JP5047431B2 - Glucosamine and eggs reduce inflammation - Google Patents

Description

【０１１７】
超免疫卵製品を得るための免疫処理手順
病原体死菌調製品を前記のように調製した。最初のワクチン処理のために、細菌を完全フロイントアジュバントと混合し、細菌材料５．６ｍｇをニワトリの胸部筋肉中に注射した。その後のワクチン処理のために、細菌調製品を不完全フロイントアジュバントと混合し、２週間隔で６か月間ニワトリに注射した。
【０１１８】
実施例２
目的
炎症の慢性動物モデルである確立されたラットアジュバント関節炎において、グルコサミン塩酸およびＤＣＶ超免疫卵の共同的抗関節炎作用を評価すること。
【０１１９】
結果
表２は、試験の１４日、２２日および３０日の足の浮腫に関する全データを示す。その結果から、ＰＬ−１００＋グルコサミンＨＣｌ群が、疾患の１４日および３０日に最高のパーセント炎症阻止率を示したことがわかる。ＰＬ−１００卵およびグルコサミンＨＣｌは、試験の１４日および２２日に単独で阻止作用を示したが、疾患が急速に進行して重症多発性関節炎となった３０日ではいずれも無効であった。
【０１２０】
表３は、試験の１６日、２３日および３０日の全ての血清フィブリノーゲン量を示す。その結果から、フィブリノーゲン量は始め１６日に増加した後、２３日および３０日に減少することがわかる。しかし、１６日の増加量は、ＰＬ−１００およびグルコサミンＨＣｌ単独と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最小であり、血清フィブリノーゲン量の阻止も、ＰＬ−１００およびグルコサミンＨＣｌ単独と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最高である。また、３０日では、ＰＬ−１００＋グルコサミンＨＣｌ群におけるフィブリノーゲン量のパーセント阻止率は、インドメタシン治療群の４６％に対し、３２％である。
【０１２１】
表４は、試験の２２日および３０日の全ての組織形態観察を示す。その結果から、（関節周囲の滑膜を含め、軟組織中への単核および／または多形核炎症細胞の浸潤を示す）関節周囲／軟組織の炎症は、２２日、３０日のいずれでも、他の群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最も影響が小さいことがわかる。
【０１２２】
（特に骨膜下区域において、新骨成長量の増加、過剰を示す）過骨症は、他の群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で２２日に最小で、３０日では小さい。
【０１２３】
２２日および３０日の軟骨侵食は、他の群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最小である。
【０１２４】
（関節表面での炎症細胞、浸出液、フィブリンまたは繊維組織の蓄積を示す）パンヌス軟骨は、他の群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で２２日および３０日に最も影響が小さい。
【０１２５】
（関節表面の間に繊維組織および炎症組織が形成され、普通軟骨が完全に消失し、この区域が繊維組織で置換される結果、接着および関節腔が生じるためにできる肥大し、運動性を失った関節を示す）強直症は、任意の他の群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最小である。
【０１２６】
最後に、（骨の壊死および溶解を示す）骨溶解は、ある程度の溶解が起こる他の群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ治療群では２２日に存在しない。
【０１２７】
【表２】

【０１２８】
【表３】

【０１２９】
【表４】

【０１３０】
まとめ
炎症の慢性動物モデルであるラットアジュバント関節炎モデルにおいて、ＰＬ−１００卵およびグルコサミンＨＣｌは、以下の点で付加的作用を示した。
ａ）動物の後部左足の腫脹を減少させ、それにより１４日、２２日および３０日での抗炎症作用を示した。
ｂ）注射した２２日の後、（炎症の認められた血清マーカーである）血清フィブリノーゲン量を減少させ、試験終了日の３０日までその状態を継続した。
ｃ）慢性関節リウマチの程度が頂点に達していた２２日に、単核および多形核細胞の浸潤による関節周囲／軟組織に対する作用を最小限に抑えた。
ｄ）試験の２２日に過骨症から完全に防御した。
ｅ）２２日に関節炎関節周囲の軟骨の侵食を完全に防御し、３０日に傷害を最小限に抑えた。
ｆ）２２日にパンヌス区域の軟骨を完全に防御し、３０日に傷害を最小限に抑えた。
ｇ）２２日に強直症から影響を受けた関節を完全に防御し、２２日に関節炎関節区域の骨を骨溶解から完全に防御した。
ｈ）図１は試験の１４日と２２日における足部浮腫の全データを示す。その結果は、ＰＬ−１００＋グルコサミンＨＣｌ群が疾患の１４日に最高のパーセント炎症阻止率を示したことを明らかにする。
ｉ）図２は試験の１６日、２３日および３０日における全ての血清フィブリノーゲン量を示す。その結果から、フィブリノーゲン量は始め１６日に増加した後、２３日および３０日に減少することがわかる。しかし、１６日の増加量は、ＰＬ−１００およびグルコサミンＨＣｌ単独と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最小であり、血清フィブリノーゲン量の阻止も、ＰＬ−１００およびグルコサミンＨＣｌ各単独群と比較して、ＰＬ−１００＋グルコサミンＨＣｌ群で最高である。また、３０日では、ＰＬ−１００＋グルコサミンＨＣｌ群におけるフィブリノーゲン量のパーセント阻止率は、インドメタシン治療群の４６％に対し、３２％である。したがって、ＰＬ−１００＋グルコサミンＨＣｌ群は、発症時に炎症の重度を抑制するだけでなく、（血清炎症マーカー量によって示されるように）試験の終了に向けて炎症を阻止する。
【図面の簡単な説明】
【図１】 ラットアジュバント関節炎モデルにおいて、超免疫卵およびグルコサミンの効果を示す棒グラフである。
【図２】 ラットアジュバント関節炎モデルにおいて、血漿フィブリノーゲン量を示す棒グラフである。[0001]
(Related application)
This application claims the benefits of US Provisional Patent Application No. 60 / 153,887 filed Sep. 14, 1999 and US Provisional Patent Application No. 60 / 192,386 filed Mar. 27, 2000. .
[0002]
(Background of the Invention)
Field of Invention
The present invention relates to a method for reducing inflammation in an animal. More particularly, the present invention relates to a combination of egg product and glucosamine to produce a synergistic effect in reducing inflammation in animals, particularly arthritis.
[0003]
inflammation
Inflammation as defined in the Dorland Medical Dictionary is a local protective reaction caused by tissue injury or destruction that destroys, dilutes, or sequesters both the cause of the injury and the damaged tissue. is there". Inflammation is characterized by a fenestration of the microvasculature, leakage of blood components into the interstitial space, and migration of leukocytes into the inflamed tissue. Macroscopically, inflammation is usually accompanied by commonly experienced clinical signs of erythema, edema, hyperalgesia (tenderness) and pain.
[0004]
During this complex reaction, chemical mediators such as histamine, 5-hydroxytryptamine, various chemotactic compositions, bradykinin, leukotrienes, prostaglandins are locally released. Phagocytic cells migrate into the area and the cell lysosomal membrane breaks, releasing lytic enzymes. All of these events can contribute to the inflammatory response.
