Apparatus and method for producing nanocarriers and / or nanoformulations

Abstract

The present invention relates to an apparatus (0) and method for producing nanocarriers and / or nanocompounds, as well as the corresponding process product. The object of the present invention is to identify an alternative method for producing nanocarriers / nanocompounds and the associated apparatus for carrying out the method. The apparatus (0) according to the invention is characterized by the vertical orientation of the supply conduits (4, 5) leading to the active element (3). The supply conduits (4, 5) are nested within each other and axially movable with respect to their orientation. The method according to the invention mixes at least two liquid phases with different acidities. The volumetric flow rate of the first phase is greater than the volumetric flow rate of the second phase.

Description

[Technical Field]

[0001] In the context of the present invention, a nanocarrier is a composition for carrying an active ingredient of a pharmaceutical, cosmetic, or dietary supplement. A nanocarrier may consist of a pure substance or a mixture of two or more substances. The substance may be solid, semi-fluid, or liquid. The substance may be a single-phase or multi-phase substance, and in all cases, the nanocarrier is in the form of particles, with an average particle size of less than 300 nm.

[0002] Examples of nanocarriers include particles of natural or synthetic polymers, lipids (lipid nanoparticles - LNP), liposomes and micelles, and nanoemulsions.

[0003] Nanocarriers composed of two or more substances are generally prepared by preparing the individual substances or their precursors as dispersions or solutions in a liquid medium and then mixing them together. Mixing results in physical interactions between the individual substances or precursors, forming nanocarriers. The nanocarriers may optionally be subjected to subsequent post-processing, such as separation from the liquid medium. The resulting nanocarriers may then be used as placebos or loaded with active ingredients to form nanoformulations.

[0004] In the context of the present invention, a nanoformulation is a dosage form of a pharmaceutical, cosmetic, or nutraceutical active ingredient loaded onto nanocarriers, which may be on the surface of the nanocarriers, inside the nanocarriers, or complexed to the nanocarriers.

[0005] Examples of nanoformulations include, inter alia, so-called lipoplexes or polyplexes, i.e. complexes of polymers or lipids with, for example, DNA, RNA, proteins, peptides, etc. Within these complexes, the polymer / lipid forms the nanocarrier, while the DNA / RNA forms the active ingredient.

[0006] As mentioned above, nanoformulations can be prepared by loading active ingredients onto nanocarriers.

[0007] However, in pharmaceutical technology, it is beneficial for the economics of the process to carry out the production of nanocarriers and the loading of active ingredients onto them in an integrated manner: In such integrated methods, nanocarriers are first prepared and then loaded with a nascent active ingredient. This is typically accomplished by preparing the individual components of the nanoformulation as a dispersion or solution in a liquid medium and mixing them. These components include the nanocarriers / precursors of these materials and the active ingredient or its precursor. Mixing results in physical interactions between the individual components, forming the nanoformulation. If necessary, the nanoformulation can then be subjected to further post-processing, such as separation from the liquid medium.

[0008] In particular, nanoformulations using LNPs or polyplexes as carriers are always produced in an integrated manner, where the loading of the active ingredient onto the carrier is carried out in a nascent state.

[0009] A crucial aspect of equipment engineering in nanocarrier / nanoformulation manufacturing is the design of the mixer that mixes the components dispersed or dissolved in the liquid medium.

[0010] In conventional mixers, mixing itself is a challenge. Therefore, mixers are miniaturized into so-called micromixers. In such micromixers, a main stream is split into many small substreams, which are then mixed together. These small structures are very sensitive to fouling (deposits on the walls) and small air bubbles. Therefore, reactants for micromixers must be prepared to meet high purity requirements. This means, in particular, that dissolved air or gases such as nitrogen must be painstakingly removed from the liquid.

[0011] EP 1519714 discloses a method for producing nanocompounds using a device with a T-shaped mixer. The two fluids to be mixed pass through coaxial supply conduits to a collision point, where they are mixed and then withdrawn through outlets offset by 90°. The angle between the two supply conduits is 180°. The volumetric flow rates of the two fluids entering the active element are the same.

[0012] EP 1937213 discloses a method and apparatus for producing nanoformulations, using two T-shaped mixers arranged in series. Thus, three fluids can be mixed in two stages. A drawback of this configuration is that the flow conditions created by the T-shaped mixers change with increasing production scale (the so-called scale-up problem) (hydrodynamic similarity problem). Changes in flow conditions result in changes in product characteristics. Therefore, adapting operating conditions and pilot runs optimized for product quality in the laboratory to larger production scales is not trivial. Instead, industrial operations involve simultaneously operating multiple small-scale devices under conditions optimized on a pilot scale ("numbering up"). This increases capital costs. A further drawback of this method is that nanocarriers are not stable for long in the environment of the mixture formed in the production of LNPs. Therefore, nanocarriers must be separated from the mixture particularly quickly or stabilized in an additional step. Both options significantly complicate process management.

[0013] EP 3711749 A1 discloses a Y-shaped mixer for producing nanocompounds. Because of the Y-shape, the two feed lines to the impingement point are not parallel. A Y-shaped mixer can also be combined with a T-shaped mixer to produce a three-phase mixture. The Y-shaped mixer also has complex flow dynamics, and therefore, only by "numbering up" can it be moved from pilot scale to production scale. The problem of insufficient stability of LNPs in the resulting mixture also arises in Y-shaped mixer processes.

[0014] WO 2001 / 005373 discloses a mixing head for producing nanoformulations, which has a plurality of injectors with a diameter of less than 2 mm.

[0015] WO 2017 / 223135 discloses a mixing head that also operates according to the injector principle, the injector being provided with a metering device in the form of a servomotor-operated pipette.

[0016] The objective of the present invention is to identify an apparatus for producing nanocarriers / nanoformulations in which the hydrodynamic ratio is maintained during scale-up, thereby allowing for increased production scale without "numbering up".

[0017] Furthermore, it is believed that the method for producing nanocarriers / nanoformulations carried out using this device will reduce the number of operational steps.

[0018] This problem is solved by a device according to claim 1 and a method according to claim 13 that can be carried out using this device.

[0019] The present invention therefore: a first storage vessel for containing a first liquid phase; a second storage vessel for containing a second liquid phase; an active element for mixing a first liquid phase with a second liquid phase to provide an at least two-phase mixture; · a first supply line fluidly connecting the first reservoir to the active element; · a second supply line fluidly connecting the second reservoir to the active element; · A collection container to contain the mixture; · a discharge passage that fluidly connects the active element with a collection vessel; a pump integrated into the discharge channel so that the mixture can be conveyed by the pump from the active element through the discharge channel into a collection container; The present invention provides an apparatus for producing nanocarriers and / or nanoformulations, the apparatus having the following configuration:

[0020] According to the invention, the device comprises: the first supply channel and the second supply channel are vertically oriented at least in a vertical section; In the vertical section, the second supply channel is arranged inside the first supply channel; and In the vertical section, the first supply passage, or the second supply passage, or both supply passages are axially movable such that the axial position of the second supply passage relative to the first supply passage is adjustable; It has the following configuration.

[0021] The device according to the present invention is characterized by the vertical orientation of the feed channels leading to the active elements: both the first and second feed channels are vertically oriented in at least one section upstream of the active elements, so that the flow through the feed channels is directed in the direction of gravity acceleration. In this specification, the vertical section of the feed channel is referred to as the vertical section. This is not a specific structural element of the device, but rather the area of ​​the device in which the feed channels extend vertically. Since the flow direction through the vertical section is parallel to gravity acceleration, i.e., downward, and the active elements are located downstream of the feed channels, the vertical section theoretically extends above the active elements. However, the feed channels may be horizontal or inclined outside the vertical section, so the vertical section does not necessarily have to be vertically positioned above the active elements. Nevertheless, it is preferred that the vertical section be vertically positioned above the active elements, and it is particularly preferred that the vertical section extend up to the active elements, so that the liquid phase enters the active elements vertically. In this configuration, the flow vector is oriented parallel to the gravity vector.

[0022] A further essential feature of the device according to the invention is that the second supply channel is led into the first supply channel in the vertical section. This means that the second supply channel is surrounded by the first supply channel. The walls of the second supply channel are therefore in contact with the second liquid phase on their inside and with the first liquid phase on their outside. The walls of the first supply channel are in contact with the first liquid phase on their inside and with the ambient environment on their outside.

[0023] The vertical orientation of the feed channels and their nested arrangement have a favorable effect on the flow conditions in the active elements, resulting in a particularly homogeneous mixing of the phases. Furthermore, the flow ratios provided by this configuration have proven to be particularly stable when the flow cross section is changed. Thus, even operating conditions optimized for small flow cross sections are maintained during scale-up.

