GB2507983A - Mesoporous silica particles and their use in drug delivery - Google Patents

Abstract

A substantially monodisperse assemblage of particles 10 having interconnected nanopores 20 and a core 30 with a shell 40 disposed about the core 30 wherein the particles 10 have a cubic crystal form. In another aspect, a method for the sustained release of an active substance to an environment comprising adding an active substance to the particles 10, placing restrictions 50 in the interconnected nanopores 20 within the shell 40 by covalent bonding (preferably using dextran polymers) and placing the particles with the added active substance in the environment. The particles are preferably either nanoprous or mesoporous silica. Alternatively, a method for the manufacture of a plurality of monodisperse particles 10 comprising: mixing an ionic surfactant (preferably cetyltriethylammonium bromide) with an inorganic salt (preferably sodium sulfate) in hydrochloric acid; mixing a co-solvent with a nonionic surfactant (preferably a polyalkylene oxide) and an oxide source (preferably TEOS); and mixing the two together.

Description

Title Susta ed-r&ease Formubtion R&d of the Invention [0001] The present invention discloses a material comprising an assemblage of particles releasing active substances, such as? but not limited to, pharmaceuticals, over a prolonged period of time foliowing a zero order kinetic.

ndofthejnventjon [0002] The sustained-release of active substances (AS) over long tim&scales is a desirable characteristic in many areas such as crop science, medicine, cosmeflcs, etc. The term "active substance" used in this context denotes any substance that fulfils a specified function. The active substance can be, for example, a biocide, a pharmaceutical, a perfume or flavour, a fertUizer, or a plant hormone. For sustained release, the active substance is dispensed or distributed in a supporUng material, so that it slowly dissolves or diffuses into the specified environment. Examples of such formulations are known, for example, from US Patent Application Publication No US 2009/0165515A1, European Patent Application No EP021814$A1 and US Patent No. 3994439.

[0003] Sustainedrelease formulations in medical applications can control rate and period of a drug delivery to a certain degree. It is known that traditional therapies with repeated drug administration result in a saw-tooth curve of drug concentration in the bloodstream. The sustainedrelease formulations enable keeping the drug concentration in a so-called "therapeutic window" for a prolonged time span. Polymers, such as PLGA (poiy-(lactic-co.-glycolic acid)) are common carriers tar such sustainedrelease formulations, as described in "Polymeric Delivery Systems for Cbntro/led Drug Re/ease", RLanger, Chem, Eng. Commun. 6 (1980) 1-3, 1. Several modifications of the sustained-release formulation were introduced in order to expand the timescale of the therapeutic window to several weeks or months by reducing diffusion of the active substances. Successful examples of such sustained-release formulations are intercalation of Inert nanoparticles [as known from US Patent No 6,821,928] or microencapsulation of the AS [as know from US Patent No 6,265,3893.

[0004] A typical release pattern for slow-release systems Is first-order kinetics, In which release rate of the active substance decreases exponentially with time, with relatively high Initial release rate. High initial concentrations of some drugs in the bloodstream can cause toxic side effects. After a certain period of time, the concentration of the drug In the bloodstream falls below the necessary therapeutic level [see for a discussion "Nanostructure-medlated drug deI1very G. A. Hughes, Nanomedicine: Nanotechnology, Biology, and Medicine 1 (2005) 22]. In contrast, with a zero-order kinetic release pattern for the active substance a substantially steady therapeutic level can be maintained over the treatment period. This is preferably done with only a single administration of the active substance.

[0005] The terms "zero-order release" and "zero-order kinetiC are to be understood in this context as a release pattern of the active substance from a substrate over time, In which the first temporal derivation of the release rate is substantially zero, or, In other words, the release rate remains substantially constant with time. Similarly, the term "first-order release" or "first-order kinetiC Is to be understood as a release pattern over time, in which the first temporal derivation of the release rate has a substantially fixed, time-independent value.

(0006] In the field of sustained-release formulation, nanoporous materials have drawn much attention as the nanoporous materials are suitable as supporting "host" materials for specific active substances. The ordered nanoporous materials are mainly based on silicon oxide, and, to a lesser extent, on other metal oxides, and comprise a specific oxide with a regular arrangement of pores [see, for example, "Ordered mesoporous materials" U. Ciesla and F. Schueth, Microporous and Mesoporous Materials 27(1999) 2-3, 131-149].

[0007] The term "nanoporous material" (or oxide, silica, etc.) used in this disclosure Is to be understood as a porous material with pore diameters substantially between 1 and 100 nm. .3-

10008] The term "mesoporous material" used in this disclosure Is to be understood as a nanoporous material with pore diameters substantially between 2 and 50 nm (see J. Rouqueroi et al., Recornmendatlons for the characterization of porous solids (Technical Report)", Pure & Appl. them 66 (1994)8 1739-1758. dol:10.1351/pac199466081739).

[00091 The term "monodisperse" as used in this disclosure refers to a collection of particles that are substantially of the same size, shape and mass.

(00010) It Is known that porous silica (Si02) is a non-toxic, biocompatible material that can incorporate a high volume of active substances into its open pore system. [regarding the biocompatibility see, for example: "Unique Uptake of Acid-Prepared Mesoporous Spheres by Lung Epithelial and Mesothellon,a Cells S. Blumen et aL, American Journal of Respiratory and Molecular Biology vol. 36 (2007), pp. 333-342]. A further advantage of this class of materials, and more particularly, mesoporous ordered silica, is its extreme versatility regarding the shapes and sizes of Its pore systems. The pore system can be controlled during the synthesis, thus making various pore sizes and geometries available.

(00011] Various structures of the silica materials with different pore geometries are commonly classified by a three-letter code followed by a number. A list of available structures can be found, e.g. in US Patent 7,767,004 62, Table 1. Additionally, various functional organic groups can be selectively introduced onto the outer and Inner surfaces [see "Mesoporous Materials for 0mg DelIveriJt M. Vallet-Regi et al., Angew. Chem. mt Ed.

46(2007) 7548].

(00012] The sustained release formulations comprise at least two components, namely, the supporting or host material (sometimes called subildte), and the particular active substance. Different superstructures of the two components are therefore imaginable. One superstructure for the sustained release formulation with a zero order kinetic is a "coated pure drug bead", which has a bead exhibiting a core-shell structure. The core is formed by the pure active substance, and the shell Is formed by a second, supporting material, (00013] The theoretical release behaviour of such core-shell structures is described In "Dimensionless presentation for drug release from a coated pure drug beat S.M. Lu, Int, 1.

of Pharmaceutics 112 (1994), 105-116. It can be derived from this article that a zero-order kinetic sustained release from of a single bead can principally be achieved, if the following three preconditions are fulfilled; * The concentration of the active substance at the border of core to shell remains constant over a prolonged timespan.

* The diffusivity of the active substance in the core is much higher than its diffusivity In the shell.

* The concentration of the active substance In the surrounding medium of the particle remains zero or negligibly small (perfect sink).

(00014] All three preconditions might be, In principal, fulfilled by use of the coated pure drug beads, he. the core-shell structure. However, the encapsulation of the pure drug (as the active substance) has disadvantages concerning, for Instance, the mechanical stability of core-shell structures during processing. Therefore, a entirely non-collapsible, rigid porous network such as a nanoporous silicate as a supporting materIal is helpful or often even necessary as, for example, described in US Patent Application No. 2003/175347A1.

