STABILIZED POLYESACARIDE COMPOSITIONS FOR THE TREATMENT OF A JOINT AND KIT CONDITION

专利摘要:
compositions and methods for stabilized polysaccharide formulations. the present invention relates to compositions and methods for treating joint conditions, such as osteoarthritis and / or the pain associated with it. the compositions and methods use a first component, namely hyaluronic acid (ha), in combination with at least one stabilizer. the composition may include a stabilizer that increases the stability and the life of the ha. in another embodiment, the compositions and methods may also include an add component, such as one or more glycosaminoglycans (gag) or precursors of the gag. examples of gags or gag precursors may include chondroitin sulfate (cs), dermatan sulfate, heparin, heparan sulfate, keratan sulfate and glycosamine (gicn).
公开号:BR102012016298B1
申请号:R102012016298-9
申请日:2012-06-29
公开日:2020-09-24
发明作者:Dongling Su；Julia Hwang；Brooks J. Story
申请人:Depuy Mitek, Inc.；
IPC主号:

专利说明:

[0001] [001] The present invention relates in general to compositions and methods for treating joints. BACKGROUND OF THE INVENTION
[0002] [002] Osteoarthritis ("OA"), the most common form of arthritis, is a type of arthritis that is characterized by degenerative (gradual deterioration of the joint) or abnormal changes in the bone, cartilage and synovium of the joints. OA is characterized by progressive wear of the surfaces of opposite joints, sometimes accompanied by inflammation that results in pain, swelling and stiffness of the joint. OA can occur in one or more joints, followed by trauma to the joint, followed by an infection of the joint or simply as a result of aging. In addition, there is emerging evidence that abnormal anatomy, genetics and obesity can contribute to the early development of OA.
[0003] [003] Treatment of OA in general involves a combination of exercise, physical therapy, lifestyle modification and pain relievers. Acetaminophen is typically the first-line treatment for OA. For mild to moderate symptoms, the efficacy is similar to that of non-steroidal anti-inflammatory drugs ("NSAIDs"), such as ibuprofen. For more severe symptoms, NSAIDs can be more effective. However, although NSAIDs are more effective in severe cases, they are associated with greater side effects such as gastrointestinal bleeding and kidney complications. Another class of NSAIDs, selective COX-2 inhibitors (like Celecoxib), are as effective as NSAIDs, but are no longer safe in terms of side effects. There are several NSAIDs available for topical use, including diclofenac. Typically, they have fewer systemic side effects than oral administration and at least some therapeutic effects. Although opioid analgesics, such as morphine and fentanyl, improve pain, this benefit is supplanted by frequent adverse events and, therefore, they are not routinely used. Intra-articular steroid injections are also used to treat OA, and they are very effective in providing pain relief, specifically in patients who have inflammatory OA elements. However, the duration of pain relief is limited to 4 to 6 and there are adverse effects that can include damage to side cartilage. If the pain becomes debilitating, joint replacement surgery can be used to optimize mobility and quality of life. There is no proven treatment to slow or reverse the disease.
[0004] [004] For patients who cannot achieve adequate pain relief using simple pain relievers, such as acetaminophen or exercise and physical therapy, intrarticular injections of hyaluronic acid (HA) provide another treatment option to treat symptomatic pain and can delay need for total joint replacement surgery. It is known that the concentration and molecular weight of native HA is deficient in individuals suffering from OA and, therefore, injections into the joint of exogenous HA are believed to restore these molecules and restore the viscoelastic properties of synovial fluid. This is the property that is responsible for lubricating and cushioning the joints. There is also evidence that HA has biological activity through binding to cell surface receptors and may play a role in mitigating cartilage inflammation and degradation. Regardless of the mechanism of action, pain relief is seen for about six months after the course of treatment. A course of treatment for HA products in the North American market can range from a single injection to others that require 3 to 5 injections per week to achieve this pain-relieving durability.
[0005] [005] Currently, hyaluronic acid ("HA") formulations on the market in the United States are marketed as liquid HA solutions ready for use in pre-filled syringes. They can be stored at room temperature, and typically have a shelf life of two years. Although low to moderate molecular weight HA can be effective, high molecular weight HA formulas can provide additional benefits, specifically at higher concentrations of HA. However, HA in solution is known to degrade over time at room temperature, as measured by a reduction in the molecular weight of HA, which could impact its effectiveness as an OA therapy.
[0006] [006] There remains a need for improved methods and compositions for treating joints, and, in particular, for improved methods and compositions for treating joints using high molecular weight HA, alone or combined with additional components, to treat the stability and service life problems associated with current treatments. SUMMARY OF THE INVENTION
[0007] [007] Compositions and methods are provided for the treatment of joint conditions, such as osteoarthritis and / or the pain associated with it. The methods and compositions presented in the present invention are generally directed at treating joints using a first component, namely, hyaluronic acid ("HA"), together with at least one stabilizer.
[0008] [008] In one aspect, formulations or compositions are presented for the treatment of an articulation condition that comprises a formulation. The formulation or composition can be in liquid form. The formulation can also be stable at room temperature. In addition, the formulation may include a hyaluronic acid (HA) solution. The HA formulation can be a high molecular weight HA. The molecular weight can be, for example, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000, 5100, 5200, 5300, 5400, 5500, 5600, 5700, 5800, 5900, 6000 kDa or more, or any derivable range therein. In exemplary modalities, HA has a molecular weight in the range of about 1 MDa to 6 MDa. In another exemplary embodiment, HA has a molecular weight greater than 1 MDa.
[0009] [009] In addition, the HA formulation can be present in particular concentrations in liquid, solid or lyophilized form. In one embodiment, HA is present in a liquid concentration of at least about 1 mg / mL. In another exemplary embodiment, HA has a liquid concentration of at least about 5 mg / ml, and more preferably of at least about 7 mg / ml, and more preferably of at least about 10 mg / ml, and more preferably at least about 15 mg / ml, and in some embodiments the concentration can be at least about 25 mg / ml. In another embodiment, HA can have a concentration in the range of about 15 mg / ml to about 25 mg / ml.
[0010] [0010] In another aspect, the formulation or composition includes at least one additional component. The additional component added to the formulation or composition can be, for example, amino acids, amino sugars, sugar alcohols, proteins, saccharides, disaccharides, oligosaccharides, polysaccharides, nucleic acids, buffers, surfactants, lipids, liposomes, other excipients and mixtures thereof . Other useful components may include steroids, anti-inflammatory agents, non-steroidal anti-inflammatory agents, pain relievers, cells, antibiotics, microbicidal agents, anti-inflammatory agents, growth factors, growth factor fragments, small molecule wound healing stimulants, hormones , cytokines, peptides, antibodies, enzymes, isolated cells, platelets, immunosuppressants, nucleic acids, cell types, viruses, viral particles, essential nutrients, minerals, metals or vitamins, and combinations thereof. In addition, the formulation or composition can include a diluent, such as water, saline or a buffer.
[0011] [0011] In an exemplary embodiment, the formulation or composition includes at least one stabilizer or stabilizing excipient, such as tocopherol, tocopherol derivatives, mannitol, glucose, sucrose and trehalose. The stabilizers can be present in the range of about 0.1 to 70% by weight; from about 0.1 to 50% by weight, from about 0.1 to 20% by weight; or about 0.5 to 20% by weight. Alternatively, the excipient can be present in a concentration in the range of about 1 mg / ml to about 700 mg / ml, and more preferably in the range of about 1 mg / ml to about 500 mg / ml, and with more preferably in the range of about 1 mg / ml to about 200 mg / ml, and most preferably in the range of about 5 mg / ml to about 200 mg / ml.
[0012] [0012] In another aspect, the formulation or composition includes at least one additional component, such as a glycosaminoglycan (GAG), including chondroitin sulfate ("CS") GAGs, dermatan sulfate, heparin, heparan sulfate and sulfate keratan, or a GAG precursor such as glucosamine ("GlcN"). In one embodiment, a GAG or GAG precursor may be present in the composition at a concentration of at least about 0.1 mg / ml, and more preferably at least about 2 mg / ml, and more preferably at least less about 5 mg / ml, and more preferably at least about 7 mg / ml, and more preferably at least about 20 mg / ml. In another modality, GAGs can have several molecular weights, however in certain exemplifying modalities the molecular weight is in the range of about 5 to 1,000 KDa, more preferably in the range of about 6 to 500 KDa, more preferably in the range of about 7 to 300 KDa, more preferably in the range of about 8 to 200 KDa, most preferably in the range of about 9 to 100 KDa and, most preferably, in the range of about 10 to 80 KDa. In other embodiments, the molecular weight of a GAG fragment can be below about 5 KDa and, more preferably, below about 3 KDa. In another embodiment, the GAG precursor may have a molecular weight of about 180 Da In other embodiments, the GAG precursor may have a molecular weight of at least about 100 Da. In an exemplary embodiment, the GAG precursor may have a molecular weight in the range of about 100 Da to about 250 Da.
[0013] [0013] Furthermore, the formulation or composition may have a weight to HA ratio in the range of about 1: 0.001 to about 1: 100, and more preferably in a ratio of about 1: 0.0125 to about 1:10, about 1: 0.00125 to about 1: 100, and more preferably in a ratio in the range of about 1: 0.01 to about 1:10. Alternatively, the formulation or composition can include about 1% to about 75% or more, by weight, of any of the individual components, for example, HA and a stabilizer.
[0014] [0014] One or more aspects may include a kit and methods of using the kit. The kit may include a solution of high molecular weight hyaluronic acid (HA) and at least one stabilizer. The kit can be stored at room temperature. In addition, the kit can include a syringe containing the HA solution and the stabilizer in a single chamber. Another embodiment may include a single chamber with the HA solution and stabilizer and a separate chamber with an additional component, such as a GAG or GAG precursor. For example, a glycosaminoglycan (GAG) may be present in the kit in a chamber separate from the HA / stabilizer solution. The additional component may be present as a liquid, solid or lyophilized. The kit can also include a diluent, such as water, saline and a buffer, to solubilize one of the components, the additional component.
[0015] [0015] In an additional aspect, a method for treating the joints is presented that includes administering a liquid formulation comprising high molecular weight hyaluronic acid to a needy individual. The method may include high molecular weight hyaluronic acid in the presence of a stabilizer. In addition, high molecular weight hyaluronic acid can be stored in a liquid formulation at room temperature before administration. The method may also include administration of the HA solution present in the kit or present in the high molecular weight HA formulation or composition. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] [0016] The attached drawings have been included in the present invention so that the characteristics, advantages and objects mentioned above will become clear and can be understood in detail. These drawings form a part of the specification. It should be noted, however, that the attached drawings illustrate exemplary modalities and should not be considered as limiting the scope.
[0017] [0017] Figure 1 is a schematic view of an embodiment of a mixing and release system for use with the compositions and methods of the present invention;
[0018] [0018] figure 2 is a perspective view of another embodiment of a mixing and release system for use with the compositions and methods of the present invention;
[0019] [0019] Figure 3 shows the degradation of HA, as determined by size exclusion chromatography-high performance liquid chromatography (SEC-HPLC) when in a saline solution of HA and CS stored at 5 ° C and 40 ° C / 75% relative humidity for 5 months;
[0020] [0020] Figure 4 shows the stabilizing effect that trehalose has on HA when in solution with CS stored at 5 ° C and 40 ° C / 75% relative humidity for 5 months as determined by SEC-HPLC;
[0021] [0021] Figure 5 is a graph comparing the widths of the cartilage lesion in a tear model of the median rat meniscus (MMT) that received intrarticular trehalose injections with those that did not receive treatment;
[0022] [0022] figure 6 is a graph that compares the ratio of the depth of the cartilage lesion in the rat MMT model that received intrarticular trehalose injections with those that did not receive treatment;
[0023] [0023] figure 7 is a graph comparing the collagen degradation in the rat MMT model that received intrarticular trehalose injections with those that did not receive treatment;
[0024] [0024] figure 8 shows the effect that trehalose had on the degeneration of the cartilage of the median tibial plateau; and
[0025] [0025] Figure 9 shows the effect that trehalose had on cartilage degradation in a rabbit anterior cruciate ligament (ACLT) model of osteoarthritis. DETAILED DESCRIPTION
[0026] [0026] In general, the present invention features compositions and methods for the treatment of joint conditions, such as osteoarthritis and / or the pain associated with it. The compositions and methods use a first component, namely, high molecular weight hyaluronic acid ("HA"), alone or in combination with an additional component, combined with a stabilizer to prolong the service life. In an exemplary embodiment, the composition includes a stabilizer or stabilizing excipient to stabilize HA and, if present, a precursor to GAG or GAG. Hyaluronic acid
[0027] [0027] Hyaluronic acid (HA) can have several formulations and can be supplied in various concentrations and molecular weights. The terms "hyaluronic acid", "hyaluronan", "hyaluronate" and "HA" are used interchangeably in the present invention to refer to hyaluronic acids or salts of hyaluronic acid, such as sodium, potassium, magnesium and calcium salts, among others. These terms are also intended to include not only pure hyaluronic acid solutions, but also hyaluronic acid with other trace elements or in various compositions with other elements. The terms "hyaluronic acid", "hyaluronan" and "HA" include chemical or polymeric or cross-linked derivatives of HA. Examples of chemical modifications that can be made to HA include any reaction of an agent with the four reactive groups of HA, namely the acetamido, carboxyl, hydroxyl and reducing end. The HA used in the present application aims to include natural formulations (isolated from animal tissue) or synthetic formulations (derived from bacterial fermentation) or combinations thereof. HA can be supplied in liquid form or in solid formulations that are reconstituted with diluents to achieve an appropriate concentration.
[0028] [0028] HA is a glycosaminoglycan (GAG), and in particular, HA is an unbranched polysaccharide composed of alternating glucuronic acid and N-acetyl glycosamine units. In liquid form, HA has viscoelastic properties. HA is also found in the extracellular matrix of cartilage as an important structural component of agrecane, which makes up the proteoglycan complex. The main function of the proteoglycan complex is to retain water in the cartilage matrix, conferring its characteristic of turgidity and mechanical resilience.
[0029] [0029] HA not only helps maintain the healthy mechanical properties of the cartilage that cushions the joints, but it is also a major component of synovial fluid.
[0030] [0030] Abnormalities of HA are a thread in connective tissue disorders. HA can therefore be used to prevent, treat or assist in the surgical repair of connective tissue disorders.
[0031] [0031] HA can be used in the compositions and methods of the present invention in various molecular weights. Since HA is a polymeric molecule, the HA component can exhibit a molecular weight range, and almost any HA modal molecular weight formulation medium can be used in the compositions and methods of the present invention, including low molecular weight hyaluronan ( "LWM") (about 500 to 700 kilodaltons (kDa), medium molecular weight hyaluronan ("MMW") (700 to 1000 KDa), and high molecular weight hyaluronan ("HMW") (1.0 to 6, 0 million daltons (MDa). In certain exemplary embodiments, HA has a molecular weight of at least about 700 KDa, and in certain embodiments, HA is a high molecular weight HA ("HWM") that has a molecular weight of at least about 1 MDa. The molecular weight can be, for example, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000, 5100, 5200, 5300, 5400, 5500, 5600, 5700, 5800, 5900, 6000 kDa or more, or any derivable range therein. It is expected that chemically modified HAs could have very different molecular weights than those described above. Crosslinked HA can similarly have a molecular weight that is much higher than that noted above. Regardless, these materials are also applicable in this invention.
[0032] [0032] HA can be represented in solid, lyophilized or liquid form. When in liquid form, solvents can be used to solubilize HA. Solvents may include, but are not limited to, water, saline or other salt solutions, buffer solutions such as phosphate buffered saline, histidine, lactate, succinate, glycine and glutamate, dextrose, glycerol, as well as combinations thereof.
[0033] [0033] The concentration of HA present in the formulation can also vary, however in an exemplary embodiment HA is supplied in a pharmaceutically effective amount. In one embodiment, HA has a concentration of at least about 1 mg / mL. In an exemplary embodiment, HA has a concentration of at least about 5 mg / ml, and more preferably at least about 7 mg / ml, and most preferably at least about 10 mg / ml, and with more preferably at least about 15 mg / ml, and in some embodiments the concentration can be at least about 25 mg / ml. In another embodiment, HA can have a concentration in the range of about 15 mg / ml to about 25 mg / ml. Suitable concentrations of HA include about 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml ml, 10 mg / ml, 11 mg / mg, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml ml, 20 mg / ml, 21 mg / ml, 22 mg / ml, 23 mg / ml, 24 mg / ml, 25 mg / ml, 26 mg / ml, 27 mg / ml, 28 mg / ml, 29 mg / ml ml, 30 mg / ml, 31 mg / ml, 32 mg / ml, 33 mg / ml, 34 mg / ml, 35 mg / ml, 36 mg / ml, 37 mg / ml, 38 mg / ml, 39 mg / ml ml, 40 mg / ml, 41 mg / ml, 42 mg / ml, 43 mg / ml, 44 mg / ml, 45 mg / ml, 46 mg / ml, 47 mg / ml, 48 mg / ml, 49 mg / ml ml, 50 mg / ml, 51 mg / ml, 52 mg / ml, 53 mg / ml, 54 mg / ml, 55 mg / ml, 56 mg / ml, 57 mg / ml, 58 mg / ml, 59 mg / ml L, 60 mg / mL or more or any derivable range therein.
[0034] [0034] In one embodiment, the first component comprises an HA having a high molecular weight (1 to 6 MDa) having a concentration in the range of about 5 to 40 mg / ml. One such product is Orthovisc® produced by Anika Therapeutics, Inc. of Bedford, MA. Orthovisc® is a sterile, non-pyrogenic, clear and viscoelastic solution of hyaluronan. Orthovisc® consists of natural ultra high purity hyaluronan (1.0 to 2.9 MDa) dissolved in physiological saline and with nominal concentrations between 12.5 and 17.5 mg / mL. Orthovisc® is isolated through bacterial fermentation. The person skilled in the art will recognize that there are companies like Shiseido and Lifecore that can produce high molecular weight HA through a bacterial fermentation process. Another example of an HA product available in the United States with these features is Euflexxa®. Stabilizers
