KR20040066548A - Locally administrable, sustained-release microsphere for biological active peptide and their method of preparation - Google Patents
Locally administrable, sustained-release microsphere for biological active peptide and their method of preparation Download PDFInfo
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Abstract
Description
본 발명은 생리활성펩타이드의 국소투여용 서방성 마이크로스피어 및 그 제조방법에 관한 것이다.The present invention relates to sustained-release microspheres for topical administration of physiologically active peptides and methods for their preparation.
국소투여와 서방출을 목적으로 하는 생분해성 마이크로스피어의 제조에는 여러 방법들이 사용되고 있다. 적당한 제조 방법은 약물의 성질에 의존하는데, 마이크로스피어의 형태(morphology)와 마이크로스피어의 약물 방출 양상에 커다란 영향을 줄 수 있다.Various methods have been used to prepare biodegradable microspheres for topical administration and slow release. The proper method of preparation depends on the nature of the drug, which can have a significant impact on the morphology of the microspheres and the pattern of drug release of the microspheres.
일본특허공보 제33414호에는 지용성 약물과 생분해성 고분자를 유기용매에 용해시키고 상분리제를 가하여 에멀젼을 형성시킨 후 용매를 증발시켜 마이크로스피어를 제조하는 용매증발법이 개시되어 있다.Japanese Patent No. 33414 discloses a solvent evaporation method in which a fat-soluble drug and a biodegradable polymer are dissolved in an organic solvent, a phase separation agent is added to form an emulsion, and then the solvent is evaporated to produce microspheres.
보다 개선된 방법으로, 일본특허공보 제100516호에는 수용성 약물과 생분해성 고분자를 이용하여 w/o/w 다중유제법(water in oil in water)에 의하여 마이크로스피어를 제조하는 방법이 개시되어 있다. 이 방법은 젤라틴(gelatin)의 점도를 이용하여 수용성 펩타이드의 봉입효율을 개선한 것이다. 그러나, 젤라틴은 동물 콜라겐 단백질에 산 또는 염기를 처리하여 얻은 것으로, 젤라틴의 점도가 온도에 민감하기 때문에 각 제조시 마다 편차가 발생할 수 있다는 문제점이 있다.As a further improved method, Japanese Patent Publication No. 100516 discloses a method for preparing microspheres by w / o / w water in oil in water using water-soluble drugs and biodegradable polymers. This method uses the viscosity of gelatin to improve the encapsulation efficiency of water-soluble peptides. However, gelatin is obtained by treating an animal collagen protein with an acid or a base, and since the viscosity of the gelatin is sensitive to temperature, there is a problem that a deviation may occur in each preparation.
한편, 알긴산염은 1β→4-D-만누론산(mannuronic acid)과 α-L-글루쿠론산(glucuronic acid)의 음이온성 공중합체로, 물에 녹이면 점성이 큰 용액이 되므로, 식품 첨가제, 정제붕해제, 겔화제 등으로 사용되어 왔다. 또한, 알긴산염은 독성이 없고 생체내에서 분해되므로, 세포를 고정화하거나, 약물 또는 항원의 전달시스템으로도 광범위하게 이용되어 왔다.Alginate, on the other hand, is an anionic copolymer of 1β → 4-D-mannuronic acid and α-L-glucuronic acid.It is a viscous solution when dissolved in water. It has been used as a disintegrant, a gelling agent and the like. In addition, since alginate is non-toxic and degraded in vivo, it has been widely used for immobilizing cells or as a delivery system for drugs or antigens.
