U.S. patent application number 16/385595 was filed with the patent office on 2020-03-12 for preventative therapy for post-traumatic osteoarthritis.
The applicant listed for this patent is University of Iowa Research Foundation. Invention is credited to Marc Brouillette, Mitchell Coleman, Tae-Hong Lim, James A. Martin, Todd O. McKinley.
Application Number | 20200078491 16/385595 |
Document ID | / |
Family ID | 56801838 |
Filed Date | 2020-03-12 |
United States Patent
Application |
20200078491 |
Kind Code |
A1 |
McKinley; Todd O. ; et
al. |
March 12, 2020 |
PREVENTATIVE THERAPY FOR POST-TRAUMATIC OSTEOARTHRITIS
Abstract
Compositions comprising a reverse-temperature sensitive hydrogel
comprising a biopolymer such as a polysaccharide and a synthetic
polymer, and a compound in an amount that reversibly inhibits
respiratory enzyme complex I, and methods of using the composition,
are provided.
Inventors: |
McKinley; Todd O.;
(Indianapolis, IN) ; Martin; James A.; (Iowa City,
IA) ; Coleman; Mitchell; (Iowa City, IA) ;
Lim; Tae-Hong; (Coralville, IA) ; Brouillette;
Marc; (Coralville, IA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of Iowa Research Foundation |
Iowa City |
IA |
US |
|
|
Family ID: |
56801838 |
Appl. No.: |
16/385595 |
Filed: |
April 16, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15895518 |
Feb 13, 2018 |
10314941 |
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16385595 |
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PCT/US2016/047360 |
Aug 17, 2016 |
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15895518 |
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62207059 |
Aug 19, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/515 20130101;
A61L 2300/434 20130101; A61L 2430/24 20130101; A61K 47/34 20130101;
A61K 9/06 20130101; A61L 27/52 20130101; A61K 9/0024 20130101; A61L
27/26 20130101; A61K 31/155 20130101; A61K 47/36 20130101; A61L
2400/06 20130101; A61L 27/54 20130101; A61P 19/02 20180101; C08L
5/08 20130101; C08L 71/02 20130101; A61L 27/50 20130101; A61K 47/38
20130101; A61L 27/26 20130101; A61L 27/26 20130101 |
International
Class: |
A61L 27/26 20060101
A61L027/26; A61K 47/38 20060101 A61K047/38; A61K 31/155 20060101
A61K031/155; A61P 19/02 20060101 A61P019/02; A61K 31/515 20060101
A61K031/515; A61K 47/36 20060101 A61K047/36; A61K 47/34 20060101
A61K047/34; A61K 9/06 20060101 A61K009/06; A61K 9/00 20060101
A61K009/00; A61L 27/54 20060101 A61L027/54; A61L 27/52 20060101
A61L027/52; A61L 27/50 20060101 A61L027/50 |
Goverment Interests
STATEMENT OF GOVERNMENT RIGHTS
[0002] This invention was made with government support under grant
W81XWH-11-1-0583 awarded by the Department of Defense. The
government has certain rights in the invention.
Claims
1. A composition comprising a reverse-temperature sensitive
hydrogel comprising a polysaccharide and a synthetic polymer, and a
compound that prevents or inhibits chondrocyte death or enhances
chondrocyte function.
2. The composition of claim 1 wherein the polysaccharide comprises
hyaluronic acid (HA), hydroxypropylcellulose, karaya gum (KG), guar
gum (GUG), or gellan gum (GEG).
3. The composition of claim 2 wherein the hydrogel includes about
0.2 wt/vol to about 4% wt/vol HA.
4. The composition of claim 2 wherein the HA has a MW of 1.5 M
Dalton or greater.
5. The composition of claim 2 wherein the HA has a MW of about
1,600,000 to 3,200,000.
6. The composition of claim 1 wherein the synthetic polymer
comprises a polaxamer.
7. The composition of claim 1 wherein the compound comprises
amobarbital or a derivative thereof.
8. The composition of claim 1 wherein the compound comprises
adenosine diphosphate ribose, or metformin.
9. A method to prevent or inhibit post-traumatic osteoarthritis
after injury in a mammal, comprising administering to an injured
joint of the mammal an effective amount of the composition of claim
1.
10. The method of claim 9 wherein the mammal is a human.
11. A method of treating articular injury in a mammal, comprising:
injecting into an injured joint of a mammal an effective amount of
the composition of claim 1.
Description
CLAIM FOR PRIORITY
[0001] This application is a continuation of and claims the benefit
of priority to U.S. patent application Ser. No. 15/895,518, filed
Feb. 13, 2018, which application is a continuation of and claims
the benefit of priority to International Application No.
