HK1128593B - A composition suitable for ophthalmic administration - Google Patents

Description

Composition suitable for ophthalmic administration

Background

The present invention relates to the treatment of immunoinflammatory and ocular diseases.

Immunoinflammatory disorders are characterized by inappropriate activation of the body's immune defenses. The immune response does not target infectious invaders, but rather targets and destroys the body's own tissues or transplanted tissues. The tissues targeted by the immune system are altered by disease. For example, in multiple sclerosis, the immune response is directed to neural tissue, and in crohn's disease, to the digestive tract. Immunoinflammatory disorders affect millions of individuals and include diseases such as asthma, allergic intraocular inflammatory diseases, arthritis, atopic dermatitis, atopic eczema, diabetes, hemolytic anemia, inflammatory skin diseases, inflammatory bowel diseases or gastrointestinal diseases (e.g., crohn's disease and ulcerative colitis), multiple sclerosis, myasthenia gravis, pruritis/inflammation, psoriasis, rheumatoid arthritis, liver cirrhosis, and systemic lupus erythematosus.

Current treatment regimens for immunoinflammatory diseases typically rely on immunosuppressive agents. The effectiveness of these immunosuppressive agents can vary, and their use is often associated with adverse side effects. Accordingly, there is a need for improved therapeutic agents and methods for the treatment of immunoinflammatory disorders.

Musculoskeletal diseases such as arthritis are the most common cause of physical disability in the elderly. The three most common types of arthritis are Osteoarthritis (OA), Rheumatoid Arthritis (RA), and gout. Osteoarthritis is the most common joint disease, and radiological evidence of its presence is found in 50% of the population.

OA affects the hands, lower back, neck and weight bearing joints such as the knee, hip and foot joints. The annual incidence of OA in the hands is about 50 new cases/1,000 people under the age of 40, rising to 65 new cases/1,000 people at the age of 40-59, and 110 new cases/1,000 people at the age of 60 and above.

OA is characterized by a slowly progressing degenerative disease with a multi-factorial cause that may vary depending on joint location. OA occurs when cartilage (the tissue lining the bony ends of the joint) begins to break down and wear. In some cases, all of the cartilage may wear away, leaving the bones rubbed against each other. Arthroscopic studies of early disease have shown synovitis, with about half of those joints having cartilage damage, suggesting that the inflammatory response is localized in early OA patients. In addition, many studies have identified an association between C-reactive protein (CRP) and OA. CRP is an acute phase reaction protein whose production is stimulated by cytokines, particularly interleukin-6 (IL-6). The relationship between inflammatory processes and increases in plasma CRP and proinflammatory cytokines is known. CRP is also associated with the inflammatory activity of rheumatoid arthritis.

The symptoms of OA progress from rigidity and intermittent mild pain to severe joint pain and impaired biomechanical function. While there is no cure for most types of OA, various therapies can help patients maintain symptoms, improving their overall quality of life. Symptomatic treatment of OA has traditionally involved the administration of non-steroidal anti-inflammatory drugs (NSAIDs), local analgesic therapy, intra-articular injection of corticosteroids, and surgery.

Treatment of OA with NSAIDs, such as indomethacin, ketoprofen, ibuprofen, aspirin (ASA) and flurbiprofen, may alleviate pain by reducing local inflammation and reducing the level of proinflammatory agents. However, chronic NSAID use suffers from significant Gastrointestinal (GI) toxicity effects. A large multicenter, prospective, observational study involving 1,921 patients with rheumatoid arthritis taking NSAIDs reported that 81% of patients hospitalized for severe GI complications had previously had no GI problems. This makes it difficult for clinicians to identify patients at risk for GI side effects. In the united states, it has been properly estimated that there are 107,000 NSAID-associated GI complications hospitalized annually and 16,500 NSAID-associated deaths annually in RA or OA patients. This mortality rate is characterized by an almost very high number of deaths due to combined asthma, cervical cancer and malignant melanoma.

Steroids are known to be potent anti-inflammatory agents and have been used in the treatment of OA. However, the long-term administration of anti-inflammatory doses of steroids is also limited by known toxicity. For example, prolonged use of steroids has been associated with osteoporosis, hypertension, neurological complications, poor immune response and ocular disorders, limiting their utility in therapeutic situations. For example, a therapeutic agent that maintains the effective anti-inflammatory effect of a steroid or the therapeutic effect of another class of drugs would be very beneficial to patients with OA or other musculoskeletal disorders while limiting the associated toxicity.

Periodontal disease refers to the disease of the periodontal and dental supporting tissues including the gingiva, cementum, periodontal ligament, alveolar process bone and dental supporting bone. The most common forms of periodontal disease are periodontitis and gingivitis.

Periodontal disease involves inflammation, destruction, and degeneration of the periodontal tissue that surrounds and supports the teeth of mammals. These periodontal tissues include the crevice epithelium, the junctional epithelium, the outer marginal epithelium, the gingiva, the alveolar bone, the periodontal ligament and the cementum. The loss of supporting bone in periodontitis is a recent stage of the progressive disease and is a major cause of tooth loss in adults.

Periodontal disease is typically classified into gingivitis and periodontitis according to the progression of the disease. "gingivitis" refers to a disease in which inflammation is localized in the gingiva without damage to the bone and periodontal ligament, and the pockets are opposing pockets. "periodontitis" refers to a disease in which inflammation of the gingiva extends to the periodontal ligament and alveolar bone, and the pocket becomes a periodontal pocket, and the level of the attachment (the position of the attachment) is located on the apical side of the tooth root downward from the cement-enamel junction. The inflammation is prolonged and progresses to the deep part, and the periodontal pocket is deepened.

The relationship between inflammatory processes and elevations in plasma C-reactive protein (CRP) and proinflammatory cytokines is known and is observed in periodontitis. A correlation with the incidence of periodontitis has also been reported in atherosclerotic heart disease where elevated plasma CRP levels are a known risk factor. Thus, drugs that alter the level of inflammation of periodontitis would be expected to be effective against such inflammatory diseases. Indeed, treatment of periodontitis with NSAIDs, such as indomethacin, ketoprofen, ibuprofen, ASA, and flurbiprofen, may reduce local inflammation and reduce the level of proinflammatory agents. However, there has been no report on the use of corticosteroids for the treatment of periodontitis, perhaps because the known toxicity limits the long-term administration of anti-inflammatory doses of steroids. Drugs that can provide the anti-inflammatory effects of steroids without the associated toxicity are expected to be useful in the treatment of periodontal disease. Such a medicament would also be useful for lowering serum CRP levels and thus, may be useful in treating diseases and disorders associated with elevated serum CRP levels.

Summary of The Invention

The invention features compositions, methods, and kits for the treatment of immunoinflammatory and ocular diseases.

In one aspect, the invention features a composition that includes a drug pair selected from the drug pairs listed in table 1A and table 3. It is desirable that one or both drugs be present in amounts that together are sufficient to treat the following diseases in a patient in need thereof: immunoinflammatory disorders, ocular disorders or musculoskeletal disorders, or pain, tenderness, impaired motility, soft tissue swelling or bone swelling associated with these disorders. Exemplary drug pairs are: antihistamines and phosphodiesterase inhibitors; antihistamines and SSRIs; antihistamines and tricyclic compounds; anti-infective and anticoccidial compounds; corticosteroids and antihistamines; corticosteroids and phosphodiesterase inhibitors; corticosteroids and prostaglandins; NsIDI and alpha-2 adrenoreceptor agonists; NsIDI and antihistamines; NsIDI and NMDA antagonist/anti-dyskinesia agents; NsIDI and prostaglandins; NsIDI and sympathomimetic agents; prostaglandins and phosphodiesterase inhibitors; prostaglandins and tetra-substituted pyrimidopyrimidines; sympathomimetic and NMDA antagonist/anti-dyskinesia agents; sympathomimetic agents and prostaglandins; sympathomimetic agents and tetra-substituted pyrimidopyrimidines; sympathomimetic and tricyclic compounds; tetra-substituted pyrimidopyrimidines and phosphodiesterase inhibitors; tetra-substituted pyrimidopyrimidines and SSRIs; tetra-substituted pyrimidopyrimidine and tricyclic compounds; tricyclic compounds and calcium channel blockers.

One or both drugs in the pair may be present in the composition at a low dose or at a high dose. In certain embodiments, the compositions are formulated for local or systemic administration. In one embodiment, the composition is configured for ocular administration.

In another aspect, the invention features a method of: treating a patient diagnosed with an immunoinflammatory disorder by administering to the patient a drug pair selected from the group consisting of the drug pairs listed in table 1A or table 3. It is desirable to administer the first and second agents of the pair simultaneously or within 14 days of each other in amounts that together are sufficient to treat the patient. Optionally, one or more additional drugs (e.g., NSAIDs, corticosteroids, COX-2 inhibitors, biologicals, DMARDs, small molecule immunomodulators, xanthines, anticholinergic compounds, beta receptor agonists, bronchodilators, NsIDI, vitamin D analogs, psoralens, retinoids, or 5-amino salicylic acid) may also be administered to the patient.

In a related aspect, the invention features a method of: the immune response of the patient is modulated (e.g., by reducing secretion or production of pro-inflammatory cytokines, or by modulating adhesion, gene expression, chemokine secretion, presentation of MHC complexes, presentation of co-stimulatory signals, or cell surface expression of other modulators) by administering a pair of agents selected from the group consisting of the pairs of agents listed in table 1A to the patient simultaneously or within 14 days of each other in an amount sufficient to modulate the immune response of the patient.

In another aspect, the invention features a method of: treating a patient diagnosed with an ocular disease by administering to the patient a drug pair selected from the group consisting of the drug pairs listed in table 1A or table 3. It is desirable to administer the first and second agents of the pair simultaneously or within 14 days of each other in amounts that together are sufficient to treat the patient. Optionally, the patient may also be administered one or more additional drugs (e.g., anti-VEGF compounds, corticosteroids, NSAIDs, antiallergic agents, antihistamines, glaucoma-treating agents, antibiotics, antiviral agents, or antifungal agents) or receive photodynamic therapy.

In any of the foregoing methods, one or both of the agents in the pair is administered at a low dose or at a high dose. The drugs may be administered within 10 days of each other, within 5 days of each other, within 24 hours of each other, or simultaneously.

In a related aspect, the invention features a method of: treating the following diseases in a patient in need thereof by administering to the patient a pair of agents selected from the group consisting of the pairs listed in Table 1A together in an amount more effective than either agent alone in treating an immunoinflammatory disorder: immunoinflammatory disorders, ocular disorders, musculoskeletal disorders or pain associated therewith, periodontal disease or disorders associated with elevated serum CRP levels.

In yet another related aspect, the invention features a method of: treating the following disease in a patient in need thereof by administering to the patient a drug pair selected from the group consisting of the drug pairs listed in table 1A: immunoinflammatory disorders, ocular disorders, musculoskeletal disorders or pain associated therewith, periodontal disease or diseases or disorders associated with elevated serum CRP levels; wherein: in treating immunoinflammatory disorders, (i) both agents are administered simultaneously, and (ii) the respective amounts of both agents are administered to the patient more effectively than either agent alone.

In another aspect, the invention features a composition that includes: (i) a drug listed in table 1A or table 3; and (ii) a second compound selected from: anti-VEGF compounds, corticosteroids, NSAIDs, anti-allergic agents, antihistamines, glaucoma medication, antibiotics, antiviral agents, antifungal agents, COX-2 inhibitors, biologicals, DMARDs, small molecule immunomodulators, xanthines, anticholinergic compounds, beta receptor agonists, bronchodilators, NsIDI, vitamin D analogs, psoralens, retinoids or 5-amino salicylic acid.

In another aspect, the invention features a kit having (i) a composition including a drug pair selected from the group consisting of the drug pairs listed in table 1A or table 3; and (ii) instructions for administering the composition to a patient diagnosed with: immunoinflammatory disorders, ocular disorders, musculoskeletal disorders or pain associated therewith, periodontal disease or disorders associated with elevated serum CRP levels.

In a related aspect, the invention features a kit having (i) a first drug of a drug pair selected from the group consisting of the drug pairs listed in table 1A or table 3; (ii) a second drug of the same drug pair; and (iii) instructions for administering the two agents to a patient diagnosed with: immunoinflammatory disorders, ocular disorders, musculoskeletal disorders or pain associated therewith, periodontal disease or disorders associated with elevated serum CRP levels.

In another related aspect, the invention features a kit having (i) a first drug of a drug pair selected from the group consisting of the drug pairs listed in table 1A or table 3; and (ii) instructions for administering the drug and the second drug of the same drug pair to a patient diagnosed with: immunoinflammatory disorders, ocular disorders, musculoskeletal disorders or pain associated therewith, periodontal disease or disorders associated with elevated serum CRP levels.

In yet another related aspect, the invention features a kit having (i) a first agent listed in table 1A or table 3; (ii) a second agent selected from: anti-VEGF compounds, corticosteroids, NSAIDs, anti-allergic agents, antihistamines, glaucoma medication, antibiotics, antiviral agents, antifungal agents, COX-2 inhibitors, biologicals, DMARDs, small molecule immunomodulators, xanthines, anticholinergic compounds, beta receptor agonists, bronchodilators, NsIDI, vitamin D analogs, psoralens, retinoids or 5-amino salicylic acid; and (iii) instructions for administering said first agent and said second agent to a patient diagnosed with: immunoinflammatory disorders, ocular disorders, musculoskeletal disorders or pain associated therewith, periodontal disease or disorders associated with elevated serum CRP levels.

In another related aspect, the invention features a kit having (i) a first agent listed in table 1A or table 3; and (ii) instructions for administering the first agent and a second agent selected from the group consisting of: anti-VEGF compounds, corticosteroids, NSAIDs, anti-allergic agents, antihistamines, glaucoma medication, antibiotics, antiviral agents, antifungal agents, COX-2 inhibitors, biologicals, DMARDs, small molecule immunomodulators, xanthines, anticholinergic compounds, beta receptor agonists, bronchodilators, NsIDI, vitamin D analogs, psoralens, retinoids or 5-amino salicylic acid.

The invention also features methods, compositions, and kits for treating musculoskeletal disorders, or pain associated with musculoskeletal disorders, tenderness, impaired mobility, soft tissue distension, or bone distension in a patient in need thereof by administering a drug pair selected from the group consisting of the drug pairs listed in table 1A, optionally in combination with any one of a number of chaperone compounds, including corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs) (e.g., sodium naproxen, diclofenac potassium, aspirin, sulindac, diflunisal, piroxicam, indomethacin, ibuprofen, nabumetone, choline magnesium trisalicylate, sodium salicylate, salicylsalicylic acid, fenoprofen, flurbiprofen, ketoprofen, meclofenamate sodium, meloxicam, oxaprozin, sulindac, or tolmetin), COX-2 inhibitors (e.g., rofecoxib, celecoxib, valprozin, and/or a combination thereof), Valdecoxib or lumiracoxib), biologicals (e.g., adelimumab, etanercept, or infliximab), small molecule immunomodulators (e.g., VX 702, SCIO 469, doramapimod, RO30201195, SCIO 323, DPC333, pranalcasan, mycophenolate mofetil, or merimepodib), disease modifying antirheumatic drugs (DMARDs) (e.g., methotrexate or leflunomide), xanthines (e.g., theophylline), non-steroidal immunophilin-dependent immunosuppressants (NsIDI) (e.g., cyclosporine, tacrolimus, ascomycin, pimecrolimus, rapamycin, or everolimus), vitamin D analogs (e.g., calcipotriene or calcipotriol), psoralens (e.g., methoxsalen), retinoids (e.g., acitretin or tazoretene), 5-aminosalicylic acid (e.g., mesalamine, sulfasalazine, balsalazide disodium, or olsalazine sodium), hydroxychloroquine sulfate, penicillamine, or any of the analogs thereof as described herein.

Thus, in one aspect, the invention features a method of: by administering to a patient diagnosed with or at risk of developing: pain, tenderness, impaired mobility, soft tissue swelling, or bone swelling associated with musculoskeletal disorders such as osteoarthritis is treated by a drug pair selected from the drug pairs listed in Table 1A in an amount sufficient to treat the patient by administering the two drugs simultaneously or within 14 days, 10 days, 5 days, or even 24 hours of each other. It is expected that within 50 days of treatment, for example, the patient will have reduced pain, tenderness, impaired mobility, soft tissue swelling or bone swelling. Reduction in pain, tenderness, impairment in motility, soft tissue bulking or bone bulking can be measured using any method known in the art, such as a 10cm visual analog scale, a Likert scale, a Lequesne index or a WOMAC index. For example, the AUSCAN index using a 10cm visual analog scale can be used.

The invention also features the following method: a musculoskeletal disease, such as osteoarthritis, is treated by administering a drug pair selected from the group consisting of the drug pairs listed in table 1A to a patient diagnosed with or at risk of developing a musculoskeletal disease in an amount sufficient to treat the patient, either simultaneously or within 14 days, 10 days, 5 days, or even 24 hours of each other.

In any of the foregoing cases, the two drugs may be administered in the same or different pharmaceutical formulations. The compounds for use in the methods of the invention may be formulated for, e.g., topical or systemic administration, and may be formulated in high, medium or low doses.

In addition, a third agent, such as the following, may be administered to the patient: a corticosteroid, an NSAID, a COX-2 inhibitor, a biologic, a small molecule immunomodulator, a DMARD, a xanthine, an NsIDI, a vitamin D analog, a psoralen, a retinoid, 5-aminosalicylic acid, hydroxychloroquine sulfate, or penicillamine, such that the first agent, the second agent, and the third agent are administered simultaneously or within 14 days, 10 days, 5 days, or even 24 hours of each other in amounts sufficient to treat a patient.

In other embodiments, the agents of the pharmaceutical combinations of table 1A or table 3 are only two active ingredients (although excipients are typically also present).

In another aspect, the invention features a method of: a patient is treated for an ocular disease by administering a corticosteroid and a non-steroidal immunophilin-dependent immunosuppressant (NsIDI) to the patient. In this aspect of the invention, the corticosteroid and/or the NsIDI may be administered at low concentrations. It is desirable that the concentration of NsIDI does not cause ocular irritation, such as burning, and that the compositions of the present invention be administered in an amount sufficient to alleviate symptoms of ocular disease. It is also desirable that the concentration of corticosteroid does not cause toxicity of the steroid.

In another aspect, the invention features a method of: treating an ocular disease in a patient by administering to the patient an agent selected from the group consisting of: dipivefrin, anti-VEGF therapeutics, photodynamic therapy, NSAIDs, anti-allergic agents, antihistamines, glaucoma therapeutics, artificial tears, antibiotics, antiviral agents, and antifungal agents, in combination with corticosteroids and/or NsIDI. In this aspect of the invention, the corticosteroid and/or the NsIDI may be administered at low concentrations.

In another aspect, the invention features a composition (e.g., a solution, gel, cream, suspension, emulsion, or solid insert) that includes a corticosteroid and an NsIDI. In this aspect of the invention, the corticosteroid and/or the NsIDI may be administered at low concentrations.

In another aspect, the invention features a composition (e.g., a solution, gel, cream, suspension, emulsion, or solid insert) that includes a substance selected from the group consisting of: dipivefrin, anti-VEGF therapeutics, photodynamic therapy, NSAIDs, anti-allergic agents, antihistamines, glaucoma therapeutics, artificial tears, antibiotics, antiviral agents, and antifungal agents, in combination with corticosteroids and/or NsIDI. In this aspect of the invention, the corticosteroid and/or the NsIDI may be administered at low concentrations.

The invention also features a kit, comprising: (i) a corticosteroid; and (ii) instructions for administering a corticosteroid and an NsIDI to a patient having or at risk of having an ocular disease.

The invention also features a kit, comprising: (i) NsIDI; and (ii) instructions for administering a corticosteroid and an NsIDI to a patient having or at risk of having an ocular disease.

The invention also features a kit, comprising: (i) a composition comprising a corticosteroid and an NsIDI; and (ii) instructions for administering the composition to a patient having or at risk of having a metabolic disease.

The invention also features a kit, comprising: (i) a corticosteroid; (ii) NsIDI; and (iii) instructions for administering a corticosteroid and an NsIDI to a patient having or at risk of having an ocular disease.

Any of the foregoing kits may also include instructions for administering a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents.

Any of the foregoing kits may also include a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents. In these kits, the NsIDI and/or corticosteroid may optionally be formulated as a single composition having a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents.

The invention also features a kit, comprising: (i) a corticosteroid; and (ii) instructions for administering a corticosteroid and a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents.

The invention also features a kit, comprising: (i) NsIDI; and (ii) instructions for administering to a patient having or at risk of having an ocular disease an NsIDI and a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents.

The invention also features a kit, comprising: (i) a composition comprising a corticosteroid and a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutics, photodynamic therapy, NSAIDs, anti-allergic agents, antihistamines, glaucoma therapeutics, artificial tears, antibiotics, antiviral agents, and antifungal agents; and (ii) instructions for administering the composition to a patient having or at risk of having a metabolic disease.

The invention also features a kit, comprising: (i) a composition comprising NsIDI and a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutics, photodynamic therapy, NSAIDs, anti-allergic agents, antihistamines, glaucoma therapeutics, artificial tears, antibiotics, antiviral agents, and antifungal agents; and (ii) instructions for administering the composition to a patient having or at risk of having a metabolic disease.

The invention also features a kit, comprising: (i) a corticosteroid; (ii) a compound selected from: dipivefrin, anti-VEGF therapeutics, photodynamic therapy, NSAIDs, anti-allergic agents, antihistamines, glaucoma therapeutics, artificial tears, antibiotics, antiviral agents, and antifungal agents; (iii) instructions for administering a corticosteroid and a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents.

The invention also features a kit, comprising: (i) NsIDI; (ii) a compound selected from: dipivefrin, anti-VEGF therapeutics, photodynamic therapy, NSAIDs, anti-allergic agents, antihistamines, glaucoma therapeutics, artificial tears, antibiotics, antiviral agents, and antifungal agents; (iii) instructions for administering to a patient having or at risk of having an ocular disease an NsIDI and a compound selected from the group consisting of: dipivefrin, anti-VEGF therapeutic agents, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic agents, artificial tears, antibiotics, antiviral agents, and antifungal agents.

In any of the preceding aspects of the invention, the corticosteroid may be selected from: SEGRA (selective glucocorticoid receptor agonist), fluocinolone, fluorometholone, dexamethasone, hydrocortisone, loteprednol etabonate, medrysone, methylprednisolone, prednisolone, rimexolone, or triamcinolone.

In any of the preceding aspects of the invention, the NsIDI may be selected from cyclosporins A, ABT-281, ISAtx247, tacrolimus, ascomycin, pimecrolimus, rapamycin, or everolimus.

In any of the preceding aspects of the invention, the corticosteroid may be present at a concentration corresponding to that of prednisolone at a concentration of 0.01% to 0.1% (e.g., 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, or 0.01%) and the non-steroidal immunophilin-dependent immunosuppressant may be present at a concentration corresponding to that of cyclosporin a at a concentration of 0.001% to 0.049% (e.g., 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, or 0.001%).

In one embodiment, the corticosteroid is prednisolone, which is present at a concentration of 0.01% to 0.12% (e.g., 0.12%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is clocortolone valerate at a concentration of 0.01% to 0.1% (e.g., 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is hydrocortisone and the concentration of hydrocortisone is from 0.01% to 1.0% (e.g., 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is dexamethasone, and the concentration of dexamethasone is 0.01% to 0.1% (e.g., 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is fluorometholone at a concentration of 0.01% to 0.1% (e.g., 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is loteprednol etabonate at a concentration of 0.01% to 0.2% (e.g., 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is medroxypsone, at a concentration of 0.01% to 1.0% (e.g., 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In another embodiment, the corticosteroid is rimexolone at a concentration of 0.01% to 1.0% (e.g., 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

In any of the foregoing embodiments, the NsIDI may be cyclosporine a at a concentration of 0.001% to 0.049% (e.g., 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, and 0.001%).

In certain embodiments of the compositions, kits, and methods of the invention, only the pharmacologically active agent in the composition or kit or used in the method is recited. In this embodiment, pharmacologically inactive excipients may also be present in the composition or kit, or used in the practice of the method.

