Lake Fjord Pass

DEVELOPMENT OF A PROTEIN DRUG FOR PANCREATIC CANCER TREATMENT - ABSTRACT PANCREATIC CANCERS ARE DEVASTATING DISEASES WITH FIVE YEAR SURVIVAL RATE LESS THAN 9%. CURRENTLY, THERE IS NO EFFECTIVE TREATMENT FOR ADVANCED DISEASE. ONE MAJOR BARRIER TO EFFICACY OF ANTI-TUMOR THERAPEUTICS IS THE DENSE DESMOPLASTIC STROMAL RESPONSE. EVIDENCE SUGGESTS THAT CANCER ASSOCIATED PANCREATIC STELLATE CELLS (CAPSC) PRODUCE THE STROMAL COLLAGEN. THE ECM LAID DOWN BY CAPSC IS CONSIDERED TO BE ONE OF THE MAJOR CONTRIBUTORS OF RESISTANCE TO ESTABLISHED THERAPIES OF THE DISEASES. DEPLETING CAPSC AND ALTERING VESSEL DENSITY COULD SIGNIFICANTLY IMPROVE EFFICACY OF EXISTING PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) TREATMENTS. HOWEVER, CURRENTLY, THERE ARE NO APPROVED THERAPIES THAT ARE ABLE TO DEPLETE CAPSCS IN PDAC. WE HAVE DEVELOPED A NOVEL THERAPEUTIC PROTEIN (PROAGIO) USING RATIONAL PROTEIN DESIGN. PROAGIO IS DESIGNED TO TARGET INTEGRIN ΑVΒ3 AT A NOVEL SITE (NOT THE LIGAND BINDING SITE). PROAGIO SPECIFICALLY INDUCES APOPTOSIS OF INTEGRIN ΑVΒ3 EXPRESSING CELLS WITH HIGH EFFICACY BY A NOVEL MECHANISM OF DRUG ACTION (RECRUITING & ACTIVATING CASPASE 8 AT CYTOPLASMIC DOMAIN OF Β3). WE REASONED THAT, SINCE BOTH CAPSC AND ANGIOGENIC ENDOTHELIAL CELLS EXPRESS HIGH LEVELS OF INTEGRIN ΑVΒ3, AND SINCE PROAGIO IS VERY EFFECTIVE IN INDUCING APOPTOSIS OF INTEGRIN ΑVΒ3 EXPRESSING CELLS, PROAGIO SHOULD BOTH DEPLETE CAPSC AND ELIMINATE NEW BLOOD VESSELS IN AND AROUND PANCREATIC TUMORS. THIS UNIQUE STRATEGY MAY PROVE ADVANTAGEOUS IN TREATMENT OF PDAC. OUR STTR PHASE I&II STUDIES DEMONSTRATED EFFICACY OF PROAGIO POTENTIALLY AS A PDAC TREATMENT VIA VARIOUS CANCER MODELS. THE STUDIES SUPPORT OUR HYPOTHESIS THAT PROAGIO CAN PROVIDE TREATMENT BENEFIT BY SIMULTANEOUSLY DEPLETING THE COLLAGEN-PRODUCING CAPSCS THAT SUPPORT TUMOR DESMOPLASIA AND CANCER CELL GROWTH, WHILE ALSO ELIMINATING NEWLY GROWN CANCER ASSOCIATED BLOOD VESSELS THAT FEED CANCER CELLS AND ENABLE CANCER METASTASIS. DATA FROM OUR STTR PHASE I&II STUDIES PROVIDES PROOF OF PRINCIPLE FOR FUTURE CLINICAL TESTS. RESULTS FROM OUR PHASE II STUDIES HAVE LED TO IND APPLICATION OF PROAGIO AS A PANCREATIC CANCER TREATMENT DRUG. THE MAIN OBJECTIVE OF THIS PHASE IIB APPLICATION IS TO GENERATE A DEFINITIVE DATASET TO ENABLE THE DEVELOPMENT OF PROAGIO AS A VIABLE THERAPEUTIC OPTION FOR PDAC PATIENTS. AIM 1 WILL CHARACTERIZE THE TOXICITY AND TOLERABILITY AND DETERMINE THE MAXIMUM TOLERATED DOSE (MTD) AND RECOMMENDED PHASE II DOSE (RP2D) OF PROAGIO AS A SINGLE AGENT AND IN COMBINATION WITH G-NP. AIM 2 WILL CHARACTERIZE PK PROPERTY OF PROAGIO IN CANCER PATIENTS AND TO OBTAIN PRELIMINARY ANTI-CANCER ACTIVITY DATA OF PROAGIO AND PROAGIO + G-NP IN PDAC PATIENTS. AIM 3 WILL ANALYZE THE EFFECTS OF PROAGIO IN PATIENT TUMOR TO VALIDATE THE MECHANISM OF DRUG ACTION IN PATIENTS. THIS STUDY WILL EXPLORE NEW THERAPEUTIC AVENUE FOR PDAC PATIENTS.

