U.S. patent application number 11/527725 was filed with the patent office on 2008-03-27 for inhibition of cathepsin k activity and the treatment and prevention of disease.
This patent application is currently assigned to Sylvan Pharmaceuticals Pty Ltd.. Invention is credited to David Cullis-Hill, Robert Logan Hannon, Christopher Bond Little, Margaret Mary Smith.
Application Number | 20080076723 11/527725 |
Document ID | / |
Family ID | 39225780 |
Filed Date | 2008-03-27 |
United States Patent
Application |
20080076723 |
Kind Code |
A1 |
Cullis-Hill; David ; et
al. |
March 27, 2008 |
Inhibition of cathepsin K activity and the treatment and prevention
of disease
Abstract
The present invention provides a candidate compound that is
suitable for use in methods of treating animals (preferably
mammals) and in the preparation of a medicament, wherein the
candidate compound down regulates Cathepsin K activity.
Inventors: |
Cullis-Hill; David; (New
South Wales, AU) ; Hannon; Robert Logan; (New South
Wales, AU) ; Little; Christopher Bond; (New South
Wales, AU) ; Smith; Margaret Mary; (New South Wales,
AU) |
Correspondence
Address: |
BROWDY AND NEIMARK, P.L.L.C.;624 NINTH STREET, NW
SUITE 300
WASHINGTON
DC
20001-5303
US
|
Assignee: |
Sylvan Pharmaceuticals Pty
Ltd.
Bondi Junction
AU
|
Family ID: |
39225780 |
Appl. No.: |
11/527725 |
Filed: |
September 27, 2006 |
Current U.S.
Class: |
514/24 ;
514/54 |
Current CPC
Class: |
A61K 31/7024 20130101;
A61P 3/04 20180101; A61K 31/737 20130101 |
Class at
Publication: |
514/24 ;
514/54 |
International
Class: |
A61K 31/737 20060101
A61K031/737; A61K 31/7024 20060101 A61K031/7024 |
Claims
1. A method for treating or preventing bone resorption in a mammal
in need of such treatment, comprising the step of administering an
effective amount of an oversulfated polysaccharide.
2. The method according to claim 1 wherein bone resorption is
associated with bone metastases or multiple myeloma.
3. The method according to claim 1 wherein bone resorption is
associated with tooth root resorption.
4. The method according to claim 1 wherein bone resorption is
associated with osteolysis.
5. The method according to claim 4, wherein the osteolysis causes
loosening of a surgical prosthesis.
6. A method for treating or ameliorating a bone defect in a mammal
in need of such treatment, comprising the step of administering an
effective amount of an oversulfated polysaccharide.
7. The method according to claim 6 wherein the bone defect is a
post-traumatic injury.
8. The method according to claim 6 wherein the bone defect is a
delay or non-union of bones.
9. A method for treating or preventing tissue destruction
associated with inflammation of an airway or lung in a mammal in
need of such treatment, comprising the step of administering an
effective amount of an oversulfated polysaccharide.
10. The method according to claim 9, wherein the inflammation is
associated with chronic obstructive pulmonary disease, emphysema,
asthma, or lung fibrosis.
11. A method for treating obesity in a mammal in need of such
treatment comprising the step of administering an effective amount
of an oversulfated polysaccharide.
12. The method according to claim 1 wherein the oversulfated
polysaccharide is pentosan polysulfate.
13. The method according to claim 1 wherein the oversulfated
polysaccharide is a xylopyranose polysulfate.
14. The method according to claim 1 wherein the oversulfated
polysaccharide is a divalent salt.
15. The method according to claim 1 wherein the oversulfated
polysaccharide is calcium xylopyranose polysulfate.
Description
FIELD OF INVENTION
[0001] The invention relates to regulating the expression or
activity of cathepsin K in an animal in need thereof. More
particularly the invention relates to the treatment and prevention
of diseases in which the activity or expression of cathepsin K is
up regulated.
BACKGROUND OF THE INVENTION
[0002] Cathepsin K is a lysosomal enzyme with a typical acid pH
(5.5) optimum. The importance of this enzyme in intracellular
(lysosomal) collagenolytic activity has been demonstrated by the
accumulation of endocytosed collagen fibrils in fibroblasts treated
with a cathepsin K inhibitor in vitro. However, cathepsin K also
plays a significant role in extracellular type I collagenolysis in
the phagolysosome-like resorption compartment of active osteoclasts
in bone. Inhibition of cathepsin K in vitro and in vivo
significantly decreases osteoclastic bone resorption.
Pycnodysostosis (PD), a genetic disorder in humans characterised by
osteosclerosis and short stature is due to a deficiency in
cathepsin K activity. Individuals affected with PD have
significantly decreased bone collagen turnover, suggesting that
cathepsin K is the principle enzyme responsible for physiological
bone matrix turnover.
[0003] In pathological conditions such as in post-menopausal
osteoporosis (and ovariectomy models in rats) and other metabolic
bone disorders like Paget's disease, the excessive bone resorption
is also associated with collagen turnover by cathepsin K.
Furthermore, increased bone turnover in rheumatoid and
osteoarthritis is associated with elevated collagen cleavage by
cathepsin K.
[0004] In contrast, bone lysis and collagen release in cancer
metastasis to bone is thought to be predominantly due to enhanced
metallo matrix protein (MMP) activity. Similarly, while increased
cathepsin K expression in lysosomes of synovial fibroblasts and
chrondrocytes has been observed in arthritis, a direct role in
extracellular cartilage matrix degeneration has not been
demonstrated. In contrast, MMP-driven articular cartilage collagen
breakdown in vitro and in arthritic joints has been well
documented. Taken together, these data indicate that cathepsin K
plays a central role in type I collagenolysis and bone remodelling
in physiological and certain pathological conditions. Therefore,
mechanisms to inhibit cathepsin K activity represent excellent
targets for the treatment of excessive bone turnover.
