JP2007507516A - Treatment of mammalian disorders by amino sugar administration and use of amino sugar - Google Patents

Abstract

The present invention relates to treating a joint-related disorder in a mammal by administering an amino sugar.
The treatment specifically prevents, reduces or ameliorates a disease associated with a joint disorder, wherein the disease is selected from the group consisting of synovitis, subchondral bone edema and cartilage degeneration.

Description

Government Approval This invention was made in part with the support of the US government with the support of grant numbers NIH AG 07996 and AT 00052 from the National Institutes of Health. The US government may have certain rights in this invention.

This application claims priority to US Provisional Patent Application No. 60 / 507,716 filed Oct. 1, 2003 (Title of Invention: Treatment of Mammalian Disorders by Amino Sugar Administration and Use of Amino Sugars). To do.

  The present invention relates to a method of treating severe joint-related disorders in mammals by administration of amino sugars. The treatment specifically prevents, reduces or ameliorates many of the pathological markers associated with the joint disorder, which include synovitis, subchondral bone edema and cartilage degeneration. Selected from the group consisting of (cartilage degradation).

  Various pathological markers are associated with joint-related disorders and diseases. Examples of joint-related disorders and diseases include, but are not limited to, physical injury to the joint, osteoarthritis (OA), and rheumatoid arthritis. In particular, relevant pathological markers include synovitis, subchondral bone edema, and progressive cartilage degeneration, but there are many others (Ayral et al., Rheumatology, Vol. 35, 14 -17; McAlindon, Best Pract Res Clin Rheumatol 1999, 13 (2): 329-44; Ayral et al., Annals of the Rheumatic Diseases 2002, Vol. 61, suppl. 1, # OP0014; Kerin et al., Cell Mol Life Sci 2002, 59: 27-35; Hedborn et al., Cell Mol Life Sci 2002, 59: 45-53; Silver et al., Crit Rev Biomed Eng 2001, 29: 373-91; Elsaid et al., Osteoarthritis Cartilage. 2003, 11, 673-80; Altman et al., Am J Med. 1983 Oct 31, 75 (4B): 50-5).

  Unfortunately, current treatments for joint disorders are usually limited to acetaminophen, non-steroidal anti-inflammatory drugs, intraarticular corticosteroids, and hyaluronic acid, which treat general inflammation, Pain relief only slightly (Geletka et al., Best Pract Res Clin Rheumatol. 2003 Oct, 17: 791-809; Altman et al., Am J Med. 1983 Oct 31, 75 (4B): 50-5 ).

  Recently, glucosamine (GlcN) has been used to treat osteoarthritis, a joint disorder, and its mechanism of action is generally recognized as anti-inflammatory. Most recently, it has been suggested that GlcN can stop the progression of joint space stenosis and improve the biomechanics of osteoarthritic knee joints (Reginster et al., Lancet 2001, 357: 251-6; Hughes et al., Rheumatology (Oxford) 2002, 41: 279-84). Unfortunately, the optimal in vitro activity of GlcN is seen in a concentration range that is difficult or even impossible to achieve with oral administration of sugar, and GlcN is cytotoxic to chondrocytes even at low millimolar concentrations. Therefore, treatment with high concentrations of GlcN is not preferred (Sandy et al., Arch Biochem Biophys 1999, 367: 258-64; Shikhman et al., J Immunol 2001, 166: 5155-60; Setnikar et al., Arzneimittelforschung 2001 , 51: 699-725; Adebowale et al., Biopharm Drug Dispos 2002, 23: 217-25; Aghazadeh-Habashi et al ,. J Pharm Pharm Sci 2002, 5 (2): 181-4; de Mattei et al. , Osteoarthritis Cartilage 2002, 10: 816-25).

  N-acetylglucosamine (GlcNAc) is also used for the treatment of OA and RA. The results show that GlcNAc, in contrast to GlcN, is not toxic to human articular chondrocytes and does not induce chondrocyte death at concentrations higher than 50 mM (de Mattei et al., Osteoarthritis Cartilage 2002 , 10: 816-25).

  There are many patents relating to the use of GlcN and GlcNAc for the treatment of OA and / or RA. US Pat. No. 3,683,076 (Rovati) discloses the use of GlcN salts for the treatment of OA and RA, while US Pat. No. 4,870,061 (Speck) modifies by oral administration. The use of GlcNAc to treat joint diseases is disclosed, and US Pat. Nos. 5,840,715 and 6,136,795 (both Florio) provide dietary supplements to reduce arthritis. The use of GlcNAc as an agent is disclosed.

  However, to date, there is no effective means for treating, preventing or reducing severe synovitis, subchondral bone edema, and cartilage degeneration. Accordingly, there is an increasing need for the treatment, prevention and alleviation of such serious diseases related to joint disorders.

  Amino sugars have been found to reduce, prevent and ameliorate diseases associated with joint disorders. Thus, this finding provides a specific treatment for joint disorders in which one or more of such diseases exists. This newly discovered method of treating joint disorders having one or more of the above pathological markers is a targeted approach, and by the effect of the newly found amino sugar, amino sugar therapy It is possible to treat joint disorders that have not been received.

  The present invention relates to treating a joint-related disorder in a mammal by administering an amino sugar. The treatment specifically prevents, reduces or ameliorates a disease associated with joint disorders, wherein the disease is selected from the group consisting of synovitis, subchondral bone edema and cartilage degeneration.

  Preferred embodiments of the present invention treat amino sugars (including but not limited to N-acetylglucosamine, glucosamine, galactosamine, N-acetylgalactosamine, iminocyclitol, pharmaceutically acceptable salts thereof, etc.) The present invention relates to a method for preventing, reducing or ameliorating a serious disease associated with joint disorders by administering an effective amount to a mammal. In one aspect of this preferred embodiment, joint damage is determined by a particular pathological marker, and if such marker is present, a therapeutically effective amount of an amino sugar is administered. In another aspect of this preferred embodiment, joint disorders known in the art to have such pathological markers associated with joint disorders are treated with a therapeutically effective amount of an amino sugar. Preferably, a therapeutically effective amount of amino sugar is administered intra-articularly to the mammal. Also preferably, the amino sugar is GlcNAc, more preferably the amino sugar is GlcNAc contained in a matrix as a controlled release formulation.

