초록 ▼

1) 연구목표: 기존 골다공증치료제의 clinical unmet needs를 극복할 수 있는 신개념 Dual 기전(조골세포 촉진 및 파골세포 억제)을 가지는 골다공증 치료제 후보 물질 SP-35454의 개발타당성을 평가하고 비임상독성, 약효평가, 약동력학, 제형연구, 후속후보물질 선정 등을 진행 후 임상1상 신청을 목표로 함.
2) 연구내용: TAZ를 통해 골형성 촉진과 골흡수 억제의 Dual 기전을 규명하고 in vitro/in vivo 약효평가연구, 1년 장기 효력시험, 비임상 반복투여 독성시험 및 임상시험 원료 합성법 개

1) 연구목표: 기존 골다공증치료제의 clinical unmet needs를 극복할 수 있는 신개념 Dual 기전(조골세포 촉진 및 파골세포 억제)을 가지는 골다공증 치료제 후보 물질 SP-35454의 개발타당성을 평가하고 비임상독성, 약효평가, 약동력학, 제형연구, 후속후보물질 선정 등을 진행 후 임상1상 신청을 목표로 함.
2) 연구내용: TAZ를 통해 골형성 촉진과 골흡수 억제의 Dual 기전을 규명하고 in vitro/in vivo 약효평가연구, 1년 장기 효력시험, 비임상 반복투여 독성시험 및 임상시험 원료 합성법 개발 및 생산 등을 진행하여 유럽에서 임상1상 신청 준비를 완료하였음. 추가적으로 후속후보물질을 선정함.
3) 연구결과: 새로운 개념의 골다공증 치료제인 SP-35454는 골다공증 질환 동물 모델에서 기존 약물대비 우수한 개선 효과를 보였음. 특히, 골흡수 억제뿐만 아니라 TAZ를 통한 골형성 촉진을 통해 bone quality를 높이는 차별화된 결과를 보였음. 비임상 약리독성 시험 및 Rat, Dog 4주 반복 투여 독성시험을 통해 고농도에서의 안전성을 확보하였으며 마일스톤 1의 비임상 시험 결과를 토대로 유럽(네덜란드)에서의 임상 1상 준비를 마쳤음. 추가적으로 우수후속후보물질 6종을 선정함. SP-35454는 기존 골다공증치료 약물의 clinical unmet needs를 극복할 수 있는 새로운 기전의 치료제로서 개발 가능성이 높은 후보물질이라 판단됨.

(출처 : 보고서 요약서 3p)

Abstract ▼

IV. Result
1. Solid state analysis
Crystalline salts was analyzed in screening experiments. The solid-state properties (XRD, PLM, DSC, TGA, DVS) and solubility were investigated. The solubility of hydrochloride and besylate form in water were increased comparing with freebase form. But the sta

IV. Result
1. Solid state analysis
Crystalline salts was analyzed in screening experiments. The solid-state properties (XRD, PLM, DSC, TGA, DVS) and solubility were investigated. The solubility of hydrochloride and besylate form in water were increased comparing with freebase form. But the stability was decreased. So. the free base form was chosen.

2. Non-clinical API synthesis (10kg scale-up)
11.6 kg SP-35454 API was synthesized for non clinical study through optimized synthesis process (7 step).

3. Synthetic process development and improvement
Synthetic process was developed and improved to solve some problems occurred in synthesis process.

4. Metabolites study
Because the metabolite of SP-35454 was identified in human study, the metabolite study was performed. Metabolite M8-1 was synthesized and the structure was analyzed through various spectrascopy.

5. Validation protocol development and stability study
SP-35454 was micronized to improve solubility for bioavailability and assessed in vivo study. Micronized and non-micronized SP-35454 were administered to dog. And dog plasma concentration was analyzed in Pharmacokinetic study. Micronized SP-35454 in Cmax, AUC parameter was 3 times higher than non-micronized SP-35454. The interaction between ingredient and excipient was analyzed to assess the stability of ingredient. Base on results, a suitable excipient was chosen.

6. Clinical API synthesis (10kg scale-up)
Synthetic process for Clinical API was set-up through improvement (8 steps). 10 kg of clinical API for clinical phase 1 trial was synthesized.

