Preparation and evaluation of oxaliplatin-loaded multivesicular liposome for early post-operative intraperitoneal chemotherapy

이주승 Graduate School, Yonsei University 2021 국내석사

Peritoneal carcinomatosis (PC) occurs in 10% of colorectal cancer patients, the third largest cancer patient in the world, with a poor prognosis. Colorectal cancer (CRC) with peritoneal metastasis is treated with intraperitoneal chemotherapy after tumor reduction surgery. In this study, oxaliplatin-loaded MVL (OX MVL) for EPIC regimen, one of intraperitoneal chemotherapy, was prepared by varying various lipid compositions (phospholipids, cholesterol, triolein) to improve particle size, %EE, morphological characteristics, and in vitro release and the optimized formulation was selected. For the selected depot, cytotoxicity test was performed on the HCT116 human colon cancer cell line of blank MVL, OX MVL, and 5% DEX. It was confirmed that blank MVL was not toxic at a concentration of 0.01 to 1 mg/mL, and OX MVL was less toxic than OX 5% DEX at the initial 24 hours, but the toxicity was the same after the last 72 hours. Biodistribution in BALB/c mice was conducted with IVIS equipment. It was observed that MVL remained in the liver and spleen for more than 5 days, while 5% DEX was excreted after 1 hour in the liver and spleen. In in vivo anticancer effect, a model that metastasized only to the abdominal wall (IPS) and a peritoneal viii metastasis cancer model (IPC) were established using CT26 in BALB/c mice. In model IPS, it was observed that the mean survival time of OX MVL was longer than that of OX 5% DEX at the same concentration. (log-rank test, p-value <0.001) In model IPC, the mean survival time of OX MVL was 17.92±1.72 days, which was the highest group. A lower survival rate was observed for IPS treated groups due to the higher dose of oxaliplatin although control groups survived well. IPC groups were further treated with adjusted doses increased survival rate than both control and solution treated groups. Therefore, MVL-loaded oxaliplatin could be a new regimen for CRC treatment in the IPC model.

