Moscow, 2011/2012

PRESENTATION, attached to the application on the grant from “Skolkovo” Foundation

Name of innovation project: [NOVEL LUMINESCENT NANOMATERIALS AND THEIR APPLICATIONS IN BIOMEDICAL TECHNOLOGY]

Name of the organization: [LLC "Mithrel-Lumitech"]

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1. Summary of the innovative project (hereinafter – the Project)

General description of the Project

[Description of the problem to which the Project is addressed: Detection and differential diagnosis of abnormal tissue structures, such as tumor foci, as well as the exact location of their spatial boundaries in the operating field in real time during surgery, laparoscopic and endoscopic procedures, and a number of medical diagnostic procedures and research is one of the most important and unresolved medical problems.]

[The essence of development (the description of the produced product (technology) and its characteristics): A new real-time intraoperative diagnostics technology that allows a clear distinction between pathological lesions, bordering with healthy and inflamed tissues, as well as hidden in the thickness of sediment layers and under inflammatory effusions, is proposed.

Abnormal tissues are marked (on the basis of antigenic determinants) using targeted fluorescent molecular probes with unique photophysical properties - the ability to respond prolonged luminescence in the visible and IR radiation on the excitation of the near-IR spectrum. These probes are bioconjugates of upconverting nanophosphors (BioNAF) and represent an immunospecific contrasting agent. Glow of BioNAF can be observed directly and registered in the thickness of tissues to a depth of 1 cm with a portable tomograph.]

[The technological focus of the Project: biomedical technology, aimed at modernization of surgical, including laparoscopic and endoscopic, practice.]

The stage of the project, on which this application is submitted

[The stage number in accordance with the Grant policy of the Foundation: this project refers to the sowing stage (stage 1).]

The dynamics of the Project to date

[What has been done to address the issue (to create a product, technology): I. The technological basis: 1) the basic technology of synthesis, surface modification of the proposed luminescent materials is created, 2) the basic principles of bioconjugation of upconverting nanophosphors (UCNP) to target biomolecules are developed, and 3) the basic laboratory optical tomography system is designed, and 4) the first steps in the numerical modeling and quantifying the sensitivity of and probing depth are performed. II. Project team: a workable interdisciplinary team, assembled for the Project, had demonstrated its effectiveness. III. The purpose and ways of solution: planned objective of the program and innovative ways to achieve this goal are choosen. IV. Support: the support of industrial partners interested in commercial output of this Project is approved.]

The current state

[Current state of research carried out by the team and the parameters of the existing results, samples: 1. To date, significant progress achieved in advancing the project to the stated objectives, so that the technical risks of the project can be considered to be minimized. We demonstrate the synthesis of UCNP doped with thulium and erbium ions. 2. Versatile diagnostics of the basic physical and optical properties of the synthesized UCNP has been carried out. The results were published, recent data is prepared for publication. 3. The stability of surface-modified UCNP in a biologically compatible buffer solutions is confirmed. 4. Universal platform of bioconjugation reaction on the base of high affinity molecular pair of barnase: barstar has been demonstrated earlier in the inter-university research collaboration (Macquarie - IBCh). The results were published. 5. Activity of the targeting agent was demonstrated. The results were published. 6. Reliable results have been obtained on the estimation of the optical fluorescence signal from the inclusion of UCNP in a biological phantom, which makes the stated depth of the optical sensing (1 cm) achievable. 7. Basic laboratory models of optical tomography were designed and tested. 8. Preliminary testing of the basic ideas of immunochemical analysis using whole blood was performed.]

Prospects of the elaboration

Synthesis of nanoscale UNP in the form of nanocrystals NaYF4 ranging in size from 10 to 100 nm, doped with elements of the trivalent lanthanide Yb3+ and Tm3+ (or Yb3+ and Er3+) with the corresponding molar ratio of 18% and 2% to the achievement of phase states of a crystal with a hexagonal crystal lattice (beta phase), which has an unprecedented high conversion coefficient.

[Technical parameters of the final product of the project

2. Achieving stability of UCNP-containing colloids in water and buffer systems (modification of the surfaces of nanocrystals using amphiphilic polymers, with the replacement of surface residues oleic acids, as well as by encapsulation of UCNP in biocompatible polymers, and / or PEG-coated surfaces and proteins).

