KR20010043426A - Anti-Diarrheal and Method for Using the Same - Google Patents

Abstract

The present invention relates to food and methods for treating and preventing diarrhea in a subject animal suffering from or susceptible to diarrhea. This method includes administering an egg product obtained from a hyperimmunized bird to a subject animal.

Description

Anti-Diarheal and Method for Using the Same}

Diarrhea is a worldwide problem for individuals of all ages. Diarrhea is a common symptom that can disrupt life and threaten life to a minimum. Acute diarrhea can be caused by various germs, functional disruption of intestinal function, and drug-related side effects.

Some symptoms of diarrhea are infant diarrhea (eg rotavirus-induced), infant diarrhea, microbial induced diarrhea (ie, food poisoning), most gastrointestinal disorders, and diseases that indirectly affect the gastrointestinal tract. Infants and toddlers with diarrhea have major problems because they can become severely dehydrated and require fluids and medications.

Diarrhea is also common in cancer patients and may interfere with cancer treatment [Ippoliti, 1998]. Chemotherapy, radiation therapy, surgery, graft-versus-host disease (GVHD), bone marrow transplantation, or infection can also induce diarrhea. In addition, bacterial pathogens can also cause diarrhea syndrome with the effect of a range of severe tissue damage or a perceived extent of damage deficiency. Enterotoxin-producing E. coli, which causes acute and severe diarrhea, has this effect by producing potential toxins that act by modulating biological activity in epithelial cells [Isaacson, 1998].

The current treatment of diarrhea consists of pharmacological involvement of the antibiotic or pathophysiological action of the causative microorganism. Antidiarrheal agents reduce diarrhea symptoms (diffuse density of bowel movements, frequency of bowel movements and excessive bowel movements) by affecting intestinal passage, mucosal transport or lumen content [Schiller, 1995]. Opiate agents, such as loperamide, are the most useful antidiarrheal agents and work in combination with intestinal transit inhibition, pre-absorption and antisecretory effects. Other useful pharmacological therapies include the use of alpha-adrenergic agents such as clonidine and somatostatin analogs. These drugs can modify mucosal transport in addition to slowing passage, but their clinical use has been limited because of their nonspecific action. Adsorbents (Kaopectate), bismuth (Pepto-Bismol) and stool modifiers are often used as counter drugs, but most of their efficacy other than pepto-bismol has not been demonstrated.

Safe and effective natural antidiarrheal agents will be very useful for acute and chronic diarrhea symptoms.

<Summary of invention>

The present invention is based on the inventors' finding that the egg or egg product, and particularly the egg product of the over-immunized algae, has a branch activity when administered to a subject animal to prevent or reduce diarrhea in the subject animal.

In particular, the present invention relates to a method for treating and preventing diarrhea in a subject animal suffering from or susceptible to diarrhea, comprising administering to the subject animal an effective amount of an egg product.

The invention also relates to a composition comprising beneficial properties for preventing or reducing diarrhea in a subject animal suffering from or susceptible to diarrhea, comprising a product obtained from an animal that is overimmunized with one or more immunogens. .

The present invention relates to products and methods for treating and preventing diarrhea. More specifically, the present invention relates to natural foods that, when administered to a subject animal, treat and prevent diarrhea in the subject animal.

The hyperimmune egg product of the present invention is useful for the treatment and prevention of diarrhea in a subject animal when administered to the subject animal.

Terminology:

The term "diarrhea" means abnormal frequency and liquidity of bowel movement resulting from an imbalance between intestinal absorption and secretion.

The term "hyperimmune" means exposure to one or more immunogens, such as an immune response that is elevated and maintained above a natural, unexposed state.

The term "al" or "alk product" means each egg (table, over-immunized or otherwise) or any product or fraction derived therefrom, respectively.

The term "table egg" or "table egg" means each egg obtained from an egg-producing animal, or any product or fraction derived therefrom, that is not maintained in a hyperimmune state, respectively.

The term "hyperimmune egg" or "hyperimmune egg product" means all eggs or any product or fraction derived from an egg producing animal, each maintained in an overimmunized state.

The term “supernormal level” means a level above that found in eggs of egg-producing animals that have not remained hyperimmune.

