U.S. patent application number 14/481135 was filed with the patent office on 2014-12-25 for chitosan beads and filler comprising such beads.
The applicant listed for this patent is MERZ PHARMA GmbH & CO., KGaA. Invention is credited to Peter Boderke, Bernhard Hauptmeier, Kevin Kiehm.
Application Number | 20140377368 14/481135 |
Document ID | / |
Family ID | 42112242 |
Filed Date | 2014-12-25 |
United States Patent
Application |
20140377368 |
Kind Code |
A1 |
Kiehm; Kevin ; et
al. |
December 25, 2014 |
CHITOSAN BEADS AND FILLER COMPRISING SUCH BEADS
Abstract
The present invention pertains to chitosan beads consisting of
chitosan cross-linked with citrate ions. The present invention
furthermore pertains to a filler comprising such chitosan-citrate
beads. In one embodiment of the instant invention the filler is a
dermal filler. In one further embodiment of the present invention
the dermal filler is for the treatment of wrinkles and/or folds. In
another embodiment of the instant invention the filler is for use
in the treatment of a medical condition. The filler provided in the
present invention may further comprise one or more active
pharmaceutical ingredients. Further, the present invention pertains
to a process for preparing the filler as claimed herein.
Inventors: |
Kiehm; Kevin; (Telgte,
DE) ; Hauptmeier; Bernhard; (Gelnhausen, DE) ;
Boderke; Peter; (Schwalbach, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MERZ PHARMA GmbH & CO., KGaA |
Frankfurt am Main |
|
DE |
|
|
Family ID: |
42112242 |
Appl. No.: |
14/481135 |
Filed: |
September 9, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13639004 |
Dec 12, 2012 |
8865879 |
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PCT/EP2011/001736 |
Apr 7, 2011 |
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14481135 |
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61321950 |
Apr 8, 2010 |
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Current U.S.
Class: |
424/499 ;
514/55 |
Current CPC
Class: |
Y10T 428/2982 20150115;
A61K 2800/72 20130101; A61Q 19/08 20130101; A61K 2800/654 20130101;
A61K 2800/54 20130101; A61L 27/20 20130101; A61K 2800/56 20130101;
A61L 27/20 20130101; A61K 8/735 20130101; A61L 31/042 20130101;
A61K 8/736 20130101; A61K 8/025 20130101; A61K 31/167 20130101;
A61K 8/365 20130101; A61K 9/1652 20130101; A61K 8/0204 20130101;
A61K 2800/30 20130101; A61L 31/042 20130101; C08L 5/08 20130101;
C08L 5/08 20130101 |
Class at
Publication: |
424/499 ;
514/55 |
International
Class: |
A61K 8/02 20060101
A61K008/02; A61K 31/167 20060101 A61K031/167; A61Q 19/08 20060101
A61Q019/08; A61K 8/73 20060101 A61K008/73 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 8, 2010 |
EP |
10003795.1 |
Claims
1. A dermal filler comprising chitosan beads and an anesthetic
agent, wherein the chitosan beads consist essentially of chitosan
and citrate ions, wherein the chitosan has an average molecular
weight of from about 50 kD to about 5000 kD and is deacetylated to
a degree of from about 70% to about 100%, wherein the chitosan
beads exhibit a mass median diameter of less than or equal to 1500
.mu.m as determined by microscopical analysis.
2. The dermal filler of claim 1, wherein the mass median diameter
of the chitosan beads remains less than or equal to 1500 .mu.m for
a period of 6 months at 25.degree. C..+-.2.degree. C. and 60%.+-.5%
relative humidity as determined by laser diffraction analysis.
3. The dermal filler of claim 1, wherein the chitosan beads have a
shelf-life of 6 months at 25.degree. C..+-.2.degree. C. and
60%.+-.5% relative humidity.
4. The dermal filler of claim 1, further comprising at least one
polysaccharide.
5. The dermal filler of claim 4, wherein the polysaccharide is
hyaluronic acid or a salt thereof.
6. The dermal filler of claim 1, wherein the anesthetic agent is
selected from procaine benzocaine, chloroprocaine, cocaine,
cyclomethycaine, dimethodcaine, larocaine, propoxycaine,
proparacaine, tretracaine, lidocaine, articaine, bupivacaine,
carticaine, cinchocaine, etidocaine, levobupivacaine, mepivacaine,
peperocaine, prilocaine, ropivacaine, and trimecaine.
7. The dermal filler of claim 1, wherein the anesthetic agent is
present in the dermal filler at a concentration from about 0.01% to
6% weight % (w/w %) based on the total weight of the
composition.
8. The dermal filler of claim 1, further comprising
pharmaceutically acceptable isotonicity agents or
preservatives.
9. The dermal filler of claim 1, further comprising
pharmaceutically acceptable exicipents selected from antioxidants,
viscosity enhancers/modifiers, hydrating agents, bulking
substances, tonicity agents, preservatives and surface active
agents, or a mixture thereof.
10. The dermal filler of claim 1, wherein the chitosan beads are
present in the dermal filler in an amount effective for the
treatment of wrinkles and/or folds.
11. The dermal filler of claim 1, wherein the chitosan beads are
present in the dermal filler in an amount effective for the
treatment of frown lines, glabellar lines, nasal furrows, cheek
wrinkles, crow's feet and perioral wrinkles, nasolabial folds, lip
augmentation, forehead wrinkles, mild to moderate nasal furrows and
acne scars.
12. The dermal filler of claim 1, wherein the chitosan beads are
present in the dermal filler in an amount effective for filling a
lip in lip augmentation.
13. The dermal filler of claim 1, wherein the chitosan beads are
present in the dermal filler at a concentration from about 10%
(v/v) to about 95% (v/v) to the total volume of the filler.