[0035] [0035] HA can be combined with at least one stabilizer. Stabilizers can be used in the methods and compositions of the present invention. Stabilizers can be sugars or derivatives, such as saccharides, disaccharides, modified saccharides, sugar alcohols or polysaccharides. In an exemplary embodiment, the stabilizer can be tocopherol, tocopherol derivatives, glucose, mannitol, sucrose and / or trehalose.
[0036] [0036] Stabilizers or common stabilizing excipients used in the pharmaceutical industry are saccharides, disaccharides, modified saccharides, sugar alcohols and polysaccharides. Some of these molecules are commonly used as excipients because they can also serve as a sweetening agent when the drug is in the form of a pill for oral delivery. According to U.S. Patent No. 7,351,798, hyaluronic acid and glycosaminoglycans can also be excipients. These polysaccharides have long been believed to be stable molecules. In fact, all HA materials that are marketed in the United States are stored at room temperature. Some of these HA products are in buffered solutions or in saline solutions, but none have any other excipients included. These HA products are approved in the United States as viscosupplementation devices and are injected into the joint to relieve the pain that results from osteoarthritis. In addition, a product on the market called Viscoat (sold by Alcon) is a formulation that incorporates hyaluronic acid and chondroitin sulfate in a buffered solution and does not contain any other excipients. This product is used in ophthalmic surgery during the implantation of intraocular lenses. It is known that storage of HA at room temperature results in a gradual loss of stability and degradation to reduce molecular weights, specifically in the presence of other components, such as chondroitin sulfate, hence the requirement for cold storage for Viscoat
[0037] [0037] Tocopherol belongs to a class of chemical compounds that cover mono, di and trimethylcotols. Many of the tocopherols show vitamin E activity. Beta, gamma and delta are stereoisomers of alpha-tocopherol. Tocopherol esters are commonly used in cosmetic and personal care products. Such esters include, tocopheryl acetate, the tocopherol acetic acid ester; tocopheryl linoleate, the tocopherol linoleic acid ester; tocopheryl linoleate / oleate, a mixture of tocopherol linoleic and oleic acid esters; tocopheryl nicotinate, the tocopherol nicotinic acid ester; and tocopheryl succinate, the tocopherol succinic acid ester, potassium ascorbyl phosphate, a salt of both vitamin E (tocopherol) and vitamin C (ascorbic acid) can also be used in cosmetic products.
[0038] [0038] Other ingredients derived from tocopherol that can be found in cosmetic products include dioleyl tocopheryl methylsilanol, which is the dioleyl ether of tocopheryl acetate monoether with methylsilanotriol, and tocofersolane, which is also called polyethylene glycol 1000 detocoferyl succinate. of succinic acid and an average of 22 groups of ethylene oxide to tocopheryl makes tocofersolane a water-soluble form of tocopherol.
[0039] [0039] Mannitol is also an exemplifying stabilizer due to its low hygroscopic capacity, excellent chemical and physical drug compatibility, better sweetness and relatively slower dissolution kinetics. It also has relatively low water solubility and good dispersibility and is commonly used to improve the stability of the formulation where other excipients have failed. Mannitol can be used in a wide variety of dosage forms, including, but not limited to, tablets, capsules, sachets, lozenges, liquids, emulsions, suspensions, ointments, pastes, lotions and intravenous solutions.
[0040] [0040] Mannitol also serves as a matrix-forming additive for lyophilization. when used in concentrations of up to 10% by weight by volume, mannitol forms an amorphous (non-crystalline) matrix that supports proteins and other biomolecules for lyophilization. It is generally inert and, once lyophilized, it rehydrates quickly. Its amorphous structure when frozen prevents it from breaking down proteins while providing channels for sublimation of water during processing.
[0041] [0041] Similar to mannitol, sucrose is also widely used in tablet form for oral distribution due to its sweetness and palatability. Sucrose, is a non-reducing disaccharide (glucose bound by its anomeric carbon to fructose) that is widely used as a lyoprotectant.
[0042] [0042] Trealose (α-D-glycopyranosyl α-D-glycopyranoside), a disaccharide known for its antioxidant properties, was known as a non-reducing saccharide consisting of glycoses. As described in Advances in Carbohydrate Chemistry, Vol. 18, pp.201-225 (1963), published by Academic Press, USA, and Applied and Environmental Microbiology, Vol. 56, pp.3,213-3,215 (1990), trehalose it exists widely in microorganisms, mushrooms, insects, etc., although the content is relatively low. Trealose is a non-reducing saccharide so that it neither reacts with substances containing amino groups such as amino acids and proteins, induces the amino-carbonyl reaction and does not deteriorate the substances containing amino acids. In this way, trehalose can be used without fear of causing unsatisfactory browning and deterioration.
[0043] [0043] Trealose can also inhibit the inflammatory cascade, thereby suppressing cytokine production. (Minutoli, et al, SHOCK, Vol. 27, No. 1, pp. 91-96, 2007; and Chen Q, Haddad GG: Role of trehalose phosphate synthase and trehalose during hypoxia: from flies to mammals. J Exp Biol 207 : 3125-3129, 2004.) Trealose is a unique sugar capable of protecting biomolecules against environmental stress and can inhibit the inflammatory cascade, which in turn causes oxidative damage and the production of cytokines. It has also been shown that trehalose preserves cell viability during exposure to a range of environmental stresses, such as heat shock, dehydration and hypoxia.
[0044] [0044] Trealose is also a common food additive because it is a strong antioxidant and sweetener, and is commonly used as a stabilizing agent in pharmaceutical preparations. Trealose, like sucrose, is a non-reducing disaccharide (two glucose molecules linked by anomeric carbon) that can act as an effective lipoprotector for the lyophilization of proteins and other biomolecules. During the lyophilization process, proteins can denature as water is removed, unless a replacement molecule is available to support the protein's structure. Trehalose fills the void left by the water outlet and prevents this denaturation. When used in concentrations as low as 2%, it can effectively protect proteins and other biomolecules.
[0045] [0045] Useful forms of trehalose can include dihydrated trehalose (TD) which is crystalline, amorphous trehalose (AT) which is a glassy form, and anhydrous forms of trehalose, anhydrous amorphous trehalose (AAT) and anhydrous crystalline trehalose (ACT). Anhydrous powdered treats may contain AAT and / or ACT. The term "trehalose", for use in the present invention, refers to any physical form of trehalose including anhydrous, partially hydrated, completely hydrated and mixtures and solutions thereof. The manufacture and use of anhydrous TD trehalose can be found in international publication no: PCT / GB97 / 00367, the description of which is incorporated in this specification by reference.
[0046] [0046] The addition of trehalose to the formulation or composition can help stabilize high molecular weight HA, as well as inhibit harmful inflammatory cascades. Trehalose can be present in liquid, solid, lyophilized or crystalline forms. When present in liquid form, trehalose can be in a buffered solution. Solvents that can be used to solubilize trehalose may include, but are not limited to, water, saline or other salt solutions, buffer solutions such as phosphate buffered saline, histidine, lactate, succinate, glycine and glutamate, dextrose, glycerol , as well as combinations thereof. In particular, trehalose may be present in the HA formulation as a solution.
[0047] [0047] At least one stabilizer may be present in the HA formulation. Solvents that can be used to solubilize the stabilizer may include, but are not limited to, water, saline or other salt solutions, buffer solutions such as phosphate buffered saline, histidine, lactate, succinate, glycine and glutamate, dextrose, glycerol , as well as combinations thereof. In particular, the stabilizer may be present in the HA formulation as a solution.
[0048] [0048] The concentration of at least one stabilizer present in the HA formulation can vary, however in an exemplary embodiment, at least one excipient is provided in a pharmaceutically effective amount. In an exemplary embodiment, the at least one stabilizer has a concentration of at least about 1 mg / ml, and more preferably at least about 5 mg / ml, and more preferably at least about 50 mg / ml , and more preferably at least about 100 mg / ml, and in some embodiments the concentration can be at least about 200 mg / ml. Adequate concentrations of at least one stabilizer can include about 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 11 mg / mg, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, 20 mg / ml, 21 mg / ml, 22 mg / ml, 23 mg / ml, 24 mg / ml, 25 mg / ml, 26 mg / ml, 27 mg / ml, 28 mg / ml, 29 mg / ml, 30 mg / ml, 31 mg / ml, 32 mg / ml, 33 mg / ml, 34 mg / ml, 35 mg / ml, 36 mg / ml, 37 mg / ml, 38 mg / ml, 39 mg / ml, 40 mg / ml, 41 mg / ml, 42 mg / ml, 43 mg / ml, 44 mg / ml, 45 mg / ml, 46 mg / ml, 47 mg / ml, 48 mg / ml, 49 mg / ml, 50 mg / ml, 51 mg / ml, 52 mg / ml, 53 mg / ml, 54 mg / ml, 55 mg / ml, 56 mg / ml, 57 mg / ml, 58 mg / ml, 59 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml, 150 mg / ml, 160 mg / ml, 160 mg / ml, 170 mg / ml, 180 mg / ml, 190 mg / ml, 200 mg / ml, 300 mg / ml, 400 mg / ml, 500 mg / ml, 600 mg / ml or more hi any range d erivable in that. Stabilizers can also be in a concentration in the range of about 0.1 to 60%, by weight; from about 0.1 to 50% by weight, from about 0.1 to 45% by weight; or from about 0.1 to 20% by weight. Other suitable concentrations of at least one excipient may include about 0.1%, 0.5%, 1%, 2.5%, 5%, 6%, 7%, 8%, 9% 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% or about 60% by weight. Additional Components
[0049] [0049] In an exemplary embodiment, at least one additional component can also be combined with the HA formulation or composition. The additional component can be a glycosaminoglycan precursor (GAG) or lyophilized GAG. The term "glycosaminoglycan", or "GAG", refers interchangeably to the family of sulfated mucopolysaccharides that typically include heparin, heparin sulfate, chondroitin, chondroitin sulfate, keratan sulfate, dermatan sulfate and their derivatives.
[0050] [0050] Glycosaminoglycans (GAGs) are long unbranched polysaccharides containing a disaccharide and repeat unit. The disaccharide units contain either of the two modified sugars, N-acetylgalactosamine (GalNAc) or N-acetylglycosamine (GlcNAc), and a uronic acid such as glucuronate or iduronate. GAGs are highly negatively charged molecules, with extended conformation that gives the mixture a high viscosity. GAGs are located primarily on the cell surface or in the extracellular matrix (ECM). Along with the high viscosity of GAGs, there is low compressibility, which makes these molecules ideal for a lubricating fluid in the joints. GAGs can therefore help to slow down the inflammatory process. At the same time, their rigidity provides structural integrity to cells and provides passages between cells, allowing cell migration. In addition, a GAG like CS has been found in synovial fluid, and can play a role in joint health. Therefore, it may be advantageous to have a therapy that can release CS at the same time as HA. It is believed that CS can improve the effectiveness of HA. However, it has been observed that HA in the presence of CS can degrade over time. Therefore, in order to avoid this degradation, HA must be refrigerated until use. However, most clinics and hospitals do not have the ability to store these types of products in a refrigerator, so it could be beneficial to provide products that combine HA and GAGs that are stable and capable of being stored at room temperature.
[0051] [0051] GAGs can have several molecular weights, however in certain exemplifying modalities the molecular weight is in the range of about 5 to 1,000 KDa, more preferably in the range of about 6 to 500 KDa, more preferably in the range of about from 7 to 300 KDa, more preferably in the range of about 8 to 200 KDa, more preferably in the range of about 9 to 100 KDa and most preferably in the range of about 10 to 80 KDa. In other embodiments, the molecular weight of the GAG fragment is below about 5 KDa and, more preferably, below about 3 KDa.
[0052] [0052] The concentration of the GAGs present in the mixture can also vary, however in an exemplary embodiment the GAG is supplied in a pharmaceutically effective amount. In an exemplary embodiment, GAG has a concentration of at least about 0.1 mg / ml, and more preferably of at least about 2 mg / ml, and more preferably of at least about 5 mg / ml, and more preferably at least about 7 mg / ml, and more preferably at least about 20 mg / ml. In another embodiment, the GAG or GAG precursor may be present in the composition at a concentration in the range of about 0.1 mg / ml to about 20 mg / ml. Suitable concentrations of GAGs include about 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0 , 7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 11 mg / ml, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, 20 mg / ml, 21 mg / ml, 22 mg / ml, 23 mg / ml, 24 mg / ml, 25 mg / ml, 26 mg / ml, 27 mg / ml, 28 mg / ml, 29 mg / ml, 30 mg / ml, 31 mg / ml, 32 mg / ml, 33 mg / ml, 34 mg / ml, 35 mg / ml, 36 mg / ml, 37 mg / ml, 38 mg / ml, 39 mg / ml, 40 mg / ml, 41 mg / ml, 42 mg / ml, 43 mg / ml, 44 mg / ml, 45 mg / ml, 46 mg / ml, 47 mg / ml, 48 mg / ml, 49 mg / ml, 50 mg / ml, 51 mg / ml, 52 mg / ml, 53 mg / ml, 54 mg / ml, 55 mg / ml, 56 mg / ml, 57 mg / ml, 58 mg / ml, 59 mg / ml, 60 mg / ml or more or any derivable range thereof.
[0053] [0053] Chondroitin sulfate (CS), which is an essential component of cartilage, is composed of an alternating sequence of sulfated and / or not sulfated D-glucuronic acid (GlcA) and N-acetyl-D-galactosamine (GalNAc) sulfated compounds linked through alternating bonds β (1.3) and β (1.4). Each of these compounds has a polymeric structure consisting mainly of about 40 to 100 times the repetition of the disaccharide units. CS can be used in various molecular weights and concentrations, as discussed above in relation to the GAG component, however in an exemplary modality, CS has a molecular weight in the range of about 10,000 to 80,000 KDa and a concentration in the range of about 0 , 1 to 100 mg / ml. CS can be isolated from bovine or marine sources. A chondroitin chain can have more than 100 individual sugars, each of which can be sulfated in varying positions and amounts. Chondroitin sulphate 4, also carbon 4 in N-acetylgalactosamine sugar (GalNAc), is found in the cartilages of the nose and trachea of cattle and pigs. it is also found in the bones, flesh, blood, skin, umbilical cord and urine of these animals. Chondroitin sulphate 6, also carbon 6 from GalNAc sugar, was isolated from the skin, umbilical cord and heart valves of these animals. Chondroitin sulphate 6 has the same composition, but slightly different physical properties than chondroitin sulphate 4. Chondroitin sulphate is involved in collagen binding and is also directly involved in moisture retention. These two are properties that aid the healing process. One skilled in the art will appreciate that the terms "chondroitin sulfate", "CS", "chondroitin", "chondroitin sulfuric acid" and "chonsurid" are used interchangeably in the present invention and also encompass chemical or isomeric or cross-linked derivatives throughout this order.
[0054] [0054] Dermatan sulfate (DS), also called chondroitin sulfate B, is mainly composed of disulfated and / or trisulfated disaccharide units of L-iduronic acid and N-acetyl-D-galactosamine joined by β1,4 or 1 bonds , 3 but there is a case where some of the repeating units contain sulfated L-iduronic acid or D-glucuronic acid such as uronic acid, or contain unsulfated N-acetylgalactosamine or 4,6-disulfated N-acetylgalactosamine, instead of N -acetylgalactosamine-4-sulfate. DS is defined as a chondroitin sulfate by the presence of GalNAc. The presence of iduronic acid (IdoA) in DS distinguishes it from sulphates-A (4-O-sulphates of chondroitin) and -C (6-0-sulphates) and the alloy to heparin and HS, which also contains this residue. Dermatan sulfate is considered to be absorbed by the body when it is orally ingested. The molecular weight and concentration of DS can vary, as discussed above in relation to the GAG component, however in an exemplary embodiment the molecular weight is in the range of about 10 to 80 KDa and a concentration in the range of about 0.1 to 100 mg / ml. One skilled in the art will realize that, unlike HA, which is fermented by bacteria and therefore has a molecular weight that can be controlled, dermatan sulfate is isolated from animal tissue and may contain fragments. The molecular weight of dermatan sulfate, and any fragments contained therein, can therefore vary significantly. One skilled in the art will also realize that the terms "dermatan sulfate", "DS" and "dermatan" are used interchangeably in the present invention and also include sulfated derivatives of dermatan sulfate, the benzethonium salt of dermatan sulfate, the persulfated derivatives of the benzethonium salts of dermatan sulphate and also the sodium salt of dermatan sulphate.
[0055] [0055] Heparin and heparan sulfate (HS) are composed of a glucuronic acid (GlcA) linked to N-acetylglycosamine. They are compounds and linked α1-4 disaccharide repeat units containing a uronic acid and an amino sugar. Heparan sulfate proteoglycans are an integral part of the basement membrane. HS proteinoglycan is a large biomolecule with a molecular mass as high as 400 KDa, composed of a core protein covalently linked to heparan sulfate chains. The number of polysaccharide chains and the size of the nucleus protein can vary depending on the source. Heparan sulfate proteoglycan is a multifunctional molecule that binds to fibroblast growth factors, vascular endothelial growth factor (VEGF) and VEGF receptors through the sugar portion, acting as an anchoring molecule for matrilisin (MMP-7) and other matrix metalloproteinases that play important roles in cell proliferation and differentiation. Heparan sulfate proteoglians also promote cell binding by binding to a variety of molecules found in the extracellular matrix, including laminin, fibronectin, collagen type IV and FGF-basic. The molecular weight and concentration of HS can vary, as discussed above in relation to the GAG component, however in an exemplary embodiment the molecular weight is in the range of about 3 to 30 KDa (when isolated from tissues) and a concentration in the range of about 0.1 to 100 mg / ml.
[0056] [0056] Keratan sulfate, also keratosulfate (KS), is highly negatively charged and found mainly in agrecane, the most abundant proteoglycan in the extracellular matrix of hyaline, fibrous and elastic cartilage. KS is composed of the disaccharide repeat unit, -4GlcNAcβ1-3Galβ1-. Sulfation occurs at the carbon 6 (C6) position of either or both of the galactose (Gal) or GlcNAc. Specific KS types are composed of three regions: a binding region, at one end of which the KS chain is linked to the nucleus protein, the disaccharide repeat unit, and the chain covering region, which occurs at the opposite end of the KS chain in relation to the protein binding region. The molecular weight and concentration of KS can vary, as discussed above in relation to the GAG component, however in an exemplary embodiment the molecular weight is in the range of about 5 to 10 KDa (when isolated from tissues) and a concentration in the range of about from 1 to 100 mg / ml.
