Technology offers portfolio

CNRS ONCOLOGY

16/02/2017 Emmanuelle Le Coz V2

83 Bd Exelmans, 75016 PARIS Tel + 33 (0) 1 40 51 00 90 Fax + 33 (0) 1 40 51 78 58 Email : frinnov@fist.fr – web : www.fist.fr S.A. au capital de 1 128 122€ R.C.S B 388 461 154 SIREN 388 461 154 00030 Code APE 6630Z TVA CEE FR 60 388 461 154

2 Table of content

Table of contents Part 1: Therapy ....................................................................................................................................................... 3

Part 1.1: Small molecules ........................................................................................................................................ 4

A potential first-in-class drug for the treatment of leukemia ................................................................................. 5

Targeted drugs for brain cancer .............................................................................................................................. 7

Platinum-based n-heterocylic carbenic complexes showing very efficient anti-cancer activity ........................... 10

Part 1.2: Antibodies .............................................................................................................................................. 12

LLT1 Antibodies with new functional properties .................................................................................................. 13

Part 1.3: Peptides .................................................................................................................................................. 14

Chimeric molecule involving oligomerized FasL extracellular domain ................................................................. 15

Tumor vaccine for immunotherapy of tumoral pathologies related to EBV latency infections ........................... 16

Novel recombinant lectins and applications for the detection of pathological state markers ............................. 18

Part 1.4: Biomarkers (targets) ............................................................................................................................... 20

New target involved in DNA replication for diagnosing and treating cancers ...................................................... 21

Part 2: Pronostic .................................................................................................................................................... 22

Development of Anti Trio antibody to detect Trio as a potential pronostic marker for cancer ........................... 23

Part 3: Theranostic ................................................................................................................................................ 24

A novel methodology to predict the response to HER2 bloking agent treatment in cancer ................................ 25

Part 4: Diagnostic .................................................................................................................................................. 27

Use of an inhibitory protein of cell migration as a diagnostic marker for metastastic progression ..................... 28

Mannose 6-phosphate receptor, new target for prostate cancer diagnosis ........................................................ 29

Novel recombinant lectins and applications for the detection of pathological state markers ............................. 30

CNRS Key figures ................................................................................................................................................... 32

Budget forecast for 2014 ................................................................................................................... 32

Personnel ........................................................................................................................................... 32

Organization ...................................................................................................................................... 32

International relations ....................................................................................................................... 32

Industrial relations ............................................................................................................................ 32

Awards ............................................................................................................................................... 32

3 Table of content

Part 1: Therapy

4 Table of content

Part 1.1: Small molecules

5 Table of content

A potential first-in-class drug for the

treatment of leukemia

CONTEXT The expression level of the cytosolic 5’-nucleotidase II (cN-II) is of crucial interest for patients treated with nucleoside analogue-based chemotherapy. Indeed, a high level of expression of cN-II mRNA in blasts is predictive of worse outcome in patients receiving cytarabine-based regimens (a well-known nucleoside analogue used to treat acute myeloid leukemia, AML). In addition, the inhibition of cN-II expression by short hairpin RNA was associated with the induction of apoptosis in human astrocytoma cells, suggesting that cN-II could be a therapeutic target in brain tumors. Finally, recent reports showed that hyperactive mutated cN-II in relapsed children with acute lymphoblastic leukemia, and treated with the anti-metabolite 6-mercaptopurine, was associated with a worse survival. In this context, cN-II has been shown to be an attractive molecular target for the development of novel drugs circumventing resistance to cytotoxic nucleoside analogues currently used for treating leukemia and other hematological malignancies.

TECHNICAL DESCRIPTION In light of the various structural, functional and regulatory properties of cN-II, the inventors considered cN-II as an attractive therapeutic target for developing different types of inhibitors that could interfere with protein function or regulation. Altering the enzymatic function of cN-II has an indirect effect on nucleotide pools and is supposed to enhance the therapeutic efficiency of cytotoxic nucleosides (by decreasing the competition with endogenous nucleotides). Indeed, numerous experimental evidences indicated that cN-II is interfering with anticancer treatment based on this class of therapeutics (nucleoside analogs) and therefore also governing the final outcome of patients. The main evidence is the relationship between the expression level of cN-II and the patient outcome. This feature is reinforced by the recent discovery of hyperactive mutants of cN-II that induce a similar response. The development of potent inhibitors against cN-II has been carried out and a family of small size compounds (designed for an oral delivery) has been characterized for their capability in blocking the cN-II enzymatic function and also in sensitizing the anti-proliferative effects of some cancer drugs using preclinical models.

DEVELOPMENT STAGE Multi-step chemical synthesis was been optimized, all intermediates and

final compounds were fully characterized. Synthesis has been carried out on mg to gram scale.

Reference 06043-01

Keywords [cN-II]; [5’-nucleotidases]; [nucleotides]; [Enzyme inhibitors]; [small molecule inhibitors]; [AML]; [Leukemia]; [cancer]. Status of Patent French priority patent application filed on October 1rst, 2013 and entitled “Inhibiteurs de 5’-nucléotidases et leurs utilisations thérapeutiques”

Inventors Suzanne PEYROTTES, Laurent CHALOIN & al. Commercial Status Exclusive or non-exclusive license Laboratory Institut des Biomolécules Max Mousseron (IBMM), UMR5247

6 Table of content

In vitro evaluation towards the recombinant purified cN-II (Nucleotidase activity inhibition assays). Two different in vitro assays were used for evaluating the inhibition of compounds. A colorimetric assays based on the dosage of inorganic phosphate produced by the reaction and a kinetic assay for the characterization of the inhibition mode and constant (Ki). Several compounds described in the invention have shown a potent inhibition of the cN-II activity (up to 90% of inhibition at 200 µM)

Cytotoxic activity validated on several tumor cell lines. Direct cytotoxicity was determined using cell survival assays on a large panel of cancer cell lines, showing IC50 in low micromolar range for the most potent inhibitors. Synergy with nucleoside analogues was shown for several associations.