3. Development of the targeting agent (antibody, mini-atibody or peptide) and universal platform for UCNP bioconjugation on the base of high affinity pair of proteins barnase:barstar as a molecular linker of strong binding.

4. Engineering solutions: design and creation of laboratory model of the optical sensing, or optical scanner, development and demonstration of the system complex for the excitation and detection of the luminescence named "IOVIZOR", which allows reveal the pathological foci with a diameter of 0.1 mm, labeled by probes of BioNAF on the surface of organs as well as in their thickness (to a depth of 1 cm).

5. Precise localization and differentiation of healthy and pathological tissues in animal models and, subsequently, in medical diagnostic research, especially in surgery, laparoscopic and endoscopic practice, as well as the detection of diseases of the skin and mucous membranes.]

[Technical parameters of the product achieved at the time with indication on the presence of evidential material

1. Mastered the synthesis of NaYF4: Yb3 +: Er3 + , the first experiments on the synthesis of NaYF4: Yb3 +: Tm3 + were carried out. The resulting samples vary in size, dimensional dispersion, as well in the nanocrystals quality. Depending on this, the optical properties of UCNP vary significantly . Conversion coefficient (CC) of the UCNP remains low, essentially yielding to micron-sized nanophosphors' CC, as 4% against 0.1% in the case of UCNP with an average grain size of 30 nm. Optimization of conditions for the synthesis of nanocrystals will provide better quality and, consequently, an improved CC. The Act.

2. Stability of UCPN-containing colloids in biocompatible buffers confirmed in long-term follow-up (one month) for crystals with a diameter of 100 nm. The Act.

3. Bioconjugates of UCNP modified by amphiphilic polymers, with a mini-antibodies to antigens of cancer cells carried HER2/neu, were synthesized; the specificity of binding fof BioNAF to this receptor was confirmed in the cell culture in vitro. The Act.

4. The laboratory prototype of the optical scanner for deep tissue probing and detection of the luminescence UCNP/BioNAF was developed. Experiments on the numerical modeling of the optical properties of biological tissues are performed. Layout.]

[Planned date of implementation of the Project and the beginning of commercialization: Start of the Project - October 2012, the beginning of the commercialization of of the Project - the third quarter of 2012.]

Plan to achieve

[The critical scientific and technological uncertainty, which will need to be addressed during the research:achieving of stability of UCPN-containing colloids, suppressive effect of external biological environment on the luminescence of UCNP/BioNAF, decrease of the intensity of the light source in the thickness of a turbid biological tissues.]

[Planned R & D stages and intermediate targets (functional and quantitative)

I. (1). Synthesis of UCNP, based on Tm using new techniques. (2). Preparation of stable aqueous solutions based on the UCNP. (3). Numerical simulation and fabrication of a bio. phantoms for testing the methodology of optical sensing with UCNP . (4). Designing of Lab. Layout of Opt. sensing. II. (1). Diagnostics of Tm-based UCNP optical and physical properties. (2). Obtaining a stable buffer solutions based on the UCNP. (3). Creating a laboratory model of the optical sensing (optical tomography). III. (1). Bioconjugation of biomolecules to the surface of UCNP (demonstration). IV. (1). Development of a promising targeted delivery agent. (2). Demonstration of Opt. sensing, biolog.phantoms and optical tomograph. (3) Testing of the model of immunochemical analysis with use of bioconjugated UCNP targeted with antibodies towards aimed antigen.]

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2. Target market and competition

Needs of the consumers [The problem, the lack of existing solutions and overcoming. Inability to translation data from the preoperative stage to the

intraoperative diagnostics: the need to create system for navigation in the operating field on the basis of differential contrast study of tissues. Solutions based on the use of fluorescent probes can be significantly improved by supression of tissue autofluorescence, as well as by reducing the toxicity of contrast agents, providing the binding specificity of the probes with the targeted structures and simultaneous recording of signals from the surface of tissues and from their thickness.]

[The market appeal of the target product. 1. Luminescent nanocrystals that have an unique ability to convert the exciting infrared

radiation into the luminescence in the visible and infrared ranges: solution of the problem of tissue autofluorescence, a clear visualization of the boundaries of labeled foci. 2. Bioconjugation of UCNP with the targeting molecules: immunomorphological (most specific) staining of tissues and cells in situ. 3. Universal platform for bioconjugation: the ability to create virtually any combination of UCNP an targeting biomolecules. 4. The unique engineering solution for the simultaneous excitation and detection of luminescence, a combination of direct visualization and deep optical tomography.

