Overview

Our team members have a deep understanding of the suffering that diseases bring to people, having witnessed the pain and even loss of life they cause to family and friends. Additionally, many of our team members have family working in the pharmaceutical industry. As a result, they firmly believe in the importance of the medical field and have a keen interest in pharmaceutical projects, receiving strong support from their families and friends. Through brainstorming sessions and comprehensive research, we focused on cardiovascular disease (CVD), one of the leading causes of death worldwide, which claimed approximately 18 million lives in 2019, accounting for a significant portion of global mortality due to illness. Danshen (derived from the root of Salvia miltiorrhiza), a commonly used traditional Chinese medicinal herb for treating cardiovascular and cerebrovascular diseases, is our goal for producing its active ingredients using synthetic biology methods.

In all our activities, we consistently prioritize “Human Practices” as our core consideration. To promote a project that is both reasonable and responsible on a global scale, we maintain a serious and focused attitude. From the initiation of the project through each stage of its implementation, we adhere to the iGEM “Human Practices” philosophy, continuously improving the quality of our project.

Throughout the activity cycle, the work related to “Human Practices” played a significant role in every key steps. Before the project officially began, our research helped to define the feasibility direction of the project. During the experimental engineering phase, we received guidance from experts. In the discussion of the project's phased results, we enhanced the overall quality of the project through continuous optimization.

From the initial group of high school students entering the lab with a passion for synthetic biology, to walking out of the lab having accomplished the goals of the project.” The Human Practices process has taught us how to complete a responsible project for the world and continue to improve it. When conducting Human Practices, we always keep in mind the following two principles: first, to reach out to a wide range of stakeholders and to actively exchange ideas. Under the guidance of this principle, we had in-depth exchanges with botanical medicine researchers, doctors, and many other stakeholders to gain a deeper understanding of background knowledge, industry development, and project implementation. Secondly, we reflect on the lessons learned during the project and make continuous improvements. Guided by this principle, we have been able to think more about the topic by listening to a wide range of professors and experts extensively.

The Relationship between HP research and clarity process of the project direction

1.Define the Project

Literature Review

Salvianolic acid compounds are the main efficacy components of S. miltiorrhiza in activating blood circulation and removing blood stasis, with various pharmacological activities such as anti-platelet aggregation, anti-thrombosis, improving microcirculation, promoting tissue recovery, scavenging free radicals and anti-lipid peroxidation. In recent years, the demand for S. miltiorrhiza has increased due to this effect. The growth cycle of S. miltiorrhiza is up to 2 years, which will take up a large planting area. Moreover, the active ingredients of S. miltiorrhiza are low, and the productivity and utilization of the active ingredients are not high.

Rosmarinic Acid (RA) is a kind of water-soluble phenolic acid compound. It has the property of an anti-oxidant, which can prevent arteriosclerosis. Besides, it has a particular effect on anticoagulants. RA is essential for cardiovascular disease treatment. In S. miltiorrhiza, the biosynthetic pathway of phenolic acid components mainly takes 4-coumaroyl coenzyme A generated by the phenylalanine pathway as the acyl donor, and 4-hydroxyphenyllactic acid or salvinorin generated by the tyrosine pathway as the acyl acceptor, and then generates RA under the catalytic action of S. miltiorrhiza rosmarinic acid synthetase and cytochrome P450 family 98A subfamily monooxygenases (CYP98A), in that order, and further generate various complex phenolic acids. Based on the above studies, the production of RA by synthetic biology has become possible.

Among the many chassis of synthetic biology, Nicotiana benthamiana has several unique advantages, firstly, it contains a variety of biologically active components and has rich metabolic pathways for synthesizing a variety of secondary metabolites. In the synthesis, substances from tobacco itself can be directly utilized as reaction substrates for the generated products; secondly, tobacco has strong heterologous protein expression ability; Nicotiana benthamiana is also one of the earliest plants to be successfully genetically transformed, with a mature system of genetic manipulation technology.