[0024] In principle, the active elements of the device form a thin film for the mixture transport. The ratio of capillary flow to edge flow of the buffer medium is relevant. The thin film formed (telescope antenna effect) allows mixing essentially by diffusion, without the occurrence of deposits or blocking at the interface. Since the device according to the invention does not require "throttling" the flow through the microstructure, the procedure proceeds practically without pressure, which allows for easy scaling up of the arrangement and the method.

[0025] A further design element of the invention is that the first and / or second feed channel is axially movable in the vertical section, so that the axial position of the two feed channels relative to each other can be adjusted. By varying the axial positioning, the flow conditions within the active element can be adjusted, thereby optimizing the mixing results.

[0026] The pump of the device is preferably a centrifugal pump comprising a housing and a rotor (impeller) rotatably mounted within the housing about its axis of rotation and rotatably movable via a propulsion means. The simplest example of a propulsion means is a motor mechanically coupled to the pump rotor via a shaft. Suitable alternatives include a magnetic transmission or a mechanical shaft. In either case, the rotor's axis of rotation is preferably vertical.

[0027] The vertical rotation axis of a centrifugal pump means that it accelerates the mixture in a horizontal plane. The fact that the liquid phase flows downward through the vertical section and then accelerates horizontally after mixing means that the liquid makes a 90-degree turn in the flow path through the feed, active element, and pump. This enhances mixing. Thus, the centrifugal pump also plays a part in the mixing process, which is effectively a second mixing stage. A vertical rotation axis can also slightly destabilize the rotor, which allows the pump to be used for quenching. Furthermore, if the rotation axis and the axis of the vertical section are oriented coaxially, the mixture can sometimes be introduced into the housing area and transported by wall flow. This also has a positive effect on product formation.

[0028] All these design elements according to the invention result in intensive mixing of the liquid phase in particular, so that the components present therein can come into particularly intimate contact with one another and interact efficiently. The powerful diffusion processes resulting from the large difference in speed ensure a fine distribution of the nanoscale materials in the dispersion medium. As a result, mixtures of nanocarriers / nanocompounds with a particularly narrow particle size distribution can be obtained.

[0029] A further advantage of the device according to the invention is its simple and robust construction, which increases its reliability. Furthermore, the device can be operated both batchwise and continuously. Therefore, in the same device, the production process can be optimized first in batch operation and then easily switched to continuous operation to increase the production capacity. Furthermore, the flow conditions within the active elements have proven particularly stable, so that increasing the diameter during the capacity increase process only produces minimal analogous effects that could impair product quality. The device according to the invention thus allows for practical scale-up for increasing the production capacity. This allows for particularly economical transfer of knowledge from experimental operation to industrial production-scale operation.

[0030] Ultimately, the device according to the invention allows for the development of cost-effective processes and consistently high quality production, thus making the invention particularly economical to produce.

[0031] A further advantage of the device according to the invention is that it is less susceptible to fouling and blockages, since its active elements repel microstructured components.

[0032] According to a preferred development of the device, the first supply line in the vertical section is formed from a first linear pipe, and the second supply line in the vertical section is formed from a second linear pipe, with the first and second linear pipes extending coaxially along the vertical section. In this embodiment, the supply conduit has the form of a fixed pipe and extends coaxially at least along the vertical section. The rigid pipe allows for particularly stable flow conditions and precise axial positioning. Outside the vertical section, the supply conduit may be flexible, for example in the form of a hose line. This may also be necessary for axial mobility. The coaxiality, i.e., the central placement of the second supply conduit within the first supply conduit, allows for a particularly stable flow profile in the active element.

[0033] It is preferred if the second linear conduit is in the form of a capillary tube with a cross section significantly smaller than that of the first linear conduit, since, according to the invention, the flow (volume flow rate) through the first supply conduit is much greater than the amount of second liquid phase added. Preferably, both the first and second linear conduits have a circular cross section. It is also possible for the second supply conduit to be doubled, i.e., to consist of two parallel conduits or capillaries, each of which carries half the volume flow of the second liquid phase.

[0034] In its simplest configuration, the device according to the invention is only capable of producing a biphasic mixture of a first and a second liquid phase (a binary system), however, several variants are possible, such that the device can produce at least a triphasic mixture of a first, second and third liquid phase (a ternary system).

[0035] In this specification, embodiments of the apparatus capable of producing only biphasic mixtures are referred to as "binary mixing apparatuses," and embodiments of the apparatus capable of producing triphasic mixtures are referred to as "ternary mixing apparatuses."

[0036] All variations of the three-way mixing device must have a third storage vessel for containing a third liquid phase and a third supply passage fluidly connecting the third storage vessel to an active element configured to provide at least a three-phase mixture by mixing a first liquid phase with a second liquid phase and a third liquid phase, the first supply passage in the vertical section being formed from a first linear conduit, the second supply passage in the vertical section being formed from a second linear conduit, and the third supply passage in the vertical section being formed from a third linear conduit. According to the general concept of the present invention, the third supply passage is vertically oriented at least in the vertical section, is disposed inside the first supply passage in the vertical section, and is axially movable in the vertical section such that the axial position of the third supply passage relative to the first supply passage is adjustable. The third supply channel therefore meets the same design characteristics as the second supply channel.

[0037] In a first variant of the triple-mixer, the second and third lines run parallel to each other through the first line. In the simplest case, this means that the walls of the third line are in contact with the third liquid phase on the inside and the first liquid phase on the outside. (The walls of the second feed line are also in contact with the second liquid phase on the inside and the first liquid phase on the outside.) The second and third lines are preferably configured as capillaries with a smaller cross section than the outer line. In this case, the lines cannot be oriented coaxially. The configuration of this triple-mixer is essentially the same as that of a double-mixer with a second line.

[0038] In a second preferred variant of the three-way mixing device, the second and third conduits may extend parallel to each other through the first conduit. The three conduits are not coaxial. The first conduit may have an annular cross section, which is achieved by providing a separate inner wall in the first conduit. The first liquid phase then flows through an annular gap formed between the inner and outer walls. The first conduit thus has a core through which the first liquid phase does not flow. Meanwhile, the second and third conduits pass through the core. The core remains empty between the outer walls of the second and third conduits and the inner wall of the first conduit.

[0039] However, in a third variant of the three-way mixing device, all three pipes may extend coaxially in the vertical section. This is possible if, in the vertical section, the third supply line is disposed within the second supply line, while the second supply line extends within the first supply line. Thus, the third pipe is surrounded by the first and second pipes, while the second pipe is surrounded only by the first pipe. The walls of the third pipe are in contact with the third liquid phase on the inside and the second liquid phase on the outside. The walls of the second pipe are in contact with the second liquid phase on the inside and the first liquid phase on the outside. The walls of the first pipe are in contact with the first liquid phase on the inside and the surrounding environment on the outside. In this three-level nested arrangement, all three pipes may, but need not, extend coaxially. However, a coaxial arrangement is preferred for stable mixtures. This three-stage nested variation is somewhat more complex than the first variation with parallel second and third lines, but provides better homogeneity during mixing.

[0040] In a preferred embodiment of the invention, the rotor's rotation axis is aligned coaxially with the axis of the first feed channel in the vertical section. This means that the pump, or more precisely its rotor, must be positioned below the active element. The mixture leaving the active element passes directly over the rotor, where it is accelerated horizontally outward. The pump rotor functions as an additional mixer, providing a finer distribution of the components in the dispersion. In this configuration, the pump discharge is preferably at least partially horizontal. Therefore, the feed and discharge channels are perpendicular. The flow is bent 90° within the pump. This configuration allows for the rotor to be destabilized, resulting in less recirculation, which has a beneficial effect on the product.

[0041] In a further preferred embodiment of the device, the pump is a magnetically levitated bearingless motor (Livitronic) pump. This should be understood to mean that the rotor is magnetically mounted and the propulsion means is a rotating magnetic field. The rotating magnetic field rotates the pump rotor, thereby enabling power transmission from the propulsion means to the rotor without mechanical contact between the propulsion means and the rotor. In this arrangement, the pump rotor is simultaneously the rotor of the electric motor, and the pump housing is simultaneously the stator of the electric motor. The pump and the propulsion means are integrated. The advantage of this configuration is that there is no drive shaft, so the housing does not need to be perforated by a drive shaft. In Livitronic pumps, instead of mechanical power transmission via a drive shaft, power transmission is purely magnetic via a rotating magnetic field. The rotating magnetic field may also affect the mounting of the rotor within the housing. This not only eliminates the need for a fragile seal on the drive shaft, but also allows the mixture to flow on both sides of the rotor's rotation plane, since there is no drive shaft to block flow on both sides. Another benefit of the Livitronic centrifugal pump is that the pump operates to minimize cavitation effects, resulting in less wear or spalling of materials that can contaminate the mixture.

[0042] Levitronic pumps are commercially available, for example as PuraLev-i30 type pumps, from Levitronix GmbH (Zurich, Switzerland).

