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NATURALS 49 a Increase in Gene Expression of Collagen 200 ** 150 100 50 0 Untreated b


200 150 100 50 0


Untreated 0.1% PhytoCodine® Increase in Gene Expression of Fibronectin ** +76% +25% 50 100 d 150 0.5% +22% +53% 150 100 50 0 Untreated


0.1% 0.5% PhytoCodine®


0.1%


Increase in Protein Synthesis of Fibronectin **


* +26% +29% *** +27% 0.5% Market Reference ** +33% ** +41% 200 ** +61% * +47% ** +51% c Increase in Protein Synthesis of Collagen


0 0.1% 0.5% PhytoCodine® Untreated


0.1% 0.5% PhytoCodine®


Market Reference 0.1%


0.5%


Figure 2: Plant-derived, matrikine-like peptides activate ECM genes and protein synthesis in a dose dependent manner. In a cell culture assay, primary human dermal fibroblasts were treated for 48h with the active or a commercially available matrikine market reference. (A, B) 0.5 % of the active ingredient activated collagen gene expression by > 50 % and fibronectin gene expression by > 75 % versus untreated. (C, D) Similarly, 0.5 % of the active ingredient increased the production of the extracellular matrix proteins pro-collagen and fibronectin by 61 % and 29 %, respectively. As expected, plant-derived, matrikine-like peptides performed as good as the market reference (a commercially available, synthetic matrikine). N = 3; Mean + SEM; Student’s t-test vs. baseline; * = p < 0.05; ** = p < 0.01; *** = p < 0.001.


initiating a series of intracellular processes that finally lead to enhanced ECM production. This way, matrikines directly contribute to skin elasticity, strength and resilience. Synthetic matrikines are a success in


cosmetics. Three specific matrikines have been shown to stimulate the formation of ECM components when used in cosmetic preparations, including collagen, I, III, and VII, elastin, or fibronectin - all of which deplete considerably when skin ages.2


surprisingly, they have been used in anti- aging preparations.1,5


A novel technology to produce natural matrikines Lipoid-Kosmetik AG developed a unique technology to make natural matrikine-like peptides available for cosmetics. We systematically searched and identified matrikine-like peptide sequences from cedar nuts. In a first step, we pre-screened diverse


plant resources for criteria based on proteomics, amino acid profiles, appropriate protein amount, as well as scientific literature, and selected cedar nut as the best natural source for matrikine-like peptides. In a second step, we tested a variety of proteases (enzymes that cleave


May 2020


proteins at defined positions) and generated several unique sets of peptides. Finally, we sequenced each set and fed thousands of peptides into a database. Using a homologous search (BLAST - Basic Local Alignment Search), we identified one set containing several cedar nut peptides with strong homology to naturally occurring matrikines of our skin (Fig.1). Matrikines are small molecules that


Not


must penetrate skin layers to reach dermal fibroblasts - their point of action. Therefore, PhytoCodine®


(now referred to


as ‘the natural matrikine-like peptide’) comes with a liposomal carrier system for hydrophilic active ingredients that enhances penetration. In fact, in vivo studies using Raman spectroscopy demonstrate that penetration of model actives increased by 50% after topical application. Matrikine peptides are sensitive to


proteolytic degradation. Protease activity in the extracellular space quickly cleaves peptides, extinguishing their biological activity. Likewise, collagen and elastin are sensitive to proteolytic degradation. Therefore, the natural matrikine-like peptide contains natural eleutherosides, potent protease inhibitors from Siberian ginseng (Eleutherococcus senticosus).


In vitro activity: Activation of extracellular matrix protein synthesis We analyzed the effect of the active on gene activation and protein synthesis of two major extracellular matrix (ECM) proteins, collagen and fibronectin. Gene expression was tested by quantifying RNA transcripts using qPCR. Protein production was tested by quantifying protein release using ELISA. As a result, the active mimics the function of matrikines that naturally occur in our skin. It activates extracellular matrix gene expression and protein synthesis (Fig 2) confirming that plant-derived, matrikine-like peptides are active and functional.


In vitro activity: plant-derived, matrikine-like peptides optimize skin structure We studied the effect of plant-derived, matrikine-like peptides on skin texture by analyzing the morphology and structural integrity of skin, together with the organization and distribution of extracellular matrix proteins like elastin and collagen. As a result, the skin surface appears smoother after treatment with plant-derived, matrikine-like peptides, and the distribution of elastin, the skin’s major component of elastic fibers, is more


PERSONAL CARE NORTH AMERICA


∆ Gene Expression %


∆ Gene Expression %


∆ Protein Synthesis %


∆ Protein Synthesis %


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