At the level of laws and regulations, we reviewed the regulations pertaining to tobacco products, Title 5 of the Federal Food, Drug, and Cosmetic Act (FD&C).The FDA previously interpreted the exclusion in the definition of a tobacco product to mean that if a product made from or derived from tobacco is determined to have the “intended use” of a drug or device, then the product would be regulated as a medical product rather than a tobacco product. Our program ensures that all promotions and advertising of rosmarinic acid derived from tobacco are consistent and supported by scientific evidence, in accordance with FDA guidelines that address the proper labeling and advertising of drugs to prevent misleading claims. Policies in other regions, such as Asia and Europe, are more lenient.

Questionnaire

The purpose of this questionnaire was to investigate the respondents' awareness of cardiovascular health and drug therapy, as well as their attitudes and perceptions of natural ingredient drugs and genetic engineering technologies in the field of pharmaceuticals. The questionnaire covers a wide range of issues such as respondents' age, gender, health awareness, drug selection preference, and acceptance of new technologies. By analyzing the results of the questionnaire, we can understand the respondents' knowledge and concern about cardiovascular health and their attitude towards new technologies in the field of pharmaceuticals, which can provide a reference basis for research and marketing in related fields. Our goal is to increase public awareness of the benefits of using botanicals in medicine and to encourage public support for innovative health solutions.

Analysis

Through the feedback of 300 questionnaires, we got the following analysis. Through the setting of questions 1, 2, and 10, we have understood the basic information about the age, gender, and education of the participants in the questionnaire survey. As shown in the figure below, the respondents who are interested in our survey and are concerned about cardiovascular health are of all ages, especially respondents aged 41-50 and 51 years old and above. This suggests that the 40+ age group may be more concerned about cardiovascular health. This is also the age when special attention needs to be paid to understanding the disease, as some of them may be suffering from cardiovascular disease, and their opinions are crucial to our program.

According to the feedback from Questions 4 and 5, we can learn that among them, 90% have not suffered from cardiovascular and cerebrovascular diseases, but more than 60% of the respondents have relatives who have been affected by this disease. This indicates that the incidence of the disease is high, and its impact on health is significant.

From the survey feedback of question 6, we can clearly understand that people's expectation of preventive health care products (40.65%) is slightly higher than that of pharmaceuticals (39.21%). This is a good guide for us in designing our later products, which can be not only pharmaceuticals, but also some thoughts towards health care products. Questions 7, 14, and 15 let us understand that natural products are popular among the public because they are generally considered to have curative effects, and people tend to choose products containing natural ingredients for the prevention and treatment of cardiovascular and cerebrovascular diseases.

Questions 12, 16, and 17 show that there is support to produce natural products by synthetic methods (question 16, 90% support), but people do not know much about synthetic biology as well as biotechnological pharmaceuticals (the results of question 12 show that 94% of the respondents do not know what synthetic biology is; 79.14% of the respondents to question 17 do not know much about biotechnological pharmaceuticals). The above findings indicate that although most people are supportive of the use of synthetic methods to produce natural products, their knowledge of synthetic biology and biotechnological pharmaceuticals is generally low. This result shows the importance of education and awareness of new technologies.

We also conducted a survey on the ways in which people acquire information about health and medication (Question 13). As shown in the figure, more than 50% of the respondents obtain related knowledge through internet searches and social media. This indicates that the internet and social media are quite prevalent, and new technologies offer a certain convenience in information gathering. Therefore, in our dissemination of knowledge and education efforts, we will definitely prioritize the use of the internet.

Facing new technologies, people generally exhibit a cautious attitude. The feedback from questions 18, 19, 20, and 21 indicates that there are many concerns about the safety of new technology applications in pharmaceuticals. As shown in the figure, the transparency of production and sales, as well as the disclosure of information, can alleviate most people’s worries about new technologies. Of course, policy support would strengthen people’s confidence in new technologies. Regarding the biological chassis, Nicotiana benthamiana, which we intend to use, people are generally unfamiliar with it, and thus more concerned about its safety. Therefore, it is even more important for us to popularize knowledge in this area.