The invention features treatment of ocular diseases such as age-related macular degeneration, alkaline erosive keratoconjunctivitis, allergic conjunctivitis, allergic keratitis, anterior uveitis, behcet's disease, blepharitis, blood-aqueous barrier breakdown, choroiditis, chronic uveitis, conjunctivitis, contact lens-induced keratoconjunctivitis, corneal abrasion, corneal injury, corneal ulcer, crystalline retinopathy, cystoid macular edema, dacryocystitis, diabetic keratopathy, diabetic macular edema, diabetic retinopathy, dry eye disease, age-related dry macular degeneration, eosinophilic granuloma, episcleritis, exudative macular edema, Focus's dystrophy, giant cell arteritis, giant papillary conjunctivitis, glaucoma surgery failure, transplant rejection, herpes zoster, inflammation post cataract surgery, Iridocortical endothelial syndrome, iritis, keratoconjunctivitis sicca, keratoconjunctival inflammatory disease, keratoconus, lattice corneal dystrophy, map-dot-fingerprint corneal dystrophy, necrotizing keratitis, neovascular diseases involving the retina, uvea or cornea, such as neovascular glaucoma, corneal neovascularization, neovascularization after combined vitrectomy and phakotomy, neovascularization of the optic nerve and neovascularization due to penetrating or contusive eye injury, neuroparalytic keratitis, noninfectious uveitic ocular blepharitis, ocular lymphoma, ocular rosacea, ocular infection, ocular pemphigus, optic neuritis, panuveitis, optic papillary inflammation, planitis, persistent macular edema, phacoelerosis, posterior uveitis, Post-operative inflammation, proliferative diabetic retinopathy, proliferative sickle cell retinopathy, proliferative vitreoretinopathy, retinal artery occlusion, retinal detachment, retinal vein occlusion, retinitis pigmentosa, retinopathy of prematurity, rubeosis iritis, scleritis, Stewart-Johnson syndrome, sympathetic ophthalmia, temporal arteritis, thyroid-related eye disease, uveitis, vernal conjunctivitis, vitamin A deficiency-induced keratomalacia, vitritis, or age-related wet macular degeneration.

The compounds useful in the present invention include any pharmaceutically acceptable form of those compounds described herein, including isomers such as diastereomers and enantiomers, salts, esters, solvates, and polymorphs thereof, as well as racemic mixtures and pure isomers of the compounds described herein.

"corticosteroid" refers to any naturally occurring or synthetic compound characterized by the hydrogenated cyclopentane perhydro-phenanthrene ring system and having immunosuppressive and/or anti-inflammatory activity. Naturally occurring corticosteroids are generally produced by the adrenal cortex. Synthetic corticosteroids may be halogenated. Example corticosteroids are provided herein.

"non-steroidal immunophilin-dependent immunosuppressant" or "NsIDI" refers to any of the following non-steroidal drugs: decrease pro-inflammatory cytokine production or secretion, bind immunophilins or cause down-regulation of the pro-inflammatory response. NsIDI includes calcineurin inhibitors such as cyclosporins A, ABT-281, ISAtx247, tacrolimus, ascomycin, pimecrolimus, and other drugs (peptides, peptide fragments, chemically modified peptides or peptide mimetics) that inhibit the phosphatase activity of calcineurin. NsIDI also includes rapamycin (sirolimus) and everolimus, which bind to FK 506-binding protein, FKBP-12, blocking antigen-induced leukocyte proliferation and cytokine secretion.

"Small molecule immunomodulator" refers to the following non-steroidal, non-NsIDI compounds: reducing the production or secretion of pro-inflammatory cytokines, causing down-regulation of the proinflammatory response, or modulating the immune system in an immunophilin-independent manner. Exemplary small molecule immunomodulators are p38MAP kinase inhibitors such as VX 702(Vertex Pharmaceuticals), SCIO 469(Scios), doramapimod (Boehringer Ingelheim), RO30201195 (Roche) and SCIO 323(Scios), TACE inhibitors such as DPC333 (Bristol Myers Squibb), ICE inhibitors such as pranalcasan (Vertex Pharmaceuticals) and DH inhibitors such as mycophenolate (Roche) and merimepodib (IMP Pharmaceuticals).

"tricyclic compound" refers to a compound having one of formulas (I), (II), (III), (IV), or (V):

wherein each X is independently H, Cl, F, Br, I, CH₃、CF₃、OH、OCH₃、CH₂CH₃Or OCH₂CH₃(ii) a Y is CH₂、O、NH、S(O)_0-2、(CH₂)₂、(CH)₂、CH₂O、CH₂NH, CHN or CH₂S; z is C or S; a is a branched or unbranched saturated or monounsaturated hydrocarbon chain of between 3 and 6 carbons, including 3 and 6 carbons; each B is independently H, Cl, F, Br, I, CX₃、CH₂CH₃、 OCX₃Or OCX₂CX₃(ii) a D is CH₂O, NH or S (O)_0-2. In a preferred embodiment, each X is independently H, Cl or F; y is (CH)₂)₂Z is C; a is (CH)₂)₃(ii) a And each B is independently H, Cl or F. Other tricyclic compounds are described below. Tricyclic compounds include tricyclic antidepressants such as amoxapine, 8-hydroxyamoxapine, 7-hydroxyamoxapine, loxapine (e.g., loxapine succinate, loxapine hydrochloride), 8-hydroxyaloxapine, amitriptyline, clomipramine, doxepin, imipramine, trimipramine, desipramine, nortriptyline, and protriptyline, although it is contemplated that tricyclic compounds of the present invention need not have antidepressant activity.

"tetra-substituted pyrimidopyrimidines" refer to compounds having the formula (V):

wherein each Z and each Z' are independently N, O, C,Or

When Z or Z' is O orWhen Z or Z' is N, then p is 1,OrWhen Z or Z 'is C, then p is 2, and when Z or Z' is C, then p is 3. In the formula (V), each R₁Independently X, OH, N-alkyl (wherein the alkyl group has 1 to 20, more preferably 1 to 5 carbon atoms); branched or unbranched alkyl groups having 1 to 20, more preferably 1 to 5 carbon atoms; or heterocyclic, preferably as defined for formula (V). Or, when p > 1, two R of a common Z or Z' atom₁The radicals in combination may represent- (CY)₂)_k-, where k is an integer of 4 to 6 (including 4 and 6). Each X is independently Y, CY₃、C(CY₃)₃、CY₂CY₃、(CY₂)_1-5OY, Structure C_nY_2n-1Wherein n-3-7 (including 3 and 7). Each Y is independently H, F, Cl, Br or I. In one embodiment, each Z is the same moiety, each Z 'is the same moiety, and Z' are different moieties.

"prostaglandin" refers to a member of the lipid class of biochemical substances, belonging to a lipid subclass called eicosanoids, because of its structural similarity to C-20 polyunsaturated fatty acids, i.e., eicosenoic acids. The prostaglandins include prostaglandin E1, dinoprostone, limaprost, misoprostol, prostaglandin E2, prostaglandin a1, prostaglandin a2, prostaglandin B1, prostaglandin B2, prostaglandin D2, prostaglandin F1 α, prostaglandin F2 α, prostaglandin 11, prostaglandin-ici 74205, prostaglandin F2 β, 6-keto-prostaglandin F1 α, prostaglandin E1 ethyl ester, prostaglandin E1 methyl ester, prostaglandin F2 methyl ester, abaprost, ornoprost, 13, 14-dihydro prostaglandin F2 α, and prostaglandin J.

"butylbenzoic acid" refers to a compound having the structure:

or a pharmaceutically acceptable salt or prodrug thereof.

"Phenylbutadienoic acid analog" refers to a compound having the formula (VI):

or a pharmaceutically acceptable salt or prodrug thereof,

wherein R is¹Is composed ofOrWherein R is^1AAnd R^1BIs hydrogen, halogen, CF₃Optionally substituted C_1-6Alkyl, optionally substituted C_2-6Alkenyl, optionally substituted C_2-6Alkynyl, optionally substituted C_3-8Cycloalkyl, optionally substituted C_1-6Alkoxy or optionally substituted C_1-6A thioalkoxy group; r²And R³Each is independentIs immediately hydrogen and C_1-4Alkyl or CF₃；R⁴Is optionally substituted C_1-6Alkyl or optionally substituted C_3-8A cycloalkyl group.

"Low dose" or "low concentration" means at least 5% (e.g., at least 10%, 20%, 50%, 80%, 90%, or even 95%) less than the minimum standard recommended dose or minimum standard recommended concentration at which a particular compound is formulated for a given route of administration for the treatment of any human disease or condition. For example, a low dose of corticosteroid formulated for inhalation administration will be different than a low dose of corticosteroid formulated for oral administration.

By "high dose" is meant at least 5% (e.g., at least 10%, 20%, 50%, 100%, 200%, or even 300%) higher than the highest standard recommended dose of a particular compound for use in treating any human disease or disorder.

"Medium dose" refers to a dose between the low dose and the high dose.

"dose equivalent to the dose of prednisolone" refers to a dose of corticosteroid that, in combination with a given dose of a second drug, produces the same anti-inflammatory effect in a patient as the dose of prednisolone in combination with the given dose of the second drug.

"Selective 5-hydroxytryptamine reuptake inhibitor" or "SSRI" refers to any member of the following classes of compounds: (i) inhibits 5-hydroxytryptamine uptake by central nervous system neurons, (ii) has an inhibition constant (Ki) of 10nM or less, and (iii) has a selectivity for 5-hydroxytryptamine (i.e., a ratio of Ki (noradrenaline)/Ki (5-hydroxytryptamine)) of greater than 100 relative to selectivity for noradrenaline. Typically, when used as an antidepressant, the SSRI is administered at a dose greater than 10 mg/day. Exemplary SSRIs for use in the present invention are fluoxetine, fluvoxamine, paroxetine, sertraline, citalopram and venlafaxine.

"treatment" refers to the administration or formulation of a pharmaceutical composition for the treatment or prevention of an immunoinflammatory disorder.

"patient" refers to any animal (e.g., a human). Other animals that can be treated using the methods, compositions, and kits of the invention include horses, dogs, cats, pigs, goats, rabbits, hamsters, monkeys, guinea pigs, rats, mice, lizards, snakes, sheep, cattle, fish, and birds.

"steroid toxicity" refers to the significant detrimental increase in intraocular pressure resulting from administration of a steroid.

An "amount sufficient to" refers to the amount of the compound in the combination of the invention required to treat or prevent the immunoinflammatory disorder in a clinically relevant manner. The amount of active compound sufficient for use in the practice of the present invention to treat a condition caused by or attributable to an immunoinflammatory disorder varies with the mode of administration, the age, weight, and general health of the patient. Ultimately, the prescriber will determine the appropriate amount and dosage regimen.

By "more effective" is meant that the method, composition, or kit exhibits greater efficacy, less toxicity, is safer, more convenient, better tolerated, or is less expensive or provides a more satisfactory treatment than another method, composition, or kit of control. Efficacy may be determined by a skilled practitioner using any standard method appropriate for a given indication.

The term "immunoinflammatory disorder" includes a variety of conditions including autoimmune, proliferative and inflammatory skin disorders. Immunoinflammatory diseases result in the destruction of healthy tissue through inflammatory processes, dysregulation of the immune system, and unwanted proliferation of cells. Examples of immunoinflammatory disorders are acne vulgaris, acute respiratory distress syndrome, addison's syndrome, adrenocortical insufficiency, adrenogenital syndrome, allergic conjunctivitis, allergic rhinitis, allergic intraocular inflammatory disease, ANCA-associated small vessel vasculitis, angioedema, ankylosing spondylitis, aphthous stomatitis, arthritis, asthma, atherosclerosis, atopic dermatitis, autoimmune diseases, autoimmune hemolytic anemia, autoimmune hepatitis, behcet's disease, Bell's palsy, berylliosis, bronchial asthma, bullous dermatitis, bullous pemphigoid, myocarditis, abdominal disorders, cerebral ischemia, chronic obstructive pulmonary disease, liver cirrhosis, Cogan's syndrome, contact dermatitis, COPD, crohn's disease, cushing's syndrome, dermatomyositis, diabetes, discoid lupus erythematosus, and the like, Eosinophilic fasciitis, epicondylitis, erythema nodosum, exfoliative dermatitis, fibromyalgia, focal glomerulosclerosis, giant cell arteritis, gout, gouty arthritis, graft versus host disease, hand dermatitis, allergic purpura, herpes gestationis, hirsutism, hypersensitivity drug reaction, idiopathic kerato-scleritis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura, inflammatory bowel disease or gastrointestinal disease, inflammatory skin disease, juvenile rheumatoid arthritis, edema of larynx, lichen planus, Loeffler's syndrome, lupus nephritis, lupus erythematosus vulgaris, lymphoma tracheobronchitis, macular edema, multiple sclerosis, musculoskeletal and connective tissue diseases, myasthenia gravis, myositis, obstructive pulmonary disease, ocular inflammation, organ transplant rejection, osteoarthritis, pancreatitis, pemphigoid of pregnancy, herpes zoster, herpes simplex virus, and herpes simplex virus, Pemphigus vulgaris, polyarteritis nodosa, polymyalgia rheumatica, primary adrenocortical insufficiency, primary cirrhosis (primary bipolar cirrhosis), scrotal pruritus, pruritus/inflammation, psoriasis, psoriatic arthritis, Reiter's disease, recurrent polychondritis, rheumatic myocarditis, rheumatic fever, rheumatoid arthritis, rosacea due to sarcoidosis, rosacea due to scleroderma, rosacea due to Sweet's syndrome, rosacea due to systemic lupus erythematosus, rosacea due to urticaria, rosacea due to herpes zoster-related pain, sarcoidosis, scleroderma, segmental glomerulosclerosis, septic shock syndrome, seropathy, tendonitis or bursitis, Sjogren's syndrome, stills ' disease, stroke-induced brain cell necrosis, Sweet's disease, systemic dermatomyositis, chronic lymphocytic leukemia, chronic, Systemic lupus erythematosus, systemic sclerosis, Takayasu's arteritis, temporal arteritis, thyroiditis, toxic epidermal necrolysis, tuberculosis, type 1 diabetes, ulcerative colitis, uveitis, vasculitis, and Wegener's granulomatosis.

"non-skin inflammatory diseases" include rheumatoid arthritis, inflammatory bowel disease, asthma and chronic obstructive pulmonary disease.

"skin inflammatory disease" or "inflammatory skin disease" refers to an inflammatory disease selected from the group consisting of: psoriasis, guttate psoriasis, reversible (invert) psoriasis, pustular psoriasis, erythrodermic psoriasis, acute febrile neutrophilic dermatosis, eczema sicca, pompholyx, cystocematic palmoplantar eczema, acne vulgaris, atopic dermatitis, contact dermatitis, allergic contact dermatitis, dermatomyositis, exfoliative dermatitis, hand eczema, pompholyx, rosacea caused by sarcoidosis, rosacea caused by scleroderma, rosacea caused by Sweet's syndrome, rosacea caused by systemic lupus erythematosus, rosacea caused by urticaria, rosacea caused by herpes zoster-related pain, Sweet's disease, neutrophilic hidradenitis, aseptic pustulosis, drug eruptions, seborrheic dermatitis, pityriasis rosea, cutaneous lymphadenitis, pruritic papules and plaques of pregnancy, history-contract syndrome, and toxic necrolytic lysis, Tattoo response, Wells syndrome (eosinophilic cellulitis), reactive arthritis (Reiter's syndrome), gut-associated dermato logical arthritis syndrome, rheumatoid neutrophilic dermatosis, neutrophilic exocrine hidradenitis, dorsal-lateral hand neutrophilic dermatosis, plasmacytic localized balanitis, balanoposthitis, behcet's disease, centrifugal ringworm erythema, persistent cutaneous discoloring erythema, erythema multiforme, ringworm granuloma, hand dermatitis, lichen glossus, lichen planus, lichen sclerosus atrophicus, lichen simplex chronicus, lichen microtuberculus, nummular dermatitis, pyoderma gangrenosum, sarcoidosis, sub-cuticular pustular dermatosis, urticaria and transient acanthosis.

"proliferative skin disease" refers to a benign or malignant disease characterized by accelerated cell division of the epidermis or dermis. Examples of proliferative skin diseases are psoriasis, atopic dermatitis, non-specific dermatitis, primary irritant contact dermatitis, allergic contact dermatitis, actinic keratosis, basal cell carcinoma and squamous cell carcinoma of the skin, ichthyosis lamellar, epidermolytic hyperkeratosis, pre-malignant keratosis, acne and seborrheic dermatitis.

"musculoskeletal disease" refers to a disease associated with the immune system of muscle, ligament, bone, joint, cartilage, or other connective tissue. The most common musculoskeletal disorders occur in different forms of arthritis, such as osteoarthritis, rheumatoid arthritis, juvenile rheumatoid arthritis, and gout. Other musculoskeletal diseases include acquired hypertrophic bone syndrome, acromegaly, ankylosing spondylitis, Behcet's disease, bone disease, bursitis, osteomalacia, chronic fatigue syndrome, compartment syndrome, congenital hypothyroidism, congenital myopathy, odontoseisis, dermatomyositis, diffuse idiopathic hyperostosis, Dupuytren's spasm, hypereosinophilic myalgia syndrome, fasciitis, Felty's syndrome, fibromyalgia, hallux valgus, infectious arthritis, joint disease, dancing cosmetic syndrome, juvenile deformatitis, lupus, Lyme disease, Melas syndrome, metabolic bone disease, mitochondrial myopathy, mixed connective tissue disease, muscle disease, muscular atrophy, musculoskeletal deformity, musculoskeletal disease, myositis, ossification fasciitis, necrotizing fasciitis, neuroarthropathy, osteitis deformans, osteomyelitis, Osteochondrosis, osteomalacia, osteomyelitis, osteonecrosis, osteoporosis, Paget's disease, Pierre Robin syndrome, polymyalgia rheumatica, polymyositis, post-polio syndrome, pseudogout, psoriatic arthritis, reactive arthritis, Reiter's disease, relapsing polychondritis, renal osteodystrophy, rhabdomyolysis, rheumatism, rheumatic fever, scleroderma, Sever's disease (calcaneal neuritis),Syndrome, spondylopathy, spinal stenosis, stills ' disease, synovitis, temporomandibular joint disease, tendinopathy, inflammation of the elbow, tenosynovitis, tietz's syndrome, and Wegener's granulomatosis.

"ocular disease" refers to physiological abnormalities of the eye that may involve the retina, vitreous, lens, cornea, sclera, or other parts of the eye, or adversely affect physiological abnormalities of the eye such as inadequate tear production.

Ocular diseases that may be treated with the compositions, methods, and kits of the invention include age-related macular degeneration, alkaline-erosive keratoconjunctivitis, allergic conjunctivitis, allergic keratitis, anterior uveitis, behcet's disease, blepharitis, blood-aqueous barrier rupture, choroiditis, chronic uveitis, conjunctivitis, contact lens-induced keratoconjunctivitis, corneal abrasion, corneal injury, corneal ulcer, phakic retinopathy, macular cystoid edema, dacryocystitis, diabetic keratopathy, diabetic macular edema, diabetic retinopathy, dry eye disease, age-related dry macular degeneration, eosinophilic granuloma, episcleritis, exudative macular edema, fox's dystrophy, giant cell arteritis, giant papillary conjunctivitis, glaucoma surgery failure, transplant rejection, glaucoma, ocular inflammation, ocular, Herpes zoster, post-cataract surgery inflammation, iridocorneal endothelial syndrome, iritis, keratoconjunctivitis sicca, keratoconjunctival inflammatory disease, keratoconus, lattice corneal dystrophy, map-dot-fingerprint corneal dystrophy, necrotizing keratitis, neovascular diseases involving the retina, uvea or cornea such as neovascular glaucoma, corneal neovascularization, neovascularization after combined vitrectomy and phakotomy, neovascularization of the optic nerve and neovascularization due to penetrating or contusion eye injury, neuroparalytic keratitis, noninfectious uveitis inflammatory eye herpes, ocular lymphoma, ocular rosacea, ocular infection, ocular pemphigus, optic neuritis, panuveitis, optic papillary inflammation, pars plana, persistent macular edema, Phacoanaphylaxis, posterior uveitis, post-operative inflammation, proliferative diabetic retinopathy, proliferative sickle cell retinopathy, proliferative vitreoretinopathy, retinal artery occlusion, retinal detachment, retinal vein occlusion, retinitis pigmentosa, retinopathy of prematurity, rubeosis iritis, scleritis, Stewart-Johnson syndrome, sympathetic ophthalmia, temporal arteritis, thyroid-related eye disease, uveitis, vernal conjunctivitis, vitamin A deficiency-induced keratomalacia, vitritis, or age-related wet macular degeneration.

As recognized by those skilled in the art, a particular disease, disorder or condition may be characterized by both proliferative and inflammatory skin diseases. An example of such a disease is psoriasis.

It is also recognized that a particular disease may be either an immunoinflammatory disease or an ocular disease. One such example is Behcet's disease.

The term "periodontal disease" includes a variety of diseases including gingivitis and periodontitis, as well as diseases of the tissues surrounding and supporting the teeth including the gums, cementum, periodontal ligament, alveolar bone and dental supporting bone.

"disease or condition associated with elevated serum CRP levels" refers to any disease or condition that may have higher serum CRP levels than normal controls. Typically, serum CRP levels are considered elevated at > 3 mg/L. Such diseases and conditions associated with elevated serum CRP levels include cardiovascular disease (e.g., coronary artery disease, peripheral artery disease), hypertension, colon cancer, lymphoma, sarcoma, and pancreatitis.

"sustained release" or "controlled release" refers to the release of a therapeutically active ingredient from a formulation at a controlled rate, thereby maintaining therapeutically beneficial blood levels (but below toxic levels) of the ingredient over an extended period of time, e.g., about 12 to about 24 hours, thus providing a dosage form for, e.g., 12 hours or 24 hours.

In the general description of the compounds of the invention, the number of atoms of a particular type in a substituent is generally given in the range, e.g. alkyl having 1 to 4 carbon atoms or C_1-4An alkyl group. Reference to such ranges is intended to include a specific reference to the radical having an integer number of atoms in the specified rangeAnd (4) clustering. For example, alkyl of 1 to 4 carbon atoms includes C₁、C₂、C₃And C₄Each of which. C_1-12Heteroalkyl groups include, for example, 1 to 12 carbon atoms in addition to 1 or more heteroatoms. Other numbers of atoms and other types of atoms may be indicated in a similar manner.

As used herein, the term "alkyl" and the prefix "alk-" include straight-chain, branched-chain groups and cyclic groups, i.e., cycloalkyl groups. The cyclic group can be monocyclic or polycyclic, preferably having from 3 to 6 (including 3 and 6) ring carbon atoms. Exemplary cyclic groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

″C_1-4Alkyl "refers to a branched or unbranched hydrocarbon group having 1-4 carbon atoms. C_1-4Alkyl groups may be substituted or unsubstituted. Exemplary substituents include alkoxy, aryloxy, mercapto, alkylthio, arylthio, halide, hydroxy, fluoroalkyl, perfluoroalkyl, amino, aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl, carboxyalkyl and carboxy. C_1-4Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and cyclobutyl.

"halogen" refers to bromine, chlorine, iodine or fluorine.

"alkoxy" refers to a chemical substituent of the formula-OR, wherein R is selected from C_1-7Alkyl radical, C_2-7Alkenyl radical, C_2-7Alkynyl, C_2-6Heterocyclic group, C_6-12Aryl radical, C_7-14Alkylaryl group, C_3-10Alkylheterocyclic group or C_1-7A heteroalkyl group.

The term "pharmaceutically acceptable salts" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are known in the art. The salts may be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reaction of the free base functionality with a suitable organic acid. Representative acid addition salts include: acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate (heptanate), hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, isethionate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectate, persulfate, 3-phenylpropionate, Phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, tosylates, undecanoates, valerates, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts and the like, as well as non-toxic ammonium, quaternary ammonium and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine and the like.

The compounds useful in the present invention include any pharmaceutically acceptable form of those compounds described herein, including isomers such as diastereomers and enantiomers, salts, esters, amides, thioesters, solvates, and polymorphs thereof, as well as racemic mixtures and pure isomers of the compounds described herein.

Brief Description of Drawings

FIGS. 1A-1MM are graphs showing PMA/ionomycin-induced inhibition of TNF α secretion in cells treated with drug pairs selected from the drug pairs listed in Table 1A or Table 3.

FIGS. 2A-2LL are graphs showing the inhibition of IFN γ, IL-2, and TNF α in cells treated with a combination of NsIDI and a corticosteroid.

Other features and advantages of the present invention will be apparent from the following detailed description.

Detailed description of the invention

The invention features methods, compositions, and kits for the treatment of immunoinflammatory disorders. In one embodiment of the invention, the treatment of immunoinflammatory disorders is carried out by administering both agents simultaneously or within 14 days of each other to a patient in need of such treatment.

The invention also features methods, compositions, and kits for the treatment of ocular diseases. In one embodiment, a patient suffering from an ocular disease is treated by administering two drugs simultaneously.

Cyclosporin a, a non-steroidal immunophilin-dependent immunosuppressant (NsIDI), is approved for the treatment of several ocular diseases. Cyclosporin a causes eye irritation and other undesirable side effects when administered to patients at the lowest approved concentration. Lower concentrations of cyclosporin a do not cause these undesirable side effects, but are not sufficient to alleviate symptoms of ocular disease.

Both corticosteroids and NsIDI inhibit cytokine production in cell culture models of immune function (table 1B). We have found that: the combination of certain NsIDI with certain corticosteroids inhibits cytokine production in a synergistic manner.

