Apparatus and process for producing nanocarriers and/or nanoformulations

Abstract

An apparatus and corresponding process can be used for producing nanocarriers and or nanoformulations and corresponding process products. The apparatus is characterized by a vertical orientation of the feed conduits leading to active element. The feed conduits are nested within one another and are axially movable in terms of their orientation to one another. The process provides for the mixing of at least two liquid phases with different acidities. The volume flow of the first phase is greater than that of the second phase.

Claims

1. An apparatus for producing nanocarriers and/or nanoformulations, the apparatus comprising: a) a first reservoir vessel for accommodating a first liquid phase; b) a second reservoir vessel for accommodating a second liquid phase; c) an active element for providing an at least biphasic mixture by mixing the first liquid phase with the second liquid phase; d) a first feed by which the first reservoir vessel is in fluid communication with the active element; d) a second feed by which the second reservoir vessel is in fluid communication with the active element; f) a collection vessel for accommodating the mixture; g) a discharge by which the active element is in fluid communication with the collection vessel; h) a pump which is incorporated in the discharge in such a way that the mixture is conveyable by the pump from the active element via the discharge into the collection vessel; wherein the first feed and the second feed are vertically oriented at least on a vertical section; on the vertical section the second feed is arranged inside the first feed; and on the vertical section the first feed or the second feed or both feeds are axially displaceable such that an axial position of the second feed relative to the first feed is adjustable.

2. The apparatus as claimed in claim 1, wherein the first feed inside the vertical section is formed from a first linear pipeline, wherein the second feed inside the vertical section is formed from a second linear pipeline, and wherein the first linear pipeline and the second linear pipeline extend coaxially on the vertical section.

3. The apparatus as claimed in claim 1, further comprising a third reservoir vessel for accommodating a third liquid phase and a third feed by which the third reservoir vessel is in fluid communication with the active element, wherein the active element is adapted for providing an at least triphasic mixture by mixing the first liquid phase with the second liquid phase and the third liquid phase, and wherein the first feed inside the vertical section is formed from a first linear pipeline, the second feed inside the vertical section is formed from a second linear pipeline, and the third feed inside the vertical section is formed from a third linear pipeline, wherein a) the third feed is vertically oriented at least on the vertical section; b) on the vertical section the third feed is arranged inside the first feed; and c) the third feed is axially displaceable on the vertical section such that an axial position of the third feed relative to the first feed is adjustable.

4. The apparatus as claimed in claim 3, wherein on the vertical section the third feed is arranged inside the second feed.

5. The apparatus as claimed in claim 1, wherein the pump is in the form of a centrifugal pump which comprises a housing and a rotor which is rotatably mounted in the housing around a rotational axis and is rotatably propellable via a propulsion means.

6. The apparatus as claimed in claim 5, wherein the rotational axis of the rotor is vertically oriented and wherein a horizontal section of the discharge extends in a rotational plane of the rotor.

7. The apparatus as claimed in claim 5, wherein the rotor is magnetically mounted in the housing and wherein the propulsion means is a rotating field which allows mechanically contactless power transmission from the propulsion means to the rotor.

8. The apparatus as claimed in claim 1, wherein the collection vessel and the first reservoir vessel are identical and wherein the discharge is in the form of a circuit.

9. The apparatus as claimed in claim 1, wherein a volume flow of the pump is adjustable.

10. The apparatus as claimed in claim 1, further comprising at least one metered addition apparatus adapted for metered addition of the second liquid phase into the active element.

11. The apparatus as claimed in claim 3, further comprising at least one metered addition apparatus adapted for metered addition of the third liquid phase into the active element.

12. The apparatus as claimed in claim 10, wherein a volume flow of a metered addition apparatus is adjustable.

13. A process for producing a nanocarrier and/or a nanoformulation, the process comprising: a) providing the apparatus according to claim 1; b) providing the first liquid phase in the first reservoir vessel, wherein the first liquid phase contains a first liquid dispersion medium; c) providing the second liquid phase in the second reservoir vessel, wherein the second liquid phase is a second liquid dispersion medium and contains at least one component selected from the group consisting of a precursor to a nanocarrier, a precursor to an active ingredient, and an active ingredient; d) propelling the pump to establish a liquid flow from the first reservoir vessel via the first feed into the active element and via the discharge into the collection vessel; e) metering the second liquid phase via the second feed into the active element, wherein a volume flow of the second liquid phase in the second feed is smaller than a volume flow of the liquid flow in the first feed; f) mixing the first liquid phase and the second liquid phase in the active element to obtain a mixture containing a nanocarrier and/or a nanoformulation; g) collecting the mixture in the collection vessel; h) withdrawing the mixture from the apparatus; and i) optionally, working up the mixture.

14. The process as claimed in claim 13, wherein the apparatus further comprises a third reservoir vessel for accommodating a third liquid phase and a third feed by which the third reservoir vessel is in fluid communication with the active element, wherein the active element is adapted for providing an at least triphasic mixture by mixing the first liquid phase with the second liquid phase and the third liquid phase, and wherein the first feed inside the vertical section is formed from a first linear pipeline, the second feed inside the vertical section is formed from a second linear pipeline, and the third feed inside the vertical section is formed from a third linear pipeline, wherein the third feed is vertically oriented at least on the vertical section; on the vertical section the third feed is arranged inside the first feed; and the third feed is axially displaceable on the vertical section such that an axial position of the third feed relative to the first feed is adjustable; the process comprising: a) providing the first liquid phase in the first reservoir vessel, wherein the first liquid phase contains the first liquid dispersion medium and wherein the pH of the first liquid phase is between 6 and 8; b) providing the second liquid phase in the second reservoir vessel, wherein the second liquid phase contains the second liquid dispersion medium and at least one precursor to a nanocarrier and/or to an active ingredient, and wherein a pH of the second liquid phase is between 3 and 5; c) providing the third liquid phase in the third reservoir vessel, wherein the third liquid phase comprises a third liquid dispersion medium and at least one further component, wherein the further component is selected from the group consisting of a precursor to a nanocarrier, an active ingredient, and a precursor to an active ingredient; d) propelling the pump to establish a liquid flow from the first reservoir vessel via the first feed into the active element and via the discharge into the collection vessel; and e) mixing the first liquid phase and the second liquid phase and the third liquid phase by metering the third liquid phase via the third feed into the active element, wherein a volume flow of the third liquid phase in the third feed is smaller than the volume flow of the liquid flow in the first feed.

15. The process as claimed in claim 13, wherein the first liquid dispersion medium is water and the second liquid dispersion medium is an organic substance.

16. The process as claimed in claim 15, wherein the organic substance is a monohydric or polyhydric alcohol.

17. The process as claimed in claim 16, wherein the precursor to the nanocarrier is a phosphatidylcholine.

18. The process as claimed in claim 15, wherein the first liquid phase contains a buffer.

19. The process as claimed in claim 15, wherein the first liquid phase contains a buffer and wherein the second liquid phase contains the active ingredient or the precursor to the active ingredient.

