IL85963A - Pharmaceutical compositions for the prophylaxis or treatment of disorders or diseases of the gastrointestinal tract, comprising antigenic substances - Google Patents

Abstract

Disorders and diseases in human and animal digestive systems, for example diarrhoea, ulcerative disorders and carcinomas, are a serious, economically important problem worldwide. It was possible to show that immunological mechanisms exhibit a protective effect in the gastrointestinal tract of animals and humans. Antigens contained in the food induce the said protective mechanisms if the body has previously been sensitised to this antigen. The use of antigenic substances in the prophylaxis and therapy of ulcerative disorders in humans is preferred. The antigenic substances according to the invention are furthermore particularly highly suitable in the raising of young cattle, which are subjected to a change of food in the first weeks of life. <IMAGE> [EP0286847A2]

Description

85963/4 nn>"TO >N !?io>oi ny»jot? mnpn >->>¾on >JA>OJN lam o>^>Dan .inD'yn ro~>ya v ni^nni Pharmaceutical compositions for the prophylaxis or treatment of disorders or diseases of the gastrointestinal tract, comprising antigenic substance Effem GmbH P 20 824 - 002/ja March 14, 1988 Prophylaxi s or a the rapy of disorders and di seases within the gastro ntest nal tract of animals and humans using antigenic substances D E S C R I P T I O N The invention refers to the prophylaxis and therapy of disorders and diseases within the gastrointestinal tract of animals and humans using an antigenic substance.

Horbid disorders with n the gastrointestinal tract of animals and humans represent a worldw de serious, economically s gnificant problem. In human medicine, among chronic disease, peptic ulcer disease, for instance, shows a life long prevalence of about 8 - 10% in Europe and North America. Thus, the total costs for peptic ulcer patients in U.S.A., The Netherlands, Italy and Sweden, including working deficit are about 1% of the respective national annual expenses for health care. In Germany, for instance, the mortality because of peptic ulcer disease within the period from 1952 to 1980 has decreased only i nsi gi f i cant ly from 7 to 6 deaths per 100,000 inhabitants (Sonnenberg R., Fritsch, A.: Changing mortality of peptic ulcer disease in Germany. Gastroenterology 84, 1553 (1983)). Within this period, however, peptic ulcer disease, as a deadly disease of middle aged and elderly people, shifted to disease of mainly old and high-aged people. Besides the mortality because of gastric and duodenal ulcers, in women, has increased. The mortality in men because of a duodenal ulcer remained constant; those of gastric ulcers slightly decreased.

Currently/ mainly H2-ant agoni st s are used for the treatment of peptic ulcer disease (Bauernfeind et al., U lcustherap e, JAMA, Heft 2, 135 (1986)). The principle of the therapie with H2-antagonists is ba.s.ed on the reduction gastrointestinal tract. This therapy is effected with a number of unpleasant side-effects. A significant problem of a peptic ulcer disease sti ll is the prophylaxis of a recurrent ulcer because, as already mentioned/ peptic ulcer disease is among the chronic diseases with life-long prevalence. For instance/ a long term study of three years among 44 Centres w th n 13 countries with a total of 1,423 patients yielded that only 81% of patients were without recurrence af ter 12 months, 73% after 24 months and 65% after 36 months for duodenal ulcers (DMW 111. Jg. Nr. 3, 117, (1986)) . Therefore, there is the urgent necessity for alternative drugs for the prophylaxis or treatment of peptic ulcers thout side effects which especially could be taken for a long time and as prophylaxis. 0 ί arrhoaea and peptic ulcer disease are a serious problem not only in human medicine but also in veterinary medicine, especially in pig and cattle breeding as well as in poultry f arming. It is a well known phenomenon, for instance, that p glets after weaning and a change to semi -liquid igh energy ratio wi ll develop ulcers which may partially be lethal. Within the Common Market there are 120 to 160,000,000 pigs (estimated values), among which three to four percent wi ll be affected. Based upon these f igures, the loss of pigs will account for about 4 mi llion per year, resulting in a financial loss of up to 1, 2 billion German Marks per year. It is obvious that effective measures against these disorders are necessary. Nearly the same is true for cattle breeding and poultry farming.

For the pathogeny for peptic uLcer disease, aggressive factors during digestion, like the secretion of acid and pepsin, seem to be of special significance (Grossman, H.I.: Regulation of gastric acid secretion. In: Physiology of the Gastrointestinal Tract. Ed.: L.R. Johnson. Raven Press, New York 659-671 (1981)).

Opposite to these aggressive factors are protective physiologic digestive mechanisms. Among these protective mechanisms is the mucus secretion in the stomach, which together with the bicarbonate secretion is regarded as the first layer of a mucosal barrier.

The various secretion components of the stomach, as -well as the cell regeneration, are controlled by neurohormone I mechanisms. The hormone, which initiates the above-ment oned aggressive digestive processes is gastrin. It was isolated in 1964 (Gregory, R.A., Tracy, H.J.: The constitution and properties of two gastrins extracted from hog antral mucosa. Gut 5, 103-117, (1964)).