[0057] [0057] The concentration of GAG or GAG precursors present in the mixture can also vary, but in an exemplary embodiment the GAG or GAG precursor is supplied in a pharmaceutically effective amount. In an exemplary embodiment, the GAG or GAG precursor has a concentration of at least about 0.1 mg / ml, and more preferably at least about 2 mg / ml, and more preferably at least about 5 mg / ml, and more preferably at least about 7 mg / ml, and most preferably at least about 20 mg / ml. In another embodiment, the GAG or GAG precursor may be present in the composition at a concentration in the range of about 0.1 mg / ml to about 20 mg / ml. Suitable concentrations of the GAG or GAG precursors include about 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mg, 12 mg / mL, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, 20 mg / ml, 21 mg / ml, 22 mg / ml, 23 mg / ml, 24 mg / ml, 25 mg / ml, 26 mg / ml, 27 mg / ml, 28 mg / ml, 29 mg / ml, 30 mg / ml, 31 mg / ml, 32 mg / ml, 33 mg / ml, 34 mg / ml, 35 mg / ml, 36 mg / ml, 37 mg / ml, 38 mg / ml, 39 mg / ml, 40 mg / ml, 41 mg / ml, 42 mg / ml, 43 mg / ml, 44 mg / ml, 45 mg / ml, 46 mg / ml, 47 mg / ml, 48 mg / ml, 49 mg / ml, 50 mg / ml, 51 mg / ml, 52 mg / ml, 53 mg / ml, 54 mg / ml, 55 mg / ml, 56 mg / ml, 57 mg / ml, 58 mg / ml, 59 mg / ml, 60 mg / ml or more or any derivable range thereof.
[0058] [0058] One skilled in the art will realize that although lyophilized GAG or GAG precursors are particularly useful in exemplary embodiments, liquid GAG or GAG precursors can also be used in the composition. For example, CS can be obtained in powder form and mixed with a solvent, such as water, to form a solution. The solution can be combined with liquid HA to form a mixture having a reduced viscosity, compared to HA alone. Although it is effective in reducing the viscosity of HA, the resulting mixture will have a reduced concentration due to the presence of water. Thus, although non-lyophilized liquid GAGs can be used with the present invention, in an exemplary embodiment the GAG is lyophilized to allow the use of HA in a high concentration.
[0059] [0059] Glycosamine (C6H13NO5) ("GlcN") or its derivatives or other GAG precursors can also be included in the formulation of HA to enhance the synthesis of the key components of cartilage and synovial fluid by feeding both reactions necessary for production hyaluronan as well as for proteoglycans. GlcN is an amino sugar carrying four hydroxyl groups and an amine group, and it is a prominent precursor in the biochemical synthesis of glycosylated lipids and proteins. GlcN is a naturally occurring molecule that has nutrient and effector functions. For example, GlcN is compatible with and promotes the growth of stem cells and the differentiation of mesenchymal stem cells to form chondrocytes. GlcN may play a role in tissue development and repair, such as cartilage growth and development, in general. It is used as a nutritional supplement to combat OA symptoms, and has been shown to decrease the rate of cartilage destruction in clinical studies. In one embodiment, GlcN can be lyophilized together with CS. The glucosamine salt forms may have limited stability in the liquid phase. In addition, the GlcN HCl salt can lower the pH sufficiently to degrade HA once combined. For this reason, in order to retain the stability of the components, it is advantageous to have GlcN in lyophilized form to be solubilized with the HA gel before injection. For use in the present invention, "glucosamine" includes glucosamine salts, such as glucosamine hydrochloride and glucosamine sulfate, as well as non-salt forms like N-acetylglycosamine.
[0060] [0060] The GlcN concentration may vary. A suitable local concentration can be at least about 10 mg / ml, about 9 mg / ml, about 8 mg / ml, about 7 mg / ml, about 6 mg / ml, 5 mg / ml, 4 , 5 mg / ml, 4 mg / ml, 3.5 mg / ml, 3 mg / ml, about 2.9 mg / ml, about 2.8 mg / ml, about 2.7 mg / ml, about 2.6 mg / ml, about 2.5 mg / ml, about 2.4 mg / ml, about 2.3 mg / ml, about 2.2 mg / ml, about 2.1 mg / mL, about 2.0 mg / mL, about 1.9 mg / mL, about 1.8 mg / mL, about 1.7 mg / mL, about 1.6 mg / mL, about 1.5 mg / ml, about 1.4 mg / ml, about 1.3 mg / ml, about 1.2 mg / ml, about 1.1 mg / ml, about 1.0 mg / ml, about 0.9 mg / ml, about 0.8 mg / ml, about 0.7 mg / ml, about 0.6 mg / ml, about 0.5 mg / ml or so against. One skilled in the art can determine an appropriate local concentration of methods that practice GlcN known in the pharmaceutical art, and that the determination will govern the nature and composition of the GlcN composition of interest to obtain the desired GlcN concentration.
[0061] [0061] Buffering agents can also be added to the HA formulation to control the pH. Examples of buffering agents can be any one or more of the following agents, and are not limited to acetic acid, ammonium carbonate, ammonium phosphate, boric acid, citric acid, glycine, lactic acid, phosphoric acid, potassium citrate, metaphosphate potassium, monobasic potassium phosphate, sodium acetate, sodium citrate, sodium lactate solution, dibasic sodium phosphate, monobasic sodium phosphate, TRIS and sodium carbonate.
[0062] [0062] Additionally, isotonic agents can be added to control the ionic concentration and / or the osmotic pressure of the HA formulation. Examples of isotonic agents can be any one or more of the following agents, and are not limited to dextrose, sucrose, trehalose, glycerin, mannitol, potassium chloride and sodium chloride.
[0063] [0063] One skilled in the art will realize that the compositions and methods of the present invention may include several other components of joint treatment, including, for example, amino acids, proteins, saccharides, disaccharides, polysaccharides, lipids, nucleic acids, buffers, surfactants and mixtures thereof. Other useful components may include steroids, anti-inflammatory agents, non-steroidal anti-inflammatory agents, analgesic cells, antibiotics, microbicidal agents, anti-inflammatory agents, growth factors, growth factor fragments, small molecule wound healing stimulants, hormones, cytokines, peptides, antibodies, enzymes, isolated cells, platelets, immunosuppressants, nucleic acids, cell types, viruses, viral particles, essential nutrients or vitamins, and combinations thereof. Lyophilization
[0064] [0064] Any one or more of the components present in the compositions and methods of the present invention can be lyophilized using various techniques known in the art. Lyophilization is a dehydration process that is typically used to preserve a perishable material, and it works by freezing the material and then reducing the surrounding pressure and adding enough heat to allow the frozen water in the sublime material directly from the phase solid for the gas phase. Standard lyophilization techniques known in the art can be used to lyophilize any one or more of the components. In an exemplary embodiment, HA is lyophilized.
[0065] [0065] Before lyophilization, several solvents can be used to form an aqueous mixture containing the component (s) to be lyophilized. In an exemplary embodiment, the aqueous mixture is prepared by combining water with one or more of the components. In an exemplary embodiment, the composition is sterilized by filtration, as with a 0.2 pm filter, before lyophilization.
[0066] [0066] In one embodiment, the component (s) can be lyophilized (s) using the following cycle: Freezing: from room temperature to 5 ° C in 15 minutes Keep at 5 ° C for 100 minutes Lower the temperature to -45 ° C in 50 minutes Keep at -45 ° C for 180 minutes Primary Drying: adjust the pressure to 6.7 Pa (50 mTorr) Raise the temperature to -15 ° C in 175 minutes Keep at -15 ° C for 2300 minutes Secondary Drying: adjust the pressure to 10.0 Pa (75 mTorr) Raise the temperature to 25 ° C in 200 minutes Maintain the temperature for 900 minutes End of cycle: Refill with nitrogen to ~ 9.7 Pa (~ 730 Torr) Cap and curl
[0067] [0067] Variations for temperatures, times and adjustments can be made according to the practices used by one skilled in the art. Variations may include, but are not limited to, the cycling temperatures for the cycle and freezing, drying temperatures and final cycles. Variations can also include differences in waiting times for the freeze, dry and cap / crimp cycles. Variations may also include differences in pressures established for drying and capping / curling cycles. In addition, the number of drying cycles can be increased or decreased depending on the machine used or the component (s) to be lyophilized (s).
[0068] [0068] The addition of a buffering agent can provide improved protein solubility and stability in lyophilized HA formulations. Biocompatible buffering agents known in the art can be used, such as glycine; sodium, potassium or calcium acetate salts; sodium, potassium or calcium citrate salts; sodium, potassium or calcium lactate salts; sodium or potassium phosphate salts, including monobasic phosphate, dibasic phosphate, tribasic phosphate and mixtures thereof. Buffering agents may additionally have sugar added to the composition to function as a bulking agent. The pH preferably can be controlled to be between pH5 and pH10, and more preferably between pH 6 to 8. Formulations
[0069] [0069] In one embodiment, HA and at least one additional component, such as an excipient, can be configured to be combined and administered intrarticularly as part of a surgical procedure involving an articulated joint, just before, during or immediately after the surgical procedure. . Alternatively, HA and at least one additional component could be previously combined and present as a combination formulation at the time of the surgical procedure. The other components, when combined, can form a resulting composition or mixture having each component present in the composition in varying amounts. The amount of each component in the composition may vary, but in an exemplary embodiment, the mixing ratio between HA and at least one additional component, such as an excipient, may have a ratio between the weight of HA and the additional component in the range of about from 1: 0.001 to about 1: 100, and more preferably in a ratio in the range of about 1: 0.01 to about 1:10. Alternatively, the formulation can include about 1% to about 75% or more, by weight, of each of the individual components, such as HA and at least one additional component (for example, an excipient), in the total composition, alternatively about 2.5%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% or more, by weight, of HA and at least one additional component, as an excipient, in the formulation total. In an exemplary embodiment, the amount of HA present in the formulation presented can be about 1 to 20%, by weight, of the total formulation, and the amount of at least one additional component, such as an excipient, can be present in the formulation presented about 0.01 to 20% by weight of the total formulation.
[0070] [0070] Solvents that can be used to solubilize one or more of the components include, for example, water, acidic solvents, hydrochloric acid, acetic acid, benzoic acid, phosphate buffered saline, dextrose, glycerol, ethanol and the like, as well as their combinations. Solvents that can be used to solubilize HA and / or the additional component may include, but are not limited to, water, saline or other saline solutions, buffer solutions, such as phosphate buffered saline, histidine, lactate, succinate, glycine and glutamate, dextrose, glycerol, hydrochloric acid, acetic acid, benzoic acid, acidic solvent and other suitable solvents, as well as their combinations. The formulation can also include other components, such as dispersion medium, coatings, bactericidal and fungicidal agents, isotonic and delayed absorption agents and the like that are physiologically compatible. Isotonic agents include, for example, sugars, polyalcohols such as mannitol, sorbitol or sodium chloride in the composition. A formulation may also include minor amounts of adjuvant substances such as wetting or emulsifying agents, preservatives or buffers, which improve the life or effectiveness of the formulation.
[0071] [0071] The formulations can be incorporated into pharmaceutical compositions suitable for administration to an individual. Typically, the pharmaceutical composition comprises HA and another component, such as at least one excipient, and a pharmaceutically acceptable carrier. For use in the present invention, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, bactericidal and fungicidal agents, absorption retarding and isotonic agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, hydrochloric acid, acetic acid, benzoic acid, acidic solvent, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like, and combinations thereof. In many cases, it may be useful to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol or sodium chloride in the composition. Pharmaceutically acceptable vehicles may also comprise minor amounts of adjuvant substances such as wetting or emulsifying agents, preservatives or buffers, which improve the useful life or effectiveness of the composition.
[0072] [0072] Compositions can be in a variety of forms. These include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (for example, injectable and infused solutions), dispersions or suspensions, tablets, pills and powders. An exemplary form may depend on the intended mode of administration and the therapeutic application. Typical compositions can be in the form of injectable or infused solutions, such as compositions similar to those used for in vivo injection. In one embodiment, an exemplary mode of administration is parenteral (for example, intrarticular, subcutaneous, intraperitoneal, intramuscular). In another embodiment, the composition can be administered by infusion or injection directly into the target area, such as a joint. In yet another embodiment, the composition can be administered by intramuscular or subcutaneous injection.
[0073] [0073] Sterile injectable solutions can be prepared by incorporating the active compound in a therapeutically effective or beneficial amount in a suitable solvent with one or a combination of the ingredients listed above, as necessary, followed by filtration sterilization. In general, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the other necessary ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the modalities may include Preparation methods for vacuum drying and lyophilization which produce a powder of the active ingredient plus any desired additional ingredient from a solution previously sterilized by filtration thereof. Release Systems
[0074] [0074] The methods and compositions include kits for the treatment of joint disorders, such as the joints. The kits can comprise HA, and at least one stabilizer. Both and / or all components can be housed in a single chamber. The kit can also include HA with at least one stabilizer and an additional component. The additional component can be housed in a single chamber or separate from a syringe for injection with the mixture of HA and at least one stabilizer. HA and at least one stabilizer can be lyophilized. In an exemplary embodiment, HA is combined with at least one stabilizer in an individual chamber. In another embodiment, a kit is provided that has HA and at least one stabilizer housed in the same chamber and an additional component in a second chamber. In another exemplary embodiment, HA is combined with at least one stabilizer in the same chamber and a precursor to GAG or GAG is housed in a second chamber. The HA component can comprise up to about 10 ml of Orthovisc®, which contains about 150 mg of hyaluronan, 90 mg of sodium chloride and up to about 10.0 ml of water for injection and can be proportionally and appropriately reduced or increased for the specific indication. Examples can be for the knee, shoulder and hip, the injection volume for a single injection product can be in the range of about 3 ml to 10 ml and the hand can be in the range of about 500 pl and 1.5 ml . HA can have a molecular weight in the range of about 1.0 to 6 MDa. The at least one stabilizer present in the HA formulation or composition can have a concentration in the range of about 0.1 mg / ml to about 200 mg / ml. The at least one stabilizer can be lyophilized or, alternatively, it can be in a solution, as noted above, in combination with the HA component.
[0075] [0075] Compounds can be stored separately to increase service life. The individual compounds can be lyophilized or be in solid form in a syringe / cartridge with diluent or a second compound in a second syringe / cartridge. In one embodiment, one of the compounds is in lyophilized or solid form and the second compound is a solution capable of combining with the lyophilized / solid compound. An example can be at least one lyophilized or solid component that can be stored in a first chamber and a solubilized HA can be stored in a second chamber. In another embodiment, both compounds can be lyophilized or be in solid form and housed in a single chamber or in separate chambers of a syringe / cartridge. In another embodiment, compounds can be lyophilized directly in the syringe or cartridge.
[0076] [0076] Syringes and / or pre-filled double chamber cartridges can also be used with the components and compositions. Pre-filled double-chamber syringes allow sequential administration of two separate compositions with a single syringe push, thus replacing two syringes with one. The benefits of a single release capability include increasing the speed and ease of drug delivery; reduced risk of infection by reducing the number of connections; reducing the risk of drug and sequence administration errors, and faster distribution of compositions requiring combination prior to administration. The double-chamber syringe can accommodate lyophilized, powder or liquid formulations in the front chamber combined with diluents, saline or buffer in the rear chamber.
[0077] [0077] Pre-filled syringes can contain the exact releasable dose of the desired compounds and diluents. Pre-filled syringes can contain volumes of about 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml, 0.6 ml, 0.7 ml, 0.8 ml , 0.9 ml, 1.0 ml, 1.5 ml, 2 ml, 2.5 ml, 3 ml, 3.5 ml, 4 ml, 4.5 ml, 5 ml, 5.5 ml, 6 ml , 6.5 ml, 7 ml, 7.5 ml, 8 ml, 8.5 ml, 9 ml, 9.5 ml, 10 ml or more or any derivative thereof.
[0078] [0078] The syringe and / or double cartridge can be comprised of chambers side by side with separate syringe plungers that mix in a single chamber or linear chambers with a plunger. Syringes and / or double-chamber cartridges may also have a plug or connector in the middle to serve as a barrier between the two chambers. The plug or connector can be removed to allow mixing or combining of the compounds in the two chambers.
[0079] [0079] Figure 1 illustrates a modality of a mixing and delivery system that is in the form of a double-chamber syringe 10. As shown, the double-chamber syringe 10 generally includes a compartment having the proximal and distal chambers 14, 12 separated by a valve 16. A plunger 18 is slidably placed inside the proximal chamber 14 and is configured to inject fluid present inside the proximal chamber 14 into the distal chamber 12 in order to mix the components. In one embodiment, the first component, for example, liquid HA with at least one stabilizer, may be present in the proximal chamber 14 and an additional component, for example, one or more additional components, may be present in the distal chamber 12. Alternatively, the first component, for example, liquid HA, can be present in the proximal chamber 14 with at least one stabilizer, such as trehalose, and at least one additional component, for example, chondroitin sulfate, can be present in the distal chamber 12. The plunger 18 can advance through the proximal chamber 14 to inject the first component, for example, liquid HA with at least one stabilizer, into the distal chamber 12 containing the second component, for example, one or more additional components. In another embodiment, the proximal chamber 14 may contain a solvent, such as water or saline, and the distal chamber 12 may contain all components in solid form. For example, distal chamber 12 may contain lyophilized or solid HA with at least one stabilizer. The plunger 18 can advance through the proximal chamber 14 to inject the solvent into the distal chamber 12, thus solubilizing the components in the distal chamber 12. Once all the components are combined in the distal chamber 12, the mixture can be released into the tissue by example by attaching a needle to the distal end of the double-chamber syringe.