[0005]
In the particular case of rheumatoid arthritis, the occurring inflammation appears to involve the binding of antigen and antibody complement, resulting in the local release of chemotactic and chemically activated compositions that attract leukocytes. Leukocytes phagocytose antigen-antibody and complement complexes and also release many enzymes contained in their lysosomes. These lysosomal enzymes then cause injury to cartilage and other tissues, which promotes the degree of inflammation. A cellular immune response may also be involved. Prostaglandins, which are important intracellular regulators of cell function, are also released during this process.
[0006]
An inflammatory response is any response characterized by inflammation as defined above. It is well known to those skilled in the medical art that the inflammatory response causes many physical discomforts (ie, pain and loss of function) that are associated with various diseases and injuries.
[0007]
arthritis
Arthritis appears in various forms. Some more common forms include rheumatoid arthritis, osteoarthritis and systemic rheumatism.
[0008]
Rheumatoid arthritis is an autoimmune disease characterized by joint pain, swelling and stiffness. Rheumatoid arthritis is a disease that affects about 3% of Americans, especially women. Rheumatoid arthritis is a highly disabling disease, with clinical disease occurring most in people aged 40-60 years but usually affecting adults aged 30 to 40 years. Although pharmacotherapy is effective to some extent, as many as 7% of patients with rheumatoid arthritis become somewhat disabled within a short period of 5 years after the onset of disease, and as many as 50% of patients within 10 years cannot work due to disability.
[0009]
Osteoarthritis exhibits symptoms similar to rheumatoid arthritis. In particular, rheumatoid arthritis develops as inflammation in synovial fluid, while osteoarthritis develops as a specific cartilage degeneration, but as the disease progresses, the processes become similar to each other. In osteoarthritis, reactive synovial fluid often appears as cartilage deteriorates and joint fit changes. Conversely, as rheumatoid arthritis erodes cartilage, secondary osteoarthritic changes in bone and cartilage appear. At the end stages of both osteoarthritis and rheumatoid arthritis, the affected joints have the same appearance.
[0010]
Some other forms of arthritis include ankylosing seronegative spondyloarthropathy (ankylosing spondylitis) and reactive arthritis. These medical conditions are often referred to as “B-27 related diseases” and are difficult to distinguish from rheumatoid arthritis. In some cases, ankylosing spondylitis, Reiter's syndrome or arthritic psoriasis coexist in the same patient with rheumatoid arthritis. In many cases, these patients are treated with the same disease ameliorating drugs as patients with advanced rheumatoid arthritis.
[0011]
The onset of arthritis generally occurs after 30 years of age in people who are vulnerable to such diseases. However, some types of arthritis can develop due to various causes such as delayed virus infection. Due to the large overlap, many physicians regard these types as “generalized rheumatism” and treat the disease equally. Some diseases that fall into this category include chronic fatigue syndrome, fibromyalgia (connective tissue inflammation) and gout. In fact, for some patients, evidence has accumulated that rheumatoid arthritis and fibromyalgia overlap.
[0012]
Autoimmune disease
As described above, rheumatoid arthritis is an autoimmune disease, and its etiology itself is almost the same as that of any other autoimmune disease. The normal body distinguishes between its own by-products and foreign invaders (ie, bacteria, viruses, filamentous fungi, protozoa, etc., to name a few). When immune cells (T or B lymphocytes) react with the developing “self protein”, they are considered defective and are usually killed or inactivated. However, sometimes it appears that “self-reactive” immune cells escape the killing. At some later time, the cells may be activated and cause an immune response. Activation is believed to occur after infection with common bacteria or viruses that contain a polypeptide having a stretch of amino acids that matches a stretch on a bad self-protein. Several bacteria such as Streptococcus, Mycoplasma, Borrelia, as well as certain viruses, ie retroviruses, have been implicated in the development of the disease. In addition to rheumatoid arthritis, autoimmunity often occurs in diseases such as juvenile diabetes, multiple sclerosis, Graves' disease, Menelli's disease, myasthenia gravis, lupus erythematosus, and psoriasis. (Medical Sciences Bulletin, September 1994).
[0013]
Autoimmunity affects specific organs. For example, some autoimmune diseases of the hepatobiliary and kidney include primary biliary cirrhosis, necrotizing glomerulonephritis, “idiopathic” crescent glomerulonephritis, viral liver and kidney disease, chronic hepatitis, self Immune and drug-induced hepatitis (Gershwin, Manns, and Mackay 1992). Diabetes was caused by immune destruction of the islets of Langerhans (Hagopian and Lermark 1992), and insulin autoantibodies were described (Palmer 1987).
[0014]
There is a large category of systemic vasculitis diseases that suggest autoimmune mechanisms as the etiology. Some of these diseases include leukocyte destructive vasculitis, nodular polyarteritis, Goodpasture syndrome, Kawasaki disease, Wegener's granulomatosis, Churg-Strauss syndrome, giant cell arteritis, Takayasu arteritis, immune Complex-mediated, wolf wounds, rheumatoid and cryoglobulinemia vasculitis, Henoch-Schönlein purpura (Kallenberg, 1996; Jennette, Jones, Talk, 1992).
[0015]
There is also a body of evidence that autoimmunity plays a role in a variety of heart diseases including pericardiotomy and post-myocardial infarction syndrome, myocarditis, idiopathic dilated cardiomyopathy. Autoimmunity appears to cause acute myocardial disease to progress to degenerative disease (Rose, Neumann, Burek, Herskowitz 1992).
[0016]
The involvement of skeletal muscle symptoms is (rheumatoid arthritis, rheumatic polymyalgia, myasthenia gravis, myasthenia myopathy, neurogenic atrophy, motoneuron disease, fibromyalgia, connective tissue inflammation, muscular dystrophy, endocrine It is common in many autoimmune diseases such as polymyositis or inflammatory myopathy (which may include sex, metabolic and cancerous myopathy). (Hollingsworth, Dawkins, Thomas 1992).
[0017]
Other autoimmune diseases are uveitis, Vogt-Koyanagi-Harada syndrome, (Detrick and Hooks 1992), and Sjogren's syndrome, scleroderma, ankylosing spondylitis, dermatomyositis, psoriasis, arthritic psoriasis, Reiter's syndrome (NIH 1994).
[0018]
Evidence for autoantibodies was also found in Alzheimer's disease (Singh et al., 1992), dementia complex (Mastroianni et al., 1991) and autistic children (Singh et al., 1993).
[0019]
Several neurological disorders, such as Sydenham chorea, chronic obsessive compulsive disorder (OCD), attention deficit hyperactivity disorder (ADHD), Tourette syndrome (TS), and some cases of schizophrenia have autoimmune components. And seem to be associated with anti-neuronal antibodies (Medical Sciences Bulletin, Sept. 1994).
