Bioengineered liver hits barriers

A blow to the liver

Lyudmila Kolbina, "Expert Ural" No. 11-2013

Biomedical Technologies LLC (Miass), engaged in the development and creation of a multifunctional device with biomaterials for blood purification, became the sixth South Ural company to receive the status of a resident of the Skolkovo Innovation Center. The company, based on the Miass plant of medical Equipment, has been creating a device for extracorporeal blood purification "Bio-artificial liver" for ten years.

Vyacheslav Ryabinin, professor of the Biochemistry Department of the Chelyabinsk State Medical Academy, tells E-U why, despite loud statements from above, breakthrough biotech projects go in a vicious circle.

Why is the device outdated– Vyacheslav Evgenievich, the Ministry of Defense of the Russian Federation on the public procurement portal recently announced a competition for the creation of a bioengineered liver.

An artificial organ using the patient's own stem cells is needed in hospitals for transplantation. The winner of the tender will be named on March 20, the work must be completed by November 25, 2016. The price of the issue is 518.5 million budget rubles. Will you participate?

– Our research has a lot in common with this project. But it looks very much like an adventure. According to the tender documentation, in three years the army should receive a "bioengineered liver", the production period of which for each patient is two months. To conduct R&D, animal research and three stages of clinical trials in the absence of experience in the development and creation of such systems, a regulatory framework for biomedical technologies in the Russian Federation is out of the realm of fiction, even for such money. There is no doubt that the contest is tailored to an already well–known performer. Therefore, competition from our side is meaningless.

– What about your project?

– As residents, we are applying for a grant from the Skolkovo Foundation of 30 million rubles. To receive it, a quarter of this amount must be paid by the applicants, that is, a co-investor is needed. It is the Miass Medical Equipment Plant: General Director Vladimir Suprun is ready to invest more than 7 million rubles in the continuation of the project – the creation and manufacture of a new device for extracorporeal blood purification.

– Why are we talking about a new device already?

– Small amounts of funding did not allow us to implement the project quickly. Money had to be found wherever possible. The first device appeared in 2002 with the participation of the medical equipment plant with the support of the RFBR grant. Then, with grants from the Foundation for the Promotion of Small Forms of Enterprises in the Scientific and Technical Field (Bortnik Foundation), RGNF and RFBR, we conducted R&D and two phases of clinical trials in 2009 and 2010 on the basis of the South Ural Scientific Center of the Russian Academy of Medical Sciences and the Chelyabinsk Regional Clinical Hospital. We treated 20 people with liver failure and got good results. It remained to pass the third stage – clinical trials at the Federal Scientific Center of Transplantology and Artificial Organs named after Academician V.I. Shumakov in Moscow.

But the world has gone far ahead in ten years. At the beginning of 2011 it became clear: the device "Bio-artificial liver" is outdated, a new instrument base is needed. If you conduct the third stage of clinical trials on it and get a certificate, you will have to produce and sell it in this form, but you cannot do this without modernization. The management of the plant, taking into account what is on the world market today, said: we will not produce it, because no one will buy it. Therefore, we have made a very painful decision – not to conduct third clinical trials yet, to make a new device, more modern in terms of electronics, actuators, procedures for the treatment of patients. Innovative biomedical technologies, including cellular ones, will become an integral part of the medical complex.

In addition, there is no technical possibility to produce an outdated device: it was created using components manufactured at enterprises using Soviet technologies. The factory can't buy anything like that in the country now. And it is impossible to buy components for a more advanced extracorporeal blood purification device abroad – they do not sell. Miass is currently developing and manufacturing prototypes of everything necessary. This is another of the most powerful obstacles and complications of our task. We create everything in a single copy in order to understand how technologically it will be, acceptable in quality, and so on.

– What is the modernization?

– We are going to significantly expand the functions of the former "Bio-artificial Liver", to make a multifunctional blood purification device in which extracorporeal (outside the body) cellular and bioengineering technologies would be used as therapeutic agents. There are still no such devices on the market. The most advanced American companies working on the creation of analogues, conduct only the first phases of clinical trials. So we still have a chance to be here first, to conquer the market.

