Tel Hashomer Medical Research Infrastructure and Services Ltd. The Technology Transfer Company of Sheba Medical Center
Novel Therapeutics for Human Diseases
Contact : Sylvie Luria PhD. CEO Technology Transfer Company Tel Hashomer Medical Research, Infrastructure and Services Ltd. Tel: +972-3-5305998 Fax: +972-3-5305944 Cell: 052-6667277 [email protected] http://research.sheba.co.il/e/ תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
Sheba Medical Center - Technology Transfer Company Tel Hashomer Medical Research, Infrastructure and Services Ltd. http://rnd.sheba.co.il/
Business: Tel Hashomer Medical Research, Infrastructure and Services Ltd. (THM) the technology transfer arm of Sheba Medical Center, is responsible for managing the intellectual property assets of Sheba Medical Center and to promote the transfer of technologies, innovation and professional know-how for society's use and benefit, and for the development of the medical and health care delivery fields. Sheba Medical Center facilities, experience, human resources and regulations enable the development of a novel idea from its basic science to its product development and prototype, thus rapidly generating value to its IP for commercialization.
Main Activities: Scientific insights and academic breakthroughs often translate into inventions for the benefit of the marketplace. THM bridges the gap between Academia Research and Industry Needs, since the industry is product-based, business-oriented, and focused on time-framed missions, THM helps turn scientific progress into tangible products, while returning income to the inventor and to Sheba Medical Center to support further research. THM receives invention disclosures from faculty, staff and students. We evaluate the innovations for patent applications and develop licensing strategy, consider the technical and market risks. Patentable inventions constitute the majority of THM's licensing activities; however, we also handle collaborations with industrial partners and Tangible Research Property (TRP) such as Tissue Bank, Genomics and Bio-Markers, Cell Therapy, Computational Imaging and more. THM builds a well-structured and organized “value creation” model, as well as several business models pending on industry: (Health IT, Medical Devices, Bio-Medical, ) and on entity (start- up, SME and Big Entity/ Pharma). THM has several strategic support plans such as the “Micro Fund" and strategic collaboration with other research institutes and industry to facilitate invention development.
Royalties Commercialization and Streaming licensing management IP strategy and managemnt
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
THM Strategic Principles to the Success of our Tech Transfer
► We bridge basic research to commercial Value ► We develop close interaction with researchers and industry ► We Build Strategic Capabilities ► We are a “Learning Organization” and Flexible Organization ► We understand the stakeholders need and Value creation ► We Build Collaboration & Alliances ► Our stream: Identify Need from the bedside, Basic and applicable Research- ► We develop broad and Multi-national view
THM Intellectual property's portfolio spans over therapeutics, diagnostics, medical devices and medical tools in the fields of Onco-Genetics, Hemato-Oncology, Epidemiology of Malignant Diseases and Trauma, Lipids, Diabetes, Hypertension, Onco-Surgery , including research in Breast and Colon Cancer, Regenerative medicine, Immunology, Neuro-Immunology, Alzheimer's Disease, Multiple Sclerosis and Psychiatry.
Contact:
Sylvie Luria Ph.D. CEO Technology Transfer Company תל השומר מחקרים תשתיות ושירותים רפואיים בע"מ Tel Hashomer Medical Research, Infrastructure and Services Ltd. Tel: +972-3-5305998 Fax: +972-3-5305944 Cell: +972-052-6667277 [email protected] http://rnd.sheba.co.il/e/ תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
Innovations from Sheba Medical Center - Technologies for Licensing:
Novel Therapy
1. FZD7 - A NOVEL ANTIBODY TARGETING TUMOR STEM CELLS
2. SOMATOSTATIN BASED RECEPTOR TARGETING FOR DRUG DELIVERY IN CANCER TREATMENT.
3. PHARMACEUTICAL COMPOSITION AND METHOD (SIRNA TO STIL GENE) FOR REGULATING
ABNORMAL CELLULAR PROLIFERATION
4. NOVEL PAR-1-BASED THERAPEUTIC COMPOUNDS FOR NEUROINFLAMATORY AND
MALIGNANT DISEASES: DIABETIC NEUROPATHY AND GLIOBLASTOMA MULTIFORME.
5. THE HORMONE KLOTHO: A NOVEL THERAPY FOR BREAST AND PANCREATIC CANCERS
6. NOVEL COMPOSITION, TARGETED-TREATMENT APPROACH FOR MULTIPLE SCLEROSIS
7. NOVEL COMPOSITION FOR THE TREATMENT OF MULTIPLE MYELOMA
8. METHOD AND COMPOSITION OF REPROGRAMMING RENAL CELLS – CELL THERAPY TO
TREAT ACUTE RENAL FAILURE.
9. MULTI-EPITOPE CITRULLINATED PEPTIDE FOR TREATMENT OF RHEUMATOID ARTHRITIS.
10. NOVEL FERTILITY PRESERVATION COMPOSITION TO PREVENT CHEMOTHERAPY & TRANSPLANTATION DAMAGE. 11. A NOVEL NON-HORMONAL COMPOSITION AS CONTRACEPTIVE FOR OVULATION
12. NOVEL COMBINATION FOR HEMOSTATIC-ANESTHETIC DRUG FOR USE IN AMBULATORY
SURGICAL PROCEDURES
13. NOVEL TOPICAL SMALL RNA MOLECULE AS TREATMENT FOR PSORIASIS
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 FZD7 - a novel biomarker for targeting tumor stem cells
Benjamin Dekel , Michal Daniel and Naomi Pode-Shakked, Sheba Medical Center, Israel Categories Antibody, Cancer Therapeutics Development Stage In-Vivo animal model efficacy Patent Status Pending
Background Wnt signaling pathway has been shown to play an important role during human development, taking part in self renewal and differentiation processes of stem cells. Aberrant activation of this pathway is linked to many types of cancer, taking potential parts in disease pathogenesis. For instance, in familial colon cancer, inactivation of the APC protein, a key component of Wnt signaling, is a crucial step in the neoplastic evolution.
FZD7 is a member of the 'frizzled' gene family, encoding 7-transmembrane domain receptors for Wnt signaling proteins. Binding of Wnt proteins to the FZD7 receptor results in activation of the canonical Wnt pathway which usually leads to cell proliferation and survival. FZD7 is known to be over expressed in several malignancies including Colon cancer, Esophageal cancer, Gastric cancer, Wilm's tumor and more, compared to the normal tissue from which each tumor develops. It is becoming clear that many, if not most, malignancies arise from a population of cells that exclusively maintain the ability to self-renew and sustain the tumor via the expression of tumor- progenitor genes. These ‘‘cancer stem cells’’ are often biologically distinct from the differentiated cancer cells that comprise most of the tumor bulk. Since cancer stem cells are believed to be primarily responsible for tumor initiation as well as resistance to chemo- and radiotherapy, their persistence after such treatments may account for relapsing disease and metastasis in many malignancies.
Taking into account the Wnt pathway established role in enabling stem cell properties ('stemness') as well as FZD7 over expression in multiple cancers in which the Wnt pathway is deranged, FZD7 is a potential therapeutic target for numerous malignancies both as a biomarker for cancer stem cells and as a target for attenuating Wnt signaling in these cells which in turn is likely to lead to cancer cell death.
Current Findings Wilms tumor (WT), a pediatric kidney cancer can serve as an excellent model for studying malignant renal stem cells leading to tumor initiation and progression. An important link between aberrant Wnt signaling and WT formation has been established. We have observed frizzled7 (FZD7) up regulation in WT compared to normal kidney and showed FZD7 protein expression on a subset of WT cells. Unexpectedly, incubation of WT cells with anti-FZD7 Ab resulted in apoptosis and cancer cell death, suggesting possible manipulation of the tumor with antibody therapy. Further analysis of additional WTs has identified tumours from which a viable FZD7(+) cell population can be isolated, which enabled characterization of these cells as potential Wilm’s tumor stem cells. Those novel observations highlighted the Wnt pathway and especially FZD7 as a therapeutic target in Wilm’s tumor and possibly in additional FZD7/Wnt dependent תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 malignancies. Initially several anti FZD7 monoclonal antibodies that had potential to induce cell death activity were tested. We have now identified a novel anti FZD7 c-terminus antibody. This Anti FZD7 antibody termed Ab 288-1 was found to inhibit WNT signaling, reduces cell proliferation, and induces cell death in-vitro it malignant cell line cells as well as in primary malignant tissue derived cells.
Objectives 1. Further establishment of anti FZD7 antibody killing effect in Wilm’s tumor in animal models. 2. Assessment of anti FZD7 antibody killing effect in the following additional malignancies in- vivo: i. Melanoma ii. Colon cancer iii. Neuroblastoma
3. Preparation of anti FZD7 humanized antibody with the highest efficiency against tumor stem cells and the best discriminatory capabilities between normal and malignant tissues: 3.i. In vitro and in-vivo assessments efficacy studies with the selected mouse antibody 4. Generation of humanized antibodies 5. in vitro and in-vivo assessments efficacy studies with the selected human antibodiesy
Background of the Invention
It is becoming clear that many, if not most, malignancies arise from a population of cells that exclusively maintain the ability to self-renew and sustain the tumor via the expression of tumor- progenitor genes (Singh et al., 2003; van der Kooy and Weiss, 2000). These ‘‘cancer stem cells’’ are often biologically distinct from the differentiated cancer cells that comprise most of the tumor bulk. Since cancer stem cells are believed to be primarily responsible for tumor initiation as well as resistance to chemo- and radiotherapy, their persistence after such treatments may account for relapsing disease. Recent molecular data obtained by microarrays demonstrate that WTs and WT-stem like xenografts systematically overexpress nephric-progenitor genes corresponding to the earliest stages of normal metanephric kidney development, connecting tumorigenesis and organogenesis in the kidney. In addition to genes that specify the renal lineage, Wnt pathway-related molecules including b-catenin (CTNNB1), frizzled7 (FZD7) and frizzled2 (FZD2), are concomitantly induced. In general, the binding of Wnt ligand to frizzled cell surface receptors normally leads to inhibition of a “destruction complex” consisting of APC/Axin/GSK-3β/Ck1/Dvl and other factors, with subsequent accumulation of dephosphorylated stabilized β-catenin, and transcription of its target genes. Interestingly, recent data have demonstrated an essential role for the Wnt–b- catenin signaling pathway in nephrogenesis and also show a striking link between b-catenin signaling and the development of Wilms tumor. In the latter case, deregulated Wnt signaling with aberrant accumulation of β-catenin in the cytoplasm/nucleus plays an important role. While the Wnt pathway in general is implicated in the maintenance of tissue homeostasis by regulating self-renewal of normal stem cells as well as proliferation or differentiation of progenitor (transit- תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 amplifying) cells, the Wnt receptor FZD7 has been shown to specifically contribute to self- renewal signaling of human embryonic stem cells and to play a significant role in the neoplastic prossess of many types of cancers the most characterize of which is Colon cancer.
In order to establish FZD7 as a marker for stem-like highly malignant cells in WT we attempted to prospectively isolate putative FZD7(+) progenitor cells, in which case incubation of WT cells with an anti-FZD7 Ab resulted in cell death. Additional analysis has now revealed tumors resistant to anti-FZD7 Ab, showing a different intra-cellular route of the Ab-FZD7 complex compared to sensitive tumors and intact cell phenotype. Accumulation of β-catenin was observed exclusively in FZD7-resistant-WT. This coincided with a low sFRP1 and DKK1 (Wnt inhibitors) expression pattern, restored epigenetically with de-methylating agents, and lack of β-catenin mutations. The addition of exogenous DKK1 and sFRP1 to the tumor's cells enabled the sensitization of FZD7-resistant-WT to the FZD7 Ab. Finally, although extremely difficult to achieve due to dynamic cellular localization of FZD7, sorting of FZD7(+) cells from resistant WT, showed them to be highly clonogenic/proliferative, overexpressing WT 'stemness' genes, emphasizing the importance of targeting this cell fraction.
FZD7 antibody therapy alone or in combination with Wnt pathway antagonists may have a significant role in treatment of WT as well as of additional Wnt dependent malignancies.