  In another preferred embodiment of the invention, GlcNAc is administered intra-articularly to a mammal having a joint disorder to treat cartilage degeneration, subchondral bone edema, and synovitis. Preferably, the therapeutic effect is seen at the macroscopic and microscopic levels. Preferably, the therapeutic effect is, in particular, delayed cartilage degeneration, suppression of hyperplasia of subchondral bone marrow edema, and suppression of membrane inflammation in synovitis.

  Another preferred embodiment of the present invention is that a composition comprising a therapeutically effective amount of an amino sugar (preferably GlcNAc) is administered to a mammal alone or in combination with an existing anti-inflammatory or hexosaminidase inhibitor. It is a method to do. Preferably, methods for administering the formulations of the present invention include, but are not limited to, intra-articular, local, and intramuscular methods. More preferably, a controlled release formulation of amino sugar is administered intra-articularly to a mammal in need of such treatment.

  Thus, the present invention provides a method for specifically treating joint disorders having one or more pathological markers that respond favorably to amino sugar therapy. The present invention also provides a new use of amino sugars in the targeted treatment of joint disorders having one or more pathological markers. The present invention further provides compounds useful for targeted treatment of joint disorders having one or more pathological markers and pharmaceutical formulations thereof.

Abbreviations and Terminology In the present invention and specification, the following terms and abbreviations are defined with the meanings set forth below, unless otherwise specified, but these descriptions are intended to be exemplary only. However, the meaning of the term is not limited to the meaning described or mentioned throughout the specification. Rather, these descriptions are intended to include any additional aspects and / or exemplifications of terms used in the specification and claims.
Abbreviations described herein are as follows.
ACL = anterior cruciate ligament ACLT = anterior cruciate ligament cutting GlcN = glucosamine GaG = glycosaminoglycan GlcNAc = N-acetylglucosamine HA = hyaluronic acid IL-1β = interleukin-1β
IL-6 = interleukin-6
NSAID = non-steroidal anti-inflammatory drug OA = osteoarthritis PBS = phosphate buffered saline PEG = polyethylene glycol PMSF = phenylmethylsulfonyl fluoride RA = rheumatoid arthritis

  “Active ingredient” means a therapeutically effective amount of a drug or formulation thereof. Preferably, the active ingredient of the present invention is an amino sugar, more preferably the amino sugars GlcNAc and GlcN, most preferably the amino sugar GlcNAc.

  “Therapeutically effective amount” means the amount of active ingredient necessary to induce one or more of the desired pharmacological effects of the present invention. This amount can vary greatly depending on the effectiveness of the particular active ingredient, the age and weight of the individual, the response, and the nature and severity of the individual's symptoms. Therefore, there is no upper or lower limit for the amount of active ingredient. The therapeutically effective amount used in the present invention can be readily determined by one skilled in the art.

  “Alginic acid gel” refers to a natural polysaccharide polymer comprising 1,4-linked β-D-mannuronic acid residues and α-L-guluronic acid residues in different ratios. Alginate can form a stable gel particularly in the presence of specific divalent cations (calcium, barium, strontium, etc.).

“Amino sugar” means any synthetic sugar or natural sugar in which one or more carbon atoms are substituted with an amino group (—NH ₂ ). Such substitution can occur regardless of the orientation or configuration of the asymmetric carbon present in the sugar. Unless otherwise specified, “amino sugar” means any anomer (α or β) of a cyclic amino sugar. The amino sugar may be N-substituted with an alkyl group or an acyl group (that is, one hydrogen atom of the pendant amino group may be substituted with an alkyl group or an acyl group (—COR, R is lower alkyl)). . In one preferred embodiment of the present invention, R in -COR is methyl (-CH _3).

  “Arthritis” means any of the specific diseases characterized by joint inflammation, but the cause of joint inflammation can vary under various conditions. Relatively common joint diseases include rheumatoid arthritis, juvenile arthritis, ankylosing spondylitis, psoriatic arthritis and osteoarthritis.

  “Articular cartilage” or “cartilage” means a substance that covers the end of bone and forms an articular surface. Cartilage can withstand compressive forces and forms a low friction surface on which the joint can slide. Articular cartilage consists of chondrocytes and a matrix that further includes proteins and glycosaminoglycan polysaccharides.

  “Cartilage degeneration” means degeneration of tissue containing cartilage.

  “Chitin” means β-1,4-linked (poly) GlcNAc. Chitin is found throughout nature, for example, in the exoskeleton of insects and crustaceans.

  “Chitosan” means deacylated chitin or β-1,4-linked (poly) N-glucosamine.

  “Chondrocyte” means a cell present in articular cartilage. Chondrocytes produce collagen (gelatinous protein) and proteoglycans (glucosamming licans (also called mucopolysaccharides) bound to proteins).

  “Joint disorder” or “joint disorder” refers to any disease affecting a mammal's joints and indicates one or more of the following disorders: synovitis, subchondral bone edema and cartilage degeneration. .

  “Encapsulation efficiency” means the amount of compound or active ingredient encapsulated, blended, loaded, bound, fixed or incorporated in an injectable polymer gel, liposome, microsphere, nanoparticle or the like. In general, “yield” is expressed as the encapsulation rate of the active ingredient.

  “Uptake” or “encapsulation” means any method of formulating the active ingredient, which limits the free dissolution of the active ingredient in the matrix (solution, solid phase, etc.), Suppressed or inhibited. Preferred examples of active ingredient uptake or encapsulation include, but are not limited to, formulations incorporated in a matrix selected from particles, implants or gels.

  “Matrix” means a solid, gel, or liquid composition that can incorporate amino sugars and other substances (such as anti-inflammatory agents) as needed.

  “Glycosaminoglycan” refers to a long heteropolysaccharide molecule comprising repeating disaccharide units. The disaccharide unit can be composed of a modified amino sugar (D-, N-acetylgalactosamine or D-GlcNAc) and uronic acid (D-glucuronate, L-iduronate, etc.). GAG functions particularly as a lubricant within the joint. Specific physiologically important GAGs include hyaluronic acid, dermatan sulfate, chondroitin sulfate, heprin, heparan sulfate, and keratan sulfate.

“Hyaluronic acid” means a natural mucopolysaccharide containing alternating D-glucuronic acid and DN-acetylglucosamine subunits. Hyaluronic acid is a linear polysaccharide (long chain) formed by repeating disaccharide units consisting of D-glucuronic acid β (1-3) N-acetyl-D-glucosamine linked by β (1-4) glycosidic bonds. Biopolymer). Hyaluronic acid is commercially available in various molecular weight ranges from about 50,000 daltons to about 8 × 10 ⁶ daltons. Hyaluronic acid is also available as a sodium salt, which is a dry high purity material. Sodium hyaluronate can be stored with various preservatives known in the art, including, but not limited to, alkyl-substituted benzoates and alcohols, conjugates, blends and mixtures thereof.

