초록 ▼

Ⅳ. 연구개발결과
가. 비임상 독성 시험
비임상 독성시험
일반독성 시험
▶ 랫드 단회투여 독성평가
개략의 치사량(ALD), 경구: 암컷 1000 mg/kg, 수컷 2000 mg/kg. 개략의 치사량 (ALD), 정맥: 암컷 및 수컷 200 mg/kg 이상
▶ 랫드 2주 반복투여 독성평가,　최대내성용량 (MTD): 500 mg/kg/day
▶ 랫드 4주 반복투여 독성평가, 최대무해용량 (NOAEL): 250 mg/kg/day
▶ 원숭이 단회투여 독성평가 (단회 용량증가)
반복투여시험 용

Ⅳ. 연구개발결과
가. 비임상 독성 시험
비임상 독성시험
일반독성 시험
▶ 랫드 단회투여 독성평가
개략의 치사량(ALD), 경구: 암컷 1000 mg/kg, 수컷 2000 mg/kg. 개략의 치사량 (ALD), 정맥: 암컷 및 수컷 200 mg/kg 이상
▶ 랫드 2주 반복투여 독성평가,　최대내성용량 (MTD): 500 mg/kg/day
▶ 랫드 4주 반복투여 독성평가, 최대무해용량 (NOAEL): 250 mg/kg/day
▶ 원숭이 단회투여 독성평가 (단회 용량증가)
반복투여시험 용량설정
▶ 원숭이 2주 반복투여 독성평가
최대내성용량 (MTD): 100 mg/kg/day
▶ 원숭이 4주 반복투여 독성평가
최대무해용량 (NOAEL): 80 mg/kg/day

유전독성 시험
▶ 복귀돌연변이 시험 (in vitro, AMES) : 음성
▶ 소핵분석 (in vivo, MNT)：음성
▶ Comet assay (in vivo)：음성

안전성 약리 시험
▶ 심혈관계
hERG : IC50 > 100 uM
Telemetry (혈압, 심박수/심전도): 80 mg/kg까지 영향 없음
▶ 호흡기계
호흡률/호흡기능: 500 mg/kg까지 호흡기계 영향 없음
▶ 중추신경계
FOB: 500 mg/kg까지 중추신경계 영향 없음

조제물 분석법 셋업 및 평가
▶ 조제물 분석법 셋업 및 평가
셋업 완료
4주 랫드 및 원숭이 시험의 조제물 분석 완료
▶ 생체시료 셋업 및 평가
셋업 완료
4주 랫드 및 원숭이 시험의 생체시료 분석 완료

약물동태 시험
In vitro ADME 평가
▶ Caco-2 permeability 시험 :높은 투과도 확인, p-gp 기질 가능성
▶ 혈장 내 안정성 시험：5종에서 반감기 277분 상회
▶ 마이크로좀 안정성 시험：사람 간 마이크로좀에서 30~40% 대사됨
▶ 간세포 안정성 시험：5종의 반감기가 457분 상회, 혈장에서 안정
▶ 대사체 확인 시험 : 원숭이 및 비글에서 M1 대사체 확인
▶ UGT phenotyping 시험
7종 효소 존재 하 배양 시 잔여량 96% 이상　UGT에 의해 대사되지 않음
▶ 혈장 단백 결합 시험
중등도 이하의 결합도 확인, 유의한 농도 의존성 없음
▶ Blood to plasma partition assay
모든 종에서 적혈구와 혈장에 동일하게 분포
▶ CYP450 저해 시험
CYP1A2, CYP2B6, CYP2D6, CYP2C8, CYP2C9, CYP2C19의 최대 저해 비율은 9% 미만
CYP3A4의 최대 저해 비율은 19.72% (CYP3A4의 약한 저해제)
▶ CYP450 유도 시험
CYP1A2, CYP2B6, CYP3A4의 효소 유도 없음 (최대 10μM)
▶ GSH trapping study : GSH 결합체 발견되지 않음
▶ UGT inhibition enzyme test
50μM에서 UGT1A9(38.8%), UGT2B7(39.1%)의 활성을 저해 0~50μM에서 UGT1A1, UGT1A3 저해 없음, UGT1A4, UGT1A6에 약간의 저해 효과