[0080] [0080] Figure 2 illustrates another modality of a mixing and release system 20, which is sold commercially under the trade name MixJect®. In this embodiment, the system includes a fluid controller assembly 22 that is coupled between a syringe 24 and a vial 26. Syringe 24 defines a first chamber 24a (not marked in the figure) that can contain a liquid, such as liquid HA or a solvent , and the vial defines a second chamber 26a (not marked in the figure) which may contain a solid, such as one or more additional components. Positioning the plunger 28 through syringe 24 will inject the liquid through the control system and into the vial 26, where the solid will be solubilized by the liquid. Once the components are completely solubilized, vial 26 can be inverted and plunger 28 can be retracted to pull the mixture back into chamber 24a in syringe 24. Vial 26 can then be removed from the system, and the mixture it can be injected from the syringe through a needle 29 and into the tissue.
[0081] [0081] One skilled in the art will realize that any double chamber systems known in the art can be used, and that the chambers can be chambers side by side with separate syringe plungers mixing in a single chamber or linear chambers with a single plunger. Treatments
[0082] [0082] method and compositions can be administered, for in vivo applications, parenterally by injection or by gradual perfusion over time. Administration can be intrarticular, intravenous, intraperitoneal, intramuscular, subcutaneous, intracavity or transdermal. For in vitro studies, agents can be added or dissolved in an appropriate biologically acceptable buffer and added to a cell or tissue.
[0083] [0083] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous vehicles include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered medium. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, intravenous Ringer's lactate vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present, for example, antimicrobials, antioxidants, chelating agents, growth factors and inert gases and the like.
[0084] [0084] Frequently used "vehicles" or "adjuvants" include magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatin, starch, vitamins, cellulose and its derivatives, oils of animal origin and vegetable oil, polyethylene glycols and solvents, such as sterile water, alcohols, glycerol and polyhydric alcohols, and dimethyl sulfoxide. Intravenous vehicles include fluid and nutrient replenishers. Preservatives include microbicidal agents, antioxidants, chelating agents and inert gases. Other pharmaceutically acceptable vehicles include aqueous solutions, non-toxic excipients, including salts, preservatives, buffers and the like, as described, for example, in Remington's Pharmaceutical Sciences, 15th ed. Easton: Mack Publishing Co .: 1405-1412, 14611487, 1975 and The National Formulary XIV., 14th ed. Washington: American Pharmaceutical Association, 1975 the contents of which are hereby incorporated by reference. The pH and the exact concentration of the various components of the pharmaceutical composition are adjusted according to routine skills in the art. See Goodman and Gilman's The Pharmacological Basis for Therapeutics (7th ed.).
[0085] [0085] Examples of symptoms or diseases, for which the composition and methods presented here may be useful, include the treatment of joint disorders, such as arthritis caused by infections, injuries, allergies, metabolic disorders, etc., rheumatoids such as arthritis chronic rheumatoid and systemic lupus erythematosus; joint disorders accompanied by gout, arthropathy such as osteoarthritis, internal breakdown, hydrarthrosis, stiff neck, lumbago, etc. By varying the effects depending on the use of the composition or the types of diseases to be treated, the agent can exert desired prophylactic and relief effects, or even therapeutic effects on swelling, pain, inflammation and destruction of the joints without seriously affecting living bodies. The composition for the treatment of joint disorders can be used to prevent the onset of joint disorders, as well as to improve, relieve and cure symptoms and their onset.
[0086] [0086] Treatment methods may include direct injection of the compositions into the target area, such as a joint. Injections can be performed as often as daily, weekly, several times a week, bimonthly, several times a month, monthly or as often as necessary to provide symptom relief. For intrarticular, the use of about 1 to about 30 mg / mL of HA per joint, depending on the size of the joint and the severity of the condition, can be injected. The frequency of subsequent injections into a given joint are spaced until the time the symptoms recur in the joint. Illustratively, human dosage levels of the composition can be: knee, about 1 to about 30 mg / mL by injection into the joint; shoulder, about 1 to about 30 mg / mL of HA by injection into the joint; metacorporal or proximal intrafalange, about 1 mg / mL to about 30 mg / mL HA by injection into the joint; and elbow, about 1 to about 30 mg / mL by injection into the joint. The total injection amount can be in the range of about 1 mg / ml to 200 mg / ml HA.
[0087] [0087] It will be understood, however, that the specific dosage level for any particular patient will depend on a variety of factors including the activity of the specific compound used, age, body weight, general health, sex, diet, length of stay administration, route of administration, excretion rate, drug combination and the severity of the particular disease being subjected to therapy. Pharmaceutical compositions can be prepared and administered in dose units. Under certain circumstances, however, higher or lower dose units may be appropriate. The administration of the dose unit can be carried out either by individual administration of the composition or the administration can be carried out in several smaller dose units and also by multiple administrations of doses subdivided at specific intervals.
[0088] [0088] In one embodiment, the medical condition is osteoarthritis (OA) and the composition is administered in a joint space, such as, for example, a knee, shoulder, temporomandibular joints and carpal-metacarpal joints, elbow, hip , wrist, ankle and lumbar zygapophyseal joints (facet) in the backbone. Viscosupplementation can be performed through a single injection or multiple intrarticular injections administered over a period of weeks in the knee or other distressed joints. For example, a human individual with OA in the knee may receive one, two, three, four or five injections of about 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9 , 10 mL or more per knee. For other joints, the volume administered can be adjusted based on the size of the joint.
[0089] [0089] It will be understood, however, that the specific dosage level for any particular patient will depend on a variety of factors including the activity of the specific compound used, age, body weight, general health, sex, diet, length of time administration, route of administration, excretion rate, drug combination and the severity of the particular disease being subjected to therapy. Experimental data Example 1: Stability of liquid CS and HA formulations with various stabilizers
[0090] [0090] HA at 2.5 mg / mL and chondroitin sulfate (CS) at 2.5 mg / mL were formulated with different stabilizers, as outlined in Table 1. The formulations were tested for the stability of HA under the conditions of storage at 5 ° C and 40 ° C / 75% relative humidity (Figure 3). After five months, the test samples were analyzed using a size exclusion chromatography (SEC) -high performance liquid chromatography method. Briefly, the test sample was diluted with a mobile phase (100 mM sodium phosphate buffer, pH 7) to 0.1 mg / ml HA. The diluted sample was then injected into an HPLC column (phenomenex linear column, Poly Sep-GFC-P, catalog number 00H-3147-K0) with a flow rate of 0.6 mL / min. The eluted HA peaks were monitored with a 207 nm wavelength and the retention time was compared with the HA molecular weight standards to determine the molecular weight of the test sample.
[0091] [0091] As shown in Table 1, the saline formulation was the least stable formulation at 40 ° C compared to the other samples at 40 ° C. Figure 3 shows the chromatograms of the saline formulation. The HA peak in the saline sample was shifted far to the right, indicating that HA has degraded into smaller molecules. In contrast, the trehalose formulation at 40 ° C had a much smaller displacement as shown in figure 4. Similar results were observed with the sucrose and mannitol formulations. These formulations should have a better shelf life at room temperature compared to the saline formulation. Table 1. Stability study of the HA and CS Formulation liquid at the five month time point
[0092] [0092] HA at 8 mg / mL and chondroitin sulfate at 8 mg / mL were also formulated with various stabilizers as outlined in Table 2. The formulated solutions were filled in 3 mL glass bottles at 0.5 mL / vial and lyophilized. The dry samples were placed at 40 ° C / 75% relative humidity for stability study. After 4 months, the test samples were reconstituted with WFI and analyzed by the SEC-HPLC method as mentioned in example 1. As shown in table 2, formulations containing saline were less stable compared to formulations containing glucose, mannitol, sucrose and trehalose. No significant changes in molecular weight were actually detected in these lyophilized formulations when stored at 40 ° C for four months. Table 2. Stability of the Lyophilized HA and CS Formulation at the four month time point
[0093] [0093] High concentrations of HA have also been tested. HA at 20 mg / mL and CS at 20 mg / mL formulated with various stabilizers. The formulations were tested for stability under storage conditions of 5 ° C and 40 ° C / 75% relative humidity as outlined in Table 3. After three months, the test samples were analyzed using the SEC-HPLC method . As shown in Table 3, the formulation containing saline only had less stability compared to formulations containing trehalose, sucrose and mannitol. Table 3. Stability of high concentration liquid HA and Cs formulations at the three-month time point
[0094] [0094] HA in different concentrations was formulated with various concentrations of trehalose, as outlined in Table 4. Some of the formulated solutions were divided into two groups. One was placed in stability chambers to study liquid stability and the other was lyophilized to assess solid stability. After three months of storage at 5 ° C and 40 ° C / 75% relative humidity, the test samples were analyzed using the SEC-HPLC method. As shown in Table 4, no significant difference in molecular weight was observed between samples with different concentrations of trehalose. However, without any excipients, HA formulated with water was only much less stable in liquid and lyophilized form. Table 4. Stability of the HA formulation with various concentrations of trehalose
[0095] [0095] In the middle rat meniscus (MMT) tear model of osteoarthritis, the median meniscus transection results in joint deterioration and reduced ability to support the weight that mimics human osteoarthritis. A tear of the unilateral median meniscus in 300 to 400 gram rats results in rapidly progressing degenerative cartilage changes characterized by loss of chondrocyte and proteoglycan, fibrillation, osteophyte formation and chondrocyte cloning. Such changes are typically seen to be substantial around the 21st day after meniscus surgery. The extent of joint deterioration, as determined primarily by the extent of the cartilage lesions formed, can be measured using a semi-quantitative histological scoring system.
[0096] [0096] To test the usefulness of trehalose as a vehicle for compounds injected intrarticularly, a trehalose solution was formulated as a 5% solution in 3 mM-HCl glycine buffer, having a pH of approximately 3. The solution was tested in mouse MMT model. Transection of the median collateral ligament just below the meniscus fixation was performed in 15 male Lewis rats. The meniscus was cut at its narrowest point (far from the ossicles) taking care not to damage the surface of the tibia.
[0097] [0097] Starting three days after MMT surgery, intrarticular injections of the trehalose solution were administered weekly for five weeks, followed by euthanasia in the sixth week.
[0098] [0098] Histological results showed that the trehalose solution provides an unexpected improvement in cartilage preservation. As shown in figure 5, trehalose provided a significant reduction in the width of significant cartilage lesions, as defined by the width of lesions in the tibial cartilage, in which the losses of chondrocytes and proteoglycans extend across 50% or more of the thickness of the original cartilage. Such measurements are performed using an ocular micrometer of stained histological cross sections of the median joint compartment.
[0099] [0099] Another measure of cartilage preservation is shown in Figure 6. Trehalose demonstrated a significant improvement in the lesion depth ratio, which is calculated by comparing the depth of the lesion with the original cartilage thickness, which is estimated by measuring from the distance from the original extrapolated tibial plateau to the footprint line. These measurements are performed using an ocular micrometer of stained histological cross sections of the median joint compartment.
[0100] [00100] Cartilage conservation also correlated with the extent of collagen degeneration. Figure 7 shows that trehalose significantly reduced the width of the cartilage lesions that exhibit collagen depletion, characterized as severe or severe, as measured using an ocular micrometer to assess the thickness of the cartilage cross section with reduced collagen placement. The extent of collagen degeneration is defined as outlined below: Serious injury (total or almost total loss of collagen to the mark,> 90% thick) Marked injury (extends through 61 to 90% of cartilage thickness) Moderate injury (extends across 31 to 60% of cartilage thickness) Moderate injury (extends across 11 to 30% of cartilage thickness) Minimal injury (very superficial, affecting only up to 10%)
[0101] [00101] Trealose also improved the classifications of cartilage degeneration, which is a measure of the cell viability of the remaining cartilage. Cartilage degeneration scores were determined histologically by estimating the area of non-viable cartilage matrix (significant loss of chondrocyte and proteoglycan, but collagen retention) and cartilage matrix completely absent, then by estimating the percentage of that area in compared to the area of the cross-sectional area of the original extrapolated cartilage. A score of 1 to 5 is assigned, with a score of 1 indicating cartilage degeneration of 10% or less, and a score of 5 indicating complete cartilage degeneration. In figure 8, zone 1 refers to one third of the median tibia plateau closest to the transfected meniscus. Zone 1 is the area that has the most severe cartilage damage in the MMT model, and was considered extremely challenging as a target for OA therapies. Zone 3 is the third part of the tibial plateau furthest from the surgical site, and sustained by the least amount of cartilage damage. Zone 2 was considered to be the area of damage to the intermediate cartilage and is represented by a reasonable target for OA therapies. Trehalose statistically improved the score of cartilage degeneration in all three zones. Example 4: Rabbit ACLT model of osteoarthritis
[0102] [00102] The trehalose formulation (a 5% trehalose solution in 3 mM-HCl glycine buffer with a pH of approximately 3) described above was also tested in the rabbit anterior cruciate ligament (ACLT) transection model of OA . ACLT surgery destabilized the rabbit's knee to a greater extent than the meniscus transection, resulting not only in substantial cartilage deterioration, but also in the formation of osteophytes. Studies investigating the development and regulation of osteophyte formation in the OA rabbit model detected plaque-like formations as early as 4 weeks after ACLT, and the growth of osteophytes in all knee compartments for 12 weeks after ACLT.
[0103] [00103] The rabbits were anesthetized. The patellar fat block was retracted, and the ACL was isolated. The anterior cruciate ligament was worsened with a surgical blade. The fibrous and subcutaneous capsule tissues were closed before the skin was sutured. Starting one week after surgery, intrarticular injections were administered weekly for seven weeks, followed by euthanasia for 8 weeks. Cartilage degradation was determined histologically using a modified Mankin scale. High scores indicated more extensive cartilage degradation, with a maximum score of 18.
[0104] [00104] As shown in figure 9, the trehalose formulation provided a significant improvement in the modified Mankin score compared to untreated animals. (n = 15), indicating decreased cartilage degeneration as a result of intrarticular trehalose injections. Terminology
[0105] [00105] A "therapeutically effective amount" or "effective amount" is that amount of an agent to achieve a pharmacological effect. The term "therapeutically effective amount" includes, for example, a prophylactically effective amount. An "effective amount" is an amount effective to achieve a desired pharmacological effect or therapeutic improvement without undue adverse side effects. For example, an effective amount refers to an amount that increases operability, quality of life, increases the patient's ability to increase mobility or increases the burden of bearing weight, or decreases pain, or increases growth in bone and cartilage of one or more joints, or reduces joint distortion, pain, swelling or stiffness. The effective amount of an agent will be selected by those skilled in the art depending on the patient and the level of the disease. It is understood that an "effective amount" or "a therapeutically effective amount" can vary from individual to individual, due to variation in the metabolism of therapeutic agents and / or prokinetic agents, age, weight, general condition of the individual, the health problem being treated, the severity of the health problem being treated and the judgment of the prescribing clinician.
[0106] [00106] "Treat" or "treatment" refers to any treatment of a disorder or disease associated with bone, soft tissue disorder cartilage (such as synovium), as preventing the occurrence of disorder or disease in an individual who may be predisposed to the disorder or disease, but who has not yet been diagnosed as having the disorder or disease; inhibition of the disorder or disease, for example, interruption of the development of the disorder or disease, relief of the disorder or disease, cause of regression of the disorder or disease, relief of a condition caused by the disease or disorder or interruption of the symptoms of the disease or disorder. Thus, for use in the present invention, the term "treat" is used in the same way as the term "avoid".
[0107] [00107] By "co-administered" is meant the simultaneous administration in the same formulation or in two different formulations that are combined in one formulation for administration. In one embodiment, HA and the other component are co-administered through application in the same formulation.
[0108] [00108] The term "individual", for use in the present invention, refers to an animal, in one embodiment, to a mammal and, in another embodiment, to a human being, who may benefit from the compositions and methods of the present invention . There is no limitation on the type of animal that could benefit from the present methods. An individual, regardless of whether he is a human or non-human animal, can be called an individual, subject, animal, host or recipient. The methods of the present invention have applications in human medicine, veterinary medicine, as well as in livestock in general, domestic or wild. In one embodiment, the candidate individual is a mammal such as a human being, a laboratory animal, such as a mouse, rat, rabbit, guinea pig, hamster, or bird species, such as a domestic bird and a veterinary animal, such as a dog, cat, horse, cow, sheep, etc.
[0109] [00109] skilled in the art will observe additional features and advantages of the invention based on the modalities described above. Accordingly, the invention should not be limited by what has been particularly shown and described, except as indicated by the appended claims. All publications and references cited herein are hereby expressly incorporated, by reference, in their entirety.