  “Hyaluronan” means a polymer composed of repeating molecules of N-acetylglucosamine and glucuronic acid.

  “IL-1β” means interleukin-1β, that is, an immunomodulator that mediates a wide range of immune responses and inflammatory responses (such as activation of B cells and T cells).

  “Injectable formulation” means a sterile injectable composition prepared as a solution or suspension. A solid formulation suitable for dissolution or suspension in a liquid vehicle prior to injection can also be prepared. The preparation may be emulsified or may incorporate active ingredients. Injectable preparations may also contain various preservatives known in the art, including but not limited to alkyl-substituted benzoates and alcohols, conjugates, blends and mixtures thereof.

  By “injectable polymer gel” is meant a polymer matrix carrier used to incorporate or encapsulate the active ingredients of the present invention. In the case of polymer-based injectable preparations, drug dosage and timing can be adjusted by selecting and formulating various combinations of active ingredients and polymers. Total drug dose and release rate are adjustable variables. For example, drug delivery parameters can be optimized by varying solvent amount, copolymer ratio or molecular weight, and polymer solvent polarity. Polymer-based systems can also increase the lifetime of the active ingredient. By using a polymer system comprising polylactide and lactide-glycolide copolymer in the formulation, benefits such as biocompatibility and biodegradability can be obtained. Injectable polymer gels can be prepared according to processes known in the art, eg, treated, mixed, filtered, heated or sterilized.

  “Microsphere” means a polymer matrix carrier used to incorporate or encapsulate the active ingredients of the present invention. In the case of microsphere-based preparations, drug dosage and timing can be adjusted by selecting and formulating various combinations of active ingredients and polymers. Total drug dose and release rate are adjustable variables. For example, drug delivery parameters can be optimized by changing the copolymer ratio or copolymer molecular weight. Microsphere-based systems can also increase the lifetime of active ingredients. Benefits such as biocompatibility and biodegradability can be obtained by using microspheres containing lactide-glycolide copolymer in the formulation. Microspheres can be prepared, eg, processed, processed, ground, crushed, or extruded according to processes known in the art.

  “Intra-joint” refers to a method of delivering a drug directly to a joint. Conventional drug delivery routes such as oral, intravenous and intramuscular administration rely on synovial blood flow to deliver the drug to the joint, but this is inefficient. That is, transynovial transfer of small molecules from the synovial capillaries to the joint cavity generally occurs by passive diffusion, and therefore becomes less efficient as the size of the target molecule increases. Thus, access of a molecule (eg, GlcN) to the joint space is substantially limited. Avoid these limitations by intra-articular injection or perfusion of drugs.

  “Polymer” means hyaluronic acid, polyethylene glycol, a copolymer of polyethylene glycol and poly (lactic acid / glycolic acid), a polymer of lactic acid, a copolymer of poly (ethylene glycol-y- (DL-lactic acid-co-glycolic acid), It means alginic acid gel, chitosan, or a pharmaceutically acceptable salt thereof.

  “Sustained release” means a period during which a drug is released and effective, or a period during which physiological intake of the drug is effective. An induction period where little or no drug is released may precede the sustained release period, which is biphasic, ie, an initial period during which some drug is released and more drug is released. And a second period. On the other hand, “continuous release” appears to be monophasic and is used only to describe a release profile having a smooth curvilinear release time profile. One skilled in the art will appreciate that the release profile may actually correspond to an exponential or logarithmic time-release profile.

  “Synovitis” means inflammation of the intima (synovium). Synovitis is present in a variety of joint-related disorders, including but not limited to osteoarthritis, physical or traumatic injury, rheumatoid arthritis, and other autoimmune disorders.

  Amino sugars have been found to reduce, prevent and ameliorate diseases associated with joint disorders. Thus, this finding provides a specific treatment for joint disorders in which one or more of these diseases exists. This newly discovered method of treating joint disorders with one or more of the above pathological markers is a targeted approach and does not receive amino sugar therapy due to the effect of the newly discovered amino sugar The treatment of joint disorders is possible. Note that although “disease” and “pathological marker” are used interchangeably herein, these terms refer to synovitis, subchondral bone edema and cartilage degeneration.

  The present invention relates to treating a joint-related disorder in a mammal by administering an amino sugar. The treatment specifically prevents, reduces or ameliorates a disease associated with joint disorders, wherein the disease is selected from the group consisting of synovitis, subchondral bone edema and cartilage degeneration.

  Preferred embodiments of the present invention treat amino sugars (including but not limited to N-acetylglucosamine, glucosamine, galactosamine, N-acetylgalactosamine, iminocyclitol, pharmaceutically acceptable salts thereof, etc.) The present invention relates to a method for preventing, reducing or ameliorating a serious disease associated with joint disorders by administering an effective amount to a mammal. In one aspect of this preferred embodiment, joint damage is determined by a particular pathological marker, and if such marker is present, a therapeutically effective amount of an amino sugar is administered. In another aspect of this preferred embodiment, joint disorders known in the art to have such pathological markers associated with joint disorders are treated with a therapeutically effective amount of an amino sugar. Preferably, a therapeutically effective amount of amino sugar is administered intra-articularly to the mammal. Also preferably, the amino sugar is GlcNAc, more preferably the amino sugar is GlcNAc contained in a matrix as a controlled release formulation.

  In another preferred embodiment of the invention, GlcNAc is administered intra-articularly to a mammal having a joint disorder to treat cartilage degeneration, subchondral bone edema, and synovitis. Preferably, the therapeutic effect is seen at the macroscopic and microscopic levels. Preferably, the therapeutic effect is, in particular, delayed cartilage degeneration, suppression of hyperplasia of subchondral bone marrow edema, and suppression of membrane inflammation in synovitis.

  Another preferred embodiment of the present invention is that a composition comprising a therapeutically effective amount of an amino sugar (preferably GlcNAc) is administered to a mammal alone or in combination with an existing anti-inflammatory or hexosaminidase inhibitor. It is a method to do. Preferably, methods for administering the formulations of the present invention include, but are not limited to, intra-articular, local, and intramuscular methods. More preferably, a controlled release formulation of amino sugar is administered intra-articularly to a mammal in need of such treatment.