20. The process as claimed in claim 15, wherein the first liquid phase contains a buffer and wherein the second liquid phase contains two precursors to a nanocarrier.

21. The process as claimed in claim 13, wherein the apparatus further comprises at least one metered addition apparatus adapted for metered addition of the second liquid phase into the active element, and wherein the first liquid phase is circulated before or during the metered addition of the second liquid phase.

22. A process for producing a microcarrier and/or a microformulation, the process, comprising the steps of: a) providing the apparatus according to claim 1; b) providing the first liquid phase in the first reservoir vessel, wherein the first liquid phase contains a first liquid dispersion medium; c) providing the second liquid phase in the second reservoir vessel, wherein the second liquid phase is a second liquid dispersion medium and contains at least one component selected from the group consisting of a precursor to a microcarrier, a precursor to an active ingredient, and an active ingredient; d) propelling the pump to establish a liquid flow from the first reservoir vessel via the first feed into the active element and via the discharge into the collection vessel; e) metering the second liquid phase via the second feed into the active element, wherein a volume flow of the second liquid phase in the second feed is smaller than a volume flow of the liquid flow in the first feed; f) mixing the first liquid phase and the second liquid phase in the active element to obtain a mixture containing a microcarrier and/or a microformulation; g) collecting the mixture in the collection vessel; h) withdrawing the mixture from the apparatus; and i) optionally, working up the mixture.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The processes according to the invention shall now be elucidated with reference to exemplary embodiments. To this end:

(2) FIG. 1: shows a first variant of an apparatus according to the invention

(3) FIG. 2A: shows a second variant of an apparatus according to the invention;

(4) FIG. 2B: shows a second variant of an apparatus according to the invention, showing the active element and feeds in detail;

(5) FIG. 2C: shows a second variant of an apparatus according to the invention, section through the feeds

(6) FIG. 3: shows a third variant of an apparatus according to the invention;

(7) FIG. 4A: shows a cross section of a vertical section of a binary apparatus, schematic view;

(8) FIG. 4B: shows a cross section of a vertical section of a 1st variant of a tertiary apparatus, schematic view;

(9) FIG. 4C: shows a cross section of a vertical section of a 2nd variant of a tertiary apparatus, schematic view;

(10) FIG. 4D: shows a cross section of a vertical section of a 3rd variant of a tertiary apparatus, schematic view;

(11) FIG. 5A: shows the active element in a first axial position;

(12) FIG. 5B: shows the active element in a second axial position;

(13) FIG. 6: shows a schematic diagram of a levitronic pump;

(14) FIG. 7A: shows a flow simulation, first sectional view;

(15) FIG. 7B: shows a flow simulation, second sectional view;

(16) FIG. 7C: shows a flow simulation, third sectional view;

(17) FIG. 8: shows a flow vector image.

DETAILED DESCRIPTION OF THE INVENTION

(18) FIG. 1 shows an overview of a first inventive apparatus 0. It comprises a first reservoir vessel 1 in the form of a funnel-shaped tank and a second reservoir vessel 2 in the form of a plastic bag. The two reservoir vessels 1, 2 are used for accommodating a first and second liquid phase. The volume of the first reservoir vessel 1 is markedly greater than the volume of the second reservoir vessel 2.

(19) The liquid phases are each dispersions containing a liquid dispersion medium and precursors to nanocarrier and/or active ingredients.

(20) An active element 3 is arranged below the reservoir 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 feed conduit 4 and a second feed conduit 5 connect the first reservoir vessel 1/the second reservoir vessel with the active element 3. The liquid phases can flow from their respective reservoir vessels 1, 2 through the respective feed 4, 5 to the active element 3. Both feed conduits 4, 5 extend vertically.

(21) A 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 provided by the active element 3. The active element 3 and the pump 6 are directly connected. The pump 6 may be rotatably propelled by an integrated propulsion means 7.

(22) Provided downstream of the pump 6 is a discharge 8 which extends horizontally for a distance before returning to the first reservoir vessel 1. The discharge 8 is therefore configured as circuit 8+. The discharge 8 is provided with a withdrawal fitting 9.

(23) The entire apparatus 0 is accommodated in a frame 10. A control means and a metered addition apparatus for metered addition of the second liquid phase are not shown. The control means controls the volume flow of the pump 6 and the volume flow of the metered addition apparatus. The volume flow of the pump 6 corresponds to the volume flow of the mixture of the first and second liquid phase through the discharge 8 configured as a circuit. In the mixture the proportion of the first liquid phase is markedly greater than the proportion of the second liquid phase. The volume flow of the pump thus corresponds to the volume flow of the first liquid phase (neglecting the second liquid phase). The pump is thus effectively a type of metered addition apparatus for the first liquid phase.

(24) In operation the pump 6 initially establishes a circuit of the first liquid phase from the first reservoir vessel 1, through the first conduit 4 into the active element 3 and through the discharge 8/circuit 8+ back into the first reservoir vessel 1. A small amount of the second liquid phase from the second reservoir vessel 2 is then metered into the active element 3 via the second feed conduit 5. The two phases are mixed in the active element 3 to form a mixture. The mixture contains dispersion medium and, dissolved therein, nanocarrier or nanoformulation which is formed by contact of the precursors in the active element 3.

(25) The circuit is operated until a desired amount of product has accumulated in the mixture. The mixture is then withdrawn via the withdrawal fitting 9 and the product separated from the mixture. This is carried out outside the apparatus 0 with known separation apparatuses. It is also possible to withdraw the mixture from the circuit continuously.

(26) FIG. 2A shows a tertiary embodiment of the apparatus 0. This allows production of mixtures from three liquid phases

(27) The first liquid phase is filled into the first reservoir vessel 1 which is again funnel-shaped, like a silo. Accordingly, the first reservoir vessel has the shape of a cone 11 on the inside. The angle of the cone 11 may be selected according to the viscosity of the first liquid phase. In FIG. 2A the angle is 29? and in FIG. 3 the angle is 55? but angles up to 86? are also conceivable. The volume of the first reservoir vessel 1 is limited by an insert 111. The insert 111 is likewise conical and has the same angle as cone 11. The insert 111 may be installed axially offset to vary the volume of the first reservoir vessel 1. This is especially necessary when the apparatus 0 is used for production of different nanoformulations which differ in their active ingredient concentration. In recirculating operation the holdup of the first reservoir vessel may also be adjusted via the insert 111. The first reservoir vessel 1 is also provided with two charging fittings 1111 through which the reservoir vessel may be filled or replenished with first liquid phase. The apparatus 0 allows continuous operation by continuously supplying the first liquid phase via the charging fittings 1111. At the foot of the cone 11 the first reservoir vessel transitions into a first pipeline 12. This is readily apparent in the magnified view of FIG. 2B.

(28) The reservoir vessels for the second and third liquid phase are not shown in FIG. 2A. However, a second pipeline 14 and a third pipeline 15, which serve as a feed for second liquid phase (via pipeline 14) and third liquid phase (via pipeline 15) into the active element 3, are shown.