PCT/US2016/047360, filed Aug. 17, 2016, which claims the benefit of
priority to U.S. Provisional Patent Application Ser. No.
62/207,059, filed Aug. 19, 2015, the disclosures of which are
incorporated by reference herein.
BACKGROUND
[0003] The pain, immobility, and general disability associated with
osteoarthritis are familiar to most people who reach old age.
Post-traumatic osteoarthritis (PTOA) is a profoundly accelerated
form of arthritis associated with traumatic injuries to joint
articular surfaces, leading to disease progression well before
patients are considered good candidates for joint replacement
approaches common to orthopaedic medicine. Because patients are
often injured relatively young and there are presently no viable
alternatives to joint replacement, patients with PTOA often suffer
disability and morbidity comparable to chronic heart disease
patients.
[0004] Natural methods for treating PTOA include decreasing load
and stress on the injured joint or increasing comfort and
functionality. For example, weight loss, low impact exercise, and
strengthening muscles surrounding the joint may improve PTOA.
However, these approaches do not cure or prevent PTOA and may not
be fully effective.
[0005] Non-steroidal, anti-inflammatory medicines (NSAIDS) are used
to decrease pain and inflammation associated with PTOA, although
NSAIDs can cause stomach irritation and kidney, liver or heart
problems. Moreover, NSAIDs likely do not prevent PTOA.
Antioxidants, another class of compounds used to treat PTOA,
stabilize or deactivate reactive oxygen species (ROS) before they
attack cells. Nevertheless, there is skepticism about the benefit
of antioxidants and there are potentially harmful side effects if
anti-oxidants are taken in excess.
[0006] Other methods used to treat PTOA include the administration
of cortisone and hylamers which act like artificial joint fluid
after injection. However, cortisone can cause elevation of heart
rate and blood sugar and should not be given too often. In
addition, cortisone is not preventative. While corticosteroid
injections are anti-inflammatory, the potential benefit or adverse
effects of that injection for traumatic injury have not been
resolved. Another approach is the use of platelet-rich plasma
injections.
[0007] Injection of a patient's own platelets leads to release of
growth factors and attraction of regenerative cells to the site of
injury. This type of injection is not preventative and does not
work for all PTOA patients. Moreover, details on dosage, frequency
of injection, and other important parameters have yet to be worked
out for platelet rich plasma administration. A further type of
injection is an amniotic membrane stem cell injection. While this
injection is anti-inflammatory, thus providing pain relief, and
results in replacement of damaged cells due to release of growth
factors, it is not preventative and does not target ROS.
[0008] If non-surgical methods are ineffective, surgical methods
may be employed to restore the joint after PTOA. The surgery may
include cleaning out, reconstructing or replacing the worn out
joint surfaces. As with other surgeries, there can be surgical
complications, e.g., infection and damage to surrounding
structures, blood clots, heart attack, and stroke, and the eventual
wearing out or loosening of implants.
SUMMARY
[0009] The present disclosure provides an injectable composite
hydrogel comprising a polysaccharide, e.g., a natural
polysaccharide such as hyaluronic acid, hydroxypropylcellulose,
karya gum (KG), guar gum (GUG), or gellan gum (GEG), a
semi-synthetic polysaccharide or a synthetic polysaccharide, and a
synthetic polymer, e.g., F127, whose reverse-thermal properties
cause the composite to become firm once injected (preventing
leakage from the site of injection such as a joint), and a compound
useful to prevent, inhibit or treat PTOA. In one embodiment, the
compound reversibly inhibits the respiratory enzyme complex I, a
key mediator of chondrocyte injury after impact. In one embodiment,
the hydrogel comprises an effective amount of amobarbital, e.g.,
from about 0.25 mM to about 50 mM or about 1.25 mM to about 10 mM,
metformin (N,N-dimethylbiguanide) a biguanide derivative,
N,N-diethylbiguanide, N,N-dipropylbiguanide, phenformin (Sogame et
al., Biopharm. Drug Dispos., 30:476 (2009)), or HL010183 (Koh et
al., Bioorg. Med. Chem., 21:2305 (2013)), or adenosine diphosphate
ribose or a derivative thereof. In one embodiment, the volume
administered is about 0.1 mL to about 15 mL, e.g., about 1 mL to
about 10 mL or about 2 mL to about 5 mL. The combination of
materials in the hydrogel offers a practical advantage, for
instance, in enabling health care providers to protect articular
tissue acutely after injury. Also, the use of compounds that
reversibly inhibit the respiratory enzyme complex I to alter
articular cartilage provides for chondroprotection after injury and
eventual reestablishment of normal activity of the respiratory
enzyme complex I.