  Thus, the present invention provides a method for specifically treating joint disorders having one or more pathological markers that respond favorably to amino sugar therapy. The present invention also provides a new use of amino sugars in the targeted treatment of joint disorders having one or more pathological markers. The present invention further provides compounds useful for targeted treatment of joint disorders having one or more pathological markers and pharmaceutical formulations thereof.

  All patents, publications and patent applications cited herein are hereby incorporated by reference in their entirety. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Typical methods and materials will be described later. However, variations of the present invention can be obtained using methods or materials similar or equivalent to those described herein. The materials, methods, and examples described below are merely illustrative and not limiting.

  The following examples are intended to illustrate specific embodiments of the present invention and are not intended to limit the scope of the invention in any way.

  The post-traumatic joint deformity ACLT (anterior cruciate ligament amputation) model is one of the most widely used models for studying degenerative changes in articular cartilage. The ACLT is described in Settonet al., Osteoarthritis Cartilage 1999, 7: 2-14. ACLT is found in abnormal knee biomechanics such as human knees suffering from increased anterior drawers during extension and 90 ° flexion and joint disorders (traumatic injury, arthritis, etc.) Causes a similar increase in adduction.

  Osteoarthritis is just one of many joint disorders that exhibit one or more of the following diseases: synovitis, subchondral bone edema and cartilage degeneration. Other joint disorders include, but are not limited to, physical or traumatic injury and rheumatoid arthritis. Hereinafter, “experimental OA” used in connection with the discussion of the ACLT model does not limit the present invention to osteoarthritis. If anything, “experimental OA” is just a general term in the art. The present invention is useful for all types of joint disorders associated with the diseases described above.

  To study the effect of amino sugars on pathological markers associated with joint damage, experimental OA was induced in rabbit knees by ACLT. In these rabbits, the most severe part of cartilage degeneration occurs in the medial femoral condyle, followed by the lateral femoral condyle (Chang et al., Osteoarthritis Cartilage 1997, Sep; 5: 357-72). In the tibial plateau, ACL cuts only cause mild to moderate lesions in the area covered by the meniscus.

Reagent GlcNAc was purchased from Sigma (St. Louis, MO). GlcNAc was dissolved in normal saline and sterilized by filtration through a 0.22 μm filter (Corning, Acton, Mass.). GlcNAc sterile solution was stored at 4 ° C. Sodium hyaluronate (Hyalgan ™) was purchased from Sanofi Synth Lab (New York, NY).

Preparation of GlcNAc sustained release preparation Polylactic acid depot (PLAD)
USP lyophilized cGMP grade GlcNAc (Greenfield, Gumee, Ill.) Powder, pharmaceutical grade low molecular weight polylactic acid (L-102, Boehringer Ingelheim (BI) Chemicals, Wallingford, Conn., USA) / NF grade solvent (benzyl alcohol (BA), benzyl benzoate (BB), ethanol (EtOH)) dissolved or suspended in a polymer solution obtained. The resulting mixture was evaluated in vitro and in vivo.

Polylactic acid-co-glycolic acid (PLGA) injectable gel Lyophilized GlcNAc powder, pharmaceutical grade low molecular weight PLGA (RG 502-H, Boehringer Ingelheim (BI) Chemicals, Wallingford, Conn., USA) USP / NF It was dissolved or suspended in a polymer solution obtained by dissolving in a grade solvent (NMP, DMSO, benzyl alcohol, benzyl benzoate, ethanol). The resulting mixture was evaluated in vitro and in vivo.

Manufacturing and Quality Control The injectable gel formulation was sterilized as an aqueous solution by final filtration through a 0.22 micron filter and then aseptically dried. The sterile polymer solution and sterile GlcNAc powder were mixed by aseptic technique at the time of use. The identity, purity, potency, sterility and loading of each formulation was recorded in the production batch record. Identity, purity, potency and loading were measured using HPLC or FT-IR. Sterility was determined by a modified USP sterility test. That is, after dissolving the sample in a suitable solvent (usually DMSO), serially dilute with sterile water to a level where the solvent present is no longer bacteriostatic. The sample diluted in this way is subjected to a standard USP sterility test (USP monograph number <71>).

Release of GlcNAc from formulations incubated in phosphate buffered saline
In vitro release experiments were performed with phosphate buffered saline to evaluate the release rate under physiological conditions. Each sustained release formulation (100 μL) was added to 1.0 mL phosphate buffered saline followed by incubation at 37 ° C. At different time points, the formulation matrix was separated by pipetting away the phosphate buffered saline bath solution. After the GlcNAc derivative was generated by the method described previously (Reissig et al., J. Biol. Chem. 1955 217: 959-966), the resulting solution was analyzed by UV detection. The formulation matrix was then resuspended in 1.0 mL phosphate buffered saline and incubated under the same conditions as described above. The results are shown in FIG. 7 below.

animal
All New Zealand White rabbits (8-12 months old, weight: 3.7-4.2 kg, epiphyseal closure) were used except 3.0-3.5 kg rabbits in the Alzette pump experiment described below. Used in experiments. All experiments were conducted according to AACL guidelines and approved by the Animal Review Boards of both the University of California, San Diego and the Scripps Institute.

Anterior cruciate ligament cutting (ACLT)
Unilateral or bilateral ACLT was performed as described in each experimental set. ACLT was performed by a medial arthrotomy technique (Yoshioka et al., Osteoarthritis Cartilage 1996, 4: 87-98). After removing the patella laterally, the ACL was cut with a sharp blade. The completion of cutting was confirmed by a manual forward pull test. The knee joint was washed with sterile saline and the layers were closed with sutures. All rabbits were maintained individually with free activity. All rabbits were killed 8 weeks after the surgical procedure. Previously published data show that cartilage degeneration occurs at this point in the majority of rabbits treated with ACLT (Sah et al., J Orthop Res 1997 Mar; 15: 197-203 ).

Intramuscular injection of GlcNAc Intramuscular injection of GlcNAc was started 3 weeks a week, 1 week after the treatment, and was carried out for 7 weeks. The dose of GlcNAc per injection was 200 mg / kg. In the control group, normal saline was administered intramuscularly the same number of times.

