TWI221772B - Chicken egg anti-inflammatory composition in partially purified form - Google Patents

Abstract

The invention is directed to a composition and method for the treatment and prevention of inflammation in general. The composition is an egg product obtained from an avian which has been hyperimmunized with no immungenic mixture. The invention is further directed to an anti-inflammatory composition which has been partially purified from a hyperimmunized egg. The partially purified anti-inflammatory composition, when administered to a subject animal is effective in treating and preventing inflammation.

Description

1221772 A7 V. Description of the invention (1) The present invention relates generally to compositions and methods for the treatment and prevention of inflammation, and the present invention is particularly related to an immediate-source anti-inflammatory composition, a method of preparing it in a partially purified form, and its use in treating inflammation ( Especially arthritis), and autoimmune diseases. Inflammation, as defined in the DodaruTs MecUcal Dictionary, is a local protective response that is stigmatized by tissue injury or destruction, which is used to destroy, weaken or block injury agents and injured tissue. "It is characterized by microvessels Perforation, blood components penetrate into the interstitial space, and white blood cells move into the inflamed tissue. When viewed from the giant, it is usually accompanied by redness, edema, hyperalgesia (softness), and pain. In this complex reaction, it is local Chemical mediators are released, such as histamine, 5-hydroxytryptamine, various chemotactic compositions, bradykinin, leukotrienes, and prostaglandins. Phagocytic cells will move to this area and the cell AS The membrane may rupture and release cytolytic enzymes. Inflammation derived from rheumatoid arthritis is generally associated with antibody complement 4, which promotes the local release of chemotactic and chemically active composition I, which attracts white blood cells. White blood cells Will chew the complex formed by antigen-antibody and complement, and also release many enzymes contained in lysosomes. Will promote damage to cartilage and other tissues, and this will increase the degree of inflammation. In addition, it may also involve the immune response through the cell-mediated. In this process, prostaglandins are also produced, which is the main intracellular regulation of cell functions. Please read back 1¾ 5 i matters first

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Disaster response is any reaction that has the inflammatory characteristics defined above. It is well known to those skilled in the medical arts that the inflammatory response causes many physical discomforts (ie, pain and loss of function) associated with different diseases2 injuries. ETQ / r / r Guan Jiyan Guan Yinyan itself has various forms. Some of the more common forms include rheumatoid arthritis, osteoarthritis and generalized rheumatism. Rheumatoid arthritis is an autoimmune disease characterized by joint pain, swelling and stiffness, and rheumatoid arthritis is a disease that causes about 3% of Americans', especially women, to suffer. Rheumatoid arthritis is a severely disabled disease that usually affects adults between the ages of 30 and 40, with the highest incidence of clinical signs between the ages of 40 and 60. Although drug treatment is very effective ', as many as 7% of rheumatoid arthritis patients are rapidly disabled to a considerable extent within 5 years after the onset of the disease, and as many as 50,000 / 0 are disabled to be unable to work (Medical Sciences Bulletin , December 1994). Printed by the Consumers' Cooperation Program of the Central Bureau of Standards of the Ministry of Economic Affairs Osteoarthritis produces symptoms similar to rheumatoid arthritis. In particular, although osteoarthritis starts from the degeneration of articular cartilage and rheumatoid arthritis starts from the inflammation of the synovium of the joint, as the disease progresses, the procedures gradually approach each other. In osteoarthritis, reactive cartilage synovitis often develops with the deterioration of cartilage and changes in joint engagement. In contrast, as rheumatoid arthritis invades cartilage, secondary osteoarthritis changes develop in bone and cartilage. The joints involved in the final stages of both osteoarthritis and rheumatoid arthritis will appear the same. There are other forms of arthritis, including sero-negative spondylosis of the joints (joint spondylitis) and reactive arthritis. These conditions are often referred to as -5- This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) 1 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 ^^ -------- B7 _ V. Description of the invention (3) B-2 7 related diseases) are difficult to distinguish from rheumatoid arthritis. In some cases, joint adhesion spondylitis, Reiter's microclimate, or psoriatic arthritis can coexist with rheumatoid arthritis in the same patient. In many cases, their patients were treated with the same disease-modifying drugs as those with progressive rheumatoid arthritis. The onset of arthritis usually occurs after the age of 30 in those who are susceptible to the disease. However, some forms of arthritis may be initiated by different causes, such as chronic viral infections. Because there is a substantial overlap with each other, many physicians treat their forms as ', generalized rheumatism' and treat them in the same way. Some diseases falling into this category include chronic fatigue microclimates, Fibromyalgia (fibromyalitis) and gout. In fact, for some patients, evidence has accumulated to show an overlap between rheumatoid arthritis and fibromyalgia (Harris,

Edward D. and W.B. Saunders, Rheumatoid Arthritis, 1997). Autoimmune Diseases As mentioned above, rheumatoid arthritis is an autoimmune disease, so its etiology is quite the same as that of any other autoimmune disease. Normally, the body normally recognizes the differences between its own byproducts and foreign invaders (ie, bacteria, viruses, fungi, and protozoa, for example). When an immune cell (T or B lymphocyte) responds to, and autologous protein " in its development, the cell is considered defective and is usually destroyed or deactivated. However, sometimes “autoreactive” immune cells escape from being destroyed. After a certain period of time, the cells can be activated and initiate an immune response. In general, the activation occurs after common bacterial or viral infection, which has a segment of amine group-6 on the defective autoprotein-This paper size applies the Chinese National Standard (CNS) Α4 specification ( 210X297 mm) --- (Please read the notes on the back before filling this page)

1221772 A7 _______ B7_ V. Description of the invention (4) The fe chain coincides with a segment of amino acid chain. There are several types of bacteria, such as Slreptococcns, Mycoplasma, and Borrelia, and certain viruses (retroviruses) are involved in the onset of disease. In addition to rheumatoid arthritis, autoimmunity often causes diseases such as juvenile diabetes, multiple sclerosis, Graves' disease, Meneri's disease, myasthenia gravis, post-erythema therapy, and psoriasis (Medical Sciences Bulletin, September , 1994). Autoimmune affects specific organs. For example, some autoimmune diseases of the hepatobiliary ducts and kidneys are: primary biliary sclerosis, necrotizing glomerulonephritis, 'spontaneous crescent glomerulonephritis, virus, liver and kidney disease, chronic Hepatitis, autoimmune hepatitis, and drug-induced hepatitis (Gershwin, Manns, and Mackey 1992). It has also been reported that the immune disruption of Lang's Island causes diabetes (Hgopian and Lernmark 1 " 2) and insulin autoantibodies (Palmer 1987). Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs. A large class of systemic vascular disease is presumed to be caused by autoimmune mechanisms. The components of this type of disease are: leukocyte vasculitis, polyarteritis inflammation, Goodpasture's syndrome, Kawasaki disease, Wegner's granulomatosis' Churg-Struass syndrome, giant cell arteritis, Takayasu arteritis 'Immune complex mediators' lupus, rheumatism, and cryoglobulinemia vasculitis, Henoch Schonlein purpura (Kallenberg, 1996; Jennette, Jones, Falk, 1992) ° There is also much evidence that autoimmunity may Forms of heart disease have a role, including: posterior pericardial incision and posterior myocardial infarction syndrome, myocarditis, and spontaneous dilated cardiomyopathy. Autoimmune may also contribute to acute heart-size paper. Applicable to China National Standard (CNS) A4 (210x 297). Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 1221772 A7 B7 V. Description of the invention (5) Derivation of muscle diseases (Rose, Neumann, Burek, Herskowitz 1992). Symptoms and symptoms are common to many autoimmune individuals, such as polymyositis or inflammatory myopathy (which may include rheumatoid arthritis, polymyalgia rheumatism, myasthenia gravis, muscles Weakness-type myopathy, neurotrophic atrophy, motor neuron disorders, fibromyalgia, fibrous tissue inflammation, muscular dystrophy 'endocrine' metabolic and cancerous myopathy) (Hollingsworth, Dawkins, Thomas 1992) Other diseases of autoimmune origin can be uveitis, Vogt-Koyanagi-Harada syndrome (Detrick ans Hooks 1992), and Sj6gren's syndrome, scleroderma, arthritis spondylitis, dermatomyositis, psoriasis, skin Psoriasis arthritis, Reiter's syndrome (NIH 1 994). Signs of autoantibodies also appear in Alzheimer's disease (Singh et al., 1992), relapses in dementia (Mastroianni et al., 1991, and autism) (Singh et al, 1993). There are several Neurological diseases such as Schodenmo's chorea, chronic obsessive-compulsive disorder and obsessive-compulsive behavior disorder (0CD), attention deficit hyperactivity disorder (ADHD), Tourette's syndrome (D S) and some schizophrenia may have Immune components and may be associated with antibodies against a nerve (Medical Sciences Bulletin, September, 1994). The above summary does not cover all, and those skilled in the art are familiar with other autoimmune diseases, for example, Guillain- Barr6 syndrome (spontaneous fissure polyneuritis). Treatment In order to treat inflammation-related diseases, the common medical operation is to take 矸 minus -8-This paper size applies the Chinese National Standard (CNS) A4 size (210X297 mm) (please first Read the precautions on the reverse side and fill out this page). Order 1221772 A7 V. Description of the invention (6 Medicines with physical discomfort. Distribution of medicines of this nature ... Anti-inflammatory drugs can be used for treatment Range of diseases, and ~, use the same drug to treat different diseases. Use anti-inflammatory drugs two? Target the disease, but often the symptoms (ie, inflammation). Treatment is not resistant: inflammatory drugs' analgesics, and withdrawal Fever medicine is not: Hua Er Chang often is irrelevant, but there are certain therapeutic effects. 222 shellfish steroids represent the most widely used class of compounds to treat inflammation. Egg knife enzymes represent another-class that is considered to have anti-inflammatory properties. Compounds that work. Hormones that directly and indirectly cause the production and secretion of steroids in the renal cortex represent another 2 inflammatory compounds. Unfortunately, 'natural, and synthetic corticosteroid preparations can cause many serious side effects, including increased blood pressure, salt and water Retention, kidney injury, and accelerated excretion of potassium and dance. Furthermore, corticosteroids may mask signs of infection and promote the spread of infectious microorganisms. These stimuli # cannot be safely used in pregnant women and long-term corticosteroids :: gastric overactivity And / or gastric ulcers. Treatment with corticosteroids may also exacerbate diabetes, require higher doses of insulin, and may Spermatogenic disease. Those who indirectly increase endogenous corticosteroids in hormonal anti-inflammatory drugs have the same potential for adverse side effects. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs for inflammation, and especially rheumatoid arthritis Another commonly used treatment for other arthritis and its autoimmune diseases is drug therapy. Generally, 1 *, patients are used at the beginning, and the first-line "medicine" is often a non-medical drug that mainly relieves symptoms. Steroid anti-inflammatory drugs (NSAIDS) are treated. Patients are subsequently treated with "second-line" or disease mediators (DMARDS), such as methotrexate, gold compounds, penicillinamine, sulfasalazlne, and this paper Applicable to China National Standard (CNS) A4 specification (2 丨 0χ297 public shame 72 7 11 2 12 A7 B7 V. Description of the invention (7) Anti-paeonia. However, all of these drugs have severe side effects, especially when taken in high doses. For example, aspirin, an NS AID, can cause indigestion and stomach pain; phenylbutazone can cause gastric ulcers, but not phenacetin, which can cause kidney disease. Methotrexate may cause oral ulcers and gastrointestinal (GI) side effects. If natural foods with anti-inflammatory effects are available, they can provide easily administered, readily available, and safe therapeutic compositions for the treatment of arthritis, autoimmune diseases, and general inflammation. The genus of birds, such as chickens (Gallus domesti cus, fire disaster, and ducks), produced by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs will produce immunogens in blood and eggs that fight poultry diseases and other immunogens Antibodies such as LeBacq-Verheyden et al. (Immunology 27: 683 (1994)) and leslie, GA. Et al. (J. Med. 130: 1337 (1969)), have quantitatively analyzed difficult immunoglobulins Protein. Dolson, A. et al. (Immunological Communications 9: 495-514 (1980)) immunized the mother against several proteins and natural protein mixtures, and detected IgY antibodies in the yolk of eggs. Fertel. R. et al. (Bio-chemical and Biophysical Research Communications 102: 1028-1033 (1981)) immunized hens against prostaglandins to detect antibodies in yolk. Jensenins et al. (Journal of Immunological Methods 46: 63-68 (198 1)) proposes a method for isolating yolk IgG for immunodiagnosis. Poison et al. (Immunological Communications 9: 475-493 (1980)) addresses the difficulty of immunizing mothers treated with various plant viruses. Yolk isolated U.S. Patent No. 4,3 5 7,2 7 2 discloses the isolation of antibodies from the yolk of ovum produced by hyperimmune mothers. This hyperimmune system is derived by repeated injections. National CNS of China) A4 size (210 \ 297 mm1) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 1222172 A7 B7 V. Description of the invention (8) From plant viruses, human IgG, tetanus antitoxin, anti-venom serum, and Serameba The immunogen was defamated. U.S. Patent No. 4,55,0019 discloses the isolation of antibodies from the hard-to-female eggs 5 that are hyperimmunized with an immunogen having a molecular weight of at least 30,000 or a particle weight. . The immunogen used for difficult hyperimmunization is selected from the group consisting of plant virus, human immunoglobulin, tetanus antitoxin, and snake venom. US Patent No. 4,748,0 18 discloses a passive immune treatment for mammals, including A parenterally administered purified antibody obtained from egg I that has been immunized against a relative antigen, and the mammal acquires immunity to the egg. US Patent No. 5,772,999 (assigned to DCV-Bi. 1〇gic s) A method for preventing, combating or reducing chronic gastrointestinal disease or non-steroidal anti-inflammatory drug-induced (NSAID-induced) gastrointestinal injury in a patient, which comprises administering the patient with hyperimmune eggs and / or milk or parts thereof Serving. However, there are no general references to reveal or speculate that eggs when administered to animals have the power to prevent or reduce inflammation 'to treat or prevent arthritis or to treat or prevent autoimmune diseases. Moreover, these references do not reveal or speculate a method that could reasonably be expected to result in hyperimmunization of poultry that would produce poultry that would lay eggs with this ability. Finally, these references do not disclose or speculate that poultry's seals would contain anti-inflammatory compositions. The present invention is based on the findings of the inventors of the present case, that is, eggs and egg products contain anti-inflammatory activity, especially egg products obtained from hyperimmune birds, which are given to individual animals, especially mammals, when taken. , Predictable

