JP3054194B2 - Immunosuppressive products - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Interaction with Related Patent Applications] This application is filed with US Patent Application No. 07/431639 (filing date 198).
(November 6, 2009) and US Patent Application No. 0
This is a continuation-in-part application of 7/177223 (filing date: April 4, 1988). U.S. patent application Ser.
This is a continuation-in-part application of No. 161039 (filed on February 26, 1988). U.S. Patent Application No. 07/161039 and U.S. Patent Application No. 0
No. 7/177223 is a divisional application of U.S. Patent Application No. 06/546162 (filing date: October 27, 1983)
It is a continuation-in-part application of 07/001848 (filing date January 9, 1987). US Patent Application No. 06/546162 is a continuation-in-part of US Patent Application No. 07/384625, filed June 3, 1982, now abandoned.

The contents of all of the above patent applications are entirely incorporated herein by way of explanation.

FIELD OF THE INVENTION This invention relates to the discovery of polypeptide fractions in milk containing peptides derived from allergens introduced into dairy animals. These fractions are useful for suppressing allergic reactions in humans and animals.

BACKGROUND OF THE INVENTION Allergic reactions in humans and animals have been studied extensively and the underlying immune mechanisms involved have been well understood. The common name for a molecule that causes an allergic reaction is allergen. There are many allergen species. Common examples include plant pollen, bee venom, house dust, animal desquamation,
And many food proteins. Many allergens are proteins or polypeptides in nature because proteins and polypeptides are generally more antigenic than carbohydrates or fats. Allergic reactions occur when IgE-type tissue-sensitizing immunoglobulins react with foreign allergens. When IgE antibodies bind mast cells and / or basophils and are stimulated by divalent antigens cross-linking antibody molecules, these specialized cells release chemical mediators of allergic reactions. Histamine, platelet activating factor, arachidonic acid metabolites, and serotonin are among the best known mediators of allergic reactions in humans.

The symptoms of an allergic reaction vary depending on where in the body IgE reacts with the antigen. If the reaction occurs along the respiratory epithelium, the symptoms are sneezing, coughing, asthma-like reactions. Abdominal pain and diarrhea are common when interactions occur in the gastrointestinal tract, as in the case of dietary allergies. For example, systemic reactions caused by bee stings are often serious and fatal.

A preferred, but often impossible, method for eliminating allergies is allergen avoidance. If unsuccessful, there are two types of medical means of controlling allergies, both of which have an effective rate of about 60-85% [Earth, K., Alagi, 37, 1-14 (1982)]. . The most common medical treatment for allergy is to treat the symptoms. Drugs known to block the effects of chemical mediators of allergic reactions (eg, antihistamines, etc.) are used to control the severity of allergic symptoms. However, these drugs do not help prevent allergic reactions and the release of chemical mediators, nor do they help prevent or eliminate allergic reactions upon subsequent allergen exposure.

Another means is to prevent allergic reaction by desensitization of the allergic host. This is accomplished by repeated small doses of the reactive allergen. Treatment is usually performed by subcutaneous injection of the allergen. It is believed that treatment with a reactive allergen, more appropriately called immunotherapy, increases the concentration of IgG-type antibodies to the allergen. IgG antibody is IgE
Compete with antibody allergen binding and neutralize and calm the tissue sensitizing IgE antibody response in some way, despite a clear correlation between antibody blockade and symptom improvement has not been clearly demonstrated, Or block [Mailing, HJ
Ed., Immunotherapy Position Paper, Allergy (Extra Number 6), Vol. 43, pp. 9-33 (1988)]. Although this theoretical interpretation is generally accepted, it has not been fully elucidated and does not fully reflect the complex interactions and individual phenomena associated with IgG reactions. Other effects of immunotherapy, including, for example, remission of symptoms, include suppression of IgE, increased blockade of IgA and IgG in secretions, decreased basophil reactivity / sensitivity, and decreased lymphocyte reactivity to allergens. Is mentioned. Not all of these changes can occur in any patient, and those changes associated with actual remission of symptoms have not been conclusively elucidated [Norman, PS, Journal of Allergy & Clinical Immunology, 75, 531-545 (19
85)).

