DE2915028A1 - ROYALTY vehiculum TO SUPPLY AND RELEASE OF A xenobiotic WITHIN A SAEUGETIERWIRTS, PROCESS FOR PRODUCING THE ROYALTY VEHICULUMS AND ITS USE IN A PROCESS FOR PREDESTINATION AND CONTROL OF pharmacodynamic UNDER WHICH AN INSIDE A xenobiotic SAEUGETIERWIRTS DELIVERED OR WILL BE RELEASED - Google Patents

Description

B eschre i bung

The invention relates to the delivery and release of xenobiotics within a mammalian host in particular vehicles that release or release xenobiotics in the form of circulating microreservoirs, a method for the production of the microreservoirs and a process for the delivery or release of xenobiotics a mammalian host that allows the pharmacodynamic properties of the delivered and released xenobiotics can be predetermined and controlled.

In many different situations and under many different In some circumstances it is desirable to introduce pharmacologically active agents into a mammalian host that are effective for are alien to the host organism; these agents become hereinafter referred to as "xenobiotics". These xenobiotics include, but are not limited to, drugs or drugs, diagnostic agents, blood substitutes, endogenous biological compounds or mixtures, Hormones, immunological adjuvants and the like.

When administering any xenobiotic, a certain degree of specificity must be achieved and the specificity requires the xenobiotic to achieve its goal in a selective and controllable manner. The lack of Specificity in the use of many xenobiotics can therefore make them to a considerable extent, if not practical fully, of their potential effectiveness in the

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Depriving of achieving the results sought after in their use. For example, a chemotherapeutic one is missing Medicines that do not stay in blood plasma for a long enough time can be retained so that a significant amount of the drug reaches the target tissue, or an orally administered medicine intended for the bloodstream but not the gastrointestinal tract can happen, the degree of specificity, the Θ3 could make highly effective. In the absence of the desired specificity, a xenobiotic can have practically no or have only a limited degree of the desired pharmacodynamic properties that are desired, to realize its full effectiveness. These pharmacodynamic properties include, for example Plasma kinetics, the distribution within the tissue, the toxicity, the levels of the therapeutic drugs in vivo, the solubility of xenobiotics, which are normally incompatible with other pharmaceutical formulations and the metabolic activation of xenobiotics.

It was previously believed that it would be desirable to determine the specificity or pharmacodynamic properties of many of the Xenobiotics that have been found to have desirable properties to modify and control (steer). An attempt to control the specificity of drugs involves the use of implant devices, which, usually with the help of a surgical Procedure in or near the organ to which the drug is delivered (released) should be. As a rule, these implant devices consist of a covalent matrix material, which is the drug to be delivered or released

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contains. These matrix materials can be water-soluble (such as Carboxymethyl cellulose or polyvinyl alcohol), quenchable in water (such as hydrogels or gelatin), hydrolytic Polymers (such as polylactic acids, polyglycolic acids or polyamino acids) or non-hydrolytic polymers (such as organopolysiloxane rubber) be. Although by these implant devices the rate or speed with who controls the drug they contain by diffusion or hydrolysis can thus generally only be a slight, if any, change in the pharmacodynamic Properties of the released drug can be achieved. One of the more recent attempts at the pharmacodynamic To modify and control properties of xenobiotics is the use of a carrier that the xenobiotics the center of action within the Host to which the xenobiotics are to be administered can deliver and release in a controllable manner. It the use of liposomes as carriers for this purpose has already been proposed (cf. G. Gregoriadis, "The Carrier Potential of Liposomes in Biology and Medicines ", in" The New England Journal of Medicine ", Volume 295, No. 13, pp. 704-710 (September 23, 1976), and Mr. 14, p. 765 to 769 (September 30, 1976)). These well-known liposomes consist of phospholipids and especially of phosphatidylcholine in combination with other lipids, such as Cholesterol, dicetyl phosphate, stearylamine and others Phospholipids, such as phosphatidylserine, with which the phosphatidylcholine is easily miscible. The liposomes are through characterized in that they are miscible with the carrier components, a fact that means that between little or no of these components

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Phase separation can exist. In addition, these beams lack a high degree of stability, primarily due to the oxidation of the phospholipid components and / or their thermal instability, especially if they are up to achieve a small particle size, for example about 250 Å, treated with ultrasound (sonicated). In addition, those liposomes that carry a negative charge aggregate due to the presence of Dicetyl phosphate, phosphatidylserine or other negative charged components in the presence of divalent cations. If liposomes are injected intravenously, concentrate they eventually prefer to live in the liver and spleen, due at least in part to their large particle size is due. Although the known liposome carriers are compatible with a number of drugs and are biocompatible within the mammalian system, their inherent instability becomes their considerable Tendency to aggregate and their relatively large particle size their ability to be acceptable Drug delivery system to function.

Because of the ever increasing role that xenobiotics have inherited in treating cancer, diabetes, arthritis Metabolic disorders, metal storage disorders and the like, in the prevention of diseases such as anaplasmosis (anaplasmosis) and viral infections, and in the Investigating biological mechanisms at play, the need for improved carriers is constantly increasing. Of the The desire to have such improved delivery systems is therefore evident.

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The primary object of the present invention is to provide improved xenobiotic delivery vehicles in the form of To create micro-reservoirs within the host— systems can circulate. Another object of the invention is to provide xenobiotic delivery vehicles Find those with a wide variety of xenobiotics including hydrophobic, hydrophilic, or a combination of hydrophobic and hydrophilic compounds or mixtures are compatible, are non-toxic and with the Are biologically compatible with the host system. Another aim of the invention is to provide X.enobiotikcT - Zuführungsbzw. -Delivery vehicles to be found over long periods of storage as well as persistent over extended periods of use within the host system various routes of administration including oral, intravenous, intramuscular, intraperitoneal, subcutaneous, topical, and inhalation administration. Another object of the invention is in finding X.enobiotics - delivery vehicles that are capable of acting on predictable and beneficial way the pharaakod; / namic properties to change and control the supplied xenobiotic and released within the host system. The pharmacodynamic properties that are advantageously modified and controlled in this way include the plasma kinetics, the distribution within the tissue, the toxicity, the oral absorption, the chemotherapeutic Effectiveness, metabolism and the like.

Another main object of the invention is to provide a method for making xenobiotic delivery or delivery vehicles in the form of microreservoirs that

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can circulate within a mammalian host, develop, with the help of which it is possible to deliver the xenobiotic to a given place by placing a given causes beneficial change in the pharmacodynamic properties of the xenobiotic.

Finally, a third main object of the invention is to provide a method for delivering and releasing a pharmaceutically effective amount of a xenobiotic within of a mammalian host, with the help of which it is possible to establish predictable control over the Exercise delivery or delivery center so as to improve the effectiveness of the xenobiotic.

Other objects, features and advantages of the invention will appear from the following description.

In one aspect, the present invention relates to a A delivery or delivery vehicle that is biologically compatible with a mammalian host (always here for the sake of simplicity referred to as a "delivery vehicle") to deliver a xenobiotic to and within the host organism to release its pharmacodynamic properties by using the vehicle in an advantageous manner Manner changed or changed v / ground, which is characterized in that the supply or delivery vehicle in "In the form of microreservoirs which contain the xenobiotic, the microreservoirs contain or exist of a phospholipid component and a lipid component which is immiscible with the phospholipid and which is contained in a physiologically compatible liquid is stable and essentially immiscible with it.

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In another aspect, "relates to the present invention a method for making a xenobiotic delivery or delivery vehicle for the introduction or delivery of a xenobiotic within a mammalian host, whose pharmacodynamic properties are modified and controlled in a predictable way, that is is characterized in that one microreservoirs from one Phospholipid component and a lipid component which is immiscible with the phospholipid and which is in one physiologically compatible liquid is stable and therefore practically immiscible, and the to be supplied or to be released xenobiotic incorporated into the microreservoirs.

In another aspect, the present invention relates to a method of delivery (delivery) and release a xenobiotic within a mammalian host characterized by being in the mammalian host introduces a pharmaceutically effective amount of a xenobiotic contained in microreservoirs provided by a physiologically compatible liquid and made up of a phospholipid component and a lipid constituents which are immiscible with the phospholipid and which are in the physiologically compatible Liquid is stable and therefore essentially immiscible.

According to a further aspect, the present invention finally relates to a method for prediction and Control of the pharmacodynamic properties or conditions under which a xenobiotic within a Mammal host is released, which is thus identified

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What draws is that you get the xenobiotic inside the host from the circulating microreservoirs released from one phospholipid component and one with the phospholipid Immiscible lipid constituents exist that are stable and in a physiologically compatible liquid so it is essentially immiscible.

The present invention therefore comprises several stages and the relationship of one or more of these stages to one another, the composition or preparation and! which have the characteristics and properties, as well as the relationships of the constituents among themselves, which are in the will be explained in more detail below. To better understand the nature and objectives of the In the following, the invention is described in more detail with reference to the accompanying drawings. Included demonstrate:

Figure 1 is a process flow diagram illustrating a process for producing the xenobiotic delivery vehicles of the invention explained;

Fig. 2 is a partial process flow diagram showing a! modification of the method according to FIG. 1 explained;

3 shows a greatly enlarged schematic representation of a micro servo ir according to the invention in vesicular form;

4- a greatly enlarged schematic representation a microreservoir according to the invention in spruce vesicular form;

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ORIGINAL INSPECTED

EIg. 5 is a greatly enlarged schematic illustration
a drug delivery vehicle of the known type;

6 is a diagram showing the in vitro stability of the microreservoirs in contrast to that of the
Liposomes explained, the in vitro stability being measured as the optical density of the microreservoirs and the liposomes as a function of the
Time;

7 is a diagram showing the in vivo stability of the
Microreservoirs, applied as the amount of
Chalesteryl oleate (cholesterol ester) retained in rat blood plasma as a function of time;

Fig. 8 is an elution profile of microreservoirs containing

Contain adriamycin, applied as phosphatidylcholine, Cholesteryl oleate and adriamycin concentrations in a range of chromatographic Fractions;

Fig. 9 is a schematic diagram showing the distribution of a drug such as adriamycin for short
after the injection of free drug, which indicates how little drug is in the bloodstream for delivery or delivery
the target center remains;

Fig. Ί0 is a diagram which explains the extent to which the circulating microreservoirs are able to are to change the plasma kinetics of adriamycin in an advantageous manner, the illustration in the form of diagrams in which the

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amount of adriaiaycin remaining in the blood stream as Function of time is plotted .. after, the injection of the drug into the microreservoirm and as free adriamycin;

Figure 11 is a series of graphs of the number of survivors a group of mice as a function of time after injection of Kikroreservoirs in four doses of adriamycin in the microreservoirs and adriamycin are administered in free form, which increases the ability of the microreservoirs explain, as a delivery or delivery vehicle act to reduce the toxicity of adriamycin;