Intra-articular injection of GlcNAc Intra-articular injection of GlcNAc started 1 week after the treatment and was carried out for 7 weeks. Rabbits were injected twice a week with 0.3 mL of GlcNAc per knee joint. The single dose of GlcNAc per injection was 80 mg. Control rabbits were injected intra-articularly with normal saline twice a week (0.3 mL per joint). The third group of rabbits received intra-articular injection of hyaluronan (0.3 mL per joint) twice weekly for 7 weeks, starting one week after ACL amputation. Synovial fluid analysis was performed in three rabbits (two in the control group and one in the hyaluronan group) that developed gross synovial effusions. The synovial fluid was culture negative in all three rabbits.

Comprehensive morphological evaluation of knee joints Overall morphological evaluation of knee joints included joint swelling evaluation, synovial fluid exudation, macroscopic articular cartilage morphology of tibial plateau and femoral condyle, and meniscus. .

  Joint swelling was assessed using the following scoring system. Grade 0-normal; Grade 1 (mild swelling)-mild inflammation and / or proliferation of the joint capsule; Grade 2 (moderate swelling)-thickening of the joint capsule and / or inflammation of the synovium; Grade 3 (severe Swelling)-sufficient inflammation of the synovium, swelling of the meniscus and ligaments (anterior or posterior cruciate ligament).

  Synovial fluid exudation was evaluated using the following scoring system. Grade 0-normal; Grade 1 (mild exudation)-more exudate than normal, but knee joint is not filled with exudate; Grade 2 (moderate exudation)-knee joint is filled with exudate However, exudate does not discharge from the joint capsule when it opens; Grade 3 (severe exudation) —The exudate causes the knee joint to expand and exudates to discharge when the joint capsule opens.

Overall Morphological Evaluation of Articular Cartilage The distal femur and proximal tibia were collected leaving a bone axis of 3.5 cm to 4 cm. The articular cartilage surface of each sample was covered with a solution of India Ink (Eberhard Faber, Lewisburg, TN) dissolved in PBS at a ratio of 1: 5. Excess ink solution was removed by gently blotting with a tissue pre-wetted with PBS. All joints were then photographed and digital images were analyzed.

  Articular cartilage was evaluated using the following scoring system. Grade 1 (intact surface)-surface is normal and does not hold India ink; Grade 2 (fine fibrosis)-India ink is held on the surface and looks like a long and light spot Grade 3 (obvious fibrosis)-velvety appearance, retains Indian ink and looks like dark black spots; Grade 4 (erosion)-loss of cartilage and underlying bone Is exposed.

Digital image The joint surface of the femoral condyle and the tibial plateau were gently sucked with a tissue and dried, and the loose tissue was removed. Each femoral axis was fixed to an optical bench. An image of the femoral condyle (resolution: 60 pixels / mm, magnification on screen: 20 ×) was obtained using a Canon EOS D30 digital camera equipped with a 100 mm macro lens at a distance of about 12 cm. A millimeter scale was placed in the photo to accurately measure the image. The magnified image was then projected onto the 3D model of the femoral condyle. The 3D surface area of the lesion was measured by plotting the edges of the lesion interactively. A digital image of the tibial joint surface was obtained as described above. The 3D projection was not used because the tibial surface was relatively flat and the 2D measurements were not significantly different from the 3D measurements.

Histological grading of the knee joint The distal thigh and proximal tibia obtained from the rabbit knee joint were fixed in 10% buffered formalin and transferred to a TBD-2 decalcifier (Thermochandon, Pittsburgh, CA). And decalcified and embedded in a paraffin block. The sagittal section of the femoral lateral condyle and medial femoral condyle and the coronal section of the tibial plateau were used for further histological analysis.

  Evaluation of sulfated glycosaminoglycan (SGAG) content was performed after staining tissue sections with Safranin O / First Green.

  SGAG content was evaluated using the following scoring system. Grade 1-Safranin O staining loss is less than 25%; Grade 2-Safranin O staining loss is 25-50%; Grade 3-Safranin O staining loss is greater than 50%.

  The following scoring system was used to assess the integrity of cartilage. Grade 1-intact cartilage surface; Grade 2-presence of fibrosis; Grade 3-full thickness cartilage defect. In addition, all tissue samples were analyzed for the presence of chondrocyte proliferation and cloning.

  Histological evaluation of the synovium was performed based on the presence of synovial proliferation and synovial neovascularization, and was separately performed for the synovium attached to the tibial plateau, the outer femoral condyle and the inner femoral condyle.

  As shown in Table 3, the microscopic evaluation of bone marrow was performed based on the presence of subchondral bone marrow hyperplasia and increased angiogenesis.

Measurement of DNA content in synovial tissue Evaluation of synovial tissue cellularity was performed by quantification of tissue DNA concentration (Amiel et al., J Orthop Res 1986; 4: 162-172). That is, the washed and freeze-dried synovial tissue was solubilized by incubation in 1N NaOH at 65 ° C. for 2 hours. A similarly prepared aliquot was reacted with 0.04% indole-HCl reagent and mixed with chloroform to remove interfering substances. The aqueous phase containing DNA was collected and the absorbance was measured at 490 nm. Calf thymus DNA was used as a standard. The results were expressed as mg DNA per mg dry tissue.

  Statistical analysis of experimental data was performed using Microsoft Excel Analysis ToolPak.

GlcNAc Alzette pump administration In this experiment, New Zealand White rabbits (3.0-3.5 kg) were used. The rabbits were randomly assigned to 5 groups (8 per group). Group A is treated with saline (negative control group), Group B is treated with 1.5 M GlcNAc, Group C is treated with 0.5 M GlcNAc, and Group D is treated with 0.15 M GlcNAc. Group E was treated with 0.05 M GlcNAc. All compounds were delivered continuously to the joint by an Alzette minipump. The delivery rate of this pump was 2.5 μL / hour. All rabbits received ACLT treatment on the right knee and GlcNAc was delivered to the right knee.

  GlcNAc administration to the right knee was carried out with an Alzette pump for 8 weeks starting immediately after ACLT treatment. The pump was changed at the end of the fourth week. The pump and delivery tube were checked twice a week to ensure that they were in place in the joint. At the end of the experiment, a photo was taken with a digital camera to show that the polyethylene tube (inner diameter: 0.58 mm) was still in the joint.

  Overall morphological changes (joint swelling, synovial fluid, etc.) of both knees were evaluated. The distal femur and proximal tibia of each of the treated and contralateral control knees were collected. The occurrence, location and severity of lesions on the sample surface were judged by setting criteria under an optical microscope. The femoral condyle and tibial plateau stained with India ink were also photographed with a digital camera. The surface area of the stained lesion on the digital image was quantified by image analysis software and compared between groups.