알긴산염의 가장 큰 특징 중의 하나는 칼슘 이온과 같은 2가 양이온의 존재하에 겔을 형성한다는 것으로, 이러한 알긴산염 겔은 수용액에서 제조될 수 있고 생체내에서 팽윤하여 점차적으로 붕해되기 때문에 마이크로스피어의 고분자 기질로서 많이 이용되는 이유가 된다. 그러나, 알긴산염 마이크로스피어가 종래에 약물의 수송체로서 이용되어 왔다 할지라도, 약물 방출 속도를 결정하기 위한 최적의 물리, 화학적 조건들이 정립되어 있는 것은 아니므로, 알긴산염의 특성을 이용하여 원하는 방출특성을 가진 안정적인 마이크로스피어를 얻는 것은 아직도 많은 연구가 필요한 분야이다.One of the most distinctive features of alginates is the formation of gels in the presence of divalent cations such as calcium ions. These alginate gels can be prepared in aqueous solutions and swell in vivo and gradually disintegrate, thus resulting in the polymeric matrix of microspheres. As a reason, it is used a lot. However, although alginate microspheres have conventionally been used as drug carriers, optimal physical and chemical conditions for determining drug release rates have not been established. Acquiring stable microspheres with high resolution is still a subject of much research.
본 발명에서는 국소투여와 서방출을 목적으로 하는 생분해성 마이크로스피어의 제조시에 나타나는 상기와 같은 문제점을 해결하기 위하여, 최대한 제조 편차가 발생하지 않도록 점도의 영향 없이 마이크로스피어를 만들고자 하며, 또한 약물, 특히 고가의 생리활성펩타이드 약물의 봉입효율을 높이는 효율적인 방법으로 마이크로스피어를 만들고자 한다. 본 발명은 이와 같이, 제조편차 없이 약물의 손실을 최소화하는 방법으로 원하는 기간 동안 지속적으로 방출되는 생리활성펩타이드의 서방성 마이크로스피어를 제조하는 것을 목적으로 한다.In the present invention, in order to solve the above problems appearing in the production of biodegradable microspheres for the purpose of topical administration and sustained release, to make a microsphere without the effect of viscosity so as not to produce a deviation as much as possible, In particular, it is intended to make microspheres in an efficient way to increase the encapsulation efficiency of expensive bioactive peptide drugs. As described above, an object of the present invention is to prepare sustained-release microspheres of bioactive peptides that are continuously released for a desired period of time in a manner of minimizing drug loss without preparation deviation.
도 1은 용출율 실험 결과를 그래프로 나타낸 것이다.Figure 1 shows the dissolution rate experiment results graphically.
상기와 같은 목적을 달성하기 위하여 본 발명에서는,In order to achieve the above object, in the present invention,
(a) 생리활성펩타이드 약물 1 중량부와 알긴산염 0.001∼0.1 중량부를 정제수에 용해시켜 수상의 용해액(A)을 만들고,(a) 1 part by weight of the bioactive peptide drug and 0.001 to 0.1 parts by weight of alginate are dissolved in purified water to form an aqueous solution (A).
(b) 폴리락타이드(Polylactide, PLA); 폴리글리콜라이드(Polyglycolide, PGA); 폴리(락타이드-코-글리콜라이드)(Poly(lactide-co-glycolide): PLGA) 중에서 선택된 1종 이상의 생분해성 고분자 1∼20 중량부를 유기용매에 용해시켜 유상의 용해액(B)을 만든 후 이 유상의 용해액(B)에 상기 수상의 용해액(A)를 분산시켜 1차 에멀젼(C)을 제조하고,(b) Polylactide (PLA); Polyglycolide (PGA); 1 to 20 parts by weight of one or more biodegradable polymers selected from poly (lactide-co-glycolide) (PLGA) are dissolved in an organic solvent to form an oily solution (B). The primary emulsion (C) is prepared by dispersing the aqueous solution (A) in the aqueous phase in the oily solution (B).
(c) 양이온성염 10∼30 중량부를 수성용제에 용해시켜 수용액(D)을 만든 후 이 수용액상(D)에 상기 1차 에멀젼(C)을 분산시켜 표면겔화기법으로 제조되는 것을 특징으로 하는 생리활성펩타이드의 서방성 마이크로스피어와 그 제조방법 및 이를포함하는 제제가 제공된다.(c) 10-30 parts by weight of cationic salts are dissolved in an aqueous solvent to form an aqueous solution (D), and then the primary emulsion (C) is dispersed in this aqueous phase (D) to be prepared by a surface gelation method. Sustained release microspheres of an active peptide, methods for preparing the same, and preparations containing the same are provided.
