COMMITTEE FOR ORPHAN MEDICINAL PRODUCTS

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1 European Medicines Agency Pre-authorisation Evaluation of Medicines for Human Use COMMITTEE FOR ORPHAN MEDICINAL PRODUCTS Document Date: London, 10 July 2008 Doc.Ref.: EMEA/COMP/130384/2008 PUBLIC SUMMARY OF POSITIVE OPINION FOR ORPHAN DESIGNATION OF recombinant human histone and recombinant human N-bis-met-histone for the treatment of acute myeloid leukaemia On 20 December 2007, orphan designation (EU/3/07/516) was granted by the European Commission to SymbioTec GmbH, Germany, for recombinant human histone and recombinant human N-bismet-histone for the treatment of acute myeloid leukaemia. What is acute myeloid leukaemia? Acute myeloid leukaemia is a disease in which cancer cells are found in the blood and the bone marrow. The bone marrow is the spongy tissue inside the large bones in the body. Normally, the bone marrow makes cells called blasts, which mature into several different types of blood cells that have specific functions in the body. These include red cells, white cells and platelets. Red blood cells carry oxygen and other materials to all tissues of the body. White blood cells fight infection. Platelets make the blood clot. When leukaemia develops, the bone marrow produces large numbers of abnormal blood cells. There are several types of leukaemias. In myeloid leukaemia, blasts that should develop into a type of white blood cells called granulocytes are affected. The blasts do not mature, and become too many. These blast cells are then found in the blood; they also accumulate in the bone marrow where they take the place of the other types of normal blood cells, causing anaemia, easy bruising, and frequent infections. Myeloid leukaemia can be acute, when it develops quickly with many blasts. Acute myeloid leukaemia is life-threatening. What are the methods of treatment? Treatment for leukaemia is complex, and depends on a number of factors including the type of leukaemia, the extent of the disease and whether the leukaemia has been treated before. It also depends on the age, the symptoms, and the general health of the patient. The primary treatment of acute myeloid leukaemia is chemotherapy (using drugs to kill cancer cells). Several products were authorised for the condition in the community at the time of submission of the application for orphan drug designation. Satisfactory argumentation has been submitted by the sponsor to justify the assumption that recombinant human histone and recombinant human N-bis-met-histone might be of potential significant benefit for the treatment of acute myeloid leukaemia because of its mechanism of action which is different from existing treatments. This assumption will have to be confirmed at the time of marketing authorisation. This will be necessary to maintain the orphan status. What is the estimated number of patients affected by the condition *? Based on the information provided by the sponsor and previous knowledge of the Committee, acute myeloid leukaemia was considered to affect less than approximately 2 in 10,000 persons in the European Union, which, at the time of designation, corresponded to about 100,000 persons. * Disclaimer: For the purpose of the designation, the number of patients affected by the condition is estimated and assessed based on data from the European Union (EU 27), Norway, Iceland and Lichtenstein. This represents a population of 498,000,000 (Eurostat 2006). This estimate is based on available information and calculations presented by the sponsor at the time of the application. 7 Westferry Circus, Canary Wharf, London, E14 4HB, UK Tel. (44-20) Fax (44-20) [email protected] European Medicines Agency, Reproduction is authorised provided the source is acknowledged

