How to solve the scientific dilemma of "a monkey is hard to find"

  China News Weekly reporter/Li Mingzi

  Issued in the 992th issue of China News Weekly on 2021.4.19

  "There is no margin, all sold out." This is the second unfamiliar call to "buy animals" received by Zhang Wen, chairman of Jiangsu Zhaoshengyuan Biotechnology Co., Ltd. within an hour.

Beginning last year, some friends who had no business contacts also began to ask Zhang Wen to "buy monkeys" and "buy beagle dogs" for experiments, and Zhang Wen's old customers signed the next year as early as the previous year. Year's order.

  Zhao Shengli, secretary of the China Experimental Primate Breeding Development Association, also said that the value of experimental monkeys has soared from 15,000 yuan in the second half of 2016 to 62,000 yuan.

Often when discussing business the previous week, the merchants still asked on the phone, "Is it only acceptable for more than 50,000 yuan?" When I called again a week later, it had risen to more than 60,000 yuan.

"The situation is that a monkey is hard to find. More precisely, there is a price but no monkey."

  With the new crown virus raging around the world wildly for a year, the competition among countries for vaccine research and development and marketing has already begun, but what most people did not expect is that the lively and smart little monkey became the most in this battle. One of the most sought-after resources.

  The non-human primate laboratory animals represented by monkeys are the experimental objects that many scientists engaged in life science research have dreamed of.

Different scientific experiments will use different experimental animals, including the most common rodents, namely mice, cute rabbits, dogs, ferrets and piglets, as well as less cute crows, fruit flies... But if you use experiments In terms of complexity and cost, the monkey at the top of the chain of contempt.

With the help of monkeys, researchers can study basic scientific research issues in the fields of viruses, diseases, developmental biology, and brain science. The importance of experimental monkeys is especially reflected in the research and development of new drugs-any new drug must be launched on the human body. Before clinical trials, safety evaluations must be done on laboratory animals. Monkeys are one of the main forces of laboratory animals for safety evaluation.

But at the moment, a problem facing scientists all over the world is that the new crown vaccine and drug research and development tasks are urgent, but there are not many experimental monkeys that can be used immediately.

Why is "a monkey hard to find"?

  Every little monkey sold in China has a ten-character identification number (ID) assigned by the monkey farm itself, usually consisting of its English initials and numbers.

For example, the ID of Macaca Fascicularis (Macaca Fascicularis) with the largest number in the monkey farm may be MF00000011. The ID of Macaca Mulatta, the second largest in number, mostly starts with MM, and the letter of origin is often added to the ID. abbreviation.

Cynomolgus monkeys and rhesus monkeys are the two most important experimental monkeys.

  These IDs are proof of the innocence and high quality of the "commercial monkey".

It takes two generations of monkeys for a little monkey to move from the wild to a "commodity".

According to relevant regulations, each experimental monkey must have a detailed genetic file and health file, and only the second generation of artificially reared offspring and their offspring that are negative for the required pathogenic microorganisms can be used for experimental research.

  The strict requirements on experimental animals not only ensure the quality of research, but also lengthen the supply cycle of experimental monkeys.

It usually takes five years for monkeys from birth to sexual maturity, and about 5 and a half months from pregnancy to delivery, usually one litter per litter.

The baby monkey grows slowly and crawls on the mother monkey like a small pendant throughout the lactation period.

As a precious animal resource, experimental monkeys are not like industrial products that can be produced immediately after placing an order. The first batch of second-generation "commercial monkeys" can only be released for as long as 8 years, and then they have entered a relatively stable supply cycle.

"This is an objective reason for the tight supply of experimental monkeys at present and even in the next three to five years." Zhao Shengli said.

  Zhang Yuchao, secretary-general of the China Experimental Primate Breeding Development Association (hereinafter referred to as the "Association"), told China News Weekly that there are currently more than 240,000 two main types of experimental monkeys in the country. Except for young monkeys and breeding monkeys, there are actual commercial monkeys. There are about 100,000 monkeys. If the monkeys that are booked or underwritten overseas, are too young or are "menopausal" monkeys, there are only about 30,000 monkeys in the country.

