U.S. patent application number 12/302511 was filed with the patent office on 2009-08-06 for sulphated hyaluronic acid for treating degenerative osteoarthritis.
Invention is credited to Lanfranco Callegaro, Davide Renier.
Application Number | 20090197807 12/302511 |
Document ID | / |
Family ID | 38353080 |
Filed Date | 2009-08-06 |
United States Patent
Application |
20090197807 |
Kind Code |
A1 |
Callegaro; Lanfranco ; et
al. |
August 6, 2009 |
SULPHATED HYALURONIC ACID FOR TREATING DEGENERATIVE
OSTEOARTHRITIS
Abstract
This invention relates to oral and intra-articular formulations
based on sulphated hyaluronic acid which are effective in the
treatment of degenerative osteoarthritis.
Inventors: |
Callegaro; Lanfranco; (Abano
Terme, IT) ; Renier; Davide; (Abano Terme,
IT) |
Correspondence
Address: |
MATHEWS, SHEPHERD, MCKAY, & BRUNEAU, P.A.
29 THANET ROAD, SUITE 201
PRINCETON
NJ
08540
US
|
Family ID: |
38353080 |
Appl. No.: |
12/302511 |
Filed: |
May 3, 2007 |
PCT Filed: |
May 3, 2007 |
PCT NO: |
PCT/EP2007/003920 |
371 Date: |
November 26, 2008 |
Current U.S.
Class: |
514/1.1 ;
514/54 |
Current CPC
Class: |
A61K 9/1652 20130101;
A61K 9/0019 20130101; A61P 19/08 20180101; A61K 45/06 20130101;
A61K 38/014 20130101; A61K 38/39 20130101; A61K 9/4866 20130101;
A61P 19/00 20180101; A61K 31/726 20130101; A61K 31/737 20130101;
A61P 19/02 20180101; A61K 31/737 20130101; A61K 2300/00 20130101;
A61K 31/726 20130101; A61K 2300/00 20130101; A61K 38/39 20130101;
A61K 2300/00 20130101; A61K 38/014 20130101; A61K 2300/00
20130101 |
Class at
Publication: |
514/12 ;
514/54 |
International
Class: |
A61K 31/728 20060101
A61K031/728; A61P 19/00 20060101 A61P019/00; A61K 8/65 20060101
A61K008/65 |
Foreign Application Data
Date |
Code |
Application Number |
May 31, 2006 |
IT |
PD2006 A 000219 |
Claims
1. Pharmaceutical composition containing sulphated hyaluronic acid
for the prevention and/or treatment of cartilage damage caused by
degenerative osteoarthritis and/or wear due to aging of the joint
structure.
2. Pharmaceutical composition as claimed in claim 1, wherein the
molecular weight of the hyaluronic acid is between 50.000 and
800.000 D.
3. Pharmaceutical composition as claimed in claim 2, wherein the
molecular weight of the hyaluronic acid is between 100.000 and
230.000 D.
4. Pharmaceutical composition as claimed in claim 2, wherein the
molecular weight of the hyaluronic acid is between 500.000 and
750.000 D.
5. Pharmaceutical composition as claimed in claim 1, wherein the
degree of sulphation of the sulphated hyaluronic acid is between
0.5 and 3.5.
6. Pharmaceutical composition as claimed in claim 5, wherein the
degree of sulphation of the sulphated hyaluronic acid is between
0.5 and 1.5.
7. Pharmaceutical composition as claimed in claim 1, wherein the
hyaluronic acid is produced by extraction, fermentation or
technological means.
8. Pharmaceutical composition as claimed in claim 1, containing
collagen and/or hydrolysed collagen and/or glycosaminoglycans.
9. Pharmaceutical composition as claimed in claim 1, containing
pharmacologically and/or biologically active substances.
10. Pharmaceutical composition as claimed in claim 9, wherein the
pharmacologically and/or biologically active substances are
selected from the group of vitamins and/or mineral salts and/or
hormones and/or antibiotics and/or growth factors.
