Evaluation of Acute Oral Toxicities from Paralytic Shellfish Toxins Based on a Three-level Response Surface Pathway Design

국문 초록 (Abstract)

마비성 패류독소 중독증(paralytic shellfish poisoning; PSP)은 삭시톡신과 그 유사체로 오염된 패류를 섭취했을 때 발생하며, 저림, 구토 등의 증상에서부터 근육 마비와 심각한 경우 호흡 마비로 이어져 사망에 이를 수 있다. 독 성등가계수(toxic equivalency factors; TEFs)는 다양한 마 비성 패류독소의 독성을 표준화하여 위험성을 평가하는 데 사용된다. 마비성 패류독소를 검출하기 위해 사용되던 마우스 생체 실험(mouse bioassay; MBA)에 대한 윤리적 문제가 제기되면서 고성능액체크로마토그래피와 같은 기 기 분석법으로의 전환이 시도되고 있지만, 유사체들의 적절 한 TEF를 설정하기 위해서는 여전히 동물 모델을 통한 생 체 내 독성 데이터가 필수적이다. 본 연구에서는 동물 수를 줄이면서도 신뢰할 수 있는 경구투여 독성 결과를 얻기 위 해 삼단계 반응표면-경로 (three-level RSP) 설계를 사용했다. 인증 표준 물질을 이용하여 각 독소의 초기 용량과 조정 계 수를 결정하고 시험을 진행했으며, STX.2HCl, NeoSTX, dcSTX, GTX1&4, GTX2&3, dcGTX2&3의 반수치사량 (및 TEF) 값은 각각 451.3 (1.00), 306.5 (1.47), 860.9 (0.52), 644.5 (0.70), 915.3 (0.49), 2409.3 (0.19)로 나타났다. 도출된 TEF 값은 2016년 WHO에서 권고한 TEF 값뿐만아니라, 이 전에 보고된 경구 투여 반수치사량을 기반으로 한 TEF 값 과 강한 상관관계를 보였다. 본 연구는 마비성 패류독소 뿐 만 아니라 신규 미관리 해양생물독소에 대해 적절한 TEF를 설정하는 데 있어 삼단계 반응표면경로 설계를 윤리적 우 려와 신뢰할 수 있는 독성 데이터의 필요성 사이에서 효과 적으로 균형을 맞출 수 있는 방법으로 제안한다.

다국어 초록 (Multilingual Abstract)

Paralytic Shellfish Poisoning (PSP) occurs when humans consume shellfish contaminated with saxitoxin (STX) and its derivatives. It causes symptoms ranging from numbness and nausea to severe muscle paralysis and respiratory failure. Toxic equivalency factors (TEFs) are used to standardize the toxic effects of various PSP toxins for risk assessment. Traditional detection methods, such as mouse bioassays, have been used to set the TEFs, but ethical concerns over in vivo studies have shifted the focus toward analytical methods, such as high-performance liquid chromatography. However, in vivo data are essential for establishing TEFs, particularly for emerging marine biotoxins. This study employed a three-level response surface pathway (RSP) design, which reduced the number of animals used to evaluate the median lethal dose (LD50) of STX and its derivatives. The LD50 and TEF values for STX dihydrochloride, neosaxitoxin, decarbamoylsaxitoxin, gonyautoxins 1 & 4 (GTX1&4), GTX2&3, and dcGTX2&3 were 451.3 (1.00), 306.5 (1.47), 860.9 (0.52), 644.5 (0.70), 915.3 (0.49), and 2409.3 (0.19) µg/kg, respectively. These TEFs closely aligned with the WHO recommendations and prior oral LD50 values, with Pearson correlation coefficients of 0.969 and 0.994, respectively. This study highlights the need for accurate TEF assignments for PSP toxins and new marine biotoxins, demonstrating that the three-level RSP design balances ethical concerns and provides reliable toxicity data.

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