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WO1997037636A1 - Lutte contre des microbes nocifs: therapie et prevention - Google Patents

Lutte contre des microbes nocifs: therapie et prevention Download PDF

Info

Publication number
WO1997037636A1
WO1997037636A1 PCT/FI1997/000208 FI9700208W WO9737636A1 WO 1997037636 A1 WO1997037636 A1 WO 1997037636A1 FI 9700208 W FI9700208 W FI 9700208W WO 9737636 A1 WO9737636 A1 WO 9737636A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
antibodies
candy
nipple
cream
Prior art date
Application number
PCT/FI1997/000208
Other languages
English (en)
Inventor
Elias Hakalehto
Ilpo Kuronen
Original Assignee
Elias Hakalehto
Ilpo Kuronen
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from FI961492A external-priority patent/FI961492A0/fi
Priority claimed from FI961493A external-priority patent/FI961493A0/fi
Application filed by Elias Hakalehto, Ilpo Kuronen filed Critical Elias Hakalehto
Priority to AU22954/97A priority Critical patent/AU2295497A/en
Publication of WO1997037636A1 publication Critical patent/WO1997037636A1/fr
Priority to PCT/FI1998/000298 priority patent/WO1998043610A1/fr
Priority to ES98912540T priority patent/ES2367696T3/es
Priority to AU67338/98A priority patent/AU6733898A/en
Priority to AT98912540T priority patent/ATE511831T1/de
Priority to EP98912540A priority patent/EP0996423B1/fr
Priority to DK98912540.6T priority patent/DK0996423T3/da

