Patent Application
20220387321
The present invention relates to a drug-containing microsphere used for sustained release injection and a method for preparing the same.
Sustained release injections are generally manufactured by preparing microspheres in which a drug is included and putting them in an injection. Here, the microspheres are prepared to have a drug included therein. When the microsphere-containing injection is injected into the body, the drug is slowly released from the microspheres injected into the body so as to exhibit a pharmaceutical effect continuously.
Therefore, when an injection solution comprising microspheres for sustained release is injected into the body, the pharmacological effect of the drug may be exhibited for a prolonged period of time by allowing the drug to be slowly released from the microspheres in the body.
By such sustained release injection, the number of invasions into the body may be reduced, and patient medication compliance may be improved. For example, when a drug which is to be administered by oral route once or twice a day or by injection once a day is prepared into a sustained release injection containing microspheres as above, the drug may be formulated so that the medicinal effect lasts for 30 days after administration of a single injection.
Therefore, patient medication compliance may be improved by overcoming the inconvenience of having to orally take the drug every day or receive injections every day.
In particular, in the case of patients with dementia or Alzheimer's disease, Parkinson's disease, or a depressive disorder who need to take therapeutic agents or antipsychotics acting on the central nervous system but have difficulty in taking them regularly, it may be possible to provide a very large advantage in terms of patient medication compliance when a drug is formulated into a sustained release dosage form such as a microsphere-containing injection, and administered, for example, once a month.
Currently, microspheres are generally prepared by a solvent evaporation method. The solvent evaporation method is a method of dissolving a polymer material and a drug in a volatile organic solvent, and then evaporating the organic solvent so that the drug is included in the microsphere.
However, microspheres prepared in this way have a problem that the drug is rapidly released in the initial stage when injected into the body because the drug is present on the surface of the microsphere. As such, when the drug is rapidly released in the initial stage of injection, the blood concentration of the drug rises rapidly, which may cause adverse effects to the patient. In particular, when the blood concentration of a drug acting on the central nervous system rises rapidly, adverse effects such as tremor or miosis may suddenly occur.
Therefore, it is important to suppress such initial drug release from the microsphere.
In general, in order to suppress the initial drug release from the microsphere, the surface is to be washed. For example, the drug deposited on the surface of the microsphere prepared by solvent evaporation, etc. may be removed by washing the drug deposited on the surface of the microsphere with an aqueous ethanol solution, or washing with a Tween-based or Span-based surfactant.
However, when washing the surface of microsphere as above, the washing process is complicated, the process time is long, and the process of treating with ethanol or a surfactant may adversely affect the microsphere or the drug itself, and thus such process may not be preferable.
Therefore, it is necessary to develop a novel method for preparing microspheres which may suppress the initial drug release from the microsphere in a short process time with a simple process without significantly affecting the microsphere or the drug included therein.
Accordingly, the present inventors studied a method for suppressing the initial drug release from the microsphere, and as a result, have found that, when the microsphere was prepared by adding a salt to a continuous phase, it is possible to prevent the formation of drug on the surface of the microsphere and also prevent the formation of pores on the surface of the microsphere, thereby preventing the drug from being excessively released in the initial stage when injected into the body.
Therefore, it is an object of the present invention to provide a method for preparing microspheres whose initial drug release is suppressed by simplifying the process and reducing the process time without significantly affecting the microsphere or the drug included therein.
The present invention relates to a method for preparing a sustained release microsphere containing a drug.
Specifically, the method for preparing a microsphere according to the present invention comprises dissolving an active ingredient and a biodegradable polymer in an organic solvent to prepare a dispersed phase; dissolving a salt in water to prepare a continuous phase; mixing and stirring the dispersed phase and the continuous phase to form an emulsion; removing the organic solvent; and drying.
According to the present invention, drug crystals are not produced around or on the surface of the microsphere, and thus it is not necessary to wash the microsphere with ethanol or a surfactant.
In the present invention, as the salt included in the continuous phase, the salt such as sodium chloride (NaCl), potassium chloride (KCl), calcium chloride (CaCl2)), magnesium sulfate (MgSO4), sodium sulfate (Na2SO4), mannitol, ammonium, potassium sulfate, disodium phosphate, dipotassium phosphate, trisodium phosphate, disodium citrate, trisodium citrate, or sodium succinate may be used, and the salt has a concentration of 1 to 10% (w/v), preferably 2 to 8% (w/v) in the continuous phase.
The microsphere according to the present invention is injected into the body having the drug included therein, and thus a biodegradable polymer such as polylactide, poly(lactide-co-glycolide), polyglycolactide and poly(lactide-co-glycolide)glucose may be used.