  The efficacy of GlcNAc intramuscular injection was evaluated in 6 rabbits that received bilateral ACLT, and compared to 6 rabbits that received bilateral ACLT but were injected intramuscularly with saline alone. From a comprehensive morphological analysis of the tibial plateau and femoral condyle, there was no statistically significant difference in the degree of cartilage damage between the GlcNAc-treated and control populations. Figures 1a and 1b are plots of the overall morphological evaluation of these groups. FIG. 1a shows the evaluation of the femoral condyle and FIG. 1b shows the evaluation of the tibial plateau.

  On the other hand, it can be seen that intraarticular injection of GlcNAc improves tibial plateau and femoral condyle damage. Intra-articular injections of GlcNAc (treatment group, n = 7) or physiological saline intra-articular injection (control group, n = 7) were given twice a week for a total of 7 weeks to the rabbits subjected to bilateral ACLT. As shown in FIG. 2, from the comprehensive morphological analysis of the femoral condyle, the treatment group showed a tendency toward improvement of cartilage damage (improvement of lesions) compared to the control group. Furthermore, as shown in FIG. 3, the morphological analysis of the tibial plateau revealed a significant chondroprotective activity of GlcNAc. That is, cartilage lesions developed in 6 of 7 rabbits in the control group, whereas only 7 rabbits developed cartilage lesions in 7 rabbits in the treatment group (p <0. 003). Figures 4 and 5 show that intra-articular administration of GlcNAc has no significant effect on joint swelling, synovial effusion and DNA content in synovial tissue.

  Thus, in the case of GlcNAc intramuscular injection, although cartilage protective action is not shown, a tendency to suppress synovitis is observed. On the other hand, in the case of intra-articular administration of GlcNAc, significant suppression of cartilage degeneration is seen at the macroscopic level and the microscopic level.

  Next, a comparative experiment between GlcNAc intra-articular administration and hyaluronan intra-articular administration was performed. As mentioned above, hyaluronan formulations are commonly used as viscosupplementation for the treatment of knee osteoarthritis. In this experiment, no significant difference was found between the GlcNAc group and the hyaluronan group (n = 7) from the comprehensive morphological analysis of the femoral condyle (FIG. 2). However, as can be seen from FIG. 3, GlcNAc exhibits significantly higher (p <0.01) cartilage protective activity than hyaluronan. In addition, the surface area of the cartilage lesion is significantly reduced in the GlcNAc group compared to the hyaluronan group (FIGS. 6a and 6b). There is no significant difference in synovial fluid exudation between the GlcNAc group and the hyaluronan group (FIG. 4). However, DNA content evaluation shows that synovial hyperplasia and cellularity are significantly suppressed (p <0.05) in the GlcNAc group compared to the hyaluronan group (FIG. 5).

  Histological analysis of experimental rabbits treated with GlcNAc or saline was performed and the results obtained from each population were compared. As can be seen from Table 1 below, similar SGAG loss is seen in the medial femoral condyle in both the GlcNAc treated and control populations. However, from the analysis and comparison of SGAG in the tibial plateau, it can be seen that there is a trend toward improvement in the GlcNAc group and that SGAG loss is significantly suppressed in the lateral femoral condyles in the GlcNAc group. In addition, as a result of examining cartilage integrity for these groups, the GlcNAc group shows significant cartilage protective activity on the tibial plateau and femoral lateral condyle as compared to the control group.

  Histological evaluation of synovitis shows that GlcNAc inhibits synovial proliferation (Table 2). The effect of GlcNAc on synovial neovascularization varies from site to site, but significant improvement is shown by GlcNAc near the femoral medial condyle (Table 2). From examination of subchondral bone marrow, it can be seen that GlcNAc treatment suppresses capillary dilution and hyperplasia that cause subchondral bone edema (Table 3).

  In summary, the histological analysis of cartilage, synovium and subchondral bone marrow / edema shows the cartilage protective and anti-inflammatory effects of intra-articularly administered GlcNAc.

  From these results, it can be seen that intraarticular administration of GlcNAc significantly and unexpectedly suppressed cartilage degeneration as measured by macroscopic and microscopic criteria. In addition, the greatest cartilage protective activity was observed in the tibial plateau, followed by the outer femoral condyle and the inner femoral condyle. From this result, the joint site with lower severity of cartilage destruction is more sensitive to the therapeutic action of GlcNAc. It was strongly shown to be expensive. Finally, intra-articular GlcNAc administration unexpectedly reduced the severity of synovitis and improved subchondral bone marrow hyperplasia. Furthermore, delivery of GlcNAc to the joint can be performed continuously for more than 3 weeks after a single dose (intra-joint) as required by the sustained release and alzette pump methods described above.

  Upon intra-articular administration of GlcNAc, the greatest cartilage protective activity was seen on the tibial plateau and followed by the outer femoral condyle and the medial femoral condyle. It was found that GlcNAc administration is more responsive than the site of higher severity of cartilage damage. On the other hand, when GlcNAc was administered intramuscularly to rabbits with experimental OA, the cartilage protective effect was not shown, but significant suppression of synovitis (ie, anti-inflammatory activity) was shown. Intra-articular administration of GlcNAc was much more potent than intramuscular administration. Rabbits treated with intra-articular GlcNAc administration showed a significant delay in cartilage degeneration at both macroscopic and microscopic levels. Finally, intra-articular GlcNAc administration is superior to visco-supplementation therapy using hyaluronan in terms of its cartilage protective effect.

  In summary, intraarticular therapy with GlcNAc in experimental OA rabbits unexpectedly suppresses cartilage degeneration, macroscopically reduces rabbit lesion size, significantly reduces synovitis, and subchondral bone edema Bone marrow hyperplasia was suppressed.

Pharmaceutical sugars and amino sugars (most preferably GlcNAc) as active ingredients for administration , once isolated, are pharmaceutically acceptable preparations such as Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing, Easton, Pa. 1990) (incorporated herein as part of this specification) and can be incorporated into a specific treatment of a disease or disorder if it has little or no effect on healthy tissue. Can be used. The preparation of a pharmacological composition comprising an active ingredient dissolved or dispersed need not be limited based on the formulation. Such compositions can be prepared as injectable solutions or suspensions. However, it is also possible to prepare a solid suitable for dissolution or resuspension in a liquid before use. The preparation can also be emulsified.

