DK171309B1 - Mixture for releasing orally administerable substances - Google Patents

Description

DK 171309 B1 i

The present invention relates to a composition for delivery of orally administered substances. The field of the invention is the biology, and in particular lipid preparations, for the delivery and release of drugs and other substances via the lymphatic system into the systemic circulation.

In terms of drug absorption, they must achieve their goals in a selective and regulative way if their desired pharmacological activities are to be maximized. A 10 method for optimizing the effects of drugs consists in a controlled and sustained administration into the systemic circuit over a long period of time. Orally administered drugs are generally absorbed in the small intestine. Such drugs undergo first-pass treatment through the liver and small intestine; this means that they are converted by the small intestine and by the liver into pharmacologically inactive metabolites and / or excreted in the bile of the liver either as a drug or as active metabolites. As a result, the amount of an orally administered drug which, in effect, enters the systemic circulation may be much less than the amount administered. In order to ensure that effective amounts of such a drug will be included in the circuit and reach the selected target site (s) in the body, larger amounts than those actually required must be administered, and often must be administered in several smaller doses rather than in one. dosage. Orally administered drugs also typically have poor bioavailability. For example, they can are adversely affected by the pH and by the enzymatic activity in the stomach and small intestine, and may be dissolved poorly in the stomach and small intestines.

Numerous attempts have been made to solve these problems and improve the bioavailability of 35 orally administered drugs. The efficacy of certain orally administered drugs has been improved by administering them with a triglyceride or with neutral fat.

Such fats represent a medium which is compatible with lipophilic drugs, ie. that they exhibit low aqueous solubility. Fats also increase the stability of drugs that are unstable in the stomach and in the small intestine. The end products of fat digestion are absorbed by villi in the small intestine mucosa into a lymphatic vessel, the central discharge vessel; absorption occurs within the small intestine region, which results in a limited metabolism of the drug. The absorbed fat is transported through the thoracic duct, the main lymphatic vessel, and then discharged into the blood; it is not introduced into the portal vein blood, which goes to the liver where the first pass metabolism of the drug occurs.

15 The absorption of griseofulvin has been shown to be improved if the drug is administered concomitantly with a high fat meal or in an oil and water emulsion. Crounse, R.G., Journal of Investigative Dermatology, 37: 529 (1961); Carrigan, P.J. and Bates, T. R., Journal 20 of Pharmacological Science, 62: 1476 (1973). If the hormone testosterone undecanoate is administered in a soil nut oil solution, it is more biologically active than if it is administered as an aqueous microcrystalline suspension. Coert, A. J. et al., Acta Endocrinol, 79: 789, 25 (1975); Hirschhauser, C. et al., Acta Endocrinol., 80: 179 (1975). This effect is thought to be due to the absorption of the steroid through the thoracic lymphatic system rather than through the arterial blood; In this way, a first pass through the liver is avoided.

30

Cholesterol, its esters and the triglyceride components (eg fatty acids and monoglycerides) are absorbed through the thoracic lymph. The effects of certain of these compounds alone or in the presence of bile salts on certain orally administered drugs have been studied. For example. led tax per. us of ubidecarenone used in the treatment of hypertension in a mixture containing fatty acids with 12-18 carbon atoms and monoglycerides containing such fatty acids to absorb the ubidecarenone to a greater extent than was found after oral administration of the drug alone (8.3% versus 2.2%). Takl, 5 K. and Takahira, H., U.S. Patent No. 4,325,942. (1982). If the steroid progesterone is administered orally in combination with cholesterol or esters thereof, good long-term biological activity can be achieved. This is thought to be due to the absorption of progesterone via the thoracic lymph and not via the portal vein. Kind, F. A., Proceedings of the 6th International Congress of Pharmacology, 5: 105, (1975).

Yesair has assessed the effect of fatty acids containing 15 12-18 carbon atoms, monoglycerides of these fatty acids and of bile salts on the absorption of estradiol, which is an estrogenic hormone administered orally Yesair, D.W., PCT WO 83/00294 (1983). The molar ratio of the assessed fatty acids: monoglycerides: bile salts ranged from 20 10: 1: 1 to 1: 1: 10 or 1: 10: 1. The preferred ratio is stated to be 2: 1: 2, which corresponds to the micellar composition resulting from the enzymatic digestion of small intestinal triglycerides in the presence of bile salts and calcium ions. When there are 25 bile salts in excess, estradiol incorporated in the 2: 1: 2 mixture can migrate or disperse into a bile salt-enriched micellar solution. This migration or distribution of estradiol occurred prior to absorption of the drug, as evidenced by the fact that the initial concentrations of estradiol in plasma are initially greater than the concentrations in lymph. In addition, approx. 25-50% of the estradiol administered in the mixture was co-absorbed with the lipid components and entered the systemic circulation through the thoracic lymph.

