Phase 1/2A Study of Rintatolimod and IFN Alpha Regimen in Cancer Patients With COVID-19
Phase: PHASE1
Conditions: Hematopoietic and Lymphoid Cell Neoplasm, Malignant Solid Neoplasm, Symptomatic COVID-19 Infection Laboratory-Confirmed
Sponsor: Roswell Park Cancer Institute

Document Coversheet 
 
  
 
 Study Title:     Phase 1/2A Study of Rintatolimod and IFN alpha Regimen in Cancer Patients with 
Mild or Moderate COVID-19 Infection 
 
 
Institution/Site:  
Roswell Park Comprehensive Cancer Center 
Document (Approval/Update) Date: 
03-06-2025 
NCT Number: 
 NCT04379518 
IRB Number 
I 659920 
 
 
 
 
 

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Date: 02/13/2025; 3/6/2025 (IRB)
PROTOCOL TITLE:
Phase 1/2A Study of Rintatolimod and IFN alpha Regimen in Cancer Patients with 
COVID-19 
PROTOCOL NUMBER:
I-659920
PRINCIPAL INVESTIGATOR:
Igor Puzanov, MD
Judith and Stanford Lipsey Endowed Chair in Clinical Studies
Professor of Oncology and Medicine
Director, Center for Early Phase Clinical Trials
Co-Leader, Developmental Therapeutics Program
Chief, Early Phase Clinical Trials Division
Chief, Melanoma Service
Roswell Park Comprehensive Cancer Center
Elm & Carlton Streets, Buffalo, NY 14263
Tel.: +1.716.845.7505; E-mail: Igor.Puzanov@RoswellPark.org
VERSION NUMBER: 
12
DATE: 
02/13/2025
Confidentiality Statement
Any and all information presented in this document shall be treated as confidential and shall 
remain the exclusive property of the party (ies) mentioned above (Roswell Park Cancer Institute 
Corporation d/b/a Roswell Park Comprehensive Cancer Center). The use of such confidential 
information must be restricted to the recipient for the agreed purpose and must not be disclosed, 
published, or otherwise communicated to any unauthorized persons, for any reason, in any form 
whatsoever without the prior written consent of the party(ies) above.
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µg
microgram
µL
microliter
5-FU
5-fluorouracil 
ACE
angiotensin-converting-enzyme 
AE
adverse events
α
alpha
αDC
alpha Dendritic Cell
ALT
Alanine transaminase 
ANA
antinuclear antibodies 
ANC
absolute neutrophil count
AST
Aspartate transaminase 
ARDS
β
Acute respiratory distress syndrome
beta
BUN
blood urea nitrogen 
CBER
Center for Biologics Evaluation and Research
cc
cubic centimeter
CD3
cluster of differentiation 3
°C
degrees Celsius
CKM
chemokine modulatory
Cl
chloride
cm
centimeter
CO2
carbon dioxide
COX2
Cyclooxygenase-2
CRC
colorectal cancer
CRCLM
CRC liver metastasis
CRCM
metastatic colorectal cancer
CRS
Clinical Risk Score
CT
computed tomography 
CTCAE
Common Terminology Criteria for Adverse Events 
dL
deciliter
DLT
Dose Limiting Toxicity
DSMC
Data Safety Monitoring Committee
(ds)RNA
Double-stranded RNA
ECOG
Eastern Cooperative Oncology Group
EKG
Electrocardiography
°F
degrees Fahrenheit
FDA
F
g
gram

gamma
GI
gastrointestinal
GMP
good Manufacturing Practices
HCG
Human Chorionic Gonadotropin
HDI
High Dose Interferon
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HPRT
hypoxanthine phosphoribosyl transferase
ICH
immunohistochemistry
IFN
interferon
IL-2
interleukin-2
im
intramuscular
IND
Investigational New Drug
IRB
Institutional Review Board
ISH
in situ hybridization
IU
International Units
IV
intravenous
kg
kilogram
LPS
lipopolysaccharide
m2
meters squared
mg
milligram
min
minute
mL
milliliter
MRI
Magnetic Resonance 
mRNA
Messenger RNA 
NCI
National Cancer Institute
ng
nanogram
NSAID
non-steroidal anti-inflammatory drug
PET
Positron emission tomography 
PGE2
prostaglandin E2
po
RPCI
by oral route of administration
Roswell Park Cancer Institute
RP2D
recommended Phase 2 dose
SAE
severe adverse events
SGOT
Serum Glutamic Oxaloacetic Transaminase 
SGPT
Serum Glutamic Pyruvate Transaminase
SOC
standard of care
ssRNA
single-stranded RNA 
Teff
effector T cells
TLR
toll-like receptor
Treg
regulatory T cells
U
unit
ULN
upper limit of normal
UPCI
University of Pittsburgh Cancer Institute
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Table of Contents
1
Study Objectives......................................................................................................................5
2
Background..............................................................................................................................5
3
Inclusion and Exclusion Criteria ...........................................................................................11
4
Local and Study-Wide Number of Subjects ..........................................................................14
5
Local and Study-Wide Recruitment Methods .......................................................................15
6
Multi-Site Research ...............................................................................................................15
7
Study Timelines.....................................................................................................................15
8
Study Endpoints.....................................................................................................................15
9
Study Design..........................................................................................................................16
10
Treatment...............................................................................................................................19
11
Procedures Involved ..............................................................................................................25
12
Withdrawal of Subjects..........................................................................................................28
13
Risks to Subjects....................................................................................................................29
14
Potential Benefits to Subjects ................................................................................................30
15
Data and Specimen Banking..................................................................................................30
16
Measurement of Effect ..........................................................................................................30
17
Safety Evaluation...................................................................................................................31
18
Data Management and Confidentiality..................................................................................36
19
Statistical Plan .......................................................................................................................36
20
Provisions to Monitor the Data to Ensure the Safety of Subjects..........................................40
21
Vulnerable Populations..........................................................................................................40
22
Community-Based Participatory Research............................................................................40
23
Sharing of Results with Subjects ...........................................................................................40
24
Setting....................................................................................................................................40
25
Provisions to Protect the Privacy Interests of Subjects .........................................................40
26
Resources Available ..............................................................................................................41
27
Prior Approvals......................................................................................................................41
28
Compensation for Research-Related Injury...........................................................................41
29
Economic Burden to Subjects................................................................................................41
30
Consent Process.....................................................................................................................41
31
Process to Document Consent in Writing..............................................................................42
32
Drugs or Devices ...................................................................................................................42
33
References..............................................................................................................................46
34
Appendices/ Supplements......................................................................................................52
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1
STUDY OBJECTIVES
1.1
Primary Objectives

To determine the safety of the combination of IV. rintatolimod administered with or 
without IV IFNα (INTRON® A) in patients with cancer with COVID-19.

Determine the kinetics of viral load in nasopharyngeal swabs in the course of treatment 
and Days 7 and 14.
1.2
Secondary Objectives

To assess the efficacy of the treatment combination in patients with cancer with COVID-
19.

Determine the kinetics of viral load in the peripheral blood in the course of treatment and 
Days 7 and 14.

Determine the kinetics of changes of the immune subsets and circulating inflammatory 
mediators (including CRP, cytokines, chemokines, interferons) in peripheral blood in the 
course of treatment and Days 7 and 14.

