Lipophilic diesters of chelating agent for inhibition of enzyme activity
United States Patent: 7,799,831
Issued: September 21, 2010
Inventors: Striem; Sarina (Rehovot,
IL), Friedman; Jonathan Eduard (Rehovot, IL), Reznitsky-Cohen; Dalia (Nes-Ziona,
IL), Kozak; Alexander (Rehovot, IL)
Assignee: D-Pharm Ltd. (Kiryat
Weizmann Science Park Rehovot, IL)
Appl. No.: 10/529,028
Filed: March 16, 2003
PCT Filed: March 16, 2003
PCT No.: PCT/IL03/00225
371(c)(1),(2),(4) Date: March
PCT Pub. No.: WO2004/028443
PCT Pub. Date:
April 08, 2004
Executive MBA in Pharmaceutical Management, U. Colorado
The present invention relates to the use
of lipophilic diesters of the chelating agent 1,2-bis(2
aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) for inhibition of
proteolytic activities of certain metalloproteinases and of calpain. The
invention further relates to methods for preventing, treating or managing
MMP-related and calpain-related diseases or disorders in mammals
comprising administering to a mammal in need thereof, a pharmaceutical
composition comprising a therapeutically effective amount of said
lipophilic diesters of the chelating agent BAPTA.
Description of the
FIELD OF THE INVENTION
The present invention relates to the use of lipophilic diesters of the
chelating agent 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA)
for inhibition of proteolytic activities of certain metalloproteinases and
BACKGROUND OF THE INVENTION
Matrix metalloproteinases (MMPs) are extracellular zinc- and
calcium-dependent proteases, which are produced in a latent form and
require activation for catalytic activity. Activation occurs at the cell
surface and enables MMPs to degrade components of the extracellular matrix
(ECM) at specific sites in the membrane surroundings. The most studied
MMPs are the gelatinases, which include MMP-2 and MMP-9 that use gelatin,
Type IV collagen and fibronectin as preferred substrates. While
transcription of MMP-9 genes is transactivated by cytokines and growth
factors, MMP-2 is constitutively expressed and unresponsive to phorbol
ester and most cytokines. MMP-2 activation is regulated by MT1-MMP, which
is a membrane-anchored MMP. MMP-9 activation is regulated by a protease
cascade involving plasmin and stromelysin-1 (MMP-3).
Matrix metalloproteinases are responsible for much of the turnover of
matrix components and as such are involved in normal as well as in
pathological processes. MMPs have an important role in maintenance and
remodeling of membranes and ECM, for example, in breaking down the
extracellular matrix to allow cell growth and tissue remodeling during
development and recovery from injury. They also play a role in processes
such as ovulation, modulation of capillary permeability and in enabling
cell migration to a site of inflammation.
MMPs are involved in many pathological conditions. For example, they are
associated with pathogenic mechanisms in cancer such as invasion,
metastasis and angiogenesis [Reviewed by Foda and Zucker (2001) Drug
Discovery Today 6: 478-482]. MMPs play a part in progression of
inflammatory conditions and diseases involving degradation of
extracellular matrix, such as in stroke, hemorrhage, rheumatic diseases
(e.g. arthritis), Crohn's disease, asthma, and in cerebrovascular and
cardiovascular disorders [Mun-Bryce and Rosenberg (1998) J. Cerebral Blood
Flow Metabolism 18:1163-72; Yong et al. (1998) TINS 21:75-80; Lukes et al.
(1999) Mol. Neurobiol. 19:267-284]. Members of the MMP family have also
been implicated in neurological diseases and conditions as being involved
in demyelination and neuro-inflammatory processes. For example, MMPs have
been associated with brain damage and ischemia, Guillain-Barre, multiple
sclerosis, amyotrophic lateral sclerosis and Alzheimer's disease. The
current notion is that inflammation leads to the production of cytokines,
chemokines, growth factors and hormones that modulate MMPs production.
Activation of MMPs and plasminogen activators (PAs) is an important
regulatory step in the inflammatory response.
