Collagen pentapeptide (Lys-Thr-Thr-Lys-Ser, KTTKS) and its palmitoylated derivative (pal-KTTKS) have received a great deal of attention as cosmeceutical ingredients for their anti-wrinkle effects. The objective of this study was to evaluate stability and permeability of KTTKS and pal-KTTKS in hairle...
Collagen pentapeptide (Lys-Thr-Thr-Lys-Ser, KTTKS) and its palmitoylated derivative (pal-KTTKS) have received a great deal of attention as cosmeceutical ingredients for their anti-wrinkle effects. The objective of this study was to evaluate stability and permeability of KTTKS and pal-KTTKS in hairless mouse skin. In this study, a liquid chromatography-tandem mass spectrometric method was developed for the quantification of pal-KTTKS, and used for stability and permeability studies. Stability studies were performed using skin extracts and homogenates. Both KTTKS and pal-KTTKS were rapidly degraded, but pal-KTTKS was more stable than KTTKS. When protease inhibitors were added, the stability of both compounds (KTTKS and pal-KTTKS) improved significantly. In the skin permeation study, neither KTTKS nor pal-KTTKS was detected in the receptor solution, which indicates that neither compound could permeate through the full-thickness hairless mouse skin in the experimental conditions of this study. While KTTKS was not detected in any of the skin layers (the stratum corneum, epidermis, and dermis), pal-KTTKS was observed in all skin layers: $4.2{\pm}0.7{\mu}g/cm^2$ in the stratum corneum, $2.8{\pm}0.5{\mu}g/cm^2$ in the epidermis, and $0.3{\pm}0.1{\mu}g/cm^2$ in the dermis. In conclusion, this study indicated that pal-KTTKS had greater stability and permeability than that of un-modified KTTKS, and may be a useful anti-wrinkle and anti-aging cosmeceutical agent.
Collagen pentapeptide (Lys-Thr-Thr-Lys-Ser, KTTKS) and its palmitoylated derivative (pal-KTTKS) have received a great deal of attention as cosmeceutical ingredients for their anti-wrinkle effects. The objective of this study was to evaluate stability and permeability of KTTKS and pal-KTTKS in hairless mouse skin. In this study, a liquid chromatography-tandem mass spectrometric method was developed for the quantification of pal-KTTKS, and used for stability and permeability studies. Stability studies were performed using skin extracts and homogenates. Both KTTKS and pal-KTTKS were rapidly degraded, but pal-KTTKS was more stable than KTTKS. When protease inhibitors were added, the stability of both compounds (KTTKS and pal-KTTKS) improved significantly. In the skin permeation study, neither KTTKS nor pal-KTTKS was detected in the receptor solution, which indicates that neither compound could permeate through the full-thickness hairless mouse skin in the experimental conditions of this study. While KTTKS was not detected in any of the skin layers (the stratum corneum, epidermis, and dermis), pal-KTTKS was observed in all skin layers: $4.2{\pm}0.7{\mu}g/cm^2$ in the stratum corneum, $2.8{\pm}0.5{\mu}g/cm^2$ in the epidermis, and $0.3{\pm}0.1{\mu}g/cm^2$ in the dermis. In conclusion, this study indicated that pal-KTTKS had greater stability and permeability than that of un-modified KTTKS, and may be a useful anti-wrinkle and anti-aging cosmeceutical agent.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
제안 방법
Analysis of KTTKS was performed using Acclaim 300 C18 column (2.1×150 mm, 3 μm, Dionex, Sunnyvale, CA, USA) by isocratic elution using a mobile phase consisting of 5 mM PFPA aqueous solution and acetonitrile (87:13, v/v) as described previously (Park et al., 2012).
In this study, the LC-MS/MS method for the quantification of pal-KTTKS in the skin was developed and the stability of KTTKS and pal-KTTKS in the skin of hairless mice was examined. Various types of proteolytic enzyme inhibitors were tested to prevent the degradation of KTTKS and pal-KTTKS in the skin.
Intra-day precision was determined by repeating the analysis of each QC sample three times a day. Inter-day precision and accuracy were determined by repeating this analysis on three consecutive days. Precision (expressed as the relative standard deviation) should be <15%, and accuracy should be within ± 15%.
Intra- and inter-day precision and accuracy were determined by analyzing three replicates at each of 0.5 (LLOQ), 2, 5, and 20 μg/ml of pal-KTTKS.
5, 2, 5, and 20 μg/ml. Intra-day precision was determined by repeating the analysis of each QC sample three times a day. Inter-day precision and accuracy were determined by repeating this analysis on three consecutive days.
대상 데이터
KTTKS, GHK (Gly-His-Lys) and pal-KTTKS were obtained from Peptron (Daejon, Korea). Ethylenediaminetetraacetic acid (EDTA), DL-thiorphan (TP), thimerosal (TM), phenylmeth anesulfonylfluoride (PMSF), 1,10-phenanthroline (PNT), bovine serum albumin (BSA) and ascorbic acid 6-palmitate (pal-AA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pentafluoropropionic acid (PFPA) was obtained from TCI (Tokyo, Japan).
Hairless mice were obtained from Orient Bio Inc. (Gyeonggi, Korea). Male CrlOri:SKH1-hr strain hairless mice, weighing 25 ± 3 g, were used throughout this experiment.
3). Protease inhibitors with different mechanisms (PMSF, PNT, EDTA, TM, and TP) were selected for these studies. PMSF is an inhibitor of serine proteases including trypsin and chymotrypsin.