[0043] One advantage of the apparatus according to the present invention is its ease of conversion from batch to continuous operation. All that is required is to recirculate the discharged product to the first storage vessel, form a circuit (loop) from the first storage vessel through the first supply line via a pump to the active element, and return to the first storage vessel. The collection vessel and the first storage vessel are identical, and the discharge line has the form of a circuit. This constitutes a particularly preferred embodiment of the present invention. In this arrangement, the product is collected from the first storage vessel after a given operating time of the apparatus or discharged from the circuit. The circuit does not necessarily have to be closed: it is also possible to circulate a relatively large liquid volume through the circuit of the apparatus while simultaneously discharging a relatively small amount of product from the discharge line. The storage vessel is then simultaneously replenished with new liquid phase according to the collected amount. This embodiment of the method is a hybrid of continuous and batch operation.

[0044] The volumetric flow rate of the first liquid phase is generally significantly greater than that of the second and / or third liquid phases. Therefore, to control the volumetric flow of the first liquid phase, it is sufficient to adjust the volumetric flow rate of the pump. The volumes of the first and / or second liquid phases can then be neglected. Therefore, in the simplest case, the device has a pump whose delivery volume can be adjusted. The delivery volume of the pump should be controlled in an open or closed loop. The delivery volume is most easily adjusted by the driving speed, i.e., the speed of the rotating magnetic field that rotates the rotor of the pump / motor.

[0045] To accurately adjust the mixing ratio, the apparatus should include at least one metering device configured to meter the second liquid phase into the active element. Since the volumetric flow of the second liquid phase is significantly smaller than that of the first liquid phase, in a preferred variant, the metering device can be a linear piston pump. The capacity of a piston pump is generally limited by the piston volume. While this is not particularly problematic in batch operation, it is a problem in continuous operation. Therefore, in continuous operation, multiple piston pumps are required, each alternating between metering the same phase. While one piston is metering, the other piston is lifted. A similar effect can be achieved by using a piston pump with multiple pistons.

[0046] The three-way device should also include a corresponding metering device for metering the third liquid phase into the active element. The same applies as specified above in relation to the metering device for the second liquid phase.

[0047] Preferably, the delivery flow of the metering devices, in particular of all metering devices, is adjustable. In the case of piston pumps, the volume flow is adjusted by adjusting the linear drive speed.

[0048] It is particularly preferred that the volumetric flow of the centrifugal pump and the metering device is controlled by a central control unit of the device.

[0049] The present invention also provides a method for producing nanocarriers and / or nanoformulations, which is carried out using the devices described herein. The method according to the present invention comprises: - providing a device according to the invention; - providing a first liquid phase containing a first liquid dispersion medium in a first storage vessel; - providing a second liquid phase into a second storage container, the second liquid phase being a second liquid dispersion medium and containing at least one component selected from the group consisting of a precursor of the nanocarrier, a precursor of the active ingredient, and the active ingredient; - operating the pump to establish a liquid flow from the first storage container through the first supply line into the active element and through the discharge line into the collection container; - metering a second liquid phase into the active element via a second supply line, the volumetric flow rate of the second liquid phase in the second supply line being less than the volumetric flow rate of the liquid flow in the first supply line; - mixing the first and second liquid phases in the active element to obtain a mixture containing nanocarriers and / or nanoformulations; - collecting the mixture in a collection vessel; - withdrawing the mixture from the apparatus; Includes.

[0050] The mixture is optionally further post-processed in a further step, which may in particular involve sterile filtration or separation of the nanocarriers and / or nanoformulations from the mixture.

[0051] This process is carried out in one go until a sufficient amount of nanocarriers / nanoformulations is accumulated in the collection vessel. The mixture is then extracted from the device. If necessary, the mixture is subjected to further post-processing, which in particular involves extracting the actual target product (nanocarriers / nanoformulations) from the mixture. Separation is carried out by evaporation of the liquid dispersion medium, or by filtration, for example, tangential flow filtration, or membrane technology, or by a combination thereof, according to conventional techniques. Further suitable separation methods include dialysis, sterile filtration, spray drying, or freeze-drying.

[0052] The triple blending device can be used to produce ternary blends, thus making a wider range of products available. The associated production method is: - providing a first liquid phase in a first storage vessel, the first liquid phase comprising a first liquid dispersion medium, the first liquid phase having a pH in the range of 6 to 8, preferably 7; - providing a second liquid phase in a second storage container, the second liquid phase comprising a second liquid dispersion medium and at least one precursor of the nanocarriers and / or the active ingredient, the second liquid phase having a pH in particular between 3 and 5, preferably 4; - providing a third liquid phase into a third storage container, the third liquid phase comprising a third liquid dispersion medium and at least one further component, the further component being selected from the group consisting of a precursor of the nanocarrier, an active ingredient, and a precursor of the active ingredient; - operating the pump to establish a liquid flow from the first storage container through the first supply line into the active element and through the discharge line into the collection container; - mixing the first, second and third liquid phases by metering the third liquid phase into the active element via a third supply path, wherein the volumetric flow rate of the third liquid phase in the third supply path is less than the volumetric flow rate of the liquid flow in the first supply path.

[0053] The production of three-component systems is of particular interest in the production of LNPs.

[0054] An essential aspect of the method for preparing a ternary mixture is that the first and second liquid phases have different acidities. The first liquid phase is closer to neutral and has a stronger buffering capacity than the second liquid phase, with a pH of 6-8, preferably 7. The second liquid phase, on the other hand, is more acidic, with a pH of 3-5, preferably 4. Because the volumetric flow rate of the first liquid phase is greater than that of the second liquid phase, the active component undergoes a rapid rebuffering from the acidic to neutral state of the precursor present in the second liquid phase. This in situ rebuffering of the active component is a particularly distinctive feature compared to conventional methods carried out in T- or Y-shaped mixers: in these conventional methods, a separate stabilization step is performed following the mixing, due to the difficulty of maintaining long-term stability of nanocarriers in the mixture environment. The method according to the present invention omits this step due to its in-situ rebuffering, which creates an environment in which nanocarriers are stable for a longer period of time directly in the active element. In conjunction with the diffusion process occurring in the active element, precursors present in ionized form are incorporated into the nanocarriers. In the case of LNPs, lipids briefly acquire a cationic charge upon contact with the second liquid phase (pH 4) and bind to anionic RNA or DNA from the second liquid phase. This mixture is then directly captured by the first liquid phase, and due to the higher buffering capacity of this medium, pH 7 prevails, and the particles are rapidly stabilized. If an active ingredient is present in the first or second liquid phase, these effects result in the active ingredient also being loaded onto the nanocarriers. Due to the large difference in flow rates in the active element, the nanocarriers / nanoformulations are finely distributed in the effluent, achieving a uniform particle size distribution in the range of only a few hundred nanometers.

[0055] The first and second liquid phases differ not only in their acidity but also in their chemical nature: the first liquid phase may be inorganic, while the second and / or third liquid phases are organic. Preferably, the first and second liquid phases are aqueous with different acidities, and the third liquid phase is organic. In this way, precursors and active ingredients soluble in different media can be used: for example, a water-soluble active ingredient can be supported on a carrier whose precursor is soluble only in organic solvents, or vice versa. Examples of water-soluble active ingredients include peptides, proteins, DNA, and RNA, particularly (m)RNA or siRNA. Lipids used as carriers (liposomes, LNPs) or polymers (particles, polyplexes) are dissolved in organic solvents, such as ethanol or acetonitrile. In such cases, the first dispersion medium is water, while the second dispersion medium is organic. In a three-component system, the first and second dispersion mediums are usually water, and the third dispersion medium is organic.

[0056] Suitable organic substances include, in particular, monohydric alcohols, such as ethanol, or polyhydric alcohols, such as glycerol, etc. Alternatively, acetonitrile or dimethyl sulfoxide (DMSO) can be used as organic substances.

[0057] An example of a precursor of nanocarriers soluble in organic solvents such as ethanol or glycerol is phosphatidylcholine (lecithin). Phosphatidylcholine can be obtained from eggs or soybeans. Further examples of precursors of nanocarriers soluble in organic solvents include cholesterol, 1,2-dioleyloxy-3-dimethylaminopropane (DODMA), polyethylene glycol dimethacrylate (PEG-DMA), and distearoylphosphatidylcholine (DSPC).

[0058] To adjust the pH, the first liquid phase may contain a buffering agent. This buffering agent ensures an environment in which the precursor is stable. The buffering agent may also be selected so that the acidity of the mixture, i.e., after mixing of the first and second liquid phases, has a desired value. This allows for corresponding control of the rebuffering step. If the first liquid phase is aqueous, the buffering agent may be selected from the group consisting of ammonium sulfate, acetate, polyvinyl alcohol, phosphate, and 2-(4-(2-hydroxyethyl)-1-piperazinyl)-ethanesulfonic acid.

