Bessor Pharma

RENALASE TECHNOLOGY

RENALASE TECHNOLOGY

Renalase (RNLS) is a secreted flavoprotein, produced in the kidney, pancreas and other tissues, that was discovered by our collaborator, Dr. Gary Desir, Chairman of Medicine at Yale and colleagues. Dr. Desir’s group has shown that RNLS can function as a survival factor that acts through a pro-survival, anti-apoptotic signaling cascade (MAPK). This work has established a strong therapeutic rationale for the short-term activation of the RNLS signaling pathway with RNLS peptide that mimics RNLS to treat a number of acute conditions characterized by RNLS deficiency, including pancreatitis, acute kidney injury, and protection against the kidney-toxic effects of chemotherapeutics such as cisplatin.

In addition, they have shown that RNLS is a promising new target for pancreatic cancer, melanoma, and potentially other cancers. Both projects are on a path to an IND in approximately 18 months. RNLS levels’ link to these conditions provides a potential new and useful biomarker for early diagnosis and monitoring of drug response.

Bessor has exclusive rights from Yale three issued patents and to patent applications covering RNLS, analogs, regulators, and diagnostic assays. We have established an LLC, Personal Therapeutics to translate these discoveries into therapeutic candidates.

Renalase Agonists

ACUTE PANCREATITIS AND ACUTE KIDNEY INJURY

We are developing a peptide agonist of the secreted flavoprotein renalase (RNLS) to prevent and treat acute pancreatitis (AP) and acute kidney injury (AKI), with RNLS as a biomarker and diagnostic for early detection. We have established a rationale for RNLS agonists in AP and AKI based on data demonstrating:

  • RNLS is produced in the kidneys and pancreas and the severity of AKI or AP in mice increases with falls in RNLS levels.
  • Serum RNLS levels are decreased in mice and humans with AP and AKI
  • Exogenous RNLS reduces tissue injury in models of AP and AKI.
  • In addition to its enzymatic activity in metabolizing catecholamines, RNLS can function as a survival factor that acts through a membrane receptor to initiate a pro-survival, anti-apoptotic signaling cascade (MAPK).
  • We designed and synthesized small peptides to mimic RNLS regions that interact with its receptor and protects against AKI and AP have shown proof-of-concept (POC)

Target Markets
Acute pancreatitis is the most frequent gastrointestinal cause for hospital admission in the US with increasing incidence and is lethal in up to 30 percent of severe cases. AP has multiple causes but inflammation, pain, and cell death are universal hallmarks. Inflammation and edema in the kidney, and particularly the lung as a result of pancreatitis, are major contributors to pancreatitis lethality.

Acute kidney injury is a clinical condition commonly associated with sepsis, surgery, and certain drugs, and which affects up to 20 percent of hospitalized patients. Epidemiologic data indicate the severity of AKI is associated with in-hospital and follow on mortality as well as enormous healthcare costs.

Innovations and Potential Advantages
There are few effective therapeutic options for AP other than supportive care and none are approved in the US.

Other than supportive care there are no approved therapies for AKI. New therapeutic development has been hampered by lack of a predictive biomarker that can rapidly identify patients at risk of AKI. Current diagnosis relies heavily on serum creatinine measurements, and is therefore delayed by a critical 48-72 hours. Our biomarker-driven approach has potential to address this issue.

Status
We have demonstrated in vivo POC in models of AP and AKI with RNLS and candidate peptides designed to mimic RNLS’ binding with its receptor. We are also developing an improved and validated assay to better detect RNLS in plasma. Bessor, which has collaborated with the Desir team for more than 24 months, including with Dr. Fred Gorelick (pancreatitis), has exclusive rights from Yale for RNLS, analogs and RNLS diagnostic assays. The AP project has been validated through support of an SBIR grant from the National Institute of Diabetes, Digestive and Kidney Diseases. Broad patent applications are filed. RNLS and peptide supplies are being scaled up for further preclinical and toxicology studies with the goal to file an IND in approximately 18 months.

