WO2003030918A1

WO2003030918A1 - Pharmaceutical product or food supplement and intermediate product to be used therewith

Info

Publication number: WO2003030918A1
Application number: PCT/NL2002/000640
Authority: WO
Inventors: Jeroen Johannes Maria Van Den Berg; Robert Johan Joseph Hageman; Eric Alexander Franciscus Van Tol; Adrianus Johannes Maria Vriesema
Original assignee: Nutricia NV
Current assignee: Nutricia NV
Priority date: 2001-10-08
Filing date: 2002-10-07
Publication date: 2003-04-17
Anticipated expiration: 2004-04-08
Also published as: NL1019143C2

Abstract

The present invention relates to a pharmaceutical product or food supplement which contains a preparation from blood plasma as the active component, in which the preparation contains a larger amount of complement system (CS) and/or antibodies (immunoglobulins) than is normally present. The present invention furthermore relates to the pharmaceutical product in a form which is suitable for medical application and to the food supplement in solid or liquid form, as well as to the intermediate product to be used.

Description

Pharmaceutical product or food supplement and intermediate product to be used therewith.

The invention relates to a pharmaceutical product or food supplement which contains a preparation from blood plasma as the active component. The invention further relates to the pharmaceutical product in a form which is suitable for medical use and to the food supplement in solid or liquid form, as well as to an intermediate product to be used.

Food supplements containing substances from animal plasma as the active component are known from US patent No 6,004,576. Said food supplements are used for effecting a weight increase in animals and for stimulating their growth. By processing the blood plasma in a different manner it becomes possible to obtain plasma fractions which can be used as the active component in a pharmaceutical product. Thus, the invention in particular relates to a pharmaceutical product and food supplement containing a specific fraction which can be separated from animal blood. The preparation thus obtained can be administered enterally or topically; in particular, it can be successfully administered to patients who suffer from diarrhoea, from various wounds or infections or from a specific deficiency of the immune system.

According to the invention, the pharmaceutical product or food supplement comprises a preparation which contains a larger amount of complement system (CS) and/or antibodies (im unoglobulins) than is normally present as the active component. The preparation in particular contains a larger amount of complement system (CS) than is normally present. In practice the phrase "larger amount of complement system than is normally present" is understood to mean an amount such as the amount present in the blood preparations that are (commercially) known so far. The blood plasma of particular animals contains a particular amount of complement system or antibodies. This amount depends in the first place on the animal species in question, for example pigs, cattle, horses or poultry. Furthermore, the amount will vary within a specific species. Because the blood plasma is obtained from several animals, however, and the blood plasma from said animals is combined, the variation within the specific species will be eliminated for the larger part. The normal way of processing such blood operations results in the loss of a large part of the action of the immunoglobulins, and in particular of the components of the complement system. According to the invention, the blood plasma thus obtained contains a larger amount of complement system (CS) and/or antibodies (immunoglobulins) than is normally present. It will be understood that the larger the amount of complement system (CS) that is present in addition to the amount that is normally present, the greater the effect will be. It is in particular important that a larger amount of complement system (CS) be present in the preparation. It is preferable to obtain the complement system (CS) or the specific antibodies in an amount larger than the amount that is normally present by immunising animals and further processing the blood plasma from the immunised animals.

According to the invention it is important, therefore, that said blood plasma contains a higher amount of antibodies (immunoglobulins) than normally present, as well as a number of proteins which together form the complement system (CS) .

According to the invention it is furthermore important that the preparation contains a complement system and/or antibodies which have not been denatured. Preferably, an intermediate product from blood plasma containing more than 2.6 g of CS, in particular more than 3.25 g of CS, or more than 4 g of CS per 100 g of (intermediate) product forms the basis for the preparation of the pharmaceutical product or food supplement. The immunoglobulins may be of the so-called 6-type

(dominant in blood from cows and pigs) or of the Y-type (dominant in blood from poultry) .

The immunoglobulins may be specific against toxins such as the anti-LT-IgG (heat-unstable toxin) and immunoglobulins which are specific with regard to cholera toxin or the toxins that are developed from or by E-coli or other pathogens. Said antibodies with regard to toxins can be found in normal blood, viz. blood from non- hyperimmune animals.