The term “immune antigen” refers to a substance capable of eliciting humoral antibodies and / or cell-mediated immune responses in addition to immunological resistance. The term refers not only to the ability to stimulate immune response but also to its reaction with a product, for example an antibody.

The term “combination induced immunoantigen” refers to a novel method of generating molecular diversity among immunogens by combination synthesis.

The term "bioengineering immunoantigen" refers to an immunoantigen obtained through gene cloning techniques and genetic rearrangement methods that allow the insertion of nucleotide encoding genes capable of generating molecules with immunoantigen properties.

The term "genetic vaccine" refers to a nucleic acid vaccine that is generally produced by recombinant technology and capable of eliciting an immune response.

The term "treatment" means the onset of a symptom (including pain) of a disease and / or a pathological origin of the disease is delayed or completely prevented, or if present, an improvement or complete elimination of the syndrome. For example, hyperimmune egg products not only relieve symptoms of diseases of humans and other mammals, but also act as prophylactic agents that interfere with the presence of the disease in recipients to treat arthritis and / or autoimmune diseases.

The term "prevention" means that the progress of the disease is reduced and / or eliminated, or the onset of the disease is eliminated.

The term "administration" means any method orally, intranasally, parenterally (intravenously, intramuscularly or subcutaneously), rectally or topically to provide a substance to a subject.

The term "animal" means the animal kingdom definition.

The term "target animal" means an animal that functions as an egg or egg product producing animal.

The term "subject animal" means the animal to which the egg or egg product produced by the target animal is administered.

The present invention

The hyperimmune egg product of the present invention is useful for treating and preventing diarrhea in a subject animal when administered to the subject animal.

The products and methods of the present invention are particularly concerned with the use of natural foods, hyperimmune egg products, for the treatment and prevention of diarrhea and associated diseases in a subject animal. Being natural, this food can be used for the treatment and prevention of diarrhea and associated diseases without side effects. Animals allergic or hypersensitive to eggs may not be able to digest the hyperimmune egg product of a particular dosage form.

In a preferred embodiment, the present invention comprises an hyperimmune egg or egg product obtained from an egg-producing animal, more preferably an alga, hyperimmunized with one or more immunogens. The hyperimmune egg product is preferably administered orally to the subject animal even if the hyperimmune egg or egg product can be separated into more potent fractions and then administered to the subject animal in various forms.

Hyperimmune eggs or egg products of the present invention are particularly useful when administered to a subject animal, including infant diarrhea (eg, including viral or bacterial induced diarrhea), infant diarrhea, adult organ induced, and gastrointestinal disease induced diarrhea, etc. It is effective in treating and preventing all forms of diarrhea and associated diseases, including but not limited to such acute and chronic diarrheal diseases.

Hyperimmune egg product

Hyperimmune egg products can be produced from any egg-producing animal. Preferred are bird species, ie animals that are birds. Home poultry is preferred for the new species, and other varieties such as turkeys, ducks, and geese are also suitable sources of hyperimmune eggs.

When such an egg-producing animal is subjected to an immune specific state, for example, by periodic full explosive administration of an immunogen, the animal has eggs that have useful properties to treat and prevent diarrhea in the subject when consumed by the subject. Will generate

Such hyperimmune specific conditions are preferably achieved by administering a sufficiently high amount of a periodic protonic drug following initial immune administration followed by a specific immunogen or mixture of immunoantigens. The preferred dosage of the full explosive should be at least 50% of the dosage required to produce the bird's primary immunity. Thus, even when the algae are in a state commonly referred to as the immune state, they are below the initiation predose dosages that do not produce characteristics in the eggs of the algae.

Knowing the requirements for the development and maintenance of hyperimmune conditions, those skilled in the art can vary the immunogen dosage depending on the egg-producing animal and species used to maintain the animal in a hyperimmune condition.

Hyperimmune conditions are preferably generated with any immunoantigen or combination of immunoantigens. Hyperimmunity is preferably achieved by multiple exposures to multiple immunogens, multiple exposures to a single immunogen, or a single exposure of a library of immune antigens.