Publications
Renalase Protects Against Acute Pancreatitis. Pancreas. Nov. 2014, V.43 – Issue 8 – p 1340–1429

Renalase prevents AKI independent of amine oxidase activity. J Am Soc Nephrol. 2014 Jun;25(6):1226-35.

PROTECTION AGAINST CHEMOTHERAPEUTIC-INDUCED KIDNEY INJURY

Too often, otherwise effective chemotherapy, such as cisplatin, must be stopped due to drug induced kidney toxicity because there are no effective drugs to blunt such serious adverse effects. We are developing renalase peptide agonists with the potential to allow longer use of effective anticancer therapies by treating acute chemotherapy-induced kidney injury and preventing progression to chronic kidney disease. We have established in vivo proof of concept for RNLS agonist use in cisplatin mediated kidney toxicity based on data demonstrating:

  •  In man, RNLS deficiency is associated with dramatically more severe cisplatin-mediated acute kidney injury, elevated plasma creatinine, and chronic kidney injury.
  • Acute administration of RNLS protein ameliorates cisplatin-induced toxicity in in vivo preclinical models.
  • RNLS can function as a survival factor that acts through a membrane receptor to initiate a pro-survival, anti-apoptotic signaling cascade (MAPK).
  • We have now designed, produced, and shown that our proprietary peptides mimic RNLS regions that interact with its receptor and protect against cisplatin renal toxicity. These peptides are being developed as therapeutic candidates.

In addition, RNLS’ association with kidney toxicity suggests that RNLS plasma levels may be a potential biomarker for otherwise non obvious emerging cisplatin induced kidney toxicity, and a marker for drug response. There is a strong group advancing this project which is a collaboration with Dr. Gary Desir, Chairman of Medicine at Yale, and his team.

Target Markets
The initial target market is patients undergoing cancer treatment with cisplatin, one of the most widely used chemotherapies. It is a named component in half of the drug cocktails used for cancer. Cisplatin is approved for treating many cancers including bladder, cervical, non-small cell lung, ovarian, testicular cancer, malignant mesothelioma, and squamous cell carcinoma of the head and neck. Kidney toxicity severely limits use of cisplatin, with 25-35% of patients required to reduce dosing or stop otherwise effective therapy. Follow-on indications include renal toxicities of other platinum-based drug and immuno-inflammation linked toxicities of other chemotherapeutic drugs including carboplatin, nitrosoureas, and methotrexate.

Innovations and Potential Advantages
There is no widely used effective therapy to reduce cisplatin induced kidney toxicity. There is one approved compound for use in ovarian cancer, amifostine, which carries a black box warning and is not widely used.

To help guide therapeutic development, we have also developed improved models of AKI that reproduce the AKI and chronic kidney disease pattern observed in humans treated with cisplatin.

The therapeutic we are developing is a peptide, which can be readily manufactured cost effectively and may have other therapeutic uses.

The project includes the potential for biomarker development for early kidney toxicity detection, and a marker for drug response.

Status
We have demonstrated initial in vivo proof-of-concept. In single dose experiments candidate peptides and RNLS prevent and treat cisplatin-induced AKI in mice by reducing renal necrosis, apoptosis, and inflammation, and by blunting 50% of the acute rise in plasma creatinine. Adding RNLS or agonist for 24-72 hours neither diminishes cisplatin effectiveness against cancer cells, nor promotes cancer cell growth.

The project has been supported by a Phase 1 STTR grant from the National Cancer Institute. We have a lead compound and back up poised for further development towards IND.

We are on a course to select and characterize a lead candidate peptide agonist in vitro and in vivo to further define its efficacy, safety, and non-interference with cisplatin anti-tumor activity; to develop an ELISA for detecting the lead in plasma, and to produce GMP material for IND directed studies. There is rationale and evidence that the mechanism involved with the renal protective effects of RNLS agonists may prevent serious tissue injury in other organ systems caused by platinum-based drugs and significantly improve the therapeutic index of other chemotherapy agents.

