceptable salt thereof; c) a compound of
Formula I-D: ##STR00161## wherein, L.sup.4 is a bivalent linker
which connects two molecules of 3APS at their sulfonic acid groups
either using the same or different linkages as defined herein,
including, but not limited to, ester linkage or anhydride linkage
where X is oxygen, or sulfonamide linkage where X is nitrogen (NH,
or NR), or thiosulfonate linkage where X is sulfur, P is hydrogen
or a N-protecting group, or a pharmaceutically acceptable salt
thereof; and d) a compound of Formula I-E: ##STR00162## wherein,
L.sup.5 is a bivalent linker which connects two molecules of 3APS,
at amino group in one 3APS using a linkage as defined in Formula
I-C, and at sulfonic acid group in the other 3APS using a linkage
as defined in Formula I-D, P is hydrogen or a N-protecting group as
defined herein, or a pharmaceutically acceptable salt thereof.
4. The compound of claim 1, wherein said compound is selected from
the group consisting of: ##STR00163## ##STR00164## ##STR00165##
##STR00166## ##STR00167## ##STR00168## ##STR00169## ##STR00170## or
a pharmaceutically acceptable salt thereof.
5. A method for treating Alzheimer's disease, mild cognitive
impairment, Down's syndrome, Hereditary Cerebral Hemorrhage with
Amyloidosis of the Dutch-Type, cerebral amyloid angiopathy, a
degenerative dementia, a dementia of mixed vascular and
degenerative origin, dementia associated with Parkinson's disease,
dementia associated with progressive supranuclear palsy, dementia
associated with cortical basal degeneration, or diffuse Lewy body
type of Alzheimer's disease, comprising administering a
therapeutically effective amount of a compound of claim 1 to a
human subject in need thereof.
6. The method of claim 5, which is for treating Alzheimer's
disease, mild cognitive impairment, cerebral amyloid angiopathy, or
degenerative dementia.
7. The method of claim 6, which is for treating Alzheimer's
disease.
8. The method of claim 5 wherein the compound is administered
intratracheally, intranasally, ontologically, rectally, vaginally,
or orally.
9. A pharmaceutical composition comprising a compound of claim 1
together with a pharmaceutically acceptable carrier.
10. The pharmaceutical composition of claim 9 which is suitable for
oral administration.
11. The pharmaceutical composition of claim 9, which is in the form
of a hard shell gelatin capsule, soft shell gelatin capsule,
cachet, pill, tablet, lozenge, powder, granule, pellet, dragee,
which is optionally enteric coated, a solution, an aqueous liquid
suspension, a non-aqueous liquid suspension, an oil-in-water liquid
emulsion, a water-in-oil liquid emulsion, an elixir, a syrup, or a
pastille.
12. A process for converting a compound of claims 1 to
3-amino-1-propanesulfonic acid (3APS) comprising contacting said
compound with plasma, blood and/or brain cells whereby said
compound is metabolized to 3APS.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 15/293,965, filed Oct. 14, 2016, now allowed,
which is a continuation of U.S. patent application Ser. No.
14/246,894, filed Apr. 7, 2014, now U.S. Pat. No. 9,499,480, which
is a continuation of U.S. patent application Ser. No. 11/871,639,
filed Oct. 12, 2007, now U.S. Pat. No. 8,748,656, which claims
priority to U.S. Provisional Patent Application No. 60/851,039,
filed Oct. 12, 2006, and U.S. Provisional Patent Application No.
60/911,459, filed Apr. 12, 2007, all of which are incorporated
herein by reference in their entireties.
FIELD OF THE INVENTION
[0002] The invention relates to methods, compounds, compositions
and vehicles for delivering 3-amino-1-propanesulfonic acid (3APS)
in a subject, preferably a human subject. The invention encompasses
compounds that will yield or generate 3APS, either in vitro or in
vivo. Preferred compounds include amino acid prodrugs of 3APS for
use, including but not limited to, the prevention and treatment of
Alzheimer's disease.
BACKGROUND OF THE INVENTION
[0003] Alzheimer's disease (AD) is a progressive degenerative
disease of the brain primarily associated with aging. Prevalence of
AD in the United States in 2000 was close to 4.5 Million. It was
estimated that about one in ten individuals over 65 and nearly half
of those over 85 are affected by Alzheimer's disease. Approximately
360,000 patients will be diagnosed with AD each year in the United
States alone.
[0004] Clinical presentation of AD is characterized by loss of
memory, cognition, reasoning, judgment, and orientation. As the
disease progresses, motor, sensory, and linguistic abilities are
also affected until there is global impairment of multiple
cognitive functions. These cognitive losses occur gradually, but
typically lead to severe impairment and eventual death in the range
of four to twelve years.
[0005] Alzheimer's disease is characterized by two major pathologic
observations in the brain: neurofibrillary tangles and beta amyloid
(or neuritic) plaques, comprised predominantly of an aggregate of a
peptide fragment know as A13. Individuals with AD exhibit
characteristic beta-amyloid deposits in the brain (beta amyloid
plaques) and in cerebral blood vessels (beta amyloid angiopathy) as
well as neurofibrillary tangles. Neurofibrillary tangles occur not
only in Alzheimer's disease but also in other dementia-inducing
disorders.
[0006] 3-amino-1-propanesulfonic acid (3APS, Tramiprosate,
Alzhemed.TM.) is a promising investigational product candidate for
the treatment of Alzheimer's disease that is currently in Phase III
clinical trials in North America and Europe (Wright, T. M., Drugs
of Today (2006), 42(5): 291-298). This product is developed by
Neurochem Inc. (Laval, QC, Canada) and it is believed to act by
reducing the deposition and/or load of amyloid in the brain through
its binding to soluble A.beta. peptide. For increasing the
therapeutic effectiveness of 3APS, it would be desirable to
increase bioavailability, stability and/or crossing the blood brain
barrier of 3APS. These and other needs can be satisfied by the
disclosure herein of a prodrug form of 3-amino-1-propanesulfonic
acid (3APS), pharmaceutical compositions and uses thereof to treat
various medical disorders.
[0007] Previous metabolic stability studies had demonstrated that
there was no in vitro metabolism of 3APS. Those studies include:
3APS metabolic stability in pooled human hepatocytes, human, rat
and dog liver microsomes, human intestinal microflora, pooled human
liver cytosol, and human arylamine N-acetyltransferase (See
Examples 4 and 5).
SUMMARY OF THE INVENTION
[0008] Surprisingly, it has now been found that 3APS is metabolized
both in vitro and in vivo. Indeed, as described in more detail
hereinafter, recent in vivo studies indicate extensive metabolism,
particularly first-pass and/or systemic metabolism of 3APS. Three
potential metabolites were identified from at least one type of
biological species: 2-carboxyethanesulfonic acid,
3-hydroxy-1-propanesulfonic acid and
3-acetylamino-1-propanesulfonic acid. Further studies demonstrated
that 2-carboxyethanesulfonic acid was the only major metabolite of
3APS in mice, rats, dogs and humans.
##STR00001##
[0009] Without wishing to be bound by theory, it is hypothesized
that metabolism of 3APS is mainly caused by a transaminase and/or
monoamine oxidase that generates 2-carboxyethanesulfonic acid as
the main metabolite and 3-hydroxy-1-propanesulfonic acid as a minor
metabolite. N-acetyl-3-aminopropanesulfonic acid is another
possible minor metabolite and it is believed to be produced by an
enzyme that acetylates 3APS. These hypotheses are supported by in
vitro experiments (see, for example, Example 5) showing that the
conversion of 3APS to 2-carboxyethanesulfonic acid in primary
neuron culture media was significantly inhibited by vigabatrin, a
classic GABA transaminase inhibitor. Nialamide, a monoamine oxidase
inhibitor, also reduced the formation of 2-carboxyethanesulfonic
acid (from 3APS) but to a lesser extent.
[0010] Accordingly, an aspect of the invention concerns compounds
and compositions that can deliver 3APS by minimizing the
metabolism, e.g., first-pass metabolism, associated with that drug,
and more particularly compounds that would block or protect the
amino group of 3APS such that it avoids metabolism, e.g., by
transaminases and/or monoamine oxidases.
[0011] The invention includes methods, compounds, compositions and
vehicles for delivering in a subject, preferably a human subject,
3-amino-1-propanesulfonic acid, or salts thereof.
3-Amino-1-propanesulfonic acid (also named 3APS, Tramiprosate,
Alzhemed.TM.) has the structure:
##STR00002##
[0012] According to an aspect, the present invention relates to
compounds or compositions that will yield or generate 3APS after
administered in a subject. In one embodiment, the compound that
will yield or generate 3APS is an amino acid prodrug of 3APS. In
another embodiment, the compound that will yield or generate 3APS
is a carbamate prodrug of 3APS. In another embodiment, the compound
that will yield or generate 3APS is an amide prodrug of 3APS. In
another embodiment, the compound that will yield or generate 3APS
is a carbohydrate-derived prodrug of 3APS. In another embodiment,
the compound that will yield or generate 3APS is a N-hydroxy
prodrug. In another embodiment, the compound that will yield or
generate 3APS is a cyclic double-protected prodrug. In further
embodiment, the compound that yields or generates 3APS is a 3APS
polymer (e.g. a molecule composed of two or more molecules of 3APS
linked together). In further embodiment, the compound that yields
or generates 3APS is a gemini dimer of 3APS. In certain
embodiments, the amino acid prodrugs of 3APS that are capable of
yielding or generating, either in vitro or in vivo, 3APS have one
of the general or specific formulae or structures disclosed herein.
The present invention encompasses these compounds, pharmaceutical
compositions containing these compounds, and methods employing such
compounds or compositions in the treatment of various medical
disorders such as Alzheimer's disease.
[0013] The present invention also relates to pharmaceutical
compositions comprising a compound of the present invention.
[0014] The present invention further relates to a method for
increasing the therapeutic effectiveness of 3APS comprising
administering to a subject, preferably a human subject, an
effective amount of a prodrug of the present invention.
[0015] The present invention also provides processes for converting
compounds of the invention to 3APS. The conversion and/or
generation of 3APS involves contacting any of the compounds of the
invention, e.g., with blood, plasma and/or brain cells. The
conversion can occur in vitro or in viva The conversion may also
occur in the presence of enzymes capable of cleaving amine bonds,
such as peptidases, or other enzymes appropriate for other
structures herein, including those found in the blood, plasma
and/or brain.
[0016] The invention also provides the use of a compound according
to the invention for the manufacture of a medicament. The invention
also provides the use of a compound of the invention for the
treatment or prevention of Alzheimer's disease, mild cognitive
impairment, Down's syndrome, Hereditary Cerebral Hemorrhage with
Amyloidosis of the Dutch-Type, cerebral amyloid angiopathy, other
degenerative dementias, dementias of mixed vascular and
degenerative origin, dementia associated with Parkinson's disease,
dementia associated with progressive supranuclear palsy, dementia
associated with cortical basal degeneration, or diffuse Lewy body
type of Alzheimer's disease. The invention also provides methods
for the treatment or prevention of the aforementioned diseases
comprising administration of a therapeutically effective amount of
a compound of the invention or a composition comprising the same,
to a subject, preferably a human subject, in need thereof. More
preferably, the disease is Alzheimer's disease. Accordingly, a
related aspect of the invention relates to the prevention and/or
treatment of Alzheimer's disease in a human subject by
administering an effective amount of a compound or composition of
the present invention to a human subject in need thereof.
[0017] In a further embodiment the invention includes
administration of 3APS via or under the mucous membranes, the nose
(intranasally), mouth, or eye, e.g., by nasal spray, chewing gum,
or eye drops, via the ear, e.g., by eardrops, by the use of an
implant, rectally, e.g., by a suppository or enema, vaginally,
e.g., by a cream or lotion, or by the respiratory system, e.g., by
inhalation, intranasally or intratracheally.
[0018] The invention in further aspects includes the administration
of compounds of the invention via any mode and/or vehicle,
including all modes and/or vehicles disclosed herein, e.g., the
administration of the prodrugs of 3APS via the nose, mucous
membranes, transdermally, via a patch, etc.
[0019] This invention in various aspects relates to the following
numbered aspects:
[0020] Aspect 1. A compound of the Formula I:
B-L-A (I)
wherein B is a pharmacokinetic modulating moiety, which is
optionally also bonded to A directly or indirectly through a
further linking group L; A is a 3-amino-1-propanesulfonic acid
moiety (i.e., 3APS bound to L-B), and L is a cleavable linkage for
covalently and dissociably coupling B to A (preferably and
typically via the NH.sub.2 group), or is absent, whereby L can be a
direct bond or additional chemical structure providing a cleavable
linkage, or a pharmaceutically acceptable salt or solvate
thereof.
[0021] Aspect 2. The compound according to aspect 1, wherein
L is a linkage that when metabolized or hydrolyzed either in vitro
or in vivo produces 3APS, and/or B is a moiety that increases the
therapeutic bio-distribution of 3APS upon administration of the
compound of formula Ito a human subject.
[0022] Aspect 3. The compound according to aspect 1, wherein B is a
3-amino-1-propanesulfonic acid moiety.
[0023] Aspect 4. A compound according to aspect 1, wherein
B is an amino acid or a peptide, and L is a hydrolyzable
linkage.
[0024] Aspect 5. A compound according to aspect 1, which is a
compound of formula (I), (I-A), (I-C), (I-D), (I-E), (I-P), (I-P2),
(II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI),
(XII), (XII-A) or (XIII), which are set forth hereinafter, or a
pharmaceutically acceptable salt thereof.
[0025] Aspect 6. A pharmaceutical composition comprising a compound
of aspect 1 and a pharmaceutically acceptable vehicle.
[0026] Aspect 7. A method for treating or preventing Alzheimer's
disease, mild cognitive impairment Down's syndrome, Hereditary
Cerebral Hemorrhage with Amyloidosis of the Dutch-Type, cerebral
amyloid angiopathy, a degenerative dementia, a dementia of mixed
vascular and degenerative origin, dementia associated with
Parkinson's disease, dementia associated with progressive
supranuclear palsy, dementia associated with cortical basal
degeneration, or diffuse Lewy body type of Alzheimer's disease
comprising administering a therapeutically effective amount of a
compound of aspect 1 to a human subject in need thereof.
[0027] Aspect 8. A process for converting a compound of aspect 1 to
3APS comprising contacting said compound with an enzyme which
metabolizes said compound to 3APS in vitro or in vivo.
[0028] Aspect 9. A process according to aspect 8, comprising
contacting said compound with plasma, blood and/or brain cells.
[0029] Aspect 10. A method for increasing the therapeutic
bio-distribution of 3APS in a human subject, comprising lessening
metabolism of 3APS, e.g., first pass metabolism, which occurs when
3APS is administered to a human subject.
[0030] Aspect 11. A method for reducing side effects of 3APS in a
human subject (e.g., reducing or preventing gastrointestinal
intolerance), comprising lessening metabolism of 3APS, e.g., first
pass metabolism, which occurs when 3APS is administered to a human
subject.
[0031] Aspect 12. A method according to aspect 10, wherein 3APS is
administered in the form of a prodrug of 3APS which yields or
generates 3APS after being administered to said human subject.
[0032] Aspect 13. A method according to aspect 10, wherein the
prodrug is a compound of the Formula I:
B-L-A (I)
wherein B is a pharmacokinetic modulating moiety, which is
optionally also bonded to A directly or indirectly through a
further linking group L; A is a 3-amino-1-propanesulfonic acid
moiety (i.e., 3APS bound to L-B), and L is a cleavable linkage for
covalently and dissociably coupling B to A (preferably and
typically via the NH.sub.2 group), or is absent, whereby L can be a
direct bond or additional chemical structure providing a cleavable
linkage, or a pharmaceutically acceptable salt or solvate
thereof.
[0033] Aspect 14. A method according to aspect 10, wherein 3APS is
administered through the respiratory system, intratracheally,
intranasally, via or under a mucous membrane, via the ear,
rectally, or vaginally, or by an implant, spray, nasal spray,
chewing gum, eye drop, eardrop, suppository, enema, or vaginal
cream or lotion.
[0034] Aspect 15. The method of aspect 10, wherein the
bioavailability of 3APS, AUC of 3APS, brain levels of 3APS, CSF
levels of 3APS, C.sub.max of 3APS, T.sub.max of 3APS, and/or
bio-absorption of 3APS is increased.
[0035] Aspect 16. A method according to aspect 10, wherein
Alzheimer's disease is treated or prevented.
[0036] Aspect 17. A method according to aspect 10, wherein the
effective therapeutic level of 3APS in a selected human tissue is
increased.
[0037] Aspect 18. A method according to aspect 17, which increases
the level of 3APS in the brain of said human subject.
[0038] Aspect 19. A method according to aspect 10, which increases
the therapeutic effectiveness of 3APS.
[0039] Aspect 20. A method according to aspect 10, which lessens
the first pass metabolism of 3APS.
[0040] Aspect 21. A method according to aspect 10, which reduces
the side effects of 3APS.
[0041] Aspect 22. A method according to aspect 10, wherein the oral
AUC of 3APS is increased by at least 20%.
[0042] Aspect 23. A method for increasing the therapeutic
bio-distribution of 3APS in a human subject, comprising
administering 3APS in the form of a prodrug or in the form of a
gemini dimer of 3APS.
[0043] Aspect 24. A method for increasing the therapeutic
bio-distribution of 3APS in a human subject, comprising
administering 3APS non-orally or non-enterally.
[0044] Aspect 25. A method according to aspect 10, wherein 3APS is
delivered using a route (transdermally, S.C., intranasally, etc.)
or vehicle (patch, implant, spray, formulation, etc.) which
minimizes hepatic first-pass metabolism of 3APS.
[0045] Aspect 26. A compound according to aspect 1, wherein said
cleavable linkage is selected for yielding or generating 3APS or a
derivative of 3APS, either in vitro or in vivo, e.g., wherein the
linkage is cleavable hydrolytically or enzymatically.
[0046] Aspect 27. A compound according to aspect 1, wherein said
pharmacokinetic modulating moiety is selected for increasing the
therapeutic bio-distribution of 3APS upon administration of the
compound of formula Ito a human subject.
[0047] Aspect 28. A prodrug of the Formula I:
B-L-A (I)
wherein B is a pharmacokinetic modulating moiety, which is
optionally also bonded to A directly or indirectly through a
further linking group L; A is a 3-amino-1-propanesulfonic acid
moiety (i.e., 3APS bound to L-B), and L is a cleavable linkage for
covalently and dissociably coupling B to A (preferably and
typically via the NH.sub.2 group), or is absent, whereby L can be a
direct bond or additional chemical structure providing a cleavable
linkage, or a pharmaceutically acceptable salt, metabolite or
solvate thereof,
[0048] wherein the metabolite of said prodrug can be 3APS and/or
other metabolites, including, but not limited to, metabolites
identified elsewhere herein, e.g., the examples.
[0049] Additional objects, advantages and features of the present
invention will become more apparent upon reading of the following
non-restrictive description of preferred embodiments which are
exemplary and should not be interpreted as limiting the scope of
the invention.
I. Definitions
[0050] All technical and scientific terms used herein have the same
meaning as commonly understood by one ordinary skilled in the art
to which the invention pertains. For convenience, the meaning of
certain terms and phrases used herein are provided below.
[0051] To the extent the definitions of terms in the publications,
patents, and patent applications incorporated herein by reference
are contrary to the definitions set forth in this specification,
the definitions in this specification control. The section headings
used herein are for organizational purposes only, and are not to be
construed as limiting the subject matter disclosed.
[0052] It should be noted that, the singular forms "a", "an", and
"the" include plural referents unless the content clearly dictates
otherwise. Thus, for example, reference to a composition containing
"a compound" includes a mixture of two or more compounds. It should
also be noted that the term "or" is generally employed in its sense
including "and/or" unless the content clearly dictates
otherwise.
[0053] The chemical structures herein are drawn according to the
conventional standards known in the art. Thus, where an atom, such
as a carbon atom, as drawn appears to have an unsatisfied valency,
then that valency is assumed to be satisfied by a hydrogen atom
even though that hydrogen atom is not necessarily explicitly drawn.
Hydrogen atoms should be inferred to be part of the compound.
[0054] The symbol "--" in general represents a bond between two
atoms in the chain. Thus
CH.sub.3--O--CH.sub.2--CH(R.sub.i)--CH.sub.3 represents a
2-substituted-1-methoxypropane compound. In addition, the symbol
"--" represents the point of attachment of the substituent to a
compound. Thus for example aryl(C.sub.1-C.sub.6)--alkyl indicates
an arylalkyl group, such as benzyl, attached to the compound at the
alkyl moiety.
[0055] Where multiple substituents are indicated as being attached
to a structure, it is to be understood that the substituents can be
the same or different. Thus for example "R.sub.m optionally
substituted with 1, 2 or 3 R.sub.q groups" indicates that R.sub.m
is substituted with 1, 2, or 3 R.sub.q groups where the R.sub.q
groups can be the same or different.
[0056] As used herein, the term "Compounds of the present
invention" and equivalent expressions refers to compounds mentioned
herein as being useful for at least one purpose of the invention,
e.g., those encompassed by structural Formulae such as (I), (I-A),
(I-C), (I-D), (I-E), (I-P), (I-P2), (II), (III), (IV), (V), (VI),
(VII), (VIII), (IX), (X), (XI), (XII), (XII-A) and (XIII), and
includes specific compounds mentioned herein such as A1 to A35, C1
to C26, B1 to B14, H1 to H4, G1 to G11, S1 to S14 and D1 to D8,
etc., as well as their pharmaceutically acceptable salts and
solvates. Embodiments herein may exclude one or more of the
compounds of the invention. Compounds may be identified either by
their chemical structure and/or chemical name. When the chemical
structure and chemical name conflict, the chemical structure is
determinative of the identity of the compound. The compounds
described herein may contain one or more chiral centers and/or
double bonds and therefore, may exist as stereoisomers, such as
double-bond isomers (i.e., geometric isomers), enantiomers, or
diastereomers. Accordingly, the chemical structures disclosed
herein encompass all possible enantiomers and stereoisomers of the
illustrated compounds including the stereoisomerically pure form
(e.g., geometrically pure, enantiomerically pure, or
diastereomerically pure) and enantiomeric and stereoisomeric
mixtures. Enantiomeric and stereoisomeric mixtures can be resolved
into their component enantiomers or stereoisomers using separation
techniques or chiral synthesis techniques well known to the skilled
artisan, e.g., chiral chromatography (such as chiral HPLC),
immunoassay techniques, or the use of covalently (such as Mosher's
esters) and non-covalently (such as chiral salts) bound chiral
reagents to respectively form a diastereomeric mixture which can be
separated by conventional methods, such as chromatography,
distillation, crystallization or sublimation, the chiral salt or
ester is then exchanged or cleaved by conventional means, to
recover the desired isomers. The compounds may also exist in
several tautomeric forms including the enol form, the keto form,
and mixtures thereof. Accordingly, the chemical structures depicted
herein encompass all possible tautomeric forms of the illustrated
compounds. The disclosed compounds also include isotopically
labeled compounds where one or more atoms have an atomic mass
different from the atomic mass most abundantly found in nature.
Examples of isotopes that may be incorporated into the compounds of
the present invention include, but are not limited to, .sup.2H (D),
.sup.3H (T), .sup.11C, .sup.13C, .sup.14C, .sup.15N.sub., .sup.18O,
.sup.17O etc. Compounds may exist in unsolvated forms as well as
solvated forms, including hydrated forms. In general, compounds may
be hydrated or solvated. Certain compounds may exist in multiple
crystalline or amorphous forms. In general, all physical forms are
equivalent for the uses contemplated herein and are intended to be
within the scope of the present invention. Further, when partial
structures of the compounds are illustrated, brackets or
equivalents indicate the point of attachment of the partial
structure to the rest of the molecule.
[0057] The term "prodrug" and equivalent expressions refer to
agents which can be converted in vitro or in vivo directly or
indirectly to an active form (see, e.g., R. B. Silverman, 1992,
"The Organic Chemistry of Drug Design and Drug Action," Academic
Press, Chap. 8; Bundgaard, Hans; Editor. Neth. (1985), "Design of
Prodrugs". 360 pp. Elsevier, Amsterdam; Stella, V.; Borchardt, R.;
Hageman, M.; Oliyai, R.; Maag, H.; Tilley, J. (Eds.) (2007),
"Prodrugs: Challenges and Rewards, XVIII," 1470 p. Springer).
Prodrugs can be used to alter the biodistribution (e.g., to allow
agents which would not typically enter the reactive site of the
protease) or the pharmacokinetics for a particular agent. A wide
variety of groups have been used to modify compounds to form
prodrugs, for example, esters, ethers, phosphates, etc. When the
prodrug is administered to a subject, the group is cleaved,
enzymatically or non-enzymatically, reductively, oxidatively, or
hydrolytically, or otherwise to reveal the active form. As used
herein, "prodrug" includes pharmaceutically acceptable salts
thereof, or pharmaceutically acceptable solvates as well as
crystalline forms of any of the foregoing. Prodrugs are frequently,
although not necessarily, pharmacologically inactive until
converted to the parent drug.
[0058] The term "gemini dimer" and equivalent expressions refer to
a synthetic compound comprising at least two moieties of the same
agent or drug coupled together. For background on gemini dimers,
see: Hammell D C, Hamad M, Vaddi H K, Crooks P A, Stinchcomb A L. A
duplex "Gemini" prodrug of naltrexone for transdermal delivery. J
Control Release. 2004. 97(2):283-90. In preferred embodiment, the
gemini dimers of the invention are made of two linked 3APS
molecules that may be converted in vitro or in vivo directly or
indirectly to release at least one, preferably two,
pharmaceutically active 3APS molecules.
[0059] The term "carbamate" refers to an oxycarbonyl residue
(--OC(O)--) linked to an amino group to form a group comprising a
(--OC(O)N(or NH)--) radical The carbamate group can be secondary
(NH) or tertiary (N). This term is further defined in Section
II-B(a).
[0060] The term "amide" refers to an organic compound containing a
carbonyl (--C(O)--) attached to an amine group to form a group
comprising the radical (--C(O)N(or NH)--). The amide group can be
secondary (NH) or tertiary (N). This term is further defined in
Section II-A. The term "non-amino acid amide" refers to an amide
group where the carbonyl (--C(O)--) does not form part of an amino
acid residue. This term is further defined in Section II-B(b).
[0061] The term "carbohydrate-derived" refers to compounds where
the group attached to, for example, 3APS, is an organic group that
is or is derived from a polyhydroxy aldehyde, polyhydroxy ketone,
or a polyol, can change to such group on simple chemical
transformations, such as hydrolysis, oxidation, or reduction. These
groups include, for example, sugars, starches, celluloses, and
gums. This term is further defined in Section II-C. The term
"N-hydroxy-derived" refers to compounds containing a hydroxy or
hydroxy-derived group (e.g. alkoxy, benzyloxy, phenoxy, acyloxy,
and the like) to form an (RO--N(or NH)--). This term is further
defined in Section II-D(a).
[0062] The term "cyclic double-protected" refers to compounds
wherein a protecting group in linked to both the amine and the
sulfonic acid of 3APS. This term is further defined in Section
II-D(b).
[0063] The term "ester" refers to compounds that can be represented
by the formula RCOOR (carboxylic ester) or the formula RSO.sub.3R'
(sulfonate ester)', where the group R can be, for example 3APS or
the 3-aminopropane part thereof, and the group R' can be another
organic group. These compounds are usually respectively formed by
the reaction between a carboxylic or a sulfonic acid and an alcohol
usually with the elimination of water.
[0064] The term "amino acid" generally refers to an organic
compound comprising both a carboxylic acid group and an amine
group. The term "amino acid" includes both "natural" and
"unnatural" or "non-natural" amino acids. Additionally, the term
amino acid includes O-alkylated or N-alkylated amino acids, as well
as amino acids having nitrogen or oxygen-containing side chains
(such as Lys, Orn, or Ser) in which the nitrogen or oxygen atom has
been acylated or alkylated. Amino acids may be pure L or D isomers
or mixtures of L and D isomers, including racemic mixtures. In
general, amino acids are represented by the residue of Formula
V.
[0065] The term "natural amino acid" and equivalent expressions
refer to L-amino acids commonly found in naturally occurring
proteins. Examples of natural amino acids include, without
limitation, alanine (Ala), cystein (Cys), aspartic acid (Asp),
glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine
(His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine
(Met), asparagine (Asp), proline (Pro), glutamine (Gln), arginine
(Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan
(Trp), tyrosine (Tyr), .beta.-alanine (.beta.-ALA), and
.gamma.-aminobutyric acid (GABA).
[0066] The term "unnatural amino acid" refers to any derivative of
a natural amino acid including D forms, and .alpha.- and
.beta.-amino acid derivatives. The terms "unnatural amino acid" and
"non-natural amino acid" are used interchangeably herein and are
meant to include the same moieties. It is noted that certain amino
acids, e.g., hydroxyproline, that are classified as a non-natural
amino acid herein, may be found in nature within a certain organism
or a particular protein. Amino acids with many different protecting
groups appropriate for immediate use in the solid phase synthesis
of peptides are commercially available. In addition to the twenty
most common naturally occurring amino acids, the following examples
of non-natural amino acids and amino acid derivatives may be used
according to the invention (common abbreviations in parentheses):
2-aminoadipic acid (Aad), 3-aminoadipic acid (.beta.-Aad),
2-aminobutyric acid (2-Abu), .alpha.,.beta.-dehydro-2-aminobutyric
acid (8-AU), 1-aminocyclopropane-1-carboxylic acid (ACPC),
aminoisobutyric acid (Aib), 3-aminoisobutyric acid (.beta.-Aib),
2-amino-thiazoline-4-carboxylic acid, 5-aminovaleric acid (5-Ava),
6-aminohexanoic acid (6-Ahx), 2-aminoheptanoic acid (Ahe),
8-aminooctanoic acid (8-Aoc), 11-aminoundecanoic acid (11-Aun),
12-aminododecanoic acid (12-Ado), 2-aminobenzoic acid (2-Abz),
3-aminobenzoic acid (3-Abz), 4-aminobenzoic acid(4-Abz),
4-amino-3-hydroxy-6-methylheptanoic acid (Statine, Sta),
aminooxyacetic acid (Aoa), 2-aminotetraline-2-carboxylic acid
(ATC), 4-amino-5-cyclohexyl-3-hydroxypentanoic acid (ACHPA),
para-aminophenylalanine (4-NH.sub.2-Phe), 2-aminopimelic acid
(Apm), biphenylalanine (Bip), para-bromophenylalanine (4-Br-Phe),
ortho-chlorophenylalanine (2-CI-Phe), meta-chlorophenylalanine
(3-CI-Phe), para-chlorophenylalanine (4-CI-Phe),
meta-chlorotyrosine (3-CI-Tyr), para-benzoylphenylalanine (Bpa),
tert-butylglycine (TLG), cyclohexylalanine (Cha), cyclohexylglycine
(Chg), desmosine (Des), 2,2-diaminopimelic acid (Dpm),
2,3-diaminopropionic acid (Dpr), 2,4-diaminobutyric acid (Dbu),
3,4-dichlorophenylalanine (3,4-C1.sub.2-Phe),
3,4-diflurorphenylalanine (3,4-F.sub.2-Phe), 3,5-diiodotyrosine
(3,5-I.sub.2-Tyr), N-ethylglycine (EtGly), N-ethylasparagine
(EtAsn), ortho-fluorophenylalanine (2-F-Phe),
meta-fluorophenylalanine (3-F-Phe), para-fluorophenylalanine
(4-F-Phe), meta-fluorotyrosine (3-F-Tyr), homoserine (Hse),
homophenylalanine (Hfe), homotyrosine (Htyr), hydroxylysine (Hyl),
allo-hydroxylysine (aHyl), 5-hydroxytryptophan (5-OH-Trp), 3- or
4-hydroxyproline (3- or 4-Hyp), para-iodophenylalanine (4-I-Phe),
3-iodotyrosine (3-I-Tyr), indoline-2-carboxylic acid (Idc),
isodesmosine (Ide), allo-isoleucine (a-lle), isonipecotic acid
(Inp), N-methylisoleucine (Melle), N-methyllysine (MeLys),
meta-methyltyrosine (3-Me-Tyr), N-methylvaline (MeVal),
1-naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal),
para-nitrophenylalanine (4-NO.sub.2-Phe), 3-nitrotyrosine
(3-NO.sub.2-Tyr), norleucine (Nle), norvaline (Nva), ornithine
(Orn), ortho-phosphotyrosine (H.sub.2PO.sub.3-Tyr),
octahydroindole-2-carboxylic acid (Oic), penicillamine (Pen),
pentafluorophenylalanine (F.sub.5-Phe), phenylglycine (Phg),
pipecolic acid (Pip), propargylglycine (Pra), pyroglutamic acid
(PGLU), sarcosine (Sar), tetrahydroisoquinoline-3-carboxylic acid
(Tic), thienylalanine, and thiazolidine-4-carboxylic acid
(thioproline, Th).
