SE527505C2 - Composite materials and particles - Google Patents

Abstract

The invention is a method of making a composite material (from a route given by the phase behavior of a suitable chemical system that is described in a phase diagram), which comprises at least one amphifilic component and at least one polymer component. It comprises providing a mixture of at least one polymer and at least one amphifilic compound in a volatile solvent or solvent mixture as well as providing a phase diagram of the chemical system that describes how the components of the chemical system interact in thermodynamic stable phases as a function temperature, concentration and pressure. The polymer should be a homopolymer, a random block copolymer or a mixture thereof, preferably biodegradable. The amphifilic compound has the ability to form a bilayer-containing phase. The solvent is removed from the mixture by a process selected from the phase diagram in dependence of the final composite material to be achieved, whereby a material is formed, such as liquid extraction against a second solvent, or by spraying. It also relates to the composite material, in particular particles, solid implants, semi-solid, gel-like matrices, useful for applications such as encapsulation of therapeutically active components or surface coating.

Description

65 70 80 527 505 2 The lipid bilayers, cubic phase, are connected in three dimensions (BD) and separate two separate systems of water channels. The size of the water channels (diameter) varies with the composition of the formulation (50-100 Å).

The cubic phase is particularly interesting as a candidate for applications, where the biologically active substance is to be under- inserted over a long period of time, so-called controlled (delayed, retained) release or delivered. The incorporated substances are, in vitro, expected to be inserted according to the "zero order kinetics" (proportional to the square root of time).

Unsustainable attempts to find suitable systems for the formation of lipid / polymer composite particles have previously been performed and reported (Journal of Physical Chemistry B 2001, 105, 12157-12164). In the study, 1-methyl-Z-pyrrolidinone (NMP) was used as solvent E ersom because NMP is distributed to the water to a very large extent, the polymer phase becomes extremely concentrated, which means that particle formation, by classical emulsion method, is not practically unresponsive to the quaternary referred to above ( 4 component) system.

The following patents and patent applications are cited herein because they involve the simultaneous use of the same lipid and the same polymer as in the denim invention (with other uses for the surface crystalline axis): U.S. Pat. No. 6,277,4l3: by Sankaram et al., Describes how encapsulated substances can be controlled by varying the lipid / polymer composition into lipid / polymer composite particles.

US pat. No. No. 6,488,952: by Kennedy et al., Describes how an multiparticulate system, for injection, depot or implantation, can be formed by incorporating polymeric microparticles into a "hydrogel".

International patent application, reference number WO 2002/19988 A2, by Bodrneier et al., Describes a method for preparing polymer microparticles, with great potential for varying the manufacturing conditions, by using a emulsifier (M0). International Patent Application, Reference No. WO 99/47588, by Dawson et al., discloses a method for preparing polymeric microparticles, over a wide range of preparation conditions, by using MO as the emulsifier.

Summary of the Invention Consequently, the object of the invention is to provide and protect; (i) a suitable system, i.e. a group of grain components, for the formation of composite materials, in particular lipid / polymer composite particles with roller-separated lipid domain (s), for example dispersed (e), in a biodegradable polymer matrix, for delivery of sensitive biologically active components and (ir ') a suitable manufacturing method for such composite materials, eg lipid / polymer composite particles. The lipiral domains should preferably swell and form a cubic (or other [liquid] crystalline) phase in accordance with the phase diagram describing binary lipid (or other surfactant component / water system in aqueous environments).

The sewing is achieved by using a suitable system with suitable behavior which results in a noble manufacturing path for lipid / polymer composite materials, such as particles, implants, or for applications in, for example, surface treatment, cosmetics. Critical foods, veterinary medicine, separation techniques (ion exchange pads) and applications in farnnaceutsik technology, such as dachshund granulation and tabletting.

The invention is based on the use of a volatile solvent, which is not completely miscible with water. The solubility of the solvent in water is 8% (at 20 degrees C). Because the solvent is not always miscible with the water, it is suitable for use in particle formation by any emulsion method. The partial miscibility of the solvent with water has been used to control / control the genetics of lipid domain formation, making it difficult to study the phase separation by conical laser scanning microscopy.

The invention comprises a suitable system (combination of components) which gives rise to a desirable etching behavior (with specific and general properties) which enables the formation of such material and particles. Compositions for composite materials in general and particulate matter in particular, according to the invention, are claimed and based on the method of emulsification; the appropriate amount of polymer / lipid / solvent ms (s) is homogenized into a liquid phase (which may be water - but which may also be another solvent / solvent mixture) as the intrinsic solvent solvent polymer, i.e. Other methods can also be used; eg spray drying, atomization and methods based on super-critical "id uid" telcník. Other, traditional, chemical and physical, microencapsulation methods of interest are; spray cooling, fl unidisised bed techniques, extraction, spruce surface polymerization (depending on choice of polymer) etc.

Internally, the invention is presented with a selected concentration range presented by Lin, which material and / or particles can be formed by a controlled “removal” of solvent.

In particular, it should be noted that the invention uses lipid's (or other surfactant molecules) that self-associate functionality, ie its ability to swell and self-associate in mono- or bilayer-containing structures (phases), for e.g. applications such as delivery of active substances .

The water channels, present in the cubic axis, are used as an encapsulation of hydrophilic substances. Alternatively, the membranes formed, here consisting of annex structure, can be used for encapsulation of hydrophobic substances. Thus, the system can be used in “combo products”, ie containing your therapeutically interesting substances located in the hydro or hydrophobic domains.

In accordance with the invention, formed appendages (membranes), which form the cubic phase, can be used for the protection and encapsulation of membrane-bound proteins as well as other passive eam lataapeu fi sktaküvas subsmnsenmvissa fi serbestårravmomhger 150 155 160 165 170 175 180 185 190 195 527 505 L: these can also be used for the incorporation of surfactant relevant components.