[Demand motivation. 1. The highest accuracy of the localization of pathological foci (spatial resolution for direct visualization is about 0.1 mm, with a deep tomography - 1 mm. 2. The possibility of simultaneous information obtaining not only from the surface, but from the depths of the body, decrease of the number of cuts and reduction of volumes of operations. 3. Adaptiveness of the solution to detect a wide range of target antigens. 4. Non-toxic nature of BioNAF. 5. Economical and operational benefits.]

Development of similar products [1. Methods of synthesis and surface modification of

upconversion nanophosphors 2. Methods and solutions for optical sensing and optical tomography, and 3. Solutions for specific imaging of biological tissue with the use of fluorescent probes.]

[This project proposes the union of these methodological approaches into the complex technology. The exact definition of the target niche.]

[Of the product will be demanded by the market at the time of commercialization, taking into account the forecast of development of technologies due to the precise definition of its target niche - intraoperative diagnostics with a combination of direct imaging and optical tomography of pathological foci.]

Solutions existing on the market

Name of the model/product Stage [Visualization of the pathological

focus]

[Depth of sensing of the biological

tissue]

[The accuracy of localization(estimation)]

[Visualization + tomography]

Price / cost of ownership,, [USD]

[NightOWL II – LB 983 NC 100 (Berthold Tech.)] [On market] [No] [4 sm][Several mm]

[No] [$500 000]

[Fluorescent intraoperativesensing based on the protoporphyrins]

[In development]

[Yes] [1÷4 sm] [0,1 mm] [No] [$200 000]

Примечание: а) если применимо Источники: [Источник 1], [Источник 2], [Источник 3]

Market appraisal [2012] [2017] [2020]

World marketIn qualitative terms, [consumers, a number of medical facilities] [1000] [2000] [5000]

In monetary terms, [bn USD] [1] [5] [7]

Domestic market

In qualitative terms, [consumers, a number of medical facilities] [1000] [2000] [5000]

In monetary terms, [bn USD] [200] [230] [300]

The scheme of commercialization[One-time sales of system complexes of optical sensing or optical tomographs. Sales of drugs BioNAF and biochips for rapid diagnostics based on the BioNAF. Sales of licenses for the use of technology.]

The segment(s) of market for which the product is oriented

[Russia and the CIS countries, USA, Europe, Australia] [Products and technology of medical purpose]

Potential consumers of the product of the Project [Participants of of the medical services (diagnostics) market; / Research Institutions (biomedical

research); / The mass consumer (disposable biochips for rapid immunochemical tests).]

Existing patents Russian patents

‒ [Nechaev AV Shilov, IP, Panas AI, Rumyantseva VD, Shevelyova EV, MironovAF, Markushev VM, Baryshnikov AYu, "Ytterbium complexes of tetrapirazolilporphins as fluorescent labels for diagnostics of malignant neoplasms," Russian patent number 237,099. Publ. 10/11/2009. - Bull. Picture. Number 31. - C. 657.]

PCT

‒ [V. V. Sokovikov, A. V. Zvyagin, B. Sandnes, “A method of the automatic, long-range identification of an object, and its realisation means of a retro-emission device and an optical reader”, filed 21 September 2010, Federal Office of Intellectual Property, Patents and Trade Marks, Moscow, Russian Federation, file No. 2010138851.

‒ A. V. Zvyagin, V. V. Sokovikov, S. A. Ermilov, I. I. Krokhin, “A method of location, tracking, and reading of a fluorescent matrix code, and its embodiment”, No. 93168, priority date 21 July 2009, registered 20 April 2010, Federal Office of Intellectual Property, Patents and Trade Marks, Moscow, Russian Federation.]

‒ Planned patents Russian patents, then - the PCT

‒ [1. The method of optical sensing based on use of delayed registration scheme 2012/2013 . 2. The synthetic procedure for registration of signals in the intraoperative diagnostics 2012/2013 .]