Donepezil ionic liquid for enhancing solubility and skin permeability

현상민 Graduate School, Yonsei University 2019 국내박사

도네페질은 경구투여시 위장관의 부작용 및 치매환자 특유의 낮은 복약순응도 문제를 가지고 있다. 피부 투과도가 좋지않은 도네페질은 투과 촉진제를 사용하는 경우가 많은데 이는 인체 피부에 독성 또는 자극성을 유발할 수 있다. 이 연구의 목적은 도네페질 이온성 액체를 개발하고 경피 투여 가능한 효과적인 서방성 도네페질 패치 제제를 제형화하는 데 있다. 이온성 액체 스크리닝 연구를 위해, 다양한 코포머와 도네페질 혼합물을 30분간 유발 유봉으로 연마하였고 성상에 변화가 있는 DNP-ADI, DNP-ML, DNP-MA, DNP-SUC, DNP-TAR, DNP-VN의 6 가지 샘플이 관찰되었다. DSC 결과에 따르면 이온성 액체의 Tg는 DNP-VN을 제외하고는 실온 이하였다. 연한 노란색의 투명한 DNP-MA 혼합물은 IL 혼합물 중 -24.38 ℃의 가장 낮은 Tg 값과 1509µg/cm2의 가장 높은 피부 투과도를 가지고 있다. DNP-MA IL의 최적 비율을 찾기 위해 두 가지 물질을 다양한 몰비로 에탄올에 섞어 용매증발법으로 시료를 제조하였다. DNP-MA 이온성 액체 형성의 최적 몰비는 Tg 값 및 현미경 측정결과를 바탕으로 F4, 즉 0.2 : 0.8 몰비에 가까운 것으로 예측되었다. 또한 말론산의 함량이 증가할수록 피부 투과성이 증가하여 말론산이 피부 투과도를 향상시킬 수 있음을 나타냈다. 패치의 투명도, 투과성 및 피부 자극을 비교하기 위해 말론산 농도가 다른 DNP-MA IL 패치 (0 %, 1 %, 5 %, 10 %) 및 5%의 말산, 숙신산, 주석산 IL 패치를 준비했다. DNP 이온성 액체는 투명한 성상을 가지기 때문에 패치에 적용시 결정 형성을 억제하는 효과를 나타낸다. 패치 성상을 비교한 결과, 다량의 말론산을 사용할수록 결정 형성을 억제하는데 더 효과적이라는 것이 시각적으로 확인되었다. DNP-MA IL 패치 및 raw DNP 패치는 투과도에서도 큰 차이를 보였다. DNP-MA IL 패치에 대한 피부 투과 계수는 168시간동안 1.15 μg / cm2 / hr이였으며, raw DNP 패치보다 약 6 배 더 높았다. 따라서 본 연구의 DNP IL 패치는 결정생성 억제효과와 개선된 피부투과도를 나타낸 결과를 보였다. 이를 통해 기존 경구제형에 비해 더 나은 사용 편의성과 서방형 약물전달 효과를 기반으로 개선된 환자 복약순응도를 갖는 경피약물전달 시스템으로 적용될 수 있을 것이다. Orally administered DNP has the side effects of GI tract and compliances problem of dementia patients. DNP has poor skin permeation, transdermal formulation of DNP is not available except for permeation enhancer which have toxicity or irritation to human skin. The aim of this study was to investigate the DNP ionic liquid and formulate transdermally effective sustained release donepezil patch formulation. For the ionic liquid screening study, DNP mixtures with various coformers were ground with mortar and pestle for 30 min. 6 samples with visual changes were observed, those are DNP-ADI, DNP-ML, DNP-MA, DNP-SUC, DNP-TAR, DNP-VN. DSC results showed that Tg of the ionic liquids were below room temperature except for DNP-VN. The pale yellow transparent DNP-MA mixture has the lowest Tg value of -24.38℃ and the highest skin permeability of 1509µg/cm2 at 12 hours among IL mixtures. For investigating the optimal ratio of DNP-MA IL, samples with various molar ratio of two materials were prepared by solvent evaporation method. The optimum molar ratio of DNP-MA ionic liquid formation was predicted to be near F4, 0.2:0.8 molar ratio, by Tg and morphology with temperature control. Moreover, as the content of MA increased, the skin permeability increased, indicating that MA is capable of enhancing skin permeation. DNP IL patches with different coformers (ML, SUC, TAR) and MA concentrations (0%, 1%, 5%, 10%) were prepared to compare transparency. It exhibits the effect of suppressing crystal formation in the patch because DNP ionic liquid has an advantage of transparent appearance. As a result of confirming the appearance and morphology, patches are confirmed that the more malonic acid is used, the more effective to inhibit crystal formation. DNP-MA IL patches and the raw DNP patch showed a big difference in permeability. The skin permeability coefficients for 168 hours of DNP-MA IL patch is 1.15 µg/cm2/hr, 6 times higher than that of DNP raw patch which is 0.19 µg/cm2/hr. This study demonstrates that the DNP IL patch achieved the inhibition effect of crystal formation and enhanced solubility and permeability. This could be applied to further transdermal drug delivery system with improved patient’s compliance and sustained release.

Process optimization of spray drying of lysozyme loaded liposomes fabricated by microfluidics