[0010] The disclosure provides an injectable composition comprising
a composite reverse-temperature sensitive hydrogel comprising a
biopolymer, such as a polysaccharide, and a synthetic polymer, and
a compound in an amount that optionally reversibly inhibits
respiratory enzyme complex I. In one embodiment, the hydrogel
includes about 0.2 wt/vol to about 4% wt/vol HA In one embodiment,
the polysaccharide comprises hyaluronic acid. In one embodiment,
the synthetic polymer comprises a poloxamer, e.g., F127. In one
embodiment, the hydrogel includes about 15% wt/vol to about 20%
wt/vol F127. In one embodiment, the compound comprises amobarbital.
In one embodiment, the amount of the compound in the hydrogel
inhibits mitochondrial dysfunction or chondrocyte energy
dysfunction. In one embodiment, the compound scavenges
mitochondrial oxidants or prevents their formation, or stimulates
glycolytic ATP production In one embodiment, the hydrogel comprises
N-isopropyl acrylamide polymer, ethylhydroxyethylcellulose,
poly(etheylene oxide-b-propylene oxide-b-ethylene oxide),
poloxamers, PLURONICS.RTM. polymers, poly(ethylene
glycol)/poly(D,L-lactic acid-co-glycolic acid) block co-polymers,
polysaccharides, alginate, polyphosphazines, polyacrylates,
TETRONICS.TM. polymers, or polyethylene oxide-polypropylene glycol
block copolymers. In one embodiment, the polysaccharride comprises
hyaluronic acid of about or greater than 1.5 M Dalton. In one
embodiment, the MW is about 1,600,000 to 3,200,000, or about
1,900,000 to 3,900,000.
[0011] In one embodiment, the polysaccharide comprises
hydroxypropylcellulose, karya gum (KG), guar gum (GUG), or gellan
gum (GEG). In one embodiment, the polysaccharide is present in the
hydrogel at about 0.2% (wt/vol) to about 1.0% (wt/vol).
[0012] In one embodiment, the composition is a reverse
temperature-sensitive hydrogel (one that is non-viscous at "low"
temperature, e.g., at or below room temperature, e.g., about
70.degree. F. or less. The low initial viscosity allows the
hydrogel to coat all the cartilage surfaces through the joint
before it sets (i.e., the viscosity increases at temperatures above
room temperature, e.g., about 80.degree. F. or greater including
human body temperature such as about 98.degree. F.), which provides
for superior retention in the joint and substantially improves the
bioavailability of the compound dissolved in the gel. Reverse
temperature-sensitive hydrogels, which have initial viscosities of
about 100 to about 160 or about 80 to about 200, e.g., about 120 to
about 140, Pascal Seconds, may be administered using a 22 to 24
gauge needle, e.g., a 22 gauge needle. In contrast, non-reverse
temperature-sensitive hydrogels require large bore needles and do
not evenly distribute in the joint due to their high initial
viscosity.
[0013] Also provided is a method to prevent or inhibit chondrocyte
death and improve chondrocyte function after injury in a mammal.
The method includes administering an effective amount of the
composition to a mammal having the injury. Further provided is a
method to prevent or inhibit post-traumatic osteoarthritis in a
mammal. The method includes administering an effective amount of
the composition a mammal at risk of posttraumatic osteoarthritis.
In one embodiment, the composition comprises hyaluronic acid. In
one embodiment, the composition comprises F127. In one embodiment,
the composition comprises amobarbital. In one embodiment, the
amount administered inhibits mitochondrial dysfunction or
chondrocyte energy dysfunction. In one embodiment, the compound
administered scavenges mitochondrial oxidants or prevent their
formation, in addition to stimulating glycolytic ATP production. In
one embodiment, the administration is within 1, 2, 3, 4 or 5 days
of the injury. In one embodiment, the mammal has an injured joint.
In one embodiment, the administration is with 1, 2, 3, 4, 5, 6, 7,
8, 9, 10 11, or 12 hours of the injury.
DETAILED DESCRIPTION OF THE FIGURES
[0014] FIG. 1 is a schematic of the electron transport chain.
Electrons from donor molecules are transferred through protein
complexes. As electrons are transferred, hydrogen ions are pumped
across the inner membrane of the mitochondria, and as the hydrogen
atoms fall back over the inner membrane, they generate ATP.
[0015] FIG. 2 is a schematic of reactive oxygen species
production.
[0016] FIG. 3 is a schematic of steps in the progression to
post-traumatic osteoarthritis.
[0017] FIG. 4 shows complex I activity in the presence or absence
of amobarbital.
DETAILED DESCRIPTION
Definitions
[0018] "Hydrogel" as used herein means a water insoluble, naturally
or chemically-induced cross-linked, three-dimensional network of
polymer chains plus water that fills the voids between polymer.