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/02—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from eggs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A61K9/0056—Mouth soluble or dispersible forms; Suckable, eatable, chewable coherent forms; Forms rapidly disintegrating in the mouth; Lozenges; Lollipops; Bite capsules; Baked products; Baits or other oral forms for animals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the object of the invention is therapeutic and preventive method against harmful microbes and substances.
  • Antibodies are proteins produced by the organism and they are found in all more developed animals. The task of antibodies in the organism is to serve as the primary defensive mechanism against unknown components like microbes that penetrate to the organism. Antibodies are consisting of four polypeptide chains that form the functional molecule, which includes two specific identifying and binding sites and a separate frame that contains other biological functions. The most important property of the antibodies is their ability to bind to their target molecules very specifically. This means that with the help of antibodies it is possible to identify and eliminate e.g. only the harmful organisms. The organism defends against the penetrating microbes by forming antibodies that bind to the penetrator.
  • the formation of antibodies in sufficient amounts takes from several days to even weeks, however. During that time diseases may break out. In the course of infection the level of antibodies rises and at last the penetrator will be eliminated. In a similar way it takes a few weeks after the infection is over before the level of antibodies declines to the normal.
  • Antibodies against many kinds of antigens have been produced in hens These include proteins, synthetic proteins, viruses of plants and wall structures of fungal and bacterial cells. Hens have been found to be very effective producers of antibodies A chicken that has been immunized, produces as much antibodies per month as is found in a high titre rabbit antiserum. Specific antibodies can be usually isolated from the egg yolk already after two weeks from the first immunization. The production of antibodies in hen continues at least 100 days. One egg can produce as much as about 10 doses of therapeutic antibodies. The costs per dose is much lower than e.g. in the production of monoclonal antibodies for the same purpose.
  • the microbial material (originating e.g. from bacteria or viruses), which can be inactivated with radioactive irradiation, if required, is injected to the test animals like hens for the production of adequate amounts of antibodies.
  • Antibodies that have been isolated from the egg yolk for different kind of curing, preventive and other probiotic effects can be advantageously purified before their ultimate use. Effective methods 3 for the purification of these antibodies have been developed recently
  • Mucous membranes of respiratory tract provide a very good gateway to the organism for many harmful microbes Air that we breath and food bring along micro-organisms which attach to the mucous membranes of respiratory tract or different parts of alimentary tract from where they penetrate to the organism and cause the actual infection and the outbreak of a disease.
  • the organism tries to defend against the invaders by secreting immunoglobulins A and E (IgA, IgE) to mucous membranes
  • IgA, IgE immunoglobulins A and E
  • the task of the IgA and IgE is to bind to the harmful organisms and by that way to prevent their penetration from mucous membranes to the body or to prevent their binding to the cellular tissue of the mucous membranes, which often precedes microbial infection
  • the mechanism mediated by IgA and IgE is not always enough to protect the body against the organisms coming from outside
  • One reason for this is a so called delay in the mechanism, because the secretion of a large amount of antibodies to the mucous membranes needs a contact with the antigen and requires many days for synthesis During that period of time the amount of micro-organisms inside the body has become adequate for the outbreak of disease In the course of the disease antibodies are secreted to blood circulation where they destroy the invader and also to the mucous membranes where
  • Dental caries for example, has been prevented efficiently in rats by using a therapeutic antibody that has been produced in the egg yolk
  • the purpose of this invention is to introduce a method which can be used for spreading antibodies to the area of mouth and pharynx and other respiratory organs in a efficient and cost-effective way.
  • the exploited antibody or microbe will be transported to the area of mouth and pharynx in a product that will be either sucked or chewed or is in a liquid form.
  • the functioning of a probiotic microbial strain can be promoted by regulating the physical, chemical or biological parameters of mucous membranes or the part of the organism under treatment in a way that the conditions for the probiotic microbe will improve without hampering the normal function of the mucous membranes or the part of the system. It is possible to e.g. add beneficial growth factor for the microbe in question or harmful substances for the competing microbes together with the probiotic microbe.
  • the basic idea of the method of this invention is to get antibodies that have been produced in the egg yolk either in a purified or unpurified form to the mucous membranes of mouth, pharynx and other upper respiratory tract or to the skin surface in a way, that beneficial health effects for human being and test animals will be achieved. In this way the spread of infection in the lungs and ears will be prevented.
  • the antibodies beneficial for health can be placed to candy products in such a form that they are e.g. released when sucking a caramel, biting a chewing gum or in the liquid form liberating slowly from another kind of product, having thus a relatively long-lasting effects on the membranes of the upper respiratory tract.
  • antibodies will dissolve slowly into the mucous membranes of mouth and pharynx and they give a more effective, more meaningful and long-lasting influence. If used repeatedly they can prevent the spreading of microbes through nose/ pharynx tract to the auditory tract.
  • Antibodies produced in eggs can be added to the product e.g. after purification in iyophilized form.
  • the objects can be such as Streptococcus muta s that causes caries, viruses that cause influenza and other infections of upper respiratory tract, and other sources of infections of ears and lungs.