Based on these data, we propose: when combined with corticosteroids, low concentrations of cyclosporin a are sufficient to alleviate symptoms of ocular diseases without causing undesirable side effects.

The present invention is based on our findings: certain combinations of drugs inhibit the production and/or secretion of cytokines in vitro. These combinations are listed in tables 1A and 1B.

TABLE 1A

Medicine 1	Medicine 2	Medicine 1	Medicine 2
				Prednisolone	Ibudilast	Ibudilast	Desloratadine
Prednisolone	RO-20-1724	Dipivefrin	Dipyridamole
				Prednisolone	Epinastine (Epinastine)	Dipivefrin	Limaprost
Prednisolone	Prostaglandin	Dipivefrin	Prostaglandin
				Prednisolone	Butylbenzene hydroxy acid	Dipivefrin	Butylbenzene hydroxy acid
Cyclosporin A	Brimonidine	Dipivefrin	Amoxicillin
				Cyclosporin A	Epinastine (Epinastine)	Dipivefrin	Amantadine
Cyclosporin A	Dipivefrin	Prostaglandin E	Ibudilast
				Cyclosporin A	Prostaglandin E	Misoprostol	Ibudilast
Cyclosporin A	Amantadine	Misoprostol	RO-20-1724
				Cyclosporin A	Acetaldine	Prostaglandin E	Dipyridamole
Cyclosporin A	Loratadine	Prostaglandin E	Rolipram
				Tacrolimus	Amantadine	Prostaglandin E	RO-20-1724
Dipyridamole	Amoxicillin	Limaprost	RO-20-1724
				Dipyridamole	Sertraline	Limaprost	Ibudilast
Loratadine	Nortriptyline	Thiochlorophenols	Nicarbazin
				Loratadine	Paroxetine	Dipyridamole	Rolipram
Amoxicillin	Butylbenzene hydroxy acid	Triamcinolone acetonide	Ibudilast
				Amoxicillin	Loratadine	Triamcinolone acetonide	Butylbenzene hydroxy acid
Amoxicillin	Nitrendipine

TABLE 1B

Medicine 1	Medicine 2	Medicine 1	Medicine 2
				Dexamethasone	Cyclosporin A	Dexamethasone	Tacrolimus
Fluorometholone	Cyclosporin A	Fluorometholone	Tacrolimus
				Hydrocortisone	Cyclosporin A	Hydrocortisone	Tacrolimus
Loteprednol etabonate	Cyclosporin A	Methoxypyr	Tacrolimus
				Methoxypyr	Cyclosporin A	Methylprednisolone	Tacrolimus
Methylprednisolone	Cyclosporin A	Prednisolone	Tacrolimus
				Prednisolone	Cyclosporin A	Prednisolone acetate	Tacrolimus

Prednisolone acetate	Cyclosporin A	Triamcinolone acetonide	Tacrolimus
				Rimexolone	Cyclosporin A
Triamcinolone acetonide	Cyclosporin A

In the methods, compositions, and kits of the invention, drugs having the same mechanism, structure, or therapeutic class may be used in place of one or more of the drugs listed in table 1A or table 1B. Table 2 provides exemplary categories.

TABLE 2

Species of	Medicine
		Alpha-2 adrenoceptor agonists	Brimonidine Acodeline
Anticoccidial compounds	Nicarbazin
		Antihistaminic agents	Loratadine epinastine
Infection resistance	Thiochlorophenols
		Anti-inflammatory	Butylbenzene hydroxy acid
Calcium channel blockers	Nitrendipine
		Corticosteroids	Prednisolone dexamethasone fluoromethalone hydrocortisone loteprednol etabonate Methylophilone methylprednisolone acetate prednisolone rimexolone triamcinolone
Electron donor-acceptor molecule complexes	Nicarbazin
		NMDA antagonists/anti-dyskinesia agents	Amantadine
NsIDI	Cyclosporin A tacrolimus

Phosphodiesterase inhibitors	Dipyridamole-ibudilast RO-20-1724 rolipram
		Prostaglandin	Prostaglandin E misoprostol limaprost
SSRI	Sertraline paroxetine
		Sympathomimetic agents	Dipivefrin
Tetra-substituted pyrimidopyrimidines	Dipyridamole
		Tricyclic compounds	Amoxicillin and nortriptyline

Thus, more broadly, the drug pairs in table 3 are useful in the methods, compositions, and kits of the present invention.

TABLE 3

Medicine 1	Medicine 2
		Antihistaminic agents	Phosphodiesterase inhibitors
Antihistaminic agents	SSRI
		Antihistaminic agents	Tricyclic compounds
Anti-infective agents	Anticoccidial compounds
		Corticosteroids	Antihistaminic agents
Corticosteroids	Phosphodiesterase inhibitors
		Corticosteroids	Prostaglandin
NsIDI	Alpha-2 adrenoceptor agonists
		NsIDI	Antihistaminic agents
NsIDI	NMDA antagonist/antagonistDyskinesia medicine
		NsIDI	Prostaglandin
NsIDI	Sympathomimetic agents
		Prostaglandin	Phosphodiesterase inhibitors

Prostaglandin	Tetra-substituted pyrimidopyrimidines
		Sympathomimetic agents	NMDA antagonists/anti-dyskinesia agents
Sympathomimetic agents	Prostaglandin
		Sympathomimetic agents	Tetra-substituted pyrimidopyrimidines
Sympathomimetic agents	Tricyclic compounds
		Tetra-substituted pyrimidopyrimidinesPyridine (I)	Phosphodiesterase inhibitors
Tetra-substituted pyrimidopyrimidines	SSRI
		Tetra-substituted pyrimidopyrimidines	Tricyclic compounds
Tricyclic compounds	Calcium channel blockers

These categories are discussed in more detail below.

Corticosteroids

In certain embodiments, corticosteroids may be used in the methods, compositions, or kits of the invention. Suitable corticosteroids include those of the selective glucocorticoid receptor agonist (SEGRA) class, 11 α,17 α, 21-trihydroxypregn-4-ene-3, 20-dione; 11 β,16 α,17, 21-tetrahydroxypregn-4-ene-3, 20-dione; 11 β,16 α,17, 21-tetrahydroxypregna-1, 4-diene-3, 20-dione; 11 β,17 α, 21-trihydroxy-6 α -methylpregn-4-ene-3, 20-dione; 11-dehydrocorticosterone; 11-deoxycorticosterone; 11-hydroxy-1, 4-androstadiene-3, 17-dione; 11-ketotestosterone; 14-hydroxyandrost-4-ene-3, 6, 17-trione; 15, 17-dihydroxyprogesterone; 16-methyl hydrocortisone; 17, 21-dihydroxy-16 α -methylpregna-1, 4, 9(11) -triene-3, 20-dione; 17 α -hydroxypregn-4-ene-3, 20-dione; 17 α -hydroxypregnenolic acid; 17-hydroxy-16 β -methyl-5 β -pregn-9 (11) -ene-3, 20-dione; 17-hydroxy-4, 6, 8(14) -gestriene-3, 20-dione; 17-hydroxypregna-4, 9(11) -diene-3, 20-dione; 18-hydroxycorticosterone; 18-hydroxy cortisone; 18-oxocortisol; 21-acetoxypregnenolone; 21-deoxyaldosterone; 21-deoxycorticosteron; 2-deoxyecdysone; 2-methyl cortisone; 3-dehydroecdysone; 4-pregnene-17 α,20 β, 21-triol-3, 11-dione; 6,17, 20-trihydroxypregn-4-en-3-one; 6 α -hydroxy cortisone; 6 alpha-methylprednisolone, 6 alpha-methylprednisolone 21-acetate, 6 alpha-methylprednisolone 21-hemisuccinate sodium salt, 6 beta-hydrocortisone, 6 alpha, 9 alpha-difluoroprednisolone 21-acetate 17-butyrate and 6-hydroxyanisolone; 6-hydroxydexamethasone; 6-hydroxy prednisolone; 9-flucortisone; abetasone dipropionate; aldosterone; alestrol; 1% hydrocortisone cream; amadone; amcinonide; anagesterone; androstenedione; anecortave acetate; beclomethasone; beclomethasone dipropionate; betamethasone 17-valerate; betamethasone sodium acetate; betamethasone sodium phosphate; betamethasone valerate; (ii) a dehydroepiandrosterone; budesonide; (ii) carpoterone; chlormadinone; prednisone; prednisone acetate; cholesterol; ciclesonide; clobetasol; clobetasol propionate; clobetasol; (ii) a chlorocortolone; clocortolone valerate; progesterone chloride; (ii) prednisolone; corticosterone; hydrocortisone; hydrocortisone acetate; hydrocortisone butyrate; hydrocortisone cypionate; hydrocortisone octanoate; hydrocortisone sodium phosphate; hydrocortisone sodium succinate; hydrocortisone valerate; cortisone; cortisone acetate; (ii) a kovar; trutodo pine; datura terpenol ketone; deflazacort, 21-deoxy hydrocortisone, dehydroepiandrosterone; demegestone; deoxycorticosterone; delosozone; a delphinidin derivative; (ii) donepezil; desoxybetamethasone; dexafen; dexamethasone; dexamethasone 21-acetate; dexamethasone acetate; dexamethasone sodium phosphate; dichloro pine; diflorasone; diflunisal diacetate; diflucortolone; difluprednate; dihydrobitter gourd bitter principle; a polyprednisole ester; a polypetaxole; ecdysone; ecdysterone; emoxolone; (ii) bentazone; glycyrrhetinic acid; fluzacort; the skin is relaxed; dichlorofluocinon; fludrocortisone; fludrocortisone acetate; fluoprogesterone; diflumetone; difluoromethylenedione pivalate; (ii) flumonade; flunisolide; fluocinolone acetonide; fluocinolone acetonide; fluocinonide; fluorocarbutyl ester; 9-flucortisone; fluocortolone; (ii) fluoro hydroxyandrostenedione; fluorometholone; fluorometholone acetate; fluoxymesterone; methyl fluridone acetate; fluprednidene; fluprednisolone; fluroxyprosol; fluticasone; fluticasone propionate; formylboxolone; 2, fulvestrant; formocortah; pregnenolone; glyderinine; halcinonide; halobetasol propionate; halometasone; prednisone halide; a haloprogesterone; a hydrocortisone ester; hydrocortisone cypionate; hydrocortisone; hydrocortisone 21-butyrate; hydrocortisone propionate acetate; hydrocortisone acetate; hydrocortisone butyrate propionate; hydrocortisone butyrate; hydrocortisone cypionate; hydrocortisone hemisuccinate; hydrocortisone probutate; hydrocortisone sodium phosphate; hydrocortisone sodium succinate; hydrocortisone valerate; hydroxyprogesterone; achyranthes bidentata sterone; isoflupredone; isophorone acetate; isoprednisone; loteprednol etabonate; (ii) meclosone; prednisolone pivalate; medroxyprogesterone; medroxyprogesterone; (ii) medroxyprogesterone; megestrol; megestrol acetate; mestranol; prednisone methyl; dehydromethyltestosterone; methylprednisolone; methylprednisolone aceponate; methylprednisolone acetate; methylprednisolone hemisuccinate; methylprednisolone sodium succinate; methyltestosterone; metrobolone; mometasone; mometasone furoate; mometasone furoate monohydrate; prednisone; nomegestrol; norgestimate; (ii) a ketene dimer; hydroxymethyltestosterone; p-flumethasone; p-flumethasone acetate; a ketosteroid; prednisolone ester; prednisolone ester; prednisolone; 21-diethylamino prednisolone acetate; 21-prednisolone hemisuccinate; prednisolone acetate; farnesylprednisolone; prednisolone hemisuccinate; 21- (β -D-glucuronide) prednisolone; prednisolone metasulphobenzoate; prednisolone sodium phosphate; prednisolone stearoyl glycolate; prednisolone tert-butylacetate; prednisolone tetrahydrophthalate; prednisone; prednisolone valerate; prednisolone; pregnenolone; pracinonide; (ii) trilonene; a progestin; progesterone; rhapontigenin; rimexolone; luoxibolone; amaranthone; stilzophyllin; alternative cortisone; tropanone; triamcinolone acetonide; triamcinolone acetonide; 21-triamcinolone acetonide palmitate; triamcinolone acetonide; triamcinolone acetonide; triamcinolone acetonide hexa; trimegestone; tacroline and wortmannin.

Tables 4A and 4B below provide standard recommended dosages for each steroid/disease combination.

TABLE 4A-recommended corticosteroid dosage of Standard

Table 4B-standard recommended corticosteroid dose for ocular administration

Ophthalmic corticosteroids	Minimum approved concentration for ophthalmic administration	Minimum standard recommended dose
			Clocotolone valerate	0.1％	N/A
Hydrocortisone	1.0％	0.5 μ g/day 3 times
			Dexamethasone	0.1％	0.05 μ g/day 4-6 times
Fluorometholone	0.1％	0.05 μ g/day 2-4 times
			Loteprednol etabonate	0.2％	0.1 μ g/4 times daily
Methoxypyr	1.0％	0.5. mu.g/up to 1 time per 4 hours
			Prednisolone acetate	0.12％	0.06 μ g/day 2-4 times
Rimexolone	1.0％	0.5 μ g/4 times daily

(N/A. not available)

Other standard recommended dosages for corticosteroids are provided, for example, in the diagnostic and therapeutic merck manual (17 th edition, MH Beers et al, merck) and in the Physicians' Desk Reference2003 (57 th edition, health economics officer et al, health economics limited, 2002). In one embodiment, the corticosteroid is administered at a dose equivalent to the dose of prednisolone as defined herein. For example, a low dose of corticosteroid can be considered to correspond to the dose of the low dose of prednisolone.

For ophthalmic administration, the corticosteroid can be administered at a concentration of 0.01-5% (e.g., 5.0%, 4.0%, 3.0%, 2.0%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

The low concentrations of the corticosteroids of the invention are 95% or less of the minimum concentrations approved. For example, a low concentration of a corticosteroid of the present invention can be 90%, 85%, 80%, 70%, 60%, 50%, 25%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% of the lowest concentration approved.

For ocular administration for example: low concentrations of clocortolone valerate, such as 0.01-0.1% (e.g., 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%), low concentrations of hydrocortisone, such as 0.01-1.0% (e.g., 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%), low concentrations of dexamethasone, such as 0.01-0.1% (e.g., 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%), low concentrations of fluorometholone, such as 0.01-0.1% (e.0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%), low concentrations of loteprednol carbonate, such as 0.01-0.01%, 0.0.1%, 0.09%, 0.0.0.0.06%, 0.0.0.0.0.01-0.05%, 0.0.0.0.06%, 0.0.0.01%, 0.0.0.0.0.06%, 0.0.0.01%, 0.0.0.06%, 0.0.0.0.0.01%, 0.0.06%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%), a low concentration of rimexolone of 0.01 to 1.0% (e.g., 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%), and a low concentration of prednisolone of 0.01 to 0.12% (e.g., 0.12%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, and 0.01%).

Steroid receptor modulators

In the methods, compositions, and kits of the invention, steroid receptor modulators (e.g., antagonists and agonists) may be used in place of or in addition to corticosteroids. Glucocorticoid receptor modulators useful in the methods, compositions, and kits of the present invention include compounds described in the following patents: U.S. patent nos. 6,380,207, 6,380,223, 6,448,405, 6,506,766 and 6,570,020, U.S. patent application publication nos. 2003/0176478, 2003/0171585, 2003/0120081, 2003/0073703, 2002/015631, 2002/0147336, 2002/0107235, 2002/0103217 and 2001/0041802, and PCT publication No. WO00/66522, each of which is incorporated herein by reference. Other steroid receptor modulators may also be used in the methods, compositions and kits of the present invention as described in the following patents: U.S. patent nos. 6,093,821, 6,121,450, 5,994,544, 5,696,133, 5,696,127, 5,693,647, 5,693,646, 5,688,810, 5,688,808 and 5,696,130, each of which is incorporated herein by reference.

Other Compounds

In addition to the pharmaceutical combinations of table 1A or table 3, other compounds useful in the methods, compositions, and kits of the present invention are: a-348441(Karo Bio), adrenocortical extract (GlaxoSmithKline), Asaktide (Aventis), ambcort (Schering AG), Allometasone (Taisho), ATSA (Pfizer), bitolterol (Elan), CBP-2011(Inkine pharmaceutical), sibutran (Novartis), CGP-13774(Kissei), ciclesonide (Altana), cicletasone (Aventis), clobetasol butyrate (Glaxo Smithline), prednisolone (Hoffmann-Roche), collismycin A (Kirrin cucurbit), picrin E (PfNIH), deflazacort (Aventis), deferiprone propionate (SSP), dexamethasone acetate (Schering-Plough), dexamethasone acetate (Glaxomolyton oleate (Glaxon), dexamethasone acetate (Abutin), dexamethasone acetate (Vizanol), and flunopyrne acetate (Avermectin (La), and fluvictorin (Avenun) esters (Avermectin (Roffman), and dexamethasone (Vizanol), and dexamethasone acetate (Avenun acetate), and dexamethasone (Viffman) esters (Viffonine), and (Viffonine esters (Viffman) and dexamethasone acetate (Viffman) and (Viffoni) and (Viffman) esters (Viffoni) and (Viffman) and (Vi, Butyl Fluorococadine (Schering AG), Fluorococalone monohydrate (Schering AG), GR-250495X (Glaxo Smith Kline), Halometasone (Novartis), Halopropion (Dainippon), HYC-141(Fidia), icomethasone enbutate (Hovione), Irvinolide (AstraZeneca), L-6485(Vicuron), Lipocort (Draxis Healthy), Closene (Aventis), Methylon (Schering-Plough), Nafelone (Bril-Myers quiSbb), NCX-1015(NicOx), NCX-1020(NicOx), NCX-1022(NicOx), Nicohnuchi (Yamanouchi), NIK-236 (Nikkenmills), NS-SSP (SSP-2766), Orbestrol (Ak-11156), Oryza-36663 (Ak), Avonene-3683 (Avonedar-26559), Avonedare-r-26559 (Avonedare), Avonene 26559 (Avonedare-3), Avonedare-26559 (Avonedare), Narone), Naphthori-3 (R-26559), and Ak-3 (Avonedare), and Alkone) Sch-19457(Schering-Plough), T25(Matrix Therapeutics), TBI-PAB (Sigma-Tau), ticarberre propionate (Hoffmann-La Roche), temofluo (Solvay), temoberarus (Hoffmann-La Roche), TSC-5(Takeda), ZK-73634(Schering AG), antibiotics (minocycline, penicillin, cephalosporin, tetracycline, oxytetracycline, chlortetracycline, metronidazole, chloramphenicol, streptomycin, neomycin, sulfonamides, phenolic compounds, quaternary ammonium compounds, doxycycline, preservatives (e.g., chlorhexidine), tranexamic acid, allantoin, epsilon-aminocaproic acid, lysozyme, dihydrocholesterol, beta-glycyrrhetinic acid, platelet aggregation inhibitors (e.g., abciximab, aspirin, cilostazol, clopidogrel, dipyridamole, eptid, lopeptid, lopamisole, lopamitradine or tirofiban), Anticoagulants (e.g., dalteparin, danaparoid, enoxaparin, heparin, tinzaparin, or warfarin), antipyretics (e.g., acetaminophen), ticlopidine, clopidogrel, angiotensin converting enzyme inhibitors, beta blockers, pentoxifylline, cilostazol, estrogen replacement therapy, and lipid lowering drugs (e.g., cholestyramine, colestipol, nicotinic acid, gemfibrozil, probucol, ezetimibe, or statins such as atorvastatin, rosuvastatin, lovastatin, simvastatin, pravastatin, cerivastatin, and fluvastatin). These agents may be administered simultaneously or within 14 days of the methods of the invention. If desired, one or more of the foregoing agents may be co-formulated with one or more of the agents of the present invention to form a composition.

Non-steroidal immunophilin-dependent immunosuppressants

In one embodiment, the invention features methods, compositions, and kits for using a non-steroidal immunophilin-dependent immunosuppressant (NsIDI).

In healthy individuals, the immune system targets infectious microorganisms and abnormal cell types using cellular effectors such as B cells and T cells when normal cells are left intact. In individuals with autoimmune disease or transplanted organs, activated T cells damage healthy tissue. Calcineurin inhibitors (such as cyclosporine, tacrolimus, pimecrolimus, ABT-281, ISAtx247) and rapamycin target many types of immunoregulatory cells, including T cells, and suppress immune responses in organ transplants and autoimmune diseases.

In one embodiment, the NsIDI is cyclosporin A and is administered in an amount of 0.05 to 50 mg/Kg/day (e.g., orally in an amount of 0.1 to 12 mg/Kg/day). In another embodiment, the NsIDI is cyclosporine administered as a 0.05% ophthalmic emulsion twice daily. In another embodiment, the NsIDI is tacrolimus and is administered in an amount of 0.0001 to 20 mg/Kg/day (e.g., orally in an amount of 0.01 to 0.2 mg/Kg/day). In another embodiment, the NsIDI is tacrolimus and is administered as a 0.02% ophthalmic suspension. In another embodiment, the NsIDI is rapamycin and is administered in an amount from 0.1 to 502 mg/day (e.g., as a single drug dose of 6 mg/day followed by a maintenance dose of 2 mg/day). In another embodiment, the NsIDI is everolimus and is administered at a dose of 0.75-8 mg/day. In yet another embodiment, the NsIDI is pimecrolimus, administered in an amount of 0.1-200 mg/day (e.g., twice daily in 1% ointment for atopic dermatitis or 60mg a day for psoriasis), or the NsIDI is a calcineurin-binding peptide, administered in an amount and at a frequency sufficient to treat the patient. Two or more NsIDI may be administered simultaneously.

Cyclosporin

Cyclosporine is a fungal metabolite, comprising a class of cyclic oligopeptides, used as immunosuppressive agents. Cyclosporin A is a hydrophobic cyclic polypeptide consisting of 11And (4) amino acid. It binds to the intracellular receptor cyclophilin, forming a complex. The cyclosporin/cyclophilin complex binds to and inhibits calcineurin, Ca²⁺Calmodulin-dependent serine-threonine-specific protein phosphatases. The signal transduction events required for the mediation of T-Cell activation by neurotalcin (reviewed in Schreiber et al, Cell 70: 365-. Cyclosporine and functional and structural analogs thereof inhibit T cell-dependent immune responses by inhibiting antigen-triggered signal transduction. This inhibition reduces the expression of proinflammatory cytokines such as IL-2.

Many different cyclosporins (e.g., cyclosporins A, B, C, D, E, F, G, H and I) are produced by fungi. Cyclosporin a is available from Novartis under the trade name NEORAL. Structural and functional analogs of cyclosporin a include cyclosporines having one or more fluorinated amino acids (as described in U.S. patent No. 5,227,467); cyclosporines with modified amino acids (as described in U.S. patent nos. 5,122,511 and 4,798,823); and deuterated cyclosporines such as ISAtx247 (described in U.S. patent application publication No. 2002/0132763a 1). Additional cyclosporine analogs are described in U.S. patent nos. 6,136,357, 4,384,996, 5,284,826 and 5,709,797. Cyclosporine analogs include, but are not limited to: D-Sar (alpha-SMe)³Valine²DH-Cs (209-825), Allo-Thr-2-Cs, norvaline-2-Cs, D-alanine (3-acetylamino) -8-Cs, threonine-2-Cs and D-MeSer-3-Cs, D-serine (O-CH)₂CH₂-OH) -8-Cs and D-serine-8-Cs, as described in Cruz et al (Antiicrob. AgentsChemother.44: 143-149, 2000) describe these cyclosporin analogs.

Cyclosporine is highly hydrophobic and readily precipitates in the presence of water (e.g., upon contact with body fluids). In U.S. Pat. nos.4,388,307, 6,468,968, 5,051,402, 5,342,625, 5,977,066 and 6,022,852, methods are described that provide cyclosporin formulations with improved bioavailability. In U.S. patent nos. 5,866,159, 5,916,589, 5,962,014, 5,962,017, 6,007,840 and 6,024,978, compositions of cyclosporine microemulsions are described.

Cyclosporin may be administered intravenously or orally, but oral administration is preferred. To overcome the hydrophobicity of cyclosporin a, intravenous cyclosporin a may be provided in an ethanolic-polyoxyethylated castor oil vehicle, which must be diluted prior to administration. Cyclosporin a may be provided, for example, as a 25mg microemulsion or 100mg tablet, 100mg/ml oral solution (NEORAL), or 0.05% ophthalmic emulsion.

Typically, the patient dose of oral cyclosporin will vary depending on the patient's condition, but some standard recommended doses are provided herein. Patients undergoing organ transplantation typically receive an initial dose of oral cyclosporin A in an amount of 12-15 mg/kg/day. The dose was then gradually reduced at 5%/week until a maintenance dose of 7-12 mg/kg/day was reached. Most patients are preferably given 2-6 mg/kg/day intravenously. For patients diagnosed with Crohn's disease or ulcerative colitis, a dose of 6-8 mg/kg/day is typically administered. For patients diagnosed with systemic lupus erythematosus, doses of 2.2-6.0 mg/kg/day are typically administered. For psoriasis or rheumatoid arthritis, a typical dose is 0.5-4 mg/kg/day. Table 5 shows the recommended dosing regimen. Other useful dosages include 0.5-5 mg/kg/day, 5-10 mg/kg/day, 10-15 mg/kg/day, 15-20 mg/kg/day, or 20-25 mg/kg/day.