Cartilage, Electron Transport and PTOA
[0019] Articular cartilage is the smooth, white tissue that covers
the ends of bones where they come together to form joints. It
allows the bones to glide over each other with very little
friction, and acts as a cushion. Injured, inflamed, or damaged
cartilage does not heal itself well due to lack of a blood supply,
resulting in symptoms such as pain and limited movement leading to
joint damage and deformity. Chondrocytes are cells found in
cartilage connective tissue they produce and maintain the cartilage
matrix. Under normal circumstances, cartilage wears down over time
and chondrocytes replace and repair it as needed.
[0020] Chondrocyes, like all cells, contain mitochondria.
Mitochondria generate ATP through the Electron Transport Chain
(ETC) (FIG. 1). Sometimes, harmful substances called reactive
oxygen species (ROS) (FIG. 2) are created through the ATP
generation process and are formed by Respiratory Complex I in the
Electron Transport Chain. ROS can act as signaling molecules and
signal healthy chondrocytes to undergo apoptosis (cell suicide),
depending on the severity and length of exposure, which leads to
osteoarthritis.
[0021] Osteoarthritis is wearing out of joint surface cartilage
over time. Post-traumatic osteoarthritis (PTOA) is wearing out of a
joint that has had any kind of physical injury. PTOA is a
debilitating consequence of intraarticular fractures. Patient
outcomes after intraarticular fractures have not improved
significantly in spite of improved surgical techniques. PTOA is
relatively common: As of 2006 approximately 12% of the overall
prevalence of symptomatic OA was attributable to PTOA of the hip,
knee, or ankle. This corresponds to approximately 5.6 million
individuals in the United States being affected by PTOA. The
corresponding aggregate financial burden specifically of PTOA is
$3.06 billion annually, or approximately 0.15% of the total U.S.
health care direct cost outlay.
Compositions and Methods to Prevent, Inhibit or Treat PTOA
[0022] Inhibiting electron transport and associated oxidant
production by chondrocytes after impact injuries associated with
PTOA prevents cell death and dysfunction. Accordingly, by muting
chondrocyte mitochondrial metabolism acutely after traumatic injury
using compounds that inhibit respiratory enzyme complex I (and also
decrease ROS) that are delivered intra-articularly in a hydrogel
vehicle, the treatment is confined to the joint capsule and
prevents leaking out of any joint disruptions present. This allows
controlled local delivery of an effective pharmaceutical in a
manner that minimizes exposure to the rest of the body. For
example, amobarbital is a barbiturate derivative used to produce
relaxation, sleep, anesthesia, and anticonvulsant effects. It
inhibits respiratory complex I, leading to a decrease in ROS.
Because the effect of amobarbital in inhibiting mitochondrial
electron transport is reversible, unlike rotenone or other more
toxic alternatives, transient manipulation of chondrocyte
metabolism in this manner can prevent chondrocyte injury and death,
as well as subsequent disease, while avoiding toxic insult to the
patient due to return of oxidative metabolism.
[0023] The present compositions and method are useful to prevent,
inhibit or treat PTOA, and may substantially lower or eliminate
treatment costs and morbidities associated with other more invasive
approaches that require multiple surgical procedures and/or cell
harvests.
Hydrogels and Polymers Useful in Hydrogels
[0024] Hydrogels can be classified as those with crosslinked
networks having permanent junctions or those with physical networks
having transient junctions arising from polymer chain entanglements
or physical interactions, e.g., ionic interactions, hydrogen bonds
or hydrophobic interactions. Natural materials useful in hydrogels
include natural polymers, which are biocompatible, biodegradable,
support cellular activities, and may include proteins like fibrin,
collagen or gelatin, and/or polysaccharides like hyaluronic acid,
starch, alginate or agarose. Synthetic polymers useful in hydrogels
are prepared by chemical polymerization and include by way of
example poloxamers, acrylic acid, hydroxyethyl-methacrylate (HEMA),
vinyl acetate, and methacrylic acid (MAA).
[0025] Various methods may be used to prepare hydrogels, e.g.,
crosslinkers, copolymerization of monomers using multifunctional
co-monomer, cross linking of linear polymers by irradiation or by
chemical compounds. Monomers contain an ionizable group that can be
ionized or can undergo a substitution reaction after the
polymerization is completed. Exemplary crosslinkers are
glutaraldehyde, calcium chloride and oxidized konjac glucomannan
(DAK).
[0026] Some classes of hydrogels include (a) homopolymeric
hydrogels which are derived from a single species of monomer.