In addition to immunization with naturally occurring immune antigens, immunization can also be performed using immunogens synthesized by combinatorial chemistry. The basic method is to associate multiple combinations of chemical structural blocks that produce a diverse molecular population. Several methods have been described in oligomeric libraries [Fodor, S. et al., Science 251: 767 (1991); Houghton, R. et al., Nature 354: 82 (1991)], as well as solid and solution phase combination synthesis as well as small organic molecules [Bunin, B. & Ellman, J., J. Am. Chem. Soc. 114: 10997 (1992). Rapid multiple peptide and oligomer synthesis can function as a combination induced immunoantigen. Alternatively, organic structural blocks can be added to the improved immunogenic backbone molecules in a combinatorial manner.

Other types of hyperimmune egg producing animals may be used in place of immunoantigen vaccines, including the use of genetic vaccines. In particular, any DNA constructs (generally composed of promoter regions and immunoantigen coding sequences) will attract immune responses. Genetic vaccines include immunoantigen-coding vectors, fragments of naked DNA, plasmid DNA, DNA-RNA immunoantigens, DNA-protein conjugates, DNA-liposomal conjugates, DNA expression libraries, and viruses and bacteria delivered to generate immune responses. It consists of DNA. DNA delivery methods include, in particular, particulate bombardment, direct injection, viral vectors, liposomes and jet injection. When applying this method of delivery, smaller amounts may be needed, and in general a longer incubation period will result. Using this genetic method, the preferred method of injecting DNA into algae is to intramuscularly inject DNA into the pectoral muscles.

DNA delivery methods include, but are not limited to, particulate bombardment, direct injection, liposomes, jet injection [Fynan, E.F. et al., Proc. Natl. Acad. Sci. USA 90: 11478-11482 (1993). Known or unknown immunogens, promoter regions (particularly CMV cauliflower mosaic virus) and nucleic acids encoding SV40 bacterial origin can be replicated in bacteria to produce plasmid DNA used for DNA injection. Although parenteral administration of DNA via multiple routes is effective for chickens, the preferred method is intramuscular injection into the pectoral muscles. Vaccine testing was performed on eggs, preferably chickens, which lay eggs. Repeated immunizations were performed at 1 to 2 week intervals for 6 months.

The amount of DNA used is generally preferably 50 to 300 μg of DNA in saline for direct administration. The particle impact, 2.5 M by the addition of CaCl ₂ is preferably 4 to 100 ㎍ DNA was co-precipitated onto gold beads. Repeated immunoassays can be performed intradermal by accelerating DNA coated particles into living animals.

Hyperimmune course

The steps described below are examples of preferred processes used to raise an egg-producing animal into an advanced immune state in which the obtained hyperimmune egg or egg product can be administered to a subject.

1. Select one or more immunogens.

2. The first immunization method elicits the immune response of egg-producing animals.

3. Induce and maintain hyperimmune status by administering an appropriate dose of the immunoantigen full explosives vaccine.

Step 1: Any immunoantigen or combination of immunoantigens can be used as a vaccine. The immunoantigen can be bacterial, viral, protozoan, fungal, cellular, or any other substance to which the immune system of an egg-producing animal responds. It is important at this stage that the immunogen (s) should be able to induce immune and hyperimmune conditions in egg-producing animals. Although only one immunoantigen can function as a vaccine for the method of the present invention, any preferred vaccine includes enteric bacilli and bacteroides, pneumococci, Pseudomonas ( Pseudomonas, Salmonella, Streptococci, Bacillus, Staphylococci, Neisseria, Clostridia, Mycobacteria, Actinomyces Chlamydia Chlamydia And a mixture of multivalent bacterial and fungal immunogens selected from the family of immunogens such as Mycoplasma. Viral immunoantigens are preferably selected from immunogens such as Adenovirus, picornavirus and Hepes virus, and will also work with other immunogens.

In a separate embodiment, a multivalent vaccine called Series 100 (S-100) is used. The bacteria included in the S-100 vaccine are as described in Table 1 of Example 1. Vaccines are already described in US Pat. Nos. 5,106,618 and 5,215,746 assigned to Stolle Research and Development Corporation.

Step 2: The vaccine can be a dead or live supervised vaccine and can be administered by any method that elicits an immune response. The immunoassay is preferably performed by intramuscular injection of an immunogen. Intramuscular injection into algae is preferably injected into the pectoral muscles. The dosage is preferably 0.05 to 5 mg of the immunoantigen vaccine. Other methods of administration that can be used include intravenous infusion, intraperitoneal infusion, intradermal, rectal suppository, aerosol or oral administration. If DNA technology is used in the hyperimmune process, a smaller amount, typically 300 μg, is required.