[0059] In a preferred development of the method having at least one aqueous and organic phase, the first (aqueous) liquid phase contains a buffer, and the second (organic) liquid phase contains an active ingredient or a precursor of the active ingredient. Preferably, the buffer is polyvinyl alcohol and the organic substance is acetonitrile. The precursor of the nanocarrier selected is polylactide-co-glycolide, which is readily soluble in acetonitrile. However, it is also possible to dissolve the active ingredient in acetonitrile, for example, ritonavir. Instead of acetonitrile, acetone, dimethyl sulfoxide, or ethyl acetate can be used as an organic solvent.

[0060] Variations are also possible, such as a method in which the first (aqueous) liquid phase contains a buffer and the second (alcoholic) liquid phase contains two precursors of nanocarriers. Thus, for example, the buffer can be ammonium sulfate, while the two precursors of nanocarriers are lipid E ​​PC and cholesterol HP.

[0061] In a three-component system comprising a first aqueous phase, a second aqueous phase, and an organic phase, the second aqueous phase may also contain a species that is both a first precursor of the nanocarrier and a precursor of the active ingredient. The second precursor of the nanocarrier is then dissolved in the third (organic) phase. This configuration is used to produce nanoformulations in which LNPs are loaded with DNA or (m)RNA. Water-soluble DNA / (m)RNA is dissolved in the second aqueous phase under acidic conditions. Lipids are dissolved in the organic phase. These two are brought into contact in an active element, dispersed in the first liquid phase (aqueous and neutral), and simultaneously rebuffered. The DNA / (m)RNA is both the active ingredient and the precursor of the nanocarrier.

[0062] Regardless of the chemical nature of the liquid phase, the method can be carried out continuously if the discharge line of the device has the form of a circulation line and the first storage vessel simultaneously functions as a collection vessel. The method is managed so that the first liquid phase is first circulated before the second liquid phase is metered in. It is also possible to continuously withdraw the product while the first and optionally further liquid phases are circulated in the circulation line. At least the second liquid phase is then likewise continuously replenished. Thus, in this embodiment, the second liquid phase is supplied while at least the first phase is circulated. Preferably, the volumetric flow rate of the liquid circulating in the circulation line is greater than the amount of product withdrawn. The amount of replenished reactant preferably corresponds to the amount of product withdrawn, so that the total amount in the system is constant. The same applies to three-component systems.

[0063] The method according to the invention, made possible by the device according to the invention, results in a high-quality product, which is characterized in particular by a very uniform particle size distribution and, optionally, also by a uniform active ingredient concentration. The product of this method is a nanocarrier or nanoformulation.

[0064] The size of the resulting product is nanoscale, with an average particle size of less than 300 nm, preferably less than 200 nm, and particularly preferably between 40 nm and 140 nm. A specific type of nanocarrier always has a certain particle size. The particle size distribution is measured by the principle of dynamic light scattering using a Zetasizer, an instrument manufactured by Malvern Panalytical Ltd. (UK).

[0065] The resulting product has a polydispersity index of 0.08 to 0.2, which indicates uniform product quality. The polydispersity index (PDI) is measured by the principle of dynamic light scattering using a Zetasizer, an instrument manufactured by Malvern Panalytical Ltd. (UK). A PDI of 0.1 to 0.3 is particularly preferred.

[0066] This method allows the production of nanoformulations with high active ingredient concentrations. It is preferred if the mass concentration of the active ingredient in the nanoformulations according to the invention is between 1 ppm and 50%. This value is measured, for example, by HPLC-UV using an Agilent 1260 series instrument. Some active ingredients, such as mRNA, are administered in very low doses, resulting in a wide range of possible active ingredient concentrations.

[0067] This method also allows for the production of products with high active ingredient concentrations, as a high loading efficiency is achieved, meaning that a large proportion of the active ingredient provided is loaded onto the nanocarriers and not lost.

[0068] Surprisingly, it has been found that the device according to the invention for preparing nanocarriers and / or nanoformulations is also suitable for preparing microcarriers and / or microformulations.

[0069] In the context of the present invention, a microcarrier is a composition for carrying the active ingredients of pharmaceuticals, cosmetics, or dietary supplements. Microcarriers can be composed of a pure substance or a mixture of two or more substances. The substance can be solid, semi-liquid, or liquid. The substance can be a single-phase or multi-phase substance, and in either case, the microcarrier is particulate, with an average particle size greater than 1 μm and less than 1000 μm.

[0070] Thus, microcarriers within the meaning of the present invention are similar to nanocarriers except for size.

[0071] Examples of microcarriers are polymer particles with particle sizes ranging from 1 μm to 100 μm or 20 μm to 90 μm. In particular, biodegradable polymer particles in such particle size ranges are used as carriers for pharmaceuticals, nutritional supplements, or cosmetics.

[0072] Just as nanoformulations are based on nanocarriers, microformulations are also based on microcarriers.

[0073] Thus, in the context of the present invention, a microformulation is a dosage form of a pharmaceutical, cosmetic, or nutraceutical active ingredient carried on microcarriers, which may be on the surface of the microcarriers, inside the microcarriers, or complexed to the microcarriers.

[0074] The finding that the device according to the invention for preparing nanocarriers and / or nanoformulations is suitable for preparing microcarriers and / or microformulations is surprising, as the resulting particles are more than three times larger than nanoparticles.

[0075] Therefore, a further object of the present invention is the use of the device according to the invention for preparing microcarriers and / or microformulations.

[0076] In particular, further objects of the present invention are: 1. A method for producing microcarriers and / or microformulations, comprising: a) providing a device according to any one of claims 1 to 12; b) providing a first liquid phase containing a first liquid dispersion medium into a first storage vessel; c) providing a second liquid phase into a second storage container, the second liquid phase being a second liquid dispersion medium and containing at least one component selected from the group consisting of a precursor of the microcarrier, a precursor of the active ingredient, and the active ingredient; d) operating the pump to establish a liquid flow from the first storage container through the first supply line into the active element and through the discharge line into the collection container; e) metering a second liquid phase into the active element via a second supply channel, wherein the volumetric flow rate of the second liquid phase in the second supply channel is less than the volumetric flow rate of the liquid flow in the first supply channel; f) mixing the first and second liquid phases in the active element to obtain a mixture containing microcarriers and / or microformulations; g) collecting the mixture in a collection vessel; h) withdrawing the mixture from the fore-stage; i) optionally post-processing the mixture, in particular separating the microcarriers and / or microformulations from the mixture; The method includes:

[0077] The product of this method according to the invention is a microcarrier and / or a microformulation based thereon, with particles in the range of 1 μm to 1000 μm, preferably in the range of 1 μm to 100 μm, or 20 μm to 90 μm.

[0078] Preferred embodiments of the method according to the invention for preparing microcarriers and / or microformulations are similar to the preferred embodiments of the method according to the invention for preparing nanocarriers and / or nanoformulations.

[0079] The method according to the invention will now be described with reference to exemplary embodiments. [Brief explanation of the drawings]

[0080] [Figure 1] FIG. 1 shows a first variant of the device according to the invention; [Figure 2] FIG. 2 shows a second variant of the device according to the invention. [Figure 2x] 3 shows a second variant of the device according to the invention, showing details of the active elements and the supply channels; FIG. [Figure 2z] 3 shows a second variant of the device according to the invention, showing the section through the supply channel; FIG. [Figure 3] FIG. 3 shows a third variant of the device according to the invention. [Figure 4a] FIG. 1 is a schematic diagram showing a cross section of a vertical section of a two-component mixing device. [Figure 4b] FIG. 1 is a schematic diagram showing a cross section of a vertical section of a first modified example of a three-way mixing device. [Figure 4c] FIG. 10 is a schematic diagram showing a cross section of a vertical section of a second modified example of a three-way mixing device. [Figure 4d] FIG. 10 is a schematic diagram showing a cross section of a vertical section of a third modified example of a three-way mixing device. [Figure 5a] FIG. 2 shows the active element in a first axial position. [Figure 5b] FIG. 10 shows the active element in a second axial position. [Figure 6] FIG. 1 is a schematic diagram showing a Livitronic pump. [Figure 7a] FIG. 1 is a first cross-sectional view of the flow simulation. [Figure 7b] FIG. 12 is a second cross-sectional view of the flow simulation. [Figure 7c] FIG. 10 is a third cross-sectional view of the flow simulation. [Figure 8] FIG. 10 is a diagram showing an image of a flow vector.

[0081] FIG. 1 shows a schematic diagram of a first device 0 according to the invention. The device comprises a first storage container 1 in the form of a funnel-shaped tank and a second storage container 2 in the form of a plastic bag. The two storage containers 1 and 2 are used to contain a first and a second liquid phase. The volume of the first storage container 1 is significantly larger than the volume of the second storage container 2. The liquid phases are dispersions containing a liquid dispersion medium and precursors of nanocarriers and / or active ingredients, respectively.