2 How is this medicinal product expected to act? Histones including histone are naturally occurring proteins of the human body where they predominantly occur in the nucleus of cells. Recombinant N-bis-met-histone is an artificial molecule (a prodrug) which is transformed in the body to histone. Tumour cells differ from healthy cells as their outer membrane bear receptors (proteins that bind specific molecules) which bind histone and N-bis-met-histone. These histones and the receptors form large aggregates in the membrane, leading to membrane ruptures and the ultimate death of the cancer cells. What is the stage of development of this medicinal product? The effects of recombinant human histone and recombinant human N-bis-met-histone were evaluated in experimental models. At the time of submission of the application for orphan designation, clinical trials in patients with acute myeloid leukaemia were ongoing. Recombinant human histone and recombinant human N-bis-met-histone was not authorised anywhere worldwide for the treatment of acute myeloid leukaemia, at the time of submission. Orphan designation of amonafide l-malate was granted in the United States for the treatment of acute myeloid leukaemia. According to Regulation (EC) No 141/2000 of 16 December 1999, the Committee for Orphan Medicinal Products (COMP) adopted on 8 November 2007 a positive opinion recommending the grant of the above-mentioned designation. Opinions on orphan medicinal products designations are based on the following cumulative criteria: (i) the seriousness of the condition, (ii) the existence or not of alternative methods of diagnosis, prevention or treatment and (iii) either the rarity of the condition (considered to affect not more than five in ten thousand persons in the Community) or the insufficient return of development investments. Designated orphan medicinal products are still investigational products which were considered for designation on the basis of potential activity. An orphan designation is not a marketing authorisation. As a consequence, demonstration of the quality, safety and efficacy will be necessary before this product can be granted a marketing authorisation. For more information: Sponsor s contact details: SymbioTec GmbH Science Park Saar 1 D Saarbruecken Germany Telephone: Telefax: [email protected] EMEA 2008 Page 2/5

3 Patients associations contact points: Cancer BACUP 3 Bath Place Rivington Street London EC2A 3JR United Kingdom Telephone: Telefax: / (freephone for UK) Ligue Nationale contre le Cancer (LNCC) 12, rue Corvisart Paris France Telephone : Telefax : [email protected] EMEA 2008 Page 3/5

4 Translations of the active ingredient and indication in all EU languages and Norwegian and Icelandic Language Active Ingredient Indication English Recombinant human histone and recombinant human N-bis-met-histone Bulgarian Czech Danish Dutch Estonian Finnish Рекомбинантен човешки гистон и рекомбинантен човешки Н-бис-метгистон Rekombinantní lidský histon a rekombinantní lidský N-bis-met-histon Rekombinant humant histon og rekombinant humant N-bis-met-histon Recombinant humaan histon en recombinant humaan N-bis-met-histon Rekombinantne inimese histoon ja rekombinantne inimese N-bis-met-histoon Rekombinantti ihmis-histoni ja rekombinantti ihmis-n-bis-met-histoni French Histone humaine recombinante et N- bis-met-histone humaine recombinante German Rekombinantes humanes Histon und rekombinantes humanes N-bis-met-Histon H1. Greek ανασυνδυασμένη ανθρώπινη ιστόνη και ανασυνδυασμένη ανθρώπινη N-bismet-ιστόνη Hungarian Italian Latvian Lithuanian Rekombináns humán hiszton és rekombináns human N-bis-met-hiszton Istone umano ricombinante e N-bismet-istone umano ricombinante Rekombinēts cilvēka histons un rekombinēts cilvēka N-bis-met-histons Rekombinantinis žmogaus histonas ir rekombinantinis žmogaus N-bis-methistonas Maltese Istone uman rikombinanti tat-tip u N- bis-met-istone uman rikombinanti tat-tip Treatment of acute myeloid leukaemia Лечение на остра миелоидна левкемия Léčba akutní myeloidní leukémie Behandling af akut myeloid leukæmi Behandeling van acute myeloïde leukemie Akuutse müeloidse leukeemia ravi Akuutin myelooisen leukemian hoito Traitement de la leucémie aiguë myéloïde Behandlung der akuten myeloischen Leukämie Θεραπεία της οξείας μυελοειδούς λευχαιμίας Akut myeloid leukaemia kezelése Trattamento della leucemia mieloide acuta Akūtas mieloleikozes ārstēšana Ūmios mieloleukozės gydymas Kura tal-lewkimja mjelojda akuta Polish Ludzki histon rekombinowany oraz ludzki N-bis-met-histon rekombinowany Leczenie ostrej białaczki szpikowej EMEA 2008 Page 4/5