Until recently, one aspect concerning studies on the stimulation of gastric functions was totally not taken into account: with each meal a mucosa of the stomach also comes into contact with substances which might be considered antigenic. Although there were hints for immunological structures in the stomach, however, there was no idea as to their functions. Recently, it could be shown, in experimental studies, that following preceding immunization with an antigen, oral challenge with said antigen induces an immune reaction in the antrum of the stomach, which initiates the complete gastric cascade of digestion. This is mainly the increase of local blood flow - - in the antrum and duodenum, increase of mucus secretion of the stomach as well as stimulation of gastrin release.

FR-A-2408351 corresponding to DE-OS-2849226 disclosed a method for improving growth of poultry by administering orally subclinic doses of coccidia-oocytes. In the distal intestinum, the antigenic spores are released from the oocytes and a local immune response of the distal intestinal occurs.

Proof of an immunological stimulation of gastric functions which offers a completely new view of physiology and pathophysiology of the digestion came initially from data resulting from animal studies in which luminal antigens, i.e. Human Gamma Globulin (HGG), in the stomach, after preceding immunization were recognized and induced digestive processes (Teichmann, R.K., Andress, H.J., Gycha, S., Seifert J., Brendel, W.: Immunological mediated gastrin release.

Gastroenterology 84,2 (1983)).

Immunof luoresence microscopy revealed binding sites for the antigen applied orally within the lamina propria mucosa of immunized animals. Furthermore, there was an increase in the number of cells expressing immune associated antigens on thei r surface (so-called la-antigens) in the antrum, following antigen admini stration. Using' double immunof luoresence microscopy it could be demonstrated that the binding sites for the antigen seems to be located on la-antigen expressing cells.

Furthermore, there was an increase in plasma in mast cells, in the lamina propria mucosa of immunized animals, after antigen administration and an increase in the mast cell d eg r a nu I a t i on, as well as an increase in intraepithelial lymphoid cells. (Teichmann, .K., Andress, H.J ., Liebich, H., Seifert, J., Brendel, W. : Die Bedeutung immunkompetenter Zellen im Antrum bei der Stimulation von Verdauungsprozessen. Langenbecks Arch. Chi r. Suppl. 151, (1984)).

Thus, during i mmuno L og i c a L L y mediated stimulation of gastric digestive processes, besides agressive factors Like acid also zytoprotective mechanisms Like increase of bLood fLow and mucus secretion, are initiated.

Based upon these resuLts further experiments were carried out concerning mediators of the mmunoLogi ca L stimuLation of gastric functions using the modeL of isolated ant rum-muscu L a r perfusion in dogs. Gastrin and prostagland ne and ?2 determinations were performed in an intravenous effluent.

The supernatant of fluoresic duct Lymph, collected after antigen administration in immunized animals, stimulated gastrin release n isolated antrum perfusion. This activity was missing in the Lymph of non-immunized animaLs when the same antigen was administrated into the stomach, (Teichmann, R.K., Pratschke, E., Grab, J., Tutert, J., Enders, G., BrendeL, W.: G as t r i n s t i mu L e rend e Wirkung von Ductus-thoraci cus-Lymphe nach ga s t ro i nt e s t i na I e r I mmun re a k t i on . Langenbecks Arch. Chir. Suppl. 271 (1985 ) ) .

Thus, it has been shown that soluble mediators are responsible for the antigen induced gastrin release.

The mucosa mast cell products Histamin and Leukotrien C^, being released during the immunological reactions, significantl inhibit antral gastrin release. Gastrin itself stimulates the release of Histamin so that these results, for the first time, show local antral negative feed-back mechanism between the G-celL and the mast cell (Pratschke, E., Teichmann, R.K., Grab, J., Tutert, E., Brendel, W.: Der EinfluR von MastzeL lprodukten auf die Gast ri nf rei setzung. Langenbecks Arch. Chir. Suppl., 295, (1985) ) .

Simultaneous to the inhibition of gastrin release by Histamin and Leukotrien C^, there is a release of endogenous prostagLand ne Έ.^ and Fg. In the modeL of isolated antrum perfusion it could be shown that prostaglapdine E£ inhibits the gastrin release. This might indicate that the inhibition of gastrin, by mucosa mast cell products, may be mediated by endogenousLy released prostagl andi nes (Pratschke, E., Teichmann, R.K., Grab, J., Enders, G., Brendel, W.: Endogenes P ros t ag L and i n E2 aLs ReguLator der Gast ri nf rei setzung. In: Kong reflbe ri cht der 26. Jahrestagung der osterr.

Gesellschaft fur Chirurgie. Hrsg.: F. Helmer, E. Horcher, Styria-Verlag, Graz, 131, (1985)).

During the process of antigen recognition involving antigen presenting cells lymphokines Interleukin 2 and Gamma interferon are released. Both stimulate gastrin.