(00015] The reported results from the coated pure drug beads can be adopted to the more general case of an active substance incorporated into a rigid porous medium. In this latter case, the relevant parameters, for example, the concentration of active substance at the core-shell transition are only weakly altered if the core Is highly porous, comprising interconnected channels and an isotropic diffusion behaviour (cubic crystal system), and completely filled with the active substance. Typical examples demonstrate that diffusion of low-molecular substances in the porous systems, and therefore, their elution into the environment are relatively fast and mostly completed within minutes, or, sometimes, hours.

[see "inclusion of ibuprofen in mesoporous templated silica: drug loading and release propeity", C. Charnay et al., European Journal of Pharmaceutics and Biopharmaceutics vol. 57(2004)3, pp. 533-540].

[00016] in the case of poorly water-soluble active substances, to which belong a vast number of pharmaceuticay important substances [see US Patent 6,576,264 81], a three pre-conditions outlined above are fulfilled, If the porous coreshell particle is loaded wfth a poorly water-soluble active substance and brought into an open biological environment, the biological environment will act as a sink for released molecules of the active substance. A steady concentration of the active substance at. the core/shell frontier of the particle for a prolonged tUne can therefore be assumed, since the porous structure wiU be fflled with water from the biological environment, This water acts as a transport medium for the solubilised molecules of the active substance, and keeps the concentration of the active substance at the shell substantially constant. The shefl its&f must be designed in a way that the shell strongly hinders the diffusion of the active substance.

[00017] It would be advantageous to incorporate a large number of the supporting materials in a carrier material, for example, in a polymer extrudate, instead of the preparation of a single large porous particle. The use of the single large porous particle involves the danger of a huge and unwanted overdose of the active substance in case of breaking, and, therefore, of uncontrolled fast release from this single large porous particle.

In contrast, in case of the breakage of the polymer extrudate containing a large number of small particles, only a small fraction of the particles would be destroyed, and the amount of the active substance released would be much smalier, For medical applications, the use of such an assemblage of particles is therefore preferable.

[00018) In other applications, for example, in crop science, a wide-area application of the small particles as individual reservoirs for the active substances is additionally advantageous in order to achieve a substantially constant concentration of the active substance (eg.

hiocides) in time and space. This allows a reduction of the total amount of the active substance per area unit, since any unnecessary local overdose in area or time can be avoided. Other examples may comprise glues, coatings and lacquers, in which the particles releasing, for example, a biocide can be incorporated and prevent the particular composition from fouling. 6.

[000191 To ensure that such desirable release kinetics from a single particle can be transferred to an assemblage of particles, the sze distribution of the particles in the assemblage must be substantially monodisperse and show only a sma standard deviation.

[00020] This requirement is shown, for example, in "Modelling of druqrelease from pot>'-disperse micmencapsulcted spherical port/c/cC, C. Sirotti et aL, I Microencapsulation, 19 (2002) 5, 603614. Ut can be even more dearly visualized if for each particle of a batch a perfect zero order kinetic is assumed, Le, a constant release of the active substance over time unUl the reservoir (core) is emptied, followed by a sudden and abrupt stop.

[00021] The amount of the active substance incorporated into the particle is directly oroportional to the volume of the core of the particle, which is related to the cube of the particle radius. The amount of released active substance per unit time is related directly to the suriace area of the particle, which is the square of the particle's radius multiphed by 4m.

Thus, not only the standard deviation (SDV) of the size of the cor&depot, but also the SDV of the amount of the active substance r&eased per time is strongly affected by the WV of the particle's diameter. For example, the volumes of the smallest particles (2 micrometres in diameter) and the biggest particles (23 micrometres in diameter) in a mixture (that corresponds in this case to 11,1 % deviation from a mean particle size of 215 micrometres) differ almost by the factor of two. It is thus obvious, that a broad size distribution of the particles results in a huge, undesired distortion of the aimed zero-order kinetics, Hence, the particle size distribution has to be as sharp as possible, 00022] The correlation, visualizing quantitatively the effect of a different SDV for assemblies of the particles, can be derived as follows (if a large core and a negligible thin shell is assumed, so that + rhn, where rco, is the radius of the core and rshe is the radius of the shefi), [00023] The amount of incorporated substance is assumed to be directly proportional to the available volume of the depot, hence the mass of incorporated AS is Eq. 1 [00024) The amount of the active substance released per time is proportional to the surface of one particle = Eq. z [00025] Separation and integration of Eq. 2 leads to fi tht = c4r2 d Eq. 3 Eq. 4 [00026] Combination with Eq. 1 results in = Eq S [00027] The Eq. 5 shows the time at which the core depot of the particle is emptied. It is linearly r&ated to the radius of the particle and dependent on the diffusion rate of the active substance through the shell, which correlates with constant c3 and, therefore, c4.

[00028] Since the standard deviation and its influence on the release properties is the most interesting, constants c2, c4 and c5 in this example are defined to be equal to 1, [00029] The size distribution ci the particles is given by the Gaussian distribution P(r = e( (\2 Eq. 6 where r is the particle radius, t is the mean radius, u is the standard deviation, 100030] The amount of the active substance r&eased by a the partides per time is given by the sum, and, hence, by the integral of P(t), multipfled by the surface area of the particles. 2 1

2 - r t Tn ";rrc *c z' [00031] The denominator reflects the overall amount of the active substance, to normalize the curves obtained for different standard deviations and mean particle sizes. To calculate the release rate at a given time, the starting point of the integral of the nominator has first to be found from Eq. 5. The result is a value for a radius, corresponding to the sizes of the particles that no longer contain the active substance at a time t. Hence, the integration is done for all the particles that still contain active substance. The amount of the particles inside a given dr is multiplied by c.4 r2, which gives the release rate (Eq. 2).

[00032] Fig 16 shows four different curves relating to four different standard deviations. It can be dearly seen, that a relatively large SDV leads to a distorted release curve, in comparison to an almost rectangular curve for SDVs that are smaller than 10% of the mean particle size.

Prior Art

[000331 The use of nanoporous and mesoporous silica materials in sustained-release systems has been extensively discussed in literature. However, the known materials and formulations have been shown to have disadvantages limiting their use as a host for active substances in sustained-release applications.

[00034] The literature on attempts to use these nanoporous and mesoporous silica materials as the hosts for sustained-release applications can be dMded into two major groups. The first group comprises rather simple approaches in which a potentially surface-modified host is loaded with the active substance. An overview of this approach can be found In "Mesoporous Silica: An Alternative Diffusion Contm lIed Drug Delivery System, Topics in Multifunctlonal Blomaterlals and Device?; J. Andersson et al., Ashammakhi, N., Ed.; E-book, (2008).

1000351 Example are also disclosed in: * "Inclusion of Ibuprofen in rnesoporous templated silica: drug loading and release property', C. Charnay at S., European Journal of Pharmaceutics and Blopharmaceutics 57(2004), 3,533-540; * "Controlled Drug Delivery System Based on Ordered Mesoparous Silica Matrices of Captoprll as Anglotensin-Converting Enzyme Inhibitor Drug", R. Popovici et al., Journal of Pharmaceutical ScIences 100 (2011), 2, 704-713; -9..

* "TunIng drug uptake and release rates through different morphok*gies and pare diameters of can/InS mesoprorous silica? V. Cauda et al., Microporous and Mesoporous Materials 118(2009) 435-442; * "3D cubic mesoporous silica microsphere as a corner jbr poorly soluble drug carved/to!" V. Mu et al., Microporous and Mesoporous MaterIals 147(2012), 94101.