Intrwantärocksåattlipidenarpassandetbrattøkformirleringens _ _ vedhäftningstörmög till en slemhinna, the so-called "mucoadhesione fi ï Formulermg fifl, 1 agreement with the invention, is interesting in applications such as exernpelvrsfgyrmat uptake and transport of particles, increased vaccination and vaccination and immunization.

According to the invention, the formulation can be used for applications such as controlled or delayed kinetics, the dry separation between the lipid and the polymer is adjusted by using different molecular weights and types of components and renodications of the grain components (eg rhododised groups on the lipid and polymer).

In accordance with the invention, it is possible to use any polar lipid (or other desirable (surfactant) molecule) with the ability to form desired (surface crystalline) axes in aqueous environments. Examples of such polar lipids are monoelaidine, phosphatidyletharrolamine, phospholipids and polyethylene glycol (PEG) -modified phospholipids.

The lipid / polymer composite particles must contain at least one biological polymer. The polymer may be a homopolymer (copolymer), a random block copolymer, or a mixture thereof. The polymer must be partially or completely soluble in organic solvents and not completely soluble in water.

Examples of biodegradable polymers useful for the formation of lipid / polymer composite materials and composite particles are, for example; polyactide), 1> ° l> '(a1>' k ° lid °), POMP-dißx fl wne), pol fl caprvlßßtßne), wlyhydr hydr xyalkanoate, polypropylene mar rmarate, polyorthoesters, polyphosphate esters and polyarrhydrides.

The lipid / polymer composite particles are suitably formed with lipid domains dispersed in the polymer matrix, or with a lipid (or polymeric) core within a polymer (or lipid) shell. As indicated above, in addition to particles, solid implants and semi-solid, gel-like matrices or hydnogels are formed. By using water-insoluble polymers the former formulations (solid) can be formed and by using slightly more water-soluble (ie hydro-1-modified) polymers the last type of formulations (semi-solid, swelling, gel-like) can be formed.

From a specified aspect of the invention, very porous (polymer or lipid / polymer) particles can also be formed in the chemical system which is reported in accordance with the invention. Deporose particles can be formed according to the elution two-step process: In the first step, lipid / polymer composite particles are formed according to the detailed description given in Example 1. In the second step, the water-swollen lipid domains are washed out of the polymer matrix with a solvent that dissolves the lipid but not the polymer. In this way, very porous polymer particles can be created. Such particulate matter may be associated with thermoplastic agents, be 200 205 210 215 220 225 230 235 240 245 527 505 5 interesting inhalation applications. The final degree of porosity depends on the amount of lipid (or other component present) present in the formulation (as well as which degree of component component is allowed to swell in water sub-stage 1). It is reasonable to measure porosities of 5-70% by varying the lipid content and degree of water swelling (amount of water present and the time in the water-rich environment). The possibility of being able to vary the pomsity in such a range is a big advantage of the system.

Another application of the invention is to use the system of debranules. The genome uses a strong phase separation and allows water to separate the lipid into the surface of the particles. The phenomenon can be used in Stearate is well used as a binder / adhesive, but also the above-mentioned lipids (or other surfactants) can be used. Important properties are then to choose the right strength for the separation of the desired bar surface of granulate material. The benefit is that granules can be produced in a one-step process.

An interesting application of the system, in the field of multidimensional food, is the possibility of delivering innmktinnella (essential) lipids such as omega 3 and 6.

Another interesting application is the use of tissue tissue regeneration technology, bone regeneration, bone regrowth and stem cell application, as well as important factors, including the subdivision of hydro and localized hydrocarbons.

Brief Description of the Figures Figure 1. The phase diagram describing the behavior of the quaternary lipid / polymer / solvent / water system (at 20 degrees Celsius).

Figure 2. A schematic representation of the cubic axis.

Figure 3. A schematic description of the manufacturing method.

Figure 4. Confocal laser scanning probe microscopy, CLSM, image taken after 23-26 minutes after manufacture of MO (lipid) / PEG-PLG (polymer) composite particles, manufactured with, 6% EtAcidenyttreiàsen.

Figure 5. A CLSM image 23-30 minutes after manufacture, of PEG-PLG polymer (reference) particles, made with, initially, 6% EtAc in the outer axis.

Detailed description of the invention The description and examples below illustrate the invention in detail. However, the chemical system illustrated in the examples is only an example of a suitable choice of grain components, i.e. a method in accordance with the invention. Most variants exist, within the framework of the invention as they are initiated in the requirements, and for people in the field, it is possible to use the invention to forecast / evaluate related systems and materials. Although the reference in the examples only concerns particle formation, it should be pointed out that the invention has a much wider applicability than that, and that it is possible to manufacture implants as well as semi-solid, gel-like matrices / implants, etc. by using it in the invention. described technology. 250 255 260 265 270 275 280 285 290 295 527 505 b Solvents here refer to both simple components such as ethanol, cyclohexane, ethyl lactate, ethyl acetate, or water and mixtures of one or more solvents.

The figure is a schematic illustration of the near phase diagram showing the interest in accordance with the reference, in particular to the assimilation of the cubic axis. The snuktrrren of the cubic phase, i.e. its water channels and appendices are further presented in the General Description of the base which is partially read on the quarter of the poly (d, l-lsctide-co-glyeolide) (PLG) / rnonoolein (Et) and water / ethyl acetate.