14. The dermal filler of claim 1, wherein the chitosan beads are
present in the filler at a concentration from about 25% (v/v) to
about 95% (v/v) to the total volume of the filler.
15. The dermal filler of claim 1, wherein the chitosan beads are
present in the filler at a concentration from about 50% (v/v) to
about 95% (v/v) to the total volume of the filler.
16. The dermal filler of claim 1, wherein the chitosan beads are
present in the filler at a concentration from about 75% (v/v) to
about 95% (v/v) to the total volume of the filler.
17. The dermal filler of claim 1, wherein the chitosan beads are
present in the filler at a concentration from about 85% (v/v) to
about 90% (v/v) to the total volume of the filler.
18. The dermal filler of claim 1, wherein the dermal filler has a
shelf-life of 6 months at 25.degree. C..+-.2.degree. C. and
60%.+-.5% relative humidity.
Description
FIELD OF THE INVENTION
[0001] The present invention pertains to chitosan beads consisting
of chitosan cross-linked with citrate ions. The present invention
furthermore pertains to a filler comprising such chitosan-citrate
beads. In one embodiment of the instant invention the filler is a
dermal filler. In one further embodiment of the present invention
the dermal filler is for the treatment of wrinkles and/or folds. In
another embodiment of the instant invention the filler is for use
in the treatment of a medical condition. The filler provided in the
present invention may further comprise one or more active
pharmaceutical ingredients. Further, the present invention pertains
to a process for preparing the filler as claimed herein.
BACKGROUND OF THE INVENTION
[0002] Treatment with fillers is known since 1980s. Today's most
preferred fillers can be classified as hyaluronic acid-based
fillers (Hylaform.RTM., Hylaform.RTM. Plus, Restylane.RTM.,
Perlane.RTM., Juvederm.RTM., Juvederm.RTM. Ultra, Juvederm.RTM.
Ultra Plus, Puragen.RTM., Puragen.RTM. Plus, Matridur.RTM.),
collagen based fillers (Zyderm.RTM. I, Zyderm.RTM. II,
Zyplast.RTM., Atelocollagen.RTM., CosmoDerm.RTM. I, CosmoDerm.RTM.
II, Resoplast.RTM.) and alginate based fillers (e.g. Novabel.RTM.)
as described in DE 10 2004 019 241.
[0003] Collagen is a natural protein of connective tissue. However,
some people suffer from allergic reactions to collagen and thus, an
allergy test is always suggested by the practitioner prior to
injection of fillers comprising collagen. Hyaluronic acid is a
polysaccharide and is naturally found in many tissues of the body.
The unfavorable effect of fillers comprising hyaluronic acid is the
short-lasting result and the need for multiple injections for an
observable effect. Thereby swellings can occur, which decay in 1-3
days. Thus, treatments with collagen and hyaluronic acid based
fillers are costly and painful due to the prerequisite of multiple
injections and allergy tests. Further reported complications for
the fillers is poor syringeability due to high viscosity,
aggregation of the particles in the packaging and non-homogeneous
distribution of the particles at the injection site.
[0004] The use of alginate as filler is known from DE 10 2004 019
241. However, DE 10 2004 019 241 suggests for the long-lasting
effect of the cross-linked alginate particles the use of barium
(paragraph [0031]).
[0005] Therefore, there remains a continuous need for an improved
dermal filler with an improved safety profile, an improved
tolerability profile, improved application characteristics, and a
long-lasting effect.
[0006] Chitosan and its derivatives are very well known natural
substances and have been employed in the formulation of controlled
release systems, i.e. microcapsules and similar colloidal delivery
systems. This is documented in several publications, e.g. M.
Prabaharan "Review Paper: Chitosan derivatives as promising
materials for controlled drug delivery" Journal of Biomaterials
Applications, 2008; 23; 5 and in O. Gaserod et al. "Microcapsules
of alginate-chitosan--I A quantitative study of the interaction
between alginate and chitosan" Biomaterials 19 (1998), 1815-1825;
and in Cai et al. "Biodegradable chitosan scaffolds containing
microspheres as carriers for controlled transforming growth
factor-.beta.1 delivery for cartilage tissue engineering" Chinese
Medical Journal 2007; 120 (3); 197-203.
[0007] Several methods and reagents have been tested for their
ability to crosslink chitosan and to form microspheres. Reaction of
chitosan with alginate under different conditions resulted in the
formation of microcapsules that had an alginate-chitosan complex
membrane including either an alginate or a chitosan core, depending
on the reaction conditions (Gaserod et al., loc. cit.). Chitosan
microspheres were prepared from chitosan and sodium
tripolyphosphate solution (Cai et al., loc. cit.).
[0008] WO 2008/103594 describes the use of chitosan and its
derivatives as biomaterial for the treatment, repair and/or
enhancement of bodily tissue insufficiencies of the vocal chords,
muscles, ligaments and cartilage. According to the invention the
use of the biomaterial produces a filling effect. Also disclosed
are chitosan or chitosan-derivative gels, which optionally include
chitin microspheres. In a fist step of the chitin microsphere
preparation, chitosan microspheres were obtained by spray-drying a
chitosan solution in acetic acid/ethanol.
[0009] Several publications describe the use of citrate for the
cross-linking of chitosan. However, when citrate was added to an
emulsion of chitosan in acetic acid-containing solution, only
irregular microparticles were formed, and microspheres could only
be obtained when gelatin was added and co-emulsified (Shu, X. Z and
Zhu, K. J., "Chitosan/gelatin microspheres prepared by modified
emulsification and ionotropic gelation", J. Microencapsulation
2001; 18; 237). Alternatively, microspheres were obtained by
dropping a solution containing chitosan and gelatin into a cold oil
in order to obtain coagulation of gelatin, prior to cross-linking
with citrate (Shu, X. Z and Zhu, K. J., "Controlled drug release
properties of ionically cross-linked chitosan beads: the influence
of anion structure", Int. J. Pharm. 2002; 233; 217). Thus, in these
publications the term "chitosan microspheres" denotes microspheres
comprising cross-linked chitosan and gelatin.