(000361 Even though these nanoporous and mesoporous silica materials loaded with the active substance exhibit a retarded release kinetic, leaching of the active substance is still too fast, as the leaching Is based solely on the diffusion retardation Inside the nanoporous host channels of Individual nanoporous and mesoporous silica particles. Most of the active substances are eluted typically from the nanoporous and mesoporous silica particles after several hours. This rapid elution Indicates that It is not possible to realize a zero-order kinetic in the range of days, and certainly not In terms of weeks or months in such a basic system.

[00037) The release time and the kinetics can be improved if moulded paddings with dimensions of several mm are prepared from the individual nanoporous and mesoporous silica particles, as, for example, disclosed In * "A New Property of MCM-41: Drug Delivery System" M. Vallet-Regi et al., Chemistry of Materials 13 (2001), 308-311; * "Influences of Material Characteristics on Ibuprofen Drug Loading and Release Profiles from Ordered Micro-and Mesoporous Silica Motrices", .1. Andersson et al, Chem. Mater 16(2004)4160-4167 * "Mesoporous SBA-15 HPLC evaluation for controlled gentarnicin drug dellverf, A. Doadrio et al., Journal of Controlled Release 97(2004), 125-132.

[00038] However, in that case, the main advantage of a particle assemblage of the nanoporous and mesoporous silica particles, namely, the possibIlity of achieving a homogenous release of the particular active substance in space by a distribution of the nanoporous and mesoporous silica particles in a particular environment is lost.

[00039) The second group of materials comprises core-shell materials, in which more efficient release retardation Is achieved by adding a shell that hinders the leaching of the -10-active substance out of the reservoir core, as described in the theoretical section above.

However, zero order kinetics of the active substance release over a period longer than 1 day could not be observed with the previously disclosed systems.

[000401 One reason for the lack of zero order kinetics might be the insufficient average size of the nanoporous and mesoporous silica particles used, which are in the range of several hundred nanometres, as disclosed in Synthesis of Core-Shell Structured Dual-Mesoporous Silica Spheres with Tunable Pore Size and Controllable Shell Thickness", 0. Niu et at, Journal of the American Chemical Society 132 (2010), 15144-15147, and thus a too small reservoir size.

[00041] in "Mesa porous Silica Nonoparticles jbr Drug Dellvvy and Blosensing Applications", I. Slowing et al., Advanced Functional Materials vol. 17 (2007), p. 1225, the synthesis of monodlsperse separate mesoporous silica core-shell nanoparticles for a controlled-release of drugs as the active substance is described. However, In these kinds of materials the shell consists of polymers, e.g. polylactides, and Is not an entirely ordered mesoporous structure based on silicon oxide. It is obvious that these kinds of materials do not exhibit the same mechanical rigidness as materials based on an inorganic oxide framework. Since this work (partially described in US Patent No. 7563451B2) Is mainly focused on targeted delivery of drugs inside a human body and not on mere time-dependent release, there are no examples of a sustained release of a particular one of the active substance given. It is doubtful that a release over a timespan exceeding several days can be realized with the materials disclosed therein, because the maximal particle diameter of several hundreds of nanometres, and hence the reservoir size Is simply too small.

[00042] A system built of a porous rigid core and polymer shell is also described in US Patent Publication No. US2009/0304756A1, Daehne et al. The polymeric shell Is in particular adapted for a triggered release of encapsulated Ingredients by removing the encapsulated ingredients, for example by mechanical stress (see paragraphs [00621 and (0067 in US2009/0304756A1, Daehne et at). Thus, in formulations which are made for the sustained release of encapsulated Ingredients only, a sensitivity to mechanical stress resulting in an unwanted burst in the release ingredients is undesirable.

[00043] Another example for such a material exhibiting a mesoporous core and a pure polymer shell can be found in US Patent Application Publication No. U52006/0018966A1, Lin et al. Paragraph [0110J on page 13 describes in detail how the porous core is coated by a pure polymer shell, in this case made from a polylactic acid.

[000441 InternatIonal Patent Application No. W02005/009602 Is related to the afore-mentioned U5756345182, tin et al. and teaches further the synthesis of a variety of mesoporous silica particles. The use of the synthesised mesoporous particles in different applications Is described In detail. All materials disclosed are however related to a material named MCM-41, which exhibits a hexagonal 2-1) structure, e.g a hexagonal symmetry, consisting of isolated, non interconnected cylindrical channels (see page 27 line 30-32, page lIne 27-29, page 51 line 3 and line 10-12, page 56 line 15-23, page 61 fine 21-23.) or disordered, wormlike structures, also related to material MCM-41, with cylindrical pores (page 51 line 6-8, page 56 line 15-23, page 63 line 20-21). Thus, diffusion behaviour of encapsulated substances in this mesoporous silica particle cannot be regarded as isotropic.

None of the mesoporous silica particles disclosed In W02005/009602, Lln et al. show a cubic crystal symmetry with a highly interconnected channel system nor do the mesoporous sIlica particles comprise a superimposed core-shell structure. The particle assemblages taught in this document fail to exhibit a standard deviation suitable for the aimed application described In this disclosure. The standard deviation of particles can be deduced exemplarily from SEM-Picture No. 171), hF, and was determined to 7.6+-2.4 urn (31.6%) in 17D and 8.25*3.58 (43.6%) in 17F.

[00045] The mesoporous silica materials of WO 602 differ also In respect to the mean particle size and alkaline synthesis medium In contrast to the acidic synthesis medium disclosed in this current disclosure. The difference In synthesis conditions might influence the polymerization degree of the obtained silicate ("A detailed Study of Thermal, Hydrothermal, and mechanical Stabilities of a Wide Range of Surfactant Assembled Mesoporous Silica?', K.Cassiers et al, Chem. Mater. 2002, 14, 2317) and thus also influence the stability, especially the hydrothermal/thermal stability. The inventors understand that it is preferable to choose an acidic environment for the synthesis of particles. The crystal facets -12 -visible in the particles of the current disclosure might be the underlying result of such a better and stronger polymerisatlon degree, observed within our invention.

(00046) Related to the inventions described in the patent documents Nos. W02005/009602 and 1)5756345182 is US Patent Publication No. U$2006/0018966A1, Un et al. In this invention a series of mesoporous particle assemblages based on silicon dioxide is disclosed for use In different release formulations. However, none of the mesoporous particle materials out of the whole series of materials disclosed in US 966 exhibits a cubic symmetry.

On the contrary the materials disclosed in US 966 exhibit hexagonal MCM-41 type symmetries, chiral twisted hexagonal symmetry or were dIsordered (0076 to 0078 on pages 9-10, see also XRD Pattern in Fig. 4) dearly related to hexagonal structures. As in WO2005/009602 only basic media were used to prepare all of the materials and no crystal facets were observed.

(00047) In W02009/010945A2, Holmes et al, monodisperse assemblages of mesoporous particles based on silicon oxide are disclosed. The disclosed method of preparation differs however from the method of this disclosure, resulting in different materials. in W02009/010945A3 only basic media, in particular ammonia containing media are used for synthesis. This results in particle morphologies similar to the ones of W02005/009602, Un et al, but very different to the materials of this disclosure. For example, particles are by definition of W02009/010945A2 (page 10 lIne 25-26, see also claim 69) a sphere, rod, disc or rope, but not a decaoctahedron with clearly separated facets, as taught In this disclosure.

The arrangement of channels is described as being ordered in a random direction (page 9 line 27-29, page 10 line 19-20, claim 68), which excludes materials with an entirely cubic symmetry. A material with a cubic symmetry is not disclosed in W02009/010945A2.

[00048) There are several other publications describing the synthesis of mesoporous core-shell materials. However, these publications do not disclose materials that are explicitly used for sustained-release or targeted-release formulations, or the materials disclosed there are for various reasons not suitable for the aimed applications within the framework of this invention.

