DE10118852A1 - Solid particles for transporting hydrophobic active agents, e.g. drugs or nucleic acids, obtained from organic solvent solution of active agent and water-insoluble and amphiphilic polymers by ultrasonication and dialysis - Google Patents

Abstract

New solid particles (I), for transporting hydrophobic or hydrophobized active agents (A), are obtained by: (a) dissolving (A), a water-insoluble organic polymer (P1), an amphiphilic organic polymer (P2) and optionally supplementary components in organic solvent(s); (b) ultrasonicating the solution; (c) dialyzing the solution against water; and (d) separating the obtained particles (I) from the aqueous solution. Independent claims are included for: (i) new solid particles (I') for transporting (A), comprising a core of (P1) and an outer layer of (P2) (not covalently bonded to the core), where (P2) is polyvinyl alcohol (PVA) or its ester; (ii) the preparation of (I') by the procedure described above for (I); (iii) particles for transporting drugs (A), which (a) have linker molecules (LM), containing amino-reactive and/or thiol-reactive groups, covalently bonded to free amino or thiol groups present on the surface of the particles; or (b) have a mixture of two types of LM's covalently bonded to reactive groups on the particle surface, one type being a bifunctional LM having a further reactive group on the other end of the molecule and the other being a monofunctional LM with no further reactive groups; and (iv) the use of pure bifunctional polyethylene glycol (PEG) molecules (preferably N-hydroxysuccinimide (NHS) ester/vinyl sulfone PEG), optionally mixed with monofunctional PEG molecules (preferably NHS ester-PEG) for the preparation of surface-modified solid particles for transporting drugs.

Description

The invention relates to solid particles for the transport of pharmaceutical Active substances, processes for their preparation, medicaments containing them Particles as well as the use of these particles in various forms chose indications.

A main goal of pharmaceutical research is to find the ones you want Effects of known active ingredients and the systemic side to minimize effects, which is particularly the case for substances with high trinsic and thus inevitable toxicity (e.g. cytostatics) of is of great importance. This can be both about reducing the for the therapeutic effect required total dose as well as accumulation of the effectors can be achieved at the desired site of action, both by the controlled, spatially specific release of effector molecules in the broadest sense (proteins, peptides, nucleic acids or low molecular weight laren substances) in the desired target tissue can. Among them is the specific transfer of therapeutic or diagnostically usable substances in defined biological targets ("drug delivery", "drug targeting") to understand which is an important target of current pharmaceutical research is.

The antibody technology available today allows the generation of high affinity binding partners for almost any biological structure door; moreover, there are numerous natural ligands for cellular receptors  have been characterized and cloned so that it is no longer a problem Molecules with high and specific affinity for the desired targets to create. In many cases, low molecular weight ligands (e.g. Glycosides) known, which imitates chemically and thus for Target control can be used. However, these "finder moles practice cool ", whether micro- or macromolecular, generally no phar itself pharmaceutically usable function, while the effectors themselves are not are target-specific. A main focus must therefore be on this Bridge separation and the therapeutic potential of the available To connect effectors with the target specificity of the searcher molecules.

The most efficient approach known to date for achieving this goal is to use support structures in the colloidal (ie sub-micrometer) size range, on the surfaces of which corresponding target search molecules can be bound. In this way, both the optimal ratio of homing to effector molecules and maximum flexibility are achieved: While only a few (<10) effector molecules can be bound by direct covalent coupling to a single antibody or other ligand molecule (which is the case with a molecular weight of an antibody) of around 150 kDa means that over 90% of the mass of the conjugate is accounted for by the antibody part) and conjugates with low molecular weight finders usually have a molar ratio of 1: 1 is an effector with colloidal systems: target seeker ratio of 10 ³ -10 ⁴ realizable. In addition, no chemical change to the effector is required, which is advantageous in every respect.

An efficient way to do this is to use the relevant substance zen in colloidal carrier particles, which ge with antibodies genes or natural ligands for characteristic molecular structures linked to the goal and at the same time by inert coating of their surface be protected against the immune system.

A widely used method for colloidal packaging of pharmaceuticals is to enclose the effectors in lipid membrane-enveloped vesicles (liposomes). By using appropriate membrane components, it is possible, on the one hand, to bind the desired targeting molecules to the liposome membranes, on the other hand to coat the carriers with anti-immunogenic coatings (e.g. polyethylene glycol) and thus prior to the non-specific removal from the blood stream through the reticuloendothelial system to protect. The advantages of this system are offset by the following serious disadvantages:

• - The thermal and temporal stability of a single lipid dop The existing vesicle layer is limited, as is the tightness. The Permeability of the membranes for hydrophilic substances can in principle ver be reduced, but the required changed (e.g. fluorinated) Lipids not biologically harmless. Moreover, a single one is enough "Hit" of the complement system to expire a complete Ve sikels.  
• - The lack of stability of the membrane vesicles in turn limits the possible variability in the surface design and thereby limited the potential applications.
• - Only hydrophilic substances can in the aqueous inner phase of the Ve Sicles are transported in sufficient concentration.
• - The liposomes are loaded (read from a few special cases hen) by simple inclusion of part of the aqueous phase and is accordingly inefficient: Typically <0.5% becomes the effector substance enclosed in the vesicles. Here, the substance be exposed to severe thermal and chemical loads (the Ar Working temperature must be above the critical phases for a long time transition temperature of the lipid mixture, and that for the covalent Mo reactive groups required for this only survive when they are very low pH value).
• - Half of the membrane-related reactive groups that link test with proteinaceous search molecules is located after the Vesicle formation on the inside and is not available for binding supply, but is released after the liposomes have dissolved in the organism set and can lead to unpredictable reactions.
• - The chemical coupling of proteins to liposomal membranes (e.g. via lipid linked directly or via a polyethylene glycol arm SPDP) leads to the formation of highly immunogenic structures  Vaccination have been used successfully in the field of "drug targeting ", however, must be viewed as unusable because they are too cause an immune reaction against the particles.

Solid colloidal particles ("nanoparticles") are available as an alternative supply. In principle, nanoparticles are known. Particles in the micrometer and In principle, submicrometer range made of hydrophobic polymers can by fine dispersion of the in a non-polar solvent taken polymer: By removing the solution means the polymer falls into the form of particles, the diameter of which is below from which the droplet lies; loading with hydrophobic substances (in which category most drugs fall) by simple Addition of the substance to the non-polar solvent can be accomplished: After removing the solvent, the active substance is too close 100% associated with the polymer before and remains when the particles are in a aqueous phase are introduced by Van der Waals forces and steri "Entrapment" not covalent, but stable in the long term bound kelmatrix. It is essential here that a subsequent coagula tion of the hydrophobic particles (their large contact area with the hydrophilic medium is energetically unfavorable) is prevented. At off conventional approaches known in the art this is usually done by producing the particles in the presence of an amphiphi len substance, which between hydrophobic particle matrix and hydrophi conveyed to the medium.  

Conventional nanoparticles and their production and possibilities a surface variation are known from WO 96/20698, the System here especially for intravascular use, especially for the restenosis was optimized. The patent application describes polymo molecular nanoparticles made from natural or synthetic polycondensates with a predominantly generally described property modification surface coating made of natural or synthetic macromo read.

The following documents may also be mentioned as examples.

DE 198 10 965 A1 describes polymolecular nanoparticles from one Polyelectrolyte complex of polycations and polyanions, which with a Crosslinking agent is treated.

No. 6,117,454 describes in particular polymolecular nanoparticles, by a coating of fatty acid derivatives to penetrate the Blood-brain barrier are suitable. In these inherently amphiphilic Li ganden is to be considered that on the one hand high affinity ligands i. w. S. against biological structures (e.g. proteins) i. a. not available on a scale are bar that they themselves in the presence of appropriate physi properties (which, for example, does not apply to antibodies) for immediate suitable surface coating (for which ver with the particle matrix comparable quantities are required), on the other hand even where this is the case is possible from introducing such masses with high affinity for biological targets of binding molecules, which are expected to separate  partially detach from the particles and regardless of their goals bind, is strongly advised against.

In US 5,840,674, covalently bonded via a "linker" Complexes of active ingredient and microparticles, in particular for use described microorganisms.

No. 5,641,515 describes polymolecular nanoparticles made of polycya nacrylate containing insulin that controls the complex bound insulin release.

DE 198 39 515 A1 describes colloidal polymer-active substance associates with a property-optimized branched polyol ester especially described on mucosal tissues. Thereby a polyme res polyol, especially polyvinyl alcohol, with, for example, polycondensation Hydroxy carboxylic acid esterified so that starting from the polyol Backbone polycondensed side chains of different lengths and one terminal free OH group arise with the aim of the properties to change the polyol ester. The fact that the side chains of the after Particles produced in DE 198 39 515 A1 exclusively in free OH End groups is very disadvantageous, since OH groups in aqueous or al alcoholic milieu can not be implemented selectively, so that another Surface modification, for example, with "finder" molecules rig or almost excluded. Next is for particle formation according to DE 198 39 515 a dispersion step in an aqueous phase is absolutely necessary, so that particles produced by this process cannot be modified further  are. In particular, however, the particles according to DE 198 39 515 also not have a sufficiently controlled, clearly defined structure, because No molecules are added that define the length of the pages chains are to be controlled by a chain termination reaction and also none for further modification favorable groups in the surface of the molecule be introduced. The manufacturing process of the polymer u. a. because of the terminal OH groups much more complex and (as a result the influence of numerous process parameters) less robust and leads also to a less structured polymer, which is not self-evident is capable of forming monomolecular particles, but "by con trolled precipitation into colloidal form ". The resulting "Colloidal Associates" are lacking due to the lack of defined ones End pieces of chemically different from the rest of the particle, such as probably covalently associated surface layer with which one defined surface modification can be carried out. A total of the particles according to DE 198 39 515 are therefore suitable for use the field of "drug targeting" in particular through surface modification cation in the sense of this invention are unsuitable and are only as "sustained release" formulations applicable.