[0067] As used herein, the term "acyclic" refers to an organic
moiety without ring system.
[0068] The term "aliphatic group" includes organic moieties
characterized by straight or branched-chains, typically having
between 1 and 15 carbon atoms. Aliphatic groups include non cyclic
alkyl groups, alkenyl groups, and alkynyl groups. As used herein,
the term "alkyl" refers to saturated hydrocarbons having from one
to twelve carbon atoms, including linear, branched, and cyclic
alkyl groups. Examples of alkyl groups include, without limitation,
methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl,
decyl, isopropyl, tert-butyl, sec-butyl, isobutyl, cyclopropyl,
cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. The
term alkyl includes both unsubstituted alkyl groups and substituted
alkyl groups. The term "C.sub.1-C.sub.nalkyl", wherein n is an
integer from 2 to 12, refers to an alkyl group having from 1 to the
indicated "n" number of carbon atoms.
[0069] As used herein, the term "alkenyl" refers to unsaturated
hydrocarbons having from two to twelve carbon atoms, including
linear, branched, and cyclic non aromatic alkenyl groups, and
comprising between one to six carbon-carbon double bond. Examples
of alkenyl groups include, without limitation, vinyl, allyl,
1-propen-2-yl, 1-buten-3-yl, 1-buten-4-yl, 2-buten-4-yl,
1-penten-5-yl, 1,3-pentadien-5-yl, cyclopentenyl, cyclohexenyl,
ethylcyclopentenyl, ethylcylohexenyl, and the like. The term
alkenyl includes both unsubstituted alkenyl groups and substituted
alkenyl groups. The term "C.sub.2-C.sub.nalkenyl", wherein n is an
integer from 3 to 12, refers to an alkenyl group having from 2 to
the indicated "n" number of carbon atoms.
[0070] As used herein, the term "alkynyl" refers to unsaturated
hydrocarbons having from two to twelve carbon atoms, including
linear, branched, and cyclic non aromatic alkynyl groups, and
comprising between one to six carbon-carbon triple bond. Examples
of alkynyl groups include, without limitation, ethynyl,
1-propyn-3-yl, 1-butyn-4-yl, 2-butyn-4-yl, 1-pentyn-5-yl,
1,3-pentadiyn-5-yl, and the like. The term alkynyl includes both
unsubstituted alkynyl groups and substituted alkynyl groups. The
term "C.sub.2-C.sub.nalkynyl", wherein n is an integer from 3 to
12, refers to an alkynyl group having from 2 to the indicated "n"
number of carbon atoms.
[0071] Unless the number of carbons is otherwise specified, "lower"
as in "lower aliphatic," "lower alkyl," "lower alkenyl," and "lower
alkylnyl", as used herein means that the moiety has at least one
(two for alkenyl and alkynyl) and equal or less than 6 carbon
atoms.
[0072] The terms "cycloalkyl", "alicyclic", "carbocyclic" and
equivalent expressions refer to a group comprising a saturated or
partially unsaturated carbocyclic ring in a single, spiro (sharing
one atom), or fused (sharing at least one bond) carbocyclic ring
system having from three to fifteen ring members. Examples of
cycloalkyl groups include, without limitation, cyclopropyl,
cyclobutyl, cyclopentyl, cyclopenten-1-yl, cyclopenten-2-yl,
cyclopenten-3-yl, cyclohexyl, cyclohexen-1-yl, cyclohexen-2-yl,
cyclohexen-3-yl, cycloheptyl, bicyclo[4,3,0]nonanyl, norbornyl, and
the like. The term cycloalkyl includes both unsubstituted
cycloalkyl groups and substituted cycloalkyl groups. The term
"C.sub.3-C.sub.ncycloalkyl", wherein n is an integer from 4 to 15,
refers to a cycloalkyl group having from 3 to the indicated "n"
number of carbon atoms in the ring structure. Unless the number of
carbons is otherwise specified, "lower cycloalkyl" groups as herein
used, have at least 3 and equal or less than 8 carbon atoms in
their ring structure.
[0073] The term "heterocycloalkyl" and equivalent expressions refer
to a group comprising a saturated or partially unsaturated
carbocyclic ring in a single, spiro (sharing one atom), or fused
(sharing at least one bond) carbocyclic ring system having from
three to fifteen ring members, including one to six heteroatoms
(e.g. N, O, S, P) or groups containing such heteroatoms (e.g. NH,
NR.sub.x (R.sub.x is alkyl, acyl, aryl, heteroaryl or cycloalkyl),
PO.sub.2, SO, SO.sub.2, and the like). Heterocycloalkyl groups may
be C-attached or heteroatom-attached (e.g. via a nitrogen atom)
where such is possible. Examples of heterocycloalkyl groups
include, without limitation, pyrrolidino, tetrahydrofuranyl,
tetrahydrodithienyl, tetrahydropyranyl, tetrahydrothiopyranyl,
piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl,
azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl,
thiepanyl, oxazepinyl, diazepinyl, thiazepinyl,
1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl,
2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl,
dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl,
dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,
3-azabicyclo[3,1,0]hexanyl, 3-azabicyclo[4,1,0]heptanyl,
3H-indolyl, quinolizinyl, and sugars, and the like. The term
heterocycloalkyl includes both unsubstituted heterocycloalkyl
groups and substituted heterocycloalkyl groups. The term
"C.sub.3-C.sub.nheterocycloalkyl", wherein n is an integer from 4
to 15, refers to a heterocycloalkyl group having from 3 to the
indicated "n" number of atoms in the ring structure, including at
least one hetero group or atom as defined above. Unless the number
of carbons is otherwise specified, "lower heterocycloalkyl" groups
as herein used, have at least 3 and equal or less than 8 carbon
atoms in their ring structure.
[0074] The terms "aryl" and "aryl ring" refer to aromatic groups
having "4n+2".pi.(pi) electrons, wherein n is an integer from 1 to
3, in a conjugated monocyclic or polycyclic system (fused or not)
and having six to fourteen ring atoms. A polycyclic ring system
includes at least one aromatic ring. Aryl may be directly attached,
or connected via a C.sub.1-C.sub.3alkyl group (also referred to as
arylalkyl or aralkyl). Examples of aryl groups include, without
limitation, phenyl, benzyl, phenetyl, 1-phenylethyl, tolyl,
naphthyl, biphenyl, terphenyl, indenyl, benzocyclooctenyl,
benzocycloheptenyl, azulenyl, acenaphthylenyl, fluorenyl,
phenanthernyl, anthracenyl, and the like. The term aryl includes
both unsubstituted aryl groups and substituted aryl groups. The
term "C.sub.6-C.sub.naryl", wherein n is an integer from 6 to 15,
refers to an aryl group having from 6 to the indicated "n" number
of atoms in the ring structure, including at least one hetero group
or atom as defined above.
[0075] The terms "heteroaryl" and "heteroaryl ring" refer to an
aromatic groups having "4n+2".pi.(pi) electrons, wherein n is an
integer from 1 to 3, in a conjugated monocyclic or polycyclic
system (fused or not) and having five to fourteen ring members,
including one to six heteroatoms (e.g. N, O, S) or groups
containing such heteroatoms (e.g. NH, NR.sub.x (R.sub.x is alkyl,
acyl, aryl, heteroaryl or cycloalkyl), SO, and the like). A
polycyclic ring system includes at least one heteroaromatic ring.
Heteroaryls may be directly attached, or connected via a
C.sub.1-C.sub.3alkyl group (also referred to as heteroarylalkyl or
heteroaralkyl). Heteroaryl groups may be C-attached or
heteroatom-attached (e.g. via a nitrogen atom), where such is
possible. Examples of heteroaryl groups include, without
limitation, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl,
tetrazolyl, furyl, thienyl; isooxazolyl, thiazolyl, oxazolyl,
isothiazolyl, pyrrollyl, quinolinyl, isoquinolinyl, indolyl,
isoindolyl, chromenyl, isochromenyl, benzimidazolyl, benzofuranyl,
cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,
pyrazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl,
thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,
benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl,
quinolizinyl, quinolonyl, isoquinolonyl, quinoxalinyl,
naphthyridinyl, furopyridinyl, carbazolyl, phenanthridinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxazinyl, dibenzofurnayl, and the like. The
term heteroaryl includes both unsubstituted heteroaryl groups and
substituted heteroaryl groups. The term
"C.sub.5-C.sub.nheteroaryl", wherein n is an integer from 6 to 15,
refers to an heteroaryl group having from 5 to the indicated "n"
number of atoms in the ring structure, including at least one
hetero group or atom as defined above.
[0076] The terms "heterocycle" or "heterocyclic" include
heterocycloalkyl and heteroaryl groups. Examples of heterocycles
include, without limitation, acridinyl, azocinyl, benzimidazolyl,
benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl,
benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazolinyl, carbazolyl,
4.alpha.H-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl,
decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,
dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl, imidazolinyl, imidazolyl, 1 H-indazolyl, indolenyl,
indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl,
isochromanyl, isoindazolyl, isoindolinyl, isoindolyl,
isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl,
morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,
1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,
1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl,
pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl,
pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,
pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,
1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl,
1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, xanthenyl, and
the like. The term heterocycle includes both unsubstituted
heterocyclic groups and substituted heterocyclic groups.
[0077] The term "amine" or "amino," as used herein, refers to an
unsubstituted or substituted moiety of the formula
--NR.sup.aR.sup.b, in which R.sup.a and R.sup.b are each
independently hydrogen, alkyl, aryl, or heterocyclyl, or R.sup.a
and R.sup.b, taken together with the nitrogen atom to which they
are attached, form a heterocyclic ring. The term amino includes
compounds or moieties in which a nitrogen atom is covalently bonded
to at least one carbon or heteroatom. Thus, the terms "alkylamino"
and "dialkylamino" as used herein means an amine group having
respectively one and at least two C.sub.1-C.sub.6alkyl groups
attached thereto. The term "acylamino" and "diarylamino" include
groups wherein the nitrogen is bound to at least one or two aryl
groups, respectively. The term "amide" or "aminocarbonyl" includes
compounds or moieties which contain a nitrogen atom which is bound
to the carbon of a carbonyl or a thiocarbonyl group. The term
acylamino refers to an amino group directly attached to an acyl
group as defined herein.
[0078] The term "nitro" means --NO.sub.2; the terms "halo" and
"halogen" refer to bromine, chlorine, fluorine or iodine
substituents; the term "thiol", "thio", or "mercapto" means SH; and
the term "hydroxyl" or "hydroxy" means --OH. The term "alkylthio"
refers to an alkyl group, having a sulfhydryl group attached
thereto. Suitable alkylthio groups include groups having 1 to about
12 carbon atoms, preferably from 1 to about 6 carbon atoms. The
term "alkylcarboxyl" as used herein means an alkyl group having a
carboxyl group attached thereto.
[0079] The term "alkoxy" or "lower alkoxy" as used herein means an
alkyl group having an oxygen atom attached thereto. Representative
alkoxy groups include groups having 1 to about 6 carbon atoms,
e.g., methoxy, ethoxy, propoxy, tert-butoxy and the like. Examples
of alkoxy groups include methoxy, ethoxy, isopropyloxy, propoxy,
butoxy, pentoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichioromethoxy, trichloromethoxy groups and the
like. The term alkoxy includes both unsubstituted or substituted
alkoxy groups., etc., as well as perhalogenated alkyloxy
groups.
[0080] The term "carbonyl" or "carboxy" includes compounds and
moieties which contain a carbon connected with a double bond to an
oxygen atom. Examples of moieties which contain a carbonyl include
aldehydes, ketones, carboxylic acids, amides, esters, anhydrides,
etc.
[0081] The term "acyl" refers to a carbonyl group that is attached
through its carbon atom to a hydrogen (i.e., formyl), an aliphatic
group (C.sub.1-C6alkyl, C.sub.1-C.sub.6alkenyl,
C.sub.1-C.sub.6alkynyl, e.g. acetyl), a cycloalkyl group
(C.sub.3-C.sub.3cycloalkyl), a heterocyclic group
(C.sub.3-C.sub.8heterocycloalkyl and C.sub.5-C.sub.6heteroaryl), an
aromatic group (C.sub.6aryl, e.g., benzoyl), and the like. Acyl
groups may be unsubstituted or substituted acyl groups (e.g.
salicyloyl).
[0082] It will be understood that "substitution" or "substituted
with" includes the implicit proviso that such substitution is in
accordance with the 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, etc. As used
herein, the term "substituted" is meant 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. The permissible substituents can
be one or more. The term "substituted", when in association with
any of the foregoing groups refers to a group substituted at one or
more position with substituents such as acyl, amino (including
simple amino, mono and dialkylamino, mono and diarylamino, and
alkylarylamino), acylamino (including carbamoyl, and ureido),
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
alkoxycarbonyl, carboxy, carboxylate, aminocarbonyl, mono and
dialkylaminocarbonyl, cyano, azido, halogen, hydroxyl, nitro,
trifluoromethyl, thio, alkylthio, arylthio, alkylthiocarbonyl,
thiocarboxylate, lower alkyl, lower alkenyl, lower alkynyl,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, lower alkoxy,
aryloxy, aryloxycarbonyloxy, benzyloxy, benzyl, sulfinyl,
alkylsulfinyl, sulfonyl, sulfate, sulfonate, sulfonamide,
phosphate, phosphonato, phosphinato, oxo, guanidine, imino, formyl
and the like. Any of the above substituents can be further
substituted if permissible, e.g. if the group contains an alkyl
group, an aryl group, or other.
[0083] The term "solvate" refers to a physical association of a
compound of this invention with one or more solvent molecules,
whether organic or inorganic. This physical association includes
hydrogen bonding. In certain instances, the solvate will be capable
of isolation, for example when one or more solvent molecules are
incorporated in the crystal lattice of the crystalline solid.
"Solvate" encompasses both solution-phase and isolable solvates.
Exemplary solvates include hydrates, ethanolates, methanolates,
hemiethanolates, and the like.
[0084] A "pharmaceutically acceptable salt" of a compound means a
salt of a compound that is pharmaceutically acceptable. Desirable
are salts of a compound that retain or improve the biological
effectiveness and properties of the free acids and bases of the
parent compound as defined herein or that takes advantage of an
intrinsically basic, acidic or charged functionality on the
molecule and that is not biologically or otherwise undesirable.
Example of pharmaceutically acceptable salts are also described,
for example, in Berge et al., "Pharmaceutical Salts", J. Pharm.
Sci. 66, 1-19 (1977). Such salts include:
[0085] (1) acid addition salts, formed on a basic or positively
charged functionality, by the addition of inorganic acids such as
hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric
acid, sulfamic acid, nitric acid, phosphoric acid, carbonate
forming agents, and the like; or formed with organic acids such as
acetic acid, propionic acid, lactic acid, oxalic, glycolic acid,
pivalic acid, t-butylacetic acid, .beta.-hydroxybutyric acid,
valeric acid, hexanoic acid, cyclopentanepropionic acid, pyruvic
acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric
acid, tartaric acid, citric acid, benzoic acid,
3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic
acid, 2-hydroxyethanesulfonic acid, cyclohexylaminosulfonic acid,
benzenesulfonic acid, sulfanilic acid, 4-chlorobenzenesulfonic
acid, 2-napthalenesulfonic acid, 4-toluenesulfonic acid,
camphorsulfonic acid, 3-phenyl propionic acid, lauryl sulphonic
acid, lauryl sulfuric acid, oleic acid, palmitic acid, stearic
acid, lauric acid, embonic (pamoic) acid, palmoic acid, pantothenic
acid, lactobionic acid, alginic acid, galactaric acid, galacturonic
acid, gluconic acid, glucoheptonic acid, glutamic acid, naphthoic
acid, hydroxynapthoic acid, salicylic acid, ascorbic acid, stearic
acid, muconic acid, and the like;
[0086] (2) base addition salts, formed when an acidic proton
present in the parent compound either is replaced by a metal ion,
including, an alkali metal ion (e.g. lithium, sodium, potassium),
an alkaline earth ion (e.g. magnesium, calcium, barium), or other
metal ions such as aluminum, zinc, iron and the like; or
coordinates with an organic base such as ammonia, ethylamine,
diethylamine, ethylenediamine, N,N'-dibenzylethylenediamine,
ethanolamine, diethanolamine, triethanolamine, tromethamine,
N-methylglucamine, piperazine, chloroprocain, procain, choline,
lysine and the like.
[0087] Pharmaceutically acceptable salts may be synthesized from
the parent agent that contains a basic or acidic moiety, by
conventional chemical methods. Generally, such salts are prepared
by reacting the free acid or base forms of these agents with a
stoichiometric amount of the appropriate base or acid in water or
in an organic solvent, or in a mixture of the two. Salts may be
prepared in situ, during the final isolation or purification of the
agent or by separately reacting a purified compound of the
invention in its free acid or base form with the desired
corresponding base or acid, and isolating the salt thus formed. The
term "pharmaceutically acceptable salts" also include zwitterionic
compounds containing a cationic group covalently bonded to an
anionic group, as they are "internal salts".
[0088] All acid, salt, base, and other ionic and non-ionic forms of
the compounds described are included as compounds of the invention.
For example, if a compound is shown as an acid herein, the salt
forms of the compound are also included. Likewise, if a compound is
shown as a salt, the acid and/or basic forms are also included.
[0089] "Abeta", "A.beta.", or ".beta.-amyloid", is defined as any
peptide resulting from beta-secretase mediated cleavage of Beta
Amyloid Precursor Protein (APP), including for examples peptides of
37, 38, 39, 40, 41, 42, and 43 amino acids, and extending from the
beta-secretase cleavage site to amino acids 37, 38, 39, 40, 41, 42,
or 43. It also includes It also includes N-terminal truncated
species of above peptides, such as the pyroglutamic forms pE3-40,
pE3-42, pE3-43, pE11-42, pE11-43 and the like. For convenience of
nomenclature, "A.beta..sub.1-42", may be referred to herein as
"A.beta.(1-42)" or simply as "A.beta..sub.42" (and likewise for any
other amyloid peptides discussed herein). As used herein, the terms
"Abeta", "A.beta.", ".beta.-amyloid", "amyloid-.beta." are
synonymous referring collectively to truncated and non-truncated
peptide species of the sequence between .beta.- and
.gamma.-cleavage sites of APP.
[0090] The term "amyloid-.beta. disease" or "amyloid-.beta. related
disease" may be used for mild cognitive impairment; vascular
dementia; early Alzheimer's disease; Alzheimer's disease, including
sporadic (non-hereditary) Alzheimer's disease and familial
(hereditary) Alzheimer's disease; age-related cognitive decline;
cerebral amyloid angiopathy ("CAA"); hereditary cerebral
hemorrhage; senile dementia; Down's syndrome; inclusion body
myositis ("IBM"); or age-related macular degeneration ("ARMD"),
Mild cognitive impairment ("MCI"), Cerebral amyloid angiopathy
("CAA"), age-related macular degeneration (ARMD).
[0091] "AUC" is the area under a curve representing the
concentration of a compound in a biological sample of a subject as
a function of time following administration of the compound to the
subject. Examples of biological samples include biological fluids
such as plasma and blood, or organ homogenates such as brain or
liver homogenates. The AUC can be determined by measuring the
concentration of a compound in a biological sample such as the
plasma, blood or brain homogenate using methods such as liquid
chromatography-tandem mass spectrometry (LC/MS/MS), at various time
intervals, and calculating the area under the
concentration-versus-time curve. Suitable methods for calculating
the AUC from a drug concentration-versus-time curve are well known
in the art. As relevant to the disclosure here, an AUC for SAPS can
be determined by measuring the concentration of SAPS in the plasma,
blood or brain homogenate of a subject following oral
administration of a compound of Formulae (I), (I-A), (I-C), (I-D),
(I-E), (I-P), (I-P2), (II), (III), (IV), (V), (VI), (VII), (VIII),
(IX), (X), (XI), (XII) or (XII-A), to the subject. Unless noted
otherwise herein; AUC means AUC.sub.0-.infin., as further defined
in Example 4.
[0092] "Bioavailability" refers to the rate and amount of a drug
that reaches the systemic circulation of a subject following
administration of the drug or prodrug thereof to the patient and
can be determined by evaluating, for example, the plasma or blood
concentration-versus-time profile for the drug. Parameters useful
in characterizing a plasma or blood concentration-versus-time curve
include the area under the curve (AUC), the time to peak
concentration (T.sub.max), and the maximum drug concentration
(C.sub.max). Bioavailability is often expressed as F(%) referring
to the ratio in percentage of the AUC of the compound for a
specific mode of administration (e.g. orally) over AUC of the
compound after an IV administration.
[0093] "Bioequivalence" refers to equivalence of the rate and
extent of absorption of a drug after administration of equal doses
of the drug or prodrug to a patient. As used herein, two plasma or
blood concentration profiles are bioequivalent if the 90%
confidence interval for the ratio of the mean response of the two
profiles is within the limits of 0.8 and 1.25. The mean response
includes at least one of the characteristic parameters of a profile
such as C.sub.max, T.sub.max, and AUC.
[0094] "C.sub.max" is the maximum concentration of a drug in the
biological sample of a subject following administration of a dose
of the drug or prodrug to the subject. "T.sub.max" is the time to
the maximum concentration (C.sub.max) of a drug in the biological
sample of a subject following administration of a dose of the drug
or prodrug to the subject.
[0095] As used herein the term "effective amount" refers to the
amount or dose of the compound, upon single or multiple dose
administration to the patient, which provides the desired effect in
the patient under diagnosis or treatment. An effective amount can
be readily determined by the attending diagnostician, as one
skilled in the art, by the use of known techniques and by observing
results obtained under analogous circumstances. In determining the
effective amount or dose of compound administered, a number of
factors are considered by the attending diagnostician, including,
but not limited to: the size, age, and general health of the
subject; the specific disease involved; the degree of or
involvement or the severity of the disease; the response of the
individual subject; the particular compound administered; the mode
of administration; the bioavailability characteristics of the
preparation administered; the dose regimen selected; the use of
concomitant medication; and other relevant circumstances.
[0096] As used herein the term "therapeutic bio-distribution of
3APS" refers to one or more pharmacokinetic parameters of SAPS
which affect 3APS therapeutic activity. Examples of such
pharmacokinetic (PK) parameters include but are not limited to:
bioavailability of 3APS, AUC of 3APS, brain levels of 3APS, CSF
levels of 3APS, C.sub.max of 3APS , T.sub.max of 3APS, and/or
bio-absorption of 3APS, etc.
[0097] As used herein the terms "increased (or like terms, e.g.,
increasing, increase in, etc.) therapeutic effectiveness of 3APS"
and "enhanced (or like terms, e.g., enhancing, enhancement, etc.)
therapeutic effectiveness of 3APS" refer to an increased
effectiveness of 3APS as measured, e.g., by one or more parameters
listed under "therapeutic bio-distribution of 3APS" above, e.g., by
5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 125%,
etc., or even more, e.g., 2, or 4 fold, or even more when
administered to a subject, e.g., animal or human, which increase is
with respect to the same equivalent molar dose of 3APS administered
orally in water solution. Preferably such % increases are achieved
also with respect to 3APS administered orally in the formulation of
Table 3 of U.S. Ser. No. 11/103,656, filed on Apr. 12, 2005.
Effectiveness can also be as measured, for example, by effect on
characteristics of a disease such as Alzheimer's disease, e.g., by
the reduction of plaques or A.beta. load in the brain, or by an
improvement in selected manifestations of the disease, e.g., memory
loss, cognition, reasoning, judgment, orientation, etc. See U.S.
Ser. No. 11/103,656, filed on Apr. 12, 2005, for details on how to
measure effects on characteristics of such diseases.
[0098] The term "lessening metabolism of 3APS" (or related terms
such as reduction, less, lowering, reducing, lowered, etc) refers
to decreasing the degree or amount of first-pass metabolism in the
GI tract or liver of 3APS (by administering it to a subject
non-orally or in particular oral formulations or in the form of a
prodrug) by e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,
95%, 99%, or even 100%, which decrease is with respect to the
degree or amount of metabolism of SAPS that occurs when the same
equivalent molar dose of SAPS is administered orally in water
solution. Preferably such % decreases are achieved also with
respect to SAPS administered orally in the formulation of Table 3
of U.S. Ser. No. 11/103,656, filed on Apr. 12, 2005.
[0099] The term "reduction of side effects of 3APS" refers to
decreasing the amount of or severity of one or more side effects of
3APS by, e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,
95%, 99%, or 99.9%, or even 100%, which decrease is with respect to
the amount of or severity of a side effect of 3APS that is
exhibited when the same equivalent molar dose of 3APS is
administered orally in water solution. Preferably such % decreases
are achieved also with respect to 3APS administered orally in the
formulations of Table 3 of U.S. Ser. No. 11/103,656, filed on Apr.
12, 2005.
[0100] More generally, the terms lessening etc., increasing etc.,
refer in context herein to the percentage changes, e.g., by 5%,
10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 125%, etc.,
or even more, e.g., 2, or 4 fold, or even more.
[0101] All of the pharmacokinetic data in U.S. Ser. No. 11/103,656,
filed on Apr. 12, 2005, are incorporated herein by reference,
including the data for example 1 and of Table 3, for example, for
forming a comparative basis for the effects achieved by the present
inventions.
[0102] When referring to "3APS" being produced (e.g., released from
a formulation or prodrug), all forms of 3APS are included, e.g.,
solvates thereof, ionically dissociated forms thereof, charged
forms thereof, etc.
[0103] "Pharmaceutically acceptable" refers to drugs, medicaments,
inert ingredients etc., which the term describes, suitable for use
in contact with the tissues of humans and lower animals without
undue toxicity, incompatibility, instability, irritation, allergic
response, and the like, commensurate with a reasonable benefit/risk
ratio. It preferably refers to a compound or composition that is
approved or approvable by a regulatory agency of the Federal or
state government or listed in the U.S. Pharmacopoeia or other
generally recognized pharmacopoeia for use in animals and more
particularly in humans.
[0104] "Pharmaceutically acceptable vehicle" refers to a diluent,
adjuvant, excipient, or carrier with which a compound is
administered.
[0105] "Pharmaceutical composition" refers to at least one compound
and at least one pharmaceutically acceptable vehicle, with which
the compound is administered to a patient.
[0106] "Preventing" or "prevention" is intended to refer at least
the reduction of likelihood of the risk of (or susceptibility to)
acquiring a disease or disorder (i.e., causing at least one of the
clinical symptoms of the disease not to develop in a patient that
may be exposed to or predisposed to the disease but does not yet
experience or display symptoms of the disease).
[0107] "Treating" or "treatment" of any disease or disorder refers,
in some embodiments, to ameliorating at least one disease or
disorder (i.e., arresting or reducing the development of the
disease or at least one of the clinical symptoms thereof). In
certain embodiments "treating" or "treatment" refers to
ameliorating at least one physical parameter, which may or may not
be discernible by the patient. In certain embodiments, "treating"
or "treatment" refers to inhibiting the disease or disorder, either
physically, (e.g., stabilization of a discernible symptom),
physiologically, (e.g., stabilization of a physical parameter), or
both. In certain embodiments, "treating" or "treatment" refers to
delaying the onset of the disease or disorder. The term "treating"
refers to any indicia of success in the treatment or amelioration
of an injury, pathology or condition, including any objective or
subjective parameter such as abatement; remission; diminishing of
symptoms or making the injury, pathology or condition more
tolerable to the subject; slowing in the rate of degeneration or
decline; making the final point of degeneration less debilitating;
improving a subject's physical or mental well-being; or, in some
situations, preventing the onset of dementia. The treatment or
amelioration of symptoms can be based on objective or subjective
parameters; including the results of a physical examination, a
psychiatric evaluation, or a cognition test such as CDR, MMSE, DAD,
ADAS-Cog, or another test known in the art. For example, the
methods of the invention successfully treat a subject's dementia by
slowing the rate of or lessening the extent of cognitive
decline.
[0108] "Therapeutically effective amount" means the amount of
compound that, when administered to a patient for treating or
preventing a disease, is sufficient to effect such treatment or
prevention of the disease. The "therapeutically effective amount"
will vary depending on the compound, the disease and its severity,
and the age, weight, etc., of the patient having the disease to be
treated or prevented.
[0109] Reference will now be made in detail to certain embodiments
of compounds and, methods. The disclosed embodiments are not
intended to be limiting of the invention.
[0110] In a further aspect the invention includes the
administration of SAPS that is not via a transdermal patch, or not
by topical administration in a composition, e.g., lotions, creams,
solutions, gels or solids or not by subcutaneous, intravenous or
intraperitoneal injection, or not intraspinally, or
intracerebrally.
II. Compounds of the Invention
[0111] The present invention relates to methods, compounds and
compositions for delivering in a subject, preferably a human
subject, 3-amino-1-propanesulfonic acid, or salts thereof, also
referred herein as 3APS. The invention encompasses compounds that
will yield or generate 3APS, either in vitro or in viva
[0112] In a preferred embodiment, compounds of the invention
include prodrugs that will yield or generate 3APS once administered
in a human. Without wishing to be bound by theory, in some aspects
the prodrugs according to invention comprise a "pharmacokinetic
modulating moiety," e.g., B, below, covalently but dissociably
linked to 3APS that, e.g., by a linkage, L, below, which will be
cleaved once in the blood, plasma or other specific tissue (e.g.
brain), thereby releasing 3APS.
[0113] Thus, in one aspect, the invention relates to a compound of
the Formula I:
B-L-A (I)
as well as pharmaceutically acceptable salts, metabolites, and
solvates thereof, where: B is a pharmacokinetic modulating moiety,
which is optionally also bonded to A directly or indirectly thought
a further linking group L; A is 3-amino-1-propanesulfonic acid
moiety (i.e, 3APS bound to L-B); and L is a cleavable linkage for
covalently and dissociably coupling B to A (preferably and
typically via the NH.sub.2 group), or is absent, whereby L can be a
direct bond or additional chemical structure providing a cleavable
linkage.
[0114] Suitable pharmacokinetic modulating moieties (e.g. B)
moieties include amino acid or peptide moieties, carbamate
moieties, non-amino-acid amide moieties, carbohydrate-derived
moieties and analogs such as inositol-derived moieties, N-hydroxy
and derivatives thereof (e.g., where the H in OH is replaced by an
OH protecting group). B can also comprise a cyclic double protected
3APS molecule and precursors (e.g., where a moiety connects
NH.sub.2 and SO.sub.3H of 3APS, e.g., sulfinic acids, thiols,
sulfides, disulfides, etc.), and combinations thereof. More
generally B moieties include N-protecting groups. B can also be the
molecule 3APS itself (see gemini dimers).
[0115] Suitable linkages L will be any which are cleaved as
described herein, e.g., by enzymes mentioned herein or others in
blood, plasma and or brain cells, in vitro or in vivo. Linkages
will generally comprise a bond which is known to be so cleavable
such as but not limited to, a peptide, amide, ester, sulfide,
disulfide, carboxamate, urea, --N--O--, etc. bond, and others as
demonstrated for example in the structures disclosed herein, all of
which are in general applicable as linkages, L, in compounds in
general. Actual cleavability of the linker can be assessed in vitro
and/or in vivo by using hydrolytic-, enzymatic- (e.g. peptidase,
esterase) or metabolic-based tests and assays well known in the
art. International PCT application WO 91/14434, published
applications US 2005/0096317, US 2006/0046967 and provisional
application U.S. Ser. No. 60/xxx,xxx filed concurrently herewith,
are all incorporated herein by reference since they describe a
variety of linkers that may be useful according to the present
invention.
[0116] In another aspect, the invention relates to Formula I-A (and
salts, esters and solvates thereof)
##STR00003##
wherein,
[0117] R.sup.x and R.sup.y are independently selected from hydrogen
and a protecting group, wherein R.sup.x and R.sup.y are not both
hydrogen; and
[0118] L.sup.1 and L.sup.2 are each a cleavable linkage; wherein
when R.sup.x is H, L.sup.1 is absent, and when R.sup.y is H, then
L.sup.2 is absent.
[0119] The term "protecting group" refers to a group inhibiting and
reducing metabolism of the amino group of 3APS. Examples of
protecting groups include, without limitation, an amino acid
residue, a carbamate, a non-amino acid amide, a
carbohydrate-derived residue, a N-hydroxy-derived residue, a cyclic
double protecting group, and the like.