Members of another family of metalloproteinases, identified as "A
Disintegrin And Metalloproteases" (ADAM), have, like members of the MMP
family, multiple domains including a zinc-dependent catalytic domain and a
N-terminus pro-domain that is responsible for maintaining the enzyme in an
inactive state [Moss et al. (2001) Drug Discovery Today 6: 417-426]. It
was shown that members of the ADAM family are involved in several
different processing events including cleavage of substrates off the cell
membrane surface (a phenomenon termed "shedding"). One member of the ADAM
family is the TNF.alpha.-Converting Enzyme (TACE). TACE is found on the
cell surface where it processes the membrane-bound TNF.alpha., a
pro-inflammatory cytokine, to its mature soluble form. Soluble TNF.alpha.,
which is released in inflammatory conditions, can induce apoptosis. For
example, TNF induces secretion and activation of MMP-9 in macrophages and
glial cells and causes neuronal cell death in neuroinflammatory diseases
and following brain injuries. TNF has also been shown to play a role in
pathological conditions such as rheumatoid arthritis.
As potentially highly toxic proteolytic enzymes, the matrix
metalloproteinases are tightly regulated at multiple stages, as follows:
i) Gene transcription--most MMPs are not constitutively expressed, but
their transcription is controlled by various cytokines (e.g. IL-1, TNF)
and growth factors (e.g. TGF-.beta., retinoic acid, FGF).
ii) Pro-enzyme activation--MMPs are normally produced in a latent form
(pro-MMP) including a propeptide segment that generally must be removed to
activate the enzyme.
iii) Inhibition of enzyme activity--There are at least four endogenous MMP-inhibitors
known as tissue inhibitors of metalloproteinases (TIMPs), which bind to
the enzyme and block its activity. Another known natural inhibitor of MMPs
is the serum proteinase inhibitor .alpha.-macroglobulin.
Several synthetic inhibitors of MMPs have been described in various
publications in the scientific and patent literature. Currently known
inhibitors mainly include synthetic peptides and chelating agents
[reviewed by Woessner J F Jr. in Ann N.Y. Acad. Sci. (1999) 30:388-403].
Some synthetic inhibitors of the MMP active site are peptidomimetics based
on the sequence of peptides cleaved in collagen [Masui et al. (1977)
Biochem Med. 17:215-21). Peptidic agents based on conserved peptide
sequence derived from the pro-segment of human collagenase IV are
disclosed by Stelter-Stevenson et al. [Am J Med Sci. (1991) 302:163-70]
and in U.S. Pat. No. 5,270,447 to Liotta et al. Synthetic peptides
isolated from phage display peptide libraries and cyclic peptides with MMP
inhibitory activity are described by Koivunen et al. [Nat. Biothechnol
(1999) 17: 768-74].
N-hydroxyformamide peptidomimetics useful as TACE and MMPs inhibitors are
disclosed by Musso et al. [Bioorg Med Chem Lett (2001) 11: 2147-51].
Other polypeptides and peptoid compounds useful as metalloproteinase
inhibitors are disclosed in U.S. Pat. Nos. 4,263,293 and 4,297,275 to
Sundeen et al., in U.S. Pat. Nos. 4,371,465, 4,371,466 and 4,374,765 all
issued to McGregor, and in U.S. Patent Publication No. 2002/0090654 to
Langley et al.
Non-peptidic MMP-inhibitory compounds are disclosed in U.S. Pat. No.
4,950,755 to Witiak et al. and in U.S. Pat. No. 5,866,570 to Liang et al.
MMPs inhibitors comprising targeting moieties and chelators are disclosed
in International Patent Publication No. WO 01/60820 of Dupont
Pharmaceuticals Company, and in International Patent Publication No. WO
02/053173 to Kimberly-Clark Worldwide, Inc.