TABLE 5

Compound (I)

Ectopic skinInflammation of the stomach

Psoriasis disease

RA

Segmental enteritis

UC

Transplantation

SLE

CsA (NEORAL)

N/A

0.5-4 mg/kg/day

0.5-4 mg/kg/day

6-8 mg/kg/day (oral fistulation)

6-8 mg/kg/day (oral)

About 7 to 12 mg/kg/day

2.2-6.0 mg/kg/day

Tacrolimus

0.03-0.1% ointment/twice a day (30 and 60 g tube)

0.05-1.15 mg/kg/day (oral)

1-3 mg/day (oral)

0.1-0.2 mg/kg/day (oral)

0.1-0.2 mg/kg/day (oral)

0.1-0.2 mg/kg/day (oral)

N/A

Pimecrolimus

1% ointment/twice daily (15, 30, 100 g tube)

40-60 mg/day (oral cavity)Clothes)

40-60 mg/day (oral)

80-160 mg/day (oral)

160-240 mg/day (oral)

40-120 mg/day (oral)

40-120 mg/day (oral)

CsA ═ cyclosporin A

Rheumatoid Arthritis (RA)

UC ═ ulcerative colitis

SLE ═ systemic lupus erythematosus

The minimum approved ophthalmic concentration of cyclosporin a was 0.05%. Low concentrations of cyclosporin a are 0.04%, or more preferably 0.03%, 0.02%, 0.01%, 0.008%, 0.005% or 0.001%. The lowest standard recommended ophthalmic dose of cyclosporin a is 0.2 μ g twice daily.

Tacrolimus

Tacrolimus (FK506) is an immunosuppressant targeting the T intracellular signal transduction pathway. Tacrolimus binds to the intracellular protein FK506 binding protein (FKBP-12), which FKBP-12 is structurally unrelated to cyclophilin. The FKBP/FK506 complex binds to calcineurin, inhibiting the phosphatase activity of calcineurin. This inhibition prevents dephosphorylation and nuclear translocation of nuclear factor of activated T cells (NFAT), which is a nuclear component that initiates the production of pro-inflammatory cytokines (e.g., IL-2, interferon gamma) and transcription of genes required for T cell activation. Thus, tacrolimus inhibits activation of T cells.

Tacrolimus is a macrolide antibiotic produced by Streptomyces tsukubaensis (Streptomyces tsukubaensis) in the mountainous soil. It suppresses the immune system and prolongs the survival of transplanted organs. It is currently used in oral formulations and injections. Tacrolimus capsules contain 0.5mg, 1mg or 5mg of anhydrous tacrolimus with a gelatin capsule shell. The injection comprised 5mg of tacrolimus anhydrous in castor oil and alcohol, diluted with 0.9% sodium chloride or 5% glucose prior to injection. Although oral administration is preferred, patients who cannot orally administer the capsule may receive injectable tacrolimus. The initial dose should be given by continuous intravenous infusion immediately 6 hours after transplantation.

Tanaka et al (j.am. chem. soc, 109: 5031, 1987) and in us patent nos.4,894,366, 4,929,611 and 4,956,352 describe tacrolimus and tacrolimus analogs. In U.S. Pat. No. 5,254,562, FK 506-related compounds are described, including FR-900520, FR-900523 and FR-900525; in U.S. Pat. Nos. 5,250,678, 532,248, 5,693,648, O-aryl, O-alkyl, O-alkenyl and O-alkynyl macrolides are described; in U.S. Pat. No. 5,262,533, amino O-aryl macrolides are described; in U.S. Pat. No. 5,284,840, alkylene macrolides are described; in U.S. Pat. No. 5,208,241, N-heteroaryl, N-alkylheteroaryl, N-alkenylheteroaryl and N-alkynylheteroaryl macrolides are described; in us patent No. 5,208,228, amino macrolides and their derivatives are described; in us patent No. 5,189,042, fluoro macrolides are described; in U.S. Pat. No. 5,162,334, amino O-alkyl, O-alkenyl and O-alkynyl macrolides are described; in U.S. Pat. No. 5,143,918, halogenated macrolides are described.

Although the recommended dose varies depending on the condition of the patient, the standard recommended dose is provided below. Typical patients diagnosed with Crohn's disease or ulcerative colitis have tacrolimus administered orally at 0.1-0.2 mg/kg/day. Typical doses of oral tacrolimus for patients undergoing organ transplantation are 0.1-0.2 mg/kg/day. Patients for rheumatoid arthritis typically receive 1-3 mg/day of tacrolimus orally. For the treatment of psoriasis, the patient takes tacrolimus 0.01-0.15 mg/kg/day orally. Atopic dermatitis can be treated by applying 0.03-0.1% tacrolimus ointment twice daily to the affected site. Patients taking tacrolimus capsules orally typically receive the first dose immediately 6 hours after transplantation or 8-12 hours after intermittent intravenous infusion of tacrolimus. Other suggested tacrolimus dosages include 0.005-0.01 mg/kg/day, 0.01-0.03 mg/kg/day, 0.03-0.05 mg/kg/day, 0.05-0.07 mg/kg/day, 0.07-0.10 mg/kg/day, 0.10-0.25 mg/kg/day, or 0.25-0.5 mg/kg/day.

Tacrolimus is widely metabolized by mixed-function oxidase systems, particularly by the cytochrome P-450 system. The main mechanisms of metabolism are demethylation and hydroxylation. Although various tacrolimus metabolites may exhibit immunosuppressive biological activity, 13-demethylation metabolites have been reported to have the same activity as tacrolimus.

Pimecrolimus

Pimecrolimus is a 33-epichloro derivative of ascomycin macrolactam (macrolactim). Structural and functional analogs of pimecrolimus are described in U.S. patent No. 6,384,073. Pimecrolimus is particularly useful for the treatment of atopic dermatitis. Currently 1% pimecrolimus ointment is used. Table 5 shows the recommended dosing regimen for pimecrolimus. Although individual dosages will vary depending on the patient's condition, some standard recommended dosages are provided below. Can be used for treating psoriasis or rheumatoid arthritis by oral administration of pimecrolimus at a dose of 40-60 mg/day. Pimecrolimus can be administered at a dose of 80-160 mg/day to treat Crohn's disease or ulcerative colitis. 160-240 mg/day pimecrolimus may be administered to a patient undergoing organ transplantation. Pimecrolimus at 40-120 mg/day may be administered to a patient diagnosed with systemic lupus erythematosus. Other useful dosages of pimecrolimus include 0.5-5 mg/day, 5-10 mg/day, 10-30 mg/day, 40-80 mg/day, 80-120 mg/day or even 120-200 mg/day.

Rapamycin

Rapamycin is a cyclic lactone produced by Streptomyces hygroscopicus (Streptomyces hygroscopicus). Rapamycin is an immunosuppressant, inhibiting the activation and proliferation of T cells. Like cyclosporin and tacrolimus, rapamycin forms a complex with immunophilin FKBP-12, but the rapamycin-FKBP-12 complex does not inhibit the phosphatase activity of calcineurin. The rapamycin immunophilin complex binds to and inhibits the mammalian kinase target of rapamycin (mTOR). mTOR is a kinase required for cell cycle progression. Inhibition of mTOR kinase activity blocks T cell activation and secretion of pro-inflammatory cytokines.

Structural and functional analogs of rapamycin include monoacylated rapamycin derivatives and diacylated rapamycin derivatives (U.S. Pat. No. 4,316,885); water-soluble prodrugs of rapamycin (U.S. Pat. No. 4,650,803); carboxylic acid esters (PCT publication No. WO 92/05179); carbamates (U.S. Pat. No. 5,118,678); amide esters (U.S. Pat. No. 5,118,678); biotin esters (U.S. Pat. No. 5,504,091); fluorinated esters (U.S. Pat. No. 5,100,883); acetals (U.S. Pat. No. 5,151,413); silyl ethers (U.S. Pat. No. 5,120,842); bicyclic derivatives (U.S. patent No. 5,120,725); rapamycin dimer (U.S. Pat. No. 5,120,727); o-aryl, O-alkyl, O-alkenyl, and O-alkynyl derivatives (U.S. Pat. No. 5,258,389); and deuterated rapamycin (U.S. patent No. 6,503,921). Additional rapamycin analogs are described in U.S. patent nos. 5,202,332 and 5,169,851.

Oral rapamycin is currently available in liquid formulations and tablets. Rapamycin liquid at 1mg/ml was diluted in water or orange juice prior to administration. Tablets containing 1 or 2mg of rapamycin may also be used. Rapamycin is preferably administered once daily as soon as possible after transplantation. Can be absorbed quickly and completely after being taken orally. Typically, the rapamycin dosage for a patient varies depending on the condition of the patient, but some standard recommended dosages are provided below. The initial drug loading dose of rapamycin was 6 mg. Typical maintenance doses are then 0.5-2 mg/day. Alternatively, a drug loading dose of 3mg, 5mg, 10mg, 15mg, 20mg or 25mg and a maintenance dose of 1mg, 3mg, 5mg, 7mg or 10 mg/day may be used. In patients weighing less than 40kg, typically depending on body surface areaAdjusting the dose of rapamycin; generally 3mg/m is used²Drug loading dose per day and 1mg/m²Maintenance dose/day.

Peptide moieties

Natural, synthetic or chemically modified peptides, peptide analogs, peptide fragments, which impair calcineurin-mediated dephosphorylation and nuclear translocation of NFAT, are suitable for practicing the present invention. For example, by Aramburu et al, Science 285: 2129-2133, 1999) and Aramburu et al, mol.cell 1: 627-637, 1998) describe examples of peptides that act as calcineurin inhibitors by inhibiting activation of NFAT and NFAT transcription factors. These agents are a class of calcineurin inhibitors and are useful in the methods, compositions, and kits of the present invention.

Phosphodiesterase inhibitors

In certain embodiments, phosphodiesterase inhibitors can be used in the methods, compositions, and kits of the invention. Suitable phosphodiesterase inhibitors include phosphodiesterase type III (cAMP-specific-cGMP inhibitable form) inhibitors, phosphodiesterase type IV (high affinity-high specific cAMP type) inhibitors and phosphodiesterase type V (cGMP specific form) inhibitors. Examples of type III phosphodiesterase inhibitors include bipyridines such as mihinone and aminopyridone, imidazolones such as piroximone and enoximone, dihydropyridazinones such as imazodan, 5-methylimidazolidan, indolidan and ICI 118233(6- (p- (3-methylureido) phenyl) -3(2H) -pyridazinone), quinolinone compounds such as cilostamide, cilostazol and vinrinone and other compounds such as bemoradan, anergrelide, cyguanzodan, quqinoline, pikendan, SKF-94120(5- (4-acetamidophenyl) pyrazin-2- (1H) -one), SKF-95654, lixazinone and isomazole. Examples of phosphodiesterase type IV inhibitors include rolipram and rolipram derivatives such as RO-20-1724(4- (3-butoxy-4-methoxyphenyl) -imidazolidinone), nitroquinazone and nitroquinazone derivatives such as CP-77059(1- (methoxycarbonylphenyl) -3-benzylpyrido [2, 3-d ] pyrimidine-2, 4(1H, 3H) dione) and RS-25344-00(1- (3-nitrophenyl) -3- (4-pyridylmethyl) -1, 2,3, 4-tetrahydropyrido (2, 3-d) pyrimidine-2, 4-dione)), xanthine derivatives such as denbufylline and ICI63197 and various other compounds such as EMD54622(5- [1- (3, 4-dimethoxybenzoyl) -4, 4-dimethyl-1, 2,3, 4-tetrahydroquinolin-6-yl ] -6-methyl-3, 6-dihydro-1, 3, 4-metiram-2-one), LAS-31025 (1-propyl-3- (4-chlorophenyl) xanthine; also known as arophylline) and etazolate. Examples of phosphodiesterase type V inhibitors include zaprinast, MY5445(N- (3-chlorophenyl) -4-phenyl-1-phthalazinamine), dipyridamole and sildenafil. In PCT publication Nos. WO 94/28902 and WO 96/16644, sildenafil (5- [ 2-ethoxy-5- (4-methyl-1-piperazinylsulfonyl) -phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one) and other suitable phosphodiesterase type V inhibitors (e.g., 5- (2-ethoxy-5-morpholinoacetyl-phenyl) -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- (5-morpholino-acetyl-2-n-propoxyphenyl) -1- Methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- [ 2-ethoxy-5- (4-methyl-1-piperazinylsulfonyl) -phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- [ 2-allyloxy-5- (4-methyl-1-piperazinylsulfonyl) -phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, and pharmaceutically acceptable salts thereof, 5- [ 2-ethoxy-5- [4- (2-propyl) -1-piperazinylsulfonyl) -phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- [ 2-ethoxy-5- [4- (2-hydroxyethyl) -1-piperazinylsulfonyl) -phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- [5- [4- (2-hydroxyethyl) -1-piperazinylsulfonyl ] -2-n-propoxyphenyl ] -1-methyl-3 -n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- [ 2-ethoxy-5- (4-methyl-1-piperazinylcarbonyl) phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 5- [ 2-ethoxy-5- (1-methyl-2-imidazolyl) phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, 1, 3-dimethyl-5-benzylpyrazolo [4, 3-d ] pyrimidin-7-one, 2- (2-propoxyphenyl) -6-purinone, 6- (2-propoxyphenyl) -1, 2-dihydro-2-oxopyridine-3-carboxamide, 2- (2-propoxyphenyl) -pyrido [2, 3-d ] pyrimidin-4 (3H) -one, 7-methylsulfanyl-4-oxo-2- (2-propoxyphenyl) -3, 4-dihydro-pyrimido [4, 5-d ] pyrimidine, 6-hydroxy-2 (2-propoxyphenyl) -pyrimidine-4-carboxamide, 1-ethyl-3-methylimidazo [1, 5a ] quinoxalin-4 (5H) -one, 4-phenyl-methylamino-6-chloro-2- (1-imidazolyl) quinazoline, 5-ethyl-8- [3- (N-cyclohexyl-N-methyl-carbamoyl) -propyloxy ] -4, 5-dihydro-4-oxo-pyrido [3, 2-e ] -pyrrolo [1, 2-a ] pyrazine, 5 ' -methyl-3 ' - (phenylmethyl) -spiro [ cyclopentane-1, 7 ' (8 ' H) - (3 ' H) -imidazo [2, 1-b ] purine ]4 ' (5 ' H) -one, 1- [ 6-chloro-4- (3, 4-methylenedioxybenzyl) -aminoquinazolin-2-yl) piperidine-4-carboxylic acid, (6R, 9S) -2- (4-trifluoromethyl-phenyl) methyl-5-methyl-3, 4,5, 6a, 7,8, 9,9 a-octahydrocyclopenta (cyclopent) [4, 5] -imidazo [2, 1-b ] -purin-4-one, 1-tert-butyl-3-phenylmethyl-6- (4-pyridyl) pyrazolo [3, 4-d ] -pyrimidin-4-one, 1-cyclopentyl-3-methyl-6- (4-pyridyl) -4, 5-dihydro-1H-pyrazolo [3, 4-d ] pyrimidin-4-one, 2-butyl-1- (2-chlorophenylmethyl) 6-ethoxy-carbonylbenzimidazole, 2- (4-carboxypiperidino) -4- (3, 4-methylenedioxybenzyl) amino-6-nitroquinazoline, and 2-phenyl-8-ethoxycyclohepta (cyclohexpt) imidazole. Still other phosphodiesterase type V inhibitors useful in conjunction with the present invention are IC-351(ICOS), also known as tadalafil, 4-bromo-5- (pyridylmethylamino) -6- [3- (4-chlorophenyl) -propoxy ] -3(2H) pyridazinone, 1- [4- [ (1, 3-benzodioxol-5-ylmethyl) amino ] -6-chloro-2-quinazolinyl ] -4-piperidine-carboxylic acid, monosodium salt, (+) -cis-5, 6a, 7, 9,9, 9 a-hexahydro-2- [4- (trifluoromethyl) -phenylmethyl-5-methyl-cyclopenta-4, 5] imidazo [2, 1-b ] purin-4 (3H) one, furloxacillin, cis-2-hexyl-5-methyl-3, 4,5, 6a, 7,8, 9,9 a-octahydrocyclopenta [4, 5] imidazo [2, 1-b ] purin-4-one, 3-acetyl-1- (2-chlorobenzyl) -2-propylindole-6-carboxylate, 4-bromo-5- (3-pyridylmethylamino) -6- (3- (4-chlorophenyl) -propoxy) -3- (2H) pyridazinone, 1-methyl-5- (5-morpholinoacetyl-2-n-propoxyphenyl) -3-n-propyl-1, 6-dihydro-7H-pyrazolo (4, 3-d) pyrimidin-7-one, 1- [4- [ (1, 3-benzodioxol-5-ylmethyl) amino ] -6-chloro-2-quinazolinyl ] -4-piperidinecarboxylic acid, monosodium salt, Pharmaprojes No. 4516 (GlaxoWellcome), Pharmaprojes No. 5051 (Bayer), Pharmaprojes No. 5064 (Kyowa Hakko; see WO 96/26940), Pharmaprojes No. 5069 (Schering Plough) and Sch-51866. Other phosphodiesterase inhibitors are disclosed in U.S. patent No. 6,469,016.

Other phosphodiesterase inhibitors that may be used in the present invention include fexofenadine, pyraclostrobin, Org 20241, MCI-154, roflumilast, toliprone, mibefradil, pyrazolopyrimidinones (such as those disclosed in WO 98/49166), motapine, pidan, zadaparine, cyguanodazodan, CI 930, EMD 53998, imazodan, sarrinone, loprinone hydrochloride, 3-pyridinecarbonitrile derivatives, abiphylline, tobafylline, doxofylline, chophylline, pentoxyline, nanrinone, cilostazol, cyclohexolamide, MS 857, piroximone, milrinone, aminopyridone, tolafenac, papaferin, E4021, thienopyrimidine derivatives (such as those disclosed in PCT publication No. WO 98/17668), trifluoroacetic acid, tetrahydropiperazino [1, 2-carbolino ] beta 1-1, 4-diketone derivatives (such as those disclosed in U.S. Pat. No. 5,859,006 and PCT publications WO 97/03985 and WO 97/03675), carboline derivatives (such as those disclosed in PCT publication WO 97/43287), 2-pyrazolin-5-one derivatives (such as those disclosed in U.S. Pat. No. 5,869,516), fused pyridazine derivatives (such as those disclosed in U.S. Pat. No. 5,849,741), quinazoline derivatives (such as those disclosed in U.S. Pat. No. 5,614,627), anthranilic acid derivatives (such as those disclosed in U.S. Pat. No. 5,714,993), and imidazoquinazoline derivatives (such as those disclosed in PCT publication WO 96/26940). Also included are phosphodiesterase inhibitors disclosed in U.S. patent nos. 3,850,941, 4,097,483, 4,193,926, 4,578,392, 4,925,849, 4,994,453 and 5,296,490, and PCT publication nos. WO 99/21562 and WO 99/30697.

Alpha-2 adrenergic agonists

Alpha-2 adrenergic agonists can be used in the methods, compositions, and kits of the invention. An exemplary alpha-2 adrenergic agonist is brimonidine (5-bromo-N- (4, 5-dihydro-1H-imidazol-2-yl) -6-quinoxalinamine) described in U.S. Pat. No. 3,890,319. Other alpha-2 adrenergic agonists that can be used in the methods, compositions, and kits of the invention include apraclonidine, clonidine, dexmedetomidine, guanabenz, guanfacine, medetomidine, methyldopa, oxymetazoline, tizanidine, and (±) - (R, S) -5, 6-diisobutyloxy-2-methylaminotetralin. Other alpha-2 adrenergic agonists are described in the following patents: U.S. Pat. No. 5

2,868,818, respectively; 3,158,648, respectively; 3,202,660, respectively; 3,632,645, respectively; 3,843,668, respectively; 3,890,319, respectively; 4,029,792, respectively; 4,486,432, respectively; 4,517,199, respectively; 4,576,954, respectively; 4,910,214, respectively; 5,021,410, 5,037,829; 5,077,292, 5,091,528; 5,112,822, respectively; 5,130,441, respectively; 5,180,721, respectively; 5,198,442, respectively; 5,204,347, respectively; 5,215,991, respectively; 5,231,096, respectively; 5,237,072, respectively; 5,252,595, respectively; 5,281,591, respectively; 5,300,504, respectively; 5,326,763, respectively; 5,373,010, respectively; 5,418,234, respectively; 5,478,858, respectively; 5,541,210, respectively; 5,552,403, respectively; 5,561,132, respectively; 5,576,437, respectively; 5,578,607, respectively; 5,580,892, respectively; 5,587,376, respectively; 5,684,156, respectively; 5,691,370, respectively; 5,703,077; 5,708,015, respectively; 5,714,966, respectively; 5,739,148, respectively; 5,756,503, respectively; 5,773,440, respectively; 5,804,587, respectively; 5,834,470, respectively; 5,856,329, respectively; 5,914,342, respectively; 5,916,900, respectively; 5,965,595, respectively; 6,066,740, respectively; 6,110,952, respectively; 6,117,871, respectively; 6,162,818, respectively; 6,172,095, respectively; 6,194,415, respectively; 6,225,331, respectively; 6,242,442, respectively; 6,248,741, respectively; 6,294,563, respectively; 6,306,877, respectively; 6,316,441, respectively; 6,316,637, respectively; 6,323,204; 6,391,878, respectively; 6,395,764, respectively; 6,403,626, respectively; 6,423,724, respectively; 6,436,978, respectively; 6,436,982, respectively; 6,465,464, respectively; 6,486,190, respectively; 6,495,583, respectively; 6,562,873, respectively; 6,627,210, respectively; 6,641,834, respectively; 6,673,337, respectively; and 6,953,813.

For ophthalmic use, brimonidine is available as a 0.2% brimonidine tartrate ophthalmic solution.

Prostaglandin

Prostaglandins may be used in the methods, compositions, and kits of the present invention. The prostaglandins include prostaglandin E1, dinoprostone, misoprostol, limaprost, bimatoprost, travoprost, unoprostone, latanoprost, prostaglandin E2, prostaglandin a1, prostaglandin a2, prostaglandin B1, prostaglandin B2, prostaglandin D2, prostaglandin F1 α, prostaglandin F2 α, prostaglandin I1, prostaglandin-ici 74205, prostaglandin F2 β, 6-keto-prostaglandin F1 α, prostaglandin E1 ethyl ester, prostaglandin E1 methyl ester, prostaglandin F2 methyl ester, abaprost, ornoprost, 13, 14-dihydro prostaglandin F2 α and prostaglandin J. Ophthalmic formulations of prostaglandins include 0.03% bimatoprost, 0.004% travoprost, 0.15% unoprostone, and 0.005% latanoprost.

Tetra-substituted pyrimidopyrimidines

In certain embodiments, a tetra-substituted pyrimidopyrimidine or adenosine activity up-regulator and a second pharmaceutical agent may be used in combination in the methods, compositions, and kits of the present invention. "tetra-substituted pyrimidopyrimidines" refer to compounds having the formula (V):

wherein each Z and each Z' are independently N, O, C,Or

When Z or Z' is O orWhen Z or Z' is N, then p is 1,OrWhen Z or Z 'is C, then p is 2, and when Z or Z' is C, then p is 3. In the formula (V), each R₁Independently X, OH, N-alkyl (wherein the alkyl group has 1 to 20, more preferably 1 to 5 carbon atoms); branched or unbranched alkyl groups having 1 to 20, more preferably 1 to 5 carbon atoms; or heterocyclic, preferably as defined for formula (V). Or, when p > 1, two R of a common Z or Z' atom₁The radicals in combination may represent- (CY)₂)_k-, where k is an integer of 4 to 6 (including 4 and 6). Each X is independently Y, CY₃、C(CY₃)₃、CY₂CY₃、(CY₂)_1-5OY, Structure C_nY_2n-1Wherein n-3-7 (including 3 and 7). Each Y is independently H, F, Cl, Br or I. In one embodiment, each Z is the same moiety, each Z 'is the same moiety, and Z' are different moieties.

"adenosine activity up-regulator" refers to adenosine and any compound that mimics or enhances the physiological effects of adenosine, such as adenosine receptor agonists, adenosine transport inhibitors, adenosine kinase inhibitors, and Phosphodiesterase (PDE) inhibitors, as described herein.