Figure 12 is a series of graphs of the number of survivors of a group of mice exposed to tumor cells as a function of time after injection of two doses of adriamycin into the Micro reservoirs and in free form, polygamy Explain the beneficial modification of the chemotherapy of adriamycin, which is produced by the microreservoirs has been supplied and released therefrom;

Figure 13 is a series of graphs of discharge rates of adriamycin as a function of time from microreservoirs of a composition which G-lycerin trioleab as with the phospholipid no miscible ingredient and various amounts of cholesterol as a release rate control agent contain;

14 shows a series of diagrams of output and output

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Adriatic flow rates as a function of time from microreservoirs with a composition containing cholesteryl oleate than with the phospholipid immiscible ingredient and. various amounts of cholesterol as a release rate control agent contain;

Fig. 15 ^e iii elution profile of "microreservoirs containing AD 32, applied as a phospholipid, and Cholesteryloleat- AD 32 concentrations in a number of chromatographic fractions;

Fig. 16 is a graph of the discharge rate of AD 32 from Tesicular microreservoirs, which the Influence of using a smaller amount of phosphatidic acid in the phospholipid component the microreservoir composition shows;

Fig. 17 is a diagram explaining the extent up to which the circulating microreservoirs in are able to measure the plasma kinetics of AD 32 in to change advantageously;

Figure 18 is a series of graphs of the number of survivors a group of mice with tumor cells as a function of time after injection of AD 32 in the micro-reservoirs in four doses, which the chemotherapeutic effect of AD 32, which is supplied or fed through the microreservoirs is released, explain;

Figure 19 is an elution profile of microreservoirs containing Contain imidocarb, applied as a phospholipid, Cholesteryl oleate and imidocarb concentrations in a range of chromatography

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fractional;

Figures 20 and 21 are diagrams illustrating the extent to which the circulating micro-reservoirs are able to increase the plasma kinetics of imidocarb in two doses in a beneficial manner change, the representations being in the form of diagrams showing those in the bloodstream remaining amount of imidocarb and the ratios of imidocarb to the microreservoirs as a puncture of the time after the drug was injected into the microreservoirs and as a free one Imidocarb are applied;

S ¹ Ig. 22 is an elution profile of microreservoirs containing estradiol undecanoate plotted as phosphatidylcholine and estradiol undecanoate concentrations in a series of chromatography fractions;

Figure 23 is a bar graph showing the concentrations of estradiol undecanoate in blood plasma at three different time points, which is orally in In the form of an ethanol solution and administered in microreservoirs;

24 shows an elution profile of vesicular and non-vesicular microreservoirs, which contain estradiol undecanoate, and

Fig. 25 is a graph showing the rate of discharge of Estradiol undecanoate from vesicular and non-vesicular microreservoirs.

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ORIGINAL INSPECTED

The xenobiotic delivery vehicles of the invention are formed from a mixture of two or more lipids, which are essentially immiscible with one another. The supply and delivery systems are formed in particular from compositions containing a phospholipid ingredient and contain an ingredient immiscible with the phospholipid which is a gholesterol ester or a triglyceride or a mixture of cholesterol esters or triglyceridides or combinations both can act. The phospholipid component and the microreservoir composition can contain more than one phospholipid can also have a binding-modifying component and / or a component that controls the rate of breakage Part included. Examples of phospholipids that are useful in the practice of the invention are suitable are phosphatidylcholine, phosphatidylserine, Phosphatidylinosit., '. Phosphatidylethanolamine, Phosphatid- acid and sphingomyelin. It can also be mixtures of two or more of these phospholipids are used under Of these compounds, phosphatidylcholine either alone or in admixture with other phospholipids is preferred .

A preferred concept of the invention resides in a vehicle (a vehicle) that (s) for the introduction of a xenobiotic in a mammalian host is capable of increasing the pharmacodynamic properties or conditions (e.g. plasma kinetics, chemotherapeutic efficacy, toxicity, oral absorption, distribution within the tissue, the metabolism and the like) of the xenobiotic in an advantageous way. The supply bzx «i. Delivery vehicle lies in

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Form of microreservoirs made from a phospholipid component and a component which is not miscible with the phospholipid can be formed. Xenobiotic binding agents can also be used and release agents are added. Another preferred idea of the invention lies in processes for preparing the microreservoirs containing the xenobiotic and for administering one Xenobiotic to its pharmacodynamic properties to change in an advantageous manner.

As used herein, "phosphatidylcholine" refers to compounds that are esters of fatty acids trades in glycerophosphoric acid and choline. This phosphatidylcholine can be represented by the following formula:

FA-CH ₂ O

FA ¹ ₂

where FA and FA ^{1 represent} fatty acid residues. The fatty acids used to make these esters can be saturated or unsaturated and they can contain from about 12 to about 20 carbon atoms. These fatty acids include, without the invention being limited thereto, palmitic, stearic, oleic acid and the like Litman in "Biochemistry", 1L2, 254-5 (1973), isolated or derived from other natural sources such as soybeans, or

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it can be synthesized using an appropriate method such as "for example Robles and van der Berg in" Biochim. Biophys. Acta ", 187 , 520 (1969).

In the following detailed description of the invention, the phospholipid constituent of the Zuführungsbzw. For the sake of simplicity, delivery vehicles using Phosphatidylcholine explained, under this expression all esters are to be understood which are under those given above general formula fall. In some of the micro reservoir formulations lesser amounts of phosphatidic acid are incorporated into the phospholipid components to facilitate the bond of the xenobiotic to improve the microreservoirs. Of course, other phospholipids can also be used including those mentioned, which have the following physical and chemical properties, can be used in place of phosphatidylcholine.

As a component of the delivery vehicle used cholesterol ester or triglyceride component must be essentially immiscible with the phospholipid component and it must be in an aqueous solution Environment, represented by a physiologically compatible fluid, such as B- a physiologically balanced one Saline solution containing NaCl or KCl must be essentially insoluble. The cholesterol ester or triglyceride component must also be apolar or non-polar to such an extent that it does not form a monolayer, and it must be present in such a concentration that it is essentially immiscible in the phospholipid bi-layer is. .

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Cholesterol C ₂₇ H ^ ₁ -OH is a monounsaturated secondary alcohol which easily forms esters with both saturated and unsaturated fatty acids such as oleic acid, stearic acid, palmitic acid and the like Preferred methods for forming the cholesterol esters include condensation of a fatty acid chloride with cholesterol and isolation from natural sources.

When choosing the fatty acids for the preparation of the Cho Ie sterine st he s, it is preferred that one such with a lower degree of unsaturation (for example with no more than about two double bonds per fatty acid). In general, those form from fatty acids with a higher degree of unsaturation formed esters xenobiotic supply or release complexes with greater stability than those made from highly unsaturated Fatty acids are formed.

Those suitable for practicing the invention Triglycerides are fatty acid esters of glycerin with the general formula

H ₂ COCE ₁

0
HC-OCR ₂

0
H ₂ COCR ₅

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where E ^, R ~ ^vja - ^ ^ x of the fatty acids forming the ester can contain 10 to 18 carbon atoms. Examples of such fatty acids are palmitic acid, stearic acid, myristic acid, oleic acid and linoleic acid. These triglycerides are expediently produced by condensation of the fatty acid chloride with glycerol. They can also be isolated from natural sources.

In the following general description, for the sake of simplicity, it is assumed that a gholesterol ester is used. Of course, it is to be understood that in making the microreservoir delivery vehicles also a triglyceride, a mixture of gholesterol esters, a mixture of triglycerides or a mixture of one or more cholesterol esters and one or more Triglycerides can be used. There are below Also examples are given in which a cholesterol ester or a triglyceride is considered to be with a phospholipid immiscible ingredient is used.

The xenobiotic delivery vehicles of the invention are in what is referred to herein as "microreservoirs". These microreservoirs can have one of two shapes, namely a vesicular shape that has a thin-walled cavity that is used to manufacture of the microreservoir contains the medium used, or a non-vesicular form which contains the gholesterol ester and / or contains the triglyceride within the phospholipid monolayer. As described in more detail below, determines the ratio of phosphatidylcholine to cholesterol ester, which of these forms in any one Synthesis process predominates. The choice between these

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Form depends in principle on the type of xenobiotic, that is to be supplied and released, the pharmacodynamic Properties of the desired xenobiotic and the route of administration.

Two different synthetic routes for the production of the microreservoirs are shown schematically together with the incorporation of the xenobiotic in the same in FIGS. 1 and 2, the synthesis route shown in FIG. 1, in which an ultrasonic treatment for the production of the microreservoirs is used, is a preferred method. In this process, the phosphatidylcholine are used and the gholesterol ester in an inert organic liquid that is a solvent for both Components, such as B. chloroform, mixed together and if desired, the xenobiotic is like added to this solution indicated by the dashed line in FIG. 1. The solvent is then through Evaporation removed in vacuo leaving a dry residue mixture which can then be removed by adding a physiologically compatible liquid, for example an aqueous solution of NaOl or KCl of a suitable one Concentration, and a buffer is hydrated. If the xenobiotic is not added to the initial mixture, it is added to the resulting liquid suspension before the production of the microreservoirs.

When preparing the liquid suspension, it is preferred to use amounts of residue mixture which are equivalent to about 1 to about 5 ^{% by weight of} the physiologically acceptable liquid. Although the amount of the phospholipid / cholesterol ester (or triglyceride) -together-

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The amount of xenobiotic added can vary, the xenobiotic is incorporated into the microreservoiie iii, an amount that is preferably up to. about 2 ^, based on the weight of the residue. ^:

As explained in FIG. 1, the micro-reservoirs then formed by ultrasonic treatment of the saline solution (saline solution) in a non-oxidizing, for example in a nitrogen atmosphere. It has been found that a more complete formation of the micro-reservoirs is achieved will when the ultrasound treatment at a temperature is carried out, the 4em melting point of the cholesterol ester or Trigljrceridsj the bfcw. the fiCLs with the phospholipid immiscible component serves, corresponds to or is slightly above. The ultrasound treatment is performed at a suitable energy level and for a suitable period of time to achieve the desired Achieve microreservoir size. So, for example, has an energy supply of 120 W for a period of 20 minutes proved to be satisfactory. As an alternative to ultrasound treatment, the. . Salt suspension are pressed through a narrow nozzle to produce the microreservoirs.

As can be seen from FIG. 2, there is another type the production of the microreservoirs in an aqueous medium, for example in a \ saline solution, therein, a Prepare a solution of the phospholipid cholesterol ester and the xenobiotic using one with water miscible organic solvent, this solution then being converted into an aqueous physiological. \ 'Saline solution is introduced. Alternatively, the xenobiotic can be the

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ORIGINAL INSPECTED

- ■ "-st

. ■ 'Saline solution can be added prior to the injection step. An example of this method is the injection of an ethanol solution of the lipid mixture into a stirred aqueous one Solution or the slow injection of an ethereal solution of the lipid mixture into an aqueous compartment.

