ACTIVE SUBSTANCE DELIVERY SYSTEM

Abstract

The present invention relates to an active substance delivery system, preferably an anti-cancer agent delivery system, comprising one or more active substances included in nanoparticles and a method for producing such a delivery system.

Claims

1-13. (canceled)

14. A method for treating cancer in a human subject comprising administering an active substance delivery system to the human subject, wherein the delivery system comprises: (a) nanoparticles comprising: (1) 0.05 to 15 wt.-%, based on the total weight of the nanoparticles, of one or more active substances chosen from cancer chemotherapeutic agents, immunomodulatory drugs, antibodies, viruses, vitamins, enzymes, and combinations thereof, (2) 75 to 99.9 wt.-%, based on the total weight of the nanoparticles, of poly(lactic-co-glycolic acid) (PLGA), and (3) optionally, 0.5 to 20 wt.-%, based on the total weight of the nanoparticles, of one or more surface coating materials chosen from chitosan, carboxymethyl chitosan, cellulose, starch, polyethylene glycols (PEG), polyvinyl alcohols, polymers or block co-polymers, dextran, albumin, surfactants, and pullulan; wherein the average size of the nanoparticles is in a range of from 100 to 350 nm; and (b) optionally, one or more further pharmaceutically acceptable components.

15. The method of claim 14, wherein the weight ratio of (a)(2) to (a)(1) is from 2:1 to 150:1.

16. The method of claim 14 comprising the one or more further pharmaceutically acceptable components of (b), wherein the one or more further pharmaceutically acceptable components are chosen from carriers, polymers, surfactants, stabilizers, wetting agents, emulsifiers, antioxidants, pH influencing agents, disintegrants, recrystallization agents, fluxing agents, preservatives, solvents, salts fillers, binders, foamers, defoamers, lubricants, adsorbents for adjusting the osmotic pressure, and buffers.

17. The method of claim 14, wherein the delivery system is prepared by: (i) dissolving PLGA in a solvent, (ii) dissolving one or more active substances in a solvent, (iii) mixing the solutions of (i) and (ii) together, (iv) providing a polyvinyl alcohol (PVA) solution, (v) adding the mixture of (iii) to the PVA solution by injection to produce nanoparticles, wherein amounts of the mixture and the solution are such that a ratio of a least 1 ml of solution per 5 mg of the summed weight of PLGA and the one or more active substances is achieved, (vi) optionally, evaporating the solvent of (i) and/or (ii), (vii) purifying the nanoparticles and subsequently dispersing the nanoparticles in a dispersant, (viii) coating the nanoparticles with one or more surface coating materials, and (ix) optionally, adding one or more further pharmaceutically acceptable components to the nanoparticles.

18. The method of claim 14, wherein the solvent of (i) and/or the solvent of (ii) comprises acetone, ethyl acetate, a chlorinated solvent, DMSO, a methylated solvent, tetrahydrofuran, a halogenated hydrocarbon, a dioxane, acetonitrile, or a combination thereof and/or the PVA solution of (v) comprises water, ethanol, saline, or a combination thereof and/or the dispersant of (vii) comprises saline, glucose, a surfactant, a pH modifier, water, or a combination thereof.

19. The method of claim 14, wherein the one or more active substances are chosen from: chemotherapeutic agents used against a cancer type chosen from gastrointestinal cancer, gastric cancer, colorectal cancer, hepatobiliary or pancreatic cancer, appendix cancer, esophageal cancer, hepatocellular carcinoma, primary peritoneal cancer, ovarian cancer, endometrial cancer, prostate cancer, leukaemia, lymphoma, soft-tissue sarcoma, multiple myeloma, bladder cancer, lung cancer, thyroid cancer, Kaposi's sarcoma, tumours of embryonal origin; and/or immunomodulatory drugs chosen from curcumin, lutein, hesperidin, apigenin, hesperetin, ajoene, arctigenin, β-carotene, epigallocatechin-3-gallate, glabridin, and quinic acid.

20. The method of claim 14, wherein the method treats a cancer type chosen from gastrointestinal cancer, gynaecological cancer, prostate cancer, peritoneal metastasis, leukaemia, lymphoma, soft-tissue sarcoma, multiple myeloma, breast cancer, bladder cancer, lung cancer, thyroid cancer, Kaposi's sarcoma, ovarian cancer, peritoneal metastasis colorectal cancer, and stomach cancer.

21. The method of claim 14, wherein the method is performed with an assisting tool chosen from microneedles, spray devices, angio-injectors, and a combination thereof.

22. The method of claim 14, wherein a single administration of the delivery system delivers 120 mg to 2500 mg of the one or more active substances to the human subject and/or delivers 30 mg/m.sup.2 body surface to 250 mg/m.sup.2 body surface of the one or more active substances to the human subject.

23. A method for producing an active substance delivery system comprising: (i) dissolving PLGA in a solvent, (ii) dissolving one or more active substances in a solvent, (iii) mixing the solutions of (i) and (ii) together, (iv) providing a polyvinyl alcohol (PVA) solution, (v) adding the mixture of (iii) to the PVA solution by injection to produce nanoparticles, wherein amounts of the mixture and the solution are such that a ratio of a least 1 ml of solution per 5 mg of the summed weight of PLGA and the one or more active substances is achieved, (vi) optionally, evaporating the solvent of (i) and/or (ii), (vii) purifying the nanoparticles and subsequently dispersing the nanoparticles in a dispersant, (viii) coating the nanoparticles with one or more surface coating materials, and (ix) optionally, adding one or more further pharmaceutically acceptable components to the nanoparticles.

24. The method of claim 23, wherein: the solvent of (i) and/or the solvent of (ii) comprises acetone, ethyl acetate, a chlorinated solvent, DMSO, a methylated solvent, tetrahydrofuran, a halogenated hydrocarbon, a dioxane, acetonitrile, or a combination thereof; and/or the PVA solution of (v) comprises water, ethanol, saline, or a combination thereof; and/or the dispersant of (vii) comprises saline, glucose, a surfactant, a pH modifier, water, or a combination thereof.

25. The method of claim 23, wherein the one or more active substances are chosen from: chemotherapeutic agents used against a cancer type chosen from gastrointestinal cancer, particularly gastric cancer, colorectal cancer, hepatobiliary or pancreatic cancer, appendix cancer, esophageal cancer, hepatocellular carcinoma, primary peritoneal cancer, ovarian cancer, endometrial cancer, prostate cancer, leukaemia, lymphoma, soft-tissue sarcoma, multiple myeloma, bladder cancer, lung cancer, thyroid cancer, Kaposi's sarcoma, tumours of embryonal origin; and/or immunomodulatory drugs chosen from curcumin, lutein, hesperidin, apigenin, hesperetin, ajoene, arctigenin, β-carotene, epigallocatechin-3-gallate, glabridin, and quinic acid.

26. The method of claim 23 comprising the addition of the one or more further pharmaceutically acceptable components of (ix) to the nanoparticles, wherein the one or more further pharmaceutically acceptable components are chosen from carriers, polymers, surfactants, stabilizers, wetting agents, emulsifiers, antioxidants, pH influencing agents, disintegrants, recrystallization agents, fluxing agents, preservatives, solvents, salts fillers, binders, foamers, defoamers, lubricants, adsorbents for adjusting the osmotic pressure, and buffers.

Description

[0087] FIG. 1 is a picture of exemplary nanoparticles as described herein, consisting of PLGA and loaded with paclitaxel, obtained with a scanning electron microscope (SEM).

[0088] FIG. 2 is a picture of exemplary nanoparticles as described herein, consisting of PLGA and loaded with curcumin, obtained with a scanning electron microscope (SEM).

[0089] Preferred embodiments and further aspects of the present invention also emerge from the attached patent claims and the following examples, wherein the present invention is not limited to these examples.

EXAMPLES

Example 1: Chemotherapeutic Agent

[0090]

TABLE-US-00001 Amount [mg] Ingredient A B C PLGA 24.75 24.50 24.75 Paclitaxel 0.25 0.50 0.25 Acetone 1.375 1.250 4.875 Acetonitrile 0.125 0.250 0.125

[0091] PLGA was provided and dissolved in acetone. Additionally, paclitaxel was provided separately and dissolved in acetonitrile. Both solutions were mixed.

[0092] 10 ml of a 2% polyvinyl alcohol (PVA) solution were prepared, wherein the polyvinyl alcohol was dissolved in water.

[0093] The mixture obtained above was added to the PVA solution by injection with a syringe pump with a constant rate of 0.5 ml/min. Upon addition, nanoparticles were produced.

[0094] The nanoparticles were further purified by 2×20 min of centrifugation at 10.000×g and at 20° C. The supernatant was removed and the pelleted nanoparticles were redispersed in 5 ml of water and 0.25 g of glucose.

[0095] The nanoparticles had an average size (diameter) of 146 to 217 nm.

Example 2: Immunomodulatory Drug

[0096]

TABLE-US-00002 Amount [mg] Ingredient D E F G H PLGA 39.6 39.6 29.5 95 99 Curcumin 0.4 0.4 0.5 5 1 Acetone — — 2 — 4 Acetonitrile 4 3 — 6 — Chitosan — — — — 6

[0097] PLGA was provided and dissolved in acetone or acetonitrile. Additionally, curcumin was provided separately.

[0098] For examples 2D to 2F, 20 ml of a 2% polyvinyl alcohol (PVA) solution were prepared, wherein the polyvinyl alcohol was dissolved in water. Furthermore, for example 2H, chitosan was additionally added to the PVA solution for coating the nanoparticles.

[0099] For examples 2G and 2H, 40 ml of a 2% polyvinyl alcohol (PVA) solution were prepared, wherein the polyvinyl alcohol was dissolved in water.

[0100] The mixture obtained above was added to the PVA solution by injection with a syringe pump. Examples 2D, 2F, 2G and 2H were added at a constant rate of 0.5 ml/min. Example 2E was added at a constant rate of 1 ml/min. Upon addition, nanoparticles were produced. Furthermore, the nanoparticles were coated with chitosan.

[0101] The nanoparticles were further purified by 3×10 min of centrifugation (Examples 2D to 2F), by 3×15 min of centrifugation (Example 2G) or by 3×20 min of centrifugation (Example 2H) at 10.000×g (Examples 2D and 2E) or at 15.000×g (Examples 2F to 2H) and at 20° C. The supernatant was removed and the pelleted nanoparticles were redispersed in 20 ml of water.

[0102] The nanoparticles of examples 2D, 2F, 2G and 2H had an average size (diameter) of 189 to 231 nm. The included nanoparticles of examples 2E had an average size (diameter) of 221 to 233 nm.

Example 3: Chemotherapeutic Agent and Immunomodulatory Drug

[0103]

TABLE-US-00003 Amount [mg] Ingredient I J K PLGA 49.00 49.25 49.45 Paclitaxel 0.50 0.50 0.50 Curcumin 0.50 0.25 0.05 Acetone 4.975 4.975 4.975 Acetonitrile 0.025 0.025 0.025

[0104] PLGA was provided and dissolved in acetone. Additionally, paclitaxel and curcumin were provided separately and dissolved in acetonitrile.

[0105] 10 ml of a 2% polyvinyl alcohol (PVA) solution were prepared, wherein the polyvinyl alcohol was dissolved in water.

[0106] The mixture obtained above was added to the PVA solution by injection with a syringe pump at a constant rate of 0.5 ml/min to produce nanoparticles.

[0107] The nanoparticles were further purified by 3×15 min of centrifugation at 15.000×g and at 20° C. The supernatant was removed and the pelleted nanoparticles were redispersed in 20 ml of water.

[0108] The nanoparticles had an average size (diameter) of 177 to 207 nm.

Example 4: Spraying of Preheated Unloaded Nanoparticles

[0109] The following experiment was performed to examine an effect of preheating and spraying on nanoparticle properties.

[0110] Therefore, several batches of unloaded nanoparticles were prepared:

[0111] For each batch of unloaded nanoparticles 500 mg PLGA was provided and dissolved in 30 mL acetonitrile.

[0112] 200 mL of a 2% polyvinyl alcohol (PVA) solution were prepared, wherein the polyvinyl alcohol was dissolved in water.

[0113] The polymer solutions obtained above were added to the PVA solution by injection with a syringe pump at a constant rate of 0.5 ml/min to produce nanoparticles.

[0114] Optionally: The solvents are evaporated overnight.

[0115] The nanoparticles were further purified by 3×15 min of centrifugation at 15.000×g and at 20° C. The supernatant was removed, and the pelleted nanoparticles were redispersed in 2 ml of water. All batches were pooled.

[0116] The nanoparticles had a size (diameter) of 185 nm.

[0117] Nanoparticle concentrations of 1, 0.75 and 5% were prepared as 30 ml triplicates and were preheated to 30 degrees while stirring. Then the preheated particles were sprayed with a flow rate of 0.5 ml/s through a nebulizer onto water (MilliQ) in a distance of 5 cm. Spraying was performed with a heated cuff at room temperature (22° C.).

[0118] Zeta sizer measurements of the heated as well as the heated and sprayed samples as described showed no significant change in particle properties.

Example 5: Spraying of Unloaded and Curcumin-Loaded Nanoparticles

[0119] The following experiment was performed to examine an effect of spraying on properties of unloaded and curcumin-loaded nanoparticles.

[0120] Therefore, several batches of unloaded and curcumin loaded nanoparticles were prepared:

[0121] For each batch curcumin loaded nanoparticles 500 mg PLGA was provided and dissolved in 29.5 ml acetonitrile. Additionally, 5 mg per batch curcumin was provided separately and dissolved in 0.5 ml acetonitrile and mixed to the PLGA-acetonitrile mixture. For each batch of unloaded nanoparticles 500 mg PLGA were provided and dissolved in 30 ml acetonitrile.

[0122] 200 ml of a 2% polyvinyl alcohol (PVA) solution were prepared, wherein the polyvinyl alcohol was dissolved in water.

[0123] The polymer solutions obtained above was added to the PVA solution by injection with a syringe pump at a constant rate of 0.5 ml/min to produce nanoparticles.

[0124] Optionally: The solvents are evaporated overnight.

[0125] The nanoparticles were further purified by 3×15 min of centrifugation at 15.000×g and at 20° C. The supernatant was removed, and the pelleted nanoparticles were redispersed in 2 ml of water. All batches were pooled.

[0126] The nanoparticles had a size (diameter) of 177 nm (unloaded) and 184 nm (curcumin loaded).

[0127] Nanoparticle concentrations of 1, 0.75 and 5% were prepared as 30 ml triplicates and were sprayed with a flow rate of 0.5 ml/s through a nebulizer onto water (MilliQ) in a distance of 5 cm. Spraying was performed at room temperature (22° C.).

[0128] Zeta sizer measurements of the sprayed samples as described showed no significant change in particle properties by spraying.