(29) Accordingly, the two pipelines 14 and 15 extend along the full length of the apparatus to the active element 3. Between the foot of the cone 11 and the active element 3 the second pipeline 14 and the third pipeline 15 extend vertically and inside the first pipeline 12. This region is referred to as the vertical section because all three pipelines 12, 14 and 15 extend in the direction of acceleration due to gravity. The flow vectors of the three liquid phases immediately before contact thereof in the active element is thus parallel to the gravitational vector.

(30) The pipelines 14 and 15 moreover also run parallel and vertical above the foot of the cone 11 but this is not relevant. FIG. 2C shows a cross section through the second and third pipelines 14, 15 above the vertical section. The shaded component shown in FIG. 2C is a sheathing.

(31) At their respective openings in active element 3 the two pipelines 14 and 15 have each been provided with a V-shaped ground end 13. This is more easily apparent in FIG. 2B. The angle of the ground end 13 may be 15? or 30? or 60?.

(32) It is also conceivable to pass second liquid phase through both the second pipeline 14 and the third pipeline 15. The second and the third pipeline are then each one conduit element of a dualized second feed conduit of a binary apparatus for producing a biphasic mixture.

(33) FIG. 3 shows a third inventive embodiment of the apparatus 0. It corresponds substantially to the second variant shown in FIG. 2A. However, the difference is in the volume of the first reservoir vessel 1 which is greater in the variant shown in FIG. 3 than in FIG. 2A. The greater volume has been established by positioning the insert 111 further upwards. This enlarges the space between the cone 11 of the first reservoir vessel and the corresponding cone of the insert 111. Furthermore, the cone angle in the variant shown in FIG. 3 is greater; it is 55? here instead of only 29? in FIG. 2A.

(34) FIGS. 4A to 4B show a schematic view of possible embodiments of the vertical section in cross section, i.e. in the perspective of acceleration due to gravity. All pipelines 12, 14, 15 are shown as circular but may also be angular.

(35) The vertical section shown in FIG. 4A belongs to a binary apparatus having a first pipeline 12 for the first liquid phase and a second pipeline 14 for the second liquid phase. The second pipeline 14 extends inside the first pipeline 12. Both pipelines are arranged coaxially, i.e. the geometric center of their cross-sections is identical. The first liquid phase flows inside the first pipeline 12 and outside the second pipeline 14. The second liquid phase flows inside the second pipeline 14. The constellation shown in FIG. 4A is also referred to as single capillary.

(36) The vertical section shown in FIG. 4B referred to as dual capillary belongs to a tertiary apparatus having a first pipeline 12 for the first liquid phase, a second pipeline 14 for the second liquid phase and a third pipeline 15 for the third liquid phase. The second pipeline 14 extends inside the first pipeline 12 and the third pipeline 15 likewise extends inside the first pipeline 12. The second pipeline 14 and the third pipeline 15 are parallel. The first liquid phase flows inside the first pipeline 12 and outside the second pipeline 14 and the third pipeline 15. The second liquid phase flows inside the second pipeline 14. The third liquid phase flows inside the third pipeline 15. It is also conceivable to pass the same second liquid phase through both the second pipeline 14 and the third pipeline 15. The apparatus with the dual capillary is then utilized merely in binary fashion.

(37) The vertical section shown in FIG. 4C belongs to a tertiary apparatus having a first pipeline 12 for the first liquid phase, a second pipeline 14 for the second liquid phase and a third pipeline for the third liquid phase. The second pipeline 14 extends inside the first pipeline 12 and the third pipeline 15 likewise extends inside the first pipeline 12 and, in addition, also inside the second pipeline 14. All pipelines 12, 14, 15 are arranged coaxially to one another. The first liquid phase flows inside the first pipeline 12 and outside the second pipeline 14. The second liquid phase flows inside the second pipeline 14 and outside the third pipeline 15. The third liquid phase flows inside the third pipeline 15.

(38) The vertical section shown in FIG. 4D belongs to a tertiary apparatus having a first pipeline 12 for the first liquid phase, a second pipeline 14 for the second liquid phase and a third pipeline for the third liquid phase. The second pipeline 14 extends inside the first pipeline 12 and the third pipeline 15 likewise extends inside the first pipeline 12. The second pipeline 14 and the third pipeline 15 are parallel. The first pipeline 12 here comprises an inner wall 16. The flow cross section of the first pipeline is therefore circular. The first liquid phase flows outside the inner wall 16 of the first pipeline 12 and inside the outer wall of the first pipeline 12, i.e. the shaded sectional area with position 12. No material flow takes place between the inner wall 16 and the wall of the second/third pipeline 14, 15. The second liquid phase flows inside the second pipeline 14. The third liquid phase flows inside the third pipeline 15.

(39) FIGS. 5A and 5B show how in the tertiary apparatuses shown in FIGS. 2A and 3 the second pipeline 14 together with the third pipeline 15 may be axially displaced relative to the first pipeline 12. In FIG. 5B the vertical distance between the opening with the ground end 13 and the pump 6 is greater than in FIG. 5A where the opening projects into the pump 6. This is accomplished with a linear drive not shown here. The first pipeline 12 remains unmoved. The parallel nature and vertical orientation of all pipelines 12, 14, 15 is always retained.

(40) It is also conceivable to make the second pipeline 14 and the third pipeline 15 axially movable separately from one another. This is not possible in the embodiments shown here; the second pipeline 14 and the third pipeline 15 can only be moved as a package. It is also possible to make the first pipeline 12 axially movable as an alternative to moving the second and/or third pipeline. This is because the decisive factor is the relative axial orientation of the pipelines and not the absolute position. However, in the embodiments shown here the first pipeline 12 is fixed.

(41) By varying the axial orientation of the two pipelines 12, 14 to one another the flow conditions in the active element 3 are varied and the formation of the nanocarriers/the nanoformulation thus optimized.

(42) FIG. 6 is a schematic diagram of the pump 6. The pump 6 is in the form of a centrifugal pump. It comprises an immovable housing 17 in which a rotor 18 is rotatably mounted. The rotor 18 may also be referred to as an impeller. The propulsion and mounting of the rotor 18 is effected via a rotating field 7*. The rotational axis of the rotor 18 is vertically oriented. It is preferable when the rotational axis is coaxial with the axis of the first pipeline 12. This is the case in the variants shown in FIGS. 1, 2 and 3.

(43) One feature of the pump 6 is that it is a levitronic pump. This means that the rotor 18 is coupled to the propulsion means 7 not via a rigid shaft but rather magnetically via the rotating field 7*. The surrounding rotating field 7* rotates the rotor 18. This allows the rotor 18 to be contactlessly mounted and set into rotary motion around its vertical rotational axis.

