MXPA00000545A - Lipid-based immune modulator composition - Google Patents

Abstract

A pharmaceutical composition for activating the immune system comprises a lipid preparation derived from a natural source enriched to comprise at least about 10%phosphatidic acid.

Description

Nomodulatory inm composition based on lipid FIELD AND BACKGROUND OF THE INVENTION The present invention concerns a composition and method for the activation of the immune system. The composition of the present invention is particularly useful for the activation of the immune system in a stress situation where the activity of the immune system decreases. A particular example of an induced stress that decreases in the activity of the immune system is in the case of smoking cessation. There are many clinical conditions where the activity of the immune system decreases, therefore making the individual more susceptible to opportunistic infections. It is also known that stress, such as that resulting from injuries, rehabilitation, addiction to drugs, alcohol or smoking, can result in a decrease in the activity of the immune system, in a matter of minutes. For example, strong infections that occur as a result of severe injury have been documented. The aim of the present invention is to provide a composition for improving the activity of the immune system REF .: 32554 to make it more active, particularly useful in stressed individuals.

SUMMARY OF THE INVENTION In accordance with the invention, it was surprisingly found that the preparation of a natural lipid enriched with phosphatidic acid (PA) is capable of activating the immune system. This activity of the PA-enriched lipid preparation, which was found to be associated with decreased activity of the immune system, was tested in individuals undergoing cessation smoking treatment. In the following, the use will be made with various terms, these terms and their meanings in the context of the invention, are as follows: Preparation of PA-enriched lxpide (PA-E-LP) - a lipid preparation comprising at least 10% (w / w) of PA, preferably within the range of about 20% -75% of PA, outside the total lipid content of the composition. (The indication of "%" concentration given above and below denotes the number of units of weight of an ingredient per 100 units of weight of the entire composition (w / w)).

