Company

Immunotech S. A. (IMT) is a private biotech company with headquarters in Buenos Aires, Argentina. It is focused on discovering and developing innovative pharmaceutical products to accelerate organ and tissue repair and to prevent infectious diseases. Currently, IMT efforts are based on proprietary molecules (synthetic oligonucleotides) with bio-regulatory properties.

IMT staff members include biologists, biochemists, physicians, pharmacists and financial experts. They are highly motivated to aggressively drive IMT candidate products through clinical trials, in order to make them available to patients worldwide.

IMT adheres to GCPs and maintains a high level of quality standards regarding data management.

Technology
IMT oligonucleotides (IMT ODNs) are approximately 24 nucleotides long and contain the sequence PyNTTTTGT, wherein Py is C or T and N is any deoxynucleotide, as the active portion of the molecule. IMT studies have demonstrated that these ODNs, mainly in their phosphorothioate version, can interact with certain human cells modifying their behaviour. Primary target cells of IMT ODNs include mesenchymal stem cells precursor cells (MSCPC), B lymphocytes, plasmacytoid dendritic cells and natural killer cells (NK cells). Upon interaction with IMT ODNs these cells can release cytokines and express cell surface proteins which orchestrate the response of the body to damage caused by foreign agents such as invading micro-organisms or trauma. In POC trials, IMT ODNs have been demonstrated to be useful in a wide field of medical applications especially as regenerative-agents in tissue repair medicine.

IMT ODNs and Tissue Repair by Adult Stem Cells

Stem cells (SC) are cells that have the ability to continuously divide and differentiate into various kinds of cells which take part in the formation of tissues and organs (e.g. bone cells, hepatocytes, neurons etc). Embryonic stem cells (ESC) are derived from the inner cell mass of the blastocyst embryonic stage. They can be maintained in culture for an extended period of time without losing differentiation potential. The successful isolation of human ESC has recently raised the hope that these cells may provide a universal tissue source to treat many human diseases. However, there are objections to the use of ESC, from both the medical and from the moral point of view. Moral issues are mainly related to the fact that the only way to obtain these cells is to destroy a fiveday-old living human embryo. Medical issues are related to the possibility of introducing with the transplanted cells infection or genetic disorders including cancer. Other stem cell populations are present in most adult tissues, and in general, their differentiation potential is restricted to the local cell diversity. These cells are reservoirs of reparative cells that are rapidly in action following injury. They migrate to the injured site where, in cooperation with other cells, participate in the repair process. Mesenchymal stem cells (MSCs) are cells found most commonly in bone marrow, blood, dermis and periosteum that are capable of differentiating into bone, cartilage, fat, tendon, muscle, hepatic, renal, cardiac, and neural cells depending upon influences from various bioactive factors.

MSC cultured in vitro have the ability to differentiate into different cell specific lineages when introduced in vivo at the site of a given damaged tissue. Therefore, MSC are a potentially valuable material for medical applications such as systemic transplantation for systemic diseases, local implantation for local tissue defects, gene therapy protocols or generation of transplantable tissues and organs in tissue engineering protocols. Harvesting MSCs from any of their sources requires minimal invasive procedures, but the number of MSCs that can be obtained from a single donor is limited. Therefore, amplification procedures are needed for medical applications to succeed.

IMT ODNs augment the number of Bone Marrow-derived MSC colonies obtained in vitro

MSC progenitor cells present in bone marrow aspirates can be cultured in vitro by using appropriate media. If IMT ODNs are added to the media, the number of MSC colonies recovered from the culture at a given time can be greatly augmented (Fig. 10).

30 Number of CFU-F / 2x10 6 cells 25 20 15 10 5 0 C o n tr o l IM T 5 04

Stem cells (2007);25:1047-10-54 Fig. 10: Rat MSC colonies (CFU-F) obtained in vitro in the absence (control) or in the presence of the ODN IMT504

In vivo treatment with IMT ODNs augments the number of Bone Marrow progenitors of MSCs

The bone marrow content of MSC progenitors can be augmented by subcutaneously injecting an animal with IMT ODNs (Fig. 11).

Number of CFU-F / 10x10 6 cells

40 35 30 25 20 15 10 5 0

Control

IMT504

Stem cells (2007);25:1047-10-54 Fig. 11: Bone marrow rat MSC culture after in vivo treatment with IMT504 ODN. Control animals were treated with saline.

In vivo Treatment with IMT504 Augments Recovery of MSCs from Rat Peripheral Blood MSCs progenitors in circulation are a likely natural source of cells for rapid repair of damaged tissues. Table 1 shows that the number of animals with detectable MSCs progenitors is greatly augmented in animals treated with IMT504.

