U.S. patent application number 11/147150 was filed with the patent office on 2006-06-01 for methods and compositions to treat glycosaminoglycan-associated molecular interactions.
Invention is credited to Francine Gervais, Allan M. Green, Robert Kisilevsky.
Application Number | 20060116347 11/147150 |
Document ID | / |
Family ID | 26788901 |
Filed Date | 2006-06-01 |
United States Patent
Application |
20060116347 |
Kind Code |
A1 |
Kisilevsky; Robert ; et
al. |
June 1, 2006 |
Methods and compositions to treat glycosaminoglycan-associated
molecular interactions
Abstract
Therapeutic compounds and methods for inhibiting a
glycosaminoglycan (GAG)-associated molecular interaction in a
subject, whatever its clinical setting, are described.
Inventors: |
Kisilevsky; Robert;
(Kingston, CA) ; Green; Allan M.; (Cambridge,
MA) ; Gervais; Francine; (Ile Bizard, CA) |
Correspondence
Address: |
LAHIVE & COCKFIELD
28 STATE STREET
BOSTON
MA
02109
US
|
Family ID: |
26788901 |
Appl. No.: |
11/147150 |
Filed: |
June 6, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10690020 |
Oct 20, 2003 |
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11147150 |
Jun 6, 2005 |
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09970148 |
Oct 2, 2001 |
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10690020 |
Oct 20, 2003 |
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09362505 |
Jul 27, 1999 |
6310073 |
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09970148 |
Oct 2, 2001 |
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60094454 |
Jul 28, 1998 |
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Current U.S.
Class: |
514/54 |
Current CPC
Class: |
A61K 31/472 20130101;
A61K 31/185 20130101; A61K 31/737 20130101; A61K 31/437 20130101;
Y02A 50/478 20180101 |
Class at
Publication: |
514/054 |
International
Class: |
A61K 31/737 20060101
A61K031/737 |
Claims
1. A method of treating a condition associated with a
glycosaminoglycan-associated molecular interaction in a subject,
comprising administering to a subject a therapeutically effective
amount of a therapeutic compound for modulating said
glycosaminoglycan-associated molecular interaction, said
therapeutic compound having the formula: QY.sup.-X.sup.+].sub.n
wherein Y.sup.- is an anionic group at physiological pH; Q is a
carrier molecule; X.sup.+ is a cationic group; and n is an integer
selected such that the biodistribution of said therapeutic compound
for an intended target site is not prevented while maintaining
activity of the therapeutic compound, or a pharmaceutically
acceptable salt or ester thereof, such that said
glycosaminoglycan-associated molecular interaction is modulated and
said condition is treated.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. Pat. No.
09/970,148, filed Oct. 2, 2001, which is a continuation of U.S.
Pat. No. 09/362,505, filed Jul. 27, 1999, which claims the benefit
of priority under 35 U.S.C. 119(e) to copending U.S. Provisional
Application 60/094,454, filed on Jul. 28, 1998, the entire contents
of which is incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] Glycosaminoglycans (GAGs) have been shown to be involved
with the early steps of the infectious process associated with
several pathogens. For example, it is believed that sulfated
proteoglycans are used by the infectious agents as anchors or
adsorption moieties for invasion of the host cells. Several
bacterial and viral infectious agents have been found to use
extracellular membrane components, such as GAGs, to access host
cells.
[0003] Heparan sulfate and/or other sulfated GAGs have been
suggested to be involved in the infection process by certain
bacteria such as Streptococcus pyogenes associated with acute
rheumatic fever and poststreptococcal glomerulonephritis, Chlamydia
trachoinatis, Staphylococcus aureus and Pseudomonas aeruginosa
(cystic fibrosis), Legionella pneumophila (Legionnaire's disease),
Bordetella pertussis (whooping cough), and Mycoplasma pneumoniae.
As one example, Streptococcus pyogenes surfaces bind fibronectin,
laminin, fibrinogen, nonspecific immunoglobulins A and G,
.alpha.2-macroglobulin, .beta.2-microglobulin and albumin.
Bacterial components do not bind to epithelial or endothelial cells
of the kidney but accumulate on the proteoglycan-rich regions that
connect these cells to the underlying connective tissue. Another
example, Chlamydia trachomatis, is one of the most common sexually
transmitted bacterial pathogens in the world. Infection appears to
be facilitated by binding of a heparan sulfate-like GAG present on
the surface of chlamydia, to a heparan sulfate receptor on the
target cell.
[0004] Certain types of viri, Herpesviridae, are believed to be
associated with HSPG during the infectious process. These viri
appear to interact with a cells surface through GAGs found on the
proteoglycans of the cell plasma membrane. These GAGs are similar
to heparin. Cytomegalovirus (CMV) and Herpes simplex (HSV-1 and
HSV-2) are two of the viri which are believed to infect cells via
cell surface GAGs.
[0005] Although certain agents have been used to suppress infection
of hosts by pathogens, there are limitations to their use. For
example, the widespread use of antibiotics has increasingly led to
the problem of resistant pathogens whose growth can no longer be
inhibited by known antibiotics. Thus, the appearance of multi-drug
resistant pathogens has prompted a search for new classes of
compounds which are structurally and/or functionally different from
existing drugs. Drugs having new mechanisms of action could be
effective against resistant pathogens, where conventional drugs can
no longer be used.
SUMMARY OF THE INVENTION
[0006] Methods and compositions which are useful in the treatment
of conditions related to glycosaminoglycan (GAG)-associated
molecular interactions are presented herein.
[0007] In one aspect the invention relates to methods for treating
a condition related to a glycosaminoglycan-associated molecular
interaction in a subject. The method includes administering to the
subject a therapeutically effective amount of a therapeutic
compound having the formula: QY.sup.-X.sup.+].sub.n (I)
[0008] wherein Y.sup.- is an anionic group at physiological pH; Q
is a carrier molecule; X.sup.+ is a cationic group; and n is an
integer selected such that the biodistribution of the therapeutic
compound for an intended target site is not prevented while
maintaining activity of the therapeutic compound, or a
pharmaceutically acceptable salt or ester thereof, such that the
glycosaminoglycan-associated molecular interaction is modulated and
the condition is treated. These methods can be used therapeutically
to treat a subject, e.g., afflicted with a pathogen, or can be used
prophylactically in a subject susceptible to pathogens.
[0009] In another embodiment, the therapeutic compound has at least
one anionic group covalently attached to a carrier molecule. In
another embodiment, the anionic group covalently attached to the
carrier molecule is a sulfonate group. Accordingly, the therapeutic
compound can have the formula: QSO.sub.3.sup.-X.sup.+].sub.n
(II)
[0010] wherein Q is a carrier molecule; X.sup.+ is a cationic
group; and n is an integer. In another embodiment, the anionic
group is a sulfate group. Accordingly, the therapeutic compound can
have the formula: QOSO.sub.3.sup.-X.sup.+].sub.n (III) wherein Q is
a carrier molecule; X.sup.+ is a cationic group; and n is an
integer. Carrier molecules which can be used include carbohydrates,
polymers, peptides, peptide derivatives, aliphatic groups,
alicyclic groups, heterocyclic groups, aromatic groups and
combinations thereof.
[0011] The invention also provides a method for modulating
interactions between an infectious agent and a GAG in a subject.
The method includes administering to the subject a therapeutically
effective amount of a therapeutic compound having the formula:
QY.sup.-X.sup.+].sub.n (I)
[0012] wherein Y.sup.- is an anionic group at physiological pH; Q
is a carrier molecule; X.sup.+ is a cationic group; and n is an
integer selected such that the biodistribution of the therapeutic
compound for an intended target site is not prevented while
maintaining activity of the therapeutic compound, or a
pharmaceutically acceptable salt or ester thereof.
[0013] In another aspect, methods and therapeutic compositions are
provided herein for treating a subject afflicted with a disease,
e.g., acute rheumatic fever and poststreptococcal
glomerulonephritis, caused by infection by bacteria such as
Streptococcus pyogenes, Chlamydia trachomatis, Staphylococcus
aureus, Pseudomonas aeruginosa, Legionella pneumophila, Bordetella
pertussis, and Mycoplasma pneumoniae, such that the subject
afflicted with the disease is treated. The methods include
administering to a subject a therapeutically effective amount of a
therapeutic compound of formula (I) for treating the infection. The
therapeutic compound is not carrageenan, pentosan polysulfate,
fucoidan, dextran sulfate, heparin, heparan sulfate or dermatan
sulfate.
[0014] In yet another aspect, the invention provides methods and
therapeutic compositions for treating a subject afflicted with a
disease caused by infection of viri via such as Cytomegalovirus
(CMV) and Herpes simplex (HSV-1 and HSV-2), such that the subject
afflicted with the disease is treated. The methods include
administering to a subject a therapeutically effective amount of a
therapeutic compound of formula (I) for treating the disease. The
therapeutic compound is not a chondroitin sulfate.
[0015] In yet a further aspect a packaged pharmaceutical
composition for treating a condition related to a
glycosaminoglycan-associated molecular interaction or for
modulating a GAG-associated molecular interaction, e.g., between a
GAG and an infectious agent, is described herein. The packaged
composition includes a container holding a therapeutically
effective amount of a pharmaceutical composition for treating the
condition related to a glycosaminoglycan-associated molecular
interaction in a subject. Alternatively, the packaged composition
includes a container holding a therapeutically effective amount of
a pharmaceutical composition for modulating a GAG-associated
molecular interaction. The pharmaceutical composition includes at
least one therapeutic compound having the formula:
QY.sup.-X.sup.+].sub.n (I)
[0016] wherein Y.sup.- is an anionic group at physiological pH; Q
is a carrier molecule; X.sup.+ is a cationic group; and n is an
integer selected such that the biodistribution of the therapeutic
compound for an intended target site is not prevented while
maintaining activity of the therapeutic compound, or a
pharmaceutically acceptable salt or ester thereof. Instructions for
using the pharmaceutical composition for treatment of the condition
related to a glycosaminoglycan-associated molecular interaction or
for modulating the GAG-associated molecular interaction are
included in the packaged pharmaceutical composition.
[0017] The invention further provides pharmaceutical compositions
for treating a condition related to a glycosaminoglycan-associated
molecular interaction in a subject. Alternatively, the invention
provides pharmaceutical compositions for modulating a
GAG-associated molecular interaction in a subject. The
pharmaceutical compositions include a therapeutically effective
amount of a therapeutic compound of the invention, as described
supra, and a pharmaceutically acceptable carrier.
[0018] In further embodiments, the therapeutic compound has at
least one anionic group covalently attached to a carrier molecule.
In yet another embodiment, the anionic group covalently attached to
the carrier molecule is a sulfonate group. Accordingly, the
therapeutic compound can have the formula:
QSO.sub.3.sup.-X.sup.+].sub.n (II)
[0019] wherein Q is a carrier molecule; X.sup.+ is a cationic
group; and n is an integer. In another embodiment, the anionic
group is a sulfate group. Accordingly, the therapeutic compound can
have the formula: QOSO.sub.3.sup.-X.sup.+].sub.n (III)
[0020] wherein Q is a carrier molecule; X.sup.+ is a cationic
group; and n is an integer.
BRIEF DESCRIPTION OF THE DRAWING
[0021] FIGS. 1-14 depict the chemical structures of compounds
described in the specification.
[0022] FIGS. 15-28 illustrate the efficacy of compounds of the
invention in inhibiting binding of certain compounds, e.g., Rantes,
IL-8, to heparan-coated wells. The compounds referenced in the
drawings are: 1) 3-amino-1-propanesulfonic acid, sodium salt; 2)
trisodium phosphonoformate; 3) methylene diphosphonic acid; 4)
trehalose octasulfate, octasodium salt; 5)
trans-4-hydroxy-L-proline-4-sulfate, disodium salt; 6)
nitrilo(methylene) triphosphonic acid; 7) poly(vinylsulfonate),
sodium salt (PVS501, Aldrich); 8)
3-[-2-6-methoxy-1,2,3,4-tetrahydroisoquinolinyl)]-1-propanesulfonic
add; 9) 3-phosphonopropanesulfonic acid, trisodium salt; 10);
4,5-dihydroxy-1,3,benzenedisulfonic acid, sodium salt; 11)
3-cyclohexylamino-1-propanesulfonic acid; 12) O-phospho-L-serine;
and 13) 2-thiopheneboronic acid.
DETAILED DESCRIPTION OF THE INVENTION
[0023] The features and other details of the invention will now be
more particularly described with reference to the accompanying
drawings and pointed out in the claims. It will be understood that
particular embodiments described herein are shown by way of
illustration and not as limitations of the invention. The principal
features of this invention can be employed in various embodiments
without departing from the scope of the invention. All parts and
percentages are by weight unless otherwise specified.
[0024] The invention provides methods and compositions which are
useful in the treatment of conditions related to glycosaminoglycan
(GAG)-associated molecular interactions. In one embodiment, the
invention provides a method for treating a condition related to a
glycosaminoglycan-associated molecular interaction in a subject.
The method includes administering to the subject, a therapeutically
effective amount of a therapeutic compound having the formula:
QY.sup.-X.sup.+].sub.n (I)
[0025] wherein Y.sup.- is an anionic group at physiological pH; Q
is a carrier molecule; X.sup.+ is a cationic group; and n is an
integer selected such that the biodistribution of the therapeutic
compound for an intended target site is not prevented while
maintaining activity of the therapeutic compound, or a
pharmaceutically acceptable salt of ester thereof. The methods of
the invention can be used therapeutically to treat a subject
afflicted by a pathogen or can be used prophylactically in a
subject susceptible to pathogens. The methods of the invention are
based, at least in part, on inhibiting, eradicating, or preventing
interaction between the cell membrane surface and the pathogen.