Immunotherapy using reactive allergens is dangerous because the sensitized host is actually treated with molecules capable of eliciting an allergic response. Treatment is started with very low doses to avoid inducing a vigorous response.
The antigen concentration required to release 50% histamine from peripheral basophils in allergic patients varies 10,000-fold between patients [Norman, PS, Journal of Allergy.
And Clinical Immunology, 75, 531-545 (1985)]. If no side effects occur, a higher dose is administered. Injection can cause severe allergic reactions, so great care must be taken. If a severe reaction occurs, only an experienced physician can perform this procedure because immediate medical treatment must be given to control the symptoms of the allergic reaction. Since desensitization is an expensive, painful and time-consuming method [Earth, K., Allergy, Vol. 37, pp. 1-14 (1982)], and therefore is treated by this method only in the case of the most severe allergies [Mailing, HJ
Ed., Immunotherapy Position Paper, Allergy (Extra Number 6), Vol. 43, pp. 9-33 (1988)].

Modifications of allergens, ie, allergens that abolish the ability to induce a response by IgE, but retain the immunogenicity, ie, the ability to elicit a protective IgG response, have been studied for many years. In polymerized antigens, due to the latent antigenic determinant, the molecular concentration is low, thus reducing the chance of crosslinking on a weight basis,
They were evaluated based on the theory that diffusion into tissues would be more delayed and show lower antigenicity [Patterson,
R., Journal of Allergy and Clinical Immunology, 68, 85-90 (1981)]. Although normal IgM and IgG responses to antigens occur, allergen conjugates have been tested with various allergens in an attempt to selectively suppress the production of IgE antibodies [Lee, WY
Et al., Immunological Review, 41, 200-217 (19
78)]. Conjugates with glutamate / lysine copolymers can specifically act on IgE lymphocytes, suppressors and / or helper T cells, or a combination of these two mechanisms can occur [Riu, FT et al., Proc. Dings of the National Academy of Sciences of the USA, Vol. 76, pp. 1430-1434 (1979)].

Modifications to reduce the size of the allergen were also investigated to eliminate immunogenicity. It has been reported that limited proteolysis can indeed cause the appearance of suppressor determinants on allergen molecules [Mowatt, AM, Immunology, 56, 253-260 (1985).
Year)].

The use of enzymes that degrade the polypeptide fraction of the allergen and the parent protein is not new [King, T .;
P., Advances in Immunology, 23, 77 (1976)]. US Patent No. 4,469,677 reports the use of a polypeptide fraction prepared from an allergen to desensitize allergic humans and animals. A polypeptide fraction is prepared by digesting the allergen with a proteolytic enzyme. After enzymatic digestion, the enzyme and residual parent allergen must be removed from the polypeptide subfraction. The particular structure of the polypeptide fraction will depend on the enzyme used in the digestion process. However, this method is a synthetic operation that requires that an enzyme to be used for digestion be selected in advance. The polypeptide fraction is produced by controlled proteolytic digestion of a polypeptide allergen. Although this method has shown improvements over methods using reactive antigens, treatment still requires frequent painful injections.

The function of the polypeptide fraction that suppresses immune function is structure-dependent [Unanue, ER et al., Science, 236, 5
51-557 (1977), a review on the structural dependence of the polypeptide fraction of proteins on the immune response].

The structure of Michael's polypeptide fraction is limited because of the choice of enzyme based on the limited recognition that the enzyme is not always the most important.

Oral "tolerance" is a normal phenomenon,
It is a function necessary to respond to various foreign antigens ingested by the diet. Oral tolerance is initiated by a special class of T lymphocytes and their products,
Causes systemic suppression of IgE-dependent hypersensitivity reactions. Various responses of other components of the immune system have been reported to occur during induction of tolerance, and thus the true mechanism has not been clearly defined.

In animal studies, in response to intragastric administration of pollen extracts,
IgE production has indeed been found to be enhanced [Henderson, DC et al., International Archives of Allergy and Applied Immunology, 79
Vol. 66-71 (1986)]. Although this is a dose-proportional phenomenon, one study found that IgE increased between two studies of mice immunized orally or parenterally with 20 mg ovalbumin [Handson DG et al., International Archives. Of Allergy and Applied Immunology, 55, 256-532 (1977)]
And other studies that reduced it [Lafont, S. et al., Journal of Experimental Medicine, 155
Vol., Pp.1573-1578 (1982)].

One investigator has concluded that recognition of antigenic determinants differs between orally and parenterally induced suppressor T cells. Suppressor cells generated from dietary antigens can recognize different forms of antigen, but parenterally-derived suppressor T cells were specific for identifying the molecule to which they first responded [Moiwat, AM, Immunology,
56, 253-260 (1985)]. If this phenomenon were exhibited by the majority of antigens, it would further justify the use of oral immunotherapy.