4 DK 171309 B1

The presence of bile salts absorbed in the ileum (and not in the jejunum, as is usually the case with fat) adversely affects the co-absorption of estradiol with fat by increasing the migration of the drug 5 from fat to the bile salt micelle. Phosphatidyl choline was used in an attempt to keep estradiol within the micellar composition in which the fatty acids: the monoglycerides: the bile salts were present in a molar ratio of 2: 1: 2. In the presence of excess bile salts, ca. 60% of the estradiol incorporated in the 2: 1: 2 micellar composition is associated therewith when phosphatidylcholine was not present. Under the same conditions, 70-75% of the estradiol remained in the mixture when phosphatidylcholine was used. However, the addition of phosphatidylcholine for this purpose results in an increased size of the transport system. Size is an important parameter in the absorption of lipid micelles, and this effect of phosphatidylcholine could interfere with the co-absorption of the drug together with the lipids. In addition, excess phosphatidylcholine has been shown to reduce lipid absorption. Ammon, Η. V. et al., Lipids, 14: 395 (1979); Clark, S. B., Gastrointestinal Physiology, 4: E183 (1978). 1 2 3 4 5 6 7 8 9 10 11

Other researchers have also described the effects of 2 the presence of bile salts in lipid formulations used 3 for co-absorption of drugs. Wilson, T. H., 4

In: Intestinal Absorption, Saunders, (1962); Lack, L. and 5 Weiner, I.M., American Journal of Physiology, 240; 313, 6 (1961); Η. V. Ammon et al., Lipids, 14: 395, (1979). For example.

7, there was only a slight difference in the absorption 8 of 5-fluorouracil (5FU) in the stomach or small intestine when 9 5FU was administered alone or in a mixed micelle formulation with mono-olein / sodium taurocholate. The 5FU absorption in the small intestine was greater when the drug was administered in formulation than when administered alone. Streptomycin is poorly absorbed from the intestine. Muranushi and his colleagues report that mixed micelles composed of bile salts, monolayers or unsaturated fatty acids do not improve the absorption of streptomycin from the small intestine; but that they greatly increase the absorption of the large intestine. This increase in the colon was mostly attributed to the change in permeability through the mucosal membrane for mono-olein or for unsaturated fatty acid. In contrast, mixed micelles of bile salts and saturated fatty acids produced only a slight increase in streptomycin absorption even from the colon. Muranushi, N. et al., Journal of Pharmaceutics, 4: 271 (1980). Taniguchi et al. report that mono-olein / tau-rocholate or oleic acid / taurocholate promote the absorption of heparin, which is poorly absorbed when administered alone. Taniguchi, K. et al., International Journal of Pharmaceutics, 4: 219 (1980). The absorption of heparin from the large intestine was twice that of the small intestine. The concentration of heparin in the mixing mold to achieve the improvement in the colon was ca. % of 20 needed in the small intestine.

Sezoshi and Muranishi disclose in U.S. Pat.

No. 4,156,719 is a micelle solution for rectal administration of poorly absorbed water-soluble drugs. Composition 25 consists of fatty acids containing 6-18 carbon atoms and / or mono- or di-glycerides containing the same type of fatty acid; a bile salt or other nonionic surfactant and water. Instead of the fatty acids and mono- or di-glycerides, a lysophosphatidyl-choline moiety can be substituted. Absorption of streptomycin and gentamycin from the rectum and from the large intestine is reported to be comparable in magnitude when the drug is administered into a bile salt micelle: mixed lipid. Similar formulations were not effective in increasing the absorption in the duodenum. Muranushi, S. et al., International Journal of Pharmaceutics, 2: 201 (1979). Absorption of the two drugs via the rectum and colon was markedly greater than it was, with a comparable dose administered in the duodenum, even when the concentration of the mixed lipid micelle administered in the duodenum was 4 times that of the was filed through the other avenues.

5

There is currently a need for a more effective method of improving the absorption of orally administered drugs from the small intestine. A large portion of a drug administered orally never reaches the selected destinations, either because the drug is poorly absorbed or because after it has been absorbed, it is largely removed by the liver. If more effective means were available to increase the absorption of a drug, the treatment would be more effective since only a small amount of the drug was needed to ensure that the desired amount would reach the selected destinations.

With regard to nutrition, the calorie requirements of the individual are primarily a function of the body composition and of the level of physical activity. Medically injured, elderly and physically stressed individuals often have a limited amount of body fat. As a result, the energy needs (calorie requirements) in the main case will be satisfied from 25 external sources.

Physical activities make use of muscles and the energy needs of all muscles including the heart are primarily met as a result of oxidation of fatty acid from fat 30 in the diet or mobilized fatty tissue fat. The adipose tissue fat, as noted, can be quite low and therefore effective absorption of the fat can be an important factor in satisfying the energy needs of the medically impaired, the old and the physically active.

Fat absorption can be adversely affected in many circumstances. For example. For example, in cystic fibrosis, which is a disease state of the exocrine glands, there is a deficiency of pancreatic enzymes, bile salts and hydrogen carbonate ions. Nutrition Reviews, 42: 344 (1984); Ross, C. A., Archives of Diseases of Childhood, 30: 316 (1955): 5 Scow, R. O. E., Journal of Clinical Investigation, 55: 908 (1975). Fat absorption in patients with cystic fibrosis can be quite severely affected and from 30 to 60% of the fat consumed can be malabsorbed. Malabsorption and the resulting steatorrhea are generally not successfully treated by oral administration of pancreatic lipase. In an attempt to regulate the state theory, the patient may appear to be consuming less fat than desirable to achieve good health.