[0005] Although once thought to reside exclusively in osteoclasts,
Cathepsin K expression has been discovered in a significant
fraction of human breast cancers, prostate cancer and associated
metastases.
[0006] Furthermore, activation of human pulmonary fibroblasts in
primary cell cultures has been found to lead to an increased
activity of Cathepsin K and to increase intracellular
collagenolytic activity suggesting that Cathepsin K plays a pivotal
role in lung matrix homeostasis under physiological and
pathological conditions.
SUMMARY OF THE INVENTION
[0007] In work leading to the present invention, the inventors
sought to provide an inhibitor of Cathepsin K activity that is
suitable for administration to animals and in particular mammals.
To that end, the inventors have surprisingly identified that an
oversulfated polysaccharide such as, for example pentosan
polysulfate (PPS) and more particularly xylopyranose polysulfate
(XPS) can significantly down regulate Cathepsin K activity or
expression, as well as inhibit osteoclast formation and osteoclast
function.
[0008] The present invention therefore provides a candidate
compound that is suitable for use in methods of treating animals
(preferably mammals) and in the preparation of a medicament,
wherein the candidate compound down regulates Cathepsin K
activity.
[0009] In one aspect the present invention provides methods for
treating or preventing bone resorption in an animal in need
thereof, the method comprising the step of administering an
effective amount of an oversulfated polysaccharide.
[0010] In a preferred embodiment the oversulfated polysaccharide is
pentosan polysulfate, more preferably xylopyranose polysulfate.
Preferably the pentosan polysulfate is calcium pentosan
polysulfate, more preferably calcium xylopyranose polysulfate.
[0011] As used herein, the term "bone resorption" refers to a loss
of bone through increased breakdown via osteoclasts or other
mechanism causing a reduction in bone mass. Throughout this
specification the word "comprise", or variations such as
"comprises" or "comprising", will be understood to imply the
inclusion of a stated element, integer or step, or group of
elements, integers or steps, but not the exclusion of any other
element, integer or step, or group of elements, integers or
steps.
[0012] The Cathepsin K inhibitors identified by the present
inventors are particularly useful in pathological processes
involving excess osteoclast activation and bone resorption, such as
bone metastases or multiple myeloma.
[0013] Accordingly, the present invention provides a method of
treating or preventing bone resorption associated with bone
metastases or multiple myeloma in a mammal in need of such
treatment, comprising the step of administering an effective amount
of an oversulfated polysaccharide.
[0014] The Cathepsin K inhibitors identified by the present
inventors are also useful in other processes involving bone matrix
resorption. For example, the Cathepsin K inhibitors are useful in
the treatment and prevention of tooth root resorption, and
osteolysis associated with surgical implants and joint
replacement.
[0015] Accordingly the present invention provides a method for
treating or preventing bone resorption associated with tooth root
resorption in a mammal in need of such treatment, comprising the
step of administering an effective amount of an oversulfated
polysaccharide.
[0016] Resorption of a part of the root of a tooth may be either
internal (pulpal) or external.
[0017] The present invention further provides a method for treating
or preventing excess bone resorption associated with osteolysis in
a mammal in need of such treatment, comprising the step of
administering an effective amount of an oversulfated
polysaccharide.
[0018] Osteolysis refers to an active resorption or dissolution of
bone tissue as part of an ongoing disease process. While bone
resorption is commonly associated with many diseases, the term
osteolysis generally refers to a problem common to artificial joint
replacements such as total hip replacements, total knee
replacements and total shoulder replacements. In a total hip
replacement, for example, the particles worn off the gliding
surface of the ball and socket joint often cause osteolysis. As the
body attempts to clean up these loose particles of plastic or
metal, the bone grows away from the implant, causing it to
loosen.
[0019] The present invention further provides a candidate compound
suitable for use in the treatment of or amelioration of bone
defects. Defects include, for example, post traumatic injuries such
as fractures or defects of the bone after a bone resection or
removal of a growth (such as a cyst) in the bone. In one embodiment
the candidate compound is useful in the treatment of delayed
healing of bone fractures and infection associated delayed healing
or non-unions of fractures.
[0020] In a preferred embodiment the oversulfated polysaccharide is
pentosan polysulfate, more preferably calcium pentosan polysulfate.
In a more preferred embodiment the oversulfated polysaccharide is
xylopyranose polysulfate, more preferably calcium xylopyranose
polysulfate.
[0021] In another aspect the present invention provides methods for
treating or preventing tissue destruction associated with
inflammation of an airway or lung in an animal in need thereof, the
method comprising the step of administering an effective amount of
an oversulfated polysaccharide.
[0022] In a preferred embodiment the oversulfated polysaccharide is
pentosan polysulfate, more preferably calcium pentosan polysulfate.
In a more preferred embodiment the oversulfated polysaccharide is
xylopyranose polysulfate, more preferably calcium xylopyranose
polysulfate.
[0023] Inflammation of a lung or airway usually involves
infiltration of one or more leucocytes including neutrophils,
eosinophils, lymphocytes and monocytes but can also involve
macrophages, mast cells or basophils. In inflammation, there is an
expression of a wide array of catabolic enzymes including
proteases. Those lesions expressing the protease Cathepsin K have a
uniquely enhanced collagen cleaving activity which opens up further
targets in tissues for other enzymes and causes tissue destruction.