Ex vivo PoC. The most potent cN-II inhibitors are also cytotoxic on human cancer cells obtained from patients with AML or CLL as shown by ex vivo incubation and cell survival analysis.

In vivo efficacy PoC. Initial experiments in a mouse model showed an antitumoral activity of cN-II inhibitors and a slight increase in the activity of associated nucleoside analogues.

BENEFITS The originality of the invention lies in the nature of the drug target (cytosolic and not membrane). No inhibitor of intracellular 5'-nucleotidases (cytosolic) is described to date for the treatment of human blood diseases (Acute lymphoblastic or myeloblastic leukemia and chronic lymphoid leukemia) for children and adult patients. The unique combination of the compounds of the invention, 5'-nucleotidases inhibitors, with cytotoxic nucleoside analogs known to date, increases the effectiveness of this drug class by several mechanisms: (1) by the intrinsic inhibition cN-II inducing a shorter cell survival ; (2) by increasing the intracellular concentration of phosphorylated forms (nucleotide) of associated nucleoside analogues, these entities being responsible for its antiproliferative activity; (3) by modulating some mechanisms of cellular resistance associated with the over expression or mutation of cN-II.

INDUSTRIAL APPLICATIONS These chemical compounds designed as inhibitors of 5'-nucleotidases can be used in the treatment of cancers, especially leukemia, alone or in combination with cytotoxic nucleoside analogs and / or nucleobases widely used in cancer chemotherapy, to potentiate their therapeutic effect. Eventually, it may be extended to other pathologies using nucleoside analogs

Et Centre d’étude d’agents Pathogènes et Biotechnologies pour la Santé (CPBS), UMR5236, Montpellier, France

7 Table of content

Targeted drugs for brain cancer

CONTEXT Medulloblastoma is a tumor of neuroectodermal origin occurring mainly in childhood. A subtype of pediatric medulloblastoma tumors is characterized by activated hedgehog (Hh) pathway, a pathway involved in the embryonic growth and development of organs. Ongoing clinical studies of inhibitors of Smoothened (Smo), a key receptor mediating the Hh pathway, have shown promising activity in Hh activated basal cell carcinoma and medulloblastoma. Nevertheless, as for most anticancer agents, resistance mechanisms have been reported in animal models of Hh-driven tumors and in a patient with medulloblastoma metastasis treated with a single agent (GDC-0449, Genentech) due to an acquired Smo mutation disrupting the ability of GDC-0449 to bind Smo.

TECHNICAL DESCRIPTION To allow the development of new Smo inhibitors, we have delineated a pharmacophore based on the structure of reference Smo antagonists. Then, we have selected a collection of compounds obtained from in silico screening of a virtual library (500,000 commercially available "drug-like" molecules) based on the design of this pharmacophore and identified 20 chemically unrelated ligands that we have ordered and tested in several Hh assays. Among them, we have identified a new lead called MRT-10 [1]. By structure-activity relationship, we have synthesized and characterized MRT-83 and MRT-72 that belong to novel families of molecules with original chemical structures that have been patented. [1] Manetti, F., Faure, H., Roudaut, H., Gorojankina, T., Traiffort, E., Schoenfelder, A., Mann, A., Solinas, A., Taddei, M., and Ruat, M., (2010) Virtual screening-based discovery and mechanistic characterization of the acylthiourea MRT-10 family as smoothened antagonists. Mol Pharmacol. 78(4): p. 658-65.

[2] Solinas, A., Faure, H., Roudaut, H., Traiffort, E., Schoenfelder, A., Mann, A., Manetti, F., Taddei, M., and Ruat, M., (2012) Acylthiourea, Acylurea and Acylguanidine Derivatives with potent Hedgehog inhibiting Activity. J Med Chem. 55(4): p. 1559-1571.

BENEFITS - Well identified target ; - Already identified and characterized proprietary leads acting on

Smoothened and its GDC-0449 resistant mutant ; - MRT-83 AND MRT-72 are among the more potent Smoothened

antagonists reported so far and are easy to synthesize ; - Developing second generation of Smo inhibitors for pediatric brain

tumors ; - MRT compounds may also be of interest for treating other

Hedgehog linked cancers.

Reference 01922-01 ; 02959-01 ;

04611-02 Keywords Hedgehog driven tumors ; Smoothened modulators ; Antagonist ; Drug resistance ; Pediatric oncology

Status of Patent 3 distinct patents families under priority of: 1) Priority patent application n° FR 08/02302 filed in April 24, 2008 and entitled “N-Acylthiourea and N-acylurea inhibitors of the Hedgehog protein signaling pathway” - International patent application n°PCT/FR2009/000442 filed in

April 16, 2009 : WO2009130422

- Validation of national phases in US, EP, CA, JP 2) Priority patent application n° FR 09/03654 filed in July 24, 2009 and entitled “Acylguanidine derivatives modulating the Hedgehog protein signaling pathway” - International patent application n°PCT/FR2010/000516 filed in July 19, 2010 - Validation of national phases in US, EP, CA, JP 3) Priority patent application n° FR 11/58519 filed in September 23, 2011 and entitled “Nouveaux composés modulateurs de la voie de signalisation des proétines Hedgehog, leurs formes marquées et applications” Inventors Martial RUAT et al. Commercial Status Exclusive license Laboratory Neurobiology and Development Laboratory (UPR3294), CNRS, in Gif-sur-Yvette (Paris area), France

8 Table of content

INDUSTRIAL APPLICATIONS The compounds could be mainly used as a drug for the treatment of tumors associated with a hyperactivation of the Hh protein signaling pathway (medulloblastoma, glioblastoma, basal cell carcinoma, rhabdomyosarcoma, pancreatic cancer etc.). Furthermore, in addition to cancer, these compounds can find applications for the treatment of a large number of diseases, notably diseases in which a blockage of the Hedgehog signaling pathway could be beneficial, such as diabetes.

