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Development story

Development story

Birth of the EX-polyphosphate®
The birth of the EX-polyphosphate®(EX-polyP®) dates back to the university laboratory over a quarter of a century ago. Please take a look at the trajectory of achieving productization after conducting a vast amount of research activities.

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Studies that no one has accomplished in the world

Challenge of Professor Arthur Kornberg, an authority on DNA research

Toshikazu Shiba, the president of RegeneTiss Inc., obtained his Ph.D. from the Graduate School of Medicine in Osaka University. He then later became a scientist conducting research and education on molecular biology at Hokkaido University. In 1993, T. Shiba worked as a postdoctoral fellow under Professor Author Kornberg in the Department of Biochemistry at the Stanford University School of Medicine, USA, where Professor Kornberg is the world-renowned authority and Nobel laureate in research on DNA replication. They touched upon “polyphosphate” as a new research topic. At that time, there were only a few scholars in the world to research on “polyphosphate”. Although the existence of polyphosphate in Escherichia coli was previously confirmed, its function and its existence in the human body were still unknown. Nevertheless, T. Shiba was highly attracted to investigating this unknown substance that almost no one could elucidate. He was urged to confirm the presence of polyphosphate in the human body, clarify its roles and use it to help people around the world.

Unknown methodology for elucidation of “mysterious substance”

Despite the fact that “polyphosphate” has a simple molecular structure, it does not mean that obtaining analytical results is as simple as inserting it into analytical instruments. At that time, measuring method for polyphosphate was not established, and previous research necessary for analysis in the literature was insufficient. As it was difficult to detect polyphosphate, so it had taken several years to do to establish the most basic detection method, which involved chemically changing polyphosphate into another substance and then use color or light detecting methods. Therefore, it was a challenge to devise detection methods. Confirming the presence of polyphosphate in the human body has become a milestone for us to start our research on polyphosphate in vivo in higher animals including human beings.

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From regenerative medicine research to starting a venture business

Polyphosphate function varies with chain length

A study to elucidate the function of polyphosphate in the human body began. In the midst of fundamental research after T. Shiba returned to Hokkaido University, he revealed the potential of polyphosphate to enhance cell function by binding to “cell growth factor”. This was a landmark discovery that could be used in the field of “regenerative medicine”. In 2004, T. Shiba started entrepreneurs from Hokkaido University and started collaborations with other universities for joint development of drug discovery. In fact, polyphosphate has been used in the industry, mainly as food additives and incombustible materials. However, it was confirmed by research that the polyphosphate used in industrial products differed from that found in the human body. Polyphosphate is formed by linking repeated units of phosphate molecules into chains, and the “chain length” changes depending on the number of phosphate units connected in the chain. We discovered a new phenomenon that the chain length decides on the physiological function of polyphosphate. Although polyphosphate for industrial use was mainly "short" chain, polyphosphate found inside the human body was relatively "long". In other words, in order to produce polyphosphate useful for the human body such as that in the medical field, it is necessary to make "long" polyphosphate with a precisely known chain length.

Research on size fractionation of polyphosphate depending on its chain length.

We began trial and error on etlicient size fractionation of polyphosphate. In order to disseminate polyphosphate as a product, we must satisfy the conditions for mass production with low cost. After intensive investigation, we invented an unexpected method of using ethanol to separate long and short chain polyphosphates. It is a surprisingly simple manufacturing method; so simple that no one has ever thought of. This is how, “EX-polyphosphate® (EX-polyP®)” with adjusted chain length was created.

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Challenging whitening products with accumulated experience from regeneration medicine research

Challenge of periodontal disease with EX-polyP®

The joint research with universities on drug discovery has expanded its application from bone regeneration to periodontal tissue regeneration. We were devoted to research, driven by the thought that as periodontal disease is a long-standing disease with no available effective therapeutic drugs, and that if we could develop medicines that can cure gum disease by directly applying on the gums and periodontal pockets, then a meaningful contribution to the society can be made. Although various trials have progressed to the point where significant effects can be confirmed, a huge amount of expenses and time are still required to obtain approval as a pharmaceutical product. In the meantime, we wondered how we can make the research results we have accumulated so far useful in the world.

