Oral Controlled Drug Delivery System of New Drug Candidate for Gastritis

장선우 성균관대학교 2006 국내박사

Gastroretentive controlled release delivery system of DA 6034, a new drug candidate for therapy of gastritis, was developed by using effervescent floating matrix system (EFMS). From the study of pharmacological efficacy with animal model, it was known that DA 6034 has local gastroprotective effect as well as anti-inflammatory effect for inflammatory bowel disease. It was thought that its gastroprotective effect would be limited by its low solubility in gastric region because of its low solubility in acidic condition, less than 1 g/ml, without any drug delivery technology such as solubilization or controlled release. EFMS consists of matrix forming additives such as gas generating agent, swelling polymer, erosion polymer, solubilizer, and it could be pressed to a tablet by mixing lubricants. It was designed to have floating property when it was hydrated with acidic gastric fluid, and solubilized drug in hydrated gel matrix could be released continuously with enhanced solubility by erosion of gel matrix. Amorphous solid dispersion of DA 6034 with hydrophilic carrier such as HPMC, PVP, HP-beta CD, SLS was produced by spray drying or using high speed mixer for the purpose of increasing solubility at acidic condition, and it was characterized by XRD and DSC. Solubility of spray dried DA 6034 was modestly enhanced in buffer solutions, from pH 1.2 to pH 4, and precipitation was generated by recrystallization with time for solid dispersions with HP-beta CD, SLS. Solubility of solid dispersed DA 6034 was highly enhanced than untreated DA 6034 in buffer solution of pH 4. EFMS tablet containing DA 6034 or its solid dispersion was produced by wet granulation method. Dissolution rate of EFMS tablets in acidic buffer solutions was highly enhanced than that of control tablet, which was prepared by conventional method without any other solubilization or controlled release technology. There was only slight difference in dissolution rate between EFMS tablets containing DA 6034 or its solid dispersion, and it showed that enhanced dissolution was acquired not only by formation of solid dispersion but also by solubilizers in gel matrix. The formulation of EFMS tablet of DA 6034 was optimized by adjusting the species and amount of gas generating agent, solubilizers and erosion polymers, and stability and gastroprotective effect with ulcer induced beagle was confirmed by evaluated for optimized formulation of EFMS tablet of DA 6034. In conclusion, controlled release profile with increased solubility was acquired in acidic buffer solution with EFMS tablet of DA 6034, and clinical study with human will be carried out with this EFMS of DA 6034.

DA-6034의 polymorphic transformation에 關한 硏究 : Study on polymorphic transformation of DA-6034

서현옥 德成女子大學校 大學院 2006 국내박사

DA-6034 는 eupatilin의 합성유도체로서 그 기전이 분명히 밝혀지지는 않았으나 실험적으로 염증성 대장 증후군에 유효성을 갖는 신약후보물질이다. 신약개발과정 중의 한 단계로 DA-6034 에 대한 polymorphism 연구가 수행되어, 아홉 개의 pseudopolymorph 와 한개의 polymorph 가 분리된 바 있다. 본 연구에서는 선행된 연구에 이어 각 modification의 재현성 및 transformation여부를 확인하고자 하였다. 선행연구에서 제시된 10 가지 modification 중 6 가지 pseudo-polymorph 에 대하여 상대습도와 온도에 대하여 일정한 조건에 보관하면서 시간에 따른 형 전환과정을 살펴보았다. 내부 구조의 변화를 알아보기 위해 XRPD pattern 을 일정한 시간간격으로 분석하였고, solvate 내부의 변화를 측정하기 위해 DSC 와 TG 로 열분석을 시행하였다. 결과적으로 본 실험에 이용된 대부분의 modification 들은 온도보다는 습도에 민감하게 반응하여 상대습도가 높은 환경에서는 원물질인 monohydrate 로 전환되었다. 0% RH 조건에서는 결정성이 감소하여 amorphous solid 를 형성하였으며, 고온 조건 하에서도 같은 결과를 나타내었다. 예외적으로 Form 11 은 비교적 안정하여 생성 당시의 결정구조를 유지하였다. 주목할만한 점은 Form 8 의 형 전환으로, 0% RH 조건에서는 다른 modification 들과 마찬가지로 amorphous solid 를 형성하였으나 고습 조건하에서 선행연구에서 발견되지 못한 미지의 형으로 전환되었으며 이에 대한 연구가 계속 진행되어야 하겠다. DA-6034, 7-carboxymethyloxy-3', 4', 5-trimethoxyflavone, is a new drug candidate known to possess anti-inflammatory activity. The study on polymorphism of DA-6034 was already accomplished. However, it is needed to examine further those polymorphs especially on stability and transformation. In this study, it was investigated that the stability of six modifications depending on temperature and relative humidity. Six solvates, with DMSO, 2-butanone, acetic acid, n-butyl alcohol, and benzyl alcohol, were isolated by the method of previous study and were kept in different conditions : in 0%, 52%, and 95% RH at room temperature to investigate the effect of relative humidity and at RT, 40℃, and 60℃ to investigate that of temperature. XRPD, DSC, and TG analysis were used to examine the state of modifications. Consequently, solvates with DMSO, 2-butanone, and n-butyl alcohol are transformed to the original form of DA-6034 only in a week by the effect of relative humidity rather than high temperature, and all solvates except the solvate with benzyl alcohol became amorphous solids under the condition of 0% RH. A solvate with benzyl alcohol is stable enough to keep its form even under high-humidity condition. Most interesting result is the transformation of the solvate with acetic acid, which is transformed to an unknown modification.

