• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The present invention provides 3-deoxyflavonoid compounds, and methods of inhibiting T-cell activity and treating diseases and disorders (eg, autoimmune disorders, inflammatory disorders, diabetes mellitus, ALS, MS, rheumatoid arthritis, etc.). . In some cases, the efficacy and / or time of action of luteolin and / or other 3-deoxyflavonoid compounds can be increased by administering such compounds with rutin, rutin, and / or rutin derivatives. Also, in some cases, the first pass metabolism of luteolin or other 3-deoxyflavonoids can be prevented by administering such compounds by parenteral routes (eg, sublingual, buccal, intranasal, injection, etc.).
    公开号:KR20040048407A
    申请号:KR10-2004-7003414
    申请日:2002-09-06
    公开日:2004-06-09
    发明作者:토마스 피. 라헤이；브이. 제이. 라자드햐크샤
    申请人:시녹스, 인크.；
    IPC主号:
    专利说明:

    Inhibition by 3-Deoxyflavonoids of T-Lymphocyte Activation and Therapies Related Thereto}
    [1] <Related application>
    [2] This application is directed to U.S. Provisional Application No. 60 / 317,666, filed on September 6, 2002 and filed on August 30, 2002, entitled "Oral of 3-deoxyflavinoids to avoid First Pass Metabolism. Priority is claimed for US provisional application titled "Administration" (serial number will be given), the entire contents of which are incorporated herein by reference.
    [4] Flavonoids are polyphenolic compounds present everywhere in foods of plant origin. Over 4000 structurally unique flavonoids have been identified in plant sources [Harborne et al., 1975 The Flavonoids, Academic Press, New York; Cody V, Middleton E, Harborne JB and Beretz A eds; Alan R. Liss, Inc, New York, 1986 Plant Flavonoids in Biology nad Medicine, Part 1 and 2]. Flavonoids are present in fruits, vegetables, nuts, berries, herbs, spices, stems, and flowers, as well as in tea and red wine, and are the main constituents of citrus fruits and other food ingredients, and are consumed as a common human diet.
    [5] Flavonols not claimed in the present invention are the most abundant natural flavonoids, the content of which in the most common edible fruits, vegetables and berries can reach hundreds of mg per kg of dry weight. Initial estimates showed that the average daily absorption of flavonoids totaled about 1.0 g and 115 mg of flavonol and flavone fractions. Recently, a "Seven Countries Study" found that the total daily uptake of flavonoids could vary from 2.6 to 68.2 mg, with the fraction of quercetin between 39 and 100%. Another study of 17 volunteers from 14 countries found that the average consumption of quercetin and camphorol was approximately 28 mg / day [Makris and Rossiter, 2001 J. Agric. Food Chem., 49, 3216.
    [6] Since most investigations of the effects of flavonoid-containing vegetable foods have been based on analytical data on raw plant tissues, they merely represent the composition of the foods in the raw state. Environmental variables and processing will significantly affect the concentration and biological activity of the flavonoids, which are not considered. A rarely done recent study of the effects of home and plant processing practices on flavonoid composition of plant foods has shown that treatments commonly performed at home, such as boiling, frying, and microwave cooking, reduce quercetin concentrations in onions and tomatoes. To lower than 82%. Furthermore, it was found that the blanching reduced the amount of quercetin and camphorol in onions to 39 and 64%, respectively, and the mycetin and quercetin in sweet potatoes to be 19 and 50%, respectively. Nevertheless, this aspect is very important considering that only a small amount of fruits and vegetables are consumed in raw form, but most of them need to be processed for safety, quality and economic reasons.
    [7] Estimating the total absorption of flavonoids is difficult because only limited data are available for food ingredients. Several efforts have been made to quantify the amount of different flavonoids in various kinds of food plants. Hertog et al. (1992) J Agric Food Chem 40: 2379-2383, the average daily uptake of mixed flavonoids is only 23 mg / year based on data from the Dutch National Food Consumption Survey from 1987-88. It was work. Flavonoids measured were quercetin, camphorol, myricetin and 3-deoxyflavone, apigenin and luteolin, 3-hydroxy flavones. The absorption of these five antioxidant flavonoids is 23 mg / day, which exceeds the absorption of other well-known antioxidants such as β-carotene (2-3 mg / day) and vitamin E (7-10 mg / day). About one third of the average absorption of vitamin C (70-100 mg / day). The amount of 23 mg / day was usually flavonol and flavone, measured as aglycone, with the majority of flavonoids consumed (16 mg / day) quercetin. However, it should be emphasized that recent evidence points out that flavonoid-glycosides are absorbed much more easily by humans (compared to aglycones) [Hollman and Katan, 1998 Arch Toxicol Suppl 20: 237-248 and Absorption, metabolism , and bioavailability of flavonoids, in Flavonoids in Health and Disease (Rice-Evans CA and Packer Leds, 1998 pp 483-522, Marcel Dekker, Inc., New York). Moreover, the amounts and ingredients vary considerably from country to country, and flavonoids can provide physiologically significant concentrations in body fluids and tissues, probably in most developed countries, except for Mediterranean coastal foods, which are rich in olive oil, citrus fruits and vegetables. In particular, there will be a shortage of foods rich in 3-deoxyflavonoids.
    [8] Flavonoids are typical phenolic compounds and therefore act as strong metal chelators and free radical scavengers and are powerful chain-cutting antioxidants. Because of their antioxidant properties, they have some beneficial health effects. Flavonoids have a number of notable biochemical and physiological actions, some of which suggest that certain members of this group of compounds can significantly affect the various cellular system functions of mammals. They have long been recognized as having anti-inflammatory, antioxidant, anti-allergic, hepatoprotective, antithrombotic, antiviral and anticarcinogenic activity. However, Rimm and his co-workers [1996 Ann Intern Med 125: 384-389] did not reveal a strong inverse relationship between the absorption of flavonoids and all coronary heart disease. The uptake of flavonols and flavones was inversely correlated with the accompanying coronary artery disease in most but not all prospective epidemiological studies.
    [9] Vegetables and fruits, which are often associated with low cancer incidence in epidemiological studies, are not the main sources of dietary flavonols and flavones, so they will not be a direct cause of cancer prevention effects, and their preventive effects on cardiovascular disease are also not critical. It has also been established in the literature that quercetin, flavonol (3-hydroxyflavone), is mutagenic and therefore its glycoside rutin is expected to behave similarly due to easy hydrolysis to quercetin. The inventors have unexpectedly found that flavonols may have the opposite effect as the three-deoxyflavonoids of the present invention and may interfere with the therapeutic benefits of 3-deoxyflavonoids.
    [10] Flavonoids present in foods were considered non-absorbent because they were bound to sugars as β-glucosides. However, the human uptake (52%) of quercetin glycosides from onions is known to be much better than that of pure aglycone (24%) found by the Dutch researchers. More recent analytical techniques were used to measure plasma quercetin concentrations after digestion of fried onions containing 64 mg of quercetin aglycone and an equivalent quercetin glycoside [Hollman et al., 1996 Free Radical Biol Med 21: 703-707 ]. A plasma maximum concentration of 196 μg / ml was achieved after 2.9 hours with an absorption half-life of 0.87 hours. The half life of the dispensing step was 3.8 hours and the half life of the elimination step was 16.8 hours. Thus, quercetin from food (cooked vegetables) may be absorbed through the mouth to reach tissues and plasma where antioxidants and other activities can be exerted. Quercetin corresponds to other flavonoids in other plant sources. Hallman and Katan (1998 Arch Toxicol Suppl 20: 237-248) examined the bioavailability and health effects of food flavonol in humans. They found that quercetin glycosides in onions were more readily absorbed than pure aglycones, and that the absorbed quercetin was slowly removed from the blood, suggesting that the gut-hepatic circulation could be operable. In a related study, Holman et al. [1995 FEBS Lett 418: 152-156] concluded that quercetin-glucose conjugates could be more readily absorbed and suggested that glycosides could be absorbed through the intestinal sugar absorption pathway. . Digested flavonoid glycosides are rapidly converted to metabolites into aglycones by ubiquitous glycosidase, and it is evident from the prior art that pharmacokinetic parameters are dramatically changed due to poor solubility of aglycones. Oral absorption and bioavailability of aglycone are important issues in the treatment of diseases. The formulations described in the present invention utilize absorption enhancers that also act as solubilizers and are expected to overcome this problem by facilitating the absorption of aglycone. Moreover, many of the 3-deoxyflavonoids of formula I of the present invention are designed to significantly increase water solubility compared to corresponding aglycones, and they are expected to exhibit very improved pharmacokinetic profiles.
    [11] The complex nature of plants and herbs presents some special challenges to challenge. Plants typically consist of multiple components, some of which may be active alone or in combination with each other. Such multiple actives may exhibit agonist or antagonist activity even though they have a common structural backbone. The identification of bioactive components is of vital importance in solving bioavailability problems and in establishing meaningful criteria for dissolution, bioavailability, active ingredients, uniformity and stability.
    [12] The complex relationship of plant activity-composition presents a unique problem for understanding the effects of formulation and processing variables on product quality, establishing good manufacturing practices and ensuring that appropriate quality and performance criteria are met [Augsburger, 2001] in Examining the Science behind Nutraceuticals, AAPS Press]. Environmental variables and processing can significantly affect concentration and biological activity.