Conditions which involve inflammation include, for example, chronic
obstructive pulmonary diseases with or without acute infections or
worsening of inflammation, lung fibrosis, allergic diseases and
asthma. This also includes inflammation in the airways including
upper airways caused by allergy or infection or other diseases.
Accordingly the present invention provides a method for treating or
preventing tissue destruction associated with chronic obstructive
pulmonary disease (COPD) in a mammal in need of such treatment,
comprising the step of administering an effective amount of an
oversulfated polysaccharide. In one embodiment the COPD is related
to emphysema.
[0024] In another embodiment the present invention provides a
method for treating or preventing tissue destruction associated
with lung fibrosis in a mammal in need of such treatment,
comprising the step of administering an effective amount of an
oversulfated polysaccharide.
[0025] In yet another embodiment the present invention provides a
method for treating or preventing obesity in a mammal in need of
such treatment, comprising the step of administering an effective
amount of an oversulfated polysaccharide.
[0026] In a preferred embodiment the oversulfated polysaccharide is
pentosan polysulfate, more preferably calcium pentosan polysulfate.
In a more preferred embodiment the oversulfated polysaccharide is
xylopyranose polysulfate, more preferably calcium xylopyranose
polysulfate.
[0027] The methods of the present invention comprise administration
of an effective amount of the candidate compound to the individual
in need thereof. The present invention further provides for the use
of the candidate compound alone or together with a suitable
excipient in the preparation of a medicament.
[0028] The term "effective amount" refers to a therapeutically
effective amount and refers to an amount of the compound to treat,
ameliorate, or prevent a desired disease or condition, or to
exhibit a detectable therapeutic or preventative effect. The effect
can be detected by, for example, chemical markers. Therapeutic
effects also include reduction in physical symptoms, such as
decreased bone resorption decreased tissue destruction and even
improved condition of the bone or tissue. The precise effective
amount for a situation is determined by routine experimentation and
is within the judgement of the clinician.
[0029] The compound is administered preferably in an amount such as
to produce a concentration of the compound in the blood of 0.01 to
100 micrograms/ml plasma, for example 0.1 to 50 micrograms per ml
plasma. Typically, administration of about 0.1-30 mg/kg body weight
of the compound will produce a plasma concentration in the range of
0.1-100 micrograms/ml. The compound can be administered in a single
dose or in multi-dosage form, at one time or intermittently to
reduce any effects of toxicity.
[0030] Any administration known in the art may be used. In one
preferred embodiment the candidate compound is administered
systematically (by injection). One preferred systemic dose is about
1-5 mg/kg, more preferably 2 mg/kg body weight.
[0031] In another embodiment the candidate compound is administered
orally. A preferred oral dose is about 10-30 mg/kg, more preferably
about 20 mg/kg body weight.
[0032] It will be appreciated by persons skilled in the art that
numerous variations and/or modifications may be made to the
invention as shown in the specific embodiments without departing
from the spirit or scope of the invention as broadly described. The
present embodiments are, therefore, to be considered in all
respects as illustrative and not restrictive.
BRIEF DESCRIPTION OF THE FIGURES
[0033] FIG. 1 is a bar graph for the results of dose response of
CaXPS on Cathepsin K mediated release of bone collagen
(Mean.+-.SD). *=p<0.05 vs no CaXPS.
[0034] FIG. 2 is a bar graph for the results of multinucleated cell
count of RAW 264.7 cell cultures.+-.RANKL administration on day 0
(Mean.+-.SD of cells with .gtoreq.2 nuclei). *=p<0.05 vs Day 2.
***=p<0.0005 vs Day 2. #=p<0.05, -vs+RANKL.
[0035] FIGS. 3A(i), (ii) and B(i), (ii) are micrographs of H&E
Staining of RAW cells for counts (200.times. magnification). Arrows
indicate multinucleated OCL cells.
[0036] FIGS. 4 A and B are bar graphs for the results of the effect
of CaXPS on the percentage of cells with (A) 2-3 nuclei and (B)
>4 nuclei (OCL cells) at day 4 of RAW 264.7 cell
cultures.+-.RANKL (Mean.+-.SD). *=p<0.05, -vs+RANKL.
**=p<0.005, -vs+RANKL. #=p<0.05 vs no CaXPS.
[0037] FIGS. 5A and B are bar graphs for the results of dose
response of CaXPS on (A) Cathepsin K and (B) MMP9 gene expression
by RAW 264.7 cells.+-.RANKL at day 4 (Mean.+-.SD). #=p<0.05 vs
no CaXPS. *=p<0.05, -vs+RANKL. **=p<0.005, -vs+RANKL.
[0038] FIG. 6 is a .sup.13C-NMR spectrum of the .sup.13C-NMR data,
the position of the zinc [A] and calcium [B] atoms in the complexes
formed with Pentosan polysulfate are shown as black spheres. Note
that in the zinc complex [A] the metal resides in the cleft between
the pentosan rings, whereas in [B] the calcium occupies a position
across the C-3 and C-5 positions of the sugar ring and close to the
sulphate groups.
DETAILED DESCRIPTION
Polysulfated Polysaccharides
[0039] Xylopyranose polysulfate (XPS) which is a semi-synthetic
derivative of beechwood, is one example of an oversulfated
polysaccharide useful in the present invention.