OBJECTS OF THE INVENTION
[0010] Accordingly, in view of the problems of the prior art, the
first object of the present invention is to provide
chitosan-citrate beads that are free from gelatine.
[0011] The second object of the present invention is a novel
filler, which is injected below the dermis, thereby leaving no
scar, rapidly restoring volume at the application site and
sustaining the volume augmentation, and which does not contain
collagen, which can cause allergic reactions, thereby not requiring
pre-testing, such as allergic skin testing. It is also important
that the particles remain evenly distributed after the injection to
avoid palpable mass after the carrier is resorbed in the body.
Thus, it is an object of the present invention to provide a novel
filler exhibiting a long-lasting effect and far less side
effects.
[0012] Another object of the present invention is to provide a
novel filler composition, which, unlike conventional fillers, which
contain collagen or hyaluronic acid as a major component, is not
easily degraded by human enzymes or absorbed in the body, thus
ensuring stable longer-lasting volume augmentation, and is cheaper
than conventional fillers.
[0013] One further object of the instant invention is to provide a
filler exhibiting a more improved syringeability as the
conventional fillers, avoidance of aggregation of the particles in
the packaging and non-homogeneous distribution of the particles at
the injection site.
SUMMARY OF THE INVENTION
[0014] These and other objects are solved by chitosan beads, which
consist essentially of chitosan and citrate ions, and a filler
comprising such chitosan beads or chitosan beads comprising
chitosan and citrate ions.
[0015] In one embodiment, the chitosan employed in the chitosan
beads and/or filler according to the instant invention has a
molecular weight distribution from about 50 kD to about 5000
kD.
[0016] In one embodiment of the instant invention, the chitosan
employed is deacetylated to a degree from about 60% to about 100%.
In a particular embodiment, the chitosan employed is deacetylated
to a degree from about 70% to about 90%.
[0017] In one embodiment, the chitosan beads according to the
instant invention, and/or the filler according to the instant
invention comprise chitosan beads that have a mass median diameter
of less than or equal to about 1500 .mu.m determined by
microscopical analysis. In a particular embodiment, the chitosan
beads have a mass median diameter of between about 20 and 1000
.mu.m, more particularly between about 20 and 500 .mu.m, and most
particularly between about 30 and 300 .mu.m.
[0018] In one further embodiment, the present invention pertains to
chitosan beads and/or a filler wherein chitosan beads and/or the
filler may further comprise one or more active pharmaceutical
ingredient selected from the group of anesthetics, analgesics,
anti-microbials, anti-inflammatory drugs, growth factors, hormones,
cosmeceuticals, vitamins, nutrients, stimulants, steroids,
vasoconstrictors, anti-thrombotic agents, anti-coagulation agents,
tranquilizers, muscle relaxants, antifungals, lipolytic agents and
biorejunevation agents.
[0019] In another embodiment of the instant invention, the one or
more active pharmaceutical ingredient may be entrapped, bound to or
encapsulated in the chitosan beads.
[0020] In one further embodiment of the instant invention, the
chitosan beads and/or the filler may further comprise one or more
pharmaceutical excipients selected from antioxidants, viscosity
enhancers/modifiers, hydrating agents, bulking substances, tonicity
agents, preservatives and surface active agents, or a mixture
thereof.
[0021] The chitosan beads, including those of the herein claimed
filler, are stable in form, the form stability being determined
microscopically by recording the changes in spherical shape.
[0022] In one embodiment of the instant invention, the mass median
diameter of the chitosan beads remains constant, i.e. within +/-20%
of the starting value for the mass median diameter, for a period of
at least 6 months, particularly at least 12 months, more
particularly at least 24 months, and most particularly at least 36
months, at 25.degree. C..+-.2.degree. C. and 60%.+-.5% relative
humidity determined by laser diffraction technique.
[0023] In another embodiment of the present invention, the chitosan
beads and/or the filler claimed herein has a shelf-life at
25.degree. C..+-.2.degree. C. and 60%.+-.5% relative humidity of at
least 6 months, particularly at least 12 months, more particularly
at least 24 months, and most particularly at least 36 months.
[0024] In certain embodiments, the chitosan beads, including those
of the filler, according to the instant invention have an
elasticity greater 5%, a tensile strength lower 5 N, and/or a
deformability greater 90%. Deformability and elasticity are
determined according to the method described by Edwards-Levy et.
al. (Biomaterials 20 (1999) 2069-2084) using a texture
analyzer.
[0025] In one embodiment of the instant invention, the chitosan
beads are for use as drug-delivery vehicles.
[0026] In one embodiment of the instant invention, the filler is
for use for aesthetic purposes.
[0027] In one further embodiment of the present invention, the
filler is for use as a dermal filler.
[0028] In one embodiment of the present invention, the dermal
filler is for the treatment of, or for the use in the treatment of,
wrinkles and/or folds.
[0029] In another embodiment of the present invention, the filler
is for the treatment of, or for the use in the treatment of, a
medical condition, including lipoatrophy, gastroesophageal reflux
disease (GERD), urine incontinence, vesico ureteral reflux (VUR),
or a psychological condition caused by the appearance of an
aesthetic deficiency, including, but not limited to, frown lines,
medium depth wrinkles, such as nasolabial folds, lip augmentation,
forehead wrinkles, glabellar lines, obvious mild to moderate nasal
furrows and cheek wrinkles, crow's feet, perioral wrinkles, breast
and acne scars.
