Lecithin particles in the nanometer range as a carrier of the active ingredient for parenteral administration

Description

[0001] Parenterals are sterile pharmaceutical preparations intended for administration into human or animal tissue by injection, infusion, or implantation. Such products must be free of large solids.

[0002] The parenteral administration of lipophilic, poorly water-soluble active substances is difficult because these substances usually do not dissolve in water in the necessary concentration and parenterals of poorly water-soluble active substances would therefore not be free of solids. Then, excipients must be added as solubilizers to improve the aqueous solubility of the active ingredient, for example organic solvents or surfactants or co-solvents, such as Tween 80, Tween 20, poloxamer, propylene glycol, glycerol, ethanol, polyethylene glycol, sorbitol, dimethylacetamide, Cremophor EL (Pramanick, S., Singodia, D., & Chandel, V. (2013). Excipient Selection In Parenteral Formulation Development. Pharma Times, 45(3), 65-77; Bittner, B. & Mountfield, RJ, Intravenous administration of poorly soluble new drug entities in early drug discovery: the potential impact of formulation on pharmacokinetic parameters. Curr. Opinion Drug Discov. Dev. 5, 59-71 (2002)).Cremophor EL, in particular, can cause anaphylactic reactions after parenteral administration (Theis, JG et al. Anaphylactoid reactions in children receiving high-dose intravenous cyclosporine for reversal of tumor resistance: the causative role of improper dissolution of cremophor EL. J. Clin. Oncol. 13, 2508-2516 (1995); O'Dwyer, PJ & Weiss, RB Hypersensitivity reactions induced by etoposide. Cancer Res. 68, 959-961 (1984)). Co-solvents - e.g., alcohols - are undesirable, especially when parenteral products are used in children.

[0003] Other excipients may cause pain during injection (Duma, RJ, Akers, MJ & Turco, SJ in Pharmaceutical Dosage Forms: Parenteral Medications 2nd edn Vol. 1 (eds Avis, KE, Lieberman, HA & Lachman, L.) 17-58 (Marcel Dekker, New York, 1992)) or precipitation of the active ingredient in the blood after dilution of the injected solution. Precipitation of the active ingredient after application can result in local damage to the vessel wall due to the often sharp-edged active ingredient crystals (phlebitis) (Ward, GH & Yalkowsky, SH Studies in Phlebitis. VI. Dilution-induced precipitation of amiodarone HCl. J. Parenter. Sci. Technol. 47, 161-165 (1993). Davio, SR et al. Precipitation of the renin inhibitor ditekiren upon iv infusion; in vitro studies and their relationship to in vivo precipitation in the cynomolgus monkey. Pharm. Res. 8, 80-83 (1991)).

[0004] Cyclodextrins are also used as solubilizers for lipophilic substances. These are themselves highly soluble in water and have the ability to encapsulate nonpolar active ingredients. This allows for solid-free pharmaceutical formulations in which lipophilic active ingredients are complexed in larger concentrations (e.g., the active ingredient carfilzomib in Kyprolis™). The high cost of these special excipients is a disadvantage.

[0005] By converting the poorly water-soluble active ingredient into nanoparticles, problematic excipients in high concentrations can be dispensed with and their side effects can be prevented.

[0006] In addition, the infusion volume for administering the same dose can be reduced because the water solubility of the active ingredient can be exceeded, thus generating higher drug concentrations in aqueous medium.

[0007] When chemical stability in aqueous media is limited, encapsulation in nanoparticles very effectively protects the active ingredient from undesirable chemical reactions. This can increase product stability and, for example, enable product storage at room temperature instead of refrigeration. Furthermore, it can enable storage of the pharmaceutical formulation as a liquid suspension, eliminating the need for conversion into a powder for reconstitution (ready-to-use, "RTU"). A chemically labile active ingredient does not need to be formulated as a powder, which must be resuspended or dissolved before administration to ensure chemical stability during storage; in this case, the active ingredient can be administered as a finished medicinal product. This avoids the general risk of medication errors due to handling errors during dilution or reconstitution and dilution.The main potential clinical advantages of converting the poorly water-soluble drug into nanoparticles are: . 1. Reduction in the required amount of solubilising surfactants and thus reduction in toxicity from solubilisers such as Cremophor EL; 2. No need for solubilising co-solvents such as organic solvents; 3. Reduction in injection volumes as the concentration of lipophilic active ingredients in the aqueous medium can be increased (beyond saturation solubility); 4. The possibility of altering the pharmacokinetics and dynamics of the active ingredient or drug: higher doses may be better tolerated due to a reduction in side effects, lower doses lead to comparable effectiveness, treatment cycles can be adapted; ( Wong, J., Brugger, A., Khare, A., Chaubal, M., Papadopoulos, P., Rabinow, B. E., Ning, J. (2008). Suspensions for intravenous (IV) injection: a review of development, preclinical and clinical aspects. Advanced Drug Delivery Reviews, 60(8), 939-954 ).

[0008] The size of the generated particles significantly controls their properties and biological behavior and is therefore a critical quality feature. The literature shows that, particularly in solid tumors, the permeability of blood vessels in the tumor can and does lead to an accumulation of active substances administered as nanoparticles (Fang, J.; Nakamura, H.; Maeda, H. The EPR Effect: Unique Features of Tumor Blood Vessels for Drug Delivery, Factors Involved, and Limitations and Augmentation of the Effect. Adv. Drug Delivery Rev. 2011, 63, 136-151; Rabinow, BE, Nanosuspensions in drug delivery. Nature Reviews. Drug Discovery, 3(9), 785-796 (2004)). This phenomenon is referred to as passive drug targeting. This increases the efficacy of therapy and, under certain circumstances, allows the use of lower drug doses. The optimal particle size is described as 50 to 200 nm.