    [13] Another approach to this problem is to identify and synthesize the active ingredients in their purest form to remove unwanted ingredients that may be undesirable or lack biological activity.
    [14] These low molecular weight substances found in all vascular plants are phenylbenzo-pyrones (phenylchromones), which are classified into structures based on a common tricyclic nucleus. They are usually subdivided into flavanols, anthocyanidins, flavones, flavanones and chalcones depending on the substituents. The basic flavone and flavanone structure consists of two benzene rings (A and B) linked in the center via a heterocyclic pyron ring (Formula I). The 3-deoxyflavonoid compounds of the invention are in particular based on the presence (or absence) of double bonds between the carbon atoms 2 and 3 of the pyran ring and the absence of hydroxyl groups at the 3-position of the central pyron ring (Formula I). Shall be. In the flavonoid structure, the phenyl group is usually substituted at the 2-position of the pyron ring. In isoflavonoids substituted at the 3-position, genistein and daidzein are included in the compositions of the present invention.
    [15] Diabetes and autoimmune diseases affect nearly 10% of the world's population at the same time. Autoimmune diseases are associated with the regulation of immune abnormalities mediated by cells and body fluids, and are often associated with abnormal or increased T cell, B cell and macrophage agonist action induced by autoantigens. Activation of these cellular components on autoantigens is thought to be associated with the destruction of feedback mechanisms associated with autoimmune resistance. Autoimmune diseases include a wide range of clinical diseases and many similarities, although different target organs (Ahmed et al., Am J, Path., 121: 531 (1985)). In addition, these diseases are all chronic, and due to insufficiently known causes, clinical symptoms tend to relieve and recur, and other organs are involved. Although the presence of autoantibodies, inadequate expression of class II antigens, activation of macrophages and penetration of T cells into target organs have been found in essentially all autoimmune diseases, the mechanisms leading to disease activation or disease progression are well known. Not. Thus, therapies for these diseases are often unsatisfactory and include the use of gold salts, methotrexate, antimalarial agents, glucocorticoids (methylprednisolone), beta interferon, and other immunosuppressive agents, as well as attempts to induce plasma export and immunotolerance. do. Therapies for autoimmune diseases have not improved much over the past decade and are mainly associated with the use of nonsteroidal and steroidal anti-inflammatory drugs to treat the symptoms of the disease. While it is clearly necessary to suppress certain immune responses induced against the host, generalized immunosuppression with glucocorticoids tends to have adverse side effects profiles and the tendency that immunosuppressed patients are more at risk for other infectious and non-infectious diseases. It has major obstacles in that it is.
    [16] The role of estrogen in disease progression or degeneration is complex and depends on the nature of the autoimmune disease, but estrogen appears to be associated with autoimmune disease. For example, estrogens appear to have the effect of relieving rheumatoid arthritis while exacerbating systemic lupus [Chander &Spector; Ann. Rheum. Dis. 50: 139. Estrogens have been shown to have an inhibitory role on T cell function but have an immunostimulatory effect on B cells. Thus, estrogen-like compounds should prove beneficial in diseases associated with activated T cells, including rheumatoid arthritis, multiple sclerosis, Guillain-Barre syndrome, and Hashimoto's thyroiditis by inhibiting T cell function [Holmadahl, J. Autoimmun. 2: 651 (1989).
    [17] Glucocorticoids and other immunosuppressive medications such as cyclophosphamide (CPA) are critically important for the survival of patients suffering from systemic lupus erythematosus. There is no specific cure yet. To date, treatment has been aimed at preventing or overcoming acute exacerbations and avoiding relapses. To this end, patients have been treated with glucocorticoids and other immunosuppressants, but they themselves have deleterious side effects.
    [18] Angiogenesis-dependent diseases (ie, those that require or induce vascular growth) make up a significant portion of all diseases that seek medical treatment. Cancer, for example, is the second leading cause of death in the United States, accounting for more than one fifth of all deaths. In short, cancer is most typically characterized by uncontrolled division of a population of cells that form one or more tumors. These tumors are also characterized by internal growth of the vasculature, which provides several factors that enable continued tumor growth. Cancer is generally easier to diagnose than in the past, but many forms are still incurable, even if found early.
    [19] Currently, various methods have been used to treat cancer, including various surgeries, for example. However, if treated with surgery alone, many patients (especially those suffering from certain types of cancer, such as breast cancer, brain cancer, colon cancer and liver cancer) will experience cancer recurrence. Thus, many cancers are also treated in combination with therapy and / or radiation therapy using cytotoxic chemotherapeutic drugs (eg, vincristine, vinblastine, cisplatin, methotrexate, 5-FU, etc.) in addition to surgery. However, one difficulty with this method is that radiotherapy and chemotherapeutic agents are toxic to normal tissue and often cause life-threatening side effects. Furthermore, these methods often have very high failure rates / remission rates.
    [20] Angiogenesis or angiogenesis is the growth and development of new arteries. This is important for the normal development of the vascular system, including damage-repair. However, there are symptoms characterized by abnormal angiogenesis, including diabetic retinopathy, neovascular glaucoma, rheumatoid arthritis, psoriasis and certain cancers. Diabetic retinopathy, for example, is the leading cause of blindness. Diabetic retinopathy is of two types: simplicity and proliferation. Proliferative retinopathy is characterized by angiogenesis and scar formation. About half of patients with proliferative retinopathy develop blindness within about 5 years. It would be desirable to find antiangiogenic agents useful for the treatment of these diseases. The present invention provides compositions and methods suitable for the treatment of cancer, and also other non-neoplastic angiogenesis-dependent diseases, and also provide other related advantages.
    [21] Autoimmune diseases include a wide range of clinical disorders in which the patient's immune system incorrectly attacks itself by targeting the cells, breakfast, and organs of the patient's own body. The following are some examples of autoimmune diseases that are classified for target organs that are primarily involved by each disease:
    [22] Nervous system: multiple sclerosis, myasthenia gravis, autoimmune neuropathy such as Guillain-Barré, autoimmune uveitis,
    [23] Blood: autoimmune hemolytic anemia, pernicious anemia, autoimmune thrombocytopenia,
    [24] Vascular system: temporal arteritis, antiphospholipid syndrome, vasculitis, such as Wegener's granulomatosis, Behcet's disease,
    [25] Skin: psoriasis, herpes dermatitis, vulgaris, vitiligo,
    [26] Gastrointestinal tract: Crohn's disease, ulcerative colitis, primary biliary cirrhosis, autoimmune hepatitis,
    [27] Endocrine system: Type 1 diabetes mellitus, Addison's disease, Graves' disease, Hashimoto's thyroiditis, autoimmune ovarian infection and testicles,
    [28] Multi-organ and / or musculoskeletal system: rheumatoid arthritis, systemic lupus erythematosus, scleroderma, polymyositis, dermatitis, such as ankylosing spondylitis, Sjogren's syndrome, interstitial cystitis.
    [29] Regardless of the specific organ affected, T-lymphocytes are believed to be the cause of the development of autoimmune diseases. Therapies currently used for these diseases are often unsatisfactory and are typically glucocorticoids (eg, methylprednisolone, prednisone), nonsteroidal anti-inflammatory agents, gold salts, methotrexate, antimalarial agents, and other immunosuppressive agents such as cyclosporin and FK-506 Use Unfortunately, these T-cell inhibitory drugs are toxic and have liver and kidney toxicity that limits their use.
    [30] Thus, the study of additional immunosuppressive agents for the treatment of autoimmune and inflammatory diseases has attracted considerable interest in the pharmaceutical industry. Since cytokines such as interferon-gamma and tumor necrosis factor-alpha play an important role in the pathophysiology of autoimmune diseases, they are involved in the development of agents that inhibit their production, secretion and / or end-organ effects. Much effort has been made.
    [31] There is an excellent record of progress in treating neurological and cardiovascular diseases using ion channel modulators (openers or blockers). Ion channel blockers of the general kind are the main therapeutics for the treatment of sleepiness, epilepsy and arrhythmia. Because ion channels play a major role in T-cell immune responses, these channels can be attractive targets for pharmaceutical immunomodulation.
    [32] Potassium (K + ) channels are found in all tissues of the body, and from homeostatic control of cell volume and osmotic balance to control of electrical signaling in nerve and muscle cells, transporters at nerve endings and hormone secretion in endocrine cells It has several roles ranging from regulation. To coordinate these various functions, there are about 80 genes encoding K + channels. Of all the ion channels, the K + channel phase is by far the most diverse. K ATP channels are important targets for antidiabetics that promote insulin release [Potassium Channels: Molecular Defects, Diseases, and Therapeutic Opportunities, Shieh CC et al, 2000 Pharmacol Rev, 52, 557-583 and references cited therein] literature].
    [33] Ca ++ mediated signaling events are important for the bioactivity of various cell types. Ca ++ - activated K + (K Ca) channels within a cell since the Ca ++ response to the opening according to the change in the ([Ca ++] i), by controlling the membrane potential in all excitable and non-excitable cells Ca + + Plays an important role in regulating signaling cascade In history, these channels have large versus (BK Ca ), medium ((IK Ca ) and small (SK Ca ) conductivity (100-250 pS, 11-40, respectively) based on signal-channel conductivity in symmetric K + solutions. pS and 4-14 pS). BK Ca channels, which are abundant in smooth muscle and neurons but also present in other cells, are opened by high [Ca ++ ] i as well as depolarization and are closed by the scorpion peptides caribidotoxin (ChTX) and iberiotoxin.