In the present invention, the continuous phase may be prepared and used to further comprise a water-soluble polymer. For example, at least one selected from the group consisting of polyvinyl alcohol, polysorbate, poloxamer, polyvinylpyrrolidone, polyvinylmethyl ether, and polyvinyl ether may be used. The dispersibility of the emulsion may be maintained by the water-soluble polymer.
As an active ingredient which may be used in the present invention, a substance acting on the central nervous system is more effective in terms of patient medication compliance. As the active ingredient, a substance showing therapeutic activity for dementia such as donepezil, a therapeutic agent for Parkinson's disease such as pramipexole, rotigotine, and ropinirole, an antipsychotic such as risperidone, blonanserin, and rulasidone, or an alcoholism therapeutic drug such as naltrexone, etc. may be used.
In the microsphere of the present invention, the drug may be present in an amount of about 30-50%, preferably 40-50%, with respect to the total weight of the microsphere.
As another aspect of the present invention, the present invention relates to a microsphere prepared by the above preparation method.
As another aspect of the present invention, the present invention relates to a sustained release injection comprising the microsphere as described above.
The injection containing sustained release microspheres according to the present invention prevents the formation of drug crystals around or on the surface of the microsphere and improves the drug entrapment efficiency into the microsphere, thereby preventing the drug from being excessively released in the initial stage when administered. In addition, since the drug is continuously released, it is possible to increase patient medication compliance and reduce adverse effects caused by dose dumping.
Hereinafter, examples will be described in detail to help the understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to explain the present invention more completely to those of ordinary skill in the art.
2 g (amount of drug loaded in microspheres (hereinafter, “drug loading”) is 40%) of donepezil (manufacturer: Neuland Laboratories, India) and 3 g of poly D,L-lactide (Resomer R 203 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 9 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. In addition, 6.25 g of polyvinyl alcohol and 12.5 g of NaCl were dissolved in 1.25 L of water to prepare a continuous phase.
After putting the continuous phase in a double jacket beaker and maintaining the temperature at 10° C. or below using a constant temperature circulating water bath, the dispersed phase (i.e., donepezil-containing polymer solution) was added to the continuous phase (i.e., polyvinyl alcohol and NaCl aqueous solution) and stirred at high speed to form an emulsion.
Then, in order to remove the organic solvent and obtain solidified microspheres, the organic solvent was volatilized at a temperature of 47° C. for 2 hours, and then slowly cooled to 10° C. for 1 hour. After washing the hardened microspheres with water for injection several times, and going through wet filtration using a sieve and freeze-drying, donepezil-containing microspheres were finally obtained.
2.25 g (drug loading: 45%) of donepezil (manufacturer: Neuland Laboratories, India) and 2.75 g of poly D,L-lactide (Resomer R 203 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 8.25 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. In addition, 6.25 g of polyvinyl alcohol and 12.5 g of NaCl were dissolved in 1.25 L of water to prepare a NaCl 1% (w/v) continuous phase. The rest of the preparation method was carried out in the same manner as in example 1 to prepare microspheres.
Except that NaCl was added to be 5% (w/v) while preparing the continuous phase, the rest of the preparation method was carried out in the same manner as in example 1 to prepare microspheres.
Except that NaCl was added to be 10% (w/v) while preparing the continuous phase, the rest of the preparation method was carried out in the same manner as in example 1 to prepare microspheres.
Except that KCl was added instead of NaCl to be 1% (w/v) while preparing the continuous phase, the rest of the preparation method was carried out in the same manner as in example 1 to prepare microspheres.
Except that KCl was added instead of NaCl to be 5% (w/v) while preparing the continuous phase, the rest of the preparation method was carried out in the same manner as in example 1 to prepare microspheres.
2 g (drug loading: 40%) of donepezil (manufacturer: Neuland Laboratories, India) and 3 g of poly D,L-lactide-co-glycolide (PLGA; Resomer RG 753 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 9 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. The rest of the preparation method was carried out in the same manner as in example 1, except that NaCl was added to be 5% by weight while preparing the continuous phase, to prepare microspheres.
2 g (drug loading: 40%) of donepezil (manufacturer: Neuland Laboratories, India) and 3 g of poly D,L-lactide (Resomer R 203 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 9 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. In addition, 6.25 g of polyvinyl alcohol was dissolved in 1.25 L of water to prepare a continuous phase (salts such as NaCl or KCl were not added).
After putting the continuous phase in a double jacket beaker and maintaining the temperature at 10° C. or below using a constant temperature circulating water bath, the dispersed phase (i.e., donepezil-containing polymer solution) was added to the continuous phase (i.e., polyvinyl alcohol aqueous solution) and stirred at high speed to form an emulsion.
Then, in order to remove the organic solvent and obtain solidified microspheres, the organic solvent was volatilized at a temperature of 47° C. for 2 hours, and then slowly cooled to 10° C. for 1 hour. After washing the hardened microspheres with water for injection several times, and going through wet filtration using a sieve and freeze-drying, donepezil-containing microspheres were finally obtained.