[0020]
This summary is not exhaustive and the person skilled in the art is familiar with other autoimmune diseases such as, for example, Guyan-Barre syndrome (idiopathic polyneuropathy).
[0021]
Treatment
It is common medical practice to administer drugs that reduce the physical discomfort of the inflammatory response to treat inflammation-related disorders. Agents having these properties are classified as anti-inflammatory agents. Anti-inflammatory drugs are used to treat a wide range of disorders, and the same drugs are often used to treat various diseases. Treatment with anti-inflammatory drugs is most often directed to symptoms (ie inflammation) rather than to the disease.
[0022]
Anti-inflammatory, analgesic and antipyretic drugs are a heterogeneous group of compounds that are often chemically unrelated but nevertheless share certain therapeutic effects and side effects. Corticosteroids are a group of compounds most commonly used for the treatment of inflammation. Proteolytic enzymes are another group of compounds believed to have anti-inflammatory effects. Hormones that act directly or indirectly on the adrenal cortex to produce and secrete steroids are another group of anti-inflammatory compounds. Unfortunately, natural and synthetic corticosteroid formulations cause several severe side effects including elevated blood pressure, salt and water retention, kidney damage and increased potassium and calcium excretion. In addition, corticosteroids can mask the signs of infection and promote the spread of infectious microorganisms. These hormones are considered unsafe for use by pregnant women, and long-term treatment with corticosteroids has been associated with gastric hyperactivity and / or peptic ulcers. Treatment with corticosteroids can exacerbate diabetes, require higher doses of insulin, and cause mental disorders. Hormonal anti-inflammatory agents that indirectly increase the production of endogenous corticosteroids have the same potential for adverse side effects.
[0023]
Another common treatment for inflammation, particularly rheumatoid arthritis, other arthritis, and other autoimmune diseases is drug therapy. In general, patients are initially treated with “first use” agents, usually non-steroidal anti-inflammatory drugs (NSAIDs) that primarily relieve symptoms. Later, the patient is treated with a “second use” or disease ameliorating agent (DMARD) such as methotrexate, gold compounds, penicillamine, sulfasalazine, antimalarials. However, all of the drugs show serious side effects, especially when administered at high doses. For example, aspirin, a type of NSAID, can cause indigestion and stomach pain. Phenylbutazone can cause gastric ulcers, and phenacetin can cause kidney disease. Methotrexate can cause mouth ulceration and gastrointestinal (GI) side effects.
[0024]
If natural foods with anti-inflammatory effects can be obtained, they will provide an easily administrable, readily available and safe therapeutic composition for the treatment of arthritis, autoimmune diseases and inflammation in general. .
[0025]
Passive immunity
Various genera of birds, such as chickens (Gallus domesticus), turkeys and ducks, produce antigens that cause avian disease and antibodies against other antigens in blood and eggs. See, for example, LeBacq-Verheyden et al. (Immunology 27: 683 (1974)) and Leslie, G. et al. A. Et al. (J. Med. 130: 1337 (1969)) quantitatively analyzed chicken immunoglobulins. Polson, A et al. (Immunological Communications 9: 495-514 (1980)) immunized hens against a mixture of several proteins and natural proteins and detected IgY antibodies in egg yolk. Fertel, R.A. Et al (Biochemical and Biophysical Research Communications 102: 1028-1033 (1981)) immunized hens against prostaglandins and detected antibodies in the yolk. Jensenius et al. (Journal of Immunological Methods 46: 63-68 (1981)) provide a method for isolating egg yolk IgG for use in immunodiagnosis. Polson et al. (Immunological Communications 9: 475-493 (1980)) describe antibodies isolated from hen egg yolk immunized with various plant viruses.
[0026]
US Pat. No. 4,357,272 discloses antibody isolation from egg yolk from hyperimmunized hens. Hyperimmunity was induced by repeated injections of each antigen from plant viruses, human IgG, tetanus antitoxin, snake anti-benin and Serameba. U.S. Pat. No. 4,550,019 discloses the isolation of antibodies generated in hens from egg yolk by hyperimmunization with an antigen having a molecular or particle weight of at least 30000. Antigens used to hyperimmunize chickens were selected from plant viruses, human immunoglobulins, tetanus toxins and snake venoms.
[0027]
US Pat. No. 4,748,018 is a passive mammalian immunization method comprising parenterally administering a purified antibody obtained from avian eggs immunized against a corresponding antigen, the mammal Discloses a method of acquiring immunity to eggs.
[0028]
US Pat. No. 5,772,999, assigned to DCV-Biologics, provides chronic gastrointestinal disorders or non-steroidal anti-inflammatory drug-induced (NSAID) in a subject by administering to the subject a hyperimmune egg and / or milk or part thereof. Disclosed are methods for preventing, deterring or reducing (induced) gastrointestinal injury.
[0029]
US patent application 09 / 233,379 discloses the presence of anti-inflammatory factors found in eggs of hyperimmune birds.
[0030]
An immunized egg is, for example, an egg obtained from birds immunized with a specific antigen or antigen mixture. Hyperimmune eggs are eggs obtained from birds immunized to a specific immunity, for example, by periodic booster administration of antigen. Regardless of the type of antigen administered to the immunized bird, the hyperimmune egg can treat chronic gastrointestinal disorders, NSAID-induced gastrointestinal injuries (see US patent application Ser. No. 08 / 688,576) as described above. Have been found to have anti-inflammatory effects due to the presence of various beneficial factors, including, and the presence of anti-inflammatory compositions (see US patent application Ser. No. 09 / 233,379).
[0031]
Glucosamine
Glucosamine is a cartilage protective agent that has been studied for its potential beneficial effect on osteoarthritis. Chondroprotectors are agents that appear to help balance the synthesis and degradation of cartilage tissue as well as relieve symptoms. Glucosamine has been known to biologists for decades as an endogenous amino monosaccharide synthesized from glucose. Its importance in joint dysfunction is related to its physiological role in the synthesis of the cartilage components proteoglycan and glycosaminoglycan. Studies over the past 40 years suggest that glucosamine is effective and well tolerated in reducing the symptoms of joint dysfunction. Glucosamine significantly reduces pain and tenderness and improves motility. Researchers have proposed several mechanisms that explain the possible benefits of glucosamine in joint health. In vitro studies have suggested that glucosamine affects cartilage metabolism. One such effect appears to be the stimulation of proteoglycan synthesis, as is evident using SAMe. Another possibility is that glucosamine promotes chondrocyte gene expression. Moreover, glucosamine can act as an anti-inflammatory agent, but its action is 50 to 300 times lower than NSAID. However, NSAIDs exhibit anti-inflammatory effects through inhibition of the cyclooxygenase enzyme system and thereby blocking prostaglandins, whereas glucosamine does not. Its anti-inflammatory effect appears to stabilize the cell membrane as a result of stimulating proteoglycan synthesis. Recent in vitro studies have shown that glucosamine increases the adhesion of chondrocytes to fibronectin and effectively reverses the abnormal decrease in adhesion that occurs during joint degeneration.
