Design, synthesis, characterization and in vitro evaluation of new cross-linked Hyaluronic acids for Pharmaceutical, Nutraceutical and Cosmetic applications

Hyaluronic acid (HA) is an excellent biomaterial which thanks to its peculiar properties, such as biocompatibility, biodegradability, mucoadhesiveness, hydration and viscoelasticity is currently one of the most attractive polymers for many biomedical applications. Unfortunately, many of its potential applications are limited due to its short half-life as it is rapidly degraded by the hyaluronidase enzyme. In order to improve its half-life and consequently increase its performance, in this PhD thesis, native HA was modified through cross-linking reactions in which the polymer chains are stabilized by covalent bonds using natural and biocompatible aminoacids as cross-linkers agents, such as Arginine and Ornithine to overcome the potential toxicity of commonly used synthetic molecules. CDMT / NMM was used as activating agent and the structure of the new products was characterized by 1H NMR and FT-IR to confirm the occurence of the chemical modification. The morphology of the compacted and interconnected structure characterized by cages of variable size was studied by SEM. The thermal behaviour (TGA and DSC) and the rheological properties were analized and the results had shown a stable thermal profile with a rheological behaviour liquid-like and a weak-gel profile for HA-Arg and HA-Orn respectively. Swelling studies suggested a dependence on the degree of modification and pH value. Finally, the Gel Permeation Chromatography data did not reveal significant changes in molecular weight but a lower recovery value indicated that the cross-linking process has occurred. It was also demonstrated by an in vitro degradation test that the products presented an enhanced resistance profile towards enzymatic digestions therefore the cross-linking process has reduced the susceptibility of HA to the digestive action of enzymes. An evaluation of the biological profile of HA-Arg and HA-Orn was studied as possible co-adjuvants therapy for the treatment of lung inflammation. In particular, in vitro studies were performed to evaluate the release of IL-6 and IL-8, antioxidant capacity, cytotoxicity by MTS assay, and evaluation of the integrity of the epithelial barrier. The studies were performed on Calu-3 and H441 cells evaluating the treatment of HA-Arg and HA-Orn individually and in combination with sodium ascorbyl phosphate (SAP). None of the investigated treatments appeared cytotoxic and the inflammatory and antioxidant activity was promising and different based on the cell line tested. HA-Arg and HA-Orn were employed for the production of microspheres by emulsification-solvent evaporation, as potential carrier of SAP: optimization studies led to the to the realization of MSs with spherical morphology and smooth surface. An in vitro release study have demostrated that the drug was released from microspheres in controlled manner. This work has been object of the patent application N° 102019000024117 of the 16th December 2019.Hyaluronic acid (HA) is an excellent biomaterial which thanks to its peculiar properties, such as biocompatibility, biodegradability, mucoadhesiveness, hydration and viscoelasticity is currently one of the most attractive polymers for many biomedical applications. Unfortunately, many of its potential applications are limited due to its short half-life as it is rapidly degraded by the hyaluronidase enzyme. In order to improve its half-life and consequently increase its performance, in this PhD thesis, native HA was modified through cross-linking reactions in which the polymer chains are stabilized by covalent bonds using natural and biocompatible aminoacids as cross-linkers agents, such as Arginine and Ornithine to overcome the potential toxicity of commonly used synthetic molecules. CDMT / NMM was used as activating agent and the structure of the new products was characterized by 1H NMR and FT-IR to confirm the occurence of the chemical modification. The morphology of the compacted and interconnected structure characterized by cages of variable size was studied by SEM. The thermal behaviour (TGA and DSC) and the rheological properties were analized and the results had shown a stable thermal profile with a rheological behaviour liquid-like and a weak-gel profile for HA-Arg and HA-Orn respectively. Swelling studies suggested a dependence on the degree of modification and pH value. Finally, the Gel Permeation Chromatography data did not reveal significant changes in molecular weight but a lower recovery value indicated that the cross-linking process has occurred. It was also demonstrated by an in vitro degradation test that the products presented an enhanced resistance profile towards enzymatic digestions therefore the cross-linking process has reduced the susceptibility of HA to the digestive action of enzymes. An evaluation of the biological profile of HA-Arg and HA-Orn was studied as possible co-adjuvants therapy for the treatment of lung inflammation. In particular, in vitro studies were performed to evaluate the release of IL-6 and IL-8, antioxidant capacity, cytotoxicity by MTS assay, and evaluation of the integrity of the epithelial barrier. The studies were performed on Calu-3 and H441 cells evaluating the treatment of HA-Arg and HA-Orn individually and in combination with sodium ascorbyl phosphate (SAP). None of the investigated treatments appeared cytotoxic and the inflammatory and antioxidant activity was promising and different based on the cell line tested. HA-Arg and HA-Orn were employed for the production of microspheres by emulsification-solvent evaporation, as potential carrier of SAP: optimization studies led to the to the realization of MSs with spherical morphology and smooth surface. An in vitro release study have demostrated that the drug was released from microspheres in controlled manner. This work has been object of the patent application N° 102019000024117 of the 16th December 2019