7. Drug metabolism study (in vivo )
Metabolite was detected more than 20% in human study. The metabolite will be managed in further study. And the study of metabolite will be performed consistently.

8. Formulation analysis in aqueous solution/suspensions
The accuracy and homogeneity of SP-35454 were determined in 0.5% Methyl Cellulose (1500 cps viscosity). And Oral dosing formulation was chosen.

9. LC-MS/MS bioanalytical support Rat & Dog
SP-35454 was investigated in rat and dog plasma to develop the bioanalytical process. After protein precipitation extraction, the samples are analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The range of the assay was 10.0-10000 ng/mL (SP-35454).

10. Rat tolerability study
Administration of SP-35454 by once daily oral gavage for 3 days at 500 mg/kg/day followed by 7 days at 1000 mg/kg/day was considered to be well tolerated but was associated with reduced body weight gain and body weight loss, and low food intake. Based on these findings, a dose level of 1000 mg/kg/day was determined as a high dose level for 4 week repeat toxicity study.

11. 4 week rat toxicity + 2 week recovery
Administration of SP-35454 once daily by oral gavage was well tolerated in rats at dose levels up to 1000 mg/kg/day. Based on these results, NOAEL was determined to be 1000 mg/kg/day.

12. Dog tolerability study
Dosing of SP-35454 for up to 7 days at escalating dose levels of 300, 600 and 1000 mg/kg/day was not associated with any effects on body weight, food consumption, electrocardiology, clinical pathology, gross pathology or organ weights. Based on the results of this study, SP-35454 was considered to be well tolerated in the male and female beagle dog at dose levels up to 1000 mg/kg/day for up to 7 days, and that the dose levels used on this study was determined for 4 week repeat toxicity study.

13. 4 week dog toxicity + 2 week recovery
Administration of SP-35454 once daily by oral gavage was well tolerated in dogs at dose levels up to 1000 mg/kg/day and the observed findings were considered not to be adverse. Based on results, NOAEL was determined to be 1000 mg/kg/day.

14. Safety Pharmacology
1) Central nervous system
Based on results, the no-observed-effect level (NOEL) was considered to be 100 mg/kg. However, due to the reversibility of the observed findings the NOAEL was determined to be 1000 mg/kg.
2) Respiratory system
No relevant effects on respiratory parameters were observed after administration of SP-35454 at any dose levels tested. Based on results, the no-observed-effect-level (NOEL) was determined to be 1000 mg/kg.
3) Cardiovascular system
① In vitro hERG (human ether related-a-go-go)
SP-35454 was applied at achieved mean concentrations of 0.07, 0.09, 0.30 and 1.18 μM (nominal concentrations of 0.1, 0.3, 1 and 3 μM, respectively). The significant inhibition of the hERG tail current was observed at 0.30 and 1.18 μM (1.18 μM equals 0.63 μg/ml). The inhibition rate of 45.9% was observed. But IC50 value could not be identified.
② In vivo (A cardiovascular (CV) study)
SP-35454 induced no adverse or relevant changes in cardiac hemodynamics or electrophysiology in dogs following single oral administration of SP-35454 at the dose-levels of 100, 300 and 1000 mg/kg. Based on results, the no observed effect level (NOEL) was determined to be 1000 mg/kg.

15. Clinical trial protocol
Clinical trial protocol was written for clinical phase 1 trial. The trial will be a double-blind, randomized, placebo-controlled, single-center trial. The cross over design will be applied to Single dose part and the parallel group design will be applied to Multi dose part. The safety, tolerability and pharmacokinetics will be evaluated in clinical study. Eligible subjects should fulfill all the inclusion criteria and none of the exclusion criteria.

16. IND application and submission
IND file was prepared and submitted for CTA approval. CTA will be approved in March 2015. And clinical trial of SP-35454 will be started in April 2015.

17. Mode of action establishment
Mechanism study was performed. Pharmacological data indicate that SP-35454 has a dual action: stimulation of bone formation by modulating the ‘Transcriptional co-activator with PDZ-binding motif’ (TAZ) and inhibition of bone resorption. Base on results, SP-35454 does not effect on the expression of TAZ, but activates the translocation of TAZ to nucleus.