Adenosine receptor agonists

Adenosine receptor agonists include adenosine hemisulfate, adenosylamine solids, N⁶- (4-amino-3-iodophenyl) methyl-5' -N-methylcarboxyamidoadenosine (I-AB-MECA); n- ((2-methylphenyl) methyl) adenosine (adenyldil); 2- (1-hexynyl) -N-methyladenosine (HEMADO); n- (1-methyl-2-phenylethyl) adenosine (R-PIA); n is a radical of⁶- (R-4-hydroxyphenylisopropyl) adenosine (HPIA); n is a radical of⁶-Cyclopentyladenosine (CPA); n is a radical of⁶-cyclopentyl-2- (3-phenylaminocarbonyltriazen-1-yl) adenosine (TCPA); n- ((1S, trans) -2-hydroxycyclopentyl) adenosine (GR 79236); n is a radical of⁶-cyclohexyl adenosine (CHA); 2-chloro-N⁶-cyclopentyladenosine (CCPA); n-ethylcarboxamidoadenosine (NECA); 2- (4- (2-carboxyethyl) phenethylamino) -5' -N-ethylformylAminoadenosine (CGS 21680); n is a radical of⁶- (3-iodobenzyl) -5' -N-methylcarboxyamidoadenosine (IB-MECA); 2- (cyclohexylmethylenehydrazino) adenosine (WRC 0470); 2- (4- (2-carboxyethyl) phenethylamino) -5' -N-ethylcarboxamidoadenosine (CGS 21680); n is a radical of⁶- (2- (3, 5-dimethoxyphenyl) -2- (2-methylphenyl) ethyl) adenosine (DPMA); hexynyladenosine-5' -N-ethylformamide (HE-NECA); 2- [ (2-aminoethyl-aminocarbonylethyl) phenethylamino]-5' -N-ethyl-carboxamidoadenosine (APEC); 2-chloro-N⁶- (3-iodobenzyl) -5' -N-methylcarboxyamidoadenosine (2-Cl-IB-MECA); 2-phenylaminoadenosine (CV 1808); 3 '-aminoadenosine-5' -uronamide; CV therapeutics^TMA small molecule drug Tecadinoson (CVT-510); regadenoson (CVT 3146); and Carisa (CVT 3033); and Aderis Pharmaceuticals^TMSmall molecule drug 2- [2- (4-chlorphenyl) ethoxy]Adenosine (MRE 0094), 1-deoxy-1- [6- [ [ (iodophenyl) methyl]Amino group]-9H-purin-9-yl]-N-methyl- (-D-ribofuranosyluronic acid amide) (CF101), Selodenoson (DTI-0009) and Binodenoson (MRE-0470). Other adenosine receptor agonists are those described or claimed in the following documents: gao et al, JPET, 298: 209-218 (2001); U.S. patent nos. 5,278,150, 5,877,180, 6,232,297; U.S. patent application publication No. 2005/0261236 and PCT publication No. WO/9808855, which are incorporated herein by reference.

Adenosine transport inhibitors

Adenosine transport inhibitors useful in the methods, compositions, and kits of the present invention include 3- [1- (6, 7-diethoxy-2-morpholinoquinazolin-4-yl) piperidin-4-yl ] -piperidine]-1, 6-dimethyl-2, 4(1H, 3H) -quinazolinedione hydrochloride (KF 24345); 6- (4-nitrobenzyl) -thioinosine (NBI) and 6- (2-hydroxy-5-nitrobenzyl) -thioguanosine (NBG); 6- [4- (1-cyclohexyl-1H-tetrazol-5-yl) butoxy]-3, 4-dihydro-2 (1H) -quinolinone (cilostazol); (2-amino-4, 5-dimethyl-3-thienyl) - [3- (trifluoromethyl) phenyl]Methanone (PD 81723); 3, 7-dihydro-3-methyl-1- (5-oxohexyl) -7-propyl-1H-purine-2, 6-dione (propentofylline); 6- [ (4-nitrobenzyl) thio]-9-beta-D-ribofuranosylpurine (nitrobenzyl)Basal thioinosine) (NBMR); 3,4, 5-trimethoxy- (tetrahydro-1H-1, 4-diaza)-1, 4(5H) -diyl) di-3, 1-propanediyl benzoate (delazipril); clandecamine; dipyridamole; and adenosine transport inhibitors described in: fredholm, j.neurochem.62: 563-573(1994), Noji et al, J.Pharmacol.Exp.Ther.300: 200-205 (2002); and Crawley et al; neurosci lett.36: 169-174(1983), each of which is incorporated herein by reference.

Adenosine kinase inhibitors

Adenosine kinase inhibitors may be used as up-regulators of adenosine activity in the methods, compositions, and kits of the invention. Adenosine kinase inhibitors are generally described as nucleoside-like or non-nucleoside-like.

Nucleoside-like adenosine kinase inhibitors

Nucleoside-like adenosine kinase inhibitors useful in the methods, compositions and kits of the invention include 5-iodotubercidin (5IT) and 2-diaryl tubercidin analogs; 5 ' -deoxo-5 ' -deoxy-5-iodotubercidin (5'd-5 IT); and 5 '-deoxo-5' -aminoadenosine (NH)₂dADO). Other nucleoside-like adenosine kinase inhibitors are described in the following references: McGaraughty et al, Current Topics in Medicinal Chemistry 5: 43-58 (2005); ugarkar, j.med.chem.43: 2883 2893 (2000); ugarkar et al, j. med. chem.43: 2894 2905 (2000); kaplan and Coyle, eur.j.pharmacol.1: 1-8 (1998); and Sinclair et al, br.j. pharmacol.5: 1037-1044(2001), each of which is incorporated herein by reference.

Non-nucleoside-like adenosine kinase inhibitors

Non-nucleoside-like adenosine kinase inhibitors useful in the methods, compositions, and kits of the invention include 5-bromopyrrolopyrrolidine; 4-amino-5- (3-bromophenyl) -7- (6-morpholino-pyridin-3-yl) pyrido [2, 3-d ] pyrimidine (ABT-702). Other non-nucleoside-like AK inhibitors are described in the following references: McGaraughty et al, Current Topics in Medicinal Chemistry 5: 43-58 (2005); gomtsyan and Lee, Current Pharmaceutical Design 10: 1093-; jarvis et al, J.pharm.Exp.Ther.295: 1156-1164 (2000); kowaluk et al, J.pharm.Exp.Ther.295: 1165-1174 (2000); and german patent application DE 10141212a1, each of which is incorporated herein by reference.

Phosphodiesterase inhibitors

Several isoenzymes of phosphodiesterase act as regulatory switches by catalyzing the degradation of cAMP to adenosine-5-monophosphate (5' -AMP). Inhibitors of phosphodiesterase can cause an increase in cAMP levels, which in turn can cause an increase in anti-inflammatory effects.

Phosphodiesterase type I inhibitors

Type I PDE inhibitors include (3-alpha, 16-alpha) -etaneremetine-14-carboxylic acid ethyl ester (vinpocetine); 18-methoxymethyl-3-isobutyl-1-methylxanthine (MIMX); 1-carboxy-2, 3,4, 4a, 4b, 5,6, 6a, 6b, 7,8, 8a, 8b, 9, 10, 10a, 14, 16, 17, 17a, 17b, 18, 19, 19a, 19b, 20, 21, 21a, 21b, 22, 23, 23 a-triacontahydro-14-hydroxy-8 a, 10 a-bis (hydroxymethyl) -14- (3-methoxy-3-oxopropyl) -1, 4, 4a, 6, 6a, 17b, 19b, 21 b-octamethyl β -D-glucopyranosiduronic acid (Ks-505 a); cis-5, 6a, 7,8, 9,9 a-hexahydro-2- (4- (trifluoromethyl) benzyl) -5-methyl-cyclopenta (4, 5) imidazo (2, 1-b) purin-4 (3H) -one (SCH 51866); and 2-o-propoxyphenyl-8-azapurin-6-one (zaprinast). Other type I PDE inhibitors are described in U.S. patent application publication nos. 2004/0259792 and 2005/0075795.

Phosphodiesterase type II inhibitors

Type II PDE inhibitors include erythro-9- (2-hydroxy-3-nonyl) adenine (EHNA); 2,3, 6, 7-tetrahydro-9, 10-dimethoxy-3-methyl-2- ((2, 4, 6-trimethylphenyl) imino) -4H-pyrimido (6, 1-a) isoquinolin-4-one (ququicin); ND7001(Neuro3 DPharmaceuticals); and BAY 60-7550(Alexis Biochemicals). Other type II PDE inhibitors are described in U.S. patent application publication No. 2003/0176316.

Phosphodiesterase type III inhibitors

Type III PDE inhibitors include 3-isobutyl-1-methylxanthine (IBMX); 6-dihydro-2-methyl-6-oxo-3, 4' -bipyridine) -5-carbonitrile (miridone) and N-cyclohexyl-4- ((1, 2-dihydro-2-oxo-6-quinolinyl) oxy) -N-methyl-butyramide (cyclohexquinamide). Other type III PDE inhibitors are described in the following patents and patent applications:

EP 0653426, EP 0294647, EP 0357788, EP0220044, EP 0326307, EP 0207500, EP 0406958, EP 0150937, EP 0075463, EP 0272914, and EP0112987, u.s.pat.nos.4,963,561; 5,141,931, 6,897,229, and 6,156,753;

U.S. patent application publication no

2003/0158133, 2004/0097593, 2006/0030611, and 2006/0025463; WO 96/15117; DE 2825048; DE 2727481; DE 2847621; DE 3044568; DE 2837161; and DE 3021792.

Phosphodiesterase type IV inhibitor

Type IV PDE inhibitors include 4- (3-cyclopentyloxy-4-methoxyphenyl) -2-pyrrolidone (rolipram) and 4- (3-butoxy-4-methoxybenzyl) -2-imidazolidinone (Ro 20-1724). Other PDE type IV inhibitors are described in the following patents, patent applications and references:

U.S. Pat. No. 5

3,892,777, 4,193,926, 4,655,074, 4,965,271, 5,096,906, 5,124,455, 5,272,153, 6,569,890, 6,953,853, 6,933,296, 6,919,353, 6,953,810, 6,949,573, 6,909,002, and 6,740,655;

U.S. patent application publication no

2003/0187052，2003/0187257，2003/0144300，2003/0130254，2003/0186974，2003/0220352，2003/0134876，2004/0048903，2004/0023945，2004/0044036，2004/0106641，2004/0097593，2004/0242643，2004/0192701，2004/0224971，2004/0220183，2004/0180900，2004/0171798，2004/0167199，2004/0146561，2004/0152754，2004/0229918，2005/0192336，2005/0267196，2005/0049258，2006/0014782，2006/0004003，2006/0019932，2005/0267196，2005/0222207，2005/0222207，2006/0009481；

PCT publication nos. WO 92/079778 and Molnar-Kimber, k.l. et al, j.immunol, 150: 295A (1993).

Phosphodiesterase type V inhibitors

PDE type V inhibitors are described in the following documents: U.S. patent nos. 6,992,192, 6,984,641, 6,960,587, 6,943,166, 6,878,711 and 6,869,950, and U.S. patent application publication nos

2003/0144296, 2003/0171384, 2004/0029891, 2004/0038996, 2004/0186046, 2004/0259792, 2004/0087561, 2005/0054660, 2005/0042177, 2005/0245544, and 2006/0009481.

Phosphodiesterase type VI inhibitor

Type VI PDE inhibitors include those described in U.S. patent application publication nos. 2004/0259792, 2004/0248957, 2004/0242673, and 2004/0259880.

Phosphodiesterase type VII inhibitors

PDE inhibitors of type VII include those described in the following documents: U.S. patent nos. 6,838,559, 6,753,340, 6,617,357 and 6,852,720; U.S. patent application publication nos. 2003/0186988, 2003/0162802, 2003/0191167, 2004/0214843, and 2006/0009481; PCT publications WO 00/68230; and Martinez et al, j.med.chem.43: 683-689, 2000.

Tricyclic compounds

Methods, sets, useful in the inventionTricyclic compounds in compositions and kits include amitriptyline, amoxapine, clomipramine, desipramine, doxypine, doxepin, imipramine, lofepramine, maprotiline, mianserin, mirtazapine, nortriptyline, octriptyline, hydroxypropiophilin, protriptyline, trimipramine, 10- (4-methylpiperazin-1-yl) pyrido (4, 3-b) (1, 4) benzothiazapine(ii) a 11- (4-methyl-1-piperazinyl) -5H-dibenzo (b, e) (1, 4) diazepine(ii) a 5, 10-dihydro-7-chloro-10- (2- (morpholino) ethyl) -11H-dibenzo (b, e) (1, 4) diazepine-11-ketone; 2- (2- (7-hydroxy-4-dibenzo (b, f) (1, 4) thiazepine-11-yl-1-piperazinyl) ethoxy) ethanol; 2-chloro-11- (4-methyl-1-piperazinyl) -5H-dibenzo (b, e) (1, 4) diazepine(ii) a 4- (11H-dibenzo (b, e) azepine-6-yl) piperazine; 8-chloro-11- (4-methyl-1-piperazinyl) -5H-dibenzo (b, e) (1, 4) diazepine-2-alcohols; 8-chloro-11- (4-methyl-1-piperazinyl) -5H-dibenzo (b, e) (1, 4) diazepineA monohydrochloride salt; (Z) -2-Butenedioate 5H-dibenzo (b, e) (1, 4) diazepine(ii) a Aldizone; amiheptanoic acid; amitriptyline oxide; butiline; imidaclothiz; performing chlorination and leveling; dimetyline; 11- (4-methyl-1-piperazinyl) -dibenzo (b, f) (1, 4) oxazazepine(ii) a 11- (4-methyl-1-piperazinyl) -2-nitro-dibenzo (b, f) (1, 4) oxazazepine(ii) a 2-chloro-11- (4-methyl-1-piperazinyl) -dibenzo (b, f) (1, 4) oxaazepineA monohydrochloride salt; benzhydrazepine; 11- (4-methyl-1-piperazinyl) -dibenzo (b, f) (1, 4) thiazepine(ii) a (ii) dipropionazine; trifluoropropylamine; flupirtine; imipramine N-oxide; aminopropylindole; lofepramine; (ii) melixanthene; metapamine; methipin; methyltriazenfluoranthene; mianserin; mirtazapine; 8-chloro-6- (4-methyl-1-piperazinyl) -morphidine; n-acetyl amoxapine; benzpyrole; norclomipramine; norclozapine; norcetilin; opipramol; oxaprotiline; piperapine; a benzothiidine; third generation of N-propylpyrazine(ii) a Quetiapine; quinuclidine nitrogen(ii) a Tianeptine; tomoxetine; trifluorothianol; clopenthixol; perhexiline; chlorprothixene and thiothixene. Other tricyclic compounds are described in the following patents: for example, U.S. patent No. 4

2,554,736, respectively; 3,046,283, respectively; 3,310,553; 3,177,209, respectively; 3,205,264; 3,244,748, respectively; 3,271,451, respectively; 3,272,826, respectively; 3,282,942, respectively; 3,299,139, respectively; 3,312,689, respectively; 3,389,139, respectively; 3,399,201, respectively; 3,409,640, respectively; 3,419,547, respectively; 3,438,981, respectively; 3,454,554, respectively; 3,467,650, respectively; 3,505,321, respectively; 3,527,766, respectively; 3,534,041, respectively; 3,539,573; 3,574,852, respectively; 3,622,565, respectively; 3,637,660, respectively; 3,663,696, respectively; 3,758,528, respectively; 3,922,305, respectively; 3,963,778, respectively; 3,978,121, respectively; 3,981,917, respectively; 4,017,542, respectively; 4,017,621, respectively; 4,020,096, respectively; 4,045,560, respectively; 4,045,580, respectively; 4,048,223, respectively; 4,062,848; 4,088,647, respectively; 4,128,641, respectively; 4,148,919, respectively; 4,153,629, respectively; 4,224,321, respectively; 4,224,344, respectively; 4,250,094, respectively; 4,284,559, respectively; 4,333,935, respectively; 4,358,620, respectively; 4,548,933, respectively; 4,691,040, respectively; 4,879,288; 5,238,959, respectively; 5,266,570, respectively; 5,399,568, respectively; 5,464,840, respectively; 5,455,246, respectively; 5,512,575, respectively; 5,550,136, respectively; 5,574,173, respectively; 5,681,840, respectively; 5,688,805, respectively; 5,916,889, respectively; 6,545,057, respectively; and 6,600,065 of the group consisting of,

and phenothiazine compounds corresponding to formula (I) of U.S. patent application No. 10/617,424 or 60/504,310.

The standard recommended dosages for several tricyclic compounds are provided in table 6 below. Other standard dosages are provided, for example, in the merck's handbook (17 th edition, MH Beers et al, merck) and in the Physicians' Desk Reference2003 (57 th edition, hygienical chemist et al, hygienical chemist, 2002).

TABLE 6

Compound (I)	Standard dose
		Amoxicillin	200-300 mg/day
Nortriptyline	75-150 mg/day
		Desipramine	100-200 mg/day

Selective 5-hydroxytryptamine reuptake inhibitors

SSRIs can be used in the methods, compositions, and kits of the invention. Suitable SSRIs are cerclamine (e.g., cerclamine hydrochloride); citalopram (e.g. citalopram hydrobromide); flufenamid; cyanodothiepin; dapoxetine; escitalopram (escitalopram oxalate); benfuryloxybenzene (e.g., benfuryloxybenzene hydrochloride); fluoxetine (e.g., fluoxetine hydrochloride); fluvoxamine (e.g., fluvoxamine maleate); ifoxetine; indants (e.g., indants hydrochloride); indenoxazines (e.g., indenoxazine hydrochloride); ritoxetine; milnacipran (e.g., milnacipran hydrochloride); paroxetine (e.g., paroxetine hydrochloride hemihydrate; paroxetine maleate; paroxetine methanesulfonate); sertraline (e.g., sertraline hydrochloride); tametrine hydrochloride; a vitamin A derivative; and zimelidine (e.g., zimelidine hydrochloride).

Structural analogs of cilcamine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein R₁Is C₁-C₄Alkyl radical, R₂Is H or C_1-4Alkyl radical, R₃Is H, C_1-4Alkyl radical, C_2-4Alkenyl, phenylalkyl or cycloalkylalkyl, alkanoyl, phenylalkanoyl having 3 to 6 ring carbon atoms or cycloalkylcarbonyl having 3 to 6 ring carbon atoms, or R₂And R₃Together with the nitrogen atom to which they are attached form a heterocyclic ring saturated with 5 to 7 chain bonds (chainlinks) and which may have, as a second heteroatom not directly attached to the nitrogen atomWith oxygen, sulfur or nitrogen, the latter nitrogen hetero atom possibly carrying C_2-4An alkyl group.

Exemplary structural analogs of cilantrine are 2-methyl-2-amino-3- (3, 4-dichlorophenyl) -propanol, 2-pentyl-2-amino-3- (3, 4-dichlorophenyl) -propanol, 2-methyl-2-methylamino-3- (3, 4-dichlorophenyl) -propanol, 2-methyl-2-dimethylamino-3- (3, 4-dichlorophenyl) -propanol, and any pharmaceutically acceptable salts thereof.

Structural analogues of citalopram are those having the following formula:

and pharmaceutically acceptable salts thereof, wherein R₁And R₂Each independently selected from the group consisting of bromo, chloro, fluoro, trifluoromethyl, cyano and R-CO-, wherein R is C_1-4An alkyl group.

An exemplary citalopram structural analogue (which is thus an SSRI structural analogue according to the present invention) is 1- (4' -fluorophenyl) -1- (3-dimethylaminopropyl) -5-bromo-1, 3-dihydroisobenzofuran (phthalane); 1- (4' -chlorophenyl) -1- (3-dimethylaminopropyl) -5-chloro-1, 3-dihydroisobenzofuran; 1- (4' -bromophenyl) -1- (3-dimethylaminopropyl) -5-chloro-1, 3-dihydroisobenzofuran; 1- (4' -fluorophenyl) -1- (3-dimethylaminopropyl) -5-chloro-1, 3-dihydroisobenzofuran; 1- (4' -chlorophenyl) -1- (3-dimethylaminopropyl) -5-trifluoromethyl-1, 3-dihydroisobenzofuran; 1- (4' -bromophenyl) -1- (3-dimethylaminopropyl) -5-trifluoromethyl-1, 3-dihydroisobenzofuran; 1- (4' -fluorophenyl) -1- (3-dimethylaminopropyl) -5-trifluoromethyl-1, 3-dihydroisobenzofuran; 1- (4' -fluorophenyl) -1- (3-dimethylaminopropyl) -5-fluoro-1, 3-dihydroisobenzofuran; 1- (4' -chlorophenyl) -1- (3-dimethylaminopropyl) -5-fluoro-1, 3-dihydroisobenzofuran; 1- (4' -chlorophenyl) -1- (3-dimethylaminopropyl) -5-1, 3-dihydroisobenzofurancarbonitrile; 1- (4' -fluorophenyl) -1- (3-dimethylaminopropyl) -5-1, 3-dihydroisobenzofurancarbonitrile; 1- (4' -cyanophenyl) -1- (3-dimethylaminopropyl) -5-1, 3-dihydroisobenzofurancarbonitrile; 1- (4' -cyanophenyl) -1- (3-dimethylaminopropyl) -5-chloro-1, 3-dihydroisobenzofuran; 1- (4' -cyanophenyl) -1- (3-dimethylaminopropyl) -5-trifluoromethyl-1, 3-dihydroisobenzofuran; 1- (4' -fluorophenyl) -1- (3-dimethylaminopropyl) -5-1, 3-dihydroisobenzofurancarbonitrile; 1- (4' -chlorophenyl) -1- (3-dimethylaminopropyl) -5-violanyl (ionyl) -1, 3-dihydroisobenzofuran; 1- (4- (chlorophenyl) -1- (3-dimethylaminopropyl) -5-propionyl-1, 3-dihydroisobenzofuran and any pharmaceutically acceptable salts thereof.

Structural analogues of flufenacet are those having the formula:

and pharmaceutically acceptable salts thereof, wherein Hal is chloro, bromo or fluoro, and R is cyano, methoxy, ethoxy, methoxymethyl, ethoxymethyl, methoxyethoxy or cyanomethyl.

An exemplary structural analog of flufenacet is 4' -chloro-5-ethoxycyclopentanone (valrophenone) O- (2-aminoethyl) oxime; 4' -chloro-5- (2-methoxyethoxy) cyclopentanone O- (2-aminoethyl) oxime; 4' -chloro-6-methoxyphenylhexanone O- (2-aminoethyl) oxime; 4' -chloro-6-ethoxyhexanophenone O- (2-aminoethyl) oxime; 4' -bromo-5- (2-methoxyethoxy) cyclopentanone O- (2-aminoethyl) oxime; 4' -bromo-5-methoxyphenylpentanone O- (2-aminoethyl) oxime; 4' -chloro-6-cyanophenone O- (2-aminoethyl) oxime; 4' -chloro-5-cyanophenylpentanone O- (2-aminoethyl) oxime; 4' -bromo-5-cyanophenylpentanone O- (2-aminoethyl) oxime; and any pharmaceutically acceptable salts thereof.

Structural analogs of methylphenidate are those having the formula:

wherein R is₁Represents C_1-4Alkyl or C_2-4Alkynyl or optionally substituted by C_1-4Alkyl radical, C_1-4Alkylthio radical, C_1-4Alkoxy, bromo, chloro, fluoro, nitro, amido, methylsulfonyl, methylenedioxy-substituted phenyl or tetrahydronaphthyl, R₂Represents C_1-4Alkyl or C_2-4Alkynyl radical, R₃Represents hydrogen, C_1-4Alkyl radical, C_1-4Alkoxy, trifluoroalkyl, hydroxy, bromo, chloro, fluoro, methylthio or aralkoxy.

Exemplary structural analogs of methylphenidate are disclosed in examples 7-67 of U.S. Pat. No. 3,912,743, which is incorporated herein by reference.

Structural analogs of fluoxetine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein each R₁Independently is hydrogen or methyl; r is naphthyl or

Wherein R is₂And R₃Each independently is bromine, chlorine, fluorine, trifluoromethyl, C_1-4Alkyl radical, C_1-3Alkoxy or C_3-4An alkenyl group; n and m are each independently 0,1 or 2. When R is naphthyl, it can be alpha-naphthyl or beta-naphthyl.