The cloudy liquid obtained in the formation of the micro-reservoirs in the aqueous suspending medium then becomes centrifuged to produce a clear phase and an upper cloudy phase, the latter being larger non-vesicular microreservoirs, for example those with a diameter of about 300 to about ΊΟΟΟ A contains. the clear phase is chromatographed to produce a fraction of small non-vesicular reservoirs (e.g. with a diameter of about 250 Å) and vesicular microreservoirs with diameters from about 190 to about 300 I.

The distribution of the microreservoirs between the vesicular and non-vesicular shapes is determined by the molar ratio from phospholipid to component immiscible with the phospholipid. When using phosphatidylcholine and alcohol ester oleate as an exemplary microreservoir composition can be shown to use from about 67 to about 97 mole percent phosphatidylcholine predominantly leads to the formation of vesicular microreservoirs, while the use of more than about 50 mol% Cholesterol esters for the predominant formation of non-vesicular microreservoirs leads.

The microreservoirs according to the invention have unique structural properties through the use of a

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Mixture of the phospholipid component and a lipid component (cholesterol ester, triglyceride or mixtures thereof), which with the phospholipid essentially does not,
is miscible. This structure is yesicular and in its
Non-vesicular shape shown schematically in Figures 3 and 4-. The insolubility of both the phospholipids (represented by phosphatidylcholine in Figures 3 and 4) and the immiscible lipid component
(represented by a cholesterol ester) leads to ■
Formation of an organized micro-reservoir structure, in
which avoids or minimizes contact between the immiscible lipids and the aqueous environment
is reduced. This organization in turn gives the micro-reservoirs a much higher stability than they do
is reached by a vehicle that continues after
above-mentioned known method has been prepared by Gregoriadis. This can be shown by a comparison between FIGS. 3 and 4 on the one hand and FIG. 5 on the other hand, FIG. 5 being shown schematically
Manner represents the structure of the drug delivery system according to Gregoriadis. From Figure 5 it can be seen that when miscible lipids are used to make the liposome, the result is a lamellar structure which leads to unstable organization, apparently because of the oxidation of the phospholipids which
destabilized the lamellar organization, or because of the
Crystallization of the phospholipids when the temperature drops below their transition temperature.

The stable structure of the microreservoirs according to the invention, as shown in FIGS. 3 and 4, can be used at least for
Part traced back to a thermodynamic driving force

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ORIGINAL INSPECTED

which prevents the cholesterol ester or exposing the triglyceride to the aqueous environment. The consequence of this is that the structure integrity of the invention Microreservoirs are greatly improved. This means that the ability of the microreservoirs to be pharmacodynamic Control and modify properties of the xenobiotics they contain, as well is improved compared to other supply or delivery systems. It also prevents apolarity the microreservoirs, which are a result of using cholesterol esters or triglycerides instead of Cholesterol or the like is essentially the aggregation of the microreservoirs. This is of course high desirable, since such aggregation would otherwise lead to a release of the gholesterol esters or triglycerides into one aqueous environment would result in a large decrease in the free energy of the system. the individual microreservoirs, which in many respects correspond to naturally occurring circulating serum lipoproteins are particularly well suited for circulation in the blood plasma of the mammalian host into which they are introduced and in which they remain in order to achieve maximum effectiveness.

The greatly improved in vitro stability of the microreservoirs according to the invention compared to that of the liposomes, as they have already been proposed, is j. _In the following example 1 and in FIG. 6 explained.

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example Λ

Various mixtures of 60 μmol egg yolk or egg yolk were used. Egg yolk phosphatidylcholine, various amounts of cholesteryl oleate and 4 mg of adriamycin from a chloroform solution of these Mur Reservoir components under vacuum into the Transferred dry. 5 ml of 0.1 M ECl and 10 mM trihydroxymethylamine (buffer from pH 8.0) was added and the resulting suspension was at 51 ° C under a nitrogen atmosphere for 15 minutes Treated with ultrasound at ¥ 120. Both suspensions were then spun at 100,000 g for 1 hour to un ~ to remove dispersed lipid (idpoid).

Thin layer chromatograms of the centrifugates showed no sign for a breakdown of phosphatidylcholine or adriamycin. Aliquots of 4 ml of each sample were made then placed in stoppered cuvettes and the samples incubated at 37 ° C. To different Points in time, the optical densities of the two solutions were determined at 400 nm and 600 nm. The received Dates are in the pig. 6 plotted as the ratio between these optical densities A 400 / A 600 versus time. Since both structures had practically the same molecular size at the beginning, the ratio A 400 / A 600 is an indication for an increase in particle size as a result of the breakdown of the structure of the liposome.

The Pig. Figure 6 illustrates the dramatic difference in stability between the microreservoirs of the invention and liposomes of similar size. While the extinction (the absorption capacity) of the 33 % cholesteryl

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oleate and small amounts of the cholesterol ester-containing microreservoirs were relatively stable, that of the liposome increased rapidly from the beginning of the evaluation period and after 10 days the liposomes had already degraded to such an extent that they could no longer be used in practice. As postulated above, it is assumed that the pronounced degradation of the liposomes is aurüek2ufüliren on the interaction with the aqueous environment and the aggregation to larger particles, two factors which are missing in the microreservoirs because of their structural stability and apolarity. In addition, the in-vitro stability of the microreservoirs in accordance with the requirements also showed a greatly increased in-vivo stability with respect to the liposomes. This see the following example 2 and Fig. 7-

Example 2

100 / aHol egg yolk phosphatidyl cliölin, 10 / µmol cholesteryl oleate, which had been marked with G, and 10 / µmol egg yolk phosphatidic acid were dissolved in chloroform and the solution was evaporated to dryness in vacuo. To the dry one mixed lipid residue was 5 eels of 0.154 M NaGl and 10 HiH trihydroxymethylamine (pH 8.0) were added. The one with it obtained suspension was at 51 ° C under a stick treated with ultrasound for 15 minutes. The mixture which had been treated with ultrasound was then chromatographed on a 2.55 ca.3 £ 4-0 es Sepharose-4E column. the individual fractions, ie a coincidence in the elution profile which showed hiospholipice and cliolesteryl oleate collected and then concentrated by ultrafiltration. 1.1 ml of these concentrated microreservoirs were added to the

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ORIGINAL INSPECTED

"" ** "" ¥ ί 291S028

Injected tail vein of a rat. It became plasma samples taken at different times and examined for the radioactivity associated with those in the plasma sample contained microreservoirs was related. The data obtained in this way are shown as a function in FIG after the injection.

Although the kinetics are complex, it can be seen from Figure 7 that at equilibrium, the clarity (margin) of the microreservoirs has a half-life of 5-5 hours. In contrast, the sonicated liposomes containing a similar negative charge have a plasma half-life of only 8 minutes (RL Juliano and D. Stamp, "Biochem. Biophys. Res. Comm.", 63 , 651 (1975)). It seems logical to conclude from this that the almost 40-fold increase in service life achieved by the microreservoirs is due to the increased structural stability.

The association of a xenobiotic which has been incorporated into the microreservoirs is explained in the following example 3 "οηύ in FIG. 8.

Example 3

2180 / Ufr.ol egg yolk phosphatidylcholine, 242 / U-Iöl cholesterol oleate, marked with a C (specific activi-

4 \

did 1.6 χ 10 disintegrations per minute (dpm) // µmol), and 36 mg Adriamycin were mixed together in chloroform and the solution was evaporated to dryness in vacuo. The mixed one dry residue was then added by adding

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110 ml 0.1 M KCl and 10 mM trihydroxymethylamine (pH 8.0) hydrated. This suspension was then taken at 51 ° C a nitrogen atmosphere for 15 minutes with a Branson W ~ 185 sonifer treated with ultrasound.

The liquid treated with ultrasound was then chromatographed on a 2.5 ~ 40 cm Sepharose-4-B column. The individual fractions obtained by chromatography were then separated by the method of Gomori ("J. Lab. Clin. Med.", £ 2, 955 (1949)) for their phosphate-1-dylcholine content and their cholesteryl oleate content was determined by radioactivity measurements and their adriamycin content determined by fluorescence measurements. The results of this analysis are shown in FIG. 8 in the form of a Diagram showing the analytical results for the cholesberyl oleate, for the phosphatidylcholine and the adriamycin are applied one on top of the other.

The diameter of the various vesicular microreservoirs (Bladder micro-reservoirs) were found to be about 200 to about 300 Å as determined by their elution profile a gel column previously made using sonicated liposomes of egg yolk phosphatidylcholine had been calibrated.

From Fig. 8 it can be seen that the elution profiles of phosphatidylcholine (monitored by phosphate analysis), des Cholesteryl oleate (monitored by radioactivity) and adriamycin (monitored by fluorescence) coincide, indicating that the xenobiotic (adriamycin) was associated with the vesicular microreservoirs. Because Adriamycin a relatively hydrophobic drug or

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29150.2 *

is a relatively hydrophobic drug with some properties, it is reasonable to postulate that that the place of localization of the drug Iszw. ster Drug lies within the lipid structure of the microreseriroire.

Among the pharmacodynamic properties of a Xeaolbioti-; Cums that can be changed to make the xenobiotic more effective include the plasma kinetics of each is -rad of toxicity and the therapeutic efficacy · Mria— mycin is regarded as one of the more effective eaeaotliei? It is actually relatively hydrophobic and it is very difficult to maintain a current in the flow, which is shown by the fact that by 2 minutes after the administration, about 95% of the blood flow had left the blood flow and was in various games had gone, with only about 5 % target tissue, δ. "H." a tumor. This is illustrated by Figure 9 which shows the release of adriamycin administered intravenously minutes after injection. Finally, adriamycin is known for tHe it is tösiseh, especially so because it feat the slope ₉ SiCl to concentrate in the heart tissues, it xrodurch _required; is not only to carefully surprise its administration, but also to administer it in very low dose valuesa.

In the examples described below A "to β TM in. FIGS. ΊΟ to 12, the ability of the e microreservoirs, which circulate in the blood stream, ÖIE plasma kinetics, the toxicity and the chemotherapeutic efficacy of adriamycin will change will be explained.

909850/0569
ORIGINAL INSPECTED

29 T 5Q28

Example * l

200yuMol egg yolk phosphatidylcholine, 20, uMd1 cholesteryl oleate and 4 mg of adriamycin, dissolved in chloroform and then transferred to the dryness under vacuum · To the thus obtained dry residue was added 5 nil 0.15 ¹ I- M Nagl and 10 mM Trihydroxymethylemin (pH This aqueous suspension was then ^{treated with ultrasound at 51 ° C. in} a nitrogen atmosphere for 15 "minutes 20 cm Sephadex G-50 gel column separated from the microreservoirs, and the void volume containing the adriamycin incorporated into the microreservoirs was reduced to 3 mg adriamycin / ml.