[0040] Pentosan polysulfates (PPS) and, more particularly,
xylopyranose polysulfates are available as an alkali metal salt or
alkaline earth metal salt, for example, comprising calcium or
sodium salt, or transition metals such as copper and zinc and noble
metals such as platinum. Accordingly, the particular complexing
ions may be selected from the group consisting of the alkali
metals, e.g. Na+ and K+, alkaline earth metals, e.g. Ca2+, Zn2+,
Mg2+, Ba2+, as well as Ag+, Pb2+, Cu2+, Au2+, Pd2+, Pd4+, Pd4+,
Pd2+, trivalent metal ions, and quaternary ammonium compound
complexes. Examples of the latter compound are pyridinium chloride,
tetraalkyl ammonium chloride, chorine chloride, cetylpyridinium
chloride, N-cetyl-N,N,N-trialkylammonium chloride or their
derivatives. The most preferred of these are the divalent alkaline
earth metals, preferably calcium, and magnesium and most preferable
is the calcium complex. Preparation of the polysulfate
polysaccharide-metal complexes is described in detail in U.S. Pat.
No. 5,668,116, the entire disclosure of which is incorporated
herein by reference.
[0041] Other polysulfated polysaccharides included within the scope
of the invention are, for example, polysulfated dextran and
derivatives thereof, polysulfated cyclodextrin, sulfated heparin,
sulfated mannose and mannose derivatives, xylan, polysulfated
chondroitin, dermatan and hyaluronic acid. Further examples are
polysulfated polysaccharide derivatives of homopolysaccharides or
heteropolysaccharides which can be linear or branched. As described
above, complexes are also formed between these polysulfated
polysaccharides and multivalent metal ions, Ag+ and Au+, and
quaternary ammonium compound complexes.
[0042] The sugars may come from but are not limited to pentoses or
hexoses such as galactose, mannose, glucose, rhanose, fructose,
sorbose, xylose, D-arabinose, ribose, L-arabinose, glucuronic acid
and their derivatives.
[0043] The term oversulfated refers to the compound having a
sulfate group attached to all oxygen sites that are available for
sulfation. For example, PPS contains approximately two sulfate
groups per carbohydrate monomer. Due to uronic acid side groups on
PPS, the degree of sulfation on PPS is approximately 1.8.
[0044] Sulphation of polysaccharides is described in detail in U.S.
Pat. No. 5,668,116, the entire disclosure of which is incorporated
herein by reference.
[0045] The compound may be administered to the patient alone or in
combination with a pharmaceutically acceptable carrier.
Method of Administration
[0046] The administration according to the use and method of the
present invention may be any administration known in the art as may
easily be recognised by the skilled person according to the
individual situation. Accordingly, the administration may be
selected from systemic administration; injection into tissue or
into a body cavity including joints; implantation into tissue or
into a body cavity; topical application to the skin or to any
gastrointestinal surface or to a mucosal surface including the
lining of body cavities. The administration may be selected from
parenteral administration, including intraperitoneal
administration, intrathecal administration, systemic
administration, local administration, topical administration,
transmucosal administration, transdermal administration and oral
administration.
[0047] One preferred method of administration according to the
method of the invention is by the oral route. Another preferred
method of administration is by infiltration for treatments around a
tooth root, prosthesis or bone defects. Infiltration can include
either topical application into and around the lesion or
alternatively local injection into and around the lesion. In yet
another embodiment the preferred mode of administration is
systemic, i.e. by iv, im or subcutaneous injection. For injection,
the carrier would typically be for example normal sterile
physiological saline.
Pharmaceutical Formulations and Compositions
[0048] These are intended to include pharmaceutical compositions,
cosmetics and cosmeceuticals.
[0049] For the administration to an individual (an animal or a
human) the substances are preferably formulated into a
pharmaceutical composition containing the target compound and,
optionally, one or more pharmaceutically acceptable excipients.
[0050] The compositions may be in the form of a solid, semi-solid
or fluid composition such as, but not limited to, for example,
bioabsorbable patches, drenches, dressings, hydrogel dressings,
hydrocolloid dressings, films, foams, sheets, bandages, plasters,
delivery devices, implants, powders, granules, granulates,
capsules, agarose or chitosan beads, tablets, pills, pellets,
microcapsules, microspheres, nanoparticles, sprays, aerosols,
inhalation devices, gels, hydrogels, pastes, ointments, creams,
soaps, suppositories, vagitories, tooth paste, solutions,
dispersions, suspensions, emulsions, mixtures, lotions, mouthwash,
shampoos, enemas.
[0051] A pharmaceutical composition comprising an active substance
serves as a drug delivery system. In the present context the term
"drug delivery system" denotes a pharmaceutical composition (a
pharmaceutical formulation or a dosage form) which upon
administration presents the active substance to the body of a human
or an animal and if necessary facilitates the presentation of the
active substance to the appropriate site of treatment. Thus, the
term "drug delivery system" embraces plain pharmaceutical
compositions such as, e.g., creams, ointments, liquids, powders,
tablets, etc. as well as more sophisticated formulations such as
sprays, plasters, bandages, dressings, devices, etc.
[0052] The compositions may be formulated according to conventional
pharmaceutical practice, see, e.g., "Remington: The science and
practice of pharmacy" 20th ed. Mack Publishing, Easton Pa., 2000
ISBN 0-912734-04-3 and "Encyclopedia of Pharmaceutical Technology",
edited by Swarbrick, J. & J. C. Boylan, Marcel Dekker, Inc.,
New York, 1988 ISBN 0-8247-2800-9.
[0053] Apart from the candidate compound, a pharmaceutical
composition for use according to the invention may comprise
pharmaceutically or cosmetically acceptable excipients.
[0054] The choice of pharmaceutically acceptable excipients in a
composition for use according to the invention and the optimum
concentration thereof cannot generally be predicted and must be
determined on the basis of an experimental determination thereof.
Also whether a pharmaceutically acceptable excipient is suitable
for use in a pharmaceutical composition is generally dependent on
which kind of dosage form is chosen. However, a person skilled in
the art of pharmaceutical formulation can find guidance in e.g.,
"Remington: The science and practice of pharmacy" 20th ed. Mack
Publishing, Easton Pa., 2000 ISBN 0-912734-04-3.