/ 72 Five 'Invention Description (9 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs to prevent or reduce the inflammation of their animals. In particular, the present invention is related to inflammation compositions. The anti-inflammatory composition n I ~ The present invention also relates to a kind of hyper-immune immune system, which can produce an anti-inflammatory composition with ultra-positive levels in poultry eggs. Anhua 4w this month Human L is the level of anti-inflammatory composition in egg white and protein via this hyperimmunization. The present invention also encompasses a method of treating a patient who is not mammalian, including administering to the patient a portion of pure anti-inflammatory Inflammatory composition or a composition containing the partially pure anti-inflammatory composition. This part covers taking the whole egg itself and / or a part thereof. The present invention is further based on the findings of the inventor of the present case, that is, in eggs and eggs It is active in the product, especially when it is obtained from a hyperimmunized poultry product, which can be used to treat or prevent arthritis and / or autoimmune diseases of an individual animal, especially a mammal. H ^ Then, the present invention relates to a method in which an egg product is administered to an individual animal in combination with a drug far from a combination of a non-steroidal anti-inflammatory drug and a disease-modulating anti-arthritis drug. Brief Description of the Drawings Figure 1 is Flow chart of purifying anti-inflammatory composition with molecular weight below 3,000 from powdered egg yolk. Figure 2 is a flow chart of purifying anti-inflammatory composition with molecular weight below 3,000 from powdered egg white. Figure 3 In order to separate the partially purified anti-inflammatory composition with a Sephor arse column (please read the precautions on the back first and then the first page)

IH ·· —ϋ I -Order-12 This paper size is applicable to China National Standard (CNS) A4 specification (210X 297mm) 1221772 5. Description of the invention (10 Absorbance chart printed by the Consumer Cooperatives of the Central Standards Bureau, Ministry of Economic Affairs. Θ is the Η PLC chromatogram of the partially purified anti-inflammation composition isolated from the fiery hyperimmune yolk. The two L is a graph showing the effect of hyperimmune eggs on the occurrence of guan ji inflammation using a mouse type 11 collagen model. Mo = 6 is a graph showing hyperimmune eggs, some of which are partially and partially purified anti-inflammation, and e-substances on the severity of arthritis in a rat collagen type 丨 丨 model. "A detailed description of the invention The invention relates generally to compositions and methods for the treatment and prevention of generalized inflammation, and in particular to inflammation and autoimmune diseases. The composition is preferably a natural food product including hyperimmune eggs or egg products. The food product is When administered according to the method of the present invention, it not only provides the effect of reducing pain and other microclimates caused by arthritis and autoimmune diseases, but also delays and even prevents the onset of those diseases. Beginning. Injections into birds to produce hyperimmune egg products are free of specific antigens known to cause inflammation or autoimmune diseases. So, surprisingly, poultry immunized from mixed antigen vaccines The resulting hyperimmune eggs or egg products can effectively reduce the symptoms of inflammation and autoimmune diseases and prevent them from occurring when administered to patients. Definitions The following definitions apply throughout this article: "Inflammation" Its technically accepted meaning refers to a local protective response issued by tissue injury or destruction for destruction, dilution or -13- This paper size applies to China National Standard (CNS) A4 (210X 297) Mm) Please read the note on the back of I. This page

I A7 B7 V. Description of the invention (11) In the pure formula for clogging the wounding agent and the injured phase, it is characterized by: 2: the second is its improper, typical sequence of uncontrolled loss, and 1 ::! , Redness, swelling, and function include arterioles, microvessels, and II-a complex series of events, including within the source site. 1. Spoon exudates, and white blood cells move to inflammation. Phase ::: two: H means to show connective tissue structure, especially joints and: to fire and k sex <any kind of disease. It may be accompanied by pain, stiffness, and dysfunction. Certain arthritis materials include rheumatism = osteoarthritis' bone and joint a, joint adhesion seronegative spondylopathy, reactive arthritis' chronic fatigue microclimate 'fibromyalgia (fibritis) and gout. &quot; Autoimmune diseases are used in standard medical dictionaries such as Doriand, s and abers. A description of various autoimmune diseases can be found in the "Background," paragraph of this document. Hyperimmunization-consent means that exposure to one or more antigens increases the immune response and maintains the natural unexposed state. The term "egg or egg product" refers to any whole egg (tabk) egg, any product or part thereof derived from or printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs, printed by a hyper-immunized or otherwise. The terms "edible egg" or "edible egg product" each refer to a whole egg or any product or part thereof derived from an oviparous animal that has never remained in a hyperimmune state. The terms "hyperimmune egg" or "hyperimmune egg product" each refer to a whole egg or any product or part thereof derived from an oviposition animal maintained in a hyperimmune state. -14-This paper size applies towels @National Standards (CNS) M specifications (21Qx297 Gongchu 1221772 Α7 Β7 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economy Levels measured in eggs produced by egg-laying animals in a hyperimmune state. "Anti-inflammatory egg or egg part body," the term means an egg or egg part containing an anti-inflammatory composition as shown herein "An anti-inflammatory composition" means a composition of the present case that can counteract or suppress the inflammatory process. "Shao Fen's pure egg anti-inflammatory composition" means a compound having at least Examples and their exemplified substances and figures. An anti-inflammatory composition of the purity described in the formula (Supplementary Note). "Combined-derived immunogen" refers to the process of generating molecularly differentiated immunogens by combinatorial synthesis. The term "bioengineered immunogens" refers to genetic selection Method and genetically-immunized protein obtained by embedding and translating proteins with antigenic properties. Gene vaccines π — The term refers to those that are usually produced by recombinant technology and can be induced A nucleic acid vaccine for disease response. • The term 'treatment' means that the onset of disease symptoms (including pain) and / or the cause of the disease is delayed or completely suppressed, or, if present, the symptoms are reduced or eliminated altogether. For example, hyperimmune egg products can treat arthritis and / or autoimmune diseases. They can not only suppress the disease microclimate of humans and other animals, but also act as a preventive agent to offset the presence of disease in the recipient. Prevention-consent refers to disease The progress of the disease is slowed and / or eliminated, or the onset of the disease is eliminated. The term "administration" means any method of providing a substance to a patient, including the application of Chinese National Standard (CNS) A4 specifications (-) 210X 297 mm) (Please read the notes on the back before filling out this page) r Pack_: Order Α ^ ι / 72 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (13) Non-intestinal (Intravenous, intramuscular, or subcutaneous), rectal, topical, or intraocular. The term "animal" means the definition of the animal kingdom. "Animal" means an animal that produces an animal as an egg or egg product. ' The term "individual animal" refers to an animal that uses eggs or egg products produced by the target animal. "The term" immunogen "refers to the ability to elicit humoral antibodies and / or cellular-mediated immune responses and Substances that are susceptible. 敫 &amp; shows the ability to react and react with its products (ie, antibodies). The products and methods of the present invention are particularly relevant for the treatment and prevention of inflammatory and autoimmunity in natural food hyperimmune eggs. Uses on sexual diseases. Since they are natural, this kind of mouth can be used to treat and prevent those diseases. Of course, in addition to allergic reactions to people with egg intolerance, they are not afraid of side effects. In the present invention, the present invention includes hyperimmune eggs or egg products, which can effectively treat and prevent arthritis and / or autoimmune diseases in individual animals. The hyperimmune egg line is obtained from maggots, and more particularly birds, which are hyperimmune with soil ν immunogens. The hyperimmune egg product is preferably administered to an individual animal orally. This hyperimmune egg or egg product can be more isolated into more laxative #trauma, which can be subsequently administered to individual animals in various forms. In another example, the present invention includes an anti-inflammatory composition obtained from poultry eggs, partially purified, and a partially purified composition administered to a patient to treat hair. CNS) mm (please read the notes on the back before filling this page)