약물동태 시험
In vivo 약물동태
▶ 랫드 약물동태 평가
Cmax 6671±1715 ng/mL, Tmax 1~2 hr, AUC(0-∞) 33328±2892 hr*ng/mL, BA 70.0 ± 6.1%
용량-비례, 축적 없음, 성별차이 없음, 식이 효과 없음
▶ 원숭이 약물동태 평가
Cmax 2712±401 ng/mL, Tmax 0.5~2 hr, AUC(0-∞) 12554±346 hr*ng/mL, BA 84.4 ± 2.6% 용량-비례, 축적 없음, 성별차이 없음, 식이 효과 없음

방사선 표지화합물
▶ 14C-JW1345 합성 완료, 적합성 확인

QWBA
▶ Autoradiography/Mass balance
다른 장기에 비하여 위장관에 높게 분포. 체내에서 빠르게 소실. 시험한 조직 중 눈, 뇌, 백색지방조직에 가장 적게 분포.

효능/추가 적응증
건선
▶ Imiquimod-induced psoriasis-like model 1차 평가
BID. Epidermal thickness를 유의적으로 억제 (양성대조군과 유사 효능)　등 두께를 유의적으로 억제. 각질 발현량 유의적으로 억제
▶ Imiquimod-induced psoriasis-like model 2차 평가
BID 및 QD 조건에서 Epidermal thickness 및 등 두께 유의적으로 억제 (양성대조군과 유사 효능)/ 각질 발현량을 유의적으로 억제
▶ Human skin transplant psoriasis model
BID 조건에서 Epidermal ridge thickness와 Ki67 index를 유의적으로 억제 (양성대조군과 유사 효능)

나. 합성
- JW1345(API)의 합성공정 최적화 연구를 진행하였으며 각 합성단계 별 DoE연구를 통한 QbD 적용으로 반응 최적화 연구를 진행함.
- 합성단계 별 생성가능성이 있는 유연물질을 합성, 정제하여 구조를 규명.
- 유연물질의 fate-map을 작성하고 유연물질의 기준을 설정.　
- 합성공정 최적화 연구를 통해 얻은 공정으로 총 6회의 scale-up을 진행.
- Scale-up으로 얻은 API는 제제연구와 비임상독성 시험용으로 사용되었으며 임상시험용 API는 cGMP site인 에스티팜에서 JW중외제약이 개발한 공정으로 진행함.
- 총 11.69 kg의 JW1345(API)를 얻었으며 모든 분석 항목에서 적합임을 확인 후 제조된 API는 임상약 제조가 진행될 WuXi AppTec으로 송부함.

다. 제제연구 및 DP 생산
(1) DP 분석법 set-up 및 안정성 분석 (JW1601정 시험방법)
(2) 연구용배치 안정성시험 계획 요약표
(3) 안정성시험 결과
장기안정성시험은 전 용량 6 개월까지 평가가 완료되었으며 현재까지의 결과 유의적인 변화는 없었고 최종 36개월까지 진행할 예정임. 가속안정성시험은 전 용량 최종 6개월까지 평가가 완료되었으며 모든 결과가 기준에 적합하였음.

