Insulin artificial synthesis, innovation must also be synthesized in the future

At the just-concluded 2021 National Two Sessions, Ding Kuiling, a member of the National Committee of the Chinese People’s Political Consultative Conference, an academician of the Chinese Academy of Sciences, and the executive vice president of Shanghai Jiaotong University, put forward the proposal of “Being a Hot and Cold Bench for Basic Research” and received widespread attention.

This academician with a background in basic research believes that my country’s technological innovation still has an impetuous mentality that emphasizes short-term benefits and dilutes long-term effects, and that leading, disruptive, and original major scientific and technological achievements are insufficient; “high-tech” scientific and technological talents exist Larger supply gap.

Ding Kuiling’s suggestion is not unrelated to the Shanghai Institute of Organic Chemistry of the Chinese Academy of Sciences (hereinafter referred to as the “Institute of Organic Chemistry”), where he worked for a long time in the past. As early as 56 years ago, in September 1965, the Institute of Organic Chemistry, the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (hereinafter referred to as the “Institute of Biochemistry”), and Peking University made joint efforts in the field of basic research. The achievement-synthetic bovine insulin.

On September 17, 1965, the Chinese scientific research team observed the crystallization of artificially synthesized bovine insulin, and everyone cried and screamed and laughed excitedly. This means that for the first time in the world, a protein with the same chemical structure and complete biological activity as the natural insulin molecule has been artificially synthesized for the first time in the world.

Afterwards, famous scientists from the United States, Britain, France, Italy, the Netherlands, Belgium, Norway and other countries sent congratulatory messages. The United States is shocked by my synthetic bovine insulin” article. Until 1982, 17 years after the synthetic bovine insulin, British Prime Minister Thatcher made a special call to visit the Institute of Biochemistry.

“The study that I participated in at the age of 30 until now is 88 years old, there are still so many people who can remember what happened to us.” Xu Jiecheng, a retired researcher at the Institute of Organic Chemistry, is now a “big baby”, he is the whole The only one who is still alive among the experts involved in the research of synthetic bovine insulin by the Institute of Organic Chemistry. He told a reporter from China Youth Daily and China Youth Daily that synthetic bovine insulin seems to be a relatively large scientific achievement, but the scientific spirit behind it has lasted forever. This scientific spirit is especially precious to the “Innovative China” in the process of progress.

Back in 1958, when New China was founded less than 10 years ago, economic development and material conditions were relatively backward. The British chemist Sanger won the Nobel Prize in Chemistry this year for completing all the sequencing work of insulin. The academic journal Nature published a commentary that year, saying that “synthesis of insulin will be a distant thing.” On the other side of the world, in a developing country in China, in a small room of the Shanghai Institute of Biochemistry, an older Wang Yinglai, the director of the Institute of Biochemistry, takes the lead. Eight young people in their 30s are gathering together to discuss “what to study.” .

This is a “dream team” in the field of Chinese biochemistry. Wang Yinglai is a PhD in Biochemistry from the University of Cambridge, England, enzyme chemist Zou Chenglu is from Cambridge, protein expert Cao Tianqin is from Cambridge, nucleic acid expert Wang Debao is a postdoctoral fellow at Johns Hopkins University in the United States, vitamin expert Zhang Youduan is also from Cambridge, protein expert Niu Jingyi is from the United States Zhou Guangyu, a biochemical expert from Texas State University, is a Ph.D. from the University of Leuven in Belgium, Zhaowen Shen is a Ph.D. from the University of Toronto in Canada, and Jinghua Xu graduated from the Department of Chemistry at Southwest Associated University.

In this nine-person meeting, someone proposed the idea of ​​marching into life sciences and artificially synthesizing proteins, which sparked heated discussions. What scholars from developed countries in Europe and the United States do not know is that Chinese scientists with relatively “poor” scientific research facilities dare to challenge the “synthetic insulin” task that seemed impossible at the time. On December 18, 1958, the Institute of Biochemistry formally confirmed the synthetic insulin project and invited Peking University and other units to participate in the research.

At that time, Shanghai lacked talents to study peptide synthesis, and there was a serious shortage of protein scientific research materials across the country. The country can only produce low-purity sweet, refined, and gluten amino acids. The other 14 amino acids used to synthesize insulin have to be re-exported from Hong Kong, which is expensive. But the innovative ideas of scientists are not constrained by material difficulties.

The Institute of Biochemistry officially confirmed the synthetic insulin project. At the same time, it explored and built a Dongfeng Biochemical Reagent Factory specializing in the preparation of reagents such as amino acids to provide a source of amino acids for the synthetic insulin project. Later, the plant also provided a large number of reagents for related research in my country and became an early model for the application of scientific and technological achievements.

In 1959, the synthetic insulin project was included in the national scientific research plan, and was awarded the national secret research plan code “601”. This means that the project was the number one task in the 1960s, and the leaders of the party and the country personally concerned about it. On May 5, 1960, the Institute of Organic Chemistry received a surprise mission from the Party Group of the Chinese Academy of Sciences and formally participated in the synthesis of bovine insulin.