[0009] It is therefore important to be able to adjust the particle size, as this is what enables passive drug transport and a particle size of less than 200 nm is necessary to enable filtration with a pore size of 0.22 µm as a terminal sterilization process, especially for heat-sensitive active ingredients and excipients that cannot be sterilized by thermal methods such as autoclaving.

[0010] The rate of release of the active ingredient from a particulate formulation into the blood (or tissue) determines, among other things, the drug's pharmacokinetics and dynamics. In addition to the excipient composition, the particle size also plays a role in determining the rate of release of the active ingredient. As the particle size decreases, the surface area increases. This leads to a more rapid dissolution of the particles and thus a more rapid release of the active ingredient.

[0011] To reduce the particle size of pharmaceutical active ingredients, EP 2 395 978 B1 proposes so-called solvent / non-solvent precipitation. Solvent / non-solvent precipitation means that a substance dissolved in a solvent collides head-on as a liquid stream with a second liquid stream, precipitating the dissolved substance. This so-called jet impingement creates a highly turbulent mixing zone of solvent and non-solvent and is suitable for the precipitation of particles in the nanometer range. EP 2 395 978 B1 describes solvent / non-solvent precipitation in the presence of surface-active molecules using a jet impingement reactor, in this case a microjet reactor according to EP 1 165 224 B1.Such a reactor has at least two opposing nozzles, each with an associated pump and supply line for spraying two liquid media. The liquid streams collide head-on at a common collision point in a reactor chamber. An opening is provided to guide the resulting suspension out of the reactor chamber.

[0012] WO 2016 / 146516 A1 discloses a method for producing nanoparticles. It involves providing a solvent stream containing an organic solvent, a glycerophospholipid (preferably phosphatidylcholine), and an active ingredient. After head-on collision with an aqueous stream at high pressure, nanoparticles with a size of 170 nm are produced.

[0013] The development of nanoparticulate formulations for parenteral administration is specifically tailored to the active ingredient, indication, dose, and route of administration. This development is time-consuming and costly. Therefore, there is a need for an improved process for the production of carrier nanoparticles into which a poorly water-soluble active pharmaceutical ingredient (API) can be embedded. This process also allows for easy adjustment of the particle size, particularly enabling the production of particles smaller than 0.22 µm.

[0014] This object is achieved by a method according to claim 1. Accordingly, the invention provides a method for producing a carrier system, wherein the carrier system contains a pharmaceutical active ingredient and at least one glycerophospholipid and wherein the method comprises the steps: Providing a solvent stream containing an organic solvent containing at least one glycerophospholipid and the pharmaceutical active ingredient and possibly a surface-active agent (surfactant), providing an aqueous stream containing possibly a surface-active agent (surfactant), and mixing the solvent stream and the aqueous stream by frontal collision, whereby the carrier system is precipitated as nanoparticles, characterized in that the average particle size of the precipitated nanoparticles measured by dynamic light scattering (DLS) at a temperature of 25 °C is 10 to 500 nm, the solvent stream contains 1 to 1000 mg / mL of at least one glycerophospholipid and the hydrodynamic pressure in the aqueous stream is at least 1 bar amounts.

[0015] The inventors have invented a time- and cost-saving process in which glycerophospholipids can be used as the main components of a carrier system for active ingredients (especially water-insoluble ones). Glycerophospholipids are safe even when administered parenterally and can be used in high concentrations. Even water-insoluble active ingredients can be formulated using this process in concentrations above their aqueous saturation solubility. Furthermore, the process makes it possible to produce a desired particle size by increasing the hydrodynamic pressure. This enables sterile filtration and passive drug transport. By embedding them in a lipid matrix, the process according to the invention can protect chemically labile active ingredients from degradation. A particular advantage is that the process can be used as a standard manufacturing process for a wide variety of active ingredients.By selecting the particle size and other components of the carrier system, the release of the active ingredient from the carrier system can also be modified. The release of the active ingredient can be accelerated by reducing the size of the particles. The particle size can be adjusted via hydrodynamic pressure. The incorporation of active ingredient leads to only a slight increase in particle size; the relationship between increasing pressure and decreasing particle size remains intact and significantly influences the particle size.

[0016] By mixing the solvent stream and the aqueous stream, a suspension is obtained in which the carrier system is precipitated as nanoparticles. The carrier system is a solid containing a pharmaceutically active ingredient, at least one glycerophospholipid, and optionally other ingredients, e.g., cholesterol or one of its derivatives, and is present in the form of nanoparticles. The surfactants are not part of the solid particle, but are present in a dissolved state in the suspension or in a wetting state on the surface of the particle.

[0017] The pharmaceutical active ingredient is, in particular, a low-molecular-weight compound (a so-called "small molecule") or a peptide. However, the term "pharmaceutical active ingredient" also encompasses antibodies, RNA, proteins, etc. In one embodiment, the pharmaceutical active ingredient is water-insoluble. A water-insoluble pharmaceutical active ingredient has a solubility of < 1 mg / ml ("very slightly soluble") and preferably < 0.1 mg / ml ("practically insoluble") in water. The process is suitable for water-insoluble active ingredients. The term "at least one glycerophospholipid" encompasses one or more glycerophospholipids. The term "glycerophospholipids" (phosphoglycerides, phosphoglycerolipids) includes phosphatidylcholines (lecithins), phosphatidylethanolamines, phosphatidylserines, and phosphatidylinositols.