(44) In operation the mixture enters the housing 17 of the pump 6 from above. The mixture is accelerated radially outwards in a horizontal plane by the rotating rotor 18 and exits the housing 16 again via the discharge 8. A portion of the mixture flows around the rotor 18 on its underside. This is possible on account of the magnetic mounting and force transmission.

(45) FIGS. 7A, 7B, and 7C show a flow simulation through an active element 3 of the apparatus shown in FIG. 3. The figure shows the local concentration of ethanol supplied via the left hand capillary. The ethanol concentration varies in all three spatial directions according to the capillary length outside the lance. The three sectional views show the cross section in each case about 2 mm beyond the end of the capillaries. The first sectional view in FIG. 7A shows that even after a path length of only about 2 mm only a 1 mol percent concentration difference of ethanol relative to the main flow is detectable. The second sectional view (capillary length 10 mm) shown in FIG. 7B shows after 2 mm an elongate core flow where the concentration difference is about 6 mol percent. The third sectional view (capillary length 20 mm) shown in FIG. 7C shows after 2 mm an elongate core flow where the concentration difference is about 10 mol percent.

(46) FIG. 8 shows a flow vector image of an apparatus having two 3 mm capillaries in the region of the active element. The velocity vectors of the liquid flow are represented therein as small arrows. It is apparent to those skilled in the art that the flow paths of the first liquid phase (outside) remain largely parallel/vertical. Only in the region of the feed for the second/third liquid phase (central) via the two capillaries is there a homogenous deviation of the flow direction, though this cannot be described as turbulent.

EXAMPLES

(47) The processes according to the invention will now be more particularly elucidated with reference to experimental examples. Table 0 contains an overview of the examples.

(48) All experiments were carried out with an apparatus corresponding to FIGS. 2A/3. The internal volume of the reservoir 1 was varied by moving the insert 111. The apparatus comprised a levitronic pump from Levitronix. Metered addition apparatuses were additionally employed for metered addition of the first and second liquid phase. These are not shown in the drawings.

(49) A Zetasizer instrument from Malvern Panalytical Ltd, GB was used to determine polydispersity (PDI) and average particle size (Zav). The backscattering angle was set to 173?.

(50) TABLE-US-00001 TABLE 0 Overview of examples Third liquid phase First liquid phase Second liquid phase Nanocarrier/ Example Dispersion Active Dispersion Nanocarrier Active Dispersion active ingredient Group no. medium Buffer ingredient medium Buffer precursor ingredient medium precursor 1 I Water Ammonium n/a Ethanol n/a EPC chol n/a n/a n/a sulphate 1 II Water Ammonium n/a Ethanol n/a EPC chol n/a n/a n/a sulphate 1 III Water Ammonium n/a Ethanol n/a EPC chol n/a n/a n/a sulphate 2 IV Water n/a n/a Ethanol n/a NAT 8539 n/a n/a n/a 2 V Water n/a n/a Ethanol n/a NAT 8539 n/a n/a n/a 3 VI Water PVA n/a Acetonitrile n/a PLGA Ritonavir n/a n/a 4 VII Water Phosphate n/a Water Acetate Poly A Poly A Ethanol DODMA, DSPC, Cholesterol, PEG- DMG 4 VIII Water Phosphate n/a Water Acetate Poly A Poly A Ethanol DODMA, DSPC, Cholesterol, PEG- DMG 5 IX Water n/a n/a Water Acetate PEI n/a Water Poly A 5 X Water n/a Poly A Water Acetate PEI n/a n/a n/a 6 XI Water n/a n/a Ethanol/DMSO n/a RESOMER? n/a n/a n/a RP d 155 6 XII Water n/a n/a Ethanol/DMSO n/a RESOMER? n/a n/a n/a RP d 255 6 XIII Water n/a n/a Ethanol/DMSO n/a RESOMER? n/a n/a n/a RP d 505 7 XIV Water n/a n/a Ethyl acetate n/a PLGA Meloxicam PVA n/a (in DMSO) Legend for table 0: PVA = polyvinyl alcohol EPC Chol = lipoid 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

Example Series 1: Production of Liposomes as Nanocarriers for Pharma Applications

(51) As shown in examples I to III the levitronic pump allows generation of liposomes over a wide concentration range of lipids with a robust process. Lipoid E PC and cholesterol HP in the ratio (55 mol %:45 mol %) were used here in varying concentrations as an example formulation.

Example I

(52) In example I 700 mL of a 250 mM ammonium sulfate buffer was initially charged as first liquid phase and 77.5 mL of second liquid phase comprising 200 mg/ml of EPC-chol in ethanol were metered in. The stationary volume flow of the first liquid phase was around 8 liters per minute, into which 10 ml per minute of second liquid phase were then added over a period of around 8 minutes. The metered addition was carried out using a rigid hose or a capillary above the levitronic pump. A liposome dispersion having an average particle size of about 150 nm was produced. The result was also reproducible at half the metered addition rate and a batch size reduced to 183.2 ml. This demonstrates that the apparatus allows good upscaling. Results are reported in table 1.

Example II

(53) In example II the lipid concentration in the organic phase and thus also in the product was doubled while retaining the same EPC-chol lipid ratio (55 mol %:45 mol %). At comparable process parameters to those in example I the resulting particles are larger and have a higher polydispersity. This example further investigated the effect of pump speed/stationary volume flow of the first liquid phase. It was shown that for this formulation higher pump rates give better results. Results are reported in table 2.

Example III

(54) In example III the lipid concentration in the organic phase and thus also in the product was reduced while retaining the same EPC-chol lipid ratio (55 mol %:45 mol %). The lipid content and the organic phase was 15.2 mg/ml and the ratio of aqueous phase to organic phase was varied while the effect of pump speed/stationary volume flow of the first liquid phase was once again investigated. A second capillary of reduced internal diameter was also employed. The results reflect a robust process with results between 79.85 nm and 108.4 nm. The lowest particle size was achieved with a fine capillary for metered addition of the organic phase. Results are reported in table 3.

Example Series 2: Liposomal Carrier Systems for Cosmetic Applications

(55) Phospholipids are important constituents of the cell membranes and thus naturally occurring. They may be used as components of liposomes as carriers for low-solubility cosmetically active substances such as for example Vitamin A & E or Coenzyme Q10 and improve skin penetration. These properties of the liposomes are finding increasing application in cosmetics as well as the formulation of pharmaceuticals. The liposomes containing phospholipids also have intrinsic biological activities, and this is of great importance for the normal functioning of the skin. Accordingly, empty liposomes without any contents have their place as moisturizers in cosmetics.

(56) As shown in examples IV and V the levitronic pump may be used to process phospholipids of a cosmetic standard formulation into liposomes. NAT 8539 (purified soy phosphatidylcholine in ethanol) was employed here in varying concentration as an example formulation.