The Need Despite the major advances in research and development of therapeutic solutions for pediatric solid tumors, there remains an everlasting need for novel regimens which will both reduce the risk of recurrence and resistance of highly malignant disease and decrease the incidence of severe late adverse effects of currently available medications. This problem is easily demonstrated in the case of Wilms tumor (WT). Wilms’ tumor accounts for 6% of all pediatric tumors and is the second most frequent intra-abdominal solid organ tumor found in this patient population. Initial survival rates in the early part of the last century were only 30%, but now long-term survival in both North American and European trials are approaching 85%. While most Wilms' tumors are of low stage, favorable histology, and have a high likelihood of cure with current multimodal therapy, there remains a group of patients whose tumors recur or metastasize for whom intensive salvage regimens result in survival of only 50%. Additionally, WT survivors often suffer from late adverse complications at their second or third decade, resulting from receiving high doses of chemotherapies at a young age. Novel targeted treatment strategies are needed to improve clinical outcomes for children with WT that have unsatisfactory long-term survival, patients of good survival but high potential for late adverse effects due to conventional therapy and a final challenge are those children with both a poor survival and a high potential for late adverse effects. The proposed invention of anti FZD7 antibody for targeting the most malignant cells in the tumor, the cancer stem cells, is a novel treatment approach for pediatric solid tumors as well as for additional malignancies in which Wnt pathway and specifically and FZD7 receptor play a role in disease pathogenesis in addition to marking the tumor stem cells.
Potential Applications As in the case of WT, patients suffering from a variety of solid pediatric malignancies as well as additional Wnt dependent cancers can benefit from the aforementioned therapeutic strategy. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 This suggested novel targeted therapy may prove a significant addition to the armamentarium of current commercially available anti neoplastic medications.
Advantages As opposed to many other antibody dependent targeted therapies in which the target is solely a marker for malignant cells lacking either functional activity against the cell survival mechanism or the specificity to mark the tumor stem cells, targeting the Wnt pathway and specifically FZD7 receptor which play an important role in cell survival as well as being a potential marker for the most malignant cell subpopulation in the tumor, the Cancer stem cells is advantageous due to the fulfillment of both these crucial traits.
Patent COMPOSITIONS AND METHODS FOR TREATING CANCER
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
SOMATOSTATIN BASED RECEPTOR TARGETING FOR DRUG DELIVERY IN CANCER TREATMENT.
Prof. Orenstein, Sheba Medical center, Tel Hashomer
Categories Novel Compositions for cancer drug delivery Development Stage Early preclinical studies Patent Status Pending
Background and Technology Cancer has a tremendous impact on society and it is the second leading cause of death in the U.S. Current cancer chemotherapy therapies are limited with respect to tumor cell targeting, therapeutic windows and the propensity to induce resistant tumor clones. One promising approach to overcome these problems is to use Targeted Drug Delivery (TDD) whereby a carrier that has a specific affinity to cancer cells is linked to an anticancer drug. While this strategy is being extensively investigated, the literature describes very limited range of conjugation methods mainly using mono-functional linkers for the coupling of drugs to carriers as enzymes, antibodies, peptides, and biodegradable polymers. The linkage of several and different drugs to a target specific carrier might improve the therapeutic efficacy of TDDs but this has yet to be achieved. Moreover, the possibility of engineering deviations in time release of different drug moieties from such a multi-conjugate suggests that employing a platform with multiple attachment and controlled release capabilities may represent a significantly improved architecture for drug multilinking – multi-released properties in TDD.
We target the somatostatin receptors: Somatostatin receptors (SSTr) is over expressed and found in neuro-endocrine tumors (NETs) such as lungs, brain, skin, adrenals, pancreas, liver and gastrointestinal tract. Somatostatin (SST) is a small cyclic peptides, secreted by the delta pancreatic cells and at the hypothalamic nerve endings. SST is involved in the regulation of diverse biological processes acting primarily as a negative regulator of a variety of different cell types, blocking processes such as insulin, glucagon and growth hormone secretion, cell growth and smooth muscle contraction. Each of the SST functions is initiated by the binding of the peptide to one or more of the five different receptor subtypes (SSTR1-5) which are expressed on the cell surface and belong to the G coupled protein receptor family. In normal tissues SSTRs present in endocrine and exocrine tissues, neuronal and immune cells, and have been identified in multiple sites throughout the nervous system, gastrointestinal tract, breast and pancreas. In various pathological conditions such as malignancies, autoimmune diseases, inflammation and others, SSTRs are significantly over-expressed in affected organs.
SST analogs: Native SST has a short half-life in vivo (2-3 minutes) due to enzymatic activity. Somatostatin analogs are the first class of receptor binding peptides having gained clinical applications. Long-lasting synthetic somatostatin analogs have been developed and introduced into clinical practice. In addition, Nuclear Medicine based on SSTR tumor targeting were תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 developed as well as radio-labeled SST analogs for diagnostic applications in oncology . Systemic administration of SST analogs labeled with γ-emitting or/and ß-emitting radio-nuclides was introduced for both cancer imaging and therapy. Currently, SSTR-targeted scintigraphy (PET, SPECT) and radionuclide therapy are well established techniques in the routine clinical practice mainly for NETs. Recent studies showed that a vast variety of human malignancies including lung, breast, brain and prostate cancer tend to over-express one or several SSTRs compared to adjacent normal tissues. This gives prospect for broader clinical applications of SST analogs during the next decade.
The Need for Novel SST Analogs. Despite the progress in the development of SSTR analogs, there is still a need for improvement of the receptor targeting peptides by increasing the specific binding capacity to wider SSTR subtypes, prolonging binding and intracellular internalization processes and decreasing adverse effects on healthy tissues. These include the optimization of peptide analog properties e.g. receptor specificity and pharmacokinetics in order to obtain better selectivity of tumor and drug accumulation within tumor cells in comparison to normal tissues, especially to the critical organs such as liver, kidney and spleen. For example, the classical radioactive somatostatin analog DOTA-Tyr3-octreotide (DOTA-TOC) has relatively rapid clearance from the tumor and high uptake by kidney that makes it less safe and effective for radiotherapy.
Our Innovations:
We have designed, tested and patented a novel family of SST analogs (PTR peptides) for tumor targeting applications using a proprietary backbone cyclization technology. A large number of derivatives were studied in vitro and in vivo, and one of them, (PTR 3207-86) labeled with a fluorescent moiety (FITC), showed very high tumor-targeting properties with no toxic effects. This leading compound displayed distinctive pharmacokinetic profile with clearance from the normal tissues while being accumulated in malignancies selectively. In mice xenografts models bearing human lung and colon carcinomas significant differences in fluorescence intensity between tumor and normal tissues were observed during the period from 24 to 72 hours after intravenous administration of the agent. Laser scanning microscopy revealed membrane binding with internalization of the agent into cancerous cells as well as in the peri- and intra-tumoral blood vessels.
We are developing the therapeutic platform for novel targeted chemotherapeutic conjugates for advanced cancer treatment based on somatostatin receptor specific drug delivery.
We have developed novel cancer targeting carriers – backbone cyclized somatostatin (SST) analogs (PTRs). A number of novel analogs were found to show outstanding SST receptor specific binding in vitro and tumor accumulation in vivo with distinctively high and long-lasting tumor-to-normal tissue ratio (TNTR) compared to currently clinically used analogs. These preliminary results provide an excellent opportunity to use PTRs as specific carriers in TDD concept for advanced selective cancer therapy. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
Our carrier is conjugated via unique chemical structure component to chemotherapy drug to enhance treatment selectivity, safety and clinical output .
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 COMPOSITION AND METHOD FOR REGULATING ABNORMAL CELLULAR PROLIFERATION
Prof. Izraeli Sheba Medical Center, Tel Hashomer
Categories Novel siRNA composition to treat solid tumors Development Stage In-vivo efficacy studies Patent Status pending
Background and Technology The STIL (SIL) gene, SCL (Stem Cell Leukemia TAL1 Interrupting Locus), located on chromosome 1, was cloned from the most common chromosomal rearrangement in T cell Acute Lymphoblastic Leukemia. It encodes a 150 kDa cytosolic and centrosomal protein conserved in vertebrates. STIL is tightly regulated during the cell cycle. It is expressed only in proliferating cells, reaching a peak in mitosis during which the protein is phosphorylated and then degraded upon entrance to the next cell cycle. The critical requirement of STIL for cell growth, proliferation and survival during embryonic development, and its regulation during the cell cycle suggest that STIL might have a role in tumorogenesis. We have demonstrated increased expression of STIL RNA and protein in a wide variety of cancers. Further analysis revealed that genes with expression patterns similar to STIL were mitotic checkpoint genes regulated by the transcription factor E2F1 known to be activated in proliferating cells. The mitotic index is increased with expression of STIL and its expression in epithelial cancers is associated with metastases and poor prognosis. Inhibition of STIL expression by a specific anti STIL siRNA in colon cancer cells was established and demonstrated - STIL blocks mitotic entry and causes apoptosis of this colon cancer cells in-vitro and in-vivo. Similar results have been obtain on a variety of cancer cells representing the most common types of human cancer (Cervical, lung, breast, prostate, gliomas ,renal cancer, ovarian cancer and leukemias) using anti STIL siRNA oligonucleotides. The lethal effect of the knockdown of the human SIL is completely rescued by transfection of the murine STIL, demonstrating the specificity of the siRNA mediated approach. Most importantly we have recently shown that intravenous administration of STIL siRNA packed in neutral liposomes is effective alone and in combination with cisplatinum in an orthotropic xenograft model of human ovarian cancer. STIL is not necessary, however, for survival of all normal proliferating cells as we have mouse embryonic stem cells and fibroblasts that are negative for the STIL gene. Importantly, the unique sensitivity of cancer cells to anti-mitotic drugs gives these treatments an excellent therapeutic ratio with relatively few side effects. We conclude that STIL is a novel target for treating abnormal proliferation and cancer effective in vitro and in vivo models for variety of human cancer models. . Anti STIL activity was demonstrated to be effective in inhibiting tumor cell development
We have also demonstrated its efficacy in Malignant primary brain tumors, in particular high grade gliomas, are uniformly fatal within months from diagnosis despite multi-modal aggressive therapeutic approaches. Chemotherapy carries only a limited benefit due to poor penetration of תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 most therapeutic agents across the blood-brain barrier. Accordingly, local delivery of therapeutic agents may provide a significant therapeutic advantage in brain tumor patients. Convection- enhanced drug delivery (CED) is by far the most promising potential delivery platform that is available for efficient local distribution of drugs within tumors. Successful implementation of CED is expected to have a significant impact on patient outcome as it would deliver high concentration of the therapeutic agent with little or no systemic toxicity over large volumes of tissue. However, the use of CED is currently limited by the challenge to produce optimal drug formulations, the design of appropriate means to monitor the convective process, methods to enhance drug distribution while minimizing toxicity and most importantly by the lack of effective anti Glioma targeted therapies. As local therapy is a highly relevant approach for treatment of gliomas and as STIL is expressed in gliomas but not in normal brain, we hereby propose a preclinical study that integrates a highly effective delivery system, CED, with novel siRNA based targeted therapy. Our results suggest that a specific siRNA to STIL inhibited tumor growth by approximately 50%. In rat glioma where treatment is applied on established tumors these are significant results. As in-vitro data clearly demonstrates synergism between STIL siRNA DNA damaging agents such as Cisplatinum (that are used for glioma therapy with modest effects) we recommend testing such combination therapy in future experiments. Potential Applications SiRNA is a promising emerging research field directed at the development of future therapies, currently in initial clinical studies. The application of siRNAs for specifically silencing various disease-associated alleles has been demonstrated in animal models. SiRNA can be directed against various types of cancers and the proposed research program is designed to study the application of siRNA in combination with Carboplatin, delivered by CED, for the treatment of glioma. The outcome product would be a novel formulation for the treatment of gliomas consisting of high dose siRNA against STILL, low dose Carboplatin, a solvent for increased drug distribution and Gd-DTPA for real-time treatment monitoring. A successful proof of concept using this formulation may lead to additional formulations with various combinations of siRNAs and other therapeutic agents for the treatment of hematopoietic and solid tumors including CNS malignancies Advantages The critical requirement of STIL for cell growth, proliferation and survival during embryonic development, and its regulation during the cell cycle suggest that STIL might have a role in tumorogenesis. The fact that STIL is not indispensable for normal proliferating cells survival makes it a good target for treating abnormal proliferation and cancer. We have validated STIL gene as a target for cancer therapy. Development Stage STIL siRNA anti-proliferative effect was demonstrated in cancer cell lines and primary tumors models in animals for colorectal cancer, ovarian cancer and glioblastoma. Patent 1 siRNA sequences – Pending 2 CHFR DERIVED PEPTIDES TO STIL AS ANTI-CANCER COMPOUNDS
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 The Market The four main types of cancer therapy are chemotherapy, hormone therapy, target therapy and immunotherapy. Of these, the target therapy segment has the largest share of the market. Worth an estimated $22.9 billion in 2008, this segment will reach $69.1 billion in 2013, for a CAGR of 24.7%. Target therapy comprised approximately 45.0% of the overall market in 2007. This figure is expected to increase to 62.5% in 2013. An estimated 22,280 new cases of ovarian cancer were diagnosed in the US in 2012. The Ovarian Cancer Drug Market is set to More Than Double to $1.6 Billion by 2022. Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor with survival measured in months from diagnosis. In 2009, 22,070 new cases of primary brain tumors were diagnosed in the USA and 12,920 patients died of primary brain tumors. An estimated 103,170 cases of colon cancer and 40,290 cases of rectal cancer were newly diagnosed in the UAS in 2012. Colorectal cancer is the third most common cancer in both men and women.