Numerous methods known to those in the field of immunology can determine whether the vaccine induces an immune response in egg-producing animals. These examples include enzyme-linked immunosorbent assays (ELISA), tests for the presence of immunoantigen-stimulating antibodies, and tests designated to assess the ability of immune cells to respond to immune antigens in a host. The minimum dose of immunogen required to induce an immune response depends on the type of adjuvant used and the type of preparation of the immunogen (s) as well as the type of vaccine used, as well as the type of egg-producing animal used as a host. .

Step 3: The hyperimmune condition is preferably induced and maintained in the target animal by repeating the appropriate dose of full explosive at fixed time intervals. The time interval is preferably a 2-8 week interval for 6-12 months. However, it is essential not to administer the full explosives to induce immune resistance. This process is well known in the art.

Other hyperimmune maintenance procedures or a combination of procedures such as, for example, intramuscular injection for primary immunization and intravenous injection for pre-exposure injection may be used. Additional procedures include simultaneous administration of microencapsulated liquid immunogens, or intramuscular infusion for primary immunization, and full explosive administration by oral or parenteral administration by microencapsulation means. Several combinations of primary and hyperimmune are known to those skilled in the art.

<Preparation and Administration>

Once the egg-producing animal is sufficiently overimmunized, it is desirable to collect and process eggs from such animals to produce the hyperimmune egg product in an administrable form. The hyperimmune egg product can then be administered to the subject.

The subject animal is administered the egg and / or egg product of the invention by any method of treating or preventing diarrhea in the subject animal. It is preferred to administer directly to the egg or any derivative of the egg. Eggs and egg yolks are natural food ingredients, non-toxic and safe.

In one embodiment, the egg product of the present invention is incorporated into a food supplement. Preferred methods of making the eggs of the present invention incorporated into food supplements include drying the eggs into egg powder. Various methods of drying eggs are known, but spray drying is the preferred method. Egg spray drying methods are well known in the art.

Dried egg powder may be incorporated into beverages in various administrable forms, such as in the form of beverages, such as protein powders, vitality beverages, protein supplements and any other nutritional, athlete-related products. Egg powder may also be used in bake mixes, power bars, candy, cookies and the like. Alternatively, the egg powder may be administered in a capsule form as it is. Other examples of egg preparation include making omelets, ripening or ripening eggs, baking eggs, and if necessary, raw or prepared as liquid eggs.

Finally, it is generally known in the art that egg yolk and / or egg white include drug (s) resulting in the beneficial properties observed and described above. Further separation provides more effective fractions, or allows undesirable components to be removed, and in other ways, for example parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, intranasal, oral or It will be clearly appreciated by those skilled in the art that it allows for topical administration. This further separation will allow the encapsulation product and pharmaceutical composition with the eggs or fractions thereof to be prepared.

To determine the efficacy of the hyperimmunized egg product for treating and preventing diarrhea, we used an animal diarrhea model. The data obtained suggest that a high dose of hyperimmunized egg product can prevent and treat diarrhea in a well known animal model, referred to as the administration of castor oil described below (see Example 2).

Inhibition of castor oil-induced diarrhea has been used in the development of antidiarrheal agents such as diphenoxylate and loperamide in animal models and has been known as a good predictor of human adipose activity for more than 25 years (Awouters et al., 1974). This study shows that acute administration of eggs from chickens immunized against various immunogens can inhibit castor oil induced diarrhea.

The mechanism of the action of diarrhea in castor oil is complex. Castor oil must first be metabolized to ricinolic acid in the lumen of the intestinal tract. Ricinolic acid subsequently causes a significant increase in total fluid secretion and intestinal electrolytes resulting in diarrhea. Castor oil action mechanism was recently studied by Mascolo N et al., Who discovered the activation of platelet activating factor (PAF) in duodenal tissue by secreted nitric oxide (NO) in response to castor oil.