5 Portuguese Histona recombinante humana e N- bis-met-histona recombinante humana tipo Romanian Histonă umană recombinantă şi N- bis-met-histonă umană recombinantă Slovak Rekombinantný ľudský histón a rekombinantný ľudský N-bis-met-histón Slovenian Ljudski rekombinantni histon in ljudski rekombinantni N-bis-met-histon Spanish Histona recombinante humana y N- bis-met-histona recombinante humana Swedish Rekombinant humant histon och rekombinant humant N-bis-met-histon Norwegian Rekombinant humant Histon og rekombinant humant N-bis-met-histon Icelandic Raðbrigða manna histón og raðbrigða manna N-bis-met-histón Tratamento da leucemia mielóide aguda Tratamentul leucemiei mieloide acute Liečba akútnej myeloickej leukémie Zdravljenje akutne mieloične levkemije Tratamiento de la leucemia mieloide aguda Behandling av akut myeloisk leukemi Behandling av akutt myelogen leukemi Meðferð við bráðu kyrningahvítblæði EMEA 2008 Page 5/5

6 Non-Confidential Factsheet ONCOHIST - a paradigm shift in cancer therapy - A novel product offering for licensing Developed by: Represented through: SYMBIOTEC GmbH Tytonis BV Science Park Saar 1 Van der Kaaijstraat 64 D Saarbrücken 1815 VM Alkmaar Germany The Netherlands For further information, please contact: Michael Zeppezauer, Prof. Dr., , or [email protected] ( ) Henrik L. Luessen, Ph.D., (cell phone), or [email protected] ( ) OncoHist Non-Confidential Factsheet 1/14

7 Executive Summary The development of SymbioTec s medicament is based on 25 years research concerned with novel functions of histones in the group of prof. Michael Zeppezauer (co-founder of SymbioTec) in the University of the Saarland. The most important published papers are: (Reichhart et al, 1985) and (Class et al, 1996) and numerous patents. Histone H1 is a highly conserved protein with strong anti-proliferative properties against cancer cells of different histological origin. This has been demonstrated extensively for hematologic malignancies, such as leukemias, lymphomas, and myelomas (Class, Lindman et al. 1996), but also for tumors from other tissues. Susceptibility of cells to the cytotoxic effect of histones is determined by the ability of histone H1 to destabilize the tumor cell membrane, which results in cell lysis and death. Phosphatidylserine-exposure in endothelial cells of tumor vasculature suggests potential anti-angiogenetic activity of histone H1 (patent pending). This would result in a dual way of attacking solid tumors by histone H1: oncolysis and angiogenesis-inhibition. Histone therapy is defect-oriented against pathogenic cells and it has clear advantages vis-a-vis current function-oriented, targeted therapies. Delivery carriers for histone drugs are not necessary. Histone H1 can cross the blood-brain barrier; therefore it might be useful in treating distant brain metastases and as transport vehicle for other drug molecules (Pardridge, Triguero et al. 1989). Since histone H1 is an evolutionary extremely well conserved protein with very limited interspecies variation, it possesses low immunogenicity and antigenicity, making the stimulation of neutralizing antibodies or therapy-limiting immune responses in recipients unlikely. In 1984 the first patent application was filed for the general use of histones in diagnostic and therapeutic procedures, in 1989 an application was filed for the dedicated use of histones H1 and H2A:H2B in cancer therapy. Since then about 40 patents were granted or filed in 10 patents families, protecting the use of histones in different fields, e.g.: Cancer and side effects of cancer: - As mono- and combination-therapy - Thrombocytopenia - Tumor angiogenesis Other diseases: - Viral infections - Bacterial infections - Autoimmune diseases OncoHist Non-Confidential Factsheet 2/14

