분광광도법을 이용한 간단한 in Chemico 피부감작성 시험법의 개발
A Simple Spectrophotometric Test to Identify Skin Sensitizers and Non-Sensitizers in Chemico
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
For the key event No. 1 in skin sensitization, a hapten should react with endogenous proteins for the initiation of immune reaction. Although Direct Peptide Reactivity Assay (DPRA) has been developed as an alternative method for key event 1, artificially synthesized peptides and HPLC should be employed in performance, which are somewhat inconvenient. In the present study, a simple spectrophotometrical method to identify skin sensitizers in chemico was developed by using cysteamine, an endogenous substance that contains amino and thiol groups. To quantitate remaining cysteamine following the reaction with skin sensitizers, 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB) was selected for the derivatization. The conditions optimized included: incubation time of cysteamine with test substances, molar ratios of cysteamine to test substances, stability of derivatized products of test substances with optimal concentration and incubation time of DTNB, and spectrophotometrical characteristics of the derivatized product. With the optimized conditions, 11 skin sensitizers and non-sensitizers were tested to know whether the method is feasible or not. Although further optimization is required, it would be a useful screening tool for determining skin sensitization potential of small molecules, because the present method employs simple endogenous cysteamine as an acceptor for sensitizers with a spectrophotometric detection system.
Abstract
For the key event No. 1 in skin sensitization, a hapten should react with endogenous proteins for the initiation of immune reaction. Although Direct Peptide Reactivity Assay (DPRA) has been developed as an alternative method for key event 1, artificially synthesized peptides and HPLC should be employed in performance, which are somewhat inconvenient. In the present study, a simple spectrophotometrical method to identify skin sensitizers in chemico was developed by using cysteamine, an endogenous substance that contains amino and thiol groups. To quantitate remaining cysteamine following the reaction with skin sensitizers, 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB) was selected for the derivatization. The conditions optimized included: incubation time of cysteamine with test substances, molar ratios of cysteamine to test substances, stability of derivatized products of test substances with optimal concentration and incubation time of DTNB, and spectrophotometrical characteristics of the derivatized product. With the optimized conditions, 11 skin sensitizers and non-sensitizers were tested to know whether the method is feasible or not. Although further optimization is required, it would be a useful screening tool for determining skin sensitization potential of small molecules, because the present method employs simple endogenous cysteamine as an acceptor for sensitizers with a spectrophotometric detection system.
참고문헌 (24)
- Ahlfors SR, "Reactivity of contact allergenic haptens to amino acid residues in a model carrier peptide, and characterization of formed peptide-hapten adducts", Skin Pharmacology and Physiology, 16, pp.59~68 (2003)
- Anderson SE, "The LLNA : A brief review of recent advances and limitations", Journal of Allergy, 2011, pp.1~10 (2011)
- Autelitano A, "The reaction of cinnamaldehyde and cinnam(o)yl derivatives with thiols", Acta Pharmaceutica Sinica B, 7, pp.523~526 (2017)
- Basketter DA, "Use of the local lymph node assay for the estimation of relative contact allergenic potency", Contact Dermatitis, 42, pp.344~348 (2000)
- Basketter DA, "A comparison of statistical approaches to the derivation of EC3 values from local lymph node assay dose responses", Journal of Applied Toxicology, 19, pp.261~266 (1999)
- Basketter DA, "Comparison of the local lymph node assay with the guinea-pig maximization test for the detection of a range of contact allergens", Food and Chemical Toxicology, 30, pp.65~69 (1992)
- Cho SA, "High-throughput screening (HTS)-based spectrophotometric direct peptide reactivity assay (Spectro-DPRA) to predict human skin sensitization potential", Toxicology Letters, 314, pp.27~36 (2019)
- Cockshott A, "The local lymph node assay in practice: a current regulatory perspective", Human & Experimental Toxicology, 25, pp.387~394 (2006)
- Dimitrov S, "Accounting for data variability, a key factor in in vivo", in vitro relationships: application to the skin sensitization potency (in vivo LLNA versus in vitro DPRA) example, 2016, pp.36, 1568~1578
- Gerberick GF, "Development of a peptide reactivity assay for screening contact allergens", Toxicological Sciences, 81, pp.332~343 (2004)
- Gerberick GF, "Local lymph node assay (LLNA) for detection of sensitization capacity of chemicals", Methods, 41, pp.54~60 (2007)
- Gerberick GF, "Quantification of chemical peptide reactivity for screening contact allergens: A classification tree model approach", Toxicological Sciences, 97, pp.417~427 (2007)
- Gerberick GF, "Investigation of peptide reactivity of pro-hapten skin sensitizers using a peroxidase-peroxide oxidation system", Toxicological Sciences, 112(1), pp.164~174 (2009)
- Jowsey IR, "A future approach to measuring relative skin sensitising potency: a proposal", Journal of Applied Toxicology, 26, pp.341~350 (2006)
- Kimber I, "The local lymph node assay: past, present and future", Contact Dermatitis, 47, pp.315~328 (2002)
- Kimber I, "Cytokines and chemokines in the initiation and regulation of epidermal Langerhans cell mobilization", British Journal of Dermatology, 142, pp.401~412 (2000)
- Kimber I, "Assessment of the skin sensitization potential of topical medicaments using the local lymph node assay: An interlaboratory evaluation", Journal of Toxicology and Environmental Health, 53, pp.563~579 (2010)
- Lepoittevin JP, "Allergic contact dermatitis caused by naturally occurring quinones", Pharmaceutisch Weekblad, 13(3), pp.119~122 (1991)
- Nepal MR, "Identification of pre- and pro-haptens with a β-galactosidase-expressing E. coli cell culture system for skin sensitization", Toxicology Letters, 305, pp.81~93 (2019)
- Nepal MR, "A simple in chemico method for testing skin sensitizing potential of chemicals using small endogenous molecules", Toxicology Letters, 289, pp.75~85 (2018)
- Patlewicz G, "Can currently available non-animal methods detect pre- and pro-haptens relevant for skin sensitization?", Regulatory Toxicology and Pharmacology, 82, pp.147~155 (2016)
- Ryan CA, "Approaches for the development of cell-based in vitro methods for contact sensitization", Toxicology in Vitro, 15, pp.43~55 (2001)
- Ryan CA, "Interactions of contact allergens with dendritic cells: Opportunities and challenges for the development of novel approaches to hazard assessment", Toxicological Sciences, 88, pp.4~11 (2005)
- Urbisch D, "Peptide reactivity associated with skin sensitization: The QSAR Toolbox and TIMES compared to the DPRA", Toxicology in Vitro, 34, pp.194~203 (2016)