Clinical Research Reporting AEs Protocol Review

1: From a CRC perspective, review the protocol (found in the Protocol module folder) and answer the following: • What is the withdrawal criteria and what steps would need to be taken to ensure the participant was safe after withdrawal? How would you document this? • Look at the Schedule of events (8.1 and 8.2), it lists that AEs should be assessed often. How would you plan this type of assessment, how would you document? • Are there adequate safety parameters (labs, tests, examinations) to ensure the participant is safe during the study? What are the safety parameters? 2: From a CRA perspective, review the protocol and answer the following: • What types of data would you look for when assessing Adverse Events (both AEs and SAEs) during your routine monitoring visit? • Look at the information found in 9.2, where would you look to find the data for each bullet point? • Look at 9.6, how would information from the safety committee be documented? CLINICAL TRIAL PROTOCOL NCR-001-1 Version and date: Version 4, 18 February 2018 Sponsor: Sponsored Therapeutics 1635 East 18th Street City, ST 12345 us e on Protocol Number: ly Formal Title: NCR-001: A Dose Comparison of NCR-001 to Defy in Healthy Volunteers Specialist Doctor, MD Pennant University Medical Center 1234 Address Avenue City, ST 12345 rC University of Medical Center 12345 Medical Center City, ST 67890 Fo Imaging Core Lab: R C 27 0 Study Principal Investigator: 1 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL Investigator Protocol Agreement PROTOCOL NO: NCR-001-1 PROTOCOL TITLE: NCR-001: A Dose Comparison of NCR-001 to Defy in Healthy Volunteers By signing, I confirm that I/my staff have read and understand this protocol and agree to comply with the conduct and terms of this study. I/we have agreed to abide by the following responsibilities: ly C  27 0     on  e   To conduct the study in compliance with this protocol, any future amendments, any conditions of the governing reviewing EC/IRB or regulatory agency. To supervise all testing involving human subjects. To abide by Good Clinical Practices (GCP), and all applicable regional or national regulations. Ensure that the requirements for obtaining informed consent from each study participant or their legal representative are met. To report any Serious Adverse Events in a timely manner as required by the protocol. To report any Serious Adverse Events to the IRB in a timely manner. To report Non-serious Adverse Events as required by the protocol. Clinical Research Reporting AEs Protocol Review

To maintain confidentiality and assure security of confidential documents that include but are not limited to: the protocol, case report forms, Investigator’s Brochure, final study documents, manuscript drafts, unpublished data, correspondence, etc. To cooperate fully with any regulatory agency audit. us  rC R Principal Investigator (name): _________________________________________ Fo Principal Investigator (signature): ______________________________________ 2 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL SUSAR on Fo rC R C 27 USCA e IRB IV LVO MB MI NYHA OFP SAE us HEENT Adverse Event American Society of Echocardiography Chronic Obstructive Pulmonary Disease Electronic Case Report Form Electrocardiogram Food and Drug Administration Good Clinical Practices Head, eyes, ears, nose, and throat examination Institutional Review Board Intravenous Left Ventricular Opacification Minispheres Mechanical Index New York Heart Association Octafluoropropane Serious Adverse Event Suspected Unexpected Serious Adverse Reactions Ultrasound Contrast Agents 0 AE ASE COPD eCRF EKG FDA GCP ly TABLE OF ABBREVIATIONS 3 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL Table of Contents CLINICAL TRIAL PROTOCOL ………………………………………………………………………………….. 1 Investigator Protocol Agreement……………………………………………………………………………………. 2 TABLE OF ABBREVIATIONS …………………………………………………………………………………….. 3 1. BACKGROUND ……………………………………………………………………………………………………….. 7 e on ly 1.1 Contrast Enhanced Echocardiography ……………………………………………………………………….. 7 1.2 Rationale for use in Endocardial Border Delineation …………………………………………………… 7 1.3 NCR-001: Acoustically active minispheres ………………………………………………………………….. 7 1.3.1 Preclinical efficacy studies in echocardiography ……………………………………………………………… 7 1.3.2 Non-clinical pharmacology and toxicology …………………………………………………………………….. 