Interview with Professor Yi Wang

She is a researcher and doctoral supervisor at Sichuan Provincial People’s Hospital. Currently, she serves as a committee member of the Young Investigator Committee (YIC) of the International Xenotransplantation Association (IXA) (the only elected member from the Asian region), a member of the International Transplantation Society (TTS), a member of the International Xenotransplantation Association (IXA), a member of the International Pancreas and Islet Transplantation Association (IPITA), a committee member of the Youth Committee of the Clinical Pharmacy Special Committee of the Sichuan Medical Association, a standing committee member of the Clinical Pharmacy Special Committee of the Sichuan Geriatric Society, a member of the Chengdu Pharmaceutical Association, and more. She has rich experience in clinical scientific research.

In her interview, Professor Wang emphasized the clear efficacy of Salvia miltiorrhiza, particularly its role in dilating blood vessels, especially coronary arteries, while noting that it had fewer side effects. The expert's personal preference was to enhance the effectiveness of Danshen preparations by increasing the extraction rate of the active ingredient, rather than simply increasing production to avoid taking up agricultural land and increasing food imports. The experts believed that perhaps improving the existing extraction methods could be more effective in increasing the extraction rate of the active ingredients, which is crucial for the preparation of various Danshen preparations, in which a variety of secondary Danshen metabolites play a role in the therapeutic efficacy of Danshen. New methods may face the disadvantages of high research and development costs and high risk of trial and error. Prof. Wang suggested that if the extraction of active ingredients can be improved by new technologies, the application direction is broad and application scenarios can be broadened, such as nutraceuticals, oral and sublingual preparations, and skin patches used to improve local blood circulation.

Summary

Through literature review, we have learned about the existing pathway to produce rosmarinic acid in Salvia miltiorrhiza, which can be utilized to reconstruct the salvianolic acid pathway, thereby enabling the synthesis of effective components in Salvia miltiorrhiza through synthetic biology methods. By analyzing survey results and estimating the prevalence of related knowledge, we understand the general public’s concern for cardiovascular and cerebrovascular diseases, their interest in medicinal components from natural plants, but their lack of understanding of the application of plant-derived medicines and synthetic biology in drug manufacturing. Therefore, we recognize the need to strengthen science popularization efforts among the public while advancing the project, to enhance their awareness of the medicinal value of Salvia miltiorrhiza and synthetic biology technology. We enhanced the public’s understanding of the synthetic pathway of Salvia miltiorrhiza’s active components and their application in the medical field by hosting science lectures, publishing science articles, and social media campaigns. The majority of people have expressed support for our research design, considering the increase in the production of Salvia miltiorrhiza’s active components through synthetic biology methods to be innovative. Through expert interviews, we have gained insight into the necessity of the project, recognized the role of Salvia miltiorrhiza in the treatment of CVD, and proposed application scenarios we had not considered, offering valuable suggestions for the future development of the project. However, experts have also raised a significant issue regarding the research and development costs and the ultimate production efficiency. The research and development costs of drugs in the initial stage are indeed high, reminding us to pay attention to cost control in the process of project implementation. Regarding the issue of efficiency, we are preparing to conduct experiments with two synthetic pathways simultaneously: salvianic acid A + SmRAS + SmCYP98A14 and 4-hydroxyphenyllactic acid + SmRAS + SmCYP98A14 + SmCYP98A75 to produce rosmarinic acid, achieving the reconstruction of the salvianolic acid pathway, and determining the better exogenous recombination conditions by comparing the synthesis rates of rosmarinic acid under the aforementioned two operations to improve production efficiency. experts’ suggestions have guided our research direction and optimized our experimental design.

2.Execute

After the first phase, we defined the experimental strategy and designed and constructed the necessary carriers, and we continued our research on the project during the summer while doing lab experiments. We interviewed the parties involved in the project and participated in meetup events.

Interview with Professor Xing Zheng

Professor Zheng, Second Military Medical University Changhai Hospital, Department of Cardiology, Chief Physician, Professor.