PA-E-LP Natural - a PA-E-EP derived from a preparation of the natural lipid, for example, a preparation of the phospholipid derived from plants, from animal tissue, or any combination thereof. Thus, a natural phospholipid preparation can typically be derived from soybeans, egg yolks or animal sera. The PA-E-EP is prepared from a preparation of the natural phospholipid, typically by an enzymatic process. In a natural PA-E-EP, the rest of the lipids consists primarily of phospholipids although in small amounts, for example 0.1-10%, of other lipophilic substances, such as cholesterol, fatty acids, etc., may also be included. In the preparation. Stressed individuals - individuals exposed to a situation or condition such as anxiety, physical injury, cessation of drug use, use of alcohol, smoking, etc. that increase stress. The present invention provides, by one of its aspects, a pharmaceutical composition for the activation of the immune system, comprising a preparation of the lipid derived from a natural source enriched to comprise at least about 10% of PA, preferably at least of almost 20% of PA and more desirably above 50% PA. Typically the concentration of PA, outside the total lipid ingredient, would not exceed 75%. The composition of the invention is particularly useful in the activation of the immune system under conditions of related stress that involve the reduction of the activity of the immune system. Examples of such stress conditions are a variety of injuries, cessation of drug use, alcohol, smoking, etc. The present invention further provides a method for activating the immune system in an individual in need, comprising administering an effective amount of PA-E-LP to the individual, particularly natural PA-E-LP. The invention still further provides the use of the aforesaid PA-E-LP and particularly of natural PA-E-LP, for the preparation of a pharmaceutical composition for the activation of the immune system. The term "effective amount" should be understood as an amount of an ingredient sufficient to produce a desired therapeutic effect. For example, administration of an effective amount of the pharmaceutical composition of the invention to an individual results in an increase in the level of activation of the immune system. The level of activation can be measured, for example, by the blood plasma level of gaiti a-interferon (? -IFN), which can in this case, serve as a measure to determine the effective amount. The term "activation of the immune system" or similar term should be understood as a reference for increasing the level of activity of immune cells in combatted pathogens or pathological processes, increasing the level of secretion or production of several cytokines thereby assisting such activities of immune cells, etc. The pharmaceutical composition of the invention can typically be administered orally although this can also be formulated by local or parenteral administration. For oral administration the composition may comprise various flavoring agents, edible colors, etc. In addition, the composition for oral use can also be encapsulated, for example, in a gelatin-filled capsule. The composition may comprise a variety of pharmaceutically acceptable carriers, diluents or excipients, which may be chosen based on the desired mode of administration of the composition. For parenteral administration the composition will typically be injected intravenously (I.V.). Such a parenteral composition may comprise for example soy bean triglycerides, egg phospholipids, PA prepared according to the invention, glycerol and distilled water. A local composition may be in the form of a gel or an ointment and may therefore comprise various additives known per se to allow the compositions to obtain such a physical form (eg, a gelatinous agent). Natural PA-E-LP is preferably obtained from a preparation of the natural phospholipid by enzymatic treatment using a synthetic or natural source comprising the enzyme phospholipase-D. The preparation of the natural phospholipid can be of vegetable origin, it can be of animal origin, or a combination thereof. Typical examples of the natural phospholipid preparations useful for the preparation of natural PA-E-LP of the invention are soy lecithin, egg yolk and animal serum phospholipids. Examples of phospholipase-D sources are peanuts, typically ground peanuts or a fraction of phospholipase-D derived from that. Phospholipase-D or a source of phospholipase-D is added in an amount and for a time sufficient to hydrolyze at least about 10% or preferably about 25% and more preferably about 50% of the phospholipids to produce PA-E- LP. The present invention will now be illustrated in the following non-limiting examples with occasional reference to the appended drawings.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic representation describing the preparation of the PA-enriched mixture of the invention. As you can see in the figure, the ratio of the initial materials (peanuts, soy lecithin and water) is 1: 1: 7.5, respectively. Fig- 2 is a schematic representation describing the separation of the lipid phase from the PA mixture prepared by the reaction shown in Fig. 1. Fig. 3 is a schematic representation describing the preparation of the PA mixture of the invention. prepared as described in Figs. 1 and 2 in a suitable way to fill the containers. Fig. 4 is a schematic representation describing the preparation of tablets of the PA mixture prepared as described in Figs. 1 and 2.

Fig. 5 shows the profile of formation of the PA during an enzymatic hydrolysis of the phospholipids of the soybeans. (The results are present as% of the entire composition).

Example 1: Preparation of the PA mixture (a) Preparation of a reaction product Fig. 1 schematically describes the method of preparing a PA mixture according to the invention. As you can see, the initial materials are as follows: 1. 150 grams of fresh peanut kernels as an origin of Phospholipase D. 2. 150 grams of granulated soy lecithin as an origin of soybean foepholipids. 3. 1000 mL to be added to H20. The above-mentioned starting materials were mixed in a meat mixer, homogenized and then the pH of the mixture was adjusted to a pH of 5.3-5.4 by the addition of calcium lactate, sorbate, benzoate, ascorbic acid, citric acid and an anti-oxidant (each in the amount described in the figure). The mixture was then reacted under continuous mixing for four hours at 36-38 ° C after which the pH was again adjusted to a pH of 3.9 by the addition of citric acid and sugar in amounts described in the figure. The mixture was then stored at 4 ° C overnight. (b) Separation of the lipid fraction The reaction mixture obtained by the above procedure is then further separated by a procedure which is schematically described in Fig. 2. As seen in the figure, the reaction product is subject to three main separations (indicated as separations I, II and III in the figure) where, generally, in each separation the aqueous phase of the mixture is removed from the bottom until finally, the phase of the separated lipid is collected and stored in a freezer ( -18 ° C). (c) Preparation of the lipid composition for filling the containers The mixture of the lipid obtained by the above separation was subjected to several additional procedures by which it was prepared for the filling of the containers. As seen in Fig. 3, the mixture of the lipid obtained by the above separation process was first melted and then treated by heating, cooling, homogenization and mixing as described in Fig. 3 to form a composition suitable for filling of the containers. Typically the size of the containers are half a liter and the containers are then stored at 4 ° C. (d) Preparation of PA containing tablets The PA mixture obtained by the method described in Examples 2 (a) -2 (c) above and shown in Figs. 1-3 can also be processed into tablets. The method for the preparation of the tablets comprising the PA mixture is shown in Fig. 4. As seen in the figure, generally the phospholipid paste obtained by the above-mentioned process is first thawed and then heated and the pH of the mixture is adjusted to a high pH of almost 9. Following the addition of zinc acetate to the mixture, a large sediment appears which, after washing and drying, is formulated into tablets. (e) PA formation profile The PA formation profile is described in Fig. 5. As can be seen, after almost 24 hours, more than 80% of the soybean phospholipid was hydrolysed producing a preparation of PA enriched.

