WO1997000671A2 - Composition for preparing finely dispersed systems and process for preparing the same - Google Patents

Abstract

A composition contains at least one phospholipid, at least one bile acid and/or at least one bile acid derivative, ethanol and water. This composition is useful for preparing finely dispersed systems that remain stable even after a long storage time. The finely dispersed systems are particularly useful for preparing pharmaceutical or cosmetic compositions and have biological activity although they do not contain a usual pharmaceutical active substance.

Description

Composition for the production of finely dispersed systems and process for their production

The present invention relates to a new composition which is particularly suitable for the production of finely dispersed systems, and to a process for their production and their use.

The prior art contains numerous publications which prove that the effectiveness of, in particular, amphiphilic and lipophilic active pharmaceutical substances can be increased (ie better bioavailability is achieved, a lower dosage is required as a result and a lower toxicity is the result) is) when such active substances are incorporated into finely dispersed vesicular or particulate systems. Nevertheless, only a few remedies based on finely dispersed galenics are on the market to this day. The reason for this may on the one hand be seen in the fact that the development of such a product, among other things due to official requirements, takes many years, on the other hand the previously known preparations are not stable over the long term, which means that they are not suitable for marketing Question come.

Self-functionality, i.e. The biological effect of the finely dispersed systems, without containing an active ingredient, has so far been given no importance and has therefore not been investigated.

No compositions are known from the prior art which enable the production of finely dispersed, liquid systems with a long-term stability required for marketing. Liquid systems are understood to mean those with a viscosity of 0.9 to 100 mPa 's.

The present invention is therefore based on the technical problem of specifying a composition which the manufacturers The provision of long-term stable finely dispersed systems is made possible, the composition at the same time having its own functionality, that is to say having cosmetic or therapeutic properties, without containing other conventional pharmaceutical or cosmetic active ingredients.

This problem was solved according to the invention by a composition comprising the following components:

a) at least one phospholipid, b) at least one bile acid and / or at least one of its derivatives, c) ethanol and d) water.

Nanodisperse lipid / water formulations, preferably nanoemulsions, nano-colloids and liposomes, which are stable over a long period of time and have their own functional effectiveness in vivo and in vitro, can be prepared in a simple manner from commercially available devices from the composition mentioned. in particular in the cosmetic and pharmaceutical sector. The composition can be easily loaded with various active ingredients, the composition giving the active ingredients increased stability and effectiveness.

The phospholipid of the composition is preferably a phosphatidylcholine of pharmaceutical quality, the aliphatic residues of the phospholipid preferably comprising 12-22 carbon atoms with up to 4 cis double bonds, unbranched aliphatic residues being particularly preferred. A phospholipid in which the aliphatic radicals comprise 16-20 C atoms and a maximum of 3 cis double bonds is particularly preferred, as in natural vegetable lecithins, for example soybean lecithin, the aliphatic radicals R 1 and R _{2 being the} same or different Number of C- Can have atoms. The phospholipid can also be of natural, semi-synthetic or fully synthetic origin.

The bile acid and / or its derivatives can also be of natural, semi-synthetic or fully synthetic origin. Preferred derivatives are salts of bile acid, for example sodium salts.

Cholic acid, deoxycholic acid, glycolocholic acid, taurocholic acid, taurodeoxycholic acid, ursocholic acid, chenoxycholic acid are particularly preferred as bile acid. Preferred derivatives are sodium cholate, sodium deoxycholate, sodium glycocholate, sodium taurocholate, sodium taurodeoxycholate, sodium urocholate and sodium chenoxycholate.

96% ethanol according to DAB 10 is preferably used as ethanol. Ethanol appears to be essential for achieving long-term stability of the finely dispersed systems over a period of more than 18 months.

In comparative experiments, ethanol-free preparations which otherwise correspond to the composition according to the invention have proven to be physically unstable. Freshly prepared, ethanol-free preparations were also ineffective in vivo.