DEVELOPMENT STAGE

Conceptual/ Pre-clinical Clinical Laboratory samples

Specificity and potency:

MRT-83 does not display antagonist activity towards the Wnt signalling pathway [1] and towards a panel of kinases, GPCR and transporters (unpublished data, primary screen done at CEREP).

MRT-83 and MRT-72 block the binding of Bodipy-Cyclopamine to rodent and human Smo (Table 1). MRT-83 displays a similar potency compared to GDC-0449 and LDE225 whereas MRT-72 is the most potent Smo antagonist described so far (Table 1). We have developed a tritiated form ([3H]MRT-72) which can be used as a selective and potent Smo radioligand.

Table 1. Comparison of Smo antagonists’ potency in different pharmacological assays. 1) Inhibition of the activity induced by 5 nM SHH (Biogen Idec) in SHH-light2 cells expressing the Gli1-dependent firefly luciferase. 2) Inhibition of the differentiation of C3H10T1/2 cells induced by 0.1μM SAG monitored by alkaline phosphatase (AP) activity. 3) Inhibition of SAG (0.01μM) induced rat granular cell precursor proliferation. 4) Inhibition of Bodipy-Cyclopamine binding (5 nM) to human Smo transfected in HEK293 cells. ND: Not Determined. IC50 are from 3 to 5 independent experiments.

Back up MRT molecules:

Interestingly, we have shown that the guanidine MRT-83 can be synthesized by the transformation of a thiourea or a urea function [2]. We have shown that the thiourea and urea analogs of MRT-83 are also very potent antagonists of Smo [2]. They represent back up analogs in our studies.

X

9 Table of content

Thus, we have synthesized and characterized MRT Smo antagonists that are among the most potent Smo antagonists known so far. At our knowledge, we are the first academic groups in Europe and at an international level to have proprietary Smo antagonists displaying subnanomolar potency. The emergence of resistance to Smo inhibitors has been only documented in one patient with medulloblastoma, the potential emergence of resistance has to be monitored more carefully in the ongoing clinical trials of Smo inhibitors in medulloblastoma, BCC and in other cancer. Therefore, evaluating the effects of MRT-83, MRT-72 and their backup analogs in mouse model of Hh driven medulloblastoma is of major interest.

10 Table of content

Platinum-based n-heterocylic carbenic

complexes showing very efficient anti-

cancer activity

CONTEXT Some platinum-based compounds are known to be efficient anti-cancer drugs. Among these compounds, the cisplatin is one of the more frequently used in the world. Nevertheless, this compound do have limitations: its scope of activity is restricted, some cancer cells have or can develop a resistance towards it, its side effects are numerous and its only method of delivery is through intravenous as it doesn’t show good solubility in water. Recently a new class of platinum-based compounds has been monopolizing scientists’ attention for its efficient anti-cancer activity: platinum-based N-heterocyclic carbenic complexes.

TECHNICAL DESCRIPTION In the present work, the inventors have discovered new platinum-based N-heterocyclic carbenic complexes that shows good cytosolubility and have developped a process to obtain said compounds. The process is a two-steps process through 1) the reaction of an imidazolium salt with a base, a platinum precursor and an amine forming a platinum-based N-heterocyclic carbenic complexes and 2) a second step which aims at modifying the first-step compound’s properties and especially its cytosolubility through a change of its ligand L.

DEVELOPMENT STAGE In vitro efficiency tests on breast cancer cells and brain cancer cells; Suitable process for multi-gram scale synthesis; Tumor-homing device under development.

BENEFITS The discovered compounds and its related process have numerous benefits. As for their anti-cancer activity, some show : Stronger activity than cisplatin for some compounds. As for the process, it offers: Limited number of sub-products; High yields; Simplicity and reconductibility; High modularity of ligands.

Reference 03978-01 -03

Keywords Platinum, cisplatin,

cancer, tumor, chemotherapy. Status of Patent French priority patent application FR1061227 filed on December the 23rd, 2010 and entitled « Nouveaux complexes carbéniques de platine et leur utilisation comme médicaments » French priority patent application FR1061228 filed on December the

23rd, 2010 and entitled « Procédé de préparation de complexes carbéniques » Inventors Gilles GUICHARD Edith CHARDON Stéphane BELLEMIN - LAPONNAZ Commercial Status Exclusive or non-exclusive licenses Laboratory Institut de Physique et Chimie des Matériaux de Strasbourg (IPCMS, UMR7504), Strasbourg, France Laboratoire de Chimie et Biologie des Membranes et Nanoobjets (CBMN, UMR5248), Pessac, France

11 Table of content

INDUSTRIAL APPLICATIONS As some of the protected compounds show better properties (high activity, cytosoluble) than cisplatin or other platinum-based anti-cancer compounds, the present invention aims at offering new candidates for cancer therapy. Some tumor-homing device aiming at targetting cancer cells are also under development.

