SE517678C2 - Lipophilic carrier preparations with a continuous lipid phase - Google Patents

Abstract

FIELD: pharmaceutics. SUBSTANCE: in a lipophilic carrier containing polar lipid material and non-polar lipid, the former is galactolipid containing at least 50% (advantageously 100%) of digalcatosyl(diacyl)glycerols along with other polar lipids. Carrier may contain polar solvent, for instance, an aqueous solution. It may as well be in the form of reverse vesicles or reverse micelles, or water-in-oil emulsion. Carrier can be used as auxiliary component in pharmaceutical, cosmetic, or food products. Pharmaceutical compositions contain such carriers in combination with bioactive compounds. EFFECT: improved biological compatibility of carrier-containing products. 12 cl, 3 tbl, 8 ex

Description

517 678 2 lipid such as soybean phospholipids (soy lecithin). These emulsifiers consist of mixtures of phospholipid classes, such as phosphatidylcholine and phosphatidylethanolamine, which are zwitterionic, and phosphatidylinositol, which is ionic. It is well known that these lecithin emulsifiers are the most widely used natural lipids in the production of stable food emulsions on an industrial scale. It is also well known that such emulsions have disadvantages and problems associated with the emulsifier being lecithins.

Prior Art There are only a few examples reported of lipophilic preparations having a continuous lipid phase.

WO 92/05771 discloses a lipid particle forming matrix of at least two lipid components; one is non-polar and another amphipathic and polar. This particle-forming matrix, which may contain bioactive materials, spontaneously forms discrete lipid particles when interacting with aqueous systems. The amphipathic and polar lipid components are said to be bilayer-forming and are selected from phospholipids such as phosphatidylcholine; the non-polar lipids are mono-, di- or triglycerides.

Microemulsion gels containing lecithin, i.e. phosphatidylcholine, have been described and characterized by P. L. Luisi, see for example D. Capitani et al, Langmuir, 1993, vol 9, pages 685-689. In addition to the phospholipids, these gels are composed of a small amount of water and an organic solvent, such as alkanes, fatty acid esters and amines. They are also called organogels. The gels can be used as a matrix for transdermal transport of drugs.

The presence of inverted vesicles, counter structures to normal vesicles, in an oil was first reported by H. Kunieda, see for example H. Kunieda et al, Advanced Materials, 1992, vol 4, pages 291-293. Inverted vesicles constitute a dispersion of lamellar liquid crystal, which swells a considerable amount of oil, ie the vesicles consist of inverted bilayer structures. The inverted bilayers are normally composed of a mixture of hydrophilic and lipophilic amphiphiles.

Glycosylglycerides are a type of glycolipid that are well known constituents of plant cell membranes. Two types based on galactose are very common, monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), which make up up to 40% of the dry weight of the thylakoid membranes. The monoacyl compounds may also be present to some extent, ie monogalactosyl monoacylglycerol (MGMG) and digalactosyl monoacylglycerol (DGMG). To some extent, a sulfolipid, consisting of MGDG which contains -SCgH and is known as sulfokinovosyldiacylglycerol, may also be present.

SE 9400368-8 describes an industrially useful process for the preparation of glycosylglycerides from plants, preferably cereals, by means of extraction and chromatographic separations.

When examining the interaction between glycosylglycerides and non-polar lipids, such as mono-, di- and triglycerides, fatty alcohols and acids, sterols and sterol esters, possibly in combination with other polar lipids, such as phospholipids and sphingolipids, without water or with only small amounts of water , we have surprisingly found a behavior that makes such preparations suitable as lipophilic carriers for drugs, but also formulations for skin care, nutrition and food.

No pharmaceutical or other preparations or formulations based on glycosylglycerides have been previously known or realized in the art.

Description of the Invention The present invention provides lipid carriers based on glycosylglycerides, which constitute a nonionic lipid material, in combination with non-polar lipids. The polar main groups are carbohydrate moieties, which have completely different physicochemical properties compared to the phospholipids. The mechanism of incorporation is thus based on the interaction between the hydroxyl groups of the polar main group, i.e. the carbohydrate moiety, and the lipophilic chains of the glycosylglycerides and the non-polar component, and the compound to be incorporated.