The idea of ​​synthesizing bovine insulin was put forward during the “Great Leap Forward” period. By the time the Institute of Organic Chemistry joined, the project had basically determined the plan for “Big Corps Combat”. At that time, Wang Zhongliang, secretary of the Party Committee of the Shanghai Branch of the Chinese Academy of Sciences, was in command and commanded five research institutes including the Institute of Biochemistry, Institute of Organic Chemistry, Institute of Medicine, Institute of Cell Biology, and Institute of Physiology to conduct “big corps operations” with a total of more than 300 people. The Institute of Organic Chemistry invested a total of 140 researchers and technicians, and about two-thirds of the researchers at that time were involved in the synthesis of the insulin A chain.

But after a few months of “big corps operations”, scientists discovered that the A and B peptide chains of organically synthesized insulin involve a large number of experimental designs, and it is not simple to pile peptides. The “Big Corps” ultimately failed to synthesize the A and B peptide chains of insulin.

The leaders of the Chinese Academy of Sciences Zhang Jinfu, Du Runsheng and others accepted Wang Yinglai’s suggestion and decided to terminate the “big corps operation”, leaving the Institute of Biochemistry and Organic Chemistry to continue cooperating. Later, in 1961, Wang Zhongliang emphasized in the scientific summary of experience and lessons that we must respect the laws of science and unremittingly continue to promote the work of synthetic insulin. In the same year, Vice Premier Nie Rongzhen inspected the Institute of Biochemistry and stated that “No matter how much responsibility you do, we will bear the responsibility, and we will continue to do synthetic insulin for 100 years.”

In 2015, when the two first schools held the 50th anniversary commemorative activities for synthetic bovine insulin, Ding Kuiling, the director of the Institute of Organic Chemistry at the time, and the veteran experts who participated in this research all mentioned the “spirit of science” to an unprecedented level. the height of.

“In the beginning, everyone rushed to make synthetic insulin, and then many units did not do it. The director took a few of us and bit the bullet to pick up this project and continue to do it.” Xu Jiecheng said that the director was at that time. Wang You, director of the Institute of Organic Chemistry. Ding Kuiling, academician of the Chinese Academy of Sciences and organic chemist Dai Lixin later jointly pointed out in the article “From the synthesis of crystalline bovine insulin to the synthesis of our future” that after the end of the “Brigade Combat”, the Institute of Organic Chemistry left the products of the year for continuing Purified and analyzed, and successively sorted out 3 research papers and published them in the “Acta Chimica”, “Wang You thinks that this work is a good topic, and since the work has already started, there is no reason to give up.”

In the end, only those scholars who were willing to “sit cold on the bench” made it to the final success. Wang You led researchers Xu Jiecheng, Zhang Weijun, Chen Lingling, Qian Ruiqing, and experimental assistants Liu Yongfu and Wang Siqing to continue to promote the synthesis of insulin.

In May 1965, the insulin A chain under the responsibility of the Institute of Organic Chemistry and Peking University was synthesized and combined with the natural insulin B chain. When the A chain and the B chain synthesized by biochemistry have accumulated to a certain amount, Du Yucang, Zhang Weijun, and Shi Putao met at the Institute of Biochemistry and witnessed the use of artificially synthesized insulin A chain and B chain to do a total synthesis test, and finally The bovine insulin crystal was obtained on September 17, 1965.

Xu Jiecheng told reporters that one of the important reasons he was willing to “sit on the bench” with Wang You was that he saw the “nearly extreme” scientific spirit in Wang’s body.

Xu Jiecheng recalled that in the synthesis of chain A, in order to verify the purity of the condensation products at each step, from raw materials to each intermediate fragment, from small fragments to large fragments, Wang You required elemental analysis, chromatography, electrophoresis, optical rotation, Enzymatic hydrolysis and amino acid composition analysis, “If any of the analysis indicators cannot be reached, it must be further purified and then analyzed, and strive to pass all of them. At that time, the researchers also jokingly called this practice “passing five barriers and cutting six generals.” In the nearly 200 steps of total insulin synthesis, the impure product of any step will affect the subsequent synthesis. Only in this way can the repeatability and verifiability of scientific research be guaranteed.

When the crystallization was obtained in 1965, even though the importance of this work to obtain the priority of discovery in the world was clearly understood, in November the National Science and Technology Commission held an appraisal meeting for the synthesis of crystalline bovine insulin, led by Wang You Some organic chemists in China still believe that the evidence is not sufficient, and concluded: “It can be considered that crystalline bovine insulin has been obtained through artificial total synthesis”, and hope that “the final crystalline product of total synthesis should be further analyzed and identified in order to obtain a better For full proof”.

Since then, personnel from the Institute of Biochemistry, Institute of Organic Chemistry, and Peking University conducted electrophoresis, chromatography, immunodiffusion, antiserum neutralization test, enzymatic hydrolysis, and rabbit blood glucose testing on insulin crystals. It is no different from natural insulin. In the end, at the second meeting of the synthetic bovine insulin appraisal committee held in April 1966, scientists affirmed: “The above results fully prove that the artificially synthesized crystalline product is bovine insulin.”

Chen Changqing is one of the several persons in charge of insulin-related projects in the Institute of Biochemistry. He joined the project at the age of 24 and was only in his early 30s when he completed the project. When he recalled the project in 2015, he said that even the current academician of the Chinese Academy of Sciences Zhang Youshang was only 35 years old in the research group at that time. “Young people dare to think and dare to do, and they may even cause trouble, but the leaders are willing to take responsibility for them. Back then, it relies on youth, but now it’s even more dependent on youth.”