Iranian Medicinal Plants: From Ethnomedicine to Actual Studies

Iran has a rich and diverse cultural heritage, consisting of a complex traditional medicine deeply rooted in the history of the territory that goes back to the Assyrian and Babylonian civilizations. The ethnomedical practices that can be identifiable nowadays derive from the experience of local people who have developed remedies against a wide range of diseases handing down the knowledge from generation to generation over the millennia. Traditional medicine practices represent an important source of inspiration in the process of the development of new drugs and therapeutic strategies. In this context, it is useful to determine the state of the art of ethnomedical studies, concerning the Iranian territory, and of scientific studies on plants used in traditional Iranian medicine. Data regarding 245 plants used in Iranian ethnomedical practices and scientific studies conducted on 89 plants collected in the Iranian territory have been reported. All of the scientific studies here reported draw inspiration from traditional medicine. The World Health Organization (WHO) has repeatedly called for an intensification of the scientific validation processes of traditional medicines intended as an important contribution to public health in various parts of the world. The process of study and validation of Iranian ethnomedical practices appears to be at an early stage

Design, Synthesis, Characterization, and In Vitro Evaluation of a New Cross-Linked Hyaluronic Acid for Pharmaceutical and Cosmetic Applications

Hyaluronic acid (HA), an excellent biomaterial with unique bio properties, is currently one of the most interesting polymers for many biomedical and cosmetic applications. However, several of its potential benefits are limited as it is rapidly degraded by hyaluronidase enzymes. To improve the half-life and consequently increase performance, native HA has been modified through cross-linking reactions with a natural and biocompatible amino acid, Ornithine, to overcome the potential toxicity commonly associated with traditional linkers. 2-chloro-dimethoxy-1,3,5- triazine/4-methylmorpholine (CDMT/NMM) was used as an activating agent. The new product (HA–Orn) was extensively characterized to confirm the chemical modification, and rheological analysis showed a gel-like profile. In vitro degradation experiments showed an improved resistance profile against enzymatic digestions. Furthermore, in vitro cytotoxicity studies were performed on lung cell lines (Calu-3 and H441), which showed no cytotoxicity

Skin Damages—Structure Activity Relationship of Benzimidazole Derivatives Bearing a 5-Membered Ring System

In the search for scaffolds for multifunctional compounds we investigated the structure activity relationship of a class of benzimidazole derivatives bearing 5-membered ring. The newly synthesized and the already known compounds were divided into three classes that present different substituent at 5 position of the benzimidazole ring (-H, -COOH or –SO3H) and different heterocycle at position 2 (thiophene, furan or pyrrole). All the derivatives were synthesized and tested to determine their photoprotective profile against UV rays, in vitro antioxidant capacity against different radicals (DPPH and FRAP test), antifungal inhibitory activity (dermatophytes and Candida albicans), antiviral and antiproliferative activity. A Structure-Activity Relationship study indicated compound 10, bearing a pyrrole heterocycle on the benzimidazole ring, as the best multifunctional derivative of the series and as potential candidate for the development of drugs especially in case of melanoma