权利要求:
Claims (10)
[0001]
Composition for the treatment of a joint condition, characterized by the fact that it comprises: a formulation comprising high molecular weight hyaluronic acid (HA); at least one stabilizer comprising tocopherol or a tocopherol derivative, wherein the stabilizer increases the stability of the formulation, and at least one additional component, which is glycosaminoglycan (GAG) or a precursor of lyophilized GAG, wherein the additional component is present in the composition in an HA to additional component ratio in the range of 1: 0.005 to 1: 100.
[0002]
Composition according to claim 1, characterized by the fact that HA of high molecular weight has a molecular weight in the range of 1 MDa to 6 MDa.
[0003]
Composition according to claim 1 or 2, characterized by the fact that HA is present in a liquid concentration of at least 1 mg / mL.
[0004]
Composition according to any one of claims 1 to 3, characterized by the fact that the HA is lyophilized.
[0005]
Composition according to any one of claims 1 to 4, characterized in that the at least one stabilizer is present in the range of 0.1% to 50%, by weight of the composition.
[0006]
Composition according to any one of claims 1 to 5, characterized in that the formulation is stable at room temperature.
[0007]
Kit, characterized by the fact that it comprises: a high molecular weight hyaluronic acid (HA) composition, at least one stabilizer and at least one additional component, as defined in any one of claims 1 to 6; and a syringe comprising HA and stabilizer in a single chamber and a separate chamber comprising at least one additional component.
[0008]
Composition according to any one of claims 1 to 6, characterized by the fact that it is for use in the treatment of joints, which comprises administering said composition to an individual in need of it.
[0009]
Composition, according to claim 8, characterized by the fact that the administration step further comprises storing the composition at room temperature before administration.
[0010]
Composition according to claim 8 or 9, characterized in that the administration step further comprises combining HA and the stabilizer with at least one additional component before administration.