The composition according to the invention is preferably provided as a concentrate in which the phospholipid is contained in a concentration of 5 to 250 mg / ml, preferably 40 to 200 mg / ml and the bile acid and / or its derivative (s) is (are) contained in a molar ratio of 2 to 10 to 1 (lipid to cholate). Ethanol is preferably contained in a ratio of between 1: 2 and 1: 0.5 to the phospholipid (V / G), particularly preferably in a ratio of 1: 1.

The ethanol content in the stated ratio to the phospholipid used (volume ethanol / weight phosphorus) pholipid) gives the composition a particular stability and also facilitates the production process of the finely dispersed systems, and furthermore this ethanol content is of particular advantage for the intended effectiveness.

The concentrate according to the invention is diluted for use in a ratio of 1: 1 to 1: 200, preferably 1: 1 to 1:40 and particularly preferably 1: 5 to 1:20. Water or salt solutions, preferably those with 0.1 to 0.9% by weight of sodium chloride, serve as the diluent.

Particularly stable systems are achieved with a composition which contains 100 g of phospholipid, 0.2 g of tocopherol, 100 ml of 96% ethanol and 13.4 g of sodium cholate per liter of aqueous solution.

The composition is preferably used as a finely dispersed system based on lipid mixtures, micelles, mixed micelles, reverse micelles, uni-, oligo- or multilamellar liposomes, nanoemulsions, nanoparticles or nanocolloids. “Finely disperse systems” are understood here to mean systems with particles whose diameter is approximately 4 to 500 nm. The diameter of the particles is preferably less than 250 nm, very particularly preferably less than 100 nm. The vesicular systems have been best studied.

The composition preferably contains a pharmaceutical and / or cosmetic auxiliary, but long-term stability of the finely dispersed systems over several years can also be achieved without these auxiliary agents. In order to further expand the stability of the finely dispersed systems, antioxidants, for example, can be added to the composition if an unsaturated phosphatidylcholine is used as the phospholipid. Tocopherol is usually added as an antioxidant. From a medical point of view, such as, for example, achieving isotonia and from For reasons of osmolarity, salts, in particular table salt or buffer substances, can also be added, as described, for example, in EP-A-0056781, DE-A-4026833, DE-A-4026834, EP-A-0475160, EP-A-0613685 and WO94 / 01089. In addition to antioxidants, salts and buffer substances, cosurfactants such as, for example, polysorbate or free fatty acids, thickening agents and also preservatives are preferred as auxiliaries, as are customarily used in pharmaceuticals and / or cosmetics.

The composition is particularly suitable for the production of a pharmaceutical preparation. As stated above, the composition as such has biological activity which makes it appear particularly useful for use in the therapeutic field.

The inherent properties of the active ingredient-free composition make it particularly suitable for the treatment of bacterial and viral infections (as well as AIDS), for the treatment of cancer, Alzheimer's disease, Crohn's disease and ulcerative colitis, asthma, bronchitis, high blood pressure and also hypoxia. Furthermore, the pharmaceutical preparation according to the invention has been used effectively for the treatment of skin diseases and the treatment of the dry eye and the dry mouth. When applied topically to the skin and to the eyelids, it was found that non-oily formulations for the therapy of skin allergies and for the therapy of the dry eye were significantly effective. The optional addition of vitamin C also showed an antiviral and anticancerogenic activity.

One of the effects of the finely dispersed system can probably be attributed to the fact that in vivo application (orally or systemically, for example in IV or IV), the noxious substances are "thinned out": by supplying membranes which, although artificially produced, are made from natural phospholi - piden exist and therefore non-toxic and completely meta- Can be polarized, in the form of finely dispersed systems, the total membrane surface is increased in body fluids. The ability of these membranes to bind toxins, histamine, metabolite, pollutants, even oxygen in vivo and to remove them rapidly renally or bilaterally from the body relieves the blood cells and leads to a passive, purely physicochemical detoxification and thus a reduction of the noxious substances.