  • the egg yolk antibodies can be made reactive with probiotic microbial strain, which has the ability to bind antibodies through the mannose residue which is situating in the Fc-part.
  • probiotic microbial strain which has the ability to bind antibodies through the mannose residue which is situating in the Fc-part.
  • enteric bacteria with type 1-fimbria bind to this sugar molecule.
  • the same ability can be produced with the help of gene transfer methods by adding the required part of protein to the surface protein of the microbe in use (e.g. flagellin or S-layer protein) Then there will be formed a layer of recognizable antibody onto the surface of the probiotic microbe which helps the probiotic influences to be directed to the desired parts of the organism, cell surfaces, other microbial cells etc.
  • Antibodies produced in the egg yolk can be produced against e.g. influenza, parainfluenza, RSV-, adeno-, rhino-, EBV-, or cytomegaloviruses, rota- or enteroviruses, bacteria that cause inflammation of ear or diseases of the respiratory tract, microbes that cause teeth diseases (e.g. Streptococcus mutans) or other microbes that produce harmful substances in the body
  • These antibodies can be produced also to eliminate influences of toxins and other substances that can be either toxic or harmful (e.g environmental toxins). With the help of this method it is possible to increase the washing out of harmful organisms from the body when they bind themselves through antibodies either to each other or to carrier molecules or to particles or to corresponding substances.
  • They can also prevent the function of the microbe or its reproduction e.g. by preventing transport of molecules through the cell wall, moving or binding of the microbial cells or they can disturb the microbial cell division. They can also aggregate the microbes in a way that they wash out from the body as larger particles or get into places where they are destroyed (e.g. the microbes of the pharynx in the circumstances of low pH in the stomach)
  • Molecules of that kind can be e.g. antibiotics, lectins, components that inhibit or stimulate growth of certain microbes, enzymes, cofactors or other corresponding molecules
  • the binding of these components to the antibodies can be accomplished with different methods either chemically or physically.
  • antibody will be placed in the form of liquid, emulsion or cream on the surface or inside a nipple.
  • a nipple there are paths, clefts, semipermeable surfaces or other corresponding structures through which the liquid, emulsion or cream can seep to the mouth when the nipple is sucked.
  • the material of the mouth-piece of the nipple can also be selected in way that it lets the antibody solution to penetrate and makes thus it possible for it to seep into the mouth With this fashion antibodies can be given even to babies in a reliable and safe way.
  • the liquid that contains the antibody has been advantageously sweetened with a suitable sweetening agent and the liquid is situated inside the nipple.
  • the pH of the liquid can been regulated to the value desired.
  • the nipple can be made for a single use or its base may have a cover that can be opened, and from where the antibody solution can be filled into the nipple
  • Antibodies were produced in a hen (A) by injecting it with a peptide consisting of the 29 N-terminal amino acids of the human parathyroid hormone.
  • the antibodies produced against this peptide were isolated and purified from the egg yolk laid by the hen A according to previously known procedure.
  • the above mentioned antibodies containing product were added to the filled cherry confection (Ginja-che ⁇ y chocolate candy, Regina, Portugal) in a way that firstly the filling of the confection was partially removed with a sterile syringe and a sterile needle Microlance® 3 (length of the needle 50 mm, diameter 1,2 mm) and it was replaced by an antibody solution prepared in the sterile saline, and containing 25 mg of specific antibodies per 0,4 ml of the solution.
  • the weight of the confection was 15,3 g and pH in a crushed form 4,95.
  • the confection was given to the test person to be consumed by smelting it in the mouth. After 5 minutes he was given 10 ml sterile saline solution for rinsing the mouth for 30 seconds and then split to the sampling dish. After mixing a 50 ⁇ l sample was taken, which was mixed in 50 ⁇ l of Assay-buffer. Thereafter the detection was carried out with a peroxidase labeled antichicken antibody (Zymed Inc., CA, USA). An untreated candy and a liquid that was obtained in a similar way as the sample itself was used as a blank sample.
  • a candy containing therapeutic antibody was produced with the following procedure: 200 ml of water was heated to the boiling point and 5 g of agar (E 406) was added. The solution was boiled for 30 minutes to dissolve the agar. Thereafter 14,5 g of sugar, 3,7 g of xylitol and 50 g of cloud berry jam (Kesko, Finland) were added. The solution was froze to 55 °C and poured into a 7 ml mould, after which 25 mg of the antibody according to the example 1 was added in 0,4 ml saline solution. During the addition temperature was about 40 °C and the solution had started to solidify. The pH of the final candy was 4,27.
  • the test person took the candy and after that a sample was collected and analysed after 5 minutes according to the example 1.
  • the blank sample was prepared before the actual sample according to the example 1 and it was analysed in a similar way.
  • Purified and Iyophilized antibody that had been produced specifically against Francisella tularensis in egg yolk according to the example 1, was added to sterile saline solution to the final concentration of the antibody of about 18 mg/ml.
  • Xylitol was added to the solution to the final concentration of 100 mg/ ml.
  • 250 ⁇ l of the solution obtained in this way was placed into the mouth-piece of a nipple, which was incised with a laboratory knife, and the antibody solution was shown to deafened onto the nipple surface through the incisions when the nipple was sucked.
  • the volume of 400 ⁇ l of the antibody solution described in the example 4 was added to 1 ml of Aqualan L (Orion, Helsinki) cream and they were mixed carefully. The emulsion was then spread to the skin and the mucous membranes of mouth. Preservation of the antibody in an active form after spreading was demonstrated with different immunological methods.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Physiology (AREA)
  • Nutrition Science (AREA)
  • Public Health (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