Determine the induction of known mediators of antiviral immunity that include (myxovirus 
resistance gene, MxA; protein Kinase R (PKR); oligoadenylate synthetase-2  (OAS2); 
RNAse-L,  IFN-stimulated gene-15 (ISG15);  IFN-induced proteins with tetratricopeptide 
repeats (IFIT1) and IFN-inducible transmembrane protein 3 (IFITM3), TLR3, RIG-I, 
MDA5, IRF3, IRF7,  in nasopharyngeal swabs material and blood cells of patients on all 
tiers of treatment. Expression of ACE2 (receptor for SARS-Cov-2 entry) and potentially 
other genes involved in SARS-CoV-2 infection will be tested in nasopharyngeal samples.
2
BACKGROUND
2.1
Enhanced Risk of  COVID-19 Mortality and Morbidity in Cancer Patients
Patients with cancer and COVID-19 had a 5-fold higher risk of severe events, defined as ICU 
admission requiring invasive ventilation or death, than those without cancer (1). Currently, there 
is no proven effective treatment of COVID-19 beyond supportive care. This trial is based on the 
central premise that SARS-coronaviruses persist in host cells and in infected hosts by evasion 
of the innate antiviral responses mediated by RNAses (especially RNAse-L/OAS) and type-1 
interferon (IFN) pathway and IFN-stimulated genes (ISG), and that early therapeutic 
activation of this pathway will limit viral replication and expansion in patients’ tissues, thus 
preventing severe complications. SARS-Cov-2 is related to SARS, MERS, and other 
coronaviruses, which are positive-sense RNA viruses that generate dsRNA during replication. One 
of the principal IFN antiviral pathways involves activation of the host RNAses, especially RNase 
L, which is induced directly by dsRNA (predominantly by RIG-I and MDA5 helicases (2)) and 
indirectly by IFN degradation of viral RNA (3) and amplified IFN production (2). 
Coronaviruses avoid upstream pathways that activate type 1 IFNs by cytosolic pattern recognition 
receptors (e.g. RIG-I and MDA5) preferentially activated by the coronavirus RNA (2, 4), and 
inhibition or avoidance of IRF3, a transcriptional factor that mediates the induction of type 1 IFNs, 
by that pathway (5-9). This inhibition of innate immune responses allows the virus to replicate in 
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host cells, and particularly in high risk patients, progress to pneumonia and respiratory failure. To 
overcome this barrier to effective immunity, we will evaluate combination rIFNα and rintatolimod. 
Rintatolimod, a selective dsRNA ligand of TLR3 (but not RIG-I/MDA5 (10)), has strong antiviral 
activity against multiple viruses, and was found to be protective in a lethal mouse model of SARS-
CoV in vivo, and show strong anti SARS-CoV activity in vitro (11, 12). Combination rIFNα 
(INTRON® A; 20 MU/M2 per infusion) and rintatolimod (200 mg per infusion), which showed 
synergy in activating IFN response signature in preclinical models (10) is being evaluated in 
clinical trials developed by Dr. Kalinski and colleagues for patients with multiple solid tumors 
(active Roswell-held IND-112532 and trials NCT01545141, NCT03403634; NCT03403634), 
and, in over 70 enrolled patients, has not reached an MTD.
The rationale for the combination therapy in cancer is based on strong synergy in inducing ISGs 
and other IFN-regulated genes and promoting type-1 immunity (10); activation of these same 
pathways is expected to augment antiviral defense in patients with COVID-19. While rintatolimod 
activates the TLR3/IRF3 pathway to induce IFNα, it does not activate the MAVS/RIGI/MDA5, 
the alternative RNA-virus-sensing pathway preferentially activated by the coronavirus RNA (2, 
4), thus avoiding NF-kB and downstream cytokines (10) that could worsen acute lung injury in 
COVID-19. We expect INTRON® A to directly enhance the expression of OAS and its ability to 
promote the RNase L-mediated degradation of viral RNA in virally infected cells that do not 
mobilize own IFNα in response to COVID-19, while rintatolimod will induce TLR3-dependent 
ISGs in the uninfected bystander cells (10), and potentially infected cells, jointly leading to viral 
degradation. Augmentation of innate immunity in patients with COVID-19 is expected to avert 
viral progression to respiratory failure and mortality.
These results point to patients with cancer being at a several-fold higher risk for COVID-19-driven 
morbidity and mortality. Though many treatment modalities are being used, including single agent 
and combination antiviral agents and immunotherapy, there is no treatment proven to be effective 
for COVID-19 above supportive care. One promising option was use of HIV protease inhibitors 
for COVID-19; however, a recent trial showed lack of efficacy (13). Given the lack of effective 
therapies, there is an urgent need for more effective regimens that could be used prophylactically 
to prevent COVID-19 (e.g., vaccination) or as upfront therapy; this need is particularly pressing in 
patients with cancer. In addition, because there is no regimen shown to be more effective than 
supportive care, we did not include a comparator regimen.
2.2
COVID-19: Basic Biology and Rationale for Its Treatment Using the Rintatolimod 
(Ampligen®) and IFNα Combination
The COVID-19 is a coronavirus related to SARS, MERS, and other coronaviruses, which use RNA 
as their genetic material. It spreads in the body and in population by limiting early production of 
IFN and IFN-related genes, thus avoiding induction of host intracellular RNAse L (RNA-
degrading enzyme that degrades cytoplasmic host and viral RNA) and activation of innate 
immunity. There is a large body of literature showing that coronaviruses inhibit upstream pathways 
that activate type 1 interferons, including inhibition of intracellular signaling driven by pattern 
recognition receptors (e.g. RIG-I and MDA5) that sense viral RNA and by inhibition of IRF-3, a 
transcriptional factor that induces the expression of type 1 interferons (5-9). This inhibition of 
innate immune responses is expected to allow the virus to replicate in epithelial cells, and 
particularly in high risk patients, progress to pneumonia and respiratory failure. Our goal is to use 
INTRON® A to directly activate RNAse L in virally infected cells with defective ability to 
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generate endogenous interferon-alpha, while rintatolimod, through ligation of TLR3, will 
stimulate interferon-alpha expression in bystander uninfected cells, thereby protecting them from 
viral infection. We also expect the synergy between these two factors in activating IFN-inducible 
genes, similar to our observations in cancer (10), amplifying the impact of IFNα in infected cells. 
Seen in this light, augmentation of innate immunity during early mild or moderately severe 
infection might avert progression to respiratory failure and mortality. In patients with more severe 
COVID-19, including those with respiratory failure, rintatolimod may promote viral clearance and 
avert worsening of lung injury. However, augmentation of innate immune responses may also 
worsen inflammatory lung injury in patients with COVID-19 infection, and therefore safety and 
efficacy are primary endpoints of this trial. The current proposal involves repurposing of an 
investigational immunotherapy regimen developed by Dr. Kalinski for cancer (and currently 
being evaluated in several clinical trials) to be tested in patients with cancer and COVID-19 
disease or significant exposure.
Interferon-alpha (INTRON® A) and rintatolimod [Ampligen®] are two anti-viral drugs and 
immune activators, currently tested in combination in our ongoing the IRB- and FDA approved 
clinical trial I 52917 for patients with metastatic colorectal cancer and several other IRB- and 
FDA-approved trials in other cancers, performed at Roswell and the University of Pittsburgh.
Rintatolimod (selective TLR3 ligand, which activates multiple forms of antiviral immunity) has 
demonstrated strong antiviral activity against multiple viruses (see the attached information from 
AIM) including the coronavirus SARS COV-1 in vitro and in NIH-contracted animal experiments, 
conferring 100% protective survival. In vitro activity of Ampligen® against the human 
coronavirus, OC43, was performed by Barnard, D.L., et al. Proceedings of the 16th International 
Conference on Antiviral Research, 2003, using BS-C-1 kidney cells (African green monkey). In 
this in vitro test Ampligen® showed an EC50 of 0.4 μg/ml compared to the easily achievable 
concentration in humans of 40 μg/ml, which is well tolerated.  In in vivo studies (11) and studies 
at NIH of SARS-COV-1 infected mice (12), Ampligen® conferred conferring a significant 
antiviral/survival effect.
Since COVID-19 shares key genomic and pathogenic similarities with SARS-COV-1 viruses (see 
the attached information from AIM), it is also expected to be susceptible to rintatolimod.
The second drug, IFNα has shown antiviral activity against multiple types of viruses, including 
other coronaviruses, and is currently being tested as an anti-COVID-19 agent in multiple 
countries. 
The rationale for the combination of these two drugs in our ongoing clinical trials in cancer and 
in our proposed trial for COVID-19 patients results from their strong synergy in inducing IFN-
regulated genes and promoting type-1 immunity (similarly needed for anti-viral and anti-cancer 
immunity) in human tissues ex vivo (10) and the activity of this combination observed in 
experimental animals (14), as well as the very good safety record of this combination observed in 
cancer patients so far (see below).
2.3
Past experience with IFNα and Rintatolimod
Interferon–alpha (INTRON® A) is currently FDA approved for the adjuvant treatment of stage 
III melanoma. It has been examined by multiple investigators as an immune modulator in the 
setting of vaccine therapy with demonstrated safety (15). It has been extensively evaluated in the 
setting of metastatic CRC (as single agent or in combination with chemotherapy or IL-2), but was 
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shown to be largely ineffective (16-33) at daily doses ranging from 3-20 million units/m2, 
administered subcutaneously or intravenously. In the treatment of melanoma, a small study by 
Astsaturov and colleagues (34) administered IFNb in previously vaccinated melanoma 
patients, and was shown to convert immunization into objective clinical responses, suggesting its 
ability to amplify the effectiveness of local T cells infiltration and/or function at the tumor site. 
Vaishampayan et al. examined IFNα-2b 5 million units subcutaneously three times a week 
following vaccination with a melanoma vaccine (Melacine) (35). Treatment at this dose was well 
tolerated. Similarly Mitchell et al. recently completed a large trial examining IFNα-2b, 5 million 
units subcutaneously three times a week for one year (36). Di Pucchi and Pilla similarly examined 
the combination of a melanoma peptide vaccine with IFNα-2b, 3 million units subcutaneously 
three times a week, with little to no toxicity.
We propose to include a short course of high dose interferon alpha-2b (with the dose-escalation 
lead-in [0-5-10-20] and with maximum 20 million units/m2, IV, on Days 1 and 3). The maximum 
dose of 20 million units/m2 is similar or lower than the previously evaluated doses in melanoma 
patients (37-39). In this group of patients, in accordance with previous reports, we expect to 
consistently observe flu-like symptoms. However, this regimen hasn’t been evaluated in patients 
with respiratory viral infection, and additional AEs may be observed. Specifically, augmentation 
of innate immune responses may also worsen inflammatory lung injury in patients with COVID-
19 infection, and therefore safety and efficacy are primary endpoints of this trial.
Rintatolimod (Selective toll-like receptor-3 [TLR3] Ligand; analog of poly IC with reduced 
toxicity for i. v. use).
Rintatolimod (Ampligen®) is a TLR3 specific Poly IC analog in which the cytidylic acid chain has 
uridylic acid substitutions at a molar ratio of 12:1. Chemokine modulation in temporal association 
with type I interferon administration is observed with this agent. Rintatolimod has been studied 
extensively in multiple clinical settings including cancer, chronic viral infection, vaccination 
protocols, and in chronic treatment of chronic fatigue syndrome.
Rintatolimod has been generally well tolerated with only a low incidence of clinical toxicity. 
Clinical experience with rintatolimod now totals over 800 patients with more than 200 patients 
receiving rintatolimod for up to one (1) year, over 50 patients up to two (2) years, and more than 
20 patients over two (2) years.
The proposed dosage of 200 mg IV is the starting dose used in chronic treatment of Chronic 
Fatigue Syndrome (CFS). That dose is escalated to 400 mg IV twice weekly for the CFS indication; 
however, for use as a chemokine modulator, pharmacologic peak serum levels are achieved at 200 
mg; thus, 200 mg daily for 3 days will be incorporated for the immune modulation regimen. 
Preclinical evidence demonstrates the synergy between IFNα and rintatolimod in inducing the 
production of the effector T cell-attracting chemokines.
2.4
Clinical experience with Rintatolimod (Ampligen®) in Combination with Interferon 
Alpha-2b and Celecoxib
NCT01545141 (UPCI 10-131) was a Phase I study which evaluated the combination of 
rintatolimod, interferon-alpha 2b, and celecoxib administered for one course over 5 days, prior to 
tumor resection. The Phase I study was completed, with escalation to the maximal planned 
dose, without the emergence of undue toxicity. A 5-day regimen consisting of rintatolimod 200 
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mg IV daily, interferon alpha-2b (20 million units/ m2) IV daily and celecoxib 200 mg po bid was 
deemed highly tolerable and suitable for further investigation.
In addition to the demonstrated safety, preliminary evidence of biologic efficacy was observed. 
Individuals who were treated with CKM prior to resection, demonstrated an improvement in 
intratumoral CD8/FOXP3 ratios as well as CXCL10/CCL22 ratios when compared to 
nonrandomized controls, treated with standard care at the University of Pittsburgh (Zureikat, 
Bartlett and Kalinski, manuscript in preparation). The design (neo-adjuvant treatment) and 
numbers of patients treated do not permit any meaningful estimate of clinical outcomes or 
radiographic response at this time.
In addition to the evaluation of rintatolimod, interferon alpha-2b administered i. v. (combined with 
oral Celecoxib) for one course over 5 days, as a stand-alone treatment, the same combinations (but 
different administration regimens) are being evaluated as a part of combinatorial regimen (with 
DC vaccines) in additional patients with peritoneal carcinomatosis (NCT02151448 / UPCI 12-
110). Clinical trial NCT02151448 (over 50 patients evaluated so far) involve multiple cycles of 
DC vaccines followed by 4-day long cycles of IFN-a/celecoxib regimen, with rintatolimod 
administered on Days 2 and 4 of each cycle).
No DLTs have been observed in either of these trials, and we consider the INTRON® A dose of 
20MU/M2 as recommended i.v. dose for combination with i.v. rintatolimod. These 
concentrations of INTRON® A (20MU/M2) plus rintatolimod (200 mg) are being used in 
combination in our ongoing trials at Roswell Park in colon and breast cancer and in a recently 
approved trial in prostate cancer:

I 52917 (NCT03403634; IND 112532) Phase 2A Study Evaluating a Chemokine-
Modulatory Regimen in Patients with Colorectal Cancer Metastatic to the Liver. (Sarbajit 
Mukherjee and P. Kalinski)
Status:  COMPLETED; MANUSCRIPT IN PREPARATION 
Phase: Phase IIa (combination with Celecoxib). 