The invention relates to a lipophilic carrier composition with a continuous lipid phase, which comprises a non-polar lipid in combination with a nonionic, polar lipid material, and optionally water, which is characterized in that the polar lipid material is a glycosyl glyceride of the general formula 517 678 4 R1 -o-cšn, Rz-o-CH Hzcl-o I (monosaccharide anzone) n-RB wherein R1 and RQ are independently saturated or unsaturated, linear or branched fatty acid residues having 0-24 carbon atoms and 0-6 double bonds; monosaccharide independently is a pentose or a hexose; n is 1-5; and R 3 is hydrogen or -SO 4; or a mixture thereof.

The glycosylglycerides are suitably a mixture of galactosylglycerides of the general formula R 1 -O-c 2 Z 2 R 2 -O-CH H 16-18, and 0-3 double bonds, and Ra is hydrogen or a galactose group.

Preferred examples of fatty acid residues R1 and RQ are naturally occurring fatty acyl groups, such as from the saturated acids palmitic acid (CuHuCO; 16: 0) and stearic acid (CUH§CO; 18: 0); from the monounsaturated acid oleic acid (CUH 2 CO; 18: 1); and from the polyunsaturated acids linoleic acid (CnHnCO; 18: 2); and linolenic acid (CUH 2 CO; 18: 3).

Preferred examples of monosaccharides are the pentoses ribose, arabinose, xylose and ribulose; the hexoses glucose, mannose, galactose, fructose and sorbos.

The specific amounts of the components of the glycosylglyceride material used in the examples below are not critical to the present invention. However, maximum benefits are achieved through a high level of digalactosyldiacylglycerols (DGDG), the major bilayer-forming components, in every conceivable application.

A preferred glycosylglyceride material therefore consists of 5G 67G mainly DGDG but also of monogalactosyldiacylglycerols (MGDG). Synthetic glycosylglycerides based on galactose or any other carbohydrate moiety, such as glucose, and natural glycosylglycerides, isolated from any source, based on carbohydrate moieties other than galactose, such as glucose, may be used in accordance with the invention.

The glycosylglyceride material can be extracted from almost any type of glycolipid-containing plant material. Preferred plant materials are seeds and grains from cereals and cereals, such as wheat, rye, oats, corn, rice, millet and sesame. Oatmeal as well as wheat gluten have a high lipid concentration and are therefore suitable for use in the production process.

There is no particular limitation on the non-polar lipid material. Vegetable oils, animal oils, synthetic oils, fatty acids, natural and synthetic glycerides, sterol esters, fatty alcohols, etc. can be mentioned as examples.

It was surprisingly found to be possible to obtain all types of lipophilic organized solutions by using glycosylglycerides. The incorporated active materials can form part of the organized solutions or be stabilized by them in the form of, for example, suspensions.

For example, a lipophilic carrier composition can be obtained by mixing the glycosylglycerides with a non-polar lipid, such as a triglyceride. The triglyceride can be selected from palm oil or natural oils with the same, relatively high content of solid fat or melting point range. Preferred and suitable triglycerides are palm oil fractions obtained by fractionating commercial palm oil in specific mixtures of triacylglycerols, based on the combination mainly palmitic, oleic and stearic acid esters of glycerol.

A lipophilic carrier preparation can also be obtained by mixing the glycosylglycerides with liquid triglyceride oils, such as medium chain triacylglycerol oil (MCT oil) and soybean oil.

Furthermore, this mixture of non-polar lipid-glycosylglyceride may contain increasing amounts of water or aqueous solution which may lead to the formation of inverted vesicles, inverted micelles and water-in-oil emulsions. Inverted vesicles are prepared by mixing a mixture of gly- o o -. ,. Cosylglycerides and a more polar amphiphile, such as lysophosphatidylcholine, in a ratio of 4: 1 by weight, are added to a triglyceride oil, preferably MCT oil. The total concentration of amphiphiles is less than 3% (w / w). A small amount of water or aqueous solution, less than 1% (w / w) of the total preparation, is then added. After ultrasonication, a fine dispersion of inverted vesicles is obtained.