Homopolymers may have cross-linked skeletal structure depending on
the nature of the monomer and polymerization technique; (b)
copolymeric hydrogels which are comprised of two or more different
monomer species with at least one hydrophilic component, arranged
in a random, block or alternating configuration along the chain of
the polymer network; (c) multipolymer interpenetrating polymeric
hydrogel (IPN) which is made of two independent cross-linked
synthetic and/or natural polymer components, contained in a network
form. In semi-IPN hydrogel, one component is a cross-linked polymer
and other component is a non-cross-linked polymer.
[0027] Biodegradable hydrogels as a delivery vehicle have the
advantage of being environmentally friendly to the human body (due
to their biodegradability) and of providing more predictable,
controlled release of the impregnated drugs. Hydrogels are of
special interest in biological environments since they have a high
water content as is found in body tissue and are highly
biocompatible. Hydrogels and natural biological gels have
hydrodynamic properties similar to that of cells and tissues.
Hydrogels minimize mechanical and frictional irritation to the
surrounding tissue because of their soft and compliant nature.
Therefore, hydrogels provide a far more user-friendly delivery
vehicle than the relatively hydrophobic carriers like silicone, or
vinyl acetate.
[0028] Biocompatible materials that may be present in a hydrogel
include, e.g., permeable configurations or morphologies, such as
polyvinyl alcohol, polyvinylpyrrolidone and polyacrylamide,
polyethylene oxide, poly(2-hydroxyethyl methacrylate); natural
polymers such as polysaccharides, gums and starches; and include
poly[.alpha.(4-aminobutyl)]-1-glycolic acid, polyethylene oxide,
polyorthoesters, silk-elastin-like polymers, alginate, EVAc
(poly(ethylene-co-vinyl acetate), microspheres such as poly
(D,L-lactide-co-glycolide) copolymer and poly (L-lactide),
poly(N-isopropylacrylamide)-b-poly(D,L-lactide), a soy matrix such
as one cross-linked with glyoxal and reinforced with a bioactive
filler, e.g., hydroxylapatite,
poly(epsilon-caprolactone)-poly(ethylene glycol) copolymers,
poly(acryloyl hydroxyethyl) starch, polylysine-polyethylene glycol,
or agarose.
[0029] In one embodiment, the hydrogel includes poloxamers,
polyacrylamide, poly(2-hydroxyethyl methacrylate),
carboxyvinyl-polymers (e.g., Carbopol 934, Goodrich Chemical Co.),
cellulose derivatives, e.g., methylcellulose, cellulose acetate and
hydroxypropyl cellulose, polyvinyl pyrrolidone or polyvinyl
alcohols, or combinations thereof.
[0030] In some embodiments, the hydrogel includes collagen, e.g.,
hydroxylated collagen, fibrin, polylactic-polyglycolic acid, or a
polyanhydride. Other examples include, without limitation, any
biocompatible polymer, whether hydrophilic, hydrophobic, or
amphiphilic, such as ethylene vinyl acetate copolymer (EVA),
polymethyl methacrylate, polyamides, polycarbonates, polyesters,
polyethylene, polypropylenes, polystyrenes, polyvinyl chloride,
polytetrafluoroethylene, N-isopropylacrylamide copolymers,
poly(ethylene oxide)/poly(propylene oxide) block copolymers,
poly(ethylene glycol)/poly(D,L-lactide-co-glycolide) block
copolymers, polyglycolide, polylactides (PLLA or PDLA),
poly(caprolactone) (PCL), or poly(dioxanone) (PPS).
[0031] In another embodiment, the biocompatible material includes
polyethyleneterephalate, polytetrafluoroethylene, copolymer of
polyethylene oxide and polypropylene oxide, a combination of
polyglycolic acid and polyhydroxyalkanoate, gelatin, alginate,
poly-3-hydroxybutyrate, poly-4-hydroxybutyrate, and
polyhydroxyoctanoate, and polyacrylonitrilepolyvinylchlorides.
[0032] In one embodiment, the following polymers may be employed,
e.g., natural polymers such as alginate, agarose, starch, fibrin,
collagen, gelatin, chitin, glycosaminoglycans, e.g., hyaluronic
acid, dermatan sulfate and chrondrotin sulfate, and microbial
polyesters, e.g., hydroxyalkanoates such as hydroxyvalerate and
hydroxybutyrate copolymers, and synthetic polymers, e.g.,
poly(orthoesters) and polyanhydrides, and including homo and
copolymers of glycolide and lactides (e.g., poly(L-lactide,
poly(L-lactide-co-D,L-lactide), poly(L-lactide-co-glycolide,
polyglycolide and poly(D,L-lactide), pol(D,L-lactide-coglycolide),
poly(lactic acid colysine) and polycaprolactone.