Control Total number animals Animals with detected MSCs progenitors in circulation 12 2

IMT504 12 8

Table 1: Rise in circulating MSCs progenitors after in vivo treatment with IMT504

Proof of Concept (POC) models for tissue repair and protection by treatment with IMT ODNs

1) Bone repair

Stimulation of MSC progenitor expansion by treatment with IMT ODNs in vivo indicates that these ODNs may be used as medicines in the field of Regenerative Medicine, a term currently used to define a number of medical procedures that make use of stem cells to regenerate tissues within damaged organs. To prove this concept, we first chose a rat model of defective bone repair. Fig. 12 illustrates the procedure followed to introduce the experimental defect in the tibial bone of the rat.

B
A: incision performed in the tibia forefront zone, B: Vehicle (3 l) with or without ODN introduced into the defect.

C: radiographic evaluation

Fig. 12: Rat experimental model.

This defect was filled with a gel containing the prototype of IMT ODNs, IMT504, in the group of treated animals and a gel without ODN in the group of control animals. Fig. 13 shows an example of radiological analysis during evolution of the defect in one animal of each group and Fig. 14 the histological analysis of the tibial bone at the site of the defect of one animal of each group after four weeks of treatment.

Control

IMT 504

Week 1

Week 3

Week 5

Data on File Fig. 13: Radiographs after treatment

Control

IMT 504

Data on File

Fig. 14: Histological cuts of rat tibias after a 4-week treatment.

2) Osteoporosis Osteoporosis: Osteoporosis is a progressive systemic disease characterized by a low bone mass and micro-architectural deterioration of bone tissue with a subsequent increase in bone fragility and susceptibility to fracture. Due to its important prevalence worldwide, osteoporosis is considered a serious public health concern. While osteoporosis occurs in men and pre-menopausal women, the problem is overwhelmingly prevalent in postmenopausal women. Currently it is estimated that over 200 million people worldwide suffer from this disease. Approximately 30% of all postmenopausal women have osteoporosis in the United States and in Europe and at least 40% of these women and 15-30% of men will sustain one or more fragility fractures in their remaining lifetime. Ageing of populations worldwide will be responsible for a major increase of the incidence of osteoporosis in postmenopausal women. Osteoporotic fractures are those that occur under a slight amount of stress that would not normally lead to fractures in nonosteoporotic people. Typical fractures occur in the vertebral colum, hip and wrist. The underlying mechanism leading to osteoporosis is an imbalance between bone resorption and bone formation. Either bone resorption is excessive, and/or bone formation is diminished. Bone is manufactured by osteoblast cells, whereas bone resorption is accomplished by osteoclast cells. Trabecular bone is the sponge-like bone in the center of long bones and vertebrae and cortical bone is the hard outer shell of bones. Trabecular bone is more subject to bone turnover, to remodeling and appears more affected in osteoporosis. The main histological alteration in the structure of the osteoporotic bone is a generalized thinning of the trabeculae. Common osteoporotic fracture sites, the wrist, the hip and the spine, have a high trabecular bone to cortical bone ratio and these areas rely on trabecular bone for strength. Ovariectomy induces bone loss in the rat and shares many similarities with postmenopausal bone loss in humans. These include increased rate of bone turnover with resorption exceeding formation and great loss of trabecular bone. Treatment of ovariectomized rats with IMT504 considerably improves bone quality (Fig. 15). According to the diameter and density, trabecular bone appears healthier in the treated animals.

Topics in Tissue Engineering, Vol. 3. Fig. 15: Histological cuts of rat tibias after a 4-week treatment.

3) Multiple Organ Failure in Sepsis

Sepsis is a major cause of mortality and morbidity throughout the world. Despite the use of potent antimicrobial agents and advanced supportive care, the mortality rate of patients with Gram-negative bacteraemia remains high (between 20 and 35%). The annual incidence of severe sepsis is approximately 3.0 cases per 1,000 of the population, over 18 million cases worldwide each year.

The sepsis patient is typically critically ill and requires immediate attention to avoid rapid deterioration. The cost for survivors is high with long-term quality of life being significantly lower than that of the general population.

Severe sepsis is commonly accompanied by multiple organ dysfunction or failure. In order to investigate the organ protection potential of IMT504 rats were infected with Escherichia coli (O45K1H10) and treated with intramuscular injections of IMT504. Fig. 16 illustrates the presence of tissue damage in the liver of a control (untreated) rat and the absence of damage in the liver of an IMT504 treated animal.