[0026] The language "treating a condition related to a
glycosaminoglycan (GAG)-associated molecular interaction" and
"treatment of a condition related to a glycosaminoglycan-associated
molecular interaction" is intended to include changes in a
condition related to a glycosaminoglycan-associated molecular
interaction, as described infra, such that physiological symptoms
in a subject can be significantly diminished or minimized. The
language also includes control, prevention, relief, or inhibition
of physiological symptoms or effects attributed to a disease state
associated with glycosaminoglycan-associated molecular
interactions. In one preferred embodiment, the control of the
glycosaminoglycan-associated molecular interaction or condition
related thereto is such that the glycosaminoglycan-associated
molecular interaction or condition related thereto is eradicated.
In another preferred embodiment, the control is selective such that
a particular targeted glycosaminoglycan-associated molecular
interaction, e.g., with a pathogen, is controlled while other cells
and physiological flora which are not detrimental to the subject
are allowed to remain substantially uncontrolled or substantially
unaffected, e.g., lymphocytes, red blood cells, white blood cells,
platelets, growth factors, etc.
[0027] The term "pathogen" is art recognized and is intended to
include disease producing agents, such as organisms, e.g.,
microorganisms, capable of causing disease in a subject, e.g., a
mammal, including, for example, bacteria, viruses, prions and
fungi.
[0028] As used herein, "glycosaminoglycan (GAG)-associated
molecular interaction" is intended to include the binding of a GAG
to, for example, a cell surface, secreted, or extracellular
protein. This term also includes any subsequent results of such
protein binding such as, for example, delayed proteolytic
degradation or denaturing, changes in protein conformation (which
may, for example, lead to alterations of biological activity), or
catalysis of a reaction between two different proteins bound to the
same or different GAGs on the same or different proteoglycans. Also
included is the ability of certain GAGs, e.g., heparin sulfate, to
modulate the interaction of a protein to another GAG, for example,
FGF-2 (basic fibroblast growth factor) to its GAG cell
receptor.
[0029] Other GAG-associated molecular interactions include specific
interactions between specific compounds and factors. Non-limiting
examples include polypeptide growth factors (e.g., FGFs1-9, PDGF,
HGF, VEGF, TGF-.beta., IL-3); extracellular matrix components
(e.g., laminins, fibronectins; thrombospondins, tenascins,
collagens, VonWirebrand's factor); proteases and anti-proteases
(e.g., thrombin, TPA, UPA, dotting factors IX and X, PAI-1);
cell-adhesion molecules (e.g., N-CAM, LI, myelin-associated
glycoprotein); proteins involved in lipoprotein metabolism (e.g.,
APO-B, APO-E, lipoprotein lipase); cell-cell adhesion molecules
(e.g., N-CAM, myelin-associated glycoprotein, selectins, pecam);
angiogenin; lactoferrin; viral proteins (e.g., proteins from HIV,
herpes complex) and other compounds which bind to GAG. The
definition is intended to include the result of the binding of
these factors to the GAG. For example, the binding of polypeptide
growth factor to a GAG can result in cell proliferation,
angiogenesis, inflammation, cancer, and other biologically
important responses.
[0030] Also, the term "glycosaminoglycan (GAG)-associated molecular
interaction" includes microbial "interactions" with GAGs which may,
for example, lead to invasion of a host cell by a microorganism. It
includes the binding of adhesius or other microbial proteins to GAG
and the results thereafter. Some examples of such adhesius are: the
filamentous hemagluggtinin of Bordetella pertussis, gP120 of HIV,
gpB and pgC of HSV, etc. It also includes interactions where the
GAG functions as a bridge between the microbial organism and the
host cell, e.g., in chlamydia trachomatis, heparin sulfate binds to
both C. trachomatis to host cell receptors catalyzing an
interaction between the two. Also included in the definition is any
interaction between a microbial organism and a cell which is
mediated through the GAG.
[0031] The term "glycosaminoglycan (GAG)-associated molecular
interaction" is further intended to include disease states or
conditions caused by or associated with one or more pathogens which
interact with extracellular membrane components, e.g.,
glycosaminoglycans, often found on host cell surfaces. In one
embodiment, the disease state includes, for example, those diseases
which afflict a subject by associating with or interfering with
glycosaminoglycans found within the subject. In a preferred
embodiment, the term "glycosaminoglycan-associated molecular
interaction" does not include amyloidosis. In another preferred
embodiment, the term "glycosaminoglycan-associated molecular
interaction" does not include interactions between an amyloidogenic
protein and a constituent of basement membrane to inhibit amyloid
deposition. In yet another preferred embodiment, the term
"glycosaminoglycan" does not include a constituent of basement
membrane, e.g., heparan sulfate proteoglycan. In yet another
preferred embodiment, the term "glycosaminoglycan" does not include
sulfated GAGs, e.g., heparan sulfate. Presently unknown conditions
related to glycosaminoglycan-associated molecular interactions that
may be discovered in the future are encompassed, since their
characterization as conditions related to
glycosaminoglycan-associated molecular interactions will be readily
determinable by persons skilled in the art.
[0032] Conditions related to glycosaminoglycan-associated molecular
interactions include, for example, certain bacteria such as
Streptococcus pyogenes, associated with acute rheumatic fever and
poststreptococcal glomerulonephritis, Chlamydia trachomatis,
Staphylococcus aureus (cystic fibrosis), Bordetella pertussis
(whooping cough) and Mycoplasma pneumoniae. For example,
Streptococcus pyogenes surfaces bind fibronectin, laminin,
fibrinogen, nonspecific immunoglobulins A and G,
.alpha.2-macroglobulin, .beta.2-microglobulin and albumin.
Infection by Chlamydia trachomatis is facilitated by binding of a
heparan sulfate-like GAG present on the surface of chlamydia, to a
heparan sulfate receptor on the target cell.
[0033] Additionally, conditions related to
glycosaminoglycan-associated molecular interactions include certain
types of viri, such as Herpesviridae, which are believed to be
associated with HSPG during the infectious process. These viri
appear to interact with a cell's surface through GAGs found on the
proteoglycans of the cell plasma membrane. These GAGs are similar
to heparin. Cytomegalovirus (CMV), HIV and Herpes simplex (HSV-1
and HSV-2) are examples of which are believed to infect cells via
cell surface GAGs.
[0034] In one aspect, the present invention pertains to methods for
modulating a glycosaminoglycan-associated molecular interaction,
e.g., between an infectious agent and a GAG, in a subject. The
methods include administering to the subject a therapeutically
effective amount of a therapeutic compound. The therapeutic
compound has the formula: QY.sup.-X.sup.+].sub.n (I)
[0035] wherein Y.sup.- is an anionic group at physiological pH; Q
is a carrier molecule; X.sup.+ is a cationic group; and n is an
integer selected such that the biodistribution of the therapeutic
compound for an intended target site is not prevented while
maintaining activity of the therapeutic compound, or a
pharmaceutically acceptable salt or ester thereof.
[0036] The term "infectious agent" is intended to include those
pathogens which are associated with disease states caused by
bacteria, viri or prions. The term is also intended to include
those extracellular components, e.g., proteins, etc., which are
secreted, produced, or otherwise discharged by a pathogen, thereby
causing the subject to be afflicted with a disease state associated
with the infectious agent. Those disease states associated with
infectious agents include Streptococcus pyogenes, Chlamydia
trachomatis, Staphylococcus aureus, Bordetella pertussis,
Mycoplasma pneumoniae, Herpesvzridae, e.g., herpes simplex. The
term infectious agent is also intended to encompass presently
unknown infectious agents that may be discovered in the future,
since their characterization as a infectious agents will be readily
determinable by persons skilled in the art.
[0037] In an embodiment, therapeutic compounds which comprise at
least one sulfate group covalently attached to a carrier molecule,
or pharmaceutically acceptable salt thereof are used to treat a
condition related to a glycosaminoglycan (GAG)-associated molecular
interaction or an infectious agent. In particular, the therapeutic
compounds of the invention comprise at least one sulfate group or a
functional equivalent thereof, for example a sulfonic acid group or
other functionally equivalent anionic group, linked to a carrier
molecule. In addition to functioning as a carrier for the anionic
functionality, the carrier molecule can enable the compound to
traverse biological membranes and to be biodistributed without
excessive or premature metabolism. Moreover, when multiple anionic
functionalities are present on a carrier molecule, the carrier
molecule serves to space the anionic groups in a correct geometric
separation.
[0038] In one embodiment, when the condition related to a
glycosaminoglycan-associated molecular interaction is associated
with or caused by the bacteria Chlamydia trachomatis, the
therapeutic compound is not sulfated polysaccharides kapp and
lambda, iota carrageenans C-1263, C-3889 and C-4014, pentosan
polysulfate (P-8275), fucodian (F-5631), dextran sulfate (D-6001),
heparin (H-3393), heparan sulfate (H-7641) or dermatan sulfate
(chondroitin sulfate A and B).
[0039] In another embodiment, when the condition related to a
glycosaminoglycan-associated molecular interaction is associated
with or caused by cytomegalovirus, the therapeutic compound is not
pentosan polysulfate, dextran sulfate, heparin, copolymers of
acrylic acid and vinylalcohol sulfate, .alpha.-cyclodextrin
hexasulfate and .alpha.-cyclodextrin dodecasulfate.
[0040] In yet another embodiment, when the condition related to a
glycosaminoglycan-associated molecular interaction is associated
with the virus Herpesviridae, the therapeutic agent is not
chondroitin sulfate A, B or C.
[0041] In one embodiment, the method of the invention includes
administering to the subject an effective amount of a therapeutic
compound which has at least one anionic group covalently attached
to a carrier molecule. The therapeutic compound is capable of
treating a condition related to a glycosaminoglycan-associated
molecular interaction or an infectious agent. The therapeutic
compound can have the formula: QY.sup.-X.sup.+].sub.n (I)
[0042] wherein Y.sup.- is an anionic group at physiological pH; Q
is a carrier molecule; X.sup.+ is a cationic group; and n is an
integer. The number of anionic groups ("n") is selected such that
the biodistribution of the compound for an intended target site is
not prevented while maintaining activity of the compound. For
example, the number of anionic groups is not so great as to inhibit
traversal of an anatomical barrier, such as a cell membrane, or
entry across a physiological barrier, such as the blood-brain
barrier, in situations where such properties are desired. In one
embodiment, n is an integer between 1 and 10. In another
embodiment, n is an integer between 3 and 8.
[0043] An anionic group of a therapeutic compound of the invention
is a negatively charged moiety that, when attached to a carrier
molecule, can inhibit an interaction between a bacteria, a virus or
an infectious agent and a cell membrane. The anionic group is
desirably negatively charged at physiological pH. Preferably, the
anionic therapeutic compound mimics the structure of a sulfated
proteoglycan, i.e., is a sulfated compound or a functional
equivalent thereof. "Functional equivalents" of sulfates are
intended to include compounds such as sulfamates as well as
bioisosteres. Bioisosteres encompass both classical bioisosteric
equivalents and non-classical bioisosteric equivalents. Classical
and nonclassical bioisosteres of sulfate groups are known in the
art (see, e.g., Silverman, R. B. The Organic Chemistry of Drug
Design and Drug Action, Academic Press, Inc. San Diego, Calif.,
1992, pp. 19-23). Accordingly, a therapeutic compound of the
invention can comprise at least one anionic group including
sulfonates, sulfates, sulfamates, phosphonates, phosphates,
carboxylates, and heterocyclic groups of the following formulae:
##STR1##
[0044] Depending on the carrier molecule, more than one anionic
group can be attached thereto. When more than one anionic group is
attached to a carrier molecule, the multiple anionic groups can be
the same structural group (e.g., all sulfonates) or, alternatively,
a combination of different anionic groups can be used (e.g.,
sulfonates and sulfates, etc.).
[0045] A therapeutic compound of the invention typically further
comprises a counter cation (i.e., X.sup.+ in formula (I):
QY.sup.-X.sup.+].sub.n). Cationic groups include positively charged
atoms and moieties. If the cationic group is hydrogen, H.sup.+,
then the compound is considered an acid, e.g., ethanesulfonic acid.
If hydrogen is replaced by a metal or its equivalent, the compound
is a salt of the acid. Pharmaceutically acceptable salts of the
therapeutic compound are within the scope of the invention. For
example, X.sup.+ can be a pharmaceutically acceptable alkali metal,
alkaline earth, higher valency cation (e.g., aluminum salt),
polycationic counter ion or ammonium. A preferred pharmaceutically
acceptable salt is a sodium salt but other salts are also
contemplated within their pharmaceutically acceptable range.
[0046] Within the therapeutic compound, the anionic group(s) is
covalently attached to a carrier molecule. Suitable carrier
molecules include carbohydrates, polymers, peptides, peptide
derivatives, aliphatic groups, alicyclic groups, heterocyclic
groups, aromatic groups or combinations thereof. A carrier molecule
can be substituted, e.g. with one or more amino, nitro, halogen,
thiol or hydroxy groups.
[0047] As used herein, the term "carbohydrate" is intended to
include substituted and unsubstituted mono-, oligo-, and
polysaccharides. Monosaccharides are simple sugars usually of the
formula C.sub.6H.sub.12O.sub.6 that can be combined to form
oligosaccharides or polysaccharides. Monosaccharides include
enantiomers and both the D and L stereoisomers of monosaccharides.