The intrinsic functionality of the systems can be strengthened in a synergistic manner by adding and / or storing active substances. The increase in activity through the combination of the finely dispersed system and the active ingredient is presumably due to the fact that the finely dispersed system disperses the active ingredient particularly finely in vivo in cell membranes and thus increases its effectiveness.

In a preferred embodiment, the pharmaceutical preparation according to the invention contains vitamin C or one of its derivatives. Vitamin C is usually used in a concentration of 0.0002 to 120 mg / ml.

This concentration range is so large because completely different application routes are possible, for example topically on the skin (occlusive, but also only sprayed on), orally, IV, IV, intralymphatic and inhalation, and as a test kit in vitro. Based on vesicular systems with 10% phospholipid, the concentration of vitamin C is preferably between 1 and 100 mg / ml.

In a further embodiment of a pharmaceutical preparation according to the invention, the composition is incorporated into gelatin, for example hard or soft gelatin capsules.

It has surprisingly been found that the finely dispersed system, in particular in the form of liposomes, the ATP level in several experimental cell culture systems. Furthermore, it has surprisingly been found that the composition can lower the cAMP level of isolated rat cortex. These surprising findings enable the composition according to the invention to be used for the prophylaxis and / or therapy of diseases in which it is desirable to lower the ATP and / or cAMP level. These diseases include, for example, cancer and Alzheimer's disease. In connection with cancer, it has been found that cancer cells have an elevated cAMP level and that lowering the cAMP level can eliminate the malignant phenotype. Furthermore, it was found in Alzheimer's disease that lowering the ATP level leads to a relief of the symptoms of the disease.

Furthermore, the preparation according to the invention, as a finely disperse system, in particular in the form of liposomes, shows an increased oxygen consumption and a significant increase in temperature in a lymphocyte culture. Furthermore, it was found that in vitro oxygen radical formation (oxonium ion formation) occurs, the release of such radicals being further increased by the addition of vitamin C.

By adding or storing known active ingredients (e.g. against pain or high blood pressure), their effect can be enhanced in a synergistic manner, and anti-inflammatory properties also occur.

This property of the preparation according to the invention also allows it to be used as an immunostimulating agent.

In addition, the preparation according to the invention shows a significant dose-dependent toxicity to leukemia cells (HUT78, L578Y), which could be increased synergistically by vitamin C. When testing with peripheral lymphocytes, the Toxicity of the composition decreased significantly. This opens the way for the composition according to the invention for the prophylaxis and / or therapy of cancer diseases (see also Example 6) and for the prophylaxis and / or therapy of monoclonal, polyclonal or malignant gammopathies, primary, secondary or combined immunodeficiency diseases, autoim - to use munopolyendocrinopathies and autoimmune diseases. The autoimmune diseases can be organ-specific, for example pernicious anemia, chronic gastritis, juvenile diabetes mellitus, and non-organ-specific diseases, such as diseases of the rheumatic type (eg rheumatoid arthritis, scleroderma) or mixed - Or transitional forms such as autoimmune haematolytic anemia, primary biliary cirrhosis, chronic aggressive hepatitis, ulcerative colitis, Sjogren's syndrome and multiple sclerosis.

In addition to this suitability for the production of pharmaceutical preparations, the composition according to the invention is also suitable for the production of cosmetic preparations, such preparations being particularly suitable for "simple" skin protection and normal skin care. Cosmetic preparations are also suitable for incorporation into gelatin.

Further substances, such as dyes, can also be added to the composition according to the invention. In the case of these compositions with dyes attached to or embedded in the particles of the finely dispersed system, in particular textiles, leather or paper can be dyed or conditioned.

In a further preferred embodiment, the composition according to the invention is added to the dialysis liquid usually used in kidney dialyzers. The effectiveness of blood purification and detoxification has been observed to increase significantly. The reason the rapid and effective adsorption of amphiphilic metabolites onto the particles of the finely dispersed system, in particular the liposomes, should be found for this.