A control sample of free adriamycin in the same Concentration and in the same buffer was also made.

The microreservoirs containing adriamycin were intravenous 3 rats were injected and 6 more sats were injected intravenously with the free adriamycin. The dosage of the adriamycin contained within the microreservoir was 4 mg / kg and the dosage for the free adriamycin was 4 mg / kg in 3 rats and 10 mg / kg in the rest 3 rats. Su different points in time were made from each 0.5 ml blood taken from rat. The plasma was from the blood samples separated and examined for adriamyhin enamel fluorescence. The data obtained are shown in Fig. 10 plotted as a function of time after injection.

909880/0589

From the Pig. 10 shows that the use of circulating Microreservoirs as a delivery vehicle for the Adriamycin beneficially changes and improves the plasma kinetics for this drug. A comparison of curves A and B, which represent the same dose level, shows that after about 15 minutes the concentration of adriamycin in the circulating microreservoirs in the plasma was about 4 / µg / ml, while the concentration of free adriamycin in the plasma was about 0.4 / Ug / ml. That's why at that point the focus was on of adriamycin in the blood stream carried by the circulating microreservoirs about ten times larger than if this medicine or drug had been introduced directly into the bloodstream. Also goes out FIG. 10 shows that the free adriamycin was practically completely absent from the bloodstream after about 3 3/4 hours, while at this point it is still up to a concentration of about 0.4- / ug / ml in the circulating microreservoirs was included. The concentration of the drug in that carried in the circulating microreservoirs Blood flow after 3 3/4 hours corresponded to that after only 15 minutes if the drug is introduced in free form became.

A further comparison between curve A and curve G (free adriamycin at a dose of 10 mg / kg) shows that after 1 hour the concentration of the drug in the plasma was about 1.3 / Ug / ml when it was circulating from dsn Microreservoirs were worn, and that they -about 0.5 / Ug / ml when it was in free form; after 3 hours these numerical values were about 0.5 / µg / ml and about 0.3 / Ug / ml and after 6 hours sxb were about Ό, 21 / ug / ml and

9Q9850 / ÖS69
ORIGINAt INSPECTED

0.14yUg / ml The use of the circulating Nikroreservoirs according to the invention can therefore produce concentrations of this drug in the bloodstream which are significantly higher than those which are achieved when the same drug is administered in free form in dosages that exceed 2, Are 5 times larger than "when it is administered in the micro-reservoirs.

The change in the plasma kinetics of a drug, such as B. Adriamycin, as shown in Fig. 10, means that it has a lower toxic effect , since it is kept in the bloodstream for a longer period of time / and that it is thereby prevented that it is in the organs such as the liver, Kidney and heart, enriches. It also means that any given dosage will be more effective when administered in the form of the circulating microreservoirs than when the drug is freely circulating, since the more drug remains in the bloodstream, the more exposed the target tissue to the adriamycin dose. Because this drug's effectiveness in killing tumor cells relies on the tumor cells' ability to take up it, maximum contact between the drug and the cells is essential for maximum effectiveness.

Finally, changing the plasma kinetics of a xenobiobic, as shown in FIG. 10, offers the possibility lowering of doses; assuming, for example, that the 10 mg / kg dose of curve C for chemotherapeutic Purposes is to be regarded as sufficient, it follows from this that the dose value is reduced below 4 mg / kg can wens the drug out of the circulating

909850 / ΟΊ569

Microreservoirs released and released · Such a decrease in dose would also markedly reduce toxicity. That a significant reduction in adriamycin toxicity is achieved by using the circulating hikroreservoir is also evident from the example described below and FIG. 11.

Example 5

., 100 / uIiol Gholesteryl- oleate yolk Phosphatidylcholifl and 20 mg adriamycin was dissolved in ühlorofona and the solution was concentrated in vacuo to dryness. To the resulting dry Eückstandsmischung were added (pH 8.0) 20 ml 0.1 M KCl and 10 mti Trihydroxymethylamin. The aqueous suspension was treated with ultrasound for 15 minutes at 5 ° C. under a nitrogen atmosphere. The ultrasound-treated liquid was chromatographed on a 2.5 cm × 40 cm Sepbarose 4B gel column. Only those fractions were collected in which the elution profile of phosphatidylcholine »alcohol esteryl oleate and adriamycin coincided. These pooled fractions were then concentrated by ultrafiltration using a 2M-50 membrane to a final concentration of 3 mg adriamycin per ml.

A similar sample of microreservoirs devoid of adriamycin. was also made and concentrated to the same micro-reserve concentration. In addition, a solution of free adriamycin in a concentration of 3 mg / ml in 0.1 M KCl and 10 mH (urihydroxy- CpH 8, O) was prepared as a control, which was used as free adriamycin

909850/0569

ORIGINAL INSPECTED

was used·

The microreservoirs containing the adriamycin, the microreservoirs Without the drug and the free drug were taking in different drug doses - using administered to mice in a single intraperitoneal injection. 10 BDFxj mice were used for each experiment and the data thereby obtained are shown in FIG. The diagrams obtained show the survival rate of the home as a function of time. From this it can be seen that the microreservoirs showed no toxicity even in the absence of adriamycin. At all dose levels, eh were made using the circulating microreservoirs to transport the adriamycin the toxic effects of the drug compared to those that at which free adriamycin occurred, decreased. With decreasing dose levels, the ability of the delivery vehicle increased to reduce toxicity more pronounced compared to the free drug. It is believed, that. this reduction in toxicity, at least for the iDeil can be explained by the ability of the circulating microreservoirs to carry adriamycin in and out of the bloodstream of the tissues and organs, especially the heart. The effects of adriamycin chemotherapy in Use of the delivery vehicles according to the invention are described in the following example 6 and in FIG. 12 explained.

Example 6

The procedures of Example 5 were repeated for preparation of vesicular microreservoirs with and without adriamycin

909850/0569

for the manufacture of adriamycin for use as a free drug. A number of mice were ^{injected intraperitoneally with 1 × 10 6} P 388 tumor cells. Twenty-four hours later, single daily intraperitoneal doses of the vesicular microreservoirs with and without adriamycin and free adriamycin were injected after the tumor implants for a period of 5 days. A group of mice used as a control did not receive any drug. Five mice were used for each experiment and two different dose levels, 4 mg / kg and 2 mg / kg, were used. The data obtained in these tests are shown graphically in Figure 12 as the number of animals surviving as a function of time.

The mice injected with the microreservoirs without adriamycin had a survival rate (not shown) which was similar to that of the control mice, curve G in the graphs. At the dose of 4 mg / kg the adriamycin released by the circulating microreservoirs had a chemotherapeutic effect while the free drug at the same dose exhibited toxicity measurably greater than that of the Tumor cells. At the lower dose of 2 mg / kg this showed Adriamycin released by the circulating microreservoirs greater chemotherapeutic responsiveness stuf as the free drug,

When formulating the microreservoirs according to the invention may -be- one or more xenobiotics- binding modifiers -to incorporate. who-able the relative amount of xenobiotics taken up by the microreservoirs during their formation either increases

9O98SO / G569

increase or decrease. For example, it has been found that a lower mole percentage, i.e. H. about 5 mol% or more, phosphatide, acid, a charged lipid that corresponds to the Phospholipid component of the microreservoirs is added, the affinity of the xenobiotic for the microreservoirs can increase. It is therefore within the scope of the present invention to have one or more different ones Use phospholipids to supplement the phospholipid component of the microreservoir composition. It lies in addition, even within the general knowledge of the art, a single phospholipid or an optimal combination of phospholipids as a phospholipid component of the microreservoirs to be selected in such a way that a given intake © of the xenobiotic is achieved.

It is also possible to reduce the binding of a xenobiotic to the reservoir by incorporating other lipids that are in soluble in the phospholipid component or in the cholesteryl ester or triglyceride component slightly soluble are to be modified. Those in a microreservoir / adriamycin system Modifications achieved are described in the following Example 7 - and in the table I explainsj the Effect such modifications on xenobiotic delivery or the pricing rates resulting therefrom Microreservoirs are discussed in the following below Example 8 and illustrated in Figs. 13 and 14-.

Example 7

It became a hot topic of formulations of microreservoirs ^ the 1G5yUMX> l Egg Yolk Phosphatidylcholine, the

2916028

marked with G "(specific activity 4160 dpm / uHol phosphatidylcholine), 11.6 / uHol cholesteryl oleate or 11.6 / µHol trioleiii (slyceryl trioleate) with or without a second phospholipid., represented by 11.6 , µmol phosphatid · - acid, and with or without cholesterol as Modifying agent, in an amount from 0 to 75 / µmoles was present, contained. In each formulation, 1.72 mg of adriamycin was used as the xenobiotic contained therein was carried and released from it.

The formulations were prepared in the form of chloroform solutions, which were transferred to the dry state in 10 ml vials with the lid screwed on by pumping them out. Each formulation sample was then mixed with 3 ml 0.154 H NaCl and 10 mM trihydroxymethylamine (pH 7.2 ) and then stirred for several minutes at room temperature. Each sample was then sonicated at the appropriate temperature for 20 minutes in a stream of nitrogen. Sonication of these samples containing cholesteryl oleate was carried out at 5 ° C and that of those containing triolein was T ^: Carried out at 3 ° C. After the ultrasound treatment, each sample was spun in a bench-top centrifuge for 10 minutes in order to remove any titanium fragments therefrom.

J: Each sample was made using 0.154 "M * NaCl and 10 mM trihydroxymethylamine as elution buffer through a Run 2.5 cm by 15 cm Sephadex G-50 columns. That Void volume geder the G-50 columns, which the microreservoirs contained, was collected and applied to the yolk

14th
Phosphatidylcholine -C-Eadioactivity and the fluorescence of adriamycin. examined. The results of this

"9098-50 / 0569. ORIGINAL INSPECTED

- • Μ- -

Measurements expressed in yug adriamycin / µmol phosphatidylcholine and / Ug adriamycin / / µmol total phospholipid and the percentage of adriamycin uptake by the microreservoirs are summarized in Table I below. In these results, the percentage of encapsulated adriamycin was normalized with reference to the sample containing the highest level of encapsulated adriamycin per / µmole total phospholipid.

Table I.