Matrix metalloproteinases as well as other members of the ADAM family are
inhibited by chelating agents. Most of these chelating agents are natural
and synthetic hydroxamate compounds and derivatives thereof such as
succinyl hydroxamate, sulfonamide hydroxamate etc. [reviewed by Woessner,
J. F. Jr. (1999) in Annals New York Academy of Sciences 30: 388-403]. For
example, the synthetic hydroxamates batimastat (BB-94; Invest New Drugs
(1996) 14:193-202) and its orally bioavailable analogue marimastat have
been shown to inhibit spread and growth of malignant tumors in animals.
These compounds are currently examined in advanced clinical trials.
Among the compounds that have been shown as MMPs inhibitors are also
antibiotics such as tetracyclines and their chemically modified analogs (Golub
et al. (1983) J Periodontal Res. 18:516-26; U.S. Pat. No. 4,704,383 to
McNamara et al.; U.S. Pat. No. 5,837,696 to Golub et al.].
Most of the above-mentioned agents are non-specific inhibitors of
metalloproteinases and other metal-ion dependent proteases.
Calpains are members of another family of proteases. These are cytosolic
enzymes which are calcium-dependent cysteine proteases. Calpains
predominantly exist within cells as inactive proenzymes and are converted
into their active forms in the presence of elevated intracellular calcium
levels. Upon binding of calcium, the precursor enzyme goes through a
self-digestion process that results in release of the activated calpain.
A wide range of proteins serves as substrates for calpain including
cytoskeletal, membrane and regulatory proteins. Calpain participates in a
number of normal cellular signal transduction systems as well as in
pathological conditions. For example, calpain activation has been
associated with ischemia and neuronal cell death such as those caused by
stroke and traumatic brain and spinal cord injuries [Bartus et al. (1995)
Neurol. Research 17:249-258]. Calpain proteolytic activity has also been
implicated in several neurodegeneration diseases and conditions, including
Alzheimer's Disease, Parkinson's disease, Huntington's disease and Pick's
Presently known natural and synthetic calpain inhibitors, including both
peptidic and non-peptide molecules, are reviewed by Wang and Yuen ["Calpain
inhibition: an overview of its therapeutic potential" in Trends Pharm.
Sci. (1994) 15, 412-419] and by Donkor ["A survey of calpain inhibitors"
in Curr. Med. Chem. (2000) 7:1171-88]. Known calpain inhibitors include
polypeptides which mimic peptide sequences of the natural inhibitors
calpastatin and kininogen [for review, see Wang and Yuen (1994) Trends
Pharm. Sci. (1994) 15, 412-419].
Compounds which are sulfonamide derivatives and ketone derivatives that
possess inhibitory activity against cysteine proteases are disclosed,
respectively, in U.S. Pat. Nos. 5,506,243 and 5,639,783 both to Ando et
al. Calpain inhibitors which are di-peptide alpha-keto esters, alpha-keto
amides and alpha-keto acids are described by Li et al. [J. Med Chem (1993)
36: 3472-80]. Several other classes of calpain inhibitors are disclosed by
Bartus et al. in International Patent Publication no. WO 92/11850.
Most commercially available calpain-inhibitors are compounds based on
peptide structures that interact with the substrate-binding site of the
enzyme. Many of these compounds are non-specific and inhibit a wide
variety of proteases in addition to calpain. Moreover, most of the known
inhibitors that were active in vitro, were found ineffective in inhibiting
calpain in-vivo, in particular in the CNS, as being poor membrane
permeants. Furthermore, almost all MMPs-inhibitors tested for treating
pathological inflammatory conditions or cancers failed in in-vivo clinical
Thus, there remains a long-felt need for effective, non-toxic agents which
are specific inhibitors of critical proteases such as the MMPs and calpain.
Stable lipophilic diesters of the divalent metal ion chelator 1,2-bis(2
aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) have been disclosed
in the International Patent Publication No. WO 99/16741 of the same
applicant. Also disclosed in this publication is the use of these
compounds in pharmaceutical compositions useful for treating diseases and
disorders related to excess of divalent metal ions. Among these diseases
and disorders are ischemia, stroke, epilepsy and neurodegenerative
diseases such as Alzheimer's disease and Parkinson's disease.
