Medicines for export

Projects developed at Unicamp will be presented by Inova in the United States

Imagem: eduardo cesarLeaf and extract of Bidens alba: anticarcinogenic properties in testsImagem: eduardo cesar

Three projects with innovative results in areas as different as phytochemistry, new materials and biotechnology have been chosen by the Innovation Agency of the State University of Campinas (Unicamp), Inova, to be presented at the TechConnect Summit 2006. Scheduled for May 8 and 9 in Boston, in the United States, the event brings together representatives of patent offices of American institutions like the University of Boston, the University of California and the University of Minnesota, the University of Stockholm, in Sweden, and the École Polytechnique Fédérale in Lausanne, Switzerland, amongst others, as well as companies like Basf, IBM, Motorola, Ford and investors in search of good business opportunities.

“It is the first experience we have of taking part in an event of this nature to evaluate the attractiveness of Unicamp’s technologies in an international scenario”, says Professor Roberto Lotufo, Inova’s executive director. The participation takes on a special meaning, because it makes it possible for a Brazilian university to take part in the TechConnect in a pioneering way. “We are opening doors for other universities and expanding the opportunities for marketing new technologies.” In the submission process, in which proposals from all over the world compete, Inova exhibited five technologies, and had three of them approved. Quite a noteworthy number, considering that only 40% of all the innovations presented were chosen.

The criterion for selection at Unicamp, which has 425 patents deposited, took into account the most recent patents that fitted the demand from the market, such as a new phytotherapic drug extracted from Bidens alba, popularly known as common beggarticks, with promising results for some kinds of cancer and leukemias (see the article on Unicamp’s leadership of the patents ranking on page 28).

The second technology is a peptide, a molecule made up of 12 amino acids that fights the parasites that cause avian coccidiosis, a disease responsible for delayed growth of farm poultry and for bringing losses to the sector. The last patent chosen is an adhesive for metals like aluminum and steel, which dispenses with the prior treatment of the surfaces to be glued.

“Nowadays, the international companies have business executives who cover the world in search of a good technology”, says Rosana Di Giorgio, Inova’s director for Intellectual Property and Development of Partnerships. “That is why it is important to show these companies that Unicamp has good technologies, is willing to license them, and Brazil has favorable laws for this to occur.”

The study of the phytotherapic drug started in 2000, when researcher Maria Tereza Grombone Guaratini, under the Biota program, financed by FAPESP, began studying in her postdoctoral project the genetic variability of hairy beggarticks (Bidens pilosa), the most important invasive plant of soybean crops. “The study of the chemical compounds found in the plants, such as the sesquiterpenes and polyacetylenes, associated with chromosome research, made it possible to prove the existence of three species”, says Maria Tereza. Besides two species of hairy beggarticks, the researcher found that there was a third one, Bidens alba, original from Mexico and found only on the coast of São Paulo. The explanation for the restricted location of the plant in Brazilian territory is that it must have been brought by ships that docked at Santos.

In informal conversations about the plant, the researcher heard several people say that they had already used hairy beggarticks in the form of a plaster, to combat muscular pains, or even as tea. In the scientific literature she also found references to its antimicrobial, antihelminthic (against worms) and antiulcerogenic action of Bidens pilosa, in studies that looked for explanations for using this species in medical practices in Africa and in the Amazon. As Bidens is an invasive plant, it occurs in various countries of the world in agricultural areas or in places that have been environmentally modified. “The majority of the work was carried out by chemists who were speculating about the action of compounds found in the plant or by pharmacologists who were testing the extracts on animal models, without, however, verifying the real content of the extract”, says Maria Tereza. In the literature researched no reference was found to Bidens alba, which was revealed in the studies carried out by the researcher as a plant chemically different from the other two species of Bidens pilosa studied.

Maria Tereza then went to show the extract from the plant to Professor Alba Regina Monteiro Souza Brito, from the Physiology Department at Unicamp, who suggested taking the material to the Physiology Department of the Biosciences Institute of the São Paulo State University (Unesp) of Botucatu. There, a former student of Alba, Professor Clelia Akiko Hiruma Lima, of the Natural Products Laboratory, was willing to do tests with Bidens alba. Tested on mice with acute gastric ulcers, the dark green liquid worked like an excellent protector of the gastric mucus. “In the comparative tests, the extract from Bidens alba responded better than the commercial product most used for gastric ulcers”, the researcher says. The patent deposit of the extract with antiulcerogenic activities was made by Inova in 2004. Some companies have already been in contact with the agency, interested in carrying on with the work and in developing products.

Imagem: eduardo cesarAdhesives glues metals without prior preparation and is water- and heat-resistantImagem: eduardo cesar

The researches with Bidens alba have not stopped at this point. To find out whether the extract also showed an anticarcinogenic effect, the researcher took the liquid to be evaluated at Unicamp’s Integrated Center for Childhood Onco-Hematological Researches (Cipoi), a basic and applied research laboratory intended for studying children suffering from acute leukemias. As the extract is divided into 73 fractions and the chemical composition of each one of them is not known with precision, three were chosen to be studied. Two of them showed very promising results.