Example IV

(57) In example IV successful production of liposomes was achieved by initially charging 583 g of water as first liquid phase, followed by metered addition of 212 g of second liquid phase consisting of 107 g of NAT 8539 (ethanolic solution) and 105 g ethanol. The metered addition was carried out using a capillary above the levitronic pump. The results of various arrangements are shown in the table which follows. A higher rate of metered addition tends to result in smaller particles. Higher flow rates (rpm) result in smaller particle sizes and narrower particle size distribution. The effect of the distance between the capillary and the pump head is a parameter that may be neglected. The results are shown in table 4.

Example V

(58) In example V successful production of liposomes was achieved by initially charging 583 g of water as first liquid phase, followed by metered addition of 212 g of second liquid phase consisting of 53.5 g of NAT 8539 (ethanolic solution) and 158.5 g ethanol. The lipid concentration in the organic phase and thus also in the product was thus halved. The metered addition was carried out using a capillary above the levitronic pump. The resulting particles are <200 nm and have a PDI below 0.2. The results are shown in table 5.

Example Series 3: Biodegradable Nanoparticles as Pharmaceutical Carrier Systems for Pharmaceutical Applications

(59) Poly(D,L-lactide-co-glycolide) (PLGA) is an established biodegradable polymer for producing nanoparticles which biodegrade in an aqueous environment over controllable time periods to allow release of an active pharmaceutical ingredient. An example formulated in this way which is very successful on the market is Eligard?, an effective therapy for treatment of symptoms occurring due to prostate cancer. The active ingredient is Leuprolid (a testosterone inhibitor) embedded in PLGA nanoparticles for subcutaneous injection.

Example VI

(60) As shown in example VI the levitronic pump allows PLGA-based nanoparticles to be produced over a wide concentration range with a robust process with or without active ingredient loading. RESOMER? RG 502 H, a poly(D,L-lactide-co-glycolide), obtainable from Evonik was employed as an example formulation in a varying mixture ratio to the aqueous phase (2% polyvinyl alcohol (PVA) solution). The PVA solution was initially charged as first liquid phase and circulated at varying rates (pump speeds) and the organic PLGA solution in acetonitrile was metered in at varying rates as a second liquid phase. To this end, RG 502 H was first completely dissolved in acetonitrile with stirring. The concentration of the PLGA in the organic phase was 10 mg/ml.

(61) The PLGA nanoparticles were also successfully loaded with a model active ingredient having poor solubility in water, namely Ritonavir which is approved for treatment of HIV. In the verum experiments 10% or 20% of Ritonavir based on the PLGA were additionally added.

(62) The resulting particles are suitable in terms of size for sterile filtration and have a narrow particle size distribution which is indicative of quality. It was demonstrated that downstream processing by tangential flow filtration (TFF) to remove unencapsulated active ingredient and residual solvent and freeze-drying to achieve a required shelflife, as required for pharmaceutical production, may be carried out successfully. The obtained particles have an encapsulation efficiency of up to 14.59% and are successfully redispersible after freeze-drying to achieve similar particle sizes and particles size distributions as immediately after the production process.

(63) The results are shown in table 6.

Example Series 4: Lipid Nanoparticles as Pharmaceutical Carrier Systems for Pharmaceutical Applications, Primarily mRNA

(64) Since the approval of a first preparation based on lipid nanoparticles (Onpattro?) by the FDA in 2018 there has been increasing interest in this formulation strategy. LNPs are today also used for formulation of mRNA as active constituents. This combines the advantages of nanoparticulate formulations generally (mechanical protection of the active ingredient, longer residence time in the organism, preferential uptake in tumor tissue (EPR effect)) with specific effects brought about by the physicochemical properties of the employed lipid. Accordingly, the chemically unstable and charged mRNA may be transported through the cell membrane with a lipid nanoparticle, where at low pH ionizable lipids ensure that the mRNA discharge cargo and the information present in the mRNA can later be translated into proteins.

Example VII

(65) As shown in example VII the levitronic pump allows lipid nanoparticles to be produced with a robust process with polyadenosine monophosphate (poly A) as a surrogate for mRNA.

(66) A formulation containing DODMA (50 mol %) as an ionizable lipid and DSPC, cholesterol and PEG-DMG (10/38.5/1.5 mol %) as further constituents of the lipid fraction at a combined lipid concentration of 15 mM was used as an example formulation. The volume of the lipid phase was 20 ml. The N/P ratio was 3:1 in the example formulation. The poly A was dissolved in 60 ml of a 5 mM acetate buffer at pH 4.0.

(67) 70 mL of a phosphate buffer pH 7.4 USP were added to the reservoir vessel and circulated with the levitronic pump at 10 000 rpm. Simultaneously, the poly A-containing acetate buffer was injected at 15 ml/min and the lipid solution at 5 ml/min using two capillaries. Upon impacting of the solutions in the region of the capillary outlet (see diagram) lipid nanoparticles were formed at pH 4 and these were immediately contacted with the phosphate buffer due the volume flow of the levitronic pump and, as a result of its higher buffer capacity, adjusted to pH 7 and thus stabilized.

(68) Results obtained by gel electrophoresis (E-Gel? EX invitrogen Agarose 1%) show that the poly A was successfully bound in the lipid nanoparticles, remained in the injection well of the gel and produced a strong signal while free Poly A ran into the gel and showed a typical distribution.

(69) The results are shown in table 7.

Example VIII

(70) A formulation containing DODMA (50 mol %) as an ionizable lipid and DSPC, cholesterol and PEG-DMG (10/38.5/1.5 mol %) as further constituents of the lipid fraction at a combined lipid concentration of 15 mM was used as an example formulation. The volume of the lipid phase was 37.5 ml. The N/P ratio was 3:1 in the example formulation. The poly A was dissolved in 112.5 ml of a 5 nnM acetate buffer at pH 4.0 and added to the reservoir vessel and circulated with the levitronic pump at 14 000 rpm. Simultaneously, the lipid solution was injected at 5 ml/min using a capillary. Upon impacting of the solutions in the region of the capillary outlet lipid nanoparticles were formed at pH 4. The particle solution was withdrawn from the apparatus and externally adjusted to pH 7 by addition of phosphate buffer pH 7.4 USP to stabilize the particles.

(71) The results are shown in table 8.

Example Series 5: Polyplexes as Pharmaceutical Carrier Systems for Pharmaceutical Applications, Primarily mRNA

(72) Similarly to the lipid nanoparticles (LNPs) it is also possible to use polyplexes, for example composed of polyethyleneimine and (m)RNA or DNA, to bring these substances to their intracellular site of action by non-viral delivery. Polyplexes can markedly increase the stability of the active constituents. The formation of the polyplexes is a consequence of charge differences between the polyethyleneimine which is cationic in slightly acidic solution and the anionic phosphate residues of the (m)RNA or DNA.

Example IX

(73) 80 mL of RNase-free water were initially charged in the reservoir of the apparatus and circulated at 10 000 rpm (first liquid phase). 33.5 mL of a 100 ?g/ml poly A solution in RNase-free water were produced with stirring and serve as third liquid phase. This phase was injected via a capillary at 5 ml/min. 2.944 ml of a stock solution consisting of 10 g/l of polyethyleneimine are mixed with 3 mL of a 1M acetate buffer and 29.056 ml of water (second liquid phase). The resulting N/P ratio is 15. This mixture is injected at 5 ml/min via a further capillary.