  The pre-clinical trials of new drugs are the place where the amount of monkeys is the largest in the country, consuming about 25,000 monkeys each year, mainly cynomolgus monkeys.

Kang Long Chemical (Beijing) New Drug Technology Co., Ltd. senior vice president and head of the drug preclinical safety evaluation agency Wang Jufeng introduced that the international general ICH (International Coordination Conference for the Registration of Technical Requirements for Human Drugs) and other guidelines suggest that you should choose to be sensitive to drugs. Do preclinical trials with appropriate animal species.

Although it is not clearly stated that "monkeys must be used", 70 to 80% of new drugs for large molecules, that is, biological products, require monkeys for preclinical testing, and small molecule drugs, that is, chemically synthesized drugs, account for 70% of all new drugs. Monkeys will be used in the 20s and 30s.

  Under normal circumstances, preclinical trials will select two species of animals, one is rodents, that is, rats or mice, and the other is large animals, including monkeys, dogs, small pigs, etc. By comparing metabolic profiles, Preliminary experiments such as targets to determine which animal is more sensitive to drugs.

"If both the dog and the monkey meet the test conditions, they will definitely choose the dog because it is cheaper." Wang Jufeng said that although the price of the experimental beagle dog is also increasing, it is only three or four thousand yuan, which is only 1% of the price of the experimental monkey. /15.

  There is no absolute fixed number for the amount of monkeys used in a new drug in preclinical trials. On average, a 28-day toxicology experiment for repeated administration requires about 40 monkeys. If drug metabolism and other tests are to be performed, it will take another 20. Only around.

The rapid development of the biomedical industry is undoubtedly the fundamental reason for the rapid increase in the amount of experimental monkeys.

According to relevant industry report data, the number of new drug clinical trial applications and evaluations completed by the China National Drug Evaluation Center has increased from 494 in 2014 to 983 in 2019.

  According to Li Qin, chief scientist of the China Experimental Primate Breeding Development Association, the domestic sales and use of experimental monkeys has increased from 7,000 to 8,000 in 2013 to nearly 30,000 in 2019. In the past few years, it has increased by thousands. Since 2017, it has grown into tens of thousands, mainly for drug development and safety evaluation.

Basic research is another major "use scenario" of experimental monkeys, with an annual consumption of about 5,000.

  There are deeper reasons behind the difficulty of finding a monkey.

Xiong Wanhua, founder of the China Experimental Primate Breeding Development Association, said that cynomolgus monkeys account for 80%-85% of the total experimental monkey industry in China, but they are not native to China and need to be introduced from Southeast Asia.

After the outbreak of the financial crisis in 2008, the main business "export" of China's experimental monkey industry was affected. The monkeys were unsalable, but the domestic experimental monkey inventory was still growing, reaching the highest ever in 2013, with nearly 300,000 only.

It is against this background that China has suspended the import of species of monkeys.

  In 2015, China's drug regulatory reforms accelerated the approval of new drugs. The domestic research and development of new drugs showed explosive growth in the following years. The amount of monkeys used in new drug safety evaluations soared, and the stock was continuously consumed.

The use of experimental monkeys is growing rapidly, but the introduction of monkey species has been suspended for nearly ten years. Some monkey farms have seen the market benefit and even sold monkeys. In addition, lack of selection and breeding technology, domestic monkey farms generally have population aging and fertility rates. With low-level problems, production capacity began to fail to keep up with sales.

It was not until the end of 2018 that the country approved some of the import applications for monkey farms, but not long after that, the outbreak of the new crown epidemic in early 2020 made the situation worse.

  On January 26, 2020, China's State Administration for Market Regulation and other departments jointly issued the "Announcement on Prohibition of Wild Animal Trade", requiring that "locations for breeding wild animals shall be isolated and the spread and transshipment of wild animals are strictly prohibited."