[0033] In one embodiment, the hydrogel comprises a poloxamer
(polyoxyethylene, polyoxypropylene block copolymers, e.g.,
poloxamer 127, 231, 182 or 184).
Exemplar Components for Use in Hydrogels to Prevent, Inhibit or
Treat PTOA
[0034] In one embodiment, the hydrogels useful in the compositions
and methods of the invention are synthesized from a naturally
occurring biodegradable, biocompatible, and hydrophilic
polysaccharide, and a synthetic biocompatible polymer, such as
poloxamers, polylactide ("PLA"), polyglycolide ("PGA"), or
poly(lactic acid co-glycolic acid) ("PLGA").
[0035] The composition of the invention that forms a hydrogel,
e.g., a reverse temperature-sensitive hydrogel, includes a
polysaccharide, including chemically cross linked polysaccharides
and a synthetic or natural polymer, and a compound that reversibly
inhibits complex I. One exemplary polysaccharide is hyaluronic acid
(HA), a naturally occurring co-polymer composed of the sugars
glucuronic acid and N-acetylglucosamine. Specifically, HA, also
named hyaluronan, is a high molecular weight (10.sup.5-10.sup.7 Da)
naturally occurring biodegradable polymer that is an unbranched
non-sulfated glycosaminoglycan (GAG) composed of repeating
disaccharides (.beta.-1,4-D-glucuronic acid (known as uronic acid)
and .beta.-1,3-N-acetyl-D-glucosamide). HA has an average MW of 4-5
million Da. HA can include several thousand sugar molecules in the
backbone. HA is a polyanion that can self-associate and that can
also bind to water molecules (when not bound to other molecules)
giving it a stiff, viscous quality similar to gelatin. Hylans are
cross-linked hyaluronan chains in which the carboxylic and N-acetyl
groups are unaffected. The MW of hylan A is about 6 million Da.
Hylans can be water-insoluble as a gel (e.g., hylan B).
[0036] HA's characteristics include its consistency,
biocompatibility, hydrophilicity, viscoelasticity and limited
immunogenicity. The hyaluronic acid backbone is stiffened in
physiological solution via a combination of internal hydrogen
bonds, interactions with solvents, and the chemical structure of
the disaccharide. At very low concentrations, HA chains entangle
each other, leading to a mild viscosity (molecular weight
dependent). On the other hand, HA solutions at higher
concentrations have a higher than expected viscosity due to greater
HA chain entanglement that is shear-dependent. Thus, solutions
containing HA are viscous, but the viscosity is tunable by varying
HA concentration and the amount of cross-linking. In addition to
the unique viscosity of HA, the viscoelasticity of HA is another
characteristic resulting from entanglement and self-association of
HA random coils in solution. Viscoelasticity of HA can be tied to
molecular interactions which are also dependent on concentration
and molecular weight.
[0037] Exemplary HA solutions for injection are shown in Table 1,
and include include Synvisc.RTM. (high molecular weight HA due to
crosslinking), Hyalgan.RTM. (sodium hyaluronate solution), and
Orthovisc.RTM. (one of the viscosupplements with the highest HA
concentration, which has lower viscosity than Synvisc.RTM.) (the
properties of those are shown in Table 2).
TABLE-US-00001 TABLE 1 Brand name (Generic name) Molecular weight
(kDa) Durolane .RTM. (Hyaluronic acid, 2%) 1000 Fermathron .RTM.
(Sodium hyaluronate, 1%) 1000 Hyalgan .RTM. (Sodium hyaluronate,
1%) 500-730 NeoVisc .RTM. (Sodium hyaluronate, 1%) 1000 Orthovisc
.RTM. (Sodium hyaluronate, 1%) 1000-2900 Ostenil .RTM. (Sodium
hyaluronate, 1%) 1000-2000 Supartz .RTM. (Sodium hyaluronate, 1%)
620-1170 Suplasyn .RTM. (Sodium hyaluronate, 1%) 500-730 Synvisc
.RTM. (Hylan G-F 20; Crosslinked HA) 6000-7000
TABLE-US-00002 TABLE 2 Viscoelastic properties Molecular Elastic
modulus Viscous modulus weight (G') (Pa) (G'') (Pa) Brand name
(kDa) at 2.5 Hz at 2.5 Hz Hyalgan .RTM. 500-730 0.6 3
(Uncrosslinked) Orthovisc .RTM. 1000-2900 60 46 (Uncrosslinked)
Synvisc .RTM. 6000-7000 111 .+-. 13 25 .+-. 2 (Crosslinked
polymer)
[0038] Dextran is another polysaccharide and is formed primarily of
1,6-.alpha.-D-glucopyranosyl residues and has three hydroxyl groups
per glucose residue that could provide greater flexibility in the
formulation of hydrogels. Dextran has been widely used for many
biomedical purposes, such as plasma expander and controlled drug
delivery vehicle, because of its highly hydrophilic nature and
biocompatibility. It is also possible to incorporate dextranase in
order to facilitate biodegradation of dextran for the meeting of
specific clinical needs.