[0055] A pharmaceutically acceptable excipient is a substance,
which is substantially harmless to the individual to which the
composition will be administered. Such an excipient normally
fulfils the requirements given by the national drug agencies.
Official pharmacopoeias such as the British Pharmacopeia, the
United States of America Pharmacopeia and the European Pharmacopeia
set standards for well-known pharmaceutically acceptable
excipients.
[0056] In the following is given a review on relevant
pharmaceutical compositions for use according to the invention. The
review is based on the particular route of administration. However,
it is appreciated that in those cases where a pharmaceutically
acceptable excipient may be employed in different dosage forms or
compositions, the application of a particular pharmaceutically
acceptable excipient is not limited to a particular dosage form or
of a particular function of the excipient.
Parenteral Compositions
[0057] For systemic application, the compositions according to the
invention may contain conventionally non-toxic pharmaceutically
acceptable carriers and excipients for example including
microspheres and liposomes.
[0058] The compositions for use according to the invention include
all kinds of solid, semisolid and fluid compositions. Compositions
of particular relevance are e.g. solutions, suspensions, emulsions,
gels, implantation tablets and implants.
[0059] The pharmaceutically acceptable excipients may include
solvents, buffering agents, preservatives, humectants, chelating
agents, antioxidants, stabilizers, emulsifying agents, suspending
agents, gel-forming agents, diluents, disintegrating agents,
binding agents, lubricants and wetting agents. For examples of the
different agents see below.
Topical, Transmucosal and Trans-Dermal Compositions:
[0060] For application to the mucosa or the skin, the compositions
for use according to the invention may contain conventionally
non-toxic pharmaceutically acceptable carriers and excipients
including microspheres and liposomes.
[0061] The compositions for use according to the invention include
all kinds of solid, semi-solid and fluid compositions. Compositions
of particular relevance are e.g. pastes, ointments, hydrophilic
ointments, creams, gels, hydrogels, solutions, emulsions,
suspensions, lotions, liniments, resoriblets, suppositories, enema,
pessaries, moulded pessaries, vaginal capsules, vaginal tablets,
shampoos, jellies, soaps, sticks, sprays, powders, films, foams,
pads, sponges (e.g. collagen sponges), pads, dressings (such as,
e.g., absorbent wound dressings), drenches, bandages, plasters and
transdermal delivery systems.
[0062] The pharmaceutically acceptable excipients may include
solvents, buffering agents, preservatives, humectants, chelating
agents, antioxidants, stabilizers, emulsifying agents, suspending
agents, gel-forming agents, ointment bases, suppository bases,
penetration enhancers, perfumes, skin protective agents, diluents,
disintegrating agents, binding agents, lubricants and wetting
agents. For examples of the different agents see below.
[0063] For application to the mucosa or the skin, the compositions
for use according to the invention may contain conventionally
non-toxic pharmaceutically acceptable carriers and excipients
including microspheres and liposomes.
Oral Compositions:
[0064] The composition for use according to the invention includes
all kinds of solid, semi-solid and fluid compositions. Compositions
of particular relevance are e.g. solutions, suspensions, emulsions,
uncoated tablets, modified-release tablets, gastro-resistant
tablets, orodispersible tablets, effervescent tablets, chewable
tablets, soft capsules, hard capsules, modified release capsules,
gastro-resistant capsules, uncoated granules, effervescent
granules, granules for the preparation of liquids for oral use,
coated granules, gastro-resistant granules, modified-release
granules, powders for oral administration and powders for the
preparation of liquids for oral use.
[0065] The pharmaceutically acceptable excipients may include
solvents, buffering agents, preservatives, humectants, chelating
agents, antioxidants, stabilizers, emulsifying agents, suspending
agents, gel-forming agents, diluents, disintegrating agents,
binding agents, lubricants, coating agents and wetting agents. For
examples of the different agents see below.
Examples of Various Agents:
[0066] Examples of solvents are but not limited to water, alcohols,
vegetable or marine oils (e.g. edible oils like almond oil, castor
oil, cacao butter, coconut oil, corn oil, cottonseed oil, linseed
oil, olive oil, palm oil, peanut oil, poppyseed oil, rapeseed oil,
sesame oil, soybean oil, sunflower oil, and teaseed oil), mineral
oils, fatty oils, liquid paraffin, polyethylene glycols, propylene
glycols, glycerol, liquid polyalkylsolixanes, and mixtures
thereof.
[0067] Examples of buffering agents are but not limited to citric
acid, acetic acid, tartaric acid, lactic acid, hydrogenphosphoric
acid, diethylamine etc.
[0068] Examples of preservatives for use in compositions are but
not limited to parabens, such as methyl, ethyl, propyl
p-hydroxybenzoate, butylparaben, isobutylparaben, isopropylparaben,
potassium sorbate, sorbic acid, benzoic acid, methyl benzoate,
phenoxyethanol, bronopol, bronidox, MDM hydantoin, iodopropynyl
butylcarbamate, EDTA, benzalconium chloride, and benzylalcohol, or
mixtures of preservatives.
[0069] Examples of humectants are but not limited to glycerine,
propylene glycol, sorbitol, lactic acid, urea, and mixtures
thereof.
[0070] Examples of chelating agents are but not limited to sodium
EDTA and citric acid.
[0071] Examples of antioxidants are but not limited to butylated
hydroxy anisole (BHA), ascorbic acid and derivatives thereof,
tocopherol and derivatives thereof, cysteine, and mixtures
thereof.