본 발명에서 마이크로스피어로 제조하는 약물은, 생리활성펩타이드를 목적으로 하나 이밖에 단백질 약물, 국소마취제, 항암제, 진정제, 간질 치료제, 항궤양제, 항우울증제, 항알레르기제, 강심제, 항부정맥제, 혈관확장제, 저혈압성 이뇨제, 당뇨병 치료제, 과지질혈증 치료제, 항응고제, 용혈제, 항결핵제, 호르몬, 마취 길항제, 골흡수 억제제, 골형성 촉진제 또는 혈관형성 억제제 등과 이들 약물들의 염형태도 적절한 조건하에 본 발명의 방법에 따라 국소투여용 서방성 마이크로스피어로 제조될 수 있다.In the present invention, the drug prepared by microspheres, for the purpose of bioactive peptides, protein drugs, local anesthetics, anticancers, sedatives, epilepsy treatments, anti-ulcers, antidepressants, antiallergic agents, cardiovascular agents, antiarrhythmic agents, Vasodilators, hypotensive diuretics, diabetes treatments, hyperlipidemia treatments, anticoagulants, hemolytics, anti-tuberculosis agents, hormones, anesthetic antagonists, bone resorption inhibitors, bone formation promoters or angiogenesis inhibitors, etc. Depending on the method, it may be prepared as a sustained release microsphere for topical administration.
본 발명의 생리활성펩타이드 약물은, 특히 초산루프로렐린(Leuprorelin acetate), 초산부세렐린(Buserelin acetate), 초산고세렐린(Goserelin acetate) 및 초산트립토렐린(Triptorelin acetate) 중 하나를 선택적으로 포함한다.The bioactive peptide drug of the present invention, in particular, optionally comprises one of Leuprorelin acetate, Buserelin acetate, Goserelin acetate and Triptorelin acetate .
본 발명에 사용되는 양이온성염은 염화칼슘, 염화바륨, 황산알루미늄, 황산아연, 황산구리, 질산은, 염화제이철 등의 2가 금속염이다.Cationic salts used in the present invention are divalent metal salts such as calcium chloride, barium chloride, aluminum sulfate, zinc sulfate, copper sulfate, silver nitrate and ferric chloride.
본 발명에서 생분해성 고분자를 용해시키는 유기용매로는, 특히 한정되는 것은 아니나, 일반적으로 염화메틸렌이 사용될 수 있다.In the present invention, the organic solvent for dissolving the biodegradable polymer is not particularly limited, but methylene chloride may be generally used.
본 발명에서 양이온성염을 용해시키는 수성용제로는, 양이온성염을 용해시킬 수 있고 의약품의 제조에 사용될 수 있는 수성용제면 제한없이 사용할 수 있으며, 일예로 폴리비닐알코올 등이 사용될 수 있다.As the aqueous solvent for dissolving the cationic salt in the present invention, it can be used without limitation of the aqueous solvent which can dissolve the cationic salt and can be used in the manufacture of pharmaceuticals, for example polyvinyl alcohol and the like can be used.
본 발명의 마이크로스피어는, 약제학적으로 허용 가능한 통상의 부형제나 보조제 등을 첨가, 혼합하여 통상의 약제학적 방법으로 제제화할 수 있으며, 바람직하게는 통상의 부형제나 보조제를 참가하여 피하주사용 서방성 제제의 형태로 제제화할 수 있다.The microspheres of the present invention can be formulated in a conventional pharmaceutical method by adding and mixing pharmaceutically acceptable conventional excipients, adjuvants and the like, preferably subcutaneous sustained release for the use of conventional excipients and auxiliaries. It may be formulated in the form of a formulation.
본 발명에서 생리활성펩타이드의 서방성 마이크로스피어는 다음과 같은 단계를 포함하여 제조된다.Sustained release microspheres of the bioactive peptides in the present invention are prepared including the following steps.