Influence of the composition of the phospholipid component and the addition of phospholipid-miscible lipids to the adriamycin uptake through the microreservoirs

Microreservoir—
composition
Mole% GTO CO PA Chol / Ug Adriamycin / / µmol
⁷ pl % turned
encapsulates sample
No. PO 9 - 9 / µmol
Pc 6.03 100 1 82 10 - - - 6.67 4.90 81 2 90 9 - - 9 4.90 4.26 '71 3 82 8th - 20th 4.26 4.14 69 4th 72 7th - 28 4.14 4.03 67 5 65 6th - 40 4.03 3.91 65 6th 54 - - 9 - 3.91 5.27 87 7th 82 - 10 - - 5.83 4.41 73 8th 90 - 9 - 9 4.41 4.21 70 9 82 - 8th 20th 4.21 3.43 57 10 72 - 7th 28 3.43 3.14 52 11 65 6th 40 3.14 2.36 40 12th 54 2.36

909850/0569
ORIGINAL INSPECTED

PG - phosphatidylcholine

GTO - glycerine trioleate

CO - cholesteryl oleate

PA - phosphatide - acid

Choi - cholesterol

PL - total phospholipids (PC and PA)

From the data in Table I above it can be seen that by incorporating a small amount of phosphatidic acid into the phospholipid component (Samples 1 and 7) the Adriamycin uptake would have been increased by the microreservoirs. In contrast to imidocarb (Examples 13-15), adriamycin is an uncharged molecule, a fact that indicates that the use of phosphatidic acid as part of the phospholipid component on xenobiotics with various Properties can be applied.

7) ie incorporation of cholesterol, one with phospholipid miscible lipid, in the microreservoir composition prevented the adriamycin from binding to the microreservoirs. The addition of cholesterol to the microreservoir compositions, which contained glycerol trioleate (samples 3 to 6) had a significantly lower effect on the binding of the drug than in the case where the microreservoir composition contained cholesteryl oleate (Samples 9 to3 12). Finally, the data in Table I show that in the case of adriamycin, the use of glycerol trioleate (Samples 1 to δ) instead of cholesteryl oleate (Samples 7 to 12) the binding of this Xenobiotilcums to the

Microreservoirs have been facilitated. It appears, therefore, that the choice of the phospholipid component and that with Phospholipid immiscible ingredient (with or without an additional xenobiotic binding modifier) it is possible to control the degree of xenobiotic uptake or binding to the microreservoirs and to be determined beforehand. Such control gives the xenobiotic delivery system of the invention flexibility in the Regarding the dosage levels, the xenobiotic release rate and the same.

Not only can the equilibrium binding of a xenobiotic be influenced by the microreservoir composition, but the rate of release or release of the xenobiotic can also be controlled and predetermined by the composition. This is explained by using a model system which allows the xenobiotic discharge rates to be determined from the microreservoirs in accordance with the non-equilibrium conditions, as explained in more detail in Example 8 below and shown in FIGS. 13 and 14.

Example 8

As indicated in Example 7, microreservoirs were made. In order to establish the necessary non-equilibrium conditions, the Adriamycin were used as a xenobiotic retaining microreservoirs with a large excess of Egg yolk phosphatidylcholine dispersions not treated with ultrasound incubated. These dispersions were made by adding aliquots of

8 5t) / 05-6 9

Microreservoirs of various compositions that contained about 1 / µmole egg albumin phosphatidylcholine, too 9 / µmol non-sonicated egg yolk phosphatidylcholine in 1.0 ml of 0.154 H NaGl and 10 mM trihydroxymethylamine (pH 7j2). There were comparable control dispersions produced an equivalent amount of free adriamycin instead of that contained in the microreservoirs Adriamycins contained.

For determining the rate of discharge (outflow rate) of adriamycin became one of the so produced for the time being Mixtures used. At the indicated time, the sample was subjected to 2 minutes at 15,000 g under such conditions centrifuged, among which the non-sonicated dispersion slightly differs from that obtained in the supernatant Liquid remaining microreservoirs was separated. Since the adriamycin to re-equilibrate between the Micro-reservoirs and the non-sonicated phospholipid dispersions can reduce the rate of equilibration as a kinetic parameter for determining the role of the various components making up the microreservoir composition Components for determining the discharge rate or outflow rate of the adriamycin contained in the microreservoirs be used.

An aliquot of the obtained supernatant liquid was examined for fluorescence and the result remaining fluorescence was compared to the fluorescence originally present in the mixture. The amount of Adriamycin remaining in the supernatant fluid therefore represents the amount of drug that was still contained in the microreservoirs. The one with this one

909850/0569

The data obtained from the series of measurements are plotted in FIGS. 13 and 14 in the form of a graph showing the decrease in the Fluorescence as a function of time for the microreservoirs containing glycerol trioleate (FIG. 13) and the cholesteryl oleate containing microreservoirs (Fig. 14) show. From the data plotted in FIGS. 13 and 14 it can be seen that that the free adriamycin was removed from the supernatant fluid very quickly, while the Microreservoirs bound adriamycin was retained to a much greater extent. Incorporation of 9 mol% Phosphatidic acid in the phospholipid component of the Microreservoirs led to a substantial delay in the discharge rate or outflow rate during the addition of Cholesterol this increased. Eventually, the use of a triglyceride instead of a cholesterol ester led to a slight decrease in the delivery rate.

These kinetic ratings were based on delivery rates or outflow rates relevant to the data in the table I, which concern the equilibrium binding of adriamycin to the microreservoirs; confirm this data the fact that the composition of the micro-reservoirs can be selected for determination and control the pharmacodynamic properties of the xenobiotic contained in the microreservoirs, these as serve according to the invention drug delivery vehicle. AD 32 is a highly hydrophobic analog of Adriamycin, which is used in chemotherapy. As this drug is completely in an aqueous buffer is insoluble, a combination of a detergent and an organic liquid must be used for preparation a solution of AD 32, intended for clinical

909850/0669 ORIGINAL INSPECTED

Use is suitable. An example of such a thing Solvent currently in use is a mixture of equal volumes of ethanol and a sulfated one Ethylene glycol detergent (Emulphol). Despite the hydrophobic nature of AD 32, it is possible to incorporate it into the invention Incorporate microreservoirs and thus open beneficial way of its pharmacodynamic properties with respect to both plasma kinetics and therapeutic efficacy, as in the following Examples 9 to 12 and in the Pig. 15 to 18 explained.

Example 9

1Q90 / µmol phosphatidylcholine from egg yolk, 121 / µmol gholesteryl oleate marked with ^ H (specific activity 190,000 dpm / µmol) and 17.5 mg AD 32 were dissolved in chloroform, then transferred to dryness and pumped off overnight. 5 »5 nil 0.154 M NaGl and 5 mM trihydroxymethylamine (pH 7, -4-1) were added to the dry mixture. The liquid was treated with ultrasound under a nitrogen atmosphere at 48 ° G and the resulting ultrasound-treated liquid was chromatographed on a 2.5 cm χ ¹ V cm Oepharose ^ B column. The individual fractions obtained in this way were examined for their phosphate / dyl content, for their content of cholesteryl oleate and of AD 32, as described above. The resolution profile of a line-up of the analysis values int is shown in FIG. 15 in the form of a diagram. From this diagram it can be seen that the AD 32 'has both non-vesicular hicroreservoirs

(Fractions 2 to 5) as well as with vesicular microreservoirs (Fractions 6 to 18) is associated.

The incorporation of a lesser amount of phosphatidic acid into the phospholipid component of the microreservoir composition has, as has been found, only a minor, if any, effect on the rate of discharge (outflow rate) of AD 32 when a triglyceride was used as an ingredient immiscible with a phospholipid. This is possible from Example 10 below and FIG. 16.

Example 10

A basic microreservoir formulation was used which contained 104.6 / µmoles of egg yolk phosphatidylcholine and 11.6 / µmoles of glycerol trioleate which had been labeled with H (specific activity 1.5 x 0μ dpm // µmoles). 1.73 mg of AD 32 were added and, in one formulation, 11.62 / uHol egg yolk phosphatidic acid were also added. These formulations were transferred to dryness from a chloroform solution and pumped out under vacuum overnight. To each sample, 3 ml of 0.154 M ITaGl and 10 niM trihydroxymethylamine were added and the hydrated fluids were sonicated for 20 minutes at 3 G under a nitrogen atmosphere. Each sample was passed through a 2.5 cm x 20 cm O8phadex C5-50 column and the amount of each vesicular fluid sample obtained, which was equivalent to 1 / µl of phospholipid, was mixed with 9 / µM of phosphatidylcholine -Liposoniön incubated in 0.46 ml of a buffered saline solution to produce a dispersion.

909850/0569

The Centrifuged dispersions for 2 minutes at 1500 g and the fluorescence remaining in the supernatant was determined. The data from these measurements of the Discharge rate (outflow rate) are shown in Fig. 16 in the form of a Indicated in the diagram. From this it can be seen that the delivery rate of AD 32 was relatively high and that it by incorporating phosphatidic acid into the microreservoir composition remained essentially unaffected. Although the rate of release of AD 32 from the micro-reservoirs is relatively high, the use of microreservoirs leads to a significantly more favorable change the plasma kinetics of this drug compared to the dosage forms previously used. This is done in. the following Example 11 and FIG. 17.

Example 11

Fractions 6 to 18 of Example 9 were passed through Ultrafiltration up to an AD 32 concentration of 3 »2 mg / ml concentrated. Sufficient quantities of AD 32 indicated in the microreservoirs intravenously 150 g rats to achieve a dose of 15 mg / kg; and a clinical formulation of AD 32 dissolved in a 1: 1 volume mixture of ethanol / emulphol was injected in a corresponding manner in the same dose 3 control rats adorns. Plasma samples were taken at various times and the AD 32 concentrations in these plasma samples were determined by fluorescence. The data thus obtained (average of three rats for each Point) are shown in FIG. 17 as AD 32 equivalents as

909850/0569

Function of the time from the start of the injection in the form of a Indicated in the diagram.

Fig. 17 shows the marked change in plasma kinetics of AD 32, achieved by using the microreservoirs according to the invention, since the contents of AD 32 in the plasma in which the drug was incorporated into the microreservoirs, at all times by a factor between 2 and 3 were higher than the levels of free AD 32, which was introduced as a clinical solution became. By letting the drug seek out tumor cells for a longer period of time, they become Change of its plasma kinetics its effectiveness as chemothei "apeutisches Agent improved. This is also shown by the following example 12 and FIG. 18.

Example 12

As in Example 9, vesicular hicroreservoirs containing AD 32 were made and concentrated to 3j2 mg AD 32 / ml. A number of mice were injected intraperitoneally with 1 × 10 P3S8 tumor cells (leukemia cells). 24 hours later, single daily intraperitoneal doses of the vesicular microreservoirs with AD 32 and Saline control solutions are injected for a period of 5 days after the tumor implants. For every attempt 5 mice and 4 different doses, namely 7 »3 mg / kg, 4 mg / kg, 2 mg / kg and 1 mg / kg were used. The ones with these Data obtained from tests are shown in Fig. 18 in the form of a graph as the number of survivors as a function of the number of survivors Time applied. In all cases, the mice pointed out that

909850/0569

the microreservoirs containing AD 32 were injected
had a higher survival rate than the comparison mice (control mice). In addition, self- '
the very low '. Dose of 1 mg AD 32 / kg delivered
through the circulating microreservoirs, to a chemotherapeutic effect and to a significant increase in the survival rate of the mice administered with the P388 leukemia cells.