I 62218 (NCT03403634) Chemokine Modulation Therapy and Pembrolizumab in Treating 
Participants with Metastatic Triple-Negative Breast Cancer. (Shipra Gandhi and P. 
Kalinski) 
Status:  CLOSED TO ACCRUAL 
Phase: Phase IIa (combination with Celecoxib and Pembrolizumab). 

I 73718 Phase I Clinical Trial Assessing the Combination of Chemokine Modulation with 
Neoadjuvant Chemotherapy in Triple Negative Breast Cancer IND 145344). (Shipra 
Gandhi and P. Kalinski)
Status:  COMPLETED; MANUSCRIPT IN PREPARATION 
Phase: Phase I (combination with Celecoxib and Paclitaxel). 

I 77318 (NCT03899987) Aspirin and rintatolimod With or Without Interferon-alpha 2b in 
Treating Patients with Prostate Cancer Before Surgery” (Gurkamal Chatta and Pawel 
Kalinski)
Status: FDA (IND 143104) and IRB approved. ACCRUING PATIENTS
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2.5
Rationale for eliminating celecoxib from the proposed regimen and for reducing, 
eliminating, or changing the source and route of administration of IFNα
In view of the potential concerns regarding the use of NSAIDs in patients with COVID-19, we 
decided not to include Celecoxib in the current trial.
The dose of INTRON® A IFNα may be reduced from 20MU/m2 to 5 MU/m2 or to 0), in case of 
any observed toxicities, or in case of IFNα shortage.
In case of potential shortages, INTRON® A may be replaced by another form of IFNα at the same 
(Units) dose. In case the only available IFNα is formulated for an alternative route of 
administration (e.g., subcutaneous), we will adjust the dose to use the most common dose for that 
specific product. An IRB amendment will be submitted for any changes in formulation or dosing.
2.6
Overall Impact
Our study focuses on patients with cancer, but knowledge gained about safety and immunologic 
correlates is expected to be applicable to other patients at high risk for COVID-19-related 
morbidity and mortality as well as to future viral pandemics. Using each patient’s baseline 
nasopharyngeal viral load, we will evaluate to what extent the study regimen reduces viral load 
relative to supportive care only. Assessment of nasopharyngeal viral load at multiple time points 
will enable evaluation of early and long-term effects of the study regimen in controlling viral 
replication. A reduction in nasopharyngeal viral load by PCR may also reflect reduced likelihood 
of transmission of virus to others. As described in the Statistical Plan (Section 19), the study is 
non-randomized and each patient will serve as their own control regarding baseline, on-treatment, 
and post-treatment nasopharyngeal viral load. In addition, comparisons will be made with 
historical controls in which Roswell Park patients with COVID-19 were treated with standard of 
care and longitudinal viral load was monitored as standard of care. As described in the 
demonstration of safety of this regimen and reduction of nasopharyngeal viral load will establish 
the foundation for a larger randomized trial powered for clinical efficacy, such as reduction in 
respiratory failure and mortality. 
Our study will enroll patients with cancer with varying comorbidities and levels of COVID-19 
severity. For outpatients with COVID-19 at high risk for severe complications based on age and 
co-morbidities, a monoclonal antibody cocktail (e.g. Regeneron’s combination of casirivimab and 
imdevimab) is standard of care. In patients with more severe COVID-19 who are hospitalized and 
require supplemental oxygen, therapy is focused on controlling inflammatory injury. 
Dexamethasone was associated with a survival benefit and will be administered to all patients who 
meet criteria for this agent. Other anti-inflammatory agents such as IL-6-targeting agents and 
baricitinib can also be of value, although their benefit when added to corticosteroids is unclear. 
The antiviral agent, remdesivir, is of unclear value based on conflicting data; at best it shortens the 
duration of illness but doesn’t have a survival benefit. At Roswell, we routinely administer 
combination dexamethasone and remdesivir to inpatients with significant COVID-19, and will do 
so for patients enrolled in this protocol unless there is a clinical contraindication to receiving either 
agent. The data on convalescent plasma is also contradictory, but plasma with high neutralizing 
titers against the viral Spike protein is likely to be of value. We do not know the effect of these 
standard of care regimens on nasopharyngeal viral load in patients with cancer. In addition, since 
these regimens are often administered concurrently, it’s difficult to delineate the effect of a specific 
drug on viral load. Based on our experience at Roswell Park, the nasopharyngeal viral load in some 
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immunocompromised patients with cancer (e.g. stem cell transplant recipients and patients 
receiving B-cell-depleting agents), has persisted for several weeks despite receiving combinations 
of these agents (e.g. remdesivir and dexamethasone, often in combination with convalescent 
plasma). The standard of care for COVID-19 changes over time. Regarding the current protocol, 
the study regimen will be added to what in our judgment is the best standard of care regardless of 
outpatient or inpatient status.
3
INCLUSION AND EXCLUSION CRITERIA
3.1
Inclusion Criteria (Main Cohort)
To be included in this study, participants must meet the following criteria:
1
Patients with cancer, with the exception of patients with active acute leukemia and 
allogeneic hematopoietic stem cell transplant recipients. Patients may be  on active therapy 
or received therapy (e.g., chemotherapy, radiation, or surgery) within 7 years. Patients with 
active cancer who have not yet been treated (e.g. newly diagnosed cancer or early stage 
MDS or CLL) are eligible. Basal cell cancer and carcinoma in situ treated with local 
excision alone do not qualify for inclusion.
2
Presence of symptomatic infection, defined by fever (T≥38.0°C ) OR respiratory symptoms 
(cough, nasal congestion, or shortness of breath), OR lung infiltrates on chest X-ray or CT 
imaging, Diagnosis of COVID-19 is based on PCR testing of respiratory samples. 
3
Age equal to ≥ 18 years or older (children are excluded because COVID-19 typically has 
a milder course in children, and lack of safety data of this regimen in children).
4
Laboratory evaluation:
o
Platelet ≥ 75,000/µL
o
Hemoglobin ≥ 9 g/dL
o
Hematocrit ≥ 27%
o
Absolute Neutrophil Count (ANC) ≥ 1000/µL
o
Creatinine clearance ≥ 50 mL/min (Cockroft-Gault Equation–note: plasma creatine 
instead of serum is used at Roswell Park)
o
Total bilirubin ≤ 2 X institutional ULN 
o
AST (plasma) and ALT(plasma) ≤ 2 X institutional ULN
o
Plasma amylase and lipase ≤2 X institutional ULN
5
In the absence of COVID-19, a life expectancy of 6 months is expected.
6
Participant must understand the investigational nature of this study and sign an Independent 
Ethics Committee/Institutional Review Board approved written informed consent form 
prior to receiving any study related procedure.
NOTE: For blood chemistry labs, Roswell Park clinical blood chemistries are performed on 
plasma unless otherwise indicated.
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Refer to Appendix D for the Investigator Study Eligibility Verification Form (Main Cohort): 
Inclusion Criteria.
3.2
Exclusion Criteria (Main Cohort)
Participants will be excluded from the study for the following:
1
Patients with severe COVID-19 infection defined by pulmonary infiltrates on Chest X-Ray 
or CT imaging plus one of the following: room air SaO2 ≤ 92%, room air PaO2 <70 mm 
Hg, or PaO2-PaO2 ≥35 mm Hg.
2
Contraindication to r-IFNα based on prior hypersensitivity, autoimmune hepatitis, 
decompensated liver disease.
3
Patients who have active acute myeloid leukemia or acute lymphoid leukemia or are 
allogeneic hematopoietic stem transplant recipients. Acute leukemia in remission and 
chronic leukemias are not exclusion criteria.
4
Cardiac events:
a.
Acute coronary syndrome, myocardial infarction, or ischemia within past 3 
months,
b. New York Heart Association classification of III or IV congestive heart failure 
(Appendix A)
5
Unwilling or unable to follow protocol requirements.
6
Patients with known serious mood disorders.
7
Any additional condition, such as pre-existing inflammatory lung disease, which in the 
Investigator’s opinion deems the participant an unsuitable candidate to receive the study 
drugs.
8
Concurrent infections, e.g. bacterial pneumonia or sepsis, that would make it difficult to 
evaluate clinical response to therapy or study drug toxicities.
9
Therapies known to cause cytokine release syndrome (CRS), e.g. engineered T cells, within 
30 days.
10 Patients at high risk for tumor lysis syndrome.
11 Concurrent active pneumonitis predating COVID-19, such as from checkpoint inhibitor 
therapy, chemotherapy-associated toxicity, or radiation pneumonitis.
12 Autoimmune disease that requires systemic immunosuppression.
13 Protocol-defined baseline abnormalities in cell counts, renal, or hepatic function.
14 Any additional condition which in the Investigator’s opinion deems the participant an 
unsuitable candidate to receive the study drugs.
Refer to Appendix E for the Investigator Study Eligibility Verification Form (Main Cohort): 
Exclusion Criteria.
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3.3
Inclusion Criteria (Expansion Cohort: Rintatolimod Plus Best Available Standard 
Care)
We have added an expansion cohort to receive rintatolimod plus best available standard care. 
Because rIFN-α is not included in this group, eligibility criteria have been liberalized to include 
patients with more severe COVID-19, including those with respiratory failure,  and exclusion 
criteria specific to rIFN-α have been removed. For patients who are eligible for both the main 
cohort and expansion cohort, enrollment in the Main Cohort will be prioritized because of the 
rationale that it might be more effective. To be included in the Expansion Cohort, cancer 
participants must meet the following criteria:
1
Patients with cancer or allogeneic stem cell transplant recipients with and without a cancer 
diagnosis. 
o
Patients with cancer may be on active therapy or received therapy (e.g., 
chemotherapy, radiation, or surgery) within 7 years. 
o
Patients with active cancer who have not yet been treated (e.g. newly diagnosed 
cancer or early stage MDS or CLL) are eligible.  
o
Basal cell cancer and carcinoma in situ treated with local excision alone do not 
qualify for inclusion.
2
Presence of symptomatic infection, defined by fever (T≥38.0°C ) OR respiratory symptoms 
(cough, nasal congestion, or shortness of breath) OR lung infiltrates by chest X-ray or CT 
imaging.  Diagnosis of COVID-19 is based on PCR testing of respiratory samples. Severe 
infection is excluded (see Exclusion Criteria).
3
Age equal to ≥ 18 years or older (children are excluded because COVID-19 typically has 
a milder course in children, and lack of safety data of this regimen in children). In the 
absence of COVID-19, a life expectancy of 6 months is expected.
4
Participant or health care proxy must understand the investigational nature of this study 
and sign an Independent Ethics Committee/Institutional Review Board approved written 
informed consent form prior to receiving any study related procedure. There may be 
specific instances when the patient can’t provide informed consent, e.g. they require 
mechanical ventilation and are sedated, in which case a health care proxy will be able to 
provide informed consent. Patients with temporary cognitive impairment will be consented 
once their capacity has returned. Patients with chronic cognitive impairment, e.g. dementia, 
that precludes informed consent will not be enrolled.
NOTE: For blood chemistry labs, Roswell Park clinical blood chemistries are performed on 
plasma unless otherwise indicated.
Refer to Appendix D for the Investigator Study Eligibility Verification Form (Expansion Cohort): 
Inclusion Criteria.
3.4
Exclusion Criteria (Expansion Cohort)
Participants will be excluded from the study for the following:
1
Patients with respiratory failure requiring mechanical ventilation with FIO2 of > 60%. 
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2
Allogeneic hematopoietic stem cell transplant recipients with active pulmonary GvHD 
(any grade). 
3
Cardiac events: 
a.
Acute coronary syndrome, myocardial infarction, or ischemia within past 3 
months,
b. New York Heart Association classification of III or IV congestive heart failure 
(Appendix A)
4
Unwilling or unable to follow protocol requirements.
5
Any additional condition which in the Investigator’s opinion deems the participant an 
unsuitable candidate to receive the study drugs.
Refer to Appendix E for the Investigator Study Eligibility Verification Form (Expansion Cohort): 
Exclusion Criteria.
3.5
Inclusion of Women and Minorities
Both men and women and members of all races and ethnic groups are eligible for this study.
3.6
Special Populations
The following special populations will be excluded:

Chronically cognitively impaired adults (e.g. those with dementia) with impaired decision-
making capacity

Individuals who are not yet adults (infants, children, teenagers)

Pregnant women

Prisoners
4
LOCAL AND STUDY-WIDE NUMBER OF SUBJECTS
Maximum number of Roswell Park subjects: The anticipated number of subjects required for the 
main portion of  the study is 44. All patients must have cancer and need not be established Roswell 
Park patients. The modification of the protocol triggered by the observed high effectiveness of the 
treatment in the first patient who received rintatolimod alone will include additional 20 cancer 
patients (Expansion Cohort). 
4.1
Target Accrual
Main Cohort: Up to 44 patients are expected to be needed to estimate the biologic impact of this 
regimen and establish a preliminary estimate of efficacy. This number includes the following tiers 
of the dose-escalation of IFNα (3+3 design with 0, 5 MU/M2, 10 MU/M2, and 20 MU/M2). Once 
an MTD is established, subsequent patients (n = 10) will receive rintatolimod +  IFNα (at the MTD 
or, in the absence of an MTD, at 20 MU/M2 IFNα) as well as standard of care. Total accrual time 
is expected to take 36 months. 
Expansion Cohort: In addition, we will accrue a maximum of 20 patients on Expansion Cohort, 
who will receive  rintatolimod and best available standard care. 
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5
LOCAL AND STUDY-WIDE RECRUITMENT METHODS
5.1
Recruitment
Subjects will be recruited from Roswell Park and our affiliate practices, and University at Buffalo-
affiliated hospital systems. Recruitment efforts will focus on patients with initial signs of COVID-
19 who would be offered enrollment as soon as feasible. In addition, asymptomatic patients with 
contact (e.g., household member or health care-associated exposure) with COVID-19 will be 
monitored for development of symptoms or signs of COVID-19 that would result in 
nasopharyngeal swab PCR testing: Roswell Park is a New York State-designated regional 
laboratory for COVID-19 PCR-based testing with same or next day reporting of results. 
Men and women will be recruited, with age, race, and socioeconomic status expected to reflect the 
general Roswell Park patient population.
6
MULTI-SITE RESEARCH
The study will be opened at Roswell Park first to obtain preliminary safety data and establish 
feasibility, but we may prospectively partner with other cancer centers as study sites in regions 
with established community-based transmission of COVID-19.
7
STUDY TIMELINES
Main Cohort: A maximum of 44 evaluable participants will be enrolled to the study over up to 36-
months. Patients will be followed until Day 30 after initiation of the study regimen.
Expansion Cohort: A maximum of 20 evaluable participants will be enrolled to the study over up 
to 36-months. Patients will be followed until Day 30 after initiation of the study regimen. 
Enrollment in the Main Cohort and Expansion Cohort will be done concurrently. If a patient is 
eligible for enrollment in both the Main and Expansion Cohort and is willing to be enrolled in 
either cohort, priority will be given to the Main Cohort.
8
STUDY ENDPOINTS
8.1
Primary Endpoints

The primary safety endpoints are frequency of grade 3 or 4 AEs considered to be probably 
or definitely related to the treatment regimen.

Evaluate kinetics of viral load expressed as Ct values in nasopharyngeal swab samples in 
the course of treatment and Days 7 and 14. (40)
8.2
Secondary Endpoints*

Clinical efficacy will be assessed by the frequency of these complications: (i) progression 
of infection requiring hospitalization; (ii) respiratory failure requiring mechanical 
ventilation; and (iii) death within 30 days.
Although the data on outcomes in patients with COVID-19 and cancer are limited, based on the 
Chinese experience, approximately 40% are expected to have the composite endpoint of 
respiratory failure requiring mechanical ventilation or death (1). We note the limitations of this 
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estimate regarding small numbers of patients with cancer and heterogeneity in cancer types and 
co-morbidities, including older age, smoking, and lung disease (41). If present, acute respiratory 
distress syndrome (ARDS) will be graded by Berlin criteria (42), which include 3 mutually 
exclusive categories of ARDS based on degree of hypoxemia: mild (200 mm Hg < PaO2/FIO2 ≤ 
300 mm Hg), moderate (100 mm Hg < PaO2/FIO2 ≤ 200 mm Hg), and severe (PaO2/FIO2 ≤ 100 
mm Hg). The degree of ARDS is has been validated as a predictor of mortality in patients with 
respiratory failure. The Berlin criteria require 5cm H2O as the minimum level of PEEP delivered 
by a ventilator or CPAP for diagnosing ARDS; if PEEP is not administered, then ARDS cannot be 
diagnosed. In addition, the Berlin criteria require excluding hydrostatic pulmonary edema by 
clinical judgement or with echocardiograph as a cause of respiratory failure. Although the study is 
not powered for the clinical endpoint, in case that any of the elements of the clinical efficacy 
endpoint will reach statistical difference between arms, such clinical endpoint will be considered 
as an overruling endpoint of efficacy

Evaluate kinetics of viral load as Ct values in peripheral blood in the course of treatment 
and Days 7 and 14 (40).

Determine the kinetics of changes of the immune subsets and circulating inflammatory 
mediators (including CRP, cytokines, chemokines, interferons) in peripheral blood in the 
course of treatment. Plasma and PBMCs will be cryopreserved for future exploratory 
studies, including the assessment of intracellular signaling and TCR and BCR repertoires 
as correlates of response to the study regimen.