11. Pharmaceutical composition as claimed in claim 1, for the
prevention and/or oral treatment of cartilage damage caused by
degenerative osteoarthritis and/or wear due to aging of the joint
structure.
12. Pharmaceutical composition as claimed in claim 11, wherein the
concentration of sulphated hyaluronic acid is between 75 and 400 mg
per dose unit.
13. Pharmaceutical composition as claimed in claim 1, for the
intra-articular treatment of cartilage damage caused by
degenerative osteoarthritis and/or wear due to aging of the joint
structure.
14. Pharmaceutical composition as claimed in claim 13, wherein the
concentration of sulphated hyaluronic acid is between 1 and 100
mg/ml.
15. Pharmaceutical composition as claimed in claim 14, wherein the
concentration of sulphated hyaluronic acid is between 10 and 20
mg/ml.
16. Use of sulphated hyaluronic acid as claimed in claim 1 for the
preparation of formulations suitable for the prevention and/or oral
treatment of cartilage damage caused by degenerative osteoarthritis
and/or wear due to aging of the joint structure.
17. Use of sulphated hyaluronic acid as claimed in claim 1 to
prepare injectable formulations suitable for the intra-articular
treatment of cartilage damage caused by degenerative osteoarthritis
and/or wear due to aging of the joint structure.
Description
[0001] The present invention relates to oral and intra-articular
formulations based on sulphated hyaluronic acid which are effective
in the treatment of degenerative osteoarthritis.
FIELD OF INVENTION
[0002] Osteoarthritis (OA) is a very common disorder, characterised
by progressive degeneration of the joint cartilage which thins and
deteriorates following loss of the cellular and extracellular
component, and can even disappear entirely. The causes of the
disease are only partly clear. They basically seem to involve a
series of processes triggered by a mechanical imbalance that
affects the entire joint, for example after a trauma or mechanical
stress. Inside the cartilage there is a delicate balance between
destruction of the exhausted extracellular matrix and formation of
intact matrix. All this depends largely on enzymes produced by the
chondrocytes, a cellular component of joint matrix. In the event of
an excessive or unbalanced load on the joint, an inflammatory
situation arises. This triggers the release of inflammatory
cytokines which, in turn, stimulate the chondrocytes to produce
metalloproteases (MMP), enzymes responsible for cartilage
destruction, thus altering the balance between synthesis and
degradation of the extracellular matrix. The scientific literature
also describes in detail the cell death processes which affect the
chondrocytes when they come into contact with substances produced
by the breakdown of the matrix (Cao et al., Exp Cell Res, 1999,
246:527-37).
[0003] In view of the factors described above, degenerative
osteoarthritis can be defined as a chronic disease which normally
occurs after an initial acute stage of inflammation, known as
inflammatory osteoarthritis. The presence or absence of an
inflammatory state in the joint (caused by cytokines and MMPs
together with other substances) distinguishes the acute
inflammatory stage of the disease (inflammatory osteoarthritis)
from the chronic non-inflammatory stage (degenerative
osteoarthritis). Degenerative osteoarthritis does not only occur as
the chronic stage of inflammatory osteoarthritis, but also results
from physiological aging of the joint cartilage.
[0004] The integrity of the extracellular matrix is therefore
crucial to the survival of the chondrocytes, and consequently
essential for healthy cartilage.
[0005] The cartilage matrix is a three-dimensional structure
consisting of collagen molecules and aggregated complexes of
proteoglycans, which in turn are formed by [0006] a skeleton of
hyaluronic acid (HA), [0007] glycosaminoglycans (GAG) containing
repetitive disaccharide units of glucosamine or galactosamine,
which in turn carry negatively-charged carboxyls or sulphate
groups, as a result of which the GAGs form long negatively-charged
chains, [0008] polypeptides.