Part of the functions of the device we are developing will be like the German "Artificial Liver" device. It is very expensive: a session costs 4 thousand euros, and it takes four to five sessions to detoxify. Because of this, it is almost impossible to use it in a regular clinic. Our price should be an order of magnitude lower. We also propose to increase the clinical capabilities of the device by performing the functions of hemodialysis, hemofiltration and plasmapheresis. And the main feature is that we are going to connect cellular technologies and tissue engineering to this device. This is basically absent in the market of blood purification devices. In Russia, no one is doing this except us now, everyone has the same problems – money and personnel.

The essence of the innovation is as follows: we isolate individual isolated liver cells – hepatocytes, grow them in special conditions. In the apparatus, the patient's plasma or blood contacts these cells. With conventional hemodialysis, blood passes through a hemodialysis cartridge, inside its hollow fibers it is washed with dialysis fluid and toxic compounds that accumulate in renal, hepatic insufficiency are washed out, getting into the compartment with water. Instead of a compartment with water, we connect different solutions: with albumin, which absorbs toxic substances, with cellular systems that take over the function of detoxification and secrete biologically active molecules that contribute to the normalization of human metabolism. This is fundamentally important – not only cleansing, but also normalization.

– Is there a risk that the situation will repeat and the new device will become obsolete?

– I wouldn't like that. But there is a risk. If Skolkovo gives a grant, it's fine, if it doesn't, nothing will happen. We also need more powerful, at times, support from the state. Because without it, even progressive-minded businessmen and plant directors will not invest money in scientific developments of biotech, which it is unclear when they will give any benefit. It is impossible even to compare how our team works and, for example, the Fresenius Holding (Germany), which has 150 branches all over the world, its own research institutes. The medical equipment plant in Miass finances the project as far as possible, but it has its own production and its own tasks. Therefore, external financing is needed.

There are no stairs– The specifics of biotech is that it is very expensive?

– The program "Medicine-2025" involves the biologization of medicine through the introduction of new technologies. Here the basis is fundamental research, but they are all, of course, costly. Biomedical Technologies LLC has signed an agreement with South Ural Medical University on scientific cooperation, joint research: the university provided a room for the creation of a special laboratory. Unfortunately, science is not funded through an educational institution. The only possibility is grants.

– What exactly requires money?

– One experiment on isolation of isolated hepatocyte cells from rat liver and creation of conditions for their growth requires 20 thousand rubles. A huge number of experiments in the course of the study should be carried out in specially prepared so–called "clean rooms", and the level of laboratory and industrial practice should comply with the international standard - GLP and GMP, respectively. A cell freezing machine costs a million rubles, a refrigeration system with liquid nitrogen or special refrigerants costs another million. Without these devices, you will not be able to work with cells. Creation of the entire laboratory with a minimum set of equipment – 10 million rubles.

Without a set of special equipment, reagents, nutrient media, it is impossible to carry out the planned program, for example, to obtain liver cells from embryonic, in particular for detoxification systems. Bioengineering research is of particular interest. Here is an example: a special solution is passed through the cadaveric liver of a person, all the cells are washed out, a connective-woven frame remains, into which the stem cells of a patient with liver failure are launched (from the bone marrow). Under certain conditions, stem cells can turn into hepatocytes, the process of formation of the circulatory and biliary systems begins, the liver is formed for this person, but on the basis of a different structure. This is a fantasy that is becoming a reality, such research is being carried out in the world. And we are interested in this.

Thus, the creation and production of our apparatus with the appropriate biomaterials is associated with the solution of a whole range of biotechnological tasks that require serious funding. From a business point of view, these are very risky venture projects. It is extremely difficult to get long money for them in Russia.

– Have you tried?