PRECISION MRI WITH PROTEIN CONTRAST AGENT FOR NONINVASIVE DETECTION OF LUNG FIBROSIS - SUMMARY LUNG DISEASES, SUCH AS INTERSTITIAL LUNG DISEASES, INCLUDING IDIOPATHIC PULMONARY FIBROSIS (IPF), CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), AND ACUTE VIRAL INFECTION, ARE MAJOR LEADING CAUSES OF DEATH WORLDWIDE. IPF IS CHARACTERIZED BY INCREASED EXTRACELLULAR MATRIX (ECM) DEPOSITION, ESPECIALLY TYPE I COLLAGEN, WITHIN THE LUNG, CAUSING IRREVERSIBLE LUNG DAMAGE. WHILE HIGH-RESOLUTION COMPUTED TOMOGRAPHY (HRCT) IS COMMONLY USED TO DIAGNOSE IPF, THIS METHOD CAN PRODUCE LARGE INTER-OBSERVER VARIATION AND ERROR (UP TO 50%) AND CANNOT DETECT EARLY-STAGE LUNG FIBROSIS DUE TO POOR SOFT TISSUE CONTRAST. A SURGICAL LUNG BIOPSY IS OFTEN REQUIRED TO CONFIRM THE DIAGNOSIS FOR PATIENTS WITH SUSPECTED IPF. HOWEVER, THIS IS VERY RISKY DUE TO COMORBIDITIES AND PHYSICAL IMPAIRMENTS. HRCT AND OTHER AVAILABLE DIAGNOSTIC METHODS PROVIDE LITTLE INFORMATION ABOUT HETEROGENEOUS FIBROSIS RELATED TO DISEASE ACTIVITY, PROGRESSION, AND REGRESSION. THERE IS A PRESSING UNMET MEDICAL NEED TO DEVELOP NONINVASIVE IMAGING METHODOLOGIES AND CONTRAST AGENTS TO DETECT EARLY STAGES OF LUNG FIBROSIS, AND TO STAGE FIBROSIS SEVERITY AND HETEROGENEOUS EXPRESSION OF COLLAGEN. THE MAIN OBJECTIVES OF THIS PHASE II APPLICATION WILL PROVIDE NECESSARY RESULTS FOR TRANSITION TO CLINICAL APPLICATION FOR NON-INVASIVE LONGITUDINAL EARLY DIAGNOSIS AND STAGING OF LUNG FIBROSIS WITH DISEASE ACTIVITY BY PMRI. AIM 1 IS TO ANALYZE TOXICOLOGY AND METAL TOXICITY OF PROCA32.COLLAGEN IN HEALTHY AND LUNG FIBROSIS MICE. WE WILL COMPLETE SINGLE AND REPEATING DOSE STUDIES TO VERIFY THAT INL-101 IS NOT TOXIC USING ESTABLISHED IPF ANIMAL MODELS INCLUDING REVERSIBLE AND IRREVERSIBLE FIBROSIS MODELS. WE WILL ALSO DETERMINE MINIMAL EFFECTIVE DOSE OF INL-101 FOR DETERMINING LUNG FIBROSIS. AIM 2 IS TO EXECUTE THE DESIGNED TOX/PK SAFE PROFILE STUDIES FOR IND APPLICATION AND PREPARE HUMAN IMAGING. WE WILL OPTIMIZE INJECTION ROUTES FOR IN VIVO IMAGING FOR IND APPLICATION. THIS TRANSFORMATIVE PLATFORM AND PROPRIETARY METHODOLOGY WILL HAVE STRONG IMPACT IN IPF PATIENT STRATIFICATION FOR EFFECTIVE TREATMENT AND REDUCTION OF MORTALITY BY DISTINGUISHING IPF PATIENTS WHO ARE EXPERIENCING RAPID PROGRESSION FROM THOSE WHO ARE STABLE BASED ON FEATURES OF REVERSIBLE AND IRREVERSIBLE FIBROSIS.

DESIGN OF CALCIUM-SENSING RECEPTOR TARGETED THERA(G)NOSTIC AGENTS - SUMMARY CALCIUM-SENSING RECEPTOR (CASR), AS ONE OF THE FAMILY C G-PROTEIN-COUPLED RECEPTORS (GPCRS), LARGELY MODULATES EXTRACELLULAR CA2+ BY TRIGGERING MULTIPLE INTRACELLULAR SIGNALING PATHWAYS. CASR IS EXPRESSED IN THE KIDNEYS, PARATHYROID GLANDS, COLON, AND OTHER CELLS, REPRESENTING ITS DIFFERENTIAL YET KEY ROLES IN RESPONDING TO EXTRACELLULAR CALCIUM, AMINO ACIDS, AND METABOLITES, IN DIFFERENT ORGANS. NUMEROUS DISEASES HAVE BEEN ATTRIBUTED TO CALCIUM MISHANDLING (E.G.,NEPHROLITHIASIS) AND SECONDARY HYPERPARATHYROIDISM (HPT), WITH OVER 200 CASR MUTATIONS THAT CAN SIGNIFICANTLY ALTER CASR EXPRESSION. THERE IS A PRESSING UNMET MEDICAL NEED TO DEVELOP POSITIVE ALLOSTERIC MODULATORS FOR PKD AND OTHER DISEASES WITH CELLULAR AND ORGAN SPECIFICITY AND REDUCED LIVER TOXICITY. THE MAJOR BARRIERS TO DEVELOPING EFFECTIVE DRUGS ARE A LACK OF KNOWLEDGE OF TEMPO-SPATIAL QUANTITATIVE EXPRESSION OF CASR IN THE BODY AND ITS CHANGES DURING DISEASE PROGRESSION AND