The complete avter quarter type system clamping of entetrades. Figure 1 shows the diagram of the ternary systems EtAe / MO / water, EtAc / M0 / PEG-PLG and MO / PEG-PLG / water. Although no systematic study of "insidanavensádan fi stenaede fi harrntönskanândåen al lrhå al andevísbra al lmtáelseavíàbeteendetavdetkvmternämsystemetahá al asgemmattstudem läsbeteendenaíundersystemen, vilkabeskriversidornailästetraedern, somalla explained further below Note that the type of PLG used Sat beståmningen of 'rasdiagrammm in fi gure 1, the PEG segments incorporated in polyOactide- eo -glycolide) backbone. Qualitatively, the phase behavior is the true pure poly (d, l-lactide-co-glycolide) polymer used. However, the heat EtAc concentrations, i.e. the amount of EtAc needed to form a true one-phase solution of MO and PLG in the EtAc / MO / PLG system, vary with specific properties, such as type and molecular weight. Tvolic polymers have nanoparticles. The first is a classic PLG and the other, recently developed modal PLG, has PEG segments connected to the PLG plain. The purpose of using PEG-modified PLG is that PEG segments can be present only at the interface surface between MO and PLG, and thereby lower the boundary surface tension (surface energy. In this case, the PEGylation is also used to stabilize, for example, sterile aqueous or polymeric lipid). -on the polymer matrix.

FasbetændetvisarattMOoehPLGsepMmariWáoIikafaæ fl dVsdehaIenNpNsíV interaction between themselves). The phase separation can, as the name implies, be reduced by the addition of suitable solvents (eg EtAo) which further dilute the unfavorable interactions between M0 and PLG. Thus, at high EtAc concentrations, the M0 and PLGs may coexist at the same time as the common phase, which may be called the phase. If EtAc is slightly soluble in water, the phase separation between MO / PLG of water addition is indicated (ie EtAc is transported from the L-phase to the water).

For applications such as the formation of particles with lipid domains, the solvent-rich single-phase solution, L, can be used as a "starting solution" which will be further described below. Emulsion droplets (which later become particles) are formed by homogenizing the starting mixture (L-ion) in an outer aqueous solution. As a result, EtAedi fl indents to the new water and dissolves, making lock separation of M0 and PLG. During the separation, MO domains are formed in the PLG matrix. When the EtAc content of the enrulsion clap is sufficiently low, the polymer phase is properly concentrated 300 305 310 315 320 325 330 335 340 345 527 505 i * (which means that the polymer matrix is hard and particles are formed in the emulsion droplets). A more detailed description of the particle welding process is given below.

In applications, it is important to check the kinetics of the phase separation between MO and PLG. Different properties, which can be varied, affect the strength of the lirse separation.

For example, the ligase separation, also called segregation, is affected by the properties of the polymer, such as molecular weight, degree of PEGylation and length of PEG. High molecular weight and low pegylation (proportion and length) result in a strong lice separation. PEG probably lowers the surface energy between the lipid and the polymer and thereby affects the kinetics, (strength - driving force) of the phase separation. To adjust / control the phase release between lipid and polymer, it is possible to use different lengths of the PEG chains as well as different degrees of PEGylation. In general, the tendency for lipid-polymer separation to decrease with increasing degree of pegylation decreases.

The pure poly (d, l-lactide-co-glyeolide) polymer, purchased from Boehringer Ingelheim Pharma GmbH a co Fine cnemicars (nrgemeim am Rhein, Germany). has a zero molecular weight of 50,000 calculated as a rest average value, which corresponds to a molecular weight of 11,000 calculated as a number average part value.

The pegylated PLG2 obtained from Birmingham Polymers Inc. consists of 70% PLG and 30% PEG 1000 (lot no. D99164). The inherent viscosity is, according to the manufacturer, 0.45 dl / g.

The phase behavior of the EtAc / MO / PLG system The phase separation between MO and PLG can be reduced by adding a suitable solvent, such as EtAc. Where fl ir is PLG and M0 miscible, in an L phase, mr EtAc content is high. EtAc is better distributed to M0 than to PLG and the system is an example of a so-called. asymmetric system. The critical EtAc concentration levels, ie the lowest EtAc concentrations required to form the readings (at all M0 / PLG content groups) are 99% in the case of lower PLG and 88% in the case of the PEG-modified polymer.

Phase behavior of the EtAcJPLG / water system Almost all water forms an EtAc / water mixture that coexists with a PLG-rich phase, see Figure 1. The slope of the binding lines, schernatically illustrated in Figure 1, indicates how the polymer-rich phase is concentrated during particle formation (as described below ).

The phase behavior of the EtAc / MO / water system The phase diagram of the ter-close EtAc / MO / water system, presented in Figure 1, contains the following single-phase regions; (1) a liquid keir (L-lirs), (ii) two different bicontinuous cubic phases, namely the so-called. The Gyroid and Diamond structures and (iii) a lamellar (or crystalline) region. Between each single-phase region presented in Figure 1, there are two and three regions. 350 355 360 365 370 375 380 385 390 395 527 505 It is noteworthy that the water content in the L-phase increases at approximately 35% EtAc, which is most likely to lead to poor aggregation (self-association - which may be due to the MO transition to the surface / action temperature).

Practical Particle Formation The Ernulsünslvretoden Within the framework of the invention, particles can be formed by different methods.

The emulsion method is an example of a possible way to remove the solvent: Lipid / polymer composite particles are formed according to the following procedure, which is further described in Example 1, based on the emulsification of a liquid phase (L phase) into an aqueous kingdom.

Now the formation of the L-anen Lipidpmbnn, Linsnnninn W rhodamine B 1 Z fi hexadecanoyl-sn-glyeero-S-phosphoethanolamine triethylamminnium salt (rhodaniine DHPE), is first dissolved in 95% ethanol and then mixed with MO. The lipid / probe solution is incubated at 50 degrees C until the ethanol has evaporated. Polymer and solvent are added there so that only a true solution, the L-phase (see Figure 1), is obtained.

The L-ice used for the formation of reference particles contains the same amount of DHPE probe, PLG and EtAc as the L-phase used for the formation of lipid / polymer composite particles. The only difference is that condensers also contain some MO.