(00049] Entirely mesoporous core/shell particles are described in "Synthesis of Highly Monodispersed Core/Shell Mesoporous Silica Sphere? K. Yano et at, Chemistry Letters vol. (2006), 9, p. 1014, in Japanese Patent Application Abstract No JP 2006 347849A, and in "Selective Functionalization of the Outer and Inner Surfaces in Mesoporous Silica Nanoparticles", J. Kecht et at, Chemistry of Materials 20 (2008) 7207. All of the materials disclosed in the publications of K. Yano and J. Kecht possess a radially aligned pore structure of 1D channels of different length. As the channels are not Interconnected in all spatial directions, each channel can be considered as an Individual depot containing unequal amounts of the active substance, which leads to undesirable release rate fluctuations in time.

(00050] Furthermore, iP 2006347849A says In paragraph 10040] that a silica raw material is made to react in a basic solvent, sInce in acidic environments the reaction hardly advances.

This is an observation which Is contrary to the teachings of the current disclosure. The results differ also differ. The materials disclosed in JP 2006347849A are non faceted spheres, and not one of the synthesised materials exhibits a cubic symmetry. instead it Is said In paragraphs (0019] and [0020] that materials show a hexagonal diffraction pattern.

1000511 Mesoporous materials with an interconnected channel system In all three spatial dimensions are known and described in a number of examples in literature. An overview of different types of such materials can be found, for example, In US Patent No. 7767004 B2.

1000521 However, the previously described materials possess at least one major disadvantage when employed as a host material for the aimed application.

(000531 A first major disadvantage of most of the previously described materIals is the large deviation of particle size within a batch, which is not easy to overcome. This strongly limits the use of these materials as a host system. Table 1 shows some materials and their standard deviation In particle size, as well as other properties. The standard deviations were derived from disclosed SEM-pictures by counting the particles and measuring their diameter. -14 £

100054] A second major disadvantage of many of the previously described materials is severe particle aggregation that often takes place in this specified class of materials (the term "aggregation" herein is to be understood as defined by the German DIN Standard No, 53 206). The existence of separate, non-aggregated particles is a precondition for a complete, homogenous and uniform epitaxial coating of every single particle. Thus, aggregation and subsequent intergrowth is clearly to be avoided if the material Is supposed to be used as a host for the aimed purposes.

[00055] The use of bridged slioxanes In hybrid mesoporous materials, as taught in "Hybrid ethane-slioxane mesoporous materials with cubic symmetry" , Microporous and Mesoporous Materials 44-45 (2001) 165 seems to allow the synthesis of particles with a sharper size distribution, as it is shown on the disclosed SEM mlcrographs. Beside the limited diversity and unknown biocompatibility of these materials, the price of bridged siloxanes, necessary for making this type of materials, is about 20 times higher than that of typically used starting compounds such as tetraethoxysilane. This leads to extremely high, often Inacceptable costs of the end product.

[00056] Table 1 Sizet

standard SDV/Meanslze Number Publication Aggregates 100% deviation "Facile synthesis of crystal like shape mesoporous silica SB4-16"J. iln et al. 1 Yes --Microporous and Mesoporous Materials vol.97(2006), pp. 141-144 "Faceted single ciystals of mesoparous silica SBA-16 from a 4.0 ±1,3 2 ternary surfactant system: surface No 32,5% pm roughening model" B. Chen et S., Microporous and Mesa-porous -15 -Matedas voL 81 (2005) pp. 24t-249 "Synthesis of SBA-16 and SBA-l5 meso porous si/ica crystals tern plated with neutral block copoiymer 3,9 ± 1,1 3 No 28,2% surfactants" C. Lin et aL Journa of irn Physics and Chernsiry of SoUth voL 69 (2008), pp. 415-419 "Humidity sensitive property of Li-doped 30 periodic mesoporous silica 1,2±0.4 4 SBA-16" J. Tu et &. Sensors and No 333% Actuators 13 voL 136 (2009) pp. 392- "Synthesis of mesoporous silica single crystal SBA-16 assisted by fluorinated surfactants with short carbon-chains" 2,56±1,07 Some 41.7% X. Meng at aL, Micro-porous and trn Mesoporous Materiak voL 105 (2007) pp. 15-23 "Microwave synthesis of cubic mesoporous silica SBAi6" V. Hwang 6 Yes -at al, Microporous and Mesoporous Materiak voL 68 (2004) pp. 21-27 "Preparation of Highly Ordered Well-defined Single crystal Cubic Mesoporous Silica Temp/med by 7 No 6,52±1,13 17,3% Gemini Surfactant" Z. Zhang et akChemistry Letters (2002) pp. 584- (000571 Figs. 1 to 7m depict representative SEM images corresponding to the examples

given in Table 2.

[00058] Fig. 8,9, 10 depict XRD pattern, sorption isotherm and Bill pore size distribution of Example 7] of Table 2.

(00059] FIg. 11 shows a representative example of standard host material for encapsulation experiments NCap-l.

100060) Fig. 12 and Fig. 13 show SEM pictures of the achieved Core-Shell Superstructure.

(000611 Fig. 14 shows the release rate versus time of encapsulated 9-aminoacridine.

[00062] Fig. 15 shows reaction schemes for the capping procedure with dextrane.

(00063] Fig. 16 depicts several theoretical release rate of partide assemblages with different SDV calculated from Eq. 7. Mean diameter p was set to 3 a.u.

1000641 Fig. 17 shows an outline structure for the core-shell mesoporous partide of the

disclosure.

Detailed Description of the hiventlon

(00065] The Invention will now be described on the basis of the drawings. It will be understood that the embodiments and aspects of the invention described herein are only examples and do not limit the protective scope of the claims in any way. The invention is defined by the claims and their equivalents. It will be understood that features of one aspect or embodiment of the invention can be combined with a feature of a different aspect or aspects and/or embodiments of the invention.

[00066] The introduction to the application outlined a theoretical approach that serves as the basis for the invention. A practical approach involved possession of monodisperse nanoporous particles. The use of nanoporous, and especially mesoporous silica as a rigid, porous matrix for the applications is demonstrated. It is believed that cubic mesoporous materials are suitable for the application, because diffusivity in such cubic mesoporous materials is substantially isotropic and is not hindered in one or two dimensions, unlike the case of the hexagonal materials, for example, SM-is or MCM-41 mesoporous silica.

(00067] The use of a monodisperse assemblage of cubic mesoporous core-shell particles with small size deviations allows in principle the realization of formulations with the desired release kinetics. The release kinetics can follow a zero-order pattern, if the diffusivity in the core of the core-shell particle Is much higher than in the shell. it is found that the diffusivity can be tuned by different chemical modifications of the pores in the core and especially in the shell.

[00068) For many applications, relatively large particles with diameters above 1 pm are desirable. A small size of the particles in a range of up to several hundred nanometres is necessary and advantageous, when the particles are supposed to interact with, or travel through, a biological system, for example, a human body. However, the use of such small-sized materials as an inert passive depot for a sustained-release formulation is, in contemporary medical opinion, connected with an increased risk of cytotoxicity of nanoparticies, especially those with a small size, as a result of their increased ability to penetrate cells. This is discussed in "size-Dependent Cytotoxicity of Monodisperse Silica Nanoparticles In Human Endothelial Cell?, D. Napierska, Small vol. 5 (2009) 7, p. 846. It has also been reasoned that the high surface area-to-mass ratio could be an important parameter in the toxicity of the nanopartides. Thus, contemporary opinion suggests that the medical use of particles of ci mlcrometre diameter for medical applications represents significant health risks.