Intellectual Property
Bessor Pharma and its experienced R&D team including cancer therapy experts have been collaborating with the Desir team for more than 24 months to advance this project. Bessor has exclusive rights from Yale to three issued renalase patents and pending applications for renalase, analogs, renalase regulators and renalase diagnostic assays. Broad patent applications are filed.

Publications
Three-Dimensional Morphology by Multiphoton Microscopy with Clearing in a Model of Cisplatin-Induced CKD. J Am Soc Nephrol. 2016 Apr;27(4):1102-12.

Renalase protects against cisplatin acute kidney injury in mice. The FASEB Journal April 2013 vol. 27 No. 1 Supplement 910.7

Anti-Renalase Antibodies

PANCREATIC CANCER AND MELANOMA

Renalase has recently emerged as a promising target for pancreatic cancer and melanoma, based on work by our collaborator, Dr. Gary Desir, Chairman of Medicine at Yale and a pioneer in renalase biology. Several lines of evidence underscore (1) RNLS’ potential as an important and innovative target and biomarker in melanoma and pancreatic cancer; and (2) a unique potential therapeutic role for a RNLS mAb.

  • Mechanisms shown. RNLS functions include those of survival factor that acts through a membrane receptor to initiate a pro-survival, anti-apoptotic signaling cascade.
  • Proof of mechanism with clinical correlates. Expression is elevated in pancreatic tumors, melanoma, and other cancers (e.g. bladder). RNLS expression levels inversely correlate with survival in melanoma and pancreatic cancer patients;
  • In vivo and in vitro proof-of-concept (POC) established. Signaling inhibition has anti-tumor activity in vitro (shRNA and anti-RNLS antibodies) and in vivo in xenograft models of melanoma and pancreatic cancer (anti-RNLS antibodies);
  • Desired effects on tumor micro- environment and macrophage polarization and RNLS Ab active in cancers resistant to current therapy;
  • Strong IP, top R&D team and know-how; preclinical stage, expected 18 months to IND

RNLS was initially identified as hormone produced by the kidney that degrades circulating catacholamines. RNLS’ pro-survival activity is independent of its enzymatic activity, with evidence consistent with cancers “hijacking” and overexpressing RNLS to thwart host defense. Evidence to date indicates a RNLS mAb therapeutic anticancer drug would have a more than acceptable therapeutic index.

Initial Target Markets
Melanoma
The incidence of melanoma is not only rising faster than that of any other malignancy, but is also coupled with a dramatic increase in the incidence of metastatic disease and death. Despite recent significant advances, such as immune checkpoint inhibitors, many patients are not cured and the projected death rate from melanoma in 2015 in the United States has risen by 30% to 9,940 from 7,600 in 2013.

Pancreatic Cancer
Pancreatic ductal adenocarcinoma (PDAC) is among the deadliest cancers. It is the 9th most common tumor type, but ranks 3rd in cancer deaths, with an estimated 41,780 in the US, according to the American Cancer Society. PDAC is among the few cancer types for which survival has not markedly improved in more than 40 years. Current drug therapy is minimally effective.

Innovations and Potential Advantages
We have shown that RNLS’ activity as a survival factor is based on a novel mechanism. RNLS activates specific signaling pathways (MAPK, PI3K and JAK/STAT3) that protect cells against apoptosis. The receptor for extracellular RNLS and the key RNLS region that binds to it have been identified. The candidates identified to include date include mABs that target the binding region.

Our collaborators at the Desir lab at Yale have demonstrated that RNLS is preferentially expressed in CD163+ (M2-like) tumor associated macrophages (TAMs); facilitates melanoma growth by activating STAT3 in tumor cells and TAMs; and treatment of tumor bearing animals with an inhibitory anti-renalase monoclonal antibody led to a marked reduction in CD163+ TAMs. Hypoxia inducing factor upregulates RNLS.

