56 SKIN CARE n Placebo n 3% AnnonaSense CLR
180 160 140 120 100 80 60 40 20 0
Untreated
Capsaicin 0.1µM
Figure 3: Influence on IL-8 release..
very well organised, but only partly understood), another approach is useful. There is a strong hierarchy between different ‘biological systems’ in the human body. A well-known example of a biological system which is high in the hierarchy is our hormonal system, our so-called endocrine system. Fluctuations in hormonal status have tremendous impact on our body as everybody knows. This is for a big part the consequence of the fact that our hormones regulate many other biological systems which are lower in the hierarchy.
The ECS/EVS
The endocannabinoid (ECS) and endovanilloid systems (EVS) are other examples of biological systems which are high in the hierarchy. Where the activation of the EVS is largely detrimental, inducing inflammatory processes and an extremely important player in loss of health, the ECS can be considered to be its counterpart. The ECS is well-described in scientific literature to play an eminent role in most of our organs and tissues. The interaction between the ECS and EVS is vast. The ECS is needed to compensate for the processes induced by the EVS it seems. The intimate nature of the interaction has led to many scientists interpreting it as being one system, the ECS/EVS.
The close contact of the ECS and EVS is clearly visible in our skin. Keratinocytes, which make up more than 90% of the top layer of skin, the epidermis, show cellular receptors important for both the ECS and EVS. Of particular interest are the cellular receptors CB2 (Cannabinoid Receptor Type-2), part of the ECS, and TRPV1 (Transient receptor potential channel, vanilloid subfamily member 1), part of the EVS. The TRPV1 can be triggered by heat, UV-radiation, free radicals and pH shifts, among others. Activation of TRPV1 triggers the production of inflammatory mediators which are important in skin inflammation. Activation of TRPV1 on sensory neurons,
PERSONAL CARE ASIA PACIFIC 0.005% 0.05%
AnnonaSense CLR + Capsaicin 0.1µM
70 60 50 40 30 20 10 0
250 Hz 5 Hz
Figure 4: Assessment of skin sensitivity.
the cells which make up the sensory nerves in our skin, can lead to itch, pain and burning sensations. TRPV1 is well-described to be one of the key instigators of loss of skin health and wellbeing. Like TRPV1, CB2 is a cellular receptor which can be found both on keratinocytes and on sensory neurons in the skin. The ECS seems designed to keep the EVS under control and to establish and maintain a healthy situation. Nature has found a tremendously elegant way of managing this through the action of CB2. As CB2 is a cellular receptor, it can be activated by molecules produced in the skin or by foreign substances. Endocannabinoids, produced in our body, can activate CB2. This leads to a reaction in the human cell which, on its turn, leads to the deactivation of TRPV1. This literally restores and maintains a healthy homeostasis. In support of the endocannabinoids in our skin, activation of CB2 with a cosmetic active ingredient is therefore a potent approach in obtaining and maintaining skin health and wellbeing. CLR Berlin has developed AnnonaSense CLR (“Cherimoya”, INCI: Annona Cherimola Fruit Extract) to sustainably achieve skin health and wellbeing by interaction with CB2.
Efficacy studies In vitro assays Below, the results of the studies relevant to CB2 and TRPV1 are presented in detail. Other in vitro studies were performed additionally. It was shown that Cherimoya reduced the production of histamine- induced production of Calcitonin gene- related peptide (CGRP) and the production of Interleukin-6 (IL-6) after application of Tumor Necrosis Factorα (TNFα) and Substance P. These studies were performed on primary human keratinocytes. Another study on these cells was performed to determine the activity of Cherimoya on the production of the Interleukin-31 receptor (IL-31RA). Here it was shown that
Cherimoya could strongly reduce the production of IL-31RA, further underlining its importance for reducing skin discomfort.
Agonistic effect of Cherimoya on CB2 293T-CB2 cells (CB2-expressing human epidermal keratinocytes) were incubated with different concentrations of Cherimoya, respectively a positive control (WIN55, 212- 2 [1 μM]) for 10 minutes. The positive control is a specific agonist for CB2 and can, therefore, induce specific cellular processes induced by CB2. Subsequently the cells were treated with Forskolin (10 μM) for 6 hours. Treatment with Forskolin led to an increase in the production of cyclic adenosine monophosphate (cAMP). Interaction with CB2 by the positive control substance and the different concentrations of Cherimoya had an impact on the Forskolin-induced cAMP production. CB2- induced reduction of Forskolin-induced cAMP production was determined, where the result obtained with the positive control was set at 100%.
Results At two different concentrations, treatment with Cherimoya showed a strong agonistic effect on the CB2 receptor, where the higher concentration of the two, 0.05%, was even able to outperform the positive control, a specific agonist for CB2 (Fig 1).
Reduction of TRPV1-induced inflammation, co-cultivation of keratinocytes – sensory neurons
cAMP is needed for TRPV1-activity. Mechanistically, cAMP is needed for Protein Kinase A (PKA) and PKA-induced phosphorylation of TRPV1 is a key event in the activation of TRPV1. CB2-induced reduction of cAMP, as shown in Figure 1, is therefore a potent approach towards deactivation of TRPV1. A CB2-induced reduction of the activity of TRPV1 can elegantly be proven by making use of capsaicin. Capsaicin is a specific agonist for
September 2019
IL-8 expression (%)
Reduction of skin sensitivity (%)
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