– Many times. We applied to the Russian Venture Company, the Union of Business Angels of Russia and many others. When you say that something will be produced in 5-10 years, interest immediately disappears, money is offered only for two or three years and returned with interest. But biotech is not an area where it can be done quickly. This is the difference between the Russian venture business and the Western one: they can finance promising projects for 10-15 years.

– So biotech doesn't have a venture ladder for various stages of the project?

– The only exception is that they are given for medicines: the pharmaceutical business is a completely different story, a different market, this is not regenerative medicine for you. The grant from the Skolkovo Foundation, which we are currently trying to obtain, will be directed to R&D, the creation of a prototype of the device. And for clinical trials (10-15 million rubles), the creation of production and its scaling – get it yourself.

The staff of our department received financial support from the grant of the Federal Target Program of the Ministry of Education and Science of the Russian Federation and the opportunity to purchase consumables. In Miass, no one officially works at Biomedical Technologies LLC, although they are actually engaged in this topic. We will receive a grant – the plant will allocate not just money, but also designers, technologists to create a prototype, then the work will go much more actively.

Stalemate– How will the new device help a person with a sick liver?

– He has three main directions. The device is intended primarily for the treatment of patients with acute liver failure resulting from poisoning, serious diseases, injuries. Our dream is to eventually create special centers for the treatment of tens of thousands of patients. It will also be possible to conduct hemodialysis sessions on the device, as it is now on the "Artificial Kidney" device. And the third possibility of clinical use is detoxification operations that are not related to liver diseases, but where blood purification is also required and cellular, tissue engineering systems that we are currently developing can be used.

– What is the know-how?

– Firstly, only we use cytosol of crushed and specially treated pig liver tissue for detoxification, which is injected instead of dialysis solution. Clinical and experimental studies have shown the effectiveness of this biomaterial in the treatment of liver failure. Secondly, the combination of more than three functions in one device ("Bio-artificial liver", "Artificial kidney", plasmapheresis) is fundamentally new, and it is very difficult to do this. Thirdly, the know–how and what we do in relation to tissue engineering and cellular systems.

As Peter Kapitsa said, "lack of funds sharpens the mind." In the course of research, we are constantly coming up with something new. And we do not have the colossal capabilities of our Western colleagues. And, getting out of the situation, we involuntarily begin to create some new systems. Here is an example: we cannot grow cells in three-dimensional space with the help of special microfactories, it costs up to 200 thousand dollars. We sit and think about how we can do it all by ourselves, so that it would be cheap. As a result, we learned how to get collagen... from the tails of rats, make matrices for cells, found reagents for encapsulating cells, and so on. This is how many discoveries are made in Russia. Unfortunately, these passing things take a lot of precious time to the detriment of the main project, and our inventions, devices and know-how have time to grow old.

Three companies abroad are now reaching the level of clinical trials with cellular systems. But one uses hepatomas – liver tumor cells - as a source of cells. This makes us wary. The second uses embryonic stem cells, in which there are molecules whose action after they enter the patient's body is difficult to predict, this is also not very good.

Our source of cells, again based on our Russian characteristics, is most likely to be pig liver cells. It is clear that protection from porcine viruses and so on will be needed here, but this is the most real. Because the best source, it would seem, is human liver cells, but for this you need to take a cadaveric liver, and there are known problems with the possibility of taking this kind of tissue. Most importantly, the lack of adequate laws on transplantation and biomedical technologies significantly hinders the development of cellular technologies, tissue and bioengineering, regenerative medicine. The draft federal law "On Biomedical cell technologies" has been in the State Duma without consideration for several years.

The situation is stalemate. When I spoke at the Institute of Transplantology at a conference, the only question was asked: how did you manage to get permission for clinical trials. This is one of the main problems in the promotion of new medical technologies.

– And how did you circumvent the law?

– We didn't go around. We received permission for clinical trials in 2007. Until last year, it was difficult, but possible, to register a new biomedical technology with the Federal Service for Supervision in the Field of Healthcare, and after that it was difficult to obtain permission for clinical trials. And since last year, you cannot register a new technology, because there is no official structure that registers it. If a doctor uses new technologies without registration, it's on his conscience…

– How do colleagues act abroad, are there such obstacles there?