Control Data on File

IMT504 treated

Fig. 16: Liver protection during sepsis by IMT504 treatment

4) Neural damage Neuropathic pain refers to a heterogeneous group of pain conditions characterized by lesion or dysfunction of the normal sensory pathways. It is associated with a variety of etiologies, including trauma, infection, diabetes, immune deficiencies, ischemic disorders, and toxic neuropathies. Approximately 26 million patients are affected worldwide and the lifestyle of these patients can be severely affected, a problem compounded by the lack of efficacy and frequent incidence of side effects associated with current treatment options. Opioids are frequently ineffective in treating neuropathic pain. Current treatments, such as gabapentin (NeurontinTM), involve non-selective regulation of neurotransmitter systems or ion channels and generally result in significant dose-limiting CNS side effects. In this area where no current single drug treatment is effective in more than 50% of patients, novel therapeutic approaches are an urgent priority. The hallmarks of neuropathic pain are chronic allodynia and hyperalgesia. Allodynia is defined as pain resulting from a stimulus that ordinarily does not elicit a painful response (eg. light touch) and hyperalgesia is defined as an increased sensitivity to a normally painful stimuli. Progressive tactile hypersensitivity (PTH) manifesting after sciatic nerve crush is a rodent experimental model of neuropathic pain. Rats subjected to an experimental

sciatic nerve crush and treated with intramuscularly injected IMT504, in contrast with untreated rats, do not develop allodynia (Fig.17) as measured by the Von Frey test.

30

Von Frey hair threshold (g)

Control IMT504
20

10

0 0 5 10 15 20 25

Time (days after lesion)

Data on File Fig. 17: Neural protection by IMT504 treatment

5) Diabetes mellitus Adults with diabetes have heart disease death rates approximately 2-4 times higher than adults without diabetes. Other complications also lead to increased morbidity and mortality rates: Risk for stroke, pregnancy-related deaths, end-stage renal disease and blindness. Approximately 60% -70% of persons with diabetes have neuropathy, and more than 50% of the lower limb amputations occur among persons with diabetes.

Daily medication regimens, insulin injection, and blood glucose monitoring are complex and uncomfortable. Reducing morbidity and mortality and improving quality of life for persons with diabetes is a critical public health objective. Streptozotocin is an antibiotic with diabetogenic properties which is widely utilized as a method for inducing diabetes mellitus in experimental animals.

10

Rats subjected to this experimental diabetes show normal levels of glucose in blood after treatment with intramuscularly injected IMT504, in contrast with untreated rats (Fig. 18).

Plasma Glucose Concentration (mg/dL)

800
NORMAL RATS DIABETIC RATS + IMT 504 DIABETIC RATS

600

400

200

0 0 5 10 15 20 25 30

Time (days)
Data on File Fig. 18: Pancreatic function normalization by IMT504 treatment

11

IMT ODNs in preclinical trials

Toxicity of the IMT leader product ODN IMT504 was assessed in preclinical trials performed in several animal species. These trials included: 1) Acute toxicity studies in mice, rats and monkeys 2) Sub-acute toxicity studies in rats and monkeys 3) Local tolerance studies in rats, rabbits and dogs 4) Mutagenicity studies in bacteria and human lymphocytes 5) Pyrogenicity studies in rabbits Results of these studies are described in an attached paper.

Intellectual property

IMT ODNs and their uses are protected by the following patents and patent applications 1. Immunostimulatory oligonucleotides and uses thereof (Immunotech S. A.). i. US Patent 7,038,029 issued on Mar 2nd 2006 ii. US Patent 7,381,807 issued on June 3th 2008 iii. PCT- (WO 03/101375 A2; May 30th, 2003, iv. WIPO; Priority date with patent CA-2388049; May 30th, 2002, Canada); granted. v. ARGENTINA: Sep 27th, 2005; P050104073 vi. AUSTRALIA: Nov 25th, 2004; AU: 2003250334; vii. BRASIL: Nov 29th, 2004; Br.018070017817; viii. CANADA: Nov 12th, 2003; CA: 2487904; ix. EUROPE: Dec 27th, 2004; EP: 03755959.8; x. INDIA: Nov 29th, 2004; In: 3773/DLNP72004; xi. ISRAEL: Nov 29th, 2004; Il: 165466; xii. JAPAN: Nov 29th, 2004; JP: 2004-508733; xiii. KOREA: Nov 30th, 2004; KO:10-2004-7019524; xiv. MEXICO: Nov 30th, 2004; PA: /a/2004/011937; xv. NEW ZEALAND: Nov 25th, 2004; NZ: 536962; xvi. PHILIPPINES: Nov 25th, 2004; PH: 1-2004-502009

12

2. Osteogenic oligonucleotides and WO2006034956; Applied 27/09/2004.

uses

thereof

(Immunotech

S.