In the prior art, the problem of targeted Ap application of the drug at the desired site and the stability of the Nanoparticles after administration until they reach the site of action, the "Drug Tar geting ", only insufficiently solved. A central problem is the O surface modification of nanoparticles. As mentioned above,  represent conventional solid nanoparticulate systems (in contrast to Liposomes) due to their extremely high specific interphase between hydrophobic particle matrix and hydrophilic medium an energetically un cheap system that is unstable in the absence of stabilizers. By storing particles together, the energetically unfavorable Minimized contact area, precipitation of the hydrophobic Particle material is the result.

For this reason, conventional nanoparticles need a coating for example, from amphiphilic molecules that make up the interfacial energy lower and stabilize the particles in this way. This coating completely covers the particle matrix and thus deprives it of access modifying agents. A modification of the amphiphilic mo On the other hand, leküle leads to a drastic change in the physi calic properties and thus destabilizing the coating. For this reason, it is hardly possible to use conventional nanoparticles in this way modify that binding of ligands and thus use in the field of "drug targeting" is possible, which, as already stated, also applies to the DE 198 39 515 applies.

In addition to overcoming the aforementioned disadvantages of the prior art, it was an object of the invention to provide nanoparticles that

• a) a wide range of active ingredients through "steric entrapment" can transport  
• b) surface modification can be carried out simply and stably, and in the process a total of
• c) Show defined and suitable chemical groups on the surface or can be easily manufactured with it and
• d) a defined size and in particular the shape and surface of the mo show leküls

and in particular are able to release a specific we to reach kort.

The solution to this problem is achieved by solid particles for transporting hydrophobic pharmaceutical active ingredients

• a) an unbranched or a maximum of three branched molecule back burr from a polymer made up of monomers with at least a binding group (x) on each monomer, with the bin Formation groups (x) each covalently via a (x) - (x ') bond
• b) polycondensed molecular side chains constructed from chain formers the monomers each having at least one binding group (y) and at least one binding group (x ') or built up from ver different chain-forming monomers, of which one monomer min at least two binding groups (y) and another monomer min at least two binding groups (x '), or built up from various chain-forming monomers, one of which is a monomer  at least two binding groups (y), another monomer min at least two binding groups (x ') and another at least one Binding group (y) and at least one binding group (x ') points, where (x ') a covalent bond with (x) and also a ko valent bond with (y) can bind, with each on End of the molecular side chains covalently via a (y) - (y ') bond
• c) side chain end pieces which have at least one binding group (y '), no binding group (y) and at least one optionally with a free group (z) provided with a protective group are,

in which

• - The molar ratio between the monomers of the molecular side chain ten (b) and the side chain end pieces (c) << 1,
• - the group y ≠ of group z and the group x '≠ of group z,
• - The molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
• - x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y or y / y 'corresponding pairs of bonds x / x' , x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -OC (O) -), NH ₂ / COOH (amide bond -NH-C (O) -), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH-C (O) -) or COOH / SH (thio-ester bond -S- C (O) -) and
• z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group,

contain.

Protecting groups for certain functional residues and their removal are well known to the person skilled in the art. Possible protective groups would be for example FMOC to protect an amino function, the Removal by treatment with catalytic amounts of piperidine follows.

The particles according to the invention are particularly suitable forms with de enhancing the effects and minimizing the side effects through the controlled and / or spatially specific release of the Ef fector molecule is reached. Generally, it is from the Er particles encompassed preferably around solid, colloidal and / or lipid-free particle systems.

A particular advantage of the groups according to the invention is that due to the presence of functional groups, in particular amino groups, which are not otherwise found in the polymer particle, the surface layer is formed in such a way that

• - The particles with one by the charge of these functional groups conditioned polar zone, the charge of which is encased Flocculation counteracts and thereby the particle suspension stabilized, and
• - functional groups (especially, but not necessarily, from finally, amino groups) used to attach upper surface modifications after the formation of the substance of Particles loaded with interest are suitable to be provided.

Furthermore, functional groups on the surface are targeted depending on the task be easily selectable via the side chain end pieces and the manufacturer lungsverfahren for the compounds of the invention is simple and robust.

In addition, the size, shape and surface of the molecule are controllably clear defined because the choice of molar ratios allows interventions here.

Furthermore, the particles are distinguished by the polymeric side chains in able to use "steric entrapment" large amounts of different To bind and to actives of different chemical properties transport.

The invention also relates to solid particles for Transport of amphiphilic or lipophilic active substances or hydrophobic Re sorption ester hydrophilic active ingredients containing a molecule of a ver branched polycondensate, the polycondensate from a backbone from a multifunctional, preferably unbranched or little  branched, macromolecule, preferably polyvinyl alcohol, exists functional groups (in the case of polyvinyl alcohol, the OH groups) with secondary, preferably also unbranched or little ver branches, polycondensate chains are covalently linked, which in fol secondary polycondensate chains referred to as side chains linked bifunctional organic monomers (where the monomers are concerned with heterobifunctional molecules or their derivatives can or an equimolar mixture of two homobifunctional Molecules or their derivatives), preferably from esterified hydroxycarbon acids or a combination of diols and dicarboxylic acids in the molar Ratio 1: 1 exist, the end of each of these chains from an im Meaning of the condensation reaction monofunctional molecule, preferred a non-hydroxycarboxylic acid.

In Fig. 2a the formation and structure of a general polycondensate from an unbranched, multifunctional backbone, a heterobifunctional side chain monomer and an end piece matching this combination is shown schematically. Of course, the combination of several types of monomers and / or end pieces in a single synthesis reaction is also possible. Such a polycondensate with an unbranched backbone and unbranched or little branched side chains is referred to below as ktenate due to its shape (gr. Kteés = comb).

Here, the original can be used in the formation of the side chains Monomers are used, which in the condensation reaction What split off, or their intramolecular anhydrides, lactones etc. or on their derivatives. A use of "prefabricated" oligomers with im In terms of the condensation reaction, freely available functional groups (e.g. oligopeptides) alone or in any combination with homo- or heterobifunctional monomers is also possible; such before Manufactured blocks are also called below "Monomers" are subsumed.

In the following, some examples of are not exhaustive Substances and combinations which can be used according to the invention listed:

backbone

Side chain monomers

Tails

The selection of the end pieces depends on the following factors:
Desired exposed group (s) (this must be provided with a suitable protective group before synthesis in order to prevent inclusion in the polycondensation process),
Side chain monomer (s),
Backbone molecule (defines the orientation of the side chains through its functional groups).

These side chains are generally referred to as telo-end-piece poly-monomers (i.e. telo-alanyl polylactide or equivalent), the whole molecule therefore as a backbone _{molecular weight} -telo {free group} end-molecule therefore as a backbone _{molecular weight} -telo {free group} end-piece- Poly-monomer _{medium side chain} weight -at, so z. B. Polyvinyl 200,000 telo {amino} alanyl polylactide ₅₀₀₀ at, Polyacryl 50,000 telo {amino, sulfhydro} cysteyl poly (glycol: adipine) 8000at (or accordingly).

An overview of some selected options is given in the following table, which also suggests trivial names for some of the most interesting basic structures:

All of these possibilities are independent of the type and orientation of the binding structure dominating the particle matrix Invention since they are all in the same elongated, through covalent Bonds held together particle shape and structure with hydro phobic core and functionalized, hydrophilic outer layer result, which hereinafter referred to as "Bdellosom" (Greek bdella = leech) and meets the requirements described at the beginning.

Therefore, only the synthesis and use of regular lactic acid alanine ktenate (exact name according to the above Nomenclature: Polyvinyl-telo {amino} alanyl-polylactidate) represented because the other basic structures are not too different Particles.

The polymers described in the list have different ones Suitability depending on objectives; so z. B. amino-containing, but thi oil-free finder molecules with little variation of the protocol (first Linking viewfinder and linker, then reaction of the particles with the Viewfinder-linker complex) by means of the usual, but here reversely oriented NHS ester PEG vinyl sulfone linker to particles  from thioctenate, inverse thioctenate or thioamidoctenate will.

It is particularly preferred if the particles according to the invention noncovalently bound, hydrophobic or hydrophobized contain active pharmaceutical ingredient.

Hydrophobized active substances are originally meant more hydrophilic active ingredients that are more hydrophobic through chemical modification became. One example is hydrophobic resorption esters hydrophi Active ingredients.