PCT‒ [The method of optical reading (retroemission) for

immunochemical analysis, 2013/2014]

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3. The technology and intellectual property

Description of the innovation[The technology of direct visualization and optical imaging based on use of nanoscale upconverting nanophosphors and their bioconjugates (BioNAF) will allow revealing and identifying abnormal structures immediately in the course of surgery or diagnostic test, in real time by direct visualization (on the surface of the operated/examined organ) and optical tomography (at the depth of 1 cm).]

[The main advantages of the technology 1. Luminescent nanocrystals that have an unique ability to convert the exciting infrared radiation into the luminescence in the visible and infrared ranges: solution of the problem of tissue autofluorescence, a clear visualization of the boundaries of labeled foci. 2. Bioconjugation of UCNP with the targeting molecules: immunomorphological (most specific) staining of tissues and cells in situ. 3. Universal platform for bioconjugation: the ability to create virtually any combination of UCNP an targeting biomolecules. 4. The unique engineering solution for the simultaneous excitation and detection of luminescence, a combination of direct visualization and deep optical tomography.]

Description of the scientific and technological novelty[The novelty of the technology compared to existing developments is the unique engineering solution of the complex for excitation and detection of luminescence, which allows the simultaneous visualization of the pathological focus and depth-sensing of tissues, as well as the integration into the complex decision the processes of synthesis and modification and production of UCNP and their bioconjugates with targeting molecules (BioNAF).]

[Thanks to the innovations it will be achieved significant changes in the technologies currently used (increase in accuracy and speed of of diagnostics, reducing the trauma and the volume of surgical operations), as well as cost savings of diagnosis and treatment of patients.]

[Key competitive advantages of the projected product derived from the application of scientific and technological innovation:1. The unprecedented accuracy of the localization of pathological foci (spatial resolution for the direct visualization is 0.1 mm, and for a deep tomography - 1 mm. 2. Ability to simultaneously obtain information not only from the surface, but from the depths of the body, reducing the number of cuts and volumes of operations. 3. Adaptability of the solutions to detect a wide range of target antigens. 4. Non-toxic nature of BioNAF. 5. Economical and operational benefits. ]

[Global competitiveness and / or a positive impact on import substitution. The proposed technology is competitive in the domestic and global markets and can have a positive impact on import substitution.]

[Impact on key product chain of the economical sector. Preparations of BioNAF can replace a significant segment of contrast agents used for medical purposes. Complexes "IOVIZOR" have significant advantages before other imaging methods (PET, CT, ultrasound). This may contribute to the stabilization / stagnation of market of drugs and devices for diagnostic contrast study of tissues.]

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4. Project Team and co-investor

A brief résumés of key team members of the ProjectPlan to attract investment

Attraction of financing in the current stage

LLC STC «Amplitude» (Moscow, Zelenograd)]

[Affiliation with a co-investor: The lack of affiliation or potential conflict of interests of the applicant with the staff of the Foundation, members of the Investment Committee, the persons involved in the decision on the proposal is declared.]

[Mechanism for making investments into the participant company: Direct investments - 2 million rubles, + supply of equipment, infrastructure, as well as work time of paid staff for a total of 3 million rubles.]

[Profile of a co-investor: the experience in implementation of similar projects, financial state of the co-investor. Diversified company engaged in the production, delivery and maintenance of measuring instruments, special laboratory equipment, methodology, metrology and software in the field of radiation control, nuclear medicine and petrophysics, as well as development and implementation of technology of automatic bar code identification based on the technology of retro-emission and UCPN use in biomedicine.]

[How and in what period the co-investor plans to return the investment: Return of investments is possible after the start of sales of complex "IOVIZOR" (presumably, in 1-2 years after the start of the Project.

[Consortium of private co-investors]: the total amount of investments is 7.5 millions rubles.

Attraction of financing during the subsequent stages of the Project

[Requested funding – 30 mln. rub. for one year.

[After completion of the current stage of the project it is assumed yield to self-sufficiency.]

1. Aggregative plan of the Project’s development:

– Commercialization Plan (in the future of the next 5 years). There are three main directions: 1) One-time sales of optical sensing system complexes or optical tomographs.