Exemplary structural analogs of fluoxetine are 3- (p-isopropoxyphenoxy) -3-phenylpropylamine mesylate, N-dimethyl-3- (3 ', 4 ' -dimethoxyphenoxy) -3-phenylpropylamine p-hydroxybenzoate, N-dimethyl-3- (. alpha. -naphthyloxy) -3-phenylpropylamine bromide, N-dimethyl-3- (. beta. -naphthyloxy) -3-phenyl-1-methylpropylamine iodide, 3- (2 ' -methyl-4 ', 5 ' -dichlorophenoxy) -3-phenylpropylamine nitrate, 3- (p-tert-butylphenoxy) -3-phenylpropylamine glutarate, N-tert-butylphenoxy-3-phenylpropylamine, N-hydroxy-N-methyl-3- (p-naphthyloxy) -3-methyl-N-propylamine, N-methyl-3- (p-naphthyloxy) -, Lactic acid N-methyl-3- (2 ' -chloro-p-tolyloxy) -3-phenyl-1-methylpropylamine, citric acid 3- (2 ', 4 ' -dichlorophenoxy) -3-phenyl-2-methylpropylamine, maleic acid N, N-dimethyl-3- (m-anisyloxy) -3-phenyl-1-methylpropylamine, sulfuric acid N-methyl-3- (p-tolyloxy) -3-phenylpropylamine, 2, 4-dinitrobenzoic acid N, N-dimethyl-3- (2 ', 4 ' -difluorophenoxy) -3-phenylpropylamine, dihydrogenphosphoric acid 3- (o-ethylphenoxy) -3-phenylpropylamine, dihydrogenphosphoric acid, a salt of a compound of formula (I), a salt of a compound of formula (II), and a salt of a compound of, Maleic acid N-methyl-3- (2 '-chloro-4' -isopropylphenoxy) -3-phenyl-2-methylpropylamine, succinic acid N, N-dimethyl-3- (2 '-alkyl-4' -fluorophenoxy) -3-phenyl-propylamine, phenylacetic acid N, N-dimethyl-3- (o-isopropoxyphenoxy) -3-phenyl-propylamine, beta-phenylpropionic acid N, n-dimethyl-3- (o-bromophenyloxy) -3-phenyl-propylamine, propionic acid N-methyl-3- (p-iodophenoxy) -3-phenyl-propylamine and decanoic acid N-methyl-3- (3-N-propylphenoxy) -3-phenyl-propylamine.

Structural analogs of fluvoxamine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein R is cyano, cyanomethyl, methoxymethyl, or ethoxymethyl.

Structural analogs of indacene are those compounds having the formula:

or a pharmaceutically acceptable salt thereof, wherein R₁Is a hydrogen atom, C₁-C₄Alkyl or aralkyl in which the alkyl has 1 or 2 carbon atoms, R₂Is hydrogen, C_1-4Alkyl radical, C_1-4Alkoxy or C_1-4Alkylthio, chloro, bromo, fluoro, trifluoromethyl, nitro, hydroxy or amino, the latter optionally being substituted by one or two C_1-4Alkyl, acyl or C_1-4Alkylsulfonyl substitution; a represents-CO or-CH₂-a group; n is 0,1 or 2.

An exemplary indacene structural analog is indolyl-3- (piperidinyl-4-methyl) one; (methoxy-5-indolyl-3) (piperidinyl-4-methyl) one; (chloro-5-indolyl-3) (piperidinyl-4-methyl) one; (indolyl-3) -1- (piperidinyl-4) -3-propanone, indolyl-3-piperidinyl-4-one; (methyl-1-indolyl-3) (piperidinyl-4-methyl) one, (benzyl-1-indolyl-3) (piperidinyl-4-methyl) one; [ (methoxy-5-indolyl-3) -2-ethyl ] -piperidine, [ (methyl-1-indolyl-3) -2-ethyl ] -4-piperidine; [ (indolyl-3) -2-ethyl ] -4-piperidine; (indolyl-3-methyl) -4-piperidine, [ (chloro-5-indolyl-3) -2-ethyl ] -4-piperidine; [ (indolyl-b 3) -3-propyl ] -4-piperidine; [ (benzyl-1-indolyl-3) -2-ethyl ] -4-piperidine; and any pharmaceutically acceptable salts thereof.

Structural analogs of indenoxazine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein R₁And R₃Each represents hydrogen, C_1-4Alkyl or phenyl; r₂Represents hydrogen, C_1-4Alkyl radical, C_4-7Cycloalkyl, phenyl or benzyl; one of the dotted lines refers to a single bond and the others to a double bond, or a tautomeric mixture thereof.

An exemplary indenoxazine structural analog is 2- (7-indenyloxymethyl) -4-isopropylmorpholine; 4-butyl-2- (7-indenyloxymethyl) morpholine; 2- (7-indenyloxymethyl) -4-methylmorpholine; 4-ethyl-2- (7-indenyloxymethyl) morpholine, 2- (7-indenyloxymethyl) -morpholine; 2- (7-indenyloxymethyl) -4-propylmorpholine; 4-cyclohexyl-2- (7-indenyloxymethyl) morpholine; 4-benzyl-2- (7-indenyloxymethyl) -morpholine; 2- (7-indenyloxymethyl) -4-phenylmorpholine; 2- (4-indenyloxymethyl) morpholine; 2- (3-methyl-7-indenyloxymethyl) -morpholine; 4-isopropyl-2- (3-methyl-7-indenyloxymethyl) morpholine; 4-isopropyl-2- (3-methyl-4-indenyloxymethyl) morpholine; 4-isopropyl-2- (3-methyl-5-indenyloxymethyl) morpholine; 4-isopropyl-2- (1-methyl-3-phenyl-6-indenyloxymethyl) morpholine; 2- (5-indenyloxymethyl) -4-isopropyl-morpholine, 2- (6-indenyloxymethyl) -4-isopropyl-morpholine; and 4-isopropyl-2- (3-phenyl-6-indenyloxymethyl) morpholine; and any pharmaceutically acceptable salts thereof.

Structural analogs of milnacipran are those having the formula:

and pharmaceutically acceptable salts thereof, wherein each R independently represents hydrogen, bromine, chlorine, fluorine, C_1-4Alkyl radical, C_1-4Alkoxy, hydroxy, nitro or amino; r₁And R₂Each independently represents hydrogen, C_1-4Alkyl radical, C_6-12Aryl or C_7-14Alkylaryl, optionally substituted by bromo, chloro or fluoro, preferably in the para position, or R₁And R₂Together with the adjacent nitrogen atom form a heterocyclic ring having 5 or 6 members; r₃And R₄Represents hydrogen or C_1-4Alkyl, or R₃And R₄Form a heterocyclic ring with 5 or 6 members with the adjacent nitrogen atom, the heterocyclic ring optionally containing a further heteroatom selected from nitrogen, sulphur and oxygen.

Exemplary structural analogs of milnacipran are 1-phenyl-1-aminocarbonyl-2-dimethylaminomethylcyclopropane; 1-phenyl-1-dimethylaminocarbonyl-2-dimethylaminomethylcyclopropane; 1-phenyl-1-ethylaminocarbonyl-2-dimethylaminomethylcyclopropane; 1-phenyl-1-diethylaminocarbonyl-2-aminomethylcyclopropane; 1-phenyl-2-dimethylaminomethyl-N- (4' -chlorophenyl) cyclopropanecarboxamide; 1-phenyl-2-dimethylaminomethyl-N- (4' -chlorobenzyl) cyclopropanecarboxamide; 1-phenyl-2-dimethylaminomethyl-N- (2-phenylethyl) cyclopropanecarboxamide; (3, 4-dichloro-1-phenyl) -2-dimethylaminomethyl-N, N-dimethylcyclopropanecarboxamide; 1-phenyl-1-pyrrolidin-1-yl-carbonyl-2-morpholinomethylcyclopropane; 1-p-chlorophenyl-1-aminocarbonyl-2-aminomethylcyclopropane; 1-o-chlorophenyl-1-aminocarbonyl-2-dimethylaminomethylcyclopropane; 1-p-hydroxyphenyl-1-aminocarbonyl-2-dimethylaminomethylcyclopropane; 1-p-nitrophenyl-1-dimethylaminocarbonyl-2-dimethylaminomethylcyclopropane; 1-p-aminophenyl-1-dimethylaminocarbonyl-2-dimethylaminomethylcyclopropane; 1-p-tolyl-1-methylaminocarbonyl-2-dimethylaminomethylcyclopropane; 1-p-methoxyphenyl-1-aminomethylcarbonyl-2-aminomethylcyclopropane; and any pharmaceutically acceptable salts thereof.

Structural analogs of paroxetine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein R₁Represents hydrogen or C_1-4Alkyl, the fluorine atom may be in any available position.

Structural analogs of sertraline are those compounds having the formula:

wherein R is₁Selected from hydrogenAnd C_1-4An alkyl group; r₂Is C_1-4An alkyl group; x and Y are independently selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl, C_1-3Alkoxy and cyano; w is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, trifluoromethyl and C_1-3An alkoxy group. Preferred sertraline analogs are in the cis-isomeric configuration. The term "cis-isomerism" refers to NR on the cyclohexene ring₁R₂And the relative orientation of the phenyl moieties (i.e., they are all oriented on the same side of the ring). Because both the 1-carbon and the 4-carbon are asymmetrically substituted, each cis compound has two optically active enantiomeric forms, representing (with respect to the 1-carbon) the cis- (1R) and cis- (1S) enantiomers.

Particularly useful are the following compounds of (1S) -enantiomer or (1S) (1R) racemate, and pharmaceutically acceptable salts thereof: cis-N-methyl-4- (3, 4-dichlorophenyl) -1, 2,3, 4-tetrahydro-1-naphthalenamine; cis-N-methyl-4- (4-bromophenyl) -1, 2,3, 4-tetrahydro-1-naphthylamine; cis-N-methyl-4- (4-chlorophenyl) -1, 2,3, 4-tetrahydro-1-naphthylamine; cis-N-methyl-4- (3-trifluoromethyl-phenyl) -1, 2,3, 4-tetrahydro-1-naphthylamine; cis-N-methyl-4- (3-trifluoromethyl-4-chlorophenyl) -1, 2,3, 4-tetrahydro-1-naphthylamine; cis-N, N-dimethyl-4- (4-chlorophenyl) -1, 2,3, 4-tetrahydro-1-naphthylamine; cis-N, N-dimethyl-4- (3-trifluoromethyl-phenyl) -1, 2,3, 4-tetrahydro-1-naphthylamine; and cis-N-methyl-4- (4-chlorophenyl) -7-chloro-1, 2,3, 4-tetrahydro-1-naphthylamine. Also important are the (1R) -enantiomers of cis-N-methyl-4- (3, 4-dichlorophenyl) -1, 2,3, 4-tetrahydro-1-naphthalenamine.

Structural analogs of zimelidine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein the pyridine nucleus is attached to the adjacent carbon atom at the ortho, meta or para position, wherein R₁Selected from hydrogen, chlorine, fluorine and bromine.

Exemplary zimelidine analogs are (e) -and (z) -3- (4 ' -bromophenyl-3- (2 "-pyridyl) -dimethylallylamine, 3- (4 ' -bromophenyl) -3- (3" -pyridyl) -dimethylallylamine, 3- (4 ' -bromophenyl) -3- (4 "-pyridyl) -dimethylallylamine, and any pharmaceutically acceptable salts thereof.

Structural analogs of any of the above SSRIs are considered herein to be SSRI analogs and, therefore, can be used in any of the methods, compositions, and kits of the invention.

Metabolites

Pharmacologically active metabolites of any of the foregoing SSRIs may also be used in the methods, compositions, and kits of the invention. Exemplary metabolites are didemethyl citalopram, desmethyl sertraline and norfluoxetine.

Analogues

Functional analogs of SSRIs can also be used in the methods, compositions, and kits of the invention. Exemplary functional analogs of SSRI are provided below. One class of SSRI analogs includes SNRI (selective 5-hydroxytryptamine norepinephrine reuptake inhibitors) including venlafaxine, duloxetine, and 4- (2-fluorophenyl) -6-methyl-2-piperazin-1-ylthieno [2, 3-d ] pyrimidine.

Structural analogs of venlafaxine are those having the formula:

and pharmaceutically acceptable salts thereof, wherein a is a moiety of formula (la):

wherein the dotted line represents optional unsaturationAnd a bond; r₁Is hydrogen or alkyl; r₂Is C_1-4An alkyl group; r₄Is hydrogen, C_1-4Alkyl, formyl or alkanoyl; r₃Is hydrogen or C_1-4An alkyl group; r₅And R₆Independently of each other hydrogen, hydroxy, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_1-4Alkanoyloxy, cyano, nitro, alkylmercapto, amino, C_1-4Alkylamino, dialkylamino, C_1-4Alkanoylamino, halo, trifluoromethyl or combinations thereof together as methylenedioxy; n is 0,1, 2,3 or 4.

Structural analogs of duloxetine are those compounds described by the structural formula disclosed in U.S. patent No. 4,956,388, which is incorporated herein by reference.

Other SSRI analogs are 4- (2-fluorophenyl) -6-methyl-2-piperazin-1-ylthieno [2, 3-d ] pyrimidine, 1,2, 3, 4-tetrahydro-N-methyl-4-phenyl-1-naphthylamine hydrochloride; 1,2, 3, 4-tetrahydro-N-methyl-4-phenyl- (E) -1-naphthylamine hydrochloride; n, N-dimethyl-1-phenyl-1-1, 3-dihydroisobenzofuranpropylamine hydrochloride; gamma- (4- (trifluoromethyl) phenoxy) -amphetamine hydrochloride; BP 554; CP 53261; o-desmethylvenlafaxine; WY 45,818; WY 45,881; n- (3-fluoropropyl) paroxetine; lu 19005 and SNRI described in PCT publication No. WO 04/004734.

Standard recommended dose

Table 7 below provides standard recommended dosages for exemplary SSRIs. Other standard dosages are provided in, for example, the Merck's handbook (17 th edition, MH Beers et al, Merck) and the Physicians' Desk Reference2003 (57 th edition, hygienical chemist et al, hygienical chemist, 2002).

TABLE 7

Compound (I)	Standard dose
		Fluoxetine	20-80 mg/day
Sertraline	50-200 mg/day
		Paroxetine	20-50 mg/day
Fluvoxamine	50-300 mg/day
		Citalopram	10-80mg，qid
Escilaria	10mg，qid

Antihistaminic agents

In yet another embodiment of the present invention, the methods, compositions and kits of the present invention employ a histamine receptor antagonist (or analog thereof). Antihistamines are compounds that block the action of histamine. The classes of antihistamines include:

(1) ethanolamines (e.g., diphenhydramine bromide, carbinoxamine, clemastine, dimenhydrinate, diphenhydramine, diphenyllene, and doxylamine);

(2) aminophylline (e.g., pheniramine, pyrilamine, tripelennamine, and triprolidine);

(3) phenothiazines (e.g., diethylazine, prophenamine, meglumine, promethazine, thiethylperazine, and alimemazine);

(4) alkylamines (e.g., acrivastine, brompheniramine, chlorpheniramine, desbrompheniramine, dexchlorpheniramine, fluazinam, and triprolidine);

(5) piperazines (e.g., buclizine, cetirizine, chloroxazine, cyclizine, chlorobenzazine, hydroxyzine);

(6) piperidines (e.g., astemizole, azatadine, cyproheptadine, desloratadine, fexofenadine, loratadine, ketotifen, olopatadine, phenindamine, and terfenadine);

(7) atypical antihistamines (e.g. nitrogen)Statin, levocabastine, methamphetamine, and phentermine).

In the methods, compositions and kits of the invention, antihistamines that are non-sedating and antihistamines that are sedating may be used. Antihistamines particularly desirable for use in the methods, compositions, and kits of the invention are non-sedating antihistamines such as loratadine and desloratadine. Antihistamines having sedative effects can also be used in the methods, compositions, and kits of the invention. Preferred sedating antihistamines for use in the methods, compositions, and kits of the invention are azatadine, diphenhydramine bromide; chlorpheniramine; clemizole; cyproheptadine; dimenhydrinate; diphenhydramine; doxylamine; chlorphenirazine; promethazine; pyrilamine; thiethylperazine and tripelennamine.

Other antihistamines suitable for use in the methods and compositions of the invention are atorvastatin; dimethylaminoacetylphenothiazine; an antazoline; astemizole; nitrogen is present inStatin-like substances(e.g. nitrogen hydrochloride)Statin); a millet product; bepotastine; alcohol acetic theophylline; brompheniramine (e.g., brompheniramine maleate); carbinoxamine (e.g., carbinoxamine maleate); cetirizine (e.g., cetirizine hydrochloride); cetomoxime; a chlorocyazone; clopidogrel; clopidogrel; chlorpheniramine; cinnarizine; clemastine (e.g., clemastine fumarate); chlordiazepam; chlorobenzene tropine; (ii) lornicillin; morpholine (e.g., cyclizine hydrochloride; cyclizine lactate); a tiotropium; dexchlorpheniramine; dexchlorpheniramine maleate; diphenyl forest; stopping for a plurality of times; ebastine; enbramine; emedastine (e.g., emedastine fumarate); epinastine; ibulazine hydrochloride; fexofenadine (e.g., fexofenadine hydrochloride); cisrolidine; hydroxyzines (e.g., hydroxyzine hydrochloride; hydroxynaphthoic acid hydroxyzine); promethazine; isosbestic land; levocabastine (e.g., levocabastine hydrochloride); mebhydrolin; mequitazine; mesafurine; mesalamine; a Metro dragon; mizolastine; olopatadine (e.g., olopatadine hydrochloride); an oxfenadrin; phenindamine (e.g., phenindamine tartrate); (ii) pheniramine; phenytoxan; para-tolylhydramine; fluazinam; (ii) setastine; talalastine; terfenadine; (ii) west nile diamine; thiirammonium (e.g., thiirammonium methidate); sonlazine hydrochloride; topiramine; triprolidine and tritoquinoline.

Structural analogs of antihistamines can also be used according to the invention. Antihistamine analogs include, but are not limited to: 10-piperazinylpropylphenothiazine; 4- (3- (2-chlorophenothiazin-10-yl) propyl) -1-piperazineethanol dihydrochloride; 1- (10- (3- (4-methyl-1-piperazinyl) propyl) -10H-phenothiazin-2-yl) - (9C1) 1-propanone; 3-methoxycyproheptadine; 4- (3- (2-chloro-10H-phenothiazin-10-yl) propyl) piperazine-1-ethanol hydrochloride; 10, 11-dihydro-5- (3- (4-ethoxycarbonyl-4-phenylpiperidino) propylidene) -5H-dibenzo (a, d) cycloheptene; aceiproplane; vinegar perphenazine; alimemazine (e.g., alimemazine hydrochloride); aminopropionazine; benzimidazole; butapiperazine; carphenazine; (ii) chlorpyrifos; clomiprazole; pantoprazole; demethylated astemizole; demethylated cyproheptadine; diethylzines (e.g., diethylzines hydrochloride); prophenamine (e.g., prophenamine hydrochloride); 2- (p-bromophenyl- (p' -tolyl) methoxy) -N, N-dimethyl-ethylamine hydrochloride; n, N-dimethyl-2- (diphenylmethoxy) -ethylamine methyl bromide; EX-10-542A; (ii) fenethylzine; furazol; methyl 10- (3- (4-methyl-1-piperazinyl) propyl) phenothiazin-2-yl ketone; lerisetron; methoxamine; (ii) methodazine; isofenbutazine; n-demethylpromethazine; niperprazole; dethioridazine; perphenazine (e.g., perphenazine enanthate); 10- (3-dimethylaminopropyl) -2-methylsulfanyl-phenothiazine; 4- (dibenzo (b, e) thiepin-6 (11H) -ylidene) -1-methyl-piperidine hydrochloride; prochlorperazine; promethazine; propionyl promethazine (e.g., propionyl promethazine hydrochloride); rotundamine; rupatadine; sch 37370; sch 434; a tecastemizole; thiiramine; perphenazine acetate; thioridazine (e.g., thioridazine hydrochloride) and 3- (10, 11-dihydro-5H-dibenzo (a, d) cyclohepten-5-ylidene) -tropane.

Other compounds suitable for use in the present invention are: AD-0261; AHR-5333; alexanastine; apremidine; ATI-19000; pimavastin; bilastine; bron-12; a caprostatin; chlorpheniramine; lofuradine; a corsym; DF-1105501; DF-11062; DF-1111301; EL-301; (ii) ebazine; F-7946T; f-9505; HE-90481; HE-90512; hivenyl; HSR-609; ecteinascidin; KAA-276; KY-234; laminarin; LAS-36509; LAS-36674; levocetirizine; levoprotiline; metoclopramide; NIP-531; nobiletin; an oxamil; PR-881-884A; (ii) quinisutamazine; a rosuvastatin; seleno bis (phenethyl) thioate; SK & F-94461; SODAS-HC; tagolipizine; TAK-427; temastine; UCB-34742; UCB-35440; VUF-K-8707; wy-49051 and ZCR-2060.

Still other compounds suitable for use in the present invention are described in the following patents: U.S. Pat. No. 5

3,956,296, respectively; 4,254,129, respectively; 4,254,130, respectively; 4,282,833, respectively; 4,283,408, respectively; 4,362,736, respectively; 4,394,508, respectively; 4,285,957, respectively; 4,285,958, respectively; 4,440,933, respectively; 4,510,309, respectively; 4,550,116, respectively; 4,692,456, respectively; 4,742,175, respectively; 4,833,138, respectively; 4,908,372, respectively; 5,204,249, respectively; 5,375,693, respectively; 5,578,610, respectively; 5,581,011, respectively; 5,589,487, respectively; 5,663,412, respectively; 5,994,549, respectively; 6,201,124, respectively; and 6,458,958.

Standard recommended dose

Table 8 shows the standard recommended dosages for several exemplary antihistamines. Other standard dosages are provided in, for example, the merck handbook (17 th edition, MH Beers et al, merck corporation.) and the Physicians' Desk Reference2003 (57 th edition, hygienical chemist et al, hygienical chemist, 2002).

TABLE 8

Compound (I)	Standard dose
		Desloratadine	5 mg/once a day
Thielazine	10 mg/day for 1-3 times
		Bromophenylhydramine	12.5-25 mg/1 time every 4-6 hours
Promethazine	25 mg/day for 2 times
		Cyproheptadine	12-16 mg/day
ChlorineRetidine (TAIL)	10 mg/day
		Clilimidazole	100mg given in IV or IM
Azatadine	1-2 mg/day for 2 times

Cetirizine	5-10 mg/day for 1 time
		Chlorpheniramine	2 mg/1 time every 6 hours or 4 mg/1 time every 6 hours
Medicine for curing motion sickness	50-100 mg/1 time every 4-6 hours
		Diphenylamine	25 mg/1 time every 4-6 hours or 38 mg/1 time every 4-6 hours＊
Doxylamine	25 mg/1 time per day or 12.5 mg/1 time every 4 hours＊
		Fexofenadine	60 mg/2 times daily or 180 mg/once daily
Meclozine	25-100 mg/day
		Mepiranamine	30 mg/1 time every 6 hours
Tripirnamine	25-50 mg/1 every 4 to 6 hours or 100 mg/2 times daily (sustained release)＊

Loratadine

Loratadine (CLARITIN) is a tricyclic piperidine that is a selective peripheral histamine H1 receptor antagonist. Loratadine and its structural and functional analogs such as piperidines, tricyclopiperidines, histamine H1-receptor antagonists may be used in the anti-immunoinflammatory combinations of the present invention for the treatment of immunoinflammatory disorders, transplant organ rejection, and graft versus host disease.

Functional and/or structural analogs of loratadine include other H1-receptor antagonists, such as AHR-11325, atorvastatin, antazoline, astemizole, azatadine, nitrogenStatin, bromofinasteride, carpidine, cetirizine, chlorpheniramine, clociclazine, clemastine, cyproheptadine, descarboethoxyloratadine, dexchlorpheniramine, dimenhydrinate, diphenyllene, diphenhydramine, ebastine, fexofenadine, hydroxyzine ketotifen, lodoxamide, levocabastine, meglumine, mequitazine, oxamide, pheniramine, pyrilamine, promethazine, pyrilamine, setastine, tamsulosinBases, temastine, terfenadine, alimemazine, tripelennamine, triprolidine, utrizine and similar compounds (e.g. as described in U.S. patent No.: U.S. Pat No.: Compound No.: U.S. patent No.)

3,956,296, 4,254,129, 4,254,130, 4,283,408, 4,362,736, 4,394,508, 4,285,957, 4,285,958, 4,440,933, 4,510,309, 4,550,116, 4,692,456, 4,742,175, 4,908,372, 5,204,249, 5,375,693, 5,578,610, 5,581,011, 5,589,487, 5,663,412, 5,994,549, 6,201,124, and 6,458,958).

Loratadine, cetirizine, and fexofenadine are second generation H1 receptor antagonists lacking the sedative effects of many first generation H1 receptor antagonists. Piperidine H1 receptor antagonists include loratadine, cyproheptadine hydrochloride (PERIACTIN), and phenindamine tartrate (NOLAHIST). Piperazine H1 receptor antagonists include hydroxyzine hydrochloride (ATARAX), hydroxyzine pamoate (VISTARIL), cyclizine hydrochloride (MAREZINE), cyclizine lactate, and meclizine hydrochloride.