라. 완제의약품 개발
- JW1601 정은 경구투여가 가능한 아토피 치료목적의 임상시험약으로 개발하기 위해 범용적으로 사용되는 정제 제형으로 개발을 검토하였음.
- JW1601 정의 개발 전략은 임상 용량별 안전성과 유효성을 확인하기 위한 속방형 제제로서 용량별 용출양상이 동등하고 안정한 제제로 개발하는 것이었으며 임상 1상 시험을 위해 FR1345로서 세 가지 용량을 선정하여 전 용량 동일한 질량을 가진 정제로서 유사한 성상을 나타내도록 밝은 분홍색의 원형 필름코팅정으로 설계하였음.
- JW1601 정의 전 용량에 대한 용출 특성은 용출규격설정 가이드라인에 따른 폭넓은 pH 조건에서 기준값 이상의 용출률을 나타내며 용량별 동일한 정제 질량을 가지면서 주성분 JW1345의 비율 영향을 제제설계적으로 최소화시켜 품질적으로 차이가 적은 JW1601 정으로 개발하였음.
- Scale up 시 발생 가능한 위험요인을 파악하여 제조를 위한 공정 조건 범위를 설정하였으며 전 용량에 대한 제조 공정 조건은 Scale up 제조를 통해 평가되었고 그 결과 공정 관리기준과 품질 기준에 적합한 용출률을 나타내며 전 용량 동등하였음. 최종 조성에 대한 공정 최적화 연구를 통해 설정된 공정 조건이 재현성과 완건성이 있음을 확인하였음.
- 임상시험용 JW1601 정 그리고 위약은 제이더블유중외제약㈜에서 연구 개발하고 위탁 제조 사에 기술이전을 통해 임상약의 전 제조공정과 시험 성적 평가를 수행하였으며 임상시험을 위한 JW1601정 그리고 위약의 임상시험용 생산은 기술이전을 통해 개발된 공정조건으로 시행되었고 전 용량 확보하였음. 더불어 설정된 기준 및 시험방법에 의해 평가한 결과 모든 배치가 목표 품질 기준에 적합하였음.
- 임상시험약의 조성에 대한 안정성시험은 진행되고 있으므로 임상시험약의 차광과 흡습을 방지하기 위해 가장 안정하다고 판단되는 알루미늄/알루미늄 블리스터 포장을 임상시험용으로 우선 선정하였으며 추후 포장별 안정성시험을 통해 설정할 계획임.

마. 임상 1상 IND 승인 및 라이선스 아웃
- 임상 Protocol 및 IB를 포함한 다음의 문서들이 임상시험계획승인신청 (2018년 10월 1일).
- 식품의약품안전처로부터 임상시험진행 승인（2018년 11월 6일).
- 승인된 Protocol 제목 :건강한 성인에서 JW1601 경구 투여 시 안전성/내약성 및 약동학/약력학적 특성 평가를 위한 용량군별 무작위배정, 이중눈가림, 위약대조, 단회/ 반복투여, 단계적 증량 제 1상 임상시험 (과제 수행 :세브란스 병원)
- 세브란스병원에서 임상시험을 시행하기에 앞서, 임상시험계획서 등의 검토를 위해 세브란스병원 연구심의위원회(IRB)에 관련 문서를 제출함 (2018년 11월 6일).
- 덴마크 Leo Pharma社와 JW1601 물질용도특허 독점 라이선스 계약 체결 (2018년 8월 24일).

(출처 : 요약문 5p)

Abstract ▼

A. Preclinical study

Preclinical Toxicity
General toxicity
▶ Single-dose toxicity in rat
- Approximate lethal dose (ALD), PO: 1000 mg/kg in female, 2000 mg/kg in male
- Approximate lethal dose (ALD), IV: above 200 mg/kg
▶ 2-week repeat toxicity in rat
- Maximum tolerable dos

A. Preclinical study

Preclinical Toxicity
General toxicity
▶ Single-dose toxicity in rat
- Approximate lethal dose (ALD), PO: 1000 mg/kg in female, 2000 mg/kg in male
- Approximate lethal dose (ALD), IV: above 200 mg/kg
▶ 2-week repeat toxicity in rat
- Maximum tolerable dose (MTD): 500 mg/kg/day
▶ 4-week repeat toxicity in rat
- No observed adverse effect level (NOAEL): 250 mg/kg/day
▶ Single-dose toxicity in monkey (dose-escalating)
- Dose level selection in repeat toxicity: below 250 mg/kg
▶ 2-week repeat toxicity in monkey
- Maximum tolerable dose (MTD): 100 mg/kg/day
▶ 4-week repeat toxicity in monkey
- No observed adverse effect level (NOAEL): 80 mg/kg/day

Genotoxicity
▶ In vitro AMES study　：Negative
▶ In vivo MNT assay：Negative
▶ In vivo Comet assay : Negative

Safety pharmacology
▶ Cardiovascular safety
- hERG : IC50 > 100 uM
- Telemetry (blood pressure, rate/ECG): No effect up to 80 mg/kg
▶ Respiratory safety：Rate/respiratory function: No effect up to 500 mg/kg
▶ CNS safety：FOB: No effect up to 500 mg/kg

Method validation
▶ Method validation for formulation and evaluation
- Validated (stable for 71 hrs at room temperature, 10 days at refrigerated)
- Complete the formulation analysis for 4-week rat and monkey studies
▶ Method validation for bioanalysis and evaluation
- Validated
- Complete the bioanalysis for 4-week rat and monkey studies