类似技术:

---

公开号 | 公开日 | 专利标题

---

BR102012016298B1|2020-09-24|STABILIZED POLYESACARIDE COMPOSITIONS FOR THE TREATMENT OF A JOINT AND KIT CONDITION

---

US11090328B2|2021-08-17|Compositions and methods for treating joints

---

US10532069B2|2020-01-14|Compositions and methods for treating joints

---

WO2008014685A1|2008-02-07|Formulation containing trehalose for intraarticular injection

---

WO2008014686A1|2008-02-07|Formulation containing low molecular heparin for intraarticular injection

---

US20150150983A1|2015-06-04|Intra-articular Formulations and Methods for Treatment of Osteoarthritis

同族专利:

---

公开号 | 公开日

---

EP2540284B1|2017-09-06|

---

EP2540284A3|2013-01-09|

---

EP2540284A2|2013-01-02|

---

EP3260116B1|2020-11-04|

---

CN102846653A|2013-01-02|

---

EP3260116A1|2017-12-27|

---

JP6113424B2|2017-04-12|

---

CN108125980A|2018-06-08|

---

US8623839B2|2014-01-07|

---

JP2013014586A|2013-01-24|

---

BR102012016298A2|2013-07-09|

---

US20140088038A1|2014-03-27|

---

US20130005681A1|2013-01-03|

---

CA2782122A1|2012-12-30|

---

AU2012203580B2|2016-03-31|

---

AU2012203580A1|2013-01-17|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

US3107605A|1960-09-23|1963-10-22|Colonial Sugar Refining Co|Extracting sucrose from sugar cane|

