Database of targeted antitumor therapy.
Most people have relatives or friends who are sick or who have already left us. In 2017, 9.6 million people are estimated to have died from the various forms of cancer. Every sixth death in the world is due to cancer, making it the second leading cause of death — second only to cardiovascular diseases.
And it is in our power to do everything possible to combine our efforts in order to increase the life expectancy of cancer patients until their full recovery, as well as improve their quality of life by increasing the selectivity and effectiveness of drugs and reducing their toxicity.
Our method is molecularly selective, targeting certain oncogenic molecules, namely the EGFR family of tyrosine kinases, which occupy a significant share in the causes of oncogenic diseases. The diagram shows the percentage of mutations in proteins of the EGFR family in non-small cell lung cancer
For several years, our laboratory developed a method for analyzing the stability of biological complexes depending on the amino acid sequence of proteins, and studying the effect of mutations on the stability of dimeric complexes. We developed a technique that allows us to determine key amino acid residues in the formation of biological complexes, developed methods for calculating the potential energy of interaction of amino acid residues, and we obtained qualitative agreement with the dissociation constant for various mutations in proteins using proteins of the Bcl-2 family as well as Nap1 proteins, Mdm2. Mdmx, P53, P63, P73, P300.
Our project is dedicated to the development of targeted therapy aimed at treating cancer caused by mutations in the EGFR tyrosine kinase family.
Our project is dedicated to the development of new simplified methods for the preparation and analysis of the affinity of antibodies to an antigen using the example of the development of antibodies to mutant proteins of the EGFR tyrosine kinase family: HER2 and HER3. Somatic mutations of ERBB2 and ERBB3 (which encode HER2 and HER3, respectively) are found in a wide range of cancers such as soft tissue cancer, lung cancer, stomach cancer, kidney cancer, ovarian cancer, bladder cancer, and breast cancer. An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses. The antibody recognizes a unique molecule of the pathogen, called an antigen, via the fragment antigen-binding (Fab) variable region.
We are going to create a database of immunoglobulins for eighteen forms of mutant oncogenic proteins of tyrosine kinases HER2 and HER3, since the response to treatment with targeted drugs in patients with EGFR mutations in tumors is much better than to treatment with standard chemotherapy drugs.
The goal of this project is to search for immunoglobulins to 18 forms of mutHER2 and mutHER3 mutant proteins that are actively involved in a wide range of oncological neoplasms so that any researcher can minimize the financial costs of synthesizing new forms of immunoglobulins, as well as reduce the cost of using research equipment , reduction of labor costs and time spent.
Antibodies with high affinity for several forms of mutant proteins simultaneously and identification of antibodies with high affinity for wild forms of HER2 and HER3 proteins will also be determined.
The result of this work will be the development of a database of immunoglobulins, each of which will be characterized by its range of affinity for the target protein.
Our database will significantly reduce the number of synthesized immunoglobulins (up to 95%) and direct the financial costs to the remainder of the entire set, as well as reduce costs for all subsequent stages of the study of the antibody-antigen pair, including reducing the time and labor required for research work in vitro, reduced use of laboratory equipment.
The obtained and selected 5% of the best molecules found will be transferred for further preclinical studies by research and pharmacological companies
The second stage involves the experimental verification of the molecules found, the synthesis of immunoglobulins, purification, verification of their affinity in vitro, and verification of their effectiveness on cell lines. Selection of the most suitable molecules. Molecule patenting.
The results of our work will allow us to prepare groups of immunoglobulins corresponding to various mutations in the transmembrane oncological proteins HER2 and HER3. Select immunoglobulins, which are characterized by the highest affinity for the selected mutant forms of proteins, excluding a huge number of inappropriate options, leaving no more than 5% of the molecules obtained for further preclinical testing.
In addition to the molecules themselves, the procedure of commercialization and transfer of the methodology for searching for small molecules for other target proteins will also be considered. Both molecules and the very method of searching and selecting small molecules, accepting orders for finding other groups of peptides, antibodies are subject to commercialization and monetization.
Why you are going to use immunoglobulins?
A characteristic feature of monoclonal antibodies is that they target specific molecules. Therefore, they are so effective in targeted (or «molecular-targeted») therapy, when we act on a specific molecule, on which the development of the disease depends. For example, trastuzumab (Herceptin), an antitumor drug based on monoclonal antibodies, has long been used in medicine. These antibodies block the receptor on the cell surface so that the receptors do not transmit a signal for division into the cell. That is, monoclonal antibodies stop tumor growth in specific diseases. The antibody itself is not universal; it is developed for a specific molecule of a specific cancer.
Benefits of targeted therapy?