The immunoglobulins may also be specific against a larger number of pathogens, such as Helicobacter pylori, Pseudomonas aeruginosis, staphylococcus aureus, E. coli, Salmonella and other Gram- negative or Gram-positive micro-organisms, as well as fungi such as Candida. Titres in the blood of normal (non- hyperimmunised) animals are too low to be active after being administered to humans, so that the animal must be subjected to a hyperimmunisation treatment, using methods which are known per se in this field. It is preferred to use the method that will be explained in more detail hereinafter, in particular if the animal is subjected to a hyperimmunisation treatment with a cocktail of relevant micro-organisms.

Preferably, the active component of the blood also contains proteins, indicated as the "complement system", or the relevant components thereof. The complement system (CS) is a group of glycoproteins which are capable of interaction under specific conditions, which will lead to an enhanced immune response. CS is in particular involved in the activation of immune cells, such as neutrophilic granulocytes and monocytes. CS occurs in the blood of many animals, although the type and the concentration of the various components of CS vary in dependence on the specific species. CS contains proteins which are involved in the activation of the C3-protein, such as the immune complexes Cl, C4 and C2 and the factor B, P and D, which comprise products of bacterial origin. CS also comprises enhanced proteins involved in cytolysis, such as C5-C9 proteins and bioactive fragments or complexes thereof, in particular C5a and C5i67. CS comprises proteins which are involved in the regulation (including the inhibition) of CS, such as Cl-inhibitors, C4 binding protein factor H, I, CD59 (protectin), anafylatoxin-inactivator and S-protein. Under non-inflammatory conditions, the system is in the non-activated form.

For the application according to the invention it is important that CS can act in the desired manner when administered to the patient. The essential components must be present in sufficient amounts, which can be determined by the doctor in attendance by carrying out suitable measurements, for example by carrying out a complement-lysis determination. A high degree of synergy can be measured. Essential substances are C3-protein, which makes up more than 30 wt.% of the complement system, and the C5-C9 proteins (C5, C6, C7, C8, C9), which together make up more than five wt.% of CS. The weight ratio of the components of the complement system to the amount of globulins in the product must range between 1:5 and 1:20, in particular between 1:7 and 1:16. The ratio of the amount of components of the complement system without inhibitors of CS to the amount of immunoglobulins ranges between 1:8 and 1:25, in particular between 1:10 and 1:20.

It has become apparent that a very effective complement system can be obtained in particular from the blood of hyperimmunised animals.

Preferably, the blood component also contains growth factors, such as insulin-like growth factor (IGF), transforming growth factor (TGF), in particular TGF- beta combined with "Latency Associated Protein (LAP)", platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and keratinocyte growth factor (KGF) .

The blood product preferably does not contain any fibrinogen (serum) or other components that interfere with the preparation of liquid products from the compotent. Furthermore, the blood product should not contain any impurities such as micro-organisms (pathogens, viruses) or prions.

The blood product or the preparation is prepared by carrying out methods of such a nature that the active components are not affected. Both immunoglobulins and in particular the complement system are heat sensitive. Consequently, conventional treatments, which may be used for the isolation and in particular for the concentration of proteins, lead to significant immunoglobulin losses, viz. 20-50% and even more for the components of the complement system. For use in foodstuffs for human beings it is desirable to carr out an additional purification and concentration step by removing, for example, albumins. In the preparation of a suitable means for human nutrition it is strongly preferred to avoid heat treatments that have been conventionally used so far, such as pasteurisation, vaporisation or sterilisation. Within this framework, alternative treatments must be used, such as chromatography, electroforesis, centrifugation or filtration techniques, or a combination thereof. It is preferred to use a dialysis treatment, in particular when combined with differences in the voltage potential across the membranes and with the use of the so-called counterflow, in which the direction of flow of the starting material on one side of the membrane is opposed to direction of flow of the liquid flowing to the other side of the membrane. It is in particular preferred to dry the component before it is further used in the preparation of the food supplement or the pharmaceutical product. The drying of the liquid blood product can also take place directly after the filtration step, but it is preferably carried out on the concentrated material (in which said concentration takes place through dialysis), the standardisation step can take place after said dialysis with regard to the adjustment of the pH value and the adjustment of the salt concentration for the stabilisation and standardisation of the product. Suitable drying techniques are freeze-drying or spray-drying, using specific drying conditions, or a combination thereof. A suitable spray-drying technique is described in Dutch patent No 1016981.