[0030] In another embodiment of the present invention, the filler
is used for the treatment of, or for the use in the treatment of,
acne scars, such as by filling areas of acne scars.
[0031] The present invention further pertains to a method of
treating a medical condition, including lipoatrophy,
gastroesophageal reflux disease (GERD), urine incontinence, vesico
ureteral reflux (VUR), or a psychological condition caused by the
appearance of an aesthetic deficiency, including, but not limited
to, frown lines, medium depth wrinkles, such as nasolabial folds,
lip augmentation, forehead wrinkles, glabellar lines, obvious mild
to moderate nasal furrows and cheek wrinkles, crow's feet, perioral
wrinkles and acne scars, wherein said method comprises a step of
administering a filler as claimed in the present invention to a
patient in need thereof
[0032] The present invention further pertains to a method of using
a filler according to the present invention in plastic, cosmetic,
dental or general surgery, in ophthalmology, in orthopedics, as
products for preventing tissue adhesions, or in urology, wherein
said method comprises a step of administering a filler as claimed
in the present invention to a patient in need thereof.
[0033] Further, the present invention pertains to a process (i.e. a
method, such as a manufacturing method) for preparing chitosan
beads and/or the filler as claimed herein, wherein the process
comprises a step of dropping a chitosan solution into an aqueous
solution containing citrate anions.
[0034] In one embodiment in the process for preparing chitosan
beads and/or the filler of the instant invention, the pH of the
aqueous solution containing citrate anions is adjusted to a value
from about 6 to about 11.
[0035] In one further embodiment of the present invention, the
concentration of the citrate ions in the process for preparing said
filler is below about 2.0 M.
[0036] In one embodiment of the present invention, the
concentration of the citrate ions in the process for preparing the
chitosan beads and/or the filler is between about 0.01 M and about
1.0 M.
[0037] In another embodiment of the present invention, the
concentration of chitosan in the process for preparing chitosan
beads and/or the filler is at maximum about 5.0 wt-% (w/w %)
relative to the total weight of the composition, particularly
between about 0.5 wt-% and 4 wt-%, more particularly between about
0.5 wt-% and 3 wt-%, and most particularly between about 0.5 wt-%
and 2 wt-%.
[0038] In one further embodiment of the instant invention, the
viscosity of the chitosan solution in the process for preparing
chitosan beads and/or the filler is in the range from about 50
mPa*s to about 2000 mPa*s measured by a falling ball viscometer at
20.degree. C., particularly between about 100 mPa*s and about 1700
mPa*s, more particularly between about 500 mPa*s and about 1500
mPa*s, and most particularly between about 750 mPa*s and about 1250
mPa*s.
[0039] In one embodiment, the pH of the chitosan solution is
between about pH 1 and about pH 6, more particularly between about
pH 1 and pH 5.
[0040] In another embodiment of the present invention, the aqueous
solution containing citrate ions further comprises one or more
active pharmaceutical ingredients selected from the group of
anesthetics, analgesics, anti-microbials, anti-inflammatory drugs,
growth factors, hormones, cosmeceuticals, vitamins, nutrients,
stimulants, steroids, vasoconstrictors, anti-thrombotic agents,
anti-coagulation agents, tranquilizers, muscle relaxants,
antifungals, lipolytic agents and biorejunevation agents.
[0041] Further, the present invention pertains to a kit comprising
a filler as provided herein and an injection device. The injection
device could be a syringe or an electronic injection device.
[0042] Further, the present invention pertains to an injection
device comprising the filler provided herein. The injection device
could be a prefilled syringe or an electronic injection device.
[0043] In another aspect, the invention relates to a method,
wherein the chitosan beads cross-linked with citrate ions are
redissolved after implantation, to the extent necessary, by the
injection of a solution containing divalent or trivalent
cations.
BRIEF DESCRIPTION OF THE FIGURES
[0044] FIG. 1. Chitosan beads cross-linked with citrate ions and
stored in citrate buffer.
DETAILED DESCRIPTION OF THE INVENTION
[0045] In a first aspect, the present invention relates to chitosan
beads, which consist essentially of chitosan and citrate, i.e.
chitosan cross-linked with citrate ions.
[0046] The term "bead" or "beads" as used in the present invention
relates to spherical particles.
[0047] In the context of the present invention, the term "chitosan
beads, which consist essentially of chitosan and citrate" refers to
beads that are formed from chitosan and citric acid, or a citrate
salt in the presence of an acid, and wherein the resulting chitosan
beads do not contain more than about 5% impurities, such as acid
anions other the citrate, and/or citrate salts, particularly not
more than about 2%, and even more particularly not more than
1%.
[0048] In the context of the present invention, the term
"impurities" includes both (i) impurities present in the starting
materials used for forming the beads, and (ii) any other substances
that may otherwise provide an auxiliary function in the formation
of cross-linked structures, including acid anions other the
citrate, or other polymeric molecules, such as gelatine, or inert
fillers. The term "impurities" does not include, however, any
solvent, solvent mixture or solution, that may be entrapped in the
chitosan beads. Furthermore, the term "impurities" does not include
any active pharmaceutical ingredient or other substance, that is
incorporated in the beads of the present invention, wherein the
beads act as a vehicle for such active pharmaceutical ingredient or
other substance.
[0049] Thus, the present invention relates to chitosan beads,
wherein the three-dimensional network of chitosan cross-linked with
citrate ions forming the bead structure consists essentially of
chitosan and citrate, i.e. wherein that three-dimensional network
of chitosan cross-linked with citrate ions forming the bead
structure does not contain more than 5% impurities, particularly
not more than about 2%, and even more particularly not more than
1%.