[0032]
(Summary of Invention)
The present invention provides that when eggs and egg products, especially hyperimmune eggs or egg products, are combined with glucosamine and administered to a subject animal, such administration causes a reduction, sometimes even prevention, of inflammation and inflammation-related disorders in the subject animal. Further, this is based on the discovery of the present inventors that anti-inflammatory activity appears.
[0033]
In particular, the present invention is directed to compositions comprising glucosamine and egg products.
[0034]
The present invention is also directed to a method of reducing inflammation in a subject comprising administering to the subject an effective amount of glucosamine and egg product.
[0035]
Furthermore, the present invention is also directed to a method of reducing serum fibrinogen levels, comprising administering to a subject an effective amount of glucosamine and egg product.
[0036]
The invention further encompasses a method of reducing or preventing the onset of rheumatoid arthritis, comprising administering to a subject an effective amount of glucosamine and egg product.
[0037]
Finally, the present invention also encompasses a method of reducing or preventing the onset of osteoarthritis, comprising administering to a subject an effective amount of glucosamine and egg product.
[0038]
(Description of the invention)
The present invention relates to the discovery that the combination of egg products with glucosamine acts synergistically on inflammation and inflammation-related disorders.
[0039]
The following definitions apply throughout.
[0040]
Definition
The term “inflammation”, in the art-recognized sense, is a local protective reaction induced by tissue injury or destruction that also destroys, dilutes the cause of the injury and the injured tissue, or A sequestering response, a series of classic events of pain, fever, redness, swelling and loss of function, as well as increased permeability and blood flow, leaching of fluids containing plasma proteins, and leukocytes into inflammatory lesions It is used as a reaction that characterizes the histological involvement of a complex series of events, including dilatation of arterioles, capillaries and venules, with an uncontrolled and uncontrolled manner.
[0041]
The term “arthritis” means any of a variety of disorders characterized by inflammation and degeneration of connective tissue structures, particularly joints and related structures. Arthritis can be associated with pain, stiffness, or restricted mobility at these sites. Some forms of arthritis include rheumatoid arthritis, osteoarthritis, ankylosing seronegative spondyloarthropathy, reactive arthritis, chronic fatigue syndrome, fibromyalgia (connective tissue inflammation) and gout.
[0042]
The term “autoimmune disease” applies to standard medical definitions found in standard medical dictionaries such as Dorland and Taber. A description of various autoimmune diseases can be found in the background section of this document.
[0043]
The term “hyperimmunity” means exposure to one or more antigens such that the immune response is enhanced and maintained over an unexposed natural state.
[0044]
The terms “egg” or “egg product” mean any whole egg (table, hyperimmune or otherwise) or any product or part derived from that whole egg, respectively.
[0045]
The terms “table egg” or “table egg product” mean each whole egg or any product or part derived from that egg obtained from a spawning animal that is not maintained in a hyperimmune state.
[0046]
The term “hyperimmune egg” or “hyperimmune egg product” includes an anti-inflammatory composition obtained from laying animals maintained in a hyperimmune state and described in US patent application Ser. No. 09 / 233,379. , Mean whole egg or any product or part derived from that whole egg.
[0047]
The term “supernormal amount” means an amount that exceeds the amount found in eggs of laying animals that are not maintained in a hyperimmune state.
[0048]
The term “anti-inflammatory composition” refers to a composition disclosed in US patent application Ser. No. 09 / 233,379, wherein the composition interferes with or inhibits the inflammatory process.
[0049]
The term “partially pure egg anti-inflammatory composition” means an anti-inflammatory composition as described in US patent application Ser. No. 09 / 233,379.
[0050]
The term “combinatorially induced immunogen” refers to the process of generating molecular diversity within an immunogen by combinatorial synthesis.
[0051]
The term “biotechnological immunogen” refers to an immunogen obtained through gene cloning techniques and genetic engineering processes that allow for the insertion and translation of antigenic proteins.
[0052]
The term “genetic vaccine” refers to a nucleic acid vaccine that is generally produced by recombinant techniques and can induce an immune response.
[0053]
The term “treatment” delays or completely prevents the onset of symptoms (including pain) of the disorder and / or the pathogenesis of the disorder or, if any, improves or completely eliminates the symptom. It means to do. For example, the hyperimmune egg plus glucosamine product not only suppresses the symptoms of the disorder in humans and other mammals, but also acts as a prophylactic agent to interfere with the presence of the disorder in the recipient, thereby preventing arthritis and / or Or treat autoimmune diseases.
[0054]
The term “preventing” means reducing and / or eliminating the progression of the disease or eliminating the onset of the disease.
[0055]
The term “administering” means any method of delivering a substance to a subject by routes including oral, nasal, parenteral (intravenous, intramuscular, or subcutaneous), rectal, topical, or intraocular. .
[0056]
The term “animal” means the definition of the animal kingdom.
[0057]
The term “target animal” refers to an animal that functions as an egg or egg product producing animal.
[0058]
The term “subject animal” refers to an animal to which an egg or egg product produced by a target animal is administered.
[0059]
The term “immunogen” means a substance that is not immunologically acceptable but rather capable of eliciting a humoral antibody and / or a cellular immune response. This term means not only the product, ie the ability to react with antibodies, but also the ability to stimulate an immune response.
[0060]
invention
The present invention includes a combination of a hyperimmune egg product and glucosamine. In accordance with the principles of the present invention, a combination of an exogenous amount of a hyperimmune egg product and glucosamine is provided to a subject exhibiting an inflammatory response in order to reduce inflammation.
[0061]
In a preferred embodiment, the present invention provides hyperimmunity in combination with glucosamine that, when administered to a subject animal, reduces inflammation in the subject animal and is effective to treat and prevent arthritis and / or autoimmune diseases. Contains eggs or egg products. Hyperimmune eggs are preferably obtained from laying animals, more preferably from birds hyperimmunized with at least one immunogen. The hyperimmune egg plus glucosamine product is a product that is preferably administered orally to the subject animal. Furthermore, hyperimmunized eggs or egg products and / or glucosamine can be separated into more potent fractions that can be subsequently administered to the subject animal in various forms. Further, although preferably administered simultaneously, it is expected that glucosamine and egg product can be administered separately. When administered in combination with a subject, glucosamine and the table or hyperimmune egg product reduce inflammation by acting synergistically.
[0062]
Glucosamine
Glucosamine provides a major substrate for the synthesis of both collagen and proteoglycans. When glucosamine is administered to a subject, the production of additional amounts of collagen and proteoglycan can be utilized in the subject's natural healing process to repair connective tissue, so that glucose and glucosamine in the natural production by the subject of collagen and proteoglycan The rate limiting step is bypassed.
[0063]
Glucosamine is preferably in a salt form to facilitate delivery and uptake by the subject. Preferred salt forms are especially glucosamine hydrochloride and glucosamine sulfate. The use of non-salt glucosamine is believed by many to be unable to provide bioavailable glucosamine because this compound is not absorbed in the gastrointestinal tract. In the case of glucosamine sulfate, it has been recognized that this sulfate can be utilized in the subsequent catalysis of the conversion of glucosamine to glycosaminoglycans. Non-sulfate forms are desirable for the production of hyaluronic acid.