Particles are formed genomically by emulsifying 0.1 mlL-fi as (the composition of L-phase is given in Example 1) in 1.16 grams of the outer, water-rich, phase consisting of 0%, 4%, 6%, or 8% EtAc (in water). In both phases, the room temperature emulsion is formed with a polytron mixer kinematica (P'I'300) which operates for 3 minutes at 4000 rpm.

Description of the Particle Formation from the Phase Behavior In the phase diagram, which is presented in Figure 1, schematic binding lines are given. Binder lines reveal that both the PLG / EtAc and MO / EtAc mixtures can coexist (equilibrate with) an EtAe / water mixture. The slope of the binder line indicates that a “stable” ernulsion, consisting of droplets of the Iffase dispersed in the outer water ice, is easily present in this system. Thus, the system is suitably used for the particle formation of lipid / polymer granule positive particles by an emulsion method.

Solid particles are formed when the polymer in the emulsion droplets of the L -ase is concentrated / precipitated as a consequence of simultaneous removal of the solvent of the polymer. The removal of the solvent, from the emulsion drop, takes place because the solvent has a certain miscibility in water. It follows that the solvent will immediately disintegrate out of the outer phase as a small amount of L-phase emulsification external fi-standing effluent. The co-solvent 400 405 410 415 420 425 430 435 445 527 505 The soluble solvent evaporates from the system, which means that more solvent dimbs out of the droplets to the outer aqueous phase. The polymer axis is concentrated, which means that solid, hazardous particles are formed.

At the same time as the polymerase is concentrated, the lipid phase separates from the polymer phase and forms lipid donors inside a polymer matrix. When water enters the polymeric mats, the lipid domains swell according to iden gnr LUnder fl irreplacement to lipidârisitt fl superficial stateblnnersiglipidenßrstienvâtske fis, L-phase, according to fi gtn '1. As a result of further water swelling, ie further swelling in lipidation, i.e. Ir lamellar (IQ) - cubic G (QG) -cubic D (QD). Densistnämndaktrbiskaiiasensamexisterariöverskounndæmvattenß fl er envattenríkEtAc / vanniäs), according to figure 1. Semre, låmnardevattensvällta lipidomânerna polymer-malrisen. PLG / MO composite particles emit pure water-swollen lipid domains and PEG-PLG / M0 composite particles emit water-swollen lipid domains having a polymeric shell (PEG) around them. The fact that the latter lipid donors are surrounded by a polymer cell is probably due to the fact that PEG, provided that the PEG chains are not too long, is miscible with water.

Important properties of the chemical system One of the most important properties of the chemical system, from the point of view of particle formation, is that the L-phase, according to Figure 1, can exist in an ethnodynamic equilibrium below the EtAc / water mixture. The biphasic compound exists in the manufacture of composite particles in addition to the learning of emulsions described above.

The system is thus stable and easy to use for applications such as particle formation outside, for example, emulsion methods. Schematic hyphens, illustrate in the EtAe / PLG / water system which two phases are the state-of-the-art composition. The slope of the binder lines illustrates how the PLG / EtAe phase changes its composition during the particle formation process, ie when the solvent leaves the system by evaporation.

Another suitable property of the chemical system is that the soluble solvent, only to some extent, is miscible with water. This property makes it possible to control and study the kinetics, of the lipid / polymerase separation, by varying the EtAc content in the outer aqueous phase then. the emulsion is created.

The kinetics reflect the rate of transport of the EtAc å from the emulsion droplets / particles.

Yet another suitable feature of the system, which also affects the kinetics of the lipid / polymer ice separation, is that the lipid may begin to swell, i.e. incorporate water, while EtAe is still present. The overall kinetics of lipid / polymer phase separation depend on both; (i) how fast the lipid is rising (ii) how fast the EtAc leaves the emulsion drop.

Another very fitting feature of the system is that the cubic fi tsenkaneexisted with certainEtAc content. Here, the encapsulation of therapeutically active active substances in the lipid domain can be expected to result in a single attachment, bending down according to the cubic phases, during most of the particle formation processes (when emulsion methods are used). 450 455 465 470 475 480 485 490 495 527 505 10 In summary, from the point of view of particle formation, the system is suitable for use.

An important property of the system is that the lipid can begin to swell (incorporate water) while ethyl acetate is still present in the system. The swelling ends when the cubic D surface which coexists in excess of the water-rich axis is obtained.

Interestingly, the cubic ice region, according to Figure 1, exists at relatively high EtAcinnel, see Figure 1. Another important feature is that EtAc is not completely miscible with water. This rate made it possible to combine a polymer-rich phase, consisting of PLG / MO / EtAc, in an aqueous state because the polymer-rich phase, which also contains some MO and EtAc, can be in thermodynamic equilibrium with a very aqueous rich mo. This is inferior to the level with which the NMP so strongly prefers water (forming NMP (water) 2 complexes) and the polymer polymer. Also note that EtAc, to some extent, is soluble in water. The maximum solubility is 8% EtAc in water at 20 degrees C. Through the varicose vein composition, the rate at which EtAc leaves the emulsion droplets / particles can be varied.

The lipid and polymer show a strong annual separation. By choosing a suitable solvent, the lipid / polymer interaction can be reduced so that a liquid phase, the so-called L-phase, exists. Genomic / lip-polymer composite particles are formed, according to the above-mentioned Characterization, of lipid / polymer composite particles, by confocal laser scanning microscopy, showing that distinct water-swellable lipid domains formed within the particles when the polymer particle concentration increased. Finally, the water-containing lipid domains leave the polymer matrix. Figure 3 schematically illustrates the lipids described in Example 1. Lipid / polymer composites have a particularly effective substance and for applications in the field of controlled. The quaternary system exhibits a suitable lipid / polymer composition. The particles can be formulated so that different, desirable, properties are obtained. Examples of such properties are; threshold size, structures, etc. The composite particles are suitable for implantable, depot and injectable drug systems for delayed fi insertion of therapeutically active substances. Furthermore, the particles may be useful in inhalation or oral delivery of active substances. Another interesting application is "site-specific targeting" video cancer therapy.