12 Table of content

Part 1.2: Antibodies

13 Table of content

LLT1 Antibodies with new functional

properties

CONTEXT The discovery that Lectin-Like Transcript 1 (LLT1), also known as C-type lectin domain family 2 member D (CLEC2D), is a ligand for the CD161 receptor, also known as NKRP-1A, has led to explore several approaches for modulating the activities of cells of the immune system in the purpose of disease treatment. The inventors propose a new approach based on the use of monoclonal antibodies (Mab). Biological therapeutics are now available for the treatment of certain autoimmune and chronic inflammatory diseases and/or cancer. But there is still a need for alternative biological treatments which specifically target pathological tissues without affecting healthy tissue, thus resulting in less severe side effects, the said treatment may be used long-term. The current invention relates to these unmet needs amongst patients with cancer, and in those with autoimmune and chronic inflammatory diseases.

TECHNICAL DESCRIPTION The present invention relates to the characterization of monoclonal antibodies that are capable of specifically binding to LLT1, that block the interaction between LLT1 and its receptor CD161 and that have depleting properties in vitro and in vivo. The present invention also relates to polynucleotides encoding such antibodies and cells expressing such antibodies. These Mab have utility in the diagnosis and treatment of cancer and autoimmune/chronic inflammatory diseases, in which LLT1- and CD161- expressing cells play a role in disease pathogenesis.

DEVELOPMENT STAGE The inventors have developed murine, chimeric and humanized antibody

molecules that specifically bind to LLT1. They currently have 9 humanized antibodies with similar affinity with the murine anti-LLT1.

They have demonstrated using in vitro assays that the antibodies block the interaction between LLT1 and CD161, thereby stimulating the cytotoxicity and cytokine production of NK cells and inhibiting the proliferation and cytokine production of T cells.

They have demonstrated using in vitro assays that the antibodies facilitate the removal of LLT1-expressing cells by antibody-dependent cell-mediated cytotoxicity (ADCC). Preliminary pre-clinical tests in mice xenografted with human B cell lymphomas show in vivo efficacy of the anti-LLT1 antibody alone and in combination with Rituximab (anti-CD20, Mabthéra®).

Reference 03864-01

Keywords LLT1 Modulator; Monoclonal antibodies (Mab); CD161 Modulator, Cancer, autoimmune diseases

Status of Patent Priority patent application n° EP10167668, filed in June 29, 2010 - Priority patent application n° EP10168095, filed in July 01, 2010 - Priority patent application n° EP10169409 filed in July 13, 2010

WO201200994

EP2588498

US2013164299 Inventors Véronique BRAUD Commercial Status Research agreement, exclusive or non exclusive licenses

14 Table of content

Part 1.3: Peptides

15 Table of content

Chimeric molecule involving

oligomerized FasL extracellular domain

CONTEXT FasL (or CD95L, CD178) is a homotrimeric membrane-bound apoptosis inducer through the Fas receptor-triggered caspase-dependent apoptotic pathway. FasL also exists as a soluble form (sFasL) that is almost devoid of any apoptotic activity. We designed a polymeric form of

sFasL which is highly apoptotic, and can be used to eliminate Fas-expressing cells.

TECHNICAL DESCRIPTION This invention describes the obtention of the polymeric sFasL molecule (called pFasL), and of chimeric form of it consisting in a fusion to a protein able to interact with a component expressed on the surface of the Fas-expressing cells to be targeted. The pFasL consists in a chimera between a self-interacting domain of the human Leukemia Inhibitory Factor receptor gp190 and sFasL, separated by a peptidic linker. As a prototype, a CD80-pFasL chimera was engineered and analysed. CD80 is a ligand of CD28. CD28 is known as a membrane costimulating receptor for T-lymphocyte antigen-dependent activation, and is also expressed on the surface of tumoral B-lymphocytes such as myeloma and plasmocytoma cells. The CD80-pFasL chimera is polymeric and is a more potent apoptosis inducer than pFasL, due to the capacity of CD80 to interact with CD28 and thereby to enhance the apoptotic activity of the pFasL moiety. No signalling effect was detected through CD28, therefore the action of CD80 occurs via an improved targeting of the pFasL towards its molecular target.

BENEFITS The pFasL construct can be engineered to create chimeras with other cell ligands or receptors, allowing the targeting of various kinds of cell types. The generation of pFasL-derived chimeras allows to improve the efficacy of FasL apoptosis and lowering the doses to decrease the unwanted toxicity towards Fas-sensitive cells not expressing the receptor or ligand able to interact with the targeting module.

INDUSTRIAL APPLICATIONS This innovation can be used for creating highly active FasL-based apoptosis inducers with the ability to interact with molecular markers expressed by unwanted cells, e.g. tumor cells, in immunotherapy protocols.

DEVELOPMENT STAGE The cytotoxic activity of pFasL in vitro and in vivo in a mouse model of human tumor has been demonstrated (Daburon et al, PlosOne, 2013; and Morello et al, PlosOne, 2013). The enhanced activity of CD80-pFasL when compared to pFasL has been validated using Fas-positive cell lines expressing or not CD28, from T- and B-lymphocyte lineages. Experiments are underway using primary cells from CD28 and Fas positive human myeloma and plasmocytoma cells.