Carbohydrates can have multiple anionic groups attached to each
monosaccharide moiety. For example, in sucrose octasulfate, four
sulfate groups are attached to each of the two monosaccharide
moieties.
[0048] As used herein, the term "polymer" is intended to include
molecules formed by the chemical union of two or more combining
subunits called monomers. Monomers are molecules or compounds which
usually contain carbon and are of relatively low molecular weight
and simple structure. A monomer can be converted to a polymer by
combination with itself or other similar molecules or compounds. A
polymer may be composed of a single identical repeating subunit or
multiple different repeating subunits (copolymers). Polymers within
the scope of this invention include substituted and unsubstituted
vinyl, acryl, styrene and carbohydrate-derived polymers and
copolymers and salts thereof. In one embodiment, the polymer has a
molecular weight of approximately 800-1000 Daltons. Examples of
polymers with suitable covalently attached anionic groups (e.g.,
sulfonates or sulfates) include
poly(2-acrylamido-2-methyl-1-propanesulfonic acid);
poly(2-acrylamido-2-methyl-1-propanesulfonic
acid-co-acrylonitrile);
poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-styrene);
poly(vinylsulfonic acid); poly(sodium 4-styrenesulfonic acid); and
sulfates and sulfonates derived from: poly(acrylic acid);
poly(methyl acrylate); poly(methyl methacrylate); and poly(vinyl
alcohol); and pharmaceutically acceptable salts thereof. Examples
of carbohydrate-derived polymers with suitable covalently attached
anionic groups include those of the formula: ##STR2##
[0049] wherein R is SO.sub.3.sup.- or OSO.sub.3.sup.-; and
pharmaceutically acceptable salts thereof.
[0050] Peptides and peptide derivatives can also act as carrier
molecules. The term "peptide" includes two or more amino acids
covalently attached through a peptide bond. Amino acids which can
be used in peptide carrier molecules include those naturally
occurring amino adds found in proteins such as glycine, alanine,
valine, cysteine, leucine, isoleucine, serine, threonine,
methionine, glutamic acid, aspartic acid, glutamine, asparagine,
lysine, arginine, proline, histidine, phenylalanine, tyrosine, and
tryptophan. The term amino acid further includes analogs,
derivatives and congeners of naturally occurring amino acids, one
or more of which can be present in a peptide derivative. For
example, amino add analogs can have lengthened or shortened side
chains or variant side chains with appropriate functional
groups.
[0051] Also included are the D and L stereoisomers of an amino acid
when the structure of the amino acid admits of stereoisomeric
forms. The term "peptide derivative" further includes compounds
which contain molecules which mimic a peptide backbone but are not
amino acids (so-called peptidomimetics), such as benzodiazepine
molecules (see e.g. James, G. L. et al. (1993) Science
260:1937-1942). The anionic groups can be attached to a peptide or
peptide derivative through a functional group on the side chain of
certain amino acids or other suitable functional group. For
example, a sulfate or sulfonate group can be attached through the
hydroxy side chain of a serine residue. Accordingly, in one
embodiment, the peptide comprises four amino acids and anionic
groups (e.g., sulfonates) are attached to the first, second and
fourth amino acid. For example, the peptide can be Ser-Ser-Y-Ser,
wherein an anionic group is attached to the side chain of each
serine residue and Y is any amino acid. In addition to peptides and
peptide derivatives, single amino adds can be used as carriers in
the therapeutic compounds of the invention. For example, cysteic
acid, the sulfonate derivative of cysteine, can be used.
[0052] The term "aliphatic group" is intended to include organic
compounds characterized by straight or branched chains, typically
having between 1 and 22 carbon atoms. Aliphatic groups include
alkyl groups, alkenyl groups and alkynyl groups. In complex
structures, the chains can be branched or cross-linked. Alkyl
groups include saturated hydrocarbons having one or more carbon
atoms, including straight-chain alkyl groups and branched-chain
alkyl groups. Such hydrocarbon moieties may be substituted on one
or more carbons with, for example, a halogen, a hydroxyl, a thiol,
an amino, an alkoxy, an alkylcarboxy, an alkylthio, or a nitro
group. Unless the number of carbons is otherwise specified, "lower
aliphatic" as used herein means an aliphatic group, as defined
above (e.g., lower alkyl, lower alkenyl, lower alkynyl), but having
from one to six carbon atoms. Representative of such lower
aliphatic groups, e.g., lower alkyl groups, are methyl, ethyl,
n-propyl, isopropyl, 2-chloropropyl, n-butyl, sec-butyl,
2-aminobutyl, isobutyl, tert-butyl, 3-thiopentyl, and the like. As
used herein, the term "amino" means --NH.sub.2; the term "nitro"
means --NO.sub.2; the term "halogen" designates --F, --Cl, --Br or
--I; the term "thiol" means SH; and the term "hydroxyl" means --OH.
Thus, the term "alkylamino" as used herein means an alkyl group, as
defined above, having an amino group attached thereto. The term
"alkylthio" refers to an alkyl group, as defined above, having a
sulfhydryl group attached thereto. The term "alkylcarboxyl" as used
herein means an alkyl group, as defined above, having a carboxyl
group attached thereto. The term "alkoxy" as used herein means an
alkyl group, as defined above, having an oxygen atom, attached
thereto. Representative alkoxy groups include methoxy, ethoxy,
propoxy, tert-butoxy and the like. The terms "alkenyl" and
"alkynyl" refer to unsaturated aliphatic groups analogous to
alkyls, but which contain at least one double or triple bond
respectively.
[0053] The term "alicyclic group" is intended to include dosed ring
structures of three or more carbon atoms. Alicyclic groups include
cycloparaffins or naphthenes which are saturated cyclic
hydrocarbons, cycloolefins which are unsaturated with two or more
double bonds, and cycloacetylenes which have a triple bond. They do
not include aromatic groups. Examples of cycloparaffins include
cyclopropane, cyclohexane, and cyclopentane. Examples of
cycloolefins include cyclopentadiene and cyclooctatetraene.
Alicyclic groups also include fused ring structures and substituted
alicyclic groups such as alkyl substituted alicyclic groups. In the
instance of the alicyclics such substituents can further comprise a
lower alkyl, a lower alkenyl, a lower alkoxy, a lower alkylthio, a
lower alkylamino, a lower alkylcarboxyl, a nitro, a hydroxyl,
--CF.sub.3, --CN, or the like.
[0054] The term "heterocyclic group" is intended to include closed
ring structures in which one or more of the atoms in the ring is an
element other than carbon, for example, nitrogen, or oxygen.
Heterocyclic groups can be saturated or unsaturated and
heterocyclic groups such as pyrrole and furan can have aromatic
character. They include fused ring structures such as quinoline and
isoquinoline. Other examples of heterocyclic groups include
pyridine and purine. Heterocyclic groups can also be substituted at
one or more constituent atoms with, for example, a halogen, a lower
alkyl, a lower alkenyl, a lower alkoxy, a lower alkylthio, a lower
alkylamino, a lower alkylcarboxyl, a nitro, a hydroxyl, --CF.sub.3,
--CN, or the like.
[0055] The term "aromatic group" is intended to include unsaturated
cyclic hydrocarbons containing one or more rings. Aromatic groups
include 5- and 6-membered single-ring groups which may include from
zero to four heteroatoms, for example, benzene, pyrrole, furan,
thiophene, imidazole, oxazole, thiazole, triazole, pyrazole,
pyridine, pyrazine, pyridazine and pyrimidine, and the like. The
aromatic ring may be substituted at one or more ring positions
with, for example, a halogen, a lower alkyl, a lower alkenyl, a
lower alkoxy, a lower alkylthio, a lower alkylamino, a lower
alkylcarboxyl, a nitro, a hydroxyl, --CF.sub.3, --CN, or the
like.
[0056] The therapeutic compound of the invention can be
administered in a pharmaceutically acceptable carrier. As used
herein "pharmaceutically acceptable carrier" includes any and all
solvents, dispersion media, coatings, antibacterial and antifungal
agents, isotonic and absorption delaying agents, and the like which
are compatible with the activity of the compound and are
physiologically acceptable to the subject. An example of a
pharmaceutically acceptable carrier is buffered normal saline (0.15
molar NaCl). The use of such media and agents for pharmaceutically
active substances is well known in the art. Except insofar as any
conventional media or agent is incompatible with the therapeutic
compound, use thereof in the compositions suitable for
pharmaceutical administration is contemplated. Supplementary active
compounds can also be incorporated into the compositions.
[0057] In an embodiment of the method of the invention, the
therapeutic compound administered to the subject is comprised of at
least one sulfonate group covalently attached to a carrier
molecule, or a pharmaceutically acceptable salt thereof.
Accordingly, the therapeutic compound can have the formula:
QSO.sub.3.sup.-X.sup.+].sub.n (II)
[0058] wherein Q is a carrier molecule; X.sup.+ is a cationic
group; and n is an integer. Suitable carrier molecules and cationic
groups are those described hereinbefore. The number of sulfonate
groups ("n") is selected such that the biodistribution of the
compound for an intended target site is not prevented while
maintaining activity of the compound as discussed earlier. In one
embodiment, n is an integer between 1 and 10. In another
embodiment, n is an integer between 3 and 8. As described earlier,
therapeutic compounds with multiple sulfonate groups can have the
sulfonate groups spaced such that the compound interacts optimally
with a receptor site on the cell membrane.
[0059] In certain embodiments, the carrier molecule for a
sulfonate(s) is a lower aliphatic group (e.g., a lower alkyl, lower
alkenyl or lower alkynyl), a heterocyclic group, a disaccharide, a
polymer or a peptide or peptide derivative. Furthermore, the
carrier can be substituted, e.g. with one or more amino, nitro,
halogen, thiol or hydroxy groups. In certain embodiments, the
carrier molecule for a sulfonate(s) is an aromatic group.
[0060] Particularly suitable therapeutic compounds include
1,3-propanedisulfonic acid, 3-amino-1-propanesulfonic acid,
3-dimethylamino-1-propanesulfonic acid sodium salt,
2-(3-sulfopropyl)-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole, sodium
salt,
3-[2-(6-methoxy-1,2,3,4-tetrahydroisoquinolinyl)]-1-propanesulfonic
acid, 3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(3-hydroxy-1-propyl)amino-1-propanesulfonic acid,
(-)3-[(R)-2-hydroxy-1-propyl]amino-1-propanesulfonic acid,
3-(4-hydroxy-1-butyl)amino-1-propanesulfonic acid,
3-(5-hydroxy-1-pentyl)amino-1-propanesulfonic acid,
3-(6-hydroxy-1-hexyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-hexylarnino-1-propanesulfonic acid,
3-undecylamino-1-propanesulfonic acid, and
3-octadecylamino-1-propanesulfonic acid, and pharmaceutically
acceptable salts or esters thereof.
[0061] Examples of sulfonated polymeric therapeutic compounds
include poly(2-acrylamido-2-methyl-1-propanesulfonic acid);
poly(2-acrylamido-2-methyl-1-propanesulfonic
acid-co-acrylonitrile);
poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-styrene);
poly(vinylsulfonic acid); poly(sodium 4-styrenesulfonic acid); a
sulfonic acid derivative of poly(acrylic acid); a sulfonic acid
derivative of poly(methyl acrylate); a sulfonic acid derivative of
poly(methyl methacrylate); and a sulfonate derivative of poly(vinyl
alcohol); and pharmaceutically acceptable salts thereof.
[0062] A suitable sulfonated polymer is poly(vinylsulfonic acid)
(PVS) or a pharmaceutically acceptable salt thereof, preferably the
sodium salt thereof. In one embodiment, PVS having a molecular
weight of about 800-1000 Daltons is used. PVS may be used as a
mixture of stereoisomers or as a single active isomer.
[0063] A suitable sulfonated disaccharide is a fully or partially
sulfonated sucrose, or pharmaceutically acceptable salt thereof,
such as sucrose octasulfonate. Other sulfonated saccharides include
5-deoxy-1,2-O-isopropylidene-.alpha.-D-xylofuranose-5-sulfonic acid
(XXIII, shown as the sodium salt).
[0064] Suitable lower aliphatic sulfonated compounds for use in the
invention include ethanesulfonic acid; 2-aminoethanesulfonic acid
(taurine); cysteic acid (3-sulfoalanine or
.alpha.-amino-.beta.-sulfopropionic acid); 1-propanesulfonic acid;
1,2-ethanedisulfonic acid; 1,4-butanedisulfonic acid;
1,5-pentanedisulfonic acid; and 4-hydroxybutane-1-sulfonic acid
(VIII, shown as the sodium salt); and pharmaceutically acceptable
salts thereof. Other aliphatic sulfonated compounds contemplated
for use in the invention include 1-butanesulfonic acid (XLVII,
shown as the sodium salt), 2-propanesulfonic acid (XLIX, shown as
the sodium salt), 3-pentanesulfonic acid (L, shown as the sodium
salt), 4-heptanesulfonic acid (LII, shown as the sodium salt),
1-decanesulfonic acid (XLVIII, shown as the sodium salt); and
pharmaceutically acceptable salts thereof. Sulfonated substituted
aliphatic compounds contemplated for use in the invention include
3-amino-1-propanesulfonic acid (XXII, shown as the sodium salt),
3-hydroxypropanesulfonic acid sulfate (XXXV, shown as the disodium
salt), 1,7-dihydroxy-4-heptanesulfonic acid (LIII, shown as the
sodium salt); and pharmaceutically acceptable salts thereof. Yet
other sulfonated compounds contemplated for use in the invention
include 2-[(4-pyridinyl)amido]ethanesulfonic add (LIV, depicted as
the sodium salt), and pharmaceutically acceptable salts
thereof.