At higher glycosylglyceride concentrations, but still low water content, 0.5-2% (w / w) of the total preparation, reverse micelles can be prepared. Inverted micelles, also called microemulsions, consist of water aggregates in an oil. Inverted micelles are thermodynamically stable. The shape and structure of the aggregates can lead to viscous systems, "microemulsion gels".

At still higher water content,> 5% (w / w) of the total preparation, water-in-oil emulsions are formed. These are two-phase systems, which consist of finely dispersed water droplets in an oil. Water-in-oil emulsions are thermodynamically unstable, but can be kinetically stable.

The aqueous solution may consist of pure water, buffer solution, saline, solutions of glucose, galactose, propylene glycol, polyethylene glycol, glycerol, etc. Furthermore, thickeners such as gelatin, agarose, carrageenan, methylcellulose and ethylhydroxyethylcellulose may be used.

The invention also relates to the use of a lipophilic carrier preparation as described, as a carrier for an active substance in a pharmaceutical or cosmetic product, food or agricultural product.

The invention also relates to a pharmaceutical composition comprising a lipophilic carrier composition as described, in combination with a bioactive substance.

The invention also relates to a pharmaceutical composition comprising a lipophilic carrier composition as described, wherein the non-polar lipid is an MCT oil, a palm oil fraction, or a bioactive substance.

The bioactive substance may be a lipophilic drug such as anticancer agents, antimicrobial and especially antifungal agents, immunosuppressive drugs such as cyclosporine, dermatologicals such as analgesics and antiphlogistic agents, antipruritic agents, disinfectants, astringents and adjuvants. hormones, psychotropic drugs, anesthetics and other drugs that are lipophilic and that can present formulation problems that can be solved through the use of glycosylglycerides. There are also many lipids such as fatty acids, mono-, di- and triacylglycerols, phospholipids, cholesterol esters and lipids of many other types which have a therapeutic effect in themselves and which can advantageously be formulated in the form of a lipophilic carrier, based on the glycosylglycerides. In this case, the bioactive substance is the non-polar lipid.

A pharmaceutical and cosmetic preparation can be prepared by melting a palm oil fraction in an open water bath at a temperature range of 40-70 ° C. The active ingredients and glycosylglycerides are weighed into a glass bottle. The molten palm oil fraction is transferred to the glass bottle and the mixture is dispersed with a mixer under high shear at about 1000 rpm and at a temperature range of 40-70 ° C for 2-4 minutes. The amount of water or aqueous solution required to form a topically applicable formulation is added and the formulation is thoroughly mixed with a rod.

The pharmaceutical composition may be formulated for oral, enteral, parenteral, rectal, vaginal, topical, ocular, nasal or aural administration to animals, especially mammals, including humans.

Topical skin care preparations can be roughly classified as medical topical skin care preparations and cosmetic preparations according to the way in which they are used.

Examples of medical topical skin care preparations are various ointments that contain one or more active ingredients. Ointments include both those containing an oil base and those containing an oil-in-water or water-in-oil base of the emulsion type.

When used as a cosmetic preparation, it is possible to mix, in addition to the essential ingredients, those routinely used as cosmetic ingredients such as oily substance, ultraviolet absorber, alcohol, chelating agent, pH regulator, antiseptic, thickener, pigment, perfume base and similar, if required in combination.

As cosmetics, one can formulate skin cosmetic preparations of various forms including, for example, emulsified cosmetics of the type water-17As7s 8 ten-in-oil or oil-in-water, creams, cosmetic emulsions, eau-de-toilette, oily cosmetics, lipsticks, foundation cream , skin cleansing preparations, hair lotions, hair-laying preparations, hair care preparations, hair growth stimulants, etc.

Water-in-oil emulsions, such as margarine and spreads, are prepared by conventional methods.

Brief Description of the Drawings Figure 1 shows a video-enhanced microscopic image of inverted vesicles of glycosylglycerides in MCT oil formed by ultrasonication according to Example 2, Composition A. The rod corresponds to 10 Fm.

Figure 2 shows a corresponding view of the inverted vesicles of composition B in Example 2.