Finely dispersed systems can be produced from the composition according to the invention by adding mechanical energy, such as, for example, by optimized mixing in static mixers, pressure filtration, stirring, homogenization or filtration. Commercially available devices can be used for this, such as static mixing systems or Gaulin homogenizers or a microfluidizer. If more than 100 mg / ml of oil, based on the composition, is added in these processes, the process leads to nanoemulsions or nanocolloids, and without addition of oils or oil concentrations of <30 mg / ml, based on the composition the procedure to liposomes.

All such "pharmaceutical" oils can be used as such oils, e.g. Miglyol, jojoba oil, borage oil, castor oil, safflower oil, paraffin oils etc., and also essential oils such as 01-bas, tea tree oil, peppermint oil etc.

example 1

10 g soy lecithin (phosphatidylcholine content 97%) are dissolved in 10 ml 96% ethanol. 650 mg table salt and 1.3 g sodium cholate are injected into 180 ml water. solved. Both solutions are combined and sterile filtered at 3.5 bar. The resulting finely dispersed system is brought to the desired application concentration by dilution with isotonic saline or even with water.

Example 2

10 g of soy lecithin as in Example 1 are 96% in 10 ml Ethanol dissolved. 650 mg of common salt and 1.3 g of sodium cholate are dissolved in 80 ml of water. Both solutions are combined and homogenized in a household mixer, eg SG stick. The resulting finely dispersed concentrate is brought to the desired application concentration by dilution with isotonic saline solution or even only with water.

Example 3

500 g soy lecithin as in Example 1 are 96% in 500 ml

Ethanol dissolved. 32.5 g of sodium chloride and 65 g of sodium cholate are dissolved in 4 l of water. Both solutions are combined and in a standard agitator, e.g. Stephan UMC 12, homogenized. The resulting finely dispersed system is diluted to the desired application concentration as in Example 1.

The resulting particles have an average diameter of less than 200 nm (measured with a light scattering device Coulter N4SD). Their diameter no longer changes due to the thinning steps. Dilutions 1: 1, 1: 5, 1:10, 1:20, 1:50, 1: 100, 1: 200, 1: 400 were checked.

Example 4

If an active ingredient is to be stored in the finely dispersed system, there are in principle two options which depend solely on the solubility properties of the active ingredient in question:

Water-soluble active ingredients are dissolved together with the sodium cholate in the aqueous phase before it is combined with the ethanolic lipid solution. Lipophilic active ingredients are dissolved in the ethanol together with the lipid. Become oils dissolved in the ethanolic lipid solution before combining with the aqueous phase or added to the ready system and stirred with the system a second time.

Of course, several different active ingredients can be incorporated into such a system at the same time.

If the solubility properties of active ingredients to be stored in the finely dispersed system are poor, the solubility can be improved by adding co-surfactants, e.g. Polysorbate 80, Arlasolve, free fatty acid such as oleic acid etc., and / or by small additions of other alcohols, e.g. Isopropanol, glycol or propylene glycol.

If an active ingredient is to be added to the finely dispersed system, its solubility properties are predissolved in additional ethanol or aqueous phase, added to the finished finely dispersed system and incorporated by stirring.

Example 5

Do active substances impair the finely dispersed system in the basic formulation, e.g. Oxidizing agent, the desired storage can take place in a two-chamber system from which the active substance is mixed with the finely dispersed system only shortly before use.

Example 6

The patient JLSM, 47 years old, was diagnosed with duodenal ulcer at the age of 17 years. During a routine check in October 1990, a prepoly ulcer was also found. Both were more or less controlled. Routine control in November 1994 showed the endoscopic biopsy is transformed into a little differentiated carcinoma. A surgical treatment was then recommended and carried out. 15 days before the operation, the patient received 30 ml of liposomes containing ascorbate daily. The tissue removed during the operation was examined histologically. Diagnosis: Ulcer pepticum with fibrinoid necrosis at the base and regenerative changes on the edge. No evidence of atypia, no deterioration.