2) Regular supply preparations on the basis of the physiological BioNAF;

3) Production and sales of biochips for the express diagnostics on the basis of BioNAF. - Plan to attract investments. Three main strategies are developed:

1) Consolidation of joint scientific and technical activity with co-investor of this project, in particular the NTC "Amplitude.“

2) Promotion of the investment program as part of the Russian Venture Company (RVC).

3) We also consider the possibility of funding the next (2nd) stage of the project by Skolkovo Foundation. - R&D Plan. Upon completion of the Stage 1 the new technology platform that can persuade medical specialists to its high practical potential will be developed.

However, in order to bring the technology to successful commercialization, ie before introduction into clinical practice, the following major steps of further development of the program to be required:

1) Studies of pharmacokinetics and pharmacodynamicsas well as toxicological assessment of BioNAF;

2) Design and manufacturing of an ergonomic model of the transportable model of the optical tomograph;Testing of the main provisions of intraoperative imaging and optical sensing in animals.

3) Clinical trials of the methodology of "Iovizor." - The plan to protect the intellectual property. On the first stages of the the project it is planned immediately to patent the basic concepts of technology

immediately. In particular, we highlight the following key ideas that require patent protection as soon as possible:

1) A new synthetic approach to the intraoperative procedure that can provide a visualization of superficial pathological foci, marked with specially designed molecular probes BioNAF and optical sensing to a depth of 1 cm.

2) Optical sensing method based on use of delayed registration scheme.

3) Promotion of previously patented method of effective optical reading, called retroemission. This method patented in the Russian Federation, planned to be

used in the technology of immunochemical analysis with BioNAF. 2. Financial Plan of the Project:

- The requested amount of funds (with the proportion of by Skolkovo / co-investor). The total amount and proportion of deposits: Skolkovo - 22.5 million rubles. / Co-investors - 7.5 mln. rubles/ Applicant - no. Grant support for research projects under the program.

- The costs of the Project: 30 million rubles (Equipment-20% rent of premises - 4.8%, outsourcing - 9%, transport - 6.6% wage fund - 48%, materials and

components - 3%, the protection of intellectual property - 1.88%).

- Expected key financial results and the estimated term of their achievements. It is assumed sale of an experimental batch of UCNP in the 4th quarter of 2013

amounting to 200 thousand rubles. 04/13/23 6

5. Roadmap and financial plan of the Project

ПРЕДВАРИТЕЛЬНЫЙ ПЛАН-ГРАФИК РЕАЛИЗАЦИИ ПРОЕКТА

Конфиденциально

Key actionsCurrent status (Completed

activities)The stage on which the grant

is requestedSubsequent stages

Terms of implementation I,

yearII,

yearIII,

yearIV,

yearI,

yearII,

yearIII,

yearIV,

yearI,

yearII,

yearIII,

yearIV,

year

Research and development 1. [Synthesis of (1) and the optimization of the photophysical properties (2) ща upconverting

nanophosphors with the required characteristics (size 10 - 100 nm, NaYF4:Yb:Tm)] 1- - - 2 - - - - - - -

2. [Preparation colloids of UCNP stable in buffer solutions (experimental (1) and production (2) steps)] 1 - - - 2 - - - - - - -

3. [Preparation of bioconjugates of UCNP (BioNAF)] + + - - -

4. [Demonstration of the functionality of BioNAF with targeting molecules (antibodies)] ˅ +

5. [Creation of a model of the optical signal of UCNP at a depth of 1 cm in a turbid medium] ˅ +

6. [Creation of the demonstration layout of the optical tomograph] ˅ +

7. [Demonstration of immunochemical analysis with BioNAF] ˅ +

Marketing and adoption 1. [Demonstration of methods and technologies of "Iovizor" to health professionals, the

dissemination of information] ˅ +

2. [The consolidation of scientific and technical activity with co-investor of the Project, in particular, with NTC "Amplitude"] ˅ +

3. [Promotion of the investment program as part of the Russian Venture Company (RVC)] ˅ +

4. [Promotion of the investment program as part of the Russian Venture Company (RVC)] ˅ + +

Intellectual property

1. [Patenting of two key ideas of the Project in the Russian Federation, the promotion of patented technologies] + + +

2. [Patenting under the PCT in the U.S. and major industrialized countries]. - + +

Funding

1. [The initial development and pre-financing of the Project]+ +

2. [Funding for the project in stages I and II] ˅ + +