Translating a Novel Assay for Risk Stratification of Potentially Malignant Pancreatic Cystic Lesions

Abstract public domain (NIH RePORTER)

1. Executive Summary Pancreatic cancer has a depressing 5-year survival rate estimated at 12%. The dismal outlook is due to myriad issues, including the fact that patients are asymptomatic in early stages combined with the lack of screening tools. One window to earlier detection is through pancreatic cystic lesions detected incidentally in routine imaging analyses for diagnosis of unrelated disorders. These patients are at an increased risk for pancreatic cancer. Unfortunately, imaging is insufficient for distinguishing benign cysts from those likely to progress to cancer forcing clinicians to rely on biochemical analyses of acellular cyst fluid. The current assays display poor accuracy and require large volumes of fluid not always available limiting their clinical utility. There is a profound unmet clinical need for diagnostics that accurately grade dysplasia in mucinous pancreatic cyst fluids. The objective of current SBIR Phase I grant (CA277913-01A1 effective May 2023) is to develop and conduct clinical translation of a diagnostic assay panel using ultrasensitive detection methods to grade dysplasia in pancreatic cysts. This assay will stratify patients most at risk for progression to cancer thus providing high clinical utility. The platform: Amplified Sciences has utilized a proprietary platform to translate to the company roscopy platform is robust and readily adaptable to other proteolytic enzymes as needed for this grant. To date, the company has made strong progress on the Aims. The team has built a new direct-detection SERS ligand binding platform (prototype stage) for two of the biomarkers, and adapted the existing enzymatic activity assay for a third. Over 90 retrospective clinical samples have been procured, and all clinical testing and statistical analysis should be complete by end of Phase 1. The company is on track -ready assay protocol for full clinical validation and algorithm development in a Phase 2 grant. Several challenges have been met and overcome. Technical challenges including delays in the supply chain have been overcome by securing new suppliers for chemicals, etc. Administrative challenges include delays from securing invoices from vendors, so more frequent reminders have been implemented. Finally, commercial challenges include limited funding for market research with doctors and payers to gain the insights needed to prepare for commercial launch and challenges in accessing clinicians at large academic centers. To overcome this, the company has leveraged current clinical investigators for introductions. Finally, the current capital market for fundraising is very challenging so the company has supplemented investor funds with non-dilutive funding totaling $1M of research use only revenue and state and federal grants. Specific Aims: Aim 1: Validate a combination of three (3) protease activity markers to accurately distinguish low- grade from high-grade mucinous pancreatic cystic neoplasms. Aim 2: To develop quantitative assays for a panel of three (3) markers in low-volume mock cyst fluid samples that enable diagnostic power evaluation in a clinical sample cohort. Below employees are committed to I-Corps requirements: Diana Caldwell, CEO and Co-founder (C- er) Daniel Sheik, PhD, Director of Research and Technology John R. Ridge, Commercial and Market Access Director (Industry Expert)

Public health relevance public domain (NIH RePORTER)

2. I-Corps Team The I-Corps team will be formed with three key members of the leadership team. The team has actively been involved in product development and is currently building commercialization strategies for the launch of this assay via a targeted “early access” launch in 2025. All team members have previous experience with startups in the life science industry. Members also have commercial launch, market research, and diagnostics industry experience which will allow the team to effectively carry out the requirements of the I-Corps grant. The Corporate Officer representative will be Diana Caldwell, CEO, who is an accomplished and recognized serial life science entrepreneur with over 25 years of experience in both large corporate and small entrepreneurial environments. Throughout her career, she has successfully driven organizational growth, developed, and implemented strategic business plans, raised capital, launched novel life science products, and directed successful teams. Diana has been leading Amplified Sciences for three years, has decades of life science product launch experience, and has in-depth knowledge of the technologies and the markets that we will be addressing in carrying out the objections of the I-Corps grant. Dan Sheik, PhD, Director of Research and Technology at Amplified Sciences, is the PI on this working to improve the diagnostic sensitivity of assays using proprietary rhodamine dyes and the ultrasensitive Surface- on early detection of pancreatic cancer to improve the quantitative resolution of biomarkers to give a clearer picture of pancreatic cyst malignancy. Dan’s assay development experience and scientific skills provide the project with the technical expertise that will be necessary to meet the objectives of the I-Corps grant. John R. Ridge will round out the team as the Industry Expert. John joined Amplified Sciences as 20 years of experience working in Market Access and cross functionally with commercial functions for Medical Device and Diagnostics companies. Additionally, he worked on the payer side early in his career thus bringing a well rounded industry perspective to the challenges and opportunities in successful commercial launch and reimbursement efforts. Before joining Amplified Sciences as an employee, he served as an Advisor for the company and is familiar with the company’s new technologies and will use that knowledge and prior diagnostic product launch experience to contribute effectively to the team’s efforts. The I-Corps grant will be valuable to the team as it will allow us to further build on initial research that was completed by student interns. That project identified potential key targets that we could reach out to with the discovery questions, and that would allow us to gain key data to assess customer needs, understand commercial adoption barriers, explore the value this innovation provides, and develop potential options to position the new assay. All team members will be able to meet the time intensive time requirements outlined in the program. This proposed team is also able to benefit maximally from the I-Corps program as it has already conducted background research into competitive IP landscape, competitors’ publications, customer needs, value proposition, market assessment and reimbursement models. With new knowledge and information gained from the I-Corps program, the team is capable of and looks forward to a thorough vetting from potential customers and industry experts of these early concepts and refining, realigning and/or pivoting their initial plan to maximize commercialization success. 3. Potential Commercial Impact and Target Customers Research indicates that the pancreatic cancer diagnostics US market is a robust +$2.9 billion (TAM). Over 3 million patients are diagnosed annually in the US with a pancreatic cyst, and it is -FNA procedure to sample cyst fluid. A conservative