REGRESSION. WE RECENTLY DETERMINED THE FIRST CRYSTAL STRUCTURE OF THE HUMAN CASR EXTRACELLULAR DOMAIN (ECD) WITH A BOUND TRYPTOPHAN DERIVATIVE (TNCA) AT 2.1 Å, A POTENTIAL POSITIVE ALLOSTERIC MODULATOR. THIS LED US TO DESIGN AND SYNTHESIZE A DRUG CANDIDATE TNCA THAT HAS STRONG AGONIST ACTIVITY AND SPECIFICITY TO CASR. THE GOAL OF THIS PHASE I PROPOSAL IS TO ACQUIRE PROOF-OF-CONCEPT RESULTS FOR DESIGN AND GENERATION OF TNCA ANALOGS HAVING OPTIMAL DRUG ACTIVITY AND CELLULAR SPECIFICITY AS AGONISTS AND ANTAGONISTS. IN ADDITION, WE WILL ALSO DEVELOP CASR TARGETED PET AGENTS FOR RENAL IMAGING IN A PRECLINICAL PKHD1PCK RAT MODEL (STUDIES IN THIS MODEL LED TO CLINICAL TRIALS OF AN FDA-APPROVED PKD THERAPEUTIC). AIM 1 IS TO OPTIMIZE AND DETERMINE TNCA ANALOGS WITH BEST CASR DRUG ACTIVITY AS AGONISTS & ANTAGONISTS. AIM 2 IS TO PERFORM IN VIVO MAPPING OF CASR DISTRIBUTION IN PKD (PKHD1PCK) RAT MODEL BY MICROPET IMAGING. PHASE II WILL FURTHER VALIDATE DRUG EFFICACY OF TNCA ANALOGS, IMAGING CAPABILITY FOR CASR ASSOCIATED KIDNEY DISEASES USING ANIMAL MODELS, AND SAFETY PROFILE FOR AN EIND. SUCCESS IN OUR PROPOSED STUDIES WILL PROVIDE ESSENTIAL RESULTS FOR A NOVEL AVENUE IN THE DEVELOPMENT OF CASR SPECIFIC DRUG CANDIDATES FOR THE EFFECTIVE TREATMENT OF VARIOUS CASR ASSOCIATED HUMAN DISEASES USING NEWLY AVAILABLE STRUCTURES AND PATENTED METHODOLOGY. IN ADDITION, THE DEVELOPED PET IMAGING AGENTS WILL ENABLE US TO MAP CASR DISTRIBUTION AND HETEROGENEOUS EXPRESSION DURING PKD DISEASE PROGRESSION AND REGRESSION IN VIVO, AS A COMPANION DIAGNOSTIC FOR PATIENT RISK STRATIFICATION, AND FOR OUTCOME PREDICTION AND NOVEL DRUG DEVELOPMENT. WE ANTICIPATE THAT THE PROPOSED STUDIES WILL FOSTER THE DEVELOPMENT OF TRANSFORMATIVE NOVEL MARKERS OF THE DISEASE ACTIVITY THAT WILL ENHANCE CLINICAL CARE OF PKD AND OTHER KIDNEY DISEASES.

DEVELOPMENT OF A PROTEIN DRUG FOR PANCREATIC CANCER TREATMENT - ABSTRACT PANCREATIC CANCERS ARE DEVASTATING DISEASES WITH FIVE YEAR SURVIVAL RATE LESS THAN 9%. CURRENTLY, THERE IS NO EFFECTIVE TREATMENT FOR ADVANCED DISEASE. ONE MAJOR BARRIER TO EFFICACY OF ANTI-TUMOR THERAPEUTICS IS THE DENSE DESMOPLASTIC STROMAL RESPONSE. EVIDENCE SUGGESTS THAT CANCER ASSOCIATED PANCREATIC STELLATE CELLS (CAPSC) PRODUCE THE STROMAL COLLAGEN. THE ECM LAID DOWN BY CAPSC IS CONSIDERED TO BE ONE OF THE MAJOR CONTRIBUTORS OF RESISTANCE TO ESTABLISHED THERAPIES OF THE DISEASES. DEPLETING CAPSC AND ALTERING VESSEL DENSITY COULD SIGNIFICANTLY IMPROVE EFFICACY OF EXISTING PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) TREATMENTS. HOWEVER, CURRENTLY, THERE ARE NO APPROVED THERAPIES THAT ARE ABLE TO DEPLETE CAPSCS IN PDAC. WE HAVE DEVELOPED A NOVEL THERAPEUTIC PROTEIN (PROAGIO) USING RATIONAL PROTEIN DESIGN. PROAGIO IS DESIGNED TO TARGET INTEGRIN ΑVΒ3 AT A NOVEL SITE (NOT THE LIGAND BINDING SITE). PROAGIO SPECIFICALLY INDUCES APOPTOSIS OF INTEGRIN ΑVΒ3 EXPRESSING CELLS WITH HIGH EFFICACY BY A NOVEL MECHANISM OF DRUG ACTION (RECRUITING & ACTIVATING CASPASE 8 AT CYTOPLASMIC DOMAIN OF Β3). WE REASONED THAT, SINCE BOTH CAPSC AND ANGIOGENIC ENDOTHELIAL CELLS EXPRESS HIGH LEVELS OF INTEGRIN ΑVΒ3, AND SINCE PROAGIO IS VERY EFFECTIVE IN INDUCING APOPTOSIS OF INTEGRIN ΑVΒ3 EXPRESSING CELLS, PROAGIO SHOULD BOTH DEPLETE CAPSC AND ELIMINATE NEW BLOOD VESSELS IN AND AROUND PANCREATIC TUMORS. THIS UNIQUE STRATEGY MAY PROVE ADVANTAGEOUS IN TREATMENT OF PDAC. OUR STTR PHASE