Thus, for the first time, stimulation of a gastrin by mediators of immuno competent celLs was shown (Pratschke, E., Teichmann, R.K., Grab, J., Hammer, C. Brendel, W . : Gast ri nf rei setzung durch Mediatoren i tnmunk ompe t ent e r Zellen. Langenbecks Arch. Chir. Suppl., 261 (1986)).

Immunof Luoresence microscopic studies of the antrum of the perfusion with Interleukin 2 and Gamma interferon, using monoclonal antibodies against class 2-antigens of the HC complex, revealed further that, after the perfusion with antigenic structure of DLA Class 2 ( og-Leukocyte antigens), same are expressed on ep thelial cells, macrophages, probably also on intraepitheLial T-Lymphocyte. There was no expression of those two antigens after perfusion with Interleukin 2.

Therefore, Interleukin 2, but also Pros t ag I and i nes and other substances, may serve as mediators of this immunological reaction.

Thus, the above-mentioned antigen induced reactions are discussed for the development of ulcers and may also be of significance for the total gastrointestinal tract.

Whenever antigen induced increase of the antrum blood flow, gastrin and mucus secretion is missing, or disturbed locally, the resulting acid may not be sufficiently buffered and transported back in case of lack of increase of blood flow, resulting in penetration of the acid into the mucosa and possibly inducing local ulcers there.

In est gations on the immunolog cal stimulation of gastric reactions also showed that after immunological stimulation with a synthetic antigen, besides the antigens specific in view of the gastrin release, and the strongest stimulator for the gastr c acid secretion, in the sense of an increase of aggress ve digestive processes, also an increase of antral secretion of Somatostatin was noted which has a protective effect by inhibition of acid and pepsinogen secretion, (Kramling, H.-J., Teichmann .K., Merkle, R., Enders, G., Brendel, W . : Gastrale Protektion durch mmunolog sch induz erte Somatostatinfreisetzung, 153. Tagung Vereinigung N i ed e r r he i n i s c h-We s t f a I i s c h e r Chirurgen, Koln, 9.-11. Okt. 1986).

This data results in a completely new far reaching, and so far not recognized, possibility to specifically influence digestive processing as a protective mechanism concerning all disorders and diseases within the gastrointestinal tract. In human medicine, diseases like erosions, peptic ulcers and, last but not least, cancer are of greatest interests. In veterinary medicine interest is mainly in the critical transition phase of young animals following weaning and subsequent contact with high energy food because of occurrence of erosions and peptic ulcers dur ng this phase.

There is a protective induction of functions of the ?as sfiieRiS!tiiR8ijaaist6nb«fiidry 9Si$hi functions and protective physioLogic digestive processes, which may be used in a broadest sense; this is the use of an antigenic substance for the prophylaxis and therapy of disorders and diseases of the gastrointestinal tract of animals and humans inducing protective physiological digestive processes after an oral challenge of the animal or human with the antigenic substance following preceding i mmuni zati on.

When applying an artj_q e n i c_ s u b s t an.ce , as outlined by the invention/ the stimulated gastric functions may work as follo s: After immunization of the stomach with an antigen which is present in the food the subsequent contact of this antigen, in the food, with the gastric mucosa, the antigen ll be recognized by the so-called antigen presenting cells, for instance, macrophages and presented to the T- I ymphocytes by the help of class 2 antigens. An indication for a' lympocyte mediator immunological stimulation of the above-mentioned gastric functions was demonstrated in an experiment in which immunized dogs were i rradiated by an 80 Rad of an ultra-hard X-ray resulting in an inhibition of the immunologically stimulation of gastrin release based upon destruction of lymphocytes. The capabi lity of the gastrin cell to be stimulated, however, is not effected by the i rradiation.

Following the contact of the antigen with the stomach, being immunized as outlined above, there is a type of cascade of release of Interleukin 1 and stimulation of T-Helper-Lymphocytes. The Helper cells themselves will release Interleukin 2 and Gamma interferon, two I ymophok ί nes which were shown to be able to release gastrin. A significant protective physiological mechanism/ ■namely the increase in mucosa blood flow representing one of several steps of the cascade of protective digestive processes, may be caused by mediators of mast cells, especially Histamine. Besides the expression of class-2-antigens on antigen presenting cells of the antrum, Gamma interferon also induced the expression of these antigens on epithel cells. Possibly this additional recruitment of epithel cells represents a potentiation of antigen recognition. The vagus nerve interferes th this antigen recognition system, in the sense of i mmu nomod u I a t i on . The mentioned inter-relations are shown in Figure 1.

The ulcer protective effect of antigen specific stimulation of digestion has been demonstrated in an animal model. Using the model of the "SHAY"-Ulcer (Shay. H., Komarov, S.A., Fels, S.S., Mekanze, D., Gruenstein, M., Siplet, H. : A simple method for the uniform production of gastric ulceration in the rat. Gastroenterology 5, 43, (1.945 )), the influence of antigen stimulated regulation of the digestion was investigated. Male Wistar-Rats were immunized systemically with a Hapten. Afterwards, the Hapten was administered into the stomach and as control sodium chloride was given. In further control studies, the Hapten or sodium chloride, respect vely was also administered in non-i munized animals. These rats- being treated in such a way were investigated macroscopi cal ly and microscopically to check gastric lesions after their death but not later than 18 hours. There was a sign ficantly less expression or number of perforations and Large wall necroses in the core vent ri cuLous in the group of animals which had been immunized with the Hapten (Figure 3.).