The lipid / polymer composite particles must contain at least one biodegradable polymer. The polymer may be a homopolymer, a random block copolymer or a mixture thereof. The polymer must be partially or completely soluble in organic solvents and not be completely soluble in water. A biodegradable material can be degraded to lower molecular weight units and eliminated by the living organism. A biodegradable polymer can, for example, be built from biodegradable nionic units. Examples of such copolymers (copolymers) that can be created from such monomers are: poly (actide), poly (glycolide), poly (p-dßmmn fl), p ° 1y (wx> r ° 1ß <= t ° n °), p ° ly (hydr °> = ya1kanøate), polz / (vwpylßnß fi lm fl ratß), poly (orthoesters), pol fl phosíiateesters) and poly (anhydrides). Combinations of these 500 505 510 515 520 525 530 535 540 545 527 505 homopolymers can also be used. Examples of copolymers are: various poly (<1,1-lactide-co-glycolide) polymers or other biodegradable, or biocompatible, copolymers.

The lipid / polymer composite particles should contain at least one lipid, or other surfactant molecule, with the potential to form desirable bilayer- or monolayer-containing fi axis (s) in water rich environments. The desired detiasene cantex, be (fair value / reverse) cubic, La, lamellar or hexagonal. Vesicular or micellar arrangements may also be of interest. Both synthetic and natural polar lipids (or other am fifi la molecules) can be used. The lipid may be anionic, cationic, or uncharged.

Appropriately used lipidate, the idea presented, is monocharged monoglyceride, called mononoolein or glyceryl monooleate, which is polar with the ability to swell, and which spontaneously forms a cubic bicontinuous phase in excess water. The Dcubian ice has a nerdynamically stable structure, built of lipid attachments that separate two separate water-channel systems. The only difference between two water channel systems present in a cubic phase is due to the weight of the excess water, which is the opposite of the water channel with the other water channel system being closed.

Examples of other lipids of interest, which may also form a cubic phase, are: monoelaidine, phosphosatidylethanolamine, phospholipids and PEGylated phospholipids.

Figures 4 and 5, the CLSM images present the particle characterization, which is further described in Example 2. The results show that lipid domains are present in the lipid / polymer composite particles but not in the reference particles. Dü fl ir the donors consist of MO (or M0 with water).

When MO comes into contact with water, the M0 domains begin to swell, ie incorporate water into the attachments. From the phase behavior of the EtAcMO / water system, it is noteworthy that the lipid, due to the water swelling, first forms a swollen liquid.

As a result of further swelling, the lipid undergoes transformations; L-Ld- k fl bi fl k (Qe), cubic fl k (Qin) - By varying the composition of the new kinase kinetics in the domain formation is controlled. In the case of 100% external water, M0 immediately separates from PEG-PLG (<10 minutes). When 4% EtAc is used on external fi ice, MO separates and forms domains after approximately 10 minutes. Since the outer Ibsen consists of 6% EtAc, M0 forms dormers after about 16 minutes (smaller particles). Ihlletmedenytue fi isrnâttadmedEtAqdvrß 'V>, fasseparerirttesystemetinom9O minutes.

In all experiments, the slow kinetics on the ice separation, ie the near-neutral phase, consisted of 6-8% EtAc, there are "large pore similarities" inside the emulsion droplets (lu or possibly EtAc domains or outer read). The pores, which are presented in Figure 4, are about 3-9 micrometers. When the solvent leaves the emulsion droplets, MO domains are then formed. Innessamärattlipiddonñnermbotjarsvä fl a fi ånrdgrasådanaporer / vâtske domäner. At the tkl point of swelling, porerm contains white ice. 527 505 ll 550 When the kinetics of phase separation are fast, ie when the outer fi axis of pure water, no pores have been observed. Water-swollen lipid domains are formed immediately. In any case, common to all fi tll with domain formation, regardless of the kinetics of the ice separation, is that the domains begin to swell before they alarm 555 p The particles are polydisperse (1-100 micrometers). M0 tends to form domains faster in smaller particles than in larger particles. In fact, the smaller the particles have a larger surface / volume retention and therefore the transport of EtAc (as well as the transport of water) becomes more efficient (ie shorter dilution path through the 560 polymer matrix and a relatively large surface area). The particles examined with CLSM year, due to practical reasons (immobilization of the particles) 15-35 micrometer diameter. However, particles 1-10 micrometers in size can be easily formed.

The size distribution is then clearly more monodisperse. Nanoparticles can also be formed within the framework of the invention. 565 Incorporation of therapeutically active substances into the lipid / water domain dom na) 570 Therapeutically active substances may be; (i) hydro fi la, (ii) hydrophobic or (iii) surfactants and interesting in various different areas, such as drug applications, cosmetics and functional foods, where the active substance exerts an effect on the living organism. Below are presented some interesting 575 In the case of hydrophilic therapeutically active substances, it is sufficient to dissolve the active component in a small volume of water (eg 100 microliters). The L-fitsen, consisting of EtAe / MO / (PEQ-PLG can for some also also dissolve some water. If this is the case, the small water phase (with active hydro fi l component) can be dissolved directly in the L-phase. Example 1.5 580 Water addition to the EtAe / MO / PLG system may, however, induce MO / PLG ice separation. If the lipid radical aliaser is present, the polymer-rich phase just before the particle formation occurs according to Example 1. 585 In the case of hydrophobic therapeutically active substances, these can be dissolved. The L-ice is formed and particle formation takes place according to Example 1.