A further problem with oral administration of allergens to allergic subjects is that a severe immune response can occur following treatment by this route. Anaphylaxis, recurrence, hives, rectal bleeding, and anaphylactic shock have been reported with oral immunotherapy [Platts Mills, TAE, Journal of Allergy.
And Clinical Immunology, 80, 129-132 (1987)]. Moreover, due to the effects of gastrointestinal enzymes on allergens, oral treatment requires significantly higher doses than parenteral treatment. However, since the individual response to ingested antigens can vary, more or less intact allergens can be provided to the gastrointestinal mucosa from the same dose administered to different subjects, increasing the risk of inappropriate dosage.

The non-allergenic fraction of the allergen can be administered orally without taking into account a severe immune response. However, for obvious reasons, oral administration of the polypeptide fraction of certain allergens has been described by Michael in US Pat. No. 4,469,677.
No.) and is not effective in causing desensitization. Studies of the immune response to other modified protein intakes are limited with a variety of different results. Collagen-induced arthritis in mice is a natural II
Inhibited by ingestion of type II collagen but not by ingestion of denatured molecules [Nagler Anderson, CL et al., Proceedings of the National Academy of Sciences of the US]
A, 83, 7443 (1986)]. In another study of autoimmune diseases, both pathogenic and non-pathogenic fragments and the decapeptide of myelin base protein were able to elicit immunosuppression in rats by oral treatment [Higgins, PJ
Et al., Journal of Immunology, Volume 140, 440-44
5 (1988)]. Studies in mice that evaluated the induction of oral tolerance in both native and modified ovalbumin showed that the suppression of delayed-type hypersensitivity to each form was the same [Mowatt, AM, Immunology, vol. 56, p. , Pp. 253-260 (1985)].

One factor that may be responsible for the inability of certain molecular fragments to effectively induce tolerance is the low pH of the stomach, which may further modify the polypeptide fraction to lose its ability to induce tolerance.

Another factor involved in the inability of an orally administered allergen fragment to induce desensitization to consequent exposure may be the intrinsic nature of the fragment itself.
In vitro synthesis of fragments requires specific enzymes and conditions that may or may not result in the preparation of a biologically optimal formulation. Fragments can be effective when suddenly exposed to the immune system by intravenous administration, but can be sufficiently susceptible to modifications that cause some changes in the gastrointestinal tract before being exposed to suitable immune system components Can destroy its effectiveness.

Thus, there is a need for oral formulations of allergen fragments that are sufficiently active to induce tolerance when administered orally, but are not allergenic. There is also a need for a method of producing such products.

SUMMARY OF THE INVENTION The present invention embodies the discovery of a product by a manufacturing method, a method of use, and processing. A common form of the product of the processing of the present invention is milk containing the polypeptide fraction of the allergen. A specific product of the processing of the present invention is a milk polypeptide fraction containing a polypeptide fragment of a specific allergen.

Specifically, the invention relates to milk produced by an immunized dairy animal, such as milk containing a non-immunoglobulin polypeptide fragment of an allergen used to immunize the animal.

The invention further relates to a food product containing the milk or an active fragment of the milk of the invention.

The invention further relates to oral vaccines administered to allergic subjects for the amelioration of allergic reactions in such subjects. Such a vaccine contains the milk of the invention, or an active fraction thereof.

The invention further relates to a method of producing an immunosuppressed product comprising immunizing a lactating animal and, after the animal has reached an immune state, collecting milk from such animal.

The present invention further relates to a method for desensitizing a subject to an allergen, comprising administering to the subject the milk of the present invention, or an active fraction thereof, in an amount and for a time sufficient to produce an immunosuppressive effect. Things.

The present invention further comprises an active polypeptide fragment of the present invention in a subject in need of desensitization to an allergen, in an amount effective to treat and suppress the allergic response and allergic response of such subject. And a pharmaceutical composition that provides the desired fraction.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention provides an oral formulation for desensitization, ie, an oral vaccine that is significantly medically beneficial to the subject taking it. By subject is meant a human or other animal in need of allergic desensitization by the method of the present invention. The oral vaccine of this invention can be self-administered, has less pain than vaccines that must be injected,
There is no cost. Subjects who have received the oral vaccine of this invention are exposed to a processed and allergen that has been derivatized in such a way that such subjects can acquire tolerance to the allergen.