The work of chemically separating each fraction was done by researcher Carmen Lúcia Queiroga, from the Pluridisciplinary Center for Chemical, Biological and Agricultural researches (CPQBA), also at Unicamp. When they received the extract, researchers Alexandre Eduardo Nowill and Gilberto Carlos Franchi Junior, from the Cipoi, a physician and a pharmacist who dedicate themselves to researches into leukemia, limited themselves to testing the new product, without knowing what it was.

After the initial results, when they saw that the extract really did work and was promising, the two researchers communicated to Maria Tereza that they were going to carry on with the research. The tests included various lineages of cancer, including adenocarcinoma of the prostate, one of the main types of this disease that affects men. “About 50% of men are going to have prostate problems, and the medicines that work for this kind of cancer are rare”, Franchi Junior says. “So if there is one that works, at least in vitro, it is already a good result.”

The tests on animals will be started next month, a stage that consists of inducing human prostate cancer with the same cells used in vitro in animals specially developed for this purpose. After 18 days from inoculation, that is to say, soon after cancer is diagnosed by palpation, the animals will begin to be treated with the drug. At the end of the treatment, the tumors will be examined by an accredited pathologist. The team is enthusiastic over the fact that the animals that received the extract in the research into the cicatrisation of ulcers did not shown intoxication in the first 24 hours of treatment.

Besides evaluating the effectiveness of the extract for fighting prostate cancer in vitro, studies were also made for breast and ovarian cancer and for four types of leukemia. The comparison was always done with a chemotherapic drug existing in the market. “The extract of Bidens alba worked for all the adenocarcinomas studied and for the leukemias”, Franchi Junior says. The research is currently at the reproducibility stage, with new in vitro tests. The multidisciplinary team made up of a botanist, a pharmacist, a physician and a chemist depends on partnership with private enterprise, which they hope to achieve as soon as a technology transfer contract is signed.

The patent deposit for the new Bidens alba based phytotherapic drug with antineoplastic activity was made at the end of 2005. And it is this capacity of the extract for inhibiting the growth of the carcinogenic cells that will be presented at the TechConnect on May 8.

Slow growth
The antimicrobial peptide project is the result of a research carried out at the Molecular Biology and Genetic Engineering Center (CBMEG) and in the Parasitology Department of Unicamp’s Biology Institute. The peptide, which is a fragment of a protein, is a sure-fire combination of 12 amino acids baptized as PW2 (see Pesquisa FAPESP No. 78). It is capable of destroying the protozoon of the Eimeria genus, a unicellular organism and a causal agent of coccidiosis, a disease that acts on the digestive processes of chickens and prevents a greater absorption of nutrients by the animal. The project began from the doctoral thesis of researcher Arnaldo da Silva Júnior from the CBMEG, supervised by Professor Adilson Leite, who died at the beginning of 2003, and which was able to count on the participation of professors Urara Kawazoe and Paulo Arruda.

The disease, one of the main ones to affect birds, has direct consequences for the productivity of the poultry industry. “When the birds are infected by the parasite, the rate of growth is far slower than in a healthy bird, which means that they are discarded as waste”, says Urara, for the Biology Institute at Unicamp and co-supervisor of Silva Júnior, who works today in a private sector company in Indaiatuba, a town close to Campinas. The protozoon causes losses in the absorption of important nutrients for the normal growth of the birds, which ought to reach the ideal weight of 2 kilos with about 40 days at the time of slaughter.

The attempt to control avian coccidiosis in the commercial poultry farms includes the use of vaccines and, on a smaller scale, the administration of anticoccidial preventive medicines. The effect of these medicines has been inefficient, due to the presence of resistant strains of the parasite. Furthermore, the use of these medicines in birds intended for foreign trade is not permitted, according to the legislations of the countries importing these birds.

The protection offered by the PW2 peptide is related to the capacity of this protein structure to damage the protozoon’s protective membrane before it manages to invade the animal’s intestinal cells and  consequently to prevent the development of the disease. When there is no barrier capable of fighting coccidiosis, the parasite takes an average of four or five days to complete its life cycle, which begins with the elimination of the immature oocysts, a way of resisting parasites, together with the feces of the chicken. The oocysts mature in the environment, creating in their insides the infectious forms of the parasite, called sporozoites.

When the birds scratch around in the ground, they acquire the parasite by ingesting mature oocysts, wrapped in two resistant outer membranes that are impermeable to liquids, as well as another double membrane that encloses the sporozoites, the initial form of the parasite. Passing through the chicken’s digestive apparatus, known as the gizzard, the oocyst is ground up, bursting the outside membranes. From then onwards, by the action of the digestive enzymes, the sporozoites, the first asexual form, stick to and penetrate into a cell of the intestinal mucus and are covered by a membrane of the host itself, forming a wrapping around it. “In this way, the parasite stays well protected, and the host itself does not recognize it as a foreign body”, Urara explains. “It is an escape mechanism of the parasite.”