As in the case of adriamycin, the same plasma concentrations of AD 32 can be achieved at lower starting doses of
AD 32 contained in the microreservoirs can be achieved than with clinical formulations. Furthermore
the microreservoir composition is more biologically compatible with blood than the mixed solvent
Ethanol and detergent required in AD 32 clinical formulations today.

Adriamycin and AD 32, a well-known garminomycin, are cancer chemotherapeutic agents that belong to the general class of authracyclin drugs. The preceding examples illustrate the effectiveness of the delivery yehiculum according to the invention in terms of beneficially modifying the pharmacodynamic properties of this class of chemotherapeutic agents. The microreservoirs according to the invention can also be used to dispense
other classes of chemotherapeutic agents, such as
B. nitrosoureas and metaboutes can be used.

Imidocarb is a medicine that has proven to be a very effective parasiticide in the treatment of anaplasma in animals by killing the parasites in the bloodstream

909850/0569

Has. When administered in its most effective doses, however, trace amounts of the imidocarb remain in the animal tissue, an undesirable situation when that occurs Animal is intended for human consumption. Therefore, it would be desirable to have a delivery vehicle available too which the imidocarb can hold in the circulating blood stream so that it can achieve its therapeutic effect can unfold without accumulating in the animal tissue.

Imidocarb and its hydrochloric acid salt are hydrophilic compounds, a characteristic which distinguishes them from hydrophobic or hydrophobic / hydrophilic drugs. As from the below described Examples 13 and 14 and Figures 19-21. As can be seen, the microreservoirs are the same Effective in terms of the beneficial change in the pharmacodynamic properties of a hydrophilic xenobiotic like imidocarb.

Example 13

900 / µmol egg yolk phosphatidylcholine, 100 / µmol phosphatide- acid, 100 / µmol cholesteryl oleate, which was marked with "Έ (specific activity 1.6 χ 10 dpm // µmol), and 9.16 mg

14th
Imidocarb marked with C were dissolved in chloroform and the solution was evaporated to dryness in vacuo. The resulting mixed dry residue was then hydrated by adding 45 ml of 0.154 M NaCl and 290 mM trihydroxyethylamine (pH 8.0) to the mixed lipid / drug residue. The liquid was

909850/0569

& 3 2315028

then treated with ultrasound for 15 minutes at ^ Λ ° Ο under a nitrogen atmosphere. The liquid treated with ultrasound was then chromatographed on a 2.5 cm × 40 cm Sepharose 4B column. The individual fractions obtained during the chromatography were then analyzed for their phosphatidylcholine and phosphatidic acid content by the Gomori method investigated and you

■ z content of cholesteryl oleate, which was marked with ^ H, and

14th
of imidocarb marked with C was determined by radioactivity. The results of these analyzes are shown in FIG. 19 as a function of the fraction number in the form of a diagram, with the analysis results being plotted on one another in FIG. 8. The diameter of the vesicular microreservoirs was found to be within the range between about 200 and about 300 pounds.

From this it can be seen that the elution profiles of the three Components of the microreservoirs in fractions 5 to 9 coincided, indicating that the imidocarb was associated with the micro-reservoirs. The beneficial change the plasma kinetics of imidocarb achieved by using the delivery vehicle according to the invention is shown in the following Example 14 and Figs. 20 and 21.

Example 14

100 / µmol phosphatidylcholine and 10 / µmol phosphatide: -. Acid from egg yolk, 10 / µmol cholesteryl oleate and 2 mg imidocarb,

i4
marked with C, were dissolved in chloroform and the

909850/0569

The solution was evaporated to dryness in vacuo. The one with it mixed residue obtained was dissolved in 5 ml of 0.154-H NaCl and 10 mM trihydroxrmethylamine (pH 8.0) suspended and the resulting suspension was at 51 G under a Ultrasonically treated nitrogen atmosphere for 15 minutes. The liquid is then poured out on a 2.5 cm 40 cm Sepharose 4B gel column and the

14th
Fractions containing the imidocarb marked with C, phospholipids and cholesteryl oleate were collected and concentrated by ultrafiltration to a final concentration of 0.6 mg imidocarb per ml.

In addition, a solution of free imidocarb at a concentration of 0.6 mg / ml in 0.154 M HaGl and 10 ml'l trihydroxymethylamine (pH 8.0) prepared the was used in the free imidocarb administration.

The imidocarb containing hikroreservoirs and the free one Imidocarb was injected into the swan vein of rats in two doses; H. at doses of 5 mg / kg and A, 4 rag / kg. Plasaic samples were taken at different times and the amount of laidocarb in these samples was taken determined by measuring their radioactivity. The results of these measurements plotted as imidocarb equivalents as a function of time are given in FIGS.

The beneficial change in plasma kinetics through use the circulating hikroreservoirs that contain the imidocarb transported in the bloodstream of a living mammalian host, is clear from FIGS. 20 and 21. For example, at a dose of 50 / kg that was

909850/0569

2315028

Concentration of the imidocarb in the microreservoir / imidocarb in the bloodstream was about 140 times that of the free imidocarb after 4 hours, and at a dose of 4.4 mg / kg it was about 200 times higher after the same period of time. In addition, as indicated in FIG. 21, the blood stream still contained a measurable amount of imidocarb after 24 hours in the form of microreservoir / iniidocarb in contrast to the free form ύοώ. Imidocarb, which was practically no longer present.

The ratio of imidocarb to microreservoirs as a function of time is also shown for each dose in FIG and 21 plotted; these graphs clearly show that the drug is being released at a rate suitable for any predetermined conditions can be controlled and specified in advance. From FIGS. 20 and 21 is also possible indicate that it is possible, if desired, to reduce the doses of the drug to levels well below those values that were previously considered to be effective in killing the parasites swimming in the blood, the anaplasma cause in animals.

One of the main goals in beneficially altering the pharmacodynamic properties of a xenobiotic is the ability to change the distribution of the xenobiotic in the tissue. This can be achieved for imidocarb by using the microreservoirs according to the invention, as indicated in the following Example 15 ua-d in Table II.

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2915Ü2S

Example 15

The animals used in Example 14 were 4 hours after the injection of free or imidocarb containing imidocarb. Microreservoirs killed. Xrarden from the animals various fabrics such as B. muscle, spleen, liver and kidney gex '/ ebe removed. These tissues come together in one Searle incinerator burned and the

14th
Any COp held was collected and radioactivity was counted. The number of counts per gram of tissue was determined and the results are given in Table II below.

Table II

Distribution of free imidocarb and microreservoir imidocarb in the tissue

- dpm / g tissue -

tissue Imidocarb Imidocc-rb 'Change m% muscle 3985 3511 -12 kidney 40194 59984 +49 spleen 9498 15 339 +61 liver 27059 29991 +11

The ability to hold a xenobiotic in the bloodstream and within certain organs is particularly significant with a drug such as iraidocarb. Reducing dec, imidocarb in the muscle tissue by 12 % caused by

909850/0569

Using the microreservoirs is x-correct for the use of this parasiticide in the treatment of animals intended for human consumption? are. In addition, the higher the concentration of the drug in the liver and in the spleen, in organs that contain a greater proportion of the anaplasma microorganisms included, another important parameter when taking advantage of the use of imidocarb.

On the pharmacodynamic properties of a xenobiotic, which can be changed in an advantageous manner by the Abgabe-Vehiculu.ni according to the invention, belongs oral absorption, the modification of which is achieved by giving the xenobiotic the ability to Gastrointestinal (GI) tract to pass and for that effective circulation to enter the bloodstream. There are currently several solutions for the administration of xenobiotics available which do not pass the G-I tract or which pass it to a very limited extent and get into the bloodstream. One such solution is to chemically modify the xenobiotic, for example by forming the undecanoate ester of estradiol, which is practically insoluble in water, and suspending it the same in oil, the transformation of the same into a microcrystalline Dispersion or the preparation of a solution with an organic solvent such as ethanol. The training of estradiol undecanoate (used as a fertility control agent is used) in the micro-reservoirs according to the invention makes the use of such liquid media superfluous as oils and organic solvents, and at the same time it becomes a much more satisfactory one Release of this xenobiotic into the bloodstream

90 9 8 50/0569

achieved. The incorporation of estradiol undecanoate into the microreservoirs according to the invention is explained in the following example in FIG. 22.

Example 16

500 µmoles of egg yolk phosphatidylcholine, 30 µmoles of cholesteryl oleate and 6 µmoles of estradiol undecanoate labeled with ^ H (specific activity 4 ₃ 6 µCi // µmoles) were dissolved in benzene and lyophilized. 5 ml of 0.154 M NaCl and 5 mM trihydroxymethylamine were then added to the dry mixture obtained in this way and the liquid obtained was treated with ultrasound at 48 ° G under a nitrogen atmosphere for 17 minutes Fractionated chromatography on a Sepharose 4-B column, giving the elution profile according to]? Ig The ability of the microreservoirs of the invention to increase the oral absorption of estradiolun decanoate is further illustrated in Example 17, below, in Figure 23 and in Table III.

Example 17

Estradiol undecanoate marked with * H was placed in microreservoirs encapsulated, which consisted of egg yolk phosphatidylcholine and cholesteryl oleate labeled with C.

909850/0569 ORIGINAL INSPECTED

Equivalent amounts of ^ H labeled estradiol undecanoate were dissolved in ethanol. Doses of 1.63 mg / kg were orally administered to two groups of 9 rats. After 1, 4 and 24 hours, three of the rats in each Group killed and the blood was collected. The plasma was removed from the robotic blood cells by centrifugation separately and 0.5 ml of each plasma sample became 9.5 & 1 CHCiyMeOH (volume ratio 2i1) added, the 1.6 .mol contained unlabeled estradiol and estrone as carriers. The resulting liquid was filtered and added to the Filtraten xmirde enough saline solution added to a 2-phase system. The lower phase becomes Concentrated to dryness and then redissolved in 2.0 ml of methanol. 0.2 ml of this solution were used to determine the Radioactivity counted. The remaining 1.8 ml were applied to a TLG plate in the form of dots and in Bensol / ethyl acetate (volume ratio 3: 2) developed. The plates were briefly exposed to iodine vapor to remove the carrier steroids visible, and then the iodine vapor was driven off by gentle heating. The stains that Estradiol and oestrone corresponded, were excised and counted to determine their radioactivity. the Are equivalents of - ^ H-estradiol undecanoate in the plasma plotted in FIG. From this it can be seen that in In the first hour the vesicular iliac reservoirs the occurrence of estradiol undecanoate equivalents marked with ^ H in the plasma increased compared to the ethanol control solution. After 4 hours the amount of estradiol undecanoate equivalents decreased in both formulations.