In the case of surfactants, the active substance is suitably added to a lamellar (or cubic or an L; i fi ll with mixed solvents and / or 590 fi Hsatsermvânds fl asDvs fi HMOiäsenadderasHtevatæmen fi gt giventent fi gurentillagan to be active). The phase should be incubated, at the appropriate temperature and time, until the phase is macroscopically homogeneous EtAe / PLG is then added until a suitable L-phase is formed. Particle formation takes place, eg in accordance with Example 1. 595 Particle formation can also take place by other, within the tent de níred, methods (eg SPNY IOYKI fi IIQ, Wfud fl ríns, Spf fl y fl í fl sl 600 605 610 615 620 625 630 635 645 5245 formulations The solid, gel-like matrix By using a water-insoluble polymer the soluble formulations can be formed and by forming a slightly anhydrous polymer the latter can be formed.

The form of the final formulation (particles, surfaces, impl fl l flfl f, CIO) b ° f ° f W! * W the composition of the polymer-rich samt axis and kinetics of solvent removal. To list some parameters that control / affect the properties of the Shl fl iâ fl m fl ïe flfl let, Riljande can be noted; (i) the properties of the polymer such as; structure, molecular weight and glass transition tempexatm (ii) kinetics for solvent removal; ie choice of solvent and manufacturing method as well as composition and volume proportions An example of a formulation is particles, where the polymer is first dissolved in the lyase and then emulsion droplets are formed, according to Example 1, in an outer aqueous phase. Particles are formed as a consequence of the removal of solvents. The chemical system, which is more useful than the particulate matter, is further described below; Particles and (swelling) lipid domains are dispersed in a polymer matrix with particles having a lipid (or polymer) core placed inside a polymer (or lipid) shell and porous polymer particles, interesting to use for applications such as inhalation.

Furthermore, the chemical system can be used to form solid to semi-solid matrices, useful for, for example, depot Applications General Applications Generally, system-appropriate applications such as encapsulation of therapeutically active substances (eg proteins, peptides, vaccines, DNA, RNA).

M0 has mncoadhesive properties which means that lipid / polymer composite particles are useful for oral delivery. However, for protein applications in general and in particular the delivery of other therapeutically active substances - which must act in a specific place - so-called "site-specific target", other routes of administration may be interesting.

The invention is interesting to use in the manufacture of seedlings in granular iberta tableting.

The lipid (or amnn am fifi l compound) then acts as a binder in the final formulation. The present invention offers a way to make 650 655 665 670 675 680 685 690 695 527 505 IH granules from a "one-step process" provided that the desired strength for the phase separation, between the polymer and the binder (lipid or other compound), is used.

Examples of specific active substances and formulations The applications listed below are suggestions for possible applications. The majority of other applications which may also be of interest in the present invention present the invention.

As an example, the encapsulation of calcitonin in the filtration can be used, which can, for example, be designed as an injectable or implementable depot effect implant. The problem with calcitonin, as with other peptides, is that it retains its activity during encapsulation. The system, Y fi PPOIICIat iupp fi nníngen, is interesting in encapsulation of, for example, calcitonin.

Delivery of AVNA is one specific example of how it can be used.

As DNA has been reported to pack well in lamellar axes, the use of cationic molecules appears to coexist - subbalance (thermodymic or kinetic) co-water entrails, product-specific concepts for DNA encapsulation and encapsulation.

It has presented the invention, if it involves the formation of domains of, for example, cationic amyl molecules (with DNA) located in connection with a biodegradable polymeric matrix (which in this case may be charged). The polymer can be formed, for example, as follows; dispersed domains within a matrix of the amiolacyl / contaminated matrix / core or shell particles. The phonmylations, such as particles or matrices, are used in the field of molecular applications or applied in molecular biology (eg transfectins or pure method applications). Another specific example is depot-type formulations. Nicotine has special solubility properties, which means that encapsulation of nicotine in a coniposit material can be extremely interesting, since nicotine is then distributed in both hydro- and hydro-oba domains, which then separate in vivo. (Nicotine treatments can consist of a concentration peak followed by a ird-delayed itt sitting or as the smoker, o fi a wanting a lazy kick). Interesting formulation types can be, for example, shell / comb particles and matrices.

Further examples are insulin-based formulations, such as pulsed and / or controlled insertion of insulin. An interesting application is to encapsulate PEG-conjugated nicotine within PEGylated composite lipid / polymer material.

Porous particles, described below, can be used as a means of inhalation. If the composite material is designed into a porous implant, this may be of interest for applications in, for example, tissue regeneration and so-called regeneration and scaling - where living cells and (essential) growth factors may be suitable for the benefit of the variability of the porosity of the material. 527 505 lb Further examples are combining smart polymers to form smart composite 700 materials in accordance with the invention presented herein.

Notably, the composite materials include, but are not limited to, particles or implants. The deposable implants km, for example »are produced according to a two-step process: 705 First, the L-fiase, containing polymer (PLG) / am am l molecule (M0) / solvent, is formed. The active substances are added as described above "Incorporation of therapeutically active substances into lipid / water domains (s)". A phase region of the ternary PLG / MO / EtAc system, presented in ñgnr 1, de fi níers possible of the L- fi sen. It is possible to: vary the concentration of MO and PLG in a wide range, ie the MO content can be varied from 0-90% by weight and the PLG content can be approximately varied between 0-100%. In a second step, the implant is titrated by casting or extruding the L-tube under simultaneous contact with external water. Alternatively, the casting / extrusion takes place during simultaneous heating during which the volatile solvent evaporates. 715 Release of active substance These are the formulation arms that present interesting applications such as cannulated or delayed release of active substance. 720 of encapsulated active substance may generally, regardless of the type of precursor, for example depend on; (i) hydrophilicity / hydrophobicity of active substance. The active substance is localized to the radiolicadonians of the crucible, depending on the molecular structure of the active component. Hydro active substances will mostly be located in the aquifer of the cubic (or amian phase consisting of monolayers, or bilayers) ice.