[0009] HA is a polysaccharide molecule with considerable
viscoelastic properties. It is present in the joint cavities as a
fundamental component of synovial fluid, where it acts as a
lubricant and shock-absorbing agent, and protects the chondrocytes
against the action of the inflammatory cytokines (Asari et al.,
Arch Histol Cytol, 1995, 58:65-76; Brun et al., Osteoarthr Cartil,
2003, 11:208-16; Stove et al., J Orthop Res, 2002, 20:551-5). HA,
as such or in derivatised form, has long been used to treat
degenerative osteoarthritis, either as a "viscosupplement" or a
lubricant.
[0010] The various products include (by way of example)
Hyalgan.RTM. (HA purified from rooster combs according to EP 138572
B1), Synvisc.RTM. (Hylan G-F20, namely HA crosslinked with
formaldehyde and divinyl sulphone, as described in U.S. Pat. No.
4,713,448), and Artz.RTM. (HA with a MW of between 620 and 1200
KDa), which can only be administered by the intra-articular
route.
[0011] The oral treatments according to the prior art involve the
administration of sulphated GAGs such as glucosamine sulphate and
chondroitin sulphate in association with HA and hydrolysed
collagen, in order to promote HA synthesis in the joint cavity,
reduce inflammation, and protect the chondrocytes in an
osteoarthritic joint (U.S. Pat. No. 6,645,948; U.S. Pat. No.
6,476,005).
[0012] Scientific evidence clearly demonstrates the efficacy of
sulphated GAGs in increasing the HA content in the synovial fluid
of patients suffering from inflammatory osteoarthritis (McCarty et
al., Med Hypoth, 2000, 54:798-802; McCarty M F, Med Hypoth, 1998,
50:507-510).
[0013] Further patents and patent applications are also known which
describe new treatments for inflammatory osteoarthritis involving
intra-articular administration of HA in association with
chondroitin sulphate (U.S. Pat. No. 6,906,044), or claiming oral
treatment with HA at given doses (U.S. Pat. No. 6,607,745),
possibly in association with glucosamine, chondroitin sulphate or
glucosamine sulphate (U.S. Pat. No. 6,924,273); finally, the use of
HA with high molecular weight for the prevention and treatment of
osteoporosis is known (patent application WO2005/032276).
[0014] In addition to glucosamine sulphate and chondroitin
sulphate, another sulphated GAG used for chondroprotection is HA,
suitably sulphated as described in the prior art, to which
anti-inflammatory, anticoagulant and cell-adhesion-inhibiting
effects are attributed.
[0015] In particular, US 2004 0,053,885 discloses the use of
sulphated hyaluronic acid (subsequently referred to as HA-S) in the
intra-articular treatment of inflammatory arthritis, and
specifically rheumatoid arthritis (autoimmune disease), while EP
754460 B1 claims the use of HA-S in the parenteral injectable
treatment of inflammatory states, including joint rheumatism.
[0016] The present invention supersedes the prior art because it
relates to formulations based on chemically sulphated HA, for oral
administration or intra-articular injection, which effectively slow
joint degeneration and promote reconstruction of the extracellular
matrix in joints affected by chronic degenerative osteoarthritis,
which is consequently not at the inflammatory stage (these
compositions therefore cannot be used to treat inflammatory
osteoarthritis). According to a particularly preferred aspect, the
present invention relates to formulations based on HA chemically
modified by sulphation as described in EP 702699 B1: the Applicant
has demonstrated the therapeutic efficacy of HA-S by comparison
with a standard treatment based on sulphated glycosaminoglycans in
the treatment of osteoarthritic cartilage deficiency, and compared
its efficacy with non-sulphated HA to demonstrate the clear
pharmacological superiority of HA-S over both GAGs and HA.
DETAILED DESCRIPTION OF THE INVENTION
[0017] The present invention relates to formulations based on HA
chemically modified by sulphation, which are effective in the
(preferably oral) treatment of chronic forms of degenerative
osteoarthritis without inflammatory sequelae.
[0018] As already stated, the process of degenerative
osteoarthritis causes progressive damage to the extracellular
matrix in the joint cartilage. This damage is manifested by
thinning of the matrix and loss of the chondrocyte component; the
process culminates with the total destruction of the joint
cartilage. The disorder can be manifested as chronicisation of
inflammatory osteoarthritis or represent the physiological
development of the joint aging process.
