1. 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"]
Moscow, 2011/2012

2. 1. Summary of the innovative project (hereinafter – the Project)
General description of the Project Prospects of the elaboration
[Description of the problem to which the Project is addressed: Detection and differential  Synthesis of nanoscale UNP in the form of nanocrystals NaYF4 ranging in size from 10 to 100 nm, doped
diagnosis of abnormal tissue structures, such as tumor foci, as well as the exact location of with elements of the trivalent lanthanide Yb3+ and Tm3+ (or Yb3+ and Er3+) with the corresponding molar
their spatial boundaries in the operating field in real time during surgery, laparoscopic and 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.
endoscopic procedures, and a number of medical diagnostic procedures and research is one
of the most important and unresolved medical problems.] [Technical parameters of the final product of the project
[The essence of development (the description of the produced product (technology) and its  2. Achieving stability of UCNP-containing colloids in water and buffer systems (modification of the
characteristics): A new real-time intraoperative diagnostics technology that allows a clear surfaces of nanocrystals using amphiphilic polymers, with the replacement of surface residues oleic acids,
distinction between pathological lesions, bordering with healthy and inflamed tissues, as well as well as by encapsulation of UCNP in biocompatible polymers, and / or PEG-coated surfaces and
as hidden in the thickness of sediment layers and under inflammatory effusions, is proposed. proteins).
 3. Development of the targeting agent (antibody, mini-atibody or peptide) and universal platform for UCNP
Abnormal tissues are marked (on the basis of antigenic determinants) using targeted bioconjugation on the base of high affinity pair of proteins barnase:barstar as a molecular linker of strong
fluorescent molecular probes with unique photophysical properties - the ability to respond binding.
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  4. Engineering solutions: design and creation of laboratory model of the optical sensing, or optical
represent an immunospecific contrasting agent. Glow of BioNAF can be observed directly scanner, development and demonstration of the system complex for the excitation and detection of the
and registered in the thickness of tissues to a depth of 1 cm with a portable tomograph.] 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).
[The technological focus of the Project: biomedical technology, aimed at modernization of  5. Precise localization and differentiation of healthy and pathological tissues in animal models and,
surgical, including laparoscopic and endoscopic, practice.] 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.]
The stage of the project, on which this application is submitted
[Technical parameters of the product achieved at the time with indication on the presence of evidential material
 [The stage number in accordance with the Grant policy of the Foundation: this project
refers to the sowing stage (stage 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
The dynamics of the Project to date 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%
 [What has been done to address the issue (to create a product, technology): I. The against 0.1% in the case of UCNP with an average grain size of 30 nm. Optimization of conditions for the
technological basis: 1) the basic technology of synthesis, surface modification of the synthesis of nanocrystals will provide better quality and, consequently, an improved CC. The Act.
proposed luminescent materials is created, 2) the basic principles of bioconjugation of • Stability of UCPN-containing colloids in biocompatible buffers confirmed in long-term follow-up (one month)
upconverting nanophosphors (UCNP) to target biomolecules are developed, and 3) the for crystals with a diameter of 100 nm. The Act.
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. • Bioconjugates of UCNP modified by amphiphilic polymers, with a mini-antibodies to antigens of cancer
II. Project team: a workable interdisciplinary team, assembled for the Project, had cells carried HER2/neu, were synthesized; the specificity of binding fof BioNAF to this receptor was
demonstrated its effectiveness. III. The purpose and ways of solution: planned objective confirmed in the cell culture in vitro. The Act.
of the program and innovative ways to achieve this goal are choosen. IV. Support: the • The laboratory prototype of the optical scanner for deep tissue probing and detection of the luminescence
support of industrial partners interested in commercial output of this Project is UCNP/BioNAF was developed. Experiments on the numerical modeling of the optical properties of
approved.] biological tissues are performed. Layout.]
The current state [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.]
 [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 Plan to achieve
the stated objectives, so that the technical risks of the project can be considered to be  [The critical scientific and technological uncertainty, which will need to be addressed during the
minimized. We demonstrate the synthesis of UCNP doped with thulium and erbium research:achieving of stability of UCPN-containing colloids, suppressive effect of external biological
ions. 2. Versatile diagnostics of the basic physical and optical properties of the environment on the luminescence of UCNP/BioNAF, decrease of the intensity of the light source in the
synthesized UCNP has been carried out. The results were published, recent data is thickness of a turbid biological tissues.]
prepared for publication. 3. The stability of surface-modified UCNP in a biologically
compatible buffer solutions is confirmed. 4. Universal platform of bioconjugation  [Planned R & D stages and intermediate targets (functional and quantitative)
reaction on the base of high affinity molecular pair of barnase: barstar has been  I. (1). Synthesis of UCNP, based on Tm using new techniques. (2). Preparation of stable aqueous
demonstrated earlier in the inter-university research collaboration (Macquarie - IBCh). solutions based on the UCNP. (3). Numerical simulation and fabrication of a bio. phantoms for testing the
The results were published. 5. Activity of the targeting agent was demonstrated. The methodology of optical sensing with UCNP . (4). Designing of Lab. Layout of Opt. sensing. II. (1).
results were published. 6. Reliable results have been obtained on the estimation of the Diagnostics of Tm-based UCNP optical and physical properties. (2). Obtaining a stable buffer solutions
optical fluorescence signal from the inclusion of UCNP in a biological phantom, which based on the UCNP. (3). Creating a laboratory model of the optical sensing (optical tomography). III. (1).
makes the stated depth of the optical sensing (1 cm) achievable. 7. Basic laboratory Bioconjugation of biomolecules to the surface of UCNP (demonstration). IV. (1). Development of a
models of optical tomography were designed and tested. 8. Preliminary testing of the 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
basic ideas of immunochemical analysis using whole blood was performed.] targeted with antibodies towards aimed antigen.]
2