(1) 약물 1 중량부와 알긴산염 0.001∼0.1 중량부를 정제수에 용해시켜 수상(A)을 제조하는 단계와;(1) dissolving 1 part by weight of drug and 0.001 to 0.1 parts by weight of alginate in purified water to prepare a water phase (A);
(2) 생분해성 고분자 1∼20 중량부를 염화메칠렌에 녹여 유상을 제조하고, 전기과정에 의해 얻은 수상(A)을 유상(B)에 분산시켜 1차 에멀젼(C)을 얻는 단계;(2) dissolving 1 to 20 parts by weight of biodegradable polymer in methylene chloride to prepare an oil phase, and dispersing the water phase (A) obtained by the electrical process in the oil phase (B) to obtain a primary emulsion (C);
(3) 양이온성염 10∼30 중량부를 수성용제에 용해시켜 수상(D)을 만들고, 이 수상(D)에 전기과정에 의해 얻은 1차 에멀젼(C)을 분산시켜 표면겔화기법에 의해 마이크로스피어(E)를 제조하는 단계;(3) 10 to 30 parts by weight of the cationic salt is dissolved in an aqueous solvent to form an aqueous phase (D), and the primary emulsion (C) obtained by an electrical process is dispersed in this aqueous phase (D) to obtain a microsphere ( E) preparing;
(4) 전기과정에서 얻은 마이크로스피어(E)인 미립구를 회수하는 단계.(4) recovering microspheres which are microspheres (E) obtained in the previous process.
본 발명에 따라 제조된 마이크로스피어에서, 알긴산염은 약물과 함께 수상(A)에 포함되고 1차 에멀젼 제조시 유상(B)에 둘러 쌓이게 된다. 통상은 이 1차 에멀젼(C)을 다시 수상에 분산시킬 때 알긴산염과 약물이 수상에 노출되어 약물의 손실이 발생하게 된다. 그러나, 본 발명에서는 수상에 양이온성 염을 첨가함으로써 알긴산염이 수상(D)에 노출시 표면에서 단단한 겔을 형성하게 되어 약물 손실을 최소화할 수 있다. 이는 알긴산염이 2가 이상의 금속이온을 가하면 즉시 겔을 형성하고 이러한 겔 형성은 주로 이온과 동종 공중합체 블럭 사이의 결합점에서 주로 일어나며 이온결합에 의한 가교 형성으로 단단한 겔을 만들 수 있다는 특징을 이용한 것으로, 본 발명에서 알긴산염은 분자량 및 점도에 관계없이 사용될 수 있다.In the microspheres prepared according to the invention, the alginate is included in the aqueous phase (A) together with the drug and enclosed in the oil phase (B) during the preparation of the primary emulsion. Normally, when the primary emulsion (C) is dispersed again in the aqueous phase, the alginate and the drug are exposed to the aqueous phase, resulting in the loss of the drug. However, in the present invention, by adding a cationic salt to the aqueous phase, the alginate forms a hard gel on the surface when exposed to the aqueous phase (D), thereby minimizing drug loss. This is due to the fact that alginate forms a gel immediately upon addition of a divalent or higher metal ion, and this gel formation occurs mainly at the point of attachment between the ions and homopolymer blocks, and can be used to form a hard gel by crosslinking by ionic bonding. In the present invention, alginate may be used regardless of molecular weight and viscosity.
본 발명에 따라 제조된 마이크로스피어를 포함하는 서방성 제제는 약물 방출기간이 10일 내지 4개월 범위에서 조절될 수 있다.Sustained release formulations comprising the microspheres prepared according to the present invention may have a drug release period of 10 days to 4 months.
이하, 실시예 및 실험예에 의해 본 발명을 보다 상세하게 설명하고자 한다. 그러나, 다음의 실시예 및 실험예에 본 발명의 기술적 범위가 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the technical scope of the present invention is not limited to the following Examples and Experimental Examples.
실시예 1∼4Examples 1-4
초산루프로렐린 마이크로스피어의 제조Preparation of Luproline Acetate Microspheres
다음의 표 1과 같은 조성으로 본 발명에 따른 마이크로스피어를 제조하였다.To prepare a microsphere according to the invention in the composition shown in Table 1.