[0064]
With regard to the administration of glucosamine, oral administration allows it to be absorbed rapidly and almost completely by humans and other animals, with oral administration itself being most preferred. A significant portion of ingested glucosamine localizes to cartilage and joint tissue and stays in place for a long time. This indicates that orally administered glucosamine reaches connective tissue, where it is incorporated into newly synthesized connective tissue.
[0065]
Egg products
An egg product is any egg or egg fraction obtained from a laying animal. In a preferred embodiment, the egg product of the present invention is hyperimmune, etc., to produce, among other things, a superordinary amount of an anti-inflammatory composition (see US patent application Ser. No. 09 / 233,379) in eggs. Eggs obtained from manipulated birds.
[0066]
Superimmunity of laying animals
A hyperimmune egg or egg product can be produced by any egg-laying animal. It is preferred that the animal is a member of the bird cage, ie a bird. Poultry is preferred in the bird class, but other members of this class such as turkeys, ducks and geese are also suitable sources of hyperimmune egg products.
[0067]
When such an egg-laying animal is rendered specific immune status, for example by periodic booster administration of an immunogen, the animal, when consumed by a subject, contains a super-normal amount of an anti-inflammatory composition, in combination with glucosamine. When administered, it appears to produce eggs that exhibit beneficial properties that are effective in treating and preventing inflammation-related and autoimmune diseases in the subject.
[0068]
Even if birds are sensitized to various immunogens during normal immunization treatment for avian diseases and during normal exposure to environmental factors, the table egg contains a super-normal amount of egg anti-inflammatory composition. As shown by the fact that there is not, induction of immunosensitivity alone is not sufficient to cause such super-normal amounts to appear in eggs. It is only when the egg is in a particular hyperimmune state that it has the desired superordinary amount of anti-inflammatory composition.
[0069]
This special hyperimmune condition, which appears to be effective due to the inclusion of higher amounts of anti-inflammatory composition, is a sufficiently high dose of a specific immunogen or mixture of immunogens after the initial immunization It is preferably carried out by administering a periodic booster having The preferred dose of the booster should be at least 50% of the dose required to generate the primary immunity of the bird. Thus, there is a threshold booster dose below which the nature is not produced in the avian egg, even if the bird is normally immune. Knowing the requirements for developing and maintaining a hyperimmune state, varying the dose of immunogen depending on the genus and strain of spawning animal used to maintain the animal in a hyperimmune state is It is within the field of technology in the field.
[0070]
The hyperimmune state is preferably generated by any immunogen or combination of immunogens. The hyperimmune state is preferably carried out by multiple exposures to multiple immunogens, multiple exposures to a single immunogen, or a single exposure to an immunogen library. Nearly any immunogen can be used to induce a hyperimmune state, including but not limited to bacteria, viruses, protozoa, allergens, filamentous fungi or cells.
[0071]
Knowing the requirements for developing and maintaining a hyperimmune state, varying the dose of immunogen depending on the genus and strain of spawning animal used to maintain the animal in a hyperimmune state is It is within the field of technology in the field.
[0072]
In addition to immunization with natural immunogens, immunization may be performed using immunogens synthesized synthetically by combinatorial chemistry. The basic strategy is to assemble numerous combinations of chemical building blocks to generate diverse populations of molecules. Oligomers (Fodor, S. et al., Science 251: 767 (1991); Houghton, R. et al., Nature 354: 82 (1991)) and small organic molecules (Bunin, B. & Ellman, J., J. Am. Chem. Soc. 114: 10997 (1992)) several methods have recently been developed for solid-phase and solution-phase combinatorial synthesis of the library. Rapid multiple peptide and oligomer synthesis can serve as a source of combinatorial derived immunogens. Moreover, alternative strategies may allow organic building blocks to be added to the backbone molecule in a combinatorial fashion to improve immunogenicity.
[0073]
Instead of immunogenic vaccines, alternative ways of hyperimmunizing laying animals can be used, including the use of genetic vaccines. In particular, any DNA construct (generally consisting of a promoter region and an antigen coding sequence) appears to elicit an immune response. Genetic vaccines include antigen-encoding vectors, naked DNA fragments, plasmid DNA, DNA-RNA antigens, DNA-protein complexes, DNA-liposome complexes, DNA expression libraries, and viruses delivered to generate an immune response And bacterial DNA. Methods of DNA delivery include particle bombardment, direct injection, viral vectors, liposomes and jet injection, among others. When using these delivery methods, the required amount is much less and generally the persistence of immunogen production is high. When using such genetic processes, the preferred method of introducing DNA into birds is by intramuscular injection of DNA into the breast muscle.
[0074]
Methods of DNA delivery include, but are not limited to, particle bombardment, direct injection, liposomes and jet injection (Fynan, EF et al., Proc. Natl. Acad. Sci. USA 90: 11478-11482 (1993)). By replicating nucleic acids encoding known or unknown immunogens, promoter regions (especially CMV cauliflower mosaic virus) and SV40 bacterial sources in bacteria, plasmid DNA for use in DNA injection can be generated. Although several parenteral routes of administration of the DNA are effective in chickens, the preferred method is intramuscular injection into the thoracic muscle. The vaccine test is performed on laying birds, preferably chickens. The immunization is repeated at intervals of 1-2 weeks for up to 6 months.
[0075]
In general, the amount of DNA used for direct injection is preferably 50 to 300 μg of DNA in brine. 2.5M CaCl for particle bombardment ₂ 4-100 μg of DNA co-precipitated on gold beads by the addition of By this method of accelerating DNA-coated particles into living animals, immunization can be repeated within the skin.
[0076]
The following is a detailed description of a preferred procedure used to bring a laying animal to a highly immunized state in which the resulting hyperimmunized egg or egg product can be administered to a subject.
1. Select one or more immunogens.
2. An immune response is induced in laying animals by primary immunization.
3. In order to develop and maintain hyperimmunity, an appropriate dose of immunogen booster vaccine is administered.
4). Test the level of anti-inflammatory activity of hyperimmunized eggs.
5. Collect and process the eggs.
[0077]
A more detailed description of this procedure is as follows.
[0078]
Step 1: Any immunogen or mixture of immunogens may be used as a vaccine. The immunogen may be bacterial, viral, protozoan, filamentous, cellular, or any other substance that the laying animal's immune system is likely to react to. The key point at this stage is that the immunogen must be able to induce immunity and hyperimmunity in the laying animal. Although only a single immunogen can function as a vaccine for the method of the invention, one preferred vaccine is a mixture of bacterial and viral multivalent antigens selected from the following antigen groups. Bacterial antigens are enteric bacillus and bacteroides, pneumococci, pseudomonas, salmonella, streptococci, bacillus, staphylococci, neisseria, clostridium, mycobacteria, actinomycetes, chlamydia and mycoplasma. The viral antigen is preferably selected from the following antigen groups: adenovirus, picornavirus and herpes virus, although other viral antigen groups would be effective.