Another preferred object of the invention, which also achieves the object and has the preferred properties mentioned, are monomolecular solid particles for transporting hydrophobic or hydrophobized active ingredients which can be prepared by a process in which unbranched or maximally three-branched polymer backbone composed of Mo nomers with at least one binding group (x) on each monomer

• a) chain-forming side chain monomers, each with at least a binding group (y) and at least one binding group (x '), where (x') is both a covalent bond with (x) and with (y) can die,
• b) a mixture of chain-forming side chain monomers, where of at least one monomer at least two binding groups (y)  and at least one other monomer has at least two bonds has groups (x '), where (x') is both a covalent bond can enter into with (x) as well as with (y), or
• c) a mixture of chain-forming side chain monomers, where of at least one monomer at least two binding groups (y), at least one other monomer, at least two bond groups (x ') and at least one further monomer at least one Binding group (y) and at least one binding group (x ') has, where (x ') both a covalent bond with (x) and can come in with (y)

and at least one side chain end piece, with at least one binding group (y '), without binding group (y) and with at least one free group (z), optionally provided with a protective group, where (y') can form a covalent bond with (y) ,
is brought into contact under conditions which allow a bond between the monomers or monomer mixtures of the side chains and the polymer backbone and the chain end pieces and also a polycondensation of the monomers or monomer mixtures of the side chains,
in which

• - The molar ratio between the monomers of the molecular side chain ten (b) and the side chain end pieces (c) << 1,  
• - the group y ≠ of group z and the group x '≠ of group z,
• - The molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
• - x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y or y / y 'corresponding pairs of bonds x / x' , x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -OC (O) -), NH ₂ / COOH (amide bond -NH-C (O) -), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH-C (O) -) or COOH / SH (thio-ester bond -S- C (O) -) form,
• - z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group and
• - The side chain monomers as pure monomers or derivatives as in tramolecular anhydrides or lactones can be used, so long with them and / or other side chain monomers Ket can form.

Another preferred object of the invention, which also solves the problem and has the preferred properties mentioned, are monomolecular solid particles for the transport of hydrophobic or hydrophobized active substances which can be prepared by a process in which unbranched or maximally three-branched polymer backbone composed of monomers with with at least one binding group (x) on each monomer
polycondensed molecular side chains composed of monomers each with at least one binding group (y) and at least one binding group (x ') or composed of different monomers, of which one monomer has at least two binding groups (y) and another monomer has at least two binding groups (x'), or composed of different monomers, one monomer having at least two binding groups (y), another monomer having at least two binding groups (x ') and a further at least one binding group (y) and at least one binding group (x'), where (x ') a covalent bond can form both with (x) and with (y), whereby at the end of the molecular side chains in each case covalently via a (y) - (y ') bond, side chain end pieces which have at least one binding group (y'), no binding group ( y) and have at least one free group (z) optionally provided with a protective group, where (y ') can form a covalent bond with (y),
is brought into contact under conditions which allow a bond between the polycondensed molecular side chains and the polymer,
in which
the molar ratio between the monomers of the molecular side chains (b) and the side chain end pieces (c) << 1,
is the group y ≠ of the group z and the group x '≠ of the group z,
the molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y and y / y 'respectively. x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -OC (O) -), NH ₂ / COOH (amide bond -NH-C (O) - ), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH- C (O) - ) or COOH / SH (thioester bond -SC (O) -) and
z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group.

For example, the free OH groups of the polyvinyl alcohol hol backbone with the polycondensate-forming hydroxycarboxylic acid esterified, while the non-hydroxy carboxylic acid as the end of this kur zen side chains. The function is the non-hydroxycarbon acid that of the radical scavenger in radical polymerization reaction NEN analog.  

It is preferred that the contacting is carried out in an anhydrous one organic solvent, preferably pyridine, taking place there is also inexpensive and is a preferred embodiment of the method, when contacting takes place in the presence of thionyl chloride and / or - if necessary - for example after the implementation with Protective groups can be split off from thionyl chloride if necessary. Protecting groups for certain functional residues and their removal are well known to the person skilled in the art. Possible protective groups would be for example FMOC to protect an amino function, the distance tion by treatment with catalytic amounts of piperidine.

Usually either the deprotection closes or - if this is not necessary - directly after the implementation, for example Thionyl chloride a wash, preferably with dichloromethane, however naturally also with other water-free - mostly non-polar - organic ones Solvents.

The resulting particles are used to transport the load the hydrophobic active pharmaceutical ingredient. Therefore closes then a further process step in which the product is subsequently together with the hydrophobic or hydrophobic to be transported based pharmaceutical active ingredient in an anhydrous organic solvent Solvent is then dissolved, the solution for some time, preferably over Night, preferably at room temperature, then the solution is saturated with water and then the water-saturated solution in  a larger volume of water is dissolved, if necessary mechanical treatment (preferably not by Ultra sound) and then, if necessary, the particles are cleaned and isolated.

Such treatment is usually not necessary, and it is within limits This invention also preferred if after the solution of the water saturated solution in a larger volume of water no mechanical Treatment follows.

For the final cleaning (and insulation) is preferably - so if this is necessary at this stage of the procedure - towards dialysis resorted to.

When choosing the solvent for the step for loading with active substance, it is preferred if the anhydrous organic solvent in water in the ratio solvent: water between 1:10 and 1:50, preferably between 1:20 and 1:40, in particular between 1:20 and 1:30 solves, and / or is preferably selected from:
Methylene chloride or benzyl alcohol, preferably benzyl alcohol.

The selection does not have to be limited to this, as long as a certain one a defined degree of water solubility is given.

It is particularly preferred if that in the particles according to the invention Polymer backbone unbranched or branched at most once, preferred is unbranched.  

The following conditions alternatively apply to particularly preferred embodiments of the particles according to the invention individually, partially or altogether, namely:
the monomers of the side chain each have a maximum of two groups (y) and a maximum of two groups (x ') and / or
the group (y) in the monomers of the side chain corresponds to the group (x) in the polymer backbone and / or
the group (x ') in the monomers of the side chain corresponds to the group (y') in the side chain end piece and / or
the group (z) is selected from the "free groups" OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group,
the monomers of the side chain each have a maximum of 2 to 10 carbon atoms, preferably 2 to 6 carbon atoms, in particular 2 to 4 carbon atoms, and / or
the monomers of the side chain which have both the group (y) and the group (x '), either only 1 group (y) and 1-2, preferably 1, groups (x') or only 1 group (x ') and 1-2, preferably 1, groups (y) and / or
the monomers of the side chains are identical except for the side chain end piece with 1 group (y) and 1 group (x ') or the monomers of the side chains are constructed identically monotonously alternating except for the side chain end piece from an alternating monomer with 2 groups (x') and a monomer with 2 groups (y).

Among other things, the term "free groups" as a definition for the group (z) as selected from OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group, is a fixed definition in the sense of this Invention.

In a further preferred embodiment of the particles according to the invention, the polymer backbone is selected from
Polyvinyl alcohol, polyacrylic acid, polyvinylamine, polysaccharide or polyamino acid,
preferably polyvinyl alcohol or polyacrylic acid,
especially polyvinyl alcohol.

In a further preferred embodiment of the particles according to the invention, the monomers of the side chain are selected from
Hydroxycarboxylic acids, amino acids, the combination of diamines and dicarboxylic acids or the combination of diols and dicarboxylic acids, or their derivatives,
preferably hydroxycarboxylic acids or the combination of diols and dicarboxylic acids or their derivatives,
in particular hydroxycarboxylic acids such as lactic acid, glycolic acid, tartaric acid or citric acid or their derivatives.

In a further preferred embodiment of the particles according to the invention, the side chain end pieces are selected from
unprotected amino acids, N-protected amino acids, COOH-protected amino acids, unprotected amino alcohols, N-protected amino alcohols, O-protected amino alcohols, unprotected thiol alcohols, O-protected thio alcohols, S-protected thio alcohols or unprotected thiol acids, S-protected thiol acids, CO-protected Thiol acids, unprotected thioamines, S-protected thioamines or N-protected thioamines,
preferably
unprotected amino acids, N-protected amino acids, unprotected amino alcohols, N-protected amino alcohols, unprotected thiol alcohols, S-protected thio alcohols or unprotected thiol acids, S-protected thiol acids,
in particular
unprotected amino acids, such as alanine, N-protected amino acids, such as N-FMOC-β-alanine, unprotected thiol acids or S-protected thiol acids.

"A-protected" in the sense of this invention means that a functional group "A" is provided with a protective group.

In a particularly selected preferred embodiment of the particles according to the invention, the polymer backbone, the monomers of the side chain or their derivatives and the side chain end piece or its derivatives are selected from one of the following combinations:

preferably

in particular

It is one for all of the inventive particles described above preferred embodiment of this invention when the particles are nano are particles and correspondingly a length <5 µm, preferably <3 µm, in particular <2 µm and / or a thickness and width of <200 nm preferably have <75 nm, in particular <30 nm.  

Nanoparticles are particles with a size of less than 1 µm in at least two dimensions. In particular, nanoparticles have a volume below 1 µm ³ . Nanoparticles are solid colloidal particles.