Reference 03946-01

Keywords IgFasL chimeric protein, Myeloma, apoptosis, Fas receptor, anti-tumoral activity. Status of Patent - Priority patent application n° EP 11 306395.2 filed on October 27, 2011 entitled "Chimeric molecule involving oligomerized FasL extracellular domain" - International patent application n°PCT / EP2012 / 071291 filed on October 26, 2012. - Validation in US & EP

Inventors Jean Luc TAUPIN Sophie DABURON Jean-François MOREAU Myriam CAPONE Commercial Status Exclusive or non-exclusive licenses, Collaborative agreement Laboratory

« Composantes innées de la réponse immunitaire et de la différenciation », a

CNRS-Ubx2 laboratory

(UMR 5164) in Bordeaux,

France. http://www.umr5164.u-bordeaux2.fr/

16 Table of content

Tumor vaccine for immunotherapy of

tumoral pathologies related to EBV

latency infections

CONTEXT The Epstein-Barr virus (EBV) is associated with various tumoral pathologies which can be distinguished by the expression of the latency antigens. Latency infections by Epstein-Barr virus remains a serious problem. The research team works on efficient immunotherapy protocols against tumoral pathologies related to EBV latency infections from type I and II.

TECHNICAL DESCRIPTION The invention relates to immunogenic peptides from Epstein-Barr Virus type I and II latency antigens, comprising at least one CD4+ T epitope which can be recognized by the majority of individuals in the Caucasian population. The inventors have identified a peptide cocktail inducing an EBV-specific CD4+ T-cell response which may be an excellent and safe candidate for a vaccine against EBV latency II malignancies.

BENEFITS This immunotherapy approch has important advantages :

No risk of aplasia as observed with radiotherapy or chimiotherapy No aggravation of the preexisting immunosuppressive environment No effect on regulatory T lymphocytes usually associated with a poor prognosis and

overall survival Cytotoxic potential even in the presence of tumoral exosomes

Limited side effects

INDUSTRIAL APPLICATIONS This innovation could be used for:

- Prevention and treatment of pathologies related to EBV latency infections, in particular Hodgkin’s lymphoma (HL), NKT Lymphoma and nasopharyngeal carcinoma (NPC).

Treatment to prevent recurrence of EBV-associated malignancies

DEVELOPMENT STAGE Accomplished tests:

- Peptides recognized by human CD4+ memory T cells from healthy donors and recognized by PBMC isolated from HL patients

- Generation of EBV-specific CD4+ T-cell lines => characterization of a specific cytotoxic activity on Hodgkin’s lymphoma and NPC cell lines

- Peptides have no effect on the phenotype and the function of human regulatory T cells

- Proof of concept with humanized SCID mice model xenotransplanted with human nasopharyngeal carcinoma and reconstituted with human PBMC => Peptides induce an inhibition of tumor growth

Reference 03946-01

Keywords Epstein Barr virus EBV; Diagnostic; Immunotherapy; Peptide vaccine; Cancer prevention and treatment ; Hodgkin’s lymphoma; Nasopharyngeal carcinoma Status of Patent

French priority patent application n° FR2881746 filed on February, 7, 2005, and entitled “ Epitopes T CD4+ des antigènes de latence de type I et II du virus Epstein-Barr aptes à être reconnus par la majorité des individus de la population caucasienne et leurs applications" Pending patent applications in CA and US. Granted patents : FR, GB, NL, CH, JP Inventors

Véronique PANCRE Claude AURIAULT Stéphane DEPIL Bernard MAILLERE Olivier MORALES Gaetan MUNIER

17 Table of content

Current developments:

- Evaluation of EBV peptides on PBMC isolated from NPC patients - Test of recognition of EBV peptides by PBMC isolated from asian donors

In vivo proof of concept in HL mouse model

Commercial Status

Exclusive or non-exclusive license Laboratory

Institut de Biologie de Lille, (IBL, UMR 8161), France http://umr8161.cnrs.fr/

Articles Mrizak et al, J Natl Cancer Inst. 2014 Dec 12;107(1):363. Morales et al., J Immunother. 2012 Apr;35(3):254-66. Depil et al., J Immunother. 2007 Feb-Mar;30(2):215-26

18 Table of content

Novel recombinant lectins and

applications for the detection of

pathological state markers

CONTEXT Changes in cell surface glycosylation are known to be associated with a large number of chronic diseases and the cancer glycans represent a very promising field for biomarker discovery. Among them, specific glycans, such as those terminated by N-acetylglucosamine (GlcNAc) residues are rare on healthy tissues. Other oligosaccharides, rich in fucose (Fuc) residues are markers of inflammation. Lectins have been demonstrated to be very efficient for detecting aberrant glycosylation in biological samples. A new technology is the use of lectin arrays that can contain lectins with large spectra of specificity and therefore characterize variation of glycosylation. The use of lectins extracted from natural organisms may be time-consuming and may generate problems of contamination or variations between batches. Therefore, recombinant lectin technology is starting to be developed.

TECHNICAL DESCRIPTION The present invention relates to recombinant multimeric lectins having a beta-propeller architecture, formed from repetition and/or oligomerisation of 4 to 7 modules of approximately 30 to 60 amino acids. The concept of neolectins consist in modifying first the valency, creating a monovalent lectin for which the specificity can be engineered, then recreating lectins with chosen number of active binding sites.

BENEFITS These neolectins have important advantages over extracted lectins. They are recombinant proteins, with higher purity (no contamination) and higher reproducibility than lectins purified from plants of invertebrates. There specificity and valency can be modified to be adapted for recognition of specific epitopes. They are easier, cheaper and faster to produce than antibodies.