221772 A7 B7 V. Description of the invention (14 Please read the notice on the back and then fill in the apical syringitis. The present invention also includes the use of one or more immunogens, and in particular, bacterial antigens, or their synthesis, etc. The same anti-inflammatory composition is obtained by partially purifying the poultry eggs that have been hyperimmune with the effective substance. The anti-inflammatory composition is present in the hyperimmune eggs at an ultra-high level to provide anti-inflammatory activity to individual animals. The detailed description of the present invention is given. Purification procedure The P k inflammatory composition can be partially purified from whole eggs' yolk or egg white. An example of a preferred partial purification procedure is as follows: 1. Prepare water-soluble from an egg Fractions; 2. ultrafiltration of the water-soluble fractions; 3. separation of fractions by ion or anion exchange chromatography; and fr 4, bioassay of anti-inflammatory activity of the fractions isolated The following is a more detailed description of this procedure: The anti-inflammatory composition printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs can be purified from whole eggs, yolk or egg white. In a preferred embodiment, the composition is From yolk Purified. Lipid fraction is removed from whole eggs or yolk by methods known to those skilled in the art. For example, in the case of spray-dried yolk powder, a solvent (propane, butane or hexane or (Solvent solvent), supercritical CO2, enzymes, etc. to complete degreasing, and in the case of liquid egg yolk, decapidation can be accomplished using the caprylic acid separation method disclosed by Lee (U.S. Patent No. 5,3 67,054). Therefore, conventional methods can be used to directly heat or dissolve egg white in liquid or powder form as described in the examples below. After that, the whole egg, egg yolk or egg white is preferably processed into liquid or powder form and further processed. Water-soluble part 17- This paper size applies to Chinese National Standard (CNS) A4 specifications (210X 297 mm 1221772 A7 B7. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of invention (15) (see examples) Step 2: Water-soluble fractions obtained from whole eggs, yolks, or whites are subjected to ultrafiltration using an ultrafiltration system equipped with a 3,000 molecular I cut-off membrane. Molecules with a molecular weight of about 3,000 Daltons are separated from molecules with a molecular weight below about 100 Daltons. After ultrafiltration, the ultramolecular weight of molecules with molecular weights below about 3,000 Daltons is obtained. The filter is lyophilized, weighed, and ready for bioassay testing and further separation. Step 3: The fraction obtained from less than 3,000 channels of urinary ultrafiltration can be passed through, for example, reverse phase south liquid chromatography The partially pure anti-inflammatory composition was separated and purified. As an alternative or additional strategy, the egg anti-inflammatory composition in ultrafiltration can be reanalyzed by anion exchange DEAE-Sepharose chromatography. In the specifically disclosed embodiment, the preferred ultrafiltration system is performed with Amicon RA1000 (3K MWCO) and DEAE ion exchange layer machine. No, for those who know ‘according to this document’ and what is known to those skilled in the art, they can use equivalent techniques and materials to isolate the composition. Step 4: The anti-inflammatory activity of the composition can be tested by standard bioassay methods to determine the anti-inflammatory activity. Some examples among others include inhibition of white blood cell migration, rat paw edema testing, adjuvant-induced arthritis, collagen-induced arthritis, and life-time microscopy. It is also possible to compare egg anti-inflammatory compositions with known anti-inflammatory guides such as aspirin and indomethacin. Finally, you can use rheumatoid arthritis, degenerative arthritis-18- (Please read the precautions on the back before filling this page) -emmBm n i_l «KKK aMmmB mmmmme j -Kmt IK o— BmmK —Bn- ir -1 ^ This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) 1221772 A7 B7 V. Description of the invention (16) and clinical examination of wounds and arthritis to determine anti-inflammatory activity. The anti-inflammatory effect of some pure egg anti-inflammatory compositions is given by bioassay '/ J. A preferred bioassay is a mouse type 11 gel assay as listed in Example 6. Another bioassay includes pleural leukocyte migration inhibition assays; as contained in the following data: Vinegar et al ,, Some Quantitative Characteristics of Carrageenan Induced Pleurisy in the Rat, `` Proc. Soc. Exp · Bio · Med · 143: 711_714 (1 973), Ammendola, G. et al., &Quot; Leukocyte Migration and Lysozomal Enzymes Release in Rat Carrageenan Pleurisy, &quot; Agents and Actions 5: 250-255 (1975) 'Vinegar, R et al.5 &quot; Printed by the Quantitative Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the notes on the back before filling this page)

Studies of the Pathway to Acute Carrageenan Inflammation, &quot; Fed · Proc. 35: 2447-2456 (1 976) White blood cell movement inhibition test is another useful test, which is usually implemented as follows: a combination of anti-inflammatory A sample of the animal was administered to female rats artificially inflamed with a 1% carrageenan solution, and the decrease in the number of white blood cells contained in the pleural exudates of the treated mice relative to the number of control mice was determined (analyzed by automatic imaging) Technique) The anti-inflammatory effect of each dose of the sample. In addition, the anti-inflammatory effect of a substantially pure anti-inflammatory composition can be tested on the edema caused by carrageenan injection in the meat feet of mice (Winter, CA, Risley, GA Nuss, AW, &quot; Carrageenan-Induced Edema in the Hind Paw of the Rat as an assay for anti-Infammatory drugs, "Proc · Soc · Exper. Biol. -19- This paper size applies to China National Standard (CNS) A4 (210X 297 public broadcasting) b____ Central Standard of the Ministry of Economic Affairs A7 __B7_ printed by Bureau of Consumer Cooperatives Invention Description (17)

Med. 3: 544 (1 967)) ° In addition, a variety of bioassays can be used (see Wetnick, A. S., and S abin, C ·, The Effects of Clonixin and B ethaurethasone on Adjuvant-Induced Arthritis and Experimental Allergic Encephalomyelitis in Rats5M Jap. J. Pharm. 22:74 1 (1 972)) ° Ingredients for partially purified anti-inflammatory compositions. Purified anti-inflammatory compositions have the following characteristics: It has anti-inflammatory activity; ⑵ It is present in the egg white and yolk of avian eggs; ⑶ After isolation from egg yolk, the person isolated from the egg white has greater anti-inflammatory activity; ⑷ has less than about 3,000 channels ear分子量 molecular weight; (0) non-protein and non-steroidal; (6) with thermal stability; 口服 oral effect and will not be broken down by digestive enzymes; 3,000 迢 molecular weight is isolated from the composition part He Purification has introduced an ultrafiltration, membrane in which the separation and purification procedure uses an ultrafiltration, membrane that does not allow greater than 3,000 channels / knife species to penetrate. Partially purified anti-inflammatory compositions have been tested. $ Non-proteinaceous and non-steroidal ones are small in size and cannot be broken down by enzymes that degrade protein I. Furthermore, the composition is orally active and is not degraded by digestive enzymes. A small, stable form of partially purified anti-hair, mouthwash (to distinguish it from larger proteins) helps its absorption from the digestive tract. After removal ’, the partially purified anti-inflammatory composition has thermal stability 1221772 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 V. Description of the invention (18). The Shaofen purified egg-derived anti-inflammatory composition can be isolated from whole eggs, yolk, and egg white. The anti-inflammatory composition obtained from the yolk portion purification showed higher anti-inflammatory activity than the anti-inflammatory composition obtained from the egg whites partial purification. The inventors of the present case have discovered that partially normal purified anti-inflammatory compositions can be isolated from normal eggs and agars from whole eggs produced by hyperimmunized oviparous animals. Hyperimmunization of egg-laying animals As mentioned just now, the inventors of the present case have found that there are ultra-normal levels of anti-inflammatory compositions in hyper-immune eggs that can provide anti-inflammatory activity to individual animals. Immune eggs or egg products can be produced by any species of ovipositor. Preferably, the animal is a member of the Ornithoptera or in other words, a bird. Within the bird class, poultry is preferred, but other members in this class, such as turkeys, crows, and geese, are also suitable sources of hyperimmune egg products. When their oviposition animals are brought to a specific state of immunity through the use of, for example, a periodic booster administration of immunogens, these animals lay their eggs, which have beneficial properties, including A normal level of anti-inflammatory composition that is effective in treating and preventing inflammation-related diseases and autoimmune diseases of the individual animal. Immunosensitivity alone is not sufficient to promote the occurrence of supernormal levels of egg-derived anti-inflammatory compositions in eggs, such as edible eggs that do not contain their super-positive facts, even if the bird is normally immune to bird disease The same is true for sensitization of various immunogens during and during normal exposure to environmental elements. The inventor of this case found that the egg is only in the hyper-immune state of the material. • 21-This paper size applies the Chinese National Standard (CNS) M specification (21〇x 297). (Please read the precautions on the back before filling this page)

1221772 A7 printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. B7 V. Invention Description (19T ^ ^^ &quot;-~~ Only the anti-inflammatory composition with a desirable supernormal level will be obtained. Therefore, the egg will contain a higher level of anti-inflammatory. The particular hyperimmune state of the composition is preferably achieved by administering the initial immunization, followed by periodic strengthening with a sufficiently high dose of a special immunogen or immunogen mixture. The preferred dose of the booster must be equal to or greater than the yield 50% of the dose required for the initial immunization of the bird. Therefore, there is a threshold dose of a booster, below which no such properties can be produced in the eggs of the bird, which means that the bird is in a state generally called immune This is also the case. After knowing the requirements for developing and maintaining a hyperimmune state, those skilled in the art can vary the amount of immunogen applied according to the genera of the oviposition animals used in order to keep the animal in a hyperimmune state. The better hyperimmune state is produced by any immunogen or combination of immunogens. The better hyperimmune state is multiple exposures to multiple immunogens and multiple exposures to a single immunogen Or achieved by a single exposure to an immunogen pool. Almost any immunogen can be used to liberate a hyperimmune state, including, but not limited to, 'bacterial' viruses, protozoa, allergens, fungi, or cellular material. In understanding the development and After the requirement to maintain the hyperimmune state, those skilled in the art can vary the amount of immunogen given according to the genera of the oviposition animals used in order to maintain the animal in the hyperimmune state. In addition to immunization with naturally occurring immunogens In addition, immunogenization can also be accomplished with combinatorial chemistry-derived immunogens. The basic strategy is to create a group of dissimilar molecules by combining multiple movements of chemical building blocks to produce a group of dissimilar molecules. Several species have recently been developed The method is used for the oligomer library (Fodor, S, et al., Science 25 1: 767 (1 99 1); Hughton, R. et al., Nature 3 54:82 (1 99 1)) and small Yes -22- This paper size applies to the Chinese National Standard (CNS) A4 specification (210X 297 Gongchu) — (Please read the precautions on the back before filling this page) Order d ^ ί ΙΒΚ · 1221772 A7 B7 V. Description of the invention (2 ) Organic molecules Bunln, B. &amp; EUman, J, J Am Chem sQe 1 14: 1 0997 (1 992) solid phase and solution phase synthesis. Rapid multiplex peptide and oligomer synthesis can be used as a combination derived immunogen Source. Furthermore, another strategy is to add organic combination elements to the backbone molecule in a combined way to improve the immune system. Other ways to hyperimmunize the product can be used to replace the immunogen vaccine and include the use of gene vaccines. In particular, any DNA construct (usually including a promoter region and an antigen coding region) can initiate an immune response. Gene vaccines include: two antigen-encoding fragments that are radiated to generate an immune response, naked d NA fragments, plastid DNA, DNA-RNA antigens, dna-protein complexes, DNA-liposome complexes, DNA expression libraries, And viruses and bacteria. DNA delivery methods include, among others, particle impingement, direct injection, viral vectors, liposomes and ejectors. When using these delivery methods, much smaller amounts can be required and can often result in more sustained immunogen production. When using their genetic method, the preferred method for introducing DNA into birds is by injecting DNA muscle into the pectoral muscle.