[0120] According to preferred embodiments, the compounds of the
invention exhibit numerous advantageous properties. In one
embodiment, the compound is a prodrug which bypasses first-pass
metabolism by the liver and/or the digestive tract (e.g. gut,
stomach, or intestine) that is associated with administration of
3APS, per se, thereby increasing bio-distribution and/or
bioavailability of 3APS as compared to an administration of a molar
equivalent of 3APS. Bypassing hepatic fist-pass metabolism
modifies, improves or increases pharmacokinetic parameters of 3APS
such as the AUC, the C.sub.max and/or T.sub.max of 3APS. In one
embodiment, the compound is a prodrug which exhibits an increased
absorption by the gastrointestinal tract, compared to the
administration a molar equivalent of 3APS per se. In one
embodiment, the compound is a prodrug which provides a slow release
of 3APS over time. In another embodiment, the compound is a prodrug
which increases brain levels of 3APS when compared to the
administration a molar equivalent of 3APS per se. In another
embodiment, the compound is a prodrug which lessens common side
effects associated with the administration of 3APS per se. For
instance, in a preferred embodiment, the prodrug exhibits a better
gastrointestinal tolerability than 3APS.
[0121] In preferred embodiments the compounds and/or compositions
of the invention achieve one or more of the following benefits: (1)
reducing the molar dose of 3APS administered to a patient (e.g. due
to a improved absorption when compared to 3APS or due to a
reduction in the first-pass metabolism of 3APS); (2) avoiding
common side effects such as gastrointestinal irritation associated
with an oral administration of 3APS; (3) improving penetration of
3APS across the BBB; (4) reducing the side effects associated with
3APS (e.g. by lessening gastrointestinal problems or by increasing
the relative amount of 3APS reaching the brain; (5) increasing
concentration or levels of 3APS in desired tissues or fluids (e.g.
brain, CSF). Other benefits will be apparent to those skilled in
the art.
[0122] The invention pertains to both salt forms and acid/base
forms of the compounds of the invention. For example, the invention
pertains not only to the particular salt forms of compounds shown
herein as salts, but also the invention includes other
pharmaceutically acceptable salts, and the acid and/or base form of
the compound. The invention also pertains to salt forms of
compounds shown herein. Compounds of the invention are also shown
in Table 1, Table 2, Table 3, Table 4 and Table 4B below.
[0123] The compounds of the present invention may exhibit
polymorphism. Polymorphs of compounds according to this invention
may be prepared by crystallization under different conditions. For
example, using different solvents or different solvent mixtures for
recrystallization; crystallization at different temperatures;
various modes of cooling ranging from very fast to very slow
cooling during crystallization. Polymorphs may also be obtained by
heating or melting a prodrug followed by gradual or fast cooling.
The presence of polymorphs may be determined by solid probe NMR
spectroscopy, IR spectroscopy, differential scanning calorimetry,
powder X-ray diffraction or other such techniques.
[0124] The compounds of the present invention may also exist in the
form of a solvate, for example, hydrate, ethanolate, n-proponalate,
iso-propanolate, 1-butanolate, 2-butanolate and solvates of other
physiologically acceptable solvents, such as the Class 3 solvents
described in the International Conference on Harmonization (ICH),
Guidance for Industry, Q3C Impurities: Residual Solvents (1997).
The present invention includes each solvate and mixtures
thereof.
[0125] The amino acid or peptidic moiety, the carbamate moiety, the
non-amino acid amide moiety, the carbohydrate-derived moiety and
analogs such as inositol-derived, the N-hydroxy moiety and
derivatives, or any other pharmacokinetic modulating moiety of the
prodrugs, including cyclic double protected 3APS and precursors
(e.g. sulfinic acids, thiol, sulfide, disulfide, etc), and
combinations thereof, according to the invention may be cleaved
prior to absorption by the gastrointestinal tract (e.g., within the
stomach or intestinal lumen) and/or after absorption by the
gastrointestinal tract (e.g., in intestinal tissue, blood, liver,
or other suitable tissue of a mammal). In certain embodiments, 3APS
remains covalently attached to the pharmacokinetic modulating
moiety during transit across the intestinal mucosal barrier to
provide protection from presystemic metabolism. In certain
embodiments, pharmacokinetic modulating moieties according to the
invention are essentially not metabolized to the corresponding 3APS
within cells of the intestine or liver (e.g. enterocytes,
hepatocytes), but generates the parent 3APS molecule once within
the systemic circulation. In certain embodiments, at least some of
the prodrug administered generates the corresponding SAPS only once
in the brain, i.e. after it has passed the blood brain barrier
(BBB). Cleavage of the pharmacokinetic modulating moiety of
prodrugs according to the invention after absorption by the
gastrointestinal tract may allow these prodrugs to be absorbed into
the systemic circulation either by active transport, passive
diffusion, or by a combination of both active and passive
processes. Accordingly, in certain embodiments, a pharmaceutical
composition, formulation, or dosage form of the present invention
is capable of maintaining a therapeutically effective concentration
of SAPS in the plasma or blood of a patient for a time period of at
least about 1 hour, for at least 2 hours, for at least 3 hours, 4
hours, for at least about 8 hours, for a period of at least about
12 hours, at least about 16 hours, at least about 20 hours, and in
certain embodiments for at least about 24 hours after the
pharmaceutical composition, formulation, or dosage form comprising
a corresponding compound according to the invention and a
pharmaceutically acceptable vehicle is orally administered to the
patient. In certain embodiments, a pharmaceutical composition,
formulation, or dosage form of the present invention is capable of
improving the T.sub.max of SAPS by at least 2 fold, or by at least
3, 4, 5, 6, 7, 8, 9 or 10 fold or more.
[0126] The pharmacokinetic modulating moiety of certain of the
compounds according to the invention may be cleaved either
chemically and/or enzymatically. One or more enzymes present in the
stomach, intestinal lumen, intestinal tissue, blood, liver, brain,
or any other suitable tissue of a mammal may enzymatically cleave
the amino acid or peptidic moiety of the compound. If the
pharmacokinetic modulating moiety is cleaved after absorption by
the gastrointestinal tract, certain of the compounds according to
the invention may have the opportunity to be absorbed into the
systemic circulation from the large intestine. In certain
embodiments, the pharmacokinetic modulating moiety is cleaved after
absorption by the gastrointestinal tract or after crossing the
BBB.
[0127] Although theory of operation is discussed herein, for
specific compound structures, including all generic structural
formulas and specific names and formulas of compounds, the
invention is not limited by any such theories unless specifically
stated otherwise. Thus, all uses of all novel compounds are
encompassed by the invention, irrespective of mechanism or theory
of operation.
Il-A. Amino Acid Prodrugs
[0128] In a preferred embodiment, the compounds of the invention
are amino acids prodrugs that will yield or generate 3APS once
administered in a human. Preferred prodrugs are composed of an
amino acid residue linked to the amine group of 3APS via an amide
bond. The amino acid residue may be cleaved in vivo by enzymes such
as peptidases, or by any other mechanisms, to liberate the amine
group of 3APS.
[0129] More particularly, an aspect of the invention relates to a
compound of Formula (II), and to pharmaceutically acceptable salts,
esters or solvates thereof:
##STR00004##
wherein AA is a natural or unnatural amino acid residue or a
peptide comprising 2, 3 or more natural or unnatural amino acid
residues.
[0130] Other aspects of the invention relate to compounds of
Formula (III) and to a pharmaceutically acceptable salt or solvate
thereof:
##STR00005##
wherein:
[0131] aa.sup.1 is a natural or unnatural amino acid residue;
[0132] aa.sup.2 and aa.sup.3 are each independently a natural or
unnatural amino acid residue or absent.
[0133] Further aspects of the invention relate to compounds of
Formula (IV) and to pharmaceutically acceptable salts, esters or
solvates thereof:
##STR00006##
wherein:
[0134] aa.sup.1 is a natural or unnatural amino acid residue;
[0135] aa.sup.2 is a natural or unnatural amino acid residue, or is
absent.
[0136] Yet further aspects of the invention relate to compounds of
Formula (V), and to pharmaceutically acceptable salts, esters or
solvates thereof:
##STR00007##
wherein aa.sup.1 is a natural or unnatural amino acid residue.
[0137] The invention further relates to compounds of Formula (V-A),
and to pharmaceutically acceptable salts, esters or solvates
thereof:
##STR00008##
wherein aa.sup.x is an amino acid residue selected from valine,
proline, lysine, leucine, methionine, D-methionine, serine,
alanine, D-alanine, glycine, isoleucine, histidine, aminoisobutyric
acid, phenylglycine, tryptophan, tyrosine, O-benzylserine,
O-benzylglutamine, and .gamma.-aminobutyric acid.
[0138] In preferred embodiments aa.sup.x is an amino acid residue
selected from valine, lysine, methionine, serine, and
O-benzylserine, or a pharmaceutically acceptable salt or solvate
thereof.
[0139] In one embodiment, the amino acid residue is coupled via an
acid end (C-coupled). In an embodiment, the amino acid residue is a
natural amino acid residue, or a salt or ester thereof. In another
embodiment, the amino acid residue is an unnatural amino acid
residue, or a salt or ester thereof. In yet another embodiment, the
amino acid residue is not a phenylalanine, e.g., in the case where
a single amino acid is attached to the N atom, but also in any
other case. In a further embodiment, natural or unnatural amino
acid residues in Formula II, Formula III, Formula IV, Formula V, or
Formula V-A are optionally represented by Formula (VI):
##STR00009##
wherein:
[0140] R.sup.1 and R.sup.2 are each independently selected from the
group consisting of H and a substituted or unsubstituted group
selected from C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl, C.sub.3
-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl,
C.sub.5-C.sub.15heteroaryl, NH(C.sub.1-C.sub.6alkyl),
N(C.sub.1-C.sub.6alkyl).sub.2, and C(O)(C.sub.1-C.sub.6alkyl); or
R.sup.1 and R.sup.2 are taken together with the adjacent carbon
atom to form a substituted or unsubstituted
C.sub.3-C.sub.12heterocycloalkyl;
[0141] R.sup.3 is selected from the group consisting of H and a
substituted or unsubstituted group selected from
C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl,
C.sub.6-C.sub.15heteroaryl, C(O)(C.sub.1-C.sub.6alkyl), and
C(O)(C.sub.6-C.sub.10aryl); or R.sup.3 is a bond between two amino
acid residues, when at least two amino acid residues are
present;
[0142] R.sup.4 is selected from the group consisting of H and a
substituted or unsubstituted group selected from
C.sub.1-C.sub.6alkyl, C.sub.2-C6alkenyl, C.sub.2-C.sub.6alkynyl; or
R.sup.1 and R.sup.4 are taken together with the adjacent carbon and
nitrogen atoms to form a C.sub.3-C.sub.10heterocycloalkyl; and n is
1, 2 or 3, or a higher number.
[0143] In one embodiment, the compound of the invention comprises
an amino acid residue of Formula VI, wherein R.sup.2 is H and all
other groups are as previously disclosed. In another embodiment,
the compound of the invention comprises an amino acid residue of
Formula VI, wherein R.sup.2 and R.sup.3 are each H and all other
groups are as previously disclosed. In another embodiment, the
compound of the invention comprises an amino acid residue of
Formula VI, wherein when R.sup.2 and R.sup.3 are each H, then
R.sup.1 is not an aryl-substituted C.sub.1alkyl. In another
embodiment, the compound of the invention comprises an amino acid
residue of Formula VI, wherein when R.sup.2 and R.sup.3 are each H,
then R.sup.1 is not a -CH.sub.2aryl group. In another embodiment,
the compound of the invention comprises an amino acid residue of
Formula VI, wherein when R.sup.2 is H and R.sup.3 is H or a bond,
then R.sup.1 is not a --CH.sub.2phenyl group. In another
embodiment, the invention provides compounds of Formula V, provided
that aa.sup.1 is not a phenylalanine. In another embodiment, the
invention provides compounds of Formula IV, provided that aa.sup.1
and aa.sup.2 are not both D-phenylalanine. In another embodiment,
the invention provides compounds of Formula IV, provided that
aa.sup.1 and aa.sup.2 are not both L-phenylalanine. In another
embodiment, the invention provides compounds of Formula IV,
provided that when one of aa.sup.1 and aa.sup.2 is D-phenylalanine,
then the other is not D-phenylalanine or D-tyrosine. In yet another
embodiment, the invention provides compounds of Formula IV,
provided that when one of aa.sup.1 and aa.sup.2 is L-phenylalanine,
then the other is not D-phenylalanine or L or D-tyrosine.
TABLE-US-00001 TABLE 1 Exemplary amino acid prodrugs according to
the invention ID Structure A1 ##STR00010## A2 ##STR00011## A3
##STR00012## A4 ##STR00013## A5 ##STR00014## A6 ##STR00015## A7
##STR00016## A8 ##STR00017## A9 ##STR00018## A10 ##STR00019## A11
##STR00020## A12 ##STR00021## A13 ##STR00022## A14 ##STR00023## A15
##STR00024## A16 ##STR00025## A17 ##STR00026## A18 ##STR00027## A19
##STR00028## A20 ##STR00029## A21 ##STR00030## A22 ##STR00031## A23
##STR00032## A24 ##STR00033## A25 ##STR00034## A26 ##STR00035## A27
##STR00036## A28 ##STR00037## A29 ##STR00038## A30 ##STR00039## A31
##STR00040## A32 ##STR00041## A33 ##STR00042## A34 ##STR00043## A35
##STR00044##
[0144] Preferred amino acid prodrugs according to the invention are
Compounds A2, A4, A6, A7 and A18 (as described above), and
pharmaceutically acceptable salts and solvates thereof.
II-B. Carbamate, Non-Amino Acid Amide and Related Prodrugs
[0145] Certain aspects of the invention relate to a compound of
Formula (VII), and to pharmaceutically acceptable salts, esters or
solvates thereof:
##STR00045##
wherein,
[0146] R.sup.5 is a substituted or unsubstituted group selected
from C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl,
C.sub.6-C.sub.15heteroaryl, NH(C.sub.1-C.sub.6alkyl),
N(C.sub.1-C.sub.6alkyl).sub.2, and C(O)(C.sub.1-C.sub.6alkyl);
[0147] R.sup.6 is a hydrogen or a substituted or unsubstituted
group selected from C(O)NH.sub.2, C(O)NH(C.sub.1-C.sub.6alkyl),
C(O)N(C.sub.1-C.sub.6alkyl).sub.2, and C(O)(C.sub.1-C.sub.6alkyl);
or R.sup.5 and R.sup.6 are taken together with the adjacent carbon
atom to form a substituted or unsubstituted
C.sub.3-C.sub.12heterocycloalkyl;
[0148] M is selected from the group consisting of oxygen, sulfur,
and nitrogen (NH or N(C.sub.1-C.sub.6alkyl)) or is absent.
[0149] The invention pertains to both salt forms and acid/base
forms of the compounds of the invention. For example, the invention
pertains not only to the particular salt forms of compounds shown
herein as salts, but also the invention includes other
pharmaceutically acceptable salts, and the acid and/or base form of
the compound. The invention also pertains to salt forms of
compounds shown herein.
[0150] In one embodiment, the invention provides compounds of
Formula VII, wherein when M is absent and R.sup.6 is H, then
R.sup.5 is other than 1-(4-isobutylphenyl)ethyl. In another
embodiment, the invention provides compounds of Formula VII,
wherein when R.sup.6 is H and M is NH or absent, then R.sup.5 is
other than 1-(4-isobutylphenyl)ethyl. In another embodiment, the
invention provides compounds of Formula VII, wherein when R.sup.6
is H and M is NH, then R.sup.5 is other than benzyl,
diphenylmethyl, hexyl, dodecyl, adamantyl, and t-butyl. In another
embodiment, the invention provides compounds of Formula VII,
wherein when R.sup.6 is H and M is NH, then R.sup.5 is other than
hydrogen, 1,4-dihydro-5,6-dimethyl-4-oxo-2-pyrimidinyl, and
5-ethyloxycarbonyl-1-penthyl. In another embodiment, the invention
provides compounds of Formula VII, wherein when M is NH and R.sup.5
and R.sup.6 are taken together with the adjacent carbon atom to
form a substituted or unsubstituted C.sub.3-12heterocycloalkyl,
then the heterocycloalkyl is other than benzimidazol-2-one,
tetrahydro-2,4,6-trioxo-1,3,5-triazine, 2,4-dioxo-1-imidazolidine,
2,4-dioxo-(di or tetrahydro)-benzo[g]pteridine,
4,10-dihydro-10-methyl-2,4-dioxopyrimido[4,5b]quinoline,
2-oxo-1-imidazolidinyl, and 3,4-dihydro-2,4-dioxo-1(2H)pyrimidine.
In another embodiment, the invention provides compounds of Formula
VII, wherein when M is NH, then R.sup.5 and R.sup.6 are not taken
together with the adjacent carbon atom to form a substituted or
unsubstituted C.sub.3-12heterocycloalkyl. In another embodiment,
the invention provides compounds of Formula VII, wherein when
R.sup.6 is H and M is O, then R.sup.5 is other than t-butyl and
benzyl. In another embodiment, the invention provides compounds of
Formula VII, wherein when R.sup.6 is H and M is O, then R.sup.5 is
other than i-butyl and 9H-fluoren-9-ylmethyl. In another
embodiment, the invention provides compounds of Formula VII,
wherein when R.sup.6 is H and M is absent, then R.sup.5 is other
than benzyl, phenyl, 3-pyridinyl, 3-N-methylpyridinium, methyl,
trifluoromethyl, pentafluoroethyl, pentafluorophenyl, and t-butyl.
In another embodiment, the invention provides compounds of Formula
VII, wherein when R.sup.6 is H and M is absent, then R.sup.5 is
other than n-butyl, i-butyl, n-propyl, i-propyl, vinyl, 2-propenyl,
2-(1-decenyl), 2-(1-dodecenyl), 1-(8-undecenyl), octyl, decyl,
undecyl, tridecyl, pentadecyl, heptadecyl,
4-(N-oxy-2,2,6,6-tetramethylpiperidinyl),
5-(1,3-dihydro-1,3-dioxo-2-benzofuranyl), 4-nitrophenyl, and
3-phenoxyphenyl.
a) Carbamate Prodrugs
[0151] In some preferred embodiments, the compounds of the
invention are carbamate prodrugs that will yield or generate 3APS
once administered in a human. Preferred prodrugs comprise an
oxycarbonyl residue (--OC(O)--) linked to the amine group of 3APS
via a carbamate bond (--OC(O)--NH--). The amine residue may be
cleaved in vivo by enzymes or by any other mechanisms, including
hydrolysis, to liberate the amine group of 3APS. In a preferred
embodiment, the compounds of the invention are carbamate prodrugs
that will yield or generate 3APS once administered in a human.
[0152] More particularly, certain aspects of the invention relate
to a compound of Formula (VIII), and to pharmaceutically acceptable
salts, esters or solvates thereof:
##STR00046##
wherein,
[0153] R.sup.7 is a substituted or unsubstituted group selected
from C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.5-C.sub.15aryl,
C.sub.5-C.sub.15heteroaryl, C.sub.7-C.sub.12arylalkyl,
C.sub.7-C.sub.12heteroarylalkyl, and combinations thereof.
[0154] In one embodiment, the definition of R.sup.7 is a
substituted or unsubstituted 1-(alkylcarboxy)alkyl group. In
another embodiment, R.sup.7 is a substituted or unsubstituted
benzyl group. In another embodiment, R.sup.7 is a substituted or
unsubstituted heterocycloalkylmethylene group. In another
embodiment, the invention provides compounds of Formula VIII,
provided that R.sup.7 is other than t-butyl or benzyl. In another
embodiment, the invention provides compounds of Formula VIII,
provided that R.sup.7 is other than i-butyl or
9H-fluoren-9-ylmethyl.
[0155] The invention pertains to both salt forms and acid/base
forms of the compounds of the invention. For example, the invention
pertains not only to the particular salt forms of compounds shown
herein as salts, but also the invention includes other
pharmaceutically acceptable salts, and the acid and/or base form of
the compound. The invention also pertains to salt forms of
compounds shown herein. Compounds of the invention are also shown
in Table 2 below.
TABLE-US-00002 TABLE 2 Exemplary carbamate prodrugs according to
the invention ID Structure C1 ##STR00047## C2 ##STR00048## C3
##STR00049## C4 ##STR00050## C5 ##STR00051## C6 ##STR00052## C7
##STR00053## C8 ##STR00054## C9 ##STR00055## C10 ##STR00056## C11
##STR00057## C12 ##STR00058## C13 ##STR00059## C14 ##STR00060## C15
##STR00061## C16 ##STR00062## C17 ##STR00063## C18 ##STR00064## C19
##STR00065## C20 ##STR00066## C21 ##STR00067## C22 ##STR00068## C23
##STR00069## C24 ##STR00070## C25 ##STR00071## C26 ##STR00072##
b) Non-Amino Acid Amide Prodrugs
[0156] In some preferred embodiments, the compounds of the
invention are non-amino acid amide prodrugs that will yield or
generate SAPS once administered to a human. Preferred prodrugs
comprise a carbonyl-containing residue linked to the amine group of
3APS via an amide bond. The carbonyl-containing residue may be
cleaved in vivo by enzymes or by any other mechanism, to liberate
the amine group of 3APS.
[0157] Preferred prodrugs are composed of a carbonyl-containing
residue linked to the amine group of 3APS via an amide bond and
such carbonyl-containing group having a nucleophile such as a
carboxylic acid or alcohol, capable of internally cleaving the
amide bond. The amino acid residue may be cleaved in vivo by
enzymes, or by any other mechanism, to liberate the amine group of
3APS.
[0158] More particularly, certain aspects of the invention relate
to a compound of Formula (IX), and to pharmaceutically acceptable
salts, esters or solvates thereof:
##STR00073##
[0159] wherein,
[0160] R.sup.8 is a substituted or unsubstituted group selected
from C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl,
C.sub.5-C.sub.15heteroaryl; and
[0161] R.sup.9 is a hydrogen or a substituted or unsubstituted
C(O)(C.sub.1-C.sub.6alkyl), C(O)NH.sub.2,
C(O)NH(C.sub.1-C.sub.6alkyl), or C(O)N(C.sub.1-C.sub.6alkyl).sub.2;
or R.sup.8 and R.sup.9 are taken together with the adjacent carbon
atom to form a substituted or unsubstituted
C.sub.3-C.sub.12heterocycloalkyl.
[0162] In one embodiment, R.sup.8 is a substituted
C.sub.1-C.sub.12alkyl. In another embodiment, R.sup.8 is a
C.sub.1-C.sub.12alkyl substituted with a substituent selected from
hydroxycarbonyl, alkoxycarbonyl, alkylcarbonyloxy, substituted or
unsubstituted 2-hydroxyphenyl, substituted or unsubstituted
2-alkylcarbonyloxyphenyl group or combinations thereof. In another
embodiment, R.sup.8 is a substituted or unsubstituted benzyl group.
In a further embodiment, R.sup.8 is selected from the groups
depicted in Table 3.
[0163] In one embodiment, the compound of the invention is a
compound of Formula IX, wherein R.sup.9 is H. In another
embodiment, the compound of the invention is a compound of Formula
IX, wherein R.sup.8 and R.sup.9 are taken together with the
adjacent carbon atom to form a substituted or unsubstituted
C.sub.3-C.sub.12heterocycloalkyl. In another embodiment, the
compound of the invention is a compound of Formula IX, wherein
R.sup.8 and R.sup.9 are taken together with the adjacent carbon
atom to form a substituted or unsubstituted phthalimide. In another
embodiment, the compound of the invention is a compound of Formula
IX, wherein R.sup.8 and R.sup.9 are taken together with the
adjacent carbon atom to form a substituted or unsubstituted
C.sub.3-C.sub.12heterocycloalkyl, wherein said heterocycle is other
than phthalimide. In another embodiment, the invention provides
compounds of Formula IX, wherein when R.sup.9 is H, then R.sup.8 is
other than benzyl, phenyl, 3-pyridinyl, 3-N-methylpyridinium,
methyl, trifluoromethyl, pentafluoroethyl, pentafluorophenyl, and
t-butyl. In another embodiment, the invention provides compounds of
Formula IX, wherein when R.sup.9 is H, then R.sup.8 is other than
n-butyl, butyl, n-propyl, i-propyl, vinyl, 2-propenyl,
2-(1-decenyl), 2-(1-dodecenyl), 1-(8-undecenyl), octyl, decyl,
undecyl, tridecyl, pentadecyl, heptadecyl,
4-(N-oxy-2,2,6,6-tetramethylpiperidinyl),
5-(1,3-dihydro-1,3-dioxo-2-benzofuranyl), 4-nitrophenyl, and
3-phenoxyphenyl. In another embodiment, the invention provides
compounds of Formula IX, wherein R.sup.8 is selected from n-butyl,
i-butyl, n-propyl, i-propyl, vinyl, 2-propenyl, 2-(1-decenyl),
2-(1-dodecenyl), 1-(8-undecenyl), octyl, decyl, undecyl, tridecyl,
pentadecyl, heptadecyl, 4-(N-oxy-2,2,6,6-tetramethylpiperidinyl),
5-(1,3-dihydro-1,3-dioxo-2-benzofuranyl), 4-nitrophenyl, and
3-phenoxyphenyl. In yet another embodiment, the invention provides
a compound of Formula IX, wherein when R.sup.9 is H, then
R.sup.8C(O) is other than a 24-oxocholan-24-yl. In yet another
embodiment, the invention provides a compound of Formula IX,
wherein when R.sup.9 is H, then R.sup.8C(O) is other than
(3.alpha.,5.beta.)-3-hydroxy-24-oxocholan-24-yl,
(3.alpha.,5.beta.,12.alpha.)-3,12-dihydroxy-24-oxocholan-24-yl,
(3.alpha.,5.beta.,7.alpha.)-3,7-dihydroxy-24-oxocholan-24-yl, or
(3.alpha.,5.beta.3,7.alpha.,12.alpha.)-3,7,12-trihydroxy-24-oxocholan-24--
yl. In yet another embodiment, the invention provides a compound of
Formula IX, wherein R.sup.8C(O) is selected from
(3.alpha.,5.beta.)-3-hydroxy-24-oxocholan-24-yl,
(3.alpha.,5.beta.,12.alpha.)-3,12-dihydroxy-24-oxocholan-24-yl,
(3.alpha.,5.beta.,7.alpha.)-3,7-dihydroxy-24-oxocholan-24-yl, and
(3.alpha.,5.beta.,7.alpha.,12.alpha.)-3,7,12-trihydroxy-24-oxocholan-24-y-
l.
[0164] The invention pertains to both salt forms and acid/base
forms of the compounds of the invention. For example, the invention
pertains not only to the particular salt forms of compounds shown
herein as salts, but also the invention includes other
pharmaceutically acceptable salts, and the acid and/or base form of
the compound. The invention also pertains to salt forms of
compounds shown herein. Compounds of the invention are also shown
in Table 3 below.
TABLE-US-00003 TABLE 3 Exemplary non-amino acid amide prodrugs
according to the invention ID Structure B1 ##STR00074## B2
##STR00075## B3 ##STR00076## B4 ##STR00077## B5 ##STR00078## B6
##STR00079## B7 ##STR00080## B8 ##STR00081## B9 ##STR00082## B10
##STR00083## B11 ##STR00084## B12 ##STR00085## B13 ##STR00086## B14
##STR00087##
II-C. Carbohydrate-Derived Prodrugs
[0165] In some preferred embodiments, the compounds of the
invention are carbohydrate-derived prodrugs that will yield or
generate 3APS once administered in a human. Preferred prodrugs
according to the invention disclosed herein comprise a carbohydrate
or a polyol analog residue linked to the amine group of 3APS via a
linkage, e.g. an amide, a carbamate, a urea, or a cleavable alkyl
group. In one embodiment, the carbohydrate-derived moiety is, for
example, a carbohydrate derivative such as hexose, pentose, a
carbohydrate-derived polyol, inositol or an inositol-derived
moiety, a carbohydrate-derived carboxylic acid, ascorbic acid,
nucleic acid, or nucleotide. The linkage and/or
carbohydrate-derived residue may be cleaved in vivo by enzymes or
by any other mechanism, to liberate the amine group of 3APS.
[0166] More particularly, certain aspects of the invention relate
to a compound of Formula (X), and to pharmaceutically acceptable
salts, esters or solvates thereof:
##STR00088##
wherein,
[0167] R.sup.10 is a residue of a carbohydrate, a carbohydrate
derivative or a carbohydrate-derived polyol, e.g., a C.sub.5-6
saturated or partially or completely unsaturated cycloalkyl group,
optionally and preferably containing an --O-- group, which is
substituted by 3 to 5 substituents, each independently selected
from --OH, --OAc, --CH.sub.2OH, --OCH.sub.3, --CH.sub.2OAc and
.dbd.O.
[0168] L is a linking moiety or is absent, e.g., an alkyl group,
which may be saturated or unsaturated, preferably a lower alkyl
group, which is optionally interrupted by one or more --O-- and/or
--NH-- groups, and is optionally substituted by one or more .dbd.O,
--OH, and/or --NH.sub.2 groups.
[0169] In one embodiment, the invention provides a compound of
Formula X, wherein when L is absent, then R.sup.10 is other than
2-deoxy-2-D-glucose.
[0170] The invention pertains to both salt forms and acid/base
forms of the compounds of the invention. For example, the invention
pertains not only to the particular salt forms of compounds shown
herein as salts, but also the invention includes other
pharmaceutically acceptable salts, and the acid and/or base form of
the compound. The invention also pertains to salt forms of
compounds shown herein. Compounds of the invention are also shown
in Table 4A below.
TABLE-US-00004 TABLE 4A Exemplary carbohydrate-derived prodrugs
according to the invention ID Structure S1 ##STR00089## S2
##STR00090## S3 ##STR00091## S4 ##STR00092## S5 ##STR00093## S6
##STR00094## S7 ##STR00095## S8 ##STR00096## S9 ##STR00097## S10
##STR00098## S11 ##STR00099## S12 ##STR00100## S13 ##STR00101## S14
##STR00102## S15 ##STR00103## S16 ##STR00104## S17 ##STR00105##
II-D. Other Prodrugs
[0171] In some preferred embodiments, the compounds of the
invention are N-hydroxy prodrugs and derivatives, cyclic
double-protected prodrugs, precursors of 3APS, as prodrugs that
will yield or generate 3APS once administered in a human.
a) N-hydroxy-Derived Prodrugs
[0172] More particularly, certain aspects of the invention relate
to a compound of Formula (XI), and to pharmaceutically acceptable
salts, esters or solvates thereof:
##STR00106##
wherein,
[0173] R.sup.11 is a hydrogen or a substituted or unsubstituted
group selected from C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl,
C.sub.5-C.sub.15heteroaryl, C(O)R.sup.12, and C(O)OR.sup.13;
and
[0174] R.sup.12 and R.sup.13 are independently selected from
substituted or unsubstituted C.sub.1-C.sub.12alkyl,
C.sub.2-C.sub.12alkenyl, C.sub.2-C.sub.12alkynyl,
C.sub.3-C.sub.15cycloalkyl, C.sub.3-C.sub.15heterocycloalkyl,
C.sub.6-C.sub.15aryl, C.sub.5-C.sub.15heteroaryl.
[0175] In one embodiment, the invention provides compounds of
Formula XI, wherein R.sup.11 is other than a hydroxyl.
[0176] Compounds of the invention include compounds:
##STR00107##
b) Cyclic Double-Protected Prodrugs
[0177] More particularly, certain aspects of the invention relate
to a compound of Formula (XII), and to pharmaceutically acceptable
salts, esters or solvates thereof:
##STR00108##
wherein,
[0178] D is a carbonyl, an amino acid residue, or a substituted
methylene group; and
[0179] X is selected from O, NH, and S.
[0180] More particularly, certain aspects of the invention relate
to a compound of Formula (XII-A), and to pharmaceutically
acceptable salts, esters or solvates thereof:
##STR00109##
wherein,
[0181] R.sup.14 is a substituted or unsubstituted group selected
from C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl,
C.sub.5-C.sub.15heteroaryl.
[0182] Compounds of the invention include the following
compounds:
##STR00110##
c) Imine Prodrugs
[0183] More particularly, certain aspects of the invention relate
to a compound of Formula (XIII), and to pharmaceutically acceptable
salts, esters or solvates thereof:
##STR00111##
wherein,
[0184] R.sup.15 and R.sup.16 are independently selected from a
hydrogen or a substituted or unsubstituted group selected from
C.sub.1-C.sub.12alkyl, C.sub.2-C.sub.12alkenyl,
C.sub.2-C.sub.12alkynyl, C.sub.3-C.sub.15cycloalkyl,
C.sub.3-C.sub.15heterocycloalkyl, C.sub.6-C.sub.15aryl, and
C.sub.5-C.sub.15heteroaryl.