I&II STUDIES DEMONSTRATED EFFICACY OF PROAGIO POTENTIALLY AS A PDAC TREATMENT VIA VARIOUS CANCER MODELS. THE STUDIES SUPPORT OUR HYPOTHESIS THAT PROAGIO CAN PROVIDE TREATMENT BENEFIT BY SIMULTANEOUSLY DEPLETING THE COLLAGEN-PRODUCING CAPSCS THAT SUPPORT TUMOR DESMOPLASIA AND CANCER CELL GROWTH, WHILE ALSO ELIMINATING NEWLY GROWN CANCER ASSOCIATED BLOOD VESSELS THAT FEED CANCER CELLS AND ENABLE CANCER METASTASIS. DATA FROM OUR STTR PHASE I&II STUDIES PROVIDES PROOF OF PRINCIPLE FOR FUTURE CLINICAL TESTS. RESULTS FROM OUR PHASE II STUDIES HAVE LED TO IND APPLICATION OF PROAGIO AS A PANCREATIC CANCER TREATMENT DRUG. THE MAIN OBJECTIVE OF THIS PHASE IIB APPLICATION IS TO GENERATE A DEFINITIVE DATASET TO ENABLE THE DEVELOPMENT OF PROAGIO AS A VIABLE THERAPEUTIC OPTION FOR PDAC PATIENTS. AIM 1 WILL CHARACTERIZE THE TOXICITY AND TOLERABILITY AND DETERMINE THE MAXIMUM TOLERATED DOSE (MTD) AND RECOMMENDED PHASE II DOSE (RP2D) OF PROAGIO AS A SINGLE AGENT AND IN COMBINATION WITH G-NP. AIM 2 WILL CHARACTERIZE PK PROPERTY OF PROAGIO IN CANCER PATIENTS AND TO OBTAIN PRELIMINARY ANTI-CANCER ACTIVITY DATA OF PROAGIO AND PROAGIO + G-NP IN PDAC PATIENTS. AIM 3 WILL ANALYZE THE EFFECTS OF PROAGIO IN PATIENT TUMOR TO VALIDATE THE MECHANISM OF DRUG ACTION IN PATIENTS. THIS STUDY WILL EXPLORE NEW THERAPEUTIC AVENUE FOR PDAC PATIENTS.

NON-INVASIVE STAGING OF METASTASIS BY PRECISION MRI - PROJECT SUMMARY/ABSTRACT THIS SBIR APPLICATION WILL DEVELOP AN IMPROVED CONTRAST AGENT FOR PRECISION STAGING THROUGH EARLY AND ACCURATE DETECTION OF PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) LIVER METASTASIS. WE HAVE CREATED AN INNOVATIVE PLATFORM TECHNOLOGY USING A NEW CLASS OF PROTEIN-BASED MRI CONTRAST AGENTS (PROCAS) FOR CONTRAST-ENHANCED MRI. WE HAVE DEVELOPED AN EFFECTIVE APPROACH TO GENERATE PROTEIN CONTRAST AGENTS AGAINST AN ARRAY OF MOLECULAR BIOMARKERS INCLUDING COLLAGEN I (PROCA32.COLLAGEN). THIS EXTRACELLULAR MATRIX PROTEIN IS HIGHLY EXPRESSED IN THE TUMOR MICROENVIRONMENT AND ITS EXPRESSION LEVELS AND CROSSLINKING INCREASE UPON PROGRESSION. PROCA32.COLLAGEN EXHIBITS 10- TO 50-FOLD INCREASE IN BOTH R1 AND R2 RELAXIVITIES, COMPARED TO CLINICALLY-APPROVED GD3+ CONTRAST AGENTS, RESULTING IN EXCEPTIONAL IMAGING CAPABILITY THAT CAN DISCERN HETEROGENEOUS TISSUE SIGNALS VIA A DUAL MR IMAGING METHODOLOGY. PROCA32.COLLAGEN HAS STRONG COLLAGEN BINDING AFFINITY, ENABLES EARLY DETECTION OF SMALL LIVER METASTASES <0.2 MM AND ALLOWS FOR MAPPING TUMOR HETEROGENEITY IN SEVERAL MURINE XENOGRAFT TUMOR MODELS; A 100-FOLD IMPROVEMENT IN THE DETECTION LIMIT OVER CLINICAL CONTRAST AGENTS. PROCA32.COLLAGEN IS EXPECTED TO HAVE A LOW RISK RELATED TO METAL TOXICITY DUE TO ITS UNPRECEDENTED METAL BINDING SELECTIVITY AND KINETIC STABILITY, PROLONGED BLOOD RETENTION AND ADEQUATE BIODISTRIBUTION, WHICH PERMITS HIGH QUALITY IMAGING AT LOW DOSES. THIS PROPOSAL WILL TEST THE HYPOTHESIS THAT A NON- INVASIVE, IN VIVO MRI FOR ACCURATE STAGING OF PDAC THROUGH DETECTION OF SUBCLINICAL LIVER METASTASES CAN BE ACHIEVED BY FURTHER OPTIMIZING THE COLLAGEN TARGETED PROCA32.COLLAGEN+ CONTRAST AGENT. THIS SERIES OF PRECLINICAL STUDIES WILL PROVIDE DATA SUCH AS FORMULATION AND SAFETY PROFILE NEEDED TO SUBMIT AN FDA INVESTIGATIONAL NEW DRUG (IND) APPLICATION FOR AN EARLY PHASE CLINICAL TRIAL. THIS APPLICATION WILL IMPROVE DETECTION OF SUBCLINICAL METASTASIS AND HAVE BROAD IMPACT FOR PDAC.