[0050] In a second aspect, the present invention relates to a
filler comprising such chitosan beads and/or chitosan beads,
wherein the chitosan beads comprise chitosan and citrate ions.
[0051] The term "filler" as used in the instant invention relates
to compositions which are administered for augmentation, repair or
strengthening of tissue, or for filling a bodily cavity, in a
mammal. The term "mammal" as used herein refers to a human or an
animal taken from the list of farm animals like horses, cattle,
pigs, camels, chicken, turkey, or pets like dogs or cats.
[0052] The term "chitosan" as used in the instant invention relates
to a linear polysaccharide composed of randomly distributed
.beta.-(1-4)-linked D-glucosamine (deacetylated unit) and
N-acetyl-D-glucosamine (acetylated unit) and/or salts thereof.
[0053] In one embodiment, the chitosan is selected from the group
of chitosan glycolate, chitosan lactate, chitosan acetate, chitosan
succinate, N-(aminoalkyl) chitosan, succinyl chitosan,
quateraminated chitosan, octanoyl chitosan, acetyl chitosan, thiol
chitosan, trimethyl chitosan, carboxymethylchitosan,
trimethyammonium chitosane, N-diethyl methyl chitosan, N-methyl
chitosan, carboxymethyl chitosan, N-carboxyethyl chitosan, glycol
chitosan, N-(2-hydroxy)propyl-3-trimethylammonium chitosane or
mixtures thereof.
[0054] In one embodiment the chitosan is produced by deacetylation
of chitin from the exoskeleton of crustaceans and cell walls of
fungi or manufactured by biotechnological and/or enzymatic
methods
[0055] In one embodiment, the chitosan employed is deacetylated to
a degree from about 60% to about 100%. A commercially available
example for such a chitosan is Chitopharm.RTM. from the company
Cognis, such as Chitopharm S: degree of deactylation 81%, MW
50-1000 kDa, or Chitopharm L: degree of deactylation 80%, MW
500-5000 kDa.
[0056] In one embodiment, the chitosan exhibits a molecular weight
distribution from about 50 kD to about 5000 kD. A commercially
available example for such a chitosan is Chitopharm.RTM. L from the
company Cognis.
[0057] The term "molecular weight distribution" as used in the
present invention refers to a range or distribution of the
molecular weights of a population of molecules, which are not
homogeneous with respect to molecular size and weight, and which
thus can best be described by a range of molecular weights
characterized by a lower and an upper limit, where such range
covers about at least 60%, particularly at least 70%, more
particularly at least 80%, and most particularly at least 90% of
all molecular weights present in a given sample.
[0058] The chitosan beads are present in the filler at a
concentration from about 10% (v/v) to about 95% (v/v) of total
volume of the filler, as determined by determining the bead volume
after sedimentation of the bead suspension in a graduated cylinder.
In one embodiment, the chitosan beads are present in the filler at
a concentration from about 25% to about 95% of total volume of the
filler. In one further embodiment, the chitosan beads are present
in the filler at a concentration from about 50% to about 95% of
total volume of the filler. In another embodiment, the chitosan
beads are present in the filler at a concentration from about 75%
to about 95% of total volume of the filler. In one further
embodiment, the chitosan beads are present in the filler at a
concentration from about 85% to about 90% of total volume of the
filler. The amount of chitosan beads present in the filler varies
according to the size of the beads, size of the injection needle
and the location of treatment.
[0059] The term "about" as used in the present invention refers to
a 10% deviation from the value it is attached to.
[0060] Surprisingly, the chitosan beads prepared according to the
process of the instant invention are flexible and elastic in terms
of their physical properties, thus enabling an improved
syringeability.
[0061] According to the present invention, the chitosan beads
exhibit a particle size, measured as mass median diameter by
microscopical analysis or with laser diffraction, of less than or
equal to about 1500 .mu.m. In a particular embodiment, the chitosan
beads have a mass median diameter of between about 20 and 1000
.mu.m, more particularly between about 20 and 500 .mu.m, and most
particularly between about 30 and 300 .mu.m. The particle size can
be reduced by employing known techniques, such as Air jet/Air
stripping method, Jet cutter method, Vortex bowl atomizer,
Vibrating nozzle device, Electrostatic device, Emulsification
("water in oil") approach, low mid and high pressure homogenization
approaches. The size of the particle is adjusted according to the
location of treatment. After the filler is injected the size of the
chitosan beads provides fixation at the injection location and
prevents undesirable migration to other parts of patient's
body.
[0062] According to the instant invention the filler may comprise a
medium in which the chitosan beads are suspended. Said medium may
be sterile water, phosphate-buffer saline (PBS), ringer solution,
isotonic saline solution (0.9%), trometamol, citrate, carbonate,
acetate, borate, amino acids, diethylamine, glucono delta lactone,
glycine, lactate, maleic, methanesulfonic, monoethanolamine,
tartrate buffer of choice or any combination thereof. Said buffer
may be citrate buffer.
[0063] In a particular embodiment, the filler according to the
present invention does not comprise a chitosan gel.
[0064] The chitosan beads and/or filler as claimed in the instant
invention may further comprise one or more active pharmaceutical
ingredient selected from the group of anesthetics, analgesics,
anti-microbials, anti-inflammatory drugs, growth factors, hormones,
cosmeceuticals, vitamins, nutrients, stimulants, steroids,
vasoconstrictors, anti-thrombotic agents, anti-coagulation agents,
tranquilizers, muscle relaxants, antifungals, lipolytic agents and
biorejunevation agents.