(00069] A further consideration is that, if a sustained-release formulation is supposed to be distributed In the environment as, e. g., a delivery depot for biocldes, particles in the nanometre range should be avoided. A particle size of 2 micrometres or more is much more -18-advantageous, because the increase of a particle size leads to a bigger reservoir capacity and thus to a prolonged timespan for the release of the specific active substance.

100070] Larger particles are also less amenable to the accumulation by lung alveoli. For example, partcles with a diameter of 2 urn are alveolar to a fraction of more than 90%, whflst particles with a diameter of 7 urn to less than 10%, as discussed in "Stacube an Arbeitsplaetzen und in der Umwe/t". M. Mattenklott et al, Stacube an Arbeitsplaetzen und in dci Luff Gefahrstoffe -Reinhaftung der Luft vol. 69(2009) 4 (April) p. 127; SpringerVDk Verlag, Duesseldorf.

[000711 This disclosure discusses the necessary host material and its modifications in order to achieve large differences in the thffusMty inside the core versus the shell and, therefore, to enable the use of the host material in sustainedrelease formulations for the active substances.

[00072] The method offers the possibility of preparing coreshe particles possessing sever shells, each of which may be different in chemical compositions. The use of such core and multi-shell particles can be advantageous, if a number of the particle's properties is desirable. Different chemical functionalities inside the particles can be used, for example, for the creation of inner diffusion-hindering layers, for making layer(s) providing a chemical modification of a released compound, a layer bearing a tag, offering better mechanical strength, or simply staining the partides with a colour, etc. [00073] The particle for use as a host for the active substance in a sustained release formulation has to exhibit the following properties: The pores should be interconnected in all three spatial directions in the same way, e.g. the material structure should belong to a cubic space group.

The material of the particle should offer a core-shell superstructure, whereas the shell should substantially hinder the diffusion of the active substances.

Diffusion of the active substances in the core should be as high as possible, optionally enabled by a liquid transport phase.

The partIcle distribution should be monodlsperse, i.e., offer a standard deviation (WV) in particle diameter generally of less than 15% of the mean particle size and in some aspects of the disclosure of less than 10% of the mean particle size.

* The size of each Individual particle should be greater than 1 pm, and In some aspects of the disclosure greater than 2.5 pm and In other aspects greater than 7pm.

* For large-scale applications, it should be possible to synthesize the material from readily available starting compounds of reasonable price, along simple synthetic routes.

[00074] The term "cubic crystal system" is to be understood as an Isometric crystal system which shows reflections In X-Ray Pattern assignable to a cubic space group.

(00075] FIg. 17 shows an example of a structure of a particle 10 that fulfills these requirements. The particle 10 comprises a plurality of interconnected pores 20 In both a core and a shell 40 and show facets 15 The interconnected pores 20 in the shell 40 have restrictions 50 within the interconnected pores 20. The partIcle 10 can be filled with an active substance. The restrictions in the shell 40 restrict elution of the active substance from the partIcle 10. It will be appreciated that further ones of the shells 40 can be added. The interconnected pores have dimensions in the nanometer range.

[00076] The properties of the core-shell particle material disclosed herein have been improved by the application of the disclosed synthetic procedures. In comparison with the methods known In the literature, for example "Control of Crystal Morphology of SM-i Mesoporous Silica", S.Che et al, Chem. Mater. 13 (2001) 2237, profound changes in the synthetic procedure and the compositions of the starting materials have been made, 1000771 It Is known that the use of two different types of surfactant Instead of a single surfactant In the synthesis of the porous particles can Influence the phase dlsperslty, shape and mean size of the obtained porous particles. This is described in "Morphology and porosity characteristics control of SBA-16 mesoporous silica. Effect of the iv! block surfoctont Piuronic $127 degradation during the synthesis", M. Mesa et al., Solid State ScIence 7(2005) 8, 990-997. The inventors have found that the simple use of two surfactants instead of a -20-single surfactant, I. e. a combination of an ionic surfactant, such as a tetraalkylammonium salt, and a non-ionic surfactant such as, for example, Pluronic F127, PElOS or F108, is Insufficient for obtaining the porous partides with the desired properties.

[00078] Addition of inorganic salts can also improve the properties of ordered mesoporous materials, as described in "Nonionic Block Copolymer Synthesis of Large-Pore Cubic Mesopomus Single Ci'ystaLs by Use of Inorganic Salts", C. Vu, I Am. Chem. Soc. vol. 124 (2002) 17, p. 4556. A simple combination of both techniques, e.g. using the co-surfactant and adding the Inorganic salt does not lead to the desired material properties for the core-shell particles disclosed herein.

f00079] The inventors have surprisingly discovered that, in order to achieve the material homogeneity, e.g., desirable size distribution and monodispersity, a co-solvent with appropriate properties has to be introduced into the reaction mixture. Additionally, the mixing order has to be altered from the mixing order common in the art.

(00080) The inventors have established that the desirable material properties are achieved when the following conditions of the synthesis are fulfilled: * A mixture of a cosolvent with a non-ionic surfactant of the poloxamer type (sub group of polyalkylene oxide), such as Pluronic F127, and a silicon source, such as TEaS, Is prepared shortly before its additIon under vigorous stirring to a mixture of the suitable Ionic surfactant and the inorganic salt, both of which are dissolved in diluted hydrochloric acid.

* The co-solvent possesses a dielectric constant of more than 30, and its structure includes a negatively polarized oxygen atom.

(00081] Table 2 &ves an overview of the obtained results in the view of the reagents mixing order, presence of a co-surfactant and various co-solvents.

* Column 1) refers to the particular example, in which the exact reaction conditions are described.

* Column 2) refers to the Figure number illustrating SEM-Pictures.

* Column 3) refers to the presence of the surfactant Pluronic F127.

Column 4) refers to the mixing order, whereas "standard" denotes the mixing order, in which afi of the reactants are mixed prior to the addition of TEDS or the particular silane or mixture of silanes, "Other" refers to another mixing order, described in detas in the examples. "Adapted" refers to a mixing order described above in the previous paragraph.

Column 5) gives the names or common abbreviations of the co-solvent used, The abbreviations denote the compounds as following: DMA: dimethyl acetamide; DMC: dimethyl carbonate; DM1: N,N-dimethyftormamide;, DM PU: 1,3-dirnethyl-3,4,5,6- tetrahydro**2(1H)*pyrimidinone; DM50: dirnethyl suftoxide; NMP: N-methyl?-pyrrolidone; THE: tetrahydrofuran.

Column 6) gives the values of the dielectric constants of the cosolvents, Column 7) gives the chemical structures of the cosolvents, Column 8) contains the mean diameter.i of obtained particle hatches, the standard deviation a in the batch, and the standard deviation related to the mean particle diameter in percent.

* Coftsmn 9) denotes, whether a strongly negatively polarized atom in the cosolvent is present (Yes or No), which in the combination with a positively polarized counterpart leads to the dielectric constant value higher than 30. In brackets, the particular atom is given.