Pharmaco-kinetics
In vitro ADME
▶ Caco-2 permeability：High permeability
▶ Plasma stability：Half-life value: more than 277 min in 5 species
▶ Metabolic stability in microsomes
- Metabolized in human liver microsomes with 30~40% degradation
▶ Metabolic stability in hepatocyte：Half-life value: more than 457 min in 5 species
▶ Metabolite identification：M1 metabolite in monkey and dog
▶ UGT phenotyping
- Remaining parent compound with seven UGT enzymes: more than 96%
- No UGT enzymes were involved in metabolism
▶ Plasma protein binding
- Low to moderate protein binding, no significant concentration-dependence
▶ Blood to plasma partition assay
- Distributed uniformly in blood cells and plasma in all species
▶ CYP450 inhibition
- Maximal inhibition rate for CYP1A2, CYP2B6, CYP2D6, CYP2C8, CYP2C9, CYP2C19: lower than 9%
- Maximal inhibition rate for CYP3A4: 19.72%
▶ CYP450 Induction：Not induce the enzymatic activity /CYP1A2, CYP2B6, CYP3A4 (maximum 10μM)
▶ GSH trapping study：No GSH conjugate was detected
▶ UGT inhibition enzyme test
- Inhibitory effects on activity of UGT1A9(38.8%) and UGT2B7(39.1%) at 50μM
- No effect (UGT1A1, UGT1A3) or minimal effect (UGT1A4, UGT1A6) at 0~50μM

Pharmaco-kinetics
In vivo Pharmaco-kinetics
▶ Rat pharmacokinetics
- C_max 6671±1715 ng/mL, T_max 1~2 hr, AUCT_(0-∞) 33328 ±2892 hr*ng/mL, BA 70.0 ± 6.1%
- Dose-proportional, no accumulation, no sex-differences, no food effect
▶ Monkey pharmacokinetics
- C_max 2712±401 ng/mL, T_max 0.5~2 hr, AUCT_(0-∞) 12554 ± 346 hr*ng/mL, BA 84.4 ± 2.6%
- Dose-proportional, no accumulation, no sex-differences, no food effect

Radio-labeled compound
▶ Radio-labeled ¹⁴C-JW1345, confirmed the suitability

QWBA
▶ Autoradiography/Mass balance
- Highly distributed in GI than other organs, quick elimination, low distribution in eye, brain and white fat tissue

Efficacy/Additional indication
Psoriasis
▶ 1^st evaluation/ Imiquimod-induced psoriasis-like model
- BID
- Significantly reduce the epidermal thickness (similar efficacy with positive control)
- Significantly reduce the back thickness and scaliness score
▶ 2^nd evaluation/ Imiquimod-induced psoriasis-like model
- BID and QID
- Significantly reduce the epidermal thickness and back thickness
(similar efficacy with positive control)
- Significantly reduce the scaliness score
▶ Human skin transplant psoriasis model
- Significantly reduce the epidermal ridge thickness and Ki67 index
(similar efficacy with positive control)

B. Optimization of the JW1345 synthesis process
We have studied optimization of the JW1345 synthesis process. The reactions were optimized by applying QbD (Quality by design) for each step. Also, We synthesized and purified the impurities that could be formed in the reactions. Structures of those impurities were indentified by NMR and Mass spectroscopy. We set up a fate-map about impurities and specifications.

The 6 times of scale up were produced with the process obtained through the optimization study. The JW1345(API) that obtained by scale-up were used for formulation study and non-clinical study.

JW1345(API) for clinical study was produced with JW pharmaceutical's synthesis process in cGMP site (ST Pharm). JW1345(API) was obtained 11.69 kg and was suitable all of specifications. JW1345(API) for clinical study was delivered to WuXi AppTec, which produces drug products.

C. Active Pharmaceutical Ingredient(API) and Drug Product(DP) Analytical procedure set-up & Stability data
(1) DP Analytical procedure
Test methods for JW1601 tablet
Summarizes the plans of stability studies on test batches

D. Stability test results
For the 36-month long-term stability study, the stability data of all dose levels obtained up to the 6th month have been evaluated. A significant change in the stability profile has not been identified until now. Therefore, the long-term stability study will be continued until the 36th month. Based on the successfully completed 6 months accelerated stability data, all of the test results were within their respective test specifications without any significant changes.