Thus, using the model of the "SHAY-Ulcer" the protective effect of an antigen specific stimulation of the digestion could be demonstrated. Although there is an ulcerogenic effect by release of gastrin and, consecuti ely of acid, as an aggressive factor, simultaneously there is also st mulat on of protective mechanisms, for instance, an increase in blood flow, mucus secretion, possibly through release of Somatostatin, P ros t ag I and i nes and other mediators which finally prevent the occurance of severe secretions of the gastric mucosa or gastric wall. This ulcer protective effect of an antigen nduced regulation of the digestion, as shown here, may t e I eo I og c a 11 y well be regarded as a "log cal" regulation w thin physiological processes, as it results simultaneousl in a stimulation of the digestion and a reduct on of the risk to develop an ulcer. Thus, the develoment of peptic ulcer may well be understood as a local failure of the stomach to be ready for antigenic react ons beside other known causes. The stimulation of gastr c immunological mechanisms by immunization with antigens, therefore, results in a completely new possib lity for the prophylaxis and therapy of disorders and diseases within the gastrin intestinal tract of animals and humans offering broadest indications. The essential, other advantage of use of antigenic substances, for this purpose, is based on the fact that the antigens being used are biological substances with which the mammal as well as the human being comes into contact naturally and, thus, there is no aggressive effect on physiological processes but, moreover, an induction of a logical protective physiological process in a natural manner and, thus, same is without side effects.

In respect of the mentioned mechanisms of the immunological stimulation of the digestion, it is, of course, of special advantage to use such substances as antigens which later will also be included within food. Following immunization of the animal or the human with the antigenic substance a level of factors, within protective effect, will be increased in such a way that disorders or diseases of the gastrointestinal tract will be prevented or treated. Thus, plant proteins, preferrably from maize or soya bean, but also animal proteins, are especially suitable for the immunization because these proteins are often found in food or food products of animals or humans.

Whey prote n can, for example, be used as an amimal protein. It became apparent that the prophylactic influencing of gastral mucosal changes (erosions), e.g. in pigs, which were additionally administered whey protein via the drinking water during the entire fattening period until slaughtering of the pigs after a fattening of 10 weeks, entailed that in particular serious changes of the mucous membrane in the animals treated, as described above, occurred less frequently by 70% than in a control group. The whey protein should preferably be present in a 3 to 9% whey protein solution and especially preferably in a 6% whey protein solution.

Human proteins, which are components of blood or which may be found in serum or other blood derivates, for instance, hemoglobin are suitable to be used as antigenic substances in the sense of the invention.

Also other antigens, which do not necessarily correspond with substances being present in the ingested food, may be suitable for the induct on of the immunological situation which i s responsible for the above-mentioned induction of mechanisms. For instance, in case of immunization with proteins derived f rom the envelopes of vi rus, an immunization may be possible, which wi ll work in the sense of the above-described stimulation of the digestion, but also may cause an ""' "rm munization against virus.

Same is true for antigenic substances which are present in microorganisms with which the gastrointestinal tract may come into contact in one way or the other. The stomach might induce the above-mentioned protective processes as well. Specially suitable seem to be lipopolysaccharides of Escherichia coli, a natural commensal bacteria of some mammals whi ch is always found in the food as contam nation and, thus, seems to be suitable. Another suitable antigenic substance is any component of a microorganism being a member of the protozoae .

The use of an antigenic substance being present as a synthetically-produced protein, preferably as a short peptide which contains antigenic determinants for the induction of an immune response, is especially preferred because then a special and di rectly applicable prophylaxis and therapy will be possible.

A well suitable substance for the induction of the immune situation to induce the protective mechanisms are Haptens., preferably nitrophenyl acetic acid (NIP). This substance has shown its special usefulness in the experiment to induce the immunization, which prevents the development of mucosal lesions after intraluminal challenge of an antigen specific stimulation of the stomach. Especially with this substance, an mmunologically mediated ulcer protection was shown for the f rst time.

Preferably, the above-mentioned nitrophenyl acetic acid is used by coupling same to a carrier protein, namely ova I bumi n.

As outlined in the description above, immune modulatory substances are in the equilibrium state between the aggressive and protective digestive mechanisms. The addition of immune modulatory substances together with the antigenic substances may, therefore, cause an increased effect of the induced protective digestive mechanisms.

The use of combinations of one or several of the above-mentioned substances may also result in an improved prophylaxis or therapy.

T h e . pha rma c e u t i c a I confect oning of one or several of the above-mentioned ant genic substances, perhaps by adding of immune modulators, may be performed for the enteral as well as for the parenteral application. Concern ng the enteral confection ng, aerosols, being applied to the nose or the bronchial system, are preferably used. The confection ng of the parenteral application of antigenic substances preferably concerns substances to be used for ntramuscular and/or sub-cutanous applications.

