AU637045B2 - Preparation with antibody activity and broad spectrum - Google Patents

Description

OPI DATE 24/11/89 APPLN. jID 34171 89 PCT NUiIMBER PflT/nF89/non9 Pcr

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INTERNATIONALE ANMELDUNUj VERK(J11-N1LILHI I1NA(Lh Ur-M Vt-KIKAU I. KUl INTERNATIONALE ZUSAMMENARBEIT AUF DEM GEBIET DES PATENTWESENS (PCT) (51) Internationale Patentkclassifikation 4 (11) Internationale Verorfentlichungsnummer. WO 89/10139 A61K 39/395 C07K 15/06 Al (43) Internationales CO7K 3/02 Veroffentlichungsdatum: 2, November 1989 (02.11.89) (21) Internationales Aktenzeichen: PCT/DE89/00233 (72) Erfinder;und Erfinder/Anmelder (nur fir US) DLCHTELMOLLER, (22) Internationales Anmieldedatumn: 18. April 1989 (18.04.89) Herbert [DE/DE]; Rossertstrasse 14, D-623 1 Sulzbach STEPHAN, Wolfgang [DE/DE]; Philipp-Holzmann-Strasse 84, D-6072 Dreieich LISSNER, Prioritiitsdaten: Reinhard [DE/DE]; Gbnz 18, D-8761 Weilbach (DE).

P 38 13 043.2 19. April 1988 (19,04.88) DE ARNDT, Rildiger [DE/DE]; Caprivistrasse 15, D-2000 88118243.0 2. November 1988 (02.11.88) EP Hamburg- Blanlhenese (DE).

(34)1 Ldnder far die die regionale oder intern anionale Anmeldung eingereicht (74) Anwiilte: BEIL, IHans usw. Beil, Wolff Beil, Adelonworden ist: DE usw. strasse 58, D-6230 Frankfurt am Main 80 (DE).

88121678.2 24. Dezember 1988 (24.12.88) EP (34) Liinderfiir die die regionale oder intern ationale Anmeldung eingereichr (81) Bestimmungsstaaten: AU, BR, DK, F1, HU, JP, KR, NO, worden 1st: DE usw. SU, us.

89104016.4 7. MArz 1989 (07.03.89) EP (34) Liinder.ilr die die regionale oder internationale Anmeldung eingereic/u Verdffentlicht worden 1st: DE usw. Mit internationalem Recherchenbericht, (71) Anmelder (ftir alle Bestimmungssnaten ausser US): BIO- TEST PHARMA GMBH IDE/DE]; Landsteinerstrasse D-6072 Dreieich 674 (54) Title: PREPARATION WITH ANTIBODY ACTIVITY AND BROAD SPECTRUM (54) Bezeichnung: PRXPARAT MIT ANTIKORPERAKTIVITXT UND BREITEM WIRKUNGSSPEKTRUM (57) Abstract A preparation with antibody activity is prepared froma colostrum of non-immunized mammals extracted during the first hours, preferably however during the first 10 hours, following parturition. The colostrum is diluted with water, pasteurized, and after removal of the casein and fat, concentrated and stabilized. The preparation has a high immunoglobulin content (>890 and low anticomplementary activity. It can be administered orally in humans and intravenously in veterinary medicine. It can be used successfully, alone or in combination with other pharmaceutical substances, to treat bacteria- or toxin-induced diseases, in particular severe diarrhea in AIDS patients and other immunological disorders, travellers' diarrhea and toxin-induced infantile diarrhea, gastric and intestinal ulcers, as well as chronic and acute Yersinia infections, and to combat protozoa.

(57) Zusammenfassung Praparat mit Antik6rperaktivittlt, erhaltlich aus Kolostratmilch nicht immunisierter Sliuger der ersten 30, vorzugsweise jedoch der ersten 10 Stunden post partumn durch Verdiinnung der Kolostralmilch mit Wasser, Pasteurisierung, Abtrennung von Casein und Fett, Ankonzentrierung und Stabilisierung, mit einem hohen Immunoglobulingehalt 80 und niedriger antikomplementgrer AktivitAt, zur oralen Applikation beim Menschen und zur intraven6sen Verabreichung im veterindrmedizinischen Bereich. Das Prdparat kann allein oder in Kombination mit anderen Pharmazeutika mit Erfolg zur Behandlung von bakterielloder toxin-bedingten Erkrankungen, insbesondere von schwerer Diarrh5 bei AIDS-Erkrankungen oder anderen immunologischen Defektzustlinden, von Reisediarrh6 und toxin-bedingter Sauglingsdiarrh65, von Magen- und Darmulcera sowie von chronischen und akuten Yersinien-Infektionen, und zur Bekampfung von Protozoen eingesetzt werden.