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
Novel PAR-1-based Therapeutic Compounds for Neuroinflammatory Diseases
Prof. Chapman, Sheba medical Center, Tel Hashomer
Categories Novel composition to treat human diseases Development Stage Animal models in-vivo efficacy Patent Status Provisional-pending
Background and Technology Many neurological diseases are known to be associated with increased inflammation and over activation of protease activated receptors (PAR) by protein factors of the coagulation cascade. We suggest a coagulation-based protease inhibitor potentially affecting mostly neuronal PAR as a novel intervention site for neurological disease. Thrombin is a protease well known for its role in coagulation. In addition to its role in the coagulation cascade, thrombin affects many cellular events by a group of receptors named protease activated receptors (PARs) PAR-1 to PAR. PAR-1 has been found in many tissues including the brain where it is found on several cell types including neurons, astrocytes and microglia. PAR-1 is localized to the non-compacted myelin of Schwann cells at the node of Ranvier and in the neuromuscular junction it is localized to the peri-synaptic glia and the post- synaptic muscle. We were the first to show a functional role for PAR-1 activation on the glia component of the node of Ranvier in the PNS where activation of the receptor caused conduction block. PAR-1 activation was shown to hold a physiological role at the CNS synapse where its activation modulates synaptic transmission by causing LTP and seizure-like activity and potentiates the synaptic NMDA receptor. PAR-1 is a marker of glial structures adjacent to the most physiologically active parts of the nervous system, the synapse and the node of Ranvier. Activation of these structures through PAR-1 has profound effects on electophysiology and therefore these glial structures and especially the PAR-1 receptors on them are a novel and highly promising target for therapy in disease. We have collected many significant findings supporting the role of PAR-1 activation in mediating neurological dysfunction in disease. We have found that thrombin-like activity was elevated in sciatic nerves derived from animal models of both Guillain-Barré Syndrome (GBS) (EAN) and diabetic neuropathy (STZ). In both models a significant decreased PAR-1 level was found together with increased physiological thrombin-inhibitors (PN-1, PN-2) indicating that the natural response of the nervous system in these diseases is to increase thrombin inhibition. A relatively general thrombin inhibitor, N alpha-tosyl-L-lysine chloromethyl ketone (TLCK), as well as the PAR-1 antagonist (SCH79797) were found to increase nerve conduction and improve neuropathy associated clinical deficits of these GBS and diabetes model animals significantly. The Need Neuropathy is a general term indicating a disease which affects peripheral nerves. The most common cause of neuropathy is diabetes mellitus, occurring in 60% of all diabetic patients. Diabetic peripheral neuropathy (DPN), one of the most prevalent forms of diabetes-related neuropathy, principally manifests itself as sensory loss and weakness of the lower limbs and ultimately accounts for significant morbidity contributing to amputation. In the PNS the תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 prototypical inflammatory disease affecting nerve cells is the Guillain-Barré syndrome (GBS). GBS is a group of acute inflammatory neuropathies and its animal model is experimental autoimmune neuritis (EAN). Acute inflammatory demyelinating polyneuropathy (AIDP) is the most common form of GBS, and is characterized by demyelination of peripheral axons and axonal degeneration in severe cases. The mechanism leading to GBS are still being elucidated and most of the evidence points to an immune attack at the node of Ranvier.
The Market GlobalDatas analysis suggests that the global Diabetic Neuropathy(DN) market was worth $ 1.41 billion in 2010. It is forecast to grow at a Compound Annual Growth Rate (CAGR) of 8.5% for the next seven years, to reach $ 2.49 billion by 2017. This is primarily attributed to the increase in the prevalence of the disease and the strong pipeline with new first-in-class therapies. The global diabetic neuropathy therapeutics market is attractive, with high unmet need. This unmet need in the market is around 41%, which is approximately valued at $ 0.58 billion. The unmet need is due to the lack of availability of effective treatment options targeting disease progression, difficulties in diagnosis and the moderate safety profile of the marketed drugs. The efficacy of the marketed products is moderate. The current treatment options focus on symptomatic relief and are only moderately efficacious. The DN market offers opportunity for newer disease modifying treatment options. GlobalData analyzed the current competitive landscape for DN drugs and found that the competition is moderate to weak. There are only two FDA (Food and Drug Administration) approved products which are Cymbalta (duloxetine) from Eli Lilly and Lyrica (pregabalin) from Pfizer but these are targeting diabetic neuropathy pain only. In addition, gabapentin and amitriptyline are also used as off label drugs for the treatment of pain associated with diabetic neuropathy. Generics such as topical capsaicin are also used but such products do not hold a major share in the market. The other available drug classes for the treatment of the pain associated with DN are antidepressants, antiepileptics, opioids and NSAIDs (Non-Steroidal Anti- Inflammatory Drug). The competition is weak due to a lack of competing products for Cymbalta and Lyrica. The market has a huge potential for molecules with better safety and efficacy profile and especially such molecules targeting disease progression and offering a cure for DN. Potential Additional Applications Neurodegenerative diseases (NDD) are destined to become the next great health crusade over the next several decades. This will support sustainable long-term sales growth of NDD products: the collective total sales of the eight markets in 2013 will be over $10bn by 2013. In the US alone, neurodegenerative disease accounts for more than $1.3 trillion of nondiscretionary health-care spending. Advantages Our strategic approach has been to assume that PAR-1 over-activation plays a central role in many neurological diseases and to specifically target this pathway in order to find novel, more efficient and less toxic therapies since the compositions are targeted to a physiological driving disease target. Development Stage We have designed new compositions, based on the thrombin binding sequence of PAR-1, targeting the increased protease activity found in disease states and specifically protects תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 PAR-1. The approach is based on the fact that receptor activation needs the specific binding of thrombin and similar proteases to the receptor. This composition target specifically PAR-1 which seems significantly different from the equivalent sequences in PAR-2, 3 and 4. Preliminary studies are highly encouraging in the potency of the compounds to treat neuropathy diseases such as diabetic neuropathy GBS. We hypothesize that these compounds can be further improved by altering the length of the PAR-1 mimicking peptide sequence and also by certain modifications such as in the prolyl group which we have found to be susceptible to in the normal brain. Patent “PAR-1-based Therapeutic Conjugates and Uses Thereof" application, 2013, pending.
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 Novel PAR-1-based Therapeutic Compounds for ALS
Prof. Chapman, Sheba medical Center, Tel Hashomer
Categories Novel composition to treat human diseases Development Stage Animal models in-vivo efficacy Patent Status Provisional-pending
Background and Technology Many neurological diseases are known to be associated with increased inflammation and over activation of protease activated receptors (PAR) by protein factors of the coagulation cascade. We suggest a coagulation-based protease inhibitor potentially affecting mostly neuronal PAR as a novel intervention site for neurological disease. Thrombin is a protease well known for its role in coagulation. In addition to its role in the coagulation cascade, thrombin affects many cellular events by a group of receptors named protease activated receptors (PARs) PAR-1 to PAR. PAR-1 has been found in many tissues including the brain where it is found on several cell types including neurons, astrocytes and microglia. PAR-1 is localized to the non-compacted myelin of Schwann cells at the node of Ranvier and in the neuromuscular junction it is localized to the peri-synaptic glia and the post- synaptic muscle. We were the first to show a functional role for PAR-1 activation on the glia component of the node of Ranvier in the PNS where activation of the receptor caused conduction block. PAR-1 activation was shown to hold a physiological role at the CNS synapse where its activation modulates synaptic transmission by causing LTP and seizure-like activity and potentiates the synaptic NMDA receptor. PAR-1 is a marker of glial structures adjacent to the most physiologically active parts of the nervous system, the synapse and the node of Ranvier. Activation of these structures through PAR-1 has profound effects on electophysiology and therefore these glial structures and especially the PAR-1 receptors on them are a novel and highly promising target for therapy in disease. We believe that ALS involves the selective dysfunction and loss of astrocytes associated with large pyramidal neurons. These astrocytes are involved in the metabolism of glutamate, have NMDA receptors and are also clearly defined by the presence of PAR-1 on their processes. We have collected many significant findings supporting the role of PAR-1 activation in mediating neurological dysfunction in this disease. We have found an increased thrombin-like activity in brains of an ALS animal model (SOD-1) and decreased PAR-1 levels. A relatively general thrombin inhibitor, N alpha-tosyl-L-lysine chloromethyl ketone (TLCK), as well as the PAR-1 antagonist (SCH79797) were found to increase the life span of these ALS model animals significantly. Thus excessive levels of thrombin activity and activation of PAR-1 may play a significant role in the pathogenesis of ALS and thus represent a target for intervention. The Need Amyotrophic lateral sclerosis (ALS) is one of the most common motor disease and most rapidly progressive neurodegenerative diseases of unknown cause. Until today there is only one drug that slows disease progression. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 To our understanding PAR-1 can be used at least as a diagnostic marker for the astrocyte function and disease progression of ALS patient and it also may serve as a therapeutic target in ALS treatment.
The Market The Market for Amyotrophic Lateral Sclerosis (ALS) Amyotrophic lateral sclerosis (ALS) is a neurological disease and also termed as Lou Gehrig’s disease. The disease affects the motor neurons, which control the action of voluntary muscles, as a result the messages between brain and muscle fibers are blocked. ALS could occur in a person due to genetic inheritance, mutation in gene which encodes enzyme copper-zinc superoxide dismutase coupled with defect in chromosome 9. This disease is more prevalent in males. All these abnormalities results in the release of glutamate which damage the motor neurons controlling voluntary muscles. ALS projects various symptoms such as spasticity, cramps, fasciculation, respiratory problems and muscles weakness. ALS results in the inability to breathe; this is due to non- functioning of diaphragram and chest muscles caused due to muscle weakness It has been estimated that every year around 5,000 people in U.S. are diagnosed with amyotrophic lateral sclerosis. The ALS market was worth $150m in 2010 across the seven major markets. With the only approved treatment for ALS, Sanofi-Aventis’s Rilutek (riluzole), facing generic incursion in 2013, The unmet need for ALS treatments with a novel mechanism of action, which can alter disease progression, remains high. There are few late-stage therapies in the pipeline: potential launches to 2019 include, Avicena’s ALS-02 (2015) and Mitsubishi Tanabe’s Radicut (edaravone) (Japan only, 2013). Advantages Our strategic approach has been to assume that PAR-1 over-activation plays a central role in many neurological diseases and to specifically target this pathway in order to find novel, more efficient and less toxic therapies since the compositions are targeted to a physiological driving disease target. Development Stage We have designed new compositions, based on the thrombin binding sequence of PAR-1, targeting the increased protease activity found in disease states and specifically protects PAR-1. The approach is based on the fact that receptor activation needs the specific binding of thrombin and similar proteases to the receptor. This composition target specifically PAR-1 which is significantly different from the equivalent sequences in PAR-2, 3 and 4. Preliminary studies are highly encouraging in the potency of the compounds to inhibit brain thrombin and thus potentially treat ALS. We hypothesize that these compounds can be further improved by altering the length of the PAR-1 mimicking peptide sequence and also by certain modifications such as in the prolyl group which we have found to be susceptible to in the normal brain. Patent “PAR-1-based Therapeutic Conjugates and Uses Thereof" application, 2013, pending תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
THE HORMONE KLOTHO: A NOVEL THERAPY FOR BREAST AND PANCREATIC CANCERS
Ido Wolf, Tamar Rubinek, Bella Kaufman, Lilach Abramovitch, H. Phillip Koeffler, Chaim Sheba Medical Center, Israel
Background of the Invention Pancreatic cancer is a fatal disease and the 5 years survival of patients who suffer from advanced disease is less than 10%. Current treatment for metastatic pancreas cancer consists of chemotherapy and is unsatisfactory, with a median survival following chemotherapy of about 6 months. Breast cancer, in which malignant cells develop in the tissues of the breast, is the most common cancer among women, and the second most common cause of cancer death in women in the United States. Klotho, a transmembrane protein, is expressed in the brain, kidneys and pancreas. It is also expressed in various sex-hormone responsive organs including placenta, testis, prostate and ovary. Klotho can be shed and act as a hormone, and has been identified previously as an inhibitor of the insulin growth factor (IGF)-1 pathway in hepatocytes and muscle cells.