15 Fat absorption may be adversely affected under stressful conditions and the commonly accepted method of dealing with this problem has been to reduce fat consumption. This method can result in both acute and chronic medical problems. These problems 20 can be avoided or at least minimized if a readily absorbable fat source is available.

The present invention relates to a composition consisting of 25 1) non-esterified fatty acids containing 14-18 carbon atoms, 2) monoglycerides which are monoesters of glycerol and fatty acids containing 14-18 carbon atoms, 3) lysophosphatidyl choline wherein the fatty acid component contains 14-18 carbon atoms and 4) a drug.

8 DK 171309 B1

The non-esterified fatty acids and the esterified fatty acid moieties in the monoglycerides and in the phosphatidyl choline may be unsaturated or saturated. In particular, the unsaturated fatty acids may be palmitolic acid, oleic acid, linoleic acid or linolenic acid, which may be present in the mixture individually or in combination. The saturated fatty acids, which also contain 14-18 carbon atoms, can e.g. be myristic acid, palmitic acid or stearic acid.

The non-esterified fatty acids and the monoglycerides are present in the mixture in a molar ratio of between about

2: 1 and approx. 1: 2 (non-esterified fatty acids: monoglycides). The lysophosphatidyl choline constitutes from ca. 1.0 mole% to approx. 30.0 mole% of the total mixture.

15

If the non-esterified fatty acids in the mixture are saturated, sufficiently large amounts of divalent cations (about half of the molar amount of the fatty acids), such as calcium ions, are added to form non-esterified fatty acid salts.

The mixture of the present invention is intended to promote the uptake of the mixed lipid colloid into the small intestine mucosa, the subsequent synthesis to 25 chylomicrons, the translocation of the chylomicrons to the thoracic lymph, and the transport to the systemic circulation (i.e., blood).

The composition of the present invention will promote rapid and quantitative absorption of lipids in the small intestine and transport of the lipids via the lymphatic vessels due to several characteristics. First, the molar ratios described for the fatty acids and monoglycerides are optimal for their absorption in the jejunum. Second, it has been shown that the unsaturated fatty acids or the saturated fatty acid calcium salts included in the mixture are maximally absorbed and preferably transported via the thoracic lymph (and not transported via the arterial blood). Third, the mixture also contains lysophosphatidyl choline which promotes the translocation of the reconstituted lipids as chylomicrons into the thorax lymph.

The composition of the present invention can serve as a carrier for increased uptake and bioavailability of drugs. Drugs are widely defined here as any chemical agent or chemicals that exert an influence on life processes. Examples of substances which may be incorporated into the compound of the present invention are drugs administered for diagnostic, therapeutic or preventive purposes; 15 lipophilic prodrugs; bioactive peptides; certain nutrients such as fat-soluble vitamins and other xeno-biotics. This increase or advance occurs because the drug incorporated in the composition of the invention is absorbed together with lipids and as a result enters the systemic circulation through the lymphatic vessels. As a result, it is absorbed faster and more completely than would otherwise be the case. Because a first passage through the liver is avoided, more of the absorbed dose enters the bloodstream and is available to reach the target areas than would be available if the lipid formulation was not used.

The mixture of the present invention may also serve as a highly concentrated source of easily absorbable fat, such as e.g. can be used by such individuals who need a calorically heavy dietary component. The mixture of the present invention, when used in this way, consists of 1), 2) and 3) as described above, and it does not include a drug.

10 DK 171309 B1

The mixture according to the present invention also provides a stable, mixed 11pld collold which will protect the drugs from e.g. enzymatic and chemical degradation in the stomach and the upper part of the intestine. In addition, the built-in stability of the lipid components will make the mixtures stable over a longer period of time and thus serve as stable transport media for the substances incorporated in the formulation.

10 In the drawing, FIG. 1 in a graph depicting the total vitamin A-derived radioactivity equivalents over time in plasma of rats given vitamin H in a mixture and in plasma from rats given 3 vitamin H in corn oil .

15 In FIG. 2 is a graphical representation of the mean plasma concentration-time profiles of N- (4-hydroxyphenyl) -trans-retinamide (HPR) in dogs after oral HPR administration.

20

The composition of the present invention, which consists of unesterified fatty acids, monoglycerides of such fatty acids, lysophosphatidyl choline containing such fatty acids as its fatty acid moiety, and a drug, is based on the absorption and transport characteristics of the fatty acids and on the contribution of lysophosphatidyl. choline to solubilize the drug in the lipid mixture and to translocate the absorbed fat into the lymph (rather than into the portal vein).

30

Hydrolysis of triglycerides leads to a molar ratio of fatty acid to monoglyceride of 2: 1. Upon subsequent absorption and re-synthesization of the intestinal mucous layer, fatty acids are mixed from the food and lose their identity; new triglycerides are formed which are partly characteristic of the nature of the organism concerned.