In a particularly preferred embodiment of the invention, the particles according to the invention are specially surface-modified. It is precisely these particles that achieve the object of the invention in an outstanding manner, since they are particularly suitable for targeted transport. The invention therefore furthermore relates to particles according to the invention for the transport of active pharmaceutical ingredients to the linker molecules which have a reactive group (z ') selected from groups which are free from groups which are selected from the groups (z) defined above Groups "(z) selected from OH, SH, COOH or NH ₂ and the vinyl or epoxy group can form a covalent bond, preferably have an amino- or thiol-reactive group, in particular an amino-reactive group, covalently via (z ' ) - (z) bonds with groups (z) present on the surface of the particle selected from the "free groups". In the case of the particles according to the invention, these “free groups” (z) are made available on the surface (optionally after removal of the protective group) by the side chain end pieces.

Linker molecules are understood to be polymers, especially un branched polymers, the properties, especially the surface properties, of the particle, but especially to steric  favorable connection of other bioactive compounds to the parties Serve kel or, if necessary, sterically protect the particles from degradation Zen.

Under "reactive group" [(z ') as well as other (z ")] are in particular to understand groups known in the art that are slightly covalent to the "free groups" (z), especially amino, Bind thiol, carboxy or hydroxy groups, as well as to epoxy or Vi nyl groups.

It is particularly preferred if the linker molecules are bifunctional and, in addition to the reactive group (z ') which binds to the particle according to the invention, also a further reactive group (z ") selected from reactive groups at another end of the molecule can form a covalent bond with one of the groups (z) selected from the "free groups" (z) selected from OH, SH, COOH or NH ₂ and the vinyl or epoxy group, preferably a thiol-reactive group, where z '≠ z ".

It is also a preferred embodiment of the invention if for example the thiol-reactive group on the particle according to the invention binds and (for bifunctional linkers) at another end of the line ker molecule would be an amino-reactive group, for example. The vote The reactive groups of the linker molecules depend on the one hand on the or the free group (s), preferably the free group, on which inventions particles according to the invention to which the linker binds. On the other hand, it hangs  of a further modification, if any, or that made by the linker transferred surface property, in particular a possible second reactive group on the linker molecule of the nature of a possibly still moles to be synthesized or already synthesized on the linker determined.

It is also preferred if the the linker molecules are a mixture of the described bifunctional molecules and monofunctional molecules, which in addition to the reactive to the particle of the invention Group (z '), preferably the amino- or thiol-reactive group, in particular especially the amino reactive group, at no other end of the mole küls another, differently reactive functional group (z ") with z '≠ z" have, are.

These particles are very cheap because bulky residues like antibodies attach to them the bifunctional residues can be attached unhindered The monofunctional linkers sterically prevent degradation. These Particles are also known as acanthus.

It is again particularly preferred if this is on the surface Particles (acanthospheres) significantly more, preferably at least 100% more, monofunctional than bifunctional molecules covalently bound are.

It is preferred if bioactive on the bifunctional linker molecules Macromolecules or "finder" molecules selected from peptides, proteins;  preferably antibodies, antibody fragments or Antibody derivatives with target-binding properties such as "single chain" - Antibodies; Hormones, sugars, preferably glycosides; synthetic or natural receptor ligands; Proteins or Peptides with a free cysteine group or thio sugars, over one Binding to the reactive group (z ") are coupled, are coupled or were coupled before the surface modification.

Under the term "finder" molecule in the sense of this invention can generally be coupled to the particles according to the invention Compounds that are capable of affinity to the biological targets of the active substances, as if there were proteins, peptides, Polysaccharides, oligosaccharides, lipoproteins, glycoproteins or others biological molecules either in healthy tissue (physiological) or expressed (pathologically) in or near diseased tissue tie. "Finder" molecules can, for example, peptides, proteins, for example antibodies, antibody fragments or antibody derivatives with target-binding properties such as "single-chain" antibodies; Hormones, sugars, for example glycosides; synthetic or natural Be receptor ligands. Antibodies, derivatives, are particularly preferred. fragments and glycosides.

It is further preferred if attached to the bifunctional linker molecules bioactive macromolecules or more generally "seeker" molecules, preferably antibodies, antibody fragments or antibody derivatives with target-binding properties such as B. "single-chain" antibodies,  the properties such as B. "Single-chain" antibody, especially with free cysteine group, via a bond to the reactive group (z ") are coupled, are coupled or before the surface modification were coupled. This applies in particular to particles whose coating tion contains significantly more monofunctional than bifunctional molecules. Regardless of the order of reactions, this is a Obtained fully covalently linked macromolecule, the following Architecture features: a central axis made of polyvinyl alcohol, from which sen OH groups hydrophobic polycondensates from hydroxycarboxylic acids go out, which conclude with non-hydroxycarboxylic acids, which either which end freely in hydrophilic groups or those with polymeric, hydrophilic Ren linker are linked, with part of this linker again to connect "finder" molecules.

It is also preferred if the bifunctional linker molecules are bio active micromolecules or "seeker" molecules, preferably sugar, especially thio sugar, hormones or proteins, especially those with free Cysteine group, coupled via a bond to the reactive group (z ") are coupled or coupled before the surface modification pelt. This applies in particular to particles whose coating is above consists predominantly or entirely of bifunctional molecules.

It is further preferred if the particles according to the invention follow Binding of the bioactive micromolecules or "finder" molecules still free reactive groups (z ") are saturated, preferably with cysteine.  

It is another preferred embodiment of the surface modification graced particles when the linker molecules were monofunctional molecules are, in addition to the reactive to the particle of the invention Group (z '), preferably the amino- or thiol-reactive group, in particular especially the amino reactive group at no other end of the molecule have another, differently reactive group (z ") with z '≠ z".

It is particularly preferred if the linker molecules are polyglycolides are preferably polyethylene glycol derivatives, especially NHS ester Polyethylene glycol or NHS ester / vinyl sulfone polyethylene glycol.

In general, any subsequent cleaning or insulation is carried out tion preferably via dialysis, preferably with selective Exclusion membranes.

For all particles according to the invention it is preferred if the too trans porting active pharmaceutical ingredient a synthetic or natural Active ingredient, a protein, peptide, lipid, sugar or nucleic acid or a low molecular organic or high molecular organic activity substance, for example a hormone, an antineoplastic substance, an An tibiotic, antifungal, parasite, virustatic or antihelminti kum, a cardiovascular active substance; a central active substance, in particular an analgesic, antidepressant or antiepileptic; is.

In general, it is a preferred embodiment of the particles according to the invention if the particle according to the invention is linked directly or via a linker, preferably via bifunctional polyethylene glycol molecules, to a "finder" molecule selected from:
Peptides, proteins; preferably antibodies, antibody fragments or the antibody derivatives with target-binding properties such as "single-chain"antibodies; Hormones, sugars, preferably glycosides; synthetic or natural receptor ligands; Proteins or peptides with a free cysteine group or thio sugars.

The processes for producing particles according to the invention are also an important part of the invention. Another object of the invention is therefore a method for producing a particle according to the invention, in which an unbranched or at most three-branched polymer backbone composed of monomers with at least one binding group (x) on each monomer
Monomers each with at least one binding group (y) and at least one binding group (x '), where (x') can form a covalent bond with both (x) and (y),
a mixture of monomers, of which at least one monomer has at least two binding groups (y) and at least one other monomer has at least two binding groups (x '), where (x') forms a covalent bond with both (x) and (y) can, or
a mixture of monomers, of which at least one monomer has at least two binding groups (y), at least one other monomer has at least two binding groups (x ') and at least one further monomer has at least one binding group (y) and at least one binding group (x'), where (x ') can form a covalent bond with both (x) and (y),
and at least one side chain end piece, with at least one binding group (y '), without binding group (y) and with at least one free group (z), optionally provided with a protective group,
in which
the molar ratio between the monomers of the molecular side chains (b) and the side chain end pieces (c) << 1,
is the group y ≠ of the group z and the group x '≠ of the group z,
the molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y and y / y 'respectively. x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -OC (O) -), NH ₂ / COOH (amide bond -NH-C (O) - ), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH- C (O) - ) or COOH / SH (thio-ester bond -SC (O) -) form,
z is selected from CH3, OH, SH, COOH or NH2 and the vinyl or the epoxy group, optionally protected by a protective group and
the side chain monomers can be used as pure monomers or derivatives such as intra-molecular anhydrides or lactones, as long as they can still form chains with themselves and / or other side chain monomers,
is brought into contact under conditions which allow a bond between the monomers or monomer mixtures of the side chain and the polymer backbone and the side chain end pieces and also a polycondensation of the monomers or monomer mixtures of the side chain,
optionally in an anhydrous organic solvent, preferably pyridine, optionally in the presence of thionyl chloride and then optionally purified, preferably by dialysis against H ₂ O, and optionally isolated,
and then optionally the particles with which the hydrophobic or hydrophobized active ingredient is to be transported are dissolved in an anhydrous organic solvent, then the solution is incubated for some time, preferably overnight, preferably at room temperature, then the solution is saturated with water and then the water-saturated Solution is dissolved in a larger volume of water, optionally followed by mechanical treatment and then optionally cleaned the particles, preferably by dialysis against H ₂ O, and isolated.

