초록 ▼

Ⅳ. 연구개발결과
본 과제수행을 통해 GLP 전임상 독성 시험용 시료(SAMiRNA-hAREG) 대량 생산을 완료하고, 불순물 분석 자료 확보하였으며, 나노입자 안정성 자료 확보를 위하여 12개월 장기보존시험, 6개월 가속시험을 통해 안정적인 나노입자를 유지함을 확인하고, 생체시료에서 정량 분석을 위한 PK 분석법을 확립하여 KIT에 기술이전을 완료하였음. Mouse Surrogate 서열 발굴을 위한 in vitro HTS을 진행하여 기존 서열대비 발현 저해 효과가 동등 이상인 신규서열을 선별하고, 동물모델에서 효능을 검증하

Ⅳ. 연구개발결과
본 과제수행을 통해 GLP 전임상 독성 시험용 시료(SAMiRNA-hAREG) 대량 생산을 완료하고, 불순물 분석 자료 확보하였으며, 나노입자 안정성 자료 확보를 위하여 12개월 장기보존시험, 6개월 가속시험을 통해 안정적인 나노입자를 유지함을 확인하고, 생체시료에서 정량 분석을 위한 PK 분석법을 확립하여 KIT에 기술이전을 완료하였음. Mouse Surrogate 서열 발굴을 위한 in vitro HTS을 진행하여 기존 서열대비 발현 저해 효과가 동등 이상인 신규서열을 선별하고, 동물모델에서 효능을 검증하였음. 전임상 독성시험은 KIT와 찰스리버에서 설치류와 영장류를 대상으로 일반독성시험, 유전독성, 안전성약리시험을 실시했으며, 그 결과 모든 파라미터에서 어떤 이상이나 변화가 발견되지 않았고, 약물동태학, 약력학, 조직병리학적으로 SAMiRNA-AREG의 안전성 입증이 확인되었음. 인간과 가장 가까운 영장류를 대상으로 한 독성 시험에서 SAMiRNA 플랫폼 기술로 개발된 치료제 후보물질인 SAMiRNA-AREG의 안전성이 입증되었다는 것에 큰 의미가 있음. IND 신청 및 승인 관련하여 경험 부족 및 의욕이 앞선 일정 설정과, COVID-19로 인한 긴급 재난상황으로 인해 임상시험용의약품(DS/DP) 제조업체 공장이 폐쇄되어 제조 일정이 다소 지연되고 있으나, CMC자료를 제외한 IND제출에 필요한 다른 자료는 완료 및 확보가 가능하므로, 임상시험 수행을 위한 치료제 신약 후보 물질 합성 생산이 완료되는 즉시 임상시험계획승인(IND) 신청을 진행할 계획이며, 성공적인 IND 승인을 위해 회사 주요 사업부에서 지속적으로 협의 및 진행해 나갈 예정임.

(출처 : 요약문 4p)

Abstract ▼

Ⅳ. Results
The SAMiRNAs with highest silencing effect on AREG were selected through extensive in vitro screening from SAMiRNA candidates designed by one-base sliding window and used for further in vivo efficacy evaluation. These studies demonstrated that :
(1) Large Scale SAMiRNA^TM

Ⅳ. Results
The SAMiRNAs with highest silencing effect on AREG were selected through extensive in vitro screening from SAMiRNA candidates designed by one-base sliding window and used for further in vivo efficacy evaluation. These studies demonstrated that :
(1) Large Scale SAMiRNA^TM Synthesis and QC evaluation for Non-clinical Tox studies : completed
(2) SAMiRNA^TM Synthesis and QC evaluation for the discovery of Surrogate Mouse SAMiRNA^TM-Amphiregulin : completed
(3) Obtained Impurity profile and Stability evaluation of SAMiRNA^TM-Amphiregulin
－ completed obtaining impurity profile
－ Through the stability evaluation(6 months of accelerated stability data and 12 months of long-term stability data), we found it had formed stable nanoparticle without any degradation or significant change in size and polydispersity over 1 year.
(4) High-throughput screening and in vivo efficacy evaluation of Surrogate Mouse SAMiRNA^TM-Amphiregulin
Administration of Amphisiran in a BLM mouse model confirmed that sequence #211 was more efficacious than sequence #39, although both sequences showed efficacy in decreasing inflammatory cells and collagen alongside ECM accumulation and edema. Ultimately, inflammatory markers and adhesion molecules were decreased when treated with SAMiRNA-AREG sequence #211.
The efficacious dose was established as 5 mg/kg intravenously.
(5) Safety and toxicity test of SAMiRNA^TM-Amphiregulin
－ Toxicology data in mice : Testing in ICR mice at weekly dose of 0, 100, 200 or 300 mg/kg Amphisiran intravenously once a week for 4 weeks (a total of 4 doses) gave no study findings of toxicological note and the No Observed Adverse Effect level (NOAEL) was determined to be 300 mg/kg/week (the maximum feasible dose). Toxicokinetic evaluation showed that systemic exposure increased with dose, was not markedly changed with repeated dosing (except for a decrease at 100 mg/kg) and there was no gender difference.
－ Toxicology data in monkey : Testing in cynomolgus monkeys at 0, 25, 50 or 100 mg/kg Amphisiran intravenously once a week for 4 weeks (a total of 4 doses)showed no study findings of toxicological note and the NOAEL was determined to be 100 mg/kg/week(the maximum feasible dose). Toxicokinetic evaluation showed that systemic exposure increased with dose, tended to decrease with repeated dosing (17-63% in males and 22-53% in females) and there was no gender difference. T1/2 in the elimination phase was 2.97-5.88 hours after the first dose and 2.44-4.01 hours after the 4th dose.
－ Genotoxicity testing : No signs of genotoxicity was seen from testing in a bacterial reverse mutation test, an in vitro chromosome aberration assay and a mouse micronucleus assay.
－ Safety Pharmacology Study : SAMiRNA-AREG did not produce any significant effects in Irwin test, Respiratory test (Respiratory Function Study), hERG Potassium Channel Preliminary Study.
－ Cardiovascular monkey telemetry Study : No effects in cynomolgus monkeys at doses of 25, 50, and 100 mg/kg and NOAEL of SAMiRNA was 100mg/kg in monkey.
(6) Documentation of SAMiRNA^TM for IND application and Approval : Ongoing These studies clearly demonstrated that SAMiRNA nanoparticles are novel siRNA platform that can be effectively and safely used for in vivo silencing of dysregulated genes in pulmonary fibrosis.

(source : SUMMARY 7p)