DESIGN OF CALCIUM-SENSING RECEPTOR TARGETED THERA(G)NOSTIC AGENTS - SUMMARY CALCIUM-SENSING RECEPTOR (CASR), AS ONE OF THE FAMILY C G-PROTEIN-COUPLED RECEPTORS (GPCRS), LARGELY MODULATES EXTRACELLULAR CA2+ BY TRIGGERING MULTIPLE INTRACELLULAR SIGNALING PATHWAYS. CASR IS EXPRESSED IN THE KIDNEYS, PARATHYROID GLANDS, COLON, AND OTHER CELLS, REPRESENTING ITS DIFFERENTIAL YET KEY ROLES IN RESPONDING TO EXTRACELLULAR CALCIUM, AMINO ACIDS, AND METABOLITES, IN DIFFERENT ORGANS. NUMEROUS DISEASES HAVE BEEN ATTRIBUTED TO CALCIUM MISHANDLING (E.G.,NEPHROLITHIASIS) AND SECONDARY HYPERPARATHYROIDISM (HPT), WITH OVER 200 CASR MUTATIONS THAT CAN SIGNIFICANTLY ALTER CASR EXPRESSION. THERE IS A PRESSING UNMET MEDICAL NEED TO DEVELOP POSITIVE ALLOSTERIC MODULATORS FOR PKD AND OTHER DISEASES WITH CELLULAR AND ORGAN SPECIFICITY AND REDUCED LIVER TOXICITY. THE MAJOR BARRIERS TO DEVELOPING EFFECTIVE DRUGS ARE A LACK OF KNOWLEDGE OF TEMPO-SPATIAL QUANTITATIVE EXPRESSION OF CASR IN THE BODY AND ITS CHANGES DURING DISEASE PROGRESSION AND REGRESSION. WE RECENTLY DETERMINED THE FIRST CRYSTAL STRUCTURE OF THE HUMAN CASR EXTRACELLULAR DOMAIN (ECD) WITH A BOUND TRYPTOPHAN DERIVATIVE (TNCA) AT 2.1 Å, A POTENTIAL POSITIVE ALLOSTERIC MODULATOR. THIS LED US TO DESIGN AND SYNTHESIZE A DRUG CANDIDATE TNCA THAT HAS STRONG AGONIST ACTIVITY AND SPECIFICITY TO CASR. THE GOAL OF THIS PHASE I PROPOSAL IS TO ACQUIRE PROOF-OF-CONCEPT RESULTS FOR DESIGN AND GENERATION OF TNCA ANALOGS HAVING OPTIMAL DRUG ACTIVITY AND CELLULAR SPECIFICITY AS AGONISTS AND ANTAGONISTS. IN ADDITION, WE WILL ALSO DEVELOP CASR TARGETED PET AGENTS FOR RENAL IMAGING IN A PRECLINICAL PKHD1PCK RAT MODEL (STUDIES IN THIS MODEL LED TO CLINICAL TRIALS OF AN FDA-APPROVED PKD THERAPEUTIC). AIM 1 IS TO OPTIMIZE AND DETERMINE TNCA ANALOGS WITH BEST CASR DRUG ACTIVITY AS AGONISTS & ANTAGONISTS. AIM 2 IS TO PERFORM IN VIVO MAPPING OF CASR DISTRIBUTION IN PKD (PKHD1PCK) RAT MODEL BY MICROPET IMAGING. PHASE II WILL FURTHER VALIDATE DRUG EFFICACY OF TNCA ANALOGS, IMAGING CAPABILITY FOR CASR ASSOCIATED KIDNEY DISEASES USING ANIMAL MODELS, AND SAFETY PROFILE FOR AN EIND. SUCCESS IN OUR PROPOSED STUDIES WILL PROVIDE ESSENTIAL RESULTS FOR A NOVEL AVENUE IN THE DEVELOPMENT OF CASR SPECIFIC DRUG CANDIDATES FOR THE EFFECTIVE TREATMENT OF VARIOUS CASR ASSOCIATED HUMAN DISEASES USING NEWLY AVAILABLE STRUCTURES AND PATENTED METHODOLOGY. IN ADDITION, THE DEVELOPED PET IMAGING AGENTS WILL ENABLE US TO MAP CASR DISTRIBUTION AND HETEROGENEOUS EXPRESSION DURING PKD DISEASE PROGRESSION AND REGRESSION IN VIVO, AS A COMPANION DIAGNOSTIC FOR PATIENT RISK STRATIFICATION, AND FOR OUTCOME PREDICTION AND NOVEL DRUG DEVELOPMENT. WE ANTICIPATE THAT THE PROPOSED STUDIES WILL FOSTER THE DEVELOPMENT OF TRANSFORMATIVE NOVEL MARKERS OF THE DISEASE ACTIVITY THAT WILL ENHANCE CLINICAL CARE OF PKD AND OTHER KIDNEY DISEASES.