8 Tumor therapy with natural histones is protected until In April 2007 a new patent application was filed in the European Patent Office for the protection of the new chemical entity, N-bis-methistone in use against cancer, providing additional protection until (PCT/EP2008/002746, In 2001 SymbioTec GmbH has achieved its first financing round with venture capital and funds from private investors and started to work in new laboratories in Science Park Saar 1. The goal of the company was to develop expression systems for recombinant human histone, a manufacturing process for the active substance and to develop the analytical methods to analyse the drug identity, purity and activity. These goals were achieved. The methods used were validated. Proof of activity on leukemic cell lines was validated with the recombinantly manufactured drug substance. The production and purification methods were transferred to a Contract Manufacture Organisation (Eurogentec). Drug products (GLP) were tested toxicologically by the renowned Dutch CRO NOTOX B.V. in 2003 and between 2004 and 2008 the first clinical phase I/II trial with AML patients was successfully conducted. This is the very first clinical trial in which histone proteins have been given intravenously to humans Orphan Drug Status (Orphan Medicinal Product Designation) was awarded to SymbioTec for the active ingredients of ONCOHIST for treatment of AML by the European Commission in December 2007 and by the FDA in October The compound was tested in almost 60 human cancer cell lines from different tissues and showed high cytotoxic efficacy throughout, which would suggest the potential to have a universally acting oncolytic compound at hand that could also reach metastatic cancer. The fact that in the recent clinical trial no MTD was reached until the dose of 628 mg/m 2 body surface area would also make the drug compound interesting for combination therapies. SymbioTec is planning the perfomance of a complete clinical phase II trial in the indication acute leukemias (AML and ALL) with the prospect of a fast track approval after completion of this trial. The first phase II clinical trial with 120 AML patients has recently been started in three renowned clinical centers in the Russian Federation: Federal Hematology Scientific Center (Moscow), Federal Hematology and Transfusion Research Center (St. Petersburg) and St. Petersburg Medical Academy (St. Petersburg).. The aim of this trial is to examine the benefits of the combination therapy of ONCOHIST with two regimens of standard therapy: HAM (cytarabine with mitoxantrone) and low-dose-cytarabine (LDAC). OncoHist Non-Confidential Factsheet 3/14

9 The drug compound Histone H1 is one of the most abundant proteins in the nucleus. Remarkably, despite years of study, the most basic questions concerning H1, such as its precise locations in chromatin and many aspects of its function, remain controversial or unanswered. Histone H1 has long been known as the linker histone, on the basis of classic studies that showed H1 to be somehow located on, or to contact, the linker DNA that goes between the nucleosomes that are the basic structural units of chromatin. Historically, the support of structural integrity of chromatin was the only recognized function of the histone H1 family. Nowadays the active involvement of the histone proteins in the phenomenology of gene regulation and transcription is widely recognized under the term epigenetics. However, the systemic biology of the histone protein family is much more diverse and displays a fascinating multifunctionality outside the cell nucleus and outside the cells: histone proteins and histone-derived peptides are regulatory components of the immune system with anti-inflammatory activity and act as strong antimicrobial agents in the body fluids (e.g. amnion), in the skin and in the small intestine; thus complementing their oncolytic activity as components of the innate immune system. ONCOHIST (recombinant human ) is a small protein (molecular weight 22.2 kda) consisting of three discernible domains: A central globular domain (65 amino acids), flanked by an NH2- terminal of 40 amino acids and a COOH-terminal tail of 111 amino acids: MMSETAPLAPTIPAPAEKTPVKKKAKKAGATAGKRKA SGPPVSELITKAVAASKERSGVSLAALKKALAAAGYD VEKNNSRIKLGLKSLVSKGTLVQTKGTGASGSFKLNK KAASGEGKPKAKKAGAAKPRKPAGAAKKPKKVAG AATPKKSIKKTPKKVKKPATAAGTKKVAKSAKKVKTP QPKKAAKSPAKAKAPKPKAAKPKSGKPKVTKAKK AAPKKK OncoHist Non-Confidential Factsheet 4/14

10 In physiological buffers (e.g. saline), the N- and C-terminal tails are random coiled, whereas the central domain contains three α-helices, two amphipathic and one hydrophobic helix, and an additional β-structure. The histone H1 proteins are highly soluble in water (up to 100 mg/ml). ONCOHIST consists of mature recombinant human histone and its artificial precursor molecule N-bis-methionine-histone. and is supplied at 10 and 20 mg/ml in 0.9% NaCl or phosphate buffered saline as a colorless, sterile, and preservative-free solution in single-use glass vials with a rubber septum (3.5ml). Mode of action Phenomenologically, the rapid binding of histone H1 proteins on the surface of cancer cells is followed by apoptosis in the time scale of minutes and necrosis in the time scale of hours. Concerning the molecular level (picture) two types of recipient (receptor) molecules have been identified in the outer membrane: (i) core histones / nuclesosomes and (ii) misplaced phosphatidylserine due to the breakdown of the control of the lipid distribution in the membrane of cancer cells. (i) In several types of stressed cells e.g. activated peripheral blood lymphocytes and many cancer cells, nucleosomes appear on the outer cell membrane (Watson et al, 1995). These misplaced core histones were identified as receptors for externally added histone H1 by SymbioTec s scientists. SymbioTec s European Patent (expiry date 2018) protects the utilization of histones or their therapeutically effective parts for producing a medicament for the therapy of cancer diseases which are characterized by the fact that their cancer cells contain receptors in their membranes which consist of the histones H2A, H2B, H3 and/or H4. Especially histone H1 molecules, appropriately termed linker histones can link together several nucleosomes to larger aggregates. In the outer membranes of cancer cells these superaggregates cause ruptures in the membrane and the death of the cancer cell as shown below. OncoHist Non-Confidential Factsheet 5/14