The organoleptic properties of the final product are appreciated by the consumer or the patient. Depending on the manner of preparation, specific amounts of the components are present in the product according to the invention. After centrifugation and removal of fib inogen, liquid plasma remains, with more than 60%, in particular more than 70%, especially about 80% of the dry material being protein. Only about 20% are immunoglobulins. The removal of albumins and the use of additional steps as mentioned above results in a product in which more than 70 g, in particular more than 80 g/100 g of dry material is a protein-like material, of which 55-93 g, in particular 65-90 g/100 g of proteins are immunoglobulins. The remainder consists of the components from the complement system, growth factors and other proteins. When blood from cows or pigs is used, the immunoglobulins will be of the G-type (IgG). Part thereof, in particular 0.01-5%, are antibodies against preselected toxins or pathogens.

The concentration of the components in the final product can be computed by a person skilled in this field by assuming that about 0.001-0.002 g of plasma-protein according to the invention per kilogram of body weight, in particular 0.003-0.014 g/kg of body weight, must be consumed per day in order to be effective. Said amounts are relatively low, because the complement factor from the blood plasma and the immunoglobulins against pathogens, as present in blood plasma, appeared to have a synergetic effect.

In the situation that the product is a preparation to be locally administered, the concentration of the protein must be about 0.1- 10 g, in particular 0.2-8 g, of plasma proteins per litre.

The active component according to the invention is important for the preparation of products to be used as food supplements or as a pharmaceutical product for use with humans and animals, which animals include in particular pigs, cows, poultry, horses and pets such as dogs and cats.

The active component exhibits an activity which is favourable in connection with the healing of wounds, the recovery and maintenance of tissue, in particular epithel tissue in the gastrointestinal system, the respiratory system or the skin, the normalisation of the intestinal flora and the prevention or treatment of infections that may occur with wounds and in the gastrointestinal and respiratory systems. Infections in the gastrointestinal system may occur in the intestines, for example in the case of infection-triggered diarrhoea, in the stomach in the case of stomach ulcers and in the throat. An example of an infection in the respiratory system is RSV-infection or Pneumonia. These infections can manifest themselves when the immune function is compromised, for example during the final stages of AIDS, after surgery with elderly people, in the case of prolonged sickness, but also during or after cancer therapy (chemotherapy or radiotherapy).

The active component is also capable of influencing the flora in the gastrointestinal system. The growth of specific icro- organisms is impeded, enabling the flora in the intestines to normalise, for example in the case of diarrhoea or bacterial overgrowth.

The integrity of the tissue in the gastrointestinal system is in particular important in order to obtain a sufficiently great barrier function.. During and after surgery and during illnesses in which the immune function has been affected it is of major importance to prevent infections and sepsis.

The improvement of the healing process of wounds constitutes another aspect of the invention. Said wounds may occur on the skin, for example after traumatic experiences such as fire, cuts or infections and in the case of eczema, but also on deeper surfaces, for example after decubitus, surgery or radiotherapy. The pharmaceutical preparation may also be used to prevent infections as referred to above.

The application determines the form of the product according to the invention. The treatment of the skin is preferably carried out by local application. This can take place by using a gel, an emulsion, an ointment or other forms known to those skilled in this field.

The treatment of the respiratory system can take place by administering the pharmaceutical product in the form of a spray or an aerosol, for example. The treatment of the gastrointestinal system can take place by enteral administration of the products. Formulations in liquid form as well as formulations in dry form are suitable. Examples of relatively dry formulations are tablets, capsules, suppositories and the like. In addition to that, the products may have a low dry matter content, for example lower than 50% and in particular lower than 20%. Emulsions, gels and mixtures are the most suitable forms in that case.

The invention will be explained in more detail hereinafter by means of the following examples.

Example 1 The blood of pigs was hyperimmunised by means of an extract from the faeces of children who suffered from diarrhoea. The blood was further treated by removing fibrin and albumin therefrom, followed by filtration and dialysis. Dry matter (100 g) consisted of 93 g of proteins and 7 g of remaining substances such as lipids, with 86 g/100 g of protein consisting of immunoglobulins and 9 g/100 g of proteins consisting of components belonging to the complement system.

Example 2

A liquid intended for being drip-fed to patients in a hospital was prepared as follows. A composition of 100 ml having an energy value of 420 kJoule contained the following components:

Protein 4.0 g, viz. 3.9 g of milk proteins (80% casein, 20% whey) and 0.1 g of protein fraction from Example 1.