Effective polyphosphate chain length for teeth whitening

Our first focus was on drugs as quasi-drugs. If we can apply EX-polyP® to oral care products of quasi-drugs, more people can use it immediately. Hence, the regenerative medicine research accumulated so far can be realized more closely.

As we move on, we discovered that polyphosphate of a specific chain length has potent whitening effect on the surface of teeth. Since polyphosphate has the function to combine with apatite on the teeth surface, this coating effect is likely to reduce the deposition of stain and plaque. As a countermeasure to periodontal disease prevention and teeth whitening, we thought of producing a product that would satisfy many people.

There have been some manufacturers that use polyphosphate as an active ingredient in their dentifrice products. However, the beneficial effects provided by the polyphosphate in these products are limited without precise optimization of the chain length, knowing that the polyphosphate function can be altered according to “changes in chain length”. RegeneTiss, being the pioneer of polyphosphate research, is the only one that possesses different technologies to control the chain length of polyphosphate. This is how, a completely new "whitening product" was produced. Basic research on EX-polyP® for many years has finally succeeded in the form of "products".

Since we started selling our products on TV channels in 2007, we have been gradually receiving comments from many people, and we are pleased to hear that our products are being regularly patronized.

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Seeking new possibilities with researchers from around the world

EX-polyP® research begins to accelerate in the world

Polyphosphate research that Professor Arthur Kornberg pursued at the end of his research career has succeeded his will and is now fruitful. However, the possibility of polyphosphate is still immeasurable.

In recent years, international research conferences on polyphosphate research were finally established, allowing scholars from around the world to advance their research activities.

As the only company that manufactures "EX-polyP®", RegeneTiss offers materials free of charge to universities and research institutions around the world that are conducting research in order to support further investigation and discoveries on the functions of EX-polyP®.

As a leading company of EX-polyP®

EX-polyP® is a unique substance whose function varies with chain length. By precisely controlling its chain length and applying its functions for appropriate applications, we are convinced that we can produce products that will make a difference in areas such as oral care, hair care and skin care. In addition, the application of polyphosphate as a supplement may have an effect on health promotion.

In the future, we plan to use EX-polyP® to do research and development to expand the field of self-medication. As a leading company of EX-polyP®, RegeneTiss aims to open up the field of health science and contribute to the maintenance of health of people around the world.