11 (ii) In healthy cells, negatively charged phospholipids, like phosphatidylserine molecules are building blocks of the inner layer of the cell membrane. In stressed cells, like cancer cells, the PS molecules escape the cell s control and become erroneously dislocated on the outside of the membrane. Here they act as receptor molecules for histone H1, forming superaggregates which cause major rearrangements in the membrane, pore formation and ultimately the death of the cell as shown by the group of Kinnunen (Zhao et al, 2004). It is noteworthy that the endothelial cells in blood vessels inside solid tumors suffer from the same stress situation as cancer cells and thus are also amenable to attack by externally supplied histone H1 (see below). These two different kinds of receptors are also recognised by the active substances of ONCOHIST, followed by binding, formation of superaggregates, pore formation and cell lysis. The simplified mechanism is shown in the picture above using PS (phosphatidylserine) as an example. We could demonstrate by confocal microscopy that leukemia cells bind recombinant human histone (rh), labeled with the fluorescent dye Alexa Fluor 488. After a short while these molecules form patches on the surface, indicating oligomerization of the protein within the membrane of the tumor cell). OncoHist Non-Confidential Factsheet 6/14

12 Snapshot of leukemia cells (U973) during apoptosis stained with histone H1 labeled with Alexa Fluor 488 (green) and counterstained with DAPI (in red, pseudo-color) to visualize the nucleus. Implications of this mechanism of action: - Defect-oriented, not function-oriented, therefore no serious side effects to be expected (clinically proven) - The human body s own drug, no xenobiotic, therefore no resistance to be expected (experimentally proven) - Conservative protein, no immunogenicity (clinically proven) - Membrane defects universal in cancer cells of different histological origin (application possible not only for hematopoietic cancers but also for solid tumors) - Membrane defect in endothelial cells of tumor blood vessels basis for anti-angiogenetic effect of histones in solid tumors Cell Studies The cytotoxic effect of histone H1 on tumor cells was tested on 58 tumor cell lines derived from various tissues, shown in the table below. Hematopoietic tumor cell lines (indicated in blue) were among the most sensitive cell lines. The majority of cell lines were completely eradicated at histone H1 concentrations between 3 and 5 μm, the breast cancer cell line MCF-7 at about 10 µm. Recent data of two non-small cell lung cancer cell lines show IC50 values at about 5-7 µm. OncoHist Non-Confidential Factsheet 7/14