* RESOMER RG502H : Poly(D,L-Lactide-co-glycolide) 50:50, M.W. 8,000, 베링거인겔하임RESOMER RG502H: Poly (D, L-Lactide-co-glycolide) 50:50, M.W. 8,000, Boehringer Ingelheim
* RESOMER RG503H : Poly(D,L-Lactide-co-glycolide) 50:50, M.W. 33000, 베링거인겔하임RESOMER RG503H: Poly (D, L-Lactide-co-glycolide) 50:50, M.W. 33000, Boehringer Ingelheim
* RESOMER RG502 : Poly(D,L-Lactide-co-glycolide) 50:50, M.W. 14,500, 베링거인겔하임RESOMER RG502: Poly (D, L-Lactide-co-glycolide) 50:50, M.W. 14,500 Boehringer Ingelheim
* RESOMER RG752 : Poly(D,L-Lactide-co-glycolide) 75:25, M.W. 20,000, 베링거인겔하임RESOMER RG752: Poly (D, L-Lactide-co-glycolide) 75:25, M.W. 20,000, Boehringer Ingelheim
1) 초산루프로렐린 87.5mg과 알긴산나트륨 0.28mg을 정제수 140㎕에 용해시켜 수상의 용해액을 제조하고;1) A solution of aqueous phase was prepared by dissolving 87.5 mg of luproline acetate and 0.28 mg of sodium alginate in 140 µl of purified water;
2) 생분해성 고분자 750mg을 염화메칠렌 2mL에 녹여 유상의 용해액을 제조하고;2) dissolving 750 mg of biodegradable polymer in 2 mL of methylene chloride to prepare an oily solution;
3) 1)의 용해액과 2)의 용해액을 혼합하고 분산시켜 1차 에멀젼을 제조하고;3) the primary emulsion was prepared by mixing and dispersing the solution of 1) and the solution of 2);
4) 염화칼슘 2.5g을 0.2% 폴리비닐알코올 용액 250mL에 용해시켜 수상의 용해액을 제조하고;4) 2.5 g of calcium chloride was dissolved in 250 mL of a 0.2% polyvinyl alcohol solution to prepare a solution of aqueous phase;
5) 3)에서 얻은 1차 에멀젼을 4)의 용해액에 분산시켜 표면겔화기법으로 마이크로스피어를 제조하고;5) dispersing the primary emulsion obtained in 3) in the solution of 4) to prepare a microsphere by the surface gelation method;
6) 5)의 마이크로스피어를 포함하는 용액을 0.45㎛ 멤브레인 필터를 통과시켜 여과하고 동결건조하여 마이크로스피어인 미립구를 회수하였다.6) The solution containing the microspheres of 5) was filtered through a 0.45 μm membrane filter and lyophilized to recover the microspheres as microspheres.
비교예 1∼2Comparative Examples 1 and 2
제조된 마이크로스피어의 효과를 비교 평가하기 위하여 다음의 표 2와 같은 조성으로 기존의 제조방법에 따라 마이크로스피어를 제조하였다. 비교예 1은 젤라틴을 사용하지 않고 표면겔화기법으로 제조하였으며, 비교예 1은 젤라틴을 사용하여 표면겔화기법으로 제조하였다.In order to compare and evaluate the effect of the prepared microspheres, the microspheres were prepared according to the conventional manufacturing method with the composition shown in Table 2 below. Comparative Example 1 was prepared by the surface gelation method without using gelatin, and Comparative Example 1 was prepared by the surface gelation method by using gelatin.