DA-6034가 소장염증 모델에서 장투과도에 미치는 영향

곽동신 한양대학교 2014 국내석사

Background & Aim: DA-6034 is known to have antiinflammatory activities and exhibits cytoprotective effects in acute gastric injury model. Here we would like to investigate the possible protective role of DA-6034 in indomethacin induced small intestinal damage model and their effect on intestinal permeability. Methods: Small intestinal damage was induced with indomethacin (15mg/kg). Rats in the treatment group received DA-6034 (30 mg/kg) from day 0 to 2, and indomethacin from day 1 to 2 through intragastric lavage. On the contrary, rats in the control group received only indomethacin from day 1 to 2. On the fourth day, Rats were sacrificed, and small intestine was examined to compare the severity of inflammation in both groups by H&E stain. Intestinal permeability was evaluated by Fluorescein isothiocyanate (FITC)-labeled dextran. Western blot was performed to comfirm the association of DA-6034 with ERK pathway in the prevention of small intestinal injury. Results: Intestinal permeability was lower in treatment group compared to control group (16204.2±13832.1 vs 30403.7±24012.6, p value <0.05). The inflammation score in the treatment group was lower compared to that in the control group but statistically insignificant (2.6±0.6 vs 2.9±0.3, p=0.11). When the extent of hemorrhage was compared, hemorrhagic lesion in treatment group was broader than that of control group, but was not statistically significant (45±45.5 vs. 43.0±33.8, P=0.88) DA-6034 enhanced the phosphorylation of extracellular signal-regulated kinase (p-ERK) expression in small bowel mucosa (107.4±40.2 vs 64.2±20.4 P value <0.05) and intestinal permeability was negatively correlated with density of ERK expression (r=-0.309, p=0.096). Conclusion: DA-6034 decrease intestinal permeability in indomethacin induced intestinal injury model via P-ERK pathway.

인간 대장 점막 세포주에서 DA-6034가 lipopolysaccharide 및 tumor necrosis facor-α [실제는 factor-α] 자극에 의한 NF-kB의 활성화에 미치는 영향

김지원 서울대학교 대학원 2004 국내박사

상피세포에서 DA-6034에 의해 유도된 세포 내 칼슘농도 증가

지혜원 연세대학교 대학원 2014 국내석사

Pharmacokinetics and tolerability of eupatilin-derived compound, topical ocular GLH8NDE