In the veterinary medicine, a preferred use of the antigenic substances is to immunize piglets with the antigenic substance about two weeks after birth. As has already been mentioned above, piglets suffer from peptic ulcers after the weaning and the change to semi-liquids energy-food. They may even die from these ulcers. Treating these p glets in this critical phase with antigens which later will be present in the food prevents, or at least, reduces damages. The proteins being used as antigenic substances are biological substances which, after application, wil l_b.e__m.e-t.a.b.oJ.J_zed_ complete I y without residue or being excreted in only small amounts.

An especially preferred treatment of the piglets is to immunize them ntramuscularly/ preferably the first injection being about two weeks after birth; a second injection about four to five weeks after birth and a third injection shortly before changing of food. In respect of their practicability/ the animals could then be immunized when they receive iron-injections during breeding. The antigenic substances may be packed in a kit which already contains ron.

When the piglets receive the food which contains said prote ns, with which the animals were immunized, the antigen may also be given to the drinking water in order to further induce and maintain the protection of the gastric mucosa by the described immunological mechan sms.

In human medicine, the use of antigenic substances is preferably suitable to prevent or reduce erosions, peptic ulcers or cancer in the human gastro ntestinal tract. The ncreased production of mucos may prevent the mucosa of oncogenic substances. Furthermore, oncogenic substances, which are known for the organism, may become inaffected by the induction of the digestion.

Also, there is a new preferred and broad spectrum of ind cations in the prophylax s concerning diarrhoea in travellers who come into contact with food products during their trip which are, so far, unknown by their gastrointestinal tract, this means that the gastrointestinal tract, so far, had not been in contact with such food products. The lack of induction of digestion may cause passage of protein into deeper parts of the gut with subsequent* possi bLe overgrowth of bacteria.

It is also a well known problem in the breeding of domestic animals that following transportation of the domest c animals to places at which the breeding will be continued after the first week of Life, the changes in the environment and food may cause significant diarrhoea which, again, may be the cause of other bacterial or viral diseases of the whole ga st roi nt ens t i na I tract.

Immunization of these animals with antigenic substances, being known to be present later in the food, may prevent these disorders and diseases.

One possible use of an antigenic substance is to sens tize p glets parenterally as of the third week at intervals of fourteen days with 2 ml each of the 6% whey protein solution without the addition of adjuvant that is to say on the whole with 4 ml. After weaning of the piglets in the second week and gradual change over from pig breeding feed to coarse grain fattening fodder the piglets were brought into another pig sty and kept in groups in loose housing boxes. From this time on approximately as of the tenth week, all pigs were exclusively fed with very finely ground fattening feed. In literature the admin stration of such feed is indicated, besides stress factors, as the decisive cause of the formation of mucosal changes, such as hyperkeratoses, erosions or ulcera in the gastric mucous membrane of the pig.

The pigs were administered 3 to 6% whey protein with the drinking water during the entire fattening period until the slaughtering of the pigs, after a fattening of ten weeks.

In animals which have been treated in this way changes of the mucosa-like erosions appeared much less frequently.

In the same way as the antigenic substances have been used for the treatment of piglets, calves may be immunized parenterally and/or enterally in their critical phases of life. It is preferred to give intramuscular injections of the antigen in calves within about the first four weeks after birth, a second injection at about the sixth week and a third injection in about the eighth to tenth week when changing to calf food.

Within the first weeks of breeding, the calves receive milk and calf food and, generally, energy food from the eighth to tenth week. From the time of changing to the high-energy food, the antigenic substance is added, preferably to the food and/or drinking water to use and/or maintain the level for the protection of the intestinal tract by the described immunolog cal mechani sms.

The immunization is preferably performed also with adult ruminents. Bulls and cows increasingly suffer from gastric ulcers of different stages during the high lactation period. These ulcers are also especially considered as the cause of subsequent infectious diseases of the whole gastrointestinal tract in adult cattle. Enteral or parenteral treatment of animals with the antigenic substances may prevent these damages.

Another field of using the antigenic substances is the treatment of domestic poultry, for instance, chicken, turkeys, geese, ducks and turkey-cocks. The major sources of food, for the breeding of domestic poultry, are grain granules, fish flour, soya and grinded peanuts. To induce the unisation, in respect of the above-mentioned food substances, a parenteral, injection of the antigenic substance wilL be performed on the first day after hatching because of pracical reasons. Later, immunizat ons for maintenance of a preceding immunization may be performed by application of the antigen via the drinking water within the earLy days or weeks of life.

The current invention will now be described, in more detail, in the following Figures and Examples.

It s shown i n (F_i gu_r_e _1 : The regulation of antigen stimulated gastrin release.