* RESOMER RG503H : Poly(D,L-Lactide-co-glycolide) 50:50, M.W. 33000, 베링거인겔하임RESOMER RG503H: Poly (D, L-Lactide-co-glycolide) 50:50, M.W. 33000, Boehringer Ingelheim
1) 비교예 1은 초산루프로렐린 87.5mg을, 비교예 2는 초산루프로렐린 87.5mg과 젤라틴 15mg을 정제수 140㎕에 용해시켜 수상의 용해액을 제조하고;1) Comparative Example 1 was prepared by dissolving 87.5 mg of luproline acetate, and Comparative Example 2 by dissolving 87.5 mg of luproline acetate and 15 mg of gelatin in 140 µl of purified water to prepare a solution of aqueous phase;
2) 생분해성 고분자 750mg을 염화메칠렌 2mL에 녹여 유상의 용해액을 제조하고;2) dissolving 750 mg of biodegradable polymer in 2 mL of methylene chloride to prepare an oily solution;
3) 1)의 용해액과 2)의 용해액을 혼합하고 분산시켜 1차 에멀젼을 제조하고;3) the primary emulsion was prepared by mixing and dispersing the solution of 1) and the solution of 2);
4) 염화칼슘 2.5g을 0.2% 폴리비닐알코올 용액 250mL에 용해시켜 수상의 용해액을 제조하고;4) 2.5 g of calcium chloride was dissolved in 250 mL of a 0.2% polyvinyl alcohol solution to prepare a solution of aqueous phase;
5) 3)에서 얻은 1차 에멀젼을 4)의 용해액에 분산시켜 표면겔화기법으로 마이크로스피어를 제조하고;5) dispersing the primary emulsion obtained in 3) in the solution of 4) to prepare a microsphere by the surface gelation method;
6) 5)의 마이크로스피어를 포함하는 용액을 0.45㎛ 멤브레인 필터를 통과시켜 여과하고 동결건조하여 마이크로스피어인 미립구를 회수하였다.6) The solution containing the microspheres of 5) was filtered through a 0.45 μm membrane filter and lyophilized to recover the microspheres as microspheres.
비교예 3Comparative Example 3
마이크로스피어 제조시 표면겔화기법의 효과를 비교평가하기 위하여 다음 표 3과 같은 조성으로 마이크로스피어를 제조하였다. 본 비교예에서는 알긴산염을 사용하나 양이온성염이 없어 알긴산염의 표면겔화가 발생하지 않는다.In order to compare and evaluate the effect of the surface gelation method in the preparation of the microspheres were prepared microspheres with the composition shown in Table 3. In this comparative example, alginate is used but there is no cationic salt, so that no surface gelation of the alginate occurs.
* RESOMER RG503H : Poly(D,L-Lactide-co-glycolide) 50:50, M.W. 33000, 베링거인겔하임RESOMER RG503H: Poly (D, L-Lactide-co-glycolide) 50:50, M.W. 33000, Boehringer Ingelheim
1) 초산루프로렐린 87.5mg과 알긴산나트륨 0.28mg을 정제수 140㎕에 용해시켜 수상의 용해액을 제조하고;1) A solution of aqueous phase was prepared by dissolving 87.5 mg of luproline acetate and 0.28 mg of sodium alginate in 140 µl of purified water;
2) 생분해성 고분자 750mg을 염화메칠렌 2mL에 녹여 유상의 용해액을 제조하고;2) dissolving 750 mg of biodegradable polymer in 2 mL of methylene chloride to prepare an oily solution;
3) 1)의 용해액과 2)의 용해액을 혼합하고 분산시켜 1차 에멀젼을 제조하고;3) the primary emulsion was prepared by mixing and dispersing the solution of 1) and the solution of 2);
4) 염화칼슘 2.5g을 0.2% 폴리비닐알코올 용액 250mL에 용해시켜 수상의 용해액을 제조하고;4) 2.5 g of calcium chloride was dissolved in 250 mL of a 0.2% polyvinyl alcohol solution to prepare a solution of aqueous phase;
5) 3)에서 얻은 1차 에멀젼을 4)의 용해액에 분산시켜 표면겔화기법으로 마이크로스피어를 제조하고;5) dispersing the primary emulsion obtained in 3) in the solution of 4) to prepare a microsphere by the surface gelation method;
6) 5)의 마이크로스피어를 포함하는 용액을 0.45㎛ 멤브레인 필터를 통과시켜 여과하고 동결건조하여 마이크로스피어인 미립구를 회수하였다.6) The solution containing the microspheres of 5) was filtered through a 0.45 μm membrane filter and lyophilized to recover the microspheres as microspheres.