[0079]
In a preferred embodiment, a multivalent vaccine designated PL100 is used. The bacteria contained in the PL-100 vaccine are listed in Table 1 of Example 1. Both vaccines are available from DCV, Inc. Already assigned to U.S. Pat. No. 5,772,999 and U.S. patent application Ser. No. 09 / 233,379.
[0080]
Step 2: The vaccine can be a killed vaccine or a live attenuated vaccine, which can be administered by any method that induces an immune response. Preferably, immunization is performed by administering the immunogen by intramuscular injection. The preferred muscle for injection into birds is the chest muscle. A preferred dose is 0.05-5 milligrams of immunogenic vaccine. Other administration methods that can be used include intravenous injection, intraperitoneal injection, intradermal, rectal suppository, aerosol, oral, topical or intraocular administration. When using DNA technology for hyperimmunization, a much smaller amount, typically 300 micrograms, is required.
[0081]
Whether a vaccine induced an immune response in laying animals can be determined by several methods known to those in the immunology field. Examples of these methods include enzyme-linked immunosorbent assays (ELISA), antibody presence tests for stimulating antigens, and tests designed to assess the ability of host immune cells to react with antigens. The minimum dose of antigen required to induce an immune response depends on the vaccine treatment procedure used, including the type of adjuvant and the dosage form of the antigen used, and the type of egg-laying animal used as the host.
[0082]
Step 3: The hyperimmune state is preferably induced and maintained in the target animal by repeated administration of appropriate doses of booster at predetermined time intervals. A preferred time interval is 2-8 weeks over a period of 6-12 months. However, it is essential that booster administration does not produce immune tolerance. Such a process is well known in the art.
[0083]
Other hyperimmune maintenance procedures or combination procedures can be used, such as intramuscular injection for primary immunization and intravenous injection for booster injection. Still other procedures include co-administration of microencapsulated and liquid antigens, or intramuscular injection for primary immunization, and booster doses by oral or parenteral administration by microencapsulation. Several combinations of primary and hyperimmunization are known to those skilled in the art.
[0084]
Stage 4: It is appropriate to test the level of anti-inflammatory activity of the eggs. This can be done by any clinical and preliminary clinical evaluation that tests the effect of hyperimmunized eggs, egg products derived from them, on inflammation.
[0085]
Stage 5: This stage involves collecting and processing hyperimmunized eggs. Eggs can be collected by conventional methods. Egg processing can be performed in a variety of ways as described later in this document. Eggs can be further processed to purify the anti-inflammatory composition described in US patent application Ser. No. 09 / 233,379.
[0086]
Processing and administration
Hyperimmune egg products that can be subsequently administered to target animals in combination with glucosamine to treat inflammatory disorders by processing eggs collected from hyperimmunized animals, or fractions thereof containing partially purified anti-inflammatory compositions Is preferably produced.
[0087]
The egg itself, or a fraction thereof comprising the partially purified anti-inflammatory composition of the present invention, is combined with glucosamine by any means to treat or prevent inflammation including arthritis and / or autoimmune disease in the subject animal. Be administered. Administration is preferably performed by feeding the subject animal directly with an egg or any egg derivative in combination with glucosamine. It is important to realize that whole egg, egg yolk and egg white are non-toxic and safe natural food ingredients.
[0088]
In other embodiments, glucosamine and any fraction thereof comprising egg or partially purified anti-inflammatory composition is integrated as a dietary supplement. With particular regard to eggs, one preferred method of preparing an egg or any fraction thereof for introduction into a dietary supplement involves drying the egg into egg powder. Various methods for drying eggs are known, but spray drying is the preferred method. Methods for spray drying eggs are well known in the art.
[0089]
Such dried egg powder can be introduced into the beverage, for example, in the form of protein powder, strength enhancing beverages, protein supplements and any other nutritional sports related products. In addition, egg powder can be used for baking mixes, power bars, candy, cookies, and the like. Other egg processing examples include making omelets, soft-boiled or hard-boiled eggs, including fried eggs, or if desired, eating the eggs raw or processing them as liquid eggs.
[0090]
The inventors suggest that glucosamine and egg are preferably administered in combination, but do not need to be administered in the same form or as a product. For example, glucosamine is ingested in the form of a pill while the egg product is administered as a beverage.
[0091]
If it is desired to combine the partially purified anti-inflammatory composition with glucosamine, as described above, the partially purified anti-inflammatory composition can be purified by even further separation of the eggs, thereby parenterally, Other modes of administration would be possible, such as separate administration of egg products via subcutaneous, intravenous, intramuscular, abdominal, nasal, oral or topical. Moreover, it is believed that such additional separation can make encapsulated products and pharmaceutical compositions having the eggs or fractions thereof.
[0092]
Preparations for parenteral anti-inflammatory compositions include sterile aqueous or non-aqueous solutions, suspensions or emulsions. Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
[0093]
With particular reference to partially purified anti-inflammatory compositions, oral administration is preferably carried out by solid dosage forms including capsules, tablets, pills, powders and granules, among others. In solid dosage forms, the anti-inflammatory composition is mixed with at least one inert diluent such as sucrose, lactose, starch and the like. Such dosage forms can also contain additional substances other than inert diluents as usual. In the case of capsules, tablets and pills, the dosage form may comprise a buffer, a pH sensitive polymer, or any other delayed release capsule forming agent commonly used as an encapsulated composition in the food and pharmaceutical industry. Good. In addition, tablets and pills can be prepared with enteric coatings.
[0094]
Liquid dosage forms for oral administration of anti-inflammatory compositions include inert diluents commonly used in the pharmaceutical art and include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the inert diluent, the composition can also include wetting agents, emulsifying and suspending agents, and sweetening agents.
[0095]
Glucosamine and immunized egg product combination
Glucosamine and egg products act jointly as described above when administered simultaneously to a subject to reduce inflammation in the subject.
[0096]
With regard to administration of both glucosamine and immunized egg product, in one embodiment, an effective amount of glucosamine is administered to the subject regardless of the effective amount of the immunized egg product. In other embodiments, a composition comprising an effective amount of glucosamine and an immunized egg product is administered to the subject. Both modes of administration are likely to cause a synergistic effect.
[0097]
Administration to the subject should be carried out in an amount effective to treat or prevent inflammation. Preferably, a higher dose of glucosamine mixed with a lower dose of immunized egg product is administered to the subject. The opposite may be more preferable depending on the particular disorder, in which case such unequal doses are appropriate for treatment and prevention and should be administered in those amounts. Because of the cooperative action of these two compositions, it may be necessary to adjust the administration accordingly. Those skilled in the art are familiar with determining dosages that would best treat and prevent the target inflammation.