Determine the induction of known mediators of antiviral immunity that include (myxovirus 
resistance gene, MxA; protein Kinase R (PKR); oligoadenylate synthetase-2  (OAS2); 
RNAse-L,  IFN-stimulated gene-15 (ISG15);  IFN-induced proteins with tetratricopeptide 
repeats (IFIT1) and IFN-inducible transmembrane protein 3 (IFITM3), TLR3, RIG-I, 
MDA5, IRF3, IRF7,  in nasopharyngeal swab  material and blood cells of patients on all 
tiers of treatment. Expression of ACE2 (receptor for SARS-Cov-2 entry) and potentially 
other genes involved in SARS-CoV-2 infection will be tested in nasopharyngeal samples.
9
STUDY DESIGN
The antiviral efficacy of the combination of interferon alpha-2b and rintatolimod will be evaluated 
by administering fixed doses of each drug. Subjects will receive 1 day of the combination (Day 1), 
followed by 1 day of observation and the same regimen on Day 3 (or Day 4). according to the safe 
established dosing levels (see Table 1). During this time, subjects will be monitored continuously 
for safety (refer to Section 19.2.1). As the standard of care therapy for COVID-19 is evolving, all 
patients will receive optimal standard of care that will be administered concurrently with the study 
drug regimen. In a double-blind, randomized, multi-center, placebo-controlled trial of intravenous 
remdesivir in adults hospitalized with COVID-19 with evidence of lower respiratory tract 
involvement, remdesivir treatment resulted in shortening the time to recovery by 4 days (43). 
Though 14-day survival was greater in remdesivir recipients, the difference was not statistically 
significant  (43). Adults hospitalized with COVID-19 with evidence of lower respiratory tract 
involvement will be offered remdesivir through an EUA if and when  this drug is available to us. 
If we’re not able to offer remdesivir to all patients based on drug availability, we expect a similar 
proportion of patients in the study drug and control arms to have received remdesivir, and the 
effect of remdesivir on study endpoints will be evaluated in an exploratory fashion.
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In a multi-center controlled, open-label trial comparing a range of possible treatments in patients 
who were hospitalized with COVID-19, patients were randomly assigned to receive oral or 
intravenous dexamethasone (6 mg daily) for up to 10 days or to receive usual care alone (44). 
Overall, 482 patients (22.9%) in the dexamethasone group and 1110 patients (25.7%) in the usual 
care group died within 28 days after randomization (p < 0.001). The survival benefit of 
dexamethasone was observed among patients receiving either invasive mechanical ventilation or 
oxygen alone at randomization but not among those receiving no respiratory support. Based on 
these results, dexamethasone (6 mg/day) will be given as standard of care to patients admitted with 
COVID-19 who require supplemental oxygen.
Nasopharyngeal swab collection: Nasopharyngeal swabs will be performed by trained nursing 
staff under full PPE precautions. Nasopharyngeal swabs will be obtained using a single swab for 
both nostrils as specified in CDC guidelines: https://www.cdc.gov/coronavirus/2019-
ncov/lab/guidelines-clinical-specimens.html. Swabs are placed in viral media and submitted 
immediately on room air to the Dept. of Laboratory Medicine at Roswell Park. RT-PCR for 
SARS-CoV-2 testing is done under the direction of Dr. Jan Nowak (Professor of Oncology, 
Pathology, and Molecular Pathology). Testing for SARS-CoV-2 at Roswell Park received New 
York State (NYS) EUA. Roswell Park uses the New York SARS-CoV-2 Real-time RT-PCR 
Diagnostic Panel, which received EUA from the FDA. As a NYS-authorized laboratory for 
COVID-19 testing, Roswell Park’s diagnostic methods are as stipulated in the New York SARS-
CoV-2 Real-time RT-PCR Diagnostic Panel authorized procedures. The N1 and N2 PCR 
reactions target different regions of the SARS CoV-2 RNA. The New York SARS-CoV-2 Real-
time RT-PCR Diagnostic Panel requires the following control materials:

Human Specimen Control (HSC): A human cell culture preparation used as an extraction 
control and positive control for the RNase P primer and probe set that is extracted and 
tested concurrently with each specimen extraction run.

SARS-CoV-2 Positive Control (SARS-CoV-2 Pos): Run with each batch of specimens. 
Monitors improper assay setup, reagent failures of rRT-PCR reagents and reaction 
conditions.

No Template Control (NTC): Nuclease-free water included in each run. Monitors for 
reagent and system contamination.

RNase P (RP) control in clinical samples: The RP primer and probe set is included in each 
run to test for human RNase P, which controls for specimen quality and demonstrates that 
nucleic acid was generated by the extraction process.
Two PCR reactions using N1 and N2 primer sets target different regions of the SARS CoV-2 
RNA.  A specimen is considered positive if both N1 and N2 Ct values are < 40. Specimens with 
one value <40 and one >40 after repeat testing are interpreted as “Inconclusive”. Specimens which 
are negative show no amplification for both N1 or N2 PCR reactions. Based on our experience, 
N1 Ct values are typically several cycles lower than the N2 values, and therefore N1 Ct values will 
be used in statistical analyses.
*Validation 
studies 
included 
in 
the 
assay 
IFU 
(Instructions 
For 
Use) 
(https://www.fda.gov/media/134922/download) establish a Limit of Detection (LoD) of 25 
genome copies/reaction.  In defining the performance characteristics of the test, the IFU shows 
that this limit typically correlates with Ct values below 40 cycles. To assure test performance 
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consistency, positive target controls and extraction controls are required to show Ct values < 40 
cycles for test run validation.  Specimens with one value <40 and one >40 after repeat testing are 
interpreted as “Inconclusive”.
Table 1
Dosing
For the Main Cohort: Interferon 
alpha-2b dose-escalation IV over 20 
minutes on Day1 and on Day 3 or 4. 
The dose-escalation will use a 
standard 3 + 3 design, with the 
following doses: 0, 5, 10, and 20 
MU/M2.
Subsequent patients receive IFNα at 
20 MU/M2 or at MTD + 
rintatolimod (200 mg) on Day 1 and 
on Day 3 (or 4) as well as standard 
of care.
Initial administration to start at 20 mL/ hour and increase to 40 mL/ hour 
after 30 minutes as per the standard at Roswell. Total duration of 
rintatolimod infusion: 2.5 h-3h. Tubing should be flushed with 30 to 50 
mL of normal saline solution upon completion. Administration will be 
followed by 1 hour of observation and vital signs at 30 and 60 minutes 
post-infusion (± 5 minutes).
Figure 1
Study Schema (Main Cohort)
Patients will be seen for an end of treatment visit on Days 7 and 14  to review adverse events and 
clinical outcome of therapy.
Following the implementation of this protocol, we observed:
1. Difficulty to accrue patients with cancer and original inclusion criteria, as a result of strict 
inclusion criteria reflecting contra-indications for IFNα use.
2. The first patient treated at a lowest tier cleared the virus already on Day 3 of treatment 
before the administration of the second dose of the vaccine. 
In this situation, the current modification of the protocol includes an additional 20 cancer patients 
(Expansion Cohort), which will receive of rintatolimod as well as Standard of Care. The 
inclusion criteria are different than the main protocol. 
Enrollment
Day 1
Day 3
or
Day 4
Specimen collections (blood, nasopharyngeal swabs).
Day 7
(± 1 day)
Day 14
(± 3 days)
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10 TREATMENT
All patients will receive best available standard of care. Depending on the severity of COVID-19 
and need for hospitalization, standard of care can include, but is not limited to,  monoclonal 
antibody formulations directed against the SARS-CoV-2 Spike protein, remdesivir,  
dexamethasone, and tocilizumab. Treatment in the Main Cohort will consist of lead-in single agent 
rintatolimod (first 3 patients on the dose-escalation component) followed by the combination of 
rintatolimod and dose-escalation of rIFNα. Intravenous (IV) rintatolimod is administered as a fixed 
dose (200 mg per infusion) and IV rIFNα dose-escalation will be a standard 3 + 3 design, with the 
following doses per infusion: 0, 5, 10, and 20 MU/M2. The dose-escalation phase will lead to 
which patients will receive IFNα at 20 MU/M2 or at MTD + rintatolimod (200 mg). The regimen 
will be administered on Day 1 and on Day 3 (or 4), and EOT assessments will occur on Day 
7 (± 1 day) and Day 14 (± 3 days). This portion of the study will allow for additional safety data 
and the analysis of the effect of the study regimen on nasopharyngeal viral load. Using each 
patient’s baseline nasopharyngeal viral load, we will evaluate to what extent the study regimen 
reduces viral load relative to supportive care only.
Treatment in the Expansion Cohort will be IV rintatolimod is administered as a fixed dose (200 
mg per infusion on Day 1 and on Day 3 (or 4). Based on the PI’s judgment, additional infusions 
of  rintatolimod at 200 mg may be administered 3 to 4 days apart. Decisions about additional 
rintatolimod infusions will be based on evaluation of AEs attributed to rintatolimod, particularly 
increased FIO2 needs. A maximum of 12 rintatolimod infusions may be administered per patient. 
The rationale for a maximum of 12 infusions was based on discussions with leadership from AIM 
Immunotech regarding the expected effects of rintatolimod on innate immune responses over time 
based on data from patients with and without cancer. Rintatolimod activates innate immune 
responses through a number of mechanisms including activation of RNase L and inducing 
interferon stimulated genes (ISGs). Administration of rintatolimod over a number of weeks may 
be required for sustained activation of innate immune responses to clear SARS-CoV-2. 
10.1 Dosing and Administration
Interferon-Enhancing Regimen
For dosing, refer to Table 1. The regimen will be administered on Days 1 and 3 (or 4) in the 
following order:

Pretreatment: 500 mL Normal saline IV over 60 minutes

Pre-medications: acetaminophen (Tylenol®) 650 mg by mouth x 1 dose; prochlorperazine 
(Compazine®) 10 mg by mouth x 1 dose – administered 30 minutes (±5 minutes) after 
starting pre-treatment hydration

Interferon alpha-2b: (dose-escalation; maximum 20 MU/M2) IV over 20 minutes 
Rintatolimod: 200 mg IV, initial administration to start at 20 mL/ hour and increase to 40 mL/ hour 
after 30 minutes as per the standard at Roswell. Total duration of rintatolimod infusion: 2.5 h-3h. 
Tubing should be flushed with 30 to 50 mL of normal saline solution upon completion. 
Administration will be followed by 1 hour of observation and vital signs at 30 and 60 minutes 
post-infusion (± 5 minutes).
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Pretreatment and pre-meds listed above are recommendations and may be modified based upon 
institutional SOC requirements and at the discretion of the investigator. Treatment can be 
administered in the outpatient or inpatient setting, per the investigator’s/physician’s discretion.
Each daily treatment will be initiated after interval history and physical examination is performed 
and once the relevant laboratory studies have been confirmed to be within clinically acceptable 
ranges, specifically: ANC ≥ 1000/mm3, total bilirubin ≤ 3x ULN, AST and ALT ≤ 5 x ULN.
Expansion Cohort (Rintatolimod plus best available standard of care)
20 patients on Expansion Cohort will receive rintatolimod plus best available care.
Pretreatment for rintatolimod will be:

500 mL Normal saline IV over 60 minutes

Pre-medications: acetaminophen (Tylenol®) 650 mg by mouth x 1 dose; prochlorperazine 
(Compazine®) 10 mg by mouth x 1 dose – administered 30 minutes (±5 minutes) after 
starting pre-treatment hydration 
Rintatolimod: 200 mg IV, initial administration to start at 20 mL/ hour and increase to 40 mL/ hour 
after 30 minutes as per the standard at Roswell. Total duration of rintatolimod infusion: 2.5 h-3h. 
Tubing should be flushed with 30 to 50 mL of normal saline solution upon completion. 
Administration will be followed by 1 hour of observation and vital signs at 30 and 60 minutes 
post-infusion (± 5 minutes).
Pretreatment and pre-meds listed above are recommendations and may be modified based upon 
institutional SOC requirements and at the discretion of the investigator. Treatment can be 
administered in the outpatient or inpatient setting, per the investigator’s/physician’s discretion.
10.2 Dose Modification
Based on our experience with the combination of rintatolimod and INTRON® A as anti-cancer 
treatment (over longer periods of time than the currently proposed trial for COVID-19), we do not 
expect the need for dose modifications. However, in case of specific toxicities, dose modification 
rules will be used (Table 2). Taking into account that the rationale for the regimen is a short-term 
intervention to augment antiviral immunity for an acute viral infection, we do not plan for re-
escalations or administration of any missing doses after successful management of toxicities. Since 
the progression of COVID-19 is believed to be facilitated by lack of the activation of innate 
immunity (IFN responses and RNases), we anticipate that even transient (or lower intensity)  
activation of these pathways may help to better control the viral spread. Toxicity will be assessed 
according to the NCI Common Terminology Criteria for Adverse Events Version 5.0 (CTCAE 
v5.0).
Therapy will be immediately discontinued for any grade IV treatment-related toxicity that becomes 
apparent. In the event of any adverse effect, appropriate medical treatment will be instituted and 
study treatment will be discontinued based on physician judgment.
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Table 2
Treatment Related Toxicity Management
Treatment-
related AEs
Toxicity grade 
or conditions 
(CTCAE5.0)
Action taken for 
Interferon 
Alpha-2b
Action taken 
for 
Rintatolimod
Additional actions
Chills/Fever
Grade 3 that 
persists for ≥ 48 
hours despite 
acetaminophen
None
None
Consider additional acetaminophen 
dosing, supportive care
Pancreatitis
Grade 3
Hold
Decreased future 
doses by 75%
 (to 5MU/M2)
Hold
Supportive measures as clinically 
indicated
AST / ALT 
elevation or 
Increased 
bilirubin
Grade 3
Hold until 
improved to grade 
1; Decrease future 
doses by 75%
Hold
Advise evaluation for other possible 
etiologies; consider imaging
Vomiting
Grade 3 
vomiting ≥ 48 
hours despite 
optimal anti-
emetic therapy
Hold until 
resolved to grade 
1; Decrease future 
doses by 75% 
Hold
Depression or 
other clinically 
relevant mood 
disorder
Grade 3
Permanently 
discontinue; 
appropriate 
supportive 
care/intervention
None
Patients and family members will be 
informed in advance about the 
potential for depression and anxiety, 
including worsening of pre-existing 
mood disorders.
Neutropenia
Grade 3 or 4 
If Grade 3, 
decrease future 
doses by 75%. If 
Grade 4, 
permanently 
discontinue
Hold
Transient neutropenia is expected as 
a result of neutrophil activation and 
redistribution rather than bone 
marrow suppression. Recent 
cytotoxic chemotherapy is an 
additional cause of neutropenia. 
COVID-19 is known to cause 
lymphopenia, but not neutropenia. 
For prolonged neutropenia that is 
not expected from the study 
regimen and recent chemotherapy 
(if applicable), the primary 
oncologist and a hematologist will 
be consulted.
Hemorrhage
Grade 2 or 
greater
None
None
Myocardial 
infarction, stroke, 
Permanently 
discontinue; 
appropriate 
 Permanently 
discontinue; 
appropriate 
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Treatment-
related AEs
Toxicity grade 
or conditions 
(CTCAE5.0)
Action taken for 
Interferon 
Alpha-2b
Action taken 
for 
Rintatolimod
Additional actions
or other arterial 
thrombotic event
supportive 
care/intervention
supportive 
care/
intervention
Intolerable 
Grade 2 or 
Grade 3
Hold therapy until 
resolved to Grade 
1 or baseline; 
reduce future 
doses by 75%
Hold therapy 
until resolved 
to Grade 1 or 
baseline; 
reduce future 
doses by 50%
All other 
clinically 
significant AEs1
Grade 4
Permanently 
Discontinue
Permanently 
Discontinue
Cytokine 
Release 
Syndrome2
≥ Grade 2, 
based on the 
Penn scale (45)
Permanently 
Discontinue
Permanently 
Discontinue
In discussion with the primary 
oncologist, tocilizumab, 
corticosteroids, and potentially other 
immunomodulators will be used 
based on clinical judgement for 
ARDS and CRS-like complications.
1 Transient neutropenia is known to occur with rIFN-α and may reflect increased neutrophil migration to sites of 
infection or the tumor microenvironment as well as the effect of IFN-α on neutrophil survival and lifespan (46). This 
transient neutropenia has not been associated with an increased risk of infection.
2 Refer to: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5833070/  (Porter D, Frey N, Wood PA, Weng Y, 
Grupp SA. Grading of cytokine release syndrome associated with the CAR T cell therapy tisagenlecleucel 
[published correction appears in J Hematol Oncol. 2018 Jun 13;11(1):81]. J Hematol Oncol. 2018;11(1):35. 
Published 2018 Mar 2. doi:10.1186/s13045-018-0571-y).
10.2.1 Interferon Alpha-2b and/or rintatolimod Non-hematologic toxicities
Considered to be possibly, probably, or definitely related to interferon alpha-2b and rintatolimod:

Interferon Alpha-2b:
For grade 3 toxicity not addressed in Table 2:
o
1st episode: dose will be held and restarted at the next study visit at a dose reduction 
by 75%. The dose can be re-escalated if toxicity is reduced to ≤ grade 1 or baseline.
o
2nd episode: Treatment will be discontinued.
For grade 4 toxicity
o
Treatment will be discontinued
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
Rintatolimod:
For grade 3 toxicity not addressed in Table 2:
o
1st episode: dose will be held and restarted at the next study visit at a dose reduction 
by 50%. The dose can be re-escalated if toxicity is reduced to ≤ grade 1 or baseline.
o
2nd episode: Treatment will be discontinued.
For grade 4 toxicity:
o
Treatment will be discontinued
10.2.2 Definition of DLT
Treatment-related DLTs will be defined as the following events that occur between Day 1 to Day 
7 that are considered possibly, probably, or definitely treatment-related:

Grade 4 hematologic toxicity.

Grade 3 hematologic toxicity with exception of transient leukopenia, neutropenia, 
lymphopenia, thrombocytopenia without bleeding, or anemia lasting 7 days or less.

Grade 4 non-hematologic toxicities, with exception of asymptomatic increased LDH or 
lipase or amylase.

Grade 3 non-hematologic toxicities with exception of nausea, vomiting, diarrhea, or flu-
like symptoms that are controlled by optimal supportive management.
Distinguishing clinical manifestations of COVID-19 from treatment-related DLT can be difficult 
because they can overlap. Manifestations of COVID-19 present at baseline (e.g., fever, shortness 
of breath) that are unchanged after the study regimen is begun will not be considered DLT. 
Worsening of these manifestations can be due to COVID-19 or treatment-related, and unless there 
is a specific well defined unrelated cause, all AEs that are at least possibly related to the study 
regimen that meet one of the criteria for DLT based on CTCAE severity will be considered DLT.
In patients who received recent chemotherapy, radiation, and/or immunotherapy toxicities from 
recent chemotherapy would be known to the primary oncologist, and we would interpret AEs 
attributed to the study regimen with this knowledge in mind.  For example, if a patient received 
cytotoxic chemotherapy known to cause leukopenia prior to development of COVID-19, 
development of leukopenia following initiation of study therapy will not be considered treatment-
related, depending on the expected degree and duration of leukopenia following chemotherapy.
10.2.3 Toxicity Management
The toxicity of high dose interferon (20 million units/m²/d) has been established by Kirkwood et 
al in a number of trials. Most notable was the E1684 trial (47) where HDI (20 million units/m²/d) 
was administered daily for 5 days x 4weeks. In that trial (n=143), grade 3 toxicities were 67%, 
grade 4 toxicities were 9% (mainly constitutional and neurologic), and there were 2 treatment 
related mortalities (grade 5) due to hepatotoxicity. The proportion of Grade 3 and 4 toxicities in 
that trial were 48.2% for constitutional toxicities (defined as ‘worst grade of any constitutional 
toxicity, including fever, chills, flu-like symptoms, fatigue, malaise, and diaphoresis), and 66% for 
non-constitutional toxicities (23.8% for myelosuppression, 13.9% for hepatotoxicity, 28% for 
neurological toxicity).
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While the combination of HDI and rintatolimod may lead to synergistic additive side effects, we 
will monitor subject toxicities for unexpected increases in constitutional symptoms, 
myelosuppression, hepatotoxicity, and neurological toxicity. We will focus on monitoring for 
grade 3 or greater treatment related hematologic toxicities, and grade 4 or greater treatment related 
non-hematologic toxicities. A non-informative prior distribution of SAEs will be utilized to 
develop the safety stopping rules (details in Section 19). 
To our knowledge, the combination of HDI and rintatolimod has not been tested previously in 
patients with COVID-19. Therefore, we may observe adverse events that were not observed in 
trials of cancer and specific adverse events may be more severe in patients with COVID-19. One 
possibility is worsening of acute lung injury or systemic inflammatory response. Though the 
protocol does not specify specific treatments for such AEs, agents used for cytokine release 
syndrome, e.g. Tocilizumab, or JAK2, TNFα, IL-1 inhibitors, may be used, based on physician 
judgment.

Gastrointestinal Toxicity
o
Nausea and/or vomiting should be controlled with adequate antiemetic therapy. 
Prophylactic antiemetic therapy can be used at the discretion of the 
investigator/sub-investigator. Subjects are encouraged to take plenty of oral fluids.
o
Diarrhea should be managed with appropriate antidiarrheal therapy. Subjects 
should be encouraged to take plenty of oral fluids. If symptoms do not decrease to 
grade 1 or less with adequate antidiarrheal therapy, all protocol drugs should be 
held until resolved to < grade 1.

Pain
o
For fever or mild local pain, acetaminophen will be used at the discretion of the 
investigator/sub-investigator or designee.

Immunogenicity risk: Given the limited exposure to interferon alpha-2b (administered on 
Days 1 and 3), the risk of anti-drug-antibodies is very low. Hypersensitivity reaction can 
occur (described below). Our plan is to assess all AEs, including immunologic 
complications, and to provide appropriate management.