Professor Zheng confirmed that he does indeed prescribe medications containing salvianolic acid components to patients in the clinical treatment of cardiovascular diseases. He mentioned that Danshen, as a traditional Chinese medicinal herb, contains salvianolic acids that are very beneficial for the treatment of cardiovascular diseases. Clinically, he often administers oral Danshen pills, Danshen tablets, as well as intravenous injections of salvianolic acid salts and Danshen injections to patients. In addition to its role in treating cardiovascular diseases, salvianolic acid salts have also been frequently used in the treatment of patients with diabetes, kidney disease, and poor renal function caused by advanced hypertension. Overall, Professor Zheng believes that Danshen-based medications offer significant assistance in clinical treatment.

Professor Zheng holds a positive view of biotechnology products and considers the use of modern biotechnology to produce pharmaceuticals to be a common and reliable method. He cited the successful synthesis of fibrinogen activators in thrombolytic therapy and monoclonal antibodies for cholesterol reduction as examples. At the same time, Professor Zheng pointed out that for traditional Chinese medicines like Danshen, biotechnology can reduce production costs, decrease land usage, and thereby improve production efficiency.

Interview with Professor Yinbiao Sun

Dr. Sun received his PhD in Pharmacy from Lund University, Sweden, and is currently working at King's College, University of London, UK. Dr. Sun is an international leader in cellular protein modification and computation/molecular imaging/protein function assays, and has extensive experience working in molecular biology laboratories.

In the interview, Prof. Sun discussed in detail the special status and advantages of Danshen in the treatment of cardiovascular diseases. Prof. Sun pointed out that Danshen, as a traditional Chinese medicine, has significant advantages in the treatment of cardiovascular diseases, and that despite its long history of use, there may be limited scope for tapping its uniqueness. Regarding the synthetic strategy we adopted, he expressed the potential of synthesizing Danshen components in Nicotiana benthamiana, especially the role in the pathway of synthesizing the active ingredient, and believed that if the active ingredient was successfully synthesized and extracted from Nicotiana benthamiana, it would be an important progress. He mentioned the high inclusion and low toxicity risk of Nicotiana benthamiana cells, and concluded that it is feasible to express Salvia divinorum components in Nicotiana benthamiana.

Prof. Sun also pointed out the possible challenges of isolating rosmarinic acid afterwards, and that even if it is successfully synthesized in tobacco, the problem of isolation still exists, and emphasized the technical difficulties of the isolation process. He also mentioned the issue of cost, to isolate the pure active ingredient at a low cost, and he also suggested that chemical synthesis may be a superior route.

To summarize, Prof. Sun's discussion covered the medical value of Salvia divinorum, the prospects of synthetic biology in drug manufacturing, and the challenges of extraction techniques, recognizing the importance of our research and the obstacles that need to be overcome.

Hangzhou Meetup

On July 27, 2024, we joined with eight other teams from Shanghai, Hangzhou, and other areas, as well as many experts and professors, at the Hangzhou Yuanzheng Qizhen Hotel for this project academic exchange event. Each team had 15 minutes to showcase their research findings, followed by a Q&A session with teachers and classmates in attendance. Our team was the third to present.

We introduced our project, which aims to achieve the low-cost synthesis of rosmarinic acid. The goal is to address the market’s insufficient supply of Salvia miltiorrhiza, ensuring the continuous production of drugs that require certain components of Salvia miltiorrhiza. We showcased our team’s work in the lab, the progress of our experiments, and the content of our interviews with experts.

During the Q&A session, Dr. Liu Shuang from Zhejiang University of Technology inquired about our experiment progress, as our project uses Nicotiana benthamiana as the chassis organism, which has a long growth cycle. This was a good reminder. In fact, we began our planting work on the first day we entered the lab and contacted experienced teachers for guidance. Additionally, the lab provided support, ensuring the smooth progress of our experiments. At the same time, we were also working on constructing vectors and other experimental content. Vice Chairman of the iGEM, Zhang Nan, asked about the inspiration for our topic, what prompted us to undertake such a project. This was indeed an important question. Undoubtedly, medicine and treating human diseases are a common interest among all team members. At the beginning of the project, through literature reading and discussions with friends and family, we determined the direction of using effective components from plants to treat major human diseases—cardiovascular and cerebrovascular diseases, with the effective components of Salvia miltiorrhiza being among the standouts.

