초록 ▼

B형간염은 전 세계 약 3.5억 명이 만성감염자이며 매년 백만 명 이상이 간경화 내지 간암으로 사망하는 매우 심각한 질병이다. B형간염을 원인으로 국내에서 약 1,100여 건의 간이식이 실시되고 있으며, B형간염 면역글로불린(HBIG)을 재발방지용으로 투여하고 있다. 현재 HBIG는 사람 혈장으로부터 분리 하고 있으나 그 원료 혈장을 전부 수입에 의존하고 있다. 이 원료혈장은 고가이며(일반혈장의 5배) 활성도가 낮고 사람의 혈액으로부터 전염되는 바이러스 등으로부터 완벽하게 안전하지 않다. 또한 수요에 탄력적으로 대처할 수 없다. 녹

B형간염은 전 세계 약 3.5억 명이 만성감염자이며 매년 백만 명 이상이 간경화 내지 간암으로 사망하는 매우 심각한 질병이다. B형간염을 원인으로 국내에서 약 1,100여 건의 간이식이 실시되고 있으며, B형간염 면역글로불린(HBIG)을 재발방지용으로 투여하고 있다. 현재 HBIG는 사람 혈장으로부터 분리 하고 있으나 그 원료 혈장을 전부 수입에 의존하고 있다. 이 원료혈장은 고가이며(일반혈장의 5배) 활성도가 낮고 사람의 혈액으로부터 전염되는 바이러스 등으로부터 완벽하게 안전하지 않다. 또한 수요에 탄력적으로 대처할 수 없다. 녹십자에서는 이러한 단점을 극복하기 위하여 유전자재조합 기술을 이용한 인간단일클론항체를 개발하여 임상1상 시험을 완료하였다. 이를 바탕으로 간이식 적응증의 상위 단계 임상시험 진행 준비를 완료하였다. 또한 간이식 후 재발방지 외에 만성간염의 치료를 위해 mouse 및 woodchuck 모델에서 예비시험을 완료하였다. 뿐만 아니라 항체의 epitope를 명확하게 규명하여 그 위치가 바이러스의 replication에 매우 중요한 위치임을 확인하였다. 이러한 결과를 바탕으로 만성간염의 치료제로서 개발에 박차를 가할 예정이다.

Abstract ▼

□ Development of an ELISA kit for rHBIG measurement
- We completed an ELISA kit for measuring rHBIG activity in human sera (Hepabig-Gene ELISA
1.1). The activity of the kit was calibrated against a reference from KFDA. This Hepabig-Gene ELISA 1.1 was compared with several commercial kits and f

□ Development of an ELISA kit for rHBIG measurement
- We completed an ELISA kit for measuring rHBIG activity in human sera (Hepabig-Gene ELISA
1.1). The activity of the kit was calibrated against a reference from KFDA. This Hepabig-Gene ELISA 1.1 was compared with several commercial kits and found that there exist correlations between different products
□ Investigation for chronic hepatitis B treatment
We established alternative animal models that can substitute chimpanzee, which is a unique animal that HBV can infect.
○ Hydrodynamic injection mouse model
- We established a mouse model that secretes HBV and HBsAg in the blood by hydrodynamic injection of HBV whole genome.
- We proved that the rHBIG could neutrlize the HBV in this model.
- We plan to see the combination of rHBIG and polymerase inhibitors in this model.
○ Woodchuck model
- We found that the rHBIG bound to the WHsAg (surface antigen of the woodchuck hepatitis virus).
- We consider to see the efficacy of rHBIG in this model.
□ Identification of epitope of rHBIG
○ 100 - 160 amino acids of HBsAg are known to be important for neutralization, which is called a major hydrophylic region (MHR).
○ 30 amino acids in the MHR were mutagenized to alanine and the rHBIG showed binding to these mutants.
○ We proved the neutralization of G145R mutant of HBsAg which is known to cause a major relapse after liver transplantation during HBIG treatment.
○ We identified the epitope of rHBIG using shotgun mutagenesis and proved that the epitope is critical for viral replication.
□ Change of clinical test plan
○ Change of a method of antibody activity measurement
- Previous method of antibody activity measurement was modified to reduce the background and the new method was validated using 500 clinical samples.
- We found out that PK profiles of clinical phase I test were changed when measured by a new method.
○ PK modeling and simulation
- We estimated doses of rHBIG to maintain the similar activity of HBIG by PK modeling and simulation (by Drs Won-Gu Kang in Youngnam Univ. & Hyung-Suk Lim in Asan Medical Center)
○ Meeting with KFDA for clinical phase II test
- There were several meetings with KFDA for clinical phase II test with the results of PK modeling and simulation but the decision was delayed due to a change of a person in charge.
- In the mean time, the contract with the CRO for clinical phase II test was completed (Oct. 2010).
- In addition, the consultation was completed with Dr. Seung-Gyu Lee in Asan Medical Center who is in charge of clinical phase II test.
○ Change of clinical test plan
- KFDA recommended to perform the additional clinical test to see the safety and pharmacodynamics of rHBIG with repeated doses in healthy volunteers although the KFDA understood the details of the changes.
- Green Cross decided to follow the recommendation of KFDA and submitted IND for clinical phase Ib test to see the safety and pharmacodynamics of rHBIG with repeated doses in healthy volunteers.
- Additional nonclinical test for 7-day repeated doses in monkeys were recommended to perform by the KFDA.
- The IND for clinical phase Ib test was approved by the KFDA on Feb. 7, 2012.
- The clinical test is scheduled to start in June, 2012 after approval of IRB.
○ Production of rHBIG for clinical test
- Green Cross completed production of rHBIG for clinical test follwing the GMP regulations.
- The total is 30,000 vials of 10,000 IU/vial and these will be used for clinical Ib and clinical II.
□ Nonclinical test for 7-day repeated doses in monkeys
○ Additional nonclinical test for 7-day repeated doses in monkeys were recommended to perform by the KFDA.
○ There were no apparent toxicities in 1.25 mg/kg infusions for 7 days in monkeys which is 3-fold excess of the maximum clinical dose.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 보고서 요약서 ... 3
• 요약문 ... 4
• SUMMARY ... 6
• 6. 연구 성과 실적 및 향후 계획 ... 8
• II. 총괄연구과제 연구결과 ... 12
• 1. 연구개발과제의 배경 및 필요성 ... 13
• 2. 국내외 기술개발 현황 ... 16
• 3. 연구개발과제의 추진체계 ... 24
• 4. 연구개발수행 내용 및 결과 ... 25
• 5. 목표달성도 및 관련분야 기여도 ... 33
• 6. 향후 연구성과 추진 계획 ... 35
• 7. 연구개발결과의 파급효과 ... 36
• 8. 연구개발결과의 활용계획 ... 38
• 9. 참고문헌 ... 38
• IV. 첨부서류 ... 41
• 끝페이지 ... 46