실험예 1Experimental Example 1
봉입효율의 측정Measurement of encapsulation efficiency
실시예 1∼4 및 비교예 1∼3에서 제조한 마이크로스피어의 봉입효율을 측정하였다. 봉입효율은 이론함량에 대한 실제 함량의 비로서 제조시 약물의 손실을 나타낸다. 결과는 다음의 표 4와 같다.The sealing efficiency of the microspheres manufactured in Examples 1-4 and Comparative Examples 1-3 was measured. Encapsulation efficiency is the ratio of the actual content to the theoretical content, which represents the loss of the drug during manufacture. The results are shown in Table 4 below.
봉입효율이 높다는 것은 제조시 약물의 손실이 적다는 것으로, 본 발명의 방법으로 제조된 마이크로스피어(실시예 1∼4)는 92∼93%의 봉입효율을 나타낸 반면, 기존의 방법으로 제조된 비교예의 경우는 모두 78∼84%의 낮은 봉입효율을 나타내었다.High encapsulation efficiency means less loss of drug during preparation, whereas microspheres prepared by the method of the present invention (Examples 1-4) exhibited encapsulation efficiency of 92-93%, while comparisons made by conventional methods All of the examples showed low encapsulation efficiency of 78 to 84%.
실험예 2Experimental Example 2
in vitroin vitro 용출시험Dissolution Test
실시예 1∼4 및 비교실시예 1∼3에서 제조한 마이크로스피어의 in vitro 용출시험을 실시하였다. in vitro 용출시험은 마이크로스피어 7mg을 마개 달린 시험관에 넣고 소듐아자이드(sodium azide) 0.01%와 트윈(Tween)80 0.02%를 함유한 pH 7.4 PBS(Phosphate buffered saline) 용액 1.5㎖를 가하여 교반항온수조에서 실시하였다. 정해진 시간에 샘플를 원심분리하고 상징액을 제거한 후 동결건조하고, 고속액체크로마토그라프법을 이용하여 분석하였다. 결과는 다음의 표 5와 같으며, 이를 도 1에 그래프로 나타내었다.In vitro dissolution tests of the microspheres prepared in Examples 1 to 4 and Comparative Examples 1 to 3 were carried out. In vitro dissolution test was performed by adding 7 mg of microspheres to a test tube with a stopper and adding 1.5 ml of pH 7.4 Phosphate buffered saline (PBS) solution containing 0.01% sodium azide and 0.02% Tween80. It was carried out in. Samples were centrifuged at a defined time, the supernatant was removed and lyophilized, and analyzed using high performance liquid chromatography. The results are shown in Table 5 below, which is shown graphically in FIG. 1.
상기 실험 결과로부터 동일한 생분해성 고분자를 사용한 실시예 2와 비교예 1∼3의 용출율을 비교했을 때 실시예 2가 비교예 1∼3 보다 용출율이 2∼5% 정도 낮게 나오는 것을 확인할 수 있다. 본 발명의 방법으로 제조한 마이크로스피어의 경우 동일 폴리머를 사용한 비교예 1∼3보다 낮게 나온 것은 알긴산의 표면겔화기법의 영향이며, 또한 이를 이용하여 용출율을 조절할 수 있음을 보여주는 것이다.When the dissolution rate of Example 2 and Comparative Examples 1 to 3 using the same biodegradable polymer is compared from the above experimental results, it can be confirmed that the dissolution rate of Example 2 is about 2 to 5% lower than that of Comparative Examples 1 to 3. In the case of the microspheres prepared by the method of the present invention, lower than those of Comparative Examples 1 to 3 using the same polymer is an effect of the surface gelation method of alginic acid, and also shows that the dissolution rate can be controlled using the same.