정지현 서울대학교 대학원 2022 국내석사

Introduction: Dry eye disease (DED) is a multi-factor disease on the surface of the eye that is accompanied by inflammation of the eye surface, causing eye discomfort, visual impairment, and instability of the tear film. Along with the daily use of smartphones and frequent occurrence of fine dust, the prevalence rate of the disease is increasing as related to the quality of life. Cyclosporin, steroids, and tetracycline have been currently used to treat DED, but they are not fundamental cause-and-effect treatments based on the etiology of DED. Accordingly, patients' satisfaction with the present therapies has been low in addition to the safety issues reported by a high frequency of side effects. Therefore, it is necessary to develop a treatment that is safe and could more fundamentally treat DED. Based on the need to develop a fundamental DED treatment, GLH8NDE is considered as a promising treatment with its' strong mucosal protection and anti-inflammatory pharmacological actions. GLH8NDE is 7-carboxymethyloxy-3',4',5'-trimethoxy flavone, a derivative of eupatilin, newly synthesized from the flavonoid family Artemisia asiatica, and an oral product has been currently used as a treatment for gastritis and gastric ulcers. The tolerability, and pharmacokinetic properties of the oral drug have been evaluated in healthy adults, but those of the topical ocular drug have not been assessed. Therefore, this study explored the pharmacokinetic properties of GLH8NDE when administered to healthy adults, and evaluated tolerability based on the systemic exposure and the occurrence of systemic adverse events with the fact that GLH8NDE is mainly distributed to the cornea and conjunctiva, showing excellent effects in various experimentally-induced animal DED models through increasing mucin secretion and anti-inflammatory effects. In addition, the difference in the systemic exposure of GLH8NDE between Koreans and Caucasians was also investigated through comparison of the pharmacokinetic parameters. Methods: A randomized, double-masked, placebo-controlled clinical trial was conducted in 30 healthy Korean and 10 healthy Caucasian males to explore the pharmacokinetic properties and evaluate tolerability of GLH8NDE. Ten subjects in each cohort were randomly administered placebo or GLH8NDE 5 % (w/v) to both eyes at the scheduled dosing time. This study consisted of 2 parts, single- and multiple-day administrations, based on the daily doses. After 62-hour washout period followed by single-day administration the subjects proceeded to multiple-day administrations of the same dose for 7 consecutive days. The subjects in cohort A were administered 9 mg per day at intervals of 5 hours, and the subjects in B, C, and D were given 18, 36, and 36 mg per day, respectively, at intervals of 2 hours. The subjects in cohort A and B were administered one drop each to both eyes, and who in C and D were given two drops each to both eyes. All of them were Koreans except for who were in cohort D. The subjects in cohort C and D received the same dose, but cohort C