(74) The results are shown in table 9.

Example X

(75) In example X identical amounts of poly A and polyethyleneimine as in example IX were employed but in this example the poly A solution was introduced into the first reservoir diluted in a greater volume (115 ml) and circulated at 10 000 rpm. The polyethyleneimine solution previously described in example IX (35 mL) is injected at 5 ml/min via a capillary.

(76) The results are shown in table 10.

(77) From a combination of the examples IX and X those skilled in the art will recognize the good scalability of the process operated with the apparatus, since in example X poly A was metered in via the first liquid phase in a much lower concentration than in example IX where poly A was introduced via the third phase.

Example Series 6: Polymeric Micelles as Pharmaceutical Carrier Systems for Pharmaceutical Applications

(78) Polymeric micelles can be used as pharmaceutical carrier systems, in which the systemic circulation prolonging effect of polyethylene glycol (PEG) on the outer surface of the structure is combined with the ability of polylactic acid (PLA) to load (lipophilic) active components within the micellar structure. Example XI to XIII show polymeric micelles formulations in the nanometer size range produced by the herein described mixing process.

Example XI

(79) 180 mL of deionized water were initially charged in the reservoir of the apparatus and circulated at 3 000 rpm (first liquid phase). 16 mL of a stock solution consisting of 6.2% RESOMER? RP d 155, 55.8% ethanol, 38.0% DMSO served as organic phase (second liquid phase). This mixture is injected at 10 ml/min via a capillary. RESOMER? RP d 155 is a mPEG-PLA Diblock copolymers containing a 5 kDa mPEG block and 15 wt % PEG.

(80) The results are shown in table 11.

Example XII

(81) 180 mL of deionized water were initially charged in the reservoir of the apparatus and circulated at 3 000 rpm (first liquid phase). 16 mL of a stock solution consisting of 5.7% RESOMER? RP d 255, 51.3% ethanol, 43.0% DMSO served as organic phase (second liquid phase). This mixture is injected at 10 ml/min via a capillary. RESOMER? RP d 255 is a mPEG-PLA Diblock copolymers containing a kDa mPEG block and 25 wt % PEG.

(82) The results are shown in table 12.

Example XIII

(83) 180 mL of deionized water were initially charged in the reservoir of the apparatus and circulated at 3 000 rpm (first liquid phase). 16 mL of a stock solution consisting of 7% RESOMER? RP d 505, 62.6% ethanol, 30.4% DMSO served as organic phase (second liquid phase). This mixture is injected at 10 ml/min via a capillary. RESOMER? RP d 505 is a mPEG-PLA Diblock copolymers containing a 5 kDa mPEG block and 50 wt % PEG

(84) The results are shown in table 13.

Example Series 7: Biodegradable Microparticles as Pharmaceutical Carrier Systems for Pharmaceutical Applications

(85) As shown in example series 3 the levitronic mixing allows PLGA-based nanoparticles to be produced over a wide concentration range with a robust process with or without active ingredient loading. By process parameter- and excipient modification the herein described process can also be used to generate polymeric microparticles. There are examples for successfully marketed polymeric microparticles such as Bydureon? (AstraZeneca). Particles in the size range between 20 ?m and 90 ?m can be achieved by the herein described process. Such particles are considered as microcarrier.

(86) RESOMER? RG 502 H, a poly(D,L-lactide-co-glycolide), obtainable from Evonik, was employed as an example formulation. Deionized water was initially charged as first liquid phase and circulated at varying rates (pump speeds) and the organic PLGA solution was metered in at varying rates as a second liquid phase (dispersed phase/DP). To this end, RG 502 H was first completely dissolved in ethyl acetate with stirring. The concentration of the PLGA in the organic phase was varied between and 30%. As a third liquid phase (continuous phase/CP) a 2% PVA solution was added via the second capillary. The CP/DP ratio was varied between 1:1 and 1:5.

(87) In addition to placebo formulations, the PLGA microparticles were also successfully loaded with a model active ingredient, namely Meloxicam which is approved as a non-steroidal anti-inflammatory drug. In the verum experiments Meloxicam was dissolved at 5% in DMSO and added to the organic phase at 10% and 20% based on the PLGA mass.

(88) The resulting particles were filtered through a 20 ?m sieve and resuspended in water. The microparticles are suitable in terms of size and have a narrow particle size span which is indicative of quality. The obtained meloxicam microparticles have an encapsulation efficiency between 92.7% (10% Drugload) and 96.6% (20% Drugload). Deviating to all other examples, size determination of polymeric microparticles was conducted at a MasterSizer system (Malvern Panalytical)

(89) The results are shown in table 14.

(90) TABLE-US-00002 TABLE 1 Example I results Total lipids Sampling time concentration Total Pump Addition [min] (after Experiment (EPC-COL) volume in speed rate completed Zav number in EtOH system [ml] [rpm] [ml/min] addition) (backscattering) PDI 21836/21 200 mg/ml 777.5 ml (700 ml 14000 10 ml/min 10 153.2 0.309 ammonium 40 150.0 0.254 sulfate + 77.5 ml 70 147.5 0.246 EPC-col in EtOH) 100 144.3 0.253 21836/29_2 200 mg/ml 183.2 ml (150 ml 14000 5 ml/min 5 155.6 0.216 ammonium 30 156.2 0.219 sulfate + 33.2 ml 156.8 0.233 EPC-col in EtOH) 156.0 0.239

(91) TABLE-US-00003 TABLE 2 Example II results Total lipids Sampling time concentration Total Pump [min] (after Experiment (EPC-COL) volume in speed completed Zav number in EtOH system [ml] [rpm] addition) (backscattering) PDI 21836/20 400 mg/ml 500 ml (450 ml 10000 10 549.9 0.705 ammonium sulfate + 90 512.4 0.708 50 ml EPC-col in EtOH) 21836/24 400 mg/ml 777.5 ml (700 ml 14000 30 422.4 0.710 ammonium sulfate + 398.5 0.785 77.5 ml EPC-col in EtOH) 21836/26 400 mg/ml 777.5 ml (700 ml 8200 30 1001 0.582 ammonium sulfate + 973.1 0.838 77.5 ml EPC-col in EtOH)