[0065] Suitable heterocyclic sulfonated compounds include
3-(N-morpholino)propanesulfonic acid; and
tetrahydrothiophene-1,1-dioxide-3,4-disulfonic acid; and
pharmaceutically acceptable salts thereof.
[0066] Aromatic sulfonated compounds include 1,3-benzenedisulfonic
acid (XXXVI, shown as the disodium salt),
2,5-dimethoxy-1,4-benzenedisulfonic acid (depicted as the disodium
salt, XXXVII, or the dipotassium salt, XXXIX),
4-amino-3-hydroxy-1-naphthalenesulfonic add (XLIII),
3,4-diamino-1-naphthalenesulfonic acid (XLIV); and pharmaceutically
acceptable salts thereof.
[0067] In another embodiment of the method of the invention, the
therapeutic compound administered to the subject is comprised of at
least one sulfate group covalently attached to a carrier molecule,
or a pharmaceutically acceptable salt thereof. Accordingly, the
therapeutic compound can have the formula:
QOSO.sub.3.sup.-X.sup.+].sub.n (III)
[0068] wherein Q is a carrier molecule; X.sup.+ is a cationic
group; and n is an integer. Suitable carrier molecules and cationic
groups are those described hereinbefore. The number of sulfate
groups ("n") is selected such that the biodistribution of the
compound for an intended target site is not prevented while
maintaining activity of the compound as discussed earlier. In one
embodiment, n is an integer between 1 and 10. In another
embodiment, n is an integer between 3 and 8. As described earlier,
therapeutic compounds with multiple sulfate groups can have the
sulfate groups spaced such that the compound interacts optimally
with a bacteria, virus or an infectious agent or a cell
membrane.
[0069] In certain embodiments, the carrier molecule for a
sulfate(s) is a lower aliphatic group (e.g., a lower alkyl, lower
alkenyl or lower alkynyl), an aromatic group, a disaccharide, a
polymer or a peptide or peptide derivative. Furthermore, the
carrier can be substituted, e.g. with one or more amino, nitro,
halogen, thiol or hydroxy groups.
[0070] Examples of sulfated polymeric therapeutic compounds include
poly(2-acrylamido-2-methyl-propyl sulfuric acid);
poly(2-acrylamido-2-methyl-propyl sulfuric acid-co-acrylonitrile);
poly(2-acrylamido-2-methyl-propyl sulfuric acid-co-styrene);
poly(vinylsulfuric acid); poly(sodium 4-styrenesulfate); a sulfate
derivative of poly(acrylic acid); a sulfate derivative of
poly(methyl acrylate); a sulfate derivative of poly(methyl
methacrylate); and a sulfate derivative of poly(vinyl alcohol); and
pharmaceutically acceptable salts thereof.
[0071] A suitable sulfated polymer is poly(vinylsulfuric acid) or
pharmaceutically acceptable salt thereof. A suitable sulfated
disaccharide is sucrose octasulfate or pharmaceutically acceptable
salt thereof. Other contemplated sulfated saccharides include the
add form of methyl-.alpha.-D-glucopyranoside 2,3-disulfate (XVI),
methyl 4,6-O-benzylidene-.alpha.-D-glucopyranoside 2,3-disulfate
(XVII), 2,3,4,3',4'-sucrose pentasulfate (XXXIII),
1,3:4,6-di-O-benzylidene-D-mannitol 2,5-disulfate (XLI), D-mannitol
2,5-disulfate (XLII), 2,5-di-O-benzyl-D-mannitol tetrasulfate
(XLV); and pharmaceutically acceptable salts thereof.
[0072] Suitable lower aliphatic sulfated compounds for use in the
invention include ethyl sulfuric acid; 2-aminoethan-1-ol sulfuric
acid; 1-propanol sulfuric acid; 1,2-ethanediol disulfuric add;
1,3-propanediol disulfuric acid; 1,4-butanediol disulfuric acid;
1,5-pentanediol disulfuric acid; and 1,4-butanediol monosulfuric
acid; and pharmaceutically acceptable salts thereof. Other sulfated
aliphatic compounds contemplated for use in the invention include
the acid form of 1,3-cydohexanediol disulfate (XL),
1,3,5-heptanetriol trisulfate (XIX),
2-hydroxymethyl-1,3-propanediol trisulfate (XX),
2-hydroxymethyl-2-methyl-1,3-propanediol trisulfate (XXI),
1,3,5,7-heptanetetraol tetrasulfate (XLVI), 1,3,5,7,9-nonane
pentasulfate (LI); and pharmaceutically acceptable salts thereof.
Other sulfated compounds contemplated for use in the invention
include the acid form of 2-amino-2-hydroxymethyl-1,3-propanediol
trisulfate (XXIV), 2-benzyloxy-1,3-propanediol disulfate (XXIX),
3-hydroxypropylsulfamic acid sulfate (XXX)2,2'-iminoethanol
disulfate (XXXI), N,N-bis(2-hydroxyethyl)sulfamic acid disulfate
(XXXII); and pharmaceutically acceptable salts thereof.
[0073] Suitable heterocyclic sulfated compounds include
3-(N-morpholino) propanesulfuric acid; and
tetrahydrothiophene-1,1-dioxide-3,4-diol disulfuric acid; and
pharmaceutically acceptable salts thereof.
[0074] A further aspect of the invention includes pharmaceutical
compositions for treating conditions related to
glycosaminoglycan-associated molecular interactions, such as those
described supra. The therapeutic compounds in the methods of the
invention, as described hereinbefore, can be incorporated into a
pharmaceutical is composition in an amount effective to treat a
condition related to a glycosaminoglycan-associated molecular
interaction in a pharmaceutically acceptable carrier.
[0075] In another embodiment, the pharmaceutical compositions of
the invention include a therapeutic compound that has at least one
sulfate group covalently attached to a carrier molecule, or a
pharmaceutically acceptable salt thereof, in an amount sufficient
to treat a condition related to a glycosaminoglycan-associated
molecular interaction, and a pharmaceutically acceptable carrier.
The therapeutic compound can have the following formula:
QOSO.sub.3.sup.-X.sup.+].sub.n (III)
[0076] wherein Q is a carrier molecule; X.sup.+ is a cationic
group; and n is an integer selected such that the biodistribution
of the compound for an intended target site is not prevented while
maintaining activity of the compound.
[0077] The use of prodrugs which are converted in vivo to the
therapeutic compounds of the invention (see, e.g., R. B. Silverman,
1992, "The Organic Chemistry of Drug Design and Drug Action",
Academic Press, Chp. 8) are also to be considered within the scope
of the present invention. Such prodrugs can be used to alter the
biodistribution (e.g., to allow compounds which would not typically
cross the blood-brain barrier to cross the blood-brain barrier) or
the pharmacokinetics of the therapeutic compound. For example, an
anionic group, e.g., a sulfate or sulfonate, can be esterified,
e.g., with a methyl group or a phenyl group, to yield a sulfate or
sulfonate ester. When the sulfate or sulfonate ester is
administered to a subject, the ester is cleaved, enzymatically or
non-enzymatically, reductively or hydrolytically, to reveal the
anionic group. Such an ester can be cyclic, e.g., a cyclic sulfate
or sultone, or two or more anionic moieties may be esterified
through a linking group. Exemplary cyclic compounds include, for
example, 2-sulfobenzoic acid (LV), propane sultone (LVI), butane
sultone (LVII), 1,3-butanediol cyclic sulfate (LVIII,
.alpha.-chloro-.alpha.-hydroxy-o-toluenesulfonic acid sultone
(LIX), and 6-nitronaphth-[1,8-cd]-1,2,-oxathiole 2,2-dioxide (LX).
In an embodiment, the prodrug is a cyclic sulfate or sultone. An
anionic group can be esterified with moieties (e.g., acyloxymethyl
esters) which are cleaved to reveal an intermediate compound which
subsequently decomposes to yield the active compound. In another
embodiment, the prodrug is a reduced form of a sulfate or
sulfonate, e.g., a thiol, which is oxidized in vivo to the
therapeutic compound. Furthermore, an anionic moiety can be
esterified to a group which is actively transported in vivo, or
which is selectively taken up by target organs. The ester can be
selected to allow specific targeting of the therapeutic moieties to
particular organs, as described below for carrier moieties.
[0078] Carrier molecules useful in the therapeutic compounds
include carrier molecules previously described, e.g. carbohydrates,
polymers, peptides, peptide derivatives, aliphatic groups,
alicyclic groups, heterocyclic groups, aromatic groups or
combinations thereof. Suitable polymers include substituted and
unsubstituted vinyl, acryl, styrene and carbohydrate-derived
polymers and copolymers and salts thereof. Suitable carrier
molecules include a lower alkyl group, a heterocyclic group, a
disaccharide, a polymer or a peptide or peptide derivative.
[0079] Carrier molecules useful in the present invention may also
include moieties which allow the therapeutic compound to be
selectively delivered to a target organ or organs. For example, if
delivery of a therapeutic compound to the brain is desired, the
carrier molecule may include a moiety capable of targeting the
therapeutic compound to the brain, by either active or passive
transport (a "targeting moiety"). Illustratively, the carrier
molecule may include a redox moiety, as described in, for example,
U.S. Pat. Nos. 4,540,564 and 5,389,623, both to Bodor. These
patents disclose drugs linked to dihydropyridine moieties which can
enter the brain, where they are oxidized to a charged pyridinium
species which is trapped in the brain. Thus, drug accumulates in
the brain. Exemplary pyridine/dihdropyridine compounds of the
invention include sodium 1-(3-sulfopropyl)-1,4-dihydropyridine
(LXI), sodium 2-(nicotinylamido)-ethanesulfonate (LXII), and
1-(3-sulfopropyl)-pyridinium betaine (LXIII). Other carrier
moieties include compounds, such as amino acids or thyroxine, which
can be passively or actively transported in vivo. An illustrative
compound is phenylalanyltaurine (LXIX), in which a taurine molecule
is conjugated to a phenylalanine (a large neutral amino acid). Such
a carrier moiety can be metabolically removed in vivo, or can
remain intact as part of an active compound. Structural mimics of
amino adds (and other actively transported moieties) are also
useful in the invention (e.g.,
1-(aminomethyl)-1-(sulfomethyl)-cyclohexane (LXX)). Other exemplary
amino acid mimetics include p(sulfomethyl)phenylalanine (LXXII),
p-(1,3-disulfoprop-2-yl)phenylalanine (LXXIII), and
O-(1,3-disulfoprop-2-yl)tyrosine (LXXIV). Exemplary thyroxine
mimetics include compounds LXXV, LXVI, and LXXVII. Many targeting
moieties are known, and include, for example, asialoglycoproteins
(see, e.g. Wu, U.S. Pat. No. 5,166,320) and other ligands which are
transported into cells via receptor-mediated endocytosis (see below
for further examples of targeting moieties which may be covalently
or non-covalently bound to a carrier molecule). Furthermore, the
therapeutic compounds of the invention may bind to bacteria, viri,
infectious agents or cell membranes in the circulation and thus be
transported to the site of action.
[0080] The targeting and prodrug strategies described above can be
combined to produce a compound that can be transported as a prodrug
to a desired site of action and then unmasked to reveal an active
compound. For example, the dihydropyrine strategy of Bodor (see
supra) can be combined with a cyclic prodrug, as for example in the
compound
2-(1-methyl-1,4-dihydronicotinyl)amidomethyl-propanesultone
(LXXI).
[0081] In one embodiment, the therapeutic compound in the
pharmaceutical compositions is a sulfonated polymer, for example
poly(2-acrylamido-2-methyl-1-propanesulfonic acid);
poly(2-acrylamido-2-methyl-1-propanesulfonic
acid-co-acrylonitrile);
poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-styrene);
poly(vinylsulfonic acid); poly(sodium 4-styrenesulfonic acid); a
sulfonate derivative of poly(acrylic acid); a sulfonate derivative
of poly(methyl acrylate); a sulfonate derivative of poly(methyl
methacrylate); and a sulfonate derivative of poly(vinyl alcohol);
and pharmaceutically acceptable salts thereof.
[0082] The therapeutic compound can also have the structure:
##STR3##
[0083] in which Z is XR.sup.2 or R.sup.4, R.sup.1 and R.sup.2 are
each independently hydrogen, a substituted or unsubstituted
aliphatic group (preferably a branched or straight-chain aliphatic
moiety having from 1 to 24 carbon atoms in the chain; or an
unsubstituted or substituted cyclic aliphatic moiety having from 4
to 7 carbon atoms in the aliphatic ring; suitable aliphatic and
cyclic aliphatic groups are alkyl groups, more preferably lower
alkyl), an aryl group, a heterocyclic group, or a salt-forming
cation; R.sup.3 is hydrogen, lower alkyl, aryl, or a salt-forming
cation; X is, independently for each occurrence, O or S; R.sup.4 is
hydrogen, lower alkyl, aryl or amino; Y.sup.1 and Y.sup.2 are each
independently hydrogen, halogen (e.g., F, Cl, Br, or I), lower
alkyl, amino (including alkylamino, dialkylamino, arylamino,
diarylamino, and alkylarylamino), hydroxy, alkoxy, or aryloxy; and
n is an integer from 0 to 12 (more preferably 0 to 6, more
preferably 0 or 1). These compounds are described in U.S.
application Ser. No. 08/912,574, the contents of which are
incorporated herein by reference.