11 DK 171309 B1

Two components of the bile are fatty acids and phosphatidylcholine. Enzymatic hydrolysis of phosphatidylcholine leads to fatty acids; complete hydrolysis leads to 2 molecules of fatty acid and partial hydrolysis leads to 1 molecule of 5 fatty acid and lysophosphatidyl choline. As a result, a mixture containing an excess of monoglyceride (e.g., fatty acid: monoglyceride ratio of 1: 2) may be in equilibrium with such fatty acids derived from the bile, at approximately the optimal ratio of 2: 1 for the 10 improved absorption of mixed lipid colloids from the upper intestine. A ratio of fatty acid: mono-glycerides not in this range (ie 1: 2 to 2: 1) cannot promote lipid absorption.

It can also adversely affect not only the lipid absorption, but also the co-absorption of drugs administered with the lipids.

A less effective way of adjusting to the excess fatty acid is the addition of glycerol instead of monoglycerides. If the incoming lipids have not been absorbed within the jejunum area of the intestine, they continue in the small intestine to the colon. The drugs that move with the lipids can be absorbed and transported via the portal vein for excretion at first passage 25 through the liver. Once in the large intestine, the incoming lipids of the intestinal microflora are used. These microorganisms can also metabolize the drug that moves together with the incoming lipids to form toxicologically active species. Goldman, P. In: 30 The Molecular and Cellular Approaches to Understanding the Mechamisms of Toxicity, A. H. Tashjian (ed.) 164, (1983).

It has been shown that absorption of saturated fatty acids is inversely proportional to the number of carbon atoms in the fatty acid. For example. the absorption of decanoic acid (10: 0, which indicates the chain length and the extent of unsaturation) is almost quantitative. For lauric acid (12: 0) it is more than 95%; for myristic acid (14: 0) it is 80-90%; for palmitic acid (16: 0) it is 65-70% and for stearic acid (18: 0) it is 30-45%.

5

The transport of absorbed fatty acid via the lymph (and not in the portal vein) varies widely. That is, it has been shown that the much larger percentage of absorbed unsaturated fatty acids is carried in the lymph than in the case of saturated fatty acids. It has been shown that approx. 85% of the unsaturated fatty acids are carried in the lymph. Miura, S. et al., Keio Journal of Medicine, 28: 121 (1979). In contrast, 5-20% of absorbed decanoic acid (10: 0) and 15-55% of absorbed lauric acid (12: 0) are transported into the lymph, although 95% of 15 will be absorbed a given amount of each. The remaining portion is absorbed into the portal vein blood. As noted above, the percent absorption of saturated fatty acids is inversely proportional to the chain length. That is, myristic acid (14: 0) is absorbed to a greater extent than 20 palmitic acid (16: 0), which in turn is absorbed to a greater extent than stearic acid (18: 0). The amount of these fatty acids absorbed into the lymph is also inversely proportional to the chain length: 68-80% for myri stic acid; 85% for palmitic acid and stearic acid. In addition, 25, it has been shown that saturated fatty acids (e.g. linoleic acid 18: 2) are absorbed more quickly into the lymph and to a greater extent than saturated fatty acid. Taniquchi, K., International Journal of Pharmaceutics, 4: 219 (1980). 1 2 3 4 5 6

If saturated fatty acids are included in the composition of the present invention, they are included in form 3 of calcium salts or salts of another cation. This is the case 4 because the enzymatic hydrolysis of triglycerides which release saturated fatty acids promotes the formation of the 6 calcium soaps thereof. Thank you, Y.A. and Grigor, M. R., Biochemistry Biophysica Acta, 531: 257 (1978).

13 DK 171309 B1

It has been shown that a transfer of absorbed fat to the lymph requires lysophosphatidyl cholene. Both lysophosphatidyl-choline and phosphatidyl-choline Contain phosphoryl-choline bound to C-1 in glycerol; but they differ 5 by the number of fatty acids they contain. Lyso-phosphatidylcholine has a single fatty acid component located at either C-1 or C-2 in the molecule; phosphatidylcholine has two fatty acid moieties (one at C-1 and one at C-2). The rate, but not the extent, of the transmission 10 of absorbed fat is apparently associated with the fatty acid portion of lysophosphatidyl choline. For example. oleoyl-lysophosphatidyl-choline (derived from di-oleoyl-phos-phatidyl-choline) results in a 100% increase in triglyceride and phospholipid transported through the lymphatic vessels when compared with the effects of a lysophosphatidyl-choline, which is derived from a phosphatidyl-choline composed mainly of saturated fatty acids (e.g. palmitic acid (C16: 0) and stearic acid (C18: 0)). There is, in the absence of luminal phosphatidyl choline, to lead to a lysophosphatidyl choline, an increased accumulation of mucosal triglyceride and apparently increased portal vein transport of the absorbed fatty acids. Tso, P et al., Gastroenterology, 80:60 (1981); 0'Doherty, P.J.A. et al., Lipids, 8: 249 (1973). If the load of fat is increased 5 times, a defect is seen in the lymph transport of absorbed fat. This effect can be counteracted by the addition of a source of lysophosphatidyl choline. Tso, P. et al., Gastroenterology, 73: 1362 (1977). This effect of lysophosphatidylcholine observed in vivo is also found in vitro. Rodgers, J. B., and O * Connor, P. J., Biochemistry Biophysica Acta, 409: 192 (1975). Lysophosphatidyl choline has been shown to be rapidly absorbed intact. Rodgers, J. B. et al., Digestive Diseases, 20: 208 (1975); Nilsson, A., Biochimica Biophysica Acta, 137: 240, (1976).