Hypersensitivity Reactions
Caution: Subjects who had a mild to moderate hypersensitivity reaction have been successfully 
re-challenged, but careful attention to prophylaxis and bedside monitoring of vital signs is 
recommended. Hypersensitivity reactions to interferon alpha-2b and/or rintatolimod will be 
managed as follows:
o
Mild symptoms (e.g., mild flushing, rash, pruritus): Complete infusion. Supervise 
at bedside. No treatment required.
o
Moderate symptoms (e.g., moderate rash, flushing, mild dyspnea, chest 
discomfort): Stop infusion. Give intravenous diphenhydramine 25 mg and 
intravenous dexamethasone 10 mg. Resume infusion after recovery of symptoms 
at a low rate, then, if no further symptoms, at full dose rate until infusion is 
complete. If symptoms recur, stop infusion. The subject should receive no 
additional interferon alpha-2b or rintatolimod. Record toxicity.
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o
Severe life-threatening symptoms (e.g., hypotension requiring pressor therapy, 
angioedema, respiratory distress requiring bronchodilation therapy, generalized 
urticaria): Stop infusion. Give intravenous diphenhydramine and dexamethasone as 
above. Add epinephrine or bronchodilators if indicated. If wheezing is present, that 
is not responsive to bronchodilators, epinephrine is recommended. Subject should 
be removed from further protocol therapy. Report as serious adverse event.
10.3 General Concomitant Medication and Supportive Care
There are no restrictions on general supportive care, which will be administered based on newest 
standards of COVID-19 care.
10.4 Duration of Study Treatment
A total of 2 daily doses over a 4-day day period of treatment is planned. 
The participant may stop treatment earlier than planned in the event that the following occurs:

Unacceptable toxicity

Progression to severe COVID-19 infection, defined by new or worse pulmonary infiltrates 
on Chest X-Ray or CT imaging plus one of the following: room air Sao2 ≤ 92%, room air 
PO2 <70 mm Hg, or PAO2-PaO2 ≥35 mm Hg, and/or requiring mechanical ventilation

Withdrawal of consent from study

Participant non-compliance with study requirements
11 PROCEDURES INVOLVED
Informed consent MUST be completed prior to receiving any study related procedures.
To limit exposure of COVID-19 to staff, patients will be provided with a surgical mask on entry 
to Roswell Park. The patient will be escorted to the site of drug administration using a dedicated 
elevator and immediately placed in a room with the door closed. All staff in contact with patients 
will don PPE. Infusions will be administered in a room with a closed door. History will be obtained 
by trained staff by phone in advance of study-related visits. Physical exam will be limited to vitals 
and auscultation of lungs. Following completion of treatment, the patient will be escorted to his/her 
vehicle by trained staff. A specific SOP will be developed to minimize contact between study 
subjects and staff and patients.
11.1 Screening
Within 3 days prior to treatment initiation and may be the same day as initiation of study treatment:

Recording of concomitant medications

Medical and Surgical History

Physical exam (lung auscultation), Vital Signs and weight; SaO2

ECOG Performance Status Assessment

Hematology

Chemistry

Arterial blood gas
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
Serum HCG pregnancy testing (within 3 days of Day 1 in women of child-bearing potential 
only)

EKG

Chest X-ray or CT imaging (NOTE: If a chest CT is ordered as standard of care prior to 
starting treatment, a chest x-ray is not required)

Nasopharyngeal swab

Evaluations Performed on each day of treatment [(Day 1and Day 3 (+ 1 day)]: Recording 
of concomitant medications

Recording of Adverse Events

Targeted physical exam
o
Auscultation of lungs
o
Vital Signs (temperature, blood pressure, pulse rate and respiratory rate, SaO2) will 
be performed pre- treatment and at 30 and 60 minutes post rintatolimod (± 5 
minutes)

ECOG Performance Status Assessment

Hematology

Chemistry

Blood (30cc) for correlative studies: On Day 1 treatment and Day 3 treatment, prior to 
treatment and within 1 hour after end of infusion treatment (two draws on each of these 
days). 

Nasopharyngeal swabs for viral quantification by quantitative PCR
11.2 Follow-up Visits on Days 7 (± 1 day) and 14 (± 3 days) 

Recording of concomitant medications

Recording of Adverse Events

Targeted physical exam
o
Lung auscultation and Vital Signs

ECOG Performance Status

Hematology

Chemistry

Blood (30cc) for correlative studies

Nasopharyngeal swabs for viral quantification by quantitative PCR
11.3 Correlative studies (blood samples)
Three (10 mL) green-top heparinized tubes and one red-top tube of blood will be collected via 
venipuncture for biomarker analysis at multiple time points (please refer to Appendix F: Study 
Calendar).
Tubes will be labeled with the participant’s initials, participant’s study number, clinical study 
number, time of collection and protocol day. Samples will be sent at room temperature to the 
attention of Laboratory Medicine – Protocol Clinical Research Support (pneumatic Station 19) 
where they will get accessioned for tracking. Once specimen receipt has been documented, the 
specimens will be sent at ambient temperature to pneumatic station 641 (located in CCC LOB-4th 
floor). The Kalinski Laboratory will be notified via telephone and e-mail (all contacts to be copied 
with each e-mail: see contact information below) prior to sample shipment and, the samples will 
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be held in CSPO until a response is received acknowledging that personnel are available to procure 
the samples from the pneumatic tube station. Samples will be processed and stored in the Kalinski 
Laboratory for future analysis.
Roswell Park Comprehensive Cancer Center
Kalinski Laboratory
CCC Bldg. 5th Floor, Rm. 502F
Attn: Study Number: I-659920
Elm & Carlton Streets
Buffalo, NY 14263
Tel: 716-845-1300 ext. 7353, ext.5175
Cell: 716-353-5384
melissa.grimm@roswellpark.org 
ronald.slomba@roswellpark.org 
pawel.kalinski@roswellpark.org 
Preliminary analyses will include the pre- and post-treatment cytokine levels and immune cell 
subsets, via RT-PCR, flow cytometry assays, or alternative laboratory methods. . Cytokine 
measurements will involve ELISAs using validated antibodies and will include generation of a 
standard curve with known quantities of the analyte and use of appropriate specificity controls, 
as recommended by the manufacturer. For cryopreserved samples, we will test for batch effects 
and repeat measurements on 10% of samples over a 3-month period to rule out loss of analyte 
detection by cryopreservation.
Below is a general description of processing of blood that might be modified per  Dr. Puzanov’s 
judgment. Plasma samples are to be separated from whole blood and processed at 4°C using a 
refrigerated centrifuge at approximately 3000 rpm for 10 minutes with the plasma (0.5 ml aliquots 
– as many as available) being aliquoted into cryovials per time-point. The samples will 
immediately be frozen at -70ºC or below until analyzed. PBMCs will then be obtained by Ficoll 
gradient as per the lab’s standard procedure. Two of the tubes will be pooled and the PBMCs will 
be frozen in Gibco Cell culture freezing media and will be stored in LN2 until time of analysis. 
The remaining tube of PBMCs will be frozen in RNA later and will be stored in -70 degrees or 
below until analyzed. Serum will be collected from red top tubes and 0.5 ml aliquots (as many as 
available) will be frozen at -70ºC. The screw cap polypropylene cryogenic tubes will be labeled 
with the clinical study number, participant’s MR number, participant’s study number, protocol 
time point, protocol day, date and time of draw.
As a component of exploratory aims, blood will be analyzed to assess pre- and post- treatment 
chemokine and immune cell subsets, via ELISA, RT-PCR, and flow cytometry assays to obtain 
insights into the synergy between rintatolimod and individual concentrations of IFNα.
In the induction of known mediators of anti-coronavirus responses and factors of intracellular 
immunity against RNA viral infections in general, we may also evaluate the levels of expression 
of myxovirus resistance gene (MxA), protein Kinase R (PKR); oligoadenylate synthetase-
2  (OAS2); RNAse-L; IFN-stimulated gene-15 (ISG15); IFN-induced proteins with 
tetratricopeptide repeats (IFIT1) and IFN-inducible transmembrane protein 3 (IFITM3), as well 
TLR3, RIG-I, MDA5, IRF3, IRF7,  in nasopharyngeal swabs material and blood cells of patients 
on all tiers of treatment, using RT-PCR and, potentially, intracellular protein expression (flow 
cytometry and image stream).
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Note: All investigator or analyzing research laboratories housing research samples need to 
maintain current Temperature Logs and study-specific Sample Tracking and Shipping Logs. 
The Principal Investigator/Laboratory Manager must ensure that the stated lab(s) have a process 
in place to document the receipt/processing/storage/shipping of study-related samples/specimens. 
This is required for both observational and interventional clinical studies collecting clinical 
samples.
11.4 Nasopharyngeal swabs
Nasopharyngeal swabs will be taken for viral quantification by quantitative PCR on:

Screening: If a potential study participant has already been identified as COVID-19-
positive from an outside testing facility, as per current institutional protocol, a 
nasopharyngeal swab will be obtained for quantitative PCR. Study drug therapy will be 
initiated within 3 days of screening, including on the same day of screening.

Each day of treatment (Day 1 and Day 3 (+ 1 day). 

Follow-up Visits on Days 7 and 14
Testing for SARS-CoV-2 will be done upfront and residual samples will be tested for baseline 
viral load. If there’s a problem in supplies, repeat sampling at the indicated time points may not be 
feasible and will not be considered a protocol deviation.
All samples will be transported to the Molecular Diagnostics facility (Attn: Dr. Jan Nowak) The 
screening samples will be tested for SARS-CoV-2 as per current institutional protocol; The 
residual sample left over from clinical testing, as well as from subsequent testing, will be processed 
by the Molecular Diagnostics facility and the resulting research analytes will be transported to the 
Kalinski laboratory for storage and analysis (CCC Bldg. 5th Floor, Rm. 502F).
Any remaining RNA may be also analyzed for the markers of antiviral immunity described in the 
blood correlative studies.
12 WITHDRAWAL OF SUBJECTS
12.1 Treatment Discontinuation
Upon treatment discontinuation all end of treatment evaluations and tests will be conducted. All 
participants who discontinue due to an AE must be followed until the event resolves or stabilizes. 
Appropriate medical care should be provided until signs and symptoms have abated, stabilized, or 
until abnormal laboratory findings have returned to acceptable or pre-study limits. The final status 
of the AE will be reported in the participant’s medical records and the appropriate eCRF.
Reasons for treatment discontinuation should be classified as follows:

Death

Toxicity: treatment related or unrelated

Participant non-compliance with study requirements

Investigator judgment
o
The Investigator may discontinue a participant if, in his/her judgment, it is in the 
best interest of the participant to do so.