Our data suggest high levels of klotho protein expression in normal breast samples and normal pancreatic cells, but very low expression in cancerous breast cells or cancerous pancreatic cells. In addition, noted inhibition of breast cancer cell growth following over-expression of klotho protein, and growth enhancement of klotho protein-expressing cells following klotho protein knock-down; and revealed modulation of the IGF-1 and the insulin pathways by klotho protein. Taken together, the results suggest klotho protein as a novel breast cancer tumor suppressor. Moreovere, we have found additive effects of klotho and various chemotherapy agents and noted more than 80% inhibition of PANC-1 cells by the combination of klotho and low dose of chemotherapy. Animal studies demonstrate a therapeutic effect of Klotho. Treatment of mice with Klotho protein resulted in a significant dose response inhibition of tumor growth. The present invention relates to the use of the klotho protein for the treatment and diagnosis of cancer, such as breast cancer and pancreatic cancer as well as other IGF-1 dependent cancers. Recently, we have characterized the specific and selective domain of the protein, with selective anti-tumor, including a unique mutation introduced in the active domain, which enhanced its activity and specificity.
The Need Pancreatic cancer is responsible for 6% of cancer death each year. Pancreatic cancer has the highest fatality rates of all cancers, and is the 4th highest cancer killer in the US among both men and women. Each year about 33,000 individuals in the US are diagnosed with this condition and more than 60,000 in Europe. The only therapy currently available for metastatic pancreatic cancer is standard chemotherapy which adds about 1 to 2 months to the median survival time. Breast cancer is the leading cause of cancer-related mortality among women worldwide. In 2007, more than 470,000 cases of the disease were diagnosed in the seven major pharmaceutical markets. High unmet needs still persist for this tumor type. Despite recent drug approvals in the adjuvant and metastatic setting, the overall survival remains below five years. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 The breast cancer (BC) market is a steadily growing market due to effective treatments which have decreased mortality and resulted in a longer duration of therapy. The five-year survival rates for women with BC have increased from under 70% 50 years ago to around 90% now. In 2007, we estimate that sales grew by 22% compared to 2006, generating revenue of over US$11.3 billion. Branded therapies were responsible for 92% of this total (US$10.5 billion). Breast cancer is the most common cancer among women.
Cancer can only be positively diagnosed by biopsy, which involves removal of a small tissue sample. A substantial need remains for a more accurate and less invasive means of determining the presence of cancer. Klotho blood tests can use for diagnosis.
Potential Applications Klotho is a potentially potent and specific therapeutic agent for the treatment of cancer patient. Administration of Klotho in combination with standard chemotherapy may greatly approve cancer patient survival. Monitoring blood levels of Klotho for early diagnosis of cancer and prognosis. The major potential applications are breast and pancreatic cancer
Advantages Protein / peptide drugs pose unique advantages over traditional small molecule drugs as they are targeted and specific for cancer types. Given the importance of detecting cancer early in its development, and the potential harm that may occur as a result of false positives or false negatives from mammography and other techniques for screening of various types of cancer, a substantial need remains for a more accurate and less invasive means of determining the presence of cancer. Moreover, currently used techniques cannot predict cancer development among patients with benign breast cancer. Development Stage In vitro as well as in vivo studies have proved the concept of Klotho and its active domain as a potent diagnostic and therapeutic agent for cancer. Structure function analysis of Klotho protein resulted in the active extra cellular subunit with the anti cancer activity. Biochemical studies of Klotho and its domain, in terms of structure function analysis to dissect its cellular mechanism of action and its anti-tumor activity. Patent 1. KLOTHO PROTEIN AND RELATED COMPOUNDS FOR THE TREATMENT AND DIAGNOSIS OF CANCER 2. NEW COMPOSITION OF KLOTHO PROTEIN AND RELATED COMPOUNDS FOR THE TREATMENT AND DIAGNOSIS OF CANCER
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 NOVEL COMPOSITION, TARGETED-TREATMENT APPROACH FOR MULTIPLE SCLEROSIS
Prof. Anat Achiron and Dr. Michael Gurevich, the Center for Multiple Sclerosis, Categories Novel Drug for Multiple Sclerosis (MS) Development Stage Preclinical Patent Status Three related patent applications(2011, 2013, 2014) Background Multiple sclerosis (MS) is the most common demyelinating (Damage to the myelin sheath around nerves) disease of the central nervous system (CNS) affecting young adults. The peak disease onset is between 20 to 40 years of age, more prevalent in females and is the third leading cause for disability after trauma and rheumatic diseases in young adults. MS has a characteristic geographical distribution, more likely to occur in northern Europe, the northern United States, southern Australia, and New Zealand than in other areas. Disease prevalence in USA is 120/100,000 (250,000 to 350,000 cases) and in Israel about 60/100,000. The main pathologic findings in MS is the presence of infiltrating mononuclear cells, predominantly T lymphocytes and macrophages, that surpass the blood brain barrier and induce an active inflammation within the brain and spinal cord, attacking the myelin and resulting in gliotic scars and axonal loss. Thus, the multiple inflammatory foci, plaques of demyelination, gliosis and axonal pathology within the brain and spinal cord contribute to the clinical manifestations of neurological disability. The acute and chronic inflammatory processes can be visualized by brain and spinal cord MRI as hyperintense T2 or hypointense T1 lesions. The etiology of MS is not fully understood. The disease develops in genetically predisposed subjects exposed to yet undefined environmental factors and the pathogenesis involved autoimmune mechanisms associated with autoreactive T cells against myelin antigens. It is well established that not one dominant gene determines genetic susceptibility to develop MS, but rather many genes, each with different influence, are involved. The initial pathogenic process that triggers the disease might be caused by one group of genes, while other groups are probably involved in disease activity and progression. MS is subdivided into several clinical subtypes; when it first presents by new onset of neurological symptoms affecting the CNS and accompanied by demyelinating lesions on brain magnetic resonance imaging (MRI), it is defined as probable MS. A diagnosis of relapsing- remitting (RRMS) definite MS is made when a subject defined as probable MS experiences a second neurological attack. The course of RRMS, which occurs in 85 % of patients, is characterized by attacks during which new neurological symptoms and signs appear, or existing neurological symptoms and signs worsen. Usually an attack develops within a period of several days, lasts for 6-8 weeks, and then gradually resolves. During an acute attack, scattered inflammatory and demyelinating CNS lesions produce varying combinations of motor, sensory, coordination, visual, and cognitive impairments, as well as symptoms of fatigue and urinary tract dysfunction. These symptoms are a direct result of the body's own immune system attacking the myelin layer that surrounds the brain and spinal cord. The outcome of an attack is unpredictable in terms of neurological squeal, but it is well established that with each attack, the probability of complete clinical remission decreases, and neurological disability and handicap are liable to develop. In about 15 % of patients the disease has a primary progressive course, characterized by gradual onset of neurological symptoms that progress over time, without any attacks. This course appears mostly in patients with disease onset above the age of 40 years and more often in males. The only course of MS in which treatment was effectively established is RRMS. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 Using high-throughput gene expression microarrays and advanced bioinformatics analytical tools such as supervised learning and feature selection algorithms, we identified gene targets related to the POL-1 pathway that characterize benign MS (BMS) and are correlated with good clinical outcome. BMS is a non-active disease pattern of MS, a preferable clinical variant of RRMS in which patients remains fully functional neurologically without significant disability's assessed by the Expanded Disability Status Scale (EDSS) and get a score lower than 3.0 within 10 years after disease onset. About five to 10 percent of MS patients have a benign course. Unfortunately, it takes as long as 15 years after the initial diagnosis to know if a course is benign. We have studied the molecular events associated with BMS and identified the significant involvement of POL-1 pathway and suggested it's role in BMS and in RRMS patients with good clinical outcome as the regulation inhibitor that inhibition in the regulation of MS disease activity by suppression of inflammation and enhancement of apoptosis of autoreactive lymphocytes.
The Technology Applying high throughput gene expression microarrays we identified that the suppression of polymerase 1 (POL1) pathway is associated with benign course of multiple sclerosis (MS). This finding supports the rationale for direct targeting of the POL1 transcription machinery as an innovative strategy to suppress MS. To evaluate the effects of a specific POL1-I we used experimental autoimmune encephalomyelitis (EAE) mouse model. The effect of POL1-I treatment was modulated through decreased expression of POL1 pathway key-related genes LRPPRC, pre-RNA, POLR1D and RRN3 together with activation of P53 dependent apoptosis of CD4+ splenocytes. Our findings demonstrate that POL1 pathway inhibition delayed and suppressed the development of EAE and ameliorated the disease in mice with persistent clinical signs. We have demonstrated that inhibition of this pathway have therapeutic effect in animal model and in X-vivo blood cells. We have designed a unique molecule that inhibit specifically the POL1 pathway. We have demonstrated its efficacy in EAE animal models The Need There is currently no cure for MS, and the therapies available are effective only partially. The current MS FDA approved disease modifying drugs (DMDs) include a large list of drugs [Avonex (interferon beta-1a; Biogen Idec) Betaseron (interferon beta-1b; Berlex, Inc./Schering), Copaxone (Glatiramer Acetate; Teva), Rebif (interferon beta-1a; MerckSerono), Gilenia (Novartis) and Tysabri (Natalizumab; Biogen Idec)], all have shown efficacy in reducing relapse rate by 29% to 53%; however their effect on the progression to irreversible disability is minor. These DMDs are associated with various adverse events ranging from local injections site reactions to life threatening events associated with JCV infection and PML, and thus have prompted a new wave of R&D heralded by the top biopharmaceutical companies with the production of new innovative MS related therapies such as BG-12 (Biogen Idec), Alemtuzumab (Genzyme), Teriflunomide (Sanofi-Aventis) and Laquinomod (Teva). Still, even with the new therapies the effects are not maximal and moreover, there are no molecular markers that can identify immediate response to therapy. Patients are treated for long periods before the assessment of response to therapy can be made, mainly based on clinical response and imaging data. In addition, most of the DMDs are administered intravenously, intramuscularly or subcutaneously and are associated with discomfort to the patients that lead to reduce adherence תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 to the treatment and therefore sub-optimal clinical response. Accordingly, there is a need for highly effective immunomodulatory treatment that will inhibit disability progression and will lead to eradication of MS and that the monitoring and response to this treatment will be available at face value within a short period of time. The Market The global market for Multiple Sclerosis (MS) disease-modifying products reached $10.1 billion in 2012 and nearly $10.9 billion in 2013. This market is expected to grow to nearly $14.2 billion in 2018. Small molecules, as a segment, reached nearly $4.6 billion in 2013. The segment is projected grow to nearly $8.9 billion in 2018. Today there are currently eight disease-modifying therapies (DMTs) for MS approved throughout the world. Biologics represent the largest class of MS disease-modifying therapies by sales and by number of approved therapies, accounting for six out of the eight approved MS disease-modifying products (Avonex, Betaseron, CinnoVex, Extavia, Rebif and Tysabri). Of these biologics, five are beta interferon products. All of these approved MS agents are able to slow down the progression of the disease but none can reverse any nerve damage that occurs due to demyelination. The initial 2004 Food and Drug Administration (FDA) approval of Biogen Idec's and Elan's Tysabri marked the approval of the first therapeutic for MS that may actually reverse some of the effects of MS. Disease-modifying agents to treat MS are currently one of the largest classes of pharmaceutical products by sales in the U.S. and throughout the world. Sales of these agents for MS have grown from less than $1 billion worldwide in 1999 to around $11 billion in 2013. New approaches to treating MS are being pursued by pharmaceutical and biotech companies as many of them look to move beyond agents such as the interferons that primarily slow the progression of MS. These potential new disease-modifying agents with novel mechanisms of action or improved delivery systems will provide powerful new options for patients failing or intolerable to the currently prescribed treatments for MS. The dominance of first-line injectable DMTs, including the interferon beta (IFNß) agents: Bayer’s Betaseron/Betaferon (IFNß-1b), Biogen’s Avonex (IFNß -1a) and Merck’s Rebif (IFNß- 1a), and Teva’s Copaxone (glatiramer acetate), has been a salient feature of the MS therapeutics market. However, the competitive landscape is undergoing significant change with the emergence of oral therapies, several pipeline products with notable efficacies, and looming generics/biosimilars following the patent expiries of key branded products during the forecast period. In addition, the entry of new companies such as Sanofi/Genzyme and F. Hoffmann-La- Roche/Genentech will challenge the position of the established players in the MS marketplace. IP Status Three patents regarding this technology were filed:
1- RNA Polymerase I Inhibitors and Uses Thereof – Application umber 61/910,060 (28.11.2013). 2- Methods of Predicting Clinical Course and Treating Multiple Sclerosis – Application number 13/260,573 (28.03.2011). 3- RNA Polymerase I Inhibitors and Uses Thereof
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NOVEL COMPOSITION FOR THE TREATMENT OF MULTIPLE MYELOMA
Dr Merav Laibo , Sheba Medical Center Categories Novel Drug , Multiple Sclerosis Development Stage Preclinical Patent Status Three related patent applications(2011, 2013, 2014) Background and Technology Multiple myeloma (MM), also known as plasma cell myeloma, myelomatosis, or Kahler's disease, is a cancer of plasma cells, a type of white blood cell normally responsible for producing antibodies. In multiple myeloma, collections of abnormal plasma cells accumulate in the bone marrow, where they interfere with the production of normal blood cells. Most cases of multiple myeloma also feature the production of a paraprotein—an abnormal antibody which can cause kidney problems. Bone lesions and hypercalcemia (high blood calcium levels) are also often encountered. Multiple myeloma is diagnosed with blood tests (serum protein electrophoresis, serum free kappa/lambda light chain assay), bone marrow examination, urine protein electrophoresis, and X- rays of commonly involved bones. Multiple myeloma is considered to be incurable but treatable. Remissions may be induced with steroids, chemotherapy, proteasome inhibitors, immuno-modulatory drugs such as thalidomide or lenalidomide, and stem cell transplants. Radiation therapy is sometimes used to reduce pain from bone lesions. We have unfolded the cellular mechanism by which plasma cells became tumorogenic , and have identified a small lead molecule for the development of new drug to treat MM. Number of New Cases and Deaths per 100,000: The number of new cases of myeloma was 6.1 per 100,000 men and women per year. The number of deaths was 3.4 per 100,000 men and women per year. These rates are age-adjusted and based on 2007-2011 cases and deaths. Lifetime Risk of Developing Cancer: Approximately 0.7 percent of men and women will be diagnosed with myeloma at some point during their lifetime, based on 2009-2011 data. Prevalence of this cancer: In 2011, there were an estimated 83,367 people living with myeloma in the United States.