Benzothiazole Derivatives as Multifunctional Antioxidant Agents for Skin Damage: Structure–Activity Relationship of a Scaffold Bearing a Five-Membered Ring System

Skin diseases often give multifactorial damages; therefore, the development of multifunctional compounds represents a suitable approach especially against disorders that are induced by oxidative stress. Thus, taking into account the successful results we achieved on benzimidazoles, we have devised a new series of isosteric benzothiazoles and investigated their antioxidant, photoprotective, antifungal and antiproliferative activity. Particular attention has been paid to synergistic antioxidant and photoprotective properties. For compounds 9a and 10a, a multifunctional profile was outlined, supported by an excellent filtering capacity, mainly UVB, which has higher capacities than those of the reference PBSA which is currently in the market as a UV sunscreen filter. The two compounds were also the best in terms of growth inhibition of dermatophytes and Candida albicans, and 10a also showed good antioxidant activity. Furthermore, 9a was also effective on melanoma tumor cells (SK-Mel 5), making these compounds good candidates in the development of new skin protective and preventive agents

A Safe-by-Design Approach for the Synthesis of a Novel Cross-Linked Hyaluronic Acid with Improved Biological and Physical Properties

first_pagesettingsOrder Article Reprints Open AccessArticle A Safe-by-Design Approach for the Synthesis of a Novel Cross-Linked Hyaluronic Acid with Improved Biological and Physical Properties by Sabrina Sciabica 1,†,Riccardo Barbari 1,†ORCID,Riccardo Fontana 2ORCID,Giovanni Tafuro 3,Alessandra Semenzato 4ORCID,Daniela Traini 5,6ORCID,Dina M. Silva 6,Larissa Gomes Dos Reis 6,Luisa Canilli 7,Massimo Terno 7,Peggy Marconi 2ORCID,Anna Baldisserotto 1,*ORCID,Silvia Vertuani 1,* andStefano Manfredini 1ORCID 1 Department of Life Sciences and Biotechnology, University of Ferrara, via L. Borsari 46, 44121 Ferrara, Italy 2 Department of Chemical, Pharmaceutical and Agricultural Sciences, University of Ferrara, via Fossato di Mortara 64/B, 44121 Ferrara, Italy 3 Unired Srl, via Niccolò Tommaseo 69, 35131 Padova, Italy 4 Department of Pharmaceutical and Pharmacological Sciences, University of Padova, via Marzolo 5, 35131 Padova, Italy 5 Macquarie Medical School, Faculty of Medicine, Health & Human Sciences, Macquarie University, Campus Macquarie Park, Sydney 2109, Australia 6 Woolcock Institute of Medical Research, 431 Glebe Point Road, Glebe, Sydney 2037, Australia 7 Istituto Ganassini S.p.a., Via Carlo Boncompagni, 63, 20139 Milano, Italy * Authors to whom correspondence should be addressed. † These authors contributed equally to this work. Pharmaceuticals 2023, 16(3), 431; https://doi.org/10.3390/ph16030431 Received: 20 February 2023 / Revised: 7 March 2023 / Accepted: 9 March 2023 / Published: 11 March 2023 (This article belongs to the Special Issue Novel Applications of Natural Polysaccharides for Pharmaceuticals) Download Browse Figures Versions Notes Abstract Hyaluronic acid (HA) is a polymer with unique biological properties that has gained in interest over the years, with applications in pharmaceutical, cosmetic, and biomedical fields; however, its widespread use has been limited by its short half-life. Therefore, a new cross-linked hyaluronic acid was designed and characterized using a natural and safe cross-linking agent, such as arginine methyl ester, which provided improved resistance to enzymatic action, as compared to the corresponding linear polymer. The antibacterial profile of the new derivative was shown to be effective against S. aureus and P. acnes, making it a promising candidate for use in cosmetic formulations and skin applications. Its effect on S. pneumoniae, combined with its excellent tolerability profile on lung cells, also makes this new product suitable for applications involving the respiratory tract