This study detailed in Example 2 below shows that long-term pretreatment is not required to show the activity of the hyperimmune egg product and is effective when orally administering the hyperimmune egg product of the monoclonal dosage form. Therefore, the conclusion of the present inventors is that the hyperimmune egg product is a safe and effective therapy for alleviating acute or chronic diarrhea regardless of the cause and can be usefully added to the therapy of diarrhea conditions. This study shows that when administered hyperimmune egg product in oral monoclonal dosage form for two days, it will significantly inhibit castor oil-induced diarrhea in mice.

In the treatment and prevention of diarrhea disorders, the hyperimmune egg product is preferably administered to a subject in an amount that is immunologically effective for the treatment and prevention of certain disorders. In castor oil studies, there was a dose-related effect at doses of 1, 2 and 4 g / kg. However, the dosage and duration of administration will depend on the particular condition of whether the condition has developed in the subject. The hyperimmune egg product is preferably provided in any amount necessary and effective to treat and / or prevent the condition and its symptoms. For example, in some cases, a daily dose of hyperimmune eggs (or up to several equivalents of the entire hyperimmune egg product) may be administered to a subject depending on the particular condition of the condition. Can be. More effective fractions from hundreds of eggs can be isolated and concentrated by methods well known in the art.

It is very important to point out that the egg products of the present invention are safe and nontoxic, ideal for long term use and have no side effects except for those who are allergic to eggs. Egg products may be administered orally, either alone or in combination with drug therapy, for long term use for diarrhea-related disorders.

Advantageous properties of the invention can be observed with reference to the following examples illustrating the invention.

<Example 1>

Preparation of the S-100 Vaccine

The bacterial cultures containing the broad range of bacteria listed in Table 1 obtained from the American Type Culture Collection were reconstituted with 15 ml of medium and incubated overnight at 37 ° C. Once well grown, about half of the bacterial suspension was used to inoculate 1 liter of broth incubated at 37 ° C.

After good growth was seen in the culture, the suspension was centrifuged for 20 minutes to remove the medium to obtain bacterial cells. The obtained bacterial pellet was resuspended in sterile saline solution and the bacterial samples were centrifuged three times to wash the medium from the cells. After the third sterile saline wash, the bacterial pellet was resuspended in a small amount of two distilled water.

The medium-free bacterial suspension was killed by placing it overnight in a glass flask in an 80 ° C. water bath. When broth cultures were tested with small amounts of dead bacteria, they were incubated at 37 ° C. for 5 days to verify that the bacteria were killed and checked daily for growth.

Dead bacteria were lyophilized until dry. Dry bacteria were then mixed with sterile saline solution to a concentration of 2.2 × 10 ⁸ bacterial cells / ml saline (1.0 optical density at 660 nm). The bacteria contained in the S-100 vaccine are listed in Table 1 below.

S-100 Bacteria List Escherichia coli Escherichia coli (Aerobacter) Klebsiella pneumonia Pseudomonas aeruginosa Salmonella typhimurium Salmonella dysenteriae Salmonella enteriditis Salmonella epidermis Salmonella simulans Streptococcus pyogenes, type 1 Purulent Streptococcus, Type 3 Purulent Streptococcus, Type 5 Purulent Streptococcus, Type 8 Purulent Streptococcus, Type 12 Purulent Streptococcus, type 14 Purulent Streptococcus, Type 18 Purulent Streptococcus, type 22 Pseudomonas vulgaris Streptococcus agalactiae Streptococcus mitis Streptococcus mutans Streptococcus salavarius Streptococcus sanguis Streptococcus pneumoniae Propionibacterium acnes Hemophilis influenzae

<Immune Process of Hyperimmune Egg Products>

Preparations of dead pathogens were prepared as described above. For the first vaccination, the bacteria were mixed with Freund's complete immune adjuvant and 5.6 mg of bacterial material was injected into the chicken's pectoral muscles. For the remaining vaccines, bacterial preparations were mixed with Freund's incomplete immune adjuvant and injected into chickens at 2 week intervals for 6 months.

Eggs were collected from the hyperimmunized chicken and spray dried in powder form. During the spray drying process, the injection temperature was not allowed to exceed 160 ° C. (320 ° F), the discharge temperature was maintained so that the final moisture of the prepared powder was in the range of 3.0 to 4.0% and the pump pressure was 2500 to 4000 P.S.I. It was maintained to the extent. Using lower temperatures in the range from 37.8 to 71.1 ° C. (100 to 160 ° F), the samples were monitored for moisture content during the drying process to obtain a final product having any desired concentration.