(92) TABLE-US-00004 TABLE 3 Example III results Total lipids Addition rate Capillary concentration Ratio aqueous Total Pump of organic type/ Experiment (EPC-COL) in phase:organic volume in speed solution distance Zav number EtOH phase system [ml] [rpm] [ml/min] to pump (backscattering) PDI 21836/33_1 15.2 mg/ml 3:1 160 ml [120 ml 14000 10 ml/min Pipe ext 108.4 0.136 aqueous phase/40 diameter 106.5 0.097 ml organic phase] 3.25 mm/20 mm 21836/33_2 15.2 mg/ml 6:1 141 ml [120 ml 14000 10 ml/min Pipe ext 92.67 0.144 aqueous phase/21 diameter 91.53 0.113 ml organic phase] 3.25 mm/20 mm 21836/33_3 15.2 mg/ml 9:1 150 ml [135 ml 14000 10 ml/min Pipe ext 92.81 0.129 aqueous phase/15 diameter 88.88 0.159 ml organic phase] 3.25 mm/20 mm 21836/33_4 15.2 mg/ml 9:1 150 ml [135 ml 5000 10 ml/min Pipe ext 90.75 0.134 aqueous phase/15 diameter 89.37 0.110 ml organic phase] 3.25 mm/20 mm 21836/33_5 15.2 mg/ml 9:1 150 ml [135 ml 14000 10 ml/min Fine 81.12 0.160 aqueous phase/15 capillary/ 80.78 0.175 ml organic phase] 20 mm 21836/33_6 15.2 mg/ml 9:1 150 ml [135 ml 14000 5 ml/min Fine 81.56 0.202 aqueous phase/15 capillary/ 79.85 0.188 ml organic phase] 20 mm

(93) TABLE-US-00005 TABLE 4 Example IV results Total lipids Addition rate concentration Total Pump of organic Capillary Experiment (EPC-COL) volume in speed solution distance Zav number in EtOH system [g] [rpm] [ml/min] to pump (backscattering) PDI 21836/30_1 386.90 mg/ml 795 g [582.56 g 14000 5 ml/min 20 mm 238.3 0.216 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_2 386.90 mg/ml 795 g [582.56 g 14000 15 ml/min 20 mm 235.5 0.146 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_3 386.90 mg/ml 795 g [582.56 g 14000 25 ml/min 20 mm 224.7 0.173 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_4 386.90 mg/ml 795 g [582.56 g 14000 35 ml/min 20 mm 217.6 0.157 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_5 386.90 mg/ml 795 g [582.56 g 5000 15 ml/min 20 mm 314.4 0.286 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_6 386.90 mg/ml 795 g [582.56 g 10000 15 ml/min 20 mm 267.7 0.156 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_7 386.90 mg/ml 795 g [582.56 g 14000 15 ml/min 20 mm 234.9 0.125 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_8 386.90 mg/ml 795 g [582.56 g 14000 + 90 min 15 ml/min 20 mm 216.3 0.156 [38.69%] aqueous phase/ recirculation after 212.24 g organic completed addition phase] 21836/30_9 386.90 mg/ml 795 g [582.56 g 14000 15 ml/min 10 mm 238.6 0.232 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_10 386.90 mg/ml 795 g [582.56 g 14000 15 ml/min 20 mm 237.7 0.138 [38.69%] aqueous phase/ 212.24 g organic phase] 21836/30_11 386.90 mg/ml 795 g [582.56 g 14000 15 ml/min 30 mm 230.7 0.174 [38.69%] aqueous phase/ 212.24 g organic phase]

(94) TABLE-US-00006 TABLE 5 Example V results Total lipids Addition rate Capillary concentration Total Pump of organic distance Experiment (EPC-COL) in volume in speed solution to Zav number EtOH system [g] [rpm] [ml/min] pump (backscattering) PDI 21836/30_12 193.45 mg/ml 795 g [582.56 g 14000 15 ml/min 20 mm 197.9 0.126 [19.35%] aqueous phase/ 212.24 g organic phase]

(95) TABLE-US-00007 TABLE 6 Example VI results Pump Rate of Experiment Vessel/ speed addition Zav number Organic phase batch size Capillary [rpm] [ml/min] (backscattering) PDI 21836/30 ACN/10 mg/ml/ Steel tank/777.8 Wide 14000 10 ml/min 101.3 0.200 77.8 ml ml after addition of (3.25 mm ext 100.2 0.197 org. phase diameter) 21836/31_1 ACN/10 mg/ml/ Plexiglas tank/ fine 14000 5 ml/min 117.6 0.199 15 ml 150 ml after 116.6 0.202 addition of org. phase 21836/31_2 ACN/10 mg/ml/ Plexiglas tank/ Wide 10000 10 ml/min 88.43 0.213 15 ml 150 ml after (3.25 mm ext 87.77 0.202 addition of org. diameter) phase 21836/31_3 ACN/10 mg/ml/ Plexiglas tank/ Wide 5000 10 ml/min 106.0 0.164 15 ml 150 ml after (3.25 mm ext 105.2 0.170 addition of org. diameter) phase 21836/31_4 ACN/10 mg/ml/ Plexiglas tank/ Wide 2500 10 ml/min 99.70 0.214 15 ml 150 ml after (3.25 mm ext 98.41 0.204 addition of org. diameter) phase 21836/32_1 ACN/10 mg/ml/ Plexiglas tank/ Wide 7500 10 ml/min 92.83 0.194 15 ml 150 ml after (3.25 mm ext 91.79 0.160 addition of org. diameter) phase 21836/32_2 ACN/10 mg/ml/ Plexiglas tank/ Wide 10000 Bolus 102.6 0.205 15 ml 150 ml after (3.25 mm ext (15 ml 102.1 0.197 addition of org. diameter) in 15 sec) phase 21836/32_3 ACN/10 mg/ml/ Plexiglas tank/ Wide 10000 Slow (2.5 105.4 0.170 15 ml 150 ml after (3.25 mm ext ml/min) 103.7 0.177 addition of org. diameter) phase 1836/34_1 ACN/10 mg/ml/ Plexiglas tank/ Wide 10000 10 ml/min 98.06 0.251 15 ml + 1% 150 ml after (3.25 mm ext 96.94 0.239 Ritonavir addition of org. diameter) phase 1836/34_1 ACN/10 mg/ml/ Plexiglas tank/ Wide 10000 10 ml/min 102.0 0.272 15 ml + 2% 150 ml after (3.25 mm ext 98.06 0.199 Ritonavir addition of org. diameter) phase

(96) TABLE-US-00008 TABLE 7 Example VII results Total lipids Total Pump Addition rate of Capillary Experiment concentration volume in speed organic solution type/distance Zav number in EtOH system [g] [rpm] [ml/min] to pump (backscattering) PDI 21836/36_1 15 mM 150 ml 10000 15 ml/min for Dual capillary 5 min after (thereof 70 ml phosphate acetate buffer with (20 mm over addition: 7.5 mM buffer pH 7.4 Poly A via syringe levitronic 104.6 0.207 DODMA) initially as outer pump pump head) 101.9 0.216 phase in levitronic 5 ml/min for 30 min after pump. organic lipid phase addition: 60 ml acetate via peristraltic 97.86 0.197 buffer containing pump 95.91 0.200 17.4 mg Poly A added via capillary 20 ml lipid solution added via capillary