Hydmibba substances will mostly be located in the bi-layered bi-layered bi-layer (or other mono- or annex 730-containing phase). Surfactants will preferably be located on the interface between the hydrophobic (here exemplified by MO) and the hydro (eg water) domain. (ii) The definition of the active component depends on the molecular weight. (iii) Hydrolysis, and water uptake, depends on some specific properties of the 735 polymer, such as the lactide / glycolide ratio, molecular weight, degree of PEGylation in all when a PEGylated polymer is used, as well as temperature and pH.

Notably, the nature of the active substance will depend on the 740 mechanism with which the active substance is inserted. The mechanism is dependent on the type of formulation. Finally, in addition to the formulation type, it also depends on, fi iljande; active component, polymer properties, and type of phase (rnono / appendices containing ice), etc. 745 Formation and characterization of the lipid / polymer hybrid particles The method of pair formation, which is a part of the invention, is described here: 750 755 760 765 770 775 780 785 790 795 527 505 lb The principle behind the particle formation method, according to the invention, is to transport the solvent away, in a controlled manner, from a mixture or solution of a specific specific original mixture. The production method comprises the phase behavior of a block system whose phase behavior is presented in Figure 1.

The parent mixture can be either a single phase (L) or an emulsion, (as described above "incorporation of therapeutically active substances into lipid / water domains (s)"). The parent mixture consists of at least one solvent / at least one polymer and at least one lipid (or other am fifi l molecule). It is fitting that the source mixture is a true, thermodynamically stable, read, which means that fi then has great potential to preserve the stability during incorporation of active substance - which also means that the activity of active substance tends to be maintained (even if a lock transition is induced).

The removal of solvent can take place by a process which in some cases (ie assumes a relatively low solubility of the polymer solvent and the other solvent) can be likened in the solvent extraction to a second solvent.

The polymer is only slightly soluble and preferably insoluble in the other solvent, such as hot water. Thus, the polymer was contacted with the "second release agent".

Ermilsions fl lriarandet, described in example 1, is an example of this process.

Very interesting, in addition to the emulsion method, is the use of a spray drying process in transporting away the properties of the polymer.

The initial solution is then sprayed and the polymer solvent evaporates, while simultaneously concentrating the polymer phase, which means that solid to semi-solid particles can be formed, eg depending on the choice of polymer, molecule, and solvent.

The polymer must have the following properties; the polymer should not be fi intimately miscible with water ('the second solvent'). Preferably the polymer should be insoluble in the second solvent. At the very least, it should be convenient to concentrate the polymer, by elimination, in the presence of the second solvent.

EXAMPLES Example 1: Manufacture of lipid (am flfl D / polymer composite particles Lipid / polymer granule positive particles, presented in Figure 4, with domains dispersed in a polymer matrix, can be formed according to the following procedure where a liquid (L-phase) is emulsified in an aqueous kingdom.

The following procedure is used for the formation of the L-phase: The lipid probe, Lissamine m rhodamine B LZ-dihexadecarioyl-sn-glyeeroß-phosphoethanolamine triethylammonitime salt (rhodamine DHPE), is first dissolved in at least 95% ethanol and then mixed with M0. The lipid / probe solution is incubated at 50-70 degrees C until the ethanol has evaporated. Polymer and solvent are then added so that only a single solution, L-iiisen (see Figure 1), is obtained.

The L phase used to form reference particles presented in Figure 5 contains the same amount of DHPE probe, PLG and EtAc as the L phase used to form lipid / polymer composite particles. The only difference is that the latter also contains some M0. 800 805 810 815 820 830 835 845 527 505 ll Particles are formed by emulating 0.1 ml of L phase (the composition of the L-phase is given in Example 1) in the KinematicaAG, PT3OO) somarbetariii minutervid4000rpm.

Example 2: Particle characterization THE PARICLES WERE CHARACTERIZED WITH mm By A 'xoNEoKAL IASER SCANNING Mncaosxor, CLSM. EMUwoNEN / dispersion was placed 1 a chamber with Err 17nMnocKrG1AsDo1TEmENn1AuoG1AscYLmDmMEDEN1NRED1AMEmaAv 15 mm, 1.1MMADEsPAcs1Asao'nEN.IrzNAN U¶róRDEsMEDCLSM (Leica TCS 4D, Leica LT, HEmEIBERG, GERMANY) mmomusEaADEs PARmuARNA THIS 'liLLsAïs of 0.2 mm glass KUwR. A '63x / 1.2 OBIEKTN IS USED AND 568 NM NANDERS OF AxGoN / I-: RYPION LASENN MUNAMNADE RHODAMINE "SCANNINGEN" mnonDEs WITH 8 LINNANS MEDEWARDE ocn UPPmsNmcmN WAS XYZ (now 12M / 0.12 pM / 0.12 pM 0.12 pM 0.12 pM. BHDERNA, WHICH SHOWS THE EWSSION OF THE NHODAMINE-DIIPE PROAEN (MOUSE) AND IN SOME CASES ALSO REFLECTION FROM THE LASER (MORK), AEARBERADAS AND UWÅRDERADBS WITH SOFTWARE IMAGE SPACLAVA AVENA THE WORK STANDARD USED BY SMOON GRAPmcs (MOUNTAm Vmw, CA).