[0019] From the morphological standpoint the cartilage can be
divided into four different zones, from the joint surface to the
subchondral bone, namely: [0020] zone I: the thinnest, outermost
part, which is only partly cellularised, [0021] zone II: the
transitional zone, which contains the highest percentage of cells
and is basically responsible for matrix production, [0022] zone
III: the radial zone, which is the largest; the entire thickness of
the cartilage depends on its structure, [0023] zone IV: the
calcified zone, a thin layer that separates the radial zone from
the subchondral bone.
[0024] The deeper the zone affected, the more serious is the damage
caused by degenerative osteoarthritis. It develops with the same
processes, involving the same zones, whether it is due to
inflammatory disease or results from physiological joint aging
caused by advancing age or overload.
[0025] A pharmacological treatment able to provide the
extracellular matrix with the "building blocks" it needs to
reconstitute its complex structure of proteoglycans, in order to
strengthen its load-bearing skeleton and prevent cell loss, would
be most useful at the stage of chronicisation of the disorder. This
invention demonstrates that the administration of sulphated
hyaluronic acid represents a valid therapeutic approach for these
purposes because it increases, within the joint, the presence of
the material that constitutes the "scaffolding" of the cartilage
matrix and, by ensuring its structural integrity, prevents its
decellularisation. The formulations of the invention can therefore
also be used for the prevention of cartilage damage.
[0026] The in vivo trial performed by the Applicant demonstrates
that HA-S slows degradation and stimulates regeneration of the
basic structure of the extracellular matrix to a surprisingly
greater extent than other GAGs and non-sulphated HA.
[0027] The sulphated hyaluronic acid suitable for the purpose of
this invention is prepared according to a process described in EP
702699 B1: sulphation is performed with the SO.sub.3-pyridine
complex, and involves the alcoholic hydroxyls present in the
polysaccharide chain.
[0028] The degree of sulphation can range between 0.5 and 3.5 (EP
0940410 B1), and is preferably between 0.5 and 1.5 (the average
grade is defined as 1:1 sulphate group to disaccharide group),
starting from HA with different molecular weights, ranging from
50.000 to 800.000 D, preferably 100.000 to 230.000 D, using HA
produced by extraction, fermentation or technological means.
[0029] The derivative obtained maintains all the physical
characteristics of the starting polymer; in particular, the
molecular weight of HA is not changed by the sulphation process,
consequently allowing the same viscosity as the starting
polysaccharide to be maintained.
[0030] The experiments described below were conducted by comparing
HA-S produced by fermentation, having an average molecular weight
of 180/200 KD and an average degree of sulphation of 1, with a
reference treatment based on Condral.RTM.
(galactosaminoglucuronoglycan sulphate, the depolymerised form of
chondroitin sulphate) and with a treatment based on HA (with a
molecular weight of 180/200 KD) against the untreated control.
[0031] Trial Design: Inducement and Treatment of Degenerative
Osteoarthritis
[0032] The experiments were conducted on adult rabbits, in strict
compliance with current legislation. Degenerative osteoarthritis of
the right knee was induced unilaterally by surgical resection of
the anterior cruciate ligament according to a well-established
experimental model (Yoshimi et al., Clin Orthop Relat Res, 1994,
298:296-304). The experimental model used does not trigger an
inflammatory process, as demonstrated by the total absence of
plasma infiltrates in the joint cavities of all the treated
animals, including the controls (Table). Surgical ablation of the
ligament consequently represents a good example of experimental
degenerative osteoarthritis, which allows correct pharmacological
evaluation of the formulations tested.
[0033] The animals were divided into 4 homogenous groups (6 animals
per group) and treated orally, 24 hours after surgery, every day
for 61 days, with: [0034] saline solution 0.9% NaCl (Control);
[0035] HA-S at the dose of 5.5 mg/kg; [0036] HA at the dose of 20
mg/kg; [0037] Condral.RTM. at the dose of 20 mg/kg.