[0039] In one embodiment, the hydrogel comprises a poloxamer.
Poloxamers are nonionic triblock copolymers composed of a central
hydrophobic chain of polyoxypropylene (poly(propylene oxide))
flanked by two hydrophilic chains of polyoxyethylene (poly(ethylene
oxide)) (.alpha.-Hydro-.omega.-hydroxypoly (oxyethylene).sub.a poly
(ocypropylene).sub.b poly (olxyethylene).sub.a block copolymer,
with two hydrophilic chains of ethylene oxide chains (PEO) that
sandwich one hydrophobic propylene oxide chain (PPO) giving a
chemical formula
HO(C.sub.2H.sub.4O).sub.a(C.sub.3H.sub.6O).sub.b(C.sub.2H.sub.4O).sub.a).
For example, poloxamer 407 is a triblock copolymer consisting of a
central hydrophobic block of polypropylene glycol flanked by two
hydrophilic blocks of polyethylene glycol. Exemplary poloxamers
include but are not limited to polyethylene-propylene glycol
copolymer, e.g., Supronic, Pluronic or Tetronic a non-ionic
triblock copolymer.
[0040] The common representation of Poloxamer is indicated as `P`
succeeded by three digits where the first two digits are to be
multiplied by 100 and that gives the molecular mass of the
hydrophobic propylene oxide and the last digit is to be multiplied
by ten that gives the content of hydrophilic ethylene oxide in
percentage.
[0041] Poloxamers usually have an efficient thermoreversible
property with characteristics sol-gel transition temperature. Below
the transition temperature it is present as a solution and above
this temperature the solution results in interaction of the
copolymer segment which leads to gelation. Poloxamers are non-toxic
and non-irritant.
TABLE-US-00003 TABLE 3 Average Ethylene Propylene molecular Weight
% of oxide oxide mass Oxyethylene Physical units (n).sup.a units
(n).sup.a PhEur 2005; PhEur USPNF Poloxamer Pluronic form (a) (b)
USPNF 23 2005 23 124 L 44 Liquid 10-15 18-23 2090-2360 44.8-48.6
46.7 .+-. 1.9 188 F 68 Solid 75-85 25-40 7680-9510 79.9-83.7 81.8
.+-. 1.9 237 F 87 Solid 60-68 35-40 6840-8830 70.5-74.3 72.4 .+-.
1.9 338 F108 Solid 137-146 42-47 12700-17400 81.4-84.9 83.1 .+-.
1.7 407 F127 Solid 95-105 54-60 9840-14600 71.5-74.9 73.2 .+-.
1.7
[0042] Compounds that reversibly inhibit complex I include but are
not limited to amobarbital or derivatives thereof, metformin or
derivatives thereof, or adenosine diphosphate ribose analogs that
disrupt NADH binding. However, non-reversible inhibitors of complex
I, e.g., Rotenone, Piericidin A or Rolliniastatin 1 and 2, in low
doses, may also have some benefit to cartilage after injury as a
result of altering ROS.
Formulations and Dosages
[0043] The components of the composition of the invention can be
formulated as pharmaceutical compositions and administered to a
mammalian host, such as a human patient in a variety of forms
adapted to the chosen route of administration. In one embodiment,
the components of the composition are locally administered to a
site of cartilage damage or suspected cartilage damage, or is
administered prophylactically.
[0044] In one embodiment, the components of the composition may be
administered by infusion or injection. Solutions may be prepared in
water, optionally mixed with a nontoxic surfactant. Dispersions may
also be prepared in glycerol, liquid polyethylene glycols,
triacetin, and mixtures thereof and in oils. Under ordinary
conditions of storage and use, these preparations contain a
preservative to prevent the growth of microorganisms.
[0045] The pharmaceutical dosage forms suitable for injection or
infusion may include sterile aqueous solutions or dispersions or
sterile powders comprising the active ingredient which are adapted
for the extemporaneous preparation of sterile injectable or
infusible solutions or dispersions. In all cases, the ultimate
dosage form should be sterile, fluid and stable under the
conditions of manufacture and storage. The liquid carrier or
vehicle may be a solvent or liquid dispersion medium comprising,
for example, water, ethanol, a polyol (for example, glycerol,
propylene glycol, liquid polyethylene glycols, and the like),
vegetable oils, nontoxic glyceryl esters, and suitable mixtures
thereof. The prevention of the action of microorganisms can be
brought about by various antibacterial and antifungal agents, for
example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal,
and the like. In many cases, it may be preferable to include
isotonic agents, for example, sugars, buffers or sodium
chloride.