---

US3454560A|1966-03-01|1969-07-08|Seikagaku Kogyo Co Ltd|Process for the production of chondroitin polysulfate|

---

DE1792346C3|1968-08-22|1980-10-23|Rotta Research Laboratorium S.P.A., San Fruttuoso Di Monza, Mailand |Pharmaceutical preparation for the treatment of degenerative joint diseases|

---

US4141973B1|1975-10-17|1989-08-08|

---

US4971955A|1981-03-02|1990-11-20|Soll David B|Protection of human and animal cells during surgical trauma|

---

US4666897A|1983-12-29|1987-05-19|Research Foundation Of State University|Inhibition of mammalian collagenolytic enzymes by tetracyclines|

---

US5409904A|1984-11-13|1995-04-25|Alcon Laboratories, Inc.|Hyaluronic acid compositions and methods|

---

US4886786A|1985-08-01|1989-12-12|Lindstrom Richard L|Additive for irrigation solution or surgical solution|

---

US6051560A|1986-06-26|2000-04-18|Nestle S.A.|Chrondroitin sulfate/sodium hyaluronate composition|

---

JPH01113401A|1987-10-27|1989-05-02|Mitsubishi Rayon Co Ltd|Stabilization of aqueous hyaluronic acid solution|

---

US5266683A|1988-04-08|1993-11-30|Stryker Corporation|Osteogenic proteins|

---

US5011691A|1988-08-15|1991-04-30|Stryker Corporation|Osteogenic devices|

---

US5510418A|1988-11-21|1996-04-23|Collagen Corporation|Glycosaminoglycan-synthetic polymer conjugates|

---

US5013714A|1988-12-15|1991-05-07|Lindstrom Richard L|Viscoelastic solution|

---

US5380716A|1988-12-15|1995-01-10|Glycomed, Inc.|Sulfated polysaccharides as inhibitors of smooth muscle cell proliferation|

---

CA1340994C|1989-09-21|2000-05-16|Rudolf Edgar Dr. Falk|Treatment of conditions and disease|

---

US5246698A|1990-07-09|1993-09-21|Biomatrix, Inc.|Biocompatible viscoelastic gel slurries, their preparation and use|

---

US5143724A|1990-07-09|1992-09-01|Biomatrix, Inc.|Biocompatible viscoelastic gel slurries, their preparation and use|

---

US7025959B1|1992-02-12|2006-04-11|Biopharm Gesellschaft Zur Biotechnologischen Entwicklung Von Pharmaka Mbh|MP121, a growth/differentiation factor of the TGF-β family|

---

US5258371A|1992-05-29|1993-11-02|Kuraray Co., Ltd.|Method to reduce connective tissue destruction|

---

US5273056A|1992-06-12|1993-12-28|Alcon Laboratories, Inc.|Use of combinations of viscoelastics during surgery|

---

JP3390477B2|1993-01-25|2003-03-24|生化学工業株式会社|Pharmaceutical composition and method for producing the same|

---

US5364845C1|1993-03-31|2002-09-10|Nutramax Lab Inc|Glusosamine chondroitin and manganese composition for the protection and repair of connective tissue|

---

US6756358B2|1994-08-19|2004-06-29|Sumitomo Pharmaceuticals Co., Ltd.|Therapeutic agent for cartilaginous diseases|

---

IL116564D0|1995-01-04|1996-03-31|Mogen Int|Process for producing trehalose in plants|

---

US5792103A|1995-02-03|1998-08-11|Schwartz; Daniel M.|Viscosurgical method and apparatus|

---

IL114960D0|1995-03-20|1995-12-08|Medimop Medical Projects Ltd|Flow control device|

---

EA000582B1|1995-04-19|1999-12-29|Хехст Марион Руссел Лтд.|Protein, corresponding to a mature region of morphogenetic protein in human mp-52, pharmaceutical composition for treating of cartilage and bone diseases and method for preparing thereof|

---

US7026292B1|1995-12-12|2006-04-11|Stryker Corporation|Compositions and therapeutic methods using morphogenic proteins and stimulatory factors|

---

US7067144B2|1998-10-20|2006-06-27|Omeros Corporation|Compositions and methods for systemic inhibition of cartilage degradation|

---

JPH09176020A|1995-12-28|1997-07-08|Takada Seiyaku Kk|Stable solution containing sodium hyaluronate|

---

JPH09208476A|1996-01-26|1997-08-12|Takada Seiyaku Kk|Medicinal composition for abdominal administration|

---

US5789395A|1996-08-30|1998-08-04|The Research Foundation Of State University Of New York|Method of using tetracycline compounds for inhibition of endogenous nitric oxide production|

---

ZA9711580B|1996-12-25|1999-09-23|Hoechst Marion Roussel Ltd|Process for the production of purified dimeric bone morphogenetic factors.|

---

JP3748970B2|1997-01-31|2006-02-22|電気化学工業株式会社|Aqueous solution containing sodium hyaluronate|

---

US7223744B2|1997-02-10|2007-05-29|Inspire Pharmaceuticals, Inc.|Pharmaceutical formulation comprising dinucleoside polyphosphates and salts thereof|

---

US6818629B2|1997-02-10|2004-11-16|Inspire Pharmaceuticals, Inc.|Pharmaceutical formulation comprising P1-P4- tetraphosphate|

---

AU6267798A|1997-02-07|1998-08-26|Stryker Corporation|Matrix-free osteogenic devices, implants and methods of use thereof|

---

JPH1113401A|1997-06-26|1999-01-19|Mitsubishi Heavy Ind Ltd|Integral shroud moving blade|

---

KR100374666B1|1997-08-22|2003-03-04|덴끼 가가꾸 고교 가부시키가이샤|Hyaluronic acid gel, process for producing the same and medical material containing the same|

---

TW577758B|1997-10-27|2004-03-01|Ssp Co Ltd|Intra-articular preparation for the treatment of arthropathy|

---

US6541606B2|1997-12-31|2003-04-01|Altus Biologics Inc.|Stabilized protein crystals formulations containing them and methods of making them|

---

US6437018B1|1998-02-27|2002-08-20|Musculoskeletal Transplant Foundation|Malleable paste with high molecular weight buffered carrier for filling bone defects|

---

US7019192B2|1998-02-27|2006-03-28|Musculoskeletal Transplant Foundation|Composition for filling bone defects|

---

USRE38522E1|1998-02-27|2004-05-25|Musculoskeletal Transplant Foundation|Malleable paste for filling bone defects|

---

US6030635A|1998-02-27|2000-02-29|Musculoskeletal Transplant Foundation|Malleable paste for filling bone defects|

---

US7045141B2|1998-02-27|2006-05-16|Musculoskeletal Transplant Foundation|Allograft bone composition having a gelatin binder|

---

US6458375B1|1998-02-27|2002-10-01|Musculoskeletal Transplant Foundation|Malleable paste with allograft bone reinforcement for filling bone defects|

---

US6911212B2|1998-02-27|2005-06-28|Musculoskeletal Transplant Foundation|Malleable putty and flowable paste with allograft bone having residual calcium for filling bone defects|

---

US6326018B1|1998-02-27|2001-12-04|Musculoskeletal Transplant Foundation|Flexible sheet of demineralized bone|

---

US6410044B1|1998-03-19|2002-06-25|Surmodics, Inc.|Crosslinkable macromers|

---

WO1999051262A2|1998-04-03|1999-10-14|Chiron Corporation|Use of igfi for treating articular cartilage disorders|

---

JPH11302197A|1998-04-17|1999-11-02|Seikagaku Kogyo Co Ltd|Hyaluronic acid-stabilizing composition|

---

US6835377B2|1998-05-13|2004-12-28|Osiris Therapeutics, Inc.|Osteoarthritis cartilage regeneration|

---

EP2272535A1|1998-06-05|2011-01-12|Nutramax Laboratories, Inc.|The use of anabolic agents, anti-catabolic agents, antioxidant agents, and analgesics for protection, treatment and repair of connective tissues in humans and animals|

---

US6346519B1|1998-09-09|2002-02-12|Advanced Medical Instruments|Method and composition for treating arthritis|

---

US6656925B2|1998-09-09|2003-12-02|Advanced Medical Instruments|Composition and method of treating arthritis|

---

US7239908B1|1998-09-14|2007-07-03|The Board Of Trustees Of The Leland Stanford Junior University|Assessing the condition of a joint and devising treatment|

---

US6630457B1|1998-09-18|2003-10-07|Orthogene Llc|Functionalized derivatives of hyaluronic acid, formation of hydrogels in situ using same, and methods for making and using same|

---

WO2000021549A1|1998-10-14|2000-04-20|Orquest, Inc.|A method of inducing or enhancing chondrogenesis with extracellular matrix containing gdf-5|

---

IT1302534B1|1998-12-21|2000-09-05|Fidia Advanced Biopolymers Srl|INJECTABLE, BIOCOMPATIBLE AND BIODEGRADABLE COMPOSITIONS INCLUDING AT LEAST A DERIVATIVE OF HYALURONIC ACID, CHONDROGENIC CELLS, FOR|

---

US6727224B1|1999-02-01|2004-04-27|Genetics Institute, Llc.|Methods and compositions for healing and repair of articular cartilage|

---

JP2003089647A|1999-03-10|2003-03-28|Takada Seiyaku Kk|Articular disease therapeutic agent|

---

US6288043B1|1999-06-18|2001-09-11|Orquest, Inc.|Injectable hyaluronate-sulfated polysaccharide conjugates|

---

EP1074620A1|1999-08-06|2001-02-07|HyGene AG|Monomeric protein of the TGF-beta family|

---

US7651703B2|1999-10-15|2010-01-26|Genentech, Inc.|Injection vehicle for polymer-based formulations|

---

US7582311B1|1999-10-15|2009-09-01|Genentech, Inc.|Injection vehicle for polymer-based formulations|

---

DK1223990T3|1999-10-15|2004-11-29|Inst Genetics Llc|Formulations of hyaluronic acid for delivery of osteogenic proteins|

---

US6677321B1|1999-12-09|2004-01-13|Bruce Levin|Methods and compositions for treatment of inflammatory disease|

---

US20030086899A1|2000-03-14|2003-05-08|Jafari Masoud R.|Chondroitin sulfate containing viscoelastics for use in treating joints|

---

AT320262T|2000-05-12|2006-04-15|Bengt Krister Olson|COMPOSITIONS WITH COMBINED EXTRACTS FROM CORTIC TISSUE AND PLANTS|

---

US6924273B2|2000-10-03|2005-08-02|Scott W. Pierce|Chondroprotective/restorative compositions and methods of use thereof|

---

JP4754066B2|2000-12-22|2011-08-24|株式会社林原生物化学研究所|Anti-joint disorder|

---

US20040147466A1|2002-01-17|2004-07-29|Barman Shikha P.|Nucleic acid delivery formulations|

---

IL159624D0|2001-06-29|2004-06-01|Medgraft Microtech Inc|Biodegradable injectable implants and related methods of manufacture and use|

---

WO2003015755A1|2001-08-14|2003-02-27|Tel Aviv University Future Technology Development Lp|Lipidated glycosaminoglycan particles and their use in drug and gene delivery for diagnosis and therapy|

---

WO2003017826A2|2001-08-27|2003-03-06|Regeneration Technologies, Inc.|Processed soft tissue for topical or internal application|

---

HU0401283A2|2001-08-30|2004-12-28|Merckle Gmbh|Use of annellated pyrrole compounds in the treatment of articular cartilage or subchondral bone degeneration and pharmaceutical compositions containing them|

---

MXPA01011542A|2001-11-13|2003-05-22|Alcon Inc|Regeneration of articular cartilage damaged by osteoarthritis i and ii, by means of intra-articular application of sodium hyaluronate and chondroitin sulphate in a gel carrier.|

---

ES2383003T3|2001-11-21|2012-06-15|Carl Zeiss Meditec Sas|Composition to stabilize hyaluronic acid|

---

JP3779200B2|2001-11-28|2006-05-24|電気化学工業株式会社|Stabilized composition of aqueous solution of hyaluronic acid and / or salt thereof|

---

US20030181371A1|2001-12-28|2003-09-25|Angiotech Pharmaceuticals, Inc.|Compositions and methods of using collajolie|

---

EP1670500A2|2003-09-17|2006-06-21|Chiasma, Ltd.|Compositions capable of facilitating penetration across a biological barrier|