[0038] At the end of the treatment the animals were killed and
examined macroscopically to check for the absence of systemic
toxicity. Next, the lateral and medial condyles of the femur with
the tibial plate were fixed in formalin buffered to 10% and
embedded in paraffin wax. The preparations were then processed, by
a method known to one skilled in the art, to obtain sections
analysable under the optical microscope after staining with
haematoxylin-eosin.
[0039] The cartilage tissue removed from the medial and lateral
femoral condyles was then subjected to semiquantitative
histopathological analysis and histomorphometric analysis. In
particular, parameters relating to degeneration, the structural
integrity of the cartilage and the extent of abrasion on the
thickness of the cartilage were taken into consideration.
[0040] Analysis of Results
TABLE-US-00001 TABLE Synovial membrane: Cartilage tissue: Group
plasma infiltrates plasma infiltrates Control 0 0 HA-S 0 0 HA 0 0
Condral 0 0
[0041] As already stated, the total lack of plasma infiltrates in
the synovial fluid and the cartilage confirms the complete absence
of inflammation in the joint cavity in the controls treated with
NaCl and in all the groups studied.
[0042] Macroscopic analysis of the animals and their organs also
demonstrated the lack of systemic toxicity of the pharmacological
compositions tested.
[0043] FIG. 1 summarises the data collected from the cartilage
examination: in the group of animals treated with saline solution
(controls), serious degeneration characterised by morphological
alteration of the cell component was evident. The situation of the
animals treated with Condral.RTM. was better; in this case the
cartilage degeneration was estimated at 2.8, and was therefore
moderate/marked; it is important to note that this result is very
similar to that obtained with HA treatment. The cartilage of the
animals treated with HA-S was in a much better situation: the level
of degradation was slight (1,7). Oral administration of HA-S
therefore proved effective in significantly slowing the process of
degradation of the cartilage of joints affected by degenerative
osteoarthritis.
[0044] A second, important evaluation relates to analysis of the
structural integrity of the cartilage tissue, which is measured,
according to recognised standards, in terms of irregularity of the
surface and of the cartilage structure (FIG. 2). This graph, like
the preceding one, confirms that treatment with HA-S helps to
maintain good structural integrity, to a significant extent
compared with the controls, and also compared with Condral.RTM.
treatment. The group of animals treated with HA-S only presented
slight cracks in the transitional cartilage zone, whereas those
treated with Condral.RTM. presented marked lesions of the radial
zone; the finding relating to treatment with HA, which again proved
as effective as Condral.RTM., is particularly interesting.
[0045] Finally, FIG. 3 illustrates the depth of the lesion along
the entire thickness of the cartilage, the maximum lesion being one
which causes exposure of bone tissue following total loss of
cartilage.
[0046] The data illustrate the mean values of the lesions of the
lateral and medial condyles; once again it was confirmed that oral
treatment with HA-S is more effective than treatment with
Condral.RTM. or treatment with HA alone in preserving the integrity
of the cartilage matrix.
[0047] Analysis of the data presented here clearly indicates that
HA-S is more effective than sulphated GAGs and HA in the treatment
of non-inflammatory osteoarthritis, whether it results from
trauma/disease or the normal joint aging process.
[0048] If necessary, HA-S could possibly be associated with other
molecules useful to strengthen the extracellular matrix, such as
collagen (possibly hydrolysed) or other GAGs, or with other
pharmacologically and/or biologically active substances such as
growth factors and/or hormones, vitamins (especially vitamins A, C,
D, and E, and group B in general), antibiotics and mineral salts
(especially calcium, magnesium and selenium salts and other trace
elements). The procedures described and claimed herein therefore
represent a definite improvement on the current oral treatments for
the disorder in question.