Luminal antigen is recognized by antigen presenting cells and may stimulate mast cells as well as T- I mphoc y t e s among others. The T-cell products Gamma interferon and Interleukin 2 cause release of gastrin which, in turn, stimulates Histamin. The mast cell products Histamin, Leukotrien and P ros t ag I and i ne again inhibit the gastrin release (from negative feedback). The vagus nerve may also influence the T-cell systems and the antigen presenting cells. gu_r_e _2 : Ulcer index using the model of the alcohol ulcer of the rat .

Rats being immunized with NIP show a sign f cantly lower ulcer index following intragastric application of the antigen.

Fj_g_u r e_ 3 : Percentage of animals with lesions, induced by the SHAY-Ulcer model in controls and in animals immunized with NIP-ovalbumi n.

The intragastric application of NIP-human Gamma-globin (NIP-HGG) results in a significantly lower expression of peptic ulcers in immunized animals.

F j_3_u_r e_ : Quantitative changes of the e valuation scales (0 and +++) after a pathological-anatomic evaluation of the gastric mucous membrane of pigs in th e area of pars proventri culari s after a 10 w eek feeding of very f inely ground feed rations (%) .

E x_a_mjD_L e 1 The purpose of the described examples was to investigate the effect of the immunological processes on the other induction concerning stimulation or protection.

Male Wistar-Rats (250-350g) were immunized systemically with NIP-OA (Nitrophenyl acetic ac id-Ova I bum i n ) . To induce ulcers, 1 ml of absolute alcohol was administered into the stomach under ether anesthesia. Fifteen minutes prior to the alcohol application, one group (I) of the animals (n=12), received the antigen NIP (coupLed to HGG as carrier-protein), Group II (n=12), the carrier protein HGG aLone. Non-immunized animaLS six having received NIP-HGG (III), a further six HGG (IV), proved as further controls. An hour after the adm n stration of alcohol, the lesions were determined after the animal was killed and the stomach removed. The lesions were measured mac roscopi cal ly by estimatation of the length of the hemorrhagic lesions.

Animals having been immunised systemically w th NIP developed significantly less mucosal lesions after oral NIP-application and. subsequent administration of alcohol (p = 0,02) than all control groups.

Tab_l_e_1 Immunisation Intragastric Length of Administration of hemorrhagic Les ons I (n=12) NIP NIP 24 p=0,02 II (n=12) NIP HGG 57 III (n=6) non-i mm. NIP 72 IV (n=6) non- i mm . HGG 78 The intraluminal antigen specific st mulation of the stomach in immunised Wistar-Rats significantly reduced the development of mucosal lesions. Therefore, for the first time, an immunologically mediated ulcer protection is shown (Figure 2).

E x_amj3 L e__2 The immunization of male Wistar-Rats (250-350g) was performed using NIP-OA (Nitrophenyl acetic a c i d-Ova I bumi n) . Pylorus ligated rats (SHAY-Ulcer) served as uLcer model. Following ligation of the pylorus under c h I ora I hyd rat e anesthesia, the immunized rats received the antigen NIP (coupled to human gamma globulin) directly into the stomach (test group). The control animals received only the carrier protein HGG. Another control group consisted of sham operated animals (without pylorus ligation). Eighteen hours after the operation, the changes in the stomach wall were investigated macroscopically and m croscopically. The lesions were defined as follows: Perforations, confluent transmural ulcerus and all necroses of more than 0,5 cm in diameter.

Animals which were systemically immunized with NIP showed significantly less lesions after pylorus ligation and intragastric application of NIP-HGG (p = 0,05) than controls which had received HGG. Sham-operated rats showed' no mucosal lesions.

Table 2 Intragastric Proportion of Application of Animals showi Lesions Test group (n=24) · NIP 7/24 Control group (n=23) HGG 14/23 S h a m - o p e r a t e d (n=6 NIP resp. HGG 0/6 resp. r e s p .6 ).

The antigen specif ic stimulation of gastric functions result in a signif icant reduction of wall lesions in a def ined ulcer model (Pylorus ligation) (F gure 3.).

Ex_a mp_l_e _3 PjjD_c_e d u_r_e_ _ g f I_m m u jvi_z a t_i_o n _ j_ _P i c|_l_e t s Supposedly the antibody production in piglets starts f rom the second week and regarding the practicability, the piglets are immunized and receive i ron injections during thei r breeding by the breeder.

A biologic protein or a synthetic antigenic immune stimulating substance wi ll be used as antigen. It wi ll be metabolized without residues following digestion by the piglets or it wi ll be excreted, as such, in only small amounts.

For the present immunization of the pigLets whey proteins, namely as a 6% whey protein solution/ were adm n stered to the piglets without the addition of an adjuvant or during the later course of immunization as a 3 to 6% addition to the drinking water.

Th rty pigs were taken as a test group and thirty pigs as control animals for an examinat on of the prophylactic effect of the whey proteins administered as antigens regarding gastral mucosal changes. During the test five pigs of the test group and two of the control an mals died as piglets or during change-over.