It is known that estradiol is the active form of the drug and estrone is inactive. It is also assumed

9Ώ9850 / 056S

that the estradiol esters are inactive until hydrolysis to estradiol. Therefore, the ratio of estradiol to estrone in the plasma should be an index of the ability of the microreservoirs to improve (increase) the active form of the drug. The results of TLC analysis of the labeled estradiol derivatives in the plasma are given in Table III below. From these data it can be seen that the ratio of estradiol to estrone is 60 % higher when using the microre servo ire than when using an ethanol solution.

Table III

Ratio of ^ !! - estradiol to H-estrone in the plasma 1 hour after the administration of oesbradiol undecanoate

Eorration oestradiol / oestrone *

in ethanol solution 1.14

in microreservoirs 1.84

* The ratio was based on the mean of 3 rats.

The microreservoir delivery system according to the invention also offers the possibility of oral administration of xenobiotics, which have not yet been administered in this way could. By transporting these drugs through the GI tract, it is possible to get them indirectly into the blood stream introduce, thereby increasing their metabolism in the GI tract

90 9 85 Π / 0569

It is avoided and it is more necessary to sting them
inject and the disorders _; and accept complications associated with this form of administration. , -: '

The microreservoirs according to the invention can be used in their vesicular form or in their non-vesicular form or in a combination of these forms. this
is illustrated by the following example 18 and FIGS.

Example 18

Λ1

25 / µmol egg yolk püiospnatidylcliolin, 75 / ^{UI 11} IoI caolesteryl oleate labeled with C, and 2 aMoI - ^ Η-labeled estradiol decanoate are dissolved in chloroform . in the
! Dry transferred. The lipid mixture was dried overnight in vacuo and then with 4.4 ml of 0.154 M MaCl and
5 mM trihydroxymethylamine (pH 7.2) hydrated. The liquid obtained in this way was treated with ultrasound for 20 minutes under a nitrogen atmosphere at 40.degree. C. and passed through a Sepharose 4B column. The elution profile obtained in this way is shown in FIG. 24. It can be seen from this that the estradiolunde anoate has a higher affinity for the vesicular form of the micro-reserves than for the non-vesicular form, based on the ratio of estradiol anoate to
Cholesteryl oleate in the two forms of microreservoirs. The material in the void volume of the uoloiine consisted of the non-vesicular form, while the material in the internal volume consisted of the vesicular form.

909850/0569

ORIGINAL INSPECTED

291S028

• "'Prosen the

the "-ösijradiol-undecHiioat Bad" twa 1-yuIiol Gesastlipid contained © ^,. WW3PÖ8B 'znasaamen with "9 / dlol eini? not with ultra . scMö.11'.behenöelten Bigs ^ fe-I ^ -aspii & tiajleii-Dlia-Dispersion

2 Hi332t0n long b © §- "I'JOCO 3 z. S- ^ irri-fiigiext" j £.-Ö -äie tit -erste— - iieB.4e_ Fl ^ asi ^ eit, tlalslie äii J ^ kr-sreserroire contained j wurcle ai3 ^, gjB3 & fi.i5i.3ä »^ iui ^ iSS ji_iiii> -i ^ 'UaI Sa2i-3säti" vits. "fc» 3a d © .s Gliolesteiyioieat;' ± ai-dies-ea ßjfsfc & a a niciit-austaiisclibare

5S

iiS ctarstelli; _s is -las -Fsrjiäl ^ ais from ^ ifc E

-kieiifcetfl ■ Öät-xa-ur.ol '-. ^^ scs ^. * ??? ? '\ ^ Xt- * "3« asxkier-tem

sir * Asissicibn iCi- -Sie Äcgsfcerate (Aasl; raaivIiE-ifcCsniat; s-as äen Siiclit — 7esilralar-

oäss? Te-siltixlaT-ilii ^ oTiSi ^ -rr-i τξγ. ia -you Fäcspfcairicylchclxn— dispersion. jDie Er- ?, 6br..i.ss-3 Eiai i: i öer- Fig. 25 ;, Bar-sss- i £ - "to sr-s *:.: -": that äas üst-radioliaaäecanoat with both forms S- ^ r jülrrorsservoirs asscz & rt islieb, Indicates that there is something for the ir-aasport tmd the Xenobiotics are satisfactory.

The present invention, a xenobiotifcum containing oxide micrreservoir © Tmmieci in the most varied of Bosierungs-frf ^ eu formulated ornaments. You can "for example in a ^" χ "SdIs ^ ^- E io IL-T sis eis" ^Γ 32? Τ] Γ'3-τ1 rl ^ 'i-srn. liquid-free dispersed w €: iea, ice kcEZi-sn troeken s'or- manufacture -yen tablets

over ^ wx-ial best. From a "waisted EescϊlΓeiϊi ^ αιg xuiä dex examples described above, it can be seen daH the invent-ungsgemäße delivery Yehiculum is able to pharmakodynasiischen properties of Xencbiotika with a wide range -.rcn physical chemical nand

909850/0569 BATH ORIGINAL

Properties as well as having a wide range of biological Areas of application and properties to change in an advantageous manner.

It can be seen from the foregoing that the above objects of the present invention are effectively achieved and that the present invention is not limited to the above, but also certain modifications to the implementation of the method described above and to the composition and molded bodies described above, as they are readily apparent to the person skilled in the art.

9098 50/056 9

Blank page

Claims (100)