13 Cell Line Type Cell Line Type KG-1 Acute myelogenous leukemia MDA-MB-231 Breast Carcinoma HEL Acute myeloid leukemia MDA-MB-468 Breast Carcinoma THP-1 Acute myeloid leukemia T57 Cl 1 Breast Carcinoma HL-60 Acute promyelocytic leukemia MG Carcinoma X63-AG8-653 B-cell leukemia SW 1116 Colorectal carcinoma Daudi B-cell lymphoma SW 837 Colorectal carcinoma EB-2 B-cell lymphoma SW707 Colorectal carcinoma EB-3 B-cell lymphoma WC 008 Colorectal carcinoma Namalwa B-cell lymphoma WC 010 Colorectal carcinoma SZ-4 Cutaneous T-cell leukemia A431 Epidermoid carcinoma U937 Histiocytic lymphoma A1207 Glioblastoma RBL-1 Leukemia A1235 Glioblastoma 855 Lymphocytic leukemia F39 Glioblastoma CCRF-CEM Lymphocytic leukemia U138-MG Glioblastoma CCRF-SB Lymphocytic leukemia U373MG Glioblastoma CLL 547 Lymphocytic leukemia U87-MG Glioblastoma IM-9 Lymphoid leukemia EG Melanoma OH77 Lymphoma HP Melanoma HMC-1 Mastocytic leukemia RPMI 1846 Melanoma MB-02 Megacaryocytic leukemia SB Melanoma MBL-4 Megacaryocytic leukemia BAW Meningosarcoma K562 Monocytic leukemia CHO Ovarian carcinoma P388D1 Monocytic leukemia PC-3 Prostate carcinoma WEHI-3 Monocytic leukemia 4197 Squamous cell carcinoma HUT 78 T-cell leukemia HeLa Squamous cell carcinoma Jurkat T-cell leukemia Molt 3 T-cell leukemia Molt 4 T-cell leukemia Raji T-cell leukemia GH Adenocarcinoma EBV-B 010 B-cell leukemia BT-20 Breast Carcinoma MCF-7 Breast Carcinoma A drug intended for therapeutic use should be very efficient in killing cancer cells at low concentrations and have limited impact on normal cells. The following experiments were conducted to compare tissue-derived H1 and chemotherapeutic agents for their effects on leukemia cells and hematopoietic stem cells. 80 H1 5-FU The viability of leukemia cells was evaluated in a 60 BCNU direct comparison of rh and 5-fluorouracil 40 Ara-C 20 VP-16 (5-FU), cytarabine (Ara-C), carmustine (BCNU), 0 and etoposide (VP-16). As shown in the picture -20 to the left, rh clearly killed more leukemia cells than all of the chemotherapeutic drugs Drug Concentration [µm] tested. Cytotoxicity Index [%] OncoHist Non-Confidential Factsheet 8/14

14 To compare the effects of histone H1 and 1000 chemotherapeutic drugs on normal cells, 100 Histon H1 stem cells were purified from umbilical cord 10 Mtx VP-16 blood and grown in cytokine-supplemented 5-FU Paraplatinum growth media. The chemotherapeutic drugs 1 Taxol Methotrexate (Mtx), Etoposide (VP-16, 5- Fluorouracil (5-FU), Paraplatinum, and Drug Concentration [µm] Taxol were tested on the same stem-cell sample with tissue-derived H1 to avoid donor-dependent variations. Proliferation and differentiation were monitored over time. As shown in the left picture, fractional survival of stem cells was much higher for H1 than for the chemotherapeutic agents. Fractional Survival [%] Preclinical Studies Toxicology A full preclinical toxicity programme was performed by the GLP-certified professional contract research organization NOTOX B.V. As a result of the subchronic multiple dose toxicity study no animal showed any lasting damage that could be attributed to administration of rh. Clinical biochemistry and hematology profiles remained normal. Post-mortem necroscopy and pathology did not show any significant treatment-related effects. The serum half life in a 5 days intravenous toxicokinetic study was determined to be between 29 and 40 hrs for doses of 10 and 60 mg/kg body weight, respectively. In-vivo oncolytic effects The cytotoxic effects of H1 were evaluated in an animal tumor model. Daudi cells (human B-cell leukemia) were injected subcutaneously (1x10 7 per animal). Animals were observed daily. Subsequent tumor volume was determined every other day by measuring length, width and depth of the tumor using calipers. As soon as the tumor reached a volume of >300 mm 3, 50µl H1 at 5 µg/ml in PBS (i.e. 250 µg) were injected into the tumor. Control animals received PBS alone. OncoHist Non-Confidential Factsheet 9/14