Cette invention a trait à une méthode thérapeutique et préventive des effets nocifs exercés sur la santé par certains microbes et certaines substances. Dans le cadre de cette méthode, on produit un anticorps à partir d'oeufs dont le jaune a été isolé. L'anticorps selon l'invention, qui est spécifique de certains microbes et substances néfastes pour la santé, est placé dans un produit, qui est lui-même introduit dans les cavités buccale et pharyngée, et que l'on peut sucer ou mâchonner, ce produit pouvant encore se présenter sous forme liquide ou bien sous forme de crème ou d'émulsion.
PCT/FI1997/000208 1996-04-03 1997-04-03 Lutte contre des microbes nocifs: therapie et prevention WO1997037636A1 (fr)

Priority Applications (7)

Application Number Priority Date Filing Date Title
AU22954/97A AU2295497A (en) 1996-04-03 1997-04-03 Therapeutic and preventive method against harmful microbes
PCT/FI1998/000298 WO1998043610A1 (fr) 1997-04-03 1998-04-03 Procede de production de produits sucres contenant de la gelee
ES98912540T ES2367696T3 (es) 1997-04-03 1998-04-03 Un método para producir dulces de gelatina que contienen anticuerpos.
AU67338/98A AU6733898A (en) 1997-04-03 1998-04-03 A method for producing jelly sweets which contain antibodies
AT98912540T ATE511831T1 (de) 1997-04-03 1998-04-03 Verfahren zur herstellung von antikörper enthaltenden gummibärchen
EP98912540A EP0996423B1 (fr) 1997-04-03 1998-04-03 Procede de production de produits sucres contenant de la gelee
DK98912540.6T DK0996423T3 (da) 1997-04-03 1998-04-03 Fremgangsmåde til fremstilling geléprodukter indeholdende antistoffer

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
FI961492A FI961492A0 (fi) 1996-04-03 1996-04-03 Terapeutiskt och profylaktiskt foerfarande foer bekaempande av skadliga mikrober och aemnen
FI961493 1996-04-03
FI961493A FI961493A0 (fi) 1996-04-03 1996-04-03 Foerfarande foer riktning av probiotiska verkningar
FI961492 1996-04-03

Publications (1)

Publication Number Publication Date
WO1997037636A1 true WO1997037636A1 (fr) 1997-10-16

Family

ID=26160143

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FI1997/000208 WO1997037636A1 (fr) 1996-04-03 1997-04-03 Lutte contre des microbes nocifs: therapie et prevention

Country Status (2)

Country Link
AU (1) AU2295497A (fr)
WO (1) WO1997037636A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002002642A3 (fr) * 2000-07-05 2002-04-18 Bergter Wolfgang Composition pour des produits pharmaceutiques et cosmetiques
WO2005005481A3 (fr) * 2003-07-10 2005-04-28 Avitek Pharma Inc Therapie combinatoire pour maladies gastro-enteriques provoquees par des micro-organismes
EP2394627A1 (fr) 2010-06-14 2011-12-14 Nestec S.A. Dispositif d'alimentation
EP2441432A1 (fr) 2010-10-13 2012-04-18 Nestec S.A. Dispositif d'aspiration avec dépôt externe de probiotiques
US11241030B2 (en) * 2011-10-06 2022-02-08 Societe Des Produits Nestle S.A. Edible web comprising microorganisms