Khraisat, Rama Graduate School, Yonsei University 2023 국내석사

흡입용 건조 분말(Dry powder for inhalation, DPI)은 천식과 당뇨병을 비롯한 지역 및 전신 질환 치료를 위해 단백질과 펩타이드를 폐로 전달하기 위해 조사되고 있다. 단백질과 펩타이드는 생리화학적으로 불안정하여 제조, 저장 및 투여 중 단백질의 변성을 방지하기 위해 리포좀과 같은 운반체가 필요하다. 리포좀 형성물을 건조시키는 방법 중 하나는 스프레이 건조이다. 그러나 불안정한 단백질과 펩타이드의 무결성을 보호하고, 흡입 가능한 입자 크기의 고수율 분말을 생산하기 위해서는 스프레이 건조 매개변수를 최적화해야 한다. 이 연구에서는 모델 약물로 리소자임을 사용했습니다. Dolomite 5 입력 칩 3D 에 연결된 프로그램 가능한 단일 주사 마이크로플루이딕스 펌프를 사용하여 리소자임을 함유한 리포좀을 제조했다. 입자 크기, 다변량분산지수, 제타 포텐셜과 같은 리포좀 형성물의 물리화학적 특성을 평가하여 스프레이 건조의 최적 조성을 결정하기 위해 실험계획법(Design of Experiment, DOE)을 사용했다. 생산 과정에서 유속 비율 (Flow Rate Ratio, FRR)을 증가시키면 리소자임의 포획 효율이 향상되는 것을 관찰할 수 있었다. 통계 분석 결과, 분무 압력과 로이신 함량이 스프레이 건조 과정에 영향을 미치는 중요한 요인임을 확인했다. 최적화된 스프레이 건조 분말 조성은 단백질 기반 치료제의 효율적인 흡입 전달을 위한 유망한 가능성을 갖고 있다. Dry powder for inhalation (DPI) has been under investigation for the delivery of proteins and peptides to the lung to treat local and systemic diseases including asthma and diabetes. Proteins and peptides suffer from physiochemical instability emerge the need for a carrier like liposomes to protect them from denaturation during manufacturing, storage, and administration. One of the methods used to dry liposomal formulations is spray drying. However, spray drying parameters needed to be optimized to protect the unstable protein and peptides integrity, and produce powder with high yield inhalable particle size. In this study, lysozyme was used as a model drug. Lysozyme-loaded liposomes were prepared using programmable single-syringe microfluidics pump connected to Dolomite 5 input chip 3D. The physicochemical characteristics of the liposomal formulations, such as particle size, polydispersity index, and zeta potential, were evaluated to determine the optimal formulation to be used in the optimization process of spray drying using design of experiment approach (DOE). It was observed that increasing the flow rate ratio (FRR) during the production process improved the encapsulation efficiency of lysozymes. Statistical analysis revealed that atomizing pressure and leucine content were significant factors influencing the spray drying process. The optimized spray-dried powder formulation holds promise for efficient delivery of proteins via inhalation, contributing to the development of effective protein-based therapeutics

Preparation and evaluation of protein-encapsulated liposome using poloxamer and sucrose ester surfactant for dry powder inhalation

박주현 Graduate School, Yonsei University 2023 국내석사

본 연구는 폴록사머188, 폴록사머 407그리고 자당지방산 에스테르 고체 계면활성제와 함께 제조하고 분무건조기기를 이용하여 흡입분말 리포좀 형태로 제조하고 그 영향을 평가하였다. 리포좀 이중층 구조를 구성하기 위하여 DSPC와 콜레스테롤을 사용하였고 라이소자임이 모델 단백질로 사용되었다. DSPC와 콜레스테롤의 적정 비율을 탐색한 후 고체 계면활성제의 종류에 따라 안정하고 균질한 형태의 리포좀을 제작하기 위해 고압균질기를 사용하여 제형을 제조하였다. 건조분말 형태의 파우더 리포좀을 입자사이즈, 제타전위, 라이소자임 정량, 봉입률, 잔류 수분량, 흐름성, 등을 측정하였고, 주사전자현미경(SEM)과 광학현미경으로 흡입분말로 제조된 리포좀의 형태를 확인하였다. 자당지방산 에스테르 S-1670을 고형 계면활성제로서 사용한 제형이 라이소자임 정량을 통해 흡입분말 리포좀에 봉입된 라이소자임의 봉입률은 29.21%이다. 최종적으로 in vitro 에어로졸화 효능 평가를 통한 방출량(EF%) 결과는 93%이며, 미세입자분획 (FPF%) 결과가 24.6%로 미세한 입자로서 수득 되었다. 이는 폐 깊은 곳까지 제형이 도달할 수 있다는 것을 나타내며, 고형 계면활성제의 종류에 따라 수득 되는 제형을 확인하고 흡입 분말 리포좀에 사용한 고형 계면활성제의 제조 및 평가 연구의 가능성을 보여주었다. In this study, poloxamer 188, poloxamer 407, and sucrose fatty acid ester were used as solid surfactant in the preparation of inhaled powder liposomes a spray drying machine, and the effect was evaluated. DSPC, and cholesterol were used to construct the liposomal bilayer structure, and lysozyme was used as a model protein. After exploring the appropriate ratio of DSPC, and cholesterol, formulations were prepared using a high-pressure homogenizer to prepare stable, and homogeneous liposomes according to the type of solid surfactant. Powder liposomes in the form of dry powder were characterized after measuring particle size, zeta potential, lysozyme quantity, encapsulation efficiency (EE%), residual moisture content, flowability, and morphology was observed. The lysozyme encapsulation rate of the formulation using sucrose fatty acid ester s-1670 as a solid surfactant was 29.21% through lysozyme quantification. Finally, the emission fraction (EF%) results was 93%, and the fine particle fraction (FPF%) result was 24.6%, which means the fine particles could reach deep into the lungs, through in vitro aerosolization efficacy evaluation. The formulation obtained according to the above was confirmed, and the possibility of manufacturing, and evaluating solid surfactants used for inhalation powder liposomes was demonstrated.