    [34] The SK Ca channel is very sensitive to [Ca ++ ] i and voltage independent for activation in the 200 to 500 nM range. SK Ca channels are highly expressed in the central nervous system, skeletal muscle and human Jurkat T-cells and are blocked by apamin, a peptide from bee venom, and by a scorpion peptide, silatoxin. Three genes in the novel subfamily (SKCa1-3) encode SK Ca channels.
    [35] Unlike the SK Ca channel, the IK Ca channel is mainly expressed in peripheral tissues including tissues of the hematopoietic system, colon, lung, placenta and pancreas. These channels can be pharmacologically distinguished from SK Ca channels by their sensitivity to blocking by ChTX and clotrimazole and their insensitivity to apamin. Both SK Ca and IK Ca channels are voltage independent and are steeply sensitive to the rise of [Ca ++ ] i . At least one gene called IKCa1 has been demonstrated to encode native IK Ca channels in human T-lymphocytes and red blood cells, and colon epithelium. Human IKCa1 shares only about 40% identity with the SKCa1-3 gene product and includes a distinct subfamily in the extended K + channel super-gene family.
    [36] In the beta cells of the pancreas, insulin secretion is triggered by elevated blood glucose that is metabolically bound to the closure of K ATP channels. When the K ATP channel is closed, the membrane potential is depolarized, resulting in activation of the voltage-gated Ca 2+ channel and controlled extracellular outflow of granules containing insulin. Sulfonylurea compounds such as glymepyride are used to treat type II diabetes to enhance insulin release from islets. These drugs work by blocking K ATP channels. In addition, islets express Kv1.7 channels and increased Kv1.7 mRNA levels are detected in islets of diabetic db / db rats (Kalman et al., 1998 J. Biol. Chem. 273: 5851). Recent studies have identified various KCa channels in islets, and SKCa3 has been demonstrated to regulate insulin secretion under several circumstances in transgenic mice (Tamarina et al., 2001 Biophysical Society Abstract 1472.) In addition, calcium in these cells Channels are required for insulin secretion A 3-deoxyflavonoid compound can regulate blood glucose by targeting one or more of these channels.
    [37] The dominant voltage gate channel in human T-lymphocytes is encoded by Kv1.3, a shaker related gene. Kv1.3 is widely characterized at the molecular and physiological levels and plays an important role in regulating T-lymphocyte proliferation mainly by maintaining the resting membrane potential of resting T-lymphocytes. Highly specific peptide blockers of this channel, such as margatoxin and ShK-Dap 22 (Kalman 1998, J. Biol. Chem. 273: 32697), have been described as stop T- to perform mitogen induced activation. Inhibit the ability of lymphocytes. T-lymphocyte activation in humans is performed with about a 2-fold increase in Kv1.3 current, while K Ca current is upregulated by 10-25 times (Zweifach 1993, Proc. Natl. Acad. Sci. USA 90: 6295 and Chandy 1993, Seminars in The Neurosciences 5: 125).
    [38] The early stages of T-cell activation can be conceptually separated into pre-Ca ++ and post-Ca ++ events. After binding of the antigen to the T-cell antigen-receptor, the activation of tyrosine kinase and the generation of inositol 1,4,5-triphosphate are characterized by the influx of Ca ++ through calcium-releasing activated calcium (CRAC) channels and cytoplasmic Ca ++. Results in an increase in concentration (Kerschbaum and Cahalan 1999 Science 283: 836 and references cited therein). The elevation of Ca ++ activates phosphatase calcineurin and then dephosphorylates cytoplasmic localized nuclear factors of activated T cells (NF-AT) and accumulates them in the nucleus to bind to promoter elements of the interleukin-2 gene. do. With parallel events involving activation of protein kinase C and ras, gene transcription results in lymphokine secretion and lymphocyte proliferation. Some genes require long lasting Ca ++ signals, while others require only a temporary increase in Ca ++ . Moreover, Ca ++ immobilization of T-cells at the antigen presenting site helps to enhance the interaction between T-cells and antigen presenting cells, thereby facilitating local signaling between cells. Preparation of the core T cell cytokine IL-2 requires simultaneous activation of both pathways, where Ca 2+ is absolutely required for the process.
    [39] Two distinct types of potassium channels (voltage gate Kv1.3 channel and intermediate conductivity calcium activated potassium channel IKCa1) indirectly determine the driving force of calcium influx through the storage-activated Ca ++ channel. When these potassium channels are open, the resulting effluent of K + hyperpolarizes the membrane to highlight the influx of Ca ++ which is absolutely required for downstream activation events (Cahalan and Chandy 1997, Curr. Opin. Biotechnol 8: 749)). Blockers of Kv1.3 and IKCa1 channels, when applied independently, inhibit human T-cell activation and, when applied together, produce greater inhibition. One mechanism for immunosuppression by K + channel blockers is through membrane depolarization, which reduces Ca ++ influx through CRAC channels in the T-cell membrane, and thus calcium dependent signaling during human T-cell activation. Suppress the incident Upregulation following T-lymphocyte activation, with its dominant expression in peripheral tissues, makes the IKCa1 channel a very interesting target for the development of novel immunosuppressive agents. The azole antifungal clotrimazole, the most specific available inhibitor of IKCa1, has recently been demonstrated to effectively inhibit T-lymphocyte proliferation (Khanna 1999, J. Biol. Chem. 274: 1483822). However, clotrimazole is also an effective inhibitor of many mammalian cytochrome P-450-mediated responses and is not an ideal therapeutic candidate. Triaryl methane analogs of 1-[(2-chlorophenyl) diphenylmethyl] -1H-pyrazole, clotrimazole have been shown to inhibit native IKCa1 channels cloned in human T-lymphocytes with a K d of 20 to 25 nM. Proven and 200 to 1500 fold selective over other ion channels (Wulff 2000, Proc. Natl. Acad. Sci. USA 97: 8151).
    [40] In view of the drawbacks associated with currently available therapies for autoimmune diseases, there remains a need for the development of novel immunosuppressive drugs that can selectively inhibit the activation of T-lymphocytes with minimal side effects.
    [41] Summary of the Invention
    [42] According to the present invention there is provided a method of inhibiting the activation or proliferation of T-lymphocytes in a patient by administering an inhibitory amount of T-lymphocytes of a compound of formula (I) to a human or veterinary patient.
    [43]
    [44] Wherein X is selected from O and S,
    [45] R 1 to R 5 and R 9 to R 12 are H, OH, halogen, eg F or Cl, alkyl, amino, NHMe, SH, Sme, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycar Bonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucoronide, and O-CO-A — (CH 2 ) n —NR′R ″ wherein A is phenyl or substituted phenyl, or n is 0 to 5, R ′ and R ″ are H, lower alkyl, hydroxyalkyl, aminoalkyl, Or selected from mono and dialkylaminoalkyl, and carboxyalkyl, or R 'and R "may be joined to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, and methylene adjacent to nitrogen is amino Optionally substituted with alkyl, carboxy or carboxyalkyl groups) and O-amino acids comprising O-CO-NH- (CH 2 ) m -CH- (NH 2 ) COOH, wherein m is 1-4. Selected from the group consisting of
    [46] R 6 and R 7 may be H, or may be joined to form a double bond,
    [47] R 8 is selected from H, halogen, for example F or Cl, alkyl, amino, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl and CF 3 . Furthermore, when R 1 to R 5 and R 9 to R 12 are OH and are present on adjacent ring carbons, these are methylene (—O—CH 2 —O—) or carbonyl (—O—CO—O—) groups. Can be joined to form a cyclic ring. Most preferred are 6,7 and 7,8-methylenedeoxy and 3 ', 4'-carbonyloxy (cyclic carbonate) derivatives.
    [48] Such methods can be performed for the purpose of treating any disease or disorder characterized by excessive or aberrant activity of T-lymphocytes. Such diseases and disorders include autoimmune disorders, diabetes mellitus, amyotrophic lateral sclerosis (ALS), rheumatoid arthritis, systemic lupus erythematosus (SLE), transplant host disease, transplant rejection, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, Behcet's disease, bullous swelling, cardiomyopathy, abdominal sprue dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, wife Leug-Strauss syndrome, scar swelling, CREST syndrome, cold coagulation disease, Crohn's disease, discoid lupus, essential mixed cold globulinemia, fibromyalgia-fibromyalitis, Graves disease, Guillain-Barré, Hashimoto thyroiditis, idiopathic lung Fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, insulin dependent diabetes mellitus, juvenile arthritis, lichen planus, meniere iere) disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, vulgaris ulcer, pernicious anemia, nodular polyarteritis, multiple chondritis, polysecretory syndrome, rheumatoid polymyalgia, multiple myositis-dermal myositis, primary agammaglobulinemia , Primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjogren's syndrome (primary or secondary), Stiff-Man syndrome, Dakayas arteritis, temporal arteritis / Giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, Wegener's granulomatosis, as well as the various inflammations and other diseases and disorders shown in the following detailed description of the present invention.
    [49] In addition, according to the present invention, when the method is carried out to treat diabetes or to stabilize blood glucose levels in a patient, the compound is luteolin (ie, X = O and R 2 , R 3 , R 9 , R 11 = OH Phosphorus 3 ', 4', 5,7-tetrahydroxyflavone compound), 5 glucoside of luteolin, 7 glucoside of luteolin, or any compound of formula I except apigenin .