Multiple myeloma is the second most prevalent blood cancer (10%) after non-Hodgkin's lymphoma. It represents approximately 1% of all cancers and 2% of all cancer deaths. Although R1
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 the peak age of onset of multiple myeloma is 65 to 70 years of age, recent statistics indicate both increasing incidence and earlier age of onset. DEVELOPMENT STAGE: We have studied our composition with respect to mechanism of action and in animal tumor bearing model with new modality of action: ► The molecule reduces viability and significantly decreased survival of several human MM cells of MM cells regardless of sensitivity to conventional chemotherapy: ► The molecule induces apoptosis and inhibits proliferation of MM cell in a time- and concentration-dependent manner. ► The analysis of the novel compound on cell signalling have been revealed and was shown to affect the AKT pathway. ► Our compound enhances cytotoxicity of conventional and novel anti-MM therapies. ► In vivo anti-MM efficacy of the novel composition was evaluated in a xenograft murine model. Over 80% reduction of the tumor volune was observed' with dose response effect. The Need Till date there is no cure for MM and the survival rate is about 46%. Initial treatment of multiple myeloma depends on the patient’s age and comorbidities. In recent years, high-dose chemotherapy with autologous hematopoietic stem-cell transplantation has become the preferred treatment for patients under the age of 65. Treatment for multiple myeloma is focused on therapies that decrease the clonal plasma cell population and consequently decrease the signs and symptoms of disease. Treatment with bisphosphonates (e.g. pamidronate or zoledronic acid) are routinely administered to prevent fractures; they have also been observed to have direct anti- tumor effect even in patients without known skeletal disease. Advantages Our research results opens the opportunity do developed a novel drug that correspond to the direct target in the plasma tumor cells. It is a kinase inhibitors and orally-bioavailable. We believe that this might lead to less side effects and decrease mortality rate. The Market In 2012- 2013 two important new products reach the market, Onyx’s Kyprolis and Celgene’s Actimid. Both of these have advantages over current drugs, but they’re basically incremental improvements on what we already have – proteasome inhibitors and immuno-modulators. The market can be categorized on the basis of products by major manufacturers such as Thalomid by Celgene Corporation, Doxil by Johnson and Johnson, Revlimid by Celgene Corporation, and Velcade by Johnson & Johnson. The market is segmented on the basis of geography into the North American, Asia-Pacific, European and Rest of the World (RoW) regions The market for multiple myeloma therapeutics is expected to grow with rising incidences of the disease amongst population. The American Cancer Society (ACS) has estimated that by the end of 2014, nearly 24,050 new cases of multiple myeloma will be diagnosed in the U.S. The Multiple Myeloma Drug Market will experience robust 5.6 % annual growth from 2010 to 2020, sales will reach $9.9bn in 2017. Future outlook Currently we are expending the in – vivo efficacy models and evaluate the therapeutic window and safety issues. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
The Multiple Myeloma Drug Market Will Experience Robust 5.6 Percent Annual Growth from 2010 to 2020, sales will reach $9.9bn in 2015.
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METHOD AND COMPOSITION OF REPROGRAMMING RENAL CELLS
Prof. Dekel, Sheba Medical Center, Tel Hashomer
Categories Cell Therapy, Renal Failure Development Stage Preclinical Patent Status 3 Patent applications - Pending
Background and Technology The kidney is an essential organ which removes waste products and blood and regulates intravascular volume, electrolyte and acid-base homeostasis (PH). the degenerative condition which causes loss of kidney function is called chronic kidney disease (CKD), chronic renal disease or chronic renal failure. The most common causes of renal disease are diabetes, high blood pressure and obesity. Other causes include genetic kidney disease, congenital kidney malformations, inflammatory kidney disease or manifest as idiopathic kidney disease (unknown cause). The disease can also appear following: a traumatic injury with significant blood loss, the abrupt decrease of blood flow to the kidneys, damage to the kidneys following sepsis (a severe blood infection), hindered urine flow (oftentimes caused by an enlarged prostate), harm caused by drugs or toxins and pregnancy- related complications CKD has been classified into five main stages; stage one is the mildest, stage five is the harshest. Stages one and two may go unnoticed while stages three, four and five tend to cause irreversible damage to the kidneys. No specific therapeutics are available to CKD When CKD reaches a terminal phase it is termed end stage renal disease (ESRD). This disease stage requires renal replacement. The current treatments for kidney failure are: Dialysis: hemodialysis – a procedure usually done at dialysis centers three times a week in which an external filter is used to process your blood and the clean blood is returned to your body. peritoneal dialysis – a daily procedure that can be done at home in which the lining of your abdominal cavity, that holds organs like the stomach, intestines and liver, is used to filter your blood. Transplantation: kidney transplants are a solution for end-stage renal disease but remain a challenge due to the shortage of organs.
We are utilize the principles of kidney regeneration for discovery of novel and feasible therapeutic options. We recreate kidney tissue; using stem and progenitor cells with the aim of repairing patients' kidneys by prevention of disease progression to its terminal phases. The Technology and Product תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 We have developed two main processes which aim to grant Chronic Kidney Disease patients (CKD) alternative, permanent solutions to curing this ailment. We will offer patients the option of avoiding the very cumbersome, time-consuming dialysis treatments given at hospitals and the stressful and often inaccessible option of renal transplantation; leaving many patients without a viable kidney replacement until end-stage renal disease has taken their lives. Our two processes use nephron-forming progenitor cells in order to repair diseased ones. They are both in pre-clinical stages and when clinically tested and approved, have potential to restore function in those suffering from kidney disease in a long-term, non-surgical manner. The first process uses healthy allogeneic, or foreign nephron-forming progenitor cells which are expanded in culture and bio-banked. These healthy cells, also known as adult kidney stem cells which have the intrinsic, pre-programmed capacity to form mature kidney tissue and rejuvenate the diseased tissue are to be trialed by injection into the bodies of renal failure sufferers. The ready-to-use stem cells can be shipped to patients just as a medication can be transferred. As these are foreign cells immunosuppression may be required to protect them from being rejected. However, in some kidney diseases patients are already taking immunosuppressive medication and paving the way for allogeneic cells. The second process uses autologous, or internal patients' own nephron-forming progenitor kidney cells. Your body has the ability to generate new kidney tissue which is then extracted and expanded upon in order to allow for renal tissue regeneration. These cells can be obtained by a renal biopsy. The new, healthy cells generated are then placed into kidney disease patients’ bodies; allowing for healthy kidney-related activity to resume. Whether kidney patients opt for the allogeneic or the autologous route to curing renal failure, our processes have potential for ground-breaking answer to the reformation of healthy kidneys and to a long, disease-free life.
Potential Applications
• CELL THERAPY: The cells could be targets for therapies to combat not just acute kidney disease but also chronic and degenerative diseases, including congenital disorders of the kidney where tissue generation or regeneration is needed. • DRUG DISCOVERY: Cell lines carrying specific kidney diseases would accelerate research into renal pathology as well as provide a system to evaluate molecules or protocols for their therapeutic potential. • TISSUE ENGINEERING: Kidney stem cells, grown and differentiated in vitro, could be incorporated in next-generation dialysis devices.
Advantages Cell transplantation has been identified as a potential alternative to renal transplantation. It has been proven that nephron-forming cells grown ex vivo (outside of the body) are able to reconstitute into functional renal structures in vitro and in vivo. We have developed methodologies needed to allow for the formation of renal structures, kidney regeneration and improved functional outcome. We use novel human cell types, specific growth conditions, innovative tissue and genetic engineering techniques, and original imaging strategies. Combining the outcomes of these technologies has accelerated development of renal cell therapy for renal תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 failure. Use of the proposed cell source for tissue regeneration provides a reliable and sufficient supply of cells that can be used for cell-based therapies specific to renal failure. Development Stage Our technology was implemented in an In-Vivo experimental animal models for efficacy and safety. Patent Three patents in methods and composition - Pending
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 MULTI-EPITOPE CITRULLINATED PEPTIDE FOR TREATMENT OF RHEUMATOID ARTHRITIS
Howard Amital, Smadar Gertel and Yehuda Shoenfeld, Sheba Medical Center, Tel Hashomer
Immune tolerance induction with multi-epitope citrullinated Categories peptide attenuates arthritis manifestations Development Stage Pre-clinical Patent Status Patent pending
Background and Technology Citrullinated peptides are major targets of disease-specific autoantibodies in Rheumatoid Arthritis (RA). The formation of citrullinated peptides elicits an immune response that lead to tolerance breakdown manifested by production of anti-citrullinated protein antibodies (ACPA). Currently, citrullinated peptides serve as a biomarker for the diagnosis of RA by measuring ACPA titers in patient's sera. The accumulations of citrullinated proteins at inflammation sites suggest them as targets for tolerance induction. Since a diversity of citrullinated autoantigens exist in RA we tailored a multi-epitope citrullinated peptide (Cit-ME), derived from sequence of the most prevalent citrullinated autoantigens. The ability of the Cit-ME peptide to induce tolerance following administration to arthritic rats was studied. Treatment of Adjuvant Induced Arthritis with Cit-ME reduced significantly the disease severity compared to untreated rats. Moreover, amelioration of disease manifestations was shown by increased regulatory T cells and apoptosis rate and reduced Th17 cells. We also found that RA-patient's derived PBMC incubated in the presence of Cit-ME peptide elicited up-regulation of the TGF-β gene expression that was associated with an increase in CD4+FoxP3+ T regulatory cell and elevated T cells apoptosis rate. In addition it reduced the expression of the pathogenic cytokines INF-γ and TNF-α and the proportion of Th-17 cells. Thus, the use of citrullinated peptides-based immunotherapy may be a promising approach for tolerance induction in experimental arthritis and perhaps even in susceptible individuals that are ACPA seropositive in human arthritis. Moreover, immunotherapy based on citrullinated peptides could be given as preventive treatment to susceptible ACPA+ population as personalized medicine prior to disease onset. In an attempt to develop a preventive treatment given to delay or prevent arthritis symptoms onset in susceptible ACPA sero-positive individuals prior to arthritis course. The Need Currently, citrullinated peptides serve as a biomarker to identify ACPA presence and titers in patient's sera. Our innovative approach is to employ the citrullinated peptides as immune tolerance inducing agents that will assist to reverse the abnormal immune response in RA. To the best of our knowledge this study is the first to employ citrullinated peptides as tolerogenic agents for immunomodulation and restoration of the autoimmune response in RA. Potential Applications The aim of this study is to elucidate essential mechanisms of immune tolerance induction with citrullinated peptides. Successful achievement of our tasks will provide a new horizon for future development of immune therapy based on citrullinated peptides to suppress the autoimmune response in arthritis. The advantages of tolerance induction-based approach over other general immune modulating therapies, relies in its effective suppression of the specific-autoreactive תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 immune responses, without widely affecting the entire immune system as most available treatments do. The citrullinated peptides used in this study derived from endogenous protein sequences expressed exclusively in diseased targeted tissues in arthritic state and not in healthy tissues. In addition the advantage of administration of a multi-epitope peptide over whole protein is less susceptibility to produce autoantibodies against a peptide due to their relatively low molecular weight. Advantages Our innovative approach is to employ the citrullinated peptides as tolerance inducing agents that will assist to restore the abnormal immune response in RA. To the best of our knowledge this study is the first to employ citrullinated peptides as tolerogenic agents for immunomodulation and restoration of the autoimmune response in RA. Future development of immunotherapy based on immune tolerance based on citrullinated could be given to prevent or delay arthritis onset to susceptible subpopulation diagnosed as ACPA sero- positive individuals prior to arthritis symptoms that could develop arthritis in the near future. Development Stage The project is currently ongoing in two phases: 1) Treatment of experimental models of arthritis with Cit-ME peptide. We aim to perform next the following experiments: a) Optimal dose-dependent of the Cit-ME peptide for amelioration of arthritis manifestations. b) Mode of peptide administration (S.C., oral, intra nasal). c) Comparison to conventional therapy (such as methotrexate (MTX)). d) Potential use of Cit-ME as combinational therapy. e) Cit-ME peptide as prophylactic treatment for arthritis.