China's experimental monkey import and export business has been suspended.

  "The production capacity of the two main species of experimental monkeys has begun to decline, from the total output of about 40,000 per year in the past to 30,000 to 35,000 in 2020, and the total annual export plus domestic sales is 40,000 to 50,000. In a few years, the stock will be exhausted.” Li Qin explained that some monkeys that have been booked by overseas companies in advance are also out of circulation. This is also the reason why domestic monkeys are still nervous after the suspension of import and export.

  In Zhao Shengli's impression, the "monkey shortage" has already begun as early as 2018, and the price of monkeys has become higher and higher, gradually exceeding the range of scientific research units.

Many researchers reported to the association that "monkeys cannot be bought and cannot be bought".

In this context, some large pharmaceutical companies have begun to acquire, merge, and establish their own monkey farms, buying and raising monkeys that are six months old and one year old for later use.

Expensive basic research resources

  On the slope of the southeast side of the Chenggong campus of Kunming University of Science and Technology, there is a 6-story red brick scientific research building. Enjoy delicious fruits or play with small toys for self-entertainment.

This is the laboratory of the school’s Primate Translational Medicine Research Institute, which houses more than 700 cynomolgus monkeys and 300 rhesus monkeys.

As the state key laboratory of non-human primate biomedicine jointly established by the province and ministry, it can provide primate gene editing, a variety of neurological disease models and metabolic system disease models, animal samples, primate brain MRI image data, and more Kind of integrated service.

  "If you want to carefully discuss all the research issues, monkeys are definitely not enough. In many cases, they will adjust or cut off the project according to the existing animal resources to minimize the demand for animals." Kunming University of Science and Technology Primates Niu Yuyu, deputy dean of the Institute of Translational Medicine, told China News Weekly.

In his impression, the demand for monkeys for basic research has increased significantly since 2015.

  According to reports, from January 26 to June 29, 2020, the industry association has coordinated 3551 experimental monkeys from its member units for basic research related to the new crown.

“The number of experimental monkeys in the major scientific research units is limited. Commercial monkeys were explicitly prohibited from trading during the epidemic. At that time, they had to see the approval of the Ministry of Science and Technology, which was related to the new crown research, before they could engage in monkey trading at the monkey farm." Zhang Yu, secretary-general of the Aquaculture Development Association, said.

  In May 2020, the "Rapid Development of SARS-CoV-2 Virus Inactivated Vaccine", jointly completed by a number of units led by Qin Chuan, Institute of Medical Laboratory Animals, Chinese Academy of Medical Sciences, was published in the journal Science, becoming the world's first public Reported results of animal experiments on the new crown vaccine.

The article announced that the vaccine developed by Beijing Kexing Zhongwei Biotechnology Co., Ltd. is safe and effective.

The experiment was conducted on rhesus monkeys.

  According to Niu Yuyu, since the "13th Five-Year Plan" period, in the field of biomedicine, whether it is the establishment of national scientific research projects or the research of each scientific research unit itself, primates will be involved. Everyone has realized that monkeys can be used for research. Answer more basic scientific questions that have not been found in rodents.

Internationally, the development of cutting-edge biological research such as stem cells and gene editing also requires primate experiments.

  Applying for experimental monkeys is much more complicated than applying for animals such as dogs and rodents, and must be reviewed by relevant ethics committees.

A domestic scientific researcher who is engaged in basic research said that in reality, there is indeed a phenomenon of repeated experiments with different species in order to make it easier to publish articles. This exists both at home and abroad, but it is only an isolated phenomenon.

  "It is difficult to judge whether there is a scientific research bubble in China's animal experiments based on the amount of experimental monkeys alone, but the cost of experimenting with monkeys in China is lower than in the United States. However, the number of experimental monkeys used in basic research in the United States is still far more than that in China." Basic scientific researchers told China News Weekly.

  Another major feature of monkey experiments is that they are expensive.