[0072] Examples of emulsifying agents are but not limited to
naturally occurring gums, e.g. gum acacia or gum tragacanth;
naturally occurring phosphatides, e.g. soybean lecithin; sorbitan
monooleate derivatives; wool fats; wool alcohols; sorbitan esters;
monoglycerides; fatty alcohols; fatty acid esters (e.g.
triglycerides of fatty acids); and mixtures thereof.
[0073] Examples of suspending agents are but not limited to
celluloses and cellulose derivatives such as, e.g.
carboxymethylcellulose, hydroxyethylcellulose,
hydroxypropylcellulose, hydroxypropylmethylcellulose, carraghenan,
acacia gum, arabic gum, and mixtures thereof.
[0074] Examples of gel bases and viscosity-increasing are but not
limited to liquid paraffin, polyethylene, fatty oils, colloidal
silica or aluminium, zinc soaps, glycerol, propylene glycol,
tragacanth, carboxyvinyl polymers, magnesium-aluminium silicates,
Carbopol.RTM., hydrophilic polymers such as, e.g. starch or
cellulose derivatives, water-swellable hydrocolloids, carragenans,
hyaluronates (e.g. hyaluronate gel optionally containing sodium
chloride), and alginates including propylene glycol alginate.
[0075] Examples of ointment bases are but not limited to beeswax,
paraffin, cetanol, cetyl palmitate, vegetable oils, sorbitan esters
of fatty acids (Span), polyethylene glycols, and condensation
products between sorbitan esters of fatty acids and ethylene oxide,
e.g. polyoxyethylene sorbitan monooleate (Tween).
[0076] Examples of hydrophobic ointment bases are but not limited
to paraffins, vegetable oils, animal fats, synthetic glycerides,
waxes, lanolin, and liquid polyalkylsiloxanes.
[0077] Examples of hydrophilic ointment bases are but not limited
to solid macrogols (polyethylene glycols).
[0078] Examples of powder components are but not limited to
alginate, collagen, lactose, powder which is able to form a gel
when applied to a wound (absorbs liquid/wound exudates).
[0079] Examples of diluents and disintegrating agents are but not
limited to lactose, saccharise, emdex, calcium phosphates, calcium
carbonate, calcium sulphate, mannitol, starches and
microcrystalline cellulose.
[0080] Examples of binding agents are but not limited to
saccharise, sorbitol, gum acacia, sodium alginate, gelatine,
starches, cellulose, sodium coboxymethylcellulose, methylcellulose,
hydroxypropylcellulose, polyvinylpyrrolidone and
polyetyleneglycol.
[0081] Examples of wetting agents are but not limited to sodium
laurylsulphate and polysorbate 80.
[0082] Examples of lubricants are but not limited to talcum,
magnesium stearate, calcium stearate, silicium oxide, precirol and
polyethyleneglycol.
[0083] Examples of coating agents are but not limited to
hydroxypropylcellulose, hydroxypropylmethylcellulose,
polyvinylpropylidone, ethylcellulose and polymethylacrylates.
[0084] Examples of suppository bases are but not limited to oleum
cacao, adeps solidus and polyethylenglycols.
[0085] The candidate compound is present in the medicament in an
amount of 0.001-99%, typically 0.01-75%, more preferably 0.1-20%,
and especially 1-10% by weight of the medicament.
The Role of Cathepsin K in Disease
Cathepsin K is Expressed in Human Breast Carcinoma
[0086] In patients with bone metastases, bone loss is the
consequence of a dissociated process combining excessive bone
resorption and inhibited bone formation. Invading cells in bone
metastases of breast cancer have been found to express Cathepsin K.
Immunolocalization of Cathepsin K in breast tumor bone metastasis
has revealed that the invading breast cancer cells expressed this
protease, albeit at a lower intensity than in osteoclasts.
Expression of Cathepsin K mRNA has been confirmed by reverse
transcription PCR and Southern analysis in a number of human breast
cancer cell lines and in primary human breast tumors and their
metastasis. Thus, the present invention provides a novel candidate
compound suitable for use in a medicament for preventing the
invasive potential of breast cancer cells, in particular those that
degrade bone matrix proteins, and treating or preventing bone
resorption associated with breast cancer.
Prostate Cancer
[0087] Prostate cancer is often also associated with bone
metastasis, which cause much of the morbidity associated with
prostate cancer. Prostate cancer bone metastasis exhibit increases
in both bone formation and resorption. It has been found that
Cathepsin K is expressed in bone metastasis at a significantly
higher rate that in primary prostate cancer. Lesions associated
with prostate cancer generally exhibit increased bone formation and
resorption. Increased bone resorption may release factors from the
extra cellular matrix that contribute to tumor growth. Prostate
cancer expressed Cathepsin K is postulated to contribute to the
invasive potential of prostate cancer, while increased expression
in bone metastasis is consistent with a role in matrix degradation.
Therefore, the present invention provides a novel candidate
compound suitable for use in a medicament for treating or
preventing the invasive potential of prostate cancer, and in
particular treating or preventing bone resorption associated with
prostate cancer.
Lung Cancer
[0088] In study a of osteoclastic functions in osteolytic
metastasis of lung cancers, bone resorping osteoclasts were
reactive for both cathepsin K and MMP-9, whereas the osteoclasts in
the stromal tissue of the tumour nests showed only MMP-9
immunoreactivity. Osteoclasts appeared to easily migrate in the
stromal tissue and to be involved in the alteration of
microenvironment of osteolytic metastasis by extracellular matrix
degradation, as well as bone resorption.