[0018] Phosphatidylcholines (lecithins) are particularly preferred as the at least one glycerophospholipid. Phosphatidylcholine or lecithin (e.g., Lipoid S 100) is, for example, a suitable, commercially available glycerophospholipid.

[0019] The organic solvent can be one organic solvent or a mixture of several, e.g., two, organic solvents. Preferably, the organic solvent(s) is / are miscible with water. If multiple organic solvents are used, they are miscible with each other.

[0020] The main component (e.g., ≥ 50, 60, 70, 75, 80, 90, 95, or 99 wt%) of the aqueous stream is water. The aqueous stream consists of water and optionally contains excipients such as surfactants, buffers, etc.

[0021] The term nanoparticle refers to a solid in the form of particles with an average particle size between 1 and 1000 nm.

[0022] Mixing the solvent stream and the aqueous stream involves colliding the streams. Preferably, the streams are arranged at an angle of approximately 180° to each other. In other words, the streams meet head-on.

[0023] According to the invention, the average particle size can be measured by dynamic light scattering (DLS) (e.g., with a Malvern Zetasizer AT / Advance Range / ZS90 / ZS or Malvern Ultra from Malvern Instruments Ltd.; Nanophox and other DLS measuring devices from Wyatt Technology Corporation; Litesizers from Anton Paar GmbH) at a temperature of 25°C. This temperature is kept constant during the measurement. The z-average diameter is calculated together with the polydispersity index (PDI), preferably from the cumulant analysis of the autocorrelation function of the measured DLS intensity, as defined in ISO 22412:2008. Particle sizes are preferably measured with a Zetasizer from Malvern Instruments Ltd.

[0024] The PDI is a dimensionless estimate of the width of the particle size distribution, scaled from 0 to 1. The term average particle size refers to the z-average diameter.

[0025] In general, hydrodynamic pressure can be calculated either theoretically or monitored using a pressure gauge. Hydrodynamic pressure is calculated using the following formula: p = 1 2 ∗ ρ ∗ Q π ∗ r 2 2 ∗ 10 − 5 where p = the pressure in the stream ([bar]), ρ = the density of the composition (all components) of the stream ([kg / m 3< ]), Q = the flow rate of the stream ([m 3< / s]) and r = the radius of the nozzle ([m]).

[0026] The speed of the current v (m / s) can be calculated using the following formula: v = Q π ∗ r 2 2

[0027] The flow rate is one of the variables for pressure calculation. It is preferable to ensure that the pumps that drive the flow are functional and meet the required pumping capacity before starting the production process. The flow rate can be verified—without using the reactor—by pump calibration: A defined flow rate is set for each pump, pumping is carried out for a certain time, and the pumped volume is checked for weight using a scale. Experimental and theoretical weights are compared.

[0028] Inline flow rate monitoring can be achieved by implementing flow rate meters in the reactor feed lines, allowing flow rates to be monitored during the process. Bronkhorst CORI FLOW meters or Coriolis flow meters can be used for this purpose.

[0029] An essential feature of the invention is the hydrodynamic pressure, or the velocity of the colliding streams. As explained above, the hydrodynamic pressure is calculated from the flow rate and density of the streams (jets), as well as from the radius of the nozzles through which the streams are ejected into the reactor chamber (chamber in which the two streams are mixed). To achieve a hydrodynamic pressure according to the invention, it is generally preferred that the flow rate of the solvent stream is 1 ml / min to 10 l / min, more preferably 10 ml / min to 1.33 l / min, most preferably 50 ml / min to 250 ml / min, and the flow rate of the aqueous stream is 1 ml / min to 10 l / min, more preferably 10 ml / min to 1.33 l / min, most preferably 50 ml / min to 250 ml / min.It is preferred that the velocity of the solvent stream is 2 to 150 m / s, preferably 10 to 125 m / s, and the velocity of the aqueous stream is 2 to 150 m / s, preferably 20 to 125 m / s.

[0030] The density of the solvent stream is preferably 0.600 to 1.300 g / mL, and the density of the aqueous stream is preferably 0.600 to 1.300 g / mL. The diameter (radius = half the diameter) of the nozzles should preferably be 5 to 5,000 µm, preferably 10 to 3,000 µm, more preferably 10 to 2,000 µm, and most preferably 50 to 500 µm.

[0031] Preferably, the distance between the opposite nozzles (for the two streams) is less than 5 cm, preferably less than 3 cm and preferably less than 1 cm.

[0032] Thus, depending on the choice of nozzle diameter and the selected mixing ratio, hydrodynamic pressures for the solvent stream are >0.02 bar, preferably >0.5 bar, more preferably >10 bar, and most preferably >30 bar. Depending on the choice of nozzle diameter, hydrodynamic pressures for the aqueous stream are >0.02 bar, preferably >0.5 bar, more preferably >10 bar, and most preferably >30 bar. Preferably, the hydrodynamic pressure in the solvent stream is equal to or less than the hydrodynamic pressure of the aqueous stream.

[0033] The reactor chamber (the chamber in which the two streams collide and are mixed) can be pressurized with gas. Gas, particularly inert gas or inert gas mixtures, but also reactive gas, can be introduced into the reactor chamber through a gas inlet. The process is preferably carried out without gas. The gas pressure in the reactor chamber should preferably be 0.05 to 30 bar, preferably 0.2 to 10 bar, and particularly preferably 0.5 to 5 bar.

[0034] Preferably, the process is a continuous process.