INDUSTRIAL APPLICATIONS This innovation could be used for:

- Glycobiology research - Quality control of recombinant glycoproteins (for example for characterizing the

glycosylation of therapeutic antibodies) - Diagnostic / Prognostic / Therapeutic follow-up of cancers as leukaemia,

adenocarcinoma, kidney, breast and ovary carcinoma, … but also others diseases as rheumatoid arthritis, elderly dementia, …

- Diseases treatment, particularly cancers (for drug vectorisation)

Reference 06433-01

Keywords Lectins; Glycobiology; Cancer diagnostic Status of Patent Priority patent application n° FR 14 51740 filed on March 04, 2014, and entitled "Nouvelles lectines et applications pour la détection de marqueurs d'états pathologiques."

Inventors Anne IMBERTY, Julie ARNAUD, Aymeric AUDFRAY Commercial Status Exclusive or non-exclusive license Laboratory Centre de recherches sur les macromolécules végétales CERMAV (UPR5301 ), Grenoble, France. http://www.cermav.cnrs.fr/fr/glycobiologie-moleculaire

19 Table of content

DEVELOPMENT STAGE Accomplished tests:

- Production of recombinant lectin from Psathyrella velutina (GlcNAc specific) and Burkholderia ambifaria (Fuc specific)

- Production of neolectins from Ralstonia solanacearum with controlled valency - Labeling of human cancer cells and carcinoma tissues

Current developments: Modifying the specificity of lectin through phage display approach

20 Table of content

Part 1.4: Biomarkers (targets)

21 Table of content

New target involved in DNA

replication for diagnosing and

treating cancers

CONTEXT

Although DNA replication is a vital process for cell growth and its mechanism is highly conserved, recent studies also reveal significant diversity in origin structure, assembly of pre-replication complex (pre-RC) and regulation of replication initiation along evolutionary lines. It is crucial to maintain genomic integrity in eukaryotic cells. During DNA replication, errors in pre-RC assembly often result in substantial changes in the amount of genetic material, which will cause cell death, or transform cells to become malignant tumors. The first origin recognition complex (ORC) proteins were purified from cell extracts as a heterohexameric complex that specifically binds to origins of DNA replication and the subunits were named Orc1 through Orc6.

TECHNICAL DESCRIPTION The invention is based on the characterization by the inventors of a new protein that physically interacts with the ORC protein complex and that is involved in DNA replication. By a purification process of the ORC protein complex, the inventors have identified a new ORC protein complex partner which has been identified to specifically interact with both ORC1 and LRWD1 proteins. They named this protein ORC Ubiquitin-ligase 1 or Obi1. The invention relates to this protein or derived proteins as drug and to coumpounds affecting (i.e. inhibiting or activating) the expression or the activity of said protein.

INDUSTRIAL APPLICATIONS This innovation could be used for diagnosing and treating pathologies involving a deregulation of DNA replication (cancers or rare diseases as Seckel syndrome, Meier-Gorlin syndrome, Majewski osteodysplastic primordial dwarfism type II, etc.)

Reference 06081-01

Keywords DNA replication origins, ORC Ubiquitin-ligase, cancer, orphan diseases

Status of Patent Priority patent application n° EP 13305725.7 filed on May 31, 2013, entitled “Protein involved in DNA replication, and modulation of its activity.”

Inventors Marcel MECHALI, Philippe COULOMBE

Commercial Status Exclusive or non-exclusive license Laboratory Institut de Génétique Humaine (IGH), UPR1142 in Montpellier, France http://www.igh.cnrs.fr

22 Table of content

Part 2: Pronostic

23 Table of content

Development of Anti Trio antibody to

detect Trio as a potential pronostic

marker for cancer

CONTEXT Markers of prognosis are essential tools for clinicians to better predict patient treatment and represent an intense area of research especially in the cancer field. It is a great necessity to develop sensitive and specific tools for markers of prognosis. The Rho-GEF Trio is an activator of RhoGTPases. Trio has been proposed to play a role during oncogenesis, based on the observation that its expression is significantly increased in a number of cancers. For example, recent data have shown that Trio overexpression is associated to the TN-sub types of breast cancer and with sarcoma with complex genetic profile. It is a great necessity to develop specific and sensitive antibodies against Trio that can be used as potential markers of prognosis.

TECHNICAL DESCRIPTION This invention describes the design of a specific epitope in the Trio sequence and the production of a polyclonal antibody against this sequence. This antibody is specific and is more sensitive than the available commercial antibodies against Trio. This antibody can be used to test whether Trio can be a prognosis marker of different types of tumors.

DEVELOPMENT STAGE Test of antibody on Western-blot, immunofluorescence, immunohistochemistry.

BENEFITS Trio overexpression is associated to the TN-sub types of breast cancer and with sarcoma with complex genetic profile. So far, no good antibody was available for analysis by immunohistochemistry on TMA. We have developed an antibody against Trio, which is much more sensitive than the commercial antibodies.

INDUSTRIAL APPLICATIONS Applications include: potential prognosis marker for TN-sub types of breast cancer, potential prognosis for sarcoma with complex genomic profile, research tool.

Reference 05573-01

Keywords Trio, cancer, diagnostic

Status of Patent Priority patent application n° 12306543.5 filed on December the 17th , 2012 entitled " Antibody against the protein trio and its method of production "

Inventors Anne DEBANT Jérôme BOUDEAU

Commercial Status Exclusive or non-exclusive licenses, Collaborative agreement Laboratory

Centre de recherche de biochimie macromoléculaire, a

CNRS laboratory (UPR5237) in Montpellier, France. http://www.crbm.cnrs.fr/?lang=en

24 Table of content

Part 3: Theranostic

25 Table of content

A novel methodology to predict the

response to HER2 bloking agent

treatment in cancer

CONTEXT The transmembrane receptor protein HER2 has a tyrosine kinase activity and is involved in the regulation of several genes linked to the transcription of cell cycle. HER2 is overexpressed in several types of cancer, as breast, ovary, colon, pancreas…

This protein is a well known target for anticancer treatments as Trastuzumab, a monoclonal antibody, HER2 bloking agent.