Public health relevance public domain (NIH RePORTER)

estimate of ~300,000 patients tested annually yields a serviceable available market (SAM) of at least $300 million in the US. Amplified Sciences’ sales revenue for the rule-in high-grade vs. low- grade assay forecast at $80 million five years post launch. This represents 20,000 tests and a 20- 25% market share. The proposed customers would be centers of specialty gastroenterologists (30-50), advanced/interventional endoscopists, specialty fellows, pancreatic cyst clinics, and high-risk pancreatic cancer centers. Within this market, there is a large unmet medical need for earlier detection of pancreatic cancer given that it is the third deadliest cancer with the highest mortality rate in the US. Patients diagnosed with pancreatic cystic lesions are at increased risk for pancreatic cancer and represent a window to early detection, and monitoring and surgery on these patients is expensive and associated with complications and morbidity. One of our specific plans for the I-Corps program is to conduct interviews with the customer targets and influencers. Clinician influencers include pancreatic biliary surgeons, oncologists, and hospital pathologists. Although the customer physician targets are clear, there are also key decision makers with commercial and government payers. Their role is to determine if a new novel assay should be covered and paid when a bill is submitted for a lab developed test by the diagnostic company. In the commercialization of a new assay, it is critically important to assess payers’ thoughts and gain their inputs on the clinical value of the assay. As such, they will need to be a target of the project as well. In addition, there is no current standard testing for grading dysplasia, with multiple attempts showing little promise. In fact, peer reviewed studies have demonstrated that standard cytology fails over 70% of the time in this disease state due to acellularity of the cyst fluid sample and the low volume of fluid that is successfully procured during an EUS-FNA. Given the lack of laboratory- based solutions, clinicians and payers are very receptive to novel diagnostics with increased sensitivity and specificity vs. currently available diagnostic competitors that can provide improved outcomes and cost savings. A competitive advantage rests in our ability to provide clinicians access to an assay that delivers higher content diagnostic information about the malignant potential of the mucinous pancreatic cyst population. By analyzing competitors and similar lab developed test models across other cancers, a target price range between $2000 - $3000 has been established but needs to be confirmed. This price point has been established considering several factors: material and production cost, reimbursement of similar assays, and likelihood to be able to cross-walk to an existing test in a different disease state. It is important to note that the price point is sensitive to the assay reimbursement rate that would be determined by the Centers for Medicare and Medicaid Services or the Medicare Administrative Contractors. We are open to revisiting these additional factors and possibly revising this price point based on interviews with a much larger pool of stakeholders of the assay. 4. Project Plan - readiness, and adaptability to standard clinical lab test processes. Amplified Sciences seeks to -certified lab that are novel and demonstrate superior accuracy with a keen desire to stretch the capabilities and acceptance of SERS as a detection platform. These efforts to validate biomarkers of dysplasia in mucinous pancreatic cysts in this Phase 1 are expected to produce assays that have demonstrated clinical proof of concept in a small retrospective cohort. These assays will be ready for further analytical validation and larger hypothesis, Amplified Sciences will use the protocols developed in AIM 2 to reanalyze the 100 clinical samples from AIM 1 and compare the diagnostic results after optimization with a second

Public health relevance public domain (NIH RePORTER)

nested-cross validation study to those of AIM 1. It is expected that the results using Amplified Sciences’ assay platform will prove to be at least non-inferior to the results found in the AIM 1 validation studies. Importantly, the new assays will use low sample volumes, allowing for the multi-analyte panel to be tested on -volume analyses prevent interruption of current standards of care while this panel is introduced to the clinic. Currently we are continuing optimization of marker assays to our SERS platform before final clinical sample testing. This is expected to be completed prior to submission of our Phase II application. Assuming that these results are promising, the company is prepared to execute the -certified lab in Irvine, California. The company will execute the analytical validation and clinical validation protocols required to regulatory milestone, the company will prepare for a targeted early-access launch to advanced endoscopist gastroenterologists targeted by the end of 2025. Phase II will combine a set of multiplexed low-volume assays ready to commercialize as lab -site prospective sample (n = 300) collection study.