At that time, however, the mechanism by which these chelating agents exert
their neuroprotective effects has not been elucidated or disclosed. No
indication or suggestion for the cellular targets affected by these
chelators has been mentioned in the WO 99/16741 or any other publication.
SUMMARY OF THE INVENTION
It has now been found, in accordance with the present invention, that
certain diesters of the chelating agent 1,2-bis(2
aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (hereinafter denoted as
"DP-BAPTAs") are capable of inhibiting enzymatic activities of the
proteases matrix metalloproteinase (MMP), calpain and TNF.alpha.-Converting
Accordingly, the present invention provides, in one aspect, a method of
inhibiting protease activity, said protease being selected from
metalloproteinase and calpain, the method comprising exposing cells to
inhibiting amount of a compound of the general formula (I)
-- see Original Patent.
According to currently preferred
embodiments of the invention, the useful compounds for inhibiting the
MMP-9 activity are the following compounds and physiologically acceptable
1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-octoxyethyl acetate),
N,N'-diacetic acid (denoted herein DP-b99),
1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-octodecyloxyethyl acetate),
N,N'-diacetic acid (denoted herein DP-b109),
1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-benzyloxyethyl acetate), N,N'-acetic
acid (denoted herein DP-b440),
1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-dodecyloxyethyl acetate),
N,N'-diacetic acid (denoted herein DP-b460),
acetate], N,N'-diacetic acid (denoted herein DP-b458), and
Currently most preferred compounds for inhibiting calpain activity are
1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-octoxyethyl acetate),
N,N'-diacetic acid (DP-b99) and 1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-octodecyloxyethyl
acetate), N,N'-diacetic acid (DP-109) and physiologically acceptable salts
In another aspect of the invention, there are provided methods for
preventing, treating or managing diseases and pathological conditions
associated with damaging MMP and/or calpain activities. The methods
comprise administering to a mammal in need thereof, a pharmaceutical
composition containing as an active ingredient a therapeutically effective
amount of a compound of the above-mentioned general formula (I).
In yet another aspect of the invention, there is provided the use of a
compound of the general formula (I) for the preparation of a medicament
for inhibiting the activity of a protease selected from metalloproteinase,
calpain and TACE.
The present invention further provides methods and the use of the
compounds of the general formula (I) for the treatment of MMP- or calpain-related
diseases, disorders or conditions, which may be selected from the group
consisting of cancer (including metastasis cancer), angiogenesis-dependent
diseases (e.g. cancerous tumors, arthritis, psoriasis, macular
degeneration, chronic inflammation and diabetic retinopathy), ischemic or
hypoxic tissue damage, oxidative injury, stroke, trauma, inflammatory
conditions and diseases (e.g. arthritides, rheumatoid arthritis,
osteoarthritis, restenosis, asthma, psoriasis, systemic lupus
erythematosus, inflammatory bowel syndrome, Crohn's disease, gingivitis,
periodontitis, meningitis, tropical spastic paraparesis, sepsis, bullous
skin disorders, acne and inflammation due to infectious diseases),
atherosclerosis, thrombotic disorders, arthritis, osteoporosis, diabetes,
hemorrhage, autoimmune diseases, rheumatic diseases, ocular pathologies
and retinopathies (e.g. diabetic retinopathy, glaucoma, macular
degeneration, cataract, retinal detachment and retinal tears), burns,
chronic wounds (e.g. ulcers), neurological and neurodegenerative diseases
and disorders (e.g. multiple sclerosis (MS), Alzheimer's disease (AD),
motor neuron disease (MND), amyotrophic lateral sclerosis (ALS),
Guillain-Barre, Parkinson's disease, Huntington disease, Pick's disease,
dementia syndrome, vascular dementia, multiple infarct dementia,
HIV-induced neural disorders, brain ischemia (both global and focal
ischemia) and neuronal tissue trauma), migraine, cerebrovascular and
The methods of treatment in accordance with the invention may further
comprise treating the patient with additional therapeutic treatment which
may be carried out concurrently with, preceding or subsequent to the
administration of the pharmaceutical composition comprising a compound of
the general Formula (I).