<Example 2>

Use of hyperimmune egg product in the treatment of diarrhea

The purpose of this test is to identify, characterize and demonstrate the prophylactic activity of hyperimmune egg products in mouse diarrhea test models. In a model using mice, diarrhea was induced by oral administration of castor oil. This test is the standard test for human anti-diabetic agents and was used to find the two most used anti-diabetic agents, loperamide and difeoxylate.

Way

This study was conducted at Product Safety Laboratories, East Brunswick, NJ. Male mice weighing 25-30 g were obtained from Ace Laboratories, Boyertown, PA. They were isolated for at least 5 days and then randomized into 10 groups and placed in plastic boxes with bedding. Two consecutive days from 8:00 to 9:00 A.M. All rats were administered via feeding. Rats were fed 4.0 g / kg orally with powdered immunized or powdered control eggs in 20 ml / kg distilled water. Spray-dried hyperimmune egg powder was obtained from Div. C., Wilmington, Delaware, USA. (DCV, Inc.). This was obtained from eggs of chickens that were repeatedly vaccinated with large amounts of dead intestinal pathogens of human origin. Spray-dried table eggs were used as a control. One hour after the second day of feeding, 0.3 ml of castor oil dose was administered by feeding. The rats were then placed in individual wire-bottom cages, and the stools were observed and recorded 2, 4 and 6 hours after castor oil administration. Positive or negative responses were used to assess diarrhea. The number of normal, smooth or thin sides was also recorded. Stool was assessed as normal or diarrhea in each of these time periods. The results are expressed in the number of rats with diarrhea.

Test A: The following substances were administered to mice.

1.Control egg-DC product A-98-06-Jan ID # 843 lot 951600FB

PSL 80108 3D-4.0 g / kg PO

2. Over-Immune R-DC Product B-98-06-Jan ID # 3059 lot 96298 VOFP

PSL 80108 4D-4.0 g / kg PO

Test B: The following substances were administered to mice.

Distilled water-20 ml / kg PO

2. Over-Immune R-DC Product B-98-06-Jan ID # 3059 lot 96298 VOFP

PSL 80108 4D-1.0 g / kg PO

3. Over-Immune R-DC Product B-98-06-Jan ID # 3059 lot 96298 VOFP

PSL 80108 4D-2.0 g / kg PO

4. Over-Immune R-DC Product B-98-06-Jan ID # 3059 lot 96298 VOFP

PSL 80108 4D-4.0 g / kg PO

Data analysis and statistics

The results were considered to be diarrhea as anal-genital stains and smooth stools with or without all standards. Results are expressed as percentage of rats with diarrhea at each hour and positive cumulative number at 6 hour period. Percent inhibition was calculated as compared to the appropriate control. Statistical analysis using chi square test.

result

The results of the study are shown in Table 2 below. Data is presented as percentage of rats with diarrhea and percentage inhibition relative to controls for studies 1 and 2. Initial studies (test A) comparing hyperimmunized eggs to control eggs showed a significant difference in inhibition of castor oil-induced diarrhea by hyperimmune eggs. Compared with normal eggs, 4 g / kg of hyperimmune eggs significantly prevented castor oil-induced diarrhea in the test subjects during the 4 hour observation period. Diarrhea was not observed in the 2 hour period, and a minimum amount of diarrhea was seen in all groups in the 6 hour period. Significant inhibition in the cumulative calculation of diarrhea over the entire 6 hours was observed in the hyperimmune egg-treated group.

In a second study (test B), three doses of hyperimmune eggs were compared with distilled water. All groups were administered for 2 consecutive days. The study was conducted under conditions similar to test A by administering castor oil on day 2. Minimal diarrhea was observed in the first observation period. However, apparent diarrhea was observed in control treated rats at 4 and 6 hour observation periods. Administration of 1 g / kg of hyperimmune eggs did not show apparent branch activity in any time period. Administration of 2 g / kg hyperimmune eggs showed clear activity in the 6 hour period, but not in the 2 or 0-6 cumulative time periods. Administration of 4 g / kg of hyperimmune eggs had a clear effect on all time periods when diarrhea was observed for the control period. The percentage inhibition of castor oil diarrhea over the 0-6 hour period was dose related, with 38% inhibition at 1 g / kg, 43% at 2 g / kg, and 62% at 4 g / kg. The estimated 50% effect dose was 2 to 4 g / kg.