15 As shown in the left picture, a single injection of H1 250 µg (indicated by the arrow) to the site of the tumor arrested the growth of the subcutaneous human B-cell lymphoma in nude mice Tumor Volume [mm3] Time [days] C1 C2 #1 #2 #3 whereas the tumor in PBS-treated mice continued to grow. At the end of the experiment, all animals were euthanized, and necropsy was performed. In the mice treated with H1, the remaining tumor appeared necrotic. With respect to the fact that most tumors had reached a volume between 500 and 600 mm 3, this experiment indicates strongly the anti-angiogenetic function of histone H1. Clinical studies After successful completion of the toxicology studies a pilot clinical trial, phase I/II with a classical design with 22 AML patients has been conducted at the Saarland University Hospital. 17 patients ccompleted treatment per protocol. The primary objective was the evaluation of safety and tolerability. For ethical reasons, only patients were enrolled who had undergone all other possible therapies and had a life expectancy of not less than one month. Despite this fact, first signs of clinical efficacy were observed: decrease of blasts in the bone marrow in three patients and improvement of the blood parameters in seven patients (examples see below). Examples of thrombocyte counts increase: Thrombocytes : start - best value Patient No. Dose level [x 10 9 /l] Time of best value follo w up 3 after cycle follow up follow up ,6, follow up 2 after cycle 2 Normal value: x 10 9 /l 1 and 16 months after follow up 3 life-threatening OncoHist Non-Confidential Factsheet 10/14

16 Examples of decrease of blasts in bone marrow: Patient No. Dose level Bone marrow blasts - screening [%] Best value [%] Time of best value * 20 cycle 2, day * 10 cycle 1, day ,6,7 80* 25 cycle 2, day 29 * Bone m arrow crumble 2 Dose level 2: 60 mg/m Dose level 6: 392 mg/m 2 Dose level 7: 628 mg/m 2 Examples of leukocyte counts increase: Patient No. Dose level Leukocytes count from start - 9 best value [x 10 /l] Time of best value day day follow up day ,6, follow up 3 Normal value: x 10 9 /l Life-threatening Most notably, the two patients (13 and 27) who had received two treatment cycles, experienced a remarkable stabilization of 17 and 7 months, respectively. The tolerability and safety were excellent, in contrast to chemotherapeutics. ONCOHIST also proved to be immunologically safe. AAI Pharma has been supporting SymbioTec as Contract Research Organization in this study. The final Clinical Study Report of AAI Pharma from is stating inter alia that: No serious side-effects were observed except for one atrial fibrillation under infusion of rh, which was considered to be possibly related to the study drug. Seventeen (17) patients completed one course of therapy (8-9 administrations), and two responding patients received a second course without side effects. No dose-limiting toxicities were observed and the maximal tolerated dose has not been reached at 628 mg/m 2. None of the treated patients developed auto-antibodies against the proves that the study drug was immunologically safe. study drug, which OncoHist Non-Confidential Factsheet 11/14

17 The most important result from the pilot clinical trial is the finding that ONCOHIST is tolerated well by patients, i.e. without side effects, contrary to chemotherapeutics. This result is in good accordance with our preclinical studies, showing that the recombinant human histone as well as the bis-met-histone derivative do not damage healthy blood cells and do not cause the development of resistant cancer cells. Despite the fact that an optimal treatment schedule has not yet been established, first clinical effects have been already observed in 7 from 17 patients, most of them already at low dose levels. This implies that an optimal treatment schedule will most probably be found using dose levels well below the maximal tolerated dose. The clinical trial had to be interrupted due to exhausted funds. Future development Clinical Phase II trial: SymbioTec is now in a fund - raising process. The aim of this financing round of SymbioTec is the perfomance of a complete clinical phase II trial in the indication acute leukemia (AML and ALL) with the aim of establishing optimized treatment schedules and dose levels for proof of efficacy. The future development depends on the funding sum obtained and on the decisions of the future investors. Alternatively, the company is prepared to out-license the product at this current state to a reputable pharmaceutical company capable to commercially develop the product. Combination therapy: In October 2009 a phase II clinical trial with 120 AML patients has been started in three renowned clinical centers in the Russian Federation: Federal Hematology Scientific Center (Moscow), Federal Hematology and Transfusion Research Center (St. Petersburg) and St. Petersburg Medical Academy (St. Petersburg). The aim of this trial is to examine the benefits of the combination therapy of ONCOHIST with two regimens of standard therapy: HAM (cytarabine with mitoxantrone) and low-dose-cytarabine (LDAC). Orphan Drug Status: Orphan Medicinal Product Designation (OMPD) implies for a medicament, inter alia, the chance of accelerated market authorization already after a successful clinical phase II trial ( fast track approval ). The active ingredients of ONCOHIST, recombinant human histone and recombinant human N-bis-met-histone have been awarded by the European Commission the Orphan Medicinal Product Designation for the treatment of Acute Myeloid Leukemia on (EU orphan designation number: EU/3/07/516). In October 2008 the OMPD for the active ingredients of ONCOHIST for the treatment of Acute Myeloid Leukemia was granted by the FDA. OncoHist Non-Confidential Factsheet 12/14