Another object of the invention is a method for manufacturing of a particle according to the invention (with linker molecule) in which particle according to the invention (without linker molecule), which has a group (z) selected from the free groups containing a linker molecule a reactive group (z ') with the group (z) a covalent bond can enter, includes, to form this covalent bond suitable conditions and then connect if necessary ßend cleans the particles, preferably by dialysis against WO, and iso liert. For example, neutral to weakly basic conditions are suitable gene.

Another object of the invention is a method for producing a particle according to the invention (with linker molecule and "finder" molecule), in which, following the above process, particles (with linker molecule) produced thereafter, which are attached to bifunctional linker Molecules have a free reactive group (z ") with bioactive macromolecules or" finder "molecules as defined above to form a bond between the group (z") and the bioactive macromolecules or "finder" molecules under suitable conditions in Brings contact and optionally then cleans the particles, preferably by dialysis against H ₂ O, and isolated.

The particles according to the invention are particularly suitable forms for Strengthening the desired effects of known active ingredients and to mini mation of systemic side effects through controlled and / or spatially specific release of the effector molecule is achieved. There they are suitable and intended for therapeutic agents of various kinds to be used. Another object of the invention are therefore Medicaments, the particles according to the invention and, if appropriate, suitable Contain nete additives and / or auxiliaries.

In principle, the pharmaceuticals according to the invention can be used as liquid neiforms in the form of aerosols, injection solutions, drops or juices or as semi-solid pharmaceutical forms in the form of granules, tablets, pellets or capsules.

Suitable additives and / or auxiliary substances are e.g. B. solution or diluent agents, stabilizers, suspending agents, buffer substances, con Serving agents, as well as dyes, fillers, and / or binders. The Selection of the auxiliary substances and the amounts to be used depends on them depend on whether the drug z. B. inhalative, oral, oral, parenteral, intra vasally, intravenously, intraperitoneally, rectally, subcutaneously or intramuscularly should be applied. Preparations are suitable for oral applications in the form of tablets, dragées, capsules, granules or suspensions such as drops, juices and syrups, suspensions for other applications as well as easily reconstitutable dry preparations.  

The particles according to the invention are also particularly useful as diagnostics is suitable because, for example, they specifically insert markers into the correct cell can. Another object of the invention is therefore a diagnosis cum, the particle according to the invention and, if appropriate, suitable additives contains constituents and / or auxiliary substances.

Furthermore, as stated above, the particles according to the invention are particularly suitable forms with which to increase the effect and Minimization of side effects through controlled and / or space Lich specific release of the effector molecule is achieved, so that ei ne general usability of these particles for the production of Therapeu tika is present and of course they are generally for an unlimited number suitable for indications. Without using the invention Wanting to limit particles to this is useful for special indications. Another object of the invention is there forth the use of the particles of the invention for producing a Medicines for cancer treatment, for the treatment of infectious diseases units and parasitoses, for the treatment of diseases and symptoms with a central nervous cause, for use in gene therapy or for genomic targeting. Another example is the use with the targe ting of cytostatics on tumor cells, in the transport of therapeutic usable substances through the blood-brain barrier and in the treatment treatment of serious infections (especially from eukaryotes) prefers.  

Other possible uses include e.g. B. the transfer of vegetable alkaloids with microbicidal activity in trypanosomes and of antioxidants and anti-inflammatory compounds [vitamin E, Bile acid, N-acetyl-L-cysteine, 2,6-bis (tert-butyl) -4-mercaptophenol, I buprofen and gentisic acid] in (degenerative) brain diseases, the Transfer of substances in hepatocytes, primarily for the treatment of Neoplasms, also increasing the effect of primaquine on the in the plasmodium hypnozoites that persist in liver cells. Are effective the particles according to the invention also against Trypanosoma brucei brucei.

Among the possible applications for the particles according to the invention, the following should also be mentioned in particular:

• - Transport of otherwise non-brain-accessible pharmaceuticals (e.g. cytostatics, Psychotropic drugs, pain relievers, M 'Alzheimer's therapeutics) with anti body-conjugated carriers through the blood-brain barrier.
• - Targeted introduction of pharmaceuticals (e.g. antivirals, cytostatics, Plasmodicides) with glycoside-conjugated carriers in hepatocytes.
• - Oral administration of otherwise only parenterally available pharmaceuticals by targeting intestinal epithelia.
• - Increase the effect of antiparasitic therapeutic agents through taring ting on parasite-specific surface molecules.

Another object of the method is the treatment of a Human or animal who needs this treatment with or using the particles according to the invention. Is particularly suitable this treatment for the above indications and application ten.

In the following section, the invention is further illustrated by examples tert without restricting them to it.

Examples and illustrations Illustrations

Fig. 1 shows schematically the general structure and shape of particles according to the invention, simple particles, simple stealth particles, target-seeking actinospheres and target-seeking acanthosphere.

Fig. 2 shows the synthesis of ktenates.

Fig. 3 shows the overall structure of a finished monomolecular particle with spikes, also called "Bdellosom" (Greek bdella = leech).

Fig. 4 shows an electron micrograph of BSA-conjugated ktenate particles (acanthospheres).

Fig. 5 shows the results of a model test on the unicellular parasite Trypanosoma brucei. The binding of particles produced according to the invention to the target cells correlates with the parasiticidal action of the contained daunomycin.

Examples General remarks

The examples set out below describe particles encompassed by the invention, in particular nanoparticles, in which it is possible to incorporate active substances into colloidal carrier particles. These can optionally be linked, for example, with antibodies against or natural ligands for characteristic molecular structures of the target or other “searcher molecules”. Optionally, the nanoparticles can be protected against the immune system at the same time, for example, by inert coating of their surface. In general, the particles encompassed by the invention described here by way of example are colloidal, lipid-free particle systems. Some of the particles described here and included in the invention are referred to below with the general terms actinospheres and acanthospheres (see Fig. 1). Because they denote structural concepts and non-steric basic shapes, these terms are retained regardless of the non-steric basic shapes, regardless of the actual geometry.

Because of their shape, the particles are generally referred to as Bdellosomes draws.

example 1 Production of the Bdellosome body a) General description of the basic body

The example is based on monomolecular particles from a complex structured polylactide derivative, due to its comb structure as ctenate designated. In an aqueous environment, ktenate forms filiform particles, so-called called Bdellosomen, with free choice by varying the synthesis parameters dimensions (diameter in the nanometer range, length up to several micrometers), hereinafter called Bdellosome (gr. Bdella = Leech). It is capable, on the one hand, of low molecular weight, preferably hyd rophobic substances to be stored stably, on the other hand in exposed functions to be chemically modified in such a way that "targeting" can be reached. The particle systems are implemented by Synthesis of monomolecular particles based on polyvinylal alcohol as the "backbone", on the OH groups of which chains made of polymer Lactic acid (or another suitable hydroxycarboxylic acid) be densified. The length of the polyvinyl backbone therefore determines the  Size of the particle in one dimension (longitudinal axis) and will follow referred to as a.

Since the side chains consist of bifunctional monomers - each of these monomers has an OH end to which the next acid molecule (either a similar monomer or a terminating molecule of non-hydroxycarboxylic acid can attach) and a COOH end that ends with a another hydroxyl group (either that of a monomer of the same type or an OH group of the polyvinyl backbone which leads to the end of chain growth), it is possible to provide “end pieces” by adding small but mutually equimolar amounts of acid-free alcohol and non-hydroxylated carboxylic acid whose concentration regulates the chain length relative to the concentration of the hydroxycarboxylic acid. The alcohol groups of the polyvinyl backbone serve as the COOH-side end piece and any other carboxylic acid as the OH-side end piece. Of course, the molarity of the carboxylic acid to be added at the OH-terminal must correspond to the molarity of the OH groups of the polyvinyl alcohol in order to create defined reaction conditions. This results in the following ratio b, which corresponds to the average chain length of the side chains and, together with a, defines the geometry of the nanoparticles that are formed:

b = c (hydroxycarboxylic acid): {c (OH groups) = c (non-hydroxy Carboxylic acid)}

The synthesis is carried out by reacting the reaction mixture of polyvinyl alcohol, hydroxycarboxylic acid and non-hydroxy carboxylic acid in a water-free environment with thionyl chloride, which converts the acid groups into the corresponding chlorides, which then react with hydroxyl groups to form water and form polycondensates (see Fig . 2).

The result is a comb-shaped molecule with a side chains hanging from the polyvinyl backbone with an average length of b monomer units, which ends with a non-hydroxylated end piece. If suitable monomers are selected, this arrangement is energetically unfavorable in the aqueous environment and leads to the "rolling up" of the molecular structure to a spatial structure in which the side chains loop around the backbone on all sides ("bottle brush" structure). The backbone is largely stretched out by the side chains, so that the dimensions of the nanoparticle are in the following range:
Length: a * length of the polyvinyl monomer
Width and height:
√b. * Length of the side chain monomer.