[0082] The active element 3 is arranged below the storage vessels 1, 2 (vertically in the direction of acceleration due to gravity). The active element 3 serves to mix the two liquid phases. A first supply conduit 4 and a second supply conduit 5 connect the first storage vessel 1 / second storage vessel to the active element 3. The liquid phases can flow from the respective storage vessels 1, 2 through the respective supply channels 4, 5 to the active element 3. Both supply conduits 4, 5 extend vertically.

[0083] The pump 6 is arranged below the active element 3 (vertically in the direction of acceleration due to gravity). The pump 6 is used to circulate the mixture supplied by the active element 3. The active element 3 and the pump 6 are directly connected. The pump 6 may be rotatably driven by an integrated propulsion means 7.

[0084] Downstream of the pump 6 there is a discharge line 8 which extends horizontally for a predetermined distance before returning to the first storage vessel 1. The discharge line 8 is therefore configured as a circulation line 8+. The discharge line 8 is provided with a draw-off joint 9.

[0085] The entire apparatus 0 is housed in a frame 10. Control means and a metering device for metering the second liquid phase are not shown. The control means control the volumetric flow rate of the pump 6 and the volumetric flow rate of the metering device. The volumetric flow rate of the pump 6 corresponds to the volumetric flow rate of the mixture of the first and second liquid phases through the outlet 8, which is configured as a circulation path. In the mixture, the proportion of the first liquid phase is significantly higher than the proportion of the second liquid phase. The volumetric flow rate of the pump therefore corresponds to the volumetric flow rate of the first liquid phase (ignoring the second liquid phase). The pump is therefore effectively a kind of metering device for the first liquid phase.

[0086] During operation, the pump 6 first establishes a circuit for the first liquid phase from the first storage container 1 through the first conduit 4 into the active element 3 and back to the first storage container 1 through the outlet 8 / circulation 8+. A small amount of the second liquid phase from the second storage container 2 is then metered into the active element 3 via the second supply conduit 5. The two phases are mixed in the active element 3 to form a mixture. This mixture contains the dispersion medium and nanocarriers or nanocompounds dissolved therein. The nanocarriers or nanocompounds are formed by contact with the precursors in the active element 3.

[0087] The circuit is operated until the desired amount of product has accumulated in the mixture. The mixture is then removed through the withdrawal fitting 9 and the product is separated from the mixture. This is carried out outside the unit 0 using known separation devices. It is also possible to continuously remove the mixture from the circuit.

[0088] Figure 2 shows a ternary embodiment of the apparatus 0, which allows the production of a mixture from three liquid phases.

[0089] The first liquid phase is filled into a first storage vessel 1, which is also funnel-shaped and silo-like. The first storage vessel 1 therefore has the shape of a cone 11 on the inside. The angle of the cone 11 can be selected depending on the viscosity of the first liquid phase. In FIG. 2, the angle is 29°, and in FIG. 3, the angle is 55°, although angles up to 86° are also conceivable. The volume of the first storage vessel 1 is limited by an insert 111, which is also conical and has the same angle as the cone 11. The insert 111 can be mounted axially offset to vary the volume of the first storage vessel 1. This is particularly necessary when using the apparatus 0 for the production of various nanoformulations with different active ingredient concentrations. In recirculation operation, the residual amount in the first storage vessel 1 can also be adjusted via the insert 111. The first storage vessel 1 is also provided with two filling joints 1111, through which the storage vessel can be filled or replenished with the first liquid phase. The device 0 allows continuous operation by continuously supplying the first liquid phase via the filling joints 1111. At the bottom of the cone 11, the first storage vessel transforms into a first line 12, as can be seen in the enlarged view of FIG. 2x.

[0090] The storage vessels for the second and third liquid phases are not shown in Figure 2. However, second line 14 and third line 15 are shown, which serve as feed lines for the second liquid phase (via line 14) and for the third liquid phase (via line 15) entering active element 3.

[0091] Thus, the two conduits 14, 15 extend along the entire length of the device up to the active element 3. Between the bottom of the cone 11 and the active element 3, the second conduit 14 and the third conduit 15 extend vertically inside the first conduit 12. This region is called the vertical section, since all three conduits 12, 14, and 15 extend in the direction of acceleration due to gravity. Therefore, the flow vectors just before the contact of the three liquid phases in the active element are parallel to the gravity vector.

[0092] Furthermore, the conduits 14 and 15 also run parallel and vertically above the bottom of the cone 11, but this is not essential. Figure 2z shows a cross section of the second and third conduits 14, 15 in the vertical section. The shaded parts shown in Figure 2z are coatings.

[0093] At their respective openings in the active element 3, the two conduits 14 and 15 are each provided with a V-shaped grinding edge 13. This is more readily apparent in Figure 2x. The angle of the grinding edge 13 may be 15°, 30° or 60°.

[0094] It is also conceivable to pass the second liquid phase through both the second line 14 and the third line 15, in which case the second line and the third line each represent one conduit element of a duplicated second supply path of a two-phase mixing device for producing a biphasic mixture.

[0095] Figure 3 shows a third embodiment of the device 0 according to the invention, which corresponds substantially to the second variant shown in Figure 2. However, the volume of the first storage container 1 is different, being larger in the variant shown in Figure 3 than in Figure 2. The larger volume is achieved by arranging the insert 111 further up. This increases the space between the cone 11 of the first storage container and the corresponding conical part of the insert 111. Furthermore, the cone angle in the variant shown in Figure 3 is larger: it is now 55°, instead of only 29° in Figure 2.

[0096] Figures 4a and 4b show a cross section of a possible embodiment of the vertical section, i.e. a schematic view in the direction of acceleration due to gravity. All the conduits 12, 14, 15 are shown as circular, but they may also have corners.

[0097] The vertical section shown in Figure 4a is that of a two-phase device having a first conduit 12 for a first liquid phase and a second conduit 14 for a second liquid phase. The second conduit 14 extends inside the first conduit 12. Both conduits are arranged coaxially, i.e., their cross-sections have the same geometric center. The first liquid phase flows inside the first conduit 12 and outside the second conduit 14. The second liquid phase flows inside the second conduit 14. The arrangement shown in Figure 4a is also called a "single capillary".

[0098] The vertical section shown in FIG. 4b, called the "double capillary," is a three-way device having a first line 12 for a first liquid phase, a second line 14 for a second liquid phase, and a third line 15 for a third liquid phase. The second line 14 extends inside the first line 12, and the third line 15 also extends inside the first line 12. The second line 14 and the third line 15 are parallel. The first liquid phase flows inside the first line 12 and outside the second line 14 and the third line 15. The second liquid phase flows inside the second line 14. The third liquid phase flows inside the third line 15. It is also possible to pass the same second liquid phase through both the second line 14 and the third line 15. In that case, the device with the double capillary tube is used only in the two-component mode.

[0099] The vertical section shown in Figure 4c is that of a three-way mixer having a first pipe 12 for the first liquid phase, a second pipe 14 for the second liquid phase, and a third pipe 15 for the third liquid phase. The second pipe 14 extends inside the first pipe 12, and the third pipe 15 similarly extends inside the first pipe 12 and also inside the second pipe 14. All pipes 12, 14, and 15 are arranged coaxially with one another. The first liquid phase flows inside the first pipe 12 and outside the second pipe 14. The second liquid phase flows inside the second pipe 14 and outside the third pipe 15. The third liquid phase flows inside the third pipe 15.

[0100] The vertical section shown in Figure 4d is for a three-phase system with a first pipe 12 for the first liquid phase, a second pipe 14 for the second liquid phase, and a third pipe 15 for the third liquid phase. The second pipe 14 extends inside the first pipe 12, and the third pipe 15 also extends inside the first pipe 12. The second pipe 14 and the third pipe 15 are parallel. The first pipe 12 has an inner wall 16. Thus, the flow cross section of the first pipe is annular. The first liquid phase flows outside the inner wall 16 of the first pipe 12 and inside the outer wall of the first pipe 12, i.e., inside the shaded cross section at position 12. There is no material flow between the inner wall 16 and the walls of the second and third pipes 14 and 15. The second liquid phase flows in the second pipe 14. The third liquid phase flows in a third conduit 15 .

[0101] Figures 5a and 5b show how the second line 14, together with the third line 15, can be moved axially relative to the first line 12 in the three-way device shown in Figures 2 and 3. In Figure 5b, the vertical distance between the opening with the grinding end 13 and the pump 6 is greater than in Figure 5a, where the opening protrudes into the pump 6. This is achieved by a linear drive (not shown). The first line 12 does not move. The parallelism and vertical orientation of all lines 12, 14, and 15 are always maintained.

[0102] It is also conceivable that the second conduit 14 and the third conduit 15 can be moved axially independently of each other. This is not possible in the embodiment shown, as the second conduit 14 and the third conduit 15 can only be moved as a unit. Instead of moving the second and / or third conduits, the first conduit 12 can also be made axially movable, since the determining factor is not the absolute position of the conduits, but the relative axial position of the conduits. However, in the embodiment shown, the first conduit 12 is fixed.