18 At present SymbioTec is preparing applications to EMEA and to FDA for Orphan Medicinal Product Designations for the active ingredients of ONCOHIST for the treatment of acute lymphoblastic leukemia ALL. With Orphan Drug Status, after the completed phase IIb, the fast tract approval (USA) and conditional marketing authorisation (EU) is a realistic chance for ONCOHIST to gain the market entry. Therapeutic perspectives Solid tumors: The blood cancer AML was chosen as the first indication to be tested in a clinical trial because of the chance to obtain a fast track approval already after a successful phase II clinical trial in accordance with the Orphan Drug Status. However, our broad screening of cell lines (see above) revealed in addition to 25 histone sensitive leukemic and lymphoma cell lines also 18 carcinoma cell lines derived from many different tissues. SymbioTec s strategic intention has been to pursue its drug development for these dominant fractions of malignant diseases after the fast-track market entry. The feasibility of this approach has already been documented by Peregrine Pharmaceuticals ongoing clinical trials using anti-phosphatidylserine antibodies (Bavituximab) targeting the membrane defects of endothelial cells in tumor blood vessels. Combination therapy: The favorable effect of ONCOHIST on the hematopoietic system and the lack of side effects (see above) render it very attractive also for combination therapy with cytostatic drugs or radiation. Aftercare: In addition, even after successful chemo- or radiation therapy of different malignancies many patients suffer from thrombocytopenia and/or neutrocytopenia as long-term side effects of these therapies. Also these patients represent a huge market potential for a post-treatment. Antibiotic potential: The remarkable antibiotic potential of histone H1 can only be mentioned in this context: particularly fascinating is the physiological role of histone H1 in the small intestine. For decades the medical community has tried to find a plausible explanation for the scarcity of tumors in the small intestine, where only 2% of all gastrointestinal tumors are found (Tierney et al, 2007). Rose et al. (Rose et al, 1998) could prove convincingly that the epithelial cells of the ileum contain considerable amounts of histone H1 in their cytosol which is released into the lumen upon apoptotic detachment of the cells. This released histone H1 is not only responsible for the maintenance of a low concentration of bacteria in this compartment (Rose et al, 1998), but obviously also for the lower incidence of malignant processes due to its selective cytotoxic activity against tumor cells. OncoHist Non-Confidential Factsheet 13/14

19 Concluding statement The ONCOHIST development has proven to achieve all milestones planned according to budget and timelines as anticipated by Symbiotec in the past. The product appears attractive, since Effectiveness of the drug has been confirmed in cell models and animal models No toxicity has been observed (both in animals and humans a MTD could not be reached at 628 mg/m 2 ) No immunogenic properties have been found either in animals and humans so far The drug components can be made recombinantly and the process has been established and validated A product formulation is available and can be made according to GMP The clinical trial reveals that the drug is well tolerated and shows improvement of the disease in some AML patients that were not responsive to other treatments any longer 40 granted patents in 10 patent families providing protecting until at least 2017 A patent application on bis-met-histones as a family of new chemical entities has been submitted providing further protection until 2027 The ONCOHIST has the potential for use in a large variety of cancer diseases and their aftercare. ONCOHIST bears a considerable, unexplored potential in the fight against infection diseases caused by multiresistent bacteria and by pathogenic fungi. ONCOHIST therapy is addressing unmet medical needs in large markets beyond AML! References: A full reference list will be provided upon request. A full and much more extensive set of data will be available upon signing a non-disclosure agreement. OncoHist Non-Confidential Factsheet 14/14