A (e.g. with FMOC) protected A serves as the end piece of the side chains amino acid, the protective group can be removed at the end of the synthesis reaction can be split (in the case of FMOC by treatment with catalytic Amounts of piperidine), creating side chains with terminal, in aqueous  Milieu of spatially exposed amino group can be obtained, not only stabilize the structure through their hydrophilicity and flocculation in aq prevent medium, but also as starting points for O surface reactions (see Example 3) can serve.

b) Specific implementation

From suitable amounts of polyvinyl alcohol 200,000 (Mowi ol ™), lactic acid and N-FMOC alanine using pyridine as a solvent by reaction with thionyl chloride and subsequent Splitting off the FMOC group with piperidine creates molecules that act as Po lyvinyl (telo-alanyl polylactide) at [a = 4000, b = 100] nen. In the following, this class of substances with the general Be drawing acid-final-ktenate (a, b) (Greek kteés = comb) be be drawn.

The finished lactic acid alanine ktenate is washed with dichloromethane and then without further purification along with the substance of interest dissolved in an appropriate volume of benzyl alcohol. The solution is incubated overnight at room temperature, then saturated with water and finally taken up in a larger volume of water. Without further mechanical treatment (ultrasound) removes the droplets the organic phase within a few hours and form a homogeneous suspension of substance-laden ktenate particles.  

Due to the elongated shape of the ktenate particles, statements are Of course, it is difficult to find out about the dimensions of the particle population However, the QELS method showed that <95% the mass of a typical preparation (unfiltered) in the form of particles in the submicron range.

Example 2 Production of the actinospheres and acanthospheres a) General comments

The wide range exists in both actinospheres and acanthospheres Ren components from a bifunctional linker, especially one Polyethylene glycol molecule that has an amino-reactive group at one end at the other end a different group with a different reak activity. This bifunctional polyethylene glycol on the one hand Particles from steric blocking of the surface with an inert mole removed from the access of the immune system (similar was connection with liposomes under the name "stealth technique" tested) and further stabilized, on the other hand the possibility required, additional molecules that mediate the actual target specificity in spatially favorable and flexible position to attach to the particles. At these molecules can be micromolecules, e.g. B. sugar, in which case an exclusive use of the bifunctional poly ethylene glycol is possible (actinospheres), or to macromolecules like  about antibodies (acanthospheres), in which case it is steric It is advisable to establish only a small part of the functional groups implement the particle surface with bifunctional polyethylene glycol and the rest by monofunctional polyethylene glycol, which is then al the physical and immunological stabilization of the particles serves to saturate.

b) Specific implementation following example 1

To suppress immune reactions, the finished Bdelloso men covalently according to Example 1 via the functional surface groups linked with "spikes" made of polyethylene glycol (MW ~ 3400 Da), de distal ends again antibodies or others for targeting usable molecules can be added.

Conjugation of the particles with "spikes" from NHS-ester-PEG takes place through simple mixing and incubation at room temperature, preferably in weakly basic medium, followed by dialysis against 500-fold Vo lumen of water (with a MWCO of 12-14 kDa both bound PEG spikes as well as unconjugated ktenate particles, not ever but ktenate-PEG conjugates escape) for cleaning.

When using bifunctional NHSester PEG vinyl sulfone spikes includes the secondary conjugation of the ktenate-PEG complex the "viewfinder" proteins, followed by saturation of the possibly still free coupling groups with cysteine (approx. 1 mg cysteine per mg PEG accordingly  a 20-30 fold molar excess) and a second Purification step by renewed dialysis, this time using a dialysis membrane with a correspondingly higher MWCO.

All in all, it is recommendable, still unsaturated, free Kopp Saturation groups with cysteine or another SH reagent. It is therefore recommended that, after implementing the Bdellosomen PEG Conjugates with the "searcher" molecules may still have free coupling groups at the distal ends of the PEG spines with a high mola saturate ren excess of a suitable reactant, in the case of vinyl sulfone e.g. B. Cysteine. In this way it is prevented that in Organism these coupling groups with the body's own molecules (e.g. Serum proteins) react and thus the target specificity is falsified.

When using the slow-reacting, relatively water-stable vinyl sulfone group as the distal reaction group of the PEG spines it is possible these thiophilic (ktenate) particles for all applications in one standard procedures and only after purification to connect with the appropriate "finder molecules", any of which can be chemical in nature and have only one sulfhydryl group zen so that actinospheres and acanthospheres are produced can. In acanthospheres, a simple in-process control of the last th linking step by adding fluorescently labeled or self fluorescent proteins (GFP) or by Western blot of  Samples possible, for actinospheres by detection in the thin layer chromatography.

Example 3 Properties of Bdellosomes prepared according to Example 1

The stability and packaging efficiency of Bdellosome are extremely high compared to other colloidal packaging systems. The loading efficiency was examined using the example of the clinically used cytostatic daunomycin. With the model substance Daunomycin, packaging rates of over 80% of the 06375 00070 552 001000280000000200012000285910626400040 0002010118852 00004 06256 material were achieved. When using a small amount of ³ H-labeled Dau nomycin, a yield of up to 99% of the total amount used was achieved.

Even after storage for several months at room temperature, the particles showed no signs of disintegration or flocculation. The simplicity of the manufacture of the Bdellosome, which results from the monomolecular particle structure, should also be emphasized. The shape and size of the particles could be made visible in the scanning electron microscope after weak gold vapor deposition and met the expectations (see Fig. 4); When using BSA, which has several free thiol groups, the particles were expected to crosslink slightly. With the functional surface groups covalently linked with "spikes" made of polyethylene glycol (MW 3400), at the distal ends of which antibodies or other molecules that can be used for targeting can be added, a reduction in the undesired uptake of the particles by the reticuloendothelial system was achieved shown on the rat model.

Example 4 Effectiveness of Bdellosomes Prepared According to Example 2 / Control of the parasitic unicellular organism Trypanosoma brucei brucei through acanthospheres loaded with daunomycin a) For the selection of the model organism

Trypanosoma brucei brucei is a Protozoan (Ord. Kinetoplastida) Pa rasit, who himself is not pathogenic to humans, but both because of him induced Nagana disease of African livestock is of direct economic importance as well as a laboratory model for the control of closely related human pathogenic forms Trypanosoma cruzi (Chagas disease, <20 million infected in South America), Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense (sleeping sickness, large prevalence in Central African Population) and some smaller trypanosome species (T. equinum, T. equiperdum, T. evansi) and the closely related Leishmania (Leish mania donovani, causative agent of the Kala Azar or visceral melaniasis; L. tropica, cause of the oriental bump; L. brasiliensis, mucous membrane Leishmaniasis) can be used.  

The treatment of parasitic protozoa is generally difficult and based mainly on suramin, pentamidine and organic arsenic and anti mon compounds such as melarsoprol and stibophen, their tolerability is bad at the concentrations required for therapy. Furthermore ge in the late stage of infection, the trypanosomes pass into the CNS, where they through the blood-brain barrier of the host of chemotherapy largely be withdrawn. Trypanosomes are therefore doubly suitable as models, on the one hand for direct targeting, on the other hand for transfer via the blood Brain barrier. Direct targeting is described below.

Trypanosomes elude their host's immune system by right combinative variation of their cell surface proteins, only a few of which Receptors (inter alia for transferrin and albumin) are excluded, which however, are positioned in wells of the cell surface such that binding antibodies no Im leading to the destruction of the parasite cell can initiate munreaction. However, it is possible in principle with cell poison loaded bilosomes with ligands for these receptors and endocytose in this way.

b) Production of the acanthospheres based on the Bdellosome

Bdello som suspensions (0.5 g lactic alanine ktenate 4000, 100 per liter) with a loading of 1% (m / m) daunomycin (corresponding to a concentration of 10 ^-5 mol daunomycin per.) Were used for both binding and cytotoxicity studies 1 suspension), which had a tritium label of approx. 1000 cpm / µg. Free daunomycin from a concentration of 10 ^-7 mol in the medium hinders the growth of trypanosomes.

The particles were prepared as described in Example 1. The Bdellosomes were prepared according to Example 1 via their surface Amino groups either linked to monofunctional PEG or via bifunctional (NHS ester / vinyl sulfone) PEG with a medium mole molecular weight of 3400 Da with cysteine residues of various proteins couples: human transferrin in various concentrations, Rinderse rumalbumin and single-chain antibodies against the transferrin receptor. Finally, free coupling groups were saturated with cysteine.

c) attachment studies

In each case ~ 3.10 ⁵ parasites in good growth were incubated for 20 min at room temperature with a 1: 1 mixture of one of the above-described Bdellosomen suspensions and growth medium, then the supernatant was drawn off, the cells were washed in isotonic saline solution and finally in fresher Saline solution added. The tritium activities in the supernatant, wash buffer and cellular fraction were determined by scintillation counting.

d) cytotoxicity studies

In each case 10 μl of the above-described Bdellosomen suspensions were added to a suspension of ~ 10 ⁵ parasites in good growth in fresh nutrient medium (so that a final concentration of 10 ^-7 mol was reached) and incubated under growth conditions overnight, counted after 24 hours, and the cell numbers of the parasites were compared with those of the controls treated with isotonic saline only.

e) conclusion

The results of the binding and cytotoxicity studies show a cor relation between cytotoxicity and binding of <97%. The strongest binding fraction (antibody-related Bdellosome) decreased the cell density of the parasites to a quarter of that only with isotonic sodium chloride solution treated control, d. that is, it became a pronounced cytotoxi observed effect. In the absence of "seeker" proteins, however were neither binding nor cyto at the same daunomycin concentration observing toxicity.