Published journal articles and books on EX-polyP®

Articles

  1. WD. Lee, R. Gawri, T. Shiba, AR. Ji, WL. Stanford, RA. Kandel, Simple silica column-based method to quantify inorganic polyphosphates in cartilage and other tissues., Cartilage, Mar. 1 (2017).
  2. H. Ogawa, C. Kurokawa, M. Hoshino, K. Tamaoka, T. Toko, T. Shiba, A. Manabe, Bleaching agent containing polyphosphate –Influence on bleaching effect, accumulation of stain, and prevention of decalcification-., Jpn. J Conserv. Dent., 59, 197-205 (2016).
  3. T. Shiba, Inorganic polyphosphate and its chain length dependency in periodontal tissue regeneration including bone remodeling and teeth whitening., in Chapter 10 of Inorganic Polyphosphates in Eukaryotic Cells., T. Kulakovskaya, E. Pavlov, EN. Dedkova Eds. (2016).
  4. LC. Wijeyewickrema, E. Lameignere, RC. Duncan, T. Shiba, RJ. Travers, PR. Kapopara, VL. Stephanie, H. Kim, JH. Morrissey, RN. Pike, EM. Conway, Polyphosphate is a novel cofactor for regulation of complement by a serpin, C1 inhibitor., Blood, 128, 1766-1776 (2016).
  5. PR. Angelova, BK. Agrawalla, PA. Elustondo, J. Gordon, T. Shiba, AY. Abramov, YT. Chang, EV. Pavlov, In situ investigation of mammalian inorganic polyphosphate localization using novel selective fluorescent probes JC-D7 and JC-D8., ACS Chem. Biol., 9, 2101-2110 (2014).
  6. K. Doi, T. Kubo, R. Takeshita, S. Kajihara, S. Kato, Y. Kawazoe, T. Shiba, Y. Akagawa, Inorganic polyphosphate adsorbed onto hydroxyapatite for guided bone regeneration: An animal study., Dent. Mater. J., 33, 179-186 (2014).
  7. M. Koyasu, T. Shiba, Y. Kawazoe, A, Manabe, T. Miyazaki, Ultraphosphate, a potent stain control agent that is effective for both stain removal and prevention of stain deposition., Dent. Mater. J., 33, 1-9 (2014).
  8. K. Tsutsumi, N. Saito, Y. Kawazoe, H-K. Ooi, T. Shiba, Morphogenetic study on the maturation of osteoblastic cell as induced by inorganic polyphosphate., PLOS ONE, 9, issue 2, e86834 (2014).
  9. K. Harada, H. Itoh, Y. Kawazoe, S. Miyazaki, K. Doi, T. Kubo, Y. Akagawa, T. Shiba, Polyphosphate-mediated inhibition of tartrate-resistant acid phosphatase and suppression of bone resorption of osteoclasts., PLOS ONE, 8, issue 11, e78612 (2013).
  10. K. Harada, T. Shiba, K. Doi, K. Morita, T. Kubo, Y. Makihara, A. Piattelli, Y. Akagawa, Inorganic polyphosphate suppreses lipopolysaccharide-induced inducible nitric oxide synthase (iNOS) expression in macrophages., PLOS ONE, 8, issue 9, e74650 (2013).
  11. N. Sun, H. Zou, L. Yang, K. Morita, P. Gong, T. Shiba, Y. Akagawa, Q. Yuan, Inorganic polyphosphates stimulate FGF23 expression through the FGFR pathway., Biochem. Biophys. Res. Com., 428, 298-302 (2012).
  12. D. Morimoto, T. Tomita, S. Kuroda, C. Higuchi, S. Kato, T. Shiba, H. Nakagami, R. Morishita, H. Yoshikawa, Inorganic polyphosphate differentiates human mesenchymal stem cells into osteoblastic cells., J. Bone Miner. Metab., 28, 418-423 (2010).
  13. M. Yamaoka, M. Ishizuka, K. Ishihama, M. Takahashi, M. Takahashi, H. Yamada, Y. Teramoto, K. Yasuda, T. Shiba, T. Uematsu, K. Furusawa, Bone formation without lamina dura in the middle-aged and elderly: possible dependence on enamel., Clinical Interventions in Aging, 5, 37-43 (2010).
  14. K. Morita, K. Doi, T. Kubo, R. Takeshita, S. Kato, T. Shiba, Y. Akgagawa, Enhanced initial bone regeneration with inorganic polyphosphate-adsorbed hydroxyapatite., Acta Biomaterialia, 6, 2808-2815 (2010).
  15. Q. Yuan, T. Kubo, K. Doi, K. Morita, R. Takeshita, S. Katoh, T. Shiba, P. Gong, Y. Akagawa, Effect of combined application of bFGF and inorganic polyphosphate on bioactivities of osteoblasts and initial bone regeneration., Acta Biomaterialia, 5, 1716-1724 (2009).