(11) AU-B-34171/89 -2- 637045 9. A method of preparing immunoglobulin from bovine colostrum from nonimmunised mammals of the first 30, preferably 10 hours post-partum, comprising diluting the colostral milk with distilled water and pasteurising the resulting solution, separating tne fat and breaking down and removing the casein, spray drying the thus obtained colostral whey to produce an immunoglobulin preparation, the entry temperature of the colostral whey to said spray drying being from 145 180 0 C and the exit temperature from said spray drying being from 65 70 0

C.

Dietest Prhrma Gmb.!- Landste*:fstfasse 6072 Drcicich Product with antibody activity and a broad spectrum of action The invention relates to an immunoglobulin product which has high antibody activity and which can be obtained by known processes from colostral milk from nonimmunized mammals. The invention furthermore relates to an agent containing this product or consisting thereof, and to the use thereof for the treatment of bacterial- or toxin-related diseases, especially of diar-oea associated with AIDS or other immunological deficiency states, of travellers' diarrhoea and toxin-related diarrhoea in babies, of gastric and intestinal ulcers and of Z and acute Yersinia infections, and for combating protozoa.

Immunoglobulins a group of highly active and, at the same time, sensitive proteins are used, inter alia, for the prophylaxis and treatment of bacterial or viral diseases and, where appropriate in the form of special compositions, against certain toxins. They can be obtained by relatively elaborate processes from human plasma or, in a somewhat more straightforward way, from milk or colostral milk of mammals.

The group of immunoglobulins obtainable from human plasma includes, for example, immunoglobulin G, a protein which is effective for the prophylaxis and treatment of a wide variety of infections.

The known immunoglobulin-containing products obtained from milk or colostral milk have a somewhat lower purity than those obtainable from human plasma.

They are used predominantly in veterinary medicine for the._ treatment of infectious diseases.

Numerous processes for obtaining immunoglobulins from milk or colostral milk have been disclosed to date.

*For example, US Patent 4,526,715 describes a process for producing immunoglobulins from milk via a 2variety of chromatographic separation steps.

US Patent 3,128,230 relates to a process for the preparation of a product, which can be obtained from milk, for the treatment of various bacterial infections.

US Patent 4,265,925 and US Patent 1,573,995 describe processes for the preparation of protein concentrates from milk serum, there being very extensive denaturation of these proteins because of a heat treatment.

Finally, German Offenlegungs chrift 2,813,284 and USf-O57235 tnd r- boo 7533 EP-A 0,046,909 and EP-A 0,064,103Adescribe colostral milk preparation processes resulting in immunoglobulin-contaiing products as antiinflammatory agents.

The preparation of pharmacologically active immunoglobulin-containing products via the abovementioned processes starts from milk or colostral milk from hyperimmunized mammals, especially cattle. This therefore entails, in order to achieve antibacterial or antiviral efficacy against particular pathogens, initially injecting into the animals one or more strains of bacteria, or the antigens thereof, which induce an antibody or a group of specific antibodies.

The activity of the products obtained in this way corresponds to the type and number of strains of bacteria administered.

To prepare a product which is obtainable from milk and has a specific antibody activity (for example against bacterial antigens) the mammal is also initially immunized with specific antigens. The milk which is then obtained has a high titre of antibodies against the specific antigen, with the milk acting as carrier medium.

The invention has the object of preparing an immunoglobulin product which has high purity and a high antibody activity by a straightforward and economic process which can be used in the area of human medicine as well as in that of veterinary medicine without it being necessary previously to carry out hyperimmunization in order to prepare specific antibody titres.