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993015736A1 (fr) * 1992-02-18 1993-08-19 Pharmos Corp. Compositions seches pour la preparation d'emulsions submicroniques
DE4324859A1 (de) * 1992-07-23 1994-01-27 Lion Corp Oral verabreichbare Zusammensetzung zur Behandlung periodontaler Erkrankungen
WO1994021284A1 (fr) * 1993-03-15 1994-09-29 Pharma Pacific Pty. Ltd. Formulation et methode therapeutiques

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993015736A1 (fr) * 1992-02-18 1993-08-19 Pharmos Corp. Compositions seches pour la preparation d'emulsions submicroniques
DE4324859A1 (de) * 1992-07-23 1994-01-27 Lion Corp Oral verabreichbare Zusammensetzung zur Behandlung periodontaler Erkrankungen
WO1994021284A1 (fr) * 1993-03-15 1994-09-29 Pharma Pacific Pty. Ltd. Formulation et methode therapeutiques

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DIALOG INFORMATION SERVICES, File 351, DERWENT WPI, Dialog Accession No. 008015737, WPI Accession No. 89-280849/198939, GEN CORP KK et al., "Antibody Used to Prevent Dental Caries - Where Immunogloculin Fraction Isolated From Egg Yolk Inhibits Streptococcus Mutans From Producing Plaque"; & JP,A,01 203 317, *
DIALOG INFORMATION SERVICES, File 351, DERWENT WPI, Dialog Accession No. 008062519, WPI Accession No. 89-327631/198945, GEN CORPORATION KK, "Antibody, for Preventive Agents for Cariogenic Teeth - Comprises Immunoglobulin Prepd. from Eggs of Chickens Immunised with Water-Insol. Glucan Synthetic Enzyme"; & *
DIALOG INFORMATION SERVICES, File 351, DERWENT WPI, Dialog Accession No. 008958579, WPI Accession No. 92-085848/199211, TAKARA SHUZO CO LTD, "Skin Cosmetic with Prolonged Effect - Contains (Modified) Antibody and Usual Carrier"; & JP,A,04 029 912, (31-01-1992). *
DIALOG INFORMATION SERVICES, File 351, DERWENT WPI, Dialog Accession no. 011162916, WPI Accession No. 97-140841/199713, LOTTE CO LTD, "Chewing Gum to Prevent e.g. Oral Infection - Contains Egg Antibody Added to Chemical Gum Base"; & JP,A,09 020 684, (21-07-1997). *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002002642A3 (fr) * 2000-07-05 2002-04-18 Bergter Wolfgang Composition pour des produits pharmaceutiques et cosmetiques
WO2005005481A3 (fr) * 2003-07-10 2005-04-28 Avitek Pharma Inc Therapie combinatoire pour maladies gastro-enteriques provoquees par des micro-organismes
EP2394627A1 (fr) 2010-06-14 2011-12-14 Nestec S.A. Dispositif d'alimentation
WO2011157532A1 (fr) 2010-06-14 2011-12-22 Nestec S.A. Dispositif d'alimentation
CN102939068A (zh) * 2010-06-14 2013-02-20 雀巢产品技术援助有限公司 进给装置
EP2441432A1 (fr) 2010-10-13 2012-04-18 Nestec S.A. Dispositif d'aspiration avec dépôt externe de probiotiques
WO2012048914A1 (fr) 2010-10-13 2012-04-19 Nestec S.A. Dispositif d'aspiration
CN103153262A (zh) * 2010-10-13 2013-06-12 雀巢产品技术援助有限公司 吸吮装置
US20130200030A1 (en) * 2010-10-13 2013-08-08 Nestec S.A. Suction device
AU2011316082B2 (en) * 2010-10-13 2015-04-02 Nestec S.A. Suction device
RU2571498C2 (ru) * 2010-10-13 2015-12-20 Нестек С.А. Сосательное устройство
US11241030B2 (en) * 2011-10-06 2022-02-08 Societe Des Produits Nestle S.A. Edible web comprising microorganisms

Also Published As

Publication number Publication date
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