Desing and evaluation of donepezil ionic liquid patch

이수헌 Graduate School, Yonsei University 2020 국내석사

본 연구는 도네페질 이온성 액체를 제조 및 평가, 아크릴계 점착제를 이용하여 패취제조시 피부투과도를 향상 시키고 안정성을 향상 시켜 결정생성을 억제하는 경피패취제 제조와 비교 평가에 그 목적이 있다. 도네페질은 아세틸콜린에스테라제 억제제로서 알츠하이머병 치료에 사용된다. 현재 상용되고 있는 도네페질 제제는 정제로 경구투여가 이루어진다. 하지만, 경구투여시 위장관 부작용과 낮은 복약순응도로 인해 문제가 야기된다. 본 연구에서는 도네페질과 코포머로서 디카르복실산을 에탄올에 녹여 혼합 후 건조를 통해 이온성 액체를 제조하였다. 이 이온성 액체는 도네페질과 디카르복실산이 이온결합하여 상온에서 안정한 액체성상을 유지하는 것을 확인하였다. 이온성액체를 규명하기 위한 실험으로 DSC, Optical microscope 실험을 진행하였고 향상된 피부투과도를 확인하기 위하여 in-vitro 피부투과도 실험을 진행하였다. DSC결과에 따르면 도네페질 이온성 액체는 -30℃ ~ -10℃ 사이에서 유리전이온도를 가지는 것을 확인할 수 있으며 이는 상온에서 액체 및 유동성 페이스트 상태임을 의미한다. 이를 보완하기 위하여 광학현미경을 통하여 상온에서 무정형 상태를 유지하는 것을 확인하였다. in-vitro 피부투과도 평가를 통하여 도네페질 이온성 액체가 도네페질 보다 우수한 피부투과도를 보이는 것을 확인하였다. 패취제 제조시 가장 널리 이용되는 헨켈사의 아크릴계 점착제를 이용하여 패취제를 제조하고 이의 피부투과도를 평가하였다. Durotak 87-2051, Durotak 87-2074, Durotak 87-2196 이 3가지 아크릴계 점착제를 점착제로 사용하고 PEG400, 과 Tween80을 첨가제로 사용하여 패취를 제조한 뒤 Ex-vivo 피부투과도 평가 실험을 진행하였다. 연구결과를 통해 도네페질 이온성 액체는 패취제의 결정생성과 투과도를 향상시킴으로써 도네페질을 함유한 경피흡수제제 개발의 훌륭한 제제 기술로 사려된다. The purpose of this study is to prepare and evaluate donepezil ionic liquid patch. Specifically, it aims to produce a patch that inhibits crystal formation and improves skin permeability. Donepezil is an acetylcholinesterase inhibitor used to treat Alzheimer's disease. Currently, donepezil products are administered orally in tablets. However, oral administration causes problems due to gastrointestinal side effects and low medication compliance. In this study, ionic liquids were prepared by dissolving dicarboxylic acid in ethanol with donepezil. It was confirmed that this ionic liquid was ionically bonded to donepezil and dicarboxylic acid to maintain a stable liquid phase at room temperature. DSC and optical microscope experiments were conducted to identify ionic liquids and in-vitro skin permeability experiments were performed to confirm the improved skin permeability. According to the DSC results, it can be seen that the donepezil ionic liquid has a glass transition temperature between -30 ° C and -10 ° C, which means that the liquid and fluid paste state at room temperature. Also, it was confirmed that the amorphous state was maintained at room temperature through an optical microscope. In-vitro skin permeability evaluation confirmed that the donepezil ionic liquid showed better skin permeability than donepezil. The patch was prepared using Henkel's acrylic adhesive, which is the most widely used patch preparation, and the skin permeability thereof was evaluated. Durotak 87-2051, Durotak 87-2074, and Durotak 87-2196 acrylic adhesives were used as adhesives, and PEG400, and Tween80 were used as additives to prepare patches. Ex-vivo skin permeability evaluation was conducted. In the final formulation, donepezil ionic liquid patches showed excellent skin permeability than the donepezil patch. Therefore, result from this study suggested that donepezil ionic liquid can inhibit crystallization and enhance skin permeability, so it can be considered excellent formulation technology for the development of transdermal patches.