[0185] In one embodiment, the invention provides compounds of
Formula XIII, wherein when both R.sup.15 and R.sup.16 are
substituted or unsubstituted aryl, then at least one of R.sup.15
and R.sup.16 is substituted with a hydroxyl group at the ortho
position. In another embodiment, the invention provides compounds
of Formula XIII, wherein when both R.sup.15 and R.sup.16 are
substituted or unsubstituted aryl, then none of R.sup.15 and
R.sup.16 is substituted with a hydroxyl group at the ortho
position.
[0186] Compounds of the invention include the following
compounds:
##STR00112##
[0187] In some preferred embodiments, the compounds of the
invention comprise a combination of any of the prodrugs described
herein in sections II-A to II-D, as prodrugs that will yield or
generate SAPS once administered in a human. The invention further
relates to sulfonic acid precursors of any of the prodrugs
mentioned in Sections II-A to II-D, including sulfonate esters,
sulfonamides, sulfinic acids, sulfides, disulfides and the
like.
II-E. Oligomers and Gemini Dimers
[0188] In a further embodiment the compound of the Formula I may
comprise two or more 3APS molecules linked together. Therefore,
another aspect of the invention relates to polymers of 3APS, i.e.,
a molecule comprising, or consisting essentially of, or consisting
of two or more molecules of 3APS linked together with cleavable
linkage. Thus, another aspect of the invention relates to a
compound of the Formula I-P:
A-(L.sup.x-A).sub.p-L.sup.x-A (I-P)
as well as pharmaceutically acceptable salts, esters, metabolites,
and solvates thereof, where:
[0189] A is 3-amino-1-propanesulfonic acid moiety;
[0190] L.sup.x is a cleavable linkage for covalently and
dissociably coupling together two adjacent 3APS moieties, and
[0191] p is 0, or an integer number which may vary from 1 to 5,
e.g. 2, 3, 4, or 5.
[0192] Those skilled in the art will readily understand that there
can be a great number of possible variations or orientations for
coupling together three or more SAPS moieties (the number of
possibilities being 2.sup.n-1, n being equal to 3 for a trimer (4
possibilities), n=4 for tetramer (8 possibilities), etc). Indeed,
as exemplified with more details hereinafter with gemini dimers,
such connections could be made via the NH.sub.2 group or the
SO.sub.3H group of the 3APS molecule. For instance, for a trimer of
3APS (i.e., 3 molecules of 3APS), there would be 4 different
possibilities:
1) .diamond-solid.--*.diamond-solid.--*.diamond-solid.--; 2)
.diamond-solid.--*.diamond-solid.--*--.diamond-solid.; 3)
.diamond-solid.--*--.diamond-solid.*.diamond-solid.--; 4)
.diamond-solid.--*--.diamond-solid.*--.diamond-solid.; the symbol
".diamond-solid." representing the NH.sub.2 group of the 3APS
molecule, the symbol "--" the SO.sub.3H group of the 3APS molecule,
and the symbol "*" representing the position of the linkage.
[0193] Alternatively, the invention relates to a compound of
Formula I-P2:
L.sup.y(A).sub.m (I-P2)
and pharmaceutically acceptable salts, esters, and solvates
thereof, where:
[0194] m is an integer from 2 to 5;
[0195] A is 3-amino-1-propanesulfonic acid moiety;
[0196] L.sup.y is a multivalent carrier moiety for covalently and
dissociably coupling from two to five A moieties, either at the
amino or sulfonic acid end of A.
[0197] In preferred embodiments, the compounds of the Formula I-P
comprise or are
[0198] "Gemini dimers" i.e., they comprise two 3APS molecules
linked together with a cleavable linkage. Thus, in another aspect;
the invention relates to compounds of Formula I-C (and salts,
esters and solvates thereof):
##STR00113##
wherein, L.sup.3 is bivalent linker which connects two molecules of
3APS at their amino groups either using the same or different
linkages as defined herein, including, but not limited to, amide
linkage and carbamate linkage.
[0199] In another aspect, the invention relates to compounds of
Formula I-D (and salts, esters, and solvates thereof):
##STR00114##
wherein, L.sup.4 is a bivalent linker which connects two molecules
of 3APS at their sulfonic acid groups either using the same or
different linkages as defined herein, including, but not limited
to, ester linkage or anhydride linkage where X is oxygen, or
sulfonamide linkage where X is nitrogen (NH, or NR), or
thiosulfonate linkage where X is sulfur. P is hydrogen or a
N-protecting group as defined herein.
[0200] In another aspect, the invention relates to compounds of
Formula I-E (and salts, esters, and solvates thereof):
##STR00115##
wherein, L.sup.5 is a bivalent linker which connects two molecules
of 3APS, at amino group in one 3APS using a linkage as defined in
Formula I-C, and at sulfonic acid group in the other 3APS using a
linkage as defined in Formula I-D. P is hydrogen or a N-protecting
group as defined herein.
[0201] In preferred embodiment, the linker L.sup.x, L.sup.3,
L.sup.4, or L.sup.5 , or the carrier moiety L.sup.y are selected
such that the two, three, four or five linked 3APS moieties may be
converted in vitro or in vivo, directly or indirectly, to release
two, three, four or five pharmaceutically active 3APS molecules.
The capability of releasing the parent 3APS molecule(s) may be
tested and, in many cases, it can be predicted. More preferably,
the linker is designed to bind the 3APS molecules via their
nitrogen atoms (for improved protection against first pass
metabolism), but as exemplified hereinbefore, it is also possible
to bind the 3APS molecules via the oxygen atom of their sulfonate
group (e.g., through an ester-type of linkage) or via their sulfur
atom (e.g., sulfonamide linked dimers). Various permutations of the
above are also possible. Those skilled in the art will be capable
to select proper linkers and linkage site and test the resulting
product for efficacy and for capability of cleavage under various
chemical and/or biological conditions. Compounds of the invention
are also shown in Table 4B below.
TABLE-US-00005 TABLE 4B Exemplary gemini dimers according to the
invention ID Structure G1 ##STR00116## G2 ##STR00117## G3
##STR00118## G4 ##STR00119## G5 ##STR00120## G6 ##STR00121## G7
##STR00122## G8 ##STR00123## G9 ##STR00124## G10 ##STR00125## G11
##STR00126##
[0202] The invention pertains to both salt forms and acid/base
forms of the compounds of the invention. For example, the invention
pertains not only to the particular salt forms of compounds shown
herein as salts, but also the invention includes other
pharmaceutically acceptable salts, and the acid and/or base form of
the compound. The invention also pertains to salt forms of
compounds shown herein.
III. Synthesis of the Compounds of the Invention
[0203] In general, all compounds of the present invention may be
prepared by the methods illustrated in the Examples hereinafter
and/or other conventional methods, using readily available and/or
conventionally preparable starting materials, reagents and
conventional synthesis procedures. In these reactions, it is also
possible to make use of variants which are in themselves known, but
are not mentioned here. Certain novel and exemplary methods of
preparing the inventive compounds are described in the
Exemplification section. Such methods are within the scope of this
invention. Functional and structural equivalents of the compounds
described herein and which have the same general properties,
wherein one or more simple variations of substituents are made
which do not adversely affect the essential nature or the utility
of the compound are also included.
[0204] More particularly, the amino acid prodrugs of the present
invention may be prepared by the methods illustrated in Example 1-A
hereinafter, and in general reaction schemes such as, for example,
described in Schemes 1 and 2, or by modifications thereof.
[0205] The carbamate prodrugs of the present invention may be
prepared by the methods illustrated in Example 1-B hereinafter, or
by modifications thereof.
[0206] The non-amino acid prodrugs of the present invention may be
prepared by the methods illustrated in Example 1-C hereinafter, and
in the general reaction schemes such as, for example, the amide
coupling steps described in Schemes 1 and 2, or by modifications
thereof.
[0207] The carbohydrate-derived prodrugs may be prepared by the
methods illustrated in Example 1-D hereinafter, or by known
coupling reactions depending on the linkage used (carbamate, urea,
amide, and the like), or by modifications thereof.
[0208] The N-hydroxy prodrugs and their derivatives may be prepared
by oxidation of the amine group, and by alkylating such N-hydroxy
group when desired. The procedures to accomplish these reactions
are readily available and known to the skilled artisan.
[0209] The cyclic double-protected prodrugs are prepared according
to standard procedures for the cyclization of such groups,
depending on the D and X groups used.
[0210] The compounds of the present invention may be readily
prepared in accordance with the synthesis schemes and protocols
described herein, as illustrated in the specific procedures
provided. However, those skilled in the art will recognize that
other synthetic pathways for forming the compounds of this
invention may be used, and that the following is provided merely by
way of example, and is not limiting to the present invention. See,
e.g., "Comprehensive Organic Transformations" by R. Larock, VCH
Publishers (1989). It will be further recognized that various
protecting and deprotecting strategies will be employed that are
standard in the art (See, e.g., "Protective Groups in Organic
Synthesis" by Greene and Wuts (1991)). Those skilled in the
relevant arts will recognize that the selection of any particular
protecting group (e.g., amine, hydroxyl, thio, and carboxyl
protecting groups) will depend on the stability of the protected
moiety with regards to the subsequent reaction conditions and will
understand the appropriate selections.
[0211] Further illustrating the knowledge of those skilled in the
art is the following sampling of the extensive chemical literature:
"Chemistry of the Amino Acids" by J. P. Greenstein and M. Winitz,
John Wiley & Sons, Inc., New York (1961); "Advanced Organic
Chemistry: Reactions, Mechanisms, and Structure" by J. March, 4th
Edition, John Wiley & sons (1992); T. D. Ocain, et al., J. Med.
Chem., 31, 2193-99 (1988); E. M. Gordon, et al., J. Med. Chem. 31,
2199-10 (1988); "Practice of Peptide Synthesis" by M. Bodansky and
A. Bodanszky, Springer-Verlag, New York (1984); "Asymmetric
Synthesis: Construction of Chiral Molecules Using Amino Acids" by
G. M. Coppola and H. F. Schuster, John Wiley & Sons, Inc., New
York (1987); "The Chemical Synthesis of Peptides" by J. Jones,
Oxford University Press, New York (1991); and "Introduction of
Peptide Chemistry" by P. D. Bailey, John Wiley & Sons, Inc.,
New York (1992).
[0212] The synthesis of compounds of the invention is preferably
carried out in a solvent. Suitable solvents are liquids at ambient
room temperature and pressure or remain in the liquid state under
the temperature and pressure conditions used in the reaction. The
choice of solvent is within the general skills of the skilled
artisan and will depend on the reaction conditions, such,
temperature, the nature of the reagents and starting material,
solubility and stability of the reagents and starting material, the
type of reaction, and the like. Depending on the circumstances,
solvents may be distilled or degassed. Solvents may be, for
example, aliphatic hydrocarbons (e.g, hexanes, heptanes, ligroin,
petroleum ether, cyclohexane, or methylcyclohexane) and halogenated
hydrocarbons (e.g., methylenechloride, chloroform,
carbontetrachloride, dichloroethane, chlorobenzene, or
dichlororbenzene); aromatic hydrocarbons (e.g., benzene, toluene,
tetrahydronaphthalene, ethylbenzene, or xylene); ethers (e.g.,
diglyme, methyl-tert-butyl ether, methyl-tert-amyl ether,
ethyl-tert-butyl ether, diethylether, diisopropylether,
tetrahydrofuran or methyltetrahydrofurans, dioxane,
dimethoxyethane, or diethyleneglycol dimethylether); amides (e.g.,
N,N-dimethylformamide, N,N-dimethylacetamide); nitriles (e.g.,
acetonitrile); ketones (e.g., acetone); esters (e.g., methyl
acetate or ethyl acetate); alcohols (e.g., methanol, ethanol,
isopropanol); water and mixtures thereof.
[0213] "Activated esters" and equivalent expressions may be
represented by the formula COX, where X is a leaving group, typical
examples of which include N-hydroxysulfosuccinimidyl and
N-hydroxysuccinimidyl groups; aryloxy groups substituted with
electron-withdrawing groups (e.g., p-nitro, pentafluoro,
pentachloro, p-cyano, or p-trifluoromethyl); and carboxylic acids
activated by a carbodiimide or other conventional coupling reagents
to form an anhydride or mixed anhydride, e.g., --OCOR.sup.a or
--OCNR.sup.aNHR.sup.b, where R.sup.a and R.sup.b are independently
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.8 alkyl (e.g., cyclohexyl),
C.sub.1-C.sub.6 perfluoroalkyl, or C.sub.1-C.sub.6 alkoxy groups.
An activated ester may be formed in situ or may be an isolable
reagent. The ester leaving group may be, for example,
sulfosuccinimidyl esters, pentafluorothiophenol esters,
sulfotetrafluorophenol, substituted or unsubstituted
C.sub.1-C.sub.6 alkyl (such as methyl, ethyl, propyl, isopropyl,
butyl, isobutyl, sec-butyl, tert-butyl, pentyl, or hexyl), or
substituted or unsubstituted C.sub.6-C.sub.14 aryl or heterocyclic
groups, such as 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl,
2,2-dibromoethyl, 2,2,2-trichloroethyl, 3-fluoropropyl,
4-chlorobutyl, methoxymethyl, 1,1-dimethyl-1-methoxymethyl,
ethoxymethyl, N-propoxymethyl, isopropoxymethyl, N-butoxymethyl,
tert-butoxymethyl, 1-ethoxyethyl, 1-methyl-1-methoxyethyl,
1-(isopropoxy)ethyl, 3-methoxypropyl-4-methoxybutyl,
fluoromethoxymethyl, 2,2,2-trichloroethoxymethyl,
bis(2-chloroethoxy)methyl, 3-fluoropropoxymethyl,
4-chlorobutoxyethyl, dibromomethoxyethyl, 2-chloroethoxypropyl,
fluoromethoxybutyl, 2-methoxyethoxymethyl, ethoxymethoxyethyl,
methoxyethoxypropyl, methoxyethoxybutyl, benzyl, phenethyl,
3-phenylpropyl, 4-phenylbutyl, .alpha.-naphthylmethyl,
.beta.-naphthylmethyl, diphenylmethyl, triphenylmethyl,
.alpha.-naphthyldipheylmethyl, 9-anthrylmethyl, 4-methylbenzyl,
2,4,6-trimethylbenzyl, 3,4,5-trimethylbenzyl, 4-methoxybenzyl,
4-methoxyphenyldiphenylmethyl, 2-nitrobenzyl, 4-nitrobenzyl,
4-chlorobenzyl, 4-bromobenzyl, 4-cyanobenzyl,
4-cyanobenzyldiphenylmethyl, or bis(2-nitrophenyl)methyl
groups.
III. Exemplary Synthesis of Amino Acid Prodrugs According to the
Invention
[0214] The following schemes are for illustration purposes and are
not intended to be limiting. The coupling of
3-amino-1-propanesulfonic acid with a first amino acid may be
generally represented by Scheme 1:
##STR00127##
wherein R.sup.1, R.sup.2 and R.sup.4 are as previously disclosed,
R.sup.Z is R.sup.3 or a protecting group, and X is the leaving
group of an activated ester.
[0215] In Scheme 1, a monoamino acid prodrug of
3-amino-1-propanesulfonic acid is produced by reacting its free
amino group (or a protected sulfonate ester variant) with an
activated ester of the desired amino acid (which may be
N-protected). Group C(O)X of the activated ester may be an acyl
halide, mixed anhydride, succinimide ester, or may be a carboxylic
acid activated by a peptide coupling agent (e.g., carbodiimides
(such as EDC
(1-(3-dimethylaminopropyl)-3-diisopropylethylcarbodiimide)) and
uroniums (such as HATU
(O-(7-azabenzotriazol-1-yl)-N,N,N',N'-diisopropylethyluronium
hexafluorophosphate))), in the presence of a base (e.g., amines
(such as DIPEA (N,N-diisopropylethylamine), hydroxides (such as
sodium hydroxide), carbonates (such as potassium carbonate), etc),
and optionally a catalyst (e.g. 4-(dimethylamino)pyridine (DMAP),
1-hydroxybenzotriazole (HOBt)). The choice of base and catalyst
will depend mainly on the nature of the activated ester.
[0216] At this stage, protecting groups (R.sup.z on amine or
protecting groups present on heteroatoms in R.sup.1 and R.sup.2
groups) may be removed. Protecting groups on heteroatoms other than
on the amine may not be removed if further amino acid couplings are
necessary (see Scheme 2). Protecting groups of oxygen atoms may
include benzyl and silyl ethers, acetals and esters, protecting
groups of nitrogen may include carbamates and fluorene derivatives.
They are cleaved by widely used procedures (see, for example,
Greene and Wuts (1991), supra).
##STR00128##
wherein R.sup.1', R.sup.2' and R.sup.4' are defined respectively as
R.sup.1, R.sup.2 and R.sup.4 but may or may not be the same as
R.sup.1, R.sup.2 and R.sup.4 in the above scheme, and R.sup.1,
R.sup.2, R.sup.z and X are as previously disclosed.
[0217] Scheme 2 is used to produce prodrugs comprising two or more
amino acids attached to 3APS. Coupling conditions are generally the
same as described for Scheme 1. Subsequent amino acids are added in
the same manner, with a deprotection of the amine group between
each coupling step. If other protecting groups are present on
heteroatoms of the residues, they may be removed during a last
chemical step.
[0218] In general, after completion of the reaction, the product is
isolated from the reaction mixture according to standard
techniques. For example, the solvent is removed by evaporation or
filtration if the product is solid, optionally under reduced
pressure. After the completion of the reaction, water may be added
to the residue to make the aqueous layer acidic or basic and the
precipitated compound filtered, although care should be exercised
when handling water-sensitive compounds. Similarly, water may be
added to the reaction mixture with a hydrophobic solvent to extract
the target compound. The organic layer may be washed with water,
dried over anhydrous magnesium sulfate or sodium sulfate, and the
solvent is evaporated to obtain the target compound. The target
compound thus obtained may be purified, if necessary, e.g., by
recrystallization, reprecipitation, chromatography, or by
converting it to a salt by addition of an acid or base.
IV. Alternate Routes and Vehicles for Delivering SAPS by Minimizing
or Lessening Hepatic First-Pass Metabolism
[0219] As indicated hereinbefore, an aspect of the invention
concerns new routes of administration (e.g. transdermally,
subcutaneously, intranasally, etc.) and new pharmaceutical vehicles
(e.g. patches, implants, spray, formulations (including for oral
administration)) for lessening hepatic first-pass metabolism of
3APS.
Transdermal Drug Delivery Devices
[0220] The delivery of drugs by the transdermal route is an area of
increasing interest and offers the advantage of allowing a
prolonged, steady input of drug into the blood. Transdermal
delivery of SAPS is one preferred embodiment of the invention
because it could avoid hepatic first-pass metabolism that is
associated with administration of 3APS, and thus increase the
therapeutic effectiveness of 3APS. Transdermal delivery may also
help avoid the pain associated with injections, and may increase
dosage compliance.
[0221] Accordingly, certain aspects of the present invention relate
to a method for the delivery of a compound according to the
invention, preferably 3APS, to improve the effectiveness of the
compound in the treatment of cognitive disorders. The invention
further relates to a method of delivering a compound according to
the invention, preferably 3APS, wherein the compound may be
administered in a transdermal patch.
[0222] Transdermal drug delivery devices according to the invention
can be manufactured using techniques and components well known to
the skilled artisan. Transdermal drug delivery devices typically
involve includes a backing layer, which may optionally be composed
of a pigmented polyester film, a drug reservoir, a microporous
membrane that controls the rate of delivery of the drug from the
system to the skin surface, and an adhesive formulation to attach
the delivery system to a subject. Optionally, the adhesive
formulation may include the drug, thus providing a more immediate
bolus of the compound upon application of the patch to a
subject.
[0223] Transdermal drug delivery devices also typically involve a
carrier (such as a liquid, gel, or solid matrix, or a pressure
sensitive adhesive) into which the drug to be delivered is
incorporated. The drug-containing carrier is then placed on the
skin and the drug, along with any adjuvants and excipients, is
delivered to the skin. Typically the portions of the carrier that
are not in contact with the subject's skin are covered by a
backing. The backing serves to protect the carrier (and the
components contained in the carrier, including the drug) from the
environment and prevents loss of the ingredients of the drug
delivery device to the environment. Because hydration of the
stratum corneum is known to enhance transport of certain drugs
across the skin, it is sometimes desirable that the backing have a
relatively low moisture vapor transmission rate in order to retain
moisture at the site covered by the drug delivery device. In order
to maintain the health of the covered skin during long term wear
(e.g., for periods in excess of a day) by allowing the skin to
breath, it is also desirable that the backing have relatively high
permeability to oxygen. Further, as the backing is in contact with
the components of the carrier, including the drug and any adjuvants
and excipients, it is important that the backing be stable to such
components in order that the backing retains its structural
integrity, tensile strength, and conformability to the skin. It is
also desirable that the backing not absorb drug or other excipients
from the carrier. In connection with the preparation of certain
reservoir-type transdermal drug delivery devices, it is also
desirable for the backing to be heat sealable at a relatively low
temperature to itself and to a variety of other polymeric
substrates. Backing materials that have found use in transdermal
drug delivery devices include metal foils, metalized plastic films,
and single layered and multilayered polymeric films (see U.S. Pat.
No. 5,264,219).
[0224] Membranes useful in the construction of a transdermal patch
are known in the art and include, but are not limited to,
CoTran.TM. membranes commercially available from 3M, such as the
COTRAN.TM. 9701, 9702, 9705, 9706, 9715, 9716, 9726, and COTRAN.TM.
9728 membranes. Backing useful in the construction of a transdermal
patch are known in the art and include, but are not limited to,
backing material commercially available from 3M, such as COTRAN.TM.
and SCOTCHPAK.TM. backings. Likewise, liners are well known in the
art and may be obtained from a number of commercial sources.
Optionally, a gelling agent may be optionally added at up to 20% by
volume. Gelling agents, include, but are not limited to:
crosslinked acrylic acid polymers, such as the "carbomer" family of
polymers, e.g., carboxypolyalkylenes that may be obtained
commercially under the tradename CARBOPOL.TM.; hydrophilic
polymers, such as polyethylene oxides,
polyoxyethylene-polyoxypropylene copolymers and polyvinylalcohol;
cellulosic polymers, such as hydroxypropyl cellulose, hydroxycthyl
cellulose, hydroxypropyl methylcellulose, hydroxypropyl
methylcellulose phthalate, and methyl cellulose; gums such as
tragacanth and xanthan gum; sodium alginate; and gelatin.
[0225] Those skilled in the art will readily identified the proper
combination and/or concentration of backing layer, drug reservoir,
membrane, carrier, backing, penetration enhancer, gelling agent,
etc. If necessary, one could refer to the numerous publications on
the subject, including the patent literature such as EP 1 602 367,
US 2005/019384, US 2005/0074487 and US 2005/175680 all describing
transdermal drug delivery devices and associated. For instance, the
absorption through human skin of a drug may be used to determine
the feasibility of transdermal delivery with a particular carrier
or vehicle. For example, penetration of a compound according to the
invention, preferably 3APS, across human epidermis may be measured
in vitro using glass diffusion cells. Therefore, optimization of
the compound's absorption is achieved by use of formulations
disclosed herein and known in the art as penetration enhancers.
Additional well known tests and assays include permeability rate
studies, absorption studies, diffusion assays, time-course
profiling for penetration across human epidermis, irritancy
studies, etc.
[0226] Clinical studies indicate that an oral dose of a SAPS of
about 100 and/or 150 mg bid may produce a beneficial treatment for
cognitive disorders, such as AD. Since transdermal administration
of a compound according to the invention, preferably 3APS, is
believed to be subject to reduced first-pass metabolism, the dosage
of a 3APS may be reduced when administered transdermally. On the
other hand, transdermal administration could be helpful to increase
the dosage of 3APS by avoiding common side effects such as
gastrointestinal irritation associated with an oral administration
of that drug.
[0227] A benefit of a transdermal dosage form includes improved
subject compliance, due to the possibility of reduced
administrations. For example, a transdermal patch of the invention
may be formulated so as to provide one, two, three, four, five, six
or seven days of medication. In an exemplary embodiment, the
transdermal patch provides medication for about three days, before
it is desirable to replace the patch. In another exemplary
embodiment, the transdermal patch provides seven days of
medication, before it is desirable to replace the patch. In
addition, the transdermal dosage may be formulated with any
desirable dosage of a compound according to the invention. For
example, the transdermal dosage may provide the equivalent dosage
to oral administration of 100 mg bid, 150 mg bid, 200 mg bid, 250
mg bid, 300 mg bid, 350 mg bid, or 400 mg bid. As such, a
transdermal patch(es) having the equivalent dosage to oral
administration of 150 mg bid may be administered to a subject for a
desired period of time, for example, four weeks, and then a patch
or patches having the equivalent dosage to oral administration of
200 mg bid may be administered for a desired period of time.
[0228] Also included within the invention is a kit, which may
contain a desired supply of patches, for example, a one month
supply of transdermal patches. Optionally, a kit may be organized
into a plurality of, e.g., three, differently identified (numbered,
colored or the like) parts, wherein the contents of a first part
are initially administered, followed by administration of the
contents of a second part, which are then followed by
administration of the contents of the third part. Alternatively, a
kit may contain a combination of patches and oral formulations.
V. Subjects and Patient Populations
[0229] The term "subject" includes living organisms in which
A.beta.-amyloidosis can occur, or which are susceptible to
A.beta.-amyloid diseases, e.g., Alzheimer's disease, etc. Examples
of subjects include humans, chickens, ducks, Peking ducks, geese,
monkeys, deer, cows, rabbits, sheep, goats, dogs, cats, mice, rats,
and transgenic species thereof. The term "subject" preferably
includes animals susceptible to states characterized by neuronal
cell death, e.g. mammals, e.g. humans. The animal can be an animal
model for a disorder, e.g., a transgenic mouse with an
Alzheimer's-type neuropathology. In preferred embodiments, the
subject is a mammal, more preferably a human subject.
[0230] The term "human subject" includes humans susceptible to
benefit from SAPS administration, and more particularly those
susceptible to or diagnosed of having an amyloid-.beta. related
disease and/or suffering from a neurodegenerative disease, such as
Alzheimer's disease, Parkinson's disease, etc.
[0231] In certain embodiments of the invention, the human subject
is in need of treatment by the methods of the invention, and is
selected for treatment based on this need. A subject in need of
treatment is art-recognized, and includes subjects that have been
identified as having a disease or disorder related to
.beta.-amyloid deposition, has a symptom of such a disease or
disorder, or is at risk of such a disease or disorder, and would be
expected, based on diagnosis, e.g., medical diagnosis, to benefit
from treatment (e.g., curing, healing, preventing, alleviating,
relieving, altering, remedying, ameliorating, improving, or
affecting the disease or disorder, the symptom of the disease or
disorder, or the risk of the disease or disorder).
[0232] For example, the human subject may be a human over 30 years
old, human over 40 years old, a human over 50 years old, a human
over 60 years old, a human over 70 years old, a human over 80 years
old, a human over 85 years old, a human over 90 years old, or a
human over 95 years old. The subject may be a female human,
including a postmenopausal female human, who may be on hormone
(estrogen) replacement therapy. The subject may also be a male
human. In another embodiment, the subject is under 40 years
old.
[0233] In preferred embodiments, the subject is a human subject
having an Alzheimer's-type neuropathology. Individuals presently
suffering from Alzheimer's disease can be recognized from
characteristic dementia, as well as the presence of risk factors
described below. In addition, a number of diagnostic tests based on
cognitive and neurological testing are available for identifying
individuals who have AD. For example, individuals suffering from
Alzheimer's disease can be diagnosed by the Clinical Dementia
Rating (CDR) scale, Mini-mental State Examination (MMSE),
Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog),
or any other test known in the art, as discussed herein. Baseline
scores on suitable metrics including the MMSE and the ADAS together
with other metrics designed to evaluate a more normal population
can be used to find an at risk population. Another method for
identifying an at risk group utilizes an assay for neural thread
protein in the urine; see, e.g., Munzar et al., Neurology and
Clinical Neurophysiology, Vol. 2002, No. 1. Patients with high risk
for Alzheimer's Disease can also be selected from a population by
screening for early signs of memory loss or other difficulties
associated with pre-Alzheimer's symptomatology, a family history of
Alzheimer's Disease, patients with Mild Cognitive Impairment (MCI),
genetic risk factors, age, sex, and other features found to predict
high-risk for Alzheimer's Disease.
[0234] The term "prevention" or "preventing" is also used to
describe the administration of a compound or composition of the
invention to a subject who is at risk of (or susceptible to) such a
disease or condition. Patients amenable to treatment for prevention
of the disease or condition include individuals at risk of the
disease or condition but not showing symptoms, as well as patients
presently showing symptoms. In the case of Alzheimer's disease,
virtually anyone is at risk of suffering from Alzheimer's disease
if he or she lives long enough. Therefore, the present methods can
be administered prophylactically to the general population without
any assessment of the risk of the subject patient. But the present
methods are especially useful for individuals who do have a known
risk of Alzheimer's disease. Such individuals include those having
relatives who have experienced this disease, and those whose risk
is determined by analysis of genetic or biochemical markers,
including brain plaques diagnosed by imaging methods, e.g., MRI,
PET, SPECT etc.. Examples of such imaging methods are discussed in
Burggren et al., Current Topics in Medicinal Chemistry, vol. 2002,
no. 2, pp. 385-393, and Sair et al., Neuroradiology, vol. 46, pp.
93-104 (2002). Alzheimer's disease predisposing factors identified
or proposed in the scientific literature include, among others, a
genotype predisposing a subject to Alzheimer's disease;
environmental factors predisposing a subject to Alzheimer's
disease; past history of infection by viral and bacterial agents
predisposing a subject to Alzheimer's disease; and vascular factors
predisposing a subject to Alzheimer's disease. Genetic markers of
risk toward Alzheimer's disease include mutations in the APP gene,
particularly mutations at position 717 and positions 670 and 671
referred to as the Hardy and Swedish mutations respectively (see
Hardy et al., TINS 20, 154-158 (1997)). Other markers of risk are
mutations in the presenilin genes, PS1 and PS2, and ApoE4, family
history of AD, hypercholesterolemia or atherosclerosis. The subject
may be shown to be at risk by a diagnostic brain imaging technique,
for example, one that measures brain activity, plaque deposition,
or brain atrophy. The human subject may also be shown to be at risk
by a cognitive test such as Clinical Dementia Rating ("CDR"),
Alzheimer's disease Assessment Scale-Cognition ("ADAS-Cog"),
Disability Assessment for Dementia ("DAD") or Mini-Mental State
Examination ("MMSE") and/or by any other cognition test known in
the art.
[0235] In another embodiment, the human subject exhibits no
symptoms of Alzheimer's disease. In another embodiment, the subject
is at least 40 years of age and exhibits no symptoms of Alzheimer's
disease. In another embodiment, the human subject is at least 40
years of age and exhibits one or more symptoms of Alzheimer's
disease.
[0236] By using the methods and compounds of the present invention,
the levels of amyloid .beta. peptides in a subject's plasma or
cerebrospinal fluid (CSF) could be significantly reduced from
levels prior to treatment from about 10 to about 100 percent, or
even about 50 to about 100 percent, e.g., 15, 25, 40, 60, 70, 75,
80, 90, 95 or 99%. Accordingly, in certain embodiments, the human
subject can have an elevated level of amyloid A.beta..sub.40 and
A.beta..sub.42 peptide in the blood and/or CSF prior to a treatment
according to the present methods, e.g. A.beta..sub.40 levels of
greater than about 10 pg/mL, or greater than about 20 pg/mL, or
greater than about 35 pg/mL, or even greater than about 40 pg/mL;
and A.beta..sub.42 levels 30 pg/mL to about 200 pg/mL, or even to
about 500 pg/mL. Similarly, according to some embodiments, the
methods and compounds of the present invention help reduce the size
and/or number of A.beta. plaques or A.beta. deposits in the brain,
from about 10 to about 100 percent, or even about 50 to about 100
percent, e.g., 15, 25, 40, 60, 70, 75, 80, 90, 95 or 99%, when
compared to levels prior to treatment.
VI. Pharmaceutical Compositions
[0237] Preferably, the compounds of the invention are formulated
prior to administration into pharmaceutical compositions using
techniques and procedures well known in the art. Accordingly, in
another embodiment, the present invention relates to pharmaceutical
compositions (e.g. solutions, suspensions or emulsions) comprising
effective amounts of one or more compounds according to any of the
Formulae herein and a pharmaceutically acceptable vehicle, as well
as methods of using and manufacturing such pharmaceutical
compositions.