[0065] The term "active pharmaceutical ingredient" refers to all
structures, which are pharmacologically active, thus resulting in a
pharmacological effect in mammal and all known chemical forms
thereof. Examples are, but not limited to, conjugates, isomers,
esters, derivatives, metabolites, residues, salts or prodrugs
thereof.
[0066] Anesthetics may be, but are not limited to, local
anesthetics based on esters (Procaine, Benzocaine, Chloroprocaine,
Cocaine, Cyclomethycaine, Dimethodcaine, Larocaine, Propoxycaine,
Proparacaine, Tretracaine) or local anesthetics based on amides
(Lidocaine, Articaine, Bupivacaine, Carticaine, Cinchocaine,
Etidocaine, Levobupivacaine, Mepivacaine, Piperocaine, Prilocaine,
Ropivacaine, Trimecaine). A suitable concentration for the
anesthetic is from about 0.01% to 6% based on the total weight of
the composition and the agent selected.
[0067] Analgesics may be, but are not limited to, paracetamol,
ibuprofen, diclofenac, naproxen, aspirin, celecoxib, etoricoxib,
lumiracoxib, parecoxib, rofecoxib, valdecoxib, nimesulid.
[0068] Antimicrobials may be, but are not limited to, antibiotics
(amikacin, gentamycin, neomycin, tobramycin, kanamycin, meropenem,
imipenem, cefaclor), antivirals (abacavir, aciclovir, amantadine,
boceprevir, cidofovir, darunavir, edoxudine, famciclovir,
ganciclovir, imunovir, inosine, interferon, lamivudine, nexavir,
oseltamivir, penciclovir, ribavirin, rimantadine, viramidine,
zidovudine) and antifungals (Miconazole, ketoconazole,
itraconazole, clotrimazole, econazole, fluconazole, voriconazole,
abafungin, naftifine, caspofungin, micafungin, benzoic acid,
griseofulvin).
[0069] Anti-inflammatory drugs may be, but are not limited to, zinc
salts, including zinc salts of polysaccharide acids, such as
hyaluronic acid.
[0070] In one embodiment of the instant invention, the one or more
active pharmaceutical ingredients are entrapped in the chitosan
beads.
[0071] In one embodiment of the instant invention, living cells,
e.g. autologous stem cells, are entrapped in the chitosan
beads.
[0072] In one embodiment of the instant invention, a polysaccharide
is entrapped in the chitosan beads.
[0073] In one embodiment of the instant invention, proteins or
peptides, e.g. adhesion proteins, granulocyte-colony stimulating
factors, erythropoietin, bone morphogenic protein, tissue
plasminogen activator, are entrapped in the chitosan beads.
[0074] In one further embodiment of the present invention, the
filler further comprises one or more pharmaceutical excipients
selected from antioxidants, viscosity enhancers/modifiers,
hydrating agents, bulking substances, tonicity agents,
preservatives and surface active agents, or a mixture thereof.
[0075] Antioxidants may be, but are not limited to, vitamin E,
vitamin C, glutathione coenzyme Q, resveratrol, bisulfite sodium,
butylated hydroxyl anisole/toluene, cysteinate, dithionite sodium,
gentisic acid, glutamate, formaldehyde sulfoxylate sodium,
metabisulfite sodium, monothiogylcerol, propyl gallate, sulfite
sodium, thiogycolate sodium, flavonoids, catalase, lycopene,
carotenes, lutein, superoxide dismutase and peroxidases or mixtures
thereof.
[0076] Viscosity enhancers may be, but are not limited to,
glycerol, xanthene gum, polyethylene glycol (PEG), alginate,
carbomers, cellulose derivatives, dextrans, and carrageenan,
starches, gum, acacia, tragacanth, gelatin, polyvinylpyrrolidone,
albumin, dextran or mixtures thereof.
[0077] Surface active agents may be, but are not limited to,
polysorbate 20, polysorbate 80, polysorbate 40, polysorbate 60,
polysorbate 65, Pluronic F68, Cetrimoniumbromid,
Cetylpyridiniumchlorid, Brij 72, Brij 30, Brij 35, deoxycholate,
lecithine, tocopheryl polyethylene glycol succinate or mixtures
thereof.
[0078] Bulking substances or tonicity modifiers may be substances
such as glycerol, lactose, mannitol, dextrose, sodium or potassium
chloride, sodium sulphate and sorbitol, in general at a
concentration up to 5% depending upon the chosen substance and the
composition of the formulation.
[0079] In certain embodiments of the present invention, the
chitosan beads are stable in form, such form stability being
determined microscopically by recording the changes in spherical
shape.
[0080] The stability is further determined by measuring
periodically the mass median diameter of the chitosan beads. In one
embodiment of the invention, the mass median diameter of the
chitosan beads remains constant, i.e. within +/-20% of the starting
value for the mass median diameter, for a period of at least 6
months, particularly at least 12 months, more particularly at least
24 months, and most particularly at least 36 months, at 25.degree.
C..+-.2.degree. C. and 60%.+-.5% relative humidity determined by
laser diffraction technique.
[0081] In one further embodiment, the filler has a shelf-life at
25.degree. C..+-.2.degree. C. and 60%.+-.5% relative humidity of at
least 6 months, particularly at least 12 months, more particularly
at least 24 months, and most particularly at least 36 months.
[0082] In certain embodiments, the chitosan beads, including those
of the filler, according to the instant invention have an
elasticity greater 5%, a tensile strength lower 5 N, and/or a
deformability greater 90%. Deformability and elasticity are
determined according to the method described by Edwards-Levy et.
al. (Biomaterials 20 (1999) 2069-2084) using a texture
analyzer.
[0083] In certain embodiments, the filler of the instant invention
is for use for aesthetic purposes.