    [50] Furthermore, according to the present invention, when the method is carried out for the treatment or prevention of amyotrophic lateral sclerosis (ALS), the method may comprise luteolin alone, genistein alone or dyedzein alone, or luteolin, genistein and Administration of one or more compounds of formula (I), except for possible combinations of two or more compounds selected from the group consisting of daidzein.
    [51] According to the present invention, there is also provided a method of inhibiting ion flux through a particular intracellular ion channel, such as a Kv1.3 channel, by administering an effective amount of the compound of formula (I). Among these compounds are substituted 3-deoxyflavonoids, for example. These compounds of formula (I) may be prepared by other agents such as other IKCa1 blockers such as clotrimazole, 1-[(2-chlorophenyl) diphenylmethyl] -1H-pyrazole, 2-chlorophenyl diphenyl cyano Methane or cyclosporin A or TNF-α inhibitors or other T-lymphocyte inhibitors, such as leflunomide, metabolites A-771726 [N- (4-trifluoromethylphenyl) -1-cyano-2- Ketopropyl-carboxamide] or N- [4- (trifluoromethyl) phenyl] -2-hydroxy-6-oxocyclopentanecarboxamide or N- [4- (trifluoromethyl) phenyl]- A method for immunosuppressive treatment of autoimmune disorders is provided, which may be administered in combination with 2-hydroxybenzamide and by administering a therapeutically effective amount of such a compound to a mammalian patient. Through inhibition of Kv1.3 channels, compounds of formula I can lower and stabilize blood glucose levels in type I and type II diabetes, and also improve symptoms of lupus erythematosus, autoimmune diseases affecting multiple organs. have. Type I diabetes is an autoimmune disease in which at least some patients autoreactive lymphocytes destroy β-cells in the pancreas, whereas the cause of type II diabetes is due to inadequate insulin production and reduced ability of insulin to affect target end organs. will be. Since ion channels play an important role in β-cell signaling in lymphocytes and pancreas, it is most likely that at least some of the therapeutic effects of flavonoids can occur through the blocking of ion channels. Using patch-clamp experiments, we found that Kv1.3, a voltage gate K + channel that regulates mitogenesis in human T lymphocytes, can be blocked by the compound of formula (I). The nontoxic nature of these flavonoids in humans makes them interesting as functional foods that can complement or replace existing medical or therapeutic therapies.
    [52] In addition, according to the invention, the method may comprise administering a compound according to formula (I) in combination with a vanadium element or vanadium containing compound (s). Vanadium compounds are being investigated clinically for their insulin mimic effects. Micromolar concentrations of vanadate and peroxovanadium compounds stimulate hexose in vivo and ex vivo, glucose oxidation and adipogenesis. Clinical trials demonstrating that sodium metavanadate and vanadil sulfate improve insulin sensitivity and fasting blood glucose levels have suggested the use of these agents in adjuvant therapy in diabetes. Combinations of vanadium compounds and 3-deoxyflavonoids can interact with lymphocytes and islet cells in an additive or perhaps synergistic manner, can be associated with their proposed clinical use, and new approaches to the management of diabetes and autoimmune disorders Get
    [53] In addition, according to the invention, certain compounds of formula (I), such as luteolin, are known to undergo first-pass metabolism when administered orally and absorbed through the gastrointestinal mucosa. Thus, such compounds may be administered by parenteral routes such as sublingual, buccal, intranasal, transdermal and the like (ie, where the compound is substantially absorbed through other than the gastric and / or intestinal mucosa). Such parenteral administration of these compounds can increase the circulating blood levels of the compounds and / or delay the metabolism of the compounds to increase their efficacy.
    [54] In addition, according to the present invention, the efficacy and / or duration of action of at least some of luteolin and other compounds of formula (I) may include rutin, or a congener or derivative of rutin, such compound (s) of formula (I). ) In combination with The potential effect of rutin and / or its analogues or derivatives is to slow the metabolic inactivation of luteolin or other compounds of formula I due to the inhibition of one or more liver enzymes (eg, certain cytochrome P450 enzymes). Can be.
    [55] Also according to the invention, a) luteolin, 5 (ie R 9 ) -glucoside of luteolin, 7 (ie R 11 ) -glucoside and apigenin of luteolin (ie X = O and R A method of treating diabetes or stabilizing blood glucose levels is provided by administering an effective amount of a compound of Formula (I), other than 4 ', 5,7-trihydroxyflavone compound of Formula (I), wherein 3 , R 9 , R 11 are OH.
    [56] Also according to the invention, luteolin (ie, 3 ′, 4 ′, 5,7-tetrahydroxyflavone compound of formula I, wherein X═O and R 2 , R 3 , R 9 , R 11 are OH), Genistein (i.e., 5,7-dihydroxy-3- (4-hydroxyphenyl) -4H-1benzopyran-4-one or 4 ', 5,7-trihydroxyisoflavone) or dydzein A therapeutically effective amount of a compound of formula I other than (7-hydroxy-3- (4-hydroxyphenyl) -4H-1benzopyran-4-one or 4 ', 7-dihydroxyisoflavone) Provided is a method of treating amyotrophic lateral sclerosis (ALS) in a patient by administering to the patient.
    [57] According to the present invention, by administering to a mammalian patient a therapeutically effective amount of at least one compound of formula I, cytokine secretion is inhibited and / or diabetes, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis and interstitial cystitis Provided are methods for treating an autoimmune disorder comprising.
    [58] In addition, according to the present invention, novel compositions of matter of formula II are provided.
    [59]
    [60] Wherein X is selected from O and S,
    [61] i) if X is O,
    [62] R 1 , R 4 , R 5 and R 8 are H or F,
    [63] R 6 and R 7 combine to form a double bond,
    [64] R 2 and R 3 are H, OH, SH, halogen, for example F or Cl, alkyl, amino, NHMe, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucolonide, and O-CO-A- (CH 2 ) n -NR 'R', wherein A is or is phenyl or substituted phenyl, n is 0 to 5, and R 'and R "are H, lower alkyl, hydroxyalkyl, aminoalkyl, mono and dialkylaminoalkyl, and Or R ′ and R ″ may be joined to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, and methylene adjacent to nitrogen is optionally substituted with an aminoalkyl, carboxy or carboxyalkyl group May be substituted) and O-amino acids including O-CO-NH- (CH 2 ) m -CH- (NH 2 ) COOH, wherein m is 1 to 4, and R 2 And when R 3 is OH or amino , They may be optionally bonded via methylene or carbonyl group,
    [65] R 9 is selected from OH, amino, NHMe, SH and SMe,
    [66] R 10 and R 11 or R 11 and R 12 are methylenedioxy (O-CH 2 -O) or cyclic carbonate (O-CO-O), or R 12 is H and R 10 , R 11 is H , OH, halogen, for example F or Cl, alkyl, amino, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucolonide, and O-CO-A- (CH 2 ) n -NR'R " where A is phenyl or substituted N is 0-5, and R 'and R "are selected from H, lower alkyl, hydroxyalkyl, aminoalkyl, mono and dialkylaminoalkyl, and carboxyalkyl, or R' and R" May combine to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, methylene adjacent to nitrogen may be optionally substituted with aminoalkyl, carboxy or carboxyalkyl groups) and O-CO-NH -(CH 2 ) m -CH- (NH 2 ) COOH, wherein m is from 1 to 4), provided that when R 2 and / or R 3 is H, OH, OMe, Cl or amino, R 9 , R 10 and R 11 are not the same;
    [67] ii) when X is S,
    [68] R 1 to R 5 and R 9 to R 12 are H, OH, halogen such as F or Cl, SH, SMe, alkyl, amino, NHMe, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycar Bonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucoronide, and O-CO-A — (CH 2 ) n —NR′R ″ wherein A is phenyl or substituted phenyl, or n is 0 to 5, R ′ and R ″ are H, lower alkyl, hydroxyalkyl, aminoalkyl, Mono or dialkylaminoalkyl, and carboxyalkyl, or R 'and R "combine to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, and methylene adjacent to nitrogen is aminoalkyl, Optionally substituted with carboxy or carboxyalkyl groups) and O-amino acids comprising O-CO-NH- (CH 2 ) m -CH- (NH 2 ) COOH, wherein m is 1 to 4 Selected from
    [69] R 6 and R 7 combine to form a double bond,
    [70] R 8 is selected from H and F,
    [71] When R 1 to R 5 and R 9 to R 12 are OH, SH or amino and are present on adjacent ring carbons, they are methylene (-O-CH 2 -O-) or carbonyl (-O-CO-O- , O-CO-NH- or -S-CO-NH-) can be bonded to form a cyclic ring.
    [72] In addition, according to the present invention, synthetic reactions and procedures which can be used to prepare the compounds of the present invention (Formulas (I) and (II)) are known in the art and can be applied by those skilled in the art. By way of example, hydroxy substituted flavones can be treated with haloacetyl halides, and the haloalkyl ester product can then be treated with amines to obtain the O-aminoacyl derivatives of interest. Similarly, catechol derivatives can be treated with diethyl carbonate or ethylene carbonate in the presence of a base to insert -O-CO-O- functional groups.