The milk of the present invention, or an active fraction thereof, can be administered in any form, either a powder or a liquid, which retains the allergic response properties of milk. Any food product incorporating such an active fraction of milk, for example, skim milk,
It can be provided as yogurt, and cheese.

The "active fraction" of the milk of the present invention refers to the allergen extracted from the milk of the present invention and used to immunize an animal that provides such milk, present in a derivatized and processed form. Accordingly, it represents a formulation or composition possessing the beneficial allergic reaction-suppressing properties of the unextracted milk of the present invention.

The derivatized, processed form of the allergen of the invention is present in the milk of dairy animals immunized against the allergen. The advantage of administering the polypeptide fraction of the present invention to the subject instead of administering the unprocessed allergen itself is different from the unprocessed allergen, in that the allergic reaction-inhibiting polypeptide fraction of the milk of the present invention is different from the unprocessed allergen. Non-allergenic. Desensitization of the subject who has consumed the milk of the present invention occurs, but the immune system in the milk-producing host processes the allergen into a non-allergenic form, which results in severe allergic reactions in such subjects. There is no danger.

The allergen used as a source of an immunosuppressive active polypeptide can be any allergen that can develop an immune system response in a mammal that processes such an allergen. If the allergen cannot elicit an immune system response in one mammalian species, another species may be used.
In a preferred embodiment, the dairy cow, a cow that produces milk, is used to treat the allergen. However, any mammalian species can be used, including humans and animals such as sheep, pigs, and horses.

The allergen preparation may contain a mixture of allergens, or only one specific allergen. Examples of allergen mixtures that can be protected by administration of the milk preparations of the present invention include pollen of various trees and grasses, venoms from animals and insects, and mixtures of food allergens, and the like. Examples of specific allergens that can be protected by administration of the formulations of the invention include, for example, ragweed, bee venom,
Examples of food allergies include wheat protein.

The method for producing a polypeptide fraction of the present invention, when used to treat a host to be allergic to such an allergen, is unique to the parent allergen, such as a subfraction that exhibits immunosuppressive activity. Produce subfractions. "Immunosuppressive activity" refers to the ability of a composition to reduce allergic symptoms or reactions in an allergic subject to whom such a composition has been administered. "Allergic symptoms" means that an individual responds to exposure to a substance that is allergic to the individual and is induced by the individual, for example, allergic hyperemia, headache, hyperventilation, itching, swelling, sneezing, wheezing. Physiological responses or conditions such as coughing, rhinorrhea, and decreased olfaction and taste.

The milk composition of the present invention, and the polypeptide fraction produced therefrom by the method of the present invention, lack the allergenicity of the parent allergen (the allergen used to immunize the lactating animal). Has the ability to desensitize a host to which such a composition has been administered.

It is not necessary that the dairy animal be induced or maintained in the milk-producing hyperimmune state of the invention. Furthermore, since the active composition of the present invention does not provide protective inhibitory activity by means of antibodies,
It is not necessary to induce the milk-producing state of the present invention to an antibody-producing state. It is only necessary that the animal receiving the allergen provide sufficient time to process such allergen into a form that secretes it into the milk of the animal.

Since immune cell processing of an allergen is one of the most written stages of the immune response, milk containing the active polypeptide of the invention can be obtained relatively early after administration of the allergen, for example, 24 hours after administration of the allergen. ~ 1 week. Dairy cows that have been administered an allergen according to the methods reported herein will provide such active polypeptide in their milk 24 hours after administration of the allergen.

Any amount of allergen that can result in the appearance of an active polypeptide of the invention in the milk of the animal can be administered to the dairy animal. If the amount of active polypeptide in the milk of the animal to which the allergen is administered is too low to provide an effective effect to the subject in need of desensitization, the active polypeptide should be concentrated in a more concentrated form. Can be provided. Such concentration can be performed using techniques known in the art, for example, by salt precipitation, evaporation, or lyophilization of the composition of the invention.

Active polypeptide-containing formulations are combined from one or more allergen-treated animals to provide a subject in need of desensitization with a composition containing various polypeptide subfractions from different genetic predispositions, Combinations can be made from more than one allergen-administered animal species.

It is not necessary to induce a dairy animal to an antibody-producing state or a hyperimmune state, but if the milk of the present invention and the continuous production of milk containing the active polypeptide of the present invention are not detrimental to the milk, the milk is This condition can be achieved by repeatedly administering the allergen to the producing host.