[0084] In the context of the present invention, the term "use for
aesthetic purposes" refers to non-medical uses.
[0085] In one embodiment of the present invention, the filler is a
dermal filler
[0086] In another embodiment, the dermal filler is for the
treatment of wrinkles and/or folds.
[0087] In the context of the present invention, the term "treatment
of wrinkles and/or folds" refers to non-medical treatments.
[0088] Wrinkles that may be treated by employing the filler
according to the instant invention include, but are not limited to,
frown lines, medium depth wrinkles, such as nasolabial folds, lip
augmentation, forehead wrinkles, glabellar lines, obvious mild to
moderate nasal furrows and cheek wrinkles, crow's feet, perioral
wrinkles and acne scars.
[0089] In another embodiment of the present invention, the filler
is for the treatment of, or for use in the treatment of, a medical
condition, including lipoatrophy, gastroesophageal reflux disease
(GERD), urine incontinence, vesico ureteral reflux (VUR), and the
treatment of a psychological condition caused by the appearance of
an aesthetic deficiency, including, but not limited to, frown
lines, medium depth wrinkles, such as nasolabial folds, lip
augmentation, forehead wrinkles, glabellar lines, obvious mild to
moderate nasal furrows and cheek wrinkles, crow's feet, perioral
wrinkles and acne scars.
[0090] In another embodiment of the present invention, the filler
is for use in plastic, cosmetic, dental or general surgery, in
ophthalmology, in orthopedics, for preventing tissue adhesions, or
in urology.
[0091] The present invention further pertains to methods of using
the chitosan beads and/or the fillers of the present invention for
aesthetic purposes, including the use as dermal filler, such as in
the treatment of wrinkles and/or folds.
[0092] The present invention further pertains to methods of using
the chitosan beads and/or the fillers of the present invention for
the therapeutic treatment of a patient in need thereof, such as in
the treatment of lipoatrophy, gastroesophageal reflux disease
(GERD), urine incontinence, vesico ureteral reflux (VUR), and the
treatment of a psychological condition caused by the appearance of
an aesthetic deficiency, including, but not limited to, frown
lines, medium depth wrinkles, such as nasolabial folds, lip
augmentation, forehead wrinkles, glabellar lines, obvious mild to
moderate nasal furrows and cheek wrinkles, crow's feet and perioral
wrinkles.
[0093] The present invention further pertains to a method of
treating a medical condition, including lipoatrophy,
gastroesophageal reflux disease (GERD), urine incontinence, vesico
ureteral reflux (VUR), or a psychological condition caused by the
appearance of an aesthetic deficiency, including, but not limited
to, frown lines, medium depth wrinkles, such as nasolabial folds,
lip augmentation, forehead wrinkles, glabellar lines, obvious mild
to moderate nasal furrows and cheek wrinkles, crow's feet, perioral
wrinkles and acne scars, wherein said method comprises a step of
administering a filler as claimed in the present invention to a
patient in need thereof
[0094] The present invention further pertains to a method of using
a filler according to the present invention in plastic, cosmetic,
dental or general surgery, in ophthalmology, in orthopedics, as
products for preventing tissue adhesions, or in urology, wherein
said method comprises a step of administering a filler as claimed
in the present invention to a patient in need thereof.
[0095] The present invention further pertains to a process for
preparing chitosan beads and/or a filler according to the
invention, which comprises a step of dropping a chitosan solution
into an aqueous solution containing citrate anions.
[0096] In one embodiment, the pH of the aqueous solution containing
citrate anions is adjusted to a value from about 5 to about 10.
[0097] In one further embodiment, the concentration of the citrate
ions is below about 2.0 M.
[0098] In another embodiment, the concentration of citrate ions is
between about 0.01 M and about 1.0 M.
[0099] In another embodiment of the present invention, the
concentration of chitosan in the process for preparing chitosan
beads and/or the filler is at maximum about 5.0 wt-% (w/w %)
relative to the total weight of the composition, particularly
between about 0.5 wt-% and 4 wt-%, more particularly between about
0.5 wt-% and 3 wt-%, and most particularly between about 0.5 wt-%
and 2 wt-%.
[0100] In one further embodiment of the instant invention, the
viscosity of the chitosan solution in the process for preparing
chitosan beads and/or the filler is in the range from about 50
mPa*s to about 2000 mPa*s measured by a falling ball viscometer at
20.degree. C., particularly between about 100 mPa*s and about 1700
mPa*s, more particularly between about 500 mPa*s and about 1500
mPa*s, and most particularly between about 750 mPa*s and about 1250
mPa*s.
[0101] In one further embodiment of the process of the present
invention, the aqueous solution containing citrate ions may further
comprise one or more active pharmaceutical ingredient selected from
the group of anesthetics, analgesics, anti-microbials,
anti-inflammatory drugs, growth factors, hormones, cosmeceuticals,
vitamins, nutrients, stimulants, steroids, vasoconstrictors,
anti-thrombotic agents, anti-coagulation agents, tranquilizers,
muscle relaxants, antifungals, lipolytic agents and biorejunevation
agents.
[0102] The present invention further pertains to a kit comprising
(a) the filler as disclosed herein, and (b) an injection device. In
one embodiment, the injection device comprises a 25- to 32-gauge
needle. The size of the needle will be determined by the filler
composition, the depth of the injection site and the injection
volume. In certain embodiments, the injection device is disposable.
In one embodiment, the injection device is made of sterile
glass.
[0103] The present invention further pertains to an injection
device comprising chitosan beads and/or a filler as disclosed
herein. In one embodiment, the injection device may comprise a 25-
to 32-gauge needle. The size of the needle will be determined by
the filler composition, the depth of the injection site and the
injection volume. In certain embodiments, the injection device is
disposable. In one embodiment, the injection device may be made of
sterile glass.