Participating in this project sharing event involving nine teams was a rare opportunity for exchange. We not only shared our team’s progress and future plans but also learned about other synthetic biology projects, gaining inspiration from them.

3. Project Discussions

Interview with Professor Xing Zheng

In our second in-depth discussion with Dr. Zheng Xing, our focus was mainly on the products produced, especially the application considerations of these products in drug development. In addition, we discussed the challenges and market access difficulties that may be encountered if we plan to commercialize these products, i.e., develop them into marketable products. Dr. Cheng pointed out that product development must not only ensure the quality and efficacy of the drug product, but also take into account market demand, pricing strategies, competitors, and how to meet regulatory requirements, all of which are factors that must be carefully considered when successfully translating research results into marketable products.

Regarding long-term use and potential side effects, Professor Zheng believes that Salvia miltiorrhiza, as a traditional Chinese medicine, has been validated for its effectiveness through long-term use and is relatively mature in clinical applications, such as Danshen tablets and Danshen pills. Therefore, he considers Danshen to be safe. Before introducing biotechnology products into clinical use, Professor Zheng believes that the most critical safety assessment includes several steps: First, new medications need to undergo animal testing to verify their effects, from the cellular level to animal experiments, with continuous adjustments to observe the drug’s efficacy. Additionally, special attention must be paid to the experimental conditions during the drug synthesis process, such as cell culture, culture medium and cycle, as well as experimental techniques.

For clinical trials to verify the effectiveness of a product, Professor Zheng proposed the following steps:

Phase I clinical trial: conducted in normal people, mainly assessing the safety of the drug and observing whether there are any side effects.
Phase II clinical trial: conducted in patients with a smaller scope, mainly observing the efficacy of the drug, including reduction in the number of episodes, changes in electrocardiograms and changes in blood parameters compared with the control group.
Phase III clinical trials: broader in scope, randomized, and may involve multiple hospitals or even international scope to confirm the efficacy and safety of the drug before it can be applied for use.

In summary, Professor Zheng emphasized the need for rigorous animal and clinical trials to ensure the safety and efficacy of new drugs before clinical use, and was optimistic about the long-term use of Danshen.

In terms of non-clinical use in healthcare, Professor Zheng mentioned that the use of healthcare drugs in specialized medical institutions such as their tertiary hospitals was relatively rare, and that drugs such as Danshen were usually considered for symptomatic patients with symptoms such as chest tightness.

Regarding the major challenges in promoting new biotechnology products, Professor Zheng pointed out the following:

1. The need to provide data to prove the safety of new products.
2. the need to demonstrate the core competitiveness of products produced through new technologies, such as the advantages in terms of economic efficiency and drug activity.
3. Comparisons with existing drugs on the market must be made to demonstrate that the new product is superior in terms of efficiency and can reduce costs.

In summary, Professor Zheng believes that the challenges to be overcome when promoting new biotechnology products include proving the safety and affordability of the product and demonstrating its advantages over existing products.

For the overall evaluation of the project, Professor Zheng said that biomedical technology in China is advancing rapidly and that synthetic technology is a reliable method to produce pharmaceuticals. He mentioned the advantages of the project, such as saving land, increasing yield and reducing cost, and expressed his favorable opinion of the project team and encouraged us to continue our efforts.

Interview with Professor Shirui Mao

Professor Dr. Mao, Doctor of Science from Germany, PhD supervisor, Professor of Shenyang Pharmaceutical University.