Although the processing mechanisms of the immune system have not yet been fully elucidated, there is no intention to support this theory, but exogenous proteins or polypeptides are ingested and eaten poorly by specialized cells of the immune system Most notable are macrophages and neutrophils. After administration to a lactating animal, enzymes within the immune cells destroy the protein or polypeptide allergen and are used interchangeably herein in the "processed" or "subfractionated" form of the allergen. Derivatization.

The subfractions produced by immune cells are unique and cannot be produced outside of the natural host. Its uniqueness is based firstly on the structure of the parent allergen and secondly on the combination of enzymes in the allergen-treated cells of the mammalian host. Combinations of enzymes expressed in mammalian allergen-treated cells are determined genetically. Other species have unique combinations of enzymes. For example, the peptide fraction of allergen treated by immune cells of bovine species is similar, but not identical, to the polypeptide fraction treated by immune cells of other species. It is this genetic specificity that makes the polypeptide produced by the method of this invention unique and different from polypeptides produced by any other method.

Special immune cells, called helper cells, help macrophages present processed antigens to T and B cells. The course of the allergen subfraction entering the milk through the mammary gland by and / or from the macrophage and through the blood may further modify the structure of the allergen polypeptide present in the milk. This natural biological method of processing environmental antigens and transporting natural immunosuppressive factors into milk provides a mechanism for dairy cows to deliver immunoprotective factors to calves by breast milk. These milk immunity factors protect lactating pups of such animals from allergic reactions. Nature has evolved to find a way for mothers of species to determine the potential allergen environment. The immune system then turns environmental allergens into factors that protect against allergic reactions. The dam sends the immunoprotective factor through lactation to the nursing infant. The main focus of the discovery of the present invention is that humans have access to this natural biological process that turns a known allergen into an immunoprotective polypeptide fraction that is useful in the prevention and treatment of allergic reactions. is there.

The immunosuppressive factor in milk is the polypeptide sub-fraction found in milk, not the antibody found in milk. Although antibodies may be useful for treating allergies, milk antibodies are not a product of this invention.

The milk of animals treated according to the method of the present invention contains a specific and unique polypeptide subfraction of the allergen used to treat the host. This milk lacks the intact, ie, non-derivatized, parent allergen. The uniqueness of these polypeptide fractions derives from the specific enzymatic breakdown by specialized immune cells of the host. Blood and breast tissue enzymes may also play a role in modifying the chemistry of the allergen prior to entering milk. Milk containing these unique non-reactive polypeptides is useful as an oral immunization that, when ingested by other species, including humans, induces protection and / or tolerance. When isolated from milk, the milk allergen polypeptide fraction is useful for suppressing an allergic reaction to a representative parent allergen.

Allergens can be administered to dairy animals by various routes of administration including oral, rectal, vaginal, intramuscular injection, subcutaneous injection, intradermal injection, transdermal administration and the like. Although any route of treatment can be used for allergen administration, the preferred route of treatment is by intramuscular injection of the allergen into the host species. When an allergen enters the body of a dairy animal, it is processed by the immune system of the milk-producing host.

When an allergic individual or animal is treated with a composition of the invention, protection against the parent allergen is established in the individual or animal without the risk of side effects that can occur when using the parent allergen molecule. Examples of protein or polypeptide allergens that can be processed by mammalian lactation organs include weeds, grasses, tree pollen allergens, animal venoms and toxicants, and various food allergens. In a preferred embodiment, bovine lactating organs are used. In theory, any allergen can be processed by mammalian, especially bovine, macrophage systems to produce immunosuppressive polypeptide subfractions.

The method of the present invention is advantageous because it uses the full spectrum of processing enzymes available in allergen-treated cells of the mammalian immune system. Second method of the present invention
The advantage of is that the complete spectrum of the polypeptide fraction is contained in the immunosuppressed milk product, and when contained in a natural milk carrier, the immunosuppressive allergen fraction is orally active. In the method of the present invention, there is no need to select a specific enzyme for decomposing the allergen in a test tube or to control the enzymatic digestion treatment. There is no need to separate from Complete antigen processing occurs naturally in lactating animals.
Mammalian genes select enzymes and the natural metabolism of mammals keeps reaction conditions at optimal levels of antigen processing.