[0103] By varying the axial position of the two conduits (12, 14) relative to one another, the flow conditions within the active element 3 are varied, thus optimizing the formation of nanocarriers / nanocompounds.

[0104] FIG. 5 is a schematic diagram of the pump 6. The pump 6 has the form of a centrifugal pump. The pump 6 has a stationary housing 17 in which a rotor 18 is rotatably mounted. The rotor 18 may also be called an impeller. The rotor 18 is propelled and mounted via a rotating magnetic field 7+. The axis of rotation of the rotor 18 is oriented vertically. The axis of rotation is preferably coaxial with the axis of the first conduit 12. This is the case for the variants shown in FIGS. 1, 2 and 3.

[0105] One feature of pump 6 is that it is a Livitronic pump, meaning that rotor 18 is magnetically coupled to propulsion means 7 via a rotating magnetic field 7+ rather than via a rigid shaft. The surrounding rotating magnetic field 7+ causes rotor 18 to rotate. This allows rotor 18 to be contactlessly mounted and to undergo rotational motion about its vertical axis of rotation.

[0106] In operation, the mixture enters the housing 17 of the pump 6 from above. The mixture is accelerated radially outward in a horizontal plane by the rotating rotor 18 and leaves the housing 16 again via the discharge channel 8. A portion of the mixture flows around the rotor 18 on its underside. This is possible due to the magnetic mounting and force transmission.

[0107] Figures 7a, 7b, and 7c show flow simulations through the active element 3 of the device shown in Figure 3. The figures show the local concentration of ethanol delivered through the capillary on the left. The "ethanol concentration" varies in all three spatial directions depending on the length of the capillary outside the lance. Each of the three cross-sections shows a cross-section approximately 2 mm above the end of the capillary. The first cross-section in Figure 7a shows that even with a path length of only approximately 2 mm, only a 1 mole percent ethanol concentration difference relative to the main flow is detectable. The second cross-section in Figure 7b (capillary length: 10 mm) shows the elongated core flow after 2 mm, with a concentration difference of approximately 6 mole percent. The third cross-section in Figure 7c (capillary length: 20 mm) shows the elongated core flow after 2 mm, with a concentration difference of approximately 10 mole percent.

[0108] Figure 8 shows a flow vector image of a device with two 3 mm capillaries in the area of ​​the active element. Here, the velocity vectors of the liquid flow are represented by small arrows. It is clear to those skilled in the art that the flow paths of the first liquid phase (outer) remain approximately parallel / vertical. Only in the area of ​​the feed paths of the second / third liquid phases via the two capillaries (center) is there a uniform deviation in the flow direction, if not turbulence. [Example]

[0109] The method according to the invention will now be explained in more detail with reference to experimental examples. Table 0 shows an overview of the examples.

[0110] All experiments were carried out using the apparatus corresponding to Figures 2 / 3. The change in the internal volume of the storage vessel 1 was carried out by displacing the insert 111. The apparatus had a Levitronic pump manufactured by Levitronix. For the metering of the first and second liquid phases, additional metering devices were used, which are not shown.

[0111] Polydispersity index (PDI) and average particle size (Zav) were measured using a Zetasizer from Malvern Panalytical Ltd, GB, with the backscattering angle set at 173°.

[0112] [Table 1-1] [Table 1-2]

[0113] Notes for Table 0: PVA = Polyvinyl alcohol EPC Chol = Lipid E ​​PC and Cholesterol HP NAT 8539 = Soy Phosphatidylcholine PLGA = poly(D,L-lactide-co-glycolide) Poly A = polyadenosine monophosphate DODMA = 1,2-dioleyloxy-3-dimethylaminopropane DSPC = distearoylphosphatidylcholine PEG-DMG = polyethylene glycol dimethacrylate PEI = polyethyleneimine

[0114] First group of embodiments: Production of liposomes as nanocarriers for pharmaceutical applications As shown in Examples I-III, the Livitronic pump allows for the robust production of liposomes over a wide range of lipid concentrations, where a ratio of Lipid E ​​PC and Cholesterol HP (55 mol%:45 mol%) was used as an example formulation at various concentrations.

[0115] Example I: In Example I, 700 mL of 250 mM ammonium sulfate buffer was initially charged as the first liquid phase, followed by 77.5 mL of the second liquid phase containing 200 mg / mL EPC-chol in ethanol. The steady volumetric flow rate of the first liquid phase was approximately 8 liters / minute, to which the second liquid phase was added at 10 mL / minute over approximately 8 minutes. The metering was performed using a rigid hose or capillary tube above the Livitronic pump. A liposome dispersion with an average particle size of approximately 150 nm was produced. The results were unchanged even when the metering rate was reduced by half and the batch size was reduced to 183.2 mL, demonstrating that the device is capable of good upscaling. The results are shown in Table 1.

[0116] Example II: In Example II, the lipid concentration in the organic phase was doubled while maintaining the same EPC-chol lipid ratio (55 mol%:45 mol%), and therefore the lipid concentration in the product was also doubled. Under comparable process parameters as in Example I, the resulting particles were larger and had higher polydispersity. In this example, the effect of the pump speed / constant volumetric flow of the first liquid phase was further investigated. For this formulation, a higher pump speed was shown to provide better results. The results are shown in Table 2.

[0117] Example III: In Example III, the lipid concentration in the organic phase was reduced while maintaining the same EPC-chol lipid ratio (55 mol%:45 mol%), and therefore the lipid concentration in the product. The lipid content in the organic phase was 15.2 mg / mL, and the aqueous to organic phase ratio was varied, while the effect of pump speed / constant volumetric flow of the first liquid phase was again investigated. A second capillary with a smaller inner diameter was also used. The results reflect a robust method, with results ranging from 79.85 nm to 108.4 nm. The smallest particle size was obtained using a fine capillary for metering the organic phase. The results are shown in Table 3.

[0118] Second Group of Embodiments: Liposome Carrier Systems for Cosmetic Applications Phospholipids occur naturally because they are important components of cell membranes. They are used as components of liposomes to carry low-solubility cosmetic active substances, such as vitamins A and E or coenzyme Q10, improving their penetration into the skin. These properties of liposomes have led to their increasing application in pharmaceutical formulations as well as cosmetics. Liposomes containing phospholipids also possess intrinsic biological activity, which is crucial for the normal functioning of the skin. Therefore, "empty" liposomes, which contain no contents, serve as moisturizers in cosmetics.

[0119] As shown in Examples IV and V, the Livitronic pump can be used to process phospholipids from a cosmetic standard formulation into liposomes, using NAT8539 (purified soybean phosphatidylcholine in ethanol) as an example formulation at various concentrations.

[0120] Example IV: In Example IV, liposomes were successfully produced by initially charging 583 g of water as the first liquid phase, followed by metering in 212 g of the second liquid phase, consisting of 107 g of NAT8539 (ethanol solution) and 105 g of ethanol. The metering was performed using a capillary tube above the Livitronic pump. The results of various combinations are shown in the table below. A higher metering rate tends to produce smaller particles. A higher flow rate (rpm) results in smaller particle sizes and a narrower particle size distribution. The effect of the distance between the capillary tube and the pump head is a negligible parameter. The results are shown in Table 4.

[0121] Example V: In Example V, liposomes were successfully produced by first charging 583 g of water as the first liquid phase, followed by metering in 212 g of a second liquid phase consisting of 53.5 g of NAT8539 (ethanol solution) and 158.5 g of ethanol. The lipid concentration in the organic phase was reduced by half, and therefore the lipid concentration in the product was also reduced by half. Metering was performed using a capillary tube above the Livitronic pump. The resulting particles were less than 200 nm in size and had a PDI below 0.2. The results are shown in Table 5.

[0122] Third Group of Embodiments: Biodegradable Nanoparticles as Drug Carrier Systems for Pharmaceutical Applications Poly(D,L-lactide-co-glycolide) (PLGA) is an established biodegradable polymer for the production of nanoparticles that can biodegrade in an aqueous environment to release active pharmaceutical ingredients over a controlled period of time. An example of a drug formulated in this way that has achieved significant commercial success is Eligard®, a treatment effective in treating symptoms resulting from prostate cancer. The active ingredient is leuprolide (a testosterone inhibitor) embedded in PLGA nanoparticles for subcutaneous injection.

[0123] Example VI: As shown in Example VI, the Livitronic pump allows for the production of a wide range of PLGA-based nanoparticles in a robust manner, with or without active ingredient loading. RESOMER® RG 502 H, a poly(D,L-lactide-co-glycolide) available from Evonik, was used as an example formulation in various mixing ratios with the aqueous phase (2% polyvinyl alcohol (PVA) solution). The PVA solution was initially charged as the first liquid phase and circulated at various pump speeds, while an organic PLGA solution in acetonitrile was metered in at various rates as the second liquid phase. For this purpose, RG 502 H was first completely dissolved in acetonitrile while stirring. The concentration of PLGA in the organic phase was 10 mg / mL.