2) Mechanisms of tolerance induction by the Cit-ME peptide. Analysis of immunomodulation by the Cit-ME peptide in ex-vivo experiments with RA-derived cells (synovial cells, PBMC).
Patent "Synthetic Peptides for the Treatment of Autoimmune Diseases" PCT/IL2014/050243 The invention relates to synthetic peptides, including peptides comprising a plurality of epitopes, each epitope being derived from a different protein, and peptides comprising a plurality of citrullinated residues. The present invention also relates to use of said peptides for the treatment of autoimmune diseases and disorders.
Potential Market The prevalence of RA is estimated to be 0.8% worldwide, with women twice as likely to develop the disease as men. In the United States, RA afflicts 1.3 million people with a further 126,000 new diagnoses each year. It is responsible for 250,000 hospitalizations and 9 million physician visits each year. According to Global Data, the RA therapeutics market was valued at $10.3 תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 billion globally in 2010, and has doubled over a four-year period after growing at a Compound Annual Growth Rate (CAGR) of 12.3%2. More than 80% of drug sales are biological agents although these are more often used as second line therapy due to their high cost.
Drugs used for treatment of RA are classified as first line agents- consisting of non steroidal anti- inflammatory drugs (NSAIDs) and steroids such as glucocorticoids and second line agents- consisting of disease modifying anti-rheumatic drugs such as chloroquine, gold salts, penicillamine, cyclosporine, and several biological agents like TNF-α and interleukin-1 inhibitors.
Global rheumatoid arthritis market is driven by increase in incidences of RA patients, global ageing population and increasing healthcare expenditure by government of various nations. Factors which are affecting pharmaceutical companies are pricing pressure, patent expiries of RA drugs, high costs of treatments involved and escalating R&D costs.
The pharmaceutical market for arthritis generated over $15 billion in revenues in 2012.
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NOVEL FERTILITY PRESERVATION COMPOSITION TO PREVENT CHEMOTHERAPY & TRANSPLANTATION DAMAGE
Prof. Dror Meirow, Sheba Medical Center, Israel
Categories Pharmacological Composition, Fertility Preservation, Woman Health Development Stage Preclinical Patent Status Pending
BACKGROUND AND TECHNOLOGY One of the most significant long-term sequelae of exposure to cytotoxic drug treatments is infertility, secondary to premature ovarian failure (POF) or insufficiency. As a result, protection of the ovarian reserve and prevention of infertility has become the primary quality of life issue facing patients and their physicians. Current options for female fertility preservation in the face of cytotoxic treatments include embryo, oocyte and ovarian tissue cryopreservation. However, these methods are limited by patient age, status or available timeframe before treatment and necessitate invasive procedures. Improvements in current fertility preservation options as well as development of new, non-invasive options are imperative. Pharmacological agents that can prevent the loss of follicles at the time of treatment are an attractive approach that would provide significant advantages over existing fertility preservation techniques in that they would be suitable for patients of all ages and life stages, would not require invasive surgical procedures or subsequent use of assisted reproductive technologies, and would prevent the myriad endocrine related side effects of POF other than infertility. We have recently demonstrated the molecular mechanism by which chemotherapy causes primordial follicle loss in mice. We have characterized the pathway by which chemotherapy triggers activation of the dormant follicle population as well as inducing death of the growing follicles, thereby depleting the ovarian follicle reserve. Since the growing follicles are responsible for maintaining the dormancy of the follicle reserve by producing the negative regulator, Anti Mullerian Hormone (AMH), the death of the growing follicles results in a reduction of AMH follicle suppression allowing mass activation of the dormant follicles. Activation and loss of the dormant follicle reserve occurs not only with chemotherapy treatment, but also in other situations which disrupt the normal microenvironment of the ovary, such as ovarian tissue cryopreservation and transplantation –one of the current options for female fertility preservation. Our data opens up new avenues for potential therapeutic prevention of follicle loss during chemotherapy and ovarian tissue cryopreservation. AMH and the cellular pathways involved in regulation of follicle activation represent potential targets for the development of new, effective, non-invasive Fertility Preservation options.
DEVELOPMENT STAGE Our studies and data support the following preclinical statements: The ovarian insufficiency and follicle loss that follows chemotherapy and ovarian tissue grafting is, to a significant degree, a result of the activation and growth of the dormant follicle reserve. Follicle dormancy and activation are controlled by both intracellular (PI3K pathway) and extracellular (AMH) mechanisms, both of which provide targets for pharmacological תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 intervention to prevent the loss of follicles caused by chemotherapy and ovarian tissue transplantation. AMH reduces the follicle activation and loss that occurs with chemotherapy and with ovarian tissue transplantation. AMH is a potential effective, non-invasive therapeutic for Fertility Preservation.
FUTURE OUTLOOK
Currently we are completing the AMH optimal dosage and timing response in in-vivo animal models. We are generating an alginate gel containing AMH for coating ovarian tissue grafts, and assessment of AMH diffusion into the graft. We are evaluating the use of gel coating containing AMH to reduce follicle loss in ovarian tissue grafts in combination with chemotherapy and following transplantation.
THE NEED Cancer treatment is associated with significant morbidity, of which gonadal toxicity is a pertinent and common long-term side effect of curative chemo-radiotherapy. Chemotherapy has been shown to cause a dramatic loss of the ovarian primordial follicle reserve often resulting in POF and infertility in female cancer patients. Chemotherapy with alkylating agents, such as cyclophosphamide, is particularly ovotoxic and can deplete the ovarian follicle reserve significantly. While younger women and those who received lower cumulative doses of chemotherapy may regain menstrual periods, these may not occur regularly and may cease prematurely due to the reduction in follicle reserve, resulting in premature menopause.
A DVANTAGES Prevention of follicle loss via therapeutic agents has inherent advantages over currently available methods of fertility preservation, being that it is suitable for patients of all ages and life stages, does not require invasive surgical procedures or subsequent use of assisted reproductive technologies, and additionally would prevent the myriad endocrine related side effects of POF other than infertility. The use of AMH in particular has additional advantages in that it is a naturally occurring endogenous hormone and is therefore unlikely to be toxic or interfere with the anti-cancer effects of chemotherapy.
GLOBAL MARKET ANALYSIS According to the report, “Infertility Drugs and Devices Market – Global Analysis, Size, Share and Forecast, 2010-2017,” the overall infertility drugs and devices market was estimated at $3.55 billion in 2010 and is expected to reach an estimated value of $4.77 billion by 2017, growing at a compound annual growth rate of 4.3 percent from 2012 to 2017. The US fertility market is estimated to be between 3-4 Billion USD, comprising of fertility medications and assisted reproductive technologies and services. The fertility pharmaceutical sector, estimated to be 1.5 billion USD, primarily consists of drugs that induce ovulation. The demand for fertility services and drugs is expected to grow by 4-6% in the next several years, primarily driven by the following key growth drivers: Aging and infertility Increasing prevalence of obesity תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 Significant increase in the prevalence of cancer survivor's rate especially due to the use of aggressive chemotherapies. Chemotherapy is also widely used in non-cancer patients such as autoimmune diseases. Cultural shifts The National Institutes of Health estimated the overall costs of cancer in 2010 at $263.8 billion, of which $102.8 billion was for direct medical costs (total of all health expenditures), $20.9 billion for loss of productivity due to illness and $140.1 billion for cost of lost productivity due to premature death. In 2004, Medicare payments for all Part B drugs for medical oncology reached a total of $5.3 billion (of $2.3 billion for chemotherapy and $1.5 billion for erythroid growth factors). A 2007 study said that drugs prescribed by oncologists account for more than 40% of medicare spending, and spending on oral chemotherapy drugs as a proportion of total pharmacy benefits costs more than doubled between 2002 and 2006, increasing from 0.3% to 0.7%.
Sales of cancer drugs in general doubled between 2005 and 2010. The firm Natixis estimates that the industry will grow at 8% per year, reaching $93 billion in 2016, and IMS health estimated that they will grow at nearly double the rate of the global pharmaceutical market, potentially reaching $80 billion by 2012. IMS predicted a growth rate of 12-15%, due to expensive new treatments, increasing numbers of patents in major markets, as well more people in emerging markets gaining access to modern targeted therapies. The chemotherapy market contains hundreds of competitors, ranging from large brand name marketers to small niche generic producers, making it very competitive. The company with the largest market share is Sanofi-Aventis, at 33.9% in 2008. It offers a variety of anti-cancer products, including the leading drugs Eloxatin, with reported sales of $2 billion in 2008, and Taxotere, with reported sales of $3 billion that year. Eli Lilly & Company also has a strong presence in the chemotherapy market with Gemzar and Alimta. Schering-Plough, with the popular products Temodar and Caelyx, has an 8.8% market share.
It is important to note that chemotherapy drugs are also used to treat autoimmune diseases, and that these patients are also a target for fertility preservation. Currently, 80 to 100 autoimmune diseases have been identified, but about 40 other conditions have a suspected autoimmune basis. Common autoimmune diseases include rheumatoid, psoriatic arthritis, psoriasis, systemic lupus erythematosus, multiple sclerosis (MS), myasthenia gravis, inflammatory bowel disease (Crohn’s disease and ulcerative colitis), sarcoidosis, and scleroderma.
IP S TATUS
" METHODS FOR PREVENTING PREMATURE FOLLICLE ACTIVATION" - PENDING A composition and method, for preventing premature follicle activation and loss induced by medical treatment thereby preserving fertility in a subject.
תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 A Novel Non-Hormonal Composition as Contraceptive for Ovulation
Ariel Hourvitz, Yuval Yung, Svetlana Markman and Gil Yerushalmi, Sheba Medical Center
Categories Women Health , Contraceptive Composition, Fertility, Non Hormonal Development Preclinical studies – efficacy and safety analysis. Stage Patent Status Methods for Controlling Fertility- 2015 , Pending
BACKGROUND AND TECHNOLOGY Prostaglandins (PGs) are a group of hormone-like lipid compounds, are organic anions, that are derived enzymatically from fatty acids and have important physiological functions PGs are local mediators of a variety of biological and pathological processes, where their biological effects include triggering inflammation, fever and pain; induction of labor; modulation of renal hemodynamics and of water and solute reabsorption; and cause constriction or dilation in vascular smooth muscle cells. Further, PGs have long been known to participate in female reproductive functions, including ovulation, fertilization, luteolysis, implantation and parturition. As such, PGs are known as one of the major intra-ovarian mediators of the ovulatory cascade. Prostaglandin transporter (PGT) mediates PGs transport cross biological membranes. PGT is a 644 amino acids membrane protein, function as a PGs transporter with high affinity for PGE2, PGF2α and PGD2.