    [3] The present invention relates generally to chemical compositions, formulations and methods for medical treatment, and more particularly to the use of certain substituted 3-deoxyflavonoid compounds for immunosuppressive treatment of autoimmune disorders or inflammatory diseases in mammalian patients. It is about.
    [73] The following detailed description and examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.
    [74] The present invention optionally comprises a Kv1.3 channel inhibitor, for example a substituted 3-deoxyflavonoid compound, optionally an IKCa1 blocker, for example clotrimazole, 1-[(2-chlorophenyl) diphenylmethyl] -1H-pyrazole, 2-chlorophenyl diphenyl cyanomethane or cyclosporin A or TNF-α inhibitors or other T-lymphocyte inhibitors such as leflunomide, metabolite A-771726 [N- (4 -Trifluoromethylphenyl) -1-cyano-2-ketopropyl-carboxamide] or N- [4- (trifluoromethyl) phenyl] -2-hydroxy-6-oxocyclopentanecarboxamide or Provided is a pharmaceutical formulation comprising N- [4- (trifluoromethyl) phenyl] -2-hydroxybenzamide and provided by a therapeutically effective amount of such a compound in a mammalian patient for the treatment of immunosuppression of an autoimmune disorder do.
    [75] Moreover, the present invention deals with ion channels that constitute a set of molecular targets where these 3-deoxyflavonoids exert their biological effects. The flavonoid compounds of the present invention can lower and stabilize blood sugar levels in type I and type II diabetes and also improve symptoms of lupus erythematosus, autoimmune diseases affecting multiple organs. Type I diabetes is an autoimmune disease in which autoreactive lymphocytes destroy β-cells in the pancreas, while the cause of type II diabetes is due to inadequate insulin production and reduced ability of insulin to affect target end organs. Since ion channels play an important role in β-cell signaling in lymphocytes and pancreas, it is most likely that at least some of the therapeutic effects of flavonoids can occur through the blocking of ion channels. Using patch-clamp experiments, we surprisingly found that Kv1.3, a voltage gate K + channel that regulates mitotic reproduction in human T lymphocytes, can be blocked by these 3-deoxy flavonoids (Formula I). Found. The nontoxic nature of these flavonoids in humans makes them interesting as functional foods that can complement or replace existing medical or therapeutic therapies.
    [76] The present invention relates to a method of inhibiting the immune system in a subject in need thereof. Specifically, the method of the present invention is an autoimmune disease such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, ALS, multiple sclerosis, myasthenia gravis, type I and II diabetes mellitus, nephrotic syndrome, steroid dependence and steroid resistance Nephropathy, palmar-plantar pustules, allergic encephalomyelitis, glomerulonephritis, Behcet's syndrome, ankylosing spondylitis, multiple myositis, fibromyalitis and the like.
    [77] According to the present invention, by administering to a mammalian patient a therapeutically effective amount of at least one compound of formula I, T-lymphocyte activity is inhibited (by thereby inhibiting cytokine secretion and / or diabetes, ALS, multiple sclerosis, systemic redness) Methods of treating autoimmune disorders or any other diseases or disorders described in the Summary of the Invention, including seminal lupus, rheumatoid arthritis and interstitial cystitis. The following compounds of formula I are currently preferred for use in the methods of the present invention:
    [78] 6,7 methylenedioxy-3 ', 4', 5-trihydroxyflavone
    [79] 7,8 methylenedioxy-3 ', 4', 5-trihydroxyflavone
    [80] 6,7-carbonyloxy-3 ', 4', 5-trihydroxyflavone
    [81] 3 ', 4'-carbonyloxy-5,7-dihydroxyflavone
    [82] 3 ', 5,7-trihydroxyflavone-4'-phosphate
    [83] 3 ', 5', 7-trihydroxy-4 '-(2-amino-1-carboxypropyloxy) flavone
    [84] 5-hydroxy-3 ', 4', 7-tricarboxymethyloxyflavone
    [85] In addition, isoflavonoids in which the phenyl substituent in the formula (I) occupy the 3 position are included in the present invention. In particular, the composition may comprise one or more isoflavones and more specifically genistein (ie, 5,7-dihydroxy-3- (4-hydroxyphenyl) -4H-1benzopyran-4 in addition to one or more 3-deoxyflavonoids). -One or 4 ', 5,7-trihydroxyisoflavones) and / or didzein (ie, 7-hydroxy-3- (4-hydroxyphenyl) -4H-1benzopyran-4-one Or 4 ', 7-dihydroxyisoflavones).
    [86] When used to treat diabetes, the compound of formula I is luteolin (i.e. when R 2 , R 3 , R 9 , R 11 is OH (3 ', 4', 5,7-tetrahydroxyflavones) ) Or 5- (R 9 ) glucoside of luteolin, 7- (R 11 ) glucoside or apigenin of luteolin (ie, when R 3 , R 9 , R 11 is OH (4 ′, 5,7) -Trihydroxyflavone)).
    [87] When used to treat amyotrophic lateral sclerosis (ALS), it is preferred that the compound of formula (I) is neither luteolin alone nor genistein alone, nor is it a died agent alone.
    [88] In addition, the compounds of the present invention can be used in the skin findings of inflammatory, proliferative and hyperproliferative skin diseases and immune mediated diseases (eg, psoriasis, psoriatic arthritis, atopic dermatitis, contact dermatitis and other eczema dermatitis, seborrheic dermatitis). , Squamous gland, swelling swelling, bullous swelling, bullous epidermal detachment, angioedema, vasculitis, erythema, cutaneous eosinophilia, acne, alopecia areata and atherosclerosis).
    [89] In addition, the compounds of the present invention include respiratory diseases such as sarcoidosis, pulmonary fibrosis, idiopathic interstitial pneumonia and reversible obstructive airway diseases (asthma including bronchial asthma, allergic asthma, endogenous asthma, exogenous asthma and dust asthma, and bronchitis, etc.). More useful for treating the same symptoms). This compound may also be useful for treating liver damage associated with ischemia.
    [90] In addition, the compounds of the present invention may be useful in certain ocular diseases (e.g., keratoconjunctivitis, spring conjunctivitis, keratitis, uveitis, corneal white plaque, swelling, Mooren's ulcer, scleritis, Graves' ophtalmopathy and glia) It may be used for the treatment of emotional ophthalmitis.
    [91] The compounds also include inflammatory bowel disease (e.g. Crohn's disease), neurological diseases (including Guillain-Barré syndrome, Meniere name, neuromyopathy), endocrine diseases (including hyperthyroidism and Basedow's disease), Blood diseases (pure erythrocytosis, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis and lack of erythropoiesis), bone diseases (including osteoporosis), respiratory diseases (including sarcoidosis, idiopathic epileptic pneumonia) , Skin diseases (skin myositis, common white dermatosis, common scales, photoallergic-sensitive skin T-cell lymphoma), genital diseases (testitis, vulvitis), circulatory diseases (atherosclerosis, nodular polyarteritis, vasculitis, Börger's disease) (Buerger's disease and cardiomyopathy), collagen diseases (skin sclerosis, aortic syndrome, eosinophilic fasciitis, Wegener's granul) omatosis), including Sjogren's syndrome, periodontal disease), kidney disease (including kidney syndrome, hemolytic uremic syndrome, Goodpasture's syndrome) and muscular dystrophy. This compound is associated with enteritis / allergy (e.g., intraperitoneal disease, proctitis, ulcerative colitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis), and food-related allergies with signs that are not related to the gastrointestinal tract. It is also useful for treating diseases including sexual disorders (eg migraine, rhinitis and eczema). The invention can also be used for the prevention or treatment of vascular damage, ischemic bowel disease caused by inflammation of mucous membranes or blood vessels (eg, leukotriene mediated diseases), gastric ulcers, ischemic diseases and thrombosis. In addition, the present invention will be useful for treating tumor cells' resistance to multidrugs (ie, increasing the activity and / or sensitivity of chemotherapeutic agents).
    [92] Compounds of the present invention may be used for immunogenic diseases (e.g., chronic autoimmune liver diseases including autoimmune hepatitis, primary biliary sclerosis and sclerotic cholangitis), partial liver resection, acute liver necrosis (toxin, viral hepatitis, shock or oxygen free) Necrosis caused by the disease), hepatitis B-viral hepatitis B, non-A / non-B hepatitis, and sclerosis can also be useful in treating and preventing liver diseases.
    [93] Anecdotal studies in volunteers with type I and type II diabetes have shown that 3-deoxyflavonoids of formula (I) lower the amount of insulin needed for diabetics and have additional beneficial effects in the treatment of other autoimmune diseases. Based on the blocking of ion channels). The results of these ongoing studies show that when orally administered 3-deoxyflavonoids of Formula I in the form of powders or fast-acting tablets, elevated refractory blood glucose levels and glycosylated hemoglobin (Hb A1C ) levels are markedly reduced, Even small amounts of insulin normalize blood glucose levels in type I diabetics and show that other symptoms, such as peripheral neuropathy in diabetic patients with worsening symptoms, are reduced. It is believed that 3-deoxyflavonoids are absorbed in vivo through the buccal and sublingual mucosa. The volunteer's anecdotal study results also suggest 3-deoxyflavonoid efficacy in lupus erythematosus and multiple sclerosis. In addition, the inventors have found that inhibiting lymphocytes or pancreatic potassium channels includes various biological activities reported for 3-deoxyflavonoids of formula (I), anti-inflammatory effect, anti-inflammatory effect, LDL cholesterol lowering effect, antimutagenic effect and anticancer effect. Claim to contribute to Certain compounds included in Formula I are safe, naturally occurring flavones, which may be used as functional foods.