Lymph phospholipids have also been found to contain substantially unsaturated fatty acids: oleic acid (18: 1) 50-63%; linoleic acid (18: 2) 17-20% and arachidonic acid (20: 4) 2-4%. Incorporation of an unsaturated lysophosphatidyl choline into the composition of the present invention will facilitate the transfer of the absorbed lipids and the drugs or other substances thus absorbed. In addition, the lysophosphatidyl choline plays a role in the solubilization of certain drugs (for example, the presence thereof increases the solubility of the drugs in the mixture).

10

The mixture of the present invention consists of non-esterified fatty acids containing 14-18 carbon atoms; monoglycerides which are monoesters of glycerol and fatty acids containing 14-18 carbon atoms; lysophosphatidyl choline, in which the fatty acid portion also has 14-18 carbon atoms as well as a drug. The non-esterified fatty acids and fatty acids in the monoglycerides and in lysophosphatidylcholine may be unsaturated or saturated, and they will preferably be unsaturated. Examples of unsaturated 20 fatty acids which may be used in the composition of the present invention are: palmitolic acid CH16H30 ° 2 16: 1 oleic acid ^ H18H34 ° 2 18: 1 25 linoleic acid CH ^ gH ^ C ^ 18: 2 linolenic acid CH18H30 ° 2 18: 3

Examples of saturated fatty acids which can be used in the composition of the invention are: 30 myristic acid CH14H28 ° 2 14: 0 palmitic acid CH16H32 ° 2 16: 0 stearic acid CH18H36 ° 2 18: 0 35 The non-esterified fatty acids and monoglycerides are present in amounts which leads to a molar ratio of approx. 2: 1 to 1: 2 (non-esterified fatty acid: monoglycide).

In addition, the mixture contains lysophosphatidyl choline, whose fatty acid moiety contains 14-18 carbon atoms and is preferably unsaturated. The fatty acid component of the lysophosphatidyl choline is preferably one of the above. The amount of lysophosphatidyl choline in the mixture is determined by the amount needed to increase the solubilization of a drug to be administered in the mixture and the amount needed for its transmission. Lysophosphatidyl choline generally amounts to from ca. 1.0 mole% to approx. 30.0 mole% of the total mixture. The fatty acids included in the composition of the present invention, whether these are non-esterified fatty acids or are constituents of monoglycerides or lysophosphatidylcholine, may all be similar or a number of different fatty acids may be included.

The composition of the present invention may also contain a drug which may be any chemical or chemical which influences living processes. They include, but are not limited to, drugs administered for diagnostic, therapeutic or preventive purposes; lipophilic prodrugs; certain nutrients such as fat-soluble vitamins and other xenobiotics.

The mixture of the present invention is prepared according to the method described in the following. The incoming lipid components are weighed and mixed with or without the use of heat to achieve fluid homogeneity. The drug is added and dissolved with or without the use of heat in the lipid mixture. A uniform state is indicated by the absence of solids at the appropriate temperature at which the mixture is to be liquid and by the absence of blurring. A blurring effect will be more evident at greater concentrations of the drug in the lipid mixture. The formulation is stable to several cycles of freezing and thawing? the appearance of solids or of blurring may indicate instability of the formulation.

Another method of preparing the formulation involves dissolving the incoming lipids and drug in a solvent or in a mixture of solvents and mixing to achieve homogeneity. The solvents are removed under vacuum or other suitable methods. The criterion for a suitable formulation will be the same as stated above.

The mixture of the present invention is illustrated by the following examples.

EXAMPLE I

Preparation of the mixture and comparison with corn oil as a means of transport

Lipid mixture 3 25 Tritium-labeled (H) vitamin A (7 mg or 0.25 mmol; 30 curie per mole; specific activity 1.26 mC / mmol; Hoffman LaRoche) was incorporated into a lipid formulation of the following composition: 56 mg of oleic acid (NuCheck Pr ep,

Inc.); 36 mg mono-olein (NuCheck Prep, Inc.) and 6 mg 30 lysophosphatidyl choline derived from soy lecithin (A. E. Staley Mfg. Co.). This corresponds to an approximate molar ratio of oleic acid: monoolein: lysophosphatidylcholine of 2: 1: 0.1. (The approximate molar percentages of the constituent components were 64.5: 32.3: 3.2, respectively).

The mixture was administered to rats of the species Sprague-Daw-ley (weight 210-230 g) by forced feeding.

17 DK 171309 B1

May sun preparations

Tritium-labeled vitamin A (7 mg or 0.25 mmol) was administered to Sprague-Dawley rats (as described above) in 5 99 mg of food grade refined corn oil (A.E.