By any technique by which a peptide can retain its biological activity, i.e., by which such a peptide can possess the ability to suppress an allergic reaction, or by a combination of such techniques,
The peptide fraction of the invention can be monitored, purified and isolated. Such techniques are known in the art and include adsorption chromatography, gel chromatography, especially dextran, polyacrylamide, or agarose gel.
Gel chromatography using matrices, ion exchange chromatography, such as "SP-Sephadex" (containing sulfopropyl functional groups derivatized to dextran), "AG50" (styrene divinyl
Containing sulfonic acid derivatized to benzene),
And strong cation exchange resins such as "Bio-Rex 40" (containing sulfonic acid groups derivatized to a phenolic matrix), for example "CM-Sepharose"
Weak cation exchange resins such as (containing a carboxymethyl functional group derivatized to dextran), "Bio-Rex 70" (containing a carboxyl functional group derivatized to a phenolic matrix), for example "QAE-Sephadex" (containing diethyl- (2-hydroxypropyl) -aminoethyl functional group derivatized to dextran), "AG1" (tetramethylammonium ion derivatized to styrene-divinyl-benzene) Exchange resins such as "DEAE-Sephadex" (containing a diethylaminoethyl functional group derivatized to dextran), "AG-3" (an epoxyamine matrix). Containing a tertiary amino function derivatized to
Weak anion exchange resins such as "CM-cellulose" (containing a carboxymethyl functional group derivatized to cellulose), "P-cell" (containing a phospho functional group derivatized to cellulose). Cation exchange resin such as "DEAE-cellulose"
(Containing diethylaminoethyl functional groups derivatized to cellulose), "PEI-cellulose" (containing polyethyleneimine functional groups derivatized to cellulose), "DEAE (BND) -cellulose" (to cellulose) Intermediate strength anionic exchanges such as derivatized benzoylated-naphthoylated diethylaminoethyl functions), "PAB cellulose" (containing p-aminobenzyl functions derivatized to cellulose) Utilizes a resin and an exchange resin that provides a mixture of functional groups such as, for example, "AG501" (containing two functional groups of sulfonic acid and tetramethylammonium ion derivatized to a styrene-divinylbenzene matrix). Chromatography techniques, including ion exchange chromatography, etc. It is, but not be limited only to this. High performance liquid chromatography can also be used to separate the peptides of the invention.

To enhance or provide additional properties to the active polypeptide-containing compositions of the present invention, particularly those that require immunosuppressive treatment, to enhance the ability of the active polypeptide to promote the amelioration of the effects of the allergen, Other compounds can be chemically or genetically linked to such active polypeptides.

Dosages and dosage regimens for the compositions of this invention can be readily determined by one of ordinary skill in the art. In general, administration of the active peptide-containing composition depends on the type of allergen used, the age of the subject to be treated, the condition of the subject to be treated, the type of allergy to be treated, the type of co-treatment, if any, Taking into account the physiological tolerability of the composition, the number of treatments and the nature of the desired effect, sex, duration of symptoms, contraindications, if any, and other conditions that can be adjusted as desired by the individual's physician. change. Administration can be carried out in one or more applications until the desired result is obtained.

The compositions of the present invention can be administered in addition to any suitable pharmacological carrier for administration. They can be administered in any form that can prevent, alleviate, prevent, or cure the allergic reaction in humans and animals.

The preparation for parenteral administration of the active polypeptide-containing composition of the present invention includes sterile water or non-aqueous solvent, suspension, emulsion and the like. Illustrative non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils, fish oil, and organic esters for injection. Aqueous carriers include water, water-alcohol solutions, suspensions and emulsions containing saline, sodium chloride solution, Ringer's dextrose solution, sodium chloride solution with dextrose, lactose-containing Ringer's solution, or fixed oils. And a buffered pharmaceutical parenteral carrier. Intravenous carriers include liquids,
Nutritional replacement fluids and electrolyte replacement solutions based on Ringer's dextrose and the like.

Also, the compositions of this invention can be administered by pump or in sustained release form.

Administration in a sustained release form is more convenient for patients when repeated dosing is indicated for a long period of time.

The active polypeptide-containing composition of the present invention can be used in dosage forms such as tablets, capsules, powder packets, or liquid solutions for oral administration.

The pharmaceutical composition of the present invention is produced in a manner known per se, for example, by conventional mixing, granulating, dragee-making, dissolving, lyophilizing or similar operations.