  16. Y. Kawazoe, S. Katoh, Y. Onodera, T. Kohgo, M. Shindoh, T. Shiba, Activation of the FGF signaling pathway and subsequent induction of mesenchymal stem cell differentiation by inorganic polyphosphate., Int. J. Biol. Sci., 4, 37-47 (2008).
  17. M. Yamaoka, T. Uematsu, T. Shiba, T. Matsuura, Y. Ono, M. Ishizuka, H. Naramoto, M. Takahashi, M. Sugiura-Tomita, K. Iguchi, S. Yamashita, K. Furusawa, Effect of inorganic polyphosphate in periodontitis in the elderly., Gerodontology., 25, 10-17 (2008).
  18. Y. Hacchou, T. Uematsu, O. Ueda, Y. Usui, S. Uematsu, M. Takahashi, T. Uchihashi, Y. Kawazoe, T. Shiba, S. Kurihara, M. Yamaoka, K. Furusawa., Inorganic polyphosphate: a possible stimulant of bone formation., J. Dent. Res., 86, 893-897 (2007).
  19. P. McInerney, T. Mizutani, T. Shiba, Inorganic polyphosphate interacts with ribosomes and promotes translation fidelity in vitro and in vivo. Mol. Microbiol. 60, 438-447 (2006).
  20. H. Itoh, Y. Kawazoe, T. Shiba, Enhancement of protein synthesis by an inorganic polyphosphate in an E. coli cell-free system., J. Microbiol. Methods. 64, 241-249 (2006).
  21. H. Zhang, N. N. Rao., T. Shiba, A. Kornberg, Inorganic polyphosphate in the social life of Myxococcus xanthus: Motility, development, and predation., Proc. Natl. Acad. Sci. USA 102, 13416-13420 (2005).
  22. T. Shiba, H. Itoh, A. Kameda, Y. Kawazoe, K. Kobayashi, T. Noguchi, Polyphosphate:AMP phosphotransferase as a polyphosphate dependent nucleoside monophosphate kinase in Acinetobacter johnsonii 210A., J. Bacteriol., 187, 1859-1865 (2005).
  23. H. Itoh and T. Shiba, Polyphosphate synthetic activity of polyphosphate:AMP phosphotransferase in Acinetobacter johnsonii 210A., J. Bacteriol. 186, 5178-5181 (2004).
  24. Y. Kawazoe, T. Shiba, R. Nakamura, A. Mizuno, K. Tsutsumi, T. Uematsu, M. Yamaoka, M. Shindoh, T. Kohgo, Induction of calcification in MC3T3-E1 cells by inorganic polyphosphate., J. Dent. Res. 83, 613-618 (2004).
  25. T. Shiba, Y. Takahashi, T. Uematsu, Y. Kawazoe, K. Ooi, K. Nasu, H. Itoh, H. Tanaka, M. Yamaoka, M. Shindoh, T. Kohgo, Effect of Inorganic Polyphosphate on Periodontal Regeneration., Key Engineering Materials, 254-256, 1119-1122 (2004).
  26. T. Shiba, D. Nishimura, Y. Kawazoe, Y. Onodera, K. Tsutsumi, R. Nakamura, M. Ohshiro, Modulation of mitogenic activity of fibroblast growth factors by inorganic polyphosphate., J. Biol. Chem. 278, 26788-26792 (2003).
  27. K. Ishige, T. Hashimoto, T. Shiba, T. Noguchi, Novel method for enzymatic synthesis of CMP-NeuAc., Biosci. Biotechnol. Biochem., 65, 1736-1740 (2001).
  28. A. Kameda, T. Shiba, Y. Kawazoe, Y. Satoh, Y. Ihara, K. Ishige, T. Noguchi, A novel regeneration system using polyphsophate-AMP phosphotransferase and polyphosphate kinase., J. Biosci. Biotechnol. 9, 557-563 (2001).
  29. K, Tsutsumi, M. Munekata, T. Shiba, Involvement of inorganic polyphosphate in expression of SOS genes., Biochim. Biophys. Acta., 1493, 73-81 (2000).
  30. T. Miyake, T. Shiba, A. Kameda, Y. Ihara, M. Munekata, K. Ishige, T. Noguchi, The gene for exopolyphosphatase of Pseudomonas aeruginosa. DNA Research, 6, 103-108 (1999).
  31. T. Noguchi, T. Shiba, Use of Escherichia coli polyphosphate kinase for oligosaccharide synthesis., Biosci. Biotechnol. Biochem., 62, 1594-1596 (1998).
  32. K. Ishige, T. Shiba, T. Noguchi, The polyphosphate kinase gene of Pseudomonas aeruginosa. DNA research, 5, 157-162 (1998).
  33. T. Shiba, K. Tsutsumi, H. Yano, Y. Ihara, A. Kameda, T. Kan, H, Takahashi, M. Munekata, N. N. Rao, A. Kornberg, Inorganic polyphosphate and the induction of rpoS expression., Proc. Natl. Acad. Sci. USA 94, 15146-15152 (1997).
  34. H. Wurst, T. Shiba, A. Kornberg, The gene for a major exopolyphosphatase of Saccharomyces cerevisiae., J. Bacteriol. 177,898-906 (1995).