– In almost all countries there is a clear, transparent regulatory framework for the introduction of new medical technologies. There are countries where it is easier to conduct clinical trials. For example, American companies that develop devices such as "Bio-artificial liver" create branches in Singapore or China and conduct clinical trials there.

– Can't you go to China?

– There is no guarantee that not only your device is there, but you yourself will not be copied and quickly made, and then they will sell. For us, the global task is to develop technologies that guarantee the stability and repeatability of the quality of obtaining biological products. This is fundamentally important. Only then will it be possible to enter the market, into broad practice.

– Is the know-how patented?

– We have already received several patents and are processing the next ones, but in order to reach a good international level, it is necessary to equip the laboratory and production according to the criteria of GLP and GMP. Even the Skolkovo millions that have not yet been received are a drop in the ocean, at least ten times more is needed. And only in this case the movement will be obvious, we will have time to do both science and conduct clinical trials. Because if you go the way you are now again, I'm afraid it will be the same. The next prototype will become obsolete, and the West will be ahead of it.

– And why wouldn't the Miass plant tear off more money from itself, create a unique product, bring it to world markets and earn, quickly return the funds?

– He can't risk significant sums on science. The company must develop, pay salaries, fight with competitors, improve the quality of basic products. The medical equipment plant is not a venture company or a bank, it is an operating enterprise, it is engaged in the development, production, installation of "clean rooms" for medicine and pharmaceuticals. Now we have a convergence of interests with the plant: if we manage to get a grant from Skolkovo, we will step forward, nothing will succeed within a year – the topic will simply have to be closed.

What feeds optimism– What is the demand for the product that your project offers?

– Western analysts believe that the demand for Artificial Liver devices is in second place after the demand for the Artificial Heart device, the demand of the potential market for our device is $ 4 billion. Every year in Russia , 25 – 30 thousand patients with hepatic insufficiency are fatal in 80-90% of cases.

Despite the fact that dialysis centers are being created in every million-plus city, we have not succeeded in reaching patients with renal insufficiency. And if we consider that all medical equipment of this kind is 100% supplied from abroad, then the problem of import dependence becomes obvious. As for liver failure, five or six devices of the "Artificial Liver" type are working in Russia, the rest are standing, since they are not provided with consumables from the FOMS. This once again indicates the timeliness and necessity of creating domestic equipment of this type.

– Is there any way to speed up this process?

– So that we can start implementing patents and know-how faster? It's a question of money again. Yes, you can write out several specialists from Western Europe for a large price, who will do what we have been doing for years, with the availability of equipment and consumables, in two or three months. They prepare "sandwiches" from hepatocytes in vitro: a layer of liver cells, then collagen, then hepatocytes again, etc. And these cells begin to live, form bile capillaries, blood capillaries. Beautiful! We are slowly and hard going to it ourselves. Successful executives of American biotech firms say: "You should not hire middle–class people - this is the way to nowhere. Take only the best!". And we have to spend a lot of effort and money on employee training. But their best part leaves the country when there is an opportunity to improve their social and financial status. And we are starting a new cycle of teaching others.

– How are patients with liver failure treated today?

– Patients with acute liver failure are treated in intensive care units (mortality 70-80%), as for chronic liver failure, there is currently no effective treatment for this condition. That is why it is urgent to create special departments and centers, which I have already mentioned.

– Are there more problems with the liver in the population?

– Every year 3-4 million people die in the world from the effects of hepatitis and cirrhosis, in Russia 200 thousand people are admitted to hospitals annually with a diagnosis of liver failure.

– Taking into account the experience of implementing your project, do you continue to tell students about biotechnology with optimism?

– They say that a person's character is his destiny, so my optimism can only be killed together with me. When I give lectures on cellular technologies to students, teachers, doctors, it supports me, because these technologies are the future, we need to do this. And I want to convert these people.

Portal "Eternal youth" http://vechnayamolodost.ru18.03.2013