[0035] Preferably, the suspension obtained by mixing or the carrier system contained therein is further processed, preferably into a pharmaceutical composition. This may include one or more steps, for example, removal of the organic solvent, adjustment of the pH, adjustment of the buffer capacity, addition of antioxidants, adjustment of the osmolality, terminal sterilization by sterile filtration, freeze-drying, filling, etc.

[0036] In one embodiment, the suspension obtained by mixing or a suspension containing the carrier system is subjected to sterile filtration. The filter material is selected, for example, from cellulose acetate, regenerated cellulose, polyamide, polyethersulfone (PES), modified polyethersulfone (mPES), polyvinylidene fluoride (PVDF), and polytetrafluoroethylene (PTFE). Sterile filtration enables parenteral administration of the product and, in accordance with legal requirements, is the method of choice for parenteral products that cannot be terminally sterilized by moist or dry heat. Thanks to the process according to the invention, the particle size can be precisely adjusted. This is important because the loss of solids during filtration can be minimized if the nanoparticles have an average particle size below 210 nm (e.g., 150 nm) and a low PDI (when choosing a suitable filter material).

[0037] The verbs "comprising" and "containing" and their conjugations encompass the concept "consist of" and its conjugations. The terms "a" or "an" encompass the concept "at least one."

[0038] Embodiments of the invention may be combined in any way, unless this is clearly not intended. Preferred embodiments of the invention are also specified in the dependent claims. Preferably, the average particle size of the precipitated nanoparticles is 25 to 500 nm, more preferably 50 to 250 nm, or 70 to 170 nm, or 80 to 150 nm, or 90 to 110 nm.

[0039] In particular, the above average particle sizes are achieved with the following hydrodynamic pressures: According to the invention, the hydrodynamic pressure in the aqueous stream is at least 1 bar, particularly preferably 2 bar to 60 bar.

[0040] Preferably, the hydrodynamic pressure in the solvent stream is at least 0.01 bar, preferably more than 0.1 bar, more preferably more than 1 bar, most preferably 2 bar to 60 bar.

[0041] Preferably, the average particle size of the precipitated nanoparticles is adjusted by the hydrodynamic pressure in the aqueous stream and / or the hydrodynamic pressure in the solvent stream, with higher hydrodynamic pressure precipitating nanoparticles of smaller average particle size. This applies particularly at a constant mixing ratio of aqueous stream to solvent stream. Preferably, the at least one glycerophospholipid is selected from phosphatidylcholines.

[0042] According to the invention, the solvent stream contains 1 to 1000 mg / mL, preferably 50 to 700 mg / mL, 100 to 500 mg / mL, 100 to 300 mg / mL, for example, about 150 or about 225 mg / mL, of the at least one glycerophospholipid. The content of the at least one glycerophospholipid is important for the formation of the carrier system, the stabilization, and the release of the active ingredient.

[0043] The solvent stream and / or the aqueous stream preferably contains 1 to 250 mg / mL, preferably 5 to 100 mg / mL, particularly preferably 8 to 50 mg / mL, of a steroid, preferably cholesterol or one of its derivatives. The content of cholesterol or one of its derivatives can support the formation of very small carrier systems (see, for example, the experimental section). It may also be preferable to adjust the pH of the aqueous stream to 5 to 2.5, especially if the cholesterol derivative is present in the aqueous stream. Deoxycholic acid and / or sodium deoxycholate are particularly preferred as cholesterol derivatives.

[0044] Preferably, mixing the solvent stream and the aqueous stream comprises mixing in a volume ratio of solvent stream to aqueous stream of between 1:1 to 1:20, more preferably between 1:1 to 1:10.

[0045] Optionally, the aqueous stream contains 0.1 to 250 mg / mL, preferably 0.5 to 100 mg / mL, particularly preferably 1.0 to 15 mg / mL, of one or more ionic or non-ionic surfactants, wherein the surfactant is preferably sodium oleate and / or oleic acid.

[0046] In one embodiment, the solvent stream and / or the aqueous stream further contains one or more surfactants selected from polyethoxylated castor oil (Kolliphor ELP), Macrogol 15 hydroxystearate (Kolliphor HS15), pegylated castor oil, docusate sodium, poly(ethylene glycol)- block -poly(propylene glycol)- block- Poly(ethylene glycol) (Poloxamer 188), vitamin E and derivatives thereof, polysorbate 20 (Tween 20), polysorbate 40 (Tween 40), polysorbate 60 (Tween 60), and polysorbate 80 (Tween 80). Surfactants can influence the formation of the carrier system, stabilize the carrier system, and influence the release of the active pharmaceutical ingredient.

[0047] Preferably, the organic solvent is a water-miscible solvent, wherein the organic solvent is preferably i) ethanol, or ii) a mixture of ethanol with another organic solvent that is miscible with water and ethanol, or iii) acetone. The ethanol can be pure ethanol or aqueous ethanol, i.e., ethanol with a specific water content (e.g., 85 vol%). Preferably, the other organic solvent is acetone.

[0048] In a particular embodiment, the aqueous stream contains 1 to 1000 mM (acid plus conjugate base), preferably 20 to 500 mM or 20 to 250 mM (acid plus conjugate base), of a buffer system.

[0049] Preferably, the concentration (mg / mL) of pharmaceutically active ingredient in the solvent stream is significantly lower than the concentration (mg / mL) of the glycerophospholipid. Since both components, active ingredient and carrier material (glycerophospholipid), coprecipitate, the resulting particles consist of significantly more carrier material than active ingredient. The properties of the resulting active ingredient-containing particles are therefore largely determined by the carrier material.