Endometrial prostaglandin F2α (PGF2α) is the luteolytic hormone. Cellular transport of PGF2α in the uterine endometrium is critical for regulation of the estrous cycle. PGT regulates the PGF2α efflux and influx in endometrial cells that influence luteolytic mechanisms in ruminants. Our research and studies focus on the effect of PG and Ovulation. Our preliminary results indicate that inhibition of PGT by unique composition and bromocresol green (BCG) blocks ovulation.
Our results provide new insight into mechanisms that regulate the action of PGE2 within the pre-ovulatory follicles, and emphasize the major role of PGT in PGE2 signaling and ovulation. Based on our preliminary results, PGT inhibitors can be developed as non hormonal contraceptive agent.
DEVELOPMENT STAGE 1. We have demonstrated that PGT is expressed in human granulosa cells following induction by hCG. The induction of PGT transcript was among the highest differentially expressed transcripts showing 392 fold inductions in expanded cumulus cells as compared to compact cumulus cells. 2. We have demonstrated in animal studies that ovulation is inhibited by PGT inhibitors.
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T H E N E E D 1) Non hormonal contraceptive agent instead of hormonal contraceptive agents. 2) Non hormonal contraceptive agent for woman that cannot receive hormonal contraceptive agents 3) Non hormonal contraceptive agent for woman who began their menstrual cycle. 4) Non hormonal contraceptive agent for IVF instead of hormonal ovulation inhibition
A D V A N T A G E S There are many contraceptive options available for women today. Most of them are hormonal contraceptive, with the drawback associated with hormonal treatments. Moreover, none block ovulation as the primary method of pregnancy prevention. Selective inhibition of ovulation is an important target for the development of new contraceptives. Ovulation inhibition without alteration of ovarian steroid hormone synthesis would be particularly valuable. This approach would minimize or eliminate the side effects experienced by many women who use current hormonal contraceptives but still provide protection against undesired pregnancy. Moreover, some women are prohibited from taking hormonal contraceptives due to increased risk in developing serious medical illnesses such as heart attack, migraines and blood clots. Thus, providing non-hormonal effective and safe contraceptives for women is important for public health
T H E M ARKET Based on the product types, the global contraceptives market is segmented into two major segments, namely, contraceptive drugs and contraceptive devices. The global major players that are Actavis, Inc., Bayers AG, Church & Dwight, Co., Inc., CooperSurgical, Inc., Mayer Laboratories, Inc., Merck & Co., Inc., Reckitt Benckiser plc, Pfizer, Inc., Teva Pharmaceutical Industries Ltd. and The Female Health Company.
The global market for contraceptive pills is estimated to reach $1.03 billion by the end of 2019.
Most of the contraceptive devices are used in order to avoid to side effects and demanding hormonal contraceptive. The contraceptive devices market revenue alone was more than USD 9 billion in 2013.
In 2014 there were 62 million U.S. women in their childbearing years (15–44). Some 62% of all women of reproductive age are currently using a contraceptive method. In the US , 27.5% of all contraceptive users are using hormonal Pills (combined estrogen and progesterone). תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944
A report published on April 2014 by the WCG (Women Care Global) claimed that globally more than 20 million women in the world are lack access to contraceptive, mainly in developing countries which urge for a high quality non-demanding solution (for ex' one shot a month given by the healthcare organizations in the area)
We estimate a great potential market in special treatment population as IVF treatments and different chronic medications care. Those particular populations enjoy limited options of contraception – mostly devices.
FUTURE OUTLOOK Modification of contraception option.
IP S TATUS Provisional Application was filed by Sheba Medical Center on 20/01/2015 under the title "Methods for Controlling Fertility", Pending
The present invention provides methods and compositions for inhibiting ovulation in a female subject, including a subject in need of contraceptives and a subject undergoing fertility treatments. The methods comprise administering to the female subject at least one prostaglandin transporter (PGT) inhibitor.
Main Claim: A method for inhibiting ovulation in a female subject, comprising administering to said female subject a pharmaceutical composition comprising at least one prostaglandin transporter (PGT) inhibitor.
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NOVEL COMBINATION FOR HEMOSTATIC-ANESTHETIC DRUG FOR USE IN AMBULATORY SURGICAL PROCEDURES
Prof. U. Martinowiz and I. Zilinsky, Sheba Medical Center
Categories Ambulatory Surgical Procedures, decrease in hematoma, better healing Development Stage Clinical trial (phase I/II) Patent Status " ANESTHETIC COMPOSITIONS AND METHODS" pending
B ACKGROUND AND TE C H N O L O G Y Transexamic acid ((TXA=hexakapron)) is a synthetic analog of the amino acid lysine that competitively inhibits the activation of plasminogen to plasmin and is used as anti-fibrinolytic agent. TXA in systemic administration has been shown to reduce blood loss in many surgical fields as cardiac, orthopedic, dental, postpartum hemorrhage, scoliosis, tonsillectomy etc without increase in thromboembolic complications. In trauma it has been shown to reduce not only blood loss but also mortality. In addition to the systemic use of TXA it is used for many years locally in teeth extraction and orthopedic procedures in anticoagulant patients and even in patients with severe coagulation disorders such as hemophilia. TXA is considered to be “first line treatment” in almost every patient suffering from massive bleeding. 1-11 Bleeding during the surgery: masks the surgical field and interferes with hemostasis. increases wound hematoma and hence infections. increases electro cautery use and contributes to tissue damage and less optimal scar quality. lengthens the procedure and intensifies patient's discomfort. delay wound healing.
OUR NOVEL APPROACH Novel combined composition of anesthetic compounds together with a hemostatic agent.
DEVELOPMENT STAGE: Clinical trial (phase I/II) after IRB Approval
T H E M AIN NEED: Use of the suggested technology in patients treated with anticoagulants or antiaggregants As the population is getting older the number of patients requiring anticoagulants and antiaggregants (as atrial fibrillation or orthopedic surgery) is expanding. The development of new oral direct anticoagulants is also responsible for the growing number of patients treated with this drug. The limited knowledge and experience with the new drugs is creating great תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 confusion among the physicians leading to cessation of treatment before surgical interventions in patients with high risk of thromboembolic complications, exposing them to severe or even fatal complications
Advantages of the combined Hemostatic -Anesthetic agent: 1. Improve hemostasis and decrease hematoma, leading to better results and better healing. 2. The use of hemostatic-anesthetic combination will reduce the number of patients that will stop their anticoagulation treatment before surgery and reduce the risk to develop thromboembolic complications. T H E M ARKET Ambulatory surgical and emergency center service offers cost-effective and time saving approach to facilitate variety of ophthalmology, urology, gastrointestinal, orthopedic, vascular, pulmonary, pain management, plastic surgery and other surgeries. Ambulatory surgical process is been increasingly accepted by the patient population as these surgeries provides safe, high quality and innovative approach for surgical interventions. US and European promote ambulatory surgeries and technological advancements, that further boost the market growth of ambulatory surgery in near future. Advancement in operative technique with safety protocols and surgical instruments would enable more complex surgeries such as retinal detachment, disc operation and pars plana vitrectomy to be performed under ambulatory basis hence drives the market growth. United States anesthesia drugs market is expected to be more than US$ 3 Billion by 2018. General Intravenous Anesthesia drugs market is dominating the United States anesthesia drugs market. In further classification of General Intravenous Anesthesia Drugs market, Propofol and Benzodiazepines Class (Diazepam and Midazolam) are the top contributors to intravenous anesthetic drugs in the United States anesthesia drugs market. In classification of Local Anesthesia Drugs Market Share Lidocaine holds the maximum market share. All ambulatory surgical procedures including plastic surgery procedures.
FUTURE OUTLOOK The use of Hemostatic-Anesthetic agent will become a routine procedure in all ambulatory surgeries requiring local anesthesia.
IP S TATUS :
" ANESTHETIC COMPOSITIONS AND METHODS" - priority date- August 2014
S TAGES OF DEVELOPMENT : 1. In vitro tests to evaluate the clot formation and stability of each component and the mixture. 2. Safety/Efficacy evaluation of application of gauze impregnated with hemostatic anesthetic solution on surgical wounds (Mohs operation). 3. Safety/Efficacy evaluation of intradermal injection of the hemostatic anesthetic solution during Mohs surgery. 4. Safety/Efficacy evaluation of intradermal injection of the hemostatic anesthetic solution in various ambulatory surgical procedures.
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NOVEL TOPICAL SMALL RNA MOLECULE AS TREATMENT FOR PSORIASIS
Prof. Sidi, Dr. Avni, Sheba Medical Center, Tel Hashomer
Categories Psoriasis, Topical Treatment , Novel Composiation
Development Stage Obtain Cell culture results, establishing mouse model, preliminary results in mouse model. Patent Status A patent was submitted
B ACKGROUND AND T E C HN O L O G Y
Psoriasis is a very common chronic skin disorder characterized as dermal inflammation that varies in severity, from minor, localized patches to complete body coverage. It affects ~2–3% of the population, making it one of the most prevalent autoimmune diseases worldwide. Psoriatic patients who suffer from moderate to severe disease, report that this has a significant negative impact on their quality of life. According to the National Psoriasis Foundation, USA, the total annual direct and indirect costs of psoriasis are over 11 billion US$. Despite the accessibility, frequency and chronicity of this complex disease, the fact that it involves several cell types and many genes makes many puzzling questions unanswered. This multigenic disease is characterized by abnormally increased keratinocytes (KC) proliferation, abnormal differentiation of the epidermis and systemic and local inflammation. These result in the formation of chronic erythematous, scaly lesions that are usually distributed symmetrically on the skin. Recently, it has been shown that a discrete population of lymphocytes, Th17 cells, were significantly more abundant in psoriatic than in normal skin. Th17 produce IL-17A, IL-17F, TNF-, IL-21 and IL-22.
Activated TH1, TH17- and NK-cells are the main sources of IL-22. IL-17A acts via heterodimeric receptor complex consisting of IL-17RA and IL-17RC. IL-22 acts via a heterodimeric receptor complex consisting of IL-22RA1 and IL-10RB. Neither resting nor activated immune cells express IL-22 receptor or respond to IL-22. In contrast, cells located in outer body barriers, i.e. skin, kidney the digestive and respiratory systems, are targets of this cytokine. Both IL-17 and IL-22 play a major role in the pathogenesis of psoriasis. Psoriatic patients display markedly elevated IL-22 plasma levels, which correlate with the disease severity. In an in vitro reconstituted human epidermal system, IL-22 triggered keratinocytes hyperplasia, leading to increased overall thickness of the epidermis. Moreover, neutralization of IL-22 prevents the development of psoriasis in a SCID mouse model for this disease. These findings form the basis for the utilization of inhibitors of the IL-17 and IL-22 pathways as possible therapeutic means in psoriasis.
Psoriasis treatments: About 80% of psoriatic patients are considered to have a mild disease and therefore may be adequately treated using topical therapies. Patients characterized as moderate-to-severe psoriasis generally require treatment with traditional systemic or new biologic therapies. Despite this fact most newly developed anti psoriatic drugs are for systemic treatments. As written above, the evolution of psoriasis involves interplay of KC and immunocytes and most drugs are targeted at one specific molecule and not at molecular networks. For example, infliximab, etanercept, adalimumab and golimumab are anti- is a monoclonal antibody to IL-12p40. Briakinumab (AMG-827, Amgen) is a new drug, now in Phase II studies, which is a fully human monoclonal antibody against IL-17 receptor A. Fezakinumab (ILV-094, Pfizer) is a fully human monoclonal IgGy antibody towards IL-22, in phase I study. Interestingly miRNAs can target more than one molecule and might target a whole molecular network. Therefore, miRNAs may have advantage as therapeutic means. תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 MiRNAs are 22nt non coding RNAs that can suppress the expression of protein-coding genes by targeting cognate messenger RNAs for translational repression or degradation. In the human genome, miRNAs comprise 1-5% of all genes. Computational predictions suggest that a single miRNA can target hundreds of mRNAs and that a single mRNA can be under the regulatory control of multiple miRNAs. As a consequence, miRNAs play important roles in the regulation of essential cellular activities such as proliferation, morphogenesis, apoptosis and differentiation.