In 2013, Wang Xiaohua, the attending physician of the Department of Anesthesiology, Xuanwu Hospital of Capital Medical University, and others published an experimental study on the anesthesia plan for the brain and spinal cord fMRI examination of the rhesus monkey spinal cord model in the Chinese Journal of Comparative Medicine.

According to Wang Xiaohua, the key projects of the Ministry of Science and Technology that generally require monkeys are in the order of ten million yuan, and nearly one-third of them are used for the establishment of experimental monkey models.

The monkeys were taken care of by professionals in specialized institutions outside of the research period, plus labor, water, electricity, and nursing costs. At that time, the purchase and maintenance costs of each monkey had reached 50,000 to 100,000 yuan.

  "The cost is mainly in hiring professionals. The veterinarian is the spokesperson of laboratory animals and can translate animal behaviors to ensure the quality of research and animal welfare." Li Qin, for example, said that the same mouse was shaking and his ears stood up. Shaking at the same time means excitement, shaking the beard backwards is a sign of coldness, and if the beard is flat, it means pain.

In addition, different animals have different behaviors on the same issue, and these all require long-term accumulation of experience. Professional veterinarians need to conduct multiple observations every day during the experiment. Thousands of dollars.

  Monkeys, which are so expensive and difficult to apply, are still popular because they have irreplaceable value in some important basic research fields. For example, developmental biology studies how humans gradually form and differentiate from fertilized eggs into different cells, organs, and form humans. There are also studies on human degenerative diseases.

Pink-faced rhesus monkeys and smaller cynomolgus monkeys are commonly used in research on neurological diseases and reproductive development. African green monkeys have more advantages in some virus research.

  With the increase in the amount of monkeys used in preclinical trials of new drugs in China, it has gradually squeezed out the amount of monkeys originally used for basic research.

"Market demand is not necessarily representative of the country's needs. It is necessary to distinguish market demand from scientific research and national demand with monkeys to guarantee the amount of experimental monkeys for basic research." "China News Weekly", since 2015, he has submitted written suggestions to relevant departments, calling for the establishment of a national non-human primate research center, one is to use primates to study the cutting-edge basic scientific issues of life sciences, and the other is It is to establish primate animal models for complex diseases, study disease pathogenesis, and find treatment methods.

Industrial dilemma

  China's non-human primate resources are very rich. There are 4 families, 8 genera, 24 species and 45 subspecies in the country, which is about 10% of the global non-human primate species.

However, the main species of experimental monkeys were cynomolgus monkeys weighing 4 kg to 5 kg, rhesus monkeys weighing 7 kg, as well as a small number of marmosets and green monkeys.

  Why did the foreign cynomolgus monkeys replace the local rhesus monkeys and occupy the absolute dominant position in the Chinese experimental monkey market?

"This is because the monkey species used in preclinical trials abroad is a cynomolgus monkey." Li Qin explained that the cynomolgus monkey is small in size and consumes less raw materials in the safety evaluation, which can reduce costs. Therefore, follow-up studies are also inclined to Carried out with cynomolgus monkeys.

The earliest business of China's experimental monkey industry is foreign trade, so the proportion of cynomolgus monkeys has always been high, and the second-ranked rhesus monkey is only 15% to 20%.

Zhang Wen's company is the only domestic company that sells marmoset monkeys, with an annual sales volume of two to three hundred. African green monkeys, ranked fourth, are only introduced by individual domestic scientific research institutions.

  Globally, the United States is the country that uses monkeys the most in the world.

According to statistics from the U.S. Department of Agriculture, the amount of monkeys in the country hit a record high in 2017, with more than 74,000 monkeys used throughout the year, including commercial dosages such as drug safety ratings and basic experimental dosages.

The United States once mainly relied on India to supply rhesus monkeys, but since 1978, due to various reasons such as religion and animal protection, India stopped exporting to the United States.

The cynomolgus monkeys in Southeast Asia have become an alternative to imports from the United States. The Chinese monkey industry took advantage of the demographic dividend and the opportunity of reform and opening up at that time to quickly replace Southeast Asia to fill this market gap and become the world's largest non-human primate laboratory animal. Supply country.