E. Drug product formulation study
JW1601 tablet is an investigational drug product used in the development of an oral medicinal product for the treatment of atopic dermatitis. The formulation development of JW1601 that is suitable for commercial use has been reviewed. Our development strategy of JW1601 tablet is to develop not only an immediate-release formulation to confirm the safety and efficacy of multiple dose levels, but also a stable preparation showing an equivalent dissolution pattern at different dose levels. Three dose levels, including of FR1345, were selected for a phase 1 clinical trial. The investigational drug product with the same mass at all dose levels was designed as a light pink, round, film-coated tablet, of which the appearance is similar at all dose levels.

JW1345 as the active ingredient is a sulfate hydrate form that is sparingly soluble and of which the physical properties are stable. Hence, it was designed as an excipient used generally in the development of an immediate-release formulation. Opadry an excipient in which raw materials listed in the pharmacopoeia are combined, was used as a coating agent.

Excipients were screened by effect of use and then selected. The composition of the investigational drug product per dose level was determined based on the composition profile of selected excipients. The excipients of JW1601 tablet at each dose were equal. The thickness of the tablet was similar at all dose levels by adjusting the amount of excipients depending on the percentage of JW1345 as the active ingredient. To design JW1601 tablet with three dose levels as an immediate-release formulation, the impact on the disintegration time depending on the percentage of the active ingredient was adjusted with the amount of disintegrants used. For JW1601 tablet with three dose levels, under pH conditions was established as the dissolution properties according to the guideline for the setting of dissolution specifications. Since the mass of one tablet is equal at the three dose levels, the impacts on the percentage of JW1345, an active ingredient, at the different dose levels were minimized at the time of formulation design. Therefore, there is very little difference in quality between the dose levels of JW1601 tablet.

Considering the effect of the active ingredient on the compressibility of tableting materials in the manufacturing process, the percentage of lubricants was adjusted depending on the dose level. To ensure the flowability and compressibility of tableting materials in the manufacturing process, the percentage of JW1345 as the active ingredient in terms of mass was established, with consideration for the physical properties such as the low density, and poor flowability of the active ingredient. In addition, a wet granulation process was applied.

Critical process parameters (CPPs) for each manufacturing process should be determined to satisfy the quality target profile established based on the formulation of the investigational drug product. The manufacturing process included granulation, blending, tableting, and coating processes. Risk factors that may occur in scale-up manufacturing should be investigated by identifying the effects of process parameters for each manufacturing process on the quality. In addition, the range of risk factors was established to carry out in-process control (IPC). Manufacturing process parameters were established by evaluating variables in manufacturing process as a scale-up, were evaluated. As a result, scale-up results all satisfied IPC and specification and all tablets equivalently. Process optimization studies have shown that the final formulas and manufacturing process represents robust and reproducible.

JW1601 including its placebo, and tablets was developed as the investigational drug product by JW Pharmaceutical Corporation. The whole manufacturing process and analytical outcomes of the investigational drug product were evaluated through a technology transfer from JW Pharmaceutical Corporation. The manufacture of JW1601 Tablet and Placebo batches for clinical trial study was executed based on developed procedures by technical transfer. The finished products for all batches were tested based on procedures specified in the approved analytical methods. There are no process deviations made to the testing procedure for any product. The finished products were satisfied with target quality specifications.

Stability testing for the composition of the investigational drug product is ongoing, so an aluminum-aluminum blister pack that is considered to be the most stable in the protection from light and moisture absorption was preferentially selected for the investigational drug product. Stability tests for packaging materials will later be conducted to establish the specifications for the packaging.

F. IND approval of Phase I study
To get IND approval of Phase I study, protocol and investigator’s brochure were prepared. They were ready with the help of external resource and submitted to MFDS on Oct. 1^st, 2018 with other documents required by the law. Finally, MFDS approved to conduct the clinical trial on Nov. 6, 2018. Moreover this study will be performed in Severance Hospital, Yonsei University, so the documents approved by MFDS were submitted to Severance Hospital IRB and are waiting for the review.

(source : SUMMARY 9p)