    [94] By targeting specific channels or combinations of channels, it is a major concern that both primary and secondary immune responses can be inhibited by non-toxic small molecules. It is also contemplated that blocking Kv1.3 channels alone or in combination with IKCa1 may provide a much more effective treatment for multiple sclerosis and other autoimmune diseases.
    [95] While the invention has been described in connection with the specific preferred embodiments in the above text and numerous details have been set forth in order to illustrate the invention, additional embodiments of the invention are possible and within the details set forth herein. It will be apparent to those skilled in the art that some of the variations can be varied in many ways without departing from the basic principles of the invention.
    [96] The following specific examples are set forth in order to illustrate the invention in detail and should not be considered as limiting the invention in any way.
    [97] Example 1
    [98] Fanger, J. Biol. Chem. 276: 12249, Kv1.3 channel suppression was performed using the patch clamp method described. As shown in FIG. 1, it was found that luteolin blocks approximately 25% of the current in the Kv1.3 channel at 20 μM and 100% at 100 μM compared to the control. These results clearly show that luteolin and related 3-deoxyflavonoids are effective at blocking Kv1.3 channels and thus effective at treating autoimmune diseases.
    [99] Example 2
    [100] Effect of 3-deoxyflavonoids on T lymphocyte cytotoxicity
    [101] Luteolin was 264 peptide-pulsed using a cytotoxic T lymphocyte cell line (named CTL-CTL264) specific for the peptide antigen (referred to as aa 264-272, 264 peptide) derived from the tumor suppressor protein p53. The effect on CTL cytotoxicity against T2 target cells was tested. T2 target cells were pulsed with 264 peptide for 2 hours, labeled with calcein-AM for 30 minutes, and washed three times. The mixture of T2 cells and CTL264 was incubated for 4 hours. 100 μl of supernatant was transferred to a 96-well flat-bottom microtiter plate and fluorescence (538 nm) was read to determine the amount of calcein released due to cell lysis. Surprisingly, we found that luteolin significantly inhibits CTL cytotoxicity against 264-pulsed T2 target cells in a concentration dependent manner (FIG. 2).
    [102] Example 3
    [103] The amount of calcein released spontaneously from cytotoxic T lymphocytes was measured by culturing target cells in RPMI-10. Maximum release of calcein was determined by culturing target cells in Triton X-10. Data was recorded as the average value of the values measured repeatedly three times. 3 shows inhibition of calcein release by 3-deoxyflavonoids luteolin and tricetin.
    [104] Example 4
    [105] The effect of luteolin treatment on rats with chronic type I (BBWor) diabetes was investigated. In this study, lean male rats with diabetes were randomly assigned to three treatment groups (3-4 rats / group). (1) gastrointestinal administration of 3 mg of luteolin to each group; (2) subcutaneous injection (0.9-1.2 mU / day) of PZI insulin; Or (3) no treatment. Blood glucose levels were measured from 0 hours to 6 hours. Data is expressed as mean blood glucose over time after luteolin treatment.
    [106] Rats injected with insulin once had a 75% decrease in blood glucose levels (reduction from 415 mg / dl to 112 mg / dl) within 6 hours after injection. This response was in full agreement with the results of our previous studies in rat models with type 1 (BBWor) diabetes. More notably, administration of luteolin to diabetic rats resulted in a 31% decrease in blood glucose levels at 6 hours (from 445 mg / dl / to 307 mg / dl). In comparison, the control group did not reduce hyperglycemic symptoms during the same time interval (change from 414 mg / dl to 404 mg / dl). Thus, a single dose of 3 mg of luteolin in rats with insulin-dependent diabetes mellitus (Type I BBWor) could reduce hyperglycemic symptoms by 31% within 6 hours.
    [107] Example 5
    [108] In addition, the effect of luteolin on reducing hyperglycemic symptoms in rats with chronic type 2 diabetes was evaluated. In this study, luteolin dose and treatment frequency were increased to compensate for the increased metabolism of obese rats. Initially, basic studies were performed for 24 hours on 9 rats with chronic type 2 diabetes. As a result of analyzing the rats with diabetes for 24 hours, there was no significant change in symptoms of hyperglycemia. These same rats were randomly assigned to three groups and they were administered 50 mg, 150 mg and 250 mg of luteolin over three times (11AM, 2PM and 8PM) for 24 hours. Blood glucose analysis was measured every 2 hours.
    [109] 4 shows the effect of luteolin administration in rats with type II diabetes. Rats that received 50 mg (minimum) of luteolin three times daily (150 mg total) had a 10.2% decrease in blood glucose levels within 24 hours of treatment. In comparison, rats receiving 150 mg (intermediate dose) of luteolin (450 mg total) had a 22.9% decrease in blood glucose levels. Rats of the third experimental group administered luteolin at 250 mg (maximum amount) (total 750 mg) showed a decrease in maximal blood glucose levels of 27.7%. Interestingly, a 52% reduction in blood glucose levels was observed (18 mg from 777 mg / dl to 372 mg / dl) within 18 hours of treatment in one of the rats administered medium doses of luteolin. Unfortunately, the animal accidentally penetrated the esophagus during drug administration and died at some time before the 24 hour time point. These results demonstrated that administration of luteolin to rats with type 2 diabetes significantly reduced hyperglycemia 10-28% at 24 hours, and that these observations depended on the dose of luteolin.
    [110] Example 6
    [111] In this example, the luteolin dose was normalized to 50 mg of luteolin three times daily in the same group of type II BBZDR rats of Example 5 over an extended treatment period of two weeks. Changes in this method resulted in further decreases in blood glucose levels. Data is expressed as percent change in blood glucose level% for each individual pretreatment level for each rat.
    [112] In FIG. 5, administration of 50 mg of luteolin to rats with obesity type II (BBZDR) diabetes three times a day for two weeks reduced blood glucose levels in almost all rats (excluding rat 3, range of reduction: 36 to 36). 54%). Rat 3, in which the esophageal canal was found at necropsy, had a 9.3% increase in blood glucose, which probably inhibited the response to effective administration and treatment. Overall, administration of luteolin to type 2 diabetic rats resulted in an average 41.1% reduction in blood glucose levels (from 660 mg / dl to 389 mg / dl).
    [113] Example 7
    [114] A 7 year old boy with antibodies to type I diabetes continued to receive prophylactic insulin (NPH 2 units, every night) since the child turned 2 years old. At seven years and four months of insulin, the child showed signs of blood glucose levels of 260 mg / dl, starting with four units of NPH each night to control blood glucose levels. Approximately 45 days after starting insulin treatment, patients were sublingually administered 75-150 mg of luteolin powder (75% luteolin, 25% routine) instead of insulin four times a day. There was no need to administer insulin to maintain an average blood glucose level of approximately 105 mg / dl. Approximately 20 days after administration of luteolin powder (75% luteolin, 25% routine), the dose of luteolin powder (75% luteolin, 25% routine) is gradually increased while continuously adjusting blood sugar and improving glucose tolerance. It was reduced to 25 mg by 4 times a day. Patient blood was collected and analyzed for comprehensive metabolic enzymes including liver function, lipids, glycosylated hemoglobin (Hb A1c ). All enzyme systems were normal and HbA1c was 8.0. After 90 days of 25 mg of luteolin powder (75% luteolin, 25% routine) four times daily, no blood glucose levels of 125 mg / dl or more were observed in the child, and the average blood glucose level of this child was 95 mg. / dl and Hb A1c was 6.3. Luteolin powder (75% luteolin, 25% routine) treatment and blood glucose concentrations are shown in FIG. 6.
    [115] Example 8
    [116] A 69-year-old white man with type II diabetes received Glusotrol XL 10 mg twice daily, Actos 45 mg once daily, Glucophage 1000 mg daily 2 Dosing was done twice. His average blood glucose over the year was approximately 170 mg / dl. His blood glucose data for one year prior to dosing is shown in FIG. 7. This patient was administered three times a day the composition described in Example 12. Patient blood glucose was measured periodically as shown in FIG. 8. The data shows a consistent decrease in blood glucose over approximately 60 days (not achieved using prescription drugs currently used in the art).
    [117] Example 9
    [118] A 47-year-old Caucasian woman weighing 65.8 kg with a 15-year history of multiple sclerosis was an acute onset of severe pain in the eyes and limbs. The patient was given beta interferon as directed by her doctor. Patients were sublingually administered 100 mg of luteolin powder (45% luteolin, 55% routine) four times daily. Within a short period of time, subjects showed changes in neuropathy symptoms consistent with rapid sublingual and buccal absorption. Within 24 hours after dosing, her symptoms decreased approximately 50% and the disease symptom was eliminated within 72 hours as a result of continuous administration of luteolin powder (45% luteolin, 55% routine) four times a day for 14 days. When discontinuation of the luteolin powder (45% luteolin, 55% routine), the subject experienced a relapse of multiple sclerosis symptoms within 5 days. Again, this female patient was administered 250 mg of luteolin powder (45% luteolin, 55% routine) four times a day for a day and thereafter luteolin powder (45% luteolin, 55% routine) 100 Continuous doses of mg were observed to reduce MS symptoms that persisted for 30 days.