Guangxi is one of the provinces with the largest number of experimental monkeys in China.

  According to conservative estimates, at present, there are about 48 experimental monkey farms with a certain scale in China, distributed in Guangdong, Guangxi, Yunnan, Sichuan, Beijing, Suzhou and Hubei provinces. Based on the current market conditions, a monkey farm can be sold. To more than a billion yuan.

Bei, Shanghai and Guangyin gather major universities, hospitals, and scientific research units, and are the three regions with the largest amount of experimental monkeys.

  However, northern China is not suitable for breeding cynomolgus monkeys.

Cynomolgus monkeys are also known as long-tailed macaques. In low temperature environments, the end of the tail will become necrotic due to poor blood circulation, while keeping warm and ventilating are contradictory. The indoor ammonia concentration will increase due to lack of ventilation, thus affecting Experiment on the health of monkeys.

In 2016, Zhang Wen's monkey farm moved from Fangshan, Beijing to Jiangsu due to Beijing's relief of non-capital functions.

  "The Yangtze River Delta region is the most complete experimental monkey industry chain." Zhang Wen commented that the upstream, midstream, and downstream industry chains of production, supporting, and terminal markets are complete. Pharmaceutical companies gather here and a large number of monkeys are used for safety evaluation of new drugs. In addition, from For experimental monkey feed, cages, laboratory animal room purification and sterilization equipment, to monkey exclusive welfare toy production, you can find almost any supporting enterprise in Jiangsu Province.

  After more than ten years of reverse polishing by European and American customers, the quality of Chinese experimental monkeys has been continuously improved.

"Each link has very high technical requirements to ensure that animals are treated humanely and scientifically." Zhang Wen, for example, said that his company has obtained the international laboratory animal evaluation and evaluation recommended by the US Food and Drug Administration and the European Community. Accreditation Committee (AAALAC) certification.

Take the longest distance between China and the United States as an example. They choose professional monkey cages to ensure that the monkeys have food and drink along the way, suitable space, and good ventilation. After landing, a professional third-party transportation company will be responsible for picking up the airport. The special car has air-conditioning control. Wen, the veterinarian needs to follow the car.

  A veteran in the industry with 17 years of experience pointed out that the current suspension of the export of experimental monkeys by the country is objectively forcing customers to turn to Southeast Asian origin, but has played a role in helping the development of the local experimental monkey industry.

Once relevant companies in Southeast Asia and other countries get funding and mature technology, the scale of experimental monkeys is likely to surpass China in a few years. At that time, it will be difficult for Chinese experimental monkeys to enter the international market.

What is more worrying is that the monkey farms of origin have a multi-funded background, will they restrict the introduction of monkey species in China in the future, and eventually get their necks stuck?

"This is why we are now calling for the liberalization of monkey imports." Zhang Yuchao said.

  On the other hand, the association has repeatedly proposed measures such as "exchanging populations between monkey farms" at industry conferences to avoid inbreeding monkeys, urging companies to improve monkey population selection, conservation and breeding levels.

For example, Li Qin said that although it is also impossible to introduce provenance, a monkey farm in Zhaoqing, Guangdong attaches great importance to selection and breeding. The monkey files are clearly recorded, so there is no problem of population aging.

Strategic reserve

  At the beginning of this year, Ren Jin, a representative of the Shanghai Municipal People's Congress and director of the Center for Drug Safety Evaluation of the Shanghai Institute of Materia Medica, Chinese Academy of Sciences, said in an interview with the media that at present 90% of experimental monkeys in China are controlled by private companies.

She suggested that Shanghai could start a government designated procurement plan for experimental monkeys. For example, can relevant departments go to existing experimental monkey breeding and breeding bases in Hainan, Yunnan, Guangdong, Guangxi and other places to incorporate the resource allocation of experimental monkeys into the government’s resource allocation plan. First, meet the needs of national strategic projects.