debate

The hyperimmune egg product showed obvious dose related activity against castor oil induced diarrhea. Oral administration of 4 g / kg of bolus was effective in all studies and was an appropriate therapeutic dose. The mechanism of hyperimmune egg activity on castor oil is unknown but may be related to possible visceral anti-secretory activity. Hyperimmune eggs are not effective in these studies administered by food. This was due to the lack of effective concentration of hyperimmune eggs at the active site.

Hyperimmune eggs will find use as acute therapies that are antidiabetic in both animals and humans. This study demonstrates that long periods of pretreatment to show activity are not necessary and are effective when administered orally in a monoclonal dosage form.

Acute Administration with Castor Oil-Induced Diarrhea-Nutrition Exam A cure Dose g / kg PO Number of mice with diarrhea / number of mice tested 2 hours % 4 hours % 6 hours % 0 to 6 hours % control% Control Egg E80108-4D Egg Protein E80108-4D 44 0/100/10 00 9/102/10 * 9020 3/102/10 3020 12/304/30 * 4013 ---- 68 Exam B cure Dose g / kg PO Number of mice with diarrhea / number of mice tested 2 hours % 4 hours % 6 hours % 0 to 6 hours % control% Vehicle egg protein E80108-4D 0124 1/101/101/100/10 1010100 10/106/107/104/10 ^* 100607040 10/106/104/10 * 2/10 * 100604020 21/3013/3012/308/30 * 70434027 ---- 384362

<Example 3>

Materials and methods

Target person

HIV-positive male subjects ranging in age from 18 to 50 years old and suffering from gastrointestinal disorders (as described in ELISA and Western Blot analysis), Jamshedji Jeejiboy (JJ) Hospital, Mumbai, India Recruited from). These individuals have been previously diagnosed with one or more AIDS (reference) defined as a second state. They did not participate in any study drug trials in the last 60 days or have been exposed to any immunomodulators or vaccines in the last 90 days. Subjects with frequent changes in dosage or type of drug controlling clinical symptoms and who have alcohol or substance abusers, a history of allergies to eggs, or any other change in the test provisions were excluded from the study.

Study design

An open-level study test of food supplements fortified with spray-immunized hyperimmune eggs (DSV, Wilmington, Delaware, USA) was conducted for a total of 12 weeks. Food supplement beverages were freshly prepared and consumed as a liquid once daily for 8 days. At the end of this two month period, the test clause was no longer consumed for the remaining one month of the test. The same physician monitored the subject and took a blood sample at four week intervals throughout the study. All subjects maintained their normal diet and medication and monitored for any appearing signs or symptoms during the 60-day trial period and 30 days thereafter. The severity, date of onset, duration, frequency, study product relationships, actions taken, and the results of each adverse test were recorded.

All subjects had to visit the doctor before starting the study (baseline visit) and then made four visits every four weeks. During each visit, subjects received a general assessment of their well-being with a list of questions including detailed physical examinations, a) evaluating the frequency of bowel movements, and b) consistency of bowel movements.

There were two case reports in the study.

<Case 1>

Subject # 1 weighed 46 kg. He complained of abdominal pain, nausea, vomiting and diarrhea. Treated with fluid administration. For eight weeks of the test period, the subject reported reduced incidence of nausea and diarrhea. The well-being of the subjects was generally improved and he showed weight gain from 46 kg to 47.5 kg (Table 1).

<Case 2>

Subject # 2 weighed 53 kg. He complained of shortness of breath and pain in the abdomen and was treated for Koch's infection. Subjects have been tested positive for HIV for the past six years and sporadically administered ayurvedic drugs for chest congestion and cough. Subjects also received antibiotics and cough syrup.

After consuming the test article for eight weeks, he showed improvement in health and general well-being with a weight gain of 3 kg. Abdominal pain and dyspnea decreased dramatically after 4 weeks of using the product.