This object is achieved according to the invention by an immunoglobulin preparation prepared from bovine colostrum of the first 30, preferably 10, hours post-partum, in which the colostral milk is diluted with distilled water and then pasteurised, the fat is separated and the casein is broken down and removed, said preparation being obtained by spray drying of the thus obtained colostral whey, the entry temperature of the colostral whey to said spray drying being from 145 180°C and the exit temperature from said spray drying being from 65 700C.

This results in an immunoglobulin-containing product which has high purity and an immunoglobulin content of >80% based on the total protein content, as well as very low proportions of fat and lactose and which has a higher antibody activity than the human immunoglobulin product "Pentaglobin"(R) hitherto known as the most effective. By reason of the extremely low anticomplementary activity, this product according to the invention is suitable for oral administration to humans and for intravenous administration in veterinary medicine.

The product, obtained, for example, after purification and separation processes as are customarily used in milk processing can be made available in a form which is stable on storage, either as a solution or as a powder.

For this purpose, colostral milk, for example from cattle in the first hours post partum, is diluted with distilled water in the ratio 1:3. After pasteurization, the fat is removed from this solution and subsequently casein is precipitated and removed, for example by filtration. The whey obtained in this way is concentrated to a desired content. A sterile filtration can be carried out to stabilize the product, resulting in a sterile solution of the product.

Stabilization by means of spray-drying results in the product in the form of la powder. It has emerged from this, surprisingly, that no denaturation of the proteins takes place at the temperatures required for the spray-drying (entry temperature about 145°C to 1800C, exit temperature about 65-70°C). Thus, under these conditions of spray-drying, there is no extreme aggregate formation and thus no increase in the anticomplementary activity of the proteins as was actually known and was to be expected at such temperatures. Thus, even when TFL,. this stabilization method is used, the product obtained has an unaltered high 4 q -4immunoglobulin content of >80% with an unaltered low anticomplementary activity and complete biological activity.

In addition, it is possible before the spray drying step has been carried out, to carry out a precipitation reaction with octanoic acid. This achieves a further enhancement of the immunoglobulin content to more than 90% without having an adverse effect on the anticomplementary activity.

An immunoglobulin-containing product with high antibody activity can also be prepared by other, likewise known, processes from colostral milk from the first 10 hours post partum. It is possible to use for this, for example, a i process as specified in German Patent 3,432,718. For this purpose, colostral milk from non-immunized cattle is initially acidified and subjected to coarse filtration (removal of casein), and subsequently diluted with sodium chloride solution. The suspension obtained in this way is subjected to tangential filtration, followed by concentration and then neutralization. To stabilize the concentrated whey, it is thus possible to carry out a spray-drying, and, where appropriate, an octanoic acid precipitation to increase the purity of the product.

Preparation of the immunoglobulin product Example 1 110 kg of deep-frozen bovine colostral milk from the first 10 hours after calving were thawed at 5-10oC and dilutcd with distilled water in a ratio 1:3. The milk was then pasteurized, and subseqi ntly the fat was removed This resulted in 270 kg of skimmed;milk. The latter was acidified with 1N HCI, and the precipitated casein was removed through filter cloths. The supernatant (186 kg) obtained in this way underwent ultrafiltration, resulting in 38 kg of concentrate.

This concentrate was spray-dried for the stabilization. The spray-drying is Scarried out at an entry temperature of 175°C and an exit temperature of 650C.

This resulted in a dry powder which contains immunoglobulin in 83% purity, corresponding to a total yield of 3.8 kg of immunoglobulin.

The antibody activity of this product before spray-drying corresponded to that after spray-drying.

Example 2 13 of bovine colostral milk which had been taken in the first 5 hours after 0* 0 1 u ii.

I-.

calving and had been deep-frozen were thawed. The pH was 6.27. This was adjusted to 4.8 by acidification with 700ml of 1 N HCI. The suspension obtained in this way was heated at 40°C for 30 minutes and then stored at 4°C overnight.

Subsequently, to remove coarse particles, the suspension was pumped through a gauze filter made of nylon gauze into a storage vessel and there diluted to a total volume of 26 I with 0.9% strength sodium chloride solution.

The suspension obtained in this way was then subjected to the first tangential filtration through a hollow-fibre cartridge with a mean pore size of 0.4 prm, a surface area of 3 m 2 and a fibre diameter of 1.2 mm by diafiltration using 100 I of a 0.9% strength sodium chloride solution under a pressure of 0.2 to 0.6 bar above atmospheric.