  "Our statistical data is even higher than this. 93% to 94% of experimental monkeys are raised in private, private or foreign companies." Li Qin said.

At present, the only non-human primate laboratory animal resource bank in China is the Primate Laboratory Animal Center of the Kunming Institute of Zoology, Chinese Academy of Sciences. There are a total of three to four thousand cynomolgus monkeys and rhesus monkeys, but these monkeys are not used for market circulation. The resource bank The most important meaning is to preserve the seed.

  The United States realized the important strategic position of experimental monkeys as early as 2002.

The National Institutes of Health (NIH) has funded 7 national primate research centers, with approximately 35,000 non-human primates, including 25,000 experimental monkeys, mainly for basic research.

In addition, the United States imports more than 30,000 primates, mainly cynomolgus monkeys, from China and Cambodia each year to meet industrial needs such as pre-clinical trials.

Li Qin once calculated that even without imports, the 7 primate research centers in the United States have nearly 10,000 newborn monkeys each year. At the same time, research projects funded by NIH basically select monkeys from these 7 centers, achieving a virtuous circle. , Guarantee the use of monkeys for basic research.

  Therefore, "Some time ago, foreign media reported that China stopped exporting experimental monkeys and stuck the neck of the United States. This statement is wrong." Ji Weizhi said.

  As for how China establishes a strategic monkey reserve, an expert from the association, who did not want to be named, told China News Weekly: “China must integrate its own national conditions without copying American experience.” The expert said that in the long run, it must improve The technical content of experimental animals, the development of animal model products with independent intellectual property rights, rather than purely selling animal resources.

On the other hand, we must master the right to speak in formulating technical rules.

For example, the AAALAC certification recognized by the international experimental monkey market requires 300,000 RMB for registration, and China needs to explore its own certification brand.

In addition, it is necessary to expand the strains of experimental animals, develop macaques (including rhesus monkeys) as the main experimental animals, and try to develop more strains besides cynomolgus and rhesus monkeys.

  But developing an experimental monkey model is not easy.

Zhang Wen explained that monkeys, as a relatively advanced animal, have a much more complicated genetic background than mice and other animals, and it is difficult to produce relatively stable genetic characteristics through methods such as inbreeding and closed groups.

In addition, even if a model can be built through intergenerational inheritance, it will take more than ten years or even decades to calculate the life cycle of monkeys.

  “That’s why the support of the central government is needed.” Officials from relevant departments stated that in the 2021 project application guidelines for the “Fourteenth Five-Year” National Key Research and Development Program, “Basic Scientific Research Conditions and Development of Major Scientific Instruments The layout includes research on the creation of human disease animal models, the improvement of the service quality of the National Laboratory Animal Resource Bank, and the research on key technologies for the evaluation of laboratory animal quality. It will allocate 100 million yuan to support it every year for five consecutive years.

  In January 2018, Sun Qiang's team from the Institute of Neuroscience of the Chinese Academy of Sciences successfully cultivated the world's first cloned monkey with individual cells.

Sun Qiang once said that the use of cloning technology can breed a large number of model monkeys with the same genetic background, which can not only meet the urgent needs of brain diseases and advanced cognitive function research, but also be widely used in new drug testing.

  "This is indeed a new direction, but it is still too far away from being used for security reviews." A senior industry insider said that the technology of monkey cloning is not popular yet, and the cost is much higher than the market price. The biggest problem is that it is "unavailable."

  "Leasing imports is currently the fastest solution we have seen to solve the shortage of monkeys, or partly alleviate the current tension." Li Qin said, another important thing is that the import procedures have been completed and completed in 2019. A group of companies that made payment but did not have time to ship the monkeys into the country due to the epidemic situation urgently need to be released. Otherwise, if it is delayed, the surrogate maintenance fee charged by foreign suppliers will far exceed the price of the experimental monkey itself, and this account loss will change. It became a real loss.

China News Weekly, Issue 14 of 2021

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