Periodic Reporting for period 2 - PEPTICAPS (Design of polyPEPTIdes diblock copolymers as emulsifiers to produce safe, controlled and reliable novel stimuli-responsive nanoCAPSules for skin care applications) Période du rapport: 2017-04-05 au 2018-10-04 Résumé du contexte et des objectifs généraux du projet PeptiCaps project aimed at producing and validating a new family of stimuli-responsive nanocapsules able to encapsulate and efficiently protect specific active ingredients for cosmetic applications based on a novel generation of universal macroemulsifiers. Different skin damaged conditions, such as allergic contact dermatitis (ACD), irritant contact dermatitis (ICD) or skin aging were considered. The development of such nanomaterials was expected to increase the lifetime of sensible cosmetic active ingredients and ensure their triggered and localized release. The European Cosmetics industry represented €76.6 billion in 2017 with exports as key activity for all sizes of companies (over 5,500 SMEs) in a sector that employs over 27,900 people. European industry has identified nanotechnology as key enabling technology for innovative product development. New nanomaterial-related products such as UV-filters in sun creams or colloidal systems in skin and hair care are already available. Nanotechnology could bring to nanocarriers specific features as triggered release, however, efforts are necessary to avoid health or environmental risks. The specific objectives of the project were: 1) to develop a novel generation of universal macroemulsifiers able to encapsulate hydrophilic or lipophilic compounds; 2) to protect and release active ingredients upon changes in pH and presence of enzymes; 3) to produce and validate cosmetic ingredients in an economic viable way; 4) to demonstrate in relevant skin models that nanocapsules could treat damaged skin; 5) to demonstrate their safety in compliance with EU regulations; 6) to develop modeling programs able to predict the risk assessment and 7) to establish a real exploitation and business plan of the technology. Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent 4 different macroemulsifiers with different stimuli responsiveness were designed: non-, pH-, enzyme- and dual-responsive. The same macroemulsifiers were used to encapsulate either hydrophilic (dipotassium glycyrrhizate or hyaluronic acid) or lipophilic actives (curcuma extract, ceramide and retinyl palmitate). Because the success of the nanoencapsulation technology, a PCT patent was submitted to protect it. Nanocapsules demonstrated the suitability to efficiently protect sensible active ingredients; retinyl palmitate degradation under UV light and oxygen was delayed from few days to several weeks. Stimuli responsiveness was also demonstrated. pH-responsive nanocapsules suffered a complete burst at pH values close to 7.4 while enzyme-responsive nanocapsules were destroyed in the presence of MMP2 or MMP9 metalloproteinases. Both macroemulsifier production and encapsulation processes were validated at pilot scale. Two competitive processes were studied for nanocapsules production: high pressure homogenizer (HPH) and tubular flow membrane contractor (TMC). Because the higher economic viability and maturity, HPH was finally selected. However, the novelty of TMC process to produce any type of nanoemulsions was demonstrated and disseminated in two articles (one published, one accepted). A Pilot Plant working under GMP (checked by means or an external audit) was established at the coordinator’s facilities. Nanocapsules penetration was studied in healthy and damaged skin models. It was demonstrated that the macroemulsifier and the encapsulated actives remained in the epidermis. None of the capsules produced showed skin irritation. The efficacy of nanocapsules in the treatment of damaged skin was demonstrated for three different skin conditions. Technology was able to reduce cellular damage by at least 30% and encapsulation increased up to 4 fold actives anti-inflammatory effect. Other actives such as ceramide presented unexpected anti-inflammatory properties. Safety of macroemulsifiers and nanocapsules was demonstrated by means of several toxicity and ecotoxicity experiments. All the new products developed were demonstrated to be safe. Characterization gaps having certain relation with nanomaterials safety were identified, such as limitations when characterizing nanomaterials dispersed in a non-aqueous media. Different protocols were proposed for size and toxicity measurements under those conditions. An in silico model now available in CORAL website was developed for toxicological prediction. The software was further adapted for the risk assessment of any type of organic/polymer based nanoparticles. 18 papers/books related with the model were published in open-access. The project was further disseminated via website and events (15 scientific conferences and 1 professional). With the aim of commercializing the technology a new brand identity was created: EMISSARY, first presented in the industrial show In-Cosmetic Global 2018. According to the business plan, the best way of exploitation of the technology was the creation of a spin-off company, EMISSARY Cosmetics, which is expected to start its activity by 2019. The company will offer on-the-shelf products and individualized encapsulation services who own their ingredient to encapsulate. Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent) The encapsulation technology developed within the PeptiCaps project demonstrated enough versatility, efficacy, safety and cost-benefit to create a spin-off company (EMISSARY Cosmetics) for the commercialisation of new generation of smart cosmetic nanocapsules in 2019. The registration of the macroemulsifier for cosmetic use (INCI Name: Methoxy PEG-125 Ethylamido Sodium Glutamate/Valine Copolymer) allows the nanocapsules to be used already. DSL and PTS are expected to launch new products including EMISSARY technology soon. Nanocapsules production was based on safe, energy- and resource efficient manufacturing system using VOC free nanoemulsification process. The ecotoxicity validation ensured that the nanocapsules will not induce contamination of soil and water resources via daily uses and under worst scenario projection. The efficacy to treat damaged skin conditions demonstrated the way in which the new technology can improve the quality of life of people affected by contact dermatitis and photo-damaged skin. Actually, the tests carried out have demonstrated that the nanocapsules could prevent their appearance if used frequently. In addition to reducing sick leaves, the absence of symptoms is important in terms of self- and social acceptance, reduction of concerns related with daily-use chemicals and promotion of outdoor activities. The versatility of the technology offers the possibility to use it in other applications such as hair or personalized cosmetics. In addition, personalized encapsulation service can open the door to new sectors (dermatology, pharmaceutical, functional foods, etc). Because consortium covered all the value chain of a cosmetic raw material the importance of collaborations between key actors is demonstrated. The strategy of close collaboration and continuous feedback followed paved the way for market launching of products based on efficacy, safety and cost-benefit in shorter R&D times. Next step would be to further improve the working methodology and collaboration using safe-by-design (SbD) approach. The identification of gaps in characterization and the proposal of standardization protocols have opened the way for future pre-normative activities that will increase European competitiveness. Their implementation of in collaboration with the EU Nano-Safety cluster will contribute towards the framework of EU nanosafety and regulatory strategies.