8 1.4 Pharmacokinetics and Pharmacodynamics of NCR-001 in Human Subjects ……………………….. 8 1.4.1 Mechanism of Action …………………………………………………………………………………………………… 8 1.4.2 Pharmacokinetics ………………………………………………………………………………………………………… 9 1.4.3 Distribution…………………………………………………………………………………………………………………. 9 1.4.4 Metabolism …………………………………………………………………………………………………………………. 9 1.4.5 Elimination…………………………………………………………………………………………………………………. 9 1.4.6 Special Populations ……………………………………………………………………………………………………… 9 us 2. OBJECTIVES …………………………………………………………………………………………………………… 9 2.1 Primary Objectives…………………………………………………………………………………………………………. 10 2.2 Secondary Objectives ……………………………………………………………………………………………………… 10 0 3. SUBJECT SELECTION ………………………………………………………………………………………….. 10 C 27 3.1 Inclusion Criteria……………………………………………………………………………………………………………. 10 3.2 Exclusion Criteria…………………………………………………………………………………………………………… 10 3.3 Withdrawal Criteria……………………………………………………………………………………………………….. 11 3.4 Replacement of Subjects …………………………………………………………………………………………………. 11 R 4. REGISTRATION PROCEDURES…………………………………………………………………………… 12 rC 4.1 Registration Process ……………………………………………………………………………………………………….. 12 5. TREATMENT PLAN………………………………………………………………………………………………. 12 Fo 5.1 Study Treatments …………………………………………………………………………………………………………… 12 5.1.1 Bolus ……………………………………………………………………………………………………………………….. 12 5.1.2 IV Infusion ……………………………………………………………………………………………………………….. 12 5.2 Safety Guidelines for Study Treatments…………………………………………………………………………… 13 5.2.1 Bolus ……………………………………………………………………………………………………………………….. 13 5.2.2 IV infusion………………………………………………………………………………………………………………… 13 5.3 Duration of Therapy ……………………………………………………………………………………………………….. 13 5.3.1 Bolus ……………………………………………………………………………………………………………………….. 13 5.3.2 IV Infusion ……………………………………………………………………………………………………………….. 13 5.4 Duration of Follow-up …………………………………………………………………………………………………….. 13 6. DOSING DELAYS/DOSE MODIFICATIONS …………………………………………………………. 13 7. STUDY ASSESSMENTS …………………………………………………………………………………………. 14 7.1 Screening and Informed Consent …………………………………………………………………………………….. 14 7.2 Specific Procedures…………………………………………………………………………………………………………. 14 7.2.1 Height and Weight …………………………………………………………………………………………………….. 14 7.2.2 Vital Signs ………………………………………………………………………………………………………………… 14 7.2.3 Medical History …………………………………………………………………………………………………………. 14 4 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL 7.2.4 Concomitant Mediations …………………………………………………………………………………………….. 14 7.2.5 Clinical Laboratory Assessments …………………………………………………………………………………. 14 7.2.6 Physical Examination …………………………………………………………………………………………………. 15 7.2.7 12-lead Electrocardiogram (EKG) ……………………………………………………………………………….. 15 7.2.8 Echocardiograms ……………………………………………………………………………………………………….. 16 7.3 Assessments Performed by Study Visit…………………………………………………………………………….. 16 7.3.1 Screening ………………………………………………………………………………………………………………….. 16 7.3.2 Dosing ……………………………………………………………………………………………………………………… 17 7.3.3 Discharge………………………………………………………………………………………………………………….. 17 7.4 Long Term Follow-up …………………………………………………………………………………………………….. 17 8. STUDY SCHEDULE OF EVENTS ………………………………………………………………………….. 18 9. DOSE LIMITING TOXICITIES, ADVERSE EVENTS, AND REPORTING REQUIREMENTS………………………………………………………………………………………………………. 22 us e on ly 9.1 Dose Limiting Toxicity (DLT) …………………………………………………………………………………………. 22 9.2 Definition of Adverse Events …………………………………………………………………………………………… 23 9.3 Reporting Procedures for Adverse Events ……………………………………………………………………….. 23 9.3.1 Follow-up of an AE ……………………………………………………………………………………………………. 23 9.4 Serious Adverse Events …………………………………………………………………………………………………… 24 9.4.1 Reporting procedures for SAEs……………………………………………………………………………………. 24 9.5 Routine Adverse Event Reporting …………………………………………………………………………………… 24 9.6 Stopping Rules ……………………………………………………………………………………………………………….. 24 9.7 Integrative Safety Analysis ……………………………………………………………………………………………… 25 0 10. PHARMACEUTICAL INFORMATION ………………………………………………………………… 25 R C 27 10.1 NCR-001 ………………………………………………………………………………………………………………………. 25 10.1.1 Storage and Preparation of the Investigational Product ………………………………………………. 25 10.1.2 Administration …………………………………………………………………………………………………………… 25 10.1.3 Dose Modifications ………………………………………………………………………………………………….. 25 10.1.4 Return and Destruction of the Investigational Product ………………………………………………… 25 11. MEASUREMENT OF EFFECT …………………………………………………………………………….. 