[0084] Suitable therapeutic compounds for use in the invention
include compounds in which both R.sup.1 and R.sup.2 are
pharmaceutically acceptable salt-forming cations. It will be
appreciated that the stoichiometry of an anionic compound to a
salt-forming counterion (if any) will vary depending on the charge
of the anionic portion of the compound (if any) and the charge of
the counterion. In a particularly suitable embodiment, R.sup.1,
R.sup.2 and R.sup.3 are each independently a sodium, potassium or
calcium cation. In certain embodiments in which at least one of
R.sup.1 and R.sup.2 is an aliphatic group, the aliphatic group has
between 1 and 10 carbons atoms in the straight or branched chain,
and is more preferably a lower alkyl group. In other embodiments in
which at least one of R.sup.1 and R.sup.2 is an aliphatic group,
the aliphatic group has between 10 and 24 carbons atoms in the
straight or branched chain. In certain embodiments, n is 0 or 1;
more preferably, n is 0. In certain embodiments of the therapeutic
compounds, Y.sup.1 and Y.sup.2 are each hydrogen.
[0085] In certain embodiments, the therapeutic compound of the
invention can have the structure: ##STR4##
[0086] in which R.sup.1, R.sup.2, R.sup.3, Y.sup.1, Y.sup.2, X and
n are as defined above. In other embodiments, the therapeutic
compound of the invention can have the structure: ##STR5##
[0087] in which R.sup.1, R.sup.2, R.sup.3, Y.sup.1, Y.sup.2, and X
are as defined above, R.sub.a and R.sub.b are each independently
hydrogen, alkyl, aryl, or heterocyclyl, or R.sub.a and R.sub.b,
taken together with the nitrogen atom to which they are attached,
form a cyclic moiety having from 3 to 8 atoms in the ring, and n is
an integer from 0 to 6. In certain embodiments, R.sub.a and R.sub.b
are each hydrogen. In certain embodiments, a compound of the
invention comprises an .alpha.-amino acid (or .alpha.-amino acid
ester), more preferably a L-.alpha.-amino acid or ester.
[0088] The Z, Q, R.sup.1, R.sup.2, R.sup.3, Y.sup.1, Y.sup.2 and X
groups are each independently selected such that the
biodistribution of the therapeutic compound for an intended target
site is not prevented while maintaining activity of the therapeutic
compound. For example, the number of anionic groups (and the
overall charge on the therapeutic compound) should not be so great
as to inhibit traversal of an anatomical barrier, such as a cell
membrane, or entry across a physiological barrier, such as the
blood-brain barrier, in situations where such properties are
desired. For example, it has been reported that esters of
phosphonoformate have biodistribution properties different from,
and in some cases superior to, the biodistribution properties of
phosphonoformate (see, e.g., U.S. Pat. Nos. 4,386,081 and 4,591583
to Helgstrand et al., and U.S. Pat. Nos. 5,194,654 and 5,463,092 to
Hostetler et al.). Thus, in certain embodiments, at least one of
R.sup.1 and R.sup.2 is an aliphatic group (more preferably an alkyl
group), in which the aliphatic group has between 10 and 24 carbons
atoms in the straight or branched chain. The number, length, and
degree of branching of the aliphatic chains can be selected to
provide a desired characteristic, e.g., lipophilicity. In other
embodiments, at least one of R.sup.1 and R.sup.2 is an aliphatic
group (more preferably an alkyl group), in which the aliphatic
group has between 1 and 10 carbons atoms in the straight or
branched chain. Again, the number, length, and degree of branching
of the aliphatic chains can be selected to provide a desired
characteristic, e.g., lipophilicity or ease of ester cleavage by
enzymes. In certain embodiments, a suitable aliphatic group is an
ethyl group.
[0089] In another embodiment, the therapeutic compound of the
invention can have the structure: ##STR6##
[0090] in which G represents hydrogen or one or more substituents
on the aryl ring (e.g., alkyl, aryl, halogen, amino, and the like)
and L is a substituted alkyl group (in certain embodiments,
preferably a lower alkyl), more preferably a hydroxy-substituted
alkyl or an alkyl substituted with a nucleoside base. In certain
embodiments, G is hydrogen or an electron-donating group. In
embodiments in which G is an electron-withdrawing group, G is
preferably an electron withdrawing group at the meta position. The
term "electron-withdrawing group" is known in the art, and, as used
herein, refers to a group which has a greater electron-withdrawing
than hydrogen. A variety of electron-withdrawing groups are known,
and include halogens (e.g., fluoro, chloro, bromo, and iodo
groups), nitro, cyano, and the like. Similarly, the term
"electron-donating group", as used herein, refers to a group which
is less electron-withdrawing than hydrogen. In embodiments in which
G is an electron donating group, G can be in the ortho, meta or
para position.
[0091] In certain embodiments, L is a moiety selected from the
group consisting of: ##STR7##
[0092] Table 1 lists data pertinent to the characterization of
these compounds using art-recognized techniques. TABLE-US-00001
TABLE 1 COMPOUND .sup.31P NMR .sup.13C NMR FAB-MS(-) IVa
-6.33(DMSO-d.sub.6) 60.97 CH.sub.2OH(d, J=6Hz) 245.2 66.76 CHOH(d,
J=7.8Hz) 121.65, 121.78, 121.99, 125.71, 129.48, 129.57, 126.43
Aromatic CH 134.38 Aniline C--N 150.39 Phenyl C--O(d, J=7Hz) 171.57
P--C.dbd.O(d, J=234Hz) IVb -6.41(DMSO-d.sub.6) 13.94 CH.sub.3 456
22.11, 24.40, 28.56, 28.72, 28.99, 29.00, 31.30, 33.43,
--(CH.sub.2).sub.10-- 65.03 CH.sub.2--OC(O) 66.60 CH.sub.2--OP(d,
J=5.6Hz) 67.71 CH2-OH(d, J=6Hz) 121.73, 121.10, 125.64, 126.57,
129.40, 129.95, Aromatic CH 134.04 Aniline C--N 150.31 Phenyl C--O
171.44 P--C.dbd.O(d, J=6.7Hz) 172.83 O--C.dbd.O IVc
-6.46(DMSO-d.sub.6) 13.94 CH.sub.3 471 22.11, 25.10, 28.68, 28.72,
28.85, 29.00, 30.76, 31.31, 32.10, --(CH.sub.2).sub.10-- 43.36
CH.sub.2--S 68.43 CH.sub.2--OH 68.43 CH--OH(d, J=6.3Hz) 68.76
P--O--CH.sub.2-9d, J=5.8Hz) 121.75, 122.03, 125.62, 126.37, 129.30,
129.53, Aromatic CH 134.23 Aniline C--N 150.37 Phenyl C--O(d,
J=6.7Hz) 171.47 P--C.dbd.O(d, J=234.0Hz) 198.47 S--C.dbd.O IVd
-6.61(DMSO-d.sub.6) .sub.13.94 CH.sub.3 416 22.06, 25.14, 28.24,
28.35, 31.09, 32.14 --CH.sub.2).sub.6- 43.40 CH.sub.2--S 68.50
P--O--CH.sub.2--(d, J=5.8Hz) 68.77 CH--OH(d, 6.4Hz) 121.78, 122.59,
125.69, 127.06, 129.43, 129.59 Aromatic CH 133.39 Aniline C--N
150.38 Phenyl C--O(d, J=6.7Hz) 171.47 P--C.dbd.O(d, J=234.4Hz)
198.54 S--C.dbd.O IVe -5.76(D.sub.2O) N/A N/A IVf
-7.00(DMSO-d.sub.6) N/A N/A IVg -6.60(DMSO-D6) 70.84 CH2-OH 321
72.17 CH--OH 121.68, 121.79, 121.85, 125.71 127.10, 127.92, 129.36,
129.50, 129.59 Aromatic CH 134.51 Aniline C--N 142.34 Aromatic
C--CH 150.37 Phenyl C--O(d, J=6.2Hz) 171.59 P--C.dbd.O(d,
J=232.6Hz)
[0093] It will be noted that the structure of some of the
therapeutic compounds of this invention includes asymmetric carbon
atoms. It is to be understood accordingly that the isomers (e.g.,
enantiomers and diastereomers) arising from such asymmetry are
included within the scope of this invention. Such isomers can be
obtained in substantially pure form by classical separation
techniques and by sterically controlled synthesis. For the purposes
of this application, unless expressly noted to the contrary, a
therapeutic compound shall be construed to include both the R or S
stereoisomers at each chiral center.
[0094] In certain embodiments, an therapeutic compound of the
invention comprises a cation (i.e., in certain embodiments, at
least one of R.sup.1, R.sup.2 or R.sup.3 is a cation). If the
cationic group is hydrogen, H.sup.+, then the therapeutic compound
is considered an acid, e.g., phosphonoformic acid. If hydrogen is
replaced by a metal ion or its equivalent, the therapeutic compound
is a salt of the acid. Pharmaceutically acceptable salts of the
therapeutic compound are within the scope of the invention. For
example, at least one of R.sup.1, R.sup.2 or R.sup.3 can be a
pharmaceutically acceptable alkali metal (e.g., Li, Na, or K),
ammonium cation, alkaline earth cation (e.g., Ca.sup.2+, Ba.sup.2+,
Mg.sup.2+), higher valency cation, or polycationic counter ion
(e.g., a polyammonium cation). (See, e.g., Berge et al. (1977)
"Pharmaceutical Salts", J. Pharm. Sci. 66:1-19). It will be
appreciated that the stoichiometry of an anionic compound to a
salt-forming counterion (if any) will vary depending on the charge
of the anionic portion of the compound (if any) and the charge of
the counterion. Preferred pharmaceutically acceptable salts include
a sodium, potassium or calcium salt, but other salts are also
contemplated within their is pharmaceutically acceptable range.
[0095] The term "pharmaceutically acceptable esters" refers to the
relatively non-toxic, esterified products of the therapeutic
compounds of the present invention. These esters can be prepared in
situ during the final isolation and purification of the therapeutic
compounds or by separately reacting the purified therapeutic
compound in its free acid form or hydroxyl with a suitable
esterifying agent; either of which are methods known to those
skilled in the art Carboxylic acids and phosphonic acids can be
converted into esters according to methods well known to one of
ordinary skill in the art, e.g., via treatment with an alcohol in
the presence of a catalyst. A preferred ester group (e.g., when
R.sup.3 is lower alkyl) is an ethyl ester group.
[0096] The term "alkyl" refers to the saturated aliphatic groups,
induding straight-chain alkyl groups, branched-chain alkyl groups,
cycloalkyl (alicyclic) groups, alkyl substituted cydoalkyl groups,
and cycloalkyl substituted alkyl groups. In preferred embodiments,
a straight chain or branched chain alkyl has 30 or fewer carbon
atoms in its backbone (e.g., C.sub.1-C.sub.30 for straight chain,
C.sub.3-C.sub.30 for branched chain), and more preferably 20 or
fewer. Likewise, preferred cycloalkls have from 4-10 carbon atoms
in their ring structure, and more preferably have 4-7 carbon atoms
in the ring structure. The term "lower alkyl" refers to alkyl
groups having from 1 to 6 carbons in the chain, and to cycloalkls
having from 3 to 6 carbons in the ring structure.
[0097] Moreover, the term "alkyl" (including "lower alkyl") as used
throughout the specification and claims is intended to include both
"unsubstituted alkyls" and "substituted alkyls", the latter of
which refers to alkyl moieties having substituents replacing a
hydrogen on one or more carbons of the hydrocarbon backbone. Such
substituents can include, for example, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkyl amino, dialkylamino,
arylamino, diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfate, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl,
cyano, azido, heterocyclyl, aralkyl, or an aromatic or
heteroaromatic moiety. It will be understood by those skilled in
the art that the moieties substituted on the hydrocarbon chain can
themselves be substituted, if appropriate. Cycloalkyls can be
further substituted, e.g., with the substituents described above.
An "aralkyl" moiety is an alkyl substituted with an aryl (e.g.,
phenylmethyl(benzyl)).
[0098] The term "alkoxy", as used herein, refers to a moiety having
the structure --O-alkyl, in which the alkyl moiety is described
above.
[0099] The term "aryl" as used herein includes 5- and 6-membered
single-ring aromatic groups that may include from zero to four
heteroatoms, for example, unsubstituted or substituted benzene,
pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole,
pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the
like. Aryl groups also include polycyclic fused aromatic groups
such as naphthyl, quinolyl, indolyl, and the like. The aromatic
ring can be substituted at one or more ring positions with such
substituents, e.g., as described above for alkyl groups. Preferred
aryl groups include unsubstituted and substituted phenyl
groups.
[0100] The term "aryloxy", as used herein, refers to a group having
the structure --O-aryl, in which the aryl moiety is as defined
above.
[0101] The term "amino," as used herein, refers to an unsubstituted
or substituted moiety of the formula --NR.sub.aR.sub.b, in which
R.sub.a and R.sub.b are each independently hydrogen, alkyl, aryl,
or heterocyclyl, or R.sub.a and R.sub.b, taken together with the
nitrogen atom to which they are attached, form a cyclic moiety
having from 3 to 8 atoms in the ring. Thus, the term "amino" is
intended to include cyclic amino moieties such as piperidinyl or
pyrrolidinyl groups, unless otherwise stated. An "amino-substituted
amino group" refers to an amino group in which at least one of
R.sub.a and R.sub.b, is further substituted with an amino
group.