Carbohydrates 12.3 g consisting of 11.1 g of polysaccharides from maltodextrins, 0.7 g of maltose and 0.5 g of remaining substances. Lipiden 3.9 g, consisting of 0.28 g of saturated fatty acids, 2.2 g of monosaturated fatty acids, 1.1 g of polyon-saturated fatty acids, the remainder being glycerol as tπ^'glyceride.

Minerals, spore elements and vitamins in amounts which are comparable to the recommended amounts in known preparations that are generally accepted for enteral clinical feeding.

Example 3

After enrichment in water, a powder obtained by spray- drying according to Dutch patent No 1016981 yielded the liquid formulation as indicated in Example 2. The activity of the complement system amounted to more than 75% of that prior to the drying step.

Example 4

A formulation intended for being administered to children contains the following substances per 100 ml, with the energy level being 330 kJ. Protein 2.0 g, consisting of 1.9 g of milk proteins and 1.0 g of preparation according to Example 1.

Carbohydrates 7.8 g, consisting of 1.6 g of maltodextrins and 6.0 g of lactose, the remainder amounting to 0.2 g.

Lipids 4.4 g, consisting of 1.8 g of saturated fatty acids, 0.6 g of milk fat, 1.6 g of mono-unsaturated fatty acids, 0.65 g of polyon-saturated fatty acids and 0.35 g of glycerol as triglyceride.

Minerals, spore elements and vitamins were present in amounts which are normally recommended for a composition for children of up to 6 months. The composition according to Example 4 can be used for reducing the significant occurrence of diarrhoea (possible reduction of the dry solid matter content) and significantly reducing the concentration of pathogens in faeces.

Example 5

For treatment of the skin, a soft cream consisting of the following constituents was composed:

4 wt.% of lecithin, 2 wt.% of glycerol, 0,5 wt.% of protein according to Example 1, 2 wt.% of emul gator mush and furthermore 2 wt.% of cetylalcohol, 4 wt.% of isopropyl myristate, 1 wt.% of dimethiocon, 1 wt.% of squalane, 0.7 wt.% of preservative, 0,2 wt.% of perfume complemented with water to 100 wt.%.

Claims

1. A pharmaceutical product or food supplement which contains a preparation from blood plasma as the active component, characterized in that the preparation contains a larger amount of complement system (CS) and/or antibodies (immunoglobulins) than is normally present.

2. A pharmaceutical product or food supplement according to claim 1, characterized in that the preparation contains a larger amount of complement system (CS) than is normally present.

3. A pharmaceutical product or food supplement according to claim 1, characterized in that the preparation has been prepared from hyperimmune animal plasma.

4. A pharmaceutical product or food supplement according to claim 3, characterized in that said animal plasma is porcine plasma.

5. A pharmaceutical product or food supplement according to claims 3 - 4, characterized in that said plasma has been dried by spray- drying.

6. A pharmaceutical product or food supplement according to claim 1, characterized in that said preparation contains one or more growth factors (GF)

7. A pharmaceutical product or food supplement according to claim 6, characterized in that said growth factor is insulin-like growth factor (IGF).

8. A pharmaceutical product or food supplement according to claim 1, characterized in that immunoglobulin is present in an amount of

15 - 25 wt.% of the amount of solid matter of the preparation.

9. A pharmaceutical product or food supplement according to claim 1, characterized in that protein is present in an amount of more than 60 wt.% of the amount of solid matter.

10. A pharmaceutical product or food supplement according to claims 1 - 9, characterized in that 0.001 - 0.02 g plasma-protein per kilogram of body weight can be ingested per day.

11. A pharmaceutical product in a form which is suitable for medical application, characterized in that said product contains an active component according to claims 1 - 10.

12. A food supplement in solid or liquid form, characterized in that said food supplement contains an active component according to claims 1 - 13.

13. A product for the preparation of a pharmaceutical product or food supplement from blood plasma, characterized in that the amount of complement system (CS) is higher than 2.6 g per 100 g of product.

14. A product according to claim 13, characterized in that the amount of CS is higher than 3.25 g per 100 g of product.

15. A product according to claims 13 - 14, characterized in that the amount of CS is higher than 4 g per 100 g of product.

16. A pharmaceutical product or food supplement, characterized in that said pharmaceutical product or food supplement has been prepared on the basis of a product defined in claims 13 - 15.