Even while the first tangential filtration was in progress, the resulting diafiltrate was subjected to a second tangential filtration using a system consisting of three hollow-fibre cartridges each with a separation limit of 10,000 D and a surface area of 1.4 m2. In this system the diafiltrate from the first tangential filtration was initially concentrated and then underwent diafiltration using 100 i or a 0.9% strength sodium chloride solution and was finally concentrated to a total volume of 25 I. The filtrate containing the low molecular eight components was removed.

Example 3 200 ml of an immunoglobulin solution prepared as in Example 1 were, before the stabilization step, mixed with octanoic acid at pH 4.8 and a temperature of 230C for 5 hours. The precipitate was subsequently removed by filtration, and the supernatant was dialyzed against 0.9% strength NaCI solution, and spray-dried.

i The immunoglobulin content in the product obtained in this way was The anticomplementary activity corresponded to the low value of a human IgG preparation tolerated on intrevenous administration.

Example 4 200 ml of solution with a protein content of 5% were prepared from a (spray-dried) immunoglobulin powder prepared as in Example 1. A solution of Sthis type (5 g of powder stirred into H 2 0 ad 100 ml) has the following I 1 1 -6composition: Total protein including lgG IgA 1gM 4.2 g/1l00 ml 3.0 g/1 00 ml 0.35 g/100 ml 0.96 g/100 ml 4.6 1.0% CAF' pH lactalbumnin This solution was incubated with 2.5% octanoic acid at pH 4.8 and T= 23 0 C for 5 hours. The precipitate was removed by centrifugation, and the supernatant was subjected to diafiltration. The anticomplementary activity corresponded to that of a human lgG product of the 0**e 7 same concentration for intravenous administration.

The immunoglobulin product obtainable by the described or analogous processes has a higher antitoxin activity than Pentaglobin.

Neutralization of bacterial toxins The neutralization of haemolyzing bacterial toxins was determined in the haemolysis inhibition test (HIT) using toxin-containing supernatants of various bacteria.

Example An overnight culture in brain heart infusion nutrient broth of Staphylococcus aureus Smith 3 was centrifuged at 10,000 x g for 10 min, and the supernatant was subjected to sterile filtration. The sterile supernatant, which contained haemolyzing toxins, from this culture was used for the haemolysis inhibition tests.

A 5% strength solution of an immunoglobulin product prepared as in Example 2 was tested for the toxin-neutralizing effect. For this purpose, 0.1 ml of washed human erythrocytes was incubated with 10 pl of Staphylococcus aureus toxin in 900 to 990 pl of NaCl at 37 0 C for 30 minutes, in each case 10-100 4l of immunoglobulin solution being added. The comparison preparation was a 5% commercially available immunoglobulin preparation (Pentaglobin). Figure 1 shows the result, where the measure used for the haemolysis or the inhibition thereof was the measured extinction at 542 nm.

Example The procedure was that stated in Example 6 but in this case the toxin-containing supernatant from a Gramnegative organism Ps. aeruginosa was used to test for toxin neutralization. The haemolysis was measured as in Examples 5 and 7 in the range between 0 and 100 l of the immunoglobulin solution diluted 1:10 in NaCl Example 7 The toxin-containing supernatant of Staphylococcus aureus was used for the investigation described hereinafter. The toxin neutralization was tested using a product strength solution) prepared as in Example 1 I r S 8 -8- (spray-dried). Pentaglobin in the same concentration was used for comparison in the HIT. The haemolysis was measured in the range from 0 to 100 1p of immunoglobulin solution diluted 1:10 in NaC1).

The results of the experiments of Examples 5-7 are depicted in Figures 1-3.

Figure 1 shows the neutralization of bacterial haemolyzing of toxins of Staphylococcus aureus Smith 3 as in Example 5 by immunoglobulin from bovine colostrum prepared as in Example 2 and by Pentaglobin as comparison.

Figure 2 shows the neutralization of bacterial toxins of Pseudomonas aeruginosa as in Example 6 by immunoglobulin from bovine colostrum prepared as in Example 2.