[0050] The solvent stream preferably contains 0.25 to 250 mg / mL, preferably 1 to 150 mg / mL, particularly preferably 10 to 100 mg / mL, for example, about 50 mg / mL, of the active pharmaceutical ingredient. The content of the active pharmaceutical ingredient in the solvent stream can be important for the formation of the carrier system, the stabilization, and the release of the active ingredient.

[0051] In one embodiment, a jet impingement reactor is used to carry out the process. An example of such a reactor is known from EP 1 165 224 B1. Other micromixing devices and / or precipitation devices with colliding streams can also be used, e.g., a device as described in WO 2018 / 234217 A1.

[0052] In one embodiment, the solvent stream and the carrier system contain hydroxypropyl methylcellulose acetate succinate (HPMCAS). In this embodiment, the organic solvent is preferably acetone. HPMCAS precipitates upon mixing with water and thus forms a component of the carrier system. In this embodiment, the concentration of HPMCAS in the solvent stream is preferably 1 to 100 mg / mL, more preferably 5 to 30 mg / mL, and the concentration of the pharmaceutically active ingredient in the solvent stream is preferably 1 to 30 mg / mL, more preferably 2 to 15 mg / mL. In this embodiment, the pH of the aqueous stream is preferably 6 to 8, more preferably 6 to 7.

[0053] The invention is illustrated below by non-limiting examples: Example 1: Production of drug-free carrier particles (not according to the invention)

[0054] The organic phase contained 225 mg / ml Lipoid S 100 in EtOH / acetone 1:1 (v / v). The aqueous phase contained 10.5 mg / ml sodium oleate. One portion of the organic phase was mixed with five portions of the aqueous phase in a MicroJet reactor with two nozzles of 200 µm each ( Fig. 1 , unfilled quadrilaterals) or 300 µm each ( Fig. 1 , unfilled points) mixed.

[0055] How Fig. 1 Depending on the selected hydrodynamic pressure, carrier particles between 150 nm and 79 nm are obtained. The higher the hydrodynamic pressure, the smaller the resulting particles.

[0056] The efficiency of the mixture is shown by the comparison with the manual precipitation by mixing the organic phase and the aqueous phase in a beaker ( Fig. 1 , unfilled triangle).

[0057] Placebo data are summarized in Table 1. Table 1. Nozzle diameter / µm Solvent flow rate / ml / min Solvent speed / m / s Solvent pressure / bar Flow rate e water / ml / min Water speed / m / s Water pressure / bar Particle size / nm PDI 245 0,26 300 30 7,1 0,20 150 35,4 6,3 89 0,22 300 20 4,7 0,09 100 23,6 2,8 105 0,22 300 10 2,4 0,02 50 11,8 0,7 150 0,26 200 40 21,2 1,80 200 106,1 56,3 - - 200 36 19,1 1,46 180 95,5 45,6 79 0,26 200 30 15,9 1,01 150 79,6 31,7 76 0,28 200 25,6 13,6 0,74 128 67,9 23,1 76 0,23 200 20 10,6 0,45 100 53,1 14,1 86 0,25 200 13,3 7,1 0,20 66,7 35,4 6,3 100 0,23 200 10 5,3 0,11 50 26,5 3,5 115 0,24 Example 2: Inventive production of active ingredient-containing particles (Formulation 1)

[0058] A lipophilic, water-insoluble active ingredient with a molecular weight of 400 g / mol was incorporated into carrier particles similar to those described in Example 1. The experiment was conducted as above using nozzles with a diameter of 300 µm, except that the organic phase additionally contained 50 mg / mL of the active ingredient. The aqueous phase contained 1 mg / mL of sodium oleate.

[0059] The incorporation of active ingredient leads to an increase in particle size of approximately 30 nm in direct comparison to Example 1. It should be noted that the composition of Formulation 1 and the placebo from Example 1 differs in the concentration of sodium oleate. Depending on the selected hydrodynamic pressure in the aqueous stream, particles between 140 nm and 78 nm were obtained ( Fig. 1 , filled dots). The higher the hydrodynamic pressure, the smaller the resulting particles.

[0060] One part of the organic phase was mixed with five parts of the aqueous phase in a MicroJet reactor with two nozzle orifices of 300 µm each. The data are summarized in Table 2. Table 2. Nozzle diameter / µm Solvent flow rate / ml / min Speed ​​of solvent / m / s Solvent pressure / bar Flow rate water / ml / min Water speed / m / s Pressure water / bar Particle size / nm P DI 300 50 11,8 0,56 250 58,9 17,3 78 0, 27 300 20 4,7 0,09 100 23,6 2,7 133 0, 10 300 20 4,7 0,09 100 23,6 2,7 121 0, 17 300 20 4,7 0,09 100 23,6 2,7 139 0, 20 Example 3: Inventive production of active ingredient-containing particles (Formulation 2)

[0061] The experiment was carried out as in Example 2, but the aqueous phase contained not 1 mg / ml sodium oleate, but 17 mg / ml sodium oleate.