Standard recommended dose

Oral loratadine formulations include tablets, orally disintegrating tablets, and syrups. The loratadine tablets contained 10mg of micronized loratadine. The loratadine syrup contained 1mg/ml of micronized loratadine, and the orally disintegrating tablet (fast disintegrating tablet) contained 10mg of micronized loratadine, which rapidly disintegrated in the oral cavity. Although the recommended dose varies depending on the condition of the patient, the standard recommended dose is provided below. Loratadine is typically administered in a 10mg dose 1 time per day, although other daily doses useful in the anti-immunoinflammatory combination of the present invention include 0.01-0-05mg, 0.05-1mg, 1-3mg, 3-5mg, 5-10mg, 10-15mg, 15-20mg, 20-30mg, and 30-40 mg.

Loratadine is rapidly absorbed upon oral administration. It is metabolized in the liver by cytochrome P4503a4 and cytochrome 4502D6 to descarboethoxyloratadine. Loratadine metabolites may also be used in the anti-immunoinflammatory combination of the present invention.

Butylbenzene hydroxy acid

Butylbenzoic acid (p-butoxyphenylacetylhydroxamic acid, 4-butoxyphenylacetylhydroxamic acid) is used as antipruritic and anti-inflammatory agent.

Butylbenzoic acid analogs include, but are not limited to: 2- (p-propoxyphenyl) acetyl hydroxamic acid; 2- (4-butoxy-m-tolyl) acetylhydroxamic acid; 2- (4-butoxy-3-ethylphenyl) acetylhydroxamic acid; 2- (4-butoxy-3-chlorophenyl) acetyl hydroxamic acid.

Indications for bufexamac include, but are not limited to, eczema, dermatitis, pruritus and hemorrhoids. Bufexamic acid is currently available in ointments, oral preparations, gel preparations, creams or suppositories. Ointments are typically applied 1-3 times per day. The concentration of bufexamac in the ointment, cream, suppository and gel is 0.25-0.5%. In addition, oral doses of bufexamac are commonly administered to rheumatoid arthritis patients. In this case, 250mg of bufexamac was administered 4 times a day. Clinical doses of bufexamac up to 2.0 g/day have been administered.

Calcium channel inhibitors

Calcium channel inhibitors may be used in the methods, compositions, and kits of the invention. Calcium channel inhibitors include verapamil, anipamil, bepridil, gallopamil, delavapamil, falipanim, tiapamil, nifedipine, amlodipine, dalodipine, felodipine, isradipine, lanicalipine, nicardipine, nimodipine, nisoldipine, nitrendipine, ryosidie, diltiazemCinnarizine and flunarizine, BAY-m 4786 and dipine.

Anti-dyskinesia drugs

Anti-dyskinetic compounds useful in the methods, compositions, and kits of the present invention include D-AP5(D (-) -2-amino-5-phosphonovalerate), CGS 19755 (4-phosphonomethyl-2-piperidinecarboxylic acid), CGP37849(D, L- (E) -2-amino-4-methylphosphono-3-pentanoic acid), LY233053 (cis- (+ -) -4- (2H-tetrazol-5-yl) methyl-piperidine-2-carboxylic acid), AIDA (1-aminoindan-1, 5(RS) -dicarboxylic acid), (S) - (+) -CBPG ((S) - (+) -2- (3' -carboxy-bicyclo (1.1.1. -) pentyl) glycine), CPCCOEt (cyclopropane (b) chromene-1 a-carboxylate), EGLU ((s) - (α) -ethylglutamate), LY307452(2s, 4 s-2-amino-4- (4, 4-diphenylbutan-1-yl) pentan-1, 5-dioic acid), LY341495(2 s-2-amino-2- (1s, 2 s-2-carboxy-cycloprop-1-yl) -3- (xanthen-9-yl) propanoic acid), PCCG-4(2s, 1's, 2's, 3 ' R) -2- (2 ' -carboxy-3 ' -phenylcyclopropyl) glycine), 4-CPG (4-carboxyphenylglycine), remacemide, dextromethorphan ((+) -3-hydroxy-N-methyllevorphanol), its metabolites dextrorphan ((+) -3-hydroxy-N-methyllevorphanol), amantadine (1-aminodiamond), memantine (3, 5-dimethylamino diamond (adamantone)), pyrroloquinoline quinone, and cis-4- (phosphonomethyl) -2-piperidinecarboxylic acid.

Anticoccidial compounds

Anticoccidial compounds can be used in the methods, compositions, and kits of the invention. Such compounds include salinomycin, monensin, narasin, nitrabamine, maduramicin, a combination of nitrabamine and narasin, diclazuril, dinitramine, bromoclopidogrel, bischlorobenaminoguanidine, apracloniam and clopidol. Other anticoccidial compounds are described in the following patents: U.S. patent No. 4,582,822; 4,824,863, respectively; 5,552,386, respectively; 6,384,052 and 6,528,531.

Treatment of

The invention features a method of treating an immunoinflammatory disorder. Although the examples describe combinations of 2 drugs, it is understood that combinations of multiple drugs are generally desirable. For example, methotrexate, hydroxychloroquine, and sulfasalazine are commonly administered for the treatment of rheumatoid arthritis, and may also be used in the methods, compositions, and kits of the present invention. Additional treatments are described below.

The methods, compositions, and kits of the present invention are expected to be more effective than other methods, compositions, and kits. By "more effective" is meant that the method, composition, or kit exhibits greater efficacy, less toxicity, is safer, more convenient, better tolerated or less expensive, or provides more satisfactory treatment than another method, composition, or kit of control.

Psoriasis disease

The methods, compositions and kits of the invention can be used to treat psoriasis. If desired, one or more typical anti-psoriasis agents for treating psoriasis may be used in the methods, compositions, and kits of the present invention. Such agents include biologicals (e.g., alfaxacit, infliximab, adalimumab, efavirenz, etanercept, and CDP-870), small molecule immunomodulators (e.g., VX 702, SCIO 469, doramapimod, RO30201195, SCIO 323, DPC333, pralnacasan, mycophenolate, and merimepodib), non-steroidal immunophilin-dependent immunosuppressants (e.g., cyclosporine, tacrolimus, pimecrolimus, ABT-281, and ISAtx247), vitamin D analogs (e.g., calcipotriene, calcipotriol), psoralens (e.g., methoxsalen), retinoid (e.g., acitretin, tazarotene), DMARD (e.g., methotrexate), dithranol, topical glucocorticoids (e.g., clobetasol, triamcinolone, betamethasone, hydrocortisone, clobetasol propionate, diflorasone, halcinolone, fluocinolone acetonide, and systemic corticosteroids (e.g., glucocorticoids, e.g., glucocorticosteroids, glucocorticoids, triamcinolone, and CDP-870) Dexamethasone).

Atopic dermatitis

The methods, compositions, and kits of the invention are useful for treating atopic dermatitis. If desired, one or more atopic dermatitis medications, typically used to treat atopic dermatitis, may be used in the methods, compositions, and kits of the present invention. Such drugs include topical and systemic non-steroidal immunophilin-dependent immunosuppressants (e.g., cyclosporine, tacrolimus, pimecrolimus, ABT-281, and ISAtx247), topical glucocorticosteroids (e.g., clomazone, triamcinolone, betamethasone, hydrocortisone, halobetasol, diflorasone, mometasone, halcinonide, fluticasone), systemic glucocorticosteroids (e.g., prednisone, dexamethasone), and antihistamines (e.g., hydroxyzine, loratadine, cetirizine, diphenhydramine, cyproheptadine, fexofenadine).

Dermatitis of hand

The methods, compositions, and kits of the invention are useful for treating hand dermatitis. If desired, one or more hand dermatitis medications, typically used to treat hand dermatitis, may be used in the methods, compositions, and kits of the present invention. Such drugs include topical and systemic non-steroidal immunophilin-dependent immunosuppressants (e.g., cyclosporine, tacrolimus, pimecrolimus, ABT-281, and ISAtx247), topical glucocorticosteroids (e.g., clomazone, triamcinolone, betamethasone, hydrocortisone, halobetasol, diflorasone, mometasone, halcinonide, fluticasone), systemic glucocorticosteroids (e.g., prednisone, dexamethasone), antihistamines (e.g., hydroxyzine, loratadine, cetirizine, diphenhydramine, cyproheptadine, fexofenadine), and emollients, creams, humectants, and lotions.

Actinic keratosis

Actinic keratosis can be treated with the methods, compositions, and kits of the invention. If desired, one or more hand dermatitis medications, typically used to treat hand dermatitis, may be used in the methods, compositions, and kits of the present invention. Such drugs include chemotherapeutic drugs (e.g., 5-fluorouracil, imiquimod), non-steroidal anti-inflammatory drugs (e.g., diclofenac), topical retinoids (e.g., adapalene), and photodynamic therapy with topical aminolevulinic acid.

Basal cell carcinoma

The methods, compositions, and kits of the invention can be used to treat basal cell carcinoma. If desired, one or more basal cell carcinoma drugs, typically used to treat basal cell carcinoma, may be used in the methods, compositions, and kits of the present invention. Such drugs include chemotherapeutic drugs (e.g., 5-fluorouracil, imiquimod).

Chronic obstructive pulmonary disease

In one embodiment, the methods, compositions and kits of the invention are used to treat Chronic Obstructive Pulmonary Disease (COPD). If desired, one or more drugs typically used to treat COPD may be used in the methods, compositions and kits of the present invention. Such drugs include xanthines (e.g., choline theophyllinate), anticholinergic compounds (e.g., ipratropium, tiotropium), biologicals, small molecule immunomodulators and beta receptor agonists/bronchodilators (e.g., ibuterol sulfate, mefenapyr mesylate, epinephrine, formoterol fumarate, isoproterenol, levalbuterol hydrochloride, metaproterenol sulfate, pirbuterol acetate, salmeterol xinafoate, and terbutaline).

Inflammatory bowel disease

The methods, compositions, and kits of the invention can be used to treat inflammatory bowel disease. If desired, one or more drugs typically used in the treatment of inflammatory bowel disease may be used in the methods, compositions, and kits of the invention. Such agents include biologicals (e.g., infliximab, adelimumab, and CDP-870), small molecule immunomodulators (e.g., VX 702, SCIO 469, doramapimod, RO30201195, SCIO 323, DPC333, pranalcasan, mycophenolate mofetil, and merimepodib), non-steroidal immunophilin-dependent immunosuppressants (e.g., cyclosporine, tacrolimus, pimecrolimus, ABT-281, and ISAtx247), 5-aminosalicylic acids (e.g., mesalamine, sulfasalazine, disodium salazine, and sodium olsalazine), DMARDs (e.g., methotrexate and azathioprine), and alosetron.

Rheumatoid arthritis

The methods, compositions and kits of the invention can be used to treat rheumatoid arthritis. If desired, one or more drugs typically used in the treatment of rheumatoid arthritis may be used in the methods, compositions, and kits of the invention. Such drugs include NSAIDs (e.g., naproxen sodium, diclofenac potassium, aspirin, sulindac, diflunisal, piroxicam, indomethacin, ibuprofen, nabumetone, choline magnesium trisalicylate, sodium salicylate, salicylsalicylic acid (salsalate), fenoprofen, flurbiprofen, ketoprofen, meclofenamate sodium, meloxicam, oxaprozin, sulindac, and tolmetin), COX-2 inhibitors (e.g., rofecoxib, celecoxib, valdecoxib, and clomexib), biologicals (e.g., inflixb, adelimumab, etanercept, CDP-870, rituxib, and atlizb), small molecule immunomodulators (e.g., VX 702, SCIO 469, dorapipimarib, RO 01130295, SCIO 323, cyclosporin 333, pranalan, mycophenolate, and merimeprobamate), immunophilin-dependent immunosuppressants (e.g., VX, cyclosporin, tacrolimus, and merimeprobamate), and immunophilin-dependent immunosuppressants (e, e.g., naproxen, and pharmaceutically acceptable salts thereof), and pharmaceutically acceptable salts thereof, Pimecrolimus, ABT-281, and ISAtx247), 5-aminosalicylic acids (e.g., mesalamine, sulfasalazine, disodium balsalazide, and sodium olsalazine), DMARDs (e.g., methotrexate, leflunomide, minocycline, auranofin, gold sodium thiomalate, gold thioglucose, and azathioprine), hydroxychloroquine sulfate, and penicillamine.

Asthma (asthma)

The methods, compositions and kits of the invention can be used to treat asthma. One or more drugs typically used in the treatment of asthma may be used in the methods, compositions and kits of the present invention, if desired. Such drugs include β 2 agonists/bronchodilators/leukotriene modulators (e.g., zafirlukast, montelukast, and zileuton), biologicals (e.g., omazuril), small molecule immunomodulators, anticholinergic compounds, xanthine, epinephrine, guaifenesin, cromolyn sodium, nedocromil sodium, and potassium iodide.

Osteoarthritis

The methods, compositions and kits of the invention can be used to treat osteoarthritis or the pain, tenderness, mobility impairment, soft tissue bulking or bone bulking associated therewith. If desired, one or more drugs typically used in the treatment of osteoarthritis may be used as a replacement for or in addition to corticosteroids in the methods, compositions, and kits of the invention. Such agents include NSAIDs (e.g., naproxen sodium, diclofenac potassium, aspirin, sulindac, diflunisal, piroxicam, indomethacin, ibuprofen, nabumetone, choline magnesium trisalicylate, sodium salicylate, salicylsalicylic acid (salsalate), fenoprofen, flurbiprofen, ketoprofen, meclofenamate sodium, meloxicam, oxaprozin, sulindac, and tolmetin), COX-2 inhibitors (e.g., rofecoxib, celecoxib, valdecoxib, and clomexib), biologicals (e.g., infliximab, adelimumab, etanercept, CDP-870, rituximab, and atlizumab), small molecule immunomodulators (e.g., VX 702, SCIO 469, doramapi, DMARO 30295, SCIO 323, DPC333, pranallan, mycophenolate, and merzid), methotrexate (e.g., minoxidin, flunomifoil RD, flunomic, flunomimetical D, and trimetrexate, Auranofin, aurothiomalate, aurothioglucose, and azathioprine), xanthines (e.g., theobromine, theophylline, aminophylline, and caffeine), NsIDI (e.g., cyclosporin, tacrolimus, pimecrolimus, and ISAtx247), vitamin D analogs (e.g., calcipotriol, tacalcitol, and maxacalcitol), psoralens (e.g., methoxsalen and trimetaline), retinoids (e.g., tretinoin, isotretinoin, and acetretin), 5-aminosalicylic acid (e.g., mesalamine, sulfasalazine, balsalazide disodium, and olsalazine), hydroxychloroquine sulfate, penicillamine, or analogs thereof.

Eye diseases

The methods, compositions and kits of the invention can be used for the treatment of ocular diseases. Unless adversely affecting the intended purpose of use, the prophylactic and therapeutic agents of the present invention may contain or may be used together with other suitable pharmacologically effective substances, such as dipivefrin (e.g., 0.1% dipivefrin eye drops), anti-VEGF therapy (e.g., pegaptanib (MACUGEN), ranibizumab, anecortave and squalamine lactate), photodynamic therapy (e.g., VIUDYNE (verteporfin)), corticosteroids (e.g., dexamethasone, prednisolone), NSAIDs (e.g., diclofenac sodium, pirprofen), antiallergic agents (e.g., lisabenz, ketotifen fumarate, cromolyn sodium), antihistamines (e.g., diphenhydramine hydrochloride), glaucoma therapeutic agents (e.g., pilocarpine hydrochloride, physostigmine salicylate, timolol, isopropyl unoprostone), artificial tears, antibiotics (e.g., gentamycin sulfate, gentamycin, and, Neomycin sulfate, tobramycin, benzylpenicillin, cefmenoxime, erythromycin ethylsuccinate, colistin, oxytetracycline, polymyxin B, chloramphenicol, micronomicin, dibekacin, sisomicin, sulfamethizole, sulfamethoxazole, ofloxacin, norfloxacin, lomefloxacin hydrochloride, enoxacin, ciprofloxacin hydrochloride, cinoxacin, sparfloxacin, tosufloxacin tosylate, nalidixic acid, pipemidic acid trihydrate, fleroxacin, levofloxacin), antiviral agents (e.g., idodine, aciclovir), and antifungal agents (e.g., pimaricin, fluconazole, miconazole, amphotericin B, flucytosine, itraconazole).

Administration of drugs

In particular embodiments of any of the methods of the invention, the compounds are administered within 14 days of each other, within 10 days of each other, within 5 days of each other, within 24 hours of each other, or simultaneously. The compounds may be formulated together in a single composition, or may be formulated and administered separately. One or both compounds defined herein may be administered in low or high doses. It may be desirable to administer other compounds to the patient, such as corticosteroids, humectants, NSAIDs (e.g., naproxen sodium, diclofenac potassium, aspirin, sulindac, diflunisal, piroxicam, indomethacin, ibuprofen, nabumetone, choline magnesium trisalicylate, sodium salicylate, salicylsalicylic acid, fenoprofen, flurbiprofen, ketoprofen, meclofenamate sodium, meloxicam, oxaprozin, sulindac, and tolmetin), COX-2 inhibitors (e.g., rofecoxib, celecoxib, valdecoxib, and lumiracoxib), glucocorticoid receptor modulators, or DMARDs. The combination therapy of the present invention is particularly useful for the treatment of immunoinflammatory disorders in combination with other drugs that modulate the immune response to positively affect the disorder's biological or small molecules. Such agents include those that exclude critical inflammatory cells, affect cell adhesion, or affect cytokines involved in the immune response. This last class includes drugs that mimic or increase the activity of anti-inflammatory cytokines, such as IL-10, and drugs that inhibit the activity of pro-inflammatory cytokines, such as IL-6, IL-1, IL-2, IL-12, IL-15, or TNF α. Drugs that inhibit TNF α include etanercept, adelimumab, infliximab, and CDP-870. In this example (drug that blocks tnpa effect), the combination therapy reduces cytokine production, and etanercept or infliximab act on a residual fraction of inflammatory cytokines, providing enhanced treatment. Small molecule immunomodulators include, for example, p38MAP kinase inhibitors such as VX 702, SCIO 469, doramapimod, RO30201195, SCIO 323, TACE inhibitors such as DPC333, ICE inhibitors such as pranalcasan, and IMPDH inhibitors such as mycophenolate and merimepodib.

The therapy according to the invention can be performed alone or in combination with another therapy and can be provided at home, in a doctor's office, in a clinic, in a hospital's clinic, or in a hospital. Treatment is optionally initiated at the hospital so that the physician can closely observe the efficacy, make any adjustments required, or can initiate treatment on an outpatient basis. The duration of treatment depends on the type of disease or condition being treated, the age and condition of the patient, the stage and type of progression of the patient's disease, and the patient's response to the treatment. In addition, persons at greater risk of developing inflammatory disease (e.g., persons suffering from age-related hormonal variations) may receive treatment to inhibit or delay the onset of symptoms.

Routes of administration for the various embodiments include, but are not limited to, topical, transdermal, nasal, and systemic administration (e.g., intravenous, intramuscular, subcutaneous, inhalation, rectal, buccal, vaginal, intraperitoneal, intraarticular, ocular, otic, or oral administration). As used herein, "systemic administration" refers to all non-transdermal routes of administration, particularly excluding topical and transdermal routes of administration.

In combination therapy, the dosage and frequency of administration of each component of the combination can be independently controlled. For example, one compound may be administered 3 times per day, while a second compound may be administered 1 time per day. The combination therapy may be administered in an on-and-off cycle that includes a rest period so that the patient's body has an opportunity to recover from hitherto unexpected side effects. The compounds may also be formulated together so that a single administration releases both compounds.

The compounds of the combination may be formulated in a variety of ways known in the art. For example, the first drug and the second drug may be formulated together or separately. It is desirable that the first agent and the second agent be formulated together for simultaneous or near simultaneous administration. Such coformulation compositions may include the two drugs together in the same pill, cream, ointment, foam, capsule, liquid formulation, and the like. It will be appreciated that when referring to the combination preparations of the invention, the formulation techniques employed may also be applied to the preparation of each of the medicaments of the combination as well as to other combinations of the invention (e.g. glucocorticoid receptor modulator combinations in place of corticosteroids). By using different formulation strategies for different drugs, the pharmacokinetic properties of each drug can be suitably matched.

The separately or separately formulated drugs may be packaged together as a kit. Non-limiting examples include kits comprising, for example, 2 pills, pills and powders, suppositories and liquid formulations in vials, two topical ointments, creams, foams, and the like. The kit may include optional components to facilitate administration of unit doses to a patient, such as vials for reconstituting powders, syringes for injections, customized IV delivery systems, inhalers, and the like. In addition, the unit dose kit may contain instructions for the preparation and administration of the composition. The kit can be formulated as a single unit dose for 1 patient, multiple use for a particular patient (at a constant dose or where the activity of each compound may vary as treatment progresses); or the kit may contain a plurality of doses suitable for administration to a plurality of patients ("bulk packaging"). The kit components may be assembled into cartons, blister packs, bottles, test tubes, and the like.

Topical formulations

For the prevention and/or treatment of inflammatory skin diseases it is desirable that the combination of the invention is formulated for topical administration. Topical formulations that can be used with the combination of the invention include, but are not limited to, ointments, foams, lotions, gels, sticks, sprays, solutions (e.g., for soaking when containing bath salts), and creams.

Any conventional pharmacologically and cosmetically acceptable vehicle may be used. The compounds may also be administered, for example, in a liposome formulation that allows the compounds to enter the skin. Such liposome formulations are described in the following patents: U.S. Pat. No. 5

5,169,637；5,000,958；5,049,388；4,975,282；5,194,266；5,023,087；5,688,525；5,874,104；5,409,704；5,552,155；5,356,633；5,032,582；4,994,213；

And PCT publication No. WO 96/40061. Examples of other suitable vehicles are described in U.S. Pat. No. 4,877,805 and EP publication No. 0586106A 1. Suitable vehicles for the present invention may also include mineral oil, petrolatum, polydecene, stearic acid, isopropyl myristate, polyoxyethylene 40 stearate, stearyl alcohol or vegetable oil.

The formulations may include various conventional coloring agents, fragrances, thickeners (e.g., xanthan gum), preservatives, emollients (e.g., hydrocarbon oils, waxes, or silicones), demulcents, solubilizing excipients, dispersants, penetration enhancers, plasticizers, preservatives, stabilizers, demulsifiers, humectants, emulsifiers, moisturizers, astringents, deodorants, and the like, which may be added to provide additional benefits and to improve the feel and/or appearance of the topical formulation.

When a drug is poorly soluble at physiological pH, one or more solubilizing excipients may be an essential ingredient in a topical formulation.

Solubilization refers to the improvement of solubility due to the surface active compounds that can convert water-insoluble or nearly water-insoluble substances into clear or opalescent aqueous solutions without changing the chemical structure of these substances in the process.

The solute formed is noteworthy due to the fact that: in the molecular association, micelles of surface-active compounds formed in aqueous solution, the substances are present in dissolved form. The resulting solution was visually clear to opalescent.

Solubilizing excipients that can be used in the formulations of the present invention include, but are not limited to, compounds belonging to the following classes: polyethoxylated fatty acids, PEG-fatty acid diesters, PEG-fatty acid mono-and diester mixtures, polyethylene glycol glycerol fatty acid esters, alcohol-oil transesterification products, polyglycerolated fatty acids, propylene glycol fatty acid esters, mixtures of propylene glycol esters and glycerides, mono-and diglycerides, sterols and sterol derivatives, polyethylene glycol sorbitan fatty acid esters, polyethylene glycol alkyl ethers, sugar esters, polyethylene glycol alkylphenols, polyoxyethylene-polyoxypropylene block copolymers, sorbitan fatty acid esters, lower alcohol fatty acid esters, ionic surfactants, tocopherol esters, and sterol esters. Each of these classes of excipients is commercially available and known to those skilled in the formulation art.

Ophthalmic preparation

Ophthalmic formulations include, but are not limited to, ophthalmic injections such as intravitreal, tenon's capsule (subcontens), subconjunctival, periocular, retrobulbar injections; topical ophthalmic aqueous solutions such as suspensions, creams and gels; biodegradable and non-biodegradable implants in the eye; an implant inserted through an incision made in the wall of the eye or sutured around the eyeball; tacks for intraocular drug release; and bioadhesive ocular inserts.

For topical ophthalmic administration, the novel formulations of the present invention may take the form of solutions, gels, creams, suspensions or solid inserts formulated so that the unit dose contains a therapeutically effective amount of each active ingredient or some submultiple thereof.