Claims (36)

1. Containing monomolecular solid particles for the transport of hydrophobic or hydrophobized active substances

• a) an unbranched or a maximum of three branched moles of the cooling backbone made of a polymer composed of monomers with at least one binding group (x) on each monomer, the binding groups (x) being covalently linked via a (x) - (x ') bond
• b) polycondensed molecular side chains composed of chain-forming monomers each having at least one binding group (y) and at least one binding group (x ') or composed of different chain-forming monomers, of which one monomer has at least two binding groups (y) and another monomer has at least two bonds groups (x '), or built up from various chain-forming monomers, one monomer of which has at least two binding groups (y), another monomer of which has at least two binding groups (x') and a further at least one binding group (y) and at least one binding group (x '), where (x') can form a covalent bond with (x) and also a covalent bond with (y), where at the end of the molecular side chains in each case covalently via a (y) - (y ') bond
• c) side chain end pieces which have at least one binding group (y '), no binding group (y) and at least one free group (z) optionally provided with a protective group, are bound,

in which
the molar ratio between the monomers of the molecular side chains (b) and the side chain end pieces (c) is << 1,
is the group y ≠ of the group z and the group x '≠ of the group z,
the molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
x, x ', y and y' are independently selected from OH, SH, COOH or N / H ₂ under the condition that x / x ', x' / y and y / y 'respectively correspond to bond pairs x / x' , x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -OC (O) -), NH ₂ / COOH (amide bond - NH-C (O) - ), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH-C (O) - ) or COOH / SH (thio-ester bond -SC (O) -) and
z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group. 2. Particles according to claim 1, characterized in that the parti non-covalently bound, hydrophobic or hydrophobized contain active pharmaceutical ingredient. 3. Monomolecular solid particles for transporting hydrophobic or hydrophobized active ingredients can be prepared by a process in which unbranched or a maximum of three times branched polymer backbone composed of monomers with at least one binding group (x) on each monomer

• a) chain-forming side-chain monomers each having at least one binding group (y) and at least one binding group (x '), where (x') can form both a covalent bond with (x) and with (y),
• b) a mixture of chain-forming side chain monomers, of which at least one monomer has at least two binding groups (y) and at least one other monomer has at least two binding groups (x '), where (x') is both a covalent bond with (x) and can enter with (y), or
• c) a mixture of chain-forming side chain monomers, of which at least one monomer has at least two binding groups (y), at least one other monomer has at least two binding groups (x ') and at least one further monomer has at least one binding group (y) and at least one binding group (x' ), where (x ') can form a covalent bond with (x) as well as with (y),