It is important to note that not any chelator can inhibit the activity of
the tested proteases. In contrast to the effect of DP-BAPTA compounds, the
closely related known chelators, BAPTA and BAPTA-AM, at doses similar to
DP-BAPTA, did not inhibit MMP-9 activity. In fact, BAPTA-AM even slightly
enhanced the MMP-9 activity.
Another point to emphasize is that, under the experimental conditions
employed, the tested DP-BAPTAs inhibited calpain and MMP-9 proteolytic
activities, however no such effect could be demonstrated for the other
gelatinase tested, MMP-2.
DETAILED DESCRIPTION OF THE INVENTION
The synthesis and some utilization of stable lipophilic diesters of BAPTA
(DP-BAPTAs) have been disclosed in the International Patent Publication
No. WO 99/16741 of the same applicant, the teaching and disclosure of
which are expressly incorporated herein in their entirety by reference. In
the WO 99/16741 publication, the neuroprotective effects of DP-BAPTAs were
demonstrated in neuronal cell cultures in-vitro, and in ischemia model
systems in-vivo. However, the effect of the DP-BAPTA molecules on
activities of specific enzymes has not been taught or suggested in that or
any other publication. Accordingly, it was neither taught, recognized or
suspected that these compounds could be effectively use for the treatment
of MMP- and calpain-related diseases and disorders as disclosed in the
It is now disclosed, for the first time, that certain diesters of the
chelating agent BAPTA are capable of inhibiting the activity of calpain
and of certain proteases of the ADAM family, and in particular inhibiting
the activity of matrix metalloproteinase-9 (MMP-9).
The useful compounds in accordance with the invention are of the general
formula (I) as described above. It is to be understood that within the
scope of the invention are included also pharmaceutically acceptable salts
of the compounds of the general formula (I) including organic and
inorganic cations, as well as various solvates, including hydrates, and
other active forms of the compounds of the general formula (I).
Currently preferred compounds for inhibiting MMP or calpain activities are
diesters of BAPTA with alkyl chains comprising from around 8 to 20 carbon
atoms. The alkyl chains may be saturated or unsaturated alkyls including
one or more double bonds and/or a triple bond. According to preferred
embodiments of the invention, the alkyl chain is interrupted by from 1 to
3 oxygen atoms. According to most preferred embodiments, the R moiety of a
compound of the general formula (I) includes a monoalkyl ether of ethylene
glycols, preferably mono-, di- or tri-ethylene glycols.
The alkyl residue at position R of the compound of the general formula (I)
may consist of or include cyclic elements that may be aromatic or
non-aromatic ring structures. Preferably the cyclic elements have 5 or 6
Currently preferred cyclic R radicals comprise aromatic ring which is a
phenyl residue. Other currently preferred cyclic elements included at
position R are saturated or unsaturated cyclopentyl, cyclohexyl or
cycloheptyl. The cyclic elements may be directly linked to the carboxy
moiety of the compound of the general formula (I), or linked via a
saturated or unsaturated alkyl chain that may include one or more oxygen
and/or nitrogen atoms.
Preferably, the compound of the general formula (I) includes a monovalent
cation at position M. Suitable pharmaceutically acceptable cations may
include, but are not limited to, H.sup.+, Na.sup.+, Li.sup.+, K.sup.+,
NH.sub.4.sup.+ and mono-alkylammonium. Also divalent cations may be
included at position M. The choice of the preferred cation at position M
of the general formula (I) depends on the intended therapeutic use of the
compound, as well as on the specific formulation and route of
administration employed. A person skilled in the art will be able to
select the appropriate cation as required for the optimal pharmaceutical
compositions and way of administration chosen in each particular treated
One of the most preferred DP-BAPTA compounds is
1,2-bis(2-aminophenoxy)ethane, N,N'-di(2-benzyloxyethyl acetate), N,N'-acetic
acid (DP-b440), where the R moieties of the compound of the general
formula (I) include an alkylaryl moiety. This compound was disclosed
generally in the WO 99/16741 publication, but was not specifically claimed
and not individually tested.