[0104] In certain embodiments, the injection device and the
chitosan beads and/or filler provided herein are both sterile and
non-pyrogenic e.g. containing less than 10 EU (Endotoxin Unit, a
standard measure) per dose or application. The methods of achieving
the sterility of the chitosan beads and/or filler are those known
to the person skilled in the art.
[0105] Isotonicity of the chitosan beads and/or filler may be
accomplished by employing sodium chloride, or other
pharmaceutically acceptable agents such as dextrose.
[0106] A pharmaceutically acceptable preservative may be employed
to improve the shelf-life of the filler. The preservative may be,
but is not limited to, benzalkonium chloride, thiomersal, parabens,
chlorobutanol, benzethonium chloride, m-cresol, phenol,
2-phenoxyethanol, phenyl mercuric nitrate or benzyl alcohol. The
suitable concentration of the preservative agent is from about
0.001% to 5% based on the total weight of the composition and the
agent selected.
[0107] In another aspect, the invention relates to a method,
wherein the chitosan beads cross-linked with citrate ions are
redissolved after implantation, to the extent necessary, by the
injection of a solution containing divalent or trivalent
cations.
[0108] In another embodiment, the injection volume of the dispersed
beads is between 0.1 and 100 ml, particular between 0.1 and 50 ml,
more particular between 0.1 and 30, 0.1 and 20, or 0.1 and 10 ml,
and most particular between 0.1 and 5, 0.1 and 2, or 0.1 and 1 ml.
Alternatively, the volume can be higher than 100 ml if larger areas
are augmented.
[0109] The invention is now described with reference to the
following examples. These examples are provided for the purpose of
illustration only and the invention should not be construed as
being limited to these examples, but rather should be construed to
encompass any and all variations which become evident as a result
of the teaching provided herein. The following materials and
methods are provided with respect to the subsequent examples but do
not limit a multiplicity of materials and methodologies encompassed
by the present invention.
EXAMPLES
Example 1
Manufacturing of Chitosan Beads and/or a Filler Comprising such
Beads
[0110] Chitosan (Chitopharm L) is dispersed in deionized water. The
appropriate amount of acid is added to the dispersion under
constant stirring (0.05 mol per 10 g of chitosan). The dispersion
is stirred until a clear solution is obtained. Having obtained a
clear solution the pH of the obtained chitosan solution is
stabilized from pH 1 to 5. With the help of a 30 Gauge syringe the
chitosan solution is dropped into the following solutions having a
pH from 5 to 11 and the obtained beads are let for 1 h in
solution.
TABLE-US-00001 Crosslinking media 1M Citrate 1M citrate 1M citrate
1M citrate pH of the media 8 9 10 11 Observation Formation of
Formation of Formation of Formation of after dropping circular
beads circular beads circular beads circular beads Observation
Elastic, flexible Elastic, flexible Less flexible, Less flexible,
after 1 h crosslinking beads beads more rigid more rigid beads
beads
[0111] The obtained chitosan beads were investigated by employing a
microscope (Mikroskop Nikon Eclipse E600W). FIG. 1 shows chitosan
beads cross-linked with citrate ions and stored in citrate buffer.
The beads are perfect in shape, i.e. being circular. Bead size is
1.0 to 1.5 mm.
Example 2
Loading of Chitosan Beads with Active Pharmaceutical
Ingredients
[0112] Chitosan is dispersed in deionized water. The appropriate
amount of acid is added to the dispersion under a constant
stirring. The dispersion is stirred until a clear solution is
obtained. As a second solution, lidocaine hydrochloride USP (2%
weight per weight) is dissolved in the chitosan solution. With the
help of a 30 Gauge syringe the chitosan lidocaine solution is
dropped into the following solutions having a pH from 1 to 5 and
the obtained beads with lidocaine encapsulated are let for 1 h in
solution.
Example 3
Administration of the Filler Comprising Chitosan Beads
[0113] The filler prepared according to example 1 is injected to a
50-year old female patient into nasolabial folds.
Example 4
Determination of the Elasticity and Flexibility of Chitosan
Beads
[0114] Chitosan citrate beads were prepared according to the method
described in example 1 at pH 8.5. Ten randomly chosen beads were
selected for each experiment and the mean value and standard
deviation were calculated.
[0115] Elasticity and flexibility were determined using a Texture
Analyser TA.XT plus, Stable Micro Systems Ltd according to the
method described by Edwards-Levy et. al. (Biomaterials 20 (1999)
2069-2084) with some minor modifications. For the rupture study and
deformability study a single chitosan bead in a Petri dish was
placed under a piston, the piston went down at the rate of 1.0
mm/s, until a resistance force of 2.5 g was detected meaning the
contact of the piston with the top of the bead. Then, the piston
went down at a constant rate of 0.5 mm/s until it hit the bottom of
the Petri dish, while the force opposed to the bead as a function
of the displacement was determined. The rupture force as the
initial force recognized when the piston reached the bead was
calculated and the deformability expressed as the percentage of the
total height of the sample that the piston reached before breakage.
Tensile strength of the chitosan beads was 2.0.+-.0.3 N,
Deformability was 96.+-.2%.
[0116] Elasticity of the chitosan citrate beads was determined and
calculated as the ratio of the force opposed by the bead after 10 s
to the instantaneous resistance strength of the bead. The bead was
placed under the piston, which went down at a rate of 2.0 mm/s
until it reached 30% of the total height of the bead. Then, the
piston stayed motionless at this position for 10 s and finally
returned to its initial position. The elasticity of the beads was
7.2.+-.1.7%.
* * * * *