Subject's response

Seventeen subjects participated in the study, with 15 (88.2%) gaining weight in the range of 0.5 to 6 kg. In some cases, weight gain increased by 10% from initial weight. The quality of life index assessed by the physician was significantly improved in both the physical and emotional state of the subject. Most appropriately, many subjects also experienced a diminishing or total elimination of diarrhea episodes.

Claims (17)

A method of treating and preventing diarrhea in a subject animal, comprising administering to the subject animal an effective amount of an egg product. The method of claim 1, wherein the subject animal suffers from or is susceptible to diarrhea. The method of claim 1, wherein the egg product is obtained from an egg-producing animal hyperimmunized with an immunoantigen or genetic vaccine. The method of claim 3, wherein the immunoantigen vaccine comprises one or more immunogens selected from the group consisting of bacterial, viral, protozoan, fungal and cellular immune antigens and mixtures thereof. The method of claim 4, wherein the immunogen vaccine is selected from Escherichia coli, Escherichia coli (Aerobacter); Pneumococcal (Klebsiella pneumonia); Pseudomonas aeruginosa; Salmonella typhimurium; Salmonella dysenteriae; Enteritis (Salmonella enteriditis); Salmon epidermis; Salmonella simulans; Streptococcus pyogenes, type 1; Purulent streptococci, type 3; Purulent streptococci, type 5; Purulent streptococcus, type 8; Purulent streptococcus, type 12; Purulent streptococcus, type 14; Purulent streptococcus, type 18; Purulent streptococcus, type 22; Pseudomonas vulgaris; Streptococcus agalactiae; Streptococcus mitis; Streptococcus mutans; Streptococcus salavarius; Streptococcus Sanguis; Streptococcus pneumoniae; Propionibacterium acnes; And one or more immunogens from each bacterial line of Haemophilis influenzae. The genetic vaccine of claim 3, wherein the genetic vaccine is naked DNA, plasmid DNA, viral DNA, bacterial DNA, DNA expression library, DNA-RNA immunoantigen, DNA-protein conjugate and DNA liposome conjugate, and And at least one immunoantigen-encoding DNA construct selected from the group consisting of fragments of the mixture. The method of claim 1, wherein the effective amount of egg product administered to the subject animal is 0.5 to 6 g daily egg product per kilogram of body weight of the subject animal. 8. The method of claim 7, wherein the effective amount of egg product administered to the subject animal is 4 g of egg product per kilogram of body weight of the subject animal. The method of claim 1, wherein the egg product is administered parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, intranasally, orally or topically. A composition comprising a beneficial property of preventing or reducing diarrhea in a subject animal, comprising a product obtained from an animal that is overimmunized with one or more immunogens. The composition of claim 10, wherein the subject animal is suffering from, or prone to diarrhea. The composition of claim 10, wherein the animal further comprises an egg-producing animal. 13. The composition of claim 12, wherein the product further comprises an egg product. The composition of claim 10, wherein the animal is overimmunized with an immunogen or genetic vaccine. The composition of claim 14, wherein the immunoantigen vaccine comprises one or more immunogens selected from the group consisting of bacterial, viral, protozoan, fungal and cellular immune antigens and combinations thereof. The method of claim 15, wherein the immunogen vaccine is E. coli, E. coli (Erobacter); Pneumococcal bacillus; Pseudomonas aeruginosa; Rat typhoid bacteria; Salmonella Daicenteri; Enterococci; Salmon Epidermis; Salmonella Simulans; Purulent streptococci, type 1; Purulent streptococci, type 3; Purulent streptococci, type 5; Purulent streptococcus, type 8; Purulent streptococcus, type 12; Purulent streptococcus, type 14; Purulent streptococcus, type 18; Purulent streptococcus, type 22; Pseudomonas vulgaris; Streptococcus agalactier; Streptococcus mitis; Streptococcus mutans; Streptococcus salavarius; Streptococcus sanguis; Pneumococcal streptococci; Propionibacterium acne; And one or more immunogens from each bacterial line of Haemophilis influenza. The group of claim 14, wherein the genetic vaccine consists of fragments of naked DNA, plasmid DNA, viral DNA, bacterial DNA, DNA expression libraries, DNA-RNA immunoantigens, DNA-protein conjugates and DNA liposome conjugates, and mixtures thereof. A composition comprising at least one immunoantigen-encoding DNA construct selected from.