Staley Mfg. Co.) ·

Experimental Procedure and Results 10 The concentration of vitamin A equivalents in plasma was measured by taking 0.05 ml double blood samples from the orbital sinus 0.5, 1, 2, 3, 4, 6, 8 and 24 hours after administration. The blood was collected in hematocrit tubes and centrifuged. The plasma was transferred to scintillation vials and weighed. Radioactivity was determined by liquid scintillation counting.

The results indicated a slow absorption of vitamin A over an 8 hour period (Fig. 1). A calculation of the amount absorbed as a function of time is shown in Table 1. Approx. 73% of vitamin A in a mixture was absorbed within 2 hours of administration; About 50% of vitamin A administered in corn oil was absorbed within the first 2 hours. At 3 hours 25 post-administration, more than 90% of vitamin A in the lipid mixture and 77% of vitamin A in the corn oil had been absorbed.

At the end of the 8-hour study period, the absorption of vitamin A in both the mixture and in corn oil was complete. The plasma concentration of tritium-labeled vitamin A equivalents in rats in the two groups was comparable. The mixture increased the absorption of the vitamin A incorporated into it and also caused the vitamin to be absorbed more quickly.

18 DK 171309 B1

The areas under the concentration-time curve for the 8-hour period were also equivalent, indicating that the absorption of vitamin A in the mixture was quantitative. By a similar comparison, approx. 73% of vitamin A in the mixture was absorbed within 2 hours (Table 1) and the absorption was greater than 90% at 3 hours compared to 52 and 77% for vitamin A administered in corn oil, respectively. Thus, it is clear that the mixture will cause both the elevated and the earlier absorption of incorporated drugs and therefore increase their efficacy.

TABLE 1: Proportional absorption of vitamin A administered in the mixture or in corn oil to rats by oral incubation.

Percent absorption

Lipid Ratio 20 Hours Blend Corn Oil Blend / Oil 0 - 0.5 5.4 4.0 1.35 0. 5- 1 27.3 20.0 1.37 1- 2 72.6 52.4 1.39 2 3 92.0 77.2 1.19 3-4 97.3 91.8 1.06 4-6 100.0 99.5 1.01 6-8 100.0 100.0 1.00 EXAMPLE II 30

Assessing the capacity and stability of a mixture as a means of transporting vitamin A

A mixture comprising oleic acid, monoglyceride and lysophosphatidylcholine, as described in Example 1, was used to characterize the mixture's capacity for vitamin A (also as described above). To the mixture, DK 171309 B1 19 was added to vitamin A 1 portions as follows: 1. Addition: 0.277 mmol 2. Addition: 0.241 mmol 5. Addition: 0.246 mmol 4. Addition: 0.262 mmol and 5. Addition: 0.285 mmol.

The 5th addition resulted in the addition of 1.34 mmol of 10 or 3.84 mg of vitamin A to ca. 1 g of the mixture.

The millimole ratio of the mixture was approx. 2: 1: 0.1 (oleic acid: monoolein: lysophosphatidylcholine).

After each addition, the approximate time needed to completely dissolve the added vitamin A, as well as the extent of visible blur, was noted.

The time needed to obtain a minimal blur was generally between 2 minutes and 60 minutes and was inversely proportional to the amount of vitamin A.

After each addition and obtaining a minimal blur, the solution was placed in ice water to assess its stability. The relative viscosity of the cold solution was also noted. The results obtained are summarized in Table 2 below.

25

A clear solution without blurring was observed within 1 hour after the 5th addition. The same solution was frozen for 1 week and thawed. A clear solution was observed at that time. This cycle between freezing and thawing was repeated for 4 hours over a period of 8 weeks with comparable results. Thus, it is clear that this lipid formulation is stable over longer storage periods and over changes in temperature.

35 DK 171309 Pg 20

Table 2: Vitamin A mixture capacity

Condition to obtain

Vitamin A_clear solution 5

Added Total (mmol) (mmol) Time Temp. Blur Viscosity 0.277 0.277 1 min. 20 ° C no normal 0.241 0.518 1 min. 20 ° C minimum normal 10 0.276 0.794 5 min. 37 ° C medium thicker 0.262 1.056 10 min. 37 ° C medium viscous 0.285 1.341 10 min. 46 ° C comprising very sealing viscous

EXAMPLE III

Assessment of the Solubility of N- (4-Hydroxyphenyl) -trans-Retinamide (HPR) in Lipid Formulations 20 Lipid Formulations

N- (4-hydroxyphenyl) -trans-retinamide (HPR) (McNeil Pharmaceutical) was used to characterize its solubility in several formulations of the composition of the present invention. The composition of the mixture varied with respect to the fatty acid constituents among the 3 basic components: fatty acid (NuCheck Prep Inc.), monoglyceride (NuCheck Prep Inc.) and lysophosphatidylcholine (Avanti Polar Lipids). The formulations used are listed in the later Table 3.

In each of the formulations, the lipid components were first mixed together in a sealed container under nitrogen. Then solid HPR was added in small portions. The container 35 was again sealed under nitrogen after each addition and heated and treated with ultrasound to form a solution of HPR, or until it became clear that the solid HPR would remain in suspension. Such formulations which resulted in dissolution of HPR were cooled in the refrigerator or frozen, and continued solubilization of HPR was noted. In all cases, once solubilized, HPR, 5 did not precipitate out of solution.