[0238] The pharmaceutical compositions are formulated into suitable
administration (orally, parenterally, (IV, IM, depo-IM, SC, and
depo SC), sublingually, intranasally (inhalation), intrathecally,
topically, or rectally). Suitable pharmaceutically acceptable
vehicles include, without limitation, any non-immunogenic
pharmaceutical carrier or diluent suitable for oral, parenteral,
nasal, mucosal, transdermal, topical, intrathecal, rectal,
intravascular (IV), intraarterial (IA), intramuscular (IM), and
subcutaneous (SC) administration routes, such as phosphate buffer
saline (PBS). Also, the present invention includes such compounds
which have been lyophilized and which may be reconstituted to form
pharmaceutically acceptable formulations for administration, as by
intravenous, intramuscular, or subcutaneous injection.
Administration may also be intradermal or transdermal.
[0239] The vehicle can be a solvent or dispersion medium
containing, for example, water, ethanol, polyol (for example,
glycerol, propylene glycol, and liquid polyethylene glycol, and the
like), suitable mixtures thereof, and vegetable oils. The proper
fluidity can be maintained, for example, by the use of a coating
such as lecithin, by the maintenance of the required particle size
in the case of dispersion and by the use of surfactants. Prevention
of the action of microorganisms can be achieved by various
antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In
many cases, isotonic agents are included, for example, sugars,
sodium chloride, or polyalcohols such as mannitol and sorbitol, in
the composition. Prolonged absorption of the injectable
compositions can be brought about by including in the composition
an agent which delays absorption, for example, aluminum
monostearate or gelatin.
[0240] Preferably, the compound(s) of the invention can be orally
administered. Formulations of the present invention include those
suitable for oral administration. The formulations may conveniently
be presented in unit dosage form and may be prepared by any methods
well known in the art of pharmacy. Methods of preparing these
formulations or compositions include the step of bringing into
association a compound of the present invention with a
pharmaceutically acceptable vehicle (e.g. an inert diluent or an
assimilable edible carrier)and, optionally, one or more accessory
ingredients. In general, the formulations are prepared by uniformly
and intimately bringing into association a compound of the present
invention with liquid carriers, or finely divided solid carriers,
or both, and then, if necessary, shaping the product. The amount of
the therapeutic agent in such therapeutically useful compositions
is such that a suitable dosage will be obtained.
[0241] Formulations of the invention suitable for oral
administration may be in the form of capsules (e.g. hard or soft
shell gelatin capsule), cachets, pills, tablets, lozenges, powders,
granules, pellets, dragees, e.g., coated (e.g., enteric coated) or
uncoated, or as a solution or a suspension in an aqueous or
non-aqueous liquid, or as an oil-in-water or water-in-oil liquid
emulsion, or as an elixir or syrup, or as pastilles (using an inert
base, such as gelatin and glycerin, or sucrose and acacia) or as
mouth washes and the like, each containing a predetermined amount
of a compound of the present invention as an active ingredient. A
compound of the present invention may also be administered as a
bolus, electuary or paste, or incorporated directly into the
subject's diet. Moreover, in certain embodiments these pellets can
be formulated to (a) provide for instant or rapid drug release
(i.e., have no coating on them); (b) be coated, e.g., to provide
for sustained drug release over time; or (c) be coated with an
enteric coating for better gastrointestinal tolerability.
[0242] In solid dosage forms of the invention for oral
administration the active ingredient is mixed with one or more
pharmaceutically acceptable carriers, such as sodium citrate or
dicalcium phosphate, or any of the following: fillers or extenders,
such as starches, lactose, sucrose, glucose, mannitol, or silicic
acid; binders, such as, for example, carboxymethylcellulose,
alginates, gelatin, polyvinyl pyrrolidone, sucrose or acacia;
humectants, such as glycerol; disintegrating agents, such as
agar-agar, calcium carbonate, potato or tapioca starch, alginic
acid, certain silicates, and sodium carbonate; solution retarding
agents, such as paraffin; absorption accelerators, such as
quaternary ammonium compounds; wetting agents, such as, for
example, cetyl alcohol and glycerol monostearate; absorbents, such
as kaolin and bentonite clay; lubricants, such as talc, calcium
stearate, magnesium stearate, solid polyethylene glycols, sodium
lauryl sulfate, and mixtures thereof; and coloring agents. In the
case of capsules, tablets and pills, the pharmaceutical
compositions may also comprise buffering agents. Solid compositions
of a similar type may also be employed as fillers in soft and
hard-filled gelatin capsules using such excipients as lactose or
milk sugars, as well as high molecular weight polyethylene glycols
and the like.
[0243] Peroral compositions typically include liquid solutions,
emulsions, suspensions, and the like. The pharmaceutically
acceptable vehicles suitable for preparation of such compositions
are well known in the art. Typical components of carriers for
syrups, elixirs, emulsions and suspensions include ethanol,
glycerol, propylene glycol, polyethylene glycol, liquid sucrose,
sorbitol and water. For a suspension, typical suspending agents
include methyl cellulose, sodium carboxymethyl cellulose,
tragacanth, and sodium alginate; typical wetting agents include
lecithin and polysorbate 80; and typical preservatives include
methyl paraben and sodium benzoate. Peroral liquid compositions may
also contain one or more components such as sweeteners, flavoring
agents and colorants disclosed above.
[0244] Pharmaceutical compositions suitable for injectable use
include sterile aqueous solutions (where water soluble) or
dispersions, and sterile powders for the extemporaneous preparation
of sterile injectable solutions or dispersion. In all cases, the
composition must be sterile and must be fluid to the extent that
easy syringability exists. It must be stable under the conditions
of manufacture and storage and must be preserved against the
contaminating action of microorganisms such as bacteria and fungi.
Sterile injectable solutions can be prepared by incorporating the
therapeutic agent in the required amount in an appropriate solvent
with one or a combination of ingredients enumerated above, as
required, followed by filtered sterilization. Generally,
dispersions are prepared by incorporating the therapeutic agent
into a sterile vehicle which contains a basic dispersion medium and
the required other ingredients from those enumerated above. In the
case of sterile powders for the preparation of sterile injectable
solutions, the methods of preparation are vacuum drying and
freeze-drying which yields a powder of the active ingredient (i.e.,
the therapeutic agent) plus any additional desired ingredient from
a previously sterile-filtered solution thereof.
[0245] Pharmaceutical formulations are also provided which are
suitable for administration as an aerosol, by inhalation. These
formulations comprise a solution or suspension of the desired
compound of any Formula herein or a plurality of solid particles of
such compound(s). The desired formulation may be placed in a small
chamber and nebulized. Nebulization may be accomplished by
compressed air or by ultrasonic energy to form a plurality of
liquid droplets or solid particles comprising the agents or salts.
The liquid droplets or solid particles should have a particle size
in the range of about 0.5 to about 5 microns. The solid particles
can be obtained by processing the solid agent of any Formula
described herein, or a salt thereof, in any appropriate manner
known in the art, such as by micronization. The size of the solid
particles or droplets will be, for example, from about 1 to about 2
microns. In this respect, commercial nebulizers are available to
achieve this purpose.
[0246] A pharmaceutical formulation suitable for administration as
an aerosol may be in the form of a liquid, the formulation will
comprise a water-soluble agent of any Formula described herein, or
a salt thereof, in a carrier which comprises water. A surfactant
may be present which lowers the surface tension of the formulation
sufficiently to result in the formation of droplets within the
desired size range when subjected to nebulization.
[0247] The compositions of this invention can also be administered
topically to a subject, e.g., by the direct laying on or spreading
of the composition on the epidermal or epithelial tissue of the
subject, or transdermally via a "patch". Such compositions include,
for example, lotions, creams, solutions, gels and solids. These
topical compositions may comprise an effective amount, usually at
least about 0.1%, or even from about 1% to about 5%, of an agent of
the invention. Suitable carriers for topical administration
typically remain in place on the skin as a continuous film, and
resist being removed by perspiration or immersion in water.
Generally, the carrier is organic in nature and capable of having
dispersed or dissolved therein the therapeutic agent. The carrier
may include pharmaceutically acceptable emollients, emulsifiers,
thickening agents, solvents and the like.
[0248] Other compositions useful for attaining systemic delivery of
the subject agents include sublingual, buccal and nasal dosage
forms. Such compositions typically comprise one or more of soluble
filler substances such as sucrose, sorbitol and mannitol; and
binders such as acacia, microcrystalline cellulose, carboxymethyl
cellulose and hydroxypropyl methyl cellulose. Glidants, lubricants,
sweeteners, colorants, antioxidants and flavoring agents disclosed
above may also be included. The compound(s) of the invention may
also be administered parenterally, intraperitoneally,
intraspinally, or intracerebrally. For such compositions, the
compound(s) of the invention can be prepared in glycerol, liquid
polyethylene glycols, and mixtures thereof and in oils. Under
ordinary conditions of storage and use, these preparations may
contain a preservative to prevent the growth of microorganisms.
[0249] To administer the compound(s) of the invention by other than
parenteral administration, it may be useful to coat the compound(s)
with, or co-administer the compound(s) with a material to prevent
its inactivation. For example, the compound(s) of the invention may
be administered to a subject in an appropriate carrier, for
example, liposomes, or a diluent. Pharmaceutically acceptable
diluents include saline and aqueous buffer solutions. Liposomes
include water-in-oil-in-water CGF emulsions as well as conventional
liposomes.
[0250] Pharmaceutical compositions according to the invention may
also be coated by conventional methods, typically with pH or
time-dependent coatings, such that the compound(s) of the invention
is released in the vicinity of the desired location, or at various
times to extend the desired action. Such dosage forms typically
include, but are not limited to, one or more of cellulose acetate
phthalate, polyvinylacetate phthalate, hydroxypropyl methyl
cellulose phthalate, ethyl cellulose, waxes, and shellac.
[0251] The compound(s) of the invention may be packaged as part of
a kit, optionally including a container (e.g. packaging, a box, a
vial, etc). The kit may be commercially used according to the
methods described herein and may include instructions for use in a
method of the invention. Additional kit components may include
acids, bases, buffering agents, inorganic salts, solvents,
antioxidants, preservatives, or metal chelators. The additional kit
components are present as pure compositions, or as aqueous or
organic solutions that incorporate one or more additional kit
components. Any or all of the kit components optionally further
comprise buffers.
VII. Dosage
[0252] Dosage forms, upon releasing a compound according to the
invention, can provide the corresponding 3APS upon in vivo
administration to a human patient. It is understood that
appropriate doses depend upon a number of factors within the
knowledge of the ordinarily skilled physician, veterinarian, or
researcher (e.g. see Wells et al. eds., Pharmacotherapy Handbook,
2.sup.nd Edition, Appleton and Lange, Stamford, Conn. (2000); PDR
Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition,
Tarascon Publishing, Loma Linda, Calif. (2000)). The dose(s) of the
compound(s) of the invention will vary, for example, depending upon
a variety of factors including the activity of the specific agent
employed, the age, body weight, general health, gender, and diet of
the subject, the time of administration, the route of
administration, the rate of excretion, and any drug combination, if
applicable, the effect which the practitioner desires the compound
to have upon the subject and the properties of the compounds (e.g.
bioavailability, stability, potency, toxicity, etc). Such
appropriate doses may be determined using the assays described
herein. When one or more of the compounds of the invention is to be
administered to humans, a physician may for example, prescribe a
relatively low dose at first, subsequently increasing the dose
until an appropriate response is obtained.
[0253] Exemplary doses include milligram or microgram amounts of
the compound per kilogram of subject or sample weight (e.g., about
50 micrograms per kilogram to about 500 milligrams per kilogram,
about 1 milligram per kilogram to about 100 milligrams per
kilogram, about 1 milligram per kilogram to about 50 milligram per
kilogram, about 1 milligram per kilogram to about 10 milligrams per
kilogram, or about 3 milligrams per kilogram to about 5 milligrams
per kilogram). Additional exemplary doses include doses of about 5
to about 500 mg, or about 25 to about 300 mg, or about 25 to about
200 mg, preferably about 50 to about 150 mg, more preferably about
50, about 100, about 150 mg, about 200 mg or about 250 mg, and,
preferably, daily or twice daily, or lower or higher amounts. For
comparison, exemplary doses for 3APS per se include about 2-3
milligram of 3APS per kilogram of subject (twice daily). See also
U.S. Ser. No. 11/103,656, filed on Apr. 12, 2005, which is
incorporated herein by reference.
[0254] It is generally advantageous to formulate parenteral
compositions in dosage unit form for ease of administration and
uniformity of dosage. The term "unit dosage form" refers to a
physically discrete unit suitable as unitary dosages for human
subjects and other mammals, each unit containing a predetermined
quantity of active material calculated to produce the desired
therapeutic effect, in association with a suitable pharmaceutical
vehicle. In an embodiment, the compositions according to the
invention are formulated in a unit dosage form, each dosage
containing from about 50 mg to about 500 mg, more preferably about
100 mg to about 300 mg of the compound according to the invention.
See also U.S. Ser. No. 11/103,656, filed on Apr. 12, 2005, which is
incorporated herein by reference. The specification for the dosage
unit forms of the invention may vary and are dictated by and
directly dependent on (a) the unique characteristics of the
therapeutic agent and the particular therapeutic effect to be
achieved, and (b) the limitations inherent in the art of
compounding such a therapeutic agent for the treatment of amyloid
deposition in subjects.
[0255] Administration of the compounds and compositions of the
present invention to a subject to be treated can be carried out
using known procedures, at dosages and for periods of time
effective to achieved a desired purposes (e.g. prevention or
treatment of AD, obtaining specific levels of SAPS, etc). Dosage
regimens can be adjusted to provide the optimum therapeutic
response. For example, several divided doses may be administered
daily or the dose may be proportionally reduced as indicated by the
exigencies of the therapeutic situation.
[0256] In one embodiment, the compound(s) of the invention is
administered at a therapeutically effective dosage sufficient to
inhibit amyloid deposition in a subject, preferably a human
subject. When referring to amyloid deposition a "therapeutically
effective" dosage inhibits amyloid deposition by, for example, at
least about 20%, or by at least about 40%, or even by at least
about 60%, or by at least about 80% relative to untreated
subjects.
[0257] In one embodiment, the compound(s) of the invention is
administered at a therapeutically effective dosage for the
prevention or treatment of Alzheimer's. When referring to
Alzheimer's, a "therapeutically effective" dosage stabilizes
cognitive function or prevents a further decrease in cognitive
function (i.e., preventing, slowing, or stopping disease
progression).
VII. Uses of Compounds, Composition, and Dosage Forms
[0258] Another aspect of the invention pertains to a method for
inhibiting neuronal cell death by administering an effective amount
of a compound of the present invention. In yet another aspect, the
invention pertains to a method for providing neuroprotection to a
subject having an A.beta.-amyloid related disease, e.g. Alzheimer's
disease, which includes administering an effective amount of a
compound of the present invention to the subject, such that
neuroprotection is provided. As used herein, the term
"neuroprotection" includes protection of neuronal cells of a
subject from cell death that may result in initiation of processes
such as, but not limited to: the destabilization of the
cytoskeleton; DNA fragmentation; the activation of hydrolytic
enzymes, such as phospholipase A2; activation of caspases,
calcium-activated proteases and/or calcium-activated endonucleases;
inflammation mediated by macrophages; calcium influx into a cell;
membrane potential changes in a cell; the disruption of cell
junctions leading to decreased or absent cell-cell communication;
and the activation of expression of genes involved in cell
death.
[0259] According to a preferred embodiment, the compounds and
compositions of the present invention are used for one or more of
the following: to prevent Alzheimer's disease, to treat Alzheimer's
disease, or ameliorate symptoms of Alzheimer's disease, to regulate
production of or levels of amyloid .beta. (A.beta.) peptides,
prevent, reduce, or inhibit amyloid deposition in a subject, and to
treat or prevent of amyloid-related diseases.
[0260] The compounds and pharmaceutical compositions of the
invention may act to ameliorate the course of a .beta.-amyloid
related disease using any of the following mechanisms (this list is
meant to be illustrative and not limiting): slowing the rate of
.beta.-amyloid fibril formation or deposition; lessening the degree
of .beta.-amyloid deposition; inhibiting, reducing, or preventing
amyloid fibril formation; inhibiting neurodegeneration or cellular
toxicity induced by .beta.-amyloid; inhibiting amyloid induced
inflammation; enhancing the clearance of .beta.-amyloid from the
brain; enhancing degradation of A.beta. in the brain; or favoring
clearance of amyloid protein prior to its organization in fibrils,
and decreasing the ratio of A.beta.42:A.beta.40 in the CSF or
plasma. In another embodiment, the invention pertains to a method
for improving cognition in a subject suffering from AD. The method
includes administering an effective amount of a therapeutic
compound of the invention, such that the subject's cognition is
improved. The subject's cognition can be tested using methods known
in the art such as the Clinical Dementia Rating ("CDR"),
Mini-Mental State Examination ("MMSE"), Disability Assessment for
Dementia ("DAD"), and the Alzheimer's Disease Assessment
Scale-Cognition ("ADAS-Cog"). Improvement in cognition is present
within the context of the present invention if there is a
measurable difference between the performances of subjects treated
using the methods of the invention as compared to members of a
placebo group, historical control, or between subsequent tests
given to the same subject. The invention also pertains to a method
for treating, slowing or stopping a 3-amyloid related disease
associated with cognitive impairment, by administering to a subject
an effective amount of a therapeutic compound of the invention,
wherein the annual deterioration of the subject's cognition as
measured by any of the foregoing mentioned test is improved.
[0261] It is to be understood that wherever values and ranges are
provided herein, e.g., in ages of subject populations, dosages, and
blood levels, all values and ranges encompassed by these values and
ranges, are meant to be encompassed within the scope of the present
invention. Moreover, all values in these values and ranges may also
be the upper or lower limits of a range.
VIII. Combination Therapy
[0262] In certain embodiments, the compounds and composition
according to the invention can be used in combination therapy with
at least one other therapeutic agent. The prodrug compounds
according to the invention and the at least one other therapeutic
agent(s) can act additively or, in certain embodiments,
synergistically. In certain embodiments, the compounds of the
invention can be administered concurrently with the administration
of another therapeutic agent. In certain embodiments, the compounds
of the invention can be administered prior or subsequent to
administration of another therapeutic agent. The at least one other
therapeutic agent can be effective for treating the same or
different disease, disorder, or condition.
[0263] Methods of the present invention include administration of
one or more compounds or pharmaceutical compositions of the present
invention and one or more other therapeutic agents provided that
the combined administration does not inhibit the therapeutic
efficacy of the one or more compounds of the present invention
and/or does not produce adverse combination effects.
[0264] In certain embodiments, compositions of the present
invention can be administered concurrently with the administration
of another therapeutic agent, which can be part of the same
pharmaceutical composition as, or in a different composition from,
that containing the compounds of the present invention. In certain
embodiments, compounds of the present invention can be administered
prior or subsequent to administration of another therapeutic agent.
In certain embodiments of combination therapy, the combination
therapy comprises alternating between administering a composition
of the present invention and a composition comprising another
therapeutic agent, e.g., to minimize adverse side effects
associated with a particular drug. When a compound of the present
invention is administered concurrently with another therapeutic
agent that potentially can produce adverse side effects including,
but not limited to, toxicity, the therapeutic agent can
advantageously be administered at a dose that falls below the
threshold at which the adverse side effect is elicited.
[0265] In certain embodiments, a pharmaceutical composition can
further comprise substances to enhance, modulate and/or control
release, bioavailability, therapeutic efficacy, therapeutic
potency, stability, and the like. For example, to enhance
therapeutic efficacy a compound of the present invention, the
compound can be co-administered with one or more active agents to
increase the absorption or diffusion of the compound from the
gastrointestinal tract, or to inhibit degradation of the drug in
the systemic circulation. In certain embodiments, at least one
compound of the present invention can be co-administered with
active agents having a pharmacological effect that enhance the
therapeutic efficacy of SAPS.
[0266] In certain embodiments, compounds or pharmaceutical
compositions of the present invention include, or can be
administered to a patient together with, another therapeutic drug
that may be available over-the-counter or by prescription. US
patent application No. 2005/0031651 (incorporated herein by
reference) provide a long but non-exhaustive list of "therapeutic
drugs" that can be useful, in combination, according to the
invention. Preferred therapeutic drugs to be used with the
compounds or pharmaceutical compositions of the present invention
are therapeutic drugs useful in the prevention or treatment of
Alzheimer's Disease or its symptoms, including but not limited to
donepezil (Aricept.TM.), memantine (Namenda.TM.), rivastigmine
(Exelon.TM.), Galanthamine (Reminyl.TM. and R-flurbiprofen
(Flurizan.TM.). The compounds and compositions according to the
invention could also be combined with vaccines and antibodies for
the prevention or treatment of AD.
[0267] In a further embodiment, the compounds of the invention can
be co-administered with 3APS.
IX. Standard Methods for Testing the Compounds of the
Invention.
[0268] The compounds according to the invention can be further
analyzed, tested or validated using a variety of in vitro assays,
or in vivo assays to confirm their safety, bioavailabity,
neuroprotection, their capability to deliver 3APS etc. The
following are illustrative of the type of biological assays that
can be conducted to assess the instant compounds.
i) Determination of Enzymatic Cleavage of Prodrugs In Vitro
[0269] For orally administered prodrugs, it is generally desirable
that the prodrug remains intact (i.e., uncleaved) while in the
gastrointestinal tract and be cleaved (i.e., to release the parent
drug) while in the systemic circulation. A useful level of
stability can at least in part be determined by the mechanism and
kinetics of absorption of the prodrug by the gastrointestinal
tract. A useful level of lability can at least in part be
determined by the pharmacokinetics of the prodrug and parent drug
in the systemic circulation. In general, prodrugs that are more
stable in a Caco-2 S9 and/or pancreatin assay and are more labile
in a rat plasma, human plasma, rat liver S9, and/or human liver S9
preparation can be useful as an orally administered prodrug. The
results of tests, for determining the enzymatic cleavage of
prodrugs in vitro can be used to select prodrugs for in vivo
testing.
ii) Bioavailability of Prodrugs In Vivo
[0270] Prodrugs that provide a bioavailability of the corresponding
parent drug that is greater than the bioavailability provided by an
equimolar dose of the parent drug administered to a patient by the
same route (e.g., oral administration) can be useful as therapeutic
agents. Bioavailability of the compounds of the invention and of
released SAPS can be measured in vivo (humans and laboratory
animals) using methods well known in the art. Example 3 herein
provides an exemplary method for assessing bioavailability in
mice.
iii) In Vivo Assays; Animal Models
[0271] Various animal models can be used to the efficacy and/or
potency of the compound according to the invention. For example,
certain transgenic animal models have been described, for example,
in U.S. Pat. Nos. 5,877,399; 5,612,486; 5,387,742; 5,720,936;
5,850,003; 5,877,015, and 5,811,633, and in Ganes et al., (Nature
1995, 373:523). Preferred are animal models that exhibit
characteristics associated with the pathophysiology of AD.
Administration of the compound inhibitors of the invention to the
transgenic mice described herein provides an alternative method for
demonstrating the inhibitory activity of the compounds.
Administration of the compounds in a pharmaceutically effective
carrier and via an administrative route that reaches the target
tissue in an appropriate therapeutic amount is also preferred.
iv) Toxicity
[0272] A variety of different parameters can be monitored to assess
toxicity. Examples of such parameters include, but are not limited
to, cell proliferation, monitoring activation of cellular pathways
for toxicological responses by gene or protein expression analysis,
DNA fragmentation, changes in the composition of cellular
membranes, membrane permeability, activation of components of
death-receptors or downstream signaling pathways (e.g., caspases),
generic stress responses, NF-kappaB activation and responses to
mitogens. Related assays are used to assay for apoptosis (a
programmed process of cell death) and necrosis, including cGMP
formation and NO formation.
[0273] Toxicity and therapeutic efficacy of the compound(s) and
composition(s) of the invention can be determined by standard
pharmaceutical procedures in cell cultures or experimental animals,
e.g., for determining the LD50 (the dose lethal to 50% of the
population) and the ED50 (the dose therapeutically effective in 50%
of the population). The dose ratio between toxic and therapeutic
effects is the therapeutic index and can be expressed as the ratio
LD50/ED50, and usually a larger therapeutic index is more
efficacious. While agents that exhibit toxic side effects may be
used, care should be taken to design a delivery system that targets
such agents to the site of affected tissue in order to minimize
potential damage to unaffected cells and, thereby, reduce side
effects.
v) Neuroprotection
[0274] The following are illustrative of the type of biological
assays that can be conducted to assess whether a inhibitory agent
has a protective effect against neuronal injury or disease.
a. Morphological Changes
[0275] Apoptosis in many cell types is correlated with altered
morphological appearances. Examples of such alterations include,
but are not limited to, plasma membrane blebbing, cell shape
change, loss of substrate adhesion properties. Such changes are
readily detectable with a light microscope. Cells undergoing
apoptosis can also be detected by fragmentation and disintegration
of chromosomes. These changes can be detected using light
microscopy and/or DNA or chromatin specific dyes.
b. Altered Membrane Permeability
[0276] Often the membranes of cells undergoing apoptosis become
increasingly permeable. This change in membrane properties can be
readily detected using vital dyes (e.g., propidium iodide and
trypan blue). Dyes can be used to detect the presence of necrotic
cells. For example, certain methods utilize a green-fluorescent
LIVE/DEAD.TM. Cytotoxicity Kit #2, available from Molecular Probes.
The dye specifically reacts with cellular amine groups. In necrotic
cells, the entire free amine content is available to react with the
dye, thus resulting in intense fluorescent staining. In contrast,
only the cell-surface amines of viable cells are available to react
with the dye. Hence, the fluorescence intensity for viable cells is
reduced significantly relative to necrotic cells (see, e. g.,
Haugland, 1996 Handbook of Fluorescent Probes and Research
Chemicals, 6th ed., Molecular Probes, OR).
c. Dysfunction of Mitochondrial Membrane Potential
[0277] Mitochondria provide direct and indirect biochemical
regulation of diverse cellular processes as the main energy source
in cells of higher organisms. These process include the electron
transport chain activity, which drives oxidative phosphorylation to
produce metabolic energy in the form of adenosine triphosphate
(i.e., ATP). Altered or defective mitochondrial activity can result
in mitochondrial collapse called the "permeability transition" or
mitochondrial permeability transition. Proper mitochondrial
functioning requires maintenance of the membrane potential
established across the membrane. Dissipation of the membrane
potential prevents ATP synthesis and thus halts or restricts the
production of a vital biochemical energy source.
[0278] Consequently, a variety of assays designed to assess
toxicity and cell death involve monitoring the effect of a test
agent on mitochondrial membrane potentials or on the mitochondrial
permeability transition. One approach is to utilize fluorescent
indicators (see, e.g., Haugland, 1996 Handbook of Fluorescent
Probes and Research Chemicals, 6th ed., Molecular Probes, Oreg.,
pp. 266-274 and 589-594). Various non-fluorescent probes can also
be utilized (see, e.g., Kamo et al. (1979) J. Membrane Biol.
49:105). Mitochondrial membrane potentials can also be determined
indirectly from mitochondrial membrane permeability (see, e.g.,
Quinn (1976) The Molecular Biology of Cell Membranes, University
Park Press, Baltimore, Md., pp. 200-217). Further guidance on
methods for conducting such assays is provided in PCT publication
WO 00/19200 to Dykens et al.
d. Caspase Activation
[0279] Apoptosis is the process of programmed cell death and
involves the activation of a genetic program when cells are no
longer needed or have become seriously damaged. Apoptosis involves
a cascade of biochemical events and is under the regulation of a
number of different genes. One group of genes act as effectors of
apoptosis and are referred to as the interleukin-1.beta. converting
enzyme (ICE) family of genes. These genes encode a family of
cysteine proteases whose activity is increased in apoptosis. The
ICE family of proteases is generically referred to as caspase
enzymes. The "C" in the name reflects the fact that the enzymes are
cysteine proteases, while "Caspase" refers to the ability of these
enzymes to cleave after aspartic acid residues.
[0280] Consequently, some assays for apoptosis are based upon the
observation that caspases are induced during apoptosis. Induction
of these enzymes can be detected by monitoring the cleavage of
specifically-recognized substrates for these enzymes. A number of
naturally occurring and synthetic protein substrates are known
(see, e.g., Ellerby et al. (1997) J. Neurosci. 17:6165; Kluck, et
al. (1997) Science 275:1132; Nicholson et al. (1995) Nature 376:37;
and Rosen and Casciola- Rosen (1997) J. Cell Biochem. 64:50).
Methods for preparing a number of different substrates that can be
utilized in these assays are described in U.S. Pat. No. 5,976,822.
This patent also describes assays that can be conducted using whole
cells that are amendable to certain of the microfluidic devices
described herein. Other methods using FRET techniques are discussed
in Mahajan, et al. (1999) Chem. Biol. 6:401-9; and Xu, et al.
(1998) Nucl. Acids. Res. 26:2034-5.
e. Cytochrome C Release
[0281] In healthy cells, the inner mitochondrial membrane is
impermeable to macromolecules. Thus, one indicator of cell
apoptosis is the release or leakage of cytochrome C from the
mitochondria. Detection of cytochrome C can be performed using
spectroscopic methods because of the inherent absorption properties
of the protein. Thus, one detection option with the present devices
is to place the cells within a holding space and monitor absorbance
at a characteristic absorption wavelength for cytochrome C.
Alternatively, the protein can be detected using standard
immunological methods (e.g., ELISA assays) with an antibody that
specifically binds to cytochrome C (see, e.g., Liu et al. (1996)
Cell 86:147).
f. Assays for Cell Lysis
[0282] The final stage of cell death is typically lysis of the
cell. When cells die they typically release a mixture of chemicals,
including nucleotides, and a variety of other substances (e.g.,
proteins and carbohydrates) into their surroundings. Some of the
substances released include ADP and ATP, as well as the enzyme
adenylate cyclase, which catalyzes the conversion of ADP to ATP in
the presence of excess ADP. Thus, certain assays involve providing
sufficient ADP in the assay medium to drive the equilibrium towards
the generation of ATP which can subsequently be detected via a
number of different means. One such approach is to utilize a
luciferin/luciferase system that is well known to those of ordinary
skill in the art in which the enzyme luciferase utilizes ATP and
the substrate luciferin to generate a photometrically detectable
signal. Further details regarding certain cell lysis assays that
can be performed are set forth in PCT publication WO 00/70082.
g. Ischemic Model Systems
[0283] Methods for assaying whether a compound can confer
protective neurological effects against ischemia and stroke are
discussed by Aarts, et al. (Science 298:846-850, 2002). In general,
this assay involves subjecting rats to a middle cerebral artery
occlusion (MCAO) for a relatively short period of time (e.g., about
90 minutes). MCAO can be induced using various methods, including
an intraluminal suture method (see, e.g., Longa, E. Z. et al.
(1989) Stroke 20:84; and Belayev, L., et al. (1996) Stroke
27:1616). A composition containing the putative inhibitor is
introduced into the rat using conventional methods (e.g., via
intravenous injection). To evaluate the compositions prophylactic
effect, the composition is administered before performing MCAO. If
the compound is to be evaluated for its ability to mitigate against
an ischemic event that has already occurred, the composition with
the compound is introduced after MCAO has been initiated. The
extent of cerebral infarction is then evaluated using various
measures of neurological function. Examples of such measures
include the postural reflex test (Bederson, J. B. et al. (1986)
Stroke 17:472) and the forelimb placing test (De Ryck, M. et al.
(1989) Stroke 20:1383). Methods are also described in Aarts et al
assessing the effects of NMDA-induced excitotoxicity using in vitro
assays.
h. MTT Cytotoxicity Assay
[0284] The MTT assay is another assay which has been widely used to
assess cytotoxicity in neuronal cells. The cellular toxicity can
assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromide (MTT) assay (Trevigen, Gaithersburg, Md.)
following the recommendations of the manufacturer.
i. Trypan Blue Cell Viability Measurement
[0285] Cell viability can be measured using the trypan blue
exclusion method (Yao et al., Brain Res., 889, 181-190 (2001)).
j. Determination of Cellular ATP Levels
[0286] Cellular ATP Levels can be indicative of cell viability.
Cellular ATP concentrations can be measured using the
ATPLite-M.RTM. luminescence assay (Packard BioSciences Co.). For
example, in this assay, cells typically are cultured on black
96-well ViewPlate.RTM. and the ATP concentrations are measured on a
TopCount NXT.RTM. counter (Packard BioSciences Co.) following the
recommendations of the manufacturer.
vi) Gastrointestinal Absorption
[0287] The compounds or drugs according to the invention can be
further analyzed, tested or validated for their ability to be
absorbed by the gut and/or intestine if so desired.
[0288] Intestinal permeability and transport of a drug candidate
may be estimated using a variety of in vitro, in situ, as well as
in vivo models (Balimane et al. (2000) J Pharmacol Toxicol Methods
44:385-401; Hidalgo I. (2001) Curr Top Med Chem 1:385-401,
Hillgreen K, Kato A and Borchardt R. (1995) 15:83-109).