본 발명의 마이크로스피어는 기존의 점도를 이용하여 제조된 방법에 비해 월등히 봉입효율이 높아 고가 약물의 손실이 없고 재현성이 높으며, 생분해성 고분자 각각의 농도 및 분자량 또는 양이온성 염의 농도를 조절함으로써 원하는 농도와 기간 동안 약효를 나타낼 수 있으므로, LHRH 동족체와 같은 생리활성펩타이드 약물을 필요 최소량으로 함유하여 장기간에 걸쳐 지속적으로 방출함으로써 치료효과를 극대화하고 부작용을 줄일 수 있다.The microspheres of the present invention have a significantly higher efficiency of encapsulation compared to the methods prepared using conventional viscosities, and thus have no loss of expensive drugs and have high reproducibility. Since the drug may be effective for a period of time, the drug may contain a minimum amount of a bioactive peptide drug such as LHRH homologue and continuously released over a long period of time, thereby maximizing a therapeutic effect and reducing side effects.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR100915601B1 (en) * | 2004-09-16 | 2009-09-07 | 베이징 홍이야오 사이언스 앤 테크놀로지 디벨롭먼트 컴퍼니 리미티드 | Vascular Embolus of Paclitaxel-Sodium Alginate Microsphere and its Prepartion |
| ES2358703A1 (en) * | 2007-11-14 | 2011-05-13 | Universidad Del Pais Vasco | Use of microparticles for use as vaccines and the liberation of biologically active molecules. (Machine-translation by Google Translate, not legally binding) |
| CN102988301A (en) * | 2012-12-26 | 2013-03-27 | 温州医学院 | Preparation method of long-acting sustained-release microspheres containing bevacizumab |
| WO2013094955A1 (en) * | 2011-12-19 | 2013-06-27 | 주식회사 삼양바이오팜 | Composition comprising biodegradable polymer fine particles having improved dispersibility and method for preparing same |
| WO2013119183A1 (en) * | 2012-02-06 | 2013-08-15 | Nanyang Technological University | Methods of manufacturing core-shell microparticles, and microparticles formed thereof |
| US20140356442A1 (en) * | 2011-12-22 | 2014-12-04 | Shangdong Luye Pharmaceutical Co., Ltd. | Pharmaceutical compositions of triptorelin microspheres |
| WO2021091333A1 (en) * | 2019-11-08 | 2021-05-14 | 환인제약 주식회사 | Microsphere for continuous release and method for manufacturing same |
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- 2003-01-20 KR KR1020030003642A patent/KR20040066548A/en not_active Application Discontinuation
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100915601B1 (en) * | 2004-09-16 | 2009-09-07 | 베이징 홍이야오 사이언스 앤 테크놀로지 디벨롭먼트 컴퍼니 리미티드 | Vascular Embolus of Paclitaxel-Sodium Alginate Microsphere and its Prepartion |
| ES2358703A1 (en) * | 2007-11-14 | 2011-05-13 | Universidad Del Pais Vasco | Use of microparticles for use as vaccines and the liberation of biologically active molecules. (Machine-translation by Google Translate, not legally binding) |
| WO2013094955A1 (en) * | 2011-12-19 | 2013-06-27 | 주식회사 삼양바이오팜 | Composition comprising biodegradable polymer fine particles having improved dispersibility and method for preparing same |
| US20140356442A1 (en) * | 2011-12-22 | 2014-12-04 | Shangdong Luye Pharmaceutical Co., Ltd. | Pharmaceutical compositions of triptorelin microspheres |
| WO2013119183A1 (en) * | 2012-02-06 | 2013-08-15 | Nanyang Technological University | Methods of manufacturing core-shell microparticles, and microparticles formed thereof |
| CN102988301A (en) * | 2012-12-26 | 2013-03-27 | 温州医学院 | Preparation method of long-acting sustained-release microspheres containing bevacizumab |
| WO2021091333A1 (en) * | 2019-11-08 | 2021-05-14 | 환인제약 주식회사 | Microsphere for continuous release and method for manufacturing same |
| CN114867470A (en) * | 2019-11-08 | 2022-08-05 | 丸仁制药株式会社 | Microspheres for sustained release and method of making same |
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