[0102] In another embodiment, R.sup.1 or R.sup.2 can be (for at
least one occurrence) a long-chain aliphatic moiety. The term
"long-chain aliphatic moiety" as used herein, refers to a moiety
having a straight or branched chain aliphatic moiety (e.g., an
alkyl or alkenyl moiety) having from 10 to 24 carbons in the
aliphatic chain, e.g., the long-chain aliphatic moiety is an
aliphatic chain of a fatty acid (preferably a naturally-occurring
fatty acid). Representative long-chain aliphatic moieties include
the aliphatic chains of stearic acid, oleic acid, linolenic add,
and the like.
[0103] In certain embodiments, the therapeutic compound of the
invention can have the structure: ##STR8##
[0104] in which R.sup.1 and R.sup.2 are each independently
hydrogen, an aliphatic group (preferably a branched or
straight-chain aliphatic moiety having from 1 to 24 carbon atoms,
more preferably 10-24 carbon atoms, in the chain; or an
unsubstituted or substituted cyciic aliphatic moiety having from 4
to 7 carbon atoms in the aliphatic ring), an aryl group, a
heterocyclic group, or a salt-forming cation; R.sup.3 is hydrogen,
lower alkyl, aryl, or a salt-forming cation; Y.sup.1 and Y.sup.2
are each independently hydrogen, halogen (e.g., F, Cl, Br, or I),
lower alkyl, hydroxy, alkoxy, or aryloxy; and n is an integer from
0 to 12. Suitable therapeutic compounds for use in the invention
include compounds in which both R.sup.1 and R.sup.2 are
pharmaceutically acceptable salt-forming cations. In a particularly
suitable embodiment, R.sup.1, R.sup.2 and R.sup.3 are each
independently a sodium, potassium or calcium cation, and n is 0. In
certain embodiments of the therapeutic compounds, Y.sup.1 and
Y.sup.2 are each hydrogen. Suitable therapeutic compounds include
salts of phosphonoformate. Trisodium phosphonoformate (foscarnet
sodium or Foscavir.RTM.) is commercially available (e.g., from
Astra), and its clinical pharmacology has been investigated (see,
e.g., "Physician's Desk Reference", 51st Ed., pp. 541-545
(1997)).
[0105] In another embodiment, the therapeutic compound used in the
invention can be an aminophosphonate, a biphosphonate, a
phosphonocarboxylate derivative, a phosphonate derivative, or a
phosphono carbohydrate. For example, the therapeutic compound can
be one of the compounds described in Appendix A submitted
herewith.
[0106] Suitable therapeutic compounds for inclusion in a
pharmaceutical composition for treating
glycosaminoglycan-associated molecular interactions include
1,3-propanedisulfonic acid, 3-amino-1-propanesulfonic acid,
3-dimethylamino-1-propanesulfonic acid sodium salt,
2-(3-sulfopropyl)-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole, sodium
salt,
3-[2-(6-methoxy-1,2,3,4-tetrahydroisoquinolinyl)]-1-propanesulfonic
acid, 3-(2-hydroxyethyl)amino-1-propanesulfonic add,
3-(3-hydroxy-1-propyl)amino-1-propanesulfonic acid,
(-)3-[(R)-2-hydroxy-1-propyl]amino-1-propanesulfonic acid,
3-(4-hydroxy-1-butyl)amino-1-propanesulfonic acid,
3-(5-hydroxy-1-pentyl)amino-1-propanesulfonic acid,
3-(6-hydroxy-1-hexyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-(2-hydroxyethyl)amino-1-propanesulfonic acid,
3-hexylamino-1-propanesulfonic acid,
3-undecylamino-1-propanesulfonic acid, and
3-octadecylamino-1-propanesulfonic acid, and pharmaceutically
acceptable salts or esters thereof.
[0107] In the methods of the invention, a condition related to a
glycosaminoglycan-associated molecular interaction in a subject is
treated by administering a therapeutic compound of the invention to
the subject. The term "subject" is intended to include living
organisms in which conditions related to
glycosaminoglycan-associated molecular interactions can occur. The
term subject is also intended to include those living organisms
which are afflicted by infectious agents which secrete components
which interfere with a host cells via glycosaminoglycans. Examples
of subjects include humans, monkeys, cows, sheep, goats, dogs,
cats, mice, rats, and transgenic species thereof. Administration of
the 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 treat a condition related to a
glycosaminoglycan-associated molecular interaction in the subject.
An effective amount of the therapeutic compound necessary to
achieve a therapeutic effect may vary according to factors such as
the age, sex, and weight of the subject, and the ability of the
therapeutic compound to treat the foreign agents in the subject.
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. A non-limiting example of
an effective dose range for a therapeutic compound of the invention
(e.g., poly(vinylsulfonate sodium salt)) is between 5 and 500 mg/kg
of body weight/per day. In an aqueous composition, suitable
concentrations for the active compound (i.e., the therapeutic
compound that can treat the disease) are between 5 and 500 mM,
between 10 and 100 mM, and between 20 and 50 mM.
[0108] The therapeutic compounds of the invention may be
administered orally. Alternatively, the active compound may be
administered by other suitable routes such subcutaneous,
intravenous, intraperitoneal, etc. administration (e.g. by
injection). Depending on the route of administration, the active
compound may be coated in a material to protect the compound from
the action of acids and other natural conditions which may
inactivate the compound.
[0109] The compounds of the invention can be formulated to ensure
proper distribution in vivo. For example, the blood-brain barrier
(BBB) exdudes many highly hydrophilic compounds. To ensure that the
therapeutic compounds of the invention cross the BBB, they can be
formulated, for example, in liposomes. For methods of manufacturing
liposomes, see, e.g., U.S. Pat. Nos. 4,522,811; 5,374,548; and
5,399,331. The liposomes may comprise one or more moieties which
are selectively transported into specific cells or organs
("targeting moieties"), thus providing targeted drug delivery (see,
e.g., V. V. Ranade (1989) J. Clin. Pharmacol. 29:685). Exemplary
targeting moieties include folate or biotin (see, e.g., U.S. Pat.
No. 5,416,016 to Low et al.); mannosides (Umezawa et al., (1988)
Biochem. Biophys. Res. Commun. 153:1038); antibodies (P. G. Bloeman
et al. (1995) FEBS Lett. 357:140; M. Owais et al. (1995)
Antimicrob. Agents Chemother. 39:180); surfactant protein A
receptor (Briscoe et al. (1995) Am. J. Physiol. 1233:134); gp120
(Schreier et al. (1994) J. Biol. Chem. 269:9090); see also K.
Keinanen; M. L. Laukkanen (1994) FEBS Lett. 346:123; J. J. Killion;
I. J. Fidler (1994) Immunomethods 4:273. In an embodiment, the
therapeutic compounds of the invention are formulated in liposomes;
in a more preferred embodiment, the liposomes include a targeting
moiety.
[0110] Delivery and in vivo distribution can also be affected by
alteration of an anionic group of compounds of the invention. For
example, anionic groups such as carboxylate or tetrazole can be
employed instead of, or in addition to, sulfate or sulfonate
moieties, to provide compounds with desirable pharmocokinetic,
pharmacodynamic, biodistributive, or other properties. Exemplary
tetrazole-substituted compounds include
3-(1H-tetrazol-5-yl)-9H-thioxanthen-9-one 10,10-dioxide (LXIV),
5,5-dithiobis(1-phenyltetrazole) (LXV), 1H-tetrazole (LXVI),
5-phenyl-1H-tetrazole (LXVII), and 5-(2-aminoethanoic
acid)-1H-tetrazole (LXVIII), and the like; and their
pharmaceutically acceptable salts. Exemplary
carboxylate-substituted compounds include dicarboxylic acids such
as adipic add, azelaic acid, 3,3-dimethylglutaric acid, suberic
acid, succinic acid, and the like, and their pharmaceutically
acceptable salts.
[0111] To administer the therapeutic compound by other than
parenteral administration, it may be necessary to coat the compound
with, or co-administer the compound with, a material to prevent its
inactivation. For example, the therapeutic compound 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 (Strejan et al., (1984) J. Neuroimmunol. 7:27).
[0112] The therapeutic compound may also be administered
parenterally, intraperitoneally, intraspinally, or intracerebrally.
Dispersions 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.
[0113] 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.
The carrier 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, it will be
preferable to include isotonic agents, 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.
[0114] Sterile injectable solutions can be prepared by
incorporating the therapeutic compound 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 compound into a sterile carrier 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 preferred methods
of preparation are vacuum drying and freeze-drying which yields a
powder of the active ingredient (i.e., the therapeutic compound)
plus any additional desired ingredient from a previously
sterile-filtered solution thereof.
[0115] The therapeutic compound can be orally administered, for
example, with an inert diluent or an assimilable edible carrier.
The therapeutic compound and other ingredients may also be enclosed
in a hard or soft shell gelatin capsule, compressed into tablets,
or incorporated directly into the subject's diet. For oral
therapeutic administration, the therapeutic compound may be
incorporated with excipients and used in the form of ingestible
tablets, buccal tablets, troches, capsules, elixirs, suspensions,
syrups, wafers, and the like. The percentage of the therapeutic
compound in the compositions and preparations may, of course, be
varied. The amount of the therapeutic compound in such
therapeutically useful compositions is such that a suitable dosage
will be obtained.
[0116] It is especially advantageous to formulate parenteral
compositions in dosage unit form for ease of administration and
uniformity of dosage. Dosage unit form as used herein refers to
physically discrete units suited as unitary dosages for the
subjects to be treated; each unit containing a predetermined
quantity of therapeutic compound calculated to produce the desired
therapeutic effect in association with the required pharmaceutical
carrier. The specification for the dosage unit forms of the
invention are dictated by and directly dependent on (a) the unique
characteristics of the therapeutic compound and the particular
therapeutic effect to be achieved, and (b) the limitations inherent
in the art of compounding such a therapeutic compound for the
treatment of a condition related to a glycosaminoglycan-associated
molecular interaction in a subject.
[0117] Active compounds are administered at a therapeutically
effective dosage sufficient to treat a condition related to a
glycosaminoglycan-associated molecular interaction in a subject. A
"therapeutically effective dosage" preferably reduces the amount of
symptoms of the condition in the infected subject by at least about
20%, more preferably by at least about 40%, even more preferably by
at least about 60%, and still more preferably by at least about 80%
relative to untreated subjects. For example, the ability of a
compound to reduce an infectious agent can be evaluated in an
animal model system that may be predictive of efficacy in treating
diseases associated with the infectious agent in humans.
[0118] The invention is further illustrated by the following
Exemplification, which should not be construed as further limiting
the subject invention. The contents of all references, issued
patents, and published patent applications cited throughout this
application including the background are hereby incorporated by
reference.
EXEMPLIFICATION
Bacteria:
Streptococcus pyogens:
[0119] S. pyogenes can cause acute rheumatic fever streptococcus
and acute post-streptococcal glomerulonephritis. Studies have
identified the protein responsible for stabilizing the bacteria on
the basal laminae of cardiac muscle as well as on kidney
tissues.
[0120] The mechanism for such a bacterial virulence is unknown. One
hypothesis is the direct binding of bacterial adhesions and
exotoxins to the cardiac muscle and kidney_Bergey & Stenson
identified two streptococcal proteins (9.15 Kda) that are capable
of binding to basement membranes of cardiac muscle and renal
tissues; the binding was completely inhibited by heparin and other
GAGs. Binding of specific S. pyogenes protein can be measured on
cardiac muscle section and kidney section. Briefly, S. pyogenes
protein preparation are incubated on cardiac muscle and kidney
preparation. Binding of protein is visualized by indirect
immunofluorescence using an antibody against the bacteria protein.
Ability of a compound to interfere in such a binding can be
determined in ligand inhibition studies. Binding is measured by
comparing the amount of protein binding (as determined by Image
Analysis) in presence or absence of the compound. Measurement can
also be determined using 3H-labeled streptococcal protein.
[0121] Fluorescence: Cryostat-cut section of heart tissue are
pre-incubated with bacterial antigen preparation. Amount of peptide
binding to tissue is determined by indirect immunofluorescence
using a Rabbit anti-S. pyogenes serum. Ability of a compound to
inhibit the binding of the streptococcal protein to the tissue is
evaluated by comparing the amount of bacterial antigen present of
tissue section in presence or absence of the inhibitor.
[0122] Direct Binding Assay: Radiolabefled streptococcal components
are tested for direct binding activity to mammalian tissue
component as previously described (M W Stinson & E J Bergey,
1982). Dried cardiac muscle fragments are rehydrated with PBS and
1% bovine serum albumin. Moist heart material is incubated with
radiolabelled bacterial components. Bound radioactivity is
determined by liquid scintillation spectrometry.
[0123] Determination of a compound's ability to inhibit this
binding is done with various concentrations of the compound added
to the streptococcal preparation prior its incubation with the
cardiac muscle preparation.
[0124] S. aureus, Pseudomonas aeruglnosa, Legionella
pneumophila
[0125] S. aureus and P. aeruginosa are well-known to cause major
pulmonary infection in patients with cystic fibrosis. Legionella
pneumophila is known to cause Legionnaire's disease in susceptible
individuals. These bacteria need to adhere to mucus membrane to in
order to multiply and cause infection.
[0126] The ability of specific compounds to inhibit S. Aureus, P.