and at least one side chain end piece, with at least one binding group (y '), without binding group (y) and with at least one free group (z), optionally provided with a protective group, where (y') can form a covalent bond with (y) ,
is brought into contact under conditions which allow a bond between the monomers or monomer mixtures of the side chains and the polymer backbone and the chain end pieces and also a polycondensation of the monomers or monomer mixtures of the side chains,
in which
the molar ratio between the monomers of the molecular side chains (b) and the side chain end pieces (c) is << 1,
is the group y ≠ of the group z and the group x '≠ of the group z,
the molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y and y / y 'respectively correspond to pairs of bonds x / x', x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -O- C (O) -), NH ₂ / COOH (amide bond -NH-C (O) -), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH-C (O) -) or COOH / SH (thioester bond -SC (O) -) form,
z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the Vi nyl or epoxy group, optionally protected by a protective group, and
the side chain monomers can be used as pure monomers or derivatives such as intramolecular anhydrides or lactones, as long as they can still form chains with themselves and / or other side chain monomers. 4. Monomolecular solid particles for the transport of hydrophobic or hydrophobized active substances can be prepared by a process in which unbranched or a maximum of three branched polymer backbone composed of monomers with at least one binding group (x) on each monomer
polycondensed molecular side chains composed of monomers each having at least one binding group (y) and at least one binding group (x ') or composed of different monomers, of which one monomer has at least two binding groups (y) and another monomer has at least two binding groups (x'), or built up from different monomers, where one monomer has at least two binding groups (y), another monomer has at least two binding groups (x ') and another has at least one binding group (y) and at least one binding group (x'), where ( x ') can form a covalent bond with (x) as well as with (y), where at the end of the molecular side chains in each case covalently via a (y) - (y') bond, side chain end pieces which have at least one binding group (y '), have no binding group (y) and at least one free group (z) optionally provided with a protective group, where (y ') can form a covalent bond with (y) ,
is brought into contact under conditions which allow a bond between the polycondensed molecular side chains and the polymer,
in which
the molar ratio between the monomers of the molecular side chains (b) and the side chain end pieces (c) is << 1,
is the group y ≠ of the group z and the group x '≠ of the group z,
the molar ratio between the monomers of the molecular backbone and the side chain end pieces (c) is approximately equimolar,
x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y and y / y 'respectively correspond to pairs of bonds x / x', x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -O- C (O) -), NH ₂ / COOH (amide bond -NH-C (O) -), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH-C (O) -) or COOH / SH (thioester bond -SC (O) -) form and
z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the Vi nyl or epoxy group optionally protected by a protective group. 5. Solid particles for transport according to one of claims 3 or 4, characterized in that the contacting in one anhydrous organic solvents, preferably pyridine takes place.   6. Solid particles for transport according to one of claims 3 to 5, characterized in that the contacting in counter was from thionyl chloride. 7. Particles according to one of claims 3 to 6, characterized records that protective groups are split off if necessary. 8. Particles according to one of claims 3 to 7, characterized records that there is a washing step, preferably with dichlor methane, connects. 9. Particles according to one of claims 3 to 8, characterized records that the product then together with the transporting hydrophobic or hydrophobized pharma active ingredient in an anhydrous organic solvent solvent is dissolved, then the solution for some time, preferably it is incubated overnight, preferably at room temperature, then the solution is saturated with water and then the water-saturated solution in a larger volume of water is resolved, mechanical treatment if necessary closes and then optionally cleaned the particles and iso be lated. 10. Particles according to claim 9, characterized in that after the solution of the water-saturated solution in a larger volume of water there is no mechanical treatment and / or the cleaning by dialysis preferably takes place against H ₂ O. 11. Particles according to one of claims 9 or 10, characterized in that the water-free organic solvent dissolves in water in the ratio solvent: water between 1:10 and 1:50, preferably between 1:20 and 1:40, in particular between 1: 20 and 1:30 solves, and / or is preferably selected from:
Methylene chloride or benzyl alcohol, preferably benzyl alcohol. 12. Particles according to one of claims 1 to 11, characterized indicates that the polymer backbone is unbranched or maximal times branched, preferably unbranched. 13. Particle according to one of claims 1 to 12, characterized in that
the monomers of the side chain each have a maximum of two groups (y) and a maximum of two groups (x ') and / or
the group (y) in the monomers of the side chain corresponds to the group (x) in the polymer backbone and / or
the group (x ') in the monomers of the side chain corresponds to the group (y') in the side chain end piece and / or
the group (z) is selected from the "free groups" OH, SH, COOH or NH ₂ and the vinyl or epoxy group, optionally protected by a protective group,
the monomers of the side chain each have a maximum of 2 to 10 carbon atoms, preferably 2 to 6 carbon atoms, in particular 2 to 4 carbon atoms, and / or
the monomers of the side chain which have both the group (y) and the group (x '), either only 1 group (y) and 1-2, preferably 1, groups (x') or only 1 group (x ') and 1-2, preferably 1, groups (y) and / or
the monomers of the side chains are identical except for the side chain end piece with 1 group (y) and 1 group (x ') or the monomers of the side chains are constructed identically and alternately monotonously up to the side chain end piece from alternately a monomer with 2 groups (x' ) and a monomer with 2 groups (y). 14. Particle according to one of claims 1 to 13, characterized in that the polymer backbone is selected from
Polyvinyl alcohol, polyacrylic acid, polyvinylamine, polysaccharide or polyamino acid,
preferably polyvinyl alcohol or polyacrylic acid,
especially polyvinyl alcohol. 15. Particle according to one of claims 1 to 14, characterized in that the monomers of the side chain are selected from
Hydroxycarboxylic acids, amino acids, the combination of diamines and dicarboxylic acids or the combination of diols and dicarboxylic acids, or their derivatives,
preferably hydroxycarboxylic acids or the combination of diols and dicarboxylic acids or their derivatives,
in particular hydroxycarboxylic acids such as lactic acid, glycolic acid, tartaric acid or citric acid or their derivatives. 16. Particle according to one of claims 1 to 15, characterized in that the side chain end pieces are selected from
unprotected amino acids, N-protected amino acids, COOH-protected amino acids, unprotected amino alcohols, N-protected amino alcohols, O-protected amino alcohols, unprotected thiol alcohols, O-protected thio alcohols, S-protected thio alcohols or unprotected thiol acids, S-protected thiol acids, CO protected thiol acids, unprotected thioamines, S-protected thioamines or N-protected thioamines,
preferably
unprotected amino acids, N-protected amino acids, unprotected amino alcohols, N-protected amino alcohols, unprotected thiol alcohols, S-protected thio alcohols or unprotected thiol acids, S-protected thiol acids,
in particular
unprotected amino acids, such as alanine, N-protected amino acids, such as N-FMOC-β-alanine, unprotected thiol acids or S-protected thiol acids. 17. Particle according to one of claims 1 to 16, characterized in that the polymer backbone, the monomers of the side chain or their derivatives and the side chain end piece or its derivatives are selected from one of the following combinations:
preferably
in particular
18. Particle according to one of claims 1 to 17, characterized shows that the particles have a length of <5 µm, preferably <3 µm, in particular <2 µm and / or a thickness and width of < 200 nm, preferably <75 nm, in particular <30 nm. 19. Particle according to one of claims 1 to 18, characterized in that linker molecules which have a reactive group (z ') selected from groups which are selected with one of the groups (z) from the "free groups" (z ) selected from OH, SH, COOH or the NH ₂ and the vinyl or epoxy group can enter into a covalent bond, preferably an amino- or thiol-reactive group, in particular an amino-reactive group, have, covalently via (z ') - (z) bonds with groups (z) present on the surface of the particle selected from the “free groups” to which particles are bound. 20. Particle according to claim 19, characterized in that the linker molecules are bifunctional and, in addition to the reactive group (z ') binding to the particle according to one of claims 1 to 18, also a further reactive group at another end of the molecule ( z "), selected from reactive groups, with one of the groups (z) selected from the" free groups "(z) selected from OH, SH, COOH or NH ₂ and the vinyl or epoxy group enter into a covalent bond may have, preferably a thiol-reactive group, where z '≠ z ". 21. Particle according to one of claims 19 or 20, characterized records that the linker molecules are a mixture of bifunctional Molecules according to claim 20 and monofunctional molecules, which in addition to the particle according to one of claims 1 to 18 binding reactive group (z '), preferably the amino or thiol-reactive, especially the amino-reactive, group on kei at the other end of the molecule another, differently reactive have functional group (z ") with z '≠ z" are. 22. Particles according to claim 21, characterized in that at the Surface of the particles significantly more, preferably at least 100% more, monofunctional than bifunctional linker Molecules are covalently bound. 23. Particle according to one of claims 20 to 22, characterized records that bioactive to the bifunctional linker molecules Macromolecules or "finder" molecules selected from Pepti the, proteins; preferably antibodies, antibody fragments or antibody derivatives with target binding properties such as "Single-chain" antibodies; Hormones, sugars, preferably Glycosides; synthetic or natural receptor ligands; Proteins or peptides with a free cysteine group or thi ozucker, coupled via a bond to the reactive group (z ") pelt, are coupled or before the surface modification tion were coupled.   24. Particles according to one of claims 20 to 22, characterized records that bioactive to the bifunctional linker molecules Macromolecules or "finder" molecules, preferably antibodies per, antibody fragments or antibody derivatives with target binding the properties like "single chain" antibody, in particular with free cysteine group, via a bond to the reactive group (z ") are coupled, are coupled or in front of the surface Chen modification were coupled. 25. Particles according to claim 20, characterized in that to the bifunctional molecules of the coating bioactive micromo leküle or "Sucher" molecules, preferably sugar, in particular their thio sugar; Peptides or hormones, especially those with free ones Cysteine group, via a bond to the reactive group (z ") are coupled, coupled or in front of the surface mo dification were coupled. 26. Particles according to one of claims 23 to 25, characterized characterized in that after binding the bioactive micromolecules or "searcher" molecules still free reactive groups (z ") be saturated, preferably with cysteine. 27. Particles according to claim 19, characterized in that the Linker molecules are monofunctional molecules that are next to the binding to the particle according to any one of claims 1 to 18 reactive group (z '), preferably the amino or thiol reactive  Group, especially the amino-reactive group, in none another end of the molecule, another, differently reactive group (z ") with z '≠ z". 28. Particles according to one of claims 19 to 27, characterized records that the linker molecules are polyglycolides, preferably polyethylene glycol derivatives, especially NHS Ester polyethylene glycol or NHS ester / vinyl sulfone Polyethylene glycol. 29. Particles according to one of claims 1 to 28, characterized records that the active ingredient to be transported is a synthetic or natural active ingredient, a protein, peptide, lipid, sugar or Nucleic acid or a low molecular weight organic or high molecular organic active ingredient, for example a hormone, an antineoplastic substance, an antibiotic, antifungal cum, parasiticide, antiviral or antihelmintic, a cardi ovular active substance; a central active substance, esp especially an analgesic, antidepressant or antiepileptic; is. 30. Particle according to one of claims 1 to 18, characterized in that it is directly or via a linker; preferably via bifunctional polyethylene glycol molecules; is linked to a "viewfinder" molecule selected from:
Peptides, proteins; preferably antibodies, antibody fragments or antibody derivatives with target-binding properties such as "single-chain"antibodies; Hormones, sugars, preferably glycosides; synthetic or natural receptor ligands; Proteins or peptides with a free cysteine group or thio sugars. 31. A method for producing a particle according to one of claims 1 or 2, characterized in that an unbranched or a maximum of three branched polymer backbone (a) composed of monomers each having at least one binding group (x) on each monomer
Side chain monomers (b) each having at least one binding group (y) and at least one binding group (x '), where (x') can form a covalent bond with both (x) and (y),
a mixture of side chain monomers (b), of which at least one monomer has at least two binding groups (y) and at least one other monomer has at least two binding groups (x '), where (x') is a covalent bond both with (x) and with (y) can enter, or
a mixture of side chain monomers (b), of which at least one monomer has at least two binding groups (y), at least one other monomer has at least two binding groups (x ') and at least one further monomer has at least one binding group (y) and at least one binding group (x '), where (x') can form a covalent bond with both (x) and (y),
and at least one side chain end piece (c), with at least one binding group (y '), without a binding group (y) and with at least one free group (z) which may be provided with a protective group,
in which
the molar ratio between the monomers of the molecular side chains (b) and the side chain end pieces (c) is << 1,
is the group y ≠ of the group z and the group x '≠ of the group z,
the molar ratio between the monomers of the molecular backbone (a) and the side chain end pieces (c) is approximately equimolar,
x, x ', y and y' are independently selected from OH, SH, COOH or NH ₂ under the condition that x / x ', x' / y and y / y 'respectively correspond to pairs of bonds x / x', x '/ y or y / y' (with the corresponding bond) selected from OH / COOH (ester bond -O- C (O) -), NH ₂ / COOH (amide bond -NH-C (O) -), SH / COOH (thio-ester bond -SC (O) -), COOH / OH (ester bond -OC (O) -), COOH / NH ₂ (amide bond -NH-C (O) -) or COOH / SH (thioester bond -SC (O) -) form,
z is selected from CH ₃ , OH, SH, COOH or NH ₂ and the Vi nyl or epoxy group optionally protected by a protective group and
the side chain monomers (b) can be used as pure monomers or derivatives such as intramolecular anhydrides or lactones, as long as they can still form chains with themselves and / or other side chain monomers,
is brought into contact under conditions which allow a bond between the monomers or monomer mixtures of the side chain and the polymer backbone and the side chain end pieces and also a polycondensation of the monomers or monomer mixtures of the side chain,
optionally in an anhydrous organic solvent, preferably pyridine, optionally in the presence of thionyl chloride and then optionally purified, preferably by dialysis against H ₂ O, and optionally isolated
and then optionally the particles with the hydrophobic or hydrophobized active ingredient to be transported are dissolved in an anhydrous organic solvent, then the solution is incubated for some time, preferably overnight, preferably at room temperature, then the solution is saturated with water and then the water-saturated solution is dissolved in a larger volume of water, optionally followed by a mechanical treatment and then optionally cleaned the particles, preferably by dialysis against H ₂ O, and isolated who the. 32. A method for producing a particle according to any one of claims 19 to 29, characterized in that a particle according to one of claims 1 to 18 containing a group (z) selected from the free groups with a linker molecule containing a reactive group (z '), which can form a covalent bond with the group (z), brings them into contact under conditions suitable for the formation of this covalent bond and then optionally cleans the particles, preferably by dialysis against H ₂ O, and isolates them. 33. A method for producing a particle according to any one of claims 23 to 25, characterized in that particles produced thereafter, which have a free reactive group (z ") with bioactive on bifunctional linker molecules following the method according to claim 32 25 bringing macromolecules or "finder" molecules according to one of claims 23 to 25 into contact under suitable conditions to form a bond between the group (z ") and the bioactive macromolecules or" finder "molecules and, if appropriate, subsequently cleaning the particles, preferably by dialysis against H ₂ O, and isolated. 34. Medicament containing particles according to one of claims 1 to 30 and any suitable additive and / or auxiliary fe. 35. Diagnostic agent containing particles according to one of claims 1 to 30 and any suitable additive and / or auxiliary fe. 36. Use of particles according to one of claims 1 to 30 for the manufacture of a medicament for cancer treatment, for loading treatment of infectious diseases and parsitosis, for treatment treatment of diseases and symptoms with central nervous ursa che, for use in gene therapy or for genomics Targeting.