HER2 positive breast cancer show different responses to a trastuzumab treatment. In several cancer cases, Trastuzumab is not indicated to treat the tumor. Indeed, medical centers try to adapt the treatment according to preliminary pronostic on the response of each patient to a HER2 blocking agent treatment.

This prognostic can be done by the characterization of each patient tumoral genes expression profile, from 4 to 28 genes on tumor biopsy. This method is long, expensive and implies a biopsy.

TECHNICAL DESCRIPTION The inventor has settled a novel methodology to predict each patient response to a HER2 blocking agent treatment. This methodology is based on a surprising discovery on connections between the negative response to Trastuzumab and the silent germinal polymorphism of a specific gene which down regulates the half-life of mRNA encoding several oncogenes, inflammatory factors and angiogenic factors. This in vitro or ex vivo method comprises 2 steps :

- The specific nucleotidic polymorphism detection on a biological sample - The determination of expression of genes of interest on a biological sample

BENEFITS This method is related to prediction of treatment efficacy which is not currently available. First of all it is very simple, fast, reliable, reproducible and relatively cheap compared to actual methods:

- No biopsy of the tumor is necessary (less risk related to this sampling). The analysis is based on a simple blood sample (5 to 10 mL) of the patient.

- Analysis involves simple and common molecular biology techniques.

INDUSTRIAL APPLICATIONS The main application of this invention is the preliminary prognostic to the response of treatment of a cancer with an HER2 blocking agent. This methodology will help medical centres to adapt HER2 positive cancer treaments.

DEVELOPMENT STAGE

Reference 04327-01

Keywords Trastuzumab, HER2,

erbB2, NEU, treatment prediction, breast cancer Status of Patent Priority patent application n° FR1155128 filed on 10/06/2011, entitled " Méthode de prédiction de la réponse à un traitement avec un agent bloquant HER2 " Inventor Gilles PAGES Commercial Status Exclusive or non-exclusive licenses, Collaborative agreement Laboratory Institut de biologie du développement et cancer (IBDC), a CNRS laboratory (UMR6543) in Nice, France. http://www.unice.fr/isdbc

26 Table of content

This methodology has been validated by inventors on a breast cancer patient cohort.

27 Table of content

Part 4: Diagnostic

28 Table of content

Use of an inhibitory protein of cell

migration as a diagnostic marker for

metastastic progression

CONTEXT Cell migration relies on the ability of the cell to project its plasma membrane. Membrane protrusion relies on the generation of a pushing force by the actin cytoskeleton. In particular the molecular machine called the Arp2/3 complex generates branched actin networks, which have been seen to develop such pushing forces. It is well stablished that the Arp2/3 complex is critical for cell migration and invasion of tumor cells through tissues.

TECHNICAL DESCRIPTION We have discovered Arpin, an inhibitory protein of the Arp2/3 complex. This protein inhibits the Arp2/3 complex at the plasma membrane, and as a consequence inhibits cell migration. Arpin impedes the efficiency of cell migration by both reducing cell speed and the directionality of cell migration (Dang et al., Nature 2013). Here we have found that Arpin expression is lost in most invasive breast carcinomas and that this loss correlates with poor prognostic for patients (Dang et al., Nature 2013).

DEVELOPMENT STAGE We are currently trying to extend the field of application of Arpin besides breast cancer. We have already identified a similar down-regulation of Arpin expression in colorectal cancers and a patient cohort is under sutdy for testing Arpin as a prognosis marker in colorectal cancer. We are also investigating the mechanisms by which tumor cells loose Arpin expression.

BENEFITS Importantly, the Arpin prognostic marker does not correlate with any other previously known marker indicating that measuring Arpin expression refines prognostic for breast cancer patients and represents a unique and unexploited marker.

INDUSTRIAL APPLICATIONS This invention should interest companies that are implicated in developing diagnostic kits for breast cancer progression. Arpin expression is a good marker both at the mRNA level and at the protein level. Thus its inclusion in any existing diagnostic chip should improve its efficiency. Further, we have demonstrated that overexpression of Arpin reduces cell migration, opening the field of peptide therapeutics (described in the patent).

Reference 06299-01

Keywords Arp2/3, cancer, diagnostic

marker, treatment metastasis. Status of Patent Priority patent application n° EP13306198 filed on September, 2013 entitled " Use of Arpin, a new inhibitor of the ARP2/3 complex for the diagnose and treatment of diseases "

Inventors Alexis Gautreau Roman Gorelik Carla Sousa-Blin Emmanuel Derivery Sophie Vacher Antonina Alexandrova Commercial Status Exclusive or non-exclusive licenses, Collaborative agreement Laboratory Laboratoire d'Enzymologie et Biochimie Structurales, CNRS UPR3082 Gif-sur-Yvette, France. http://www.lebs.cnrs-gif.fr/en/teams/gautreau.html

29 Table of content

Mannose 6-phosphate receptor,

new target for prostate cancer

diagnosis

CONTEXT Prostate cancer is the most common cancer in France and is the fourth leading cause of mortality. The health authorities highlight the need of new tissue marker identifying early forms of prostate cancer to complete the current diagnostic assays. They also stressed the importance of markers to distinguish aggressive forms from indolent forms.

TECHNICAL DESCRIPTION The overexpression of the cation-independent mannose 6-phosphate receptor (CI-M6PR also known as Insulin like growth factor 2 receptor) is demonstrated in malignant prostatic cells by immunohistochemistry on paraffin embedded tissue sections.