consisted of Koreans and cohort D included Caucasians. After administration the predetermined physical examination was conducted including the ophthalmic symptom evaluation and the ophthalmic test. The safety evaluation was conducted under the consultation of an ophthalmologist after reviewing the incidence of adverse events, physical examination, vital signs, 12-lead ECG, and the clinical laboratory test. Blood collection was conducted at predetermined time points until 24 hours after single-day and the last administration of multiple-day administrations to evaluate the pharmacokinetic properties of GLH8NDE. A total of 40 demographic information, vital signs, clinical laboratory testing, serum concentration of GLH8NDE, ophthalmic symptom evaluation, ophthalmic testing, and adverse events coded MedDRA Ver. 23.0 were collected for analysis. Results: We found that the systemic exposure of GLH8NDE after administration in healthy adults was negligible. The systemic exposure increased proportionally as administration dose was escalated. We also found that pharmacokinetic parameters showed dose-dependent changes. Furthermore, the systemic exposure to GLH8NDE was dose proportional without drug accumulation. In this study, a total of 32 adverse events occurred in 17 out of 40 subjects who received GLH8NDE or placebo at least once. All adverse events were mild in intensity. A total of 28 cases of them were determined to be in relation to the study drugs, and 22 were ocular-related adverse events. A total of 6 systemic adverse events were reported in 5 subjects who received GLH8NDE. All systemic adverse events were temporarily mild and recovered without further medical treatments. In addition, the incidence of adverse events did not increase with dose escalation, but rather they occurred more frequently in lower dose cohorts. No abnormal findings or changes regarding clinical significance were observed in ophthalmic tests, clinical laboratory tests, vital signs, physical examination, and 12-lead ECG tests. The systemic exposure to GLH8NDE showed racial differences. In the same dose groups, systemic exposure to GLH8NDE was higher in Koreans than in Caucasians. AUClast and Cmax in Caucasians decreased by 2.09 and 1.33 times respectively when compared to those in Koreans. Conclusions: This study was the first to explore the pharmacokinetic properties of GLH8NDE and evaluate its' tolerability in health adults. The systemic exposure to GLH8NDE was negligible and no persistent and severe systemic adverse events occurred when administered to healthy adults. In addition, it was confirmed that the systemic exposure to GLH8NDE was higher in Koreans than in Caucasians. The above results suggest that GLH8NDE is reliably safe and tolerated when topically administered as an ophthalmic agent. Additionally, this study provides the information about racial differences in the systemic exposure to GLH8NDE when topically administered, which should be considered in further studies. 서론: 안구 건조증은 안구 표면의 염증이 동반되어 눈의 불쾌감, 시각장애, 눈물막의 불안정성을 유발하는 눈물과 안구 표면의 다요소적 질환이다. 