Aeruginosa and L. pneumophila adherence to mucosal membrane can be
determined in vitro using murine trachea culture. The number of
bacteria adhering to the preparation can be determined by comparing
the number of bacteria remaining in supernatant after incubation
with trachea preparation. Briefly, trachea preparation are
incubated with a bacterial suspension in presence or absence of a
compound. 30 minutes later the amount of bacteria remaining the
supernatant (i.e., non-adhering) determined by serial dilution.
[0127] The ability of bacteria to infect cells an also be
determined in vitro. Macrophages are incubated with bacteria and
the phagocytic rate is determined 30 minutes later.
[0128] In vivo: Intratracheal infection with P. aeruginosa or S.
aureus with or without treatment with compound. Intratracheal
infection with P. aeruginosa, S. aureus and Legionella have been
shown to cause acute pulmonary infection in mice. The ability of a
compound to inhibit such an infection can be determined by
evaluating the bacterial load present in the lung of infected mice
undergoing a treatment with specific compounds. These compounds can
be administered IV, PO, or under aerosol.
[0129] Viral Infections:
[0130] The infectious process of viruses of the herpesviridae
family have been extensively studied. It has been established that
the initial interaction of several herpes viruses with the cell
surface is mediated by glycosaminoglycans found on the
proteoglycans in the cell plasma membrane. These GAGs are similar
to heparin. Amongst the different herpes viruses found to interact
with cell surface GAGs, interesting ones are Cytomegalovirus (CMV)
and Herpes simplex (HSV-1 and HSV-2). The ability of compounds to
interfere in the infectious process of these viruses is determined
as follows:
[0131] In vitro: Hela cells are infected with CMV or HSV-1 in
presence or absence of compounds. Ability of CMV to infect cells is
determined by evaluating virus load 24-72 hours later by: [0132] %
viral antigen expression (IF) [0133] specific viral antigen (mRNA
level) [0134] Virus particle titration [0135] cytopathic effect
[0136] The ability of a compound to interfere in the infectious
process can be determined by evaluating (by the different
techniques mentioned above) the amount of virus found in the
culture in presence or absence of an inhibitor.
EQUIVALENTS
[0137] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures described herein. Such
equivalents are considered to be within the scope of this invention
and are covered by the following claims.
APPENDIX A
[0138] TABLE-US-00002 AMINOPHOSPHONATES Name Structure
3-[2-(1,2,3,4-Tetrahydroisoquinolinyl)]-1-pro- panephosphonic acid,
disodium salt ##STR9## 3-Aminopropylphosphonic acid
NH.sub.2CH.sub.2CH.sub.2CH.sub.2PO.sub.3H.sub.2
(S)-2-Amino-2-methyl-4-phos- phonobutanoic acid ##STR10##
D-(-)-2-Amino-4-phosphonobutanoic acid ##STR11##
L-(+)-2-Amino-4-phosphonobutanoic acid ##STR12##
3-Aminopropyl(methyl)phosphinic acid, hydrochloride ##STR13##
(R)-(-)-3-(2-Carboxypiperazin-4-yl)-pro- pyl-1-phosphonic
acid(D-CPP) ##STR14## (R,E)-4-(3-Phosphonoprop-2-en-
yl)piperazine-2-carboxylic acid ##STR15##
trans-L-4-Phosphonomethylproline, trisodium salt ##STR16##
cis-L-4-Phosphonomethylproline, trisodium salt ##STR17##
4-Amino-1-butylphosphonic acid, disodium salt ##STR18##
1-(3-Phosphonopropyl)-benzimidazole, disodium salt ##STR19##
3-Dimethylamino-1-propylphosphonic acid, disodium salt ##STR20##
3-Amino-butylphosphonic acid, disodium salt ##STR21##
3-Amino-pentylphosphonic acid, disodium salt ##STR22##
3-Amino-hexylphosphonic acid, disodium salt ##STR23##
3-Amino-heptylphosphonic acid, disodium salt ##STR24##
3-Amino-octylphophonic acid, disodium salt ##STR25##
3-Amino-4-methyl-pentylphosphonic acid, disodium salt ##STR26##
3-Amino-3-methyl-butylphosphonic acid, disodium salt ##STR27##
3-Amino-3-phenyl-propylphosphonic acid, disodium salt ##STR28##
3-Amino-4-phenyl-butylphosphonic acid, disodium salt ##STR29##
3-Amino-4-phenyl-pentylphosphonic acid, disodium salt ##STR30##
3-Amino-3-phenyl-butylphosphonic acid, disodium salt ##STR31##
2-Amino-2-(2-phosphonoethyl)-1,3,4-tri- hydronaphthalene, disodium
salt ##STR32## 1-Amino-1-(2-phosphonoethyl)-cyclo- hexane, disodium
salt ##STR33## 2-(2-Amino-4-phos- phonobutoxy)tetrahydropyran
##STR34## 3-Amino-4-hydroxy-butylphosphonic acid, disodium salt
##STR35## Diethyl 2-pyrrolidinylphosphonate ##STR36##
2-Pyrrolidinylphosphonic acid, disodium salt ##STR37##
1,1-Dioxo-2-(3-phosphonopropyl)-iso- thiazoline, disodium salt
##STR38## 2-Deoxy-2-phosphonoacetylamino-D-glucose ##STR39##
3-Hydroxy-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt
##STR40## 3-Hydroxy-3-(3-pyridyl)propenyl-2-phos- phonic acid,
disodium salt ##STR41## 3-Hydroxy-3-(4-pyridyl)propenyl-2-phos-
phonic acid, disodium salt ##STR42##
3-Amino-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt
##STR43## 3-Amino-3-(3-pyridyl)propenyl-2-phos- phonic acid,
disodium salt ##STR44## 3-Amino-3-(4-pyridyl)propenyl-2-phos-
phonic acid, disodium salt ##STR45##
1,4-Diamino-1-(3-pyridyl)butyl-2-phos- phonic acid, disodium salt
##STR46## 1,4-Diamino-4-methyl-1-(3-py- ridyl)pentyl-2-phosphonic
acid, disodium salt ##STR47## 1,4-Diamino-4-methyl-1-(2-py-
ridyl)pentyl-2-phosphonic acid, disodium salt ##STR48##
1,4-Diamino-4-methyl-1-(4-py- ridyl)pentyl-2-phosphonic acid,
disodium salt ##STR49## 3-(2-Amino-4,5,7,8-tetrahydro-6H-thia-
zolo[4,5-d]azepin-6-yl)propyl- phosphonic acid, disodium salt
##STR50## N-Phosphonomethylglycine ##STR51##
N-Phosphonomethylglycine, trisodium salt ##STR52##
(2R,4S)-4-Phosphonomethylpipecolinic acid, trisodium salt ##STR53##
(2R,4S)-4-Phosphonomethyl- pipecolinamide, disodium salt ##STR54##
N-Phosphonomethylglycine (Aldrich, see NC-1769) ##STR55##
N-Phosphonomethylglycine, trisodium salt (see NC1770, prepared from
NC1781) ##STR56## 3-[6-Methoxy-2-(1,2,3,4-tetrahydro-
isoquinolinyl)]propylphosphonic acid, disodium salt ##STR57##
3-[8-Methoxy-2-(1,2,3,4-tetrahydro- isoquinolinyl)]propylphosphonic
acid, disodium salt ##STR58##
3-[2-(3-Methoxycarbonyl-1,2,3,4-tetra- hydroisoquinolinyl)]-propyl-
phosphonic acid disodium salt ##STR59##
2-(3-Phosphonopropyl)-1,2,3,4-tetra- hydro-9H-pyrido[3,4-b]indole,
disodium salt ##STR60##
[0139] TABLE-US-00003 Bisphosphonates Name Structure Pamidronic
acid(3-Aminopropyl-1-hydroxy- propane-1,1-bisphosphonic acid)
##STR61## 3-Amino-1-hydroxypropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR62## 1-Amino-3-sulfopropane-1,1-bis-
phosphonic acid ##STR63## 1-Amino-3-sulfopropane-1,1-bis-
phosphonic acid, pentasodium salt ##STR64##
1,3-Diaminopropane-1,1-bisphosphonic acid, tetrasodium salt
##STR65## 1-Amino-3-dimethylaminopropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR66##
3-Dimethylamino-1-hydroxypropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR67## 1-Hydroxy-3-(methylphenylamino)-pro-
pane-1,1-bisphosphonic acid, tetrasodium salt ##STR68##
1-Amino-3-(methylphenylamino)propane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR69## Ibandronic acid, tetrasodium salt
(1-Hydroxy-3-(methylpentylamino)-pro- pane-1,1-bisphosphonic acid,
tetrasodium salt) ##STR70##
1-Amino-3-(methylpentylamino)propane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR71##
1-Amino-3-(1-benzimidazolyl)propane-1,1-bis- phosphonic acid
##STR72## 1-Amino-3-(1-benzimidazolyl)propane-1,1-bis- phosphonic
acid, tetrasodium salt ##STR73## 3-Aminopropane-1,1-bisphosphonic
acid, tetrasodium salt ##STR74##
(dl)-3-Aminobutane-1,1-bisphosphonic acid, tetrasodium salt
##STR75## (dl)-3-Aminopentane-1,1-bisphosphonic acid, tetrasodium
salt ##STR76## (dl)-3-Aminohexane-1,1-bisphosphonic acid,
tetrasodium salt ##STR77## (dl)-3-Aminoheptane-1,1-bisphosphonic
acid, tetrasodium salt ##STR78##
(dl)-3-Aminooctane-1,1-bisphosphonic acid, tetrasodium salt
##STR79## (dl)-3-Amino-4-methylpentane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR80## (dl)-3-Amino-3-methylbutane-1,1-bis-
phosphonic acid, tetrasodium salt ##STR81##
(dl)-3-Amino-3-phenylpropane-1,1-bis- phosphonic acid, tetrasodium
salt ##STR82## (dl)-3-Amino-4-phenylbutane-1,1-bis- phosphonic
acid, tetrasodium salt ##STR83##
(dl)-3-Amino-4-phenylpentane-1,1-bis- phosphonic acid, tetrasodium
salt ##STR84## (dl)-3-Amino-3-phenylbutane-1,1-bis- phosphonic
acid, tetrasodium salt ##STR85## (dl)-2-(2-Amino-1,2,3,4-tetra-
hydronaphthalenyl)ethane-1,1-bis- phosphonic acid, tetrasodium salt
##STR86## 2-(1-Aminocyclohexyl)ethane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR87##
2-(2-Amino-4,4-bisphosphonobutoxy)-tetra- hydropyran, tetrasodium
salt ##STR88## (dl)-3-Amino-4-hydroxybutane-1,1-bis- phosphonic
acid, tetrasodium salt ##STR89##
(S)-Hydroxy(2-pyrrolidinyl)methane- bisphosphonic acid tetrasodium
salt ##STR90## Hydroxy[(2S,4R)-4-hydroxy-2-pyr-
rolidinyl]methanebisphosphonic acid tetrasodium salt ##STR91##
2-Amino-1-hydroxyethane-1,1-bis- phosphonic acid, tetrasodium salt
##STR92## 1,2-Diaminoethane-1,1-bisphonponic acid, tetrasodium salt
##STR93## 4-Amino-1-hydroxybutane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR94## 1,4-Diaminobutane-1,1-bisphosphonic
acid, tetrasodium salt ##STR95## 5-Amino-1-hydroxypentane-1,1-bis-
phosphonic acid, tetrasodium salt ##STR96##
1,5-Diaminopentane-1,1-bisphosphonic acid, tetrasodium salt
##STR97## (S)-2-Amino-1-hydroxypropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR98## (S)-2-Amino-1-hydroxybutane-1,1-bis-
phosphonic acid, tetrasodium salt ##STR99##
(S)-2-Amino-1-hydroxy-3-methylbutane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR100##
(S)-2-Amino-1-hydroxy-3-phenylpropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR101##
(S)-2-Amino-1,3-dihydroxypropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR102##
(S)-2,3-Diamino-1-hydroxypropane-1,1-bis- phosphonic acid,
tetrasodium salt ##STR103## (dl)-3-Amino-1-hydroxy-3-phenyl-
propane-1,1-bisphosphonic acid, tetrasodium salt ##STR104##
(S)-3-Amino-2-(4-chlorophenyl)-1-hy- droxypropane-1,1-bisphosphonic
acid, tetrasodium salt ##STR105##
(S)-2-Amino-3-(4-aminophenyl)-1-hy- droxypropane-1,1-bisphosphonic