The test animals were pretreated according to the follow ng scheme. Piglets were sensitized parenterally as of the third week at intervals of 14 days with 2 ml each of a 6% whey prote n solution without the^ addition of adjuvant that is to say on the whole with 4 ml.

After weaning of the piglets in the second week and gradual change over from pig breeding feed to coarse gra n fattening fodder both the test and the control animals were brought into another pig stay and kept in groups in loose housing boxes.

From this time on,, approx ately as of the 10th week, all pigs were exclusively fed with very finely ground fattening feed. In literature the administration of such a feed is indicated besides stress factors as the decisive cause of the formation of mucosal changes, such as hyperkeratoses, erosions or ulcera in the gastric mucous membrane of the pig.

Both groups of animals were administered 3 to 6% whey protein with the drinking water during the entire fattening period till the slaughtering of the pigs after a fattening period of 10 weeks.

The changes of the gastric mucous membrane of pigs in the area of pars proventri cular s was evaluated according to pathological-anatomic criteria.

Table 3 shows the frequency of the evaluation scales (0 to +++) after a 10 weeks feeding of very finely ground feed rations and their percentages (%).

Table 3 0 + ++ +++ 6 6 8 8 control group 21,4% 21,4% 28,6% 28,6% n = 28 10 7 6 2 40% 28% 24% 8% test group n = 25.

In Figure 4 it is graphically represented that in the animals treated with whey protein tw ce as much animals had an intact mucous membrane than in the controls (the two left columns in Figure 4 with the designation "0" which is used for an intact mucous membrane). The two right columns of Figure 4 reveal on the other hand that serious changes of the mucous membrane, e.g. erosions, which are designated with "+++" occurred by 70% less frequently than in the control.

The exact figures of the macroscopic evaluation of the gastric mucous membranes of the exper mental animals as they are revealed by table 3 show that low grade, flat hyperkeratoses (designated with "+") or moderate hyperkeratoses with pointed serrations (designated with "++") occurred with the same frequency in both groups. However, the changes with "+++", i.e. the erosions, are decisive, whose prevention in this test with pigs was possible still more clearly than in the experiments described in examples 1 and 2.

Exj3_mj)L e__ Pej_f_o_rmaji_c_e_ _f__I mmu_n i_s a_t_i_qn__p_f__Ca_t_t.I.e As antigen, a protein is used which is not present in the calf food, especially not in the milk replacer with which the calves are fed after birth. However, the antigen must be present in the high-energy food which is given to the calves in about the eighth to tenth week after birth. The ulcerative lesions preferably develop during the change t high-energy food, usually at an age of four to ten weeks. These disorders are in connection with the mentioned lack of immunization of animals regarding the proteins being present in the high-energy food. The main constituents of the high-energy food for ruminents are mainly corn, wheat soya, barley, grassilage and hay. These constituents may change according to the degree of intensive breeding.

The first intramuscular injection of the used antigenic substance will be given in the first four weeks after the birth of the calf. A second injection will follow in the sixth week and a third injection at the time when high-energy food will be given in the eighth to tenth week. In the first week of breeding the caLves receive milk and starter diet. They will receive high-energy food about the eighth to tenth week. Then, at the time of changing to the high-energy food, the antigenic substance will be added to the food and/or drinking water to use or establish the immunological protection of the gastro ntest nal tract by the above-described immunological mechanism.

The dose of the antigen c substance should be about 0, 1 g test substance per kg bodyweight by ntramuscular application and will be repeated two to three times.

To induce the protective mechanisms, the antigenic substance will then be added to the drink ng water in a dose of up to 1g per 100 ml water.

Performing this procedure, the ulcerative lesions in the gastrointest nal tract of the calves will be prevented or reduced .

According to the above-ment oned handling of the cattle, they will be much better protected against changes of the climate, the food and transports ("Crowding Disease"), especially within the first weeks after birth. The frequently-occurr ng disorders or diseases of the gastrointestinal tract, during these environmental changes, especially the diarrhoea, may well be prevented by the new mechanism. Diarrhoea can be the cause of other bacterial or viral diseases in the gastrointestinal tract.

Concerning the use of antigenic substances in adult ruminants, the required antigenic substances will be applied in bulls and cows during the period of intensive feeding and high lactation. Thus, also in adult animals the more common gastric ulcers, of various stages, may be prevented or reduced.

E x_a_m_p_l e 5 Performance of mmunization of poultry Similar to the immunization of pigs and cattle, domestic poultry, for instance, chickens, turkeys, geese, ducks or turkey-cocks, will be immunized with antigenic substances in the above-mentioned way. The antigenic substances will consist of grain, fish flour, soya or peanuts according to the major sources of food during the later breedi ng.

The parenteral injection of the antigenic substance is performed on the first day after hatching due to practical reasons. This first immunization will be intensified and established by later application of the antigenic substance by putting the antigenic substance into the drinking water in an entral form about one week after the hatching.