Participant voluntary withdrawal
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13 RISKS TO SUBJECTS
13.1 Interferon Alpha-2b
Interferon alpha-2b may cause fever, chills and flu-like symptoms; loss of appetite; nausea; 
vomiting, diarrhea and abdominal pain; fatigue; lowered white blood count may increase risk of 
infection; lowered platelets may lead to an increase in bruising or bleeding; hair loss. Other risks 
which may be common in cases of prolonged administration of interferon alpha-2b include 
drowsiness; temporary confusion; anxiety, amnesia, irritability, confusion, delusions and 
depression which can be severe; numbness and/or tingling in the hands and/or feet, skin rashes and 
inflammation of the pancreas. Inflammation of the pancreas is swelling or irritation of the pancreas 
which may result in tenderness or pain in the stomach and/or back. When the pancreas is inflamed, 
the body is not able to absorb all the nutrients it needs.
Additional information on risk is included in the Investigator’s Brochure or package insert.
13.2 Rintatolimod
Clinical experience with rintatolimod totals over 800 patients with more than 400 patients 
receiving rintatolimod for at least six (6) months, greater than 200 patients for one (1) year, over 
50 patients up to two (2) years, and with 20 or more patients over two (2) years at doses as high as 
1200 mg i. v. twice weekly. No evidence of dose-limiting organ toxicity, including hematologic, 
liver, or renal toxicity, has been observed.
Adverse events related to infusion such as mild flu-like symptoms, transient headache, fever, 
myalgia, arthralgia, and fatigue/malaise which were seen, usually occur during the initial weeks of 
treatment and tend to subside on repeated administration. These side events were seen in Chronic 
Fatigue Syndrome patients, cancer patients, chronic hepatitis B infected patients and individuals 
infected with HIV at doses of 200 and 400 mg and higher. Patients that experience these minor 
side effects can continue on rintatolimod and as noted, these signs and symptoms typically subside 
after several weeks of continued treatment. Specific symptoms of note include a flushing reaction, 
characterized by at least one occurrence of erythema of the face, neck and chest, which has been 
observed in approximately 10% of patients treated in various studies. Usually the flushing is both 
mild and transient and disappears with repeated dosing. Occasionally, it can be accompanied by a 
tightness of the chest, tachycardia, anxiety, shortness of breath, subjective reports of "feeling hot", 
diaphoresis and nausea. The reaction is usually infusion-rate dependent and can generally be 
controlled by slowing the infusion rate. An antihistamine (diphenhydramine hydrochloride) can 
be helpful in controlling and reducing the response in the occasional patient for whom the symptom 
persists. Other less frequently occurring adverse effects include nausea, diarrhea, itching, urticaria, 
bronchospasm, transient hypotension, photophobia, rash, bradycardia, and transient visual 
disturbances. A severe unexpected local reaction to extravasation of rintatolimod (Ampligen®, 
Poly I:Poly C12U) at the infusion site in the dorsum of the left hand was reported in a Chronic 
Fatigue Syndrome patient with chilblains. Several patients experienced liver enzyme level 
elevations while receiving rintatolimod associated with chronic dosing over many weeks.
Rintatolimod has been dosed in combination with alpha interferon in investigator-initiated studies 
under investigator IND applications during the period between December 1985 and April 1994. A 
total of 24 patients received combination treatments. Clinical conditions included renal cell 
carcinoma, chronic myelogenous leukemia, melanoma, and ovarian cancer. Rintatolimod was 
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given as an IV infusion at a dose of 300 mg twice weekly. The starting dose was sometimes as low 
as 1-10 mg. The interferons were administered at a dose of 3 million Units daily, with some doses 
of 0.75 million units at the low side and up to 6 million units at the higher side.
The therapy with rintatolimod in combination with alpha interferon was generally well tolerated 
without evidence of dose-limiting or cumulative toxicities. The most frequent adverse reactions 
were considered minor in severity and duration. Most of them were flu-like symptoms such as 
chills, cold feeling, fatigue, decreased appetite, fever, muscular aches. Also, shortness of breath 
has been seen as well as hypotension, nausea, anemia, dyspnea, numbness, itching and blurred 
vision. These adverse events were judged possibly related to the condition of the patients, but also 
possibly related to the administration of rintatolimod or interferon. In some cases, worsening of 
the patient’s condition has been seen due to tumor progression, but in general favorable clinical 
patterns were observed in these patients with advanced disease. Such combinations were not 
intended to be immune modulating and were evaluated in the context of clinical cancer care.
For more risk information, reference the Investigator’s Brochure or package insert.
14 POTENTIAL BENEFITS TO SUBJECTS
Patients enrolled in this study may demonstrate improved virus control, manifested by reduced 
symptoms and improved oxygenation or lack of progression of infection to pneumonia and ARDS. 
15 DATA AND SPECIMEN BANKING
All samples for correlative analysis will be sent to the Kalinski laboratory for processing, storage, 
and analysis. Samples will be used for planned study assays as well as for future studies that 
include, but are not limited to, immune responses to viral infections and to immunotherapy.
Any clinical data that is associated with the samples, will be stored on a secure server in the 
Department of Medicine, will be accessible only by the PI, Co-Investigators and PI designated 
data manager and, will be password protected. All computer entry and networking programs will 
be done using PIDs only. Any clinical data and/or specimens for future research will require 
verification of an IRB-approved protocol and will be de-identified before being released to 
investigators within and outside of Roswell Park.
Note: All investigator or analyzing research laboratories housing research samples need to 
maintain current Temperature Logs and study-specific Sample Tracking and Shipping Logs. The 
Principal Investigator/Laboratory Manager must ensure that the stated lab(s) have a process in 
place to document the receipt/processing/storage/shipping of study-related samples/specimens. 
This is required for both observational and interventional clinical studies collecting clinical 
samples.
16 MEASUREMENT OF EFFECT
16.1 Primary Endpoints

The primary safety endpoints are frequency of grade 3 or 4 AEs considered to be possibly, 
probably, or definitely related to the treatment regimen. 

Evaluate kinetics of viral load in nasopharyngeal swabs based on quantitative PCR in the 
course of treatment and Days 7, and 14.
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16.2 Secondary Endpoints

Clinical efficacy will be assessed by the frequency of these complications: (i) progression 
of infection requiring hospitalization; (ii) respiratory failure requiring mechanical 
ventilation (primary efficacy endpoint); and (iii) death within 30 days. If present, acute 
respiratory distress syndrome (ARDS) will be graded by Berlin criteria (42).

Kinetics of viral load in the peripheral blood and nasopharyngeal swab based on 
quantitative PCR in the course of treatment and Days 7 and 14.

Kinetics of changes of the immune subsets and circulating inflammatory mediators 
(including CRP, cytokines, chemokines, interferons) in peripheral blood in the course of 
treatment and at Days 7 and 14.

Induction of known mediators of antiviral immunity that include (myxovirus resistance 
gene, MxA; protein Kinase R (PKR); oligoadenylate synthetase-2  (OAS2); RNAse-L,  
IFN-stimulated gene-15 (ISG15);  IFN-induced proteins with tetratricopeptide repeats 
(IFIT1) and IFN-inducible transmembrane protein 3 (IFITM3), TLR3, RIG-I, MDA5, 
IRF3, IRF7,  in nasopharyngeal swabs material and blood cells of patients on all tiers of 
treatment. Expression of ACE2 (receptor for SARS-Cov-2 entry) and potentially other 
genes involved in SARS-CoV-2 infection will be tested in nasopharyngeal samples.
17 SAFETY EVALUATION
17.1 Adverse Events
An adverse event or adverse experience (AE) is any untoward medical occurrence associated with 
the use of a drug in humans, whether or not considered drug related. Therefore, an AE can be ANY 
unfavorable and unintended sign (including an abnormal laboratory finding), symptom, or disease 
temporally associated with the use of a medicinal (investigational) product, whether or not 
considered related to the medicinal (investigational) product (attribution of ‘unrelated’, ‘unlikely’, 
‘possible’, ‘probable’, or ‘definite’).
An AE is considered “unexpected” if it is not listed in the investigator brochure or is not listed at 
the specificity or severity that has been observed; or if an investigator brochure is not required or 
available, is not consistent with the risk information described in the general investigational plan 
in other study-related documents.
17.2 Diagnosis Versus Signs and Symptoms
If known, a diagnosis should be recorded on the CRF rather than individual signs and symptoms 
(e.g., record only liver failure or hepatitis rather than jaundice, asterixis, and elevated 
transaminases). However, if a constellation of signs and/or symptoms cannot be clinically 
characterized as a single diagnosis or syndrome at the time of reporting, each individual event 
should be recorded as an AE or SAE on the CRF. If a diagnosis is subsequently established, it 
should be reported as follow-up information.
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17.3 Adverse Events Occurring Secondary to Other Events
In general, AEs occurring secondary to other events (e.g., cascade events or clinical sequelae) 
should be identified by their primary cause. For example, if severe diarrhea is known to have 
resulted in dehydration, it is sufficient to record only diarrhea as an AE or SAE on the CRF.
However, clinically significant AEs occurring secondary to an initiating event that are separated 
in time should be recorded as independent events on the CRF. For example, if a severe 
gastrointestinal hemorrhage leads to renal failure, both events should be recorded separately on 
the CRF.
17.4 Abnormal Laboratory Values
Only clinically significant laboratory abnormalities that require active management will be 
recorded as AEs or SAEs on the CRF (e.g., abnormalities that require study drug dose 
modification, discontinuation of study treatment, more frequent follow-up assessments, further 
diagnostic investigation, etc.).
If the clinically significant laboratory abnormality is a sign of a disease or syndrome (e.g., alkaline 
phosphatase and bilirubin 5 x the upper limit of normal associated with cholecystitis), only the 
diagnosis (e.g., cholecystitis) needs to be recorded on the Adverse Event CRF.
If the clinically significant laboratory abnormality is not a sign of a disease or syndrome, the 
abnormality itself should be recorded as an AE or SAE on the CRF. If the laboratory abnormality 
can be characterized by a precise clinical term, the clinical term should be recorded as the AE or 
SAE. For example, an elevated blood potassium level of 7 mEq/L should be recorded as 
“hyperkalemia”.
Observations of the same clinically significant laboratory abnormality from visit to visit should 
not be repeatedly recorded as AEs or SAEs on the CRF, unless their severity, seriousness, or 
etiology changes.
17