[0046] Sterile injectable solutions may be prepared by
incorporating the active agent in the required amount in the
appropriate solvent with various other ingredients, as required,
optionally followed by filter sterilization. In the case of sterile
powders for the preparation of sterile injectable solutions, the
methods of preparation include vacuum drying and the freeze drying
techniques, which yield a powder of the active ingredient plus any
additional desired ingredient present in the previously
sterile-filtered solutions.
[0047] Useful solid carriers may include finely divided solids such
as talc, clay, microcrystalline cellulose, silica, alumina and the
like. Useful liquid carriers include water, alcohols or glycols or
water-alcohol/glycol blends, in which the present compounds can be
dissolved or dispersed at effective levels, optionally with the aid
of non-toxic surfactants. Adjuvants such as antimicrobial agents
can be added to optimize the properties for a given use. Thickeners
such as synthetic polymers, fatty acids, fatty acid salts and
esters, fatty alcohols, modified celluloses or modified mineral
materials can also be employed with liquid carriers to form
spreadable pastes, gels, ointments, soaps, and the like, for
application directly to the skin of the user.
[0048] Useful dosages of the compound(s) in the composition can be
determined by comparing their in vitro activity and in vivo
activity in animal models thereof. Methods for the extrapolation of
effective dosages in mice, and other animals, to humans are known
to the art; for example, see U.S. Pat. No. 4,938,949.
[0049] Generally, the concentration of the compound(s) in a liquid
composition, may be from about 0.1-25 wt-%, e.g., from about 0.5-10
wt-%. The concentration in a semi-solid or solid composition such
as a gel or a powder may be about 0.1-5 wt-%, e.g., about 0.5-2.5
wt-%.
[0050] The amount of the compound for use alone or with other
agents may vary with the type of hydrogel, route of administration,
the nature of the condition being treated and the age and condition
of the patient, and will be ultimately at the discretion of the
attendant physician or clinician.
[0051] The components of the composition may be conveniently
administered in unit dosage form; for example, containing 5 to 1000
mg, conveniently 10 to 750 mg, or conveniently 50 to 500 mg of
active ingredient per unit dosage form.
[0052] In general, however, a suitable dose may be in the range of
from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75
mg/kg of body weight per day, such as 3 to about 50 mg per kilogram
body weight of the recipient per day, for example in the range of 6
to 90 mg/kg/day, e.g., in the range of 15 to 60 mg/kg/day.
[0053] The invention will be described by the following
non-limiting example.
Example
[0054] In one embodiment, an injectable temperature-sensitive
composite hydrogel is employed where the hydrogel is liquid during
injection, then gelates when inside the body (gelates at human body
temperatures). In one embodiment, the injectable
temperature-sensitive composite hydrogel (e.g., one having
hyaluronic acid, such as Gel One which is chemically cross-linked
and has a high molecular weight, and F127) is employed to deliver a
therapeutic, for instance, the hydrogel is loaded with amobarbital
which reversibly inhibits the respiratory enzyme Complex I, a key
mediator of chondrocyte injury after impact. The hydrogel becomes
firm once injected (preventing leakage from the joint) allowing the
therapeutic to be retained in the joint region, for example, for
about 3 days after injection into the site of articular injury. In
one embodiment, the hydrogel comprises 17% (w/v) F-127 and 0.2%
(w/v) HA, and is loaded with 2.5 mM amobarbital. The
temperature-sensitive hydrogel fixes the amobarbital, which
prevents chondrocyte death, at the site of injury.
[0055] FIG. 4 shows that amobarbital directly inhibits the
biochemical activity of complex I of the electron transport chain
in chondrocytes.
REFERENCES
[0056] Martin et al., Journal of Bone and Joint Surgery, 91A: 1890.
[0057] Goodwin et al., Journal of Orthopaedic Research, 28(8):
1057. [0058] Wolff et al., Journal of Orthopaedic Research, 31:191
(2015). [0059] Sauter et al., Journal of Orthopaedic Research,
30:593. [0060] Jubeck, Arthritis Rheum., 58(9):2809.
[0061] All publications, patents and patent applications are
incorporated herein by reference. While in the foregoing
specification, this invention has been described in relation to
certain preferred embodiments thereof, and many details have been
set forth for purposes of illustration, it will be apparent to
those skilled in the art that the invention is susceptible to
additional embodiments and that certain of the details herein may
be varied considerably without departing from the basic principles
of the invention.
* * * * *