LIPID phase Rated impure aHoDAi al NE DEIPE, ANvÅNDBs 1 PART Fóksóx sAMnDIGT WITH van phase Rated impure nomw DISULEONAIE the Fónsr said EmEnADEs of argon / KRYPION IASERNS 568 nm band ocHDEN sENAaE 488 nm BAND PAnmaAn WITH KAnNlsxAL smuicnm It ärocksåniöjligtamgenomattvariera lipid / polymer ßrßllandet , create a core / shell structure where the lipid (or polymer) forms a core with a polymer (or lipid) shell. As an alternative to the emulsion method, the spray drying method is interesting here (provided that a strong separation is used and a polymer with a suitable glass transition type is selected). . In such a formulation, the lipid content may be varied so as to obtain desired properties in conjunction with the formulation described in Example 1.

Porcelain polymer particles It is important to consider the chemical system according to the invention, for the production of highly porous polymer particles. These particles may be of interest inhalation as these, due to their porous structure, have the same aerodynamic properties as the smaller particles.

The perous particles can be formed according to the following two-step process; (i) Article formation and (un) washing. In the first step, lipid / polymer composite particles are created, for example, according to the detailed description given in Example 1. Another alternative is to form the particles by spray drying (after which the particles can be incubated in water). In the second 527 505 I? step, the water-swollen lipid domains are washed out of the polymer matrix, using a solvent which acts as a solvent for the lipid but not for the polymer. The final degree of porosity depends on the amount of lipid present.

It should be possible to create particles with a degree of porosity of approximately 5-70%. The deformed polymer particles should have a porous inner structure. Such particles, after addition of active component, may be of interest for applications within 855. The advantage of these particles is that a variability of the porosity can be obtained by varying the storage time in water, true kinetics during particle formation. 860 865 870 875 880 885 890 895

Claims (24)

905 910 915 920 925 930 935 945 527 505 Iq Krav A non-fibrous composite composite material, wherein the material included in the invention consists of at least one component component and the polymer component, wherein the method comprises the following steps: homopolymer, a (random) block copolymer or a mixture thereof (ii) denam fifi la ren lreningenlmr fi irn depending on the properties of the final material, it must have a retained or semi-solid material. The method of claim 1, wherein the step of wiping off the solvent consists in extracting the solvent against a liquid solution consisting of at least one second 'solvent'. 3. A method which relies on a high solubility agent is inadvertently miscible with such other solvents. 4. The method, referred to as the third solvent solvent component or mixtures of, for example, water, lower eyeloalkam, eg eyclohexane. The method of any one of claims 2-4, wherein the lipid / polymer mixture is an emulsion, and the emulsion is added to an outer aqueous phase, where particles are formed as a result of removal of solvent. The method of claim I, wherein the step of transporting the solvent consists in a spraying method in which the solvent is transported away. 7. The method does not require a specific clause and the other composite material material; particles, solid implants, semi-fi ista implants, gel-like matrices useful fi Sr applications '' in, for example; coating ', cosmetics, veterinary' media 'in, functional food, delivery of active components with controlled or id iidrójd The method as claimed in any one of the preceding claims, wherein the mono- or annex-containing isase; fi ist fl crystalline) state or arranged cubic, Lgeller La, lamellar or hexagonal (right-handed / inverted). 950 955 960 965 970 975 980 985 995 527 505 10 9. The method, referred to in any of the foregoing seas, where the total purification is selected from the groups of synthetic or natural polar lipids or other surface substances (so-called solids). 10. The method referred to in any of the preceding claims, wherein the whole component is anionic, cationic or uncharged. 11. The method, which is invoked in any of the above seas, where the am fifi la ren l purification is selected from compounds that have the ability to form, for example, a cubic phase. 12. The method, referred to in any of the above seas, where the am fifi la fi Purification is an uncharged monoglyceride, iöredrags glyceryl monooleate. 13. The method referred to in any of the above oceans, wherein the whole compound can be selected fi other than monoelaidine, phosphatidyl etharmlamine, phospholipids (eg lecrtm) and PEGylated phospholipids. 14. The method referred to in any of the above seas, where the polymer is partially or fi completely soluble in organic solvents but not completely soluble in the 'other solvent' (eg water). The method referred to in any of the above seas, wherein the polymer is a homopolymer selected from poly (laetide), poly (glyoolide), poly (p-dioxanone), p fl lnwpr fl laßf fl nß). wlnhywxyßlk fl w fl tß). wlyßvfovylenß (lm fl rate). polyorthoesters, polyphoshate esters, and polyanhydrides, and combinations of these hoxnopolymers. The method referred to in any of the above seas, where the polymer is a 'copolymer' selected fi than various poly (d, l-lactide-co-glyoolide) polymers or other biodegradable (biodegradable) or biocompatible copolymers or bioactive copolymers. 17. The method referred to in any of the above seas, where the fl solvent is only partially miscible or completely immiscible with water. Use of materials obtained by the method referred to in any of the preceding claims, in implantable, disposable, or injectable delivery systems fi ßr fllrrated / controlled by active component. 19. Use of particles obtained fi- by the method referred to in any of the provisions 1-14, fl in the manufacture of a formulation for inhalation or oral delivery of therapeutically active fi-formulation or substance. Composite materials, manufactured in accordance with claim I, consisting of a polymer matrix containing at least one domain of fl surface (crystalline) fi axis. 21. Materials referred to in ocean 20, with (crystalline) means of myelular or vesicular deworming, containing, for example, a water-rich, located (in cavities) within a continuous matrix. 1000 1005 1010 1015 1020 1025 1030 1035 1040 1045 527 505 ll Material, as claimed in claim 20 or 21, in the form of (nano / micro) particles. Materials as claimed in claims 20 or 21, in the form of implants, implants or depot systems, gel-like hydrogels, and for described and exemplary applications (coating, functional food, veterinary medicine, cosmetics, pharmaceutical technology). 24. A system, fl is attenuated or delayed fi insertion, consisting of nnterhl invoked in any of the seas 20-23).