* Column 10) contains comments on the quality of obtained particle batches, eg.

whether the particles in the batch are strongly aggregated, the obtained phase is not cubic or whether the particles homogeneity is good. 0)

C C C C. C 0 0 0 0 U 0) to 4.4 tO to 0) to to C C 0) C 0) C 0) C 0) C 0) o 00 0 tO C 0 O 0 00 is -0) a-0) to 0) a 0) t, 4.4.4_a k. 4.4 a-.-0) 44 b.-a to to to to o U to tO to to tQ L

E ii 0

4' C a-a a-n -os is w 0) to >-0)

N rn

SO

00 C + q it

I

a-n m m a, 0) I a N °1o -00 00 00 00 00 to to to 0 Cs 0 0 N N to a-fl a-0

N S

0; C.-E to a-fl 0) 0) 23 --___*o*_

H

7b 75 Yes adapted acetic add 6 H4LoH No not cubic o 2.8 ± 0.92 7c 7c Yes adapted THF 8 No arge WV (33%) 2.4±0.94 7d 7d Yes adapted 2-butanone 19 j CH3 No arge WV H3C' "N" (39%) o 2.5±0.81 7e 7e Yes adapted acetone 21 No large WV H3C CH (3 1%)

H

H 2,0 ±0.65 7t 7f Yes adapted ethano 25 Hc'tH No large WV H (33%) 3,0 ± 0.33 7g 7g Yes adapted NMP 32 N Yes [0] acceptable (11%)

CH

KG CU 3.1±0.34 7h 7h Yes adapted DMPU 36 "W""w' Yes Ed acceptable L) (11%) 2,8±0,50 71 li Yes adapted acetonitrUe 37 C CC N Yes [NJ some aggregates Hi (189 ,,,L,, *,, -L. . -24 7j 7j Yes adapted DMF 38 2.2 ± 0.26 yes acceptable (12%) 7k 7k Yes adapted DMA 39 3.3 ± 0.30 Yes (0] acceptable 71 71 Yes adapted DM50 47 246 Yes (0] acceptable H3C "CH3 (13%) 2.0 ± 0.22

Yes [0) acceptable

7m 7m Yes adapted formamide 111 (11%) -25- [0O083 A complete mechanistic explanation for the experimentally found results has not been developed. The role of the non-Ionic co-surfactant in the solution and its interactions with the additional solvent remain unclear. The necessity to mix the non-ionic co-surfactant with the co-solvent and the silicon source prior to adding the mixture of the non-ionic co-surfactant with the co-solvent to the rest of the reaction mixture cannot be explained on the base of current theory.

(00084) The dielectric constant of the additional solvent reflects the miscibility of the solvent with the silicon source, non-Ionic co-surfactant and water to a certain extent.

However, It Is not the only precondition for a successful synthesis procedure, since solvents with a Lewis-basic nitrogen atom Instead of an oxygen atom, even having a similar dielectric constant do not lead to good results (e.g. acetonitrile).

100085] With the above-described variations of the reaction conditions, it was possible to obtain regularly faceted particles with narrow size distributions and an Interconnected pore system. The partIcles exhibited a cubic crystal structure, as determined by X-ray Diffraction (XRD). Crystal morphology is described by deca Octahedron and supports the cubic system.

By comparIson with XRD-Pattern disclosed in Literature, It was found that particles belong to the cubic structure named SM-i, comprising a highly interconnects channel system. This versatile process allows for the design of complex types of the core-shell particles having multiple shells, while gill preserving the monodisperse size distribution without formation of aggregates. This approach enables a zero-order release kinetic in our release system, [00086] The variations of synthesis parameters were investigated in order to allow an epitaxial growth of optionally organically modified silica precursors avoiding secondary nucleation, and thus creating the monodisperse core-shell particles with a substantially consistent shell thickness for all the core-shell particles in a batch. Hydrolysis rate, nucleation rate, precursor concentration, available specific surface area for epitaxial growth, or precursor amount for controllable shell thIckness are only some factors that had to be considered during the synthesis.

(00087] The standard host material for the disclosed sustained release formulations comprises the particle 10 with the core 30 made of pure SiO a first shell 40 comprises free -SN groups to which, via further modifications, organic molecules or polymers are bound covalently as the restrIctions 50, and a second shell with the same composition as the core 30. The second shell was mainly Introduced to better visualize the -SH group-containing shell via SEM techniques, as described in the experImental section.

Extensive work showed that dextran with a molecular weight of ca 10000 is suitable to act as a restricting agent that decreases the diffusivity of the Incorporated active substance in the first shell of the host material. The release kinetics exhibited the theoretical predicted zero-order behavior. The use of dextran derivatives, as demonstrated in the Table 3, was surprisingly advantageous than the use of known pore blockers, such as cyclodextrins.

[00088] Table 3 illustrates the time by which half of the active substance (9-aminoacridine) is released from the depot formed by the assembly (see also description in Examples "Recording Release Curve?).

(00089] Table 3

Product No capping 6 mm Hydroxypropyl-Ø-cyclodextrln 25 mm Dextran-lO ca. 5000 mm [00090) It can be seen from Table 3 that a so-called "capping" method can be used to decrease diffusivity of the active substance in the shell 40 of the partIcle 10. This capping method involves the Introduction of organic molecules as the restrictions SO into the particle 10, optIonally in a polymeric form, that decrease the effective pore diameter of the interconnected pores 30 of the shell 40 of the particle 10 by more than 10%, and is -27 performed by covalent bonding of an organic moiety of the capping reagent to the chemically modified pore waUs in the shell. The covalent bond is optionally formed by using a cycloaddition reaction between an alkyne and an azide ("click reaction).

[000911. As a model compound for the release kinetics study, 9-aminoacridine as an active substance was used, 9-Aminoacridine is a low-toxic antiseptic that can be easily detected and quantitated due to its strong fluorescence in aqueous solutions, [000921 The particles of the disclosure have a number of potential applications. Examples are: Releasing steroid hormones such as testosteron from a parenteral polymer extrudate with incorporated hormone-containing particles in case of low testosterone levels in the body.

Releasing thyroid hormones from a partenteral polymer extrudate with incorporated hormone particles in case of hypothyroidism.

Releasing biocides such as zinc pyrithione by incorporated biocide containing particles in shoes or socks to reduce unpleasant srneil formation.

Releasing biocides such as zinc pyrithione by incorporated biocide containing particles in tubes or hoses to prevent fouling.

Releasing biocides such as zinc pyrithione by incorporated hiocide containing particles in lacquers to prevent fouling.

Releasing pesticides such as aliethrin or perrnethrin on fields; * Releasing antibiotics such as gentamicin in glues to prevent biofilrn formation.

[00093] The particles oF the disclosure can ncorporate a variety of different active substances. Non-limiting examples are summarized below.

Steroid hormones: medroxyprogesterone acetate, progesterone, estradio, norgestrel; * Peptide hormones and their analogs: leuprollde acetate, octreotide acetate, trilodothyronine; * Antipsychotics: risperidone, flupentixol, olanzaplne; * Antibiotics: gentamicin, vancomycin, tobramycin; * Antineoplastics: paclitaxel, etoposide, topotecan, cytarabine; * Immunosuppressors: rapamycin; * Non-steroid anti-inflammatory: diclofenac, nabumethone; * Analgetics: hydrornorphone, buprenorphine; * Antidiabetics: pioglitazone, gliclazide.

ExanSes (00094] The next section describes experimental protocols for synthesis of the material that fulfils the requirements for a zero-order release kinetics.

[00095] This section is divided in 6 Parts, namely * Reagents, Synthesis of raw materials * Analysing Techniques * Synthesis of Host Materials * Loading of Host Materials with AS * Capping a Release Experiments Reagents, Synthesis of raw materials (00096) Unless otherwise noted, all chemicals were purchased from Sigma-Aldrich in reagent grades. Reactions were performed at room temperature. Water was delonised.

