Dr. Marie Hellfritzsch is currently working as scientist in the field of inhalation at Novartis (Sandoz Development Center). She is a pharmacist by training and completed her PhD in 2021 at Kiel University on the topic of “Zinc oxide in formulation for respiratory vaccination”. During her time as student/ PhD student she won various prizes and got the opportunity, thanks to the Aerosol Society and NordicPOP, to complete various subprojects within her PhD project abroad. Among other things, she spent a few weeks in Sydney (2016, Woolcock Institute of Medical Research), London (2018 & 2019, Kings College London) and Copenhagen (2020, University of Copenhagen). Besides her research activities, she was deeply involved in the education of students.

Project summary

The aim of my thesis with the topic “Zinc oxide (ZnO) for nasal vaccination” is to create a spray dried formulation with ZnO as an adjuvant and evoke a mucosal immune response upon nasal administration.
Immunisation via the respiratory tract is particularly promising, because antigens have their first contact with the nasal mucosa when entering the body and besides a systemic immune response, a local one is achieved, caused by the nose-associated lymphoid tissue. ZnO is well characterised in a number of clinical studies and seems to have immune adjuvant and antiviral properties. Application of spray dried formulation has a lot of advantages, e.g. no medical personnel needed, no risk of accidental needle sticks and associated infections, as well as immunisation of large population groups is possible, especially in developing countries.
In my previous work, ZnO (in various compositions) was already spray dried with mannitol (excipient, in various compositions), hyaluronic acid (adhesive) and ovalbumin (model antigen) and characterised in a pharmaceutical view. The formulation development and pharmaceutical characterisation will be followed by biopharmaceutical investigations, as it is important to see that the formulation ideas have an effect in in-vitro cell experiments and finally in-vivo. Initial in-vitro cell experiments have already been carried out with a suspension, although this does not really reflect reality, as the formulation is a powder. For this purpose, this project focuses on the work with the PreciseInhale.
Together with the working group of Ben Forbes at Kings College in London, the following work package is to be processed within 4 weeks to characterise and evaluate real-time dissolution, absorption, toxicity and transepithelial electrical resistance (TEER) of my formulations as dry powders. The project starts with the cultivation of the corresponding cell lines in order to prepare them for XposeALI. CALU-3/RPMI2650 (lung/nasal epithelial cells) will be used. As the cells grow, work continues with the Dissolvlt module. The real-time dissolution of the spray dried formulation with optical microscopy and in-vitro simulation of absorption in the lung/nasal epithelium will be investigated. The particle are deposited with this module on a glass surface and brought into contact with simulated lung/nasal mucus. Investigations are performed on the ´luminal` and the ´vascular` side. Afterwards, in-vitro cell experiments can be continued. After a defined exposure of the cells with the XposeALI module, toxicity studies and tests on TEER can be carried out.
The project aims to understand the effects of the spray dried formulations in more detail. Finally, the experiments should provide a further indication to identify an optimal formulation, which will be used for in-vivo experiments to see immunological mucosal effects. For this purpose the PreciseInhale with the ‘in-vivo module: Nose-only inhalation’ could also be used in a next step.