Example A glycosylglyceride material, consisting mainly of dialactosyldiacylglycerol (DGDG) but also monogalactosyldiacylglycerol (MGDG), used in Examples 1-4 as a polar, nonionic lipid was prepared as follows: 200 kg of oatmeal (Kungsaldes, Sweden) and extracted with 1000 l of 95% ethanol at 70 ° C for 3 h in an extraction tank with stirring. The slurry was centrifuged while still hot and separated from solid particles. The liquid fraction was evaporated at a maximum of 60 ° C to give about 10 kg of extract in the form of a light brown oil.

The carbohydrates were removed as follows: 10 kg of the extract was thoroughly mixed with 10 l of hexane (polymerization grade hexane, distillation range 65-70 ° C, Shell) and 50 l of 70% aqueous ethanol. The mixture was separated into two phases overnight. The lower, more polar phase, which contained the carbohydrates, was discarded. The lipids are in the upper phase.

The upper phase, which amounted to about 20 l, was applied to a 30 cm x 20 cm stainless steel column (ID x'h) with a volume of about 14 l and containing 6.25 kg of silica gel (Matrex Silica Si, particle size). 20-45 mm, pore diameter 60 Å, from Amicon Corp., Danvers). The column was heated to 50 ° C and then washed with 30 L of a hexanesisopropanol mixture, 90:10, to remove any triglycerides. The glycosylglyceride material was then eluted from the column with 20 l of a hexanesisopropanol mixture, 60:40, to give a glycosylglyceride fraction. Evaporation of this fraction gave about 700 g of glycosylglycerides. The main lipid class was DGDG,> 70% (of the surface), according to analysis by high performance liquid chromatography.

The glycosylglycerides were then subjected to freeze-drying to give a free-flowing powder.

Example 1. Preparation of an anhydrous disulfiram formulation A parenteral formulation with disulfiram, an alcohol repellent, was prepared using the following ingredients: fl δ train weight 3 (w / w) Glycosylglycerides 20.0 Disulfiram (Sigma) 32.0 MCT oil (Karlshamns) 48.0 The ingredients were mixed using an Ultraturrax homogenizer at 2000 rpm for 15 minutes and at 3000 rpm for 5 minutes.

This gave a suspension containing a high amount of finely dispersed disulfiram particles, with a smooth and homogeneous consistency. The suspension showed excellent physical stability due to the presence of glycosylglycerides and no sedimentation could be observed when stored at room temperature. The viscosity of the suspension was relatively low and it was possible to apply the formulation through a syringe equipped with a thin (i.d. 1.0 mm) needle.

The formulation can be used for instillation into the duodenum in humans, where it can act as a depot for disulfiram which provides a prolonged alcohol deterrent effect.

Example 2. Preparation of inverted vesicles Inverted vesicles were prepared using the following ingredients: n a v u nu nu 517 678 :: = ... = - fš. Composition A Composition B Composition B 3 (w / w) 3 (w / w) Glycosylglycerides 1.65 1.56 Lysophosphatidylcholine 0.40 - from soybean (Karlshamns LipidTeknik) Phosphatidylinositol - 0.38 from soybean (Karlshamns LipidT) 0.56 0.64 MCT oil (Karlshamns) 97.39 97.42 After weighing the ingredients, the mixtures were sonicated in an ultrasonication bath for 1 hour at 30-40 ° C. The fine dispersions formed were stable for more than a week. The presence of large inverted vesicles was evaluated using a differential interference phase contrast microscope (Nikon X2F-NTF-21, Japan) with a video-amplified system (Argus 10, Hamamatsu Photonics Co., Japan), see Figures 1 and 2.

The inverse vesicles in this example are based on lipid ingredients suitable for use in pharmaceutical and cosmetic applications. Previously, reverse vesicles have been prepared by the use of phospholipids or synthetic surfactants in hydrocarbon oil, the latter constituents of which are normally too toxic for human use. Furthermore, the inverted vesicles of the present invention exhibit much better stability than previously reported for systems based on synthetic surfactants and hydrocarbon oil.