DNA delivery methods include, but are not limited to, particle impingement, direct injection, liposome ’ejector injection (Fynan, E F et al, Proc NaU, printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

Acad. Sen. USA 90: 1 1478- 1 1482 (1 993)). Nucleic acids encoding known or unknown immunogens, promoter genes (noticeable CMV cauliflower mosaic virus) and S V40 bacterial protozoa can be replicated in bacteria to produce plastid DN a for DNA> Wang She. Although several parenteral routes of DNA administration are effective in difficult bodies, the preferred method is intramuscular injection into the pectoral muscles. Vaccine testing is more difficult in spawning birds. During the period of up to 6 months, the standard of China Paper (CNS) A4 is applicable to this paper size --- 1221772 A7 B7 V. Description of the invention (21) Repeated immunization at intervals of two weeks deal with. The preferred amount of DNA is usually in the range of 50 to 300 micrograms of DNA per saline for direct injection. For particle impaction, it is preferred that d n A is co-sedimented to gold beads by adding 4 to 100 micrograms after adding 2.5 M C a C12. This method of accelerating the coated particles with a diameter of D N A into a moving object can be repeated by intradermal immunization. The following is a detailed description of a preferred procedure for bringing the oviposition animal to a highly immune state so that the resulting hyperimmune egg or egg product can be administered to a single animal: 1. Select one or more immunogens. 2. Immune response of spawning animals through primary immunization. 3. Promote a proper dose of immunogen booster vaccine to induce and maintain a hyperimmune state. 4. Check the level of anti-inflammatory activity of hyperimmune eggs. 5. Collect and dispose of the eggs. Below is a more detailed description of this procedure. Step 1: Printed by the Consumer Cooperative of the Central Standards Bureau, Ministry of Economic Affairs. Any immunogen or combination of immunogens can be used as a vaccine. These vaccines can be bacteria, viruses, protozoa, fungus' cells, or any other substance that the immune system of the spawning animal responds to. The key point in this step is that the immunogen must be able to generate immune and hyperimmune states in the egg-laying animal. Although only a single immunogen can be used as a vaccine in the method of the present invention, a preferred vaccine is a multivalent bacterial and viral antigen selected from the following antigen families: Enterobacteria and Bacteroides, Pneumococcus, Pseudomonas, Salmonella, Streptomyces Cocci, Bacillus, Staphylococcus, Neisseria, Clostridium, Mycobacteria, Actinomycetes -24- ^ Paper size applies to China National Standard (CNS) A4 (210X297 mm) '&quot; 122 72 Central Bureau of Standards, Ministry of Economic Affairs A7 printed by employee consumer cooperatives _____B7 ___ V. Description of the invention (22) Bacteria 'Chlamydia' and Mycoplasma. Viral antigens are preferably selected from the following antigen families: adenovirus, picornavirus, and herpes virus, although other viral antigen families can also be used. In a preferred embodiment, a multivalent vaccine called Series 100 (s_100) is used. Bacteria included in the S-100 vaccine are listed in Table 1 of Example 1. Such vaccines have been previously described in U.S. Patent Nos. 5,106,618 and 5,21,576,46, both of which have been assigned to st 0 i j e

Research and Development Corporation. Step 2:

The distant epidemic can be a killed or weakened vaccine and can be administered by any method that can shed an immune response. Preferably, the immunization is performed by intramuscular injection of an immunogen. The preferred muscle for poultry injection is the pectoral muscle. The preferred dose is 0.05-5 mg of immunogenic vaccine. Other methods of administration that can be used include intravenous injection, intraperitoneal injection, intradermal, rectal suppositories, aerosols, oral, topical or ocular administration. When using DNA technology for hyperimmunization procedures, much smaller quantities are required, typically 300 micrograms. Bian Guohui has a variety of methods known to those skilled in immunology to determine whether the vaccine sheds an immune response in the oviposition animal. Examples of these methods include enzyme-linked immunosorbent assay (ELISA), antibody 4 presence test for stimulating antigens, and tests designed to assess the ability of host immune cells to respond to the antigen. The minimum amount of antigen required to defy the immune response will depend on the vaccination procedure used, including the type of adjuvant and antigen formulation used and the type of oviposition animal used as the host. (Please read the notes on the back before filling this page)

-25-

1221772 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (23) Step 3: Those with better hyperimmunity status are defamated in the target animal by repeatedly intensifying the application of the appropriate dose at fixed intervals Keep it; this time interval is preferably between 2 and 8 weeks between 6 and 12 months. However, it is important that booster administration does not lead to immune tolerance. Both methods are well known in the art. Other immunological maintenance procedures or combinations of procedures can be used, for example, intramuscular injection for primary immunization and intravenous injection for booster injection. Other red sequences include simultaneous administration of microencapsulated and liquid antigens, or primary immunization by intramuscular injection, and booster administration via oral or parenteral administration via microencapsulation. Several combinations of primary and hyperimmunization are known to those skilled in the art. Step 4: It is necessary to test the level of anti-inflammatory activity of eggs. This can be done by any clinical and preclinical assessment of the effects of hyperimmune eggs, or derivatives thereof, on inflammation. Inflammation induced by chemical slander in mice is a better standard test for anti-inflammatory drugs (see Example 6). Step 5: This step involves the collection and treatment of eggs containing an anti-inflammatory composition. Eggs can be collected by conventional methods. Egg treatment can be accomplished in a number of ways as described later in this document. Eggs can also be further processed to purify the anti-inflammatory composition described below. Eggs or parts thereof collected from hyperimmune animals, including partially purified -26. This paper is in accordance with the Chinese National Standard (CNS) A4 specification (21〇 &gt; &lt; 297 mm). ) (Please read the precautions on the back before filling this page) Order -n · fj tiH— m A7 ------ ---- B7 V. Description of the invention (24) Anti-inflammatory composition, it is best to Processed to produce a hyperimmune egg spawn that can be subsequently administered to an individual animal. Ie itself or it. Injury body 'includes the partially purified anti-inflammatory composition of the present invention for administration to an individual animal by any means for treating or preventing inflammation, including arthritis, and / or autoimmune disease in an individual animal. Preferably, the egg or any derivative of the egg is directly fed to the individual animal. It is important to note that All India, Yolk, and Egg White are non-toxic and safe natural food ingredients. In another embodiment, the egg or any part thereof, including a partially purified anti-inflammatory composition, is integrated into a nutritional supplement. To be added to the nutritional supplement. A preferred method of preparing the eggs or any part thereof includes scavenging the eggs into egg powder. Although several methods are known for drying eggs, spray drying is still a preferred method. The process of spray drying is known in the art. Their dried egg powders can be incorporated into beverages in the form of, for example, protein powders, energy drinks, insect white supplements and any other nutritional, sports related products. In addition, egg powder can be used in baking mixes, vitality bars, candies, biscuits, etc. Other examples of egg processing include making omelettes, soft-boiled or hard-boiled eggs, baking eggs, or, if needed, raw eggs or processing into liquid eggs. Finally, 'further isolation and purification', as in the case of partially purified anti-inflammatory compositions, can be used for other administration methods such as parenteral, subcutaneous, intravenous' intramuscular, intraperitoneal, intranasal, oral or topical Give egg products to ground. In addition, 'their further separation may facilitate the production of encapsulated products and pharmaceutical compositions using the eggs or parts thereof. -27- This paper size is in accordance with Chinese National Standards (CNs) A4 (210X 297 mm) 1221772 V. Description of the invention (a) Intestine: anti-inflammatory composition preparation package or water-based drug; night, suspension is inferior Examples of alcohols, polyethylene glycols, aqueous solvents or drugs are propionate such as ethyl oleate. For example, in the case of olive oil and injectable organic esters, when the anti-inflammatory composition is packaged, it is preferred to perform oral administration via solid dosage form, human m + pills J, government agents and granules. = _, The anti-inflammatory composition is mixed with at least one inert diluent, such as sugar or dian powder. Their dosage forms may also include, for example, those in the industry, in addition to inert diluents. In the case of capsules, lozenges, and pills, 嗲 箬 Shi 'and Temple shape can also include buffering agents, pH-sensitive salty polymers, or any combination of Pycnogenol and P-sensitive ... Encapsulants are those typically used as encapsulating compositions in the food and pharmaceutical industries. Lozenges and pills can be additionally prepared with enteric coatings. Anti-inflammatory compositions for oral administration Liquids include pharmaceutically acceptable emulsions 'Solution' suspensions, syrups and tentacles, which contain inert diluents commonly used in medical technology. In addition to inert diluents, the composition may also include wetting agents, emulsifying agents, suspending agents, and sweeteners. The Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs printed the hyperimmune eggs or egg products of the present invention, including partially purified anti-inflammatory compositions, which can effectively treat and prevent inflammation, and in particular, all forms of: arthritis, including, but Not limited to rheumatoid arthritis, osteoarthritis, joint adhesion seronegative spondylosis, reactive arthritis, chronic fatigue syndrome, fibromyalgia (fibritis) and gout ^ The printed products of the present invention are the same Can effectively treat autoimmune diseases, such as rheumatoid arthritis, juvenile diabetes' relapsing sclerosis, Gravts's disease, Meneri's disease, myasthenia. 28- This paper standard applies to China National Standard (CNS) A4 (210X 297 mm) A7 B7 Five invention descriptions (26) Weakness' red "Sore itch, systemic scleroderma, wind and SJogren syndrome, etc. (see this document, background" paragraph bu ... Used for the treatment and prevention of special diseases, whether it is general inflammation or immediate inflammation or autoimmune diseases, at the time, the super immune products or any live knife &amp; including partial purification The anti-inflammatory composition, preferably μ 4 ±, can effectively treat and prevent this particular disease to individual animals. # ^ ^. The duration and intensity of the treatment depends on the particular condition, whether it exists or not, and If present, the condition progresses in the individual animal. This hyperimmune egg house or any part thereof, including partially purified anti-inflammatory compositions, can also be used to treat and / or prevent conditions and symptoms < Anywhere | 疋 i ,. For example, in some cases, depending on the particular situation of the situation, individual animals can be administered from less than one full hyperimmune egg to several full hyperimmune imprints (or containing less than one to several Daily doses in the range of full hyperimmune (hyperimmune, i.e. product). The more powerful fractions can be separated and concentrated by the methods described in this article and their techniques known in the art. The egg product of the present invention and the partially purified anti-inflammatory composition were found to be effective in treating rheumatoid arthritis in mice by using an animal model of collagen-induced arthritis (see Example 2). This animal model is It is considered to be equivalent to the effect of rheumatoid arthritis in humans. In addition to the treatment of arthritis, it has also been determined that the egg product can be delayed and given to mice when it is deduced for arthritis, and in some cases, Prevents the onset of arthritis symptoms. Therefore, the egg product of the present invention can not only effectively treat disease symptoms, but also delay and / or prevent the onset or progression of diseases. In another embodiment, the egg product is tested in the human body and proved to be affected by several -29- 1221772 A7 B7 V. Description of the invention (27) Please read the 5th item before filling in the body with signs of arthritis. Positive effects for treating various forms of arthritis (see Example 9). A person treated with the egg product exhibits clinical reductions in symptoms such as pain in addition to the general reduction in swelling and stiffness. The effect on autoimmune diseases is shown by the surprising reduction of the egg product of the invention and the partially purified anti-inflammatory composition against type II collagen antibodies, which is also seen in mice (see the Examples) 3). The printed product of the present invention is expected to effectively reduce antibodies involved in other autoimmune diseases; for example, juvenile diabetes, relapsing sclerosis, Graves' disease, Meneri's disease, myasthenia gravis, lupus erythematosus, psoriasis, systemic crust Disease, rheumatic fever and Sjogren syndrome. As stated in the background of the invention, arthritis patients are currently treated with non-steroidal anti-inflammatory drugs (NS AIDs) and / or disease-modulating anti-arthritis drugs (DMARDs) to resolve the symptoms of arthritis conditions. As shown in the example (see Example 9), the patient can administer the egg product of the present invention simultaneously under the current drug treatment. Egg products work synergistically with medications to reduce the symptoms of specific diseases. So egg products can be administered simultaneously with additional treatments to produce synergistically better results against arthritis and autoimmune diseases. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs In addition, as is well known in the art, NS AIDs and DMARDs can cause severe gastrointestinal injuries. U.S. Patent No. 5,7,72,9,99, assigned to DCV, Inc., and incorporated herein by reference, discloses that hyperimmune egg products can effectively treat gastrointestinal injuries caused by InS AI D-derivation. Therefore, another embodiment is expected to be useful in situations where an individual animal is receiving or preparing to receive NSAIDs to treat an arthritic condition. In their case, the individual animal can also take the egg product of the present invention in order to not only treat the condition of arthritis, but also protect the gastrointestinal system from -30- This paper standard is applicable to the Chinese National Standard (CNS) 8-4 specification (21〇 χ 297 mm 1221772 A7 B7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (28 NS AID p show injury. In their treatment, lower doses of hyperimmune eggs may be sufficient. In addition, patients suffering from Individual animals with this disease can take egg products before they start to administer dual NSAIDs and DMARDs to better prepare the gastrointestinal system. They can be administered / orally by taking the eggs, egg products, and / or anti-inflammatory compositions of the invention Examples of inflammatory conditions treated include acute and subacute mucocystitis, acute non-specific tendinitis, systemic lupus erythematosus, systemic dermatomyositis, acute Fengcheng ^ ^ ^ sky cancer sore 'large cancer dermatitis, herpes stigmatitis , Severe redness, multiple forms of exfoliative dermatitis, sclerosis, seasonal perennial rhinitis, bronchial asthma, atopic dermatitis, serum disease, keratitis, eye iritis, diffuse ureitls, Acoustic inflammation ’optic neuritis, sympathetic ophthalmitis, symptomatic sarcoidosis,