She has an in-depth understanding of Salvia miltiorrhiza and pointed out that it has multiple effects such as promoting blood clot dissolution, inhibiting blood coagulation, dilating the walls of coronary arteries, improving microcirculation, promoting coronary blood circulation, reducing the incidence of myocardial infarction, and preventing arrhythmia, making it more comprehensive in treating cardiovascular diseases. Danshen has a history of over two thousand years as a medication for cardiovascular diseases, and its efficacy has been clinically verified over a long period. Currently, there are many preparations on the market with Danshen as the main ingredient, such as compound Danshen tablets, compound Danshen pills, and Guanxin Danshen capsules, which are widely used in the treatment of ischemic heart disease, hypertension, hyperlipidemia, and other cardiovascular diseases. Modern research has confirmed that Salvia miltiorrhiza contains a variety of pharmacologically active compounds, including liposoluble compounds with diterpenoid quinone structures and water-soluble compounds with phenolic acid structures, which play a key role in the treatment of cardiovascular diseases.

The application and effect of Danshen in clinical practice has received widespread attention. In response to the problem of low production and high price of Danshen, as mentioned in the last round of interview with Dr. Sun, the extraction of the active ingredient is a challenge. We address this issue by consulting Prof. Mao for solutions to active ingredient extraction. Diterpene quinones, fat-soluble compounds, and phenolic acids, water-soluble compounds, are the main active ingredients of Danshen. She mentioned the current extraction techniques, not only the traditional percolation and maceration methods, but also the emerging separation methods developed recently. As follows:

1. Ultrasonic extraction method: no need for heating, high extraction rate, fast speed, wide applicability, without destroying the structure of chemical components, but mainly used for a small number of samples extraction.
2. Supercritical extraction method (SFE): non-toxic and non-residual, low cost, safe and reliable, can be repeated recycling and regeneration of the utilization, but a single component fluid has limitations.
3. Microwave extraction (MAE): rapid heating, low energy consumption, better selectivity, but the cost of microwave equipment is high.
4. Molecular blotting technology: specificity, high efficiency, selectivity, functionality, simplicity, low cost, but higher technical complexity.
5. Enzyme extraction technology: specificity, specificity, controllability, mild reaction process, high safety, low cost, simple process, short extraction time, high extraction rate, but requires the use of specific enzymes.
6. Subcritical water extraction technology: high efficiency, low energy consumption, short extraction time, no pollution to the environment, simple equipment, easy operation, suitable for continuous extraction of different polar compounds.

Subcritical water extraction and ultrasound-enhanced-subcritical water extraction are superior to decoction, ultrasound and reflux method in terms of extraction process and extraction rate of active ingredients of Danshen, and they are more green and environmentally friendly.

Professor Mao also gave us some suggestions for the future of the product. To enhance market competitiveness, the key may lie in finding a method that can efficiently extract both the liposoluble components (such as tanshinone IIA) and the water-soluble components (such as phenolic acid compounds) from Salvia miltiorrhiza. Such a method could not only reduce production costs but also increase the utilization rate of the active ingredients, thereby enhancing the market competitiveness of the product. Currently, there is no extraction process that can efficiently extract both types of components simultaneously. Therefore, in the future, we could attempt to develop such extraction technology through research.

Interview with Professor Ying Xiao，Shanghai University of Traditional Chinese

Professor Xiao is an expert in the field of Chinese medicine research, and she has provided us with many valuable insights into the use of synthetic biology methods to extract active ingredients from Chinese herbs to develop drugs for the treatment of cardiovascular diseases.

She pointed out that the main effects of Danshen are to promote blood circulation to remove blood stasis and to clear meridians and activate collaterals, which has a significant effect on cardiovascular and cerebrovascular diseases. Current research has found that single phenolic acids, with rosmarinic acid being an important component, are significant pharmacologically active ingredients in Salvia miltiorrhiza, and their role in cardiovascular and cerebrovascular diseases is related to their skeletal structure. Although our understanding of this is limited, grasping the pharmacological knowledge is of great importance for further research. The synthesis of rosmarinic acid through synthetic biology methods has potential in the production of these active ingredients and can provide a rapid source of drugs for clinical use.