[0098]
The glucosamines and eggs of the present invention may be administered by any means that provides anti-inflammatory activity. For example, administration may be parenteral, subcutaneous, intravenous, intramuscular, abdominal, nasal or oral.
[0099]
It is preferred to carry out oral administration, particularly with solid dosage forms including capsules, tablets, pills, powders and granules. In solid dosage forms, glucosamine and egg products are mixed with at least one inert diluent such as sucrose, lactose, starch and the like. Such dosage forms can also contain additional substances other than inert diluents as usual. In the case of capsules, tablets and pills, the dosage form may comprise a buffer, a pH sensitive polymer, or any other delayed release capsule forming agent commonly used as an encapsulated composition in the food and pharmaceutical industry. Good. In addition, tablets and pills can be prepared with enteric coatings.
[0100]
Liquid dosage forms for oral administration of a combination of glucosamine and egg products include inert diluents commonly used in the pharmaceutical field and include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs . In addition to the inert diluent, the composition can also include wetting agents, emulsifying and suspending agents, and sweetening agents.
[0101]
Formulations of glucosamine and egg product combinations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions or emulsions. Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
[0102]
The dose of the active ingredient can vary. However, the amount of active ingredient should be such that a suitable dosage form is obtained. It will be appreciated that the dosage form selected will depend on the desired therapeutic effect, route of administration and duration of treatment.
[0103]
The dosage and frequency of administration will depend on the patient's age and general health, given the potential for side effects. Administration will also depend on concurrent treatment with other drugs and patient tolerance to the administered drug.
[0104]
Regarding administration of hyperimmunized eggs or egg products to a subject, the preferred dose range of hyperimmunized eggs or egg products to be administered to a subject has been determined to be from 100 milligrams to 10 grams per kilogram of subject weight. This is described in detail in the following examples.
[0105]
As for the partially purified anti-inflammatory composition itself, the preferred dose range of the partially purified composition purified and separated from whole eggs, yolk and egg white of hyperimmune eggs is 1 microgram to 400 milligrams per kilogram of subject weight. It was decided.
[0106]
Regarding administration of glucosamine to a subject, the preferred dosage range of glucosamine to be administered to a subject has been determined to be 10 to 5 grams per kilogram of subject weight and is detailed in the examples below. . Even more preferably, it is 100 milligrams to 2.5 grams relative to the target weight.
[0107]
In a preferred embodiment, a 50 lb (22.7 kg) subject with inflammation should be administered twice a day with a single tablet containing 250 mg glucosamine HCl, 750 mg hyperimmune egg product and 320 mg coated ester C. Continue administration as long as necessary to reduce or prevent inflammation. The duration and extent of treatment will depend on the improvement of the particular condition and subject condition.
[0108]
Examples of inflammatory conditions that can be treated by administration of the glucosamine and egg products of the present invention include rheumatoid arthritis, osteoarthritis, ankylosing seronegative spondyloarthropathy, reactive arthritis, chronic fatigue syndrome, fibromyalgia (connective tissue inflammation) And any form of arthritis, including but not limited to gout. Egg products of the present invention include rheumatoid arthritis, juvenile diabetes, multiple sclerosis, Graves' disease, Menelli's disease, myasthenia gravis, lupus erythematosus, psoriasis, systemic scleroderma, rheumatic fever, Shogren's syndrome; acute And subacute bursitis, acute non-specific tendonitis, systemic lupus erythematosus, systemic dermatomyositis, acute rheumatic carditis, pemphigus, bullous dermatitis, herpes, severe erythema, polymorphic exfoliative dermatitis, Cirrhosis, seasonal long-term rhinitis, bronchial asthma, ectopic dermatitis, serum disease, keratitis, iritis, diffuse uveitis (ureitis), vocal fold inflammation, optic neuritis, sympathetic ophthalmitis, symptomatic sarcoidosis, refller Syndrome, beryllium disease, hemolytic anemia, mastitis, mastoiditis, contact dermatitis, allergic conjunctivitis, arthritic psoriasis, ankylosing spondylitis, acute gout arthritis, herpes zoster chronic joint Rheumatic, osteoarthritis, is equally effective in any other degenerative joint diseases, for the treatment of autoimmune disease, such as any other related autoimmune diseases. In addition, a combination of glucosamine and egg products may be used to treat a person exposed to a potential inflammatory agent such as an allergen.
[0109]
Inflammation in general, whether it is a type of arthritis or an autoimmune disease, in the context of treatment and prevention of a particular disorder, combined with a hyperimmune egg product or any active fraction thereof including a partially purified anti-inflammatory composition Glucosamine is preferably administered to the subject in an amount that is immunologically effective to treat and prevent that particular disorder. The duration and extent of treatment will depend on the improvement of the particular condition and subject condition. The duration and extent of treatment, if present, will depend on the particular condition, the presence or absence of the condition, and if present, the progression of the condition in the subject. Glucosamine in combination with any active fraction thereof, including hyperimmune egg products or partially purified anti-inflammatory compositions, is provided in any amount that treats and / or prevents the pathology and signs of the pathology. For example, in some examples, daily doses ranging from less than one to several hyperimmune whole eggs (or hyperimmune egg products including equivalents of less than one to several hyperimmune whole eggs), Depending on the specific circumstances of the condition, the subject can be administered in combination with an effective amount of glucosamine. The more potent fractions can be separated and concentrated by the methods described herein as well as other known methods in the art.
[0110]
The dose of the active ingredient can vary. However, the amount of active ingredient should be such that a suitable dosage form is obtained. It will be appreciated that the dosage form selected will depend on the desired therapeutic effect, route of administration and duration of treatment.
[0111]
The advantageous properties of this invention can be appreciated by reference to the following examples that illustrate the invention.
[0112]
Example
Example 1
Preparation of PL-100 vaccine
A bacterial culture obtained from the American Type Culture Collection and containing the bacterial populations shown in Table 1 below was reconstituted in 15 mL of medium and incubated at 37 ° C. overnight. After sufficient growth was obtained, about half of the bacterial suspension was used to inoculate 1 liter broth with inoculated bacteria incubated at 37 ° C.
[0113]
After observing sufficient growth in the culture, the bacterial cells were harvested by centrifuging the suspension for 20 minutes to remove the medium. The resulting bacterial pellet was resuspended in a sterile saline solution and the bacterial sample was centrifuged three times to wash off the media from the cells. After three sterile saline washes, the bacterial pellet was resuspended in a small amount of double distilled water.
[0114]
The medium-free bacterial suspension was sterilized by placing the suspension in a glass flask overnight in an 80 ° C. water bath. The viability of the broth culture was tested with a small amount of dead bacteria, incubated at 37 ° C. for 5 days, and examined daily for growth to ensure that the bacteria were already dead.
[0115]
The dead bacteria were lyophilized until dry. The dried bacteria are then mixed with a sterilized aqueous salt solution and 2.2 × 10 ⁸ The concentration was bacterial cells / mL saline (absorbance reading 1.0 at 660 nm). The bacteria contained in the PL-100 vaccine are listed in Table 1 below.