Claims (1)

1. 85963/2 27 CLAIMS : A method for the prophylactic and/or therapeutic treatment of disorders and diseases within the gastrointestinal tract of animals, wherein the gastric mucosa is immuniz d by introducing orally or parente.rally an antigenic substance selected from the group comprising plant proteins, animal proteins, human proteins, virus proteins, components of a microorganism, peptides or combinations thereof, and reintroducing orally an effective amount of said antigen. A method according to claim 1, wherein the plant protein is from maize or soya bean. A method according to claim l, wherein the animal protein is whey protein. A method according to claim 3, wherein the whey protein used is 3 to 9% preferably 6% whey protein solution. A method according to claim 1, wherein the human protein is derived from blood or a blood component, such as serum, plasma or hemoglobin. A method to claim 1, wherein the microorganism is a bacterium and wherein the antigenic substance is the bacterium itself, or a component of the cell wall of said bacterium, preferably lipopolysaccharides of the ceil wall of Escherichia coli. 85963/2 28 7. A method according to claim 1, wherein the microorganism is a member of the protozoae. 8. A method according to claim 1, wherein the antigenic substance has been synthesized chemically and is preferably a short peptide, which comprises antigenic determinants for the induction of an immune response. 9. A method according to claim 1, wherein the antigenic substance is a Hapten, preferably nitrophenyl acetic acid (NIP) . 10. A method according to claim 9, wherein the Hapten is coupled to a carrier protein. 11. A method according to any of the aforegoing claims, wherein combinations of 1 or more of the antigenic substances according to any of claims 1 to 10 are used. 12. A method according to any of claims 1 to 11, wherein in addition to the antigenic substance immune modulatory substances are used. 13. A method according to any of the aforegoing claims, wherein the antigenic substance is confectioned for enteral application, preferably as an aerosol, optionally with the addition of pharmaceutically acceptable auxiliary substances. 14. A method according to any of claims 1 to 12, wherein the antigenic substance is confectioned for the parental application, preferably to be used for intramuscular and/or sub-cutaneous administration, optionally with the addition of pharmaceutically acceptable auxiliary substances. 85963/2 29 15. A method according to claim 13 or 14, wherein erosions, peptic ulcers, or cancer in the animal gastrointestinal tract are submitted to prophylactic or therapeutic treatment . 16. A method according to any of claims 11 or 12, wherein diarrhoea, caused by food, which the gastrointestinal tract of the animal has not yet been in contact, is submitted to a prophylactic treatment. 17. Pharmaceutical composition for the prophylaxis and therapy of disorders and diseases within the gastrointestinal tract comprising plant protein, an animal protein, a human protein, a viral protein, a microorganism or a component of a microorganism, or a chemically synthesized peptide, together with a pharmaceutically acceptable carrier. IS. Pharmaceutical composition according to claim 17, wherein the plant protein is a maize or soya bean protei . 19. Pharmaceutical composition according to claim 17, wherein the animal protein is whey protein. 20. Pharmaceutical composition according to claim 19, wherein the concentration of the whey protein is 3 to 9% 21. Pharmaceutical composition according to claim 20, wherein the concentration of the whey protein is approximately 6%. 22. Pharmaceutical composition according to claim 17, wherein the human protein is derived from blood or a blood component. Pharmaceutical composition according to claim 17, wherein the microorganism is a bacteria. Pharmaceutical composition according to claim 17, wherein the component of a microorganism is derived from the cell wall of Escherichia coli. Pharmaceutical composition according to claim 17, wherein the microorganism is a protozoa. Pharmaceutical composition according to claim 17, wherein the antigenic substance has been synthesi?.ed chemically. Pharmaceutical composition according to claim 26, wherein the chemically synthesized protein is a peptide, which comprises antigenic determinants for the induction of an immune respone. Pharmaceutical composition according to claim 1, wherein the Hapten is nitrophenyl acetic acid (NIP) . Pharmaceutical composition according to claim 28, wherein the Hapten is coupled to a carrier protein. Pharmaceutical composition according to claim 17, comprising a combination of the antigenic substances in accordance with any one of claims 17 to 29. Pharmaceutical composition according to any of claims 17 to 30, comprising immune modulatory substances. Pharmaceutical composition according to any of claims 17 to 31 in a form suitable for enteral application. I 85963/2 31 33. Pharmaceutical composition according to claim 32 in the form of an aerosol. 34. Pharmaceutical composition according to claim 33, comprising also pharmaceutical acceptable auxiliary substances . 35. Pharmaceutical composition according to any of claims 17 to 31 in a form suitable for parenteral application. 36. Pharmaceutical composition according to claim 35, in a form suitable for intramuscular and /or subcutaneous injection. 37. Pharmaceutical composition according to claim 36, comprising also pharmaceutically acceptable auxiliaries. 38. Pharmaceutical composition according to any of claims 32 to 37 for the prophylaxis or therapy of erosions, peptic ulcers or cancer in the human or animal gastrointestinal tract . Pharmaceutical composition according to any of claims 17 to 38 for the prophylaxis of diarrhoea caused by food with which the gastrointestinal tract of humans or animals has not yet been in contact. For the Applicants DR. REINHOLD COHN AND PARTNERS By: