METHOD FOR PREPARING BIODEGRADABLE POLYMER MICROPARTICLES, AND BIODEGRADABLE POLYMER MICROPARTICLES PREPARED THEREBY

Abstract

Provided are: a method for preparing biodegradable polymer microparticles and, particularly, porous microparticles of a biodegradable polymer; and biodegradable polymer microparticles prepared thereby, particularly, porous microparticles of a biodegradable polymer.

Claims

1. A method for preparing biodegradable polymer microparticles, the method comprising the steps of: 1) dissolving a biodegradable polymer in an organic solvent to form a dispersed phase; 2) applying pressure to the dispersed phase formed in step 1) to allow the dispersed phase to pass through the pore of SPG membrane (Shirasu Porous Glass membrane), and thereby to form an emulsion in a continuous phase comprising a surfactant; and 3) removing the organic solvent from the emulsion formed in step 2) to form microparticles.

2. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the microparticles satisfy the following features i) and ii): i) spherical shape; ii) particle diameter of 10 to 200 m.

3. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the dispersed phase further comprises porogen to form porous microparticles.

4. The method for preparing biodegradable polymer microparticles according to claim 3, wherein the porous microparticles satisfy the following features i) to iv): i) spherical shape; ii) particle diameter of 10 to 200 m; iii) pore diameter of 0.1 to 20 m; iv) porosity ratio of 10 to 50%.

5. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the microparticles have d.sub.10 of greater than 20 m and d.sub.90 of less than 100 m.

6. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the microparticles have a span value of less than 1.

7. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the SPG membrane has pores ranging from 10 to 30 m.

8. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the pressure applied to the dispersed phase is in a range of from 0.1 to 10 kPa.

9. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the surfactant has a concentration in a range of from 0.5 to 5% by weight.

10. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the concentration of biodegradable polymer dissolved in the dispersed phase is in a range of from 5 to 20% by weight.

11. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the continuous phase is agitated at a speed in a range of from 100 to 500 rpm.

12. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the biodegradable polymer is one or more selected from the group consisting of poly(lactic acid), poly(glycolic acid), poly(dioxanone), poly(caprolactone), poly(lactic acid-co-glycolic acid), poly(dioxanone-co-caprolactone), poly(lactic acid-co-caprolactone), derivatives thereof and copolymers thereof.

13. The method for preparing biodegradable polymer microparticles according to claim 1, wherein the biodegradable polymer has a number average molecular weight (Mn) in a range of 10,000 to 1,000,000 g/mol.

14. Biodegradable polymer microparticle prepared by the method according to claim 1.

15. A method for preparing polymer filler, the method comprising the steps of: preparing biodegradable polymer microparticles by the method according to claim 1; and mixing the biodegradable polymer microparticles with one or more biocompatible carriers.

16. Polymer filler comprising: biodegradable polymer microparticle according to claim 14; and one or more biocompatible carriers.

Description

BRIEF EXPLANATION OF THE DRAWINGS

[0024] FIG. 1 shows scanning electron microscope (SEM) photographs of the biodegradable polymer porous microparticles prepared according to Example 1 of the present invention.

[0025] FIG. 2 shows scanning electron microscope (SEM) photographs of the biodegradable polymer porous microparticles prepared according to Comparative Example 1 of the present invention.

[0026] FIG. 3 shows the sizes and particle size distribution of the biodegradable polymer porous microparticles prepared according to Example 1 of the present invention.

[0027] FIG. 4 shows the sizes and particle size distribution of the biodegradable polymer porous microparticles prepared according to Comparative Example 1 of the present invention.

[0028] FIG. 5 is a schematic drawing of the device for preparing porous microparticles by using SPG membrane according to Example 1 of the present invention.

[0029] FIG. 6 shows scanning electron microscope (SEM) photographs of the SPG membrane used in Example 1 of the present invention.

[0030] FIG. 7 shows DSLR (Digital Single Lens Reflex) camera photographs of the injection parts of mice after injecting the polymer filler prepared in Example 4 and the filler of Comparative Example 2 of the present invention.

[0031] FIG. 8 shows three-dimensional photographs of the injection parts of mice after injecting the polymer filler prepared in Example 4 and the filler of Comparative Example 2 of the present invention.

DETAILED DESCRIPTION

[0032] The present invention is explained in detail below.

[0033] In an embodiment of the present invention, the biodegradable polymer microparticles can satisfy the following features i) and ii):

[0034] i) spherical shape,

[0035] ii) particle diameter of 10 to 200 m,

[0036] Preferably, the particle size of the polymer microparticle is smaller than the diameter of the injection needle to be used so that it can be injected, and preferably the shape of the particle is substantially in spherical form so as not to cause pain to the patient and not to be felt by touch. In an embodiment, the particle size (particle diameter) of the biodegradable polymer microparticle can be typically 200 m or less, and it preferably has a diameter of 10 m or greater in order not to be taken by macrophage in living tissues. In a preferable embodiment, the microparticle can have a diameter of 10 to less than 100 m, more preferably 10 to 80 m, still more preferably 10 to 50 m, and most preferably 20 to 40 m.

[0037] In an embodiment of the present invention, the dispersed phase can further comprise porogen so as to form porous microparticles. In a preferable embodiment, the concentration of porogen dissolved in an organic solvent can be in a range of from 1 to 10% by weight. If the concentration of porogen is lower than 1% by weight, pores may not be formed sufficiently. If the concentration of porogen is higher than 10% by weight, due to the excessive pore formation, the physical strength of the porous microparticle itself becomes weak and cells may permeate into the pores.

[0038] In a preferable embodiment, the porous microparticle has the features described in Korean Patent Application No. 2016-0169309. Korean Patent Application No. 2016-0169309 is herein incorporated by reference in its entirety. For instance, the porous microparticles can satisfy the following features i) to iv):

[0039] i) spherical shape,

[0040] ii) particle diameter of 10 to 200 m,

[0041] iii) pore diameter of 0.1 to 20 m, and

[0042] iv) porosity ratio of 10 to 50%.

[0043] In an embodiment, as the standard of particle size distribution, the porous microparticle of biodegradable polymer has d.sub.10 of greater than 20 m and d.sub.90 of less than 100 m, preferably d.sub.10 of greater than 20 m and d.sub.90 of less than 60 m, and still more preferably d.sub.10 of greater than 25 m and d.sub.90 of less than 40 m.

[0044] Also, in an embodiment, the porous microparticle of biodegradable polymer should have a span value, which shows uniform distribution of particles, of less than 1, preferably less than 0.8, and more preferably less than 0.6. The span value becomes greater as the particle size distribution becomes broad, and it becomes close to 0 as the particle size distribution becomes narrow. The span value is calculated by the following equation:

[00001]$\mathrm{span}=\frac{D_{90}-D_{10}}{D_{50}}$

[0045] [Definitions of D.sub.10, D.sub.50 and D.sub.90: Size values corresponding to 10%, 50% and 90%, respectively, of the maximum value in accumulated distribution of particles, represented as the particle sizes corresponding to 1/10, 5/10 and 9/10, respectively, of the particle size distribution curve showing the relatively accumulated amounts of particles according to the size, when it is measured, plotted and divided into 10 fractions.]

[0046] The porous microparticle according to an embodiment of the present invention can have a larger volume per the same mass according to the porosity ratio. In an embodiment, the porosity ratio of the porous microparticle of biodegradable polymer can be 5 to 50%, preferably 10 to 50%, and more preferably 10 to 30%.

[0047] In the present invention, the porosity ratio is obtained according to the following equation:

Porosity ratio=(Volume of porous polymer microparticleVolume of non-porous polymer microparticle)/Volume of porous polymer microparticle100

[0048] The pore size (diameter) of the porous microparticle of biodegradable polymer according to the present invention can be 0.1 m to 20 m, and preferably 0.1 to 10 m, and more preferably 1 to 10 m.

[0049] An embodiment of the present invention is characterized in preparing microparticles, particularly porous microparticles, of biodegradable polymer by applying SPG membrane emulsification method. A device for preparing porous microparticles by using SPG membrane according to Example 1 of the present invention is schematically shown in FIG. 5, but the scope of the present invention is not limited thereby.

[0050] In an embodiment of the present invention, the concentration of biodegradable polymer dissolved in an organic solvent is preferably in a range of from 5 to 20% by weight, particularly from 10 to 20% by weight, more particularly from 10 to 15% by weight. If the concentration of biodegradable polymer is lower than 5% by weight, the concentration of dispersed phase becomes low and the viscosity decreases, and it may be difficult to generate the emulsion with uniform size when passing through SPG membrane. If the concentration of biodegradable polymer is higher than 20% by weight, the concentration of dispersed phase becomes high and the viscosity increases, and not uniformly-sized emulsion but thread-like polymer may be formed when passing through SPG membrane.

[0051] In an embodiment of the present invention, SPG membrane can have pores ranging from 10 to 30 m, particularly from 10 to 20 m, and more particularly from 15 to 20 m. If the pore size of SPG membrane is smaller than 10 m, the average size of the formed porous microparticles is smaller than 20 m and thus they can be taken by macrophage. If the pore size of SPG membrane is larger than 30 m, the average size of the formed porous microparticles is larger than 50 m and thus clogging of injection needle may be caused. FIG. 6 shows scanning electron microscope (SEM) photographs of the SPG membrane used in the example of the present invention, but the scope of the present invention is not limited thereby.

[0052] In an embodiment of the present invention, the pressure applied to the dispersed phase can be in a range of from 0.1 to 10 kPa, particularly from 1 to 10 kPa, and more particularly from 2 to 7 kPa. If the pressure is lower than 0.1 kPa, the dispersed phase cannot pass through SPG membrane, and thus the emulsion may not be formed. If the pressure is higher than 10 kPa, the dispersed phase passes through SPG membrane too fast, and thus not uniformly-sized emulsion but thread-like polymer may be formed.

[0053] In an embodiment of the present invention, the surfactant can have a concentration in a range of from 0.5 to 5% by weight, particularly from 0.5 to 3% by weight, and more particularly from 1 to 3% by weight. If the surfactant concentration is lower than 0.5% by weight, porous microparticles may be generated with non-uniform particle size. If the surfactant concentration is higher than 5% by weight, porous microparticles may be made with non-uniform pore size. In an embodiment, the surfactant can be selected from polyvinyl alcohol, sodium dodecyl sulfate, polysorbate, polyethylene glycol and combinations thereof. Preferably, polyvinyl alcohol can be used.

[0054] In an embodiment of the present invention, the continuous phase can be agitated at a speed in a range of from 100 to 500 rpm, particularly from 100 to 300 rpm, and more particularly from 100 to 250 rpm. If the agitation speed is lower than 100 rpm, the emulsion is dewy on SPG membrane and the dews get bigger thereon without falling, and thus porous microparticles may be formed with an average particle size of greater than 50 m. If the agitation speed is higher than 500 rpm, the emulsion dews on SPG membrane fall too fast, and thus porous microparticles may be formed with an average particle size of less than 20 m.

[0055] In a preferable embodiment, it is possible to use the combination of two or more of, particularly all of, the above specific ranges of biodegradable polymer concentration, pore size of SPG membrane, pressure applied to the dispersed phase and surfactant concentration in continuous phase, and in addition thereto the agitation speed of continuous phase.

[0056] In an embodiment of the present invention, the biodegradable polymer can be one or more selected from the group consisting of poly(lactic acid), poly(glycolic acid), poly(dioxanone), poly(caprolactone), poly(lactic acid-co-glycolic acid), poly(dioxanone-co-caprolactone), poly(lactic acid-co-caprolactone), derivatives thereof and copolymers thereof. Preferably, the biodegradable polymer is poly(lactic acid) or poly(caprolactone), and more preferably poly(caprolactone).

[0057] In an embodiment of the present invention, the biodegradable polymer can have a number average molecular weight (Mn) in a range of from 10,000 to 1,000,000 g/mol. More preferably, it can be in a range of from 10,000 to 100,000 g/mol.

[0058] According to an embodiment of the present invention, the amount of the porous microparticle of biodegradable polymer contained in the polymer filler can be typically 10 to 50% by weight, and more concretely 10 to 30% by weight, based on 100% by weight of the polymer filler, and it can be adjusted according to the desired volume effect of the desired injection part.

[0059] The polymer filler according to an embodiment of the present invention can further comprise one or more biocompatible carriers. Such a carrier is absorbed in body typically within 1 day to 6 months after the injection.

[0060] In an embodiment, a carrier selected from carboxymethyl cellulose, hyaluronic acid, dextran, collagen and combinations thereof can be used as the biocompatible carrier.

[0061] The amount of the biocompatible carrier contained in the polymer filler of the present invention can be typically 50 to 90% by weight, and more concretely 70 to 90% by weight, based on 100% by weight of the polymer filler.

[0062] As well as the ingredients explained above, additive ingredientsfor example, a lubricant such as glycerin, phosphate buffer or the like conventionally comprised in an injection formulationcan be further comprised in the biocompatible carrier.

[0063] The polymer filler according to an embodiment of the present invention can be an injection formulation preferably. An injection formulation of the polymer filler according to an embodiment of the present invention can be provided as being contained in a sterilized injection syringe or a sterilized vial, and it has high use convenience since no pretreatment is needed, it is safe since 100% thereof is biodegraded over a predetermined time after the injection leaving no foreign substance in living tissues, and it does not cause allergic reaction since it contains no substances derived from animal at all.

[0064] In addition, as compared with the existing polymer product (for example, polymer content of 30%), the polymer filler according to an embodiment of the present invention can provide a greater volume effect with the same amount of polymer, and thus the volume effect can be maintained even if the carrier is absorbed. Therefore, the polymer filler according to an embodiment of the present invention can be used preferably for wrinkle improvement, facial plastic procedure or body plastic procedure.

[0065] The present invention is explained in more detail by the following examples. However, the following examples are intended only to illustrate the present invention, and should not be interpreted as limiting the scope of the present invention in any manner.

EXAMPLE

Example 1

[0066] As a dispersed phase, 5 g of polycarprolactone (PCL) with a number average molecular weight of 45,000 g/mol and 1 g of tetradecane were dissolved in 100 g of methylene chloride. SPG membrane with 15 m pore (FIG. 6) was combined to a SPG membrane emulsification device. The prepared dispersed phase was injected into a continuous phasewhich was 2 wt % aqueous solution of polyvinyl alcoholthrough SPG membrane under nitrogen pressure of 2 kPa. At that time, the aqueous solution of polyvinyl alcohol was agitated at 250 rpm speed, and after completing the injection, the mixture was continuously agitated for 24 hours (FIG. 5). The prepared porous PCL microparticles were agitated in 500 ml of distilled water for 2 hours to wash out the remaining polyvinyl alcohol, recovered by using a centrifuge, and washed with 500 ml of ethyl alcohol for 2 hours to to remove the remaining tetradecane and methylene chloride. After the washing, the porous PCL microparticles were dried in a vacuum dryer for 48 hours to remove the remaining ethyl alcohol.

Example 2

[0067] As a dispersed phase, 1 g of polycarprolactone (PCL) with a number average molecular weight of 50,000 g/mol and 0.2 g of tetradecane were dissolved in 20 g of methylene chloride. SPG membrane with 15 m pore (FIG. 6) was combined to a SPG membrane emulsification device. The prepared dispersed phase was injected into a continuous phasewhich was 2 wt % aqueous solution of polyvinyl alcoholthrough SPG membrane under nitrogen pressure of 2 kPa. At that time, the aqueous solution of polyvinyl alcohol was agitated at 250 rpm speed, and after completing the injection, the mixture was continuously agitated for 24 hours (FIG. 5). The prepared porous PCL microparticles (porosity ratio: 10%) were agitated in 500 ml of distilled water for 2 hours to wash out the remaining polyvinyl alcohol, recovered by using a centrifuge, and washed with 500 ml of ethyl alcohol for 2 hours to to remove the remaining tetradecane and methylene chloride. After the washing, the porous PCL microparticles were dried in a vacuum dryer for 48 hours to remove the remaining ethyl alcohol.

Example 3

[0068] As a dispersed phase, 1 g of polycarprolactone (PCL) with a number average molecular weight of 50,000 g/mol and 0.3 g of tetradecane were dissolved in 20 g of methylene chloride. SPG membrane with 15 m pore (FIG. 6) was combined to a SPG membrane emulsification device. The prepared dispersed phase was injected into a continuous phasewhich was 2 wt % aqueous solution of polyvinyl alcoholthrough SPG membrane under nitrogen pressure of 2 kPa. At that time, the aqueous solution of polyvinyl alcohol was agitated at 250 rpm speed, and after completing the injection, the mixture was continuously agitated for 24 hours (FIG. 5). The prepared porous PCL microparticles (porosity ratio: 20%) were agitated in 500 ml of distilled water for 2 hours to wash out the remaining polyvinyl alcohol, recovered by using a centrifuge, and washed with 500 ml of ethyl alcohol for 2 hours to to remove the remaining tetradecane and methylene chloride. After the washing, the porous PCL microparticles were dried in a vacuum dryer for 48 hours to remove the remaining ethyl alcohol.

Comparative Example 1

[0069] Excepting that the dispersed phase was rapidly fed into 2 wt % aqueous solution of polyvinyl alcohol by using a homogenizer with agitation at 4000 rpm speed and after the agitation for 1 minute the mixture was agitated at 250 rpm speed for 24 hours, porous PCL microparticles were prepared according to substantially the same procedure as Example 1.

[0070] Experimental Example 1 The porous PCL microparticles obtained in Example 1 and the porous PCL microparticles obtained in Comparative Example 1 were observed with scanning electron microscope (SEM). The results are shown in FIG. 1 and FIG. 2, respectively. As can be confirmed from the figures, the present invention using SPG membrane emulsification method could provide porous microparticles with smaller size and narrower particle size distributioni.e., uniform particle sizeas compared with the case using a homogenizer.

Experimental Example 2

[0071] The particle size distributions of the porous PCL microparticles obtained in Example 1 and the porous PCL microparticles obtained in Comparative Example 1 were measured by using a particle size analyzer. The results are shown in FIG. 3 and FIG. 4, respectively. As can be known from the figures, the porous PCL microparticles obtained in Example 1 showed remarkably narrower particle size distributioni.e., uniform particle sizeas compared with the porous PCL microparticles obtained in Comparative Example 1. The results of particle size distribution analysis are shown in the following Table 1.

TABLE-US-00001 TABLE 1 D.sub.10 D.sub.50 D.sub.90 C.V..sup.1) span Example 1 25.35 m 30.17 m 37.57 m 22.2% 0.41 Comparative 14.17 m 31.37 m 63.81 m 50.9% 1.58 Example 1 .sup.1)C.V. (coefficient of variation): The value of dividing standard deviation by average, and the standard for measuring the degree of relative dispersion. As the calculated value is closer to 0, it means that the particles are populated on the average and the degree of dispersion is small.

Examples 4 to 6

[0072] The polymer filler formulations of Examples 4 to 6 were prepared by mixing the porous microparticles of biodegradable polymer prepared in Examples 1 to 3, respectively, with a carrier prepared from 3% by weight of carboxymethyl cellulose, 27% by weight of glycerin and 70% by weight of phosphate buffer. At that time, the mixing ratio was, based on 100% by weight of the mixture, 30% by weight of the porous microparticles and 70% by weight of the carrier.

Comparative Examples 2 and 3

[0073] Commercially available facial filler (Ellanse) using PCL as the raw material was purchased and used as Comparative Example 2. Commercially available facial filler (Sculptra) using polylactic acid (PLA) as the raw material was purchased and used as Comparative Example 3.

Experimental Example 3

[0074] Each of the formulations of Examples 4 to 6 and Comparative Examples 2 and 3 was filled within a syringe and 200 l thereof was injected into the back of a hairless mouse. The sizes of the injection parts were measured, and the size changes were checked continuously with period of a certain time. The results are shown in the following Table 2. In addition, the polymer filler of Example 4 and the polymer filler of Comparative Example 2 were injected into the mice. From the injection parts for 2 weeks, DSLR (Digital Single Lens Reflex) camera photographs were taken and are shown in FIG. 7, and three-dimensional photographs were taken and are shown in FIG. 8.

[0075] As shown in the following Table 2, as for the filler formulation comprising the porous microparticles of biodegradable polymer according to the present invention, it can be confirmed that the initial volume reduction after the procedure was improved remarkably.

TABLE-US-00002 TABLE 2 Comp. Comp. Exam- Exam- Exam- Exam- Exam- ple 4 ple 5 ple 6 ple 1 ple 2 Volume immediately 100% 100% 100% 100% 100% after the procedure Volume after 3 months 100% 100% 100% 80% 60% Volume after 6 months 80% 80% 85% 40% 20%