    [119] Example 10
    [120] The treatment of anti-inflammatory and analgesic treatments for 32-year-old white men who have suffered for 10 years from fibromyalitis induced by serious motor vehicle accidents has not been successful. The joint pain he suffered during physical labor surpassed his patience, and he often lay down and rested because he could not move without serious pain. As a result of administering 50 mg of luteolin powder (75% luteolin, 25% routine) three times a day to this subject, it was observed that his symptoms were markedly reduced within two weeks, allowing him to return to work. He no longer needed analgesics and no symptoms recurred.
    [121] Example 11
    [122] The following compositions represent conventional formulations for the alleviation of autoimmune diseases. For each disease this composition may vary and is not to be construed as fixed.
    [123] β-carotene 6000 IU
    [124] Retinyl Palmitate 6000 IU
    [125] Ascorbic acid 375 mg
    [126] Ascorbyl Palmitate 25 mg
    [127] Cholecalciferol 400 IU
    [128] TPGS 500 mg
    [129] Phytonadione 150 mcg
    [130] Thiamine 15 mg
    [131] Riboflavin 15 mg
    [132] Niacin 15 mg
    [133] Niacinamide 1450 mg
    [134] Pyridoxine HCl 22.5 mg
    [135] Pyridoxal-5-phosphate 2.5 mg
    [136] Folic acid 800 mcg
    [137] Cyanocobalamin 60 mcg
    [138] Biotin 3 mg
    [139] Pantothenic Acid 100 mg
    [140] Calcium 200 mg
    [141] Iodine (KI) 150 mcg
    [142] Magnesium 500 mg
    [143] 15 mg of zinc
    [144] Selenium 300 mcg
    [145] 2 mg of copper
    [146] Manganese 5 mg
    [147] Chromium picolinate 400 mcg
    [148] Molybdenum 100 mcg
    [149] Boron 3 mg
    [150] Silica 10 mg
    [151] Vanadil sulfate 5 mg
    [152] Choline Bitartrate 50 mg
    [153] Concentrated Citrus Bioflavonoids 500 mg
    [154] (Citrus Bioflavonoid Conc.)
    [155] Lipoic acid 50 mg
    [156] Lutein 3 mg
    [157] 3-deoxyflavonoids 25 mg
    [158] Lycopene 2 mg
    [159] N-acetyl cysteine 200 mg
    [160] Taurine 500 mg
    [161] 4-carboxy-2-thiazolidone 100 mg
    [162] PEG-400 25 mg
    [163] Example 12
    [164] Tablet compositions for oral administration were prepared containing the following ingredients.
    [165] Vitamin C (ascorbic acid and calcium ascorbate) 65 mg
    [166] Vitamin D (cholecalcinphenol) 50 IU
    [167] Vitamin E (δ-α Tocopheryl Acetate) 5 IU
    [168] Niacin (niacinamide) 10 mg
    [169] Biotin 100 mcg
    [170] Calcium (Calcium Carbonate) 160 mg
    [171] Chromium (Chromium Polynicotinate) 100 mcg
    [172] Vanadium (Vanadil Sulfate) 10 mcg
    [173] 3-deoxyflavonoids 25 mg
    [174] (50% luteolin, 50% routine)
    [175] Microcrystalline Cellulose 35 mg
    [176] Croscarmellose Sodium 7 mg
    [177] Stearic acid 10.5 mg
    [178] Magnesium stearate 3.2 mg
    [179] Silicon dioxide 1.8 mg
    [180] Opadry NS Y-40-19133 3.5 mg
    [181] Although the present invention has been described with reference to specific embodiments, it should be understood by those skilled in the art that various changes may be made in the present invention and equivalents may be substituted without departing from the scope of the present invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, method and / or step or steps of the method to fall within the scope of the invention. Accordingly, the scope of the invention should be determined with reference to the appended claims, along with the full scope of equivalents possessed by such claims.
    权利要求:
    Claims (37)
    [1" claim-type="Currently amended] A compound of formula (I)
    <Formula I>

    Wherein X is selected from O and S,
    i) if X is O,
    R 1 , R 4 , R 5 and R 8 are H or F,
    R 6 and R 7 combine to form a double bond,
    R 2 and R 3 are H, OH, SH, halogen, alkyl, amino, HNMe, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucolonide, and O-CO-A- (CH 2 ) n -NR'R " where A Is phenyl or substituted phenyl or is absent, n is 0 to 5 and R 'and R "are selected from H, lower alkyl, hydroxyalkyl, aminoalkyl, mono and dialkylaminoalkyl, and carboxyalkyl, or R 'And R' may combine to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, and methylene adjacent to nitrogen may be optionally substituted with aminoalkyl, carboxy or carboxyalkyl groups) and O -CO-NH- (CH 2 ) m is selected from the group consisting of O-amino acids comprising -CH- (NH 2 ) COOH, wherein m is 1 to 4, wherein R 2 and R 3 are OH, SH Or amino, these are methylene Or may be optionally bonded via a carbonyl group,
    R 9 is selected from OH, amino, NHMe, SH and SMe,
    R 10 and R 11 or R 11 and R 12 are methylenedioxy (O-CH 2 -O) or cyclic carbonate (O-CO-O), or R 12 is H and R 10 , R 11 is H , OH, halogen, for example F or Cl, alkyl, amino, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucolonide, and O-CO-A- (CH 2 ) n -NR'R " where A is phenyl or substituted N is 0-5, and R 'and R "are selected from H, lower alkyl, hydroxyalkyl, aminoalkyl, mono and dialkylaminoalkyl, and carboxyalkyl, or R' and R" May combine to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, methylene adjacent to nitrogen may be optionally substituted with aminoalkyl, carboxy or carboxyalkyl groups) and O-CO-NH -(CH 2 ) m -CH- (NH 2 ) COOH, wherein m is from 1 to 4), provided that when R 2 and / or R 3 is H, OH, OMe, Cl or amino, R 9 , R 10 and R 11 are not the same;
    ii) when X is S,
    R 1 to R 5 and R 9 to R 12 are H, OH, halogen such as F or Cl, SH, SMe, alkyl, amino, NHMe, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycar Bonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucoronide, and O-CO-A — (CH 2 ) n —NR′R ″ wherein A is phenyl or substituted phenyl, or n is 0 to 5, R ′ and R ″ are H, lower alkyl, hydroxyalkyl, aminoalkyl, Mono or dialkylaminoalkyl, and carboxyalkyl, or R 'and R "combine to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, and methylene adjacent to nitrogen is aminoalkyl, Optionally substituted with carboxy or carboxyalkyl groups) and O-amino acids comprising O-CO-NH- (CH 2 ) m -CH- (NH 2 ) COOH, wherein m is 1 to 4 Selected from
    R 6 and R 7 combine to form a double bond,
    R 8 is selected from H and F,
    When R 1 to R 5 and R 9 to R 12 are OH and / or amino and are present on adjacent ring carbons, these are methylene (-O-CH 2 -O-) or carbonyl (-O-CO-O -, -O-CO-NH- or -S-CO-NH-) can be bonded to form a cyclic ring.
    [2" claim-type="Currently amended] The compound of claim 1, wherein R 10 and R 12 are OH.
    [3" claim-type="Currently amended] The compound of claim 1, which is 5-hydroxy-3 ', 4', 7-tricarboxymethyloxyflavone.
    [4" claim-type="Currently amended] The compound of claim 1, which is 6,7-methylenedioxy-3 ', 4', 5-trihydroxyflavone.
    [5" claim-type="Currently amended] The compound of claim 1, which is 7,8-methylenedioxy-3 ', 4', 5-trihydroxyflavone.
    [6" claim-type="Currently amended] The compound of claim 1, which is 6,7-carbonyloxy-3 ′, 4 ′, 5-trihydroxyflavone.
    [7" claim-type="Currently amended] The compound of claim 1, which is 3 ', 4'-carbonyloxy-5,7-dihydroxyflavone.
    [8" claim-type="Currently amended] The compound of claim 1, which is 3 ', 5,7-trihydroxyflavone-4'-phosphate.
    [9" claim-type="Currently amended] The compound of claim 1, which is 3 ', 5,7-trihydroxy-4'-(2-amino-1-carboxypropyloxy) flavone.
    [10" claim-type="Currently amended] A method of inhibiting T-lymphocyte activity in a patient comprising administering a compound of Formula I to a human or veterinary patient in an amount effective to inhibit T-lymphocyte activity.
    <Formula I>

    Wherein X is selected from O and S,
    R 1 to R 5 and R 9 to R 12 are H, OH, SH, Sme, halogen, alkyl, amino, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl, O-hydroxyalkyl, CF 3 , O-alkyl, O-SO 3 H, O-SO 2 H, O-PO 3 H, O-glycoside, O-glucolonide, and O-CO-A- (CH 2 ) n -NR ' R ″ (wherein A is phenyl or substituted phenyl or n is 0 to 5, R ′ and R ″ are H, lower alkyl, hydroxyalkyl, aminoalkyl, mono and dialkylaminoalkyl, and carboxy Or R ′ and R ″ may be joined to form a cyclic ring optionally substituted with O, S, NH or N-alkyl, and methylene adjacent to nitrogen is optionally substituted with an aminoalkyl, carboxy or carboxyalkyl group And O-amino acids, including O-CO-NH- (CH 2 ) m -CH- (NH 2 ) COOH, wherein m is 1 to 4, and
    R 6 and R 7 may be H, or may be joined to form a double bond,
    R 8 is selected from H, halogen, alkyl, amino, cyano, carboxyl, carboxyalkyl, carboxamide, alkoxycarbonyl and CF 3 , and R 1 to R 5 and R 9 to R 12 are OH, When SH or amino and present on adjacent ring carbons, these are methylene (-O-CH 2 -O-) or carbonyl (-O-CO-O-, -O-CO-NH- or -S-CO- May be joined via an NH—) group to form a cyclic ring. Most preferred are 6,7 and 7,8-methylenedeoxy and 3 ', 4'-carbonyloxy (cyclic carbonate) derivatives.