Giant Cell Tumor
[0089] Giant cell tumor (GCT) of bone is a neoplasm of bone
characterized by a localized osteolytic lesion. It has been found
that cathepsin K is the principal protease in giant cell tumor of
bone, and it is suggested that osteoclast-like giant cells are
responsible for osteolysis. Inhibition of cathepsin K or its
associated proton-pump would provide a new therapeutic opportunity
for GCT, and accordingly the present invention provides a novel
method for treating or preventing GST of bone.
Chordoma
[0090] Invasive growth of chordoma is accompanied by severe
destruction of adjacent bone tissue, a fact that requires high
proteolytic activity at the tumor invasion fronts. Enzyme
histochemistry indicates a strong cell-associated cathepsin K
activity in invasive tumor components. Studies have shown that the
significant expression and activity of cathepsin K in chordoma and
implicates an important role of the protease in the infiltrative
growth of the tumor.
Thyroid Carcinoma
[0091] Expression of Cathepsin K in multinucleated giant cells
(MGC) of the anaplastic carcinoma of the thyroid gland is
postulated to contribute to the invasive behaviour of this tumour
thus promoting metastatic ability and destruction of the
cartilaginous trachea. The present invention provides a novel
method for treating or preventing MGC of the carcinoma of the
thyroid gland.
Multiple Myeloma
[0092] Multiple myeloma is characterized by the accumulation of
clonal malignant plasma cells in the bone marrow, which stimulates
bone destruction by osteoclasts and reduces bone formation by
osteoblasts. A challenge for treating multiple myeloma is
discovering drugs targeting not only myeloma cells but also
osteoclasts and osteoblasts. In cultures of human primary
monocytes, it has been found that there is an up-regulation of
cathepsin K gene expression. Cathepsin K is considered to be a
target for treating multiple myeloma and its effects on bone.
Tooth Root Loosening
[0093] Investigations of the expression and localisation of
Cathepsin K in root-resorping tissue indicates that expression of
Cathepsin K mRNA increases in root-resorping tissue compared with
the normal peridontum. It is believed that ondotoclast in the
root-resorping tissue expressed Cathepsin K mRNA that participates
in proteolysis during root resorption. Evidence suggests that the
cellular mechanisms of physiological root resorption appear to be
quite similar to those of osteoclastic bone resorption.
[0094] The distribution of Cathepsin K in ondotoblasts has been
found to be similar to that previously seen in osteoclasts.
Furthermore, Cathepsin K-positive fibril-like structures were found
in the vacuoles of fibroblasts. Cathepsin K is suggested to take a
part in the degradation of the dentin matrix (type I collagen
fibrils and non-collagenous protein) of the tooth root, and in
subsequent intracellular degradation of endocytosed fragmented
fibril-like structure in endolysosomes.
Surgical Implant Loosening
[0095] Prosthetic wear debris-induced peri-implant osteolysis is a
major cause of aseptic loosening after total joint replacement.
Cathepsin K and tartrate-resistant acid phosphatase were highly
expressed in both mononucleated and multinucleated cells associated
with the bone surface. Findings suggest that cells expressing the
full repertoire of osteoclast phenotypic markers are involved in
the pathogenesis of peri-implant osteolysis after total joint
replacement. In one study it was found that interface tissue
between the bone and loosening total hip implant is acidic and
highly osteolytic. It is characterized by the formation of
Cathepsin K positive foreign body giant cells. The multinuclear
cells induced with pseudosynovial fluid contained active cathepsin
K protein and were capable of bone matrix resorption in vitro. The
cells were shown to express osteoclast phenotype markers, such as
mRNA for cathepsin K, TRAP and calcitonin receptor.
Infection-Associated Bone Resorption
[0096] Persistent infection can participate in delayed or
non-unions of long bone fractions. In the context of bacterial
infection the generation of osteoclasts may be enhanced. Genes
specific for osteoclasts such as tartrate resistant acid
phosphatase (TRAP) and Cathepsin K are found to be upregulated in
cells of a promonocytic cell line, U937 which were differentiated
to monocytes. Local bacterial infections with shedding of bacterial
lipopolysaccharide may create a micro-environment promoting the
generation of bone resorpting cells. This in turn could contribute
to the infection-associated delayed healing or non-unions of
fractures. The present invention provides a method for treating and
promoting healing in delayed or non-unions of long bone
fractions.
[0097] Delay or non-union of bones can also occur for example after
trauma and without the presence of an infectious element.
Pulmonary Conditions
[0098] Inflammation of airways and lung is often associated with
swelling of airway and lung tissue and infiltration of airway and
lung tissue with leucocytes including neutrophils, eosinophils or
basophils and mast cells. This is well established and seen in
extraordinary common conditions such as common cold, airway
infection, and pulmonary infections, but also in conditions
associated with allergy including allergic rhinitis, asthma or
other allergic conditions. Moreover, this is also seen in acute
diseases associated with inflammation of lung and airways as well
as in chronic obstructive pulmonary disease including chronic
bronchitis, adult chronic bronchitis and emphysema. The course of
COPD is characterized by intermittent exacerbations of the disease.
In an exacerbation, there is also a significant influx of
eosinophils into the tissue contributing to the inflammation.
Lung Fibrosis
[0099] Lung fibrosis is characterized by tissue remodelling
resulting from an imbalance between synthesis and degradation of
extracellular organic matrices. In lung tissue cathepsin K
transcription is the most strongly upregulated in response to
silica, and this upregulation is related to the fibrotic process.
Pulmonary macrophages and fibroblasts have been identified as
cathepsin K overproducing cells in the lung of silicotic mice.
Interstitial fibrosis of the lung is a final common pathway of many
different lung diseases, such as idiopathic interstitial
pneumonitis (idiopathic pulmonary fibrosis) and granulomatous
diseases (sarcoidosis). Fibrotic areas are also commonly found in
patients suffering from chronic obstructive pulmonary disease.