As described above, the present invention has been generally reported. Hereinafter, the materials and methods used for practicing the present invention will be described in detail with reference to examples. These examples are intended to illustrate the invention and do not limit the invention in any way.

EXAMPLES Example 1 Immunization of Dairy Cows and Collection of Immune Milk The immunogen for preparing milk products for allergic symptoms reduction was a commercial allergen purchased as a 1: 100 (weight: volume) preparation. The extracts contain the mixture pooled in the proportions in the table below.

Dilute the pooled mixture with an equal volume of sterile saline. During normal feeding periods, dairy cows are injected intramuscularly with 5 ml of the formulation at two week intervals, and the milk is collected and powdered. Milk collection after treatment
Start from 24 hours.

Example 2 Fractionation of Milk Fraction Containing Processed Allergen 20 liters of fresh milk from hyperimmunized dairy cows was run on a cream separator (DeLaval 102) to separate fat.

The resulting 16 liter skim milk was ultrafiltered using a hollow fiber diafiltration / concentrator to obtain high molecular weight species (1000
(00 daltons or more). The concentrator has two 1000
Equip a 00 dalton molecular weight cut-off cartridge. The skim milk was processed at a pump speed of 80, inlet and outlet pressures of 30 and 25 psi, respectively.

12 liters of filtrate (<100,000 Daltons) that flowed out of the cartridge at a flow rate of 4 liters per hour were stored frozen or lyophilized.

Using this same method, milk from non-hyperimmunized dairy cows can be fractionated.

Example 3 Desensitization of Allergic Subjects Approximately カ ッ プ cup of immunized milk powder was dissolved in 8 ounces of water, reconstituted, and taken by human volunteers. The 200 volunteers were 4-87 years old and took milk products for 1-172 months. Of the 168 allergic subjects who responded indicating disease status, 138 (82.1%) showed improvement while taking milk products. Ninety-four percent of those who reported efficacy said improvements had occurred within one month of starting taking the dairy product. Specific effects reported were reduced hyperemia, reduced or eliminated headache, ease of breathing, itching, swelling, sneezing, wheezing, coughing, rhinorrhea, improving olfaction and taste. Many subjects reported a recurrence of symptoms following withdrawal of milk products, and some subjects discontinued allergic immunotherapy after taking milk products.

Having completely described the invention, those skilled in the art will recognize that the scope of the claims
Or, it will be appreciated that it may be practiced within a wide range of corresponding conditions, parameters, etc., without affecting any of its embodiments.

Continued on the front page (58) Fields surveyed (Int. Cl. ⁷ , DB name) A61K 39/35 A61P 37/08 A23C 9/00 CAPLUS (STN)

Claims (8)

(57) [Claims] Claims: 1. A subject having a non-antibody fraction of milk, wherein the non-antibody fraction of milk is allergic to an allergen. Allergic symptoms are ameliorated, and the above fraction is (a) administering an allergen to a lactating animal, (b) collecting milk from the animal in (a) part, and (c) eliminating a molecule larger than 100,000 daltons The milk from part (b) is filtered through a filter, and the filtrate from the filtration from part (d) and part (c) is collected, wherein the filtrate is produced by an operation that includes the above-mentioned active fraction. Composition. 2. A food product containing the composition according to claim 1. (3) administering the allergen to a lactating animal; (b) collecting milk from the animal of (a); and (c) filtering the milk of (b) with a filter which excludes molecules larger than 100,000 daltons. Filtering the milk, collecting the filtrate from the filtration of part (d) (c), wherein the filtrate contains a non-antibody fraction of milk, comprising a method comprising containing the active fraction described above. ,
A method for producing a pharmaceutical composition, wherein the non-antibody fraction of the milk is intended to improve the allergic symptoms of the subject to the allergen when the subject ingests the fraction in a subject having an allergy to the allergen. . 4. The method according to claim 3, wherein the dairy animal is a cow. 5. A method of desensitizing a subject to an allergen comprising orally administering a food product to the subject in an amount and for a time sufficient to effect allergic symptoms of the subject against the allergen, A desensitization method comprising the food product containing a non-antibody fraction of milk of a dairy animal immunized with the allergen. 6. The food product is milk, skim milk, yogurt,
6. The method of claim 5, wherein the method is selected from the group consisting of: and cheese. 7. The method according to claim 5, wherein the food product is in powder form. 8. The method according to claim 5, wherein the food product is in a liquid form.