    [11" claim-type="Currently amended] The method of claim 10, wherein the method is performed to treat diabetes or to stabilize a patient's blood glucose level, wherein the compound is not luteolin, 5 glucosides of luteolin, 7 glucosides of luteolin and apigenin.
    [12" claim-type="Currently amended] The method of claim 10, wherein the method is performed to treat amyotrophic lateral sclerosis and wherein the compound is not luteolin, genistein and dyedzein.
    [13" claim-type="Currently amended] The method of claim 10, which is carried out to treat amyotrophic lateral sclerosis, comprising administering the compound of formula of claim 10 in combination with another compound.
    [14" claim-type="Currently amended] The method of claim 10, wherein the compound is administered in combination with Rutin, Rutin congener or Rutin Derivative.
    [15" claim-type="Currently amended] The method of claim 14, wherein a) the compound of claim 10 and b) the rutin, rutin, or rutin derivative are administered at a weight ratio of about 50% / 50%.
    [16" claim-type="Currently amended] The method of claim 14, wherein a) the compound of claim 10 and b) the rutin, rutin, or rutin derivative are administered at a weight ratio of about 75% / 25%.
    [17" claim-type="Currently amended] The method of claim 14, wherein a) the compound of claim 10 and b) the rutin, rutin, or rutin derivative are administered at a weight ratio of about 50% / 50% to about 75% / 25%.
    [18" claim-type="Currently amended] The method of claim 10, wherein the first pass metabolism occurs when the compound of claim 10 is absorbed through the gastric and / or intestinal mucosa, and the compound of claim 10 is substantially absorbed by a route other than the gastric and / or intestinal mucosa. How to administer.
    [19" claim-type="Currently amended] The method of claim 18, wherein the compound is administered for substantially absorption through the sublingual mucosa of the patient.
    [20" claim-type="Currently amended] The method of claim 18, wherein the compound is administered for substantially absorption through the buccal mucosa of the patient.
    [21" claim-type="Currently amended] The method of claim 18, wherein the compound is administered for substantially absorption through the rectal mucosa of the patient.
    [22" claim-type="Currently amended] The method of claim 18, wherein the compound is administered for substantially absorption through the nasal mucosa of the patient.
    [23" claim-type="Currently amended] The method of claim 18, wherein the compound is administered for substantially absorption through the sublingual mucosa of the patient.
    [24" claim-type="Currently amended] The method of claim 18, wherein the compound is administered for substantially absorption through the skin of the patient.
    [25" claim-type="Currently amended] The method of claim 18, wherein the compound is administered by injection.
    [26" claim-type="Currently amended] The method of claim 10, wherein R 10 and R 12 are OH.
    [27" claim-type="Currently amended] The method of claim 10, wherein the compound is 6,7-methylenedioxy-3 ', 4', 5-trihydroxyflavone.
    [28" claim-type="Currently amended] The method of claim 10, wherein the compound is 7,8-methylenedioxy-3 ', 4', 5-trihydroxyflavone.
    [29" claim-type="Currently amended] The method of claim 10, wherein the compound is 6,7-carbonyloxy-3 ′, 4 ′, 5-trihydroxyflavone.
    [30" claim-type="Currently amended] The method of claim 10, wherein the compound is 3 ', 4'-carbonyloxy-5,7-dihydroxyflavone.
    [31" claim-type="Currently amended] The method of claim 10, wherein the compound is 3 ', 5,7-trihydroxyflavone-4'-phosphate.
    [32" claim-type="Currently amended] The method of claim 10, wherein the compound is a 3 ′, 5,7-trihydroxy-4 ′-(2-amino-1-carboxypropyloxy) flavone.
    [33" claim-type="Currently amended] The method of claim 10, wherein the compound is 5-hydroxy-3 ', 4', 7-tricarboxymethyloxyflavone.
    [34" claim-type="Currently amended] The method of claim 10, wherein the compound is luteolin.
    [35" claim-type="Currently amended] The method of claim 10, wherein the compound is luteolin and further comprising administering to the patient a routine, routine equivalent, or routine analogue in an amount effective to enhance the efficacy or time of action of the luteolin.
    [36" claim-type="Currently amended] The compound of claim 10, wherein the compound is genistein (5,7-dihydroxy-3- (4-hydroxyphenyl) -4H-1benzopyran-4-one or 4 ', 5,7-trihydroxyiso Flavones).
    [37" claim-type="Currently amended] The compound of claim 10, wherein the compound is combined with a didzein (7-hydroxy-3- (4-hydroxyphenyl) -4H-1benzopyran-4-one or 4 ′, 7-dihydroxyisoflavone) How to administer.
    类似技术:
    公开号 | 公开日 | 专利标题
    Morley et al.2007|Tangeretin and nobiletin induce G1 cell cycle arrest but not apoptosis in human breast and colon cancer cells
    Bischoff2008|Quercetin: potentials in the prevention and therapy of disease
    Zhan et al.2006|Protective effects of isoliquiritigenin in transient middle cerebral artery occlusion-induced focal cerebral ischemia in rats
    CA2308530C|2004-09-28|Method for inhibiting or reducing the risk of macular degeneration
    Birt et al.2001|Dietary agents in cancer prevention: flavonoids and isoflavonoids
    Meyer et al.2006|Bioavailability of apigenin from apiin-rich parsley in humans
    Mamede et al.2011|The role of vitamins in cancer: a review
    EP1071419B1|2007-07-25|Cannabinoids as antioxidants and neuroprotectants
    JP5535999B2|2014-07-02|Use of equol to treat androgen-mediated diseases
    US7611738B2|2009-11-03|Processes for extracting phytochemicals from pomegranate solids and compositions and methods of use thereof
    Maher et al.2005|Flavonoids protect retinal ganglion cells from oxidative stress–induced death
    Lee et al.2007|Effect of flavonoids on human health: old subjects but new challenges
    Lakhanpal et al.2007|Quercetin: a versatile flavonoid
    US7943164B2|2011-05-17|Composition and method for treating diabetes and metabolic disorders
    US6579544B1|2003-06-17|Method for supplementing the diet
    US8591964B2|2013-11-26|Vaccinium species compositions
    US7714151B2|2010-05-11|Aminated isoflavonoid derivatives and uses thereof
    JP4205943B2|2009-01-07|Bioavailable compositions of natural and synthetic HCA
    López-Posadas et al.2008|Effect of flavonoids on rat splenocytes, a structure–activity relationship study
    CA2287965C|2009-10-20|Treatment or prevention of menopausal symptoms and osteoporosis
    US6426362B1|2002-07-30|Formulations of tocopherols and methods of making and using them
    Shia et al.2009|Metabolism and pharmacokinetics of 3, 3′, 4′, 7-tetrahydroxyflavone |, 5-hydroxyflavone, and 7-hydroxyflavone and antihemolysis effects of fisetin and its serum metabolites
    US6592910B1|2003-07-15|Methods of treating clinical diseases with isoflavones
    EP1589964B1|2011-11-23|Compounds having anti-proliferative properties
    Kim et al.2004|Chemoprevention by grape seed extract and genistein in carcinogen-induced mammary cancer in rats is diet dependent
    同族专利:
    公开号 | 公开日
    AU2002332878A1|2003-03-24|
    US6774142B2|2004-08-10|
    JP2005504796A|2005-02-17|
    MXPA04002077A|2005-02-17|
    WO2003022994A3|2003-10-09|
    BR0212354A|2004-11-30|
    WO2003022994A2|2003-03-20|
    EP1429750A4|2005-08-03|
    US20030069192A1|2003-04-10|
    DE60237213D1|2010-09-16|
    CN1668287A|2005-09-14|
    EP1429750B1|2010-08-04|
    AT476178T|2010-08-15|
    US20040209825A1|2004-10-21|
    EP1429750A2|2004-06-23|
    CN1668287B|2012-03-28|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    2001-09-06|Priority to US31766601P
    2001-09-06|Priority to US60/317,666
    2002-08-30|Priority to US40712502P
    2002-08-30|Priority to US60/407,125
    2002-09-06|Application filed by 시녹스, 인크.
    2002-09-06|Priority to PCT/US2002/028348
    2004-06-09|Publication of KR20040048407A
    优先权:
    申请号 | 申请日 | 专利标题
    US31766601P| true| 2001-09-06|2001-09-06|
    US60/317,666|2001-09-06|
    US40712502P| true| 2002-08-30|2002-08-30|
    US60/407,125|2002-08-30|
    PCT/US2002/028348|WO2003022994A2|2001-09-06|2002-09-06|Inhibition by 3-deoxyflavonoids of t-lymphocyte activation and therapies related thereto|
    [返回顶部]