Regarding our project using Agrobacterium to transform Nicotiana benthamiana to produce rosmarinic acid, Professor Xiao, upon learning that we have basically completed the experiment, believes that our progress is quite smooth. For the production of effective medicinal components through synthetic biology, she also gave us suggestions in different directions, including the use of microorganisms such as E. coli and yeast, which can rapidly proliferate under appropriate conditions, to mass-produce target compounds; and cultivating Salvia miltiorrhiza with a higher content of salvianolic acids.

When introducing our experimental strategy, we also discussed our considerations for safety issues in the application of synthetic biology. Professor Xiao agreed and pointed out that there may be safety issues with producing salvia acid using microbial fermentation or tobacco transformation methods, similar to the controversy over genetically modified crops. Genetic engineering modifications may introduce other unknown components, posing potential risks. Being aware of the possible issues and strengthening safety awareness is important, as well as actively preventing risks. At the same time, it should be noted that countries are supportive of research in synthetic biology.

To increase yield and enzyme activity, the production process of salvianolic acid phenols can be modified through protein engineering. Synthetic biology methods can be used to synthesize other salvianolic acid substances. The key is to find the key enzymes controlling the synthesis of these metabolites and to conduct genetic modifications to increase yield. Finding the correlation between plant metabolites and gene expression can help in uncovering the genes that control key metabolic pathways. In this process, scientific research can benefit from interdisciplinary collaboration. For example, synthetic biology projects may require collaboration with structural biologists for protein crystal structure analysis and with scientists in the field of artificial intelligence for rapid gene discovery. The use of algorithms and computational methods can help narrow the scope of research and improve efficiency. Mathematical modeling and other mathematical methods are indispensable parts of scientific research. The current era offers scientists the opportunity to engage with different scientific fields, which is conducive to promoting interdisciplinary collaboration and innovative development.

Summary

After brainstorming to determine the project direction, our team members divided the tasks. Some members focused on literature research, looking for experimental methods and policy support to solve the problems. Others reached out to experts and doctors for professional guidance.

Through extensive literature research, we first identified Nicotiana benthamiana as the host cell, using the 4-coumaroyl CoA produced by the Nicotiana benthamiana's own phenylalanine pathway as the acyl donor to produce valuable salvianolic acids. Based on Professor Wang's interview and analysis of the survey questionnaire, we clarified our next steps. In terms of experiments, to enhance efficiency, we plan to validate two synthetic pathways simultaneously. Similarly, public education is a key focus to increase public awareness and support for our project.

During the summer, team members conducted experiments in the lab and interviewed multiple experts and professors, participating in relevant meetup events. They provided valuable guidance and suggestions on our experimental progress, product development, and market positioning. Our research results indicate the feasibility of producing salvianolic acids using Nicotiana benthamiana.

In subsequent discussions with professors, we obtained more valuable information. For example, Professor Xiao suggested using AI technology to optimize the structure of key enzymes in the synthetic pathway to further increase production efficiency, while Professor Mao recommended trying advanced chemical methods to improve the extraction efficiency of active components. These suggestions provide multiple optimization directions for our future work.

References

[1] Sneha Annie Sebastian MD. Cardiovascular disease risk communication: Strategies, impact, and future directions. Curr Probl Cardiol. 2024, 49(5):102490. doi: 10.1016/j.cpcardiol.2024.102490. https://www.sciencedirect.com/science/article/abs/pii/S0146280624001294?via%3Dihub
[2] Li ZM, Xu SW, Liu PQ. Salvia miltiorrhiza Burge (Danshen): a golden herbal medicine in cardiovascular therapeutics. Acta Pharmacol Sin. 2018, 39(5):802-824. doi: 10.1038/aps.2017.193.
[3] Federal Food, Drug, and Cosmetic Act (FD&C Act), United States Code, Title 21 § (2018).
[4] “The Federal Register.” Federal Register :: Request Access. Accessed July 24, 2024. https://www.federalregister.gov/documents/2017/01/09/2016-31950/clarification-of-when-products-made-or-derived-from-tobacco-are-regulated-as-drugs-devices-or.