3. 2. Target market and competition
Needs of the consumers Development of similar products
 [The problem, the lack of existing solutions and overcoming. Inability to translation data from the preoperative stage to the  [1. Methods of synthesis and surface modification of
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 upconversion nanophosphors 2. Methods and solutions
autofluorescence, as well as by reducing the toxicity of contrast agents, providing the binding specificity of the probes with the for optical sensing and optical tomography, and 3.
targeted structures and simultaneous recording of signals from the surface of tissues and from their thickness.]
Solutions for specific imaging of biological tissue with the

[The market appeal of the target product. 1. Luminescent nanocrystals that have an unique ability to convert the exciting infrared
use of fluorescent probes.]
radiation into the luminescence in the visible and infrared ranges: solution of the problem of tissue autofluorescence, a clear  [This project proposes the union of these methodological
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 approaches into the complex technology. The exact
combination of UCNP an targeting biomolecules. 4. The unique engineering solution for the simultaneous excitation and definition of the target niche.]
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  [Of the product will be demanded by the market at the
about 0.1 mm, with a deep tomography - 1 mm. 2. The possibility of simultaneous information obtaining not only from the surface, time of commercialization, taking into account the forecast
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.] of development of technologies due to the precise
definition of its target niche - intraoperative diagnostics
Solutions existing on the market with a combination of direct imaging and optical
tomography of pathological foci.]
Name of the model/product Stage [Visualization of [Depth of [The [Visualization Price / cost of ownership,,
the pathological sensing of accuracy of + [USD]
focus] the biological localization tomography]
tissue] (estimation)]
[Several
[NightOWL II – LB 983 NC 100 (Berthold Tech.)] [On market] [No] [4 sm] [No] [$500 000]
mm]
[Fluorescent intraoperative [In
[Yes] [1÷4 sm] [0,1 mm] [No] [$200 000]
sensing based on the protoporphyrins] development]
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
The segment(s) of market for which the product is oriented
technology.]
Potential consumers of the product of the Project
[Participants of of the medical services (diagnostics) market; / Research Institutions (biomedical
 [Russia and the CIS countries, USA, Europe, Australia] [Products and technology of research); / The mass consumer (disposable biochips for rapid immunochemical tests).]
medical purpose]
Market appraisal [2012] [2017] [2020]
In qualitative terms, [consumers, a number of medical facilities] [1000] [2000] [5000]
World market
In monetary terms, [bn USD] [1] [5] [7]
Domestic In qualitative terms, [consumers, a number of medical facilities] [1000] [2000] [5000]
Примечание:
market а) если применимо terms, [bn USD] [Источник 1], [Источник 2], [Источник 3]
In monetary Источники: [200] [230] [300] 3