Targeted therapy does not work like chemotherapy. If the mechanism is studied, if we know the key molecule responsible for the uncontrolled cell division, then we can develop an effective targeted agent — a monoclonal antibody. As for the side effects when using drugs based on monoclonal antibodies, they can occur, but in most cases, are not as serious as when treated with other chemical drugs. The reason is that the molecules that the drug acts on are needed for normal cellular functions in other organs. Therefore, some monoclonal antibodies, for example, those that are used to inhibit the growth of blood vessels in a tumor, also inhibit the growth of blood vessels in healthy organs. Therefore, it makes sense to look for the distinctive features of malignant cells from normal cells and develop targeted therapy in the application to such features.
One of the features of oncological diseases is a change in the amino acid composition of proteins, which leads to inhibition or activation of their functions, which leads to disruption of biochemical processes in the cell. Moreover, tyrosine kinases make up about 20% of all proteins that mutate during the malignant transformation of a biological cell.
The development of targeted drugs based on mono- and biclonal antibodies will allow the development of inhibitors to mutant forms of tyrosine kinases.
What is Her2?
The human epidermal growth factor receptor 2 (HER2) is a member of the family of epidermal growth factor receptors with tyrosine kinase activity. Receptor dimerization leads to autophosphorylation of tyrosine residues in the cytoplasmic domain of receptors and initiates many signaling pathways leading to cell proliferation and oncogenesis. Enhanced or overexpression of HER2 occurs in approximately 15–30% of cases of breast cancer and in 10–30% of cases of cancer of the stomach / gastroesophageal region and serves as a prognostic biomarker. Overexpression of HER2 is also observed in other cancers, such as the ovary, endometrium, bladder, lung, colon, as well as the head and neck. The introduction of HER2-directed therapy significantly affected the outcome of patients with HER2-positive breast and stomach / gastroesophageal cancer; however, the results were disappointing in other oncological diseases with excessive expression of HER2. Since one of the goals of this project is to create selective inhibitors that will mainly bind to receptors on the surface of a malignant cell, we selected mutations in the HER2 protein that can affect binding to monoclonal antibodies and also play a role in oncological diseases.
What is HER3?
Human epidermal growth factor receptor 3 ErbB3 (Her3).
HER3 is mutated in 1-3% of primary and up to 14% of metastatic ER + breast cancers. HER3 somatic mutations occur in several cancers including colon, lung, gastric, ovarian, glioblastomas. Mutant HER3-mediated oncogenic activity is dependent on HER2 and is curtailed both in vitro and in vivo using agents that either target HER3 directly or indirectly.
The increased regulation of HER3 is associated with several malignant neoplasms in which HER3 promotes tumor progression by interacting with various receptor tyrosine kinases. Studies also indicate that HER3 contributes significantly to treatment failure, mainly due to activation of PI3K / AKT, MAPK / ERK, and JAK / STAT cascade pathways. Moreover, mutations in HER3 have highlighted the role of HER3 as a direct therapeutic target. The therapeutic effect on HER3 includes the cancellation of the kinase activity of its dimerization partners using low molecular weight inhibitors (lapatinib, erlotinib, gefitinib, afatinib, neratinib) or direct targeting of its extracellular domain.
Enhanced HER3 Expression Associated with Malignant Neoplasms several types of cancer, including ovaries, breast, prostate, stomach, bladder, lung, melanoma, colorectal and squamous cell carcinoma.
RISCS and conditions:
We are a successful group of developers who created a startup company at an early stage.
The project involves several stages:
1. The first stage — creating a database is associated with a minimum level of risk, except for such as force majeure circumstances are possible such as the loss of working ability by one of the developers, natural, economic and political disasters. In case of insufficient funding, the deadlines may increase; if there is a lack of financing, this is not a condition for canceling the project
2. The second stage contains the experimental verification of selectively found immunoglobulins in vitro.
At this stage, additional time may be required to adjust and debug the selection of immunoglobulins. Our method is in good agreement with in vitro experts, however, we cannot guarantee the successful completion of experiments in vivo, on cell lines, of small animals.
3. Search for partners and patenting developed molecules
Patenting is not common for countries all over the world and is actually unique for each individual country. Also, maintaining a patent is an expensive labor-intensive process, therefore we cannot guarantee successful completion of the project starting from the third paragraph, since in this case third parties enter the game.Pre-clinical studies, determination of the best candidates, selection of the best molecules. We cannot give a 100% guarantee for cases in vivo. Also, we cannot guarantee a successful search for partners to work on conditions that suit all parties.
4.Clinical studies. We can only switch to clinical trials after successfully completing all preclinical trials.
5.Wide coverage of work results in publications, at conferences, startup presentation
6.Conclusion of contracts for the transfer of patents to research laboratories, pharmacological companies.
7.Fulfillment of orders to third parties