(97) TABLE-US-00009 TABLE 8 Example VIII results Total lipids Total Pump Addition rate of Capillary Experiment concentration in volume in speed organic solution type/distance Zav number EtOH system [g] [rpm] [ml/min] to pump (backscattering) PDI 21836/37 15 mM (thereof 150 ml 14000 5 ml/min for single 5 min after 7.5 mM 112.5 ml actetate organic lipid phase capillary addition: DODMA) buffer pH 4.0 via syringe pump (20 mm over 122.2 0.367 initially as outer levitronic 115.6 0.340 phase in levitronic pump head) 30 min after pump. addition: 37.5 ml lipid 135.4 0.292 solution added via 135.0 0.289 capillary After adjustment to pH 7: 115.1 0.258 112.7 0.244

(98) TABLE-US-00010 TABLE 9 Example IX results Total volume Pump Addition rate of Capillary Experiment N:P in system speed organic solution type/distance Zav number Verh?ltnis [g] [rpm] [ml/min] to pump (backscattering) PDI 21836/38 15 150 ml 10000 5 ml/min for Dual capillary 5 min after 80 ml of RNase- phases via 20 mm above metered free water as first capillary pump head addition: phase in pump 46.06 0.434 circuit 47.25 0.403 35 ml of aqueous 30 min after poly A solution metered via capillary addition: 35 ml of PEI 60.41 0.396 solution in 20 mM 58.79 0.422 acetate buffer via capillary

(99) TABLE-US-00011 TABLE 10 Example X results Total volume Pump Addition rate of Capillary Experiment N:P in system speed organic solution type/distance Zav number ratio [g] [rpm] [ml/min] to pump (backscattering) PDI 21836/39 15 150 ml 10000 5 ml/min Single 5 min after 115 ml of aqueous poly capillary metered A as first phase via (20 mm above addition: circuit pump head) 41.51 0.336 35 ml of PEI solution in 43.27 0.318 20 mM acetate buffer 30 min after via capillary metered addition: 102.4 0.224 59.99 0.325

(100) TABLE-US-00012 TABLE 11 Example XI results Experiment Vessel/ Pump speed Rate of addition Zav [nm] number Organic phase batch size Capillary [rpm] [ml/min] (backscattering) PDI E22RDI001175 16 ml of 6.2% Plexiglas tank/ Fine (1 mm inner 3000 10 ml/min 127.83 0.209 RESOMER? RP d 216 ml after diameter) 155, 55.8% ethanol, addition of org. 38.0% DMSO phase

(101) TABLE-US-00013 TABLE 12 Example XII results Experiment Pump speed Rate of addition Zav [nm] number Organic phase Vessel/batch size Capillary [rpm] [ml/min] (backscattering) PDI E22RDI001178 16 ml of 5.7% Plexiglas tank/ Fine (1 mm inner 3000 10 ml/min 53.62 0.108 RESOMER? RP d 216 ml after diameter) 255, 51.3% ethanol, addition of org. 43.0% DMSO phase

(102) TABLE-US-00014 TABLE 13 Example XIII results Experiment Pump speed Rate of addition Zav [nm] number Organic phase Vessel/batch size Capillary [rpm] [ml/min] (backscattering) PDI E22RDI001182 16 ml of 7% Plexiglas tank/ Fine (1 mm inner 3000 10 ml/min 34.94 0.080 RESOMER? RP d 216 ml after diameter) 505, 62.6% ethanol, addition of org. 30.4% DMSO phase

(103) TABLE-US-00015 TABLE 14 Example XIV results DP/CP Pump ration/Rate Particle Experiment speed of addition size number Organic phase Vessel/batch size Capillary [rpm] [ml/min] [?m] Span E21RDI006440 5% PLGA in Plexiglas tank/ Fine 3000 1:3/ 27.6 1.362 Ethylacetate 200 ml after (1 mm inner 2 ml/min (DP) (7 ml) addition of org. diameter) 6 ml/min (CP) phase E21RDI006441 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 39 1.363 Ethylacetate 200 ml after (1 mm inner 2 ml/min (DP) (7 ml) addition of org. diameter) 6 ml/min (CP) phase E21RDI006441 20% PLGA in Plexiglas tank/ Fine 3000 1:3/ 55.2 1.199 Ethylacetate 200 ml after (1 mm inner 2 ml/min (DP) (7 ml) addition of org. diameter) 6 ml/min (CP) phase E21RDI006442 5% PLGA in Plexiglas tank/ Fine 2000 1:3/ 38.4 1.233 Ethylacetate 200 ml after (1 mm inner 2 ml/min (DP) (7 ml) addition of org. diameter) 6 ml/min (CP) phase E21RDI006442 5% PLGA in Plexiglas tank/ Fine 4000 1:3/ 19.9 1.634 Ethylacetate 200 ml after (1 mm inner 2 ml/min (DP) (7 ml) addition of org. diameter) 6 ml/min (CP) phase E21RDI006443 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 35.5 1.314 Ethylacetate 200 ml after (1 mm inner 1 ml/min (DP) (7 ml) addition of org. diameter) 3 ml/min (CP) phase E21RDI006443 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 33.3 1.225 Ethylacetate 200 ml after (1 mm inner 4 ml/min (DP) addition of org. diameter) 12 ml/min phase (CP) E21RDI006443 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 34.4 1.239 Ethylacetate 200 ml after (1 mm inner 8 ml/min (DP) addition of org. diameter) 24 ml/min phase (CP) E21RDI006443 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 29.4 1.561 Ethylacetate 200 ml after (1 mm inner 16 ml/min addition of org. diameter) (DP) 48 phase ml/min (CP) E21RDI006443 10% PLGA in Plexiglas tank/ Fine 3000 1:1/ 36.5 1.328 Ethylacetate 200 ml after (1 mm inner 8 ml/min (DP) addition of org. diameter) 8 ml/min (CP) phase E21RDI006443 10% PLGA in Plexiglas tank/ Fine 3000 1:5/ 31.0 1.494 Ethylacetate 200 ml after (1 mm inner 8 ml/min (DP) addition of org. diameter) 40 ml/min phase (CP) E21RDI006460 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 45.2 1.509 Ethylacetate (7 ml) 200 ml after (1 mm inner 8 ml/min (DP) 10% Meloxicam in addition of org. diameter) 24 ml/min relation to polymer phase (CP) E21RDI006465 10% PLGA in Plexiglas tank/ Fine 3000 1:3/ 87.6 1.199 Ethylacetate (7 ml) 200 ml after (1 mm inner 8 ml/min (DP) 20% Meloxicam in addition of org. diameter) 24 ml/min relation to polymer phase (CP)

LIST OF REFERENCE NUMERALS

(104) 0 Apparatus 1 First reservoir vessel 2 Second reservoir vessel 3 Active element 4 First feed conduit Second feed conduit 6 Pump 7 Propulsion means 7+ Rotating field 8 Discharge 8+ Circuit 9 Withdrawal fitting 11 Frame 11 Cone 12 First pipeline 13 Ground end 14 Second pipeline 15 Third pipeline 16 Inner wall 17 Housing 18 Rotor 111 Insert 1111 Charging fitting