[0289] For instance, parallel artificial membrane permeability
(PAMPA) assay and cell-based systems such as Caco-2 and
Mardin-Darby canine kidney (MDCK) cells are the most frequently
used in vitro models. The PAMPA model consists of a hydrophobic
filter material coated with a mixture of lecithin/phospholipids
dissolved in an inert organic solvent creating an artificial lipid
membrane barrier that mimics the intestinal epithelium. Caco-2
cells, a human colon adenocarcinoma, undergo spontaneous
enterocytic differentiation in culture and become polarized cells
with well-established tight junctions, resembling intestinal
epithelium in humans. Caco-2 cell model has been the most popular
and the most extensively characterized cell-based model in
examining the permeability of drugs in, both the pharmaceutical
industries and academia. Alternatively, MDCK cells which also
develop tight junctions and form monolayers of polarized cells are
used.
[0290] An in situ study such as an intestinal perfusion could also
be performed to assess drug absorption. Isolated intestinal
segments comprise the absorptive cells and the underlying muscle
layers. As it is commonly used, this technique only allows sampling
from the mucosal side; drug disappearance is assumed to be equal to
drug absorption. Typically, a whole animal absorption study
(pharmacokinetic study) will be performed in parallel with the in
vitro and/or in situ studies to assess intestinal permeability. In
general, drug absorption in animals is believed to be a good
predictor of absorption in humans.
vii) Gastrointestinal Toxicity
[0291] The compounds or drugs according to the invention can be
further analyzed, tested or validated for gastrointestinal (GI)
toxicity. Gastrointestinal toxicity of a compound in vivo can be
reliably established through the implementation of a standard
battery of general toxicological assessments. Generally, regulatory
test guidelines from the EU, OECD, ICH, FDA and JMOHW are used as
reference material for the preparation of study protocols for such
assessments. In North America, the toxicological assessments will
generally be carried out in compliance with the United States Food
and Drug Administration Title 21 Code of Federal Regulations Part
58, Good Laboratory Practice for Non-clinical studies issued on
Dec. 22, 1978, Federal Register plus subsequent amendments.
[0292] Within the context of such a non-clinical assessment of the
toxicity of a particular compound, GI toxicity may specifically be
assessed through the monitoring of body weight gain, the gross
examination of materials emitted by the test subject (specifically
vomitus and feces) and the monitoring of food/water consumption
(appetence). Furthermore, upon termination of a non-clinical
toxicological assessment, the retention and processing of GI tract
tissues from the test subject(s) to the slide stage, followed by
histopathological examination of said tissues by a trained
pathologist, is a useful tool, complementary to the aforementioned
"in-life" observations.
viii) Crossing of the Blood Brain Barrier (BBB)
[0293] The blood-brain barrier (BBB) is a very specialized barrier
system of endothelial cells that separates the blood from the
underlying brain cells, providing protection to brain cells and
preserving brain homeostasis. The brain endothelium has a complex
arrangement of tight junctions between the cells that restrict the
passage of molecules. Typically the BBB is permeable to small and
lipophilic molecules, but larger molecules are generally not
transported across unless there is an active transport system
available. Thus this is one of the stumbling blocks for drug
delivery. An additional problem is the very effective drug efflux
systems (P-glycoprotein), which pump the drug back out of
cells.
[0294] The compounds according to the invention can be further
analyzed, tested or validated for their ability to cross the BBB is
so desired. Many in-vitro, in-vivo and in-silico methods may be
employed during drug development to mimic the BBB (Lohmann et al.
(2002) Predicting blood-brain barrier permeability of drugs:
evaluation of different in vitro assays. J Drug Target 10:263-276;
Nicolazzo et al. (2006) Methods to assess drug permeability across
the blood-brain barrier. J Pharm Pharmacol 58:281-293). In-vitro
models include primary endothelial cell culture and immortalized
cell lines such as Caco-2, BMEC, MDCK. These cells are useful as a
screening method and can appropriately rank compounds in order of
BBB permeability. In vivo models such as the internal carotid
artery single injection or perfusion, intravenous bolus injection,
brain efflux index and intracerebral microdialysis provide more
accurate information regarding brain uptake, and these can be
complemented with novel imaging techniques (such as magnetic
resonance imaging and positron emission tomography), although such
methods are not suited to high-throughput permeability
assessment.
[0295] ix) Brain and CSF Level
[0296] The brain and/or cerebrospinal fluid (CSF) levels of the
compounds or drugs according to the invention can be assessed,
measured or estimated using various models methods, and assays (see
Potchoiba M J, and Nocerini, M R (2004) DMD 32:1190-1198;
Orlowska-Madjack M. (2004) Acta Neurobiol Exp 64: 177-188; and
Hocht, C, Opezza, J A and Taira, C A (2004) Curr Drug Discov
Technol 1:269-85)
[0297] One of the most common techniques is probably a brain
sampling after a whole animal absorption study (pharmacokinetic).
For instance, pharmacokinetics (PK) profiles of the compound of the
invention could be investigated using typical nonclinical PK
studies in mice. Briefly, at different time-points following
intravenous, subcutaneous and oral compound administrations, brain,
CSF and plasma samples are collected. The brain, CSF and plasma
samples are then analyzed by LC/MS to determine the
concentration-time profiles of the compound.
[0298] Alternatives such as brain dialysis or distribution of a
radiolabelled compound with or without autoradioluminography could
also be used. A typical example is a tissue distribution study to
assess the time course elimination of radioactivity from tissues
following the administration of a known quantity of radiolabeled
compound, the percentage of the original dose transported in the
brain or CSF can be determined. Furthermore, autoradioluminography
of cryosections containing brain tissues with a wide range of
radioactivity concentrations can be readily quantified to determine
brain levels of a drug.
[0299] Alternatively, microdialysis offers a way to remove drugs
from the brain. The principle of microdialysis is based on the
diffusion of molecules through small-diameter pores of a semi
permeable membrane tubing connected to a probe that is implanted
into a defined brain area. The probe is connected to a perfusion
pump and perfused with a liquid, which equilibrates with the fluid
outside the tube by diffusion in both directions. A quantitative
analysis of drug in the fraction-collected microdialysates reflects
their concentration in the fluid.
[0300] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures, embodiments, claims, and
examples described herein. Such equivalents are considered to be
within the scope of this invention and covered by the claims
appended hereto. The contents of all references, issued patents,
and published patent applications cited throughout this application
are hereby incorporated by reference. The invention is further
illustrated by the following examples, which should not be
construed as further limiting.
EXAMPLES
[0301] The Examples set forth herein below provide exemplary
syntheses of certain representative compounds of the invention.
Also provided are exemplary methods for assaying the compounds of
the invention for in vitro stability, microsomes metabolism and
mouse bioavailability.
[0302] Unless otherwise indicated, all numbers expressing
quantities of ingredients, reaction conditions, concentrations,
properties, and so forth used in the specification and claims are
to be understood as being modified in all instances by the term
"about" At the very least, each numerical parameter should at least
be construed in light of the number of reported significant digits
and by applying ordinary rounding techniques. Accordingly, unless
indicated to the contrary, the numerical parameters set forth in
the present specification and attached claims are approximations
that may vary depending upon the properties sought to be obtained.
Notwithstanding that the numerical ranges and parameters setting
forth the broad scope of the embodiments are approximations, the
numerical values set forth in the specific examples are reported as
precisely as possible. Any numerical value, however, inherently
contain certain errors resulting from variations in experiments,
testing measurements, statistical analyses and such.
[0303] The present invention also relates to novel compounds and
the synthesis thereof. The following detailed examples describe how
to prepare the various compounds and/or perform the various
processes of the invention and are to be construed as merely
illustrative, and not limitations of the preceding disclosure in
any way whatsoever. Those skilled in the art will promptly
recognize appropriate variations from the procedures both as to
reactants and as to reaction conditions and techniques. In some
cases, the compounds may be commercially available.
Example 1-A: Chemical Synthesis of Amino Acid Prodrugs
[0304] Accordingly, the following examples are presented to
illustrate how some amino acid prodrugs according to the invention
compounds may be prepared.
Preparation of N-hydroxysuccinimide Ester
##STR00129##
[0306] To a stirred solution of a N-Boc-protected amino acid or a
carboxylic acid (10 mmol) in CH.sub.2Cl.sub.2 (100 mL) was added
HBTU (N,N,N',N'-Tetramethyl-O-(1H-benzotriazol-1-yl)uronium
hexafluorophosphate, 4.17 g, 11 mmol) followed by addition of
triethylamine (1.53 mL, 11 mmol) and N-hydroxysuccinimide (NHS,
1.26 g, 11 mmol). The reaction mixture was stirred at room
temperature for 4 h, and then diluted with HCl (1 N) and EtOAc
(ethyl acetate). The organic layer was isolated, dried over
Na.sub.2SO.sub.4, and concentrated. The residual material was
purified by flash chromatography on silica gel using hexanes-EtOAc
as eluent to afford the corresponding N-hydroxysuccinimide ester in
good yield (about 70 to 88%).
General Procedures for the Preparation of Amino Acid Prodrugs of
3-amino-1-propanesulfonic Acid (Procedures A to D):
[0307] Procedures A to D were used in different combinations, to
produce exemplary compounds of the invention. Results for the
preparation of Compounds A to Y using these procedures are
summarized in Table 2 below.
Procedure A:
##STR00130##
[0309] A solution of the N-hydroxysuccinimide ester of a
N-Boc-protected amino acid or a carboxylic acid (48 mmol, 1.2 eq)
in acetonitrile or acetone (50 mL) was added slowly to a solution
of 3APS, 3-amino-1-propanesulfonic acid, 40 mmol, 1 eq in 2 N NaOH
(sodium hydroxide, 23 mL, 1.2 eq). The reaction mixture was stirred
at room temperature overnight. The mixture was evaporated to
dryness. The residual material was stirred with Et.sub.2O (diethyl
ether, 150 mL) at reflux for 1 h. After the mixture was cooled to
room temperature, the solid material was filtered and dried in
vacuo, and further purified according to one of the following
work-up procedures: [0310] (i) The solid material was dissolved in
water (25 mL). The solution was passed through a Dowex.TM.
Marathon.TM. C ion-exchange column (strongly acidic, 110 g (5 eq),
pre-washed). The strong acidic fractions were combined and treated
with concentrated HCl (10 mL). The mixture was stirred at
50.degree. C. for 30 minutes, and then was concentrated to dryness.
The residual material was co-evaporated with EtOH (ethanol) to
completely remove water. EtOH (100 mL) was added to the residue.
The mixture was stirred at reflux for 1 h, and then cooled to room
temperature. The solid material was collected by filtration. The
solid material was dissolved in water (10 mL). The solution was
added drop wise to EtOH (100 mL). The product slowly crystallized.
The suspension was stirred at room temperature for 30 minutes. The
solid material was collected by filtration and it was dried in a
vacuum oven (60.degree. C.). [0311] (ii) The solid material was
dissolved in water (25 mL). The solution was passed through a
Dowex.TM. Marathon.TM. C ion exchange column (strongly acidic, 110
g (5 eq), pre-washed). The strong acidic fractions were combined
and evaporated under reduced pressure. The residue was purified
using reverse-phase flash chromatography (Biotage.TM. SP-1, C18
column). For ester-containing compound, the final product was
obtained after removal of the solvent from the corresponding
fractions; otherwise go to (iii). [0312] (iii) The residual
material from step (ii) above was stirred with 4N HCl (3 mL) at
50.degree. C. for 1 h. A white solid precipitate appeared. After
the mixture was cooled to room temperature, the solid material was
collected by filtration, washed, and dried in vacuo, to provide the
final product.
Procedure B:
[0313] To a stirred solution of a N-hydroxysuccinimide ester (3
mmol) in a mixture of H.sub.2O/tetrahydrofuran/CH.sub.3CN (10/10/10
mL) was added a solution of SAPS (as sodium salt) (3.3 mmol) in
water (5 mL) followed by addition of 1M solution of potassium
carbonate (3 mL). The reaction mixture was stirred for 2 h,
followed by addition of EtOAc. The aqueous layer was isolated and
concentrated to a residue. The residual material was purified by
silica gel column using CH.sub.2Cl.sub.2-MeOH (80-20) as eluent to
give the corresponding N-Boc-protected product. The purified
N-Boc-protected product was dissolved in dichloromethane
(CH.sub.2Cl.sub.2, 10 mL) followed by addition of TFA
(trifluoroacetic acid, 5 mL). The reaction mixture was stirred for
2 h, and then concentrated under reduced pressure. The residual
solid material was suspended in a minimum amount of ethanol and the
mixture was stirred for 1 h under reflux. The mixture was cooled to
room temperature. The solid material was collected by filtration,
washed with ethanol, and dried under high vacuum to afford the
final compound.
Procedure C:
[0314] To the purified product containing benzyl ether protection
group from procedure A or B (3.5 mmol) in 2N HCl (500 mL) and MeOH
(500 mL) was added 10% Pd/C (2.15 g). The mixture was stirred under
hydrogen (1 atm) overnight. The suspension was filtered
(Celite.TM.. The filter cake was washed with water (2.times.25 mL).
The filtrate and the washing were combined and evaporated under
reduced pressure. The residual material was purified by
reverse-phase HPLC (C18 column, 0-15% acetonitrile/water). The
fractions containing the desired compound were combined and
lyophilized, to give the final product.
Procedure D:
[0315] This procedure is used to produce prodrugs of Formulae I to
VI having more than one amino acid coupled to 3APS. Step (i) or
(ii) is repeated as necessary to obtain the desired compound.
##STR00131## [0316] (i) The product from Procedures A, B, or C is
further reacted with another N-hydroxysuccinimide ester following
Procedure A(i). [0317] (ii) The product from Procedures A, B, or C
was further reacted with another N-hydroxysuccinimide ester
following Procedure B.
TABLE-US-00006 [0317] TABLE 5 Synthesis and characterization of
exemplary amino acid prodrugs according to the invention Synthetic
NMR (ppm; 1H 500 MHz; 13C 125 MHz) ID Procedure MS (electrospray
ionization) A1 A(i) 1H NMR (D2O) .delta. 1.55-1.61 (m, 2H),
2.40-2.48 (m, 2H), 2.92-3.01 (m, 2H), 3.04-3.14 (m, 2H), 3.95-3.98
(m, 1H), 7.11 (d, J = 6.8 Hz, 2H), 7.197.27 (m, 3H); 13C NMR (D2O)
.delta. 23.76, 37.02, 38.21, 48.36, 54.79, 128.19, 129.33, 129.42,
134.01, 168.94; m/z 285 (M - 1). A2 A(i) 1H NMR (D2O) .delta.
0.87-0.90 (m, 6H), 1.83 (qt, J = 7.2 Hz, 2H), 2.02-2.09 (m, 1H),
2.79 (t, J = 7.8 Hz, 2H), 3.20-3.29 (m, 2H), 3.60 (d, J = 6.3 Hz,
2H); 13C NMR (D2O) .delta. 17.20, 17.77, 24.11, 30.00, 38.29,
48.63, 58.96, 169.35; m/z 237 (M - 1). A3 A(i) 1H NMR (D2O) .delta.
1.82 (qt, J = 7.2 Hz, 2H), 1.90-1.95 (m, 3H), 2.28-2.33 (m, 1H),
2.78 (t, J = 7.8 Hz, 2H), 3.22-3.33 (m, 4H), 4.21 (t, J = 7.1 Hz,
2H); 13C NMR (D2O) .delta. 23.95, 24.07, 29.85, 38.49, 46.57,
48.53, 60.00, 169.64; m/z 235 (M - 1). A4 A(ii) 1H NMR (D2O)
.delta. 1.30 (qt, J = 8.1 Hz, 2H), 1.57 (qt, J = 7.8 Hz, 2H),
1.75-1.85 (m, 4H), 2.77-280 (m, 2H), 2.87 (t, J = 7.8 Hz, 2H), 3.17
(qt, J = 6.7 Hz, 1H), 3.31 (qt, J = 6.8 Hz, 1H), 3.83 (t, J = 6.6
Hz, 1H); 13C NMR (D2O) .delta. 21.47, 24.12, 30.49, 38.30, 39.18,
48.63, 53.28, 169.66; m/z 266 (M - 1). A5 B 1H NMR (DMSO-d6)
.delta. 0.81 (d, J = 7.3 Hz, 3H), 7.84 (d, J = 7.3 Hz, 3H), 1.5 (m,
1H), 1.60 (m, 2H), 1.82 (m, 2H), 2.80 (m, 2H), 3.20-3.30 (m, 2H),
3.82 (t, J = 7.3 Hz, 1H); 13C NMR (DMSO-d6) .delta. 21.48, 21.78,
24.17, 38.42, 40.08, 48.66, 52.35, 170.53; m/z 251 (M - 1). A6 A(i)
1H NMR (D2O) .delta. 1.84 (m, 2H), 1.99 (s, 3H), 2.04 (m, 2H), 2.47
(m, 2H), 2.80 (m, 2H), 3.24 (t, J = 6.6 Hz, 2H), 3.94 (t, J = 6.6
Hz, 2H); 13C NMR (D2O) .delta. 14.18, 24.07, 28.44, 30.09, 38.41,
48.61, 52.66, 169.46; m/z 269 (M - 1). A7 B and C 1H NMR (D2O)
.delta. 1.81 (m, 2H), 2.80 (m, 2H), 3.23 (m, 2H), 3.80 (m, 2H),
3.97 (t, J = 5.0 Hz, 1H); 13C NMR (D2O) .delta. 24.10, 38.39,
48.55, 54.85, 60.44, 167.97; m/z 225 (M - 1). A8 A(i) 1H NMR (D2O)
.delta. 3.90 (q, 1H, J = 7 Hz), 3.23 (t, 2H, J = 7 Hz), 2.78 (m,
2H), 1.82 (m, 2H), 1.38 (d, 3H, J = 7 Hz); 13C NMR (D2O) .delta.
170.90, 49.30, 48.55, 38.28, 24.10, 16.65; m/z 209 (M - 1). A9 A(i)
1H NMR (D2O) .delta. 3.90 (q, 1H, J = 7 Hz), 3.23 (t, 2H, J = 7
Hz), 2.78 (m, 2H), 1.82 (m, 2H), 1.38 (d, 3H, J = 7 Hz); 13C NMR
(D2O) .delta. 170.90, 49.30, 48.55, 38.28, 24.10, 16.65; m/z 209 (M
- 1). A10 B 1H NMR (D2O) .delta. 1.82 (m, 2H), 2.80 (m, 2H), 3.25
(m, 2H). 3.67 (s, 2H); 13C NMR (D2O) .delta. 24.13, 38.26, 40.57,
48.55, 167.08; m/z 195 (M - 1). A11 A(i) 1H NMR (D2O) .delta. 0.80
(t, 3H, J = 7.3 Hz), 0.86 (d, 3H, J = 6.8 Hz), 1.12 (m, 1H), 1.40
(m, 1H), 1.83 (m, 3H), 2.79 (m, 2H), 3.25 (m, 2H), 3.68 (d, 1H, J =
5.9 Hz); 13C NMR (D2O) .delta. 10.59, 14.22, 24.11, 24.37, 36.38,
38.29, 48.64, 58.00, 169.35; m/z 251 (M - 1). A12 A(i) 1H NMR (D2O)
.delta. 1.84 (m, 2H), 1.99 (s, 3H), 2.04 (m, 2H), 2.47 (m, 2H),
2.80 (m, 2H), 3.25 (t, J = 7.3 Hz, 2H), 3.94 (t, J = 6.6 Hz, 1H);
13C NMR (D2O) .delta. 14.18, 24.06, 28.42, 30.07, 38.41, 48.60,
52.66, 169.42; m/z 269 (M - 1). A13 A(i) 1H NMR (D2O) .delta. 1.70
(m, 2H), 2.64 (m, 2H), 3.15 (m, 1H), 3.22 (m, 3H), 4.06 (t, J = 6.3
Hz, 1H), 7.30 (s, 1H), 8.55 (d, J = 1.5 Hz, 1H); 13C NMR (D2O)
.delta. 23.94, 26.27, 38.36, 48.43, 52.59, 118.40, 126.36, 134.60,
167.96; m/z 275 (M - 1). A14 A(i) 1H NMR (D2O) .delta. 1.46 (s,
6H), 1.83 (m, 2H), 2.77 (m, 2H), 3.23 (t, J = 6.6 Hz, 2H); 13C NMR
(D2O) .delta. 23.44, 24.08, 38.54, 48.61, 57.21, 173.20; m/z 223 (M
- 1). A15 A(i) 1H NMR (D2O) .delta. 1.74 (m, 2H), 2.59 (m, 2H),
3.15 (m, 1H), 3.23 (m, 1H), 4.95 (s, 1H), 7.38 (m, 5H); 13C NMR
(D2O) .delta. 24.00, 38.35, 48.38, 56.84, 128.05, 129.87, 130.52,
132.46, 168.90; m/z 271 (M - 1). A16 A(i) 1H NMR (D2O) .delta. 1.49
(m, 2H), 2.34 (m, 2H), 2.98 (m, 2H), 3.21 (m, 2H), 4.01 (m, 1H),
7.05 (t, 1H, J = 7.3 Hz), 7.14 (m, 2H), 7.39 (d, 1H, J = 8.3 Hz),
7.47 (m, 1H); 13C NMR (D2O) .delta. 23.60, 27.14, 38.32, 48.16,
54.12, 106.83, 112.32, 118.23, 119.70, 122.35, 125.18, 126.63,
136.37, 139.57; m/z 324 (M - 1). A17 A(iii) 1H NMR (D2O) .delta.
1.66 (m, 2H), 2.58 (m, 2H), and 2.92 (m, 1H), 3.04 (m, 2H), 3.17
(m, 1H), then C 3.95 (t, 1H, J = 6.3 Hz), 6.77 (d, 2H, J 8.8 Hz),
7.02 (d, 2H, J = 8.3 Hz); 13C NMR (D2O) .delta. 23.91, 36.29,
38.25, 48.42, 54.95, 116.07, 125.88, 130.91, 155.29, 169.56; m/z
301 (M - 1). A18 B 1H NMR (D2O) .delta. 1.77 (m, 2H), 2.74 (m, 2H),
3.19 (, m2H), 3.75 (m, 2H), 4.05 (m, 1H), 4.42 & 4.65 (AB, J =
12.2 Hz, 2H), 7.26-7.33 (m, 5H); 13C NMR (D2O) .delta. 24.10,
38.43, 53.23, 67.39, 73.28, 128.63, 128.67, 128.96, 136.86, 167.55;
m/z 315 (M - 1). A19 A(ii) 1H NMR (D2O) .delta. 7.3-7.2 (m, 5H),
5.05 (s, 2H), 3.83 (t, J = 6.7 Hz, 1H), 3.21 (qn, J = 7 Hz, 1H),
3.08 (qn, J = 7 Hz, 1H), 2.78 (t, J = 7.8 Hz, 2H), 2.45 (t, J = 7
Hz, 2H), 2.05 (q, J = 7 Hz, 2H), 1.78 (m, 2H); m/z 357 (M - 1). A20
B 1H NMR (D2O) .delta. 1.78-1.85 (m, 4H), 2.24 (t, J = 7.5 Hz, 2H),
2.79 (m, 2H), 2.88 (t, J = 7.8 Hz, 2H), 3.18 (t, J = 7.0 Hz, 2H).
13C NMR (D2O) .delta. 23.21, 24.16, 32.70, 38.16, 38.98, 48.65,
175.06; m/z 223 (M - 1). A22 1H NMR (D2O) .delta. 1.84 (qn, 2H, J =
7 Hz), 2.78 (dd, 2H, J = 8.0, 6 Hz), 2.85 (ABX, 2H, J = 5.5, 7.3,
16.8 Hz), 3.24 (m, 2H), 3.61 (dd, 1H, J = 5.5, 7.3 Hz); 13C NMR
(D2O) .delta. 24.05, 35.42, 38.46, 48.53, 50.04, 169, 171. A28 1H
NMR (D2O) .delta. 0.8-0.9 (m, 12H), 1.81 (m, 1H), 1.88 (m, 1H),
2.09 (m, 2H), 2.77 (t, 2H, J = 8.0 Hz), 3.20 (t, 2H, J = 6.6 Hz),
3.73 (d, 1H, J = 6.1 Hz), 3.87 (d, 1H, J = 8.9 Hz); 13C NMR (D2O)
.delta. 16.93, 17.82, 18.36, 24.21, 29.77, 30.27, 38.08, 48.72,
58.42, 60.66, 169.45, 173.07
Example 1-B: Chemical Synthesis of Carbamate Prodrugs
[0318] Accordingly, the following examples are presented to
illustrate how some carbamate prodrugs according to the invention
compounds may be prepared.
General Synthetic Procedures
Procedure A:
Preparation of Compound C1 Sodium Salt
(3-(p-acetyloxybenzyloxycarbonyl)amino-1-propanesulfonic Acid
Sodium Salt)
##STR00132##
[0320] Step 1: Acetyl chloride (3.0 mL, 42 mmol, 1 eq.) was added
to a mixture of 4-hydroxybenzylalcohol (5.3 g, 42 mmol), sodium
hydroxide (1.7 g, 42 mmol, 1 eq.) and tetrabutylammonium hydrogen
sulfate (7 g, 0.5 eq.) in dioxane (100 mL). The reaction mixture
was stirred at room temperature for 4 hours and the solvent was
evaporated. The residue was dissolved in water and the aqueous
phase was extracted with EtOAc (3 times). Combined organic extracts
were washed with brine, dried and concentrated to give colorless
oil. Purification (flash chromatography; hexane/EtOAc, gradient
mode) provided the corresponding monoacetate (2.2 g, 32%).
[0321] Step 2: Anhydrous pyridine (1.1 mL, 13 mmol, 1 eq,) was
added drop wise to a stirred mixture of p-nitrophenyl chloroformate
(4.0 g, 20 mmol, 1.5 eq.) and the monoacetate (from step 1: 2.2 g,
13 mmol) in dry tetrahydrofuran (THF, 25 mL). A white precipitate
was formed. The reaction mixture was stirred at room temperature
for 1 hour. The solid material was removed by filtration, and
washed with THF. The filtrate and the washing were combined; and
the solvent was removed in vacuo. The residual material was
purified by flash chromatography (hexanes/EtOAc, 80/20) to provide
the corresponding carbonate (2.8 g, 62%).
[0322] Step 3: The carbonate prepared in the step 2 (2.2 g, 6.7
mmol, 2 eq.) was added to a mixture of 3-amino-1-propanesulfonic
acid sodium salt (538 mg, 3.32 mmol) and triethylamine (0.90 ml,
6.7 mmol, 2 eq.) in dry N,N-dimethylformamide (DMF, 10 mL). The
reaction mixture was stirred at room temperature overnight. Solvent
was removed by evaporation. The residue was partitioned between
EtOAc and water. The aqueous phase was washed twice with EtOAc, and
then lyophilized. HPLC purification (acetonitrile/water, 20/80 to
90/10) of the lyophilized residue provided the title compound (396
mg, 33%): .sup.1H NMR (500 MHz, D.sub.2O) .delta. ppm 1.83-1.89 (m,
2H), 1.98 (s, 3H), 2.84-2.87 (m, 2H), 3.19-3.21 (m, 2H), 5.01 (s,
2H), 7.03 (d, J=8.8 Hz, 2H), 7.32 (d, J=8.3 Hz, 2H).
Procedure B:
Preparation of Compound C6 sodium salt
(4-aza-7-methyl-15-phenyl-11,11-tetramethylene-6,8,14-trioxa-5,9,13-triox-
o-1-pentadecanesulfonic acid sodium salt)
##STR00133##
[0324] Step 1: 3,3-Tetramethyleneglutaric acid monobenzyl ester
(4.26 g; 15.4 mmol, prepared by heating overnight the cyclic
anhydride and benzyl alcohol in dioxane at 80.degree. C. in the
presence of triethylamine) and silver oxide (2.13 g; 9.22 mmol)
were added to a mixture of acetonitrile (40 mL) and water (20 mL).
The mixture was heated at 70.degree. C. for 3 h, and then cooled to
room temperature. The mixture was filtered through a pad of
Celite.TM.. The filtrate was evaporated to provide the crude silver
carboxylate (2.19 g, 37%) which was used in the next step without
further purification.
[0325] Step 2: A mixture of the silver carboxylate (2.19 g, 5.71
mmol; from step 1) and the carbamating reagent (1.00 g; 2.95 mmol;
for preparation, see in Procedure E), in dry toluene (100 mL) was
heated at 50.degree. C. overnight. The mixture was filtered through
a pad of Celite.TM. and the filtrate was evaporated to provide a
solid residue, which was purified by flash chromatography using
hexane/EtOAc (80/20), giving the desired intermediate product
(0.915 g, 64%).
[0326] Step 3: To a solution of the intermediate product from step
2 (0.915 g; 1.88 mmol) in dry DMF (5 mL) was added
3-amino-1-propanesulfonic acid sodium salt (300 mg; 1.85 mmol). The
mixture was stirred at room temperature overnight. Solvent was
removed by evaporation. The residual material was purified by
Prep-HPLC to furnish, after lyophilization, the title compound (632
mg, 66%): .sup.1H NMR (CD.sub.3OD, 500 MHz) .delta. 1.39 (d, J=5.9
Hz, 3H), 1.64-1.59 (m, 8 H), 1.97-1.91 (m, 2H), 2.49 (qAB, J=15.1
Hz, 2H), 2.57 (qAB, J=15.1 Hz, 2H), 2.82-2.79 (m, 2H), 5.10 (s,
2H), 3.26-3.14 (m, 2H), 6.74 (q, J=5.9 Hz, 1H), 7.38-7.29 (m,
5H).
[0327] Other compounds prepared according to this procedure
(Procedure B) were purified either by precipitation using methanol
and ether (protocol (b)), or by preparative HPLC using
acetonitrile/water (10/90 to 90/10) over 40 minutes at 50 mL/min
(protocol (a)), or by normal phase flash chromatography (protocol
(c)).
Procedure C:
Preparation of Compound C2 sodium salt
(4-aza-12-carboxy-6,8-dioxa-5,9-dioxo-7-methyl-11,11-tetramethylene-1-dod-
ecanesulfonic acid sodium salt)
##STR00134##
[0329] The corresponding benzylester of the title compound (344 mg;
0.678 mmol) in methanol (5 mL) was hydrogenolyzed in the presence
of Pd/C 10% (100 mg) at 40-45 psi for 1 h. The mixture was filtered
(Celite.TM. and the filtrate was evaporated to dryness. The
residual material was dissolved in water and the aqueous solution
was lyophilized, giving the title compound (242 mg, 86%): .sup.1H
NMR (CD.sub.3OD, 500 MHz) .delta. 1.43 (d, J=5.4 Hz, 3H), 1.66-1.63
(m, 8H), 1.98-1.92 (m, 2H), 2.49 (qAB, J=15.6 Hz, 2H), 2.55 (qAB,
J=15.1 Hz, 2H), 2.83-2.80 (m, 2H), 3.24-3.21(m, 2H), 6.77 (q, J=5.4
Hz, 1H), 7.22 (t, J=5.4, N--H).
[0330] Procedure D:
Preparation of Compound C19 sodium salt
(4-aza-7-methyl-6,8,-dioxa-5,9,-dioxo-1-decanesulfonic acid sodium
salt)
##STR00135##
[0332] Step 1: 1-Chloroethylchloroformate (7.8 ml, 72 mmol, 1 eq.)
was added to an ice-cold solution of p-nitrophenol (10 g, 72 mmol)
in chloroform (100 mL), followed by drop wise addition of pyridine
(8.8 ml, 108 mmol, 1.5 eq.) over a period of 20 min. The mixture
was stirred in the ice-cold bath for 15 min, and then at room
temperature overnight. The reaction mixture was sequentially washed
with water, 1 N hydrochloric acid, water, 1 N sodium hydroxide,
water, and brine. The organic phase was dried over
Na.sub.2SO.sub.4, and concentrated to give yellow oil which, upon
standing, crystallized to afford the corresponding chloroethyl
carbonate (15.5 g, 88%).
[0333] Step 2: To a solution of the chloroethyl carbonate obtained
from step 1 (6.2 g, 25 mmol) in acetic acid (150 mL) was added
mercuric acetate (9.6 g, 30 mmol, 1.2 eq.). The mixture was stirred
at room temperature overnight. Solvent was evaporated. The residual
material was transferred into ether and washed with a saturated
aqueous solution of NaHCO.sub.3. The ether layer was dried over
MgSO.sub.4 and concentrated to give thick yellow oil. Purification
of the oil by flash chromatography (hexane/EtOAc, 95/5) gave the
corresponding acetyloxyethyl carbonate (6.3 g, 94%) as colorless
oil.