2.25 g (drug loading: 45%) of donepezil (manufacturer: Neuland Laboratories, India) and 2.75 g of poly D,L-lactide (Resomer R 203 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 8.25 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. The rest of the preparation method was carried out in the same manner as in comparative example 1 to prepare microspheres.
2 g of donepezil (manufacturer: Neuland Laboratories, India) and 3 g of poly D,L-lactide (Resomer R 203 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 9 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. In addition, 6.25 g of polyvinyl alcohol was dissolved in 1.25 L of water to prepare a continuous phase (salts such as NaCl or KCl were not added).
After putting the continuous phase in a double jacket beaker and maintaining the temperature at 10° C. or below using a constant temperature circulating water bath, the dispersed phase (i.e., donepezil-containing polymer solution) was added to the continuous phase (i.e., polyvinyl alcohol aqueous solution) and stirred at high speed to form an emulsion.
Then, in order to remove the organic solvent and obtain solidified microspheres, the organic solvent was volatilized at a temperature of 47° C. for 2 hours, and then slowly cooled to 10° C. for 1 hour. After washing the hardened microspheres with water for injection several times, and going through first wet filtration using a sieve, microspheres were primarily obtained.
After washing the primarily obtained microspheres with 20% EtOH aqueous solution at 10° C. for 1 hour, and going through wet filtration again using a sieve and freeze-drying, donepezil-containing microspheres were finally obtained.
The preparation method was carried out in the same manner as in comparative example 2 until primarily obtaining donepezil microspheres.
Then, after washing the primarily obtained microspheres with 3% Tween 20 aqueous solution at 10° C. for 1 hour, and going through wet filtration again using a sieve and freeze-drying, donepezil-containing microspheres were finally obtained.
2 g (theoretical drug loading: 40%) of donepezil (manufacturer: Neuland Laboratories, India) and 3 g of poly(D,L-lactide-co-glycolide) (Resomer RG 753 H; manufacturer: Evonik, Germany), which is a biodegradable polymer, were added to 9 g of dichloromethane (manufacturer: Daejung Chemicals & Metals Co., Ltd., South Korea) and completely dissolved by stirring to prepare a polymer solution, which is a dispersed phase. In addition, 6.25 g of polyvinyl alcohol was dissolved in 1.25 L of water to prepare a continuous phase (salts such as NaCl or KCl were not added).
After putting the continuous phase in a double jacket beaker and maintaining the temperature at 10° C. or below using a constant temperature circulating water bath, the dispersed phase (i.e., donepezil-containing polymer solution) was added to the polyvinyl alcohol aqueous solution (continuous phase) and stirred at high speed to form an emulsion.
Then, in order to remove the organic solvent and obtain solidified microspheres, the organic solvent was volatilized at a temperature of 47° C. for 2 hours, and then slowly cooled to 10° C. for 1 hour. After washing the hardened microspheres with water for injection several times, and going through wet filtration using a sieve and freeze-drying, donepezil-containing microspheres were finally obtained.
About 20 mg of microspheres obtained in each of the examples and comparative examples was fixed to an aluminum stub using carbon tape, and coated with platinum under a vacuum level of 0.1 torr and high voltage (10 kV) for 3 minutes. Then, the microspheres were mounted on the main body (SEM stage) of SEM (equipment name: SEC-SNE 4500M Plus A, South Korea) to observe the surface morphology of the microspheres using an image analysis program (mini-SEM).
As can be seen in
However, as can be seen in
As can be seen in
Although the presence of drug crystals was roughly confirmed from the SEM photograph images, in order to quantitatively confirm the amount of drug crystals actually generated outside the microspheres without being included therein while preparing microspheres, the microspheres prepared in each of the examples and comparative examples 1, 1-1 and 3 were washed with 20% EtOH aqueous solution at 10° C. for 1 hour and freeze-dried to remove drug crystals from the surface and the periphery of the microspheres.
Experimental Example 3: Measurement of actual content of donepezil in microspheres, entrapment efficiency, and amount of drug crystals present outside the microspheres
100 mg of microspheres prepared in each of examples 1 to 4 and comparative examples 1 to 3 was completely dissolved in acetonitrile and then diluted with a mobile phase. 20 uL of the diluted solution was injected into HPLC and measured at a detection wavelength of 318 nm.
In addition, 100 mg of microspheres prepared in each of examples 1 to 4 and comparative examples 1, 1-1 and 3 and washed with an aqueous ethanol solution in experimental example 2 was completely dissolved in acetonitrile and then diluted with a mobile phase. 20 uL of the diluted solution was injected into HPLC and measured at a detection wavelength of 318 nm.