MIiLLER-IiOItE · I) EUFlIL · MCiIOiV ■ If E WL «" ·, ί'ΛΤ EN TA N Wi L · TE 2015028 C) R. WOUFOANO MÜ LLEf! -BORE (PAiHNTANWALT VON \ 0> I - 197S) E ) F !. PAUL CJElIFEL. DIPL.- CU EM. UH ALFHtC) SCHÖN.OIPL.-CHEM. WEHNHFJ HEEJfKL, [1IPL.-PHY5. L 1193 Arthur D. LittLe, Inc. 2 5 Acorn Park, Cambridge, Massachusetts; 02 140 / USA Delivery Vähiculum for the delivery and release of a xenobiotic within a mammalian host, process for the production of the delivery vehicle and its use in a method for the predetermination and control of the pharmacodynamic properties under which a xenobiotic is delivered or released within a mammalian host Claims 1. Delivery vehicle that is biological with a mammalian host
is compatible, for the delivery and release of a xenobiotic within this host, the pharmacodynamic properties of which are changed in an advantageous manner through the use
this vehicle, characterized in that the delivery vahiculum is in the form of microreservoirs which contain the xenobiotic and contain din or consist of a phospholipid component and one which is immiscible with a phospholipid
Part of lipid which is physiologically tolerable 909850/0569 MC-N (UiKi Uli - -SI I-.U EHTSi'l !. - <· - ΓΙ, .ΛΓ + 'Λ! II'"H). -I '->» liii.f .. .M Ii K-tlli! · I Γ · 'V ii. Lll-ii> ι / Hl ι> · "> · ΐ H I, r. ν S-JUvI ι) υ ) ^! - \ i. ί ι χ y Liquid is stable and therefore essentially immiscible. 2. delivery vehicle according to claim 1, characterized in that that the microreservoirs are in vesicular shape with diameters within of the range between about 190 and about 300 A. 3. delivery vehicle according to claim 1, characterized in that that the microreservoirs in multi-vejicular form with a diameter> srn are within the range between about 250 and about 1000 Å. 4. delivery vehicle according to claim 1, characterized in that that the microreservoirs both in light veöikular form and in Vesicular shape. 5. delivery vehicle according to any one of claims 1 to 4, characterized characterized in that the microreservoirs are about 50 mole ° o to about Contains 97 moles / £ of the phospholipid component. 6. dispensing vahiculum according to any one of claims 1 to 5, characterized characterized in that the microreservoirs contain a xenobiotic binding modifier contain. 7. delivery vehicle according to claim 6, characterized in that the xenobiotic binding modifying agent has taken up of the xonobiotic increased by the microreservoirs. 8. delivery vehicle according to claim 7, characterized in that that the XenobLotikum-BLrulunrjoinodi f izi ^ rungssnl fct-j L is about F'hospha tidsciuru handslt, which has a smaller proportion of the Phosuho LLpLcJ- 9 0 9 8 5 0/0 B B 9 ORIGINAL INSPECTED 291bU28 Constituent forms. 9. delivery vehicle according to claim 6, characterized in that the xenobiotic binding modifier the uptake of the xenobiotic is reduced by the microreservoirs. 10. A delivery vehicle according to claim 9, characterized in that the xenobiotic binding modifier is cholesterol. 11. delivery vehicle according to any one of claims 1 to 10, characterized characterized in that the microreservoirs contain a xenobiotic release rate control agent contain. 12. delivery vehicle according to claim 11, characterized in that that the xenobiotic release rate control agent is miscible with the phospholipid constituent of the microreservoirs Lipid acts. 13. delivery vehicle according to claim 12, characterized in that that the lipid is cholesterol. 14. Release vehicle according to one of claims 1 to 13, characterized in that the phospholipid component is phosphatidylcholine contains or consists of. 15. Delivery vehicle according to one of claims 1 to 14, characterized characterized in that the phospholipid component contains or consists of a mixture of phosphatidylcholine and phosphatidic acid, where the phosphatidic acid is present in the mixture in an amount 9098 5 0/0569 which is at least about 5 molar equivalent. 16. Delivery vehicle according to one of claims 1 to 15, characterized characterized in that the lipid component immiscible with the phospholipid contains or consists of a cholesterol ester a fatty acid with 10 to 18 carbon atoms. 17. A delivery vehicle according to claim 16, characterized in that the cholesterol ester is cholesteryl oleate. 18. Delivery vehicle according to one of claims 1 to 17, characterized characterized in that the lipid constituent which is immiscible with the phospholipid contains or consists of a triglyceride. 19. Delivery vehicle according to claim 18, characterized in that that the triglyceride is glycine trioleate. 20. Delivery vehicle according to one of claims 1 to 19, characterized characterized in that the xenobiotic is a medicament or a drug. 21. Delivery vehicle according to claim 20, characterized in that that the drug is a chemotherapeutic drug. 22. Delivery vehicle according to claim 21, characterized in that the drug is a chemotherapeutic one Anticancer drugs. 9 0 98.5 0 / 0.5 69 ; ^; 29 ΐ 5028 23. Delivery vehicle according to claim 22, characterized in that that the cancer chemotherapeutic agent is a Anthracycline is. 24. Delivery vehicle according to claim 23, characterized in that that the cancer chemotherapeutic agent is adriamycin acts. ■ 25. Delivery vehicle according to claim 23, characterized in that that the cancer chemotherapeutic agent is AD 32, 26. Delivery vehicle according to claim 20, characterized in that that the drug is a parasiticide. 27. Delivery vehicle according to claim 26, characterized in that that the parasiticide is imidocarb. 28. Delivery vehicle according to claim 20, characterized in that that the drug is a fertility control agent acts. 29. Delivery vehicle according to claim 28, characterized in that that the fertility control agent is ostradiol undecanoate acts. 30. Delivery vehicle according to one of claims 1 to 29, characterized in that the xenobiotic is a drug acts, whose plasma kinetics changed in an advantageous manner will. ORIGINAL INSPECTED 909850/0569 291bO28 31 »Delivery vehicle according to one of claims 1 to 30, characterized in that the xenobiotic is a drug whose chemotherapeutic effectiveness is changed in an advantageous manner. 32. Delivery vehicle according to one of claims 1 to 31, characterized in that the xenobiotic is a drug whose toxicity is more advantageous Way is changed. 33. Delivery vehicle according to one of claims 1 to 32, characterized in that the xenobiotic is a drug, its oral absorption and its Ability to pass through the gastrointestinal tract in beneficial Way to be changed. 34. Process for the manufacture of a delivery vehicle, in particular such according to any one of claims 1 to 33, for the delivery of a xenobiotic within a mammalian host, the pharmacodynamic properties in a predictable manner are modified and controlled, characterized in that microreservoirs are made of a composition which contains or consists of a phospholipid component and a lipid component which is immiscible with the phospholipid and which is in one physiologically compatible liquid stable and thus in the essential is immiscible, and that the xenobiotic to be released or released within the microreservoirs incorporated. 909850/0569 35. The method according to claim 34, characterized in that one for the production of the microreservoirs a) with a solvent, a solution of the Phosphalipid component and the lipid component immiscible with the phospholipid manufactures, b) the solvent removed to produce a dry Residue mixture from the phaspholipid component and the lipid constituents that cannot be mixed with the phospholipid, c) the dry residue mixture with a physiologically compatible one Liquid hydrated to make a suspension, d) the suspension in a non-oxidizing atmosphere at a Temperature at least the melting point of the phospholipid The immiscible lipid component is equivalent to being treated with ultrasound (sonicates) to produce the Microreservoirs, and e) separating the microreservoirs thus formed. 36. The method according to claim 35, characterized in that the xenobiotic is wet incorporated into the microreservoirs by adding the xenobiotic to the solution in step (a). 37. The method according to claim 35, characterized in that one the xenobiotic is incorporated into the microreservoirs by adding the xenobiotic prior to performing the ultrasound treatment The suspension of stage (c) is added. 38. The method according to claim 35, characterized in that the microreservoirs formed are separated by the with Ultrasonically treated suspension centrifuged and the during 909850/05 69 Centrifugation obtained clear solution chromatographed, whereby a series of chromatographic fractions is obtained. 39. The method according to claim 38, characterized in that the chromatography fractions which contain the microreservoirs in vesicular form with diameters within the range between approximately 190 and approximately 300 Å are separated from the fractions which the microreservoirs in non-vesicular form with diameters within the range between about 250 and about 1000 Å. 40. The method according to claim 35, characterized in that a xenabiotic binding modifier is added to the solution. 41. The method according to claim 40, characterized in that the xenobiotic binding modifier is the inclusion of the Xenobiotic increased by the microreservoirs. Al. Process according to Claim 41, characterized in that the xenobiotic binding modifier used is phosphatidic acid, which forms a smaller component of the phospholipid component. 43. The method according to claim 40, characterized in that the xenobiotic binding modifier is the inclusion of the Xenobiotic decreases through the microreservoirs. 44. The method according to claim 43, characterized in that the xenobiotic binding modifier is cholesterol used. 9 0 9 8 B 0/0 5 6 9 45. A method according to claim 35, characterized in that the solution added to a xenobiotic-ireisetzungsrate-Kontjjollmittel, 46. The method according to claim 45, characterized in that the xenobiotic release rate control agent is one with the Phospholipid component of the microreservoirs, miscible lipid used. 47. The method according to claim 34, characterized in that the phospholipid component contains or consists of phosphatidylcholine consists. 48. The method according to claim 34, characterized in that the phospholipid component contains or consists of a mixture from phosphatidylcholine and phosphatidic acid, the phosphatidic acid is present in the mixture in an amount that is at least about 5 mole / 5 equivalent. 49. The method according to claim 34, characterized in that contains the lipid constituent that is immiscible with the phospholipid or consists of a cholesterol ester of a fatty acid having 10 to 18 carbon atoms. 50. The method according to claim 49, characterized in that the cholesterol ester is cholesteryl oleate. 51. The method according to claim 34, characterized in that the Contains lipid constituents which are immiscible with the phospholipid or consists of a triglyceride. 909850/0563 52. The method according to claim 51, characterized in that it the triglyceride was glycerol trioleate. 53. The method according to claim 34, characterized in that the xenobiotic is a drug or a drug acts. 54. The method according to claim 51, characterized in that the drug is a chemotherapeutic drug acts. 55. The method according to claim 54, characterized in that the drug is a chemothrapeutic cancer drug acts. 56. The method according to claim 55, characterized in that it the cancer chemotherapy drug is an anthracycline. 57. The method according to claim 55, characterized in that the cancer chemotherapeutic agent is adriamycin, 58. The method according to claim 55, characterized in that the cancer chemotherapeutic agent is AD 32. 59. The method according to claim 53, characterized in that the drug is a parasiticide. 60. The method according to claim 59, characterized in that the parasiticide is imidocarb. 909850/0569
. ORIGINAL INSPECTED - τι - - 61. The method according to claim 53, characterized in that the medicament is a fertility control agent
acts. 62. The method according to claim 61, characterized in that the fertility control agent is oleodiol undecanoate
acts. 63. The method according to claim 34, characterized in that the xenobiotic is a drug whose.
Plasma kinetics is changed in an advantageous manner. 64. The method according to claim 34, characterized in that the xenobiotic is a drug whose
chemotherapeutic effectiveness is changed in an advantageous manner. 65. The method according to claim 34, characterized / that the xeiiobiotic is a drug whose
Toxicity is changed in an advantageous manner. ■ 66. The method according to claim 34, characterized in that the xenobiotic is a drug whose
oral absorption and its ability to pass through the gastrointestinal tract is changed in an advantageous manner. 67. The method according to claim 34, characterized in that one for the production of the microreservoirs 1.) in a water-miscible organic solvent a Solution of the phospholipid component and that with the phospho-Hpid produces immiscible lipid constituents and 909850/0569 2. this solution under conditions under which microreservoirs arise in the physiologically compatible Injected liquid. 68. The method as claimed in claim 67, characterized in that the xenobiotic is incorporated into the microreservoirs adding the xenobiotic to the solution. 69. The method according to claim 67, characterized in that for incorporation of the xenobiotic into the microreservoirs the xenobiotic adds to the physiologically acceptable fluid. 70. The method according to claim 34, characterized in that the microreservoirs containing the xenobiotic are in suspended in the physiologically compatible liquid, thereby converting the xenobiotic into a liquid dosage form. 71. The method according to claim 34, characterized in that the microreservoirs containing the xenobiotic are in one formed from a physiologically compatible material Encapsulated capsule. 72. Method of predetermination and control of pharmacodynamic Properties under which a xenobiotic is released within a mammalian host thereby characterized in that the xenobiotic is released within the host from circulating Mxkroreservoiren, the from a phospholipid component and one with the Phospholipid immiscible lipid constituents are formed in a physiologically acceptable liquid is stable and therefore essentially immiscible. 909850/0569 73. The method according to claim 72, characterized in that
the plasma kinetics of the xenobiotrum includes the pharmacodynamic properties that are predetermined and
to be controlled. 74. The method according to claim 73, characterized in that the xenobiotic is a drug or a Drug acts. 75. The method according to claim 74, characterized in that the medicine is a parasiticide. 76. The method according to claim 75, characterized in that the parasiticide is imidocarb. 77. The method according to claim 72, characterized in that the xenobiotic is a chemotherapeutic drug and that the chemotherapeutic drug
Effectiveness and toxicity of this drug
includes pharmacodynamic properties that are predetermined and controlled. "'■ ■ 78. The method according to claim 77, characterized in that the drug is a chemotherapeutic one Anticancer drugs. 79. The method according to claim 78, characterized in that the cancer chemotherapeutic agent is a
Anthracycline is. 80. The method according to claim 79, characterized in that the cancer chemotherapeutic agent is doxorubicin acts. 9 0 9 8 5 0/0569 14 - 2Ü 150 2 = 8 81. The method of claim 79, wherein the cancer chemotherapeutic agent is AD acts. 82. The method according to claim 72, characterized in that the oral absorption of the xenobiotic includes the pharmacodynamic properties that are predetermined and controlled will. 83. The method according to claim 82, characterized in that the xenobiotic is a fertility control agent acts. 84. The method according to claim 82, characterized in that the fertility control agent is oil stradiol undecanoate. 85. The method according to claim 72, characterized in that the circulating microreservoirs in vesicular form with
Diameters within the range of about 190 to
about 300 A are present. 86. The method according to claim 72, characterized in that the circulating microreservoirs are in non-vesicular shape with diameters within the range of approximately
250 to 1000 8 exist. 87. The method according to claim 72, characterized in that the circulating microreservoirs are both in non-vesicular form as well as in VesikuLar form. 88. The method according to claim 72, characterized in that the circulating microreservoirs are approximately 50 to approximately 97%
Contain MoI- ¹ S of the E'hosphoL ipid component. 909850/0569 ■ - 15 - 2ϋ Ι5028 89. The method according to claim 72, characterized in that that the circulating Mxkroreservoire a Xenobiotic binding modifxzierungsmxttel contain. 90- The method according to claim 89, characterized in that the xenobiotic binding modification mechanism is is phosphatidic acid, which has a smaller proportion of the phospholipid component forms. 91. The method according to claim 89, characterized in that the xenobiotic binding modification mechanism is cholesterol. 92. The method according to claim 72, characterized in that the circulating Mxkroreservoire a xenobiotic release rate control agent contain. 93. The method according to claim 92, characterized in that the xenobiotic release rate control agent is one with the phospholipid component of the Mxkroreservoire miscible lipid. 94. The method according to claim 93, characterized in that the lipid is cholesterol. 95. The method according to claim 72, characterized in that the phospholipid component contains or consists of phosphatidylcholine. 96. The method according to claim 72, characterized in that the phospholipid component contains or consists of a mixture of phosphatidylcholine and phosphatidic acid, the phosphatidic acid in the mixture in an amount is present which is at least about 5 mol-I equivalent. 909850/0569 J SUBMITTED I -ie- 2315Q28 97. The method according to claim 72, characterized in that the lipid constituent which is immiscible with the phospholipid contains or consists of a cholesterol ester of a fatty acid with 10 to 18 carbon atoms. 98. The method according to claim 97, characterized in that the cholesterol ester is cholesteryl oleate
acts. 99. The method according to claim 72, characterized in that the lipid constituent which is immiscible with the phospholipid contains or consists of a triglyceride. 100. The method according to claim 96, characterized in that the triglyceride is glycerol trioleate. 909850/0569