PRECISION MRI WITH A NOVEL PROTEIN CONTRAST AGENT FOR EARLY DETECTION AND STAGING OF LUNG FIBROSIS - SUMMARY LUNG DISEASES, SUCH AS INTERSTITIAL LUNG DISEASES INCLUDING IDIOPATHIC PULMONARY FIBROSIS (IPF), CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), AND ACUTE VIRAL INFECTION ARE MAJOR LEADING CAUSES OF DEATH WORLDWIDE. THE RECENT INCREASE DUE TO AIR POLLUTION, TOBACCO SMOKING IS AND USE OF E-CIGARETTES HAS GROWN RAPIDLY IN THE US CONTRIBUTING TO A SHARP INCREASE IN CHRONIC LUNG DISEASES. IPF IS CHARACTERIZED BY INCREASED EXTRACELLULAR MATRIX (ECM), INCLUDING COLLAGEN, WITHIN THE LUNG, CAUSING IRREVERSIBLE LUNG DAMAGE. WHILE HIGH-RESOLUTION COMPUTED TOMOGRAPHY (HRCT) IS COMMONLY USED TO DIAGNOSE IPF. THIS METHOD CAN PRODUCE LARGE VARIATION AND ERROR (UP TO 50%) AND IS UNABLE TO DETECT EARLY-STAGE LUNG FIBROSIS. AS THE PRESENCE OF SYMPTOMS DOES NOT ALWAYS INDICATE DISEASE TYPE, DIAGNOSIS IS USUALLY OBTAINED AT A MUCH LATER STAGE. DESPITE THE ADVANTAGES OF MRI, WHICH INCLUDES DEPOSITION DEPTH PENETRATION, LACK OF IONIZING RADIATION, AND HIGH RESOLUTION, THE APPLICATION OF MRI TO LUNG IMAGING HAS BEEN VERY CHALLENGING DUE TO REDUCED (10X LOWER) DENSITY OF THE LUNGS COMPARED TO OTHER TISSUES AND LIMITATIONS FROM EXTENSIVE DIPOLAR INTERACTIONS WITH MOLECULAR OXYGEN IN THE LUNGS. THEREFORE, THERE IS A PRESSING UNMET MEDICAL NEED TO DEVELOP NONINVASIVE IMAGING METHODOLOGIES AND CONTRAST AGENTS TO DETECT EARLY STAGES OF LUNG FIBROSIS, AND STAGE FIBROSIS SEVERITY BASED ON PROGRESSION OF IPF.

DESIGN OF CALCIUM-SENSING RECEPTOR TARGETED THERA(G)NOSTIC AGENTS - SUMMARY CALCIUM-SENSING RECEPTOR (CASR), AS ONE OF THE FAMILY C G-PROTEIN-COUPLED RECEPTORS (GPCRS), LARGELY MODULATES EXTRACELLULAR CA2+ BY TRIGGERING MULTIPLE INTRACELLULAR SIGNALING PATHWAYS. CASR IS EXPRESSED IN THE KIDNEYS, PARATHYROID GLANDS, COLON, AND OTHER CELLS, REPRESENTING ITS DIFFERENTIAL YET KEY ROLES IN RESPONDING TO EXTRACELLULAR CALCIUM, AMINO ACIDS, AND METABOLITES, IN DIFFERENT ORGANS. NUMEROUS DISEASES HAVE BEEN ATTRIBUTED TO CALCIUM MISHANDLING (E.G.,NEPHROLITHIASIS) AND SECONDARY HYPERPARATHYROIDISM (HPT), WITH OVER 200 CASR MUTATIONS THAT CAN SIGNIFICANTLY ALTER CASR EXPRESSION. THERE IS A PRESSING UNMET MEDICAL NEED TO DEVELOP POSITIVE ALLOSTERIC MODULATORS FOR PKD AND OTHER DISEASES WITH CELLULAR AND ORGAN SPECIFICITY AND REDUCED LIVER TOXICITY. THE MAJOR BARRIERS TO DEVELOPING EFFECTIVE DRUGS ARE A LACK OF KNOWLEDGE OF TEMPO-SPATIAL QUANTITATIVE EXPRESSION OF CASR IN THE BODY AND ITS CHANGES DURING DISEASE PROGRESSION AND REGRESSION. WE RECENTLY DETERMINED THE FIRST CRYSTAL STRUCTURE OF THE HUMAN CASR EXTRACELLULAR DOMAIN (ECD) WITH A BOUND TRYPTOPHAN DERIVATIVE (TNCA) AT 2.1 Å, A POTENTIAL POSITIVE ALLOSTERIC MODULATOR. THIS LED US TO DESIGN AND SYNTHESIZE A DRUG CANDIDATE TNCA THAT HAS STRONG AGONIST ACTIVITY AND SPECIFICITY TO CASR. THE GOAL OF THIS PHASE I PROPOSAL IS TO ACQUIRE PROOF-OF-CONCEPT RESULTS FOR DESIGN AND GENERATION OF TNCA ANALOGS HAVING OPTIMAL DRUG ACTIVITY AND CELLULAR SPECIFICITY AS AGONISTS AND ANTAGONISTS. IN ADDITION, WE WILL ALSO DEVELOP CASR TARGETED PET AGENTS FOR RENAL IMAGING IN A PRECLINICAL PKHD1PCK RAT MODEL (STUDIES IN