4. 3. The technology and intellectual property
Existing patents
Description of the innovation
 Russian patents
[The technology of direct visualization and optical imaging based on use of
nanoscale upconverting nanophosphors and their bioconjugates (BioNAF) will ‒ [Nechaev AV Shilov, IP, Panas AI, Rumyantseva VD, Shevelyova
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 EV, MironovAF, Markushev VM, Baryshnikov AYu, "Ytterbium
operated/examined organ) and optical tomography (at the depth of 1 cm).] complexes of tetrapirazolilporphins as fluorescent labels for
[The main advantages of the technology 1. Luminescent nanocrystals that have diagnostics of malignant neoplasms," Russian patent number
an unique ability to convert the exciting infrared radiation into the luminescence in 237,099. Publ. 10/11/2009. - Bull. Picture. Number 31. - C. 657.]
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  PCT
with the targeting molecules: immunomorphological (most specific) staining of
tissues and cells in situ. 3. Universal platform for bioconjugation: the ability to ‒ [V. V. Sokovikov, A. V. Zvyagin, B. Sandnes, “A method of the
create virtually any combination of UCNP an targeting biomolecules. 4. The unique
engineering solution for the simultaneous excitation and detection of luminescence, automatic, long-range identification of an object, and its
a combination of direct visualization and deep optical tomography.] realisation means of a retro-emission device and an optical
Description of the scientific and technological novelty reader”, filed 21 September 2010, Federal Office of Intellectual
[The novelty of the technology compared to existing developments is the unique Property, Patents and Trade Marks, Moscow, Russian
engineering solution of the complex for excitation and detection of luminescence, Federation, file No. 2010138851.
which allows the simultaneous visualization of the pathological focus and depth-
sensing of tissues, as well as the integration into the complex decision the ‒ A. V. Zvyagin, V. V. Sokovikov, S. A. Ermilov, I. I. Krokhin, “A
processes of synthesis and modification and production of UCNP and their method of location, tracking, and reading of a fluorescent matrix
bioconjugates with targeting molecules (BioNAF).]
code, and its embodiment”, No. 93168, priority date 21 July
[Thanks to the innovations it will be achieved significant changes in the 2009, registered 20 April 2010, Federal Office of Intellectual
technologies currently used (increase in accuracy and speed of of diagnostics,
reducing the trauma and the volume of surgical operations), as well as cost Property, Patents and Trade Marks, Moscow, Russian
savings of diagnosis and treatment of patients.] Federation.]
[Key competitive advantages of the projected product derived from the application
of scientific and technological innovation:1. The unprecedented accuracy of the ‒ Planned patents
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  Russian patents, then - the PCT
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 ‒ [1. The method of optical sensing based on use of delayed
a wide range of target antigens. 4. Non-toxic nature of BioNAF. 5. Economical and registration scheme 2012/2013 . 2. The synthetic procedure for
operational benefits. ]
registration of signals in the intraoperative diagnostics
[Global competitiveness and / or a positive impact on import substitution. The
proposed technology is competitive in the domestic and global markets and can 2012/2013 .]
have a positive impact on import substitution.]  PCT
[Impact on key product chain of the economical sector. Preparations of BioNAF ‒ [The method of optical reading (retroemission) for
can replace a significant segment of contrast agents used for medical purposes. immunochemical analysis, 2013/2014]
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.]
20.07.12

5. 4. Project Team and co-investor
A brief résumés of key team members of the Project
Plan 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.]
20.07.12 5

6. 5. Roadmap and financial plan of the Project
1. Aggregative plan of the Project’s development:
– Commercialization Plan (in the future of the next 5 years). There are three main directions:
– One-time sales of optical sensing system complexes or optical tomographs.
– Regular supply preparations on the basis of the physiological BioNAF;
– Production and sales of biochips for the express diagnostics on the basis of BioNAF.
- Plan to attract investments. Three main strategies are developed:
– Consolidation of joint scientific and technical activity with co-investor of this project, in particular the NTC "Amplitude.“
– Promotion of the investment program as part of the Russian Venture Company (RVC).
– 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:
- Studies of pharmacokinetics and pharmacodynamicsas well as toxicological assessment of BioNAF;
- 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.
- 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:
- 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.
- Optical sensing method based on use of delayed registration scheme.
- 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.
07/20/12 6

7. Current status (Completed The stage on which the grant
Key actions Subsequent stages
activities) is requested
ПРЕДВАРИТЕЛЬНЫЙ ПЛАН-ГРАФИК РЕАЛИЗАЦИИ ПРОЕКТА year
I,
Terms of implementation
year
II,
year
III,
year
IV,
year
I,
year
II,
year
III,
year
IV,
year
I,
year
II,
year
III, IV,
year
Research and development
1. [Synthesis of (1) and the optimization of the photophysical properties (2) ща upconverting
- - - 2 - - - - - - -
nanophosphors with the required characteristics (size 10 - 100 nm, NaYF4:Yb:Tm)]
1
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]
˅ + +
Конфиденциально