(00097] Cetyltriethylommonium bromide: Into a 2 1 round-bottom flask was placed hexadecyl bromide (250 g), 2-methoxypropanol (250 ml), and triethylamine (200 ml.). The flask was heated without stirring at 75 C for 96 h and the contents were concentrated on a Rotavap at 75 C and 20 mbar. To the residue, methyl tert-butyl ether (800 ml) was added. The slurry was vigorously stirred by a mechanical stirrer for 12 h, filtered, washed with 3 portions (400 mL each) of methyl test-butyl ether, and dried in vacuum to obtain the product as a white solid In nearly quantitative yield.

(00098] 9-Aminoocrldine (base): 9-Aminoacridlne hydrochloride monohydrate (2.5 g) was mixed with 5% aq. NH3 (25 ml), stirred for 3 h, filtered, washed with water (30 ml), ThF (30 ml), and dried In vacuum.

100099] 4-Azidobutyeyldextran-1O solution: In a 100 ml round-bottom flask under argon, 1,1'-carbonyldilmldazoi (3.0 g) was dissolved In dry DM50 (25 ml), and 4-azidobutyric acid (1.5 g) was added. After 3 h, dry dextran-lO (4.5 8) was added, and the mixture was heated under argon at 75°C for 18 h. On cooling, ethanol (300 ml) was added to the mixture. The oily precipItate was washed 4 times with boiling ethanol (100 ml portions), and dried at 60,C. The residual solid was dissolved in water to obtain a 10% solution that was filtered using 0.22 m syringe filter and used without further purification.

(000100] 4-Azidobutyrl-hydroxypropyl-8-cydo4extrln; In a 250 mL round-bottom flask, dicyclohexylcarbodlimide (6.0 g) was dissolved in dry DMF (30 ml), and 4-dlmethylaminopyridine (3.4 g) was added. To this solution, hydroxypropyl4-cyclodextrine (8.0 & Aldrich, average Mw = 1460 Da) was added, and the mixture was stirred for 5 days at ambient temperature. Solvent was removed in vacuum, the residue separated between cii2a2 (50 ml) and deionized water (100 ml), aqueous layer extracted twice with CH2CI2 (50 ml), and passed through a column containing ion-exchange resins: 35 g of Amberlyst 15 in H' -form and 35 g of Amberlyst A26 in 0H form. The column was washed with 100 ml of deionized water, the solutions were combined, concentrated in vacuum to ca. 25 mL, and lyophilized. The product was obtained as colorless foam.

Analysing Techniques 10001011 Sorption isotherms were recorded with a Quantachrome NOVAe using nitrogen at 77 K. Samples were degassed for 12 h in vacuum at 393 K prior to measurement. Surface -30 -area was determined via the BET-Algorithm, pore size distribution via BJH-Aigorithm applied to the desorption branch.

(000102] Powder-X-ray diffraction was recorded on a Bruker D8 using Cu-1d radiation and 0.07 2-theta steps.

1000103] Scanning electron mlcrographs were recorded with a Phenom 61 from Phenom-World By.

(000104] Particle size distribution was determined from SEM Images using the software Imagei.

(000105] The core-shell superstructures were made visible in SEM-Micrographs by partly breaking the particles and Improving the contrast between the cores and the shells by increase of the electron density through the binding of gold(llI) ions to 511-groups in the shell(s). Samples were partly destroyed by gentle grinding. ApproxImately 20 mg of the gently ground powder was stirred for 20 minutes in 5 ml of 1.5 mM aqueous AuCl3, and washed thoroughly with water and acetone on a Buchner funnel. After drying on the Buchner funnel, SEM pictures were Immediately recorded as soon as the sample exhibited a slightly yellow colour.

(0001061 The concentrations of the released substances were determined by calibration curves after measuring the fluorescence of diluted aliquots using a Hoefer DynaQuant 200 Fluorometer.

Synthesis of Host Materials [000107] For all experiments, a Stock Solution 51 was prepared by dissolving 11.54 g of cetyltriethylammonlum bromide and 141 g of sodium sulphate in 1190 mL of 3.36 M aqueous hydrochloric acid. The solution was stored overnight at ambient conditions prior to Its first use. Stock Solution 52 was prepared by dissolvIng 10 g of Pluronic F127 In 100 g of N,N-dimethylformamlde.

[000108] Examples evaluating the influence of rosa/vents1 Pluranic F127 template and mixing order.

[0001091 A experiments were carded out by mixing 20 g of Solution Si with optional additives. This solution was named Al. To the prepared Al solution, a mxture named A?, comprising 0.3 g TEOS and optional additives was added, stirred for 20 seconds, and stored for 30 minutes on the bench. The mixture was transferred to a rotary shaker and stirred with 1 RPM for 90 minutes, fiftered, the soiid washed with water and dried. Table 4 displays the compositions of Al and AL The number of examples refers to numbers in the first column in

Table 2

[000110) Table 4

Example Al A2

I 2OgSl 0.3 gTEOS 2 2OgS 0.1 g F127 O,3gTEOS 3 20 g Si, 0,lg F127, 1 g DMF 0.3 g TEOS 4 20 g 51, 0.1 g F127 1 g DMF, O.3g 1105 S 2OgSl4gDMF 0.SgTEOS 6 2001 IgDMF,0.3gTEOS 7am 20 51 1g cosolvent, 0 ig F127, 0.3 g TEOS [000111] Examples 7aJrn were carried out simiarEy, however, the co-solvent used was the solvent listed in Tabie 2.

[000112] The reaction mixture in example 7j was heated in an autoclave for 2 hours at 120°C. the white suspension was fiftered off by a Buchner tunnel, and placed in a bottle wfth -32 -ml of the mixture of 10% wt. conc. HCI and 90% wt. of ethanol. This treatment was repeated twice, then the solid was washed with lsopropanol and dried at 90t.

(000113] The XRD pattern of this material is displayed in Fig. 8, the sorption isotherm in Fig. 9 and pore size distribution In Fig. 10. The XRD reflections were assigned to a cubic structure, with reflections (200] at 2.02 2$, (210) at 2.25 °20 and [211] at 2.465 020 [000114] in all other experiments the phase purity was checked via XRD measurements without template extraction.

(000115) Standard host material used for release experiments.

Synthesis of the core [000116] 3.3 g of SolutIon 52 was mixed with 0.9 g of tetraethoxysllane, the obtained clear solution was poured Into 60 g of Solution Si, stirred vigorously for 20 seconds. This mixture was named "Reactant Solution 1 (Ri). The solution was stored at ambient conditions of 30 minutes and then put into a rotary shaker (1 RPM) for another 30 mInutes in a plastic vessel, offering a total volume of ca. 75 ml.

[000117] The second Reactant Solution (R2) was prepared 1 hour after having started with RI, In the same manner. However, all the reagents quantities were multiplied by a factor of 5. Additionally, after mixing Si, TEOS and 52, the previously prepared solution Ri was added, and 340 g of the final mixture were immediately placed in a rotary shaker for 2 hours and 15 minutes at 1 RPM. The vessel used had a total volume of ca. 350 ml.

Synthesis of first shell containing SN functional groups [000118] A reactant solution R3 similar to Ri and R2 was prepared 2 hours and 15 minutes after having started preparing solution R2. The reagents quantities were multiplied by a factor of 5.7 of the ones used to prepare Ri, and TEOS was replaced by a mixture of 95% wt.

of TEOS and 5% wt. of 3-mercaptopropyltriethoxysilane. This solution was added to 340 g of -33 the mixture consisting of Ri and R2, and placed in a rotary shaker for 3 hours at 1 RPM, in a vessel with the volume of Ca. 102% of the total volume of the liquids. The 3-mercaptopropyltriethoxysilane is responsible in this example for the creation of the -SH functional groups.