It has now been shown by the inventors of the present invention that DP-BAPTAs
can attenuate or block both basal MMP-9 activity and TNF.alpha.- or PMA-induced
activation of MMP-9. DP-BAPTAs can also inhibit calpain activity. Hence,
DP-BAPTAs may be useful in reducing deleterious protease activities in
pathological conditions due, for example, to ischemia and inflammatory
responses. Accordingly, DP-BAPTA compounds may be useful in preventing,
treating or managing diseases and pathological conditions associated with
harmful activities of matrix metalloproteinases or calpains.
It is important to note that while MMP-9 activity was significantly
inhibited by the DP-BAPTA compounds, such inhibitory activity could not be
demonstrated with the related chelators tested, namely
1,2-bis(2-aminophenoxy)-ethane-N,N,N',N'-tetra-acetic acid (BAPTA) or
1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid acetoxymethyl
The DP-BAPTA compounds in accordance with the invention may be useful in
the treatment of a whole range of indications which involve degradation
processes carried out or mediated by MMPs, TACE or calpain. These
indications include, but are not limited to, diseases and conditions due
to neuronal ischemia (e.g. global brain ischemia and focal brain
ischemia), cardiac ischemia, trauma, stroke and inflammatory conditions
including neuroinflammatory diseases and disorders, rheumatic and
autoimmune diseases, neurological, cerebrovascular and cardiovascular
diseases and disorders. The DP-BAPTA compounds may also be useful in
compositions and methods for enhancing wound healing such as, for example,
in burns and in chronic wounds (e.g. ulcers).
A large amount of data has been accumulated which show MMP- and/or calpain-involvement
in progression of diseases and conditions where inflammatory processes are
implicated. These pathological conditions include, but are not limited to,
rheumatoid diseases (e.g. rheumatoid arthritis and osteoarthritis),
asthma, psoriasis, systemic lupus erythematosus, inflammatory bowel
syndrome, Crohn's disease, gingivitis, periodontitis, meningitis, tropical
spastic paraparesis, sepsis, bullous skin disorders, acne and inflammation
due to infectious diseases.
The infectious diseases may include, but are not limited to, infectious
diseases caused by any type of microorganism such as bacteria, fungi (e.g.
candidiatis, aspergilosis) and viruses (e.g. herpes viruses-related
disorders, HIV-related diseases), by parasites (e.g. malaria, amebiasis)
or by prions (e.g. Creutzfeld-Jacob Disease).
Inhibitors of MMPs or calpain can reduce proteolytic damage to tissues
such as that caused during inflammatory processes. For example, may limit
brain-blood-barrier (BBB) breakdown, inhibit neuroinflammation (e.g. as in
meningitis), reduce damage associated with brain or cardiac ischemic
injuries, and may diminish proteolytic effects caused by insults such as
oxidative stress, burns, infections and central (CNS) and peripheral
nervous system (PNS) injuries due to physical causes (e.g. trauma).
Elevated MMP or calpain activity has been linked to several
neurodegenerative diseases and conditions including, but not limited to,
multiple sclerosis (MS), motoneuron disease (MND), amyotrophic lateral
sclerosis (ALS), Alzheimer's disease, Guillain-Barre syndrome, Parkinson's
disease, Huntington's disease, Pick's disease, dementia syndrome, vascular
dementia, multiple infarct dementia, HIV-induced neural disorders, brain
ischemia (both global and focal ischemia) and neuronal tissue trauma.
MMPs have also been associated with pathological conditions such as
ischemic or hypoxic tissue damage, oxidative damage, osteoporosis,
hemorrhage, arterial restenosis, cardiovascular disorders (e.g. ischemic
myocardiac infarction) as well as with various ocular pathologies and
retinopathies including diabetic retinopathy, glaucoma, macular
degeneration, cataract, retinal detachment and retinal tears.