Loeffler's syndrome, chronic pneumonia caused by inhalation of beryllium powder, hemolytic anemia, rheumatitis, mastitis, contact dermatitis, allergic combined meningitis, psoriasis, and joint adhesion spondylitis, Acute gouty arthritis, herpes zoster rheumatic joint 1, osteoarthritis, any other degenerative joint disease, and any other related autoimmune disease. Furthermore, isolated and purified egg products can be used to treat individuals exposed to potentially inflammatory agents such as allergic agents. The effective amount is related to the administration of a hyperimmune egg or egg product to an individual animal. According to the determination and detailed in the following F examples, the preferred dosage range of the hyperimmune egg or egg product to be administered to an individual animal is i. Between milligrams and 10 grams per kilogram of body weight. With regard to the partially purified anti-inflammatory composition itself, the "partially purified composition" has been extracted from whole eggs, whole yolks, egg yolks, and egg whites of pure-31-knife, the preferred dosage The range is from 1 g to 400 mg of anti-inflammatory composition per kilogram of body weight. The dosage of the active ingredient can vary; however, if necessary, the amount of the active ingredient should be adjusted to the appropriate dosage form. It is generally accepted that the formulation chosen will depend on the desired therapeutic effect, route of administration and duration of treatment. The advantageous properties of this month can be observed with reference to the following examples of the present invention. Examples Example 1 One of the Uoo vaccines will be disclosed in U.S. Patent No. 5,2 1 5,746 and contains the detailed list listed in the table, referred to as ', series 100' or 'S_100,' a multivalent vaccine ( From the

Amenean Type Culture Collection) with 15 ml of culture medium and reconstituted at 37 ° C. (Incubate at 3 times throughout the night. After good growth is obtained, inoculate one liter of broth with broth suspension of broth, and incubate at 37 ° C. The remaining suspension is transferred to Sterile diol tube and stored under _ 2OI for up to 6 months. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. After seeing good growth in the culture medium, centrifuge to collect the bacteria. Resuspend the bacterial pellets in the sterile The bacterial sample was centrifuged in a physiological saline solution, and the bacteria were washed three times. After the third washing, the resulting pellet was resuspended in a small amount of double distilled water. The culture-free bacterial suspension was placed in a glass flask and placed in 8 〇The suspension was killed by heating in a water bath overnight. The viability of the broth culture was tested with a small amount of heat-killed bacteria. The heat-killed bacteria were used to inoculate the broth, -32- Standard (CNS) A4 specification (21〇χ 297 mm) 1221772 A7 B7 V. Description of invention (30) Incubate at 37 ° C for 5 days and check its growth every day, because the bacteria must be killed for vaccine use Therefore. Will The heat-killed bacteria are freeze-dried. Then dry bacteria and sterile saline solution are mixed to a concentration of 2 · 2 X 10 08 bacteria cells / ml of saline (丨 660 nm optical density reading). Table 1 S-1 0 0 Antigens in vaccines

Name Medium Name Medium Staphylococcus simulans BHI Pseudomonas aerudnosa, BHI Staphylococcus epidermidis BHI Klebsiella pneumoniae ^ BHI _ Pus # Streptococcus Dvoeenes, Salmonella typhimurium (Salmonella tvphimurium ') BHI brewing pus # Streptococcus Dvoeenes), A Types 3 APT HaemoDhilus influenzae BHI Streptococcus Dvoeenes, A Types 5 APT Streptococcus Streptococcus mitis, APT Streptococcus pyogenes, A Types 8 APT Proteus vulearis BHI Streptococcus Dvoeenes, A Types 12 APT Shigella iShkella dvsenteriae BHI Streptococcus APT pneumococcus (Diplococcus APT pvogenesl A Tvpes 14 Dneumoniae) Streptococcus pyogenesl ATvpes 18 APT iProDionibacter acnes type broth APT Streptococcus saneuis APT Aerobacter £ 〇li BHI 4 way # Streptococcus salivarus APT Escherichia coif BHI Streptocccus mutans BHI Printed by the company (please read the precautions on the back before filling this page) -33- This paper size is applicable to China National Standard (CNS) A4 (210X 297 mm) 1221772 A7 V. Description of Invention (31

Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs Prepare the killed pathogen preparation as described above. For the first vaccination, the bacteria were mixed with complete Freund's adjuvant and 56 grams of bacterial material was injected into the chicken's pectoral muscles. For the rest of the vaccination, the bacterial preparation was mixed with incomplete Freund's adjuvant and injected into chickens at two-week intervals for up to 6 months. . After eggs are collected from hyperimmune chickens, they are spray-dried into powder form. In the spray drying private sequence, the population temperature must not exceed 32 (Γρ, the outlet temperature is maintained according to the generated sum in the range of 3 · 0 · 4 · 0% of the finished product temperature, and the chest pressure system is maintained at about M0 0 to 4000 The PS was monitored at a low temperature of 100-160 ° F and the humidity of the sample was monitored during the drying process to obtain the final product with any desired consistency. Example 2 Purification of anti-inflammatory compositions The example illustrates the method used to obtain a partially pure, low molecular weight, non-aggregated form of anti-inflammation composition from poultry eggs (suitable for large-scale purification). The whole egg two, the immune seal and the control food seal are broken, and the egg white is separated from the yolk The two were spray-dried. Hyperimmune eggs were obtained as described in Example 1. Egg white powder was separately processed to obtain aqueous fractions for ultrafiltration. All purification steps are reducing possible bacteria or pyrogens. Contaminated. The solution is prepared with sterile water and all glassware is depyrogenated. In addition, the solution is sterile-filtered. -34- This paper is sized for CNS) A4 size (2 公 297mm ^ 7 --- -------- ---- IT — ^ ----- # (Please read the notes on the back before filling in this page) 1221772 Employees' cooperation cooperation of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (32) Example 2 a Solvent extraction of egg-derived anti-inflammatory composition from egg yolk powder The dried egg yolk prepared according to Example 1 was subjected to liquid solvent extraction with propane or butane to separate lipids from the aqueous egg yolk portion containing the anti-inflammatory composition. In other words, 500 g of dried egg yolk powder was placed in a tube of tincture, and 4 liters of liquid propane solvent was added thereto. The solvent solution and the extracted lipids were taken out. Six additional solvent extractions were performed to obtain a total of six lipid extractions. Ultrafiltration dilutes 400 grams of dry defatted yolk with 4 liters of sterile steamed water and homogenizes with virtis (handishear). The yolk mixture is centrifuged at 24 RPM or left to stand until undissolved yolk particles precipitate. Ultrafiltration was performed using an Ami con RA 1 000 ultrafiltration system equipped with a spiral wound membrane with a cutoff rate of 3,000 channels. The pump speed was maintained at 20 ° with inlet pressure and I5 psi outlet pressure. 0.45 microns was used. Sterile Disposable Nalgene The sterilizer aseptically filters &lt; 3,000 molecular weight permeate, and freezes it to dry or freeze it for storage, bioassay or further purification. Figure 1 shows the flow chart of the purification procedure. The molecular species of less than 3,000 canals have anti-inflammatory compositions in a low molecular weight non-aggregated form. Approximately 12 grams or 3% of the entire anti-inflammatory composition is obtained from 400 grams of starting material. This 3 KDa portion was tested for anti-inflammatory activity by bioassay and showed a high level of activity (Example 5). Example 2 b Preparation of egg-derived anti-inflammatory composition from blanket egg white powder -35- This paper size applies Chinese national standards ( CNS) A4 size (210X29? Mm) (Please read the notes on the back before filling out this page) -1%. Order 1221772

Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. The Choke White isolated from the hyperimmune eggs and control edible eggs described in m was diluted with 4 liters of deionized water. The mixture was decanted and filtered through a -40 micron scaler and ultra-filtered through a 3 KDa Mwco ultrafiltration system (Figure 2). From the grams of egg white powder, 8.6 grams or 2.15% of the anti-inflammatory composition was recovered. This example shows the preparation of an active anti-inflammatory composition from egg white matter of both hyperimmune eggs and control edible eggs. Example 3 E ion exchange layer folding The egg-derived anti-inflammatory composition was further analyzed by anion exchange DEAE chromatography.