acid, tetrasodium salt ##STR106##
[0140] TABLE-US-00004 Phosphonocarboxylate Derivatives Name
Structure Phosphonoacetic acid(fosfonet) ##STR107## Phosphonoformic
acid, trisodium salt ##STR108## Diethylphosphonoacetic acid
##STR109## 2-Carboxyethylphosphonic acid
HO.sub.2CCH.sub.2CH.sub.2PO.sub.3H.sub.2
(dl)-2-Amino-3-phosphonopropanoic acid ##STR110##
(dl)-2-Amino-5-phosphonopentanoic acid ##STR111## Phosphonoacetic
acid(See NC-769) HO.sub.2CCH.sub.2PO.sub.3H.sub.2
(S)-2-Amino-2-methyl-4-phos- phonobutanoic acid ##STR112##
D-(-)-2-Amino-4-phosphonobutanoic acid ##STR113##
L-(+)-2-Amino-4-phosphonobutanoic acid ##STR114##
D-(-)-2-Amino-7-phosphonoheptanoic acid ##STR115##
L-(+)-2-Amino-7-phosphonoheptanoic acid ##STR116##
D-(-)-2-Amino-6-phosphonohexanoic acid ##STR117##
L-(+)-2-Amino-6-phosphonohexanoic acid ##STR118##
D-(-)-2-Amino-4-phosphonopentanoic acid ##STR119##
L-(+)-2-Amino-4-phosphonopentanoic acid ##STR120##
D-(-)-2-Amino-3-phosphonopropanoic acid ##STR121##
L-(+)-2-Amino-3-phosphonopropanoic acid ##STR122##
(R)-(-)-3-(2-Carboxypiperazin-4-yl)-pro- pyl-1-phosphonic
acid(D-CPP) ##STR123## L-4-[Difluoro(phosphono)methyl)]-phenyl-
alanine ##STR124## (R,E)-4-(3-Phosphonoprop-2-en-
yl)piperazine-2-carboxylic acid ##STR125##
trans-L-4-Phosphonomethylproline, trisodium salt ##STR126##
cis-L-4-Phosphonomethylproline, trisodium salt ##STR127##
N,N-Diethylphosphonoacetamide, disodium salt ##STR128##
N-Cyclohexylphosphonoacetamide, disodium salt ##STR129##
Phosphonoacetic hydrazide, disodium salt ##STR130##
N-Hydroxyphosphonoacetamide, disodium salt ##STR131##
N-Phosphonoacetyl-L-alanine, trisodium salt ##STR132##
N-Phosphonoacetyl-L-glycine, trisodium salt ##STR133##
N-(Phosphonoactyl)-L-asparagine-L-glycine, tetrasodium salt
##STR134## N-Phosphonomethylglycine ##STR135##
N-Phosphonomethylglycine, trisodium salt ##STR136##
2-Phosphonomethylglutaric acid, tetrasodium salt ##STR137##
2-Phosphonomethylsuccinic acid, tetrasodium salt ##STR138##
(2R,4S)-4-Phosphonomethylpipecolinic acid, trisodium salt
##STR139## (2R,4S)-4-Phosphonomethyl- pipecolinamide, disodium salt
##STR140## N-Phosphonomethylglycine (Aldrich, see NC-1769)
##STR141## N-Phosphonomethylglycine, trisodium salt (see NC1770,
prepared from NC1781) ##STR142##
3-[2-(3-Methoxycarbonyl-1,2,3,4-tetra- hydroisoquinolinyl)]-pro-
pylphosphonic acid disodium salt ##STR143##
[0141] TABLE-US-00005 Phosphonate derivative Name Structure
3-[2-(1,2,3,4-Tetrahydroisoquinolinyl)]-1-pro- panephosphonic acid,
disodium salt ##STR144## Propylphosphonic acid
CH.sub.3CH.sub.2CH.sub.2PO.sub.3H.sub.2 Ethylphosphonic acid
CH.sub.3CH.sub.2PO.sub.3H.sub.2 Methylphosphonic acid
CH.sub.3PO.sub.3H.sub.2 tert-Butylphosphonic acid
(CH.sub.3).sub.3CPO.sub.3H.sub.2 Phenylphosphonic acid ##STR145##
3-Aminopropylphosphonic acid
NH.sub.2CH.sub.2CH.sub.2CH.sub.2PO.sub.3H.sub.2
(1-Aminopropyl)phosphonic acid ##STR146## Diethyl phosphoramidate
##STR147## 3-Aminopropyl(methyl)phosphinic acid, hydrochloride
##STR148## 4-Amino-1-butylphosphonic acid, disodium salt ##STR149##
1-(3-Phosphonopropyl)-benzimidazole, disodium salt ##STR150##
3-Dimethylamino-1-propylphosphonic acid, disodium salt ##STR151##
Diphenylamine-4-phosphonic acid, disodium salt ##STR152##
3-Amino-butylphosphonic acid, disodium salt ##STR153##
3-Amino-pentylphosphonic acid, disodium salt ##STR154##
3-Amino-hexylphosphonic acid, disodium salt ##STR155##
3-Amino-heptylphosphonic acid, disodium salt ##STR156##
3-Amino-octylphophonic acid, disodium salt ##STR157##
3-Amino-4-methyl-pentylphosphonic acid, disodium salt ##STR158##
3-Amino-3-methyl-butylphosphonic acid, disodium salt ##STR159##
3-Amino-3-phenyl-propylphosphonic acid, disodium salt ##STR160##
3-Amino-4-phenyl-butylphosphonic acid, disodium salt ##STR161##
3-Amino-4-phenyl-pentylphosphonic acid, disodium salt ##STR162##
3-Amino-3-phenyl-butylphosphonic acid, disodium salt ##STR163##
2-Amino-2-(2-phosphonoethyl)-1,3,4-tri- hydronaphthalene, disodium
salt ##STR164## 1-Amino-1-(2-phosphonoethyl)-cyclo- hexane,
disodium salt ##STR165## 2-(2-Amino-4-phos-
phonobutoxy)tetrahydropyran ##STR166##
3-Amino-4-hydroxy-butylphosphonic acid, disodium salt ##STR167##
3-Phosphonopropanesulfonic acid, trisodium salt ##STR168## Diethyl
2-pyrrolidinylphosphonate ##STR169## 2-Pyrrolidinylphosphonic acid,
disodium salt ##STR170## 1,1-Dioxo-2-(3-phosphonopropyl)-iso-
thiazoline, disodium salt ##STR171##
2-Deoxy-2-phosphonoacetylamino-D-glucose ##STR172##
3-Hydroxy-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt
##STR173## 3-Hydroxy-3-(3-pyridyl)propenyl-2-phos- phonic acid,
disodium salt ##STR174## 3-Hydroxy-3-(4-pyridyl)propenyl-2-phos-
phonic acid, disodium salt ##STR175##
3-Amino-3-(2-pyridyl)propenyl-2-phos- phonic acid, disodium salt
##STR176## 3-Amino-3-(3-pyridyl)propenyl-2-phos- phonic acid,
disodium salt ##STR177## 3-Amino-3-(4-pyridyl)propenyl-2-phos-
phonic acid, disodium salt ##STR178##
1,4-Diamino-1-(3-pyridyl)butyl-2-phos- phonic acid, disodium salt
##STR179## 1,4-Diamino-4-methyl-1-(3-py- ridyl)pentyl-2-phosphonic
acid, disodium salt ##STR180## 1,4-Diamino-4-methyl-1-(2-py-
ridyl)pentyl-2-phosphonic acid, disodium salt ##STR181##
1,4-Diamino-4-methyl-1-(4-py- ridyl)pentyl-2-phosphonic acid,
disodium salt ##STR182## 3-(2-Amino-4,5,7,8-tetrahydro-6H-thia-
zolo[4,5-d]azepin-6-yl)propyl- phosphonic acid, disodium salt
##STR183## 3-[6-Methoxy-2-(1,2,3,4-tetrahydro-
isoquinolinyl)]propylphosphonic acid, disodium salt ##STR184##
3-[8-Methoxy-2-(1,2,3,4-tetrahydro- isoquinolinyl)]propylphosphonic
acid, disodium salt ##STR185## 2-(3-Phosphonopropyl)-1,2,3,4-tetra-
hydro-9H-pyrido[3,4-b]indole, disodium salt ##STR186##
[0142] TABLE-US-00006 Phosphono Carbohydrates Name Structure
2-Deoxy-2-phosphonoacetyl- amino-D-glucose ##STR187##
2-Deoxy-2-thiophosphono- acetylamino-D-glucose ##STR188##
.beta.-D-Glucopyranosylmethyl- phosphonic acid, disodium salt
##STR189## .alpha.-D-Glucopyranosylmethyl- phosphonic acid,
disodium salt ##STR190## 6-Deoxy-6-C-phosphonomethyl-
D-glucono-.delta.-lactone, disodium salt ##STR191##
6-Deoxy-6-C-phosphonomethyl- D-glucose, disodium salt ##STR192##
4-Deoxy-4-C-phosphonomethyl- D-glucose, disodium salt ##STR193##
3-Deoxy-3-C-phosphonomethyl- D-glucose, disodium salt ##STR194##
1-Deoxy-N-phosphonoacetyl- nojirimycin, disodium salt ##STR195##
(1,5-Dideoxy-1,5-imino- .alpha.-D-glucopyranosyl)methyl phosphonic
acid, disodium salt ##STR196## 1,6-Dideoxy-6-C-phosphono-
methyl-nojirimycin, disodium salt ##STR197##
[0143] TABLE-US-00007 Thiophosphonate Derivatives Name Structure
Thiophosphonoformic acid, trisodium salt ##STR198##
Thiophosphonoacetic acid ##STR199## Thiophosphonoacetic acid,
trisodium salt ##STR200## Thiophosphonoacetic acid, triethyl ester
##STR201## Chloro(thiophosphono)acetic acid, trisodium salt
##STR202## Dichloro(thiophosphono)acetic acid, trisodium salt
##STR203## Thiophosphonomethylthiophosphonic acid, tetrasodium salt
##STR204## Phenylthiophosphinomethylthiophosphonic acid, trisodium
salt ##STR205##
3-[2-(1,2,3,4-Tetrahydroisoquinolinyl)]-1-propanethiophosphonic
acid, disodium salt ##STR206## Propylthiophosphonic acid ##STR207##
Ethylthiophosphonic acid ##STR208## Methylthiophosphonic acid
##STR209## tert-Butylthiophosphonic acid ##STR210##
2-Carboxyethylthiophosphonic acid ##STR211## Phenylthiophosphonic
acid ##STR212## 3-Aminopropylthiophosphonic acid ##STR213##
(dl)-2-Amino-3-thiophosphonopropionic acid ##STR214##
(1-Aminopropyl)thiophosphonic acid ##STR215##
(dl)-2-Amino-5-thiophosphonopentanoic acid ##STR216##
(S)-2-Amino-2-methyl-4-thiophosphonobutanoic acid ##STR217##
D-2-Amino-4-thiophosphonobutanoic acid ##STR218##
L-2-Amino-4-thiophosphonobutanoic acid ##STR219##
D-2-Amino-7-thiophosphonoheptanoic acid ##STR220##
L-2-Amino-7-thiophosphonoheptanoic acid ##STR221##
D-2-Amino-6-thiophosphonohexanoic acid ##STR222##
L-2-Amino-6-thiophosphonohexanoic acid ##STR223##
D-2-Amino-4-thiophosphonopentanoic acid ##STR224##
L-2-Amino-4-thiophosphonopentanoic acid ##STR225##
D-2-Amino-3-thiophosphonopropionic acid ##STR226##
L-2-Amino-3-thiophosphonopropionic acid ##STR227##
3-Aminopropyl(methyl)thiophosphinic acid, hydrochloride ##STR228##
(R)-3-(2-Carboxypiperazin-4-yl)-propyl-1-thiophosphonic acid
##STR229## L-4-[Difluoro(thiophosphono)methyl)]-phenylalanine
##STR230##
(R,E)-4-(3-Thiophosphonoprop-2-enyl)piperazine-2-carboxylic acid
##STR231## trans-L-4-Thiophosphonomethylproline, trisodium salt
##STR232## cis-L-4-Thiophosphonomethylproline, trisodium salt
##STR233## 4-Amino-1-butylthiophosphonic acid, disodium salt
##STR234## 1-(3-Thiophosphonopropyl)-benzimidazole, disodium salt
##STR235## 3-Dimethylamino-1-propylthiophosphonic acid, disodium
salt ##STR236## N,N-Diethylthiophosphonoacetamide, disodium salt
##STR237## Diphenylamine-4-thiophosphonic acid, disodium salt
##STR238## Selenophosphonoformic acid, trisodium salt ##STR239##
Selenophosphonoacetic acid, trisodium salt ##STR240##
D-2-Amino-3-selenophosphonopropanoic acid ##STR241##
L-2-Amino-3-selenophosphonopropanoic acid ##STR242##
D-2-Amino-4-selenophosphonobutanoic acid ##STR243##
L-2-Amino-4-selenophosphonobutanoic acid ##STR244##
N-Cyclohexylthiophosphonoacetamide, disodium salt ##STR245##
N-Cyclohexylselenophosphonoacetamide, disodium salt ##STR246##
N-Hydroxythiophosphonoacetamide, disodium salt ##STR247##
Thiophosphonoacetic hydrazide, disodium salt ##STR248##
N-Thiophosphonoacetyl-L-alanine, trisodium salt ##STR249##
N-Thiophosphonoacetyl-L-glycine, trisodium salt ##STR250##
N-(Thiophosphonoactyl)-L-asparagine-L-glycine, tetrasodium salt
##STR251## (s)-2-Pyrrolidinemethylthiophosphonic acid, disodium
salt ##STR252##
1,1-Dioxo-2-(3-thiophosphonopropyl)-isothiazolidine, disodium salt
##STR253## 2-Deoxy-2-thiophosphonoacetylamino-D-glucose ##STR254##
3-(2-Amino-4,5,7,8-tetrahydro-6H-thiazolo[4,5-d]azepin-6-yl)propyl-
thiophosphonic acid, disodium salt ##STR255##
1,1-Dioxo-2-(3-thiophosphonopropyl)-isothiazolidine, disodium salt
##STR256## 2-Deoxy-2-thiophosphonoacetylamino-D-glucose ##STR257##
3-(2-Amino-4,5,7,8-tetrahydro-6H-thiazolo[4,5-d]azepin-6-yl)propyl-
thiophosphonic acid, disodium salt ##STR258##
* * * * *