안구건조증은 일반 성인의 약 17%~25%가 가지고 있을 정도로 매우 흔하며, 연령이 증가함에 따라 비례하여 증가한다. 스마트폰 사용의 일상화와 더불어 미세먼지 등의 공기 질이 나빠지면서 삶의 질과 관련된 질환으로 안구건조증의 발병율은 점점 증가하는 추세이다. 사이클로스포린, 스테로이드, 테트라사이클린이 안구 건조증의 치료에 사용이 되고 있으나 안구건조증이라는 병인론에 근거하였을 때 근본적인 원인 치료제가 아닌 단순히 증상만을 완화시키는 대증요법제 이므로 환자 만족도가 낮고, 부작용 발현 빈도가 많이 보고 되어 안전성에 우려가 있다. 따라서 안전하면서도 보다 근본적으로 안구건조증을 치료할 수 있는 치료제 개발이 요구되고 있다. 이러한 근본적인 안구건조증 치료제 개발의 필요성에 근거하면, 강력한 점막보호작용 및 항염증 약리적 작용을 갖는 GLH8NDE가 안구건조증의 유망한 치료제로써 개발가치가 충분할 것으로 사료된다. GLH8NDE는 쑥 추출물(Artemisia asiatica)에서 신규 합성한 flavonoid 계열의 eupatilin 유도체로써 같은 성분의 경구용 제제가 위염 및 위궤양 치료제로 현재 쓰이고 있다. 건강한 성인을 대상으로 경구용 제제의 안전성, 내약성 및 약동학적 특성을 평가하여 우수한 내약성 및 안전성을 확인한 바 있으나 점안제의 안전성, 내약성 및 약동학적 특성을 평가한 바는 없다. 따라서 본 연구에서는 GLH8NDE 점안제가 실험적으로 유발한 다양한 안구건조증에서 효과와 안정성이 확보된 물질로써 점적 후 안구조직인 각막과 결막에 주로 분포하며 점액분비 증가작용, 항염증작용, 상처치유작용 및 수분분비 증가작용을 통하여 우수한 효과를 나타냈다는 점에 착안하여 GLH8NDE 점안제를 건강한 성인에게 투여하여 약물의 약동학적 특성을 살펴보고, 체내 노출 정도와 전신 이상 반응 출현의 빈도를 근거로 내약성을 평가하고자 한다. 또한, 산출된 약동학적 파라미터 비교를 통하여 같은 용량군에서 한국인과 서양인의 체내 GLH8NDE 노출 차이를 확인하고자 하였다. 방법: 본 연구는 GLH8NDE 점안제의 약동학적 특성, 안전성 및 내약성을 평가하기 위하여 무작위배정, 이중눈가림, 위약대조 임상시험을 30명의 건강한 한국인 남성과 10명의 서양인 남성에서 수행하였다. 용량군 당 10명씩 무작위 배정하여 배정된 용량군에 따라 예정된 투약 시간에 GLH8NDE 점안액 5 %(w/v) 또는 위약을 양안에 점안하였다. 1일 복용량을 기준으로 단회 및 반복투여로 진행하여, 단회 투여 후 62 시간의 휴약기를 거쳐 같은 용량으로 7일간 반복투여를 진행하였다. 용량군 A는 5 시간 간격으로 하루에 9 mg를 투여 받았고, 용량군 B, C, D는 2 시간 간격으로 하루에 각각 18 mg, 36 mg, 36 mg를 투여 받았다. 용량군 A와 B는 양안에 각각 1방울, 용량군 C와 D는 양안에 각각 2방울씩 점안하였다. 용량군 D를 제외한 나머지 용량군은 모두 한국인이었다. 용량군 C와 D는 동일한 용량을 투여 받았으나 용량군 C는 한국인, 용량군 D는 서양인이었다. 점적 후 안과적 증상 평가 및 안과적 검사를 포함한 정해진 검사를 시행하였고, 안전성 평가는 이상반응의 발생 양상을 비롯하여 신체검사, 활력징후, 심전도검사, 임상실험실검사, 안과적 검사 결과 등을 총괄적으로 검토하여 안과 전문의의 자문 하에 이루어졌다. 또한, GLH8NDE의 약동학적 특성을 평가하기 위해 단회 투여 후와 마지막 반복투여 후 24 시간까지 정해진 시간에 채혈을 시행하였다. 혈중 GLH8NDE의 농도 분석 결과로 약동학적 파라미터를 산출하여 투여 후 체내 GLH8NDE 노출 정도를 살펴보고 이와 더불어 수집된 이상반응 출현의 빈도를 통하여 GLH8NDE 점안제의 안전성과 내약성을 평가하기 위하여 투여를 진행한 총 40명의 인구학적 정보, 활력징후 결과, 임상실험실 검사 결과, 혈중 GLH8NDE의 농도, GLH8NDE 투여 후 안과적 증상 평가 결과, 안과적 검사 결과, MedDRA Ver. 23.0으로 코딩한 이상반응의 자료를 수집하였다. 또한, 같은 용량군에서 한국인과 서양인의 체내 GLH8NDE 노출에 차이가 있는지 알아보기 위하여 두 그룹에서 산출된 약동학적 파라미터를 비교 분석하였다. 결과: 건강한 성인에서 GLH8NDE 점안제 투여 후 체내 GLH8NDE 노출은 무시할 만할 정도로 적음을 확인하였다. 체내 노출은 용량이 증가함에 따라 용량비례적으로 증가하였다. 체내 노출뿐만 아니라 약동학적 파라미터도 용량 의존적인 변화를 보이는 것을 확인하였다. 또한, 체내 GLH8NDE 노출은 반복투여에 따른 축적을 보이지 않았다. 본 연구에서 GLH8NDE 점안제 또는 플라시보를 1회 이상 투여 받은 총 40명의 대상자 중 17 명에서 총 32 건의 이상반응이 발생하였다. 발견된 모든 이상반응은 중대하지 않은 이상반응인 것을 확인하였다. 그 중 총 28 건이 임상시험용의약품과 관련하여 인과관계가 있다고 판단하였고, 22 건이 안구관련 이상반응이었다. 전신 이상반응은 GLH8NDE 점안제를 투여 받은 5명에서 6건이 보고되었다. 발생한 전신 이상반응은 일시적인 경증 반응이었고 의학적 처치 없이 회복되었다. 또한, 발견된 이상반응은 용량 증가에 따라 발생빈도가 증가하지 않았으며 오히려 저용량 군에서 더 빈번하게 발생하였음을 확인하였다. 안과적 검사, 임상실험실검사, 활력징후, 신체검사 및 심전도검사에서 임상적 의미를 갖는 이상소견이나 변화는 관찰되지 않았다. 그리고 체내 GLH8NDE 노출은 인종간 차이를 보였다. 같은 용량군에서 체내 GLH8NDE 노출은 AUClast 기준 2.09 배, Cmax 기준 1.33 배 서양인에서 보다 한국인에서 더 높게 나타났다. 결론: 본 연구는 건강한 성인을 대상으로 GLH8NDE 점안제의 약동학적 특성을 살펴보고 안전성과 내약성을 평가한 첫번째 연구이다. 점적 시 체내 GLH8NDE 노출은 무시할 만할 정도로 적었으며 지속적이고 중증의 전신이상반응은 발생하지 않았다. 또한, 체내 GLH8NDE 노출은 서양인 보다 한국인에서 더 높게 관찰됨을 확인할 수 있었다. 이상의 결과는 GLH8NDE를 점안제로 투여 시 안전성과 내약성이 양호하다는 근거를 제시하며, 점적 시 서양인과 한국인에서 체내 GLH8NDE 노출이 다르므로 추가 연구 시 인종간 차이를 고려해야 함을 시사한다.