Figure 3 shows the neutralization of bacterial toxins of Staphylococcus aureus as in Example 7 by bovine colostrum immunoglobulin prepared as in Example 1 and by Pentaglobin as comparison.

It is evident from Figures 1-3 that the immunoglobulin preparation obtained from bovine colostrum brings about a more effective inhibition of the toxininduced haemolysis than does Pentaglobin, which is obtained from human plasma and is of comparable composition.

Table I summarizes the results of the experiments of Examples 5-7 9 Tale I Example Concentration Concentration at haemolysis haemolysis -PG (comparison)* 50 l 93 pl RG (invention)** 10 1l 50 l 6 RG (invention) 18 1 35 1 7 PG (comparison) 42 l 80 /i 1P (invention) 10 Al 40 1l PG Pentaglobin RG immunoglobulin from bovine colostrum Examples 5 and 7 demonstrate that for 50% haemolysis (toxin neutralization) the amount of the known pentaglobin which is required is three times that for the product according to the invention. Twice the amount of Pentaglobin must be used to achieve 80% toxin neutralization.

It is evident from Example 6 that the immunoglobulin according to the invention brings about even at very low concentrations (40 Al) almost 100% toxin neutralization, even of the toxins of Gram-negative bacteria.

100% haemolysis is not reached with Pentaglobin in the concentration range under consideration (maximum 100 Al of a 5% strength solution). Since the immunoglobulin product prepared from colostral milk is, in contrast to Pentaglobin, not 100% pure the therapeutic index derived from the abovementioned results for the antitoxin according to the invention is at least twice that of the known Pentaglobin.

As is also evident from Table I and Figure 3, the process chosen for preparing the product has no effect on the activity thereof. It can be obtained both by known methods of the dairy industry and by more specialized 1 preparation processes (for example tangential filtration).

I L 10 The product according to the invention can be administered intravenously in the arsa of veterinary medicine and can be administered orally to humans. It has no anomalous toxicity, as is demonstrated in Examples 8 to (0 which follow, and can be stored both liquid and in the solid form.

Tolerability anomalous toxicity Example Groups of 5 mice received a 5% strength immunoglobulin solution tested as above (as in Examples 5-7) administered intraperitoneally (0.1 ml/animal). The animals were observed for 3 hours and kept for 5 days after the administration.

Result: No animal died, no immediate reaction, no reaction up to 3 hours after administration, no changes.

Dosage: 0.29 g/kg (weight of mouse: 17 g).

Example Groups of- 5 mice received 0.5 ml of the solution, pH 4.6, administered orally by gavage and were observed for 3 hours after administration and were kept for 5 days after the administration.

Result: No animal died, no immediate reaction, no reaction up to 5 hours, no changes.

Dosage: 1.47 g/kg; (weight of mouse 17 g) Example Groups of 3 mice received 1.0 ml of a 10% susp&et sion, pH 4.6, administered orally by gavage.

Result: No signs were observed immediately or up to 1 hour, or after 3 and 24 hours. All the animals were unchanged after 5 days.

Dosage: The dose of 1.0 ml of 10% solution (0.1 g/animal) corresponds to a dosage of g/kg or 350 g/70 kg (weight of mouse 20 g).

The immunoglobulin product according to the invention which can be obtained from colostral milk can be prepared in a very straightforward and economic manner and has an unexpectedly high antitoxin activity against 11 a wide variety of bacterial toxins. It is distinguished by excellent tolerability (no anomal toxicity) and can be stored in high purity both in liquid form and as a powder in stabilized form.

Example (I It has been found, surprisingly, that the products according to the invention are excellently suited for the treatment of severe diarrhoea as sequelae of HIV infections or of other immunological deficiency states.

On administration of the products according to the invention there was, within a very short time, a considerable reduction in the number of attacks of diarrhoea each day, and even complete cessation of diarrhoea, as is shown in Table II which follows. 10 g of the product according to the invention was administered each day for a period of 10 days to each of the patients suffering from AIDS.

Table II AL.acks of diarrhoea Body weight kg per day Patient Before Days after start of Before After therapy therapy 1 3 5 10 therapy 10 5 1 1 1 42.0 n.t.

