Volume 1 Issue 5 - September 21, 2007
Technical transfer of National Cheng Kung University biotechnology R&D result - Research and development of angiogenic proteins
Professor Hua-lin Wu

Institute of Biochemistry and Molecular Biology, National Cheng Kung University
Vascular diseases have become one of the most important diseases in the developing or developed countries worldwide. In 2002, National Cheng Kung University (NCKU) successfully obtained the “Program for Promoting University Academic Excellence” coordinated by both Ministry of Education and National Science Council and established a “Vascular Biology Group” under this program. With the full support from the Program as well as from NCKU, researchers from different universities and Academia Sinica were brought together to study the physiological, biochemical, and pharmacological aspects of vascular research. Our group from NCKU focuses on the study of biological functions of the vascular endothelial proteins. Under the financial support from the “Program for Promoting Academic Excellence of Universities“, we united physicians from the NCKU Cardiovascular Center and Division of Cardiology and Cardiovascular Surgery of NCKU Hospital to establish the Cardiovascular Research Center (CRD). The research focus of the CRC is divided into 4 major areas:
  • Cardiovascular Basic Medical Science Research
  • Stem Cell for Cardiovascular Regeneration Research
  • Clinical Drug Testing and Non-invasive Testing Device for Cardiovascular Functions Tests
  • Heart Assist Device Research
Figure 1
Part of the “Program for Promoting University Academic Excellence” resource was devoted to the research of endothelial membrane proteins, especially on the thrombomodulin (TM), a critical transmembrane protein in charge of regulating blood coagulation function. TM is a transmembrane protein whose N-terminal is located outside the membrane while the C-terminal is located inside the membrane. The head of the Nterminal is a carbohydrate-binding protein which connects to 6 epidermal growth factorlike structures (EGF) as well as a Oglycosylation site-rich domain. The Oglycosylation site-rich domain is followed by a transmembrane domain that ends with a short structure sequence inside the cell (as illustrated by Fig. 1).

The epidermal growth factor-like structures of the TM are capable of binding to thrombin. When thrombin is released in the blood vessels, it will bind to the TM on the surface of the normal vascular endothelial cells and form a complex that will change the specificity of thrombin, the blood coagulant enzyme. This complex will not only inhibit the coagulation activity of thrombin but will also promote its protein C activation capacity. As a result, protein C (APC) will be activated and in turn interfere with the important coagulation factors, Va and VIIIa which further inhibit the coagulation response. The anticoagulant function and mechanism of TM is a well-established research (as illustrated by Fig. 2).
Figure 2

During our research, two novel functions were identified for TM. First we discovered that TM is involved in the cell-cell adhesion process in that it promotes the binding of two cells and changes the morphology of the cells (see Fig. 3).
Figure 3

Through genetic engineering manipulation, a recombinant protein was produced and tested. Result showed that peptide fragment consisting of the epidermal growth factorlike structures and the O-glycosylation siterich domain of the TM (TMD23) can promote the growth and migration of cells. A further study revealed that TMD23 can enhance angiogenesis (see Fig. 4).
Figure 4

Due to the fact that angiogenesis is closely connected to many different diseases and treatments, angiogenic drugs have many practical applications in these areas. For example, angiogenesis is often observed in the formation of tumour cells. If we could block the formation of new blood vessels, we might be able to inhibit tumour growth. On the other hand, a blockade in the blood circulation will lead to cell tissue damage. With an angiogenic drug, we could repair the tissue damage by increasing blood circulation.

The TM protein developed by our research center is capable of promoting cell growth and angiogenesis and therefore has many valuable applications. Our discovery can be applied to further investigate whether TM plays an enhancing role in cancer growth or whether it is formed during angiogenesis to promote vascular formation among cancer tissues. To expand, we’d like to know whether antibodies can inhibit angiogenesis.

TM recombinant proteins and other relevant genes can be utilized to promote angiogenesis and therefore serve as angiogenic growth factors to help patients with vascular occlusion, myocardial infarction, stroke or other wound healing problems. Based on the potential of these applications, our research center filed the patent for the recombinant TM proteins and at the same time seeking for technical transfer opportunities. We were able to transfer our knowledge to Blue Blood Biotech Corp. who takes over the drug discovery responsibilities. This successful technical transfer project earned our research team a Technical Transfer Award from the National Science Council.

The R&D of a protein drug is not an easy task. There are many difficulties to overcome. First step is to understand the characteristic of the protein, including its applications, its dosage and potentiality. It is important to know what other products are already on the market and also what are the advantages of the product we are going to develop. Not until the assessment conducted yields satisfactory results will the R&D process begin. Once entered the R&D process, more tasks will surface. Not only do we need to find out the qualified mass production method, purification method and criteria. We also need to know if these proteins are stable, how to formulate these proteins into drugs, and how to store and preserve these protein drugs. Every single step has to be tested and validated. In an academic research institution, it is impossible to handle a R&D project which requires a huge amount of manpower and resources. Only a well experienced company from the industry will have the ability to complete this task. We are lucky to find Blue Blood Biotech Corp. who is willing to devote its resources in developing this protein drug. Most importantly, this R&D process is fully supported by the Ministry of Economic Affairs Industrial Development Bureau. So far Blue Blood Biotech Corp. has established a well-trained basic research foundation. They have been aggressively training their personnel. We are very confident in the collaboration between our research center and Blue Blood Biotech Corp.

What we completed so far is only a small portion of the collaboration project. What has been done is only the preparation for what lies before us. What we need to do next is to mass produce the product in a GMP regulated facility, apply for new drug R&D license, and conduct clinical trials. These are the real challenges we need to face. The costs are enormous and every step should be well planned and thought through before taken. The major reason behind this is the lack of new drug R&D experience in Taiwan where exporters are short in supply and regulations are incomplete. Because approved cases are minimal, it is highly unlikely to find any GMP facilities who will take up the job. This is the difficult situation of biotechnological drugs development commonly seen in Taiwan as well as the bottleneck of the development of biotechnological drugs. If our research results successfully enter Taiwan or US clinical trials, it should be considered an event worth celebrating for the domestic biotechnology industry.

During the progress of this project, we have gained a lot of experience on drug development and understood that new drug development is a complex process that requires a huge amount of effort and resources. In the future if Taiwanese government were to direct its industrial development towards the biotechnological medicine industry, other than building a firm basic research foundation, it is also necessary to educate and train more researchers and invest more funding or grants for R&D process. Only when a fully established integration between different resources will we to be able to succeed in biotech industry.
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