  In a preferred embodiment, a label (or equivalent label in other countries) is attached that indicates that the composition is approved by the US Food and Drug Administration (FDA) for treating the diseases and disorders described herein. The composition is contained in a sealed container. Such containers provide a therapeutically effective amount of the active ingredient to be administered to the host.

  The specific amino sugar that affects the target disorder can be administered to a mammal alone or as a pharmaceutical composition in which the amino sugar is mixed with an appropriate carrier or excipient. In treating a mammal exhibiting a target disorder, a therapeutically effective amount of an agent (eg, GlcNAc) is administered. The active ingredient can be mixed with an excipient that is pharmaceutically acceptable and compatible with the active ingredient in an amount suitable for use in the therapeutic methods described herein.

  Pharmaceutically acceptable salts can be prepared by standard techniques. For example, first, the free base type compound is dissolved in a suitable solvent (for example, an aqueous solution or a water-alcohol solution) containing a suitable acid. The salt is then isolated by evaporating the solution. In another example, the salt is prepared by reacting the free base with an acid in an organic solvent.

  Carriers and excipients can be used to facilitate administration of the compound (eg, increase the solubility of the compound). Examples of carriers and excipients include calcium carbonate and calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oil, polyethylene glycol, water, physiological saline, dextrose, glycerol, ethanol, and physiologically compatible solvents.

  The compositions of the present invention can include pharmaceutically acceptable salts of the active ingredients. Pharmaceutically acceptable salts include acid additions formed from free amino groups of amino sugars and inorganic acids (eg, hydrochloric acid, phosphoric acid, sulfuric acid, etc.) or organic acids (acetic acid, tartaric acid, mandelic acid, etc.). Salt. In addition, a free carboxyl group of an amino sugar and an inorganic base (for example, sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, ferric hydroxide, etc.) or an organic base (for example, isopropylamine, trimethylamine, 2 -Aminoethanol, histidine, procaine, etc.) can also form salts.

  The toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures and laboratory animals, such as LD50 (amount that causes 50% of the population to die) and ED50 (50% of the population). A therapeutically effective amount). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50 / ED50. Compounds with large therapeutic indices are preferred. Data obtained from such cell culture assays and animal experiments can be used to set a dose range for human use. The dosage of such compounds lies preferably within a blood concentration range that includes the ED50 with little or no toxicity. The dose of the compound may vary within this range depending on the dosage form used and the route of administration.

  For any amino sugar compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. For example, a test compound that achieves the IC50 (ie, half-maximal disruption of a protein complex or half-maximal inhibition of the cellular level and / or activity of a complex component as required in cell culture. Doses can be set in animal models to achieve a circulating plasma concentration range including Such information can be used to more accurately determine useful doses in humans. The plasma concentration can be measured, for example, by HPLC.

  Another preferred embodiment of the present invention relates to an improved formulation of the active ingredient GlcNAc. Encapsulating or taking GlcNAc in liposomes or other uptake agents changes its pharmacodynamic profile upon intra-articular injection. Preferably, GlcNAc is incorporated into the matrix. More preferably, the GlcNAc is incorporated within a matrix selected from the group consisting of particles, implants and gels.

  The exact formulation, route of administration, and dose can be selected by an individual physician in view of mammalian disorders (eg, Fingl et al., The Pharmacological Basis of Therapeutics, 1975, Ch. 1, p. 1 reference). Of course, the attending physician will know how and when to terminate, interrupt, or adjust administration due to toxicity or organ dysfunction. Conversely, the attending physician will also know to adjust the level of treatment higher if the clinical response is not sufficient (excludes toxicity). The amount of dosage in the treatment of the target disorder varies depending on the severity of the disorder to be treated and the route of administration. The severity of the disorder can be partially evaluated by, for example, a standard prognostic evaluation method. Also, the dose, and possibly the frequency of administration, will vary depending on the age, weight and response of the individual mammal. Anything comparable to the above program can be used in veterinary medicine.

  Depending on the particular disorder being treated, the agents described above can be formulated and administered systemically or locally. Formulation and administration techniques can be found in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing, Easton, Pa. (1990), which is incorporated herein by reference. To do.

  In the case of injection, the agent of the present invention can be formulated into an aqueous solution, preferably a physiologically compatible buffer (for example, Hank's solution, Ringer's solution, physiological saline buffer).

  It is within the scope of this invention to prepare a compound disclosed herein in a dosage form suitable for systemic administration using a pharmaceutically acceptable carrier. By appropriate selection of the carrier and appropriate manufacture, the compositions of the present invention, particularly those formulated as solutions, can be administered parenterally, eg, intravenously injected.

  Pharmaceutical compositions suitable for use in the present invention include compositions containing an active ingredient in an effective amount to achieve its intended purpose. Effective amounts can be determined within the capabilities of those skilled in the art, especially in light of the detailed disclosure herein. Such pharmaceutical compositions contain appropriate pharmaceutically acceptable carriers that contain, in addition to the active ingredient, adjuvants and excipients that facilitate the processing of the active compound into pharmaceutically acceptable formulations. It can be included. The pharmaceutical composition of the present invention can be produced by a method known per se, for example, a conventional mixing process, dissolution process, granulation process, dragee manufacturing process, levitating process, emulsification process, encapsulation process, uptake process, lyophilization. It can be manufactured by a process.

  Pharmaceutical formulations for parenteral administration include aqueous solutions of water soluble active compounds. In addition, suspensions of the active compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate and triglycerides, and liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. If necessary, a high-concentration solution can be prepared by adding an appropriate stabilizer or an agent for increasing the solubility of the compound to the suspension.

  Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, depending on the needs, such as gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions, and suitable organic solvents or A solvent mixture can be included. Dyes and pigments can be added to the tablets and dragee coatings for identification and to characterize different combinations of active compound administration.

  The present invention described in the present description can be appropriately implemented in the absence of elements or limitations not specifically disclosed in the present specification. For example, terms such as “comprising”, “including”, and “containing” should be interpreted broadly without limitation. In addition, the terms and expressions described in this specification are used as terms for explanation rather than as terms for limitation, and in using such terms and expressions, any of the features described It will be understood that it is not intended to exclude equivalents or any part thereof, and that various modifications are possible within the scope of the claimed invention. Thus, it should be understood that although the present invention has been specifically disclosed by way of preferred embodiments and optional features, modifications and variations of the invention disclosed herein may be rearranged by those skilled in the art. Such changes and modifications are considered to be within the scope of the invention disclosed herein. The present invention has been described broadly and comprehensively herein. Each subgeneric species within the scope of this comprehensive disclosure also forms part of the invention. This includes a comprehensive description of each invention, subject to the removal of any content from this genus or a negative limitation, but it does not matter whether the omitted content was specifically there. . Furthermore, if the features and aspects of the invention are described in the form of a Markush group, those skilled in the art will also describe the invention as an individual element of the Markush group or a subgroup of elements thereof. Will understand.