[0334] Step 3: The acetyloxyethyl carbonate obtained from step 2
(1.2 g, 4.3 mmol, 1.1 eq.) was added to a solution of
3-amino-1-propanesulfonic acid sodium salt (0.63 g, 3.9 mmol) in
DMF (10 mL). The yellow solution was stirred at room temperature
overnight (color disappeared at this point). Solvent was
evaporated. The residue was triturated several times with ether and
turned to a solid. The solid material was collected by filtration
to give the title compound (840 mg, 74%): .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. 1.42 (d, J=5.4 Hz, 3H), 1.92-1.98 (m, 2H), 2.02
(s, 3H), 2.80-2.83 (m, 2H), 3.20-3.24 (m, 2H), 6.73 (q , J=5.4 Hz,
1H)
[0335] Other compounds prepared according to this procedure
(Procedure D) were purified either by extraction from EtOAc/water
followed by lyophilization of the aqueous phase, or reverse-phase
HPLC purification using acetonitrile/water (10/90 to 90/10) in 40
minutes at 50 mL/min, or trituration/precipitation with ether.
Procedure E:
Preparation of Compound C16 sodium salt
(4-aza-7-methyl-6,8,-dioxa-5,9,-dioxo-9-phenyl-1-nonanesulfonic
acid sodium salt)
##STR00136##
[0337] Step 1: Sodium iodide (14 g, 92 mmol, 3 eq.) was added to a
mixture of the chloroethyl carbonate (7.5 g, 31 mmol; for
preparation, see in Procedure D), and grinded calcium chloride (10
g, 92 mmol, 3 eq.) in acetonitrile (100 mL). The reaction mixture
was stirred at 40 .degree. C. for 4 days, followed by filtration
through a Celite.TM. pad. The filtrate was concentrated to give a
red gummy residue. Purification by flash chromatography using
EtOAc/hexane in a gradient mode provided the corresponding
iodoethyl carbonate (6 g, 59%) as pale yellow oil.
[0338] Step 2: Silver benzoate (5.5 g, 24 mmol, 2 eq.) was added to
a solution of the above-obtained iodoethyl carbonate (4 g, 12 mmol)
in toluene (50 mL). The reaction mixture was stirred at 55.degree.
C. overnight. The reaction mixture was filtered through a
Celite.TM. pad and washed with toluene. The filtrate was
concentrated to give brown oil. Two repeated purifications by flash
chromatography using hexane/EtOAc (90/10) provided the
corresponding benzoate (0.98 g, 25%) in high purity.
[0339] Step 3: The above-obtained benzoate (0.98 g, 2.9 mmol, 1.1
eq., from step 2) was added to a solution of
3-amino-1-propanesulfonic acid sodium salt (0.43 g, 2.7 mmol) in
DMF (10 mL). The yellow solution was stirred at room temperature
overnight. Solvent was evaporated and the residue was dissolved in
water. The aqueous solution was extracted several times with EtOAc.
The aqueous phase was lyophilized to give a residue, which was
purified by preparative HPLC (acetonitrile/water; 10/90 to 90/10,
in 40 minutes at 50 mL/min), giving the title compound (256 mg):
.sup.1H NMR (500 MHz, D.sub.2O) .delta. 1.48 (d, J=5.4 Hz, 3H),
1.76-1.82 (m, 2H), 2.76-2.79 (m, 2H), 3.08-3.14 (m, 2H), 6.83 (q,
J=5.4 Hz, 1H), 7.39-7.42 (m, 2H), 7.55-7.58 (m, 1H), 7.89-7.91 (m,
2H).
Procedure F:
Preparation of Compound C26 sodium salt
(3-({[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy]carbonyl}amino)-1-propanesu-
lfonic acid sodium salt)
##STR00137##
[0341] A mixture of the sodium salt of 3-amino-1-propanesulfonic
acid (532 mg; 3.30 mmol) and the carbonate (1.10 g; 3.73 mmol;
ref., J. Med. Chem., 1996, 39, 480-486) in dry DMF (10 mL) was
stirred at room temperature overnight. Solvent was removed in
vacuo. To the residual material was added methanol (10 mL),
followed by the addition of ether (75 mL). The solid formed was
collected by filtration and dried overnight. Again the solid was
dissolved in methanol (10 mL) and precipitated with ether (50 mL).
The solid material was purified by preparative HPLC to provide the
title compound (260 mg, 25%) as a white lyophilized solid: .sup.1H
NMR (CD.sub.2OD, 500 MHz) .delta. 1.98-1.92 (m, 2H), 2.17 (s, 3H),
2.90-2.79 m, 2H), 3.22 (t, J=6.8 Hz, 2H), 4.86 (s, 2H).
TABLE-US-00007 TABLE 6 Synthesis and characterization of exemplary
carbamate prodrugs according to the invention Synthetic Purifying
m/z (ES.sup.-) ID procedure protocol* (M - H, or M -
Na).sup..dagger. C1 A (a) 330.0 C2 C (d) 394.0 C3 B, C (a) 408.5 C4
C (d) 326.1 C5 B (a) 416.0 C6 B (a) 484.0 C7 B (a) 458.3 C8 C (d)
368.5 C9 C (d) 354.0 C10 B (a) 444.1 C11 C (d) 340.1 C12 B (b) and
(a) 430.2 C13 B (b) and (a) 378.0 C14 B (b) and (a) 372.0 C15 D (a)
310.2 C16 E (a) 330.2 C17 D (a) 336.2 C18 D (b) 296.2 C19 D (b)
268.1 C20 D (a) 378.1 C21 D (a) 310.1 C22 D (a) 296.1 C23 D (a)
338.1 C24 D (a) 310.0 C25 E (b) 253.9 C26 F (b) and (a) 294.0 *(a),
HPLC; (b), precipitation; (c), flash chromatography; (d),
filtration; (e), extraction, .sup..dagger.the compounds were
synthesized as acid form, or as sodium salt form.
Example 1-C: Chemical Synthesis of Non-Amino Acid Amide
Prodrugs
[0342] Accordingly, the following examples are presented to
illustrate how some non-amino acid amide prodrugs according to the
invention compounds may be prepared.
Procedure A:
Preparation of Compound B3 sodium salt
(3,3-dimethyl-5-oxo-5-[(3-sulfopropyl)amino]pentanoic acid sodium
salt)
##STR00138##
[0344] A mixture of the 3,3-dimethylglutaric anhydride (1.0 g; 7.0
mmol) and 3-amino-1-propanesulfonic acid sodium salt (0.950 g; 5.86
mmol) in dry DMF (20 mL) was stirred at 50.degree. C. for 2 days.
Solvent was evaporated. To the residual material was added methanol
(.about.10 mL) followed by the addition of ether (.about.50 mL) to
cause precipitation. The precipitate formed was collected by
filtration and then dissolved in water and lyophilized to provide
the title compounds (1.33 g, 75%) as a powder: .sup.1H NMR
(D.sub.2O, 500 MHz) .delta. 0.94 (s, 6H), 1.82-1.77 (m, 2H), 2.14
(s, 3H), 2.23 (s, 3H), 2.79-2.76 (m, 2H), 3.16 (t, J=6.8 Hz,
2H).
[0345] Other compounds prepared in the above procedure (Procedure
A, see Table 7) were purified either by methanol-ether
precipitation (Purification protocol (b)), or using preparative
HPLC (Purification protocol (a)), or by normal-phase
flash-chromatography (Purification protocol (c)). Reaction time for
Compounds B1 and B2 was 4 days; and for all other compounds, 2
days.
Procedure B:
Preparation of Compound B7 (3-[3-(2-Hydroxy-((S)-valyl
ester)-4,6-dimethyl-phenyl)-3-methyl-butyrylamino]-1-propanesulfonic
acid)
##STR00139##
[0347] Step 1: EDC (N-(3-dimethylaminopropyl)-W-ethylcarbodiimide)
(6.4 g, 33 mmol, 3 eq.) was added, at 0.degree. C., to a 150-mL dry
dichloromethane solution containing Boc-Val-OH (4.9 g, 22 mmol, 2
eq.), the silylated phenol (3.6 g, 11 mmol; ref., J. Med. Chem.,
2000, 43, 475-487), and DMAP (4-(dimethylamino)pyridine, 5.5 g, 45
mmol, 4 eq.). The reaction mixture was stirred at room temperature
overnight, then diluted with dichloromethane, and washed with a
saturated aqueous solution of NaHCO.sub.3, 1N HCl, and brine
subsequently. The organic layer was dried and concentrated to a
colorless oil residue. Purification of the residual material (flash
chromatography; using hexane/EtOAc, 95/5) gave the corresponding
intermediate (5.7 g, 99%) as a colorless oil.
[0348] Step 2: The intermediate from step 1 (5.7 g, 11 mmol) was
stirred in a mixture of THF-water-acetic acid (20 mL/20 mL/60 mL)
at room temperature for 3 h; then the solvent was removed and the
residue dried in vacuo. The residual material (the free alcohol)
obtained was used in the next step without further purification
[0349] Step 3: A solution of the alcohol (11 mmol, from step 2) in
dichloromethane (125 mL) was slowly added to a suspension of PCC
(pyridinium chlorochromate, 5.0 g, 23 mmol, 2.1 eq.) in dry
dichloromethane (125 mL). The reaction mixture was stirred at room
temperature overnight. Solvent was evaporated and the residue was
dissolved in a minimum amount of dichloromethane. The resulting
dichloromethane solution was passed through a silica gel column
using Hexane/EtOAc (50/50). Evaporation of the solvent gave the
corresponding aldehyde as yellow oil which was directly used in the
next step without further purification.
[0350] Step 4: A solution of 80% sodium chlorite (2.5 g, 28 mmol,
2.5 eq.) in water (10 mL) was added slowly to a solution of the
aldehyde (11 mmol, form step 3) and sodium dihydrogen phosphate
(818 mg, 6.8 mmol, 0.6 eq.) in acetonitrile (20 mL) and water (20
mL) at 0.degree. C. The mixture was stirred 1 h at 0.degree. C.
then at room temperature for 1 h. Sodium sulfite (1.5 g, 1 eq.) was
added to decompose peroxides, and the pH was adjusted to 2 with 1N
HCl solution. Reaction mixture was extracted twice with EtOAc. The
organic layers were washed with brine, dried, and concentrated.
Purification of the residual material (flash chromatography;
CH.sub.2Cl.sub.2/CH.sub.3OH, 100/0 to 95/5) gave the corresponding
carboxylic acid (3.4 g, 73%) as a foam.
[0351] Step 5: EDC (908 mg, 4.75 mmol, 2 eq.) was added to a
mixture of the carboxylic acid (1 g, 2 mmol; from step 4),
3-amino-1-propanesulfonic acid sodium salt (380 mg, 2.34 mmol) and
a catalytic amount of DMAP in DMF (10 mL). The reaction mixture was
stirred at room temperature overnight. Solvent was removed and the
residue was dried in vacuo to provide the corresponding derivative
of 3-amino-1-propanesulfonic acid which was used in the next step
without further purification.
[0352] Step 6: Trifluoroacetic acid (5 mL) was added to a solution
of the 3-amino-1-propanesulfonic acid derivative (2.4 mmol, from
step 5) in dichloromethane (5 mL) at room temperature. The reaction
mixture was stirred for 2 h, followed by evaporation of the
solvent. The resulted residue was purified (preparative HPLC;
acetonitrile/water, 5/95 to 70/30 in the presence of 0.01% TFA) to
yield, after lyophilization, the title compound (0.3 g, 29%) as a
white solid: .sup.1H NMR (500 MHz, D.sub.2O) .delta.1.04 (d, J=7
Hz, 3H), 1.07 (d, J=7 Hz, 3H), 1.39 (s, 3H), 1.45 (s, 3H),
1.55-1.58 (m, 2H), 2.11 (s, 3H), 2.43 (s, 3H), 2.45-2.58 (m, 5H),
2.98-3.02 (m, 2H), 4.26 (d, J=4 Hz, 1 H), 6.54 (d, J=1.5 Hz, 1 H),
6.93 (d, J=1.5 Hz, 1H).
Procedure C:
Preparation of Compound B14:
3-{[(3.alpha.,5.beta.,7.alpha.,12.alpha.)-3,7,12-trihydroxy-24-oxo-cholan-
-24-yl]amino}-1-propanesulfonic acid
##STR00140##
[0354] To a mixture of (+)-cholic acid (5.0 g, 12.2 mmol),
3-amino-1-propanesulfonic acid sodium salt (1.85 g, 11.5 mmol),
4-dimethylaminopyridine (72 mg, 0.6 mmol) in DMF (30 mL) was added
N-(3-dimethylaminopropyl)-N'-ethylcarbodimide hydrochloride (EDC,
4.68 g, 24.4 mmol). The reaction mixture was stirred at room
temperature overnight. The cloudy mixture was filtered through
sintered glass before the solvent was evaporated to dryness under
reduced pressure. The viscous residue was dissolved in water (30
mL). The solution was treated with Dowex Marathon C.TM. ion
exchange resin (strongly acidic, 30 g, pre-washed). The suspension
was stirred for 15 minutes before the resin was removed by
filtration. The filtrate was concentrated to dryness under reduced
pressure and dried in vacuo. The residue was triturated with
diethyl ether (1000 mL). The solid product was recovered by
filtration and dried in vacuo. The crude product was purified by
flash chromatography (Biotage.TM. SP1: 20-40% EtOH in
CH.sub.2Cl.sub.2) and the corresponding fractions were collected
and lyophilized, affording the title compound (178 mg, 3%); .sup.1H
NMR (D.sub.2O, 500 MHz) .delta. ppm 0.73 (s, 3H), 0.93 (s, 3H),
1.02 (m, 4H), 1.31 (m, 7H), 1.52 (d, 1H, J=14.5 Hz), 1.65 (m, 6H),
1.79 (m, 3H), 1.94 (m, 3H), 2.04 (m, 3H), 2.23 (m, 1 H), 2.31 (m, 1
H), 2.92 (m, 2H), 3.31 (m, 2H), 3.52 (m, 1H), 3.92 (s, 1H), 4.08
(s, 1H); .sup.13C NMR (D.sub.2O, 125 MHz) 8 ppm 12.31, 16.82,
22.33, 23.12, 24.30, 26.48, 27.47, 27.95, 29.37, 31.87, 32.71,
34.06, 34.54, 35.09, 35.33, 38.15, 38.49, 39.50, 41.29, 41.64,
46.27, 46.28, 48.73, 68.33, 71.69, 73.14, 177.44; m/z (ES.sup.+)
530; [.alpha.].sub.D=+25.7.degree. (c=0.005, water).
TABLE-US-00008 TABLE 7 Synthesis and characterization of exemplary
non-amino acid amide prodrugs according to the invention Synthetic
Purifying m/z (ES.sup.-) ID procedure protocol* (M - H, or M -
Na).sup..dagger. B1 A (a) 320.4 B2 A (a) 306.5 B3 A (b) 280.2 B4 A
(c) 280.3 B5 A (b) 238.0 B6 A (b) 525.0 B7 B (a) 441.3 B9 B (a)
491.4 B10 B (a) 457.3 B11 B** (a) 514.2 B13 B** (a) 548.1 *(a),
HPLC; (b), precipitation; (c), flash chromatography; (d),
filtration; (e), extraction; **Procedure B, replacing 3-APS by
N-glycyl-3-APS; .sup..dagger.the compounds were synthesized as acid
form, or as sodium salt form.
Example 1-D: Chemical Synthesis of Carbohydrate-Derived
Prodrugs
[0355] Accordingly, the following examples are presented to
illustrate how some carbohydrate-derived prodrugs according to the
invention compounds may be prepared.
Synthesis of Compound S1 Sodium Salt
##STR00141##
[0357] A suspension of glucose (2 g, 11.1 mmol) and the sodium salt
of 3APS (2.24 g, 11.1 mmol) in MeOH (10 mL) was refluxed for 30 min
before being cooled down to room temperature. After 24h of stirring
at room temperature, the solid was filtrated and washed twice with
MeOH (2.times.10 mL). The resulting solid was dried overnight under
high vacuum and afford the sodium salt of Compound S1 (3.1 g, 9.6
mmol, 86%) as a white solid. .sup.1H NMR (D.sub.2O) (500 MHz)
.delta. ppm 4.55 (d, J=4.4 Hz, 0.33H, .alpha.-anomer), 3.87 (d,
J=9.3 Hz, 0.66H, .alpha.--anomer), 3.74 (dd, J=12.2, 1.5 Hz,
0.66H), 3.70 (dd, J=12.7, 2.4 Hz, 0.33H), 3.61 (dd, J=12.2, 4.9 Hz,
0.33H), 3.56 (dd, J=12.2, 5.4 Hz, 0.66H), 3.53-3.49 (m, 1H), 3.33
(t, J=9.3 Hz, 0.66H), 3.25-3.20 (m, 1 H), 3.05 (t, J=8.8 Hz,
0.33H), 2.83 (m, 2.66H), 2.68 (m, 1 H), 2.57 (m, 0.33H), 1.78 (m,
2H). m/z (ES'') 300.0 (M-H).
Synthesis of Compound S2
##STR00142##
[0359] Methyl 6-bromo-6-deoxy-.alpha.-D-glucopyranoside was
Prepared According to Tetrahedron 1991, 28(47), 5185-5192
[0360] Step 1: A stirred suspension of bromide (1 g, 3.89 mmol) and
sodium azide (278 mg, 4.28 mmol) in DMF (10 ml) was stirred at
90.degree. C. for 5 days. After being cooled down to room
temperature, the solution was evaporated under vacuum and the
residue was purify by chromatography on silica gel (CHCl.sub.3/MeOH
95/5 to 70/30 linear gradient) to afford the desired azido (776 mg,
3.54 mmol , 91%) as a white solid.
[0361] Step 2: A solution of the previously prepared azido
derivative (776 mg, 3.54 mmol) in MeOH (10 ml) was degazed with
N.sub.2 for 10 min before a suspension of 10% Pd/C (50 mg) in
CHCl.sub.3 was added. After being stirred 2 h under H.sub.2
pressure (40 PSI), the solution was filtrated over a pad of
Celite.TM. (MeOH) and evaporated under vacuum and afforded the
desired amine (628 mg, 3.25 mmol, 92% crude) as a yellow oil. This
compound was used in the next step without further
purification.
[0362] Step 3: A solution of sultone (285 .mu.l, 3.25 mmol) in
CH.sub.3CN (5 ml) was added drop wise (over 30 min) to a refluxing
solution of the previously prepared amine (628 mg, 3.25 mmol) in a
2/1 mixture CH.sub.3CN/EtOH (10 ml). The resulting solution was
heated under reflux for 15 h before being cooled down to room
temperature and evaporated under vacuum. The residue was purified
by chromatography on silica gel (i-PrOH/H.sub.2O (0.5% NH.sub.4OH)
98/2 to 80/20 linear gradient). After Evaporation, the compound was
passed through a C-8 pad (H.sub.2O) and lyophilized and afforded
Compound S2 (450 mg, 1.43 mmol, 44% over two steps) as a white
solid. NMR .sup.1H (D.sub.2O) (500 MHz): 2.06(m, 2H), 2.92(t, J=7.0
Hz, 2H), 3.13 (m, 3H), 3.21 (t, J=9.5 Hz, 1H), 3.34 (s, 3H), 3.36
(dd, 12.5, 3 Hz, 1H), 3.48 (dd, J=9.5, 3.5 Hz, 1H), 3.56 (t, J=9.0
Hz, 1H), 3.77 (dt, J=9.0, 2.5 Hz, 1 H), 4.74 (d, J=3.5 Hz, 1 H). ES
(MS) 314.1 (M-H). [.alpha.].sub.D=+86.3 (c 1.0, H.sub.2O)
Procedure A: General Procedure for the Deprotection of 1,2,3,4- or
2,3,4,6-tetraacetate Glucose Derivative
[0363] To a stirred solution of the protected glucose derivative
was added enough of a solution of NaOMe (sodium methoxide, 0.5M in
MeOH) in order to obtain a basic pH (8-9, pH paper). The resulting
solution was stirred at room temperature until completion (the
reactions were generally followed by MS) before addition of twice
the initial volume of CH.sub.3CN. The resulting solid was then
filtrated and washed several time with CH.sub.3CN, acetone and
diethyl ether. The resulting solid was then passed trough a C8
column (0.5% NH.sub.4OH in H.sub.2O) and lyophilized to afford the
desired compound.
Synthesis of Compounds S3 and S4
##STR00143##
[0365] Step 1: A suspension of the sodium salt of
3-amino-1-propanesulfonic acid (398 mg, 2.47 mmol) and
glucopyranuronic anhydride (398 mg, 2.47 mmol) in DMF (15 mL) was
stirred 3 days at room temperature before evaporation of the
solvent under vacuum. The residue was purified by chromatography on
silica gel (CHCl.sub.3/MeOH 100/0 to 70/30 linear) to afford
compound S3 (719 mg, 1.49 mmol, 60%) as a white foam. .sup.1H NMR
(CD.sub.3OD, 500 MHz) .delta. ppm 1.96(m, 2H), 1.98(s, 3H), 2.01(s,
3H), 2.02(s, 3H), 2.09(s, 3H), 2.83(m, 2H), 3.31(m, 2H), 4.19(d,
J=9.5 Hz, 1H, H.sub.5), 5.12(t, J=8 Hz, 1H, H.sub.2), 5.19(t, J=10
Hz, 1H, H.sub.4), 5.38(t, J=9 Hz, 1H, H.sub.3), 5.87(d, J=8.5 Hz,
1H, H.sub.1). m/z (ES) 482.4 (M-H); =+6.2 (c 0.93, MeOH).
[0366] Step 2: Compound S3 (190 mg, 0.54 mmol) was treated
according to Procedure A to afford Compound S4 (150 mg, 0.48 mmol,
88%) as a white solid. .sup.1H NMR (D.sub.2O, 500 MHz) .delta. ppm
1.92 (m, 2H), 2.90 (m, 2H), 3.27 (t, J=8.5 Hz, 0.5H), 3.32 (m, 2H),
3.47-3.50 (m, 1.5H), 3.56 (dd, J=9.5, 4.0 Hz, 0.5H), 3.69 (t, J=9.0
Hz, 0.5H), 3.86 (d, J=7.0 Hz, 0.5H), 4.16 (d, J=10.0 Hz, 0.5H),
4..6-4.7 (0.5H, under water peak), 5.25 (d, J=3.5 Hz, 0.5H); m/z
(ES.sup.-) 314.4 (M-H).
Synthesis of Compound S5 Sodium Salt and Compound S6 Ammonium
Salt
##STR00144##
[0367] 2,3,4,6-Tetra-O-acetyl-D-glucose was prepared according to
J. Am. Chem. Soc. 1993, 115, 2260-2267.
[0368] Step 1: p-nitrophenolchloroformate (638 mg, 3.16 mmol) was
added to a stirred solution of tetraacetylglucose (1 g, 2.87 mmol)
and Et.sub.3N (800 .mu.l, 5.74 mmol) in CH.sub.2Cl.sub.2 (20 ml)
and the reaction was stirred overnight at room temperature. A 1N
aqueous solution of hydrochloric acid (10 ml) was added and the
layers were separated. The aqueous layer was extracted twice with
CH.sub.2Cl.sub.2 (20 ml) and the combined organic layer were washed
subsequently with a saturated solution of sodium carbonate (10 ml)
and a saturated solution of sodium chloride. The organic layer was
then dry over MgSO.sub.4, filtrated and the solvent was evaporated
under vacuum. The residue was purified by chromatography on silica
gel (Hex/EtOAc 90/10 to 5050, linear gradient) to afford the
desired carbonate (1.108 g, 2.16 mmol, 75%) as colorless solid.
[0369] Step 2: Pyridine (524 ml, 6.48 mmol) was added to a
suspension of the carbonate previously prepared (1.108 g, 2.16
mmol) and the sodium salt of 3APS (522 mg, 2.16 mmol). After 3 days
of stirring at room temperature, the solvent was evaporated under
vacuum and the residue was purified by chromatography on silica gel
(CHCl.sub.3/MeOH 100/0 to 80/20, linear gradient) to afford
Compound S5-Sodium salt (1.066 g, 2.07 mmol, 96%) as a white solid.
.sup.1H NMR (D.sub.2O, 500 MHz) 5 ppm 1.97 (s, 3H), 2.00 (s, 3H),
2.01 (s, 3H), 2.1 (m, 2H, hide), 2.05 (s, 3H), 2.83 (m, 2H), 3.25
(m, 2H), 3.98 (br d, J=8.0 Hz, 0.4H, H.sub.5b), 4.09 (t, J=10.5 Hz,
1H, H.sub.6), 4.17 (br d, J=10.2 Hz, 0.6H, H.sub.5a), 4.26-4.30 (m,
1H, H.sub.6), 4.99-5.12 (m, 2H, H.sub.2a, H.sub.2b, H.sub.4b,
H.sub.4a), 5.32 (t, J=9.5 Hz, 0.40H, H.sub.3b), 5.50 (t, J=9.9,
0.6H, H.sub.3a), 5.69 (d, J=8.4 Hz, 0.3H, H.sub.1b), 6.17 (d, J=3.5
Hz, 0.6H, H.sub.1a). m/z (MS) 512.5 (M-H).
[0370] Step 3: Compound S5 sodium salt (500 mg, 0.97 mmol) was
treated according to Procedure A to afford Compound S6-ammonium
salt (220 mg, 0.64 mmol, 66%) as a white solid. .sup.1H NMR
(D.sub.2O (500 MHz) .delta. ppm 1.80 (m, 2H), 2.80 (m, 2H), 3.15
(m, 2H), 3.30-3.37 (m, 1.5H), 3.41-3.43 (m, 1H), 3.68-3.53 (m, 3H),
3.75 (d, J=12.2 Hz, 0.5H), 5.26 (d, J=8.2 Hz, 0.5H, H.sub.1b), 5.82
(d, J=3.05 H2, 0.5H, H.sub.1a). m/z (ES) 344.4 (M-H).
Synthesis of the Sodium Salt of Compound S7
##STR00145##
[0371] 2-(p-nitrophenyl
carbamate)-ethyl-2,3,4,6-tetra-O-acetyl-,6-D-glucopyranoside was
prepared according to Org. Lett. 2000, 2(8), 1093-1096.
[0372] Step 1: 3APS-sodium salt (223 mg, 1.38 mmol) was added to a
stirring solution of p-nitrophenyl carbamate (643 mg, 1.16 mmol) in
DMF (7 mL). After 24 h of stirring at room temperature, the solvent
was evaporated under vacuum and the residue was purified by
chromatography on silica gel (CHCl.sub.3/MeOH 100/0 to 70/30,
linear gradient) and afforded the desired sulfonate (596 mg, 1.07
mmol, 92%) as a white solid.
[0373] Step 2: The 2,3,4,6-tetra-O-acetyl-D-glucose previously
prepared (596 mg, 1.07 mmol) was treated according to Procedure A
to afford Compound S7-sodium salt (260 mg, 0.67 mmol, 63%) as a
white solid. .sup.1H NMR (D.sub.2O, 500 MHz) .delta. ppm 1.91 (m,
2H, H11), 2.93 (t, J=7.5 Hz, 2H, H12), 2.24 (t, J=6.0 Hz, H10),
3.28 (t, J=9.0 Hz, 1H, H2), 3.34 (m, 2H, H8), 3.38 (t, J=9.5 Hz,
1H, H4), 3.45 (m1, 1H, H6a), 3.49 (dd, J=9, 9 Hz, 1H, H3), 3.7-3.77
(m, 2H, H6a, H7a), 3.91 (apparent d, J=11.5 Hz, H5, H7b), 4.46 (d,
J=8.0 Hz, H1). m/z (ES) 386.9 (M-H).
Synthesis of the Sodium Salt of Compounds S8 and S9
##STR00146##
[0374]
N-(9-Fluorenylmethoxycarbonyl)-3-O-(2,3,4,6-tetra-.beta.-acetyl-.be-
ta.-D-glucopyranosyl)-L-serine Pentafluorophenyl ester was prepared
according to J. Med. Chem. 1995, 38, 161-169.
[0375] Step 1: 3APS-sosium salt (258 mg, 1.60 mmol) was added to a
stirring solution of pentaflurophenyl ester (1200 mg, 1.45 mmol) in
DMF (15 mL). After 24 h of stirring at room temperature, the
solvent was evaporated under vacuum and the residue was purified by
chromatography on silica gel (CHCl.sub.3/MeOH 100/0 to 80/20,
linear gradient) to afford the desired sulfonate (1070 mg, 1.37
mmol, 94%) as a white solid.
[0376] Step 2: Piperidine (2.7 mL, 27 mmol) was added to a stirred
solution of previously prepared Fmoc serine derivative (1070 mg,
1.37 mmol) in DMF (15 mL). After stirred for 1 h, solvent was
evaporated under reduced pressure. The residue was purified by
chromatography on silica gel (CHCl.sub.3/MeOH 100/0 to 75/25,
linear gradient) to afford the desired amine Compound S8-sodium
salt (350 mg, 0.63 mmol, 46%) as a white solid.
[0377] Step 3: The 2,3,4,6-tetra-O-acetyl-D-glucose previously
prepared (350 mg, 0.63 mmol) was treated according to Procedure A
to afford Compound S9-sodium salt (210 mg, 0.54 mmol, 86%) as a
white solid. .sup.1H NMR (D.sub.2O, 500 MHz) 1.95 (m, 2H, H11),
2.94 (t, J=8.0 Hz, 2H, H12), 3.35 (dd, J=7.5, 9.0 Hz, 1H, H2),
3.36-3.41 (m, 3H, H4, H10), 3.42-3.50 (m, 2H, H3, H5), 3.73 (dd,
J=6.0, 1 H, 12.0 Hz, H6a), 3.92 (br d, J=12.0 Hz, 1 H, H6b), 3.96
(dd, J=4.5, 1H, 11.5 Hz, H8), 4.05 (t, J=4.5 Hz, 1H, H7a), 4.22
(dd, J=4.5, 11.5 Hz, 1H, H7), 4.47 (d, J=7.5 Hz, 1H, H1). m/z (ES)
387.25 (M-H).
Synthesis of the Sodium Salt of Compounds S14 and S15
##STR00147##
[0379] 1,2,3,4-tetra-O-acetyl-.alpha.-D-glucopyranoside was
prepared according to Org. Lett. 2006, 8, 2393-2396 and J. Am Chem.
Soc. 2000, 122, 12151-12157.
[0380] Step 1: p-Nitrophenolchloroformate (3 g, 14.8 mmol) was
added to a stirred solution of
1,2,3,4-tetra-O-acetyl-.alpha.-D-glucopyranoside (4.7 g, 13.4 mmol)
and triethylamine (3.7 ml, 26.8 mmol) in dichloromethane (100 mL).
The reaction mixture was stirred overnight at room temperature. A
1N aqueous solution of hydrochloric acid (30 mL) was added and the
layers were separated. The aqueous layer was extracted twice with
dichloromethane (100 mL) and the combined organic layers were
washed subsequently with a saturated solution of sodium carbonate
(50 mL) and then with a saturated solution of sodium chloride. The
organic layer was dried over magnesium sulfate, filtered and the
solvent was evaporated under vacuum. The residue was purified by
chromatography on silica gel (hexanes/ethyl acetate 90/10 to 50/50,
linear gradient), affording the corresponding carbonate (4.7 g,
68%) as a colorless solid.
[0381] Step2: The sodium salt of 3APS (2.22 g, 13.8 mmol) was added
to a solution of the carbonate previously prepared (4.7 g, 9.16
mmol) in N,N-dimethylformamide (50 mL). After 3 days of stirring at
room temperature, the solvent was evaporated under vacuum and the
residue was purified by chromatography on silica gel
(dichloromethane/methanol 100/0 to 70/30, linear gradient) and
afforded Compound S15-sodium salt (1.95 g, 41%) as a white solid
together with its 1-deactetylated derivative (1.21 g, 36%) as a
white solid: .sup.1H NMR (D.sub.2O, 500 MHz) .delta. ppm 1.91-2.02
(m, 11H), 2.07 (s, 2H), 2.17 (s, 1 H), 2.86 (m, 2H, H1), 3.24 (t,
J=8.0 Hz, 2H, H3), 3.99 (m, 0.7H, H6B), 4.10-4.20 (m, 2.3H, H5 and
H6a), 5.02 (m, 1H, H9), 5.08 (t, J=10.0 Hz, 0.7H, H7B), 5.13 (t,
J=9.5 Hz, 0.3H, H7a), 5.34 (t, J=9.5 Hz, 0.7H, H8.beta.), 5.44 (t,
J=9.5 Hz, 0.3H, H8.alpha.), 5.81 (d, J=8.0 Hz, 0.7H, H10.beta.),
6.28 (d, J=3.5 Hz, 0.3H, H10.alpha.); m/z (ES) 512.0 (M-H).