Column: Luna phenyl-Hexyl, C18 5 μm, 4.6×250 mm
Mobile phase: pH 2.0 tetrahydrofuran, 3:1 mixed solution of triethylamine solution (solution A) and methanol tetrahydrofuran solution (solution B)
Table 1 shows the measured drug entrapment efficiency (%) and the drug entrapment efficiency in microspheres after washing with ethanol.
Theoretical drug loading amount (%)=Amount of drug introduced while preparing microspheres/(Amount of drug introduced while preparing microspheres+Amount of polymer introduced while preparing microspheres)×100%
Measured drug entrapment efficiency (%) (A)=Measured amount (mg) of drug actually included in every 100 mg of microspheres immediately after preparation in comparative examples and examples/(100 mg of microspheres×theoretical drug loading amount (%))×100%
Drug entrapment efficiency after washing with ethanol (%) (B)=Measured amount (mg) of drug actually included in every 100 mg of microspheres after washing in experimental example 2/(100 mg of microspheres×theoretical drug loading amount (%))×100%
As can be seen in comparative example 1 and comparative example 1-1, the amount of drug crystals present outside exceeded 2% in the microspheres prepared without applying a process of additional washing with ethanol or a surfactant.
However, in the case of the examples prepared by adding a salt to the continuous phase, the amount of drug crystals confirmed to be present outside the microspheres was very small.
Therefore, when microspheres are prepared by adding a salt such as NaCl to the continuous phase, the amount of drug crystals that may be present outside the microspheres may be minimized, and thus the actual entrapment efficiency of the drug in the microspheres may be maximized.
10 mg of the microspheres prepared in the examples and comparative examples were respectively added to 100 mL of a pH 7.4 HEPES solution and placed in a constant temperature shaking water bath maintained at 37.0° C. After 24 hours, the supernatant was taken and filtered with a 0.45 μm syringe filter.
The initial daily dissolution amount of donepezil released from the microspheres was measured using HPLC. The column was XTerra Shield RP18 column 5 μm, 4.6×150 mm, the injection amount was 20 μl, the detection wavelength was 271 nm, and the mobile phase was pH 5.0 phosphate buffer solution and an acetonitrile solution (phosphate buffer solution:acetonitrile=60:40).
Table 2 below shows the dissolution rate for the 1st day of donepezil microspheres according to each example and comparative example.
As can be seen in Table 2, example 1 (where NaCl was added in 1% to the continuous phase) and example 2 (where NaCl was added in 5% to the continuous phase) showed a relatively lower dissolution rate compared to comparative example 1 (where NaCl was not added to the continuous phase).
As in experimental example 3, the smaller the amount of drug crystals present outside the microspheres, the lower the dissolution rate of the 1st day.
As can be seen in example 3 and example 3-1, when KCl was used, it was confirmed that the same effect as that of the case of NaCl was exhibited.
In addition, even when the polymer in dispersed phase was PLGA (Resomer RG 753 H), the initial dissolution rate of microspheres in example 4 was lower than the initial dissolution rate of comparative example 3.
In order to confirm the effect of suppressing the initial release of microspheres prepared by adding salt to the continuous phase, the concentration of donepezil in the blood was measured after subcutaneous administration to the back of the neck of rats.
The microspheres prepared in example 1, comparative example 1, comparative example 2, and comparative example 2-1 were weighed so that the amount of donepezil administered in the microspheres per rat was 25.2 mg/kg, and then dispersed in 0.3 mL suspension and subcutaneously injected into SD rats.
At regular intervals, 0.3 mL of blood was collected from the jugular vein of the rat, kept in an ice-cooled state, and centrifuged to separate 100 uL of plasma. The separated plasma was analyzed for the concentration of donepezil using LC/MS/MS.
The measurement results are shown in
As can be seen in
The microspheres of Example 1 prepared by adding NaCl in a content of 1% to the continuous phase showed the lowest Cmax of 58.9 ng/mL because drug crystals around the microspheres were removed and surface pores were also removed. In other words, it was possible to prevent the release of an excessive amount of drug in the initial stage. This result was similar to the in vitro result.
When microspheres are prepared by adding NaCl to the continuous phase according to the present invention, the formation of drug crystals around the microspheres is suppressed, and thus a separate process of removing drug crystals may not be additionally introduced and also the drug entrapment efficiency of the microspheres may be improved. In addition, drug crystals are not formed around the microspheres, and thus initial drug release may be suppressed when the microspheres are injected into the body.
According to the present invention, drug crystals are not formed around the microspheres, and thus initial drug release may be suppressed when the microspheres are injected into the body. In addition, the drug is released continuously, and thus the medicinal effect may be exhibited for a fairly long time with a single injection.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2019-0142635 | Nov 2019 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2020/015565 | 11/8/2020 | WO |