The results obtained from the observations for the formulations in question are shown in subsequent Table 3. In Formulations II and III, resolution of HPR was noted when the molar ratio of lysophosphatidylcholine (LysoPC) to HPR (LysoPC: HPR) was observed. greater than 1; a suspension of HPR was observed when the molar ratio of lysophosphatidylcholine to HPR was less than 1.

When comparing Formulations III and IV, which contained approximately the same amount of HPR, the ratio LysoPC: HPR was less than 1 in Formulation III when a suspension of HPR was observed and greater than 1 in Formulation IV when a solution was observed. . Comparison of Formulas V and VI showed that the solution of HPR was clearly visible at molar ratio of 1 (LysoPC: HPR) where lysophosphatidylcholine containing either myristyl fatty acid (14: 1) or palmityl fatty acid (16: 1) ). In Formulation V, the solubilization of the third sequential addition of HPR appears to be more difficult than the first two additions. This is a further indication of the importance of having a molar ratio of LysoPC: HPR greater than 1. 1 2 3 4 5 6

Formulation VII represents a formulation of HPR as 2 contained all components of the aforementioned for 3 formulations. As described in Example 4 below, this formulation was administered to dogs.

5

Briefly, the ratio of 18: 1 LysoPC: HPR was 0.69; ratio 6 of 14: 0 LysoPC: HPR was 0.33; and the ratio of 16: 0 LysoPC: HPR was also 0.33. The molar ratio of lysophosphatidylcholine: HPR in Formulation VII was 1.31: 1.

22 DK 171309 B1

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EXAMPLE IV

Biological availability of N- (4-hydroxyphenyl) trans-retinamide (HPR) administered in a formulation 5

This study was designed to compare the bioavailability of HPR from Lipid Formulation VII (see Example III and Table 3), administered to fasting beagle dogs (treatment E 2) and dogs under 10 feeding (treatment E 2), with the bioavailability of HPR administered to beagle dogs in a reference formulation (treatment A) during feeding mode. The reference formulation was an oil-based mixture containing a surfactant.

15 In these studies 4 dogs of the species beagle were used; all dogs received HPR, with at least 1 week between treatments. The dose of HPR for treatment A and E ^ was 200 mg; it was administered in suspension in treatment A and in solution in treatment, and in every 20 cases it was administered in the form of two 100 mg capsules. In treatment E2, 100 mg of HPR was administered in solution as a 100 mg capsule. All data for treatment E2 have been presented with a normalization to a dose of 200 mg.

25

The dogs received food and water ad libitum during treatment A and treatment E2. They were fasted for food for 12 hours before receiving treatment E1; this fasting continued until 2 hours after dosing.

30

Plasma samples for HPR were analyzed using the high pressure liquid chromatography method. Plasma concentration of HPR in this study is shown in summary in Table 4 and in FIG. 2nd

The mean HPR peak concentrations were 1660, 4238 and 8174 ng / ml for treatment A, E1 and E2, respectively (Table 4). The mean area under the curve, AUC (0-80 hours), showed values of 22666, 62590 and 95935 ng x hour x ml for treatment A, Ej and E2, respectively (Table 5). It is readily apparent that the bioavailability of HPR Submitted 1 Formulation VII (treatment Ej and E2) was greater than the bioavailability of HPR filed in the reference formulation (treatment A). A comparison of the results obtained when administering Formulation VII to the animals in feeding state (E 1), with results obtained when the formulation was given to fasted animals (E 1) indicates that additional lipids (from the fed food) result in a further increase in AUC (1.5-fold increase between treatments E2 and E1). The unsaturated lysophosphatidyl choline is particularly useful in the transfer of absorbed fat to the lymph. Since these unsaturated lysophosphatidyl choline may have been combined with HPR, there was insufficient 18: 1 LysoPC to both solubilize HPR and transfer the absorbed lipids and HPR to the 20 lymph. As a result, such drugs which interact with lysophosphatidyl choline in the lipid formulation apparently require a higher molar content of lysophosphatidyl choline with respect to the fatty acids and monoglycerides.

25 1 35 DK 171309 B1 26

Table 4: Summary of plasma concentration-time data for HPR in 4 dogs after oral administration of HPR in Formulation VII

5 Average HPR concentration in plasma ng / ml

Formulation Reference A Lipid E ^ Lipid-E3 state Lined Fasting Lined 10

Time (hours) Plasma HPR concentration (ng / ml) 0 0.0 24.0 121.9 0.5 219.9 238.9 345.5 1 769.7 1093.3 2388.3 15 2 1430.6 3124 7 7259.8 3 1490.5 3866.0 8032.5 4 1325.2 4078.8 6841.1 5 1105.2 3599.1 5411.0 6 914.3 3021.3 4386.7 20 7 797.3 2470.7 3402.7 24 187.4 557.5 852.6 30 198.0 426.5 646.1 48 93.9 253.3 449.3 54 79.4 193.6 333.9 25 72 69, 7 149.0 276.0 80 47.8 114.5 210.3 sk

Corrected to baseline dose of 200 mg. 1 35 27 DK 171309 B1

Table 5: Area under the plasma concentration-time curve [AUC (0-80 hours)] for HPR in 4 dogs after oral administration of HPR in Formulation VII

5 HPR-AUC (mg H / ml)

Formulation Reference A Lipid Lipid-E2 * Condition Lined Fasting Lined 10 Dog no.