Getting peptides with the right “fit” for sticking to the protozoon’s membrane took four years. The researchers selected various series of amino acids from a great library of peptides for coming into contact with the parasite. When the 12 amino acids of the peptide were tested in vitro, they worked as if they were an antibody, with power to act as an antimicrobial agent.

Besides destroying the initial stage of the parasite, the peptide does not leave chemical residues in the meat, because it is absorbed by the organism as a protein. This benefic factor is important in the international market. The importers, mainly European and Asian, raise trade barriers for poultry with residue of substances used for treating diseases. “Our intention is to supply the peptide together with the feed given to the birds”, Urara says.

But, to reach this point, another two stages still need to be covered. One of them is to do the final tests with chickens, to start with at the university’s vivarium and afterwards on a poultry farm, because up until now the experiments have been done in vitro. And to establish the ideal concentration for getting the best result. To do so, the peptide has to be synthesized on a large scale, a stage that needs a partnership with companies, for the high cost it entails. The patent that ensures the rights over the use of the peptide for fighting coccidiosis, its variables and the method used for identifying it has been deposited in Brazil, the United States and Europe, with the help of FAPESP’s Patent Licensing Nucleus (Nuplitec).

Imagem: eduardo cesarBirds from the poultry farm are infected by the Eimeria protozoonImagem: eduardo cesar

Structural differences
Also to be presented in Boston is a polymeric adhesive formed by a nanostructured acrylic polymer that sticks hard to aluminum and steel and that therefore has a good potential for replacing screws, nuts and rivets in metallic surfaces. The applications include the metalworking, automobile, aeronautical, civil construction and furniture industries.

To arrive at the new adhesive, researchers from Unicamp’s Chemistry Institute, coordinated by Professor Fernando Galembeck, dedicated themselves to systematically studying a family of polymers that, although similar, show small structural differences caused by the change of one of their components, called a tensoactive, which is used in the manufacturing process. “The tensoactive changes the structure of the polymer on a nanometric scale”, Galembeck says. The change in the nanostructure changes the interactions between the polymer and the metal, as well as the mechanical properties of the polymer itself, making the adhesive joint more resistant to mechanical demands.

In one of the tests carried out in the characterization process, the polymers were passed though an extrusion machine, much used for processing plastics. When two of the resins from the family studied by the researchers were used, the technician responsible had difficulties in cleaning the equipment’s screw. “We went to see what was happening and we noticed that there was a very strong adhesion of the resin to the steel of the screw”, Galembeck says. “It was a problem that in the end proved to be a solution.” Although it has in its composition many substances present in other products of the kind, the new adhesive has as a differential the advantage of dispensing with the prior preparation of the metal surfaces that will be glued.

Gluing metals like aluminum and steel usually requires, besides cleaning, various treatments that create a chemical structure on the surface that favors the fixing of the adhesive, but this may impair the stability of the object, particularly when there is a lot of humidity in the air or in extremely hot environments. In the case of the new adhesive, stability is not the only advantage. It also shows good resistance to water. In the laboratory of the Chemistry Institute, there are several parts glued with the polymer that have been immersed in water for over two years. “The metal is very stained, but the part covered by the adhesive is perfect”, Galembeck says. The patent that deals with the process for manufacturing the product was deposited at the beginning of 2005.

Tests in the laboratory have now been done to prove the adhesive’s resistance to stresses  from flexure and torsion, both when dry and in a humid environment. “Now we have to do field testing on a large scale to evaluate the adequacy of the product to the demands of the market”, Galembeck says. Although they are used in small quantities, adhesives have a turnover in the world market estimated at US$ 18 billion, half of which is in the United States. And it is in these markets that Inova is interested in trading the patent.

International business
The presentation at the TechConnect is one more stage in the search for international partnerships being undertaken by the Innovation Agency. Last year, representatives of Inova were at the Licensing Executives Society (LES), held in Phoenix, in the United States, a business conference that deals with intellectual property and technology transfer and attracted the participation of over 20 companies of the size of IBM, HP, Roche and the like. The LES is organized by the United States and Canada and is held in various different countries. This year, the event was held from April 9 to 12 in Seoul, in Korea, and once again the Innovation Agency was present. “It was an opportunity for beginning to have contact with Asia”, Rosana Di Giorgio.

At the event held in Phoenix, Intellect Ventures, a small company located in the United States that is looking for new technologies in accordance with the demand and has customers like Microsoft and Intel, turned to Inova to propose a partnership in the intermediation of patents in the information technology area. Remuneration only takes place when the deal is closed. For the time being, they have to await the result of the first notice published in the Official Gazettes of the State and Federal Governments, in March, with the new technologies developed at Unicamp. The notice is a requirement of the Law on Innovation, regulated in October 2005, and it precedes the licensing of the technologies developed by public bodies. *The technologies that are not licensed following the notice will be offered by Intellect Ventures to the American market”, Rosana says.

The Project
Patent request for a new method for selecting antimicrobial peptides and the anticoccidial peptide PW2
Intellectual Property Support Program (Papi)
Paulo Arruda – Unicamp
R$ 26,875.29 and US$ 48,320.07