Cancer is another major disease where it has been shown that proteolytic
activities of metalloproteinases contribute to the progression of the
disease. MMPs are involved in spread of cancer, and in particular
facilitating the metastasis state of the disease. MMP-2 and MMP-9 are
involved in the breakdown of Type IV collagen, which is a major component
of basement membrane, and as such may be key factors in processes
involving membrane degradation, for example, in angiogenesis and in tumor
invasion and metastasis. Indeed, positive correlation has been found
between tumor progression and expression of members of the MMP family. For
example, increased expression of MMP-2 and MMP-9 genes has been associated
with malignancies of gliomas.
A number of factors are important in the progression of malignancies. One
of the crucial factors is angiogenesis, which is believed to be
fundamental for primary tumor growth, tumor progression and metastasis.
The first step in the mechanism of angiogenesis involves degradation of
basement membrane so to facilitate the growth of a new capillary sprout.
Thus, degradation and remodeling of the ECM are essential processes for
the mechanism of angiogenesis, and methods of inhibiting these processes
may be beneficial in blocking angiogenesis and hence diminishing
Angiogenesis is also important in a number of other pathological
processes, including arthritis, psoriasis, diabetic retinopathy, chronic
inflammation, scleroderma, hemangioma, retrolental fibroplasia and
abnormal capillary proliferation in hemophiliac joints, prolonged bleeding
etc. MMP-inhibitors are expected to be useful for the treatment of these
The inability to control metastasis presents a major problem, as
metastases are the leading cause of death in patients with cancer. To
date, there is no satisfactory treatment for preventing or limiting
metastasis growth. Thus, the use of the DP-BAPTA compounds in accordance
with the present invention for inhibiting MMPs, and in particular for
inhibiting the MMP-9 protease activity, may be beneficial in this respect.
The cancer subjected to treatment with DP-BAPTAs may include any type of
tumors and neoplastic growths that may be benign or malignant growths
including primary tumors as well as secondary tumors. The terms "cancer"
and "neoplastic growth" are interchangeably used in the specification and
claims and mean to cover all kinds of pathological uncontrolled cell
growths including invasive and non-invasive neoplasms, solid and non-solid
tumors, and including remote metastases.
The term "treatment" as used herein means to include therapeutic
procedures aiming at preventing, ameliorating, palliating, inhibiting or
delaying the onset and/or development and/or progression of a pathological
condition or improving its manifestations.
It should be understood that the therapeutic activity of the compounds of
the general formula (I), as disclosed and claimed herein, is irrespective
of the exact mechanism of action of these compounds and is not meant to be
limited to any particular mode of action by which these molecules exert
their beneficial effect(s).
In accordance with the methods of the invention, a pharmaceutical
composition comprising a therapeutically effective amount of a compound of
the general Formula (I) is administered to a patient in need thereof.
The administered pharmaceutical composition may include a compound(s) of
the general Formula (I) as the sole active ingredient, or may include said
compound(s) in combination with one or more additional agents known to be
effective in the treatment of a particular disease or disorder. The
compound(s) of the general Formula (I) and the additional therapeutically
active agent(s) may be included in the same pharmaceutical composition or
may be administered in separate compositions. Furthermore, the use of DP-BAPTA
in combination with another therapeutically active agent (or another
therapeutic means) may be concurrently or not. The methods of the
invention include administration of the DP-BAPTA compound(s) either at the
same time, preceding or following exposure to the additional therapeutic
agent or procedure.
Additional agents that may be used in combination with the DP-BAPTA
compounds may be therapeutic and prophylactic drugs, hormones, immuno-modifying
agents etc. and may include other chelating agents, proteins, peptides,
carbohydrates, lipidic molecules, DNA and RNA sequences etc.