---

US7897174B2|2002-02-08|2011-03-01|University Of Erlangen-Nurnberg|Formulation comprising bioactive agents and method of using same|

---

US6855526B2|2002-05-15|2005-02-15|The United States Of America As Represented By The Secretary Of Agriculture|Method for making mannitol with Lactobacillus intermedius|

---

BR0310087A|2002-05-17|2005-08-16|Wyeth Corp|Hyaluronic acid injectable solid carriers for application of osteogenic proteins|

---

ES2353116T3|2002-07-16|2011-02-25|Bio Syntech Canada Inc.|COMPOSITIONS FOR CYCLOMPATIBLE, INJECTABLE, SELF-GRINDING QUITOSANE DISSOLUTIONS TO ENCAPSULATE AND ADMINISTER LIVING CELLS OR BIOLOGICALLY ACTIVE FACTORS.|

---

DE10246340A1|2002-10-04|2004-04-29|Wohlrab, David, Dr.|Combination preparation of hyaluronic acid and at least one local anesthetic and its use|

---

US7485629B2|2002-10-16|2009-02-03|Arthrodynamic Technologies, Animal Health Division, Inc.|Composition and method for treatment of joint damage|

---

EP1560588B1|2002-10-16|2011-09-21|ArthroDynamic Technologies, Animal Health Division, Inc.|Treatment for traumatic synovitis and damaged articular cartilage|

---

TWI235669B|2003-05-09|2005-07-11|Tty Biopharm Co Ltd|Concentrated emulsifier composition for preparing DB67 Silatecan injection solution|

---

US20050100538A1|2003-07-31|2005-05-12|Attawia Mohamed|Intradiscal injection of anti-oxidants|

---

US20090291112A1|2003-05-16|2009-11-26|Truncale Katherine G|Allograft osteochondral plug combined with cartilage particle mixture|

---

JP4694773B2|2003-06-06|2011-06-08|ロート製薬株式会社|Mucosal liquid composition|

---

US8361467B2|2003-07-30|2013-01-29|Depuy Spine, Inc.|Trans-capsular administration of high specificity cytokine inhibitors into orthopedic joints|

---

US8153591B2|2003-08-26|2012-04-10|Gel-Del Technologies, Inc.|Protein biomaterials and biocoacervates and methods of making and using thereof|

---

US20050058702A1|2003-09-17|2005-03-17|Ben-Sasson Shmuel A.|Compositions capable of facilitating penetration across a biological barrier|

---

US20050118230A1|2003-10-22|2005-06-02|Encelle, Inc.|Methods and compositions for regenerating connective tissue|

---

US7651682B2|2003-10-29|2010-01-26|Gentis, Inc.|Polymerizable emulsions for tissue engineering|

---

US20050250737A1|2003-11-12|2005-11-10|Allergan, Inc.|Therapeutic ophthalmic compositions containing retinal friendly excipients and related methods|

---

FR2866571B1|2004-02-20|2007-09-21|Philippe Zanchetta|USE OF A MIXTURE OF SPECIFIC POLYSACCHARIDES AS INDICATED BY THE EZBONE INVENTOR COMPRISING HYALURONIC ACID, CHONDROID SULFATE, DERMATANE SULFATE AND HEPARIN IN BONE HEALING.|

---

US20050232981A1|2004-04-15|2005-10-20|Ben-Sasson Shmuel A|Compositions capable of facilitating penetration across a biological barrier|

---

US7544671B2|2004-05-07|2009-06-09|S.K. Pharmaceuticals, Inc.|Stabilized hyaluronan preparations and related methods|

---

CA2570389A1|2004-06-17|2005-12-29|Osteologix A/S|Treatments comprising strontium for rheumatic and arthritic diseases and pain|

---

US20070286881A1|2005-07-14|2007-12-13|Brian Burkinshsw|Method, composition and kit for treating degenerated disc disease and discogenic pain|

---

CN101039683A|2004-08-13|2007-09-19|血管技术国际有限公司|Compositions and methods using hyaluronic acid and hyaluronidase inhibitors|

---

WO2006020994A2|2004-08-13|2006-02-23|Angiotech International Ag|Compositions and methods using hyaluronic acid and hyluronidase inhibitors|

---

BRPI0514482A|2004-08-18|2008-06-17|Biolog Health Solutions Pty Lt|herbal compositions for the prevention or treatment of urinary incontinence and overactive bladder|

---

US20060122150A1|2004-09-30|2006-06-08|Argentieri Dennis C|Pharmaceutical composition and method for treating a joint-capsule arthropathy|

---

DE102005034420A1|2004-12-23|2006-07-06|Ossacur Ag|Gel-like material for filling bone and / or cartilage defects|

---

KR20070104579A|2004-12-30|2007-10-26|젠자임 코포레이션|Regimens for intra-articular viscosupplementation|

---

ITPI20050071A1|2005-06-20|2006-12-21|Giuseppe Calvosa|COMPOUND COMPOSITION FOR FABRIC REPLACEMENT / REGENERATION|

---

WO2007004675A1|2005-07-06|2007-01-11|Seikagaku Corporation|Drug-containing photocrosslinked hyaluronic acid derivative gel|

---

RU2472809C2|2005-09-02|2013-01-20|Колбар Лайфсайенс Лтд.|Cross-linked polysaccharide and protein matrices and methods for production thereof|

---

CA2633957A1|2005-12-14|2007-06-21|Anika Therapeutics, Inc.|Treatment of arthritis and other musculoskeletal disorders with crosslinked hyaluronic acid|

---

GB2433518A|2005-12-21|2007-06-27|Danisco|Process for the recovery of sucrose and non-sucrose materials|

---

US8828977B2|2006-02-16|2014-09-09|Discogen, Llc|Method of treating a subject suffering from degenerative disc disease using a matrix metalloprotease inhibitor|

---

ITTO20060282A1|2006-04-14|2007-10-15|Univ Degli Studi Torino|MEDIUM OF CULTURE AND PHARMACEUTICAL COMPOSITION FOR THE REGENERATION OF THE RELATIVE PAPER FABRIC PROCEDURE RELATED TO USES AND PRODUCTS|

---

CN100571707C|2006-07-24|2009-12-23|凌沛学|The articular cavity inner injecting and administering preparations that contains trehalose|

---

KR20080024426A|2006-09-13|2008-03-18|유니메드제약주식회사|A stabilized hyaluronic acid derivative composition|

---

US7592009B2|2006-10-10|2009-09-22|Ecole Polytechnique Federale De Lausanne |Polypeptide ligands for targeting cartilage and methods of use thereof|

---

AU2007234612B2|2006-12-14|2013-06-27|Johnson & Johnson Regenerative Therapeutics, Llc|Protein stabilization formulations|

---

US8048857B2|2006-12-19|2011-11-01|Warsaw Orthopedic, Inc.|Flowable carrier compositions and methods of use|

---

CN101677957A|2007-02-05|2010-03-24|卡比兰生物外科公司|The polymer formulations that is used for delivery of bioactive agents|

---

DE102007030099A1|2007-06-28|2009-01-02|Henkel Ag & Co. Kgaa|Cosmetic composition containing a champagne extract|

---

US9011894B2|2007-06-29|2015-04-21|Carbylan Therapeutics, Inc.|Sterile hyaluronic acid polymer compositions and related methods|

---

FR2918276B1|2007-07-02|2010-01-22|Anteis Sa|"USE OF A NATURAL POLYSACCHARIDE GEL FOR THE PREPARATION OF AN INJECTION FORMULATION FOR THE TREATMENT OF JOINT DEGENERESCENCES"|

---

DE202007011252U1|2007-08-10|2007-12-20|Orthomol Pharmazeutische Vertriebs Gmbh|Amino sugar-containing product|

---

EP2033689A1|2007-08-22|2009-03-11|Italfarmacia S.r.l.|Injectable dermatological composition for treatment of the wrinkles|

---

US20090104148A1|2007-09-18|2009-04-23|Jay Gregory D|Treatment and prevention of joint disease|

---

US8986696B2|2007-12-21|2015-03-24|Depuy Mitek, Inc.|Trans-capsular administration of p38 map kinase inhibitors into orthopedic joints|

---

US20090162351A1|2007-12-21|2009-06-25|Depuy Spine, Inc.|Transdiscal administration of inhibitors of p38 MAP kinase|

---

WO2009100422A2|2008-02-08|2009-08-13|Zimmer, Inc.|Drug delivery system comprising microparticles and gelation system|

---

JP5757861B2|2008-04-24|2015-08-05|メドトロニック，インコーポレイテッド|Chitosan-containing protective composition|

---

JP5805542B2|2009-02-20|2015-11-04|オムリックス・バイオファーマシューティカルズ・リミテッドＯｍｒｉｘ Ｂｉｏｐｈａｒｍａｃｅｕｔｉｃａｌｓ Ｌｔｄ．|Device for administering at least two drugs|

---

KR101100803B1|2009-04-24|2011-12-29|서울과학기술대학교 산학협력단|Synthesis of lipoamide-grafted high molecular compound and method therefor|

---

US20110171310A1|2010-01-13|2011-07-14|Allergan Industrie, Sas|Hydrogel compositions comprising vasoconstricting and anti-hemorrhagic agents for dermatological use|

---

US8398611B2|2010-12-28|2013-03-19|Depuy Mitek, Inc.|Compositions and methods for treating joints|

---

US8455436B2|2010-12-28|2013-06-04|Depuy Mitek, Llc|Compositions and methods for treating joints|

---

US8524662B2|2010-12-28|2013-09-03|Depuy Mitek, Llc|Compositions and methods for treating joints|US8398611B2|2010-12-28|2013-03-19|Depuy Mitek, Inc.|Compositions and methods for treating joints|

---

US8524662B2|2010-12-28|2013-09-03|Depuy Mitek, Llc|Compositions and methods for treating joints|

---

US8455436B2|2010-12-28|2013-06-04|Depuy Mitek, Llc|Compositions and methods for treating joints|

---

TWI466675B|2011-09-16|2015-01-01|Univ China Medical|Pharmaceutical composition for inhibiting inflammation|

---

KR20140119363A|2013-03-29|2014-10-10|에스케이케미칼주식회사|Stable compositions of voriconazole|

---

EP3052083A4|2013-09-30|2017-05-03|Bioactive Regenerative Therapeutics, Inc.|Biomimetic hybrid gel compositions and methods of use|

---

KR101692884B1|2014-06-26|2017-01-04|마루이시세이야쿠가부시키가이샤|Rocuronium preparation with improved stability|

---

US10646574B2|2014-07-21|2020-05-12|Board Of Regents, The University Of Texas System|Formulations of intraarticular pharmaceutical agents and methods for preparing and using the same|

---

JP6427396B2|2014-11-19|2018-11-21|株式会社東芝|Magnetoresistance element and magnetic memory|

---

US9682099B2|2015-01-20|2017-06-20|DePuy Synthes Products, Inc.|Compositions and methods for treating joints|

---

EP3250251A1|2015-01-28|2017-12-06|Allergan, Inc.|Joint fat pad formulations, and methods of use thereof|

---

JP6847866B2|2016-01-29|2021-03-24|生化学工業株式会社|Stabilized aqueous composition containing chondroitin sulfate and hyaluronic acid|

---

EP3672653A4|2017-08-22|2021-01-13|Moebius Medical Ltd.|Liposomal formulation for joint lubrication|

---

EP3476384A1|2017-10-25|2019-05-01|F. Hoffmann-La Roche AG|Artificial vitreous humor for the investigation of drugs and drug formulations|

---

IT201800010415A1|2018-11-19|2020-05-19|Innate S R L|Injectable composition comprising hyaluronic acid and trehalose and use of said composition.|

法律状态:
2013-07-09| B03A| Publication of a patent application or of a certificate of addition of invention [chapter 3.1 patent gazette]|

---

2017-10-10| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|

---

2018-04-03| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

---

2018-07-24| B07E| Notification of approval relating to section 229 industrial property law [chapter 7.5 patent gazette]|Free format text: NOTIFICACAO DE ANUENCIA RELACIONADA COM O ART 229 DA LPI |

---

2019-07-23| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

---

2020-05-05| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

---

2020-09-24| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 29/06/2012, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

---

申请号 | 申请日 | 专利标题

---

US13/173,658|2011-06-30|

---

US13/173,658|US8623839B2|2011-06-30|2011-06-30|Compositions and methods for stabilized polysaccharide formulations|

---