Claims

claims 1. Composition comprising the following components: (a) at least one phospholipid, (b) at least one bile acid and / or at least one derivative thereof, (c) ethanol and (d) water. 2. Composition according to claim 1, characterized in that (a) the phospholipid has the following general formula: R _! -C-0-CH ₂ R ₂ -C-0-CH O OH ₂ C-0-P-0-CH ₂ -CH ₂ -N (CH ₃ ) ₃ O wherein R 1 and R ₂ each represent an aliphatic radical having 12 to 22 carbon atoms and having up to 4 cis double bonds, the aliphatic radicals preferably being unbranched. 3. Composition according to claim 1 or 2, characterized gekenn¬ characterized in that the bile acid and / or its derivative is selected from cholic acid, deoxycholic acid, glycocholic acid, taurocholic acid, taurodeoxycholic acid, ursocholic acid and chenoxycholic acid or their salts, preferably their group comprising sodium salts ( are) . 4. Composition according to one of claims 1 to 3, characterized in that it contains: (a) the phospholipid in a concentration of 5 to 250 mg / ml, preferably 40-200 mg / ml, (b) the bile acid and / or its derivative (s) in a molar ratio of 2 to 10 to 1 (lipid to cholate), (c) Ethanol in a ratio of preferably 1: 1 (v / w) to the phospholipid. 5. Composition according to one of claims 1 to 3, characterized in that by diluting the composition according to claim 4 in a ratio of 1: 1 to 1: 200, preferably 1: 1 to 1:40 and particularly preferably 1: 5 to 1: 20 is available. 6. Composition according to claim 5, characterized in that the diluent is water or a salt solution, preferably 0.1 to 0.9% saline. 7. Composition according to at least one of claims 1 to 6, characterized in that the composition as a finely disperse system based on lipid mixtures, micelles, mixed micelles, reverse micelles, uni-, oligo- or multilamellar liposomes, nanoemulsions, nanoparticles or nanocolloids with a diameter of preferably less than 250 nm, particularly preferably less than 100 nm, is present. 8. The composition according to at least one of claims 1 to 7, characterized in that it further contains at least one pharmaceutical and / or cosmetic auxiliary. 9. Pharmaceutical preparation containing the composition according to one of claims 1 to 8. 10. Pharmaceutical preparation according to claim 9, characterized in that it further contains at least one pharmaceutically active ingredient. 11. Pharmaceutical preparation according to claim 9, characterized in that it contains vitamin C and / or one of its derivatives. 12. Pharmaceutical preparation according to one of claims 9 to 11, characterized in that it contains the composition incorporated in gelatin, such as hard or soft gelatine capsules. 13. Use of the pharmaceutical preparation according to one of claims 9 to 12 for lowering the ATP and / or cAMP level. 14. Use of the pharmaceutical preparation according to one of claims 9 to 12 for the prophylaxis and / or therapy of cancer. 15. Use of the pharmaceutical preparation according to one of claims 9 to 12 for immunostimulation. 16. Use of the pharmaceutical preparation according to one of claims 9 to 12 for the prophylaxis and / or therapy of gammopathies, immune deficiency diseases and autoimmune diseases. 17. Cosmetic preparation containing the composition according to any one of claims 1 to 8. 18. Cosmetic preparation according to claim 17, characterized in that it incorporates the composition in gelatin, e.g. Contains hard or soft gelatin capsules. 19. The composition according to any one of claims 1 to 8, characterized in that it contains other substances, such as dyes, attached or incorporated. 20. Use of the composition according to claim 19 for dyeing and / or conditioning textiles, leather or paper. 21. Use of the composition according to one of claims 1 to 8 for the production of a finely dispersed system. 22. Use of the composition according to one of claims 1 to 8 as an additive to the dialysis fluid in hemodialysis. 23. A method for producing a finely dispersed system comprising the composition according to at least one of claims 1 to 8, comprising the steps (a) pressure filtration, preferably sterile filtration, the composition and / or (b) Mixing or stirring or homogenizing the composition with commercially available devices.