A.).

i. ARGENTINA: Feb 6th, 2006; P060100338; ii. AUSTRALIA: Mar 26th, 2007; AU: 2005-289033; iii. BRASIL: Nov 29th , 2004; Br.018070017817; iv. CANADA: Mar 26th, 2007; CA: 2581665; v. EUROPE: Mar 20th, 2007; EP1807061; vi. ISRAEL: Mar 26th, 2007; Il: 182198; vii. JAPAN: Mar 23th, 2007; JP: 2007-532883; viii. KOREA: Apr 4th, 2007; KO:10-2007-7009476; ix. MEXICO: Mar 27th, 2007; Mx/a/2007/003612; x. NEW ZEALAND: Mar 26th, 2007; NZ: 554031; xi. PHILIPPINES: Mar 26th, 2007; PH: 1-2007-500668; xii. USA: Mar 26th, 2007; 2007/11/663833; 3. Oligonucleotides stimulatory of the mesenchymal stem cell proliferation and uses thereof (Immunotech S. A.). WO2006128885; Applied 2005. Int. Search Report: 2007. i. ARGENTINA: Nov 29th, 2007;P; ii. AUSTRALIA: Nov 30th, 2007; AU: 2006-254162; iii. BRASIL: Dec 3th, 2007; P10613211-1 iv. CANADA: Nov 29th, 2007; CA: 2610413; v. EUROPE: Jun 3th, 2005; EP05104854.4; vi. ISRAEL: Dec 3th, 2007; Il: 187804; vii. JAPAN: Nov 27th, 2007; JP: 2008-514102; viii. KOREA: Feb 1st, 2008; KO:10-2008-7000076; ix. MEXICO: Nov 30th, 2007; Mx/2007/015127; x. NEW ZEALAND: Nov 30th, 2007; NZ: 563931; xi. PHILIPPINES: Dec 3th, 2007; PH: 1-2007-502736; xii. USA: Nov 30th, 2007; 2007/11/921384; 4. Methodos to increase the diversityin vivo and in vitro of antibodies produced against an antigen (Immunotech S. A.). USA . Provisional application. May 14th, 2008. PA/2008/ 61053204.

13

5. Immunostimulatory oligonucleotides and uses thereof (Immunotech S. A.). USA . Divisional Patent application from PCT/EP03/05691. Nov 28th, 2008. DP/2008/ 11/921.384.

Publications
- Elias, F., Flo, J., Lopez, R.A., Zorzopulos, J., Montaner, A., Rodrguez, J.M. 2003. Strong Cytosine- Guanosine- Independent Immunostimulation in Humans and Other Primates by Synthetic Oligonucleotides with PyNTTTTGT Motifs. J.

Immunol.171:3697.

- Elias F, Flo J, Rodriguez JM, Nichilo AD, Lopez RA, Zorzopulos J, Nagle C, Lahoz M, Montaner A. 2005. PyNTTTTGT prototype oligonucleotide IMT504 is a potent adjuvant for the recombinant Hepatitis B vaccine that enhances the Th1 response. Vaccine, 23:3597-603.

- Rodriguez JM, Elias F, Montaner A, Flo J, Lopez RA, Zorzopulos J, Franco RJ, Lenial SP, Lopez Salon M, Pirpignani ML, Solimano J, Garay G, Riveros D, Fernandez J, Cacchione R, Dupont J. Oligonucleotide IMT504 induces an immunogenic phenotype and apoptosis in chronic lymphocytic leukemia cells. Medicina (B. Aires). 2006; 66:916.

- Lopez RA, Zorzopulos J. Vaccine shortage for pandemic influenza: can it be solved? Vaccine. 2006; 24:2701.

- Montaner AD, De Nichilo A, Elias F, Rodriguez JM, Flo JM, Lopez RA, Zorzopulos J, Frank R. Ganglioside GM1-binding peptides as adjuvants of antigens inoculated by the intranasal route. Vaccine. 2006; 24:1889-96.

- Rodriguez JM, Elias F, Flo J, Lopez RA, Zorzopulos J, Montaner AD. Immunostimulatory PyNTTTTGT Oligodeoxynucleotides: Structural Properties and Refinement of the Active Motif. Oligonucleotides. 2006; 16: 275-85.

14

- Insua, A.H., Montaner, A.D., Rodrguez, J.M., Elias,F., Flo, J., Lopez, R.A., Zorzopulos, J., Hofer, E.L., Chasseing, N.A. 2006. IMT504, the Prototype of the Immunostimulatory Oligonucleotides of the PyNTTTTGT Class is a Potent Signal for Mesenchymal Stem Cells Expansion In Vitro and In Vivo with Potential Use in Medicine in Tissue Repair Therapy. Stem cells (2007);25:1047-10-54.