A reverse vesicle dispersion is an example of an organized solution in which bioactive materials, such as protein drugs such as interferons, and peptide hormones such as calcitonin or insulin, can be incorporated. Incorporation of a water-soluble protein drug or hormone into a triglyceride oil by reverse vesicles can facilitate the transport of the drug through lipophilic cell membranes. The drug molecules are located in the bilayers of the inverted vesicles which have a stabilizing effect on the drug. In particular, the drug can be protected from degradation in the intestine by oral administration. 517 ml Example 3. Preparation of reverse micelle; A highly viscous reverse micellar gel (organogel, microemulsion gel) was prepared as follows: Ingredient:% (w / w) Glycosylglycerides 17.9 MCT oil (Karlshamns) 81.3 Water 0.8 The glycosylglycerides and MCT oil were mixed together to a homogeneous dispersion. Water was added dropwise to the dispersion with continuous magnetic stirring. A clear gel formed almost immediately and maintained a high viscosity for over a month.

Example 4. Preparation of a water-in-oil emulsion A water-in-oil emulsion was prepared with the following ingredients: Component 3 (weight weight) Glycosylglycerides 20.0 MCT oil (Karlshamns) 49.9 1.0% (w / w) aqueous carrageenan angel (Sigma; Irish Moss, type 1) 30.1 The glycosylglycerides were mixed with the MCT oil to form a homogeneous clear dispersion. The carrageenan gel, which melted at 60 ° C, was slowly added to the oil phase while mixing with vigorous shear. After cooling to room temperature, a highly viscous, milky oil continuous emulsion formed.

Claims (11)

517 678 12 PATENT REQUIREMENTS A lipophilic carrier composition having a continuous lipid phase, comprising a polar, nonionic lipid material in combination with a non-polar lipid, and optionally water, characterized in that the polar lipid material is a glycosylglyceride material, consisting mainly of DGDG but also of MGDG, with the all- The formula R1-o-cuz Rz-o-cn Hzc-o II galactose, cnzo-R, wherein Ra and R2 are independently saturated or unsaturated fatty acid residues having 10-22 carbon atoms, preferably 16-18, and 0-3 double bonds , and RQ is hydrogen or a galactose group; or a mixture thereof. A lipophilic carrier composition according to claim 1, comprising glycosylglycerides, 1-50% by weight, and a non-polar lipid, up to 100% by weight of the total composition. A lipophilic carrier composition according to claim 1, comprising glycosylglycerides and optionally other amphiphiles, 0.5-3.0% by weight, an aqueous solution, 0.1-1.0% by weight, and a non-polar lipid, up to 100% by weight of the total preparation, in the form of inverted vesicles. A lipophilic carrier composition according to claim 1, comprising glycosylglycerides, 1-50% by weight, an aqueous solution 0.1-5.0% by weight and a non-polar lipid, up to 100% by weight of the total composition, in the form of inverted micelles. A lipophilic carrier composition according to claim 1, comprising glycosylglycerides, 1-30% by weight, an aqueous solution, 1-80% by weight, a non-polar lipid, up to 100% by weight of the total composition, in the form of water-in-oil emulsions. 517 678 13 Use of a lipophilic carrier composition according to any one of claims 1-5 as a carrier for an active substance in a pharmaceutical or cosmetic product, food or agricultural product. A pharmaceutical composition comprising a lipophilic carrier composition according to any one of claims 1-5 in combination with a bioactive substance. A pharmaceutical composition according to claim 7, wherein the non-polar lipid is triacylglycerol oil, preferably a medium chain triacylglycerol oil (MCT oil), a palm oil fraction or an active substance. A pharmaceutical composition according to any one of claims 7-8, for oral, enteral, parenteral, rectal, vaginal, topical, ocular, nasal or aural administration. Use of a glycosylglyceride of the general formula R1-O- (E Ez Rz-O- (IIH Hzc-o II galactose * CH2O-Ra wherein R1 and R are independently saturated or unsaturated fatty acid residues having carbon atoms, preferably 16-18, and O-3 double bonds, and R 3 is hydrogen or a galactose group or a mixture thereof, as a polar nonionic lipid to produce a lipophilic carrier composition having a continuous lipid phase comprising a non-polar lipid and a polar nonionic lipid material. Use of a glycosylglyceride of the general formula 517 678 H R 1 -O-cH 2 R 2 -O-cn Hzc-o II galactose cH20-R 3 wherein R 16-18, and 0-3 double bonds, and R 3 is hydrogen or a galactose group; or a mixture thereof, optionally in combination with phospholipids or other amphiphiles, for the preparation of inverted vesicles.