Career Development Award Proposal

The Career Development Grant would help me to develop professionally and personally. Since it is difficult to achieve a realistic result with conventional cell culture methods, especially in the field of dry powder formulation, this project would enable me to clarify further questions in the field of in-vitro cell experiments and to learn further techniques. Additionally the work in an intercultural working group in an interdisciplinary working environment would not only support my professional development. Ultimately, I would supplement my pharmaceutical-technological expertise with further biopharmaceutical know-how and develop myself personally.
In 2015 I finished my studies of pharmacy at Kiel University with the 2nd state examination. Inhalation is a major research focus of the Department of Pharmaceutics and Biopharmaceutics. For this reason, a focus on this topic was already set in the lectures during my studies. This aroused my interest in this area and I got the chance to spend a part of my practical year in Sydney (Australia) at the Woolcock Institute of Medical Research.
I got the chance to work on my own project with the title: "Nano- in microparticles with chitosan for nasal application" and came into contact with the topic of nasal dosage forms for the first time. Here I was already able to gain initial experience with various instruments and get to know methods playing a role in the field of nasal formulation development and characterisation. I personally have also matured further in Australia. The work taught me how to deal with setbacks and I became more self-confident, which I cannot only use during my PhD. Already during my research stay, I became aware of the fact that the field of nasal dosage forms is a rather small field of research so far.
I wanted to expand my knowledge in this field and started my PhD in the Department of Pharmaceutics and Biopharmaceutics at Kiel University in 2017. I am working in my doctoral thesis on zinc oxide as an alternative to aluminium in nasal vaccines. I started with an especially designed zinc oxide from another working group in Kiel. In their investigations, it was particularly noticeable for its antiviral and adjuvant properties. That is why pharmaceutical characterisations in comparison to alum, the mostly used adjuvant in intramuscular vaccination, were performed. Since the properties varied greatly from batch to batch and standardisation of the especially designed zinc oxide in the pharmaceutical view was impossible, I switched to normal zinc oxide as the starting material. Since zinc oxide as a raw material has particle sizes in nanoscale and thus tends to form undefined agglomerates, is too toxic in in-vitro cell experiments in high concentrations and shows a poor adsorption of proteins, the process of spray drying is used for formulation development. Finally, zinc oxide in various compositions was spray dried from a suspension with mannitol as an excipient to improve toxicity in in-vitro cell experiments and influence particle properties. Hyaluronic acid was used as an adhesive between the zinc oxide particles and as a potential penetration enhancer. Ovalbumin was used as model antigen. With the help of the methods I had already learned during my stay in Australia, the different formulations were also characterised and the methods were refined further. In order to supplement the typical characterisations with in-vitro cell experiments, we had a collaboration with Magda Swedrowska from the research group of Ben Forbes at the Kings College in London in 2018. Here I was able to learn essential skills in the field of cell culture and bring them in the cell laboratory in Kiel. A further cooperation with the working group of Ben Forbes at Kings College in London is also planned for this project to focus on the work with the PreciseInhale. Prof Ben Forbes is known as an expert in “inhalation biopharmaceutics” and has published numerous amount of research articles. "Inhalation biopharmaceutics" include anything that may affect the rate or extent of drug absorption from the lung. For research in this area, Kings College is one of the few universities with a PreciseInhale. The aim of this study is to characterise and evaluate real-time dissolution, absorption, toxicity and transepithelial electrical resistance of my formulations as dry powders. Two different modules of the PreciseInhale shall be used to expand my skills and the knowledge about my formulations with zinc oxide.
An essential part of my current work consists of student supervision and teaching. Here the knowledge acquired in the field of nasal dosage forms is passed on within the framework of small projects. In 2018, for example, two students accompanied me to Kings College in London. In addition, other areas besides nasal inhalation are taught to the students. I have already supervised four different practical courses within the pharmacy studies. I started with the practical course “compounding”, where the students learn to prepare pharmaceutical products that have to be produced within the daily routine of a pharmacy, e.g. suppositories, capsules and creams. Afterwards I supervised a course on solid dosage forms within the scope of the practical course “pharmaceutical technology”. The students learned how to make a tablet from the various starting materials in powder form to pressing the tablets with subsequent characterisation. At the moment I am working with the students in the practical course “microbiology”, where the students learn to work with microorganisms, and supervise the seminar “biopharmaceutics”. Even outside my work at the university I try to support school students in their career decisions. I am currently taking part in a “cyber mentorship program” and different courses teach me in leadership, project management and teaching skills.
During my time as a PhD student, I was able to participate in various conferences and meetings to present my work. I was at the World meeting of Pharmaceutics in Granada (2018) with a poster on: “Characterisation of Zinc Oxide Particles as Protein Carrier Systems”, at the DDL2018 in Edinburgh with a poster on: “Characterisation of Zinc Oxide as an Alternative to Aluminium Hydroxide in Nasal Vaccination” and on the Life Sciences Student Conference in Kiel (2018) I gave an oral presentation entitled “Spray dried Zinc Oxide as an Alternative to Aluminium Salts in Mucosal Vaccination”. With the latter I received a prize for my presentation. I also attended the “1st international nasal focus group meeting” in Kiel in May 2019. Numerous questions and topics were discussed and collected and it was shown, that the administration of drugs via the nose is a very broad field in which much has already been discovered, but at least as much still needs to be researched.
So my dream is to develop spray-dried formulations for nasal immunisation, to achieve immune responses comparable with alum and thus to solve some problems in the field of intramuscular immunisation, as well as to clarify existing questions of nasal drug administration.
In the long term, I would like to be involved in the field of nasal formulation and immunisation and contribute to the understanding of the nose. What is particularly important to me is that I can share my involvement and passion for my subject, both with students and at conferences.
I can imagine a career both in the form of an academic career and in the pharmaceutical industry. For me, it is important to have a broad spectrum of tasks with the possibility of cooperation. Ideally, I would like to have a smaller team in my immediate working environment, but with the opportunity to take advantage of the contacts, expertise, further education and training of a larger institution. In the future, I will pursue my work with the same passion and enthusiasm as I currently do. Of course, my future should not only be defined by my work, there should also be opportunities for a healthy work-life balance. I enjoy very much the time with my friends and my family in the nature and travel gladly to get into touch with new people and new cultures.
In the short term, I would like to share my work at conferences, enlarge my cooperation in terms of my doctoral thesis and finish my PhD next year.
The project is intended to broaden my expertise in the field of nasal dosage forms and complement my previous work with zinc oxide as a spray dried formulation.