- Insua, A.H., Montaner, A.D., Rodrguez, J.M., Elias,F., Flo, J., Lopez, R.A., Zorzopulos. 2007. PyNTTTTGT Oligonucleotides as Tools in Tissue Repair

Procedures. Topics in Tissue Engineering, Vol. 3. Eds. N. Ashanmakhi, R. Reis & E. Chiellini.

Presentations to Scientific Meetings

-Montaner A., Rodriguez J.M., Elias F., Fl J., Lpez R.A. and Zorzopulos J. A novel group of very potent (non-CpG) immunostimulatory oligonucleotides. Sixth Annual Conference on Vaccine Research. National Foundation for Infectious Disease. May 2003. Arlington, Virginia. USA.

-Montaner A., Elias F., Juan Fl, Lpez R.A., Zorzopulos J. Rodriguez J.M. Strong CpG Independent Immunostimulation in Humans and other Primates by Synthetic Oligodeoxynucleotides with PyNTTTTGT Motifs. Sixth Annual Conference on Vaccine Research. National Foundation for Infectious Disease. May 2003. Arlington, Virginia. USA.

-Rodriguez J.M., Elias F., Fl J., Lpez R.A., Montaner A. and Zorzopulos J. Potent (non-CpG) Immunostimulatory Oligonucleotides. 90th Anniversary Meeting,

Federation of American Societies for Experimental Biology. The American Association of Immunologists. May 2003. Denver, Colorado. USA.

-Elias F., Fl J., Lpez R.A., Zorzopulos J., Montaner A. and Rodriguez J. Immunostimulation in Primates by Synthetic Oligodeoxynucleotides with PyNTTTTGT Motifs. 90th Anniversary Meeting, Federation of American Societies for Experimental

15

Biology. The American Association of Immunologists. May 2003. Denver, Colorado. USA.

-Elas F., Montaner A., Rodrguez J, Denichilo A., Fl J, Lpez R. and Zorzopulos J. PyNTTTTGT Oligonucleotide IMT504 is a Potent Vaccine Adjuvant. Experimental Biology. XXX International Congress of Physiological Sciences. March 2005. San Diego, CA. USA

-Rodrguez J.M., Elas F., Lpez Saln M., Montaner A., Franco R.J., Genial S.P., Pirpignani M., Fl J., Lpez R.A. and Zorzopulos J. Immunostimulatory PyNTTTTGT Oligonucleotide IMT504 Induce an Immunogenic Phenotype and Apoptosis in Chronic Lymphocytic Leukemia Cells: Synergy with IL2 and Independency of the Mutational Status of the Ig VH Genes. Experimental Biology. XXX International Congress of Physiological Sciences. March 2005. San Diego, CA. USA

-Rodrguez J.M., Marchisio J., Lpez Saln M., Elas F., Montaner A., Fl J., Lpez R.A. and Zorzopulos J. Induction of high levels of GM-CSF Secretion in Human Peripheral Blood Mononuclear Cells by non-CpG Immunostimulatory

Oligonucleotides. Experimental Biology. XXX International Congress of Physiological Sciences March 31. April 2005. San Diego, CA. USA -Montaner A.D., Elias F., Rodrguez J.M., Flo J., Lopez R. and Zorzopulos J. PyNTTTTGT Oligonucleotide IMT504 is a Potent Vaccine Adjuvant. Eighth Annual Conference on Vaccine Research. May 2005. Baltimore, Maryland. USA.

- Montaner A.D., Hernando Insa A, Rodriguez J, Elas F, Fl J, Hofer E, Lpez R, Chasseing N, Zorzopulos J. IMT504 the Prototype of the Immunostimulatory Oligonucleotides of the PYNTTTGT Class Is a Potent Signal for Mesenchymal Stem Cells Expansion. 4th ISSCR Annual Meeting International Society for Stem Cell Research. June 29-July 1, 2006, Toronto- Canada

16

Conclusions

IMT ODNs are potent stimulators of MSC precursor cells expansion in vitro and in vivo. Therefore, they can be used for regenerative tissue treatments. Since IMT ODNs act on the patients own stem cells, they are completely free of immunological rejection and free of ethical issues related with the use of embryonic stem cells.

POC trials in all these fields are very encouraging.

IMT ODNs are very easy to synthesize and manufacture.

Due to their high solubility and stability in water, they are very promising pharmacologically.

According to performed preclinical studies IMT ODNs are safe drugs.

17