이론/모형
The adherent fat and subcutaneous tissue were removed. Hairless mouse skin without the stratum corneum was prepared by a tape stripping method (Benson et al., 2003; Kikwai et al., 2005). The prepared mouse skin without the stratum corneum was mounted on a side-by-side diffusion cell with the epidermal layer facing the donor compartment.
성능/효과
In conclusion, this study indicates that compared to native KTTKS, pal-KTTKS has increased stability and permeability in the skin. The pal-KTTKS formulated with PMSF and PNT as protease inhibitors may be useful as an anti-wrinkle and antiaging agent in cosmeceutical products.
In the skin permeability study of KTTKS and pal-KTTKS con ducted using hairless mouse skin mounted on Franz diffusion cells, neither KTTKS nor pal-KTTKS was found in the receptor solution over an incubation period of 48 h. Therefore, it was concluded that neither KTTKS nor pal-KTTKS could permeate through full-thickness hairless mouse skin during the incubation period of these experiments. In general, peptides are known to have difficulty in permeating into or through the skin.
5 μg/ml). Therefore, it was concluded that neither KTTKS nor pal-KTTKS could permeate through full-thickness hairless mouse skin over the time period used in these experiments.
1 μg/cm2 in the dermis. Totally, 14.6% of applied pal-KTTKS was retained in the skin:8.3% in the stratum corneum, 5.6% in the epidermis, and 0.6% in the dermis.
참고문헌 (27)
Abu Samah NH Heard CM 2011 Topically applied KTTKS: a review Int J Cosmetic Sci 33 483 490
Aoki K Kajiwara M Oka T 1984 The role of bestatin-sensitive aminopeptidase, angiotensin converting enzyme and thiorphan-sensitive “enkephalinase” in the potency of enkephalins in the guinea-pig ileum Jpn J Pharmacol 36 59 65 6094902
Babu RJ Kikwai L Jaiani LT Kanikkannan N Armstrong CA Ansel JC Singh M 2004 Percutaneous absorption and anti-inflammatory effect of a substance P receptor antagonist: spantide II Pharm Res 21 108 113 14984264
Chirita R Chaimbault P Archambault J Robert I Elfakir C 2009 Development of a LC-MS/MS method to monitor palmitoyl peptides content in anti-wrinkle cosmetics Anal. Chim. Acta 641 95 100 19393372
Choi H Kim H Park J Shin E Kim D Kim S 2009 Design and efficient synthesis of novel ascorbyl conjugated peptide with high collagen biosynthesis stimulating effects Bioorg Med Chem Lett 19 2079 2082 19268583
Draelos ZD 2007 The latest cosmeceutical approaches for anti-aging J Cosmetic Dermatol 6 2 6
Foldvari M Attah-Poku S Hu J Li Q Hughes H Babiuk LA Kruger S 1998 Palmitoyl derivatives of interferon α: potential for cutaneous delivery J Pharm Sci 87 1203 1208 9758677
Gorouhi F Maibach HI 2009 Role of topical peptides in preventing or treating aged skin Int J Cosmetic Sci 31 327 345
Katayama K Seyer JM Raghow R Kang AH 1991 Regulation of extracellular matrix production by chemically synthesized subfragments of type I collagen carboxy propeptide Biochemistry 30 7097 7104 1854722
Katayama K Armendariz-Borunda J Raghow R Kang AH Seyer JM 1993 A pentapeptide from type I procollagen promotes extracellular matrix production J Biol Chem 268 9941 9944 8486721
Kikwai L Babu RJ Prado R Kolot A Armstrong CA Ansel JC Singh M 2005 In vitro and in vivo evaluation of topical formulations of Spantide II AAPS PharmSciTech 6 E565 572 16408858
Lau WM White AW Gallagher SJ Donaldson M McNaughton G Heard CM 2008 Scope and limitations of the co-drug approach to topical drug delivery Curr Pharm Des 14 794 802 18393881
Lee CH Lee KJ Chun IK Sung YG Shin YH 1994 Degradation and stabilization of methionine enkephalin and [DAla2]-methionine enkephalinamide in the corneal extracts of rabbits J Kor Pharm Sci 24 1 9
Lowry OH Rosebrough NJ Farr AL Randall RJ 1951 Protein measurement with the Folin phenol reagent J Biol Chem 193 265 275 14907713
Na DH Youn YS Park EJ Lee JM Cho OR Lee KR Lee SD Yoo SD DeLuca PP Lee KC 2004 Stability of PEGylated salmon calcitonin in nasal mucosa J Pharm Sci 93 256 261 14705183
Ogiso T Iwaki M Tanino T Yono A Ito A 2000 In vitro skin penetration and degradation of peptides and their analysis using a kinetic model Biol Pharm Bull 23 1346 1351 11085364
Park EJ Tak TH Na DH Lee KC 2010 Effect of PEGylation on stability of peptide in poly(lactide-co-glycolide) micro-spheres Arch Pharm Res 33 1111 1116 20661722
Park EJ Kim MS Choi YL Shin YH Lee HS Na DH 2012 Liquid chromatography-tandem mass spectrometry to determine the stability of collagen pentapeptide (KTTKS) in rat skin J Chromatogr B Analyt Technol Biomed Life Sci 905 113 117
Robinson LR Fitzgerald NC Doughty DG Dawes NC Berge CA Bissett DL 2005 Topical palmitoyl pentapep-tide provides improvement in photoaged human facial skin Int J Cosmetic Sci 27 155 160
Tsai WC Hsu CC Chung CY Lin MS Li SL Pang JH 2007 The pentapeptide KTTKS promoting the expressions of type I collagen and transforming growth factor-β of tendon cells J Orthop Res 25 1629 1634 17593541
※ AI-Helper는 부적절한 답변을 할 수 있습니다.