These agents may be selected from, but are not limited to, anti-neoplastic,
anti-proliferative, anti-inflammatory, antibiotic, anti-viral,
anti-microbial, anti-mycotic, anti-allergic, cardiovascular agents, anti-convulsant,
anti-depressant, anti-psychotic, analgesic, neurological agents,
neuroprotective agents and bioactive peptides and proteins such as
neurotransmitters, immuno-modulators, growth factors, hormones, antibodies
For example, in the case of treating cancer, the DP-BAPTA compound(s) may
be used alone or in combination, concurrently or not, with additional
anti-cancer treatment. The additional anti-cancer treatment may include,
but is not limited to, chemotherapy, irradiation therapy, immunotherapy,
genetic therapy, surgery or any other anti-cancer treatment as known in
the art. The additional treatment may be carried out concurrently with or
consecutively to the administration of the compounds of the general
formula (I), namely the additional treatment may be applied concurrently
or successively, either preceding or subsequent to the administration of
the compound of the general formula (I). The time interval between the two
treatments and the overall regimen will be determined by a person skilled
in the art taking into account the specific treated disease and the
particular condition and response of the treated individual to the
Any anti-cancer drug that is suitable for use in chemotherapy procedures
may be applied in combination with the compound of the general formula
(I). Suitable anti-cancer drugs may include, but are not limited to,
alkaloids (e.g. taxol, vinblastine, vindesine and vincristine), alkylating
agents such as alkyl sulfonates, aziridines, ethylenimines,
methylmelamines, nitrogen mustards (e.g. cyclophosphamide) and
nitrosoureas, antibiotics and analogs (e.g. aclacinomycin, actinomycin,
anthramycin, daunorubicin and doxorubicin), antimetabolites such as folic
acid analogs (e.g. Tomudex.RTM.), purine and pyrimidine analogs and
platinum complexes (e.g. carboplatin, cisplatin, miboplatin and
The combination treatment with additional therapeutic procedures may be
beneficial also in treatment of other diseases and disorders. For example,
in treatment of inflammatory, neuro- or cardiovascular conditions where
the administration of the DP-BAPTA compounds may be in combination with
(either concurrently, preceding or subsequent to) surgery and/or treatment
with another medicament or therapeutic agent (e.g. antibiotics, antibodies
etc.) to remove or kill infectious agents or other pathogenic elements.
It will be readily apparent to those of ordinary skill in the art that a
large number of other beneficial drugs, reagents, means or procedures may
be useful in the treatment of particular pathological conditions.
Pharmaceutical compositions including these therapeutically effective
agents and methods applying them or other medical procedures are also
included within the scope of the invention as compositions and methods
useful in combination with the compounds of the general formula (I). The
exact protocol and the additional medicament or therapeutic procedure
used, will be determined by a person skilled in the art taking into
consideration the particulars of the specific medical condition treated,
e.g. the stage of the disease or disorder, its severity and progression,
as well as the condition of the patient.
The pharmaceutical compositions comprising the compound of the general
formula (I) may be in a liquid, aerosol or solid dosage form, and may be
formulated into any suitable formulation including, but not limited to,
solutions, suspensions, micelles, emulsions, microemulsions, aerosols,
ointments, gels, suppositories, capsules, tablets, and the like, as will
be required for the appropriate route of administration.
Any suitable route of administration is encompassed by the invention
including, but not being limited to, oral, intravenous, intraperitoneal,
intramuscular, subcutaneous, inhalation, intranasal, topical, rectal or
other known routes. In preferred embodiments, the useful pharmaceutical
compositions are orally or intravenously administered. The dose ranges are
those large enough to produce the desired proteinase inhibitory effect.
The dosing range varies with the specific DP-BAPTA used, the treated
pathological condition and the route of administration and is dependent on
the additional treatment procedure, if such additional treatment is
The dosage administered will also be dependent upon the age, sex, health,
weight of the recipient, concurrent treatment, if any, frequency of
treatment and the nature of the effect desired. The specific dosage,
regimen and means of administration will be determined by the attending
physician or other person skilled in the art.
Claim 1 of 8 Claims
1. A method for treating or managing a
metalloproteinase (MMP) or calpain related disease or disorder in a
mammal, the disease or disorder being glioma comprising administering to a
mammal in need thereof, a pharmaceutical composition comrising a
therapeutically effective amount of a compound of the general formula (I)
-- see Original Patent.
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