In the same manner as described in Example 2, J, 000-channel ear canal ultrafiltration penetrating fluid was prepared from both defatted yolk and egg white. A DEAE Sepharose column (2.5x40.0 cm) was pre-treated at room temperature with 1 000 ml of 20 mM, pH

8.0 'Acetic acid in pyrogen-free water in a sterile double-distillation chamber was equilibrated with buffer. Dissolve defatted egg yolk (15 g) or white (18 g) 3,0 ear canal penetrating solution in 200 ml of sterile pyrogen-free double distilled water and apply it at a flow rate of 00 ml / hour DEAE Sepharose column (Pharmacia Biotech., Fast flow). The components contained in the 3,000 ear piercing solution were eluted with the following buffers to separate the egg-derived anti-inflammatory composition and its molecules. At the same flow rate, DEAE Sepharose was eluted with 200 ml of 20 μM pΗ 8.0 ammonium acetate and 200 ml of 20 mM ammonium acetate pH 6.5 to remove unbound material. Then, 500 ml of 150 m M p Η 5.0 acetic acid was applied to elute the weakly bound molecules. The stronger binding molecule was washed with 200 ml of 150 m mM ammonium acetate pH 4.0. Finally, use 400ml 1.5N NaCl / 20 mM 36. The paper size is applicable to Chinese National Standard (CNS) A4 (210 X 297 mm) (Please read the precautions on the back before filling this page) Order 1221772 A7 B7 V. Invention Description (34 Ammonium acetate, pH 6.5 regeneration of DEAE Sepharose column (Figure 3). Figure 3 does not show a typical chromatogram of 3 Kda penetrating fluid obtained from defatted yolk and egg white using DEAE Sepharose. Letters indicate the use of buffers The obtained elution pattern of the liquid was collected. Each part of the 15 ml of the eluate was collected and the molecular absorbances at a total of 2800 nm and 220 nm were measured. The 4 eluate fractions were combined and lyophilized to remove water. Table 2 The separation results are summarized. Table 2 DEAE Separation Carcinoma Volume (ml) of 3,000 Ear Penetrating Fluids of Defatted Yolk and Egg White Weight of Extraction Well (g) Recovery Rate Please read the back 5 first and fill in this Page 1-22 23-40 41-56 57-70 2-26 27-46 47-60 61-73 294 252 224 196 350 280 196 182 11.79 0.40 0.44 0.19 15.33 0.23 0.32 0.44 78.6 2.7 2.9 1.3 85.2 1.3 1.8 2.4 Examples 4 3,000 Ear Ear Penetrations

Liquid and its DEAE

Sepharose Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. 逆 PLC analysis liquid chromatography (Η PLC) (S himadzu Unit, SPD-M 10A VP Diode array detector, LC -6AD Liquid Chromatograph's SCL-10A VP Degasser, LPM-600 Low-pressure Mixer, and CTO-10A VP Column Oven) Analyze 3, 000 ear piercing fluids from yolk. All eluent fractions were prepared as 5.0 mg / ml ultrapure water and then filtered through a 0.2 micron filter. For example, 20 mechanical liters of each sample is applied at 29 C Waters Symmetry -37- This paper size is applicable to China National Standard (CNS) A4 specifications (210X 297 mm) 1221772 A7 B7 2. Description of the invention (35 C18 column (3.9x150 ^ m). The flow rate is 10 ml / min. A linear gradient of 0% -60% acetonitrile / d H20 / 0.1% TFA is used as the mobile phase. Figure 4 shows Max plotting chromatogram of Max produced by defatted PL-100 egg yolk with 3,000 channels. The same analysis conditions were also used for 3,000,000 obtained from defatted edible egg yolk. The chromatographic results were obtained from the Dalbergia penetrating solution (chromatograms are not shown). Table 3 summarizes the HPLC analysis results of 3,000 Dalbergia penetrating solutions. The Qin solution with a peak area greater than 0.5% 1 The fractions were collected. The similarity of the spectrum maggots was compared within their retention time +/- 0.5 variability. The common spectrum peak means that they exist in both PL-100 yolk and edible egg yolk? 1] 〇〇 The band indicates that it is only present in PL-100 eggs. Comparing the two parts, p [· 丨 〇〇3,0 0 0 The ear penetrating fluid showed 5 more bands than those of yolk yolk, and it can be speculated that the chicken's hyperimmunization can produce more biological components in its eggs. JKD penetrating fluid common spectrum peak PL_100 spectrum Xiang Stone egg spectrum Qin ~ PL-100 Egg 18 15 — &quot; 〇 ^ Edible egg 18 〇5 23

Preparative HDLC In order to obtain a large amount of anti-inflammatory agents after ultrafiltration, the preferred step is to separate and purify a combination of molecular weights below about 3,000 channels on a preparative PLC column. Thing. Therefore, one was filled with Z rb ax C 8 packing material: &gt;. 0 8 X 2 0 cm tube column and purified in each stream * g ultrafiltration of less than 3,000 channel MW. Use 80/20 H2O / methanol plus 5% 5% TFA as the shift-38- This paper's standard is applicable to the Chinese National Standard (CNS) M specification (2 丨 〇 &lt; 297 mm) ------- -4! (Please read the notes on the back before filling out this page} Printed invention description (36) printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Fraud Economics, the phase is 9G milliN / min. The purified composition was collected and lyophilized. The person who collected the samples then re-collected the peaks in the analytical g Wang Yi β. In the method described above, the activity was determined by preparative HPI ^ C and analysis and analysis. Elemental composition and chemical structure, a large number of ultrapure spectra were obtained. 7 Mass spectrometry, elemental analysis, infrared spectroscopy, and nuclear magnetic resonance (NMR) were used to determine the structure of the factors. Sex-Quality &quot; _ &quot; Lao ^ In vivo collagen and arthritis is a universal experimental animal model of rheumatoid arthritis and has been used by several practical laboratories and research institutions since 1977. Since its initial discovery, collagen Animal model of induced inflammation has been shown to be related to human rheumatoid arthritis Similarities, for example, and colleagues have shown that there are several clinical signs and changes in medical records in the joints of Guanyinyan mice similar to those of human rheumatoid arthritis (stuan et al, 198 3), especially important for many rheumatoid joints The fact that patients with inflammation show autoimmune to collagen type. In the past, this animal model has proven to be a new inhibitory form &lt; form and a potential immunomodulatory substance in terms of anti-arthritis effect A useful predictive model for clinical benefit. Arthritis in mice and mice is usually induced by the immunization with heterologous type II collagen, which is a response to the initial combined humoral and cellular immune responses of joint tissues. Existing therapies are incompetent or have side effects that limit their long-term use. This example focuses on the egg product and partially purified anti-inflammatory composition of the invention before orally administered to mice before and after arthritic slurries with collagen II. Effective with ^ (Please read the precautions on the back before filling this page) T ,! -39- 122177?

should. The suppression of arthritis was investigated in doses relative to the control group. 30 Sprague-Dawiey rats (Charles RiVers, Wllmington, μα) weighing 100 125 g were randomly assigned into 3 groups (animals / groups). The shellfish line was repeated three times so that the final group included 3 mice. / Treatment resulted in a total of 90 mice in this study. Mouth mist-dried hyperimmune egg products (made according to Example 1) were diluted for oral gavage. Every 10 days, 10% and 0.2% egg product solutions were prepared for oral use. The next day, the solution was stored at 4 C until needed. The egg product (3.5 ml of each solution) was orally fed to the mice seven days before the start of type π collagen and arthritis in the mice and 4 days after the inflammation. The method for inducing and assessing arthritis in mice is based on T r enth am e t al · 1 977. In summary, Spragu and Dawley mice were immunized by intradermal inoculation with 400 hydrocarbon pepsin-treated natural skin] [type 1 chicken collagen (CII) (Genzyme, Boston, MA) and the collagen (cn ) Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling this page). It falls in 0.1 M acetic acid to 1 mg / ml and is not completely treated with the following formula: 1 emulsifier. Three groups: water feeding group (control group), 50 × hyperimmune egg product group (south dose group) (3.5 ml of 10% hyperimmune egg product solution) and 1 × hyperimmune egg product group (low Dose group) (3.5 ml 0.2% hyperimmune egg product solution) 'mother group 10 pairs of mice. The erythema (0-4 +) and rat paw around the paw joints of each limb of the mouse (blind) were clinically evaluated starting on the 10th day after the subcutaneous collagen 11 was immunized and daily until the 21st day. Swelling (0-4 +). The group arthritis index (ΑΙ) is based on the claw score based on the occurrence of arthritis and -40- This paper size applies the Chinese National Standard (CNS) Α4 specification (210 × 297 mm)-printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs装 1221772 V. Description of the invention (38) I is the sum of the severity of arthritis of Am. On day 21, rescued and bleeded. Serum was evaluated against the collagen antibody titer. The ELISA-linked immunoassay (ELISAis) measures physical price against type π collagen ㈣ (Trentham et al. 1 983). Animals were then killed in an overdose: metafane. Results The anti-arthritis properties of DCV egg products were considered in a dose-dependent manner in the collagen type Ⅰ mouse model. "High-dose DCV egg product consumption is clearly demonstrated in this model via the cell and humoral segments that affect the immune system. Provides anti-inflammatory properties. Child compounds appear to be non-toxic at both low and high doses. Compared with the water control group, the high-dose group not only showed a statistically significant reduction in the occurrence of arthritis, but also delayed the onset of arthritis and reduced the severity of arthritis. Figure 5 shows that taking high-dose egg products before immunization with type I collagen can prevent more animals from developing arthritis symptoms without eggs. = In addition, within 11 days of treatment with printed products and later in this Throughout the study period, a significant reduction in the incidence of arthritis relative to the combination that did not receive any eggs was observed. S-100 and PL-100 whole eggs can be used in the first 1] (type collagen deflated arthritis model) Meso-sacrifice shows the effect of sacroiliitis. After the eggs are separated, the yolk shows anti-arthritis activity in the animal model but not the egg white. However, the same yolk is removed and the anti-arthritis activity is eliminated. This This defatted yolk was further purified by a 3,000-molecule cut-off size exclusion column and tested in a rat collagen type 丨 丨 model. The pure fractions of children's liver showed the same results as those shown in the full PL _ 丨 〇〇 eggs.者 相 相 -41-This paper size applies to China National Standard (CNS) A4 (2i × 297mm) 1 «· _», 1.1 _: 丨 i · "! (Please read the notes on the back before filling this page ) ....... fyi-... ί, -mouth ί 1-ί Γϋ 1221772 A7 B7 V. Explanation of the invention (39 South Anti-arthritis activity. Another important observation completed is that mice fed with PLd 00 eggs have antibodies against type II collagen in their serum. The animals in the comparison group were statistically reduced. This point shows that PL-100 eggs have an immunosuppressive response in these animals and therefore help to slow down the arthritis symptoms in their joints. Table 4 Yegao joints The average number of days of the experimental rate of inflammation. Experiment 2 Control group 14 High dose (please read the precautions on the back before filling this page) d Printed by the consumer cooperation of the Central Standards Bureau of the Ministry of Economic Affairs ------ A ^ ____ 1 ^ * The delay in the occurrence of maximal inflammation after immunization with collagen II was delayed on day 2. In animals receiving high-dose hyperimmune products, the arthritis index was reflected relative to the severity of arthritis index. There was a significant reduction in the Zhao group (2.46 ± 0.55 vs. 4 167 soil 〇51; po 〇03, S ^ dent t test). The effect of low-dose egg products was not significantly different from the control group. In addition,相 The incidence of arthritis in the high-dose group It has a significant depression on Zhao New (30% vs. 7G% W3). The reduction in the percentage of arthritis seen with high-dose ultra-free squatters is similar to that of steroid NSAIDS (non-steroids) Anti-inflammatory drugs) methotrexate, as good as those reported by diamine tetracycline.: The examples also show that giving a high dose of egg product before inducing arthritis can prevent more weight loss than those without egg recipients. In Figure 6. Triggered off ... These knotty fingers 42-1221772, Wu Jieming instructions (40 Figure 6 shows the high-dose product to prevent arthritis. Oral gavage animals had a lower percentage of syphilis than control animals that did not receive eggs. Movables were given by oral gavage for 2 i days, seven days before immunization (arthritis) and 14 days after immunization. CONCLUSION: It is shown that egg products have a clear anti-arthritis effect in the type II collagen-induced mouse arthritis model. A statistically significant decrease in the incidence of arthritis was observed in animals receiving high-dose superimmuno ^^ products, with a delayed onset and reduced severity. This data shows that high doses of M have a stinging effect on arthritis. Most importantly, the hyperimmune egg products were non-toxic in the presence of both high and low doses of I. Example 6: Reduction of Libo11 antibody The old man of Example 5 was bled on day 21 and a serum sample was collected. Anti-collagen test using standard ELIS Α test! The potency of autoimmune antibodies. These results demonstrate that animals consuming high- or low-dose egg products can significantly reduce the collagen 11 antibody titer relative to the control group (Table 5). --------- # | 丨 (Please read the notes on the back and save this page)