MiRNAs in normal and psoriatic skin: The role of miRNAs in skin development was demonstrated recently in a mouse model constructed with conditional knockout of Dicer in the skin. It was found that miRNAs are essential for skin development. Yi et al. found more than 100 miRNAs in the skin and showed that the epidermis and hair follicles express discrete miRNA families. Loss of KC-specific Dicer expression produces several distinct defects in the skin that affect both the epithelium and epithelial- mesenchymal signaling in addition to hyper proliferation and decreased apoptosis. Hence, the importance of miRNAs in skin development has been revealed.
We compared miRNA expression in healthy skin to psoriatic involved and uninvolved skin using microarray technology. We found that in the psoriatic lesion the expression of several miRNAs differs from the normal skin. One of the miRNAs which differ between normal and psoriatic involved skin is mir-197. MiR-197 expression is down regulated in psoriatic lesions. We found that miR-197 can affect KC differentiation, proliferation and migration. It inhibits the expression of IL17RA and IL22RA1 and thus it inhibits both IL-17 and IL-22 signaling. Thus, miR-197 can affect several targets which play a major role in the pathogenesis of psoriasis and in the interaction between immunocytes and KC. These findings make miR-197 an attractive therapeutic molecule for psoriasis
T H E N EED Psoriasis is a non-curable chronic inflammatory skin disease which affects approximately 3% of the world population (Estimated >125 million psoriasis sufferers worldwide). Psoriasis has a major impact on the quality of life, leading psoriatic patients to report on reduction in physical and mental well being comparable with that seen in cancer, arthritis, hypertension, heart disease, diabetes, and depression. Thus, a safe and efficient cure is in immediate need. Most of psoriatic patients are considered to suffer from a mild disease and be adequately treated using topical therapies, however most newly developed anti psoriatic drugs are for systemic treatments. The possible use of miRNAs, as topical therapy in psoriasis, is very attractive regarding the ease of selective application and considering the enhanced permeability of the inflamed psoriatic skin. In order to validate the feasibility of miRNAs as a safe and effective therapeutic tool for psoriasis, a mouse model of human psoriatic skin is required in order to gain durable IP before phase I clinical trials.
P O T E N T I AL A PPLICATIONS Our main hypothesis is that changes in miRNAs expression in psoriatic skin may play a major role in its pathogenesis. Our model system allows us to understand whether miRNAs are involved in the pathogenesis of psoriasis. These findings may be of utmost importance in the understanding of the pathogenesis of this complex and common disorder. The discovery of novel molecular targets may open the way towards development of RNA based drugs delivered by topical or systemic ways. Currently we have discovered two such miRNA which can be developed as pharmaceutical compositions to treat psoriasis and cancer diseases in human
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A DVANTAGES Topical miRNA based therapy of psoriasis, may prove to be a safe, highly specific therapy, lacking systemic side effects. It may replace the corticosteroid based topical preparations, with the advantage of no systemic absorption.
D EVELOPMENT S T A G E We explored the miRNAs expression profile in normal versus psoriatic lesion or versus un-involved psoriatic skin. Our analysis revealed 14 novel miRNAs that were expressed differentially between psoriatic skin samples and normal skin biopsies. One of those miRNAs was miR-197, qRT-PCR as well as ISH indicate that it is expressed in keratinocytes and down regulated by more than three folds in psoriatic lesions compared to normal or uninvolved psoriatic skin. By constructing a HaCaT, human keratinocytes cell line, which stably over expresses miR-197 or pMSCV-HTR as a control we studied the role of miR-197 in keratinocytes biology. From these experiments our results indicate that miR-197 slows keratinocytes proliferation rate and directs them towards differentiation.
One of the main challenges of miRNAs research is finding their direct biochemical target. Bioinformatics analysis predicted that mir-197 targets the two units of the hetrodimeric IL-22R, namely IL22RA1 and IL10RB (9, 28). We found that that only IL22RA1 is a bona-fide target of miR-197 in a luciferase- reporter assay as well as WB assay. The data that miR-197 targeted both the IL22RA1 subunit of the IL- 22 receptor and the IL17RA receptor subunit emphasizes and reinforces the role which miR-197 might play in the pathogenesis of psoriasis. IL22RA1 subunit is part of the receptor of both IL-22 and IL-20 (9). The two of them are up regulated in psoriasis and have similar biological effects ((29) and references therein). While IL-22, utilizes only the IL22RA1/ IL10RB receptor, IL-20 can utilize also the IL20RA1/ IL20RA2 receptor (9). However, in psoriasis only the IL22RA1 subunit is up regulated (30). This implies that miR-197, by controlling the IL17RA and IL22RA1, can tune several aspects of this signaling cycle which is highly expressed in psoriasis.
We monitored the expression of miR-197 in PHK cells 1 and 48 h after treatment with different concentrations of IL-22 or IL-17 or both. The expression of miR-197 increased significantly in cells treated with either IL-17, or IL-22, or both for 1or 48h, as compared to untreated cells (p=0.022) (1h- Fig 4, 48h-data not shown). These results indicate that both IL17 and IL-22 signaling enhances the expression of miR-197.
IL-22 primarily activates the JAK/STAT pathway following IL-22 receptor activation. IL-22-induced signal transduction is dependent on tyrosine phosphorylation of STAT3 (review in (9) and references there in). We show that miR-197 activation by IL-22 is mediated through STAT3, since in the presence of STAT3 specific inhibitor; miR-197 is no longer induced in response to IL-22 treatment. MiR-197, which is located on human chromosome 1p13.3, is intergenic and therefore its expression is regulated by its own promoter. We have identified, using bioinformatics tools, that the putative promoter of miR-197 may contain several STAT binding sites; (TTN(4–6)AA) (31, 32) (Fig. 6A). Indeed, we were able to show, by Chromatin Immunoprecipitation (ChIP) assay that activated STAT3 binds to the miR-197 putative promoter region upon IL-22 treatment. In parallel we have results showing that IL-17 activated the expression of miR-197 through C/EBP binding to miR-197 promoter. This result indicates that upon treatment of PHK with IL-22, activated STAT3 binds to the miR-197 promoter region to form a feedback inhibition loop between IL-22, p-STAT3 and miR-197 which down regulates IL-22 receptor IL22RA1 subunit. The same is correct for IL-17, upon treatment of PHK with IL-17 there is activation of miR-197 which down regulates IL-17receptor IL17RA subunit.
It was previously shown that IL-22 enhances keratinocytes proliferation, down-regulates filaggrin, a keratinocytes differentiation marker (16, 33) and enhances keratinocytes migration through the activation תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 of mobility- and migration-regulating proteins (15, 34, 35). We asked whether these biological effects of IL-22 will be affected by miR-197. To address this question we treated over express miR-197 HaCaT cells with IL-22 and performed a BrdU incorporation assay as well as Migration Assay. HTR-HaCaT migration ability was 40% without IL-22 and acceded to 56% after IL-22 treatment. This motility was almost abolished in HaCaT cells over expressing miR-197; their motility was only 5-10%. Meaning miR- 197 overexpression significantly reduces the effect of IL-22 on keratinocytes.
In the case of IL-17 it was shown that IL-17 increased the expression of CC chemokine ligand (CCL20). We found that in cells over expressing miR-197 the activation of CCL20 in PHK treated with IL-17 is inhibited.
The present study determines that miR-197 is a regulator of both IL-22, IL22RA1, JAK/STAT3 signaling pathway, and the IL-17, IL17RA, which are known to play major role in the pathogenesis of psoriasis. Moreover, miR-197 abolished proliferation and migration of keratinocytes induced by IL-22. The function of individual miRNAs may be cell-type specific. IL22RA1 expression is restricted to pancreas, kidney and skin. This may imply that in these tissues, similar to keratinocytes, the main role of miR-197 might be the regulation of cellular response to IL-22 in a concentration dependent manner . In summery- we have identified a feedback mechanism controlling IL-17 and IL-22 pathways and miR- 197 expression, which sheds new light on the regulation of the interaction between inflammatory mediators and keratinocytes. The regulation of this signaling pathway may be important in inflammatory skin disorders and in wound healing. From our results miR-197 seems attractive therapeutic tool as it target both the proliferation of keratinocytes and can control two major signaling of IL-17 and IL- 22 which play a role in the interaction of the immune cells with the keratinocytes especially in psoriasis.
Recently we establishment a psoriatic mouse model for human psoriasis in Sheba: Our aim is to generate miRNA base therapy for psoriasis. Our initial miRNAs screening was from human biopsies. However, most of our understanding on the biology of miRNAs in KC was obtained from tissue culture experiments. To better understand the role of miRNAs in psoriasis and to open the way for therapeutic application of these findings, in vivo research must be done. There is at least one animal model for human psoriasis, which has been established and validated. This model is technicaly demanding and we established it in Sheba Medical Center. In this model, normal human skin is engrafted onto SCID mice. The mice are injected by natural killer cells (NK) cells obtained from psoriatic patients or autologous lymphocytes activated by bacterial super antigens. The human skin develops into a psoriatiform lesion. This model is well established and is recognized as a mouse model for human psoriasis. Delivering naked miRNA is problematic due to its enzymatic degradation in vitro or in vivo. Furthermore, its cellular uptake is poor since miRNAs, as well as cells’ membranes, are negatively charged. Thus, miRNA delivery system needs to be considered in order to establish an efficient psoriatic treatment. In addition, since the skin is permeable to molecules smaller than 0.5 kDa, delivery of miRNA (14kDa) to skin cells requires methods which enhance transdermal delivery. We established collaboration with Professor Kost J. from the Department of Chemical Engineering at the Ben Gurion University. Professor Kost developed technique of sonication (US) of the skin in order to change the penetrability of the skin allowing delivery of small RNAs using a specified vehicle to the epidermis. We tried applied US on normal human skin that engrafted onto SCID mice, and in preliminary results we found that the labeled miR-197 penetrated the epiderimes of the human skin.
Moreover, we found that the penetrated miR-197 is biochemically active, as it down regulated the expression of its target, IL22RA1. In further experiments we applied the miRNA on the psoriatic lesion. A week after a single application we measured by qRT-PCR the amount of miR-197 in the lesion of treated mice compared to untreated mice and it was about 100 fold more in the treated mice. We next analyzed the biological effect of the treatment. In the next experiment the psoriasis model was generated in 19 mice which were divided into 4 groups; 1) injected by PBS and treated by mir-197. 2) Injected by activated lymphocytes and treated by scrambled miRNA. 3) Injected by activated psoriatic T cells without further treatment. 4) Injected by activated psoriatic T cells and treated by miR-197 as a single exposure. All groups were sacrificed after a week; biopsies were taken from the implanted skins of תל השומר מחקרים תשתיות ושירותים רפואיים Tel Hashomer Medical Research, Infrastructure and Services Tel: +972-3-5305998 Fax: +972-3-5305944 all mice and Formalin-fixed, paraffin-embedded (FFPE) blocks were prepared and stained with H&E. Each biopsy was analyzed in an absolutely blinded manner by a derma-pathologist and was evaluated by the use of five criteria that are serve as measurement for the severity of psoriasis; (1) the epidermal thickness (2) the uniform shape of the epidermis (3) the magnitude of the parakeratosis (the retention of nuclei in the stratum corneum) (4) the presence of neutrophils in the dermis and epidermis (5) the number of blood vessels. We found that miR-197 treatment improved significantly the psoriatic features compared to the untreated skin or scrambled RNA treated skin.
P A T E N T Pending
T H E N E E D Safe and effective topical therapy for psoriasis that may be used for the long run without significant side effects is still lacking. Topical miRNA based therapy may provide this hope.
T H E M ARKET Psoriasis affects 2–3% of the population, making it one of the most prevalent autoimmune diseases worldwide, and can be associated with other inflammatory conditions, such as psoriatic arthritis, inflammatory bowel disease and coronary artery disease. 75% of the psoriatic patients has mild to moderate disease - among them the majority ~95% are treated only by topical therapeutics and phototherapy. The estimated market value of topical agents in psoriasis is $850 million, and the market for biologics is approaching $2.5 billion (1).
This market is expected to grow significantly in the future, largely due to the chronic nature of the disease and the high unmet need. The uptake of biologics, more therapies emerging, and the continued need for topical therapies will make this market approachable for any new therapy that is effective and safe.
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