RNLS’ putative role in cancer and potential as a drug target also suggests promise as marker for patient selection and drug response.

Status
We have established a therapeutic rationale for an anti-renalase mAb and demonstrated preclinical in vivo proof of concept in melanoma and PDAC. We are humanizing the lead RNLS mAb and believe we can be at IND in 18 months.

Melanoma
We have shown that RNLS expression in melanoma cells is higher in metastatic than primary tumors. Knock-down of RNLS by siRNA decreases melanoma cell survival in vitro. This finding is confirmed by anti-RNLS antibodies and an inhibitory peptide in vitro and using xenograft murine models. Moreover, RNLS expression in human tumors was also seen in CD163+ TAMs. Treatment of tumor bearing animals with an inhibitory anti-renalase monoclonal antibody led to a marked reduction in CD163+ TAMs. We also showed that increased RNLS production by CD163+ TAMs facilitates melanoma growth by activating STAT3 in tumor cells and TAMs, suggesting that RNLS inhibition of both melanocytes and TAMs might be a useful approach for treating melanoma.

Pancreatic Ductal Adenocarcinoma
RNLS is expressed in PDAC cells and RNLS expression in humans with PDAC inversely correlates with patient survival. In a cohort of individuals with PDAC, those whose tumors expressed high RNLS levels (n=34) had reduced 3-year survival rate vs. those with low expression (n=34; 24% versus 49%, p=0.024). We have also determined that RNLS inhibition using shRNA technology reduced pancreatic cancer cell growth by >90% in vitro, and decreased tumor volume by 80% in mouse xenograft models (animals sacrificed at day 30 post treatment).

We have identified several candidate antibodies (rabbit) that inhibit pancreatic cancer cell growth in vitro. One antibody has been tested in vivo, in a xenograft model, resulting in a 78% decrease in PDAC tumor volume (animals sacrificed at day 24 post treatment). Examination of the tumor samples showed that the mAb caused a marked reduction in RNLS expression and STAT3 activation, and increased the rate of apoptosis.

Future Directions
Scale up supplies of RNLS and therapeutic antibody are being produced for the necessary further preclinical and toxicology studies with the goal to file an IND in about 18 months. Development of an advanced RNLS assay is underway. The availability of an assay for RNLS and therapeutic antibodies already developed in our initial project can be very efficiently utilized for utility and future development in other cancers where RNLS levels are high (> 100 fold in certain cancers), including breast, bladder, neuroendocrine cancers, and endometrial cancer.

Intellectual Property and Strong R&D Team
Bessor Pharma’s experienced R&D team and advisors (including Dr. James Abbruzzese, Duke) along with Yale collaborators (including Drs. Desir and Harriet Kluger) as well as other cancer therapy experts have been collaborating for more than 24 months to advance this project. Bessor has exclusive rights from Yale to three issued renalase patents and applications for RNLS, analogs, RNLS regulators and antibodies and RNLS diagnostic assays. Broad patent applications are filed.

Publications
Renalase expression by melanoma and tumor associated-macrophages promotes tumor growth through a STAT3-mediated mechanism. Cancer Res. 2016 May 9.

Inhibition of renalase expression and signaling has antitumor activity in pancreatic cancer. Sci Rep. 2016 Mar 14;6:22996.

Pancreatic Ductal Adenocarcinoma: From Genetics to Biology to Radiobiology to Oncoimmunology and All the Way Back to the Clinic Biochim Biophys Acta. 2014 Dec 6.

Renalase prevents AKI independent of amine oxidase activity. J Am Soc Nephrol. 2014 Jun;25(6):1226-35.

The early detection of pancreatic cancer: what will it take to diagnose and treat curable pancreatic neoplasia? Cancer Res. 2014 Jul 1;74(13):3381-9.