THIS MODEL LED TO CLINICAL TRIALS OF AN FDA-APPROVED PKD THERAPEUTIC). AIM 1 IS TO OPTIMIZE AND DETERMINE TNCA ANALOGS WITH BEST CASR DRUG ACTIVITY AS AGONISTS & ANTAGONISTS. AIM 2 IS TO PERFORM IN VIVO MAPPING OF CASR DISTRIBUTION IN PKD (PKHD1PCK) RAT MODEL BY MICROPET IMAGING. PHASE II WILL FURTHER VALIDATE DRUG EFFICACY OF TNCA ANALOGS, IMAGING CAPABILITY FOR CASR ASSOCIATED KIDNEY DISEASES USING ANIMAL MODELS, AND SAFETY PROFILE FOR AN EIND. SUCCESS IN OUR PROPOSED STUDIES WILL PROVIDE ESSENTIAL RESULTS FOR A NOVEL AVENUE IN THE DEVELOPMENT OF CASR SPECIFIC DRUG CANDIDATES FOR THE EFFECTIVE TREATMENT OF VARIOUS CASR ASSOCIATED HUMAN DISEASES USING NEWLY AVAILABLE STRUCTURES AND PATENTED METHODOLOGY. IN ADDITION, THE DEVELOPED PET IMAGING AGENTS WILL ENABLE US TO MAP CASR DISTRIBUTION AND HETEROGENEOUS EXPRESSION DURING PKD DISEASE PROGRESSION AND REGRESSION IN VIVO, AS A COMPANION DIAGNOSTIC FOR PATIENT RISK STRATIFICATION, AND FOR OUTCOME PREDICTION AND NOVEL DRUG DEVELOPMENT. WE ANTICIPATE THAT THE PROPOSED STUDIES WILL FOSTER THE DEVELOPMENT OF TRANSFORMATIVE NOVEL MARKERS OF THE DISEASE ACTIVITY THAT WILL ENHANCE CLINICAL CARE OF PKD AND OTHER KIDNEY DISEASES.

A TREATMENT DRUG FOR TRIPLE NEGATIVE BREAST CANCER - SUMMARY TRIPLE NEGATIVE BREAST CANCERS (TNBC) ARE DEVASTATING DISEASES WITH A MEDIAN SURVIVAL OF LESS THAN 1-YEAR FOR PATIENTS WITH METASTATIC DISEASE. TNBC PATIENTS WITH TUMOR OF HIGH FIBROTIC STROMA HAVE EVEN WORSE PROGNOSIS. THERE ARE NO SPECIFIC TARGETED THERAPIES AVAILABLE FOR TNBC, AND THE ONLY TREATMENT OPTION IS BROADLY CYTOTOXIC CHEMOTHERAPY DRUGS, DESPITE LESS EFFECTIVE AND STRONG UNWANTED SIDE EFFECTS OF SUCH DRUGS. ONE MAJOR BARRIER TO EFFICACY OF ANTI-TUMOR THERAPEUTICS IS THE DENSE FIBROTIC STROMAL AND DYSREGULATED TUMOR BLOOD VESSELS WHICH CONTRIBUTE TO FAILURE OF THERAPIES. EVIDENCE SUGGESTS THAT CANCER ASSOCIATED FIBROBLASTS (CAF) PRODUCE THE STROMAL COLLAGEN. THE ECM LAID DOWN BY CAF IS CONSIDERED TO BE ONE OF THE MAJOR CONTRIBUTORS OF RESISTANCE TO ESTABLISHED THERAPIES OF THE DISEASES. TNBC HAS HIGH ANGIOGENIC ACTIVITY. DENSE TUMOR VASCULATURE ASSOCIATED WITH A SHORTER TIME FROM DIAGNOSIS TO RELAPSE AND FROM RELAPSE TO DEATH. THE DYSREGULATED VESSEL STRUCTURE IN TNBC TUMOR OFTEN LEADS TO RESISTANCE TO BLOOD FLOW INTO TUMOR, WHICH IS ANOTHER IMPORTANT BARRIER FOR DRUG DELIVERY. DEPLETING CAF AND ABROGATING TUMOR ANGIOGENESIS COULD SIGNIFICANTLY IMPROVE EFFICACY OF EXISTING TNBC CANCER TREATMENTS. HOWEVER, CURRENTLY, THERE ARE NO APPROVED THERAPIES THAT ARE ABLE TO DEPLETE CAF AND TUMOR ANGIOGENESIS IN TNBC CANCER. WE HAVE DEVELOPED A NOVEL THERAPEUTIC PROTEIN (PROAGIO) USING RATIONAL PROTEIN DESIGN. PROAGIO IS DESIGNED TO TARGET INTEGRIN V3 AT A NOVEL SITE (NOT THE LIGAND BINDING SITE). PROAGIO SPECIFICALLY INDUCES APOPTOSIS OF INTEGRIN V3 EXPRESSING CELLS WITH HIGH EFFICACY BY A NOVEL MECHANISM OF DRUG ACTION (RECRUITING & ACTIVATING CASPASE 8 AT CYTOPLASMIC DOMAIN OF). WE REASONED THAT, SINCE BOTH CAF AND ANGIOGENIC ENDOTHELIAL CELLS (AEC) EXPRESS HIGH LEVELS OF INTEGRIN V3, AND SINCE PROAGIO IS VERY EFFECTIVE IN INDUCING APOPTOSIS OF INTEGRIN V3 EXPRESSING CELLS, PROAGIO SHOULD BOTH DEPLETE