DEVELOPMENT STAGE The detection of CI-M6PR (IGF2R) in human prostatic tissues is conducted by a conventional immunohistochemistry method on paraffin embedded tissue biopsies. For this purpose we use our in house produced polyclonal antibody. The clinical study was realized on a cohort of 167 patients (including 41 healthy patients). 84% of the cancer positive patients were positive for CI-M6PR expression, including early cancer development stage (Gleason grade 4). No expression of CI-M6PR was observed on healthy tissues.

BENEFITS The present invention provides a specific marker for prostate cancer diagnosis Since CI-RM6P is overexpressed only in malignant part of the prostatic tissue (not in normal prostate tissue).

INDUSTRIAL APPLICATIONS Production of a new clinical kit for the early diagnosis of prostate cancer in multiple tissue biopsies.

Reference 06100-01

Keywords Prostate; cancer; diagnostic; CI-M6PR ; IGF2R Status of Patent France priority patent application n° 14 50588 filed on January 23, 2014 and entitled “Nouvelle méthode de dépistage du cancer de la prostate”

Inventors Marcel GARCIA, Ophélie VAILLANT, Xavier REBILLARD, et al. Commercial Status Exclusive or non-exclusive License Laboratory Institut des Biomolecules Max Mousseron (IBMM), UMR5247 Montpellier, FRANCE

30 Table of content

Novel recombinant lectins and

applications for the detection of

pathological state markers

CONTEXT Changes in cell surface glycosylation are known to be associated with a large number of chronic diseases and the cancer glycans represent a very promising field for biomarker discovery. Among them, specific glycans, such as those terminated by N-acetylglucosamine (GlcNAc) residues are rare on healthy tissues. Other oligosaccharides, rich in fucose (Fuc) residues are markers of inflammation. Lectins have been demonstrated to be very efficient for detecting aberrant glycosylation in biological samples. A new technology is the use of lectin arrays that can contain lectins with large spectra of specificity and therefore characterize variation of glycosylation. The use of lectins extracted from natural organisms may be time-consuming and may generate problems of contamination or variations between batches. Therefore, recombinant lectin technology is starting to be developed.

TECHNICAL DESCRIPTION The present invention relates to recombinant multimeric lectins having a beta-propeller architecture, formed from repetition and/or oligomerisation of 4 to 7 modules of approximately 30 to 60 amino acids. The concept of neolectins consist in modifying first the valency, creating a monovalent lectin for which the specificity can be engineered, then recreating lectins with chosen number of active binding sites.

BENEFITS These neolectins have important advantages over extracted lectins. They are recombinant proteins, with higher purity (no contamination) and higher reproducibility than lectins purified from plants of invertebrates. There specificity and valency can be modified to be adapted for recognition of specific epitopes. They are easier, cheaper and faster to produce than antibodies.

INDUSTRIAL APPLICATIONS This innovation could be used for:

- Glycobiology research - Quality control of recombinant glycoproteins (for example for characterizing the

glycosylation of therapeutic antibodies) - Diagnostic / Prognostic / Therapeutic follow-up of cancers as leukaemia,

adenocarcinoma, kidney, breast and ovary carcinoma, … but also others diseases as rheumatoid arthritis, elderly dementia, …

- Diseases treatment, particularly cancers (for drug vectorisation)

Reference 06433-01

Keywords Lectins; Glycobiology; Cancer diagnostic Status of Patent Priority patent application n° FR 14 51740 filed on March 04, 2014, and entitled "Nouvelles lectines et applications pour la détection de marqueurs d'états pathologiques."

Inventors Anne IMBERTY, Julie ARNAUD, Aymeric AUDFRAY Commercial Status Exclusive or non-exclusive license Laboratory Centre de recherches sur les macromolécules végétales CERMAV (UPR5301 ), Grenoble, France. http://www.cermav.cnrs.fr/fr/glycobiologie-moleculaire

31 Table of content

DEVELOPMENT STAGE Accomplished tests:

- Production of recombinant lectin from Psathyrella velutina (GlcNAc specific) and Burkholderia ambifaria (Fuc specific)

- Production of neolectins from Ralstonia solanacearum with controlled valency - Labeling of human cancer cells and carcinoma tissues

Current developments: Modifying the specificity of lectin through phage display approach

32 Table of content

CNRS Key figures Source: CNRS internet website

Budget forecast for 2014

3.3 billion Euros

Personnel

Over 32,000 employees of which 24,955 are CNRS tenured employees: o 11,204 researchers o 13,751 engineers and support staff

Organization

10 institutes (3 of which have the status of national institutes) 19 regional offices, ensuring decentralized direct management of laboratories 1,028 research units (95 % are joint research laboratories with universities and industry)

International relations

50 exchange agreements (with over 40 countries) 4,600 foreign visiting scientists (PhD students, post-docs and visiting researchers) 1,690 permanent foreign researchers 420 permanent foreign engineers and technicians 318 International Programs for Scientific Cooperation (PICS) 161 International Associated Laboratories (LEA/LIA) 103 International Research Groups (GDRE/GDRI) 26 International Joint Units (UMI) 8 CNRS offices abroad (Beijing, Brussels, New Dehli, Pretoria, Rio de Janeiro, Tokyo,

Washington, Singapur)

Industrial relations

25 framework agreements with major international industrial groups 4,521 main patents (is in the Top 100 Global Innovators, and 7th applicant in France) 959 licenses and other financially remunerating active acts 704 companies created with CNRS since 2000

Awards

19 Nobel Prizes 12 Fields Medals

33 Table of content

www.fist.fr