[0124] They also successfully loaded PLGA nanoparticles with a model active ingredient with poor water solubility, namely ritonavir, which is approved for the treatment of HIV. In verum experiments, 10% or 20% ritonavir was added to the PLGA.

[0125] The resulting particles are suitable for sterile filtration and have a narrow particle size distribution, demonstrating their quality. It has been demonstrated that downstream processing by tangential flow filtration (TFF) to remove unencapsulated active ingredients and residual solvents, followed by lyophilization to achieve the required shelf life for pharmaceutical manufacturing, can be successfully performed. The resulting particles have an encapsulation efficiency of up to 14.59% and can be successfully redispersed after lyophilization, achieving particle size and particle size distribution similar to those immediately after the manufacturing process.

[0126] The results are shown in Table 6.

[0127] Fourth Group of Embodiments: Lipid Nanoparticles as Drug Carrier Systems for Pharmaceutical Use, Mainly for mRNA Since the FDA approved the first lipid nanoparticle-based formulation (Onpattro®) in 2018, interest in this formulation strategy has grown. Currently, LNPs are also being used to formulate mRNA as the active ingredient. This combines the general advantages of nanoparticle formulations (mechanical protection of the active ingredient, longer in vivo residence time, and preferential uptake in tumor tissue (EPR effect)) with the specific effects provided by the physicochemical properties of the lipids used. Thus, chemically unstable and charged mRNA can be transported across cell membranes by lipid nanoparticles, where the ionizable lipids at low pH ensure that the mRNA excretes its cargo and the information present in the mRNA can subsequently be translated into protein.

[0128] Example VII: As shown in Example VII, the livitronic pump allows for the production of lipid nanoparticles in a robust manner using polyadenosine monophosphate (polyA) as an alternative to mRNA.

[0129] A formulation containing DODMA (50 mol%) as an ionizable lipid, DSPC, cholesterol, and PEG-DMG (10 / 38.5 / 1.5 mol%) as additional components of the lipid fraction at a total lipid concentration of 15 mM was used as an example formulation. The volume of the lipid phase was 20 mL. The N / P ratio in the example formulation was 3:1. PolyA was dissolved in 60 mL of 5 mM acetate buffer at pH 4.0.

[0130] 70 mL of pH 7.4 phosphate buffer (USP) was added to the reservoir and circulated at 10,000 rpm using a Livitronic pump. Simultaneously, two capillaries were used to inject the acetate buffer containing poly(A) at 15 mL / min and the lipid solution at 5 mL / min. Upon collision of the solutions at the capillary outlet (see diagram), lipid nanoparticles formed at pH 4. These were rapidly contacted by the volumetric flow of the Livitronic pump and, due to the higher buffering capacity of the phosphate buffer, were adjusted to pH 7 and stabilized.

[0131] Results obtained by gel electrophoresis (E-Gel® EX Invitrogen Agarose 1%) show that polyA was successfully bound to the lipid nanoparticles, remained in the injection wells of the gel and produced a strong signal, while free polyA ran into the gel and showed a typical distribution.

[0132] The results are shown in Table 7.

[0133] Example VIII: An example formulation was used containing DODMA (50 mol%) as an ionizable lipid, DSPC, cholesterol, and PEG-DMG (10 / 38.5 / 1.5 mol%) as additional components of the lipid fraction at a total lipid concentration of 15 mM. The volume of the lipid phase was 37.5 mL. The N / P ratio in the example formulation was 3:1. PolyA was dissolved in 112.5 mL of 5 mM acetate buffer at pH 4.0, added to a reservoir, and circulated at 14,000 rpm using a Livitronic pump. Simultaneously, the lipid solution was injected at 5 mL / min through a capillary tube. Lipid nanoparticles were formed at pH 4 upon solution collision in the capillary outlet region. The particle solution was withdrawn from the device, and pH 7.4 phosphate buffer (USP) was added to externally adjust the pH to 7, stabilizing the particles.

[0134] The results are shown in Table 8.

[0135] Fifth Group of Embodiments: Polyplexes for Pharmaceutical Use, Primarily as Drug Carrier Systems for mRNA Similar to lipid nanoparticles (LNPs), polyplexes composed of, for example, polyethyleneimine and (m)RNA or DNA can be used to deliver these substances to their intracellular sites of action via non-viral delivery. Polyplexes can significantly enhance the stability of active ingredients. Polyplex formation occurs in weakly acidic solutions as a result of the charge difference between cationic polyethyleneimine and the anionic phosphate residues of (m)RNA or DNA.

[0136] Example IX: First, 80 mL of RNase-free water was filled into the device's reservoir and circulated at 10,000 rpm (first liquid phase). 33.5 mL of a 100 μg / mL polyA solution in RNase-free water was stirred to form the third liquid phase. This phase was injected through a capillary tube at 5 mL / min. 2.944 mL of a stock solution consisting of 10 g / L polyethyleneimine was mixed with 3 mL of 1 M acetate buffer and 29.056 mL of water (second liquid phase). The resulting N / P ratio was 15. This mixture was injected through a further capillary tube at 5 mL / min.

[0137] The results are shown in Table 9.

[0138] Example X: In Example X, the same amounts of polyA and polyethyleneimine were used as in Example IX. In this example, the polyA solution was diluted to a larger volume (115 mL) by introducing it into a first reservoir vessel and circulating it at 10,000 rpm. The polyethyleneimine solution (35 mL) described above in Example IX was injected through the capillary tube at 5 mL / min.

[0139] The results are shown in Table 10.

[0140] From the combination of Examples IX and X, those skilled in the art will recognize the good scalability of the method carried out by the device, since in Example X polyA was metered in via the first liquid phase at a much lower concentration compared to Example IX, where it was introduced via the third phase.

[0141] Sixth Group of Embodiments: Polymeric Micelles as Drug Carrier Systems for Pharmaceutical Use Polymeric micelles can be used as drug carrier systems that combine the systemic circulation-extending effect of polyethylene glycol (PEG) on the outer surface of the structure with the ability of polylactic acid (PLA) to carry (lipophilic) active ingredients within the micelle structure. Examples XI-XIII demonstrate polymeric micelle formulations in the nanometer size range produced by the mixing process described herein.

[0142] Example XI: First, 180 mL of deionized water was filled into the storage vessel of the device and circulated at 3000 rpm (first liquid phase). 16 mL of a stock solution consisting of 6.2% RESOMER® RP d 155, 55.8% ethanol, and 38.0% DMSO was used as the organic phase (second liquid phase). This mixture was infused through a capillary tube at 10 mL / min. RESOMER® RP d 155 is an mPEG-PLA diblock copolymer containing a 5 kDa mPEG block and 15 wt% PEG.

[0143] The results are shown in Table 11.

[0144] Example XII: First, 180 mL of deionized water was filled into the storage vessel of the device and circulated at 3000 rpm (first liquid phase). 16 mL of a stock solution consisting of 5.7% RESOMER® RP d 255, 51.3% ethanol, and 43.0% DMSO was used as the organic phase (second liquid phase). This mixture was infused through a capillary tube at 10 mL / min. RESOMER® RP d 255 is an mPEG-PLA diblock copolymer containing a 5 kDa mPEG block and 25 wt% PEG.

[0145] The results are shown in Table 12.

[0146] Example XIII: First, 180 mL of deionized water was filled into the storage vessel of the device and circulated at 3000 rpm (first liquid phase). 16 mL of a stock solution consisting of 7% RESOMER® RP d 505, 62.6% ethanol, and 30.4% DMSO was used as the organic phase (second liquid phase). This mixture was infused through a capillary tube at 10 mL / min. RESOMER® RP d 505 is an mPEG-PLA diblock copolymer containing a 5 kDa mPEG block and 50% PEG by weight.

[0147] The results are shown in Table 13.

[0148] Seventh Group of Embodiments: Biodegradable Microparticles as Drug Carrier Systems for Pharmaceutical Applications As shown in the third embodiment, mixing with a rivitronic pump allows for the production of PLGA-based nanoparticles over a wide concentration range in a robust manner, with or without active ingredient loading. By modifying process parameters and excipients, the method described herein can also be used to produce polymeric microparticles. A commercially available example of a successful polymeric microparticle is Bydureon® (Astra Zeneca). The method described herein can produce particles with a particle size range of 20 μm to 90 μm. Such particles are considered microcarriers.

[0149] RESOMER® RG 502 H, a poly(D,L-lactide-co-glycolide) available from Evonik, was used as an example formulation. Deionized water was filled as the first liquid phase and circulated at various pumping speeds. An organic PLGA solution was metered in as the second liquid phase (dispersed phase; referred to as DP) at various pumping speeds. To this end, RG 502 H was first completely dissolved in ethyl acetate while stirring. The PLGA concentration in the organic phase was varied between 5 and 30%. A 2% PVA solution was added as the third liquid phase (continuous phase; referred to as CP) via a second capillary tube. The CP / DP ratio was varied between 1:1 and 1:5.