[0062] Depending on the selected hydrodynamic pressure, particles between 154 nm and 88 nm are obtained. The higher the hydrodynamic pressure, the smaller the resulting particles. ( Fig. 1 , filled diamonds). Data are summarized in Table 3. Table 3. Nozzle diameter / µm Solvent flow rate / ml / min Solvent speed / m / s Solvent pressure / bar Flow rate water / ml / min Water speed / m / s Pressure water / bar Particle size / nm PDI 300 50 11,8 0,56 250 58,9 17,4 88 0,25 300 20 4,7 0,09 100 23,6 2,8 154 0,15 Example 4: Composition of the product

[0063] The concentrations of the starting solutions as well as the concentrations in the suspension for Examples 1 to 3 are summarized in Table 4: Table 4. Organic phase EtOH / acetone 1:1 (v / v) Aqueous phase Concentration in suspension in mg / ml API Lipoid 5100 NaOleat API Lipoid S100 NaOleat mg / ml mg / ml mg / ml mg / ml mg / ml mg / ml placebo 0 225 10,5 0 37,5 8,75 Formulation 1 50 225 1 8,3 37,5 0,8 Formulation 2 50 225 17 8,3 37,5 14,2

[0064] The concentration of the active ingredient is significantly lower than that of the carrier material, lecithin. The theoretical active ingredient content of the particles is 18.2%. The resulting particles therefore contain significantly more carrier material than active ingredient.

[0065] Example 4: Solvent removal using cross flow filtration CFF using formulation 2 from example 3

[0066] CFF experiments were performed using a 100 kDa PES hollow fiber from Repligen and five wash volumes. Water with 10 mg / mL or 5 mg / mL sodium oleate was used as wash medium.

[0067] Formulation 2 was prepared for these experiments with a flow rate of 50 ml / min for the organic phase and a nozzle diameter of 300 µm. Particle sizes below 90 nm were achieved. Solvent removal using CFF was possible without loss of active ingredient if the suspension was pre-diluted 1:1 prior to CFF and washed with 10 mg / ml sodium oleate. If no dilution step was included or a lower surfactant concentration was used, active ingredient losses occurred, as shown in Table 5. Table 5. ID [NaOleat] in CFF washing medium Pre-CFF Post-CFF PS nm [API] (mg / mL) PS nm [API] (mg / mL) 2 10 mg / mL 87,8 8,2 86,4 5,5 3 10 mg / mL 85,1 4,0 82,7 5,4 4 5 mg / mL 88,5 3,2 Example 5: Sterile filtration of the formulation from Example 4

[0068] ID 3 from Example 4 was tested for sterile filtration. High recovery values ​​were achieved with both PES and PVDF materials (see Table 6). Table 6. filter Active ingredient content before filtration Active ingredient content after filtration Recovery (mg / mL) (mg / mL) % CA 0.22 µm 5,65 5,17 91,50 PES 0.22 µm 5,58 98,76 PTFE 0.22 µm 5,34 94,51 PVDF 0.22 µm 5,57 98,58 Nylon 0.22 µm 5,17 91,50 Example 6: Stability data

[0069] Formulation 3 was subjected to an initial stability test at room temperature. The suspension demonstrated both physical and chemical stability over a 1-month observation period (see Table 7). Table 7. T=0 T=1M Particle size [nm] 106,9 106,5 PDI 0,29 0,28 pH 7,5 7,5 Content [mg / ml] 5,65 5,71 Example 7:

[0070] Further nanoparticles with a different active ingredient were produced according to the invention. The active ingredient was a peptide consisting of 20 amino acids and weighing 2180 Daltons. The peptide is water-soluble at 10 mg / mL at pH 3.5.

[0071] A solution containing 50 mg / mL of the active ingredient and 150 mg / mL of Lipoid S 100 was prepared in a mixture (solvent) of ethanol (85 vol%) and water (15 vol%). This solution was collided in a stream with an aqueous stream (non-solvent) containing various amounts of sodium deoxycholate and, if necessary, hydrochloric acid. Table 8 shows the conditions: Table 8. formulation Concentration in the solvent stream (85:15 EtOH:H2O) [mg / mL] Concentration in non-solvent [mg / mL] Solvent / Non-Solvent Ratio Active ingredient Lipoid S 100 Non-Solvent Sodium deoxycholate 359 50 150 HCl pH 3.5 25,0 0,5 385 HCl pH 3.5 10,0 0,5 414 HCl pH 3.0 0,5 0,5 415 HCl pH 3.0 1,0 0,5 416 HCl pH 3.0 2,0 0,5 417 HCl pH 3.0 5,0 0,5 422 HCl pH 3.0 7,5 0,5 418 Water 0,5 0,5 419 Water 1,0 0,5 420 Water 2,0 0,5 421 Water 5,0 0,5 423 Water 7,5 0,5

[0072] The average particle size and size distribution were analyzed and the results are shown in Table 9: Table 9. formulation average particle size [nm] PDI 359 211,7 0,180 385 214,8 0,200 414 438,4 0,140 415 487,8 0,273 416 230,5 0,067 417 557,1 0,224 422 750,3 0,385 418 723,4 0,319 419 496,9 0,271 420 537,0 0,273 421 549,9 0,306 423 605,2 0,508 Example 8:

[0073] Further nanoparticles containing the active ingredient as in Example 5 were prepared according to the invention. Table 10 shows the test conditions: Table 10. formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Concentration in non-solvent [mg / mL] Solvent / Non-Solvent Ratio Active ingredient Lipoid S 100 oleic acid Sodium oleate Non-Solvent Sodium oleate 424 50 150 2,0 - 25.0 mg / mL sodium deoxycholate in water - 0,5 425 5,0 - - 426 10,0 - - 427 20,0 - - 428 - 2,0 - 429 - 5,0 - 430 - - 5,0 431 - - 10,0

[0074] The average particle size and size distribution were analyzed and the results are shown in Table 11: Table 11. formulation average particle size [nm] PDI 424 209,2 0,041 425 115,6 0,131 426 260,0 0,154 427 548,2 1,000 428 255,5 0,200 429 546,9 1,000 430 324,0 0,613 431 338,6 0,012 Example 9:

[0075] Further nanoparticles containing the active ingredient as in Example 5 were prepared according to the invention. Table 12 shows the test conditions: Table 12. formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Concentration in non-solvent [mg / mL] Solvent / Non-Solvent Ratio Active ingredient Lipoid S100 oleic acid Sodium oleate Non-Solvent Sodium oleate 432 50 150 2,0 - Water - 0,5 433 5,0 - - 434 10,0 - - 435 20,0 - - 436 2,0 - - 437 - 5,0 - 438 - - 5,0 439 - - 10,0

[0076] The average particle size and size distribution were analyzed and the results are shown in Table 13: Table 13. formulation average particle size [nm] PDI 432 270,0 0,057 433 265,0 0,193 434 225,9 0,168 435 225,6 0,235 436 410,8 0,049 437 388,9 0,117 438 198,4 0,241 439 256,7 0,401 Example 10:

[0077] Further nanoparticles containing the active ingredient as in Example 5 were prepared according to the invention. Table 14 shows the test conditions: Table 14. Formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Non-solvent (mg / mL) Solvent / Non-Solvent Ratio Lipoid S100 Active ingredient Sodium oleate Tween 20 Tween 60 Tween 80 152 150 50 10 0,5 1:2 153 150 50 0 0,5 1:2 154 150 50 10 0,25 1:2 155 150 50 0 0,25 1:2 156 150 50 10 0,1 1:2 157 150 50 0 0,1 1:2 158 150 50 10 0,5 0,5:2,5 159 150 50 0 0,5 0,5:2,5 160 150 50 10 0,25 0,5:2,5 161 150 50 0 0,25 0,5:2,5 162 150 50 10 0,1 0,5:2,5 163 150 50 0 0,1 0,5:2,5 176 150 50 10 0,5 1:2 177 150 50 0 0,5 1:2 178 150 50 10 0,25 1:2 179 150 50 0 0,25 1:2 180 150 50 10 0,1 1:2 181 150 50 0 0,1 1:2 182 150 50 10 0,5 0,5:2,5 183 150 50 0 0,5 0,5:2,5 184 150 50 10 0,25 0,5:2,5 185 150 50 0 0,25 0,5:2,5 186 150 50 10 0,1 0,5:2,5 187 150 50 0 0,1 0,5:2,5 176 150 50 10 0,5 1:2 177 150 50 0 0,5 1:2 178 150 50 10 0,25 1:2 179 150 50 0 0,25 1:2 180 150 50 10 0,1 1:2 181 150 50 0 0,1 1:2 182 150 50 10 0,5 0,5:2,5 183 150 50 0 0,5 0,5:2,5 184 150 50 10 0,25 0,5:2,5 185 150 50 0 0,25 0,5:2,5 186 150 50 10 0,1 0,5:2,5 187 150 50 0 0,1 0,5:2,5

[0078] The average particle size and size distribution were analyzed and the results are shown in Table 15: Table 15. formulation average particle size (nm) PDI 152 244,8 0,263 153 196,6 0,227 154 249,2 0,263 155 206,9 0,256 156 259,9 0,303 157 199,6 0,203 158 246,3 0,366 159 767 0,313 160 231,8 0,424 161 323,1 0,452 162 228,9 0,392 163 293,8 0,392 176 245,2 0,199 177 236,2 0,261 178 227,5 0,231 179 199 0,219 180 229 0,192 181 192,1 0,157 182 214,8 0,39 183 322,4 0,479 184 221,5 0,363 185 292,7 0,374 186 225,9 0,305 187 292,7 0,318 176 239,1 0,242 177 202,9 0,233 178 233,3 0,237 179 190,7 0,234 180 252,5 0,237 181 196,2 0,219 182 223,6 0,304 183 352,4 0,519 184 226,2 0,292 185 312,7 0,284 186 241 0,372 187 301,6 0,412 Example 11:

[0079] Further nanoparticles containing the active ingredient as in Example 7 were prepared according to the invention. Table 16 shows the test conditions: Table 16. formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Non-Solvent Solvent / Non-Solvent Ratio Active ingredient Vitamin E Deoxycholic acid Lipoid 5100 395 50 2,5 10 50,0 HCl pH 3.5 0,3 399 2,5 2,5 10,0 400 30,0 50,0 5,0

[0080] The average particle size and size distribution were analyzed and the results are shown in Table 17: Table 17. formulation average particle size [nm] PDI 395 397,4 0,188 399 322,9 0,505 400 471,9 1,000 Example 12:

[0081] Further nanoparticles containing the active ingredient as in Example 7 were prepared according to the invention. Table 18 shows the test conditions: Table 18. formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Concentration in non-solvent [mg / mL] Solvent / Non-Solvent Ratio Active ingredient Vitamin E Lipoid 5100 Non-Solvent Sodium deoxycholate 383 50 3,0 150 HCl pH 3.5 25,0 0,5

[0082] The average particle size and size distribution were analyzed and the results are shown in Table 19: Table 19. formulation average particle size [nm] PDI Free active ingredient (%) 383 527,7 0,304 4,5 Example 13:

[0083] Further nanoparticles containing the active ingredient as in Example 7 were prepared according to the invention. Table 20 shows the test conditions: Table 20. formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Concentration in non-solvent [mg / mL] Solvent / Non-Solvent Ratio Active ingredient Vitamin E Lipoid 5100 Non-Solvent Sodium oleate 506 50 2,5 200 Water 5,0 0,5 507 2,5 250 5,0 508 5,0 200 5,0 509 5,0 250 5,0 510 2,5 200 10,0 511 2,5 250 10,0 512 5,0 200 10,0 513 5,0 250 10,0