[0049] The surprising results obtained with oral administration of
HA-S, and the well-known chemico-physical and pharmacological
characteristics of hyaluronic acid derivatives, also suggest the
possible administration of these formulations by injection, in
particular intra-articular injection, limited to forms of chronic
degenerative osteoarthritis in which there are no inflammatory
symptoms. In this case too, HA-S could possibly be associated with
other molecules such as other GAGs or other pharmacologically
and/or biologically active substances such as growth factors and/or
hormones, vitamins, antibiotics and antivirals.
[0050] Some examples of preparation of the formulations containing
HA-S of the invention, for oral and intra-articular administration,
are described below by way of example and not of limitation. HA-S
can be formulated in all known ways according to the state of the
art in association with stabilisers, excipients, preservatives
and/or other substance considered useful to obtain the best
possible formulation, for the preparation of granulates,
suspensions, solutions, capsules and tablets.
[0051] Oral Formulations
[0052] The HA used in these formulations has an average degree of
sulphation of 1, and an average molecular weight of 180/200 KD; the
concentration of the Active ingredient ranges between 50 and 400 mg
per dose unit.
Example 1
HA-S 75 mg Rigid Gelatin Capsule
TABLE-US-00002 [0053] Constituent Amount (mg/cap) Active
ingredient: HA-S 75 Excipients Lactose q.s. for the capacity of the
00 capsule Corn starch 16.5 Anhydrous colloidal silicon dioxide 7.7
Magnesium stearate 3.3
[0054] Initially mix the HA-S, lactose, corn starch and anhydrous
colloidal silicon dioxide. Add magnesium stearate to the premix
obtained, and mix. Distribute the final mixture between rigid
gelatin capsules format 00.
Example 2
HA-S 200 mg Rigid Gelatin Capsule
TABLE-US-00003 [0055] Constituent Amount (mg/cap) Active
ingredient: HA-S 200 Excipients Microcrystalline cellulose q.s. for
the capacity of the 00 capsule Corn starch 35.1 Anhydrous colloidal
silicon dioxide 5 Magnesium stearate 5
[0056] Initially mix the HA-S, microcrystalline cellulose, corn
starch and anhydrous colloidal silicon dioxide. Add magnesium
stearate to the premix obtained, and mix. Distribute the final
mixture between rigid gelatin capsules format 00.
Example 3
HA-S 400 mg Rigid Gelatin Capsule
TABLE-US-00004 [0057] Constituent Amount (mg/cap) Active
ingredient: HA-S 400 Excipients Microcrystalline cellulose q.s. for
the capacity of the 00 capsule Corn starch 35.1 Anhydrous colloidal
silicon dioxide 5 Magnesium stearate 5
[0058] Initially mix the HA-S, microcrystalline cellulose, corn
starch and anhydrous colloidal silicon dioxide. Add magnesium
stearate to the premix obtained, and mix. Distribute the final
mixture between rigid gelatin capsules format 00.
Example 4
HA-S 400 mg Granulate
TABLE-US-00005 [0059] Constituent Amount (mg/sachet) Active
ingredient: HA-S 400 Excipients Microcrystalline cellulose 65 Corn
starch 15.5 Sorbitol 2250 Polyvinylpyrrolidone 47.5 Citric acid 10
Aspartame 32 Orange flavouring 180
[0060] Initially mix the HAS1, microcrystalline cellulose and corn
starch. Mix with the binder solution consisting of water and
polyvinylpyrrolidone. When the mixture is uniformly moist,
granulate through a 2 mm sieve. Dry the granulate, then sieve the
dried granulate, forcing it through an 0.8 mm sieve. Mix the
granulate powder with sorbitol, citric acid, aspartame and orange
flavouring. Fill sachets with the granulate.
[0061] Injectable Formulations
[0062] HA-S grade 1 or 2, having an average molecular weight of
180/200 KD or 500/750 KD, prepared in sterile, pyrogen-free saline
solution at the concentration of 1-100 mg/ml, preferably 5-50
mg/ml, and even more preferably 10-20 mg/ml, is preferably used for
the injectable intra-articular preparations. The final solution
must be sterile and pyrogen-free. It can also be freeze-dried and
reconstituted at the time of use.
* * * * *