6 1 0* 0* 0* 84.4 85.5 10 5 2 2 1 62.3 62.8 normal stool The products according to the invention can be administered alone or in combination with other antidiarrhoeals or other agents necessary or suitable for the therapy, and result within a very short time in a significant reduction in the frequency of diarrhoea and thus in a considerable improvement in the general condition of the patient.

Example The products according to the invention are, surprisingly, also suitable for the treatment both of 12 so-called travellers' diarrhoea and of toxin-related diarrhoea in babies.

It has been found in laboratory investigations that the product according to the invention contains antibodies against, inter alia, the 40 KD protein of verotoxin 1 (VTI). VT 1 or shiga-like toxin 1 is, besides verotoxin 2, the most important cytotoxic exotoxin of E.

coli pathogenic for babies (EPEC) and of enterohaemoprhagic E. coli (EHEC).

Furthermore, the product according to the invention contains antibodies against 30 KD and 20 KD proteins of the heat-labile enterotoxin of enterotoxin-forming E.

coli (ETEC), one cause of-travellers' diarrhoea.

Hence the product according to the invention contains antibodies against virtually the entire spectrum of toxins of enteropathogenic coli organisms (ETEC, enterotoxinogenic; EPEC, pathogenic for babies; EHEC, enterohaemorrhagic). This is a considerable advantage for the particular reason that these diseases are common and may, at least in part, be the consequence of an antibiotic therapy, or the antibiotic administration often brings about adverse effects in the patient owing to toxin release.

The results of the investigations carried out using the immunoblotting technique at various concentrations are depicted in Figure 4.

Example (3 Duodenal and gastric ulcerations'associated with gastritis have hitherto been treated with bismuth salts or antibiotics. The straightforward therapeutic methods which have few side effects (bismuth salts) are of only low efficacy, whereas the effective antibiotic combination regimens are reliable but scarcely practicable for routine use because of the side effects. Accordingly, there is a need for straightforward therapeutics which have few side effects and are, at the same time, effective. According to current knowledge, Campylobacter pylori is of pathogenic importance for these disorders as S well as for non-ulcerative dyspepsia. Characteristic of 13 C. pylori are an outer membrane protein of 120 KD, a specific flagellar antigen (flagellin) of 69 KD and the urease of 45 KD.

It has now been found, surprisingly, that the product according to the invention also possesses antibodies in high concentrations against these three proteins.

The immunoblotting technique was used for testing. The product according to the invention with a protein content of 4 mg/ml was tested undiluted and in dilutions of 1:10 and 1:100. Detection was effected by bovine IgG conjugated by alkaline phosphatases. As is evident from the blot depicted in Figure 5, the product according to the invention reacts at a dilution of 1:10 with all three abovementioned proteins and in a dilution of 1:100 still clearly with the flagellin and the urease.

Furthermore, the product according to the invention also contains a large number of antibodies which react with the C. pylori preparation.

Hence, surprisingly, the product according to the invention is also excellently suited for the treatment of infections with Campylobacter pylori.

Example 14- Yersinia infections result not only in an acute enteric course but also, in particular, in subacute and chronic disease forms which, as a rule, are associated with a negative culture finding, but with pathogens still being detectable by immunohistology. The most important disorders with a positive IgA antibody test indicating existing infection are reactive arthritis, erythema nodosum, uveitis, chronic enteritis and the like.

Experience has shown that Yersinias contain, irrespective of the species, so-called YOP antigens (inter alia: YOP 2b with 47 KD, YOP 3 with 37 KD, YOP with 26 KD). Using these isolated antigens, the product according to the invention has been tested in a Western blot. The product was used for this undiluted (protein content 4 mg/ml) and in dilutions of 1:10 and 1:100. As is evident from Figure 6, the product reacted with all 14 three antigens at all the concentrations used. The staining in these tests was carried out with a specific anti-bovine IgG (H and L chains) conjugated to alkaline phosphatase, in a dilution of 1:1500 and 1:3000.

Because the antigens detected by this are independent of the serotype and species of the Yersinia, the product according to the invention is also excellently suitable for the treatment of Yersinia infections.

Example The products, according to this invention are, surprisingly, also suited for combating protozoa, as for example Cryptosporidia, Isospora belli or ToxaplasmaGondii against which not in all cases of such diseases remedies were known up to now.