Synthesis of a second shell with pure TEOS as a si//con source [000119] A reactant solution R4. similar to Ri, was prepared 3 hours after having started the preparation of R3. The reagents quantities were multiplied by a factor of 83, The previously prepared mixture containing solutions Ri, RI and R3 (ca. 700 g) were added to R4. The resulting suspension was kept in a rotary shaker for 2 hours at 1 RPM in a vessel of the volume of ca. 102% of the total volume of the liquids. Two hours after having started preparing Solution R4, the suspension was ffltered, the white sohd was washed from the fHter into a glass bathe using Ca. 200 ml of diluted hydrochloric acid, and the dosed bottle was placed in an oven at 90t overnight.

Template removal [000120] The white suspension was filtered off on a Buchner funnel and placed in a bottle with 200 ml of a mixture of 10% wt. conc, HCI and 90% wt. of ethanol for several hours in a shaker. This treatment was repeated twice, then the solids were washed with isopropanol and dried at Sot E000121J The sample so obtained consisted of pure SBA-1 mesoporous silica with a cubic structure, as was determined by XRD, and exhibited a surface area of 1223 m2/g with the pore size diameter of 2.2 nm. The mean diameter of particles was 636 ± 0.61 pm (9%). A SEM-Picture is displayed in Fig. 11 in which the cubic crystal morphology of decaoctahedrons is clearly visibly. ASEM Picture of sheIk is depicted in Fig. 12 and Fig. 13. This material was named NCap4.

Propargylation ofSH Groups -34 - [000122] 3,3 g of NCap4, bearing free SEt-groups in one of the sheDs, was added to a mixture of 2-rnethoxypropanol (15 ml), dsopropyethylamine (1,2 ml), and propargyl bromide (1 ml of 80% toluene solution). This suspension was allowed to react in a shaker for 24 h, fiRerS, washed with methanol and dried in vacuum.

Loading of Host Material with Active Substance 1000123] 03 g of a 15% solution of 9-aminoacridine hydrochloride hydrate in OME was dropped on 1 g of the propargylated NCap4. The powder was shaken in a 10 ml round bottom flask for 3 minutes and checked afterwards, whether any clustering took place. If so, the clusters were carefully ground with a spatuia, and the powder was shaken for further 3 minutes. This process was repeated, until no more clustering was observed, The powder was dried in an oven at 90t for 2 days in an open vessel.

Capping [000124] apping with Dextrnn-iO Propargylated, 9-arninoacridine!oaded NCap4 (1,0 g) was mixed with 10% sohjtion of 4 azidobutyryldextran4O (2 ml), and the sodium ascorbate solution, prepared from Lascorbic add (400 mg), and NaHCO3 (200 mg) in water (2 ml). A 7% aq. solution of CuSO4 (0,3 ml) was added, and the resulted suspension was placed in a shaker for 72 h at ambient temperature. The sodium ascorbate solution is used to reduce the CuSO4 to Cif ions which can catalyse a cycloaddition reaction. Water (5 ml) was adds, and the suspension was centrifuged. Washing with 5 ml portions of water and centrifugation was repeated until the amount of washed out substances was negligible. The obtained yellow solid was dried in vacuum.

[000125) Capping with hydroxypropyl4Gyciadextrin The capping was performed according to the procedure for the capping with dextran, but 10% solution of $azidobutyryl-hydroxypropyl4-cyclodextrin was used instead of the corresponding dextran derivative, Recording Release Curves [000126] 50 mg of the products obtained as descdbed above (dextran**capped hydroxypropyl-cydodextrin-capped, or uncapped NCap-*1, loaded with 9aminoacridine hydrochloride) was washed thoroughly placed into Ca, cm piece of a dialysis membrane (MWCO 1000044000). and immersed into 200 ml of water in a plastic beaker, The plastic beaker was fixed in shaker. For concentration measurements, aiquots (50 á) were taken after certain periods of time and diluted with water (SM ml). Fluorescence of this solution was measured (excitation 365 nm, emission 460 nm). The concentration of the released in the test vessel 9aminoacridine was determined using a calibration curve. The Urne by which a half of the substance was released from the sample was calculated, The results are given in

Table 3.

[000127] From the obtained concentrations the release of the active substance per time was calculated. The dependency of the release rate on time for the dextran..capped NCap4 is depicted in Fig. 14.

Claims (6)

1. CIa ms 1. A substantially monodisperse assemblage of particles 10 having interconnected nano sized pores 20 and a core 30 with a shell 40 disposed about the core 30 and wherein the partides 10 have a cubic crystal form.
2. 2. The assemblage of particles 10 of claim 1, wherein the particles are selected from the group consisting of metal oxides and metalloid oxides, preferably from silicon oxide.
3. 3. The assemblage of claim 1 or 2, wherein the average particle size of the particles 10k greater than 1 micrometres, more preferably greater than 2 micrometer, more preferably greater than 6 micrometer,
4. 4. The assemblage of any of the above claims, wherein the standard deviation of the particle size is less than 15%, more preferably less than 10% of the average particle size.
5. 5. The assemblage of any of the above claims wherein the particles 10 have a Faceted surface 15.
6. 6. The assemblage of any of the above claims, wherein the interconnected nano*sized pores 30 of the shell 40 comprise restrictions 50.7... The assemblage of claim 6, wherein the restrictions 50 comprise organic molecules or polymers covalently bound to an inner surface of the interconnected nanosized pores of the shell 40.8. The assemblage of any of the above claims further comprising an active substance in at east some of the interconnected nanosized pores 20 of the core 30.9. A method for the sustained release of an active substance to an environment comprising: -adding the active substance to a substantiafly monodisperse assembage of particles having interconnected nano-sized pores 20 and a core 30 with a shell 40 disposed about the core 30; placing restrictions 50 in the interconnected nano-sized pores 20 within the shell 40 by covalent bonding; and placing the substantiaUy monocfisperse assemblage of partides with the added active substance in the. environment.10. The method of claim 9, wherein the active substance is an active pharmaceutical ingredient.11, A method for the manufacture of a plurality of monodisperse particles 1.0 comprising: -mixing an ionic surfactant with an inorganic salt in hydrochloric acid; -mixing a co-solvent with a first non-ionic surfactant and a first oxide source; -mixing the solution of the ionic surfactant arid the norgank salt with the solution of the first non-ionic surfactant and the first silicon source in the first co-solvent; 12. The method ci claim 11, wherein the oxide source is a silane.13. The method of claim 11 or 12, wherein the co-solvent is selected from the group consisting of dimethyl acetamide, N,N-dimethyftormamide, 1,3-dirnethy3,4,56-tetrahydro-2(1H)-pyrimidinone, dimethyl sulfoxide, N-methyl--2.-pyrrofldone.14. [he method of any one of claims ilto 13, wherein the non-ionic surfactant is a polyalkyleneoxide, 15. The method of any one of ciaims 11 to 14, further comprising: -mixing a second co--solvent with a second non ionic surfactant and a second silicon source; -adding after a period of time the solution of the second co-solvent with the second non-ionic surfactant and the second silicon source to the sohition of the ionic surfactant and the inorganic salt and the first non-ionic surfactant and the first silicon -38-source in the first co-solvent.16. The method of any one of claims 11 to 15, further comprising functionalising inner walls of at least some of pores in the monodlsperse particles 10.17... The method of any of claIms 11 to 16, further comprising adding an active substance to the monodisperse particles 10.18. The method of claim 16 or 17, further comprising attaching restrictions to the functionalised Inner walls.19. The method of claim 18, wherein the restrictions are formed of dextran polymers or derivatives thereof.