There is evidence to support that cathepsin K has a specific role
in the development of lung fibrosis. Higher cathepsin K-mediated
collagenolytic activity in fibroblasts of fibrotic lungs indicates
that cathepsin K is functionally active and is crucial to degrade
ECM proteins. Modulating the expression and activity of cathepsin
provides a new approach for the treatment and/or prevention of lung
fibrosis.
Chronic Obstructive Pulmonary Disease/Emphysema
[0100] Recent studies suggest that in the setting of inflammation,
such as induced by cigarette smoking, cells that do not normally
express cathepsin K begin to do so and in doing so may act like
osteoclasts. Studies in human vascular smooth muscle cells
exemplify this point. Vascular smooth muscle cells do not normally
express with cathepsin S or K in the setting of evolving
atherosclerosis, however, there is marked upregulation of both
enzymes in medial and nonintimal smooth muscle cells.
Obesity
[0101] In obesity, adipocytes undergo dramatic morphological and
molecular changes associated with alterations in their gene
expression profile. Studies of white adipose tissue of obese
selected mice indicates that white adipose tissue Cathepsin K mRNA
was elevated 5.9-fold as were Mitf and Tfe3 (2- and 3.3-fold
respectively), two transcription factors involved in Cathepsin K
induction in osteoclasts. Moreover, the level of white adipose
tissue Cathepsin K mRNA was increased in other obese models of mice
and decreased in mice undergoing weight loss. Cathepsin K is a
novel marker of obesity and can be considered a target for
inhibition of adipose mass growth.
Experimental Studies
[0102] The inventors have investigated the potential of an
oversulfated polysaccharide namely, calcium XPS, to act as an
inhibitor of cathepsin K collagenolytic and bone resorptive
activity. In particular the inventors studied the effect of an
inhibitor on osteoclast formation and function.
EXAMPLE 1
[0103] (i) Insoluble Bone Collagen
[0104] Fresh ovine bone was milled in a stainless steel mortar and
pestle in liquid nitrogen. The resultant bone powder was defatted,
decalcified and freeze-dried before storage at 4.degree. C.
(ii) Cathepsin K Bone Collagen Digests [0105] 2 mg of insoluble
bone collagen powder was digested in 100 mM sodium acetate pH 5.5
containing 2.5 mM dithiothreitol and 2.5 mM EDTA with 190 mM
Cathepsin K and increasing doses of PPS. [0106] Digestions were
incubated at 37.degree. C. for 24 hours and terminated by the
addition of 2 .mu.M E.sub.84. Residue and supernatant were
hydrolysed separately and the hydroxyproline content measured.
(iii) RAW 264.7 Cells [0107] Murine macrophage RAW 264.7 cells
respond to Receptor Activator NF.sub.KB Ligand (RANKL) stimulation
to generate bone pit resorptive osteoclasts. [0108] RAW cells were
pre-cultured for several days in High-Glucose Dulbecco's Modified
Eagles Medium (HG-DMEM) with 10% Fetal Calf Serum (FCS). [0109]
Cells were seeded at 250,000 cells/mL in HG-DMEM/10% FCS
with/without 20 mg/mL RANKL and increasing doses of PPS (0-50
.mu.g/mL). [0110] Cells were incubated at 37.degree. C., 5%
CO.sub.2 for 4 days. [0111] The number of multinucleated cells
present in each was counted by two observers at day 2, 3 and 4. The
cells were then harvested for RNA extraction and subsequent RT-PCR.
(iv) H&E Staining [0112] Identical cultures were prepared in
petri dishes containing pre-labelled slides and incubated for 4
days, after which slides were carefully rinsed in PBS, immersed in
ice-cold acetone and air-dried before freezing. [0113] The slides
were then stained with H&E before multinucleated osteoclasts
were photographed and counted. (v) Results
[0114] The addition of RANKL (20 ng/mL) to RAW cells resulted in
significant development of multinucleated cells by day 3 (FIGS. 2
and 3; p<0.05), confirming RANKL is able to differentiate RAW
cells into OCL cells.
[0115] FIG. 4A shows that the number of cells containing 2-3 nuclei
as a percentage total cells was still significantly increased with
the addition of RANKL at most concentrations of CaXPS. At 50
.mu.g/mL CaXPS, there were significantly more 2-3 nucleated cells
in the presence of RANKL (p<0.05).
[0116] In contrast, RANKL-induced formation of true multinucleated
(>4 nuclei/cell) OCL cells was inhibited by 10 and 50 .mu.g/mL
CaXPS (FIG. 4B). Interestingly, at 0.5 .mu.g/mL CaXPS, there was
significantly more RANKL-induced multinucleated OCL cell formation
than in the absence of CaXPS (Fib. 4B; p<0.05).
[0117] The expression of Cathepsin K (FIG. 5A) and MMP-9 (FIG. 5B)
by RAW cells was increased by RANKL, which is consistent with the
formation of OCL cells. Increasing concentrations of CaXPS in the
absence of RANKL had no marked effect on either Cathepsin K or
MMP-9 gene expression. In the presence of RANKL, a significant
increase was seen in Cathepsin K expression at the lowest dose of
CaXPS whereas a decrease in MMP-9 expression was seen at 5 .mu.g/mL
CaXPS (p<0.05).
[0118] Taken together, these results suggest the use of CaXPS in
the treatment of disease through its down regulation of osteoclast
formation and Cathepsin K activity. Ongoing studies will determine
the effect of CaXPS on bone resorption by osteoclasts in vitro and
in vivo.
* * * * *