Influence of plasticizers on the pharmaceutical properties of fenofibric acid microcapsules

이은영 Graduate School, Yonsei University 2023 국내석사

Pharmaceutical development of D-leucine methyl ester HCL tablet for sarcopenia

김건우 Graduate School, Yonsei University 2023 국내석사

In this study, a Quality by Design (QbD)-based pharmaceutical development was performed for D-Leucine methyl ester HCl, a new drug candidate for sarcopenia treatment, according to ICH Q8. As a result of the basic physical property test of the API, the flowability is very very poor and the critical relative humidity value is 60.06%, so there is a risk of strong humidity even in a general indoor environment. Therefore, the direct compression method and the dry granulation method have problems such as problems in tableting or improper molding. In this study, the wet granulation method and the moisture-proof coating were used to overcome these problems and to minimize the quality problem caused by the critical relative humidity. As the binder solution used for wet granulation confirmed the characteristics of API with high water solubility, ethanol with relatively low solubility was selected as a solvent, and PVP-K30 was used as a binder to improve tableting properties. The Quality Target Product Profile (QTPP) was established from the basic physical property data and the compatibility data, and the Critical Quality Attribute (CQA) was established accordingly, and the initial risk assessment of the formulation was conducted using the Failure Mode and Effects Analysis (FMEA) method, and the research was conducted in the direction of improving the risk to a 'low'. A simplex centroid design was conducted to select additives that could complement low flowability and high hygroscopicity and to select the optimal ratio of them. A ratio range suitable for an immediate-release formulation was found through a mixture design method, and naked tablets were compressed into tablets at a ratio within the range, and film-coated tablets were prepared by coating them. We succeeded in developing a formulation that satisfies the critical quality attributes of granules, naked tablets, and coated tablets and that satisfies all list of the risk assessment of the formulation in a ‘low’.