T Printed by the Consumers' Cooperative of the Central Standard Bureau of the Ministry of Economic Affairs of the control group Low-dose group High-dose group Collagen II antibody potency (-log2) 9 · 3 ± 〇 · 〇3 8.5 ± 0.28 8.3 ± 0.36 Statistical significance — P &lt; 0.004 _ Ρ &lt; 0.005 The potency of autoimmune antibodies against collagen ZZ contained in the serum of animals fed with egg products is higher than that in control animals (without edible egg products) -43 This paper applies Chinese national standards (CNS) A4 specification (210X 297 mm) 1221772 A7 B7 V. Invention description (41 The fact that the printing of employees' cooperatives in the Central Standards Bureau of the Ministry of Economic Affairs has been reduced is of great significance. Because all groups of animals receive the same level of collagen II. The fact that the antibodies against collagen and the resulting arthritis symptoms led to the receipt of egg products on the second day has a lower collagen 11 antibody value is unexpected. This collagen π antibody The reduction of the force value indicates that the hyperimmune egg product has a positive effect on both the cellular and humoral immune responses. Example 7 This example shows that the hyperimmune egg (made according to the example 丨) has a significant effect on food consumption. The anti-inflammatory effect of hyper-immune eggs in dogs with carrageenan on skin edema is confirmed by their results. Their results confirm that hyper-immune eggs can reduce inflammation. Their effects on inflammation can be compared with non-steroid anti-inflammatory drugs. Gbuprofen) was used in this test at a dose of 10 mg / kg, ed ~, and 20 Whlte Eagle Beagles (four groups of 5 dogs each) were fed with black lamb and dogs Rice basic food (35G grams). As shown in Table 2, two dogs (groups i and 2) each had only two basic foods during the conditioning period of about 100 days. In addition, the 2nd The dogs in the group were treated with a nonsteroidal anti-inflammatory drug, ibuprofen, before being stimulated with carrageenan. In the same period, the remaining two groups (3 &amp; 4) were treated with basic food plus super Immunized eggs (HIE) were fed with silver. Group 3 received hyperimmunization with gram added to the food. * Group 4 received τ 3) grams of hyperimmunized mark added to the food. At the end of the conditioning food treatment, it was injected subcutaneously. 2% carrageenan stimulates dogs and induces an inflammatory response. The stimulation procedure is described below. Every day randomly selected from each group—dogs (please read the notes on the back before filling this page) 1r • —--ill I -I m — I i · -44- ~ 1/72

A7 B7 Invention Note (42)-Order Stimulation and Inspection. This procedure was repeated daily for a total of days until all dogs in all groups were tested. 15 minutes before the stimulation procedure, the 2nd, 2nd, and 3rd dogs were orally given 10 mg / kg of ibuprofen. All dogs were anesthetized by intravenous injection and the hair on their left side was shaved (approximately 6 X 8). Add 3 lines and 3 marks to the shaved area and number them! _ Yu Xiong No. 1 was intradermally injected with 0.1 ml of saline as a negative control. : In Uji No. 2-6, intradermal injection of 0.1% halo of 2% carrageenan gel was hardened. The injection was performed by the same person throughout the duration of the study. Intradermal injection of carrageenan at this concentration resulted in measurable swelling in the animal body of 1000%, and their swelling can be reduced with 10 mg / kg of ibuprofen. After the injection of carrageenan was completed, each swelling was measured using a levometer and the value was recorded. Allow the anesthetized animal to recover and repeat the measurement and recording after 6 hours. The average size of the inflammatory response measured for each group--ϊ im «—.— 1 i tm —nil ϋϋ · ml m mu ϋϋ ml ml 1N (Please read the precautions on the back before filling this page) Central Standards of the Ministry of Economic Affairs Bureau Cooperative Consumer Co-operative Prints Sheet 6

These results show that the use of hyperimmune egg silver nougat and basic foods-45- This paper size is applicable to Chinese National Standard (CNS) A4 (210X 297 mm) 1221772 A7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs It shows that (43 grams plus basic food) animals have a significant reduction in swelling, showing that the super: pupa eggs have a preventive effect on inflammation when fed orally. Furthermore, these results show that the anti-inflammatory activity of hyperimmune imprinting can be comparable to that of anti-inflammatory drugs, isobutyl benzoate, and 5 ′. A difference at the significance level of P &lt; 0.0 5 can be seen between the treatment group and the control group. Example 8 A nutritional drink supplement made by Ying Xiangnian, a man who has gone east, includes high-protein, high-sugar powder, which has 302, a recommended daily allowable amount of vitamins, and about 45 grams of super Immune to 17 (about 0.4 eggs). The beverage supplement is made of DCV, Inc,

Wilmington, provided. There are three patients taking this nutritional supplement—a period of two months. The evaluation of the material's t resistance was evaluated by each patient's assessment of clinical status and detailed clinical tests before the study for one month, two months, and one month (one month after the product was discontinued). In addition, clinical chemistry, hematology and urinalysis monitoring are also implemented. The treatment results for each patient are described below: Patient # 1 ° * Patient # 1 is a 29-year-old woman with 5, 3, and 170 pounds of rheumatoid arthritis (naive) medical history. At the first examination, the patient presented with pain, swelling, and tenderness in the joints of the fingers, wrists, toes, feet, and knees. She complained of enlightenment restrictions such as not being able to bend knees or running and having difficulty squatting. Its hematological test has a normal red blood cell deposition rate (ESR) and. Reactive egg (CRP) reading. The patient's cholesterol was 99 g / g. Its slimming use Xie in 3 00 mg i week 'LoeminFe Mm has been treated and -46 This paper size applies Chinese National Standard (CNS) A4 specifications (210X 297 meals) MW-(Please read the precautions on the back before filling (This page) 1Γ 1221772 A7

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. Taking multivitamins: Its diagnosis is rheumatoid arthritis with tis. After taking nutritional beverage supplements on the mother's day for one month, the swelling and swelling were reduced, and the pain and swelling on his feet disappeared. It felt "less painful, 1" and 75% better wealth. The report: There is increased energy. Its ESR and CRp #, / ..., decreased to 186 Gugu / v. The value of Gu remained normal but its cholesterol value was ^ j. The patient continued her medication with an egg-based drink. During the second month, the patient felt her hands were bad and she showed finger spots. She had swollen wrists and reported that her hands were numb at night. Her chat feels “more ill than the one that gave birth to a child ’s life. It ’s estimated that the child is a little worse. She still feels pain when carrying shopping bags or opening the box. Her job title and reading value remain upright and her Cholesterol value remained at 189 mg / gong. It continued to take medicine and stopped using the product according to the schedule of the program. After one month of different products, hands and feet returned to swell and softness. Its functional limit kept its first appearance The patient still feels "% better". The readings of E S R and C R P remained normal while their cholesterol values decreased to 180 mg 1 mol. The score for arthritis symptoms is based on the patient's own assessment of the symptoms. In particular, the patient is asked to indicate the amount of swelling, pain, and / or softness of the joint. Based on this patient evaluation, a value showing the degree of symptoms is generated. The value ranges from a high value of 36 (maximum swelling, pain, and softness) to a low value of 0 (no swelling, pain, or softness). The score from Patient # 1's arthritis condition is as follows: At presentation = 14 —1--I m

fn f—1 g-I e Please read the notes on the back before filling in this page) I- — In i ------ · Order 47- This paper size applies to China National Standard (CNS) A4 (210X 297 (Mm) 21221772 V. Description of the invention (45) Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs for consumption of eggs-based beverages for one month = consumption of eggs-based beverages for two months = one month after discontinuation of beverages # 2 / = 3: 88 women, 5, 3, 1 years old, have a history of rheumatoid arthritis. She showed pain in her lower limbs and shoulders. Physical examination revealed swelling, pain, and softness of the fingers and tread. Functionally it only has a finite square path and it is difficult to bend its arms. The hematological test showed a total cholesterol value of Nannen (36 Amy / hour), CRP (1.64 mg / gong) and 22 mg / gong. His treatment was: methotrexate 5 mg / week, Orudis KT 2 days ’Ca 1 500 mg, Fosomax 10 g, prednisone 5¾ g q2d (every other day), and taking multivitamins. Similarly, people continue to receive these medicines plus egg-based beverages. μ One month after starting her egg-based drink, she still complained of swelling in her hands, pain in her knees, and inability to swell her eyes. The loyalty reported that, 50% is better, "feel better in general" and "go better". He still feels pain, but his pain is less frequent. Hematological results were ESR (30 mm / hour) and cRp (i㈠Pg / Kong) 'and an increase in cholesterol was observed to 2 3 2 mg / Kong. Two months after consuming the product, patients reported that it felt 98% better, but the limits remained the same. Hematological results showed a decrease in e $ R (21 * m / h) and returned to the normal CRp reading (&lt; 0.05 mg / kg), and its total cholesterol remained at 234 mg / kg. The patient continued to take medicine and discontinued the egg-based drink according to the schedule. One month after the product was discontinued, the patient complained of severe joint pain in the fingers and legs. 8 (and please read the precautions on the back before filling this page)-·