[0084] The average particle size and size distribution were analyzed and the results are shown in Table 21: Table 21. formulation average particle size [nm] PDI Free active ingredient (%) 506 520,6 0,464 5,7 507 519,0 0,659 4,5 508 574,4 0,381 4,8 509 571,5 0,408 3,0 510 347,0 0,367 0,8 511 373,2 0,451 0,7 512 307,8 0,359 0,7 513 388,5 0,381 0,3 Example 14:

[0085] Further nanoparticles containing the active ingredient as in Example 7 were prepared according to the invention. Table 22 shows the test conditions: Table 22. formulation Concentration in solvent (85:15 EtOH:H2O) [mg / mL] Concentration in non-solvent (water) [mg / mL] Solvent / Non-Solvent Ratio Active ingredient Vitamin E Lipoid 5100 Sodium deoxycholate Sodium oleate 492 50 2,5 50 25,0 5,0 0,5 493 100 494 150 504 200 505 250

[0086] The average particle size and size distribution were analyzed and the results are shown in Table 23: Table 23. formulation average particle size [nm] PDI Free active ingredient (%) 492 834,3 0,675 - 493 691,4 0,069 23,7 494 501,3 0,201 24,6 504 475,1 0,084 38,0 505 504,2 0,627 25,4

Claims

1. A process for the preparation of a carrier system, wherein the carrier system comprises a pharmaceutically active agent and at least one glycerophospholipid, the process comprising the steps of: - providing a solvent stream comprising an organic solvent, the at least one glycerophospholipid and the pharmaceutically active agent, - providing an aqueous stream, and - mixing the solvent stream and the aqueous stream by frontal collision, whereby the carrier system is precipitated as nanoparticles, characterized in that the average particle size of the precipitated nanoparticles measured by dynamic light scattering (DLS) at a temperature of 25 °C is 10 to 500 nm, the solvent stream contains 1 to 1000 mg / mL of the at least one glycerophospholipid and the hydrodynamic pressure in the aqueous stream is at least 1 bar.

2. The process according at least to claim 1, characterized in that the average particle size of the precipitated nanoparticles measured by dynamic light scattering (DLS) at a temperature of 25 °C is 25 to 500 nm, preferably 50 to 250 nm, 70 to 170 nm, 80 to 150 nm, or 90 to 110 nm.

3. The process according to at least claim 1 or 2, characterized in that the hydrodynamic pressure in the aqueous stream is at least 2 bar, preferably 2 bar to 60 bar.

4. The process according to at least one of the preceding claims, characterized in that the hydrodynamic pressure in the solvent stream is at least 0.01 bar, preferably greater than 0.1 bar, more preferably greater than 1 bar, even more preferably between 2 bar and 60 bar.

5. The process according to at least one of the preceding claims, characterized in that the average particle size of the precipitated nanoparticles is adjusted by the hydrodynamic pressure in the aqueous stream and / or the hydrodynamic pressure in the solvent stream, in particular while maintaining a constant mixing ratio of the aqueous phase and solvent phase, whereby nanoparticles of smaller average particle size are precipitated by a higher hydrodynamic pressure.

6. The process according to at least one of the preceding claims, characterized in that the at least one glycerophospholipid is selected from phosphatidylcholine.

7. The process according to at least one of the preceding claims, characterized in that the solvent stream contains 50 to 700 mg / mL, 100 to 500 mg / mL, 100 to 300 mg / mL, or about 150 or 225 mg / mL, of the at least one glycerophospholipid.

8. The process according to at least one of the preceding claims, characterized in that the solvent stream and / or the aqueous stream contains 1 to 250 mg / mL, preferably 5 to 100 mg / mL, more preferably 8 to 50 mg / mL, of a steroid, preferably cholesterol or one of its derivatives.

9. The process according to at least one of the preceding claims, characterized in that the mixing of the solvent stream and the aqueous stream comprises colliding the streams, preferably at a volume ratio of the solvent stream to the aqueous stream of between 1:1 and 1:20, preferably between 1:1 and 1:10.

10. The process according to at least one of the preceding claims, characterized in that the aqueous stream contains 0.1 to 250 mg / mL, preferably 0.5 to 100 mg / mL, more preferably 1.0 to 15 mg / mL, of one or more ionic or nonionic surfactants, the surfactant preferably being sodium oleate and / or oleic acid.

11. The process according to at least one of the preceding claims, characterized in that the solvent stream contains 0.1 to 250 mg / mL, preferably 0.5 to 100 mg / mL, more preferably 1.0 to 15 mg / mL, of one or more ionic or nonionic surfactants, the surfactant preferably being sodium oleate and / or oleic acid.

12. The process according to at least one of the preceding claims, characterized in that the organic solvent comprises a water-miscible solvent, wherein the organic solvent is preferably i) ethanol or ii) a mixture of ethanol with a further organic solvent which is miscible with water and ethanol.

13. The process at least according to claim 12, characterized in that the further organic solvent is acetone.

14. The process according to at least one of the preceding claims, characterized in that the aqueous stream contains 1 to 1000 mM (acid and conjugate base), preferably 20 to 500 mM (acid and conjugate base), of a buffer system.

15. The process according to at least one of the preceding claims, characterized in that the concentration (mg / mL) of the active pharmaceutical ingredient in the solvent stream is lower than the concentration (mg / mL) of the glycerophospholipid, wherein the solvent stream preferably contains 0.25 to 250 mg / mL, preferably 1 to 150 mg / mL, preferably 10 to 100 mg / mL, of the active pharmaceutical ingredient.