Preparation and Evaluation of Extended Release Pellets Containing Aspirin by the Fluid Bed Process

서상교 충남대학교 대학원 2011 국내석사

Design and evaluation of long-acting intratumoral injection of 5-Fluorouracil based on solvent exchange mechanism

김건주 Graduate School, Yonsei University 2020 국내석사

종양내 주사는 종양 내에 직접 약물을 주사하는 국소 항암 치료요법이다. 항암제의 전신적인 부작용을 피할 수 있고, 직접 암세포에 약물을 전달하기 때문에 더 낮은 약물용량으로 같거나 높은 치료효과를 기대할 수 있는 장점이 있다. 기존의 종양내 주사의 문제점으로는 주사 후 약물이 종양조직 밖으로 쉽게 유출된다는 점이 있다. 이에 본 연구에서 in situ gel 기술을 이용한 서방성 주사제형으로 제형을 설계하였다. 결장암, 직장암, 유방암, 간암 등 다양한 고형암치료에 사용되는 5-FU를 약물로 사용하였다. In situ gel을 구현하는 방식에는 온도감응성, pH감응성, 용매교환 등 다양한 기전이 존재한다. 본 연구는 용매교환을 통해 졸 겔 전이를 구현할 수 있는 기전을 이용하였다. 제형을 설계하는데 필요한 유기용매로는 NMP를 사용하였고, 고분자는 EUDRAGIT E, RS, RL 세 가지를 사용하였다. 각 제형의 졸 겔 전이를 확인하고, 겔 형성 능력 및 유지능력을 평가하였다. in vitro gel 분해 및 약물용출, 수분확산 시험, SD rat을 이용한 생분해성등을 평가하였다. Intratumoral injection is a local chemotherapy that injects drugs directly into a tumor. Systemic side effects of anticancer drugs can be avoided, and since drugs are delivered directly to cancer cells, the same or higher therapeutic effect can be expected at a lower drug dose. A problem of conventional intratumoral injection is that the drug easily flows out of the tumor tissue after the injection. In this study, we designed the formulation through solvent exchange mechanism. 5-FU, which is used to treat various solid cancers such as colon cancer, rectal cancer, breast cancer and liver cancer, was used as a drug. There are various mechanisms such as thermosensitivity, pH sensitivity, and solvent exchange in implementing in situ gel drug delivery system. This study used a mechanism to realize sol gel transition through solvent exchange. NMP was used as an organic solvent for designing the formulation, and three polymers were used as EUDRAGIT E, RS, and RL. The sol gel transition of each formulation was confirmed, and the gel formation ability and retention ability were evaluated. In vitro gel degradation, drug dissolution, water diffusion test, and biodegradability using SD rats were also evaluated.

Oxaliplatin loaded chemically cross-linking hydrogel composed of hyaluronic acid and carboxymethyl cellulose for preventing postoperative adhesion

Lee, JeeEun Graduate School, Yonsei University 2018 국내석사

Postoperative peritoneal adhesions are very common and cause severe complication after surgery. Biodegradable hydrogels derived from natural polysaccharides is ideal to prevent postoperative adhesion in peritoneum. Hyaluronic acid (HA) has the properties which are bioavailability and biodegradation and these properties make HA suitable for a various medical fields. However, HA have a limit to weak mechanical properties and rapid degradation. Carboxymethyl cellulose (CMC) is a highly water-soluble anionic polysaccharide and is used to prevent adhesion after surgery. This research is to synthesize a chemically cross-linked hydrogel composed of HA and CMCNa, thorough the used of adipic dihydrazide (ADH) as a cross linker and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) as a carboxyl activating agent with longer duration and oxaliplatin is dispersed in hydrogel to preparation anti-adhesion barrier with anticancer effect. Carboxyl group of HA and CMC was made cross-linked hydrogel by forming amide bond with ADH as a cross linker. The structure of cross-linked hydrogel is a porous structure as the ratio of HA increases and it can help the growth and proliferation of the cells. Cross-linked hydrogel has a psedoplastic behavior and it is related to the molecular weight of polymer. The viscosity increases when the ratio of HA increases. The dissolution rate of oxaliplatin dispersed in hydrogel was from 57 % to 65 % and it is related to the degradation of hydrogel. In case of the HA-CMCNa=1:1, 1:2, 2:1, these were showed similar degradation rate, and sustained for about 60 hours. In vivo experiment was carried out using the samples, which had a low degradation rate, HA-CMCNa=2:1 hydrogel showed high anti-adhesion effect.