76 PEPTIDES


TABLE 2: BCOP TEST RESULTS (MEAN±SD). OD VALUE REPRESENTS MEAN PERMEABILITY VALUE. IVS = IN VITRO SCORE. THE UN GHS GUIDELINES ARE FOLLOWED FOR THE CLASSIFICATION


Group Negative control Positive control Test sample


Mean corrected opacity value 0.427 ± 1.7154


32.9298 ± 0.450 -1.125 ± 0.2586


of which contributed to an overall advancement in facial skin condition. Impressively, none of the participants encountered any discomfort throughout the entire 28-day testing period.


Skin irritation evaluation A comprehensive skin irritation test (Report No. HK-R-20230609-02) was commissioned following the guidelines in ‘In Vitro Skin Irritation: Reconstructed Human Epidermis Test OECD TG439’.


The assessment was executed using the


laboratory method “Skin Model Irritation Test- MTT+IL-1a”, which entails a comparative analysis against a positive control to ascertain the potential irritative impact of the test substance on the reconstituted epidermal model in vitro. The irritation induced by the test sample (T-EGF at 100 µg/ml) is determined by quantifying the percentage reduction in cellular viability. The results indicate no irritating substance


is present in the tested sample. The cell viability was determined to be 97.89% upon application of the sample, surpassing the threshold of 50%. The interleukin 1a (IL-1a) concentration was measured at 8.646 pg/mL (Table 1).


Ex vivo eye irritation test The eye irritation test of T-EGF, according to the OECD TG437 bovine corneal turbidity and permeability (BCOP) guidelines, evaluated the potential irritation of T-EGF at a concentration of 100 µg/mL to the eyes. Ultrapure water was used as negative control, and absolute ethanol was used as positive control. The results are shown in Table 2, concluding that T-EGF is classified as non-irritating.


Application of T-EGF in skincare formulations The T-EGF raw material is a white solid substance obtained through freeze-drying. It can be dissolved into diverse aqueous solutions before adding to the production batch. Its versatility allows for incorporation into various cosmetic formulations, particularly suited for repair and sensitive skin. T-EGF exhibits potency at very low


concentrations. Guidelines regarding its optimal inclusion in cosmetic preparations are detailed in Table 3.


Best practices for T-EGF Empirical studies revealed several factors that influence the stability of T-EGF formulations. Some key recommendations when applying T-EGF during production are listed below: ■ Add T-EGF to the production batch when temperature is below 45°C


PERSONAL CARE November 2023 General production guidelines Applications (final dosage)


Mean corrected OD value 0.0678 ± 0.0056


1.2279 ± 0.4143 0.0013 ± 0.0023 IVS 1.444 ± 1.7918 51.3488 ± 5.8910 -1.1064 ± 0.2926


UN GHS classification No category


No stand-alone


prediction can be made No category


Irritant classification Non-irritant


Impossible to predict Non-irritant


TABLE 3: T-EGF BASIC PRODUCT INFORMATION AND DOSAGE RECOMMENDATIONS Product name


T-EGF INCI name (T-EGF) INCI name of the complete powder Storage conditions sr-(Hexapeptide-40 Oligopeptide-232 sh-Oligopeptide-1


Sodium Chloride Disodium phosphate


sr-(Hexapeptide-40 Oligopeptide-232 sh-Oligopeptide-1) Sodium Phosphate


The freeze-dried powder is protected and can be stored in room temperature (detailed updates expected in Q4 2023)


Use a T-EGF solution of 100µg/ml at 0.2%-20% in the final product


Serum (5-10 µg/ml); Sheet masks, patches (0.2-2 µg/ml); Day and night cream (3-5 µg/ml); Lotion (1-2 µg/ml)


■ Avoid using protein deactivating substances alongside T-EGF ■ Optimal pH range for T-EGF is 4-5.5 or 6-8.5 ■ Combining with other peptides and moisturizing agents can enhance T-EGF activity ■ Energy-generating substances can enhance T-EGF efficacy


Conclusion Overall, the development of TD1 presents a novel solution in the skincare industry for transdermal delivery of macromolecules. T-EGF stands as a testament to the remarkable possibilities that arise from deeply understanding and harnessing the intricacies of the human body’s natural processes. As research and technology continue to


evolve, we expect significant momentum and interest from the industry and anticipate the emergence of more skincare formulations that benefit from inclusion of novel ingredient such as the transdermal epidermal growth factor.


whitening /anti-aging effects. Doctoral dissertation, Jiangnan University. 2008


6. Schlessinger J, Schreiber AB, Levi A, Lax I, Libermann T, Yarden Y. Regulation of cell proliferation by epidermal growth factor. CRC Critical Reviews in Biochemistry. 1983; 14:93-111


7. Xing XJ, Yang L, You Y, Zhong BY, Song QH, Deng J, Hao F. Study of the biological function and penetration pathways of the mouse epidermal growth factor ethosomal delivery system. Exp. Dermatol. 2011; 20:945-947


8. Kalluri H, Banga AK. Transdermal delivery of proteins. AAPS PharmSciTech. 2011; 12:431-441


9. Subedi RK, Oh SY, Chun MK, Choi HK. Recent advances in transdermal drug delivery. Arch. Pharm. Res. 2010; 33: 339-351


PC


References 1. Ruan R. Study on peptide mediating hEGF delivery across skin and its applications. Doctoral dissertation, University of Science and Technology of China. 2015


2. Cohen S, Elliott GW. The stimulation of epidermal keratinization by a protein isolated from the submaxillary gland of the mouse. The Journal of Investigative Dermatology. 1989; 92:157S; discussion 158S-159S


3. Pastor JC, Calonge M. Epidermal growth factor and corneal wound healing. A multicenter study. Cornea. 1992; 11:311-314


4. Yanhong T, Quanxiang W, Zaiming T. The protective effect of human epidermal growth factor on the skin. Chin. J. Med. Aesth. & Cosmet. 2004; 10(1), 3


5. Liqiong Chen. Study of hEGF in cosmetics


10. Karande P, Jain A, Mitragotri S. Discovery of transdermal penetration enhancers by high-throughput screening. Nat. Biotechnol. 2004; 22:192-197


11. Barry BW. Breaching the skin’s barrier to drugs. Nat. Biotechnol. 2004; 22:165-167


12. Paudel KS, Milewski M, Swadley CL, Brogden NK, Ghosh P, Stinchcomb AL. Challenges and opportunities in dermal/ transdermal delivery. Therapeutic Delivery. 2010; 1:109-131


13. Chen Y, Shen Y, Guo X, Zhang C, Yang W et al. Transdermal protein delivery by a coadministered peptide identified via phage display. Nat. Biotechnol. 2006; 24: 455-460


14. Prausnitz MR. A peptide chaperone for transdermal drug delivery. Nat. Biotechnol. 2006; 24:416-417


15. Ruan RQ, Wang SS, Wang CL et al. Transdermal delivery of human epidermal growth factor facilitated by a peptide chaperon. European Journal of Medicinal Chemistry. 2013; 62(62C), 405-409


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