[0116]
[Table 1]

[0117]
Immunization procedures to obtain hyperimmune egg products
A pathogen-killed preparation was prepared as described above. For the initial vaccine treatment, the bacteria were mixed with complete Freund's adjuvant and 5.6 mg of bacterial material was injected into the chicken breast muscle. For subsequent vaccine treatment, the bacterial preparation was mixed with incomplete Freund's adjuvant and injected into chickens at 2-week intervals for 6 months.
[0118]
Example 2
the purpose
To assess the joint anti-arthritic effects of glucosamine hydrochloride and DCV hyperimmune eggs in established rat adjuvant arthritis, a chronic animal model of inflammation.
[0119]
result
Table 2 shows all data regarding paw edema on days 14, 22, and 30 of the study. The results show that the PL-100 + glucosamine HCl group showed the highest percent inhibition of inflammation on the 14th and 30th days of the disease. PL-100 eggs and glucosamine HCl showed inhibitory effects alone on the 14th and 22nd day of the study, but were ineffective on the 30th when the disease progressed rapidly to severe polyarthritis.
[0120]
Table 3 shows all serum fibrinogen levels on days 16, 23 and 30 of the study. From the results, it can be seen that the amount of fibrinogen first increases on the 16th and then decreases on the 23rd and 30th. However, the 16 day increase is minimal in the PL-100 + glucosamine HCl group compared to PL-100 and glucosamine HCl alone, and the inhibition of serum fibrinogen levels is also compared to PL-100 and glucosamine HCl alone. , PL-100 + Glucosamine HCl group is the highest. On the 30th, the percent inhibition rate of fibrinogen amount in the PL-100 + glucosamine HCl group is 32%, compared with 46% in the indomethacin treatment group.
[0121]
Table 4 shows all histomorphological observations on days 22 and 30 of the study. The results show that peri-articular / soft tissue inflammation (indicating mononuclear and / or polymorphonuclear inflammatory cell infiltration into soft tissue, including peri-articular synovium) was observed on both 22 and 30 days. It can be seen that the PL-100 + glucosamine HCl group has the least effect compared to the group of No ..
[0122]
Hyperostosis (indicating an increase or excess of new bone growth, especially in the subperiosteal area) is minimal at 22 days in the PL-100 + glucosamine HCl group and small at 30 days compared to the other groups.
[0123]
Day 22 and 30 cartilage erosion is minimal in the PL-100 + glucosamine HCl group compared to the other groups.
[0124]
Pannus cartilage (indicating accumulation of inflammatory cells, exudate, fibrin or fibrous tissue at the joint surface) has the least effect in the PL-100 + glucosamine HCl group on days 22 and 30 compared to the other groups.
[0125]
(Fibrous and inflamed tissues are formed between the joint surfaces, and the normal cartilage is completely lost, and this area is replaced with fibrous tissue, resulting in enlargement and loss of mobility that can result in adhesion and joint space. Ankylosia (representing a joint) is minimal in the PL-100 + glucosamine HCl group compared to any other group.
[0126]
Finally, osteolysis (indicating bone necrosis and lysis) is absent at 22 days in the PL-100 + glucosamine HCl treatment group compared to other groups where some lysis occurs.
[0127]
[Table 2]

[0128]
[Table 3]

[0129]
[Table 4]

[0130]
Summary
In a rat adjuvant arthritis model, which is a chronic animal model of inflammation, PL-100 egg and glucosamine HCl showed additional effects in the following respects.
a) Decreased swelling of the animal's rear left paw, thereby showing anti-inflammatory effects on days 14, 22 and 30.
b) Twenty-two days after injection, the amount of serum fibrinogen (which is a serum marker with inflammation) was reduced and continued until day 30 of the study.
c) On the 22nd day when the degree of rheumatoid arthritis reached its peak, the effects on the periarticular / soft tissues due to infiltration of mononuclear and polymorphonuclear cells were minimized.
d) Complete protection from hyperostosis on the 22nd day of the study.
e) Fully protected against cartilage erosion around the arthritic joint on day 22 and minimized injury on day 30.
f) The cartilage in the pannus area was fully protected on day 22 and the injury was minimized on day 30.
g) Fully protected the joints affected by ankylosing on the 22nd and fully protected the bones in the arthritic joint area from osteolysis on the 22nd.
h) FIG. 1 shows all data for foot edema on days 14 and 22 of the study. The results reveal that the PL-100 + glucosamine HCl group showed the highest percent inflammation inhibition rate on day 14 of disease.
i) FIG. 2 shows all serum fibrinogen levels on days 16, 23 and 30 of the study. From the results, it can be seen that the amount of fibrinogen first increases on the 16th and then decreases on the 23rd and 30th. However, the increase in 16 days is minimal in the PL-100 + glucosamine HCl group compared to PL-100 and glucosamine HCl alone, and the inhibition of serum fibrinogen levels is also compared to the PL-100 and glucosamine HCl alone groups. It is the highest in the PL-100 + glucosamine HCl group. On the 30th, the percent inhibition rate of fibrinogen amount in the PL-100 + glucosamine HCl group is 32%, compared with 46% in the indomethacin treatment group. Thus, the PL-100 + glucosamine HCl group not only suppresses the severity of inflammation at onset, but also blocks inflammation towards the end of the study (as indicated by the serum inflammation marker level).
[Brief description of the drawings]
FIG. 1 is a bar graph showing the effects of hyperimmunized eggs and glucosamine in a rat adjuvant arthritis model.
FIG. 2 is a bar graph showing plasma fibrinogen levels in a rat adjuvant arthritis model.

Claims (8)

A composition for treating arthritis, comprising glucosamine and a hyperimmune egg product.   The composition of claim 1, wherein the glucosamine is selected from the group comprising glucosamine HCl and glucosamine sulfate.   3. Composition according to claim 1 or 2, characterized in that the egg product is selected from the group comprising whole eggs, egg yolk, egg white and any fractions thereof.   4. Composition according to any one of claims 1 to 3, characterized in that the egg product is obtained from an egg-laying animal that is immunogenic or hyperimmunized with a genetic vaccine.   5. The immunogenic vaccine comprises at least one antigen selected from the group consisting of bacterial, viral, protozoan, fungal and cellular immunogens, and mixtures thereof. The composition as described.   The genetic vaccine is selected from the group consisting of naked DNA, plasmid DNA, viral DNA, bacterial DNA, DNA expression library, DNA-RNA immunogen, DNA-protein complex and DNA liposome complex fragments, and mixtures thereof 5. The composition of claim 4, comprising at least one immunogen-encoding DNA structure that is prepared.   The amount of egg product comprises a fraction of whole eggs in an amount between or equivalent to 100 milligrams of whole eggs per kilogram of subject weight to which the composition is administered. 7. The composition according to any one of 6 above.   The composition according to any one of claims 1 to 7, wherein the amount of glucosamine comprises from 10 milligrams to 5 grams.

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