CAF, ELIMINATE INTRATUMORAL ANGIOGENIC BLOOD VESSELS IN AND AROUND TNBC TUMORS. THIS UNIQUE STRATEGY MAY PROVE ADVANTAGEOUS IN TREATMENT OF TNBC. THE MAIN OBJECTIVE OF THIS DIRECT PHASE II SBIR APPLICATION IS TO GENERATE A DEFINITIVE DATASET TO ENABLE THE DEVELOPMENT OF PROAGIO AS A VIABLE THERAPEUTIC OPTION FOR TNBC PATIENTS. CHARACTERIZATION OF THE TOXICITY AND TOLERABILITY AND DETERMINE THE MAXIMUM TOLERATED DOSE (MTD) AND RECOMMENDED PHASE II DOSE (RP2D) OF PROAGIO AS A SINGLE AGENT IS ON-GOING. AIM 1 WILL CHARACTERIZE THE TOXICITY AND TOLERABILITY AND DETERMINE THE RECOMMENDED PHASE II DOSE (RP2D) OF PROAGIO IN COMBINATION WITH GEMCITABINE (GEM). AIM 2 WILL OBTAIN PRELIMINARY ANTI-CANCER ACTIVITY DATA OF PROAGIO AND PROAGIO + GEM IN TNBC PATIENTS. AIM 3 WILL ANALYZE THE EFFECTS OF PROAGIO IN PATIENT TUMOR TO VALIDATE THE MECHANISM OF DRUG ACTION IN PATIENTS. THIS CLINICAL STUDY PROJECT WILL EXPLORE NEW THERAPEUTIC AVENUE FOR TNBC PATIENTS. OUR GOAL IS THAT, THROUGH OUR STUDY, WE WILL INTRODUCE A NEW TREATMENT APPROACH FOR TNBC WITH NOVEL MECHANISM OF DRUG ACTION.

DETECTION AND STAGING OF LIVER FIBROSIS BY PRECISE MRI (PMRI) - ABSTRACT CHRONIC LIVER INJURY DUE TO ALCOHOL, METABOLIC DYSFUNCTION, VIRAL HEPATITIS, OR AUTOIMMUNE DISEASE RESULTS IN LIVER INFLAMMATION AND FIBROSIS. LIVER FIBROSIS WILL PROGRESS TO CIRRHOSIS WHICH IS ESTIMATED TO AFFECT 1–2% OF THE WORLD’S POPULATION. THE MAJOR CLINICAL CONSEQUENCES OF CIRRHOSIS ARE LIVER FAILURE AND HEPATOCELLULAR CARCINOMA (HCC), BOTH OF WHICH INCREASE THE RISK OF DEATH. ALCOHOLIC LIVER DISEASE (ALD) IS A LEADING CAUSE OF LIVER DISEASE AND LIVER-RELATED DEATHS GLOBALLY, PARTICULARLY IN DEVELOPED NATIONS. THE CURRENT GOLD STANDARD DIAGNOSTIC METHOD, LIVER BIOPSY, HAS MANY LIMITATIONS INCLUDING SAMPLING ERROR AND HIGH INTER-OBSERVER VARIABILITY WITH A 33% ERROR RATE EVEN FOR THE DIAGNOSIS OF ADVANCED STAGES OF LIVER FIBROSIS. IN ADDITION, LIVER BIOPSY IS AN INVASIVE, PAINFUL AND EXPENSIVE PROCEDURE WITH THE RISK OF COMPLICATIONS INVOLVING HOSPITALIZATION AND MORTALITY. LIVER BIOPSY IS THEREFORE OF LIMITED USE FOR SCREENING OR MONITORING DISEASE PROGRESSION. NONE OF SEVERAL TECHNOLOGIES INVESTIGATED OFFERS THE DESIRED SENSITIVITY AND SPECIFICITY FOR THE DETECTION OR STAGING OF FIBROSIS. THE NOVEL CONTRAST AGENT DEVELOPED IN OUR LABORATORY, PROCA32.COLLAGEN1, COMBINED WITH INNOVATIVE PROCESSING TECHNIQUES MADE POSSIBLE BY THIS AGENT, PROMISE TO SIGNIFICANTLY ENHANCE THE PRECISION OF MRI IN DIAGNOSING AND MONITORING LIVER FIBROSIS. WE HAVE SUCCESSFULLY MET ALL OF THE PROPOSED AIMS AND MILESTONES FOR PHASE I TO OBTAIN PROOF-OF-PRINCIPLE EVIDENCE TO ACHIEVE EARLY DETECTION OF LIVER FIBROSIS WITH SIGNIFICANTLY IMPROVED SENSITIVITY AND SELECTIVITY. WE HAVE DEMONSTRATED THAT PROCA32.COLLAGEN DETECTS IN VIVO LIVER FIBROSIS IN MOUSE MODELS. IN THIS PHASE II PROJECT, WE WILL OPTIMIZE AFFINITY FOR COLLAGEN BINDING AND BIODISTRIBUTION AND VALIDATE IN VIVO CAPABILITY IN THE EARLY DETECTION AND STAGING OF LIVER FIBROSIS. WE WILL OBTAIN ESSENTIAL DATA REQUIRED FOR FILING IND APPLICATIONS LEADING TO CLINICAL TRANSLATION.

DEVELOPMENT OF A NOVEL MRI CONTRAST AGENT FOR EARLY DETECTION OF ALCOHOLIC STEATOHEPATITIS