  It will be understood that the above description is illustrative and not restrictive. Many embodiments will be apparent to those of skill in the art upon reviewing the above description. Accordingly, the scope of the invention should not be determined by the foregoing description, but instead should be determined by the appended claims and the full scope of equivalents based on those claims. The disclosures of all papers and references (including patent publications) are hereby incorporated by reference as part of this specification.

FIG. 1A shows the overall morphological grading of the femoral condyle in rabbits subjected to bilateral anterior cruciate ligament amputation (ACLT) and treated by intramuscular injection of GlcNAc or normal saline. FIG. 1B shows the overall morphological grading of the tibial plateau in rabbits subjected to bilateral anterior cruciate ligament (ACL) amputation and treated by intramuscular injection of GlcNAc or normal saline. FIG. 2 shows the overall morphological grading of the femoral condyle in rabbits subjected to unilateral ACL amputation and treated by intra-articular injection of GlcNAc, sodium hyaluronate or saline. FIG. 3 shows the overall morphological grading of the tibial plateau in rabbits subjected to unilateral ACL cutting and treated by intra-articular injection of GlcNAc, sodium hyaluronate or saline. FIG. 4 shows a comprehensive morphological evaluation of joint swelling in rabbits subjected to unilateral ACL cutting and treated by intra-articular injection of GlcNAc, sodium hyaluronate or saline. FIG. 5 shows the DNA content in synovial tissue obtained from rabbits subjected to unilateral ACL cutting and treated by intra-articular injection of GlcNAc, sodium hyaluronate or saline. FIG. 6A shows a digital image analysis of lesion size in the femoral condyle obtained from rabbits subjected to unilateral ACL amputation and treated by intra-articular injection of GlcNAc or sodium hyaluronate. FIG. 6B shows digital image analysis of lesion size in the tibial plateau obtained from rabbits subjected to unilateral ACL cutting and treated by intra-articular injection of GlcNAc or sodium hyaluronate. FIG. 7 shows the time-dependent in vitro release of GlcNAc incorporated into an injectable polymer formulation according to one embodiment of the present invention.

Claims (33)

  A method for treating a disease in a mammal, wherein the disease is selected from the group consisting of synovitis, subchondral bone edema and cartilage degeneration, wherein the treatment administers a therapeutically effective amount of an amino sugar to the mammal. A method involving that.   The method of claim 1, wherein the amino sugar is selected from the group consisting of N-acetylglucosamine, glucosamine, galactosamine, N-acetylgalactosamine, iminocyclitol, and pharmaceutically acceptable salts thereof.   The method of claim 1, wherein the amino sugar is incorporated into a matrix.   4. The method of claim 3, wherein the matrix is selected from the group consisting of particles, implants and gels.   5. A method according to claim 4, wherein the particles are liposomes, nanospheres, microspheres or suspensions.   The method of claim 4, wherein the implant is a polymer, pump or device.   The method of claim 4, wherein the gel is an in situ implant-forming gel, a semi-solid gel, a hydrogel, or a thermosensitive gel.   An injectable formulation for intra-articular treatment of diseases associated with joint disorders, comprising an amino sugar incorporated into the matrix, wherein the matrix is a particle, implant or gel.   A method of treating a disease associated with joint disorders comprising administering a therapeutically effective amount of N-acetylglucosamine as a controlled release formulation.   10. The method of claim 9, wherein N-acetylglucosamine is administered by intramuscular or intraarticular injection.   10. The method of claim 9, wherein N-acetylglucosamine is administered by subcutaneous injection or infusion.   10. The method of claim 9, wherein the disease associated with joint damage is selected from the group consisting of synovitis, subchondral bone edema and cartilage degeneration.   13. The method of claim 12, wherein the disease is synovitis.   13. The method of claim 12, wherein the disease is subchondral bone edema.   13. The method of claim 12, wherein the disease is cartilage degeneration.   The method of claim 9, wherein the joint disorder is not osteoarthritis.   10. The method of claim 9, wherein the joint disorder is not rheumatoid arthritis. a. Diagnosing pathological markers associated with joint disorders;
b. Administering an amino sugar as a therapeutically effective formulation.   19. The method of claim 18, wherein the pathological marker is selected from the group consisting of synovitis, subchondral bone edema and cartilage degeneration.   20. The method of claim 19, wherein the pathological marker is synovitis.   20. The method of claim 19, wherein the pathological marker is subchondral bone edema.   20. The method of claim 19, wherein the pathological marker is cartilage degeneration.   The method of claim 18, wherein the joint disorder is not osteoarthritis.   19. The method of claim 18, wherein the joint disorder is not rheumatoid arthritis.   19. The method of claim 18, wherein administering the amino sugar is performed by a route of administration selected from the group consisting of intra-articular, intramuscular, infusion pump and subcutaneous.   26. The method of claim 25, wherein the route of administration is intra-articular.   The amino sugar is selected from the group consisting of N-acetylglucosamine, glucosamine, galactosamine, N-acetylgalactosamine, iminocyclitol, combination therapies thereof, pharmaceutically acceptable salts thereof, and injectable preparations. The method of claim 18, wherein the method is selected.   28. The method of claim 27, wherein the amino sugar is N-acetylglucosamine.   28. The method of claim 27, wherein the injectable formulation is selected from the group consisting of matrix particles, matrix gels and controlled release formulations.   28. The method of claim 27, wherein the combination therapy includes a combination of the amino sugar and a compound selected from the group consisting of an anti-inflammatory drug and a hexosaminidase inhibitor.   A method for preventing cartilage degeneration, comprising administering a therapeutically effective amount of N-acetylglucosamine as a controlled release preparation to a mammal having a low severity of cartilage degeneration.   32. The method of claim 31, wherein the N-acetylglucosamine is administered by intramuscular or intraarticular injection.   The method of claim 21, wherein the N-acetylglucosamine is administered by subcutaneous injection or infusion pump.

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