IT -48

This paper scale Lai Jin Guanjia ^ 7 ^ 7 ^^ 7 2T〇i29 ^ F 1221772 Α7 Β7 V. Description of the invention (46) Shows swelling of the wrist. It feels that it is &quot; 50% worse. Its ESr (27 mm / hr) level climbed with cholesterol (275 mg / kg), but its CRP (&lt; 0.5 mg / kg) level remained normal. The score for arthritis symptoms is based on the patient's own assessment of the symptoms. In particular, patients are asked to indicate swelling, pain and / or softness of the joints. Based on this patient evaluation, a value showing the degree of symptoms is produced. The value ranges from a threshold value of 36 (the maximum amount of swelling, pain, and softness) to a property of 0 (no swelling, pain, or softness). The score from Patient # 2's arthritis condition is as follows: At presentation = j 〇 One month of consumption of egg-based beverages = 7 Two months of consumption of egg-based beverages = 2 One month after withdrawal of beverages = 1〇 Patient # 3 心 者 # 3 is a 79-year-old man with 51,152 pounds and has a history of osteoarthritis and atherosclerotic cardiomyopathy. The symptoms are pain in the lower back, shoulders, knees and feet. Its medical treatments include: NitrQderm patch, called 譲 0.125, aspirin 80 耄 g and ... to 20 mg. Functional restrictions Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page),-Mouth: It is difficult to move the arm and exhaustion. Hematological results were normal ESR and C R P and cholesterol readings were 2 mg / g. At Drinking # Product-month plus the aforementioned medical treatment_month, the patient felt "slightly better (10.2%)". He still reports right and left shoulder pain and reports that he cannot stand and his right knee is inflated. Functional restrictions remain as reported previously. Its ESR and CRP readings were normal and its cholesterol level was decigrams / mg-49-1221772 A7 B7. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (47) 〇 Two months after eating the product, the patient “Sustained feeling” is slightly better. Symptoms include pain in the right knee and difficulty walking. Cholesterol hard-to-read values were not obtained but E s R and CRP remained at normal levels. After one month without using the product, the patient overall felt slightly better but better than the previous one. Months are slightly worse. Symptoms include swelling of the right leg, shoulder pain, and lower back pain. Joint evaluation indicates pain in the shoulder and lower back. Functional limitation is difficulty in moving arms and feeling, failure, ”ESR and CUP readings are normal and cholesterol Maintained at 18 mg / gong. The score for arthritis symptoms is based on the patient's own assessment of the symptoms. In particular, patients are asked to indicate the amount of swelling, pain and / or softness of the joints. Based on such patient evaluation, A numerical value showing the degree of symptoms is generated. The value $ has ranged from a threshold value of 36 (the maximum amount of swelling, pain, and softness) to a low value of 0 (no swelling, pain, or softness). The score from the arthritis condition of patient # 3 is as follows: At the time of presentation = 11.5 Consumption of egg-based beverages for one month = 2 Consumption of egg-based beverages for two months = 3 One month after discontinuation of beverages = 8 5 Its medical records show similar improvements and confirm the effectiveness of the present invention in eliminating pain and inflammation of arthritic joints without side effects. The present invention has been described with particular reference to certain soft examples, but Some changes and modifications of the reader can be completed within the meaning and scope of the present invention which is sufficient in the scope of the above-mentioned and attached patent applications. -50- This paper Langchi Lincai Guanjia County (CN7TA4 ^ (210X297 ^ 蜷 — 丨 (Please Read the precautions on the back before filling out this page}-Order 1221772 A7 B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of Invention (48) Example 9 Sialic acid release, purification, and analysis of sialic acid Figure

The release and purification of sialic acid from glycoproteins is at a relatively low level of 0-acetamidine. The sialic acid release is sealed with 1 ml of 2M acetic acid in a vial with a crimped top at 80 ° C for 3 hours. Achieved. KDN was added as an internal standard before hydrolysis. After acid hydrolysis, the reaction mixture was cooled and the vial was centrifuged to remove any insolubles. Transfer the supernatant to a mini 5 ml), Centricon (2 ml), or Centriplus (15 ml) 10,000 molecular weight cutoff centrifuge device. The acetic acid was removed from the filtrate by vacuum centrifugation and the sample port σ was replied to 0.5 liter of water. Samples were applied to 1 ml of Dowex 50AG1 X 8 (hydrogen form) / water pipe plug to remove cationic impurities. Collect the flow-through and 4-5 ml of water washing solution in a thin-walled centrifuge tube containing 40 liters of 1 M formic acid. The acidified wash solution should be centrifuged to dryness to remove the weak acid produced by the column.

The sample was re-dissolved in 0.5 ml of 10 μM sodium formate, pH Η 5.50 and applied to 1 ml of Dowex AG 3 X 4 (phosphonate form) weak anion exchange resin buffered with 10 mM sodium formate pH 5.50. Immediately use 5 ml of 10mM -51-This paper size applies to China National Standard (CNS) A4 (21 OX 297 mm) --------- Φ | 丨 (Please read the precautions on the back first (Fill in this page again) Order 1217772 Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs A7 ---------- __ _B7 V. Description of the Invention (49) A branch of elution. Discard the flow-through lotion. Use 10 ml of 1 M formic acid to draw the "dip: acid wash into a thin-walled centrifuge tube and dry the eluent before resuspending in water. Note: For all steps in this procedure, the total amount of ethyl acetate is only-consider Shi 'can be carried out at room temperature. If it is necessary to prevent the removal of β-consumption, the user must use i M acetate acetate buffer at rc, ρΗ 5 5 to replace Xia M for subsequent application. Column description

Dowex 50 AG 1X8, hydrogen form: Take 2 milliliters of resin with a spoon—small, disposable plastic sintered tubing. The column was firstly charged with 5 ml of MH C1, and then washed with 5 liters of water to mention that ρ Η rose above _4.

Dowes AG 3 × 4 (or 4 × 4), formate: Take 1 ml of resin with a spoon into a small disposable plastic sintered tubing column, and convert it to hydroxide ion form by washing the column with M NaOH. ㈣ Use 3_5 ml! M formic acid washes the column or until the pH of the effluent is _ i 5 to convert the hydroxide ion form to the formate form. Wash the column with deionized water. The column was then buffered with 10 m of formic acid steel 'pΗ5 · 5. Dialex HPAEC / PAD was used to analyze sialic acid on Carbopac ρΑιιιη column with Dlonex GP40 gradient pump, ED40 electrochemical detector and CarboPac 1> 8-10 protection column and analysis column with HpAEc / pAD Analysis of sialic acid samples. Sample injection was performed using an AS 3500 autosampler. 100 mM NaOH (eluent 0 and 100 mM NaOH, 1M NaAc (eluent D)) was used to elute sialic acid in a gradient method. -52- used in this method This paper standard applies to Chinese national standards (CNS) A4 specifications (4! Γ Jing first read the notes on the back and then fill out this page "-T 二 -ά 1221772 A7 V. Description of the invention (50) The separation gradient is: time (minutes) A% B% C% D% INJECT TITL1 Start-93.0 7.0 0--93.0 7.0 XX 10--70.0 30.0 11--70.0 30.0 12-93.0 7.0 27-93.0 7.0 fnn 0H an ffi n ft— I -n (Please Please read the notes on the back and fill in this page.) This method is printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs at a flow rate of 1 ml / min and takes 27 minutes. Sialic acid is quantitatively analyzed by using KDN as the internal standard. The area of the sample spectrum was normalized to adjust the loss of KDN signal due to hydrolysis or ion exchange cleaning. Conclusion Analysis of sialic acid ----- ~ free egg component + bound sialic acid __ content / g Egg _% sialic acid yolk yolk 1.9 mg __________. 19% _ edible egg 1 mg. 10% 3k part of PL-100 egg yolk 7.7 mg. 77% _ PL-100 egg white 3k part 铷 0.75 mg. 075% ^ The above table does not show the use of Standard Dion ex protocol and PAD detection Analyze the free and bound sialic acid content of the eggs. This procedure is implemented in conjunction with the preliminary removal of Dowex resin, which results in the full recovery and elimination of other impurities in the chromatography 1. See!- 53- This paper is suitable for financial management. V Name 7 11 2 12 A7 B7 V. Description of the invention (51) The separation of the 3 k parts contained in the egg yolk will not significantly increase the content of ring-bound and free sialic acid The biological activity of yolk 3K in rat type II collagen test cannot be attributed to the sialic acid content due to its very low content (2.7 mg / dose). (Please read the precautions on the back before filling out this Page) Printed by the Consumer Cooperatives of the Central Bureau of Standards and Quarantine of the Ministry of Economic Affairs-54- This paper size applies to the Chinese National Standard (CNS) A4 (210X297 mm) j * &gt; 1-, \--| 1 Issue 86.09.08 Case No. 086112945 Category Hidden 1KK — (The above columns are to be filled by this Office ) A4 C4 Chinese manual replacement sheets (93 January) Cloud patent specification further portion of the purified form of chicken eggs 1,221,772 described anti-inflammatory compositions -, English name of the new invention

CHICKEN EGG ANTI-INFLAMMATORY COMPOSITION IN PARTIALLY PURIFIED FORM Name Nationality 1. Sandra G. Fitzpatrick-Merritte 2. Jeffrey G. Hencha 3. Yang-Zhou Li 4. Lee R. Baker Name of residence and residence (name) Nationality 1.2.4. United States 3. Canada 1. Guiding State, 27 Changjiao Lane, Rose Valley City, Pennsylvania 2 _ 922 Coby Road, West Chester, State of Pennsylvania 3. _ State Ohio No. 9043 Wenlop Road, Cincinnati, USA 4. DCV Bio Inc., No. 1670 North Liberty-Kut Road, Ribano, Ohio, U.S. III. Applicant's Residence, Residence (Office) Name of Representative O : \ 49 \ 49495-4 Oriidas Tyson, 3521 Seaville Road, Wellington, Delaware, USA

930120.D〇aTSP This paper size is applicable to China National Standard (CNS) A4 specification (210X297 public director)

Claims (1)

Case 4 of 3 years (9, please apply for this application for Lisheng special range No. 6. Scope of patent application A B c D L An anti-inflammatory composition in a partially purified form, wherein the anti-inflammatory composition includes the following characteristics: a) it is present in whole eggs and yolks of eggs laid by chickens; b) it is water of hyperimmune eggs Osmotic and fat-removing fraction, its molecular weight is less than 3,000 Daltons; c) it has thermal stability; d) it has oral activity; and e) it can resist the degradation of digestive enzymes; The composition may be administered parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, intranasally, orally or topically; and wherein the composition is partially purified by the following methods: a) degreasing whole eggs or yolk B) separating water-soluble fractions from defatted whole eggs or yolks; c) separating permeates below about 3,000 Daltons from the water-soluble fractions; and d) reducing the low The permeate fraction was separated at approximately 3,000 Daltons to recover the partially purified anti-inflammatory composition. 2. The anti-inflammatory composition according to item 1 of the application, wherein the chicken is hyperimmune with an immunogen. 3. The anti-inflammatory composition according to item 2 of the patent claim, wherein the immunogen vaccine includes a bacterial antigen. 〇: \ 4 «; \ 49495-4-93 () 409 DOQTSP This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)