Example 2: Effect of a PA-E-LP composition on the increased levels of gamma-interferon in individuals undergoing smoking cessation treatment (a) Study design 35 cigarette smokers of which from 20 to 40 cigarettes at day, and those who voluntarily enrolled in the study, were randomly assigned into two groups: a group, consisting of 16 individuals, received a daily dose of 12 grams (taken orally) of the composition of PA-E-LP prepared from according to Example 1, which contains 50% PA; and the second group, which serves as a control, received a placebo treatment. Individuals in both groups stopped smoking during the study. Concentrations of? -IFN in plasma were measured in individuals of both groups, before and after cessation of smoking. The level of? -IFN was measured by the use of the QUANPIKINE ™ Package (R &D Systems Inc., USA), (b) Results The concentrations of? -IFN in the two groups, before and after the treatment are shown in the next Table 1.

Table 1 Concentrations of? -IFN from Plasma H It can be seen from the above results, the pink concentration of? -IFN of the blood plasma in the group receiving the PA-E-LP, while the? -IFN concentration of the plasma in the placebo group was lower than its concentration before treatment. It should be noted, that the? -IFN levels of the blood plasma are an indication of the activation state of the immune system.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (5)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. A pharmaceutical composition for the activation of the immune system, characterized in that it comprises a preparation of lipid derived from a natural source enriched to comprise at least about 10% of PA.

2. A pharmaceutical composition according to claim 1, characterized in that it comprises at least about 20% of PA.

3. A pharmaceutical composition according to claim 1, characterized in that it comprises at least about 50% of PA.

4. A pharmaceutical composition according to any of claims 1-3, characterized in that the preparation of the lipid is derived from a source of natural lipid.

5. A pharmaceutical composition according to any of claims 1-4, characterized in that it activates the immune system in stressed individuals. A pharmaceutical composition according to claim 5, characterized in that it activates the immune system in stressed individuals as a result of the cessation of the use of drugs, alcohol or smoking. A pharmaceutical composition according to any of claims 1-6, characterized in that it activates the immune system in a stressed individual, wherein said activation results in an increase in the levels of? -IFN in the plasma of the treated individual. A pharmaceutical composition according to any of claims 1-7, characterized in that it is formulated for oral administration. A method for activating the immune system in an individual, characterized in that it comprises administering to the individual an effective amount of a PA-enriched lipid preparation. A method according to claim 9, characterized in that the preparation of the lipid is derived from a natural lipid preparation. A method according to claims 8 and 9, characterized in that said preparation is administered orally. The use of a lipid preparation derived from a natural source enriched to comprise at least about 10% PA. The use according to claim 12, characterized in that the lipid preparation comprises at least about 20% of PA. The use according to claim 13, characterized in that the lipid preparation comprises at least about 50% of PA. The use according to any of claims 12-14, characterized in that the lipid preparation is derived from a source of natural lipid. The use according to any of claims 12-15, for the preparation of a pharmaceutical composition for the activation of the immune system in stressed individuals. The use according to claim 16, characterized in that the individuals are stressed as a result of a smoking cessation treatment.