[0150] In addition to the placebo formulation, we also successfully loaded a model active ingredient, meloxicam, an approved nonsteroidal anti-inflammatory drug, onto PLGA microparticles. In the verum experiments, meloxicam was dissolved in DMSO at 5% and added to the organic phase at 10% and 20% relative to the PLGA mass.

[0151] The resulting particles were filtered through a 20 μm sieve and resuspended in water. The microparticles were of suitable size, with a narrow size span indicating quality. The resulting meloxicam microparticles had an encapsulation efficiency of 92.7% (10% drug loading) to 96.6% (20% drug loading). As with all other examples, the size of the polymer microparticles was measured using a Mastersizer system (Malvern Panalytical).

[0152] The results are shown in Table 14.

[0153] [Table 2]

[0154] [Table 3]

[0155] [Table 4]

[0156] [Table 5-1] [Table 5-2]

[0157] [Table 6]

[0158] [Table 7-1] [Table 7-2]

[0159] [Table 8]

[0160] [Table 9]

[0161] [Table 10]

[0162] [Table 11]

[0163] [Table 12]

[0164] [Table 13]

[0165] [Table 14]

[0166] [Table 15-1] [Table 15-2] [Table 15-3] [Explanation of symbols]

[0167] 0 equipment 1. First storage container 2 Second storage container 3. Active Elements 4. First Supply Pipe 5 Second Supply Conduit 6. Pump 7 Propulsion means 7+ Rotating magnetic field 8 Exhaust channel 8+ circulation path 9 Extraction joint 10 frames 11 Cone 12 First Pipeline 13 Grinding end 14 Second Pipeline 15 Third Pipeline 16 Inner wall 17. Housing 18 rotors 111 Insert 1111 Filler joint

Claims

1. 1. An apparatus for producing nanocarriers and / or nanoformulations, comprising: a) a first storage vessel for containing a first liquid phase; b) a second storage vessel for containing a second liquid phase; c) an active element for mixing said first liquid phase with said second liquid phase to provide an at least two-phase mixture; d) a first supply line fluidly connecting said first reservoir to said active element; e) a second supply line fluidly connecting said second reservoir to said active element; f) a collection vessel for containing said mixture; g) a drainage passage fluidly connecting said active element to said collection vessel; h) a pump integrated in the discharge passage such that the mixture can be transported by the pump from the active element through the discharge passage into the collection vessel; 1. An apparatus having i) the first supply passage and the second supply passage are vertically oriented at least in a vertical section; j) in said vertical section, said second supply passage is disposed inside said first supply passage; k) in the vertical section, the first supply passage, or the second supply passage, or both supply passages are axially movable such that the axial position of the second supply passage relative to the first supply passage is adjustable. An apparatus comprising:

2. 2. The apparatus of claim 1, wherein the first supply passage in the vertical section is formed from a first linear conduit and the second supply passage in the vertical section is formed from a second linear conduit, the first linear conduit and the second linear conduit extending coaxially on the vertical section.

3. a third storage vessel for containing a third liquid phase, and a third supply line fluidly connecting the third storage vessel to the active element, the active element being configured to provide an at least three-phase mixture by mixing the first liquid phase with the second and third liquid phases, the first supply line in the vertical section being formed from a first linear conduit, the second supply line in the vertical section being formed from a second linear conduit, and the third supply line in the vertical section being formed from a third linear conduit. In the apparatus, a) the third supply channel is vertically oriented at least in the vertical section; b) in said vertical section, said third supply passage is disposed inside said first supply passage; c) in the vertical section, the third supply passage is axially movable such that the axial position of the third supply passage relative to the first supply passage is adjustable; 2. The device according to claim 1, characterized in that

4. 4. The apparatus of claim 3, wherein in said vertical section, said third supply passage is disposed within said second supply passage.

5. 5. Apparatus according to any one of claims 1 to 4, characterized in that the pump is in the form of a centrifugal pump having a housing and a rotor rotatably mounted within the housing about an axis of rotation and rotatably propellable via propulsion means.

6. 6. The device according to claim 5, characterized in that the axis of rotation of the rotor is oriented vertically and the horizontal section of the discharge passage extends in the plane of rotation of the rotor.

7. 6. The apparatus of claim 5, wherein said rotor is magnetically attached to said housing, and said propulsion means is a rotating magnetic field providing mechanically contactless power transmission from said propulsion means to said rotor.

8. 2. The apparatus of claim 1, wherein said recovery vessel and said first storage vessel are identical and said discharge passage is in the form of a circuit.

9. 5. The device according to claim 1, wherein the volumetric flow rate of the pump is adjustable.

10. 5. Apparatus according to claim 1, further comprising at least one metering device adapted for metering the second liquid phase to the active element.

11. 5. Apparatus according to claim 3 or 4, characterized in that it comprises at least one metering device arranged for metering the third liquid phase to the active element.

12. 11. Apparatus according to claim 10, characterized in that the volume flow rate of the metering device is adjustable.

13. 1. A method for producing a nanocarrier and / or nanoformulation, comprising: a) providing a device according to claim 1; b) providing a first liquid phase containing a first liquid dispersion medium into said first storage vessel; c) providing a second liquid phase in the second storage vessel, the second liquid phase being a second liquid dispersion medium and containing at least one component selected from the group consisting of a precursor of nanocarriers, a precursor of an active ingredient, and an active ingredient; d) operating the pump to establish a liquid flow from the first storage vessel through the first supply line into the active element and through the drain line into the collection vessel; e) metering the second liquid phase into the active element via the second supply line, the volumetric flow rate of the second liquid phase in the second supply line being less than the volumetric flow rate of the liquid phase in the first supply line; f) mixing said first liquid phase with said second liquid phase within said active element to obtain a mixture containing nanocarriers and / or nanoformulations; g) collecting the mixture in the collection vessel; h) withdrawing the mixture from the apparatus; i) optionally post-treating the mixture; A method comprising:

14. Providing an apparatus according to claim 3, a) providing a first liquid phase into the first storage vessel, the first liquid phase comprising a first liquid dispersion medium, the first liquid phase having a pH of 6-8; b) providing a second liquid phase in the second storage vessel, the second liquid phase comprising a second liquid dispersion medium and at least one precursor of the nanocarriers and / or the active ingredient, the second liquid phase having a pH of 3-5; c) providing a third liquid phase in the third storage vessel, the third liquid phase comprising a third liquid dispersion medium and at least one further component, the further component being selected from the group consisting of a precursor of nanocarriers, an active ingredient, and a precursor of an active ingredient; d) operating the pump to establish a liquid flow from the first storage vessel through the first supply line into the active element and through the drain line into the collection vessel; e) mixing the first, second and third liquid phases by metering the third liquid phase into the active element through the third supply line, wherein a volumetric flow rate of the third liquid phase in the third supply line is less than a volumetric flow rate of the liquid stream in the first supply line.

14. The method of claim 13, comprising:

15. 14. The method of claim 13, wherein the first liquid dispersion medium is water and the second liquid dispersion medium is an organic material.

16. 16. The method of claim 15, wherein the organic substance is a monohydric or polyhydric alcohol.

17. The method of claim 16, wherein the precursor of the nanocarrier is a phosphatidylcholine.

18. 16. The method of claim 15, wherein the first liquid phase contains a buffer.

19. 16. The method of claim 15, wherein the first liquid phase contains a buffer and the second liquid phase contains an active ingredient or a precursor of an active ingredient.

20. 16. The method of claim 15, wherein the first liquid phase contains a buffer and the second liquid phase contains two precursors of nanocarriers.

21. 14. A method according to claim 13, characterized in that it comprises providing an apparatus according to claim 8, and in that the first liquid phase is circulated before or during the metered addition of the second liquid phase.

22. 5. Use of a device according to any one of claims 1 to 4 for preparing microcarriers and / or microformulations.

23. A method for producing microcarriers and / or microformulations comprising: a) providing a device according to any one of claims 1 to 4; b) providing a first liquid phase containing a first liquid dispersion medium into said first storage vessel; c) providing a second liquid phase in the second storage vessel, the second liquid phase being a second liquid dispersion medium and containing at least one component selected from the group consisting of a precursor of the microcarriers, a precursor of the active ingredient, and the active ingredient; d) operating the pump to establish a liquid flow from the first storage vessel through the first supply line into the active element and through the drain line into the collection vessel; e) metering the second liquid phase into the active element via the second supply line, the volumetric flow rate of the second liquid phase in the second supply line being less than the volumetric flow rate of the liquid phase in the first supply line; f) mixing said first liquid phase with said second liquid phase within said active element to obtain a mixture containing microcarriers and / or microformulations; g) collecting the mixture in the collection vessel; h) withdrawing the mixture from the apparatus; i) optionally post-treating the mixture; A method comprising:

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