26 rC 11.1 Echocardiographic Measurements…………………………………………………………………………………. 26 12. STATISTICAL CONSIDERATIONS …………………………………………………………………….. 27 Fo 12.1 Study Design/Endpoints ………………………………………………………………………………………………… 27 12.1.1 Primary endpoints ……………………………………………………………………………………………………. 27 12.2 Sample Size…………………………………………………………………………………………………………………… 28 12.3 Randomization Scheme …………………………………………………………………………………………………. 28 13. DATA REPORTING ……………………………………………………………………………………………… 28 13.1 Data Collection……………………………………………………………………………………………………………… 28 13.1.1 Data Collection at Sites …………………………………………………………………………………………….. 28 13.1.2 Echocardiogram collection ………………………………………………………………………………………… 28 13.1.3 Non-blinded image preparation for each echo study ……………………………………………………… 28 13.1.4 Blinded Review of Echocardiograms ………………………………………………………………………….. 29 13.2 Study Monitoring and Data Queries ………………………………………………………………………………. 29 13.3 Publication Policy …………………………………………………………………………………………………………. 29 13.4 Registration with a Clinical Trial Registry …………………………………………………………………….. 29 14. ETHICAL AND REULATORY REQUIRMENTS………………………………………………….. 29 14.1 Informed Consent …………………………………………………………………………………………………………. 29 14.2 Institutional Review Board ……………………………………………………………………………………………. 30 5 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL 14.3 Subject Confidentiality………………………………………………………………………………………………….. 30 14.4 Protocol Amendments …………………………………………………………………………………………………… 30 14.5 Study Termination………………………………………………………………………………………………………… 30 Fo rC R C 27 0 us e on ly 15. REFERENCES ………………………………………………………………………………………………………. 31 6 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL 1. BACKGROUND ® DEFY (perflupan lipid microspheres, Lateral Medical Imaging) is an ultrasound contrast indicated for use in subjects with suboptimal echocardiograms to opacify the left ventricular chamber and to improve the delineation of the left ventricular endocardial border. NCR-001 is ® similar to DEFY , with the exception of the removal of one lipid in the lipid blend and substitution of another lipid. This study is intended to evaluate the safety and efficacy of NCR001 using a single ascending dose design in healthy volunteers. 1.1 Contrast Enhanced Echocardiography Ultrasound is one of the most common imaging examinations and has advantages of absence of ionizing radiation, portability and relatively low cost (1). Ultrasound contrast agents are used ® on ly to improve the accuracy of ultrasound and DEFY (perflupan) is the world’s leading ultrasound contrast agent but perflupan has to be refrigerated and has a side effect of back pain (2). The investigators have developed a new, improved perflupan with potential for room temperature storage. us e 1.2 Rationale for use in Endocardial Border Delineation 27 0 Ultrasound contrast agents (USCA) are used to increase the backscatter (signal intensity of blood vessels and tissues to improve diagnostic accuracy) of ultrasound imaging (3). In the US, ultrasound contrast agents are FDA approved for echocardiography to improve endomyocardial border definition. C 1.3 NCR-001: Acoustically active minispheres Fo rC R NCR-001 reflects the ultrasound to provide strong back-scattering. NCR-001minispheres resonate with ultrasound to provide strong harmonic signals. NCR-001 minispheres will cavitate with higher levels of ultrasound and cavitation may be stable or inertial depending upon acoustic parameters and other factors. 1.3.1 Preclinical efficacy studies in echocardiography Imaging of NCR-001:  In vitro acoustic performance comparable/superior to DEFY  Echocardiography performed in 5 pigs ® ®  DEFY and NCR-001 provide comparable contrast enhancement at low and high MI.  No change in hemodynamics with NCR-001 Porcine cardiac imaging with NCR-001 shown in Figure 1.3.1 below: 7 NCR-001-1 Clinical Trial Protocol V4 Dated 18 February 2018 CONFIDENTIAL ly on e us 0 27 R C Figure 1.3.1 Porcine cardiac imaging with NCR-001. rC 1.3.2 Non-clinical pharmacology and toxicology Fo 1.3.2.1 Carcinogenesis, Mutagenesis, and Impairment of Fertility ® Studies with activated DEFY have not been performed to evaluate carcinogenic potential. ® Evidence of genotoxicity was not found in the following studies with activated DEFY : 1) bacterial mutagenesis assay (Ames assay), 2) in v . Clinical Research Reporting AEs Protocol Review