[0382] Step3: Compound S15-sodium salt (1.37 g, 2.67 mmol)) was
treated according to Procedure A to afford Compound S14-sodium salt
(520 mg, 1.51 mmol, 56%) as a white solid: .sup.1H NMR(D.sub.2O,
500 MHz) .delta. ppm 1.80 (m, 2H, H2); 2.81 (m, 2H, H1), 3.12 (m,
2.55H, H3 and H9.beta.); 3.31 (m, 1H, H7.alpha. and H7.beta.); 3.36
(m, 0.55H, H8.beta.); 3.41 (dd, J=10.0, 4.0 Hz, 0.45H, H9.alpha.);
3.48 (m, 0.55H, H6.beta.); 3.56 (t, J=9.0 Hz, 0.45H, H8.alpha.);
3.84 (brd, J=10.0 Hz, 0.45H, H6.alpha.), 4.10 (m, 1H, H5a), 4.23
(apparent t, J=12.5 Hz, 1H, H5b), 4.51 (d, J=8.0 Hz, 0.55H,
H10.beta.); 5.08 (d, J=4.0 Hz, 0.45H, H10.alpha.); m/z (ES) 344.0
(M-H).
Synthesis of the Sodium Salt of Compounds S16 and S17
##STR00148##
[0383] 2,3,4,6-Tetra-O-acetyl-D-glucose-1-propanol was prepared
according to J. Am. Chem. Soc. 1940, 62, 917-920.
[0384] Step 1: p-Nitrophenolchloroformate (2.3 g, 11.4 mmol) was
added to a stirred solution of 3-hydroxy-1-propyl
2,3,4,6-tetra-O-acetyl-.beta.-D-glucopyranoside (3.1 g, 7.64 mmol)
and triethylamine (2.12 mL, 11.44 mmol) in dichloromethane (60 mL)
and the reaction mixture was stirred overnight at room temperature.
Aqueous hydrochloric add (1N, 15 mL) was added and the layers were
separated. The aqueous layer was extracted 2 times with
dichloromethane (40 mL) and the combined organic layer were washed
subsequently with a saturated solution of sodium carbonate (15 mL)
and then with a saturated solution of sodium chloride. The organic
layer was then dried over magnesium sulfate, filtrated and the
solvent was evaporated under vacuum. The residue was purified by
chromatography on silica gel (hexanes/ethyl acetate 90/10 to 50/50,
linear gradient) to afford the corresponding carbonate (3.1 g, 71%)
as colorless solid.
[0385] Step2: The sodium salt of 3-APS (655 mg, 4.07 mmol) was
added to a solution of the carbonate previously prepared (1.55 g,
2.71 mmol) in N,N-dimethylformamide (50 mL). After 3 days of
stirring at room temperature, the solvent was evaporated under
vacuum and the residue was purified by chromatography on silica gel
(dichloromethane/methanol 95/5 to 70/30, linear gradient) to afford
a mixture of Compound S17 and p-nitrophenol (1.33 g) as a white
solid, which was used in next step without further
purification.
[0386] Step3: The crude Compound S17 (1.33 g) was treated according
to Procedure A to afford Compound S16-sodium salt (850 mg, 49% over
two steps) as a white solid: .sup.1H NMR (D.sub.2O, 500 MHz)
.delta. ppm 1.84-1.91 (m, 4H, H6+H2), 2.88 (m, 2H, H1), 3.18 (m,
2H, H3), 3.21 (t, J=8.5 Hz, 1H, H9), 3.33 (t, J=9.3 Hz, 1H, H11),
3.39 (m, 1H, H12), 3.44 (t, J=9.3 Hz, 1H, H10), 3.67 (dd, J=12.3,
5.8 Hz, 1H, H13a), 3.71 (m, 1H, H7a), 3.85 (dd, J=12.3, 2.0 Hz, 1H,
H13b), 3.94 (m, 1 H, H7b), 4.10 (m, 2H, H5), 4.39 (d, J=8.0 Hz, 1
H, H8); m/z (ES) 402.1 (M-H).
Example 1-E: Chemical Synthesis of Imine-Derived Prodrugs
[0387] Accordingly, the following examples are presented to
illustrate how some imine-derived prodrugs according to the
invention compounds may be prepared.
Synthesis of Compound M7 Sodium Salt
##STR00149##
[0389] Sodium 3-amino-1-propanesulfonate (0.64 g, 4.0 mmol) was
added to a solution of 4'-chloro-5-fluoro-2-hydroxy-benzophenone
(0.50 g, 2.0 mmol) in methanol (50 mL). The reaction mixture was
stirred under reflux for 4 h then concentrated under reduced
pressure. The residual material was purified by flash
chromatography (silica gel, chloroform: methanol 90:10 then 80:20)
to afford the title compound (0.51 g, 64%): .sup.1H NMR
(CDCl.sub.3, 500 MHz) .delta. 1.89 (m, 2H), 2.5 (t, J=7.0 Hz, 2H),
3.36 (t, J=7.0 Hz, 2H), 6.95 (m, 1 H), 6.95 (m, 1 H), 7.22 (m, 1H),
7.38 (d, J=8.0 Hz, 2H), 7.66 (d, J=8.0 Hz, 2H), 15.27 (s, 1H).
ES-MS (370 M-1).
Synthesis of Compound M7-sulfonamide
##STR00150##
[0391] Step 1: To a stirred solution of sodium azide (3.5 g, 50
mmol) in water (25 mL) was added a solution of 1,3-propane sultone
(6.1 g, 50 mmol) in acetone (25 mL). The reaction mixture was
stirred at room temperature for 24 h then concentrated to dryness.
The resulting solid was suspended in diethyl ether (100 mL) and
stirred at reflux for 1 h. The suspension was cooled to room
temperature and the solid was collected by filtration, washed with
acetone and diethyl ether, and dried under vacuum, affording of
3-azido-1-propanesulfonic acid (7.6 g, 80%).
[0392] Step 2: PCl.sub.5 (2.61 g, 12.53 mmol) was added to a
suspension of 3-azido-1-propanesulfonic acid (2.07 g, 12.53 mmol)
in toluene. The reaction mixture was stirred under reflux for 3 h.
After cooling to room temperature, the solvent was evaporated, and
the resulting material was used in the next step without further
purification.
[0393] Step 3: Ammonium hydroxide (28%) (10 mL) was added to a
solution of 3-azido-1-propanesulfonyl chloride (=2.29 g, 12.53
mmol; obtained in step 2) in ethanol (10 mL). The reaction mixture
was stirred at room temperature for 3 h then concentrated. The
residual material was passed through a short silica gel column
using hexanes:ethyl acetate as eluent to isolate
3-azido-1-propanesulfonamide (1.5 g, 86%).
[0394] Step 4: 3-Azido-1-propanesulfonamide (1.5 g, 10.86 mmol;
obtained from step 3) was dissolved in water/ethanol (10 mL/10 mL),
followed by addition of 10% Pd/C (0.2 g). The resulting suspension
was stirred under atmospheric pressure of H.sub.2 for 5 h. The
insoluble material was removed by filtration; and the filtrate was
concentrated. The residual material was suspended in hydrogen. The
suspension was filtered and the resulting solid was washed with
ethanol and diethyl ether, dried under high vacuum, affording
3-amino-1-propanesulfonamide (1.2g, 80%).
[0395] Step 5: 3-Amino-1-propanesulfonamide (0.55 g, 4 mmol; from
step 4) was added to a solution of
4'-chloro-5-fluoro-2-hydroxy-benzophenone (1 g, 4 mmol) in methanol
(50 mL). The reaction mixture was stirred under reflux for 5 h then
concentrated under reduced pressure. The residual material was
purified by column chromatography (silica gel,
dichloromethane:methanol 90:10 then 80:20). The corresponding solid
(after removal of solvent) was recrystallized in diethyl ether to
afford
3-{[(1E)-(4-chlorophenyl)(5-fluoro-2-hydroxyphenyl)methylene]amino}propan-
e-1-sulfonamide (0.75 g, 51%). .sup.1H NMR (CDCl.sub.3, 500 MHz)
.delta. 2.21 (m, 2H), 3.24 (t, J=7.0 Hz, 2H), 3.47 (t, J=7.0 H,
2H), 4.63 (bs, 2H), 6.93 (m, 1H), 6.95 (m, 1H), 7.04 (m, 1H), 7.13
(d, J=8.2 Hz, 2H), 7.54 (d, J=8.2 Hz, 2H), 14.71 (s, 1H). ES-MS
(369 M-1).
Example 2: In Vitro Stability and Metabolism
[0396] In vitro stability of exemplary prodrugs of the invention
was tested in water, in an acidic aqueous solution (pH: 1.5), in
PBS, in human and mouse microsomes, and in human and mouse whole
blood.
A. Stability in Water, at pH: 1.5 and PBS
[0397] Stability of exemplary compounds was determined in water,
aqueous acidic solution (pH 1.5, HCl) and PBS (phosphate buffered
saline) solution using ESI-MS (electrospray ionization mass
spectrometry) as the detecting instruments. In general a 2 .mu.g/mL
pro-drug solution containing 1 .mu.g/ml IS (internal standard) was
prepared and incubated for 60 min. For water stability the
incubation was performed at room temperature and for stability in
acidic solution and in buffer. The incubation temperature was
37.degree. C. Samples were analyzed for prodrug content at time
points 0 and 60 min. using MS. The % changes in peak area ratio
after 60 minutes for each test compound tested are calculated using
the average values from six replicate runs. The compounds tested
included Compounds A1 to A19, Compounds B5 and B6 and Compounds C1
to C26. Except for C26 which was found unstable at pH 1.5 and in
PBS, all other compounds were judged to be stable under all
conditions tested with less than about 15%-20% concentration change
after 60 minutes.
B. Metabolism in Mouse and Human Microsomes
[0398] Microsomal stability of Compounds A1, A2, A3, C17, C18 and
C19 was determined in duplicate, in presence of pooled mouse or
human liver microsomes for up to 60 minutes at 37.degree. C.
Briefly, microsomes were diluted to achieve a concentration of 1.0
mg/mL in PBS buffer (pH 7.4) containing 3 mM MgCl.sub.2 and 1 mM
EDTA. Compounds (10 .mu.M) and microsomes were pre-incubated for a
period of 5 minutes before the enzymatic reaction was started by
addition of co-factors (1 mM NADPH- and 2 mM UDPGA in PBS buffer).
After a 1-hour incubation period, the reaction was stopped by the
addition of ice cold acetonitrile. For time 0 samples, the reaction
was stopped with acetonitrile before the addition of the
co-factors. Analysis of extracted samples was achieved using HPLC
with MS detection. Several types of HPLC columns and mobile phases
were used depending of the polarity of the compound. The compound
stability was determined by the % of compound remaining at 60
minutes (peak response of compound at 60 minutes/peak response at 0
minutes.times.100). Four of the compounds tested (three amino acid
prodrugs A1, A2, A3, and the carbamate prodrug C19) were found
stable, with over 90% of the compounds remaining after 60 minutes
in presence of mouse or human microsomes (data not shown). Compound
C17 was found less stable with between 20 and 35% of the prodrug
remaining after 60 minutes in presence of mouse or human
microsomes, while carbamate C18 showed moderate stability with
between 75 and 80% of the prodrug remaining under the same
conditions.
C. Mouse and Human Whole Blood Stability
[0399] Test compounds were incubated for a total of 240 minutes at
37.degree. C. in whole mouse and whole human blood. The compounds
were added at time-point 0 and sample aliquots were withdrawn at
each time point (usually 0, 60 and 240 minutes). The samples were
extracted using protein precipitation. Analysis of extracted
samples was achieved using HPLC with MS detection. Several types of
HPLC columns and mobile phase were used depending of the polarity
of the compound. The compound stability was determined by the % of
compound remaining at 240 minutes (peak response of compound at 240
minutes/peak response at 0 minutes.times.100). Results are
summarized in Table 8.
TABLE-US-00009 TABLE 8 Stability in mouse and human whole blood
Blood stability (% of compound remaining after 240 min.) ID Human
Blood Mouse Blood A1 ND + A2 ND ++ A3 ND ++ A4 + +++ A5 + + A6 ++ +
A7 +++ +++ A8 + ++ A9 +++ +++ A10 ++ ++ A11 +++ +++ A12 +++ +++ A13
+++ +++ A14 +++ +++ A15 ++ + A16 +++ ++ A18 + + A19 + + B3 +++ ++
B4 +++ +++ B5 +++ +++ B6 +++ +++ C1 + + C4 +++ ++ C5 + + C7 +++ +
C8 +++ +++ C9 ++ ++ C10 + + C11 +++ +++ C12 ++ + C13 + + C14 +++ ++
C15 ++ + C16 ++ + C17 ND + C18 ND + C19 ND + C20 ++ + C21 ++ + C22
++ + C23 ++ + C24 ++ + +: <30%, ++: 30-75%, +++: >75%; ND:
not determined
[0400] These data illustrate the use of these compounds as
prodrugs, as they are converted to 3APS in the blood.
Example 3: Pharmacokinetics in Mice
A. Bioavailability of Exemplary Compounds
[0401] Selected exemplary compounds were tested for bioavailability
in mice. Bioavailability estimates are performed for 3APS after
administration of molar equivalent the selected compounds. At a
specific time point following drug administration, one blood sample
(approximately 1 ml) is collected from each of 3 animals from the
inferior vena cava. The animals are anesthetized with isoflurane
before blood collection (approximately 45 sec).
[0402] Samples are collected at 5, 30, 60, 120, 180, 240 and 360
min post intravenous administration and at 15, 30, 60, 120, 180,
240 and 360 min post oral administration. One animal is used to
obtain a baseline sample (pre-dose sample). Blood samples are
collected into Sarstedt.TM. micro tubes (EDTA KE/1.3 ml), kept on
ice until centrifugation at 4.degree. C. at a minimum speed of 3000
rpm (1620G) for 10 min. Plasma samples are transferred into
Eppendorf.TM. tubes, immediately placed on dry ice and stored at
-80.degree. C. Plasma samples are stored frozen at -20.degree. C.
pending analysis.
[0403] Compounds in mouse plasma are extracted using protein
precipitation. Quantitation of 3APS in mouse plasma matrix is
achieved using LC-MS detection. Sample concentration is calculated
using a calibration curve. Bioavailability results are summarized
in Table 9.
TABLE-US-00010 TABLE 9 Bioavailability of selected compounds in
mice Bioavailability (F) in mice * ID (+: <25%, ++: 25-35%, +++:
>35%) A (3APS) ++ A1 ++ A2 +++ A3 + A4 +++ A6 ++ A7 +++ A13 +++
A18 +++ C9 + C13 + C14 + C15 + C16 + C17 + C18 + C19 ++ C21 + C22 +
C25 + * Calculated from the concentration of 3APS, 6 hours after
administration of the tested compound. The calculated F value
represents the Ratio (in percentage) of the AUC p.o. of the
compound tested over the AUC i.v. of 3APS, based on the observation
of 3APS.
[0404] As shown in Table 9, all the compounds tested were capable
of delivering measurable quantities of 3APS. Compounds A2, A4, A7
and A18 were helpful in increasing the bioavailability of 3APS
suggesting that they were more readily absorbed than 3APS or were
able to prevent first-pass metabolism of 3APS. Although not shown,
Compounds A3, C13, C14, C16, C17, C21, C22 and C25 had a measured
T.sub.max 4 times to 16 times longer that 3APS (0.25h), suggesting
a significant improvement in the pk profiles of 3APS using those
compounds.
B. PK Brain and Plasma Levels of Oral Compound A2 and 3-APS
[0405] Compounds A2 and 3-aminopropanesulfonic acid were tested for
pharmacokinetic parameters in mice. Parmacokinetic parameters
(Cmax, Tmax, T1/2, AUC) are evaluated for 3APS after administration
of a molar equivalent of each compound. Blood samples
(approximately 1 ml) and brain samples are collected from each of 3
animals at time points 5, 15, 30 minutes, 1, 2, 4, 6, 12, and 24
hours. The results analyzed from plasma samples and brain
homogenates are summarized in Table 10. Relative bioavailability (F
%) of Compound A2 and 3-APS were respectively of 51% and to 32%. A
2-fold increase in plasma concentration (Cmax) of 3-APS was
observed when orally administering Compound A2 compared to 3-APS.
Brain concentration of 3-APS was observed after oral administration
of 0.18 mmol/kg for Compound A2, whereas the concentration could
not be quantified after oral administration of the same molar
equivalent of 3-APS.
TABLE-US-00011 TABLE 10 PK data on 3-APS analysis following oral
administration of 25 mg/kg (0.18 mmol/kg) and 250 mg/kg (1.80
mmol/kg) equivalent of 3-APS Plasma Brain Dose Cmax Tmax T1/2 Cmax
Tmax T1/2 ID (mmol/kg) AUC (ng/mL) (h) (h) AUC (ng/mL) (h) (h)
3-APS 0.18 6427 1768 0.5 4.9 BLLQ BLLQ N/A N/A A2 0.18 10135 3435
0.5 2.8 557 148 2.0 3.9 A2 1.80 140661 35451 0.5 2.8 9772 1068 2.0
12.4 BLLQ: below the lower limit of quantification N/A: not
applicable
Example 4: Pharmacokinetic Analysis of 3APS and Associated
Metabolism
Example 4A: Metabolic Profiling of .sup.14C-3APS in Mice, Rats and
Dogs
[0406] Three single dose studies were conducted in mice, rats and
dogs to determine the metabolic profile of .sup.14C-3APS in plasma,
urine and feces. In the first study, twenty-seven male CD-1 mice
received a single dose of 100 mg/kg (20 .mu.Ci/animal) of
.sup.14C-3APS by oral gavage. Blood samples (3 animals/time point)
were collected for 12 hr following drug administration while urine
and feces (3 animals/time point) samples were collected for 96 hr.
In the second study, eight male Sprague-Dawley rats received a
single dose of 100 mg/kg (50 .mu.Ci/animal) of .sup.14C-3APS by
oral gavage while in the third study, three male Beagle dogs
received a single dose of 100 mg/kg (30 .mu.Ci/kg) of .sup.14C-3APS
by oral gavage. For the rat and dog studies, blood samples were
collected for 24 hr following drug administration while urine and
feces samples were collected for 72 hr. All samples were analyzed
for total radioactivity using appropriate sample preparation
procedures and scintillation counting. Plasma and urine samples
were also analyzed for SAPS and 3APS metabolites
(2-carboxyethanesulfonic acid, 3-hydroxy-1-propanesulfonic acid and
3-acetylamino-1-propanesulfonic acid) concentrations using
qualified HPLC and MS/MS methods.
[0407] Following oral administration of 100 mg/kg .sup.14C-3APS to
mice and rats, mean maximum plasma concentrations of total
radioactivity and 3APS were reached at approximately 30 minutes
post-dose (Table 11). Thereafter, plasma concentrations of total
radioactivity and 3APS declined in a multi-phasic manner with
apparent terminal half-lives of approximately 2 and 6 h for mice
and rats, respectively. Mean maximum plasma concentration of
2-carboxyethanesulfonic acid was achieved at 120 to 240 h
post-dose. Thereafter, plasma concentrations declined in a
multi-phasic manner with an apparent terminal half-life of
approximately 2 h and 4 h for mice and rats, respectively.
[0408] Following oral administration of 100 mg/kg .sup.14C-3APS to
dogs, maximum plasma concentration of total radioactivity and 3APS
were reached at approximately 30 minutes post-dose, whereas maximum
plasma concentration of 2-carboxyethanesulfonic acid was achieved
at 720 minutes post-dose (Table 11). Thereafter, plasma
concentrations of total radioactivity and 3APS declined in a
multi-phasic manner. The mean apparent terminal half-lives were
approximately 35 h and 5 h for total radioactivity and 3APS,
respectively.
[0409] For all species, the majority of total radioactivity was
associated with 3APS and 2-carboxyethanesulfonic acid (Table 12).
Based on AUC.sub.0-.infin. values, 3APS accounted for approximately
60% of total radioactivity while 2-carboxyethanesulfonic acid
accounted for 30% in mice and rats. In dogs, 3APS accounted for
approximately 54% of total radioactivity while
2-carboxyethanesulfonic acid accounted for approximately 67%. 3APS
and 2-carboxyethanesulfonic acid AUC.sub.0-.infin. constituted
approximately 90% (mouse and rat) and approximately 121% (dog) of
the total radioactivity indicating that 2-carboxyethanesulfonic
acid is the major metabolite of 3APS in the mouse, rat and dog.
[0410] For all species, total radioactivity was quantitatively
recovered in urine and feces with approximately 75 to 90% of the
administered dose recovered in 72 h (rat and dog) or 96 h (mouse).
The major route of excretion of total radioactivity was via
urine.
[0411] On average, 60% of the dose was excreted in urine as total
radioactivity in all species. Based on the total amount of
radioactivity excreted in urine, approximately 30% was excreted as
3APS while 2-carboxyethanesulfonic acid accounted for another 63%
to 77% in mouse and dog. In rats, 3APS and 2-carboxyethanesulfonic
acid accounted for 59% and 62% of total radioactivity,
respectively. On average the two metabolites
3-hydroxy-1-propanesulfonic acid and
3-acetylamino-1-propanesulfonic acid represented less than 3% of
the total radioactivity in all species (Table 11). The urinary
cumulative amount of 3APS and 2-carlDoxyethanesulfonic acid
accounted for approximately 90 to 110% of that determined for total
radioactivity, once again suggesting that 2-carboxyethanesulfonic
acid is the major metabolite of 3APS in the mouse, rat and dog.
TABLE-US-00012 TABLE 11 Pharmacokinetic Parameters of Total
Radioactivity, 3APS and 2-carboxyethanesulfonic acid Following
Single Oral Administration of 100 mg/kg .sup.14C-3APS in Mice, Rats
and Dogs Parameter Mouse.sup.1 Rat Dog Total Radioactivity
C.sub.max (.mu.mol eq/mL) 0.126 0.228 0.249 T.sub.max (min) 30 30
31 AUC.sub.0-.tau. (.mu.mol eq min/mL) 24.4 43.3 45.4
AUC.sub..infin. (.mu.mol eq min/mL) 25.0 45.2 108 T.sub.1/2 (h)
2.14 6.02 35.7 3APS C.sub.max (.mu.mol/mL) 0.0977 0.218 0.250
T.sub.max (min) 30 30 31 AUC.sub.0-.tau. (.mu.mol min/mL) 15.5 26.7
24.5 AUC.sub..infin. (.mu.mol min/mL) 15.7 27.6 25.3 T.sub.1/2 (h)
1.72 6.43 5.04 2-carboxyethanesulfonic acid C.sub.max (.mu.mol/mL)
0.018 0.0234 0.0312 T.sub.max (min) 120 240 720 AUC.sub.0-.tau.
(.mu.mol min/mL) 7.26 12.7 30.5 AUC.sub..infin. (.mu.mol min/mL)
7.56 13.6 NC T.sub.1/2 (h) 2.33 3.99 NC .sup.1PK parameters were
derived using the mean plasma concentration-time profiles NC: Not
calculated
TABLE-US-00013 TABLE 12 Percentage of 3APS, 2-carboxyethanesulfonic
acid, 3-acetylamino- 1-propanesulfonic acid and
3-hydroxy-1-propanesulfonic acid in Plasma and Urine Following
Single Oral Administration of 100 mg/kg .sup.14C-3APS in Mice, Rats
and Dogs % of Total Radioactivity 2-carboxy- 3-acetylamino-
3-hydroxy- ethane- 1-propane- 1-propane- sulfonic sulfonic sulfonic
3APS acid acid acid Mouse Plasma* 63 30 -- -- Urine.sup..dagger. 30
62 3.1 0.4 Rat Plasma* 61 30 -- -- Urine.sup..dagger. 59 62 2.3 0.3
Dog Plasma* 54 67 -- -- Urine.sup..dagger. 29 77 0.01 0.3
*Calculated as [AUC0-.infin. 3APS or metabolites/AUC total
radioactivity)] (or using AUC0-t if AUC0-.infin. could not be
reliably estimated) .sup..dagger.Calculated as [Amount Excreted
3APS or metabolites/AUC total radioactivity)]
Example 4B: Absorption, Excretion and Plasma Kinetics of
.sup.14C-3APS in Humans
[0412] Following the identification of 3APS metabolites, plasma and
urine samples from this human AME study were reanalyzed for 3APS
and 3APS metabolite (2-carboxyethanesulfonic acid,
3-hydroxy-1-propanesulfonic acid and
3-acetylamino-1-propanesulfonic acid) concentrations using
qualified HPLC and MS/MS methods to determine the metabolic profile
of .sup.14C-3APS in human.
[0413] Following oral administration of .sup.14C-3APS to healthy
subjects, maximum plasma concentration of total radioactivity and
3APS were reached at approximately 1 to 1.25 hours post-dose,
whereas maximum plasma concentration of 2-carboxyethanesulfonic
acid was achieved at 6.5 hours. In plasma, the majority of total
radioactivity was associated with 3APS and 2-carboxyethanesulfonic
acid. Based on AUC.sub.0-.tau. values, 3APS accounted for
approximately 48% of total radioactivity while
2-carboxyethanesulfonic acid accounted for 49%. 3APS and
2-carboxyethanesulfonic acid AUC.sub.0-.tau. constituted
approximately 97% of the total radioactivity indicating that
2-carboxyethanesulfonic acid is the major metabolite of 3APS in
human plasma.
[0414] Based on the total amount of radioactivity excreted in
urine, approximately 15% was excreted as 3APS while
2-carboxyethanesulfonic acid accounted for another 79%. The urinary
cumulative amount of 3APS and 2-carboxyethanesulfonic acid
accounted for approximately 94% of that determined for total
radioactivity, once again suggesting that 2-carboxyethanesulfonic
acid is the major metabolite of 3APS.
Example 4C: Comparative Pharmacokinetic Parameters of SAPS and
2-carboxyethanesulfonic Acid Following a Single Oral and IV
Administration of .sup.14C-3APS to Rats
[0415] The purpose of this study was to investigate the absorption,
metabolism and excretion profiles of .sup.14C-3APS following a
single intravenous bolus and oral administration to rats.
Thirty-six male Sprague-Dawley rats received a single 100 mg/kg
(.about.50 .mu.Ci/animal) dose of .sup.14C-3APS by an IV bolus
injection (water or isotonic saline solution) and an additional 36
male rats received the same dose level by oral gavage (in water).
Blood, urine, feces, brain and CSF samples were collected for up to
72 hr following dose administration. Plasma, urine, brain and CSF
concentrations of 3APS and 2-carboxyethanesulfonic acid (3APS major
metabolite) were measured using LC and MS/MS detection method.
Plasma, urine, feces, brain and CSF samples were analyzed for total
radioactivity using appropriate sample preparation procedures and
scintillation counting.
[0416] Based on AUC.sub.0-.infin. values, after IV administration,
3APS accounted for 89% of total radioactivity and
2-carboxyethanesulfonic acid only about 9%. On the other hand,
after oral administration, 3APS accounted for about 68% of total
radioactivity and 2-carboxyethanesulfonic acid about 26%. Using
those data, it is possible to calculate a metabolite-to-parent
ratio of the exposure of about 0.1 following IV administration and
a ratio of 0.38 following oral administration. This higher
metabolite-to-parent ratio of the exposure following oral
administration when compared to IV is consistent with an intestinal
first-pass metabolism.
TABLE-US-00014 TABLE 13 Comparison of Systemic Exposure of 3APS and
2-carboxyethanesulfonic acid versus Total Radioactivity following a
Single IV and Oral Administration of 14C-3APS in Rats
AUC.sub.0-.infin.(nmol h/mL).sup.# % (3APS and 2-carboxy- %
(2-carboxy- 2-carboxy- ethane- Total ethane- ethane- sulfonic
Radio- sulfonic sulfonic Animal 3APS acid activity acid)* acid)**
IV 1001 1528 105 1625 6.5 100.5 1002 1420 144 1588 9.1 98.5 1003
1591 184 1883 9.8 94.3 1004 1147 125 1266 9.9 100.5 Mean 1422 140
1591 8.8 98.4 .+-.SD 196.2 33.7 253.0 1.60 2.93 % CV 13.8 24.1 15.9
18.1 2.98 PO 3001 610 232 874 26.5 96.3 3002 539 153 714 21.4 96.9
3003 407 177 628 28.2 93.0 3004 471 229 781 29.3 89.6 Mean 507 198
749 26.4 94.0 .+-.SD 87.4 39.1 104 3.49 3.37 % CV 17.3 19.8 13.9
13.2 3.59 .sup.#AUC.sub.0-.infin. expressed as nmol eq h/mL for
total radioactivity *Calculated as [(AUC.sub.0-.infin.
2-carboxyethanesulfonic acid/AUC total radioactivity)*100]
**Calculated as [(AUC.sub.0-.infin. 3APS + AUC.sub.0-.infin.
2-carboxyethanesulfonic acid)/AUC total radioactivity] *100
Example 4D: Comparative Pharmacokinetics Parameters of 3APS and
2-carboxyethanesulfonic acid Following a Single Oral, Intravenous
and Portal Administration of 3APS in Rats
[0417] The purpose of this study was to compare the pharmacokinetic
profile of 3APS following a single dose administration either
orally, intravenously or into the portal vein to male
Sprague-Dawley rats. The oral, intravenous and portal routes of
administration were selected to determine the intestinal and
hepatic first-pass effects in the rat. Three groups of 4 male
Sprague-Dawley rats were assigned to receive a single dose of 250
mg/kg 3APS by different routes of administration. One group
received 3APS as an IV bolus administration (in water or isotonic
saline solution), one group by oral gavage (in water) and the last
group via a catheter into the portal vein (in water or isotonic
saline solution). Blood samples were collected for 24 hours
following dose administration. Plasma concentrations of 3APS and
2-carboxyethanesulfonic acid (the major metabolite of 3APS) were
determined using LC and MS/MS method.
[0418] Following oral administration, maximum plasma concentrations
(C.sub.max) were generally reached within 1 hour for 3APS and its
bioavailability based on the AUC.sub..infin. was calculated to be
about 38%.
[0419] The results obtained confirmed that there is an important
metabolism of 3APS. More particularly, based on a comparison
between the systemic exposures following hepatoportal and
intravenous administrations, metabolism of 3APS associated with
hepatic first-pass was estimated to be 24%. By comparison between
the systemic exposures following oral and hepatoportal
administrations, metabolism of 3APS associated with intestinal
first-pass was estimated to be 43%. This study also showed that the
oral administration of 3APS generated 50% more metabolite than the
intravenous administration which is consistent with an intestinal
first-pass metabolism.
Example 5: In Vitro Metabolism of 3APS in Primary Rat Neuron
Culture and Organotypic Hippocampal Slice Culture
[0420] The metabolism of 3APS was also studied in vitro in
different types of cellular models. In some cases, the metabolism
of 3APS was compared with that of .gamma.-amino butyric acid
(GABA).
[0421] The results obtained demonstrated that incubation of 3APS
(400 .mu.M) in primary rat neuron culture media produced
2-carboxyethanesulfonic acid as a metabolite. The conversion of
3APS to 2-carboxyethanesulfonic acid was time-dependent and cell
concentration-dependent. Incubation of 3APS (400 .mu.M initial
concentration) for six days in the cell culture media (containing
800,000 cells) produced with 48 .mu.M of 2-carboxyethanesulfonic
acid. Under the same experimental conditions, 5.4 .mu.M succinic
acid was detected starting from GABA (400 .mu.M initial
concentration).
[0422] The conversion of 3APS to 2-carboxyethanesulfonic acid in
the primary neuron culture media was significantly inhibited by
vigabatrin, the latter a classic GABA transaminase inhibitor.
Nialamide, a monoamine oxidase inhibitor, also reduced the
formation of 2-carboxyethanesulfonic acid (from 3APS) but to a
lesser extent. In contrast, gabapentin (known to increase GABA
concentration in the brain) had no significant effect on the
conversion of 3APS to 2-carboxyethanesulfonic acid.
[0423] In another in vitro model employing organotypic hippocampal
slice culture, the conversion of 3APS to 2-carboxyethanesulfonic
acid was time-dependent. More than 60% of 3APS was converted to
2-carboxyethanesulfonic acid after 3-day incubation in the culture
media. 2-carboxyethanesulfonic acid was also detected after
incubation of 3APS in human hepatocyte (HepG2) culture media.
[0424] It is understood that the examples and embodiments described
herein are for illustrative purposes only and that various
modifications or changes in light thereof will be suggested to
persons skilled in the art and are to be included within the spirit
and purview of this application and scope of the appended
claims.
* * * * *