Typical ophthalmically acceptable carriers for the novel formulations are: such as water, mixtures of water and water miscible solvents such as lower alkanols or aralkanols, vegetable oils, polyalkylene glycols, petroleum based gels, ethyl cellulose, ethyl oleate, carboxymethyl cellulose, polyvinylpyrrolidone, isopropyl myristate and other conventionally used acceptable carriers. The pharmaceutical formulations may also contain non-toxic auxiliary substances such as emulsifying agents, preservatives, wetting agents, base agents and the like, for example polyethylene glycols 200, 300, 400 and 600, polyethylene glycols 1,000, 1,500, 4,000, 6,000 and 10,000, antibacterial components such as quaternary ammonium compounds, salts of phenylmercuric known to have cold sterilization properties and to be harmless in use, thimerosal, benzalkonium chloride, methylparaben and propylparaben, benzyldodecylammonium bromide (benzyldodecinium bromide), benzyl alcohol, phenylethyl alcohol, buffering ingredients such as sodium chloride, sodium borate, sodium acetate or gluconate buffers and other conventional ingredients such as sorbitan monolaurate, triethanolamine, polyoxyethylene sorbitan monopalmitate, dioctyl sodium sulfosuccinate, thioglycerol, thiosorbitol, ethylenediaminetetraacetic acid and the like. In addition, suitable ophthalmic vehicles may be employed as carrier media for purposes of the present invention, including conventional phosphate buffer vehicle systems, isotonic boric acid vehicles, isotonic sodium chloride vehicles, isotonic sodium borate vehicles and the like.

The formulation may also include a gum, such as gellan gum at a concentration of 0.1-2% by weight, so that the aqueous eye drop gel contacts the eye, thereby providing the advantages of a solid ophthalmic insert as described in U.S. patent No. 4,861,760.

The pharmaceutical formulation may also be in the form of a solid insert, such as described in U.S. Pat. nos.4,256,108; 4,160,452 and 4,265,874, which remain substantially intact after dispensing of the medicament; or a bioerodible insert that is soluble or disintegrates in tear fluid as described in U.S. patent No. 4,287,175 or EPO publication 0077261.

Other ophthalmic formulations and delivery devices are described in the following patents: U.S. Pat. No. 5

4,014,335；4,300,557；5,098,443；5,188,826；5,378,475；5,422,116；5,424,078；5,466,233；5,725,493；5,773,019；5,773,021；5,776,445；5,814,635；5,888,493；6,235,781；6,297,228；6,372,245；6,511,660；6,579,519；6,582,422；6,713,081；6,719,750；

And U.S. patent application publication 2002-0064513; 2003, 0232089 and 2005, 0234018.

Medicament box

Generally, the kits of the invention comprise a corticosteroid and/or an NsIDI. These compounds may be provided in a kit as separate compositions or combined into a single composition. The kit of the invention may also contain instructions for administration of the corticosteroid and the NsIDI.

The kit of the invention may also contain instructions for administration of additional pharmaceutically acceptable substances (e.g., dipivefrin, anti-VEGF therapeutic, photodynamic therapy, NSAIDs, antiallergic agents, antihistamines, glaucoma therapeutic, artificial tears, antibiotics, antiviral agents, and antifungal agents) with corticosteroid and/or NsIDI. The kit may comprise any combination of corticosteroid, NsIDI, and additional pharmaceutically acceptable substances (i.e., any 1,2, or 3 of the above compounds).

Dosage form

The dosage of each compound in the claimed combination depends on several factors, including the method of administration, the disease to be treated, the severity of the disease, whether the disease is being treated or prevented, and the age, weight and health of the patient to be treated. In addition, pharmacogenomic (the effect of genotype on the pharmacokinetic, pharmacodynamic or efficacy properties of a treatment) information about a particular patient can affect the dosage used.

Continuous daily administration of the combination of the invention may not be required. The treatment regimen may require a cycle during which no medication is given, or may provide treatment on an as needed basis during acute inflammation.

As mentioned above, the compounds in question may be administered orally in the form of tablets, capsules, elixirs or syrups, or rectally in the form of suppositories. The compounds may also be administered topically in the form of foams, lotions, drops, ointments, creams, emollients or gels. Parenteral administration of the compounds is suitably carried out, for example, in the form of a saline solution or with the compound incorporated into liposomes. In cases where the compound itself is not sufficiently soluble, a solubilizing agent such as ethanol may be employed.

Conjugates

If desired, the drugs used in any of the combinations described herein can be covalently linked to each other to form conjugates of formula (VII)

(A)-(L)-(B)(VII)

In formula (VII), (a) is "drug 1" listed in table 1A or table 1B, covalently linked to (B) by linker (L), i.e. the corresponding "drug 2" listed in table 1A or table 1B.

Conjugates of the invention may be administered to a patient by any route to treat any of the diseases described herein.

The conjugates of the present invention may be prodrugs that release drug (a) and drug (B), for example, by cleavage of the conjugate by intracellular and extracellular enzymes (e.g., amidases, esterases, and phosphatases). Conjugates of the invention may also be designed to maintain integrity in vivo in large numbers, resisting the splitting action of intracellular and extracellular enzymes. Degradation of the conjugate in vivo can be controlled by designing the linker (L) and the covalent bond formed by drug (a) and drug (B) during synthesis of the conjugate.

Conjugates can be prepared using techniques familiar to those skilled in the art. Conjugates can be prepared, for example, using the methods disclosed in g.hermanson, Bioconjugate Techniques, academic press, 1996. The synthesis of conjugates may involve selective protection and deprotection of the alcohol, amine, ketone, thiol or carboxyl functions of drug (a), linker and/or drug (B). For example, commonly used amine protecting groups include carbamates such as t-butyl, benzyl, 2,2, 2-trichloroethyl, 2-trimethylsilylethyl, 9-fluorenylmethyl, allyl, and m-nitrophenyl. Other commonly used amine protecting groups include amides such as formamide, acetamide, trifluoroacetamide, sulfonamide, trifluoromethanesulfonamide, trimethylsilylethanesulfonamide, and tert-butylsulfonamide. Examples of commonly used protecting groups for carboxyl groups include esters such as methyl ester, ethyl ester, tert-butyl ester, 9-fluorenylmethyl ester, 2- (trimethylsilyl) ethoxymethyl ester, benzyl ester, diphenylmethyl ester, o-nitrobenzyl ester, orthoesters, and halogenated esters. Examples of commonly used protecting groups for alcohols include ethers such as methyl ether, methoxymethyl ether, methoxyethoxymethyl ether, methylthiomethyl ether, benzyloxymethyl ether, tetrahydropyranyl ether, ethoxyethyl ether, benzyl ether, 2-naphthylmethyl ether, o-nitrobenzyl ether, p-methoxybenzyl ether, 9-phenylchloroheteroanthryl ether, trityl (including methoxy-trityl) ether and silyl ether. Examples of commonly used protecting groups for mercapto groups include many of the same protecting groups as those for hydroxy groups. Furthermore, the thiol group may be protected in a reduced form (e.g. as a disulfide) or an oxidized form (e.g. as a sulfonic acid, sulfonate or sulfonamide). The protecting groups may be selected such that the selected conditions (e.g., acidic conditions, basic conditions, catalysis by nucleophiles, catalysis by lewis acids, or hydrogenation) require removal of every, unique other protecting group in the molecule. The conditions required for addition of protecting groups to amine, alcohol, thiol and carboxyl functions and for removal of these protecting groups are provided in detail in t.w.green and p.g.m.wuts, protecting groups in organic synthesis (second edition), john wiley & Sons, 1991 and p.j.kocienski, protecting groups, Georg Thieme Verlag, 1994. Additional synthetic details are provided below.

Joint

In the simplest case, the linker component of the invention is a bond between drug (a) and drug (B), but typically provides a linear, cyclic or branched molecular backbone having pendant groups that covalently link drug (a) to drug (B). Thus, the attachment of drug (A) to drug (B) is achieved by means of a covalent bond involving the formation of a bond with one or more functional groups located on drug (A) and drug (B). Examples of chemically reactive functional groups that may be used for this purpose include, but are not limited to, amino, hydroxyl, mercapto, carboxyl, carbonyl, glycosyl, vicinal diol, thioether, 2-aminoalcohol, 2-aminothiol, guanidino, imidazolyl, and phenolic groups.

Covalent attachment of drug (a) and drug (B) may be achieved using a linker comprising an active moiety that is capable of reacting with such functional groups present in drug (a) and drug (B). For example, the amine group of drug (a) may react with the carboxyl group of the linker or an activated derivative thereof, resulting in the formation of an amide bond linking the two.

Examples of moieties capable of reacting with a thiol group include XCH₂α -haloacetyl compounds of the type CO- (where X ═ Br, Cl or I), which moiety shows a specific reactivity with sulfhydryl groups, but which moiety can also be used to modify imidazolyl, thioether, phenol and amino groups, as in Gurd, Methods enzymol.11: 532 (1967). It is also believed that the N-maleimide derivative is selective for a thiol group, but that the N-maleimide derivative may be used for coupling to an amino group under certain conditions. Reagents such as 2-iminothiolane (Traut et al, Biochemistry 12: 3266(1973)) introduce a thiol group through the conversion of an amino group, which can be considered as thiol reagents provided that the linkage is created by the formation of a disulfide bridge.

Examples of reactive moieties capable of reacting with an amino group include, for example, alkylating agents and acylating agents. Representative alkylating agents include:

(i) alpha-haloacetyl compounds which show specificity for amino groups in the absence of reactive thiol groups and which belong to the group XCH₂CO- (where X ═ Cl, Br or I) type, for example as in WongBiochemistry 24: 5337(1979) the；

(ii) N-maleimide derivatives which can be reacted with amino groups by Michael type reactions or by acylation reactions with addition of a cyclic carboxyl group, such as, for example, Smyth et al, j.am.chem.soc.82: 4600(1960) and biochem.j.91: 589 (1964);

(iii) aryl halides, such as activated nitrohaloaromatic compounds;

(iv) alkyl halides, such as, for example, McKenzie et al, j. protein chem.7: 581 (1988);

(v) aldehydes and ketones capable of forming Schiff's base with amino groups, typically by reduction to produce stable amines, to stabilize the formed adducts;

(vi) epoxide derivatives such as epichlorohydrin and dioxirane, which are reactive with amino, mercapto or phenolic hydroxyl groups;

(vii) chlorine-containing derivatives of s-triazine, which are strongly reactive towards nucleophiles such as amino, mercapto and hydroxyl groups;

(viii) aziridines based on the above detailed s-triazine compounds, for example as described by Ross, j.adv.cancer res.2: 1(1954) by ring-opening with nucleophiles such as amino groups;

(ix) such as Tietz, chem. Ber.124: diethyl squarate as described in (1215), (1991); and

(x) α -haloalkyl ethers, such as Benneche et al, Eur.J.Med.chem.28: 463(1993), which is an alkylating agent more active than general alkyl halides because of activation by an ether oxygen atom.

Representative amino-reactive acylating agents include:

(i) isocyanates and isothiocyanates, especially aromatic derivatives, which form stable urea and thiourea derivatives, respectively;

(ii) sulfonyl chlorides, which have been prepared by Herzig et al, Biopolymers 2: 349 (1964);

(iii) an acid halide;

(iv) active esters such as nitrophenyl ester or N-hydroxysuccinimide ester;

(v) anhydrides such as mixed, symmetrical or N-carboxy anhydrides;

(vi) other useful reagents for amide bond formation, for example as described in M.Bodansky, Principles of Peptide Synthesis, Springer-Verlag, 1984;

(vii) acyl azides, for example, where sodium nitrite is used to generate the azide group from a preformed hydrazide derivative, such as Wetz et al, anal. biochem.58: 347 (1974); and

(viii) imidoesters, which react with amino groups to form stable amidines, for example as specified by Hunter and Ludwig, j.am.chem.soc.84: 3491 (1962).

Aldehydes and ketones can be reacted with amines to form Schiff's bases, which can be stabilized, preferably by reductive amination. The alkoxyamino moiety readily reacts with ketones and aldehydes to form stable alkoxyamines, such as, for example, Webb et al, Bioconjugate chem.1: 96 (1990).

Examples of reactive moieties capable of reacting with a carboxyl group include diazo compounds such as diazoacetate and diazoacetamide, which react with high specificity to produce an ester group, for example as described in Herriot, adv. 169 (1947). Carboxyl modifying reagents such as carbodiimides, which react by formation of O-acylureas followed by amide bond formation, can also be used.

It is understood that functional groups in drug (a) and/or drug (B) can be converted to other functional groups prior to reaction, if desired, e.g., to impart additional reactivity or selectivity. Examples of methods that can be used for this purpose include the conversion of amines to carboxyl groups using reagents such as dicarboxylic anhydrides; converting amines to thiols using reagents such as N-acetyl homocysteine thiolactone, S-acetyl mercaptosuccinic anhydride, 2-iminothiolane, or thiol-containing succinimidyl derivatives; converting thiols to carboxyl groups using reagents such as α -haloacetates; converting thiols to amines using reagents such as ethyleneimine or 2-bromoethylamine; conversion of the carboxyl group to an amine using a reagent such as a carbodiimide followed by a diamine; and converting the alcohol to a thiol using a reagent such as tosyl chloride, followed by transesterification with thioacetate and hydrolysis with sodium acetate to the thiol.

The so-called zero-length linker relates to a direct covalent attachment of the active chemical group of drug (a) to the active chemical group of drug (B) without introducing further linking substances, which linker can be used according to the invention if desired.

Most typically, however, the linker will comprise two or more reactive moieties as described above connected by a spacer element. The presence of such a spacer allows the bifunctional linker to react with the specific functional groups in drug (a) and drug (B), creating a covalent bond between the two. The active moieties in the linker may be the same (homobifunctional linker) or different (heterobifunctional linker, or heteropolyfunctional linker in which several different active moieties are present), providing a variety of potential agents that can cause covalent attachment between drug (a) and drug (B).

The spacer element in the joint is typically composed of straight or branched chains and may comprise C_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_2-6Heterocyclic group, C_6-12Aryl radical, C_7-14Alkylaryl group, C_3-10Alkylheterocyclic group or C_1-10A heteroalkyl group.

In some cases, the linker is described by formula (VIII):

G¹-(Z¹)_o-(Y¹)_u-(Z²)_s-(R₃₀)-(Z³)_t-(Y²)_v-(Z⁴)_p-G²(VIII)

in formula (VIII), G¹Is between the drug (A) and the linkerA bond between; g²Is a bond between the linker and the drug (B); z¹、Z²、Z³And Z⁴Each independently selected from O, S and NR₃₁；R₃₁Is hydrogen, C_1-4Alkyl radical, C_2-4Alkenyl radical, C_2-4Alkynyl, C_2-6Heterocyclic group, C_6-12Aryl radical, C_7-14Alkylaryl group, C_3-10Alkylheterocyclic group or C_1-7A heteroalkyl group; y is¹And Y²Each independently selected from carbonyl, thiocarbonyl, sulfonyl or phosphoryl; o, p, s, t, u and v are each independently 0 or 1; and R₃₀Is C_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_2-6Heterocyclic group, C_6-12Aryl radical, C_7-14Alkylaryl group, C_3-10Alkylheterocyclic group or C_1-10Heteroalkyl radicals or radicals G¹-(Z¹)_o-(Y¹)_u-(Z²)_s-and- (Z)³)_t-(Y²)_V-(Z⁴)_P-G²A chemical bond of attachment.

Examples of homobifunctional linkers useful in preparing conjugates of the invention include, but are not limited to, diamines and diols selected from the group consisting of: ethylene diamine, propylene diamine and hexamethylene diamine, ethylene glycol, diethylene glycol, propylene glycol, 1, 4-butanediol, 1, 6-hexanediol, cyclohexanediol and polycaprolactone diol.

Further applications

The compounds of the invention may be used in immunomodulatory or mechanistic assays to determine whether other combinations or single agents are as effective as the combination in inhibiting the secretion or production of proinflammatory cytokines or modulating an immune response using assays generally known in the art, examples of which are described herein. After a suitable time, the cells are examined for cytokine secretion or production or other suitable immune response. Combinations were compared to each other with respect to the relative effects of the individual drugs, and potent compounds and combinations were identified.

The combination of the invention is also a useful tool for elucidating mechanistic information about biological pathways involved in inflammation. Such information may lead to the development of new combination drugs or single drugs that inhibit inflammation caused by pro-inflammatory cytokines. Methods for determining biological pathways known in the art can be used to determine pathways or pathway networks affected by contacting cells that stimulate the production of proinflammatory cytokines with a compound of the invention. Such methods may include assaying for cellular components that are expressed or inhibited following contact with a compound of the invention, or assaying for some other metabolic activity of the cell, such as enzymatic activity, nutrient uptake and proliferation, as compared to untreated, positive or negative control compounds and/or novel single and combination drugs. Cellular components analyzed may include gene transcription and protein expression. Suitable methods may include standard biochemical techniques, radiolabeling of the compounds of the invention (e.g.¹⁴C or³H-labeling) and observing the compounds bound to the protein, e.g., using a 2d gel, gene expression profile. Once identified, such compounds can be used in vivo models to further validate tools or develop new anti-inflammatory agents.

The following examples serve to illustrate the invention. They are not intended to limit the invention in any way.

Example 1

The effect of the test compound combinations on TNF α secretion in leukocytes from human buffy coats stimulated with lipopolysaccharide or phorbol 12-myristate 13-acetate (PMA) and ionomycin was determined as follows. The results of these tests are described in FIGS. 1A-1 MM.

Lipopolysaccharide (LPS)

100 μ L of diluted human leukocyte suspension contained in each well of a polystyrene 384-well plate (NalgeNunc) was stimulated to secrete TNF α by treatment with lipopolysaccharide (Sigma L-4130) at a final concentration of 2 μ g/mL. Different concentrations of each test compound were added at the time of stimulation. After incubation at 37 ℃ for 16-18 hours in a humidified incubator, the plates were centrifuged and the supernatant was transferred to white opaque polystyrene 384-well plates (NalgeNunc, Maxisorb) coated with anti-TNF α antibody (PharMingen, # 551220). After 2 hours of incubation, the plates were washed with PBS containing 0.1% tween 20 (Tecan powerwash 384), incubated for an additional 1 hour with another anti-TNF α antibody labeled with biotin (PharMingen, #554511) and HRP conjugated to streptavidin (PharMingen, # 13047E). After washing the plates with 0.1% tween 20/PBS, HRP-luminescent substrate was added to each well and light intensity was measured using a LJL Analyst plate luminometer.

PMA/ionomycin

100 μ L of diluted human leukocyte suspension contained in each well of a polystyrene 384 well plate (NalgeNunc) was stimulated to secrete TNF α by treatment with phorbol 12-myristate 13-acetate (Sigma, P-1585) and ionomycin (Sigma, I-0634) at a final concentration of 10 ng/mL. Different concentrations of each test compound were added at the time of stimulation. After incubation at 37 ℃ for 16-18 hours in a humidified incubator, the plates were centrifuged and the supernatant was transferred to white opaque polystyrene 384-well plates (NalgeNunc, Maxisorb) coated with anti-TNF α antibody (PharMingen, # 551220). After 2 hours of incubation, the plates were washed with PBS containing 0.1% tween 20 (Tecan powerwash 384), incubated for an additional 1 hour with another anti-TNF α antibody labeled with biotin (PharMingen, #554511) and HRP conjugated to streptavidin (PharMingen, # 13047E). After washing the plates with 0.1% tween 20/PBS, HRP-luminescent substrate was added to each well and light intensity was measured using a LJL Analyst plate luminometer.

Example 2

Both corticosteroids and NsIDI inhibit cytokine production in cell culture models of immune function. We tested the effect of different concentrations of the combination of NsIDI and corticosteroid on cytokine production in a cell culture model of immune function. We propose that combinations exhibiting synergistic or additive effects can be used to treat ocular diseases at concentrations low enough to avoid undesirable side effects.

Measuring inhibitory Activity of proinflammatory cytokines

The inhibition of IFN γ, IL-2 and TNF α by the compound dilution medium was determined as described below. The results of these experiments are shown in FIGS. 2A-2 LL.

IFNγ

100 μ L of diluted human leukocyte suspension contained in each well of a polystyrene 384 well plate (NalgeNunc) was stimulated to secrete IFN γ by treatment with phorbol 12-myristate 13-acetate (Sigma, P-1585) and ionomycin (Sigma, I-0634) at a final concentration of 10 ng/mL. Different concentrations of each test compound were added at the time of stimulation. After incubation at 37 ℃ for 16-18 hours in a humidified incubator, the plates were centrifuged and the supernatant was transferred to a white opaque polystyrene 384 well plate (NalgeNunc, Maxisorb) coated with anti-IFN γ antibody (Endogen, # M-700A-E). After 2 hours of incubation, the plates were washed with Phosphate Buffered Saline (PBS) containing 0.1% tween 20 (polyoxyethylene sorbitan monolaurate) (tecanpowerwash 384), incubated with another anti-IFN γ antibody labeled with biotin (Endogen, M701B) and horseradish peroxidase (HRP) conjugated to streptavidin (PharMingen, #13047E) for an additional 1 hour. After washing the plates with 0.1% tween 20/PBS, HRP-luminescent substrate was added to each well and light intensity was measured using a LJL Analyst plate luminometer.

IL-2

mu.L of a diluted human leukocyte suspension contained in each well of a polystyrene 384 well plate (NalgeNunc) was stimulated to secrete IL-2 by treatment with phorbol 12-myristate 13-acetate (Sigma, P-1585) and ionomycin (Sigma, I-0634) at a final concentration of 10 ng/mL. Different concentrations of each test compound were added at the time of stimulation. After incubation at 37 ℃ for 16-18 hours in a humidified incubator, the plates were centrifuged and the supernatant was transferred to white opaque polystyrene 384-well plates (NalgeNunc, Maxisorb) coated with anti-IL-2 antibody (PharMingen, # 555051). After 2 hours of incubation, the plates were washed with PBS containing 0.1% Tween 20 (Tecan PowerWasher 384), incubated for an additional 1 hour with another anti-IL-2 antibody labeled with biotin (Endogen, M600B) and HRP conjugated to streptavidin (PharMingen, # 13047E). After washing the plates with 0.1% tween 20/PBS, HRP-luminescent substrate was added to each well and light intensity was measured using a LJL Analyst plate luminometer.

TNFα

The effect of the combination of test compounds on TNF α secretion in leukocytes from human buffy coat stimulated with phorbol 12-myristate 13-acetate was determined as follows. Human leukocytes from buffy coat were diluted 1: 50 in medium (RPMI; Gibco BRL, # 11875-. Add drug to the indicated concentration. In a humidified incubator at 37 ℃ and 5% CO₂After incubation for 16-18 hours under conditions, the plates were centrifuged and the supernatant was transferred to a white opaque polystyrene 384 well plate (NalgeNunc, Maxisorb) coated with anti-TNF α antibody (PharMingen, # 551220). After 2 hours of incubation, the plates were washed with PBS containing 0.1% tween 20 (Tecan powerwash 384), incubated with biotin-labeled anti-TNF α antibody (PharMingen, #554511) and streptavidin-conjugated HRP (PharMingen, #13047E) for an additional 1 hour. Plates were then re-washed with 0.1% Tween 20/PBS. HRP-emitting substrate was added to each well and the light intensity of each well was measured using a plate luminometer.

Percent inhibition

Percent inhibition (% I) per well was calculated using the following formula:

% I ═ 100 [ (average untreated pores-treated pores)/(average untreated pores) ]

The value of the mean untreated well (mean untreated well) is the arithmetic average of 40 wells of the same assay plate treated with vehicle alone. Negative inhibition values were obtained from the local variance in the treated wells compared to the untreated wells.

Preparation of the Compounds

Stock solutions containing cyclosporin A were prepared at a concentration of 1.2mg/ml in DMSO. Stock solutions of tacrolimus were prepared at a concentration of 0.04mg/ml with DMSO. Stock solutions containing corticosteroids were prepared at final concentrations of 0-40 μ M with dimethyl sulfoxide (DMSO). Master plates were prepared containing dilutions of the above compound stock solutions. The master plate was sealed and stored at-20 ℃ for future use.

The final single drug plate was generated by transferring 1. mu.L of stock solution from a particular master plate using a Packard Mini-Trak liquid handler into a dilution plate containing 100. mu.L of medium (RPMI; Gibco BRL, # 11875-. The dilution plates were then mixed, 5mL aliquots were transferred to the final assay plates, which were pre-filled with 50 mL/well of RPMI medium containing the appropriate stimulating agent to activate secretion of IFN γ, IL-2 or TNF α.

Other embodiments

Various modifications and variations of the methods and compositions of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been described in connection with specific desired embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described methods of carrying out the invention which are obvious to those skilled in the fields of medicine, immunology, pharmacology, endocrinology or related fields are intended to be within the scope of the invention.

All publications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

Claims (5)

1. A composition suitable for ocular administration comprising prednisolone or prednisolone acetate and cyclosporin a, wherein the prednisolone or prednisolone acetate is present at a concentration of 0.12% and the cyclosporin a is present at a concentration of 0.01% or 0.02%.

2. The composition of claim 1, wherein said cyclosporin a is present at a concentration of 0.01%.

3. The composition of claim 1, wherein said cyclosporin a is present at a concentration of 0.02%.

4. The composition of claim 1, wherein the composition is a solution, gel, cream, suspension, or emulsion.

5. The composition of claim 1, wherein said prednisolone or prednisolone acetate and said cyclosporin a are the only active ingredients.