18. Relation study of TAZ and carcinogenesis
The effects of TAZ modulation on carcinogenesis was investigated. SP-35454 did not induce any carcinogenesis in in vitro study using MCF7 and MDA-MB-231 cell line. And the effect on carcinogenesis will be evaluated through long term toxicity study in milestone 2.

19. Biomarker development
New biomarker was investigated to identify the efficacy. SP-8203 related-biomarker (P1NP, CTX, ALP and leptin) were tested.

20. Efficacy study in OVX rat model
The administration of SP-35454 at concentration of 1, 5, 10 mg/kg showed the dual effects of born resorption inhibition and born formation activation in 1 year long term efficacy study using OVX rat model. SP-35454 also showed the dual effects in short term study using OVX rat model.

21. Back-up compound synthesis and evaluation
About 300 back-up compounds were synthesized based on structure. We evaluated the effect of back-up compounds in in vivo studies included zebrafish model. Based on results, we selected twenty back-up compounds more effective than SP-35454. Finally, We chose 6 candidate compounds for follow-up study..

(출처 : Summary 10p)

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 4
• SUMMARY ... 9
• CONTENTS ... 14
• 목차 ... 16
• 제1장 연구개발과제의 개요 ... 18
• 제1절 의료계 수요 ... 18
• 1. 경제적, 산업적 중요성 및 필요성 ... 18
• 제2절 타겟 선정 배경 ... 22
• 제2장 국내외 기술개발 현황 ... 24
• 제1절 기존의 골다공증 치료 약물 ... 24
• 제2절 연구의 향후 전망 ... 25
• 1. 후보물질의 독창성 ... 25
• 2. 후보물질의 경쟁력 ... 26
• 제3장 연구개발수행 내용 및 결과 ... 29
• 제1절 CMC 연구 ... 29
• 1.Solid state 연구 (salt selection) ... 29
• 2.비임상용 원료합성 (10 kg scale-up) ... 33
• 3.합성 process 개발 및 합성법 최적화 연구 ... 35
• 4.대사체 합성 연구 ... 46
• 5.비임상용 원료 분석법 개발 및 안정성 연구 ... 48
• 6.임상용 원료 합성 (10 kg scale-up) ... 70
• 제2절 비임상연구 ... 72
• 1.종간 대사체 확인시험 (in vitro) ... 72
• 2.Formulation analysis in aqueous solution/suspensions ... 84
• 3.LC-MS/MS bioanalytical support ... 86
• 4.Rat tolerability study ... 92
• 5. 4 week rat toxicity + 2 week recovery ... 103
• 6.Dog Tolerability Study ... 120
• 7. 4 Week dog toxicity + 2 week recovery ... 130
• 8.안전성 약리 시험 ... 147
• 제3절 IND 신청 ... 167
• 1.임상시험 프로토콜 작성 ... 167
• 2.IND file 작성 및 신청 ... 173
• 제4절 약효 및 기전 연구 ... 177
• 1.Mode of action 확립 ... 177
• 2.TAZ와 암유발과의 관계 연구 ... 188
• 3.효능검증 바이오 마커 개발 ... 193
• 4.동물모델을 이용한 약효 평가 (노령쥐에서의 효력시험) ... 196
• 5. 동물모델을 이용한 약효평가 (OVX 모델에서의 효력시험, 기타동물모델에서의 효력시험) ... 224
• 제5절 Back-up 화합물 합성 및 평가 ... 232
• 1.Back-up 화합물 합성 및 평가 (한국화학연구원) ... 232
• 2.Back-up 화합물 합성 및 평가 (신풍제약) ... 249
• 제4장 목표달성도 및 관련분야에의 기여도 ... 264
• 제1절 목표달성도 ... 264
• 제2절 특허 확보 ... 270
• 제5장 연구개발결과의 활용계획 ... 271
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 272
• 제7장 연구시설 · 장비 현황 ... 273
• 제8장 참고문헌 ... 274
• 끝페이지 ... 276