Cryptosporidia lead to serious diarrhoea in patients with immune deficiencies, which could not effectively be treated with known medicines. Four patients with heavy diarrhoea, in whose excrements Cryptosporidia could be identified, were give the product according to this invention in the form of a drinkable solution for 10 subsequent days in a dose of 10 g per day. After finishing thistherapy Cryptosporidia specific antigens could not be shown with any of these patients.

Claims (10)

1. Immunoglobulin preparation from bovine colostrum from non-immunised mammals of the first 30, preferably 10 hours post-partum, in which the colostral milk is diluted with distilled water and then pasteurised, the fat is separated and the casein is broken r down and removed, said preparation being obtained by spray drying of the thus obtained colostral whey, the entry temperature of the colostral whey to said spray drying being from 145 180°C and the exit temperature from said spray drying being from 65 700C.

2. A preparation as claimed in claim 1 or 2, wherein the colostral whey is concentrated prior to spray drying.

3. A preparation as claimed in claim 1 or 2, wherein prior to said spray drying the colostral whey undergoes octanoic acid treatment.

4. A preparation is claimed in any one of claims 1 to 3, wherein said entry temperature is about 145 0 C and said exit temperature is about 650C. A method of treating an ailment selected from diarrhoea in acquired immune deficiency syndrome or in other immune deficiency conditions, travel diarrhoea, toxin-conditional infant diarrhoea, stomach ulcers, intestinal ulcers, non-ulcerous dyspepsia, chronic yersinien infections and acute yersinien infections, comprising administering to a patient in need of such treatment an effective amount of the preparation of any one of claims 1 to 4.

6. A method of treating an ailment selected from diarrhoea in acquired immune deficiency syndrome or in other immune deficiency conditions, travel diarrhoea, toxin-conditional infant diarrhoea, stomach ulcers, intestinal ulcers, non-ulcerous dyspepsia, chronic yersinien infections and acute yersinien infections, comprising administering to a patient in need of such treatment an o: effective amount of a compound comprising the preparation as claimed in any one of claims 1 to 4 and a pharmacologically acceptable carrier or excipient.

7. A method of combating protozoa comprising administering to a patient in need of such treatment a preparation as claimed in any one of claims 1 to 4.

8. A method of combating protozoa comprising administering to a patient in need of such treatment a compound comprising the preparation as claimed in 16 any one of claims 1 to 4, and a pharmacologically acceptable carrier or excipie t.

9. A method of preparing immunoglobulin from bovine colostrum from non- immunisec mammals of the first 30, preferably 10 hours post-partum, comprising diluting, the colostral milk with distilled water and pasteurising the resulting solution, separating the fat and breaking down and removing the casein, spray drying the thus obtained colostral whey to produce an immunoglobulin preparation, the entry temperature of the colostral whey to said spray drying being from 145 180°C and the exit temperature from said spray drying being from 65 700C. A method as claimed in claim 9, wherein prior to said spray drying the colostral whey is concentrated.

11. A method as claimed in claim 9 or 10, wherein prior to said spray drying the colostral whey undergoes octanoic acid treatment.

12. A method as claimed in any one of claims 9 to 11, wherein said entry temperature is about 145 0 C and said exit temperature is about 650C. Dated this 1st day of February, 1993. BIOTEST PHARMA GMBH WATERMARK PATENT TRADEMARK ATTORNEYS LEVEL 2, THE ATRIUM, 290 BURWOOD ROAD, HAWTHORN, VICTORIA 3122. a a a AU3417189.WPC DOC024 •r O U7- Abstract Product with antibody activity and a broad spectrum of action, Product having aptibody activity and obtainable from colostral milk of non-immunized mammals in the first but preferably first 10, hours post partum by dilu- tion of the colostral milk with water, pasteurization, removal of casein and fat, concentration and stabiliz- ation, and having a high immunoglobulin content and low anticomplementary activity, for oral adminis- tration to humans and for intravenous administration in the area of veterinary medicine. The product can be used alone or in combination with other pharmaceuticals for the successful treatment of bacterial- or toxin-related diseases, especially of severe diarrhoea associated with AIDS or other immunological deficiency states, of travel- lers' diarrhoea and toxin-related diarrhoea in babies, of gastric and intestinal ulcers, as well as of chronic and acute Yersinia infectionsand for combating protozoa.