1 28609 50044 107106 2 15964 85898 127589 3 21190 78476 56274 4 24900 36941 92721 15 --------------------------------- ------------------

Middle 22666 62590 95935

Standard Deviation 5398 22842 30054 20 Median 23045 64260 99939 G. Mean 22156 59241 91905

25 As a% of A

Medium 276 423

Median 279 434 G average 267 415 30 a Corrected to a baseline dose of 200 mg 35

Claims (11)

28 DK 171309 B1 Patent claims: A composition for delivery of orally administrable substances, characterized in that it consists of: a) at least one non-esterified C14-Clg fatty acid, b) at least one monoglyceride which is a glycerol monoester and a C14 ~ Clg fatty acid, c) lysophosphatidyl choline wherein the fatty acid moiety contains 14-18 carbon atoms and 15 d) a drug, the ratio of components a) and b) being from 1: 2 to 2: 1. A composition according to claim 1 for increasing the absorption of a drug and for increasing the transport of non-esterified fatty acids and monoglycerides in an individual's lymphatic vessels, characterized in that it consists of: a) non-esterified C 14 ~ C g fatty acids (B) monoglycerides which are monoesters of glycerol and a C 14 -C 18 fatty acid; to solubilize the drug 35 in the mixture. 29 DK 171309 B1 A composition according to claim 1, characterized in that it consists of: a) at least one non-esterified C ^-C ^g fatty acid and at least one monoglyceride which is a glyo-rol monoester and a C ^-C ^g fatty acid, which fatty acid and which monoglyceride together constitute 70.0-99.0 mol% of the mixture and is present in the mixture in a molar ratio of between 1: 2 and 2: 1, 10 b) lysophosphatidyl choline wherein the fatty acid moiety contains 14- 18 carbon atoms, which phosphatidyl choline constitutes 1.0-30.0 mol% of the mixture and 15 c) a drug. A composition according to claim 3, characterized in that the molar ratio of fatty acid to monoglyceride is about 2: 1 and that the lysophosphatidyl choline is 3.0 to 13.0 mol%. Lipid-containing composition according to claim 3, characterized in that it contains non-esterified saturated C ^-CCg fatty acids and a sufficient amount of calcium ions to form a salt with the non-esterified saturated fatty acids. Composition according to claim 1, characterized in that it consists of: a) at least one non-esterified fatty acid selected from palmitolic acid, oleic acid, linoleic acid, linolenic acid and saturated c14 'C1g fatty acids, b) at least one monoglyceride which is a monoester of glycerol and a fatty acid selected from palmitic acid, oleic acid, linoleic acid, linolenic acid and saturated C 1 -C 2 g fatty acids; d) a drug, the fatty acid and the monoglyceride together constituting 70.0-99.0 mole percent of the mixture and present in a ratio of fatty acid to monoglyceride between 1: 2 and 2: 1 and the in-lysophosphatidyl choline constitutes from 1.0 to 30.0 mole percent of the mixture. 15 A composition according to claim 1, characterized in that it consists of: a) non-esterified C C-C ^g fatty acids, b) monoglycerides which are monoesters of glycerol and a C) lysophosphatidyl choline, wherein the fatty acid moiety contains 14-18 carbon atoms, and d) a drug, the unesterified fatty acids and the monoglyceride together amounting to 87.0-97.0 mole. % of the mixture and is present in a molar ratio of fatty acid to monoglyceride of about 2: 1, with the lysophosphatidyl choline accounting for 3.0-10 13.0 mol% of the mixture. A composition according to claim 1, characterized in that it consists of: a) at least one non-esterified C ^-C fatty acid, c) lysophosphatidyl choline, wherein the fatty acid moiety contains 14-18 carbon atoms, and d) a drug. Lipid composition according to claim 7, characterized in that the incoming drug is a fat-soluble vitamin. 15 Mixture according to claim 7, characterized in that the non-esterified fatty acid, the fatty acid in the monoglyceride and the fatty acid in the lysophosphatidyl choline are each selected from palmitic acid, oleic acid, linoleic acid, linoleic acid, myristic acid, palmitic acid and stearic acid. A composition according to claim 7, characterized in that the non-esterified fatty acid is oleic acid, the monoglyceride is monoolein or monopalmitin, and the lysophosphatidyl choline is selected from lysophosphatidyl choline derived from soya lecithin, myristyl lysophosphatidyl choline. lysophosphatidyl-choline and oleyl-phosphatidyl-choline. A composition according to claim 1, characterized in that it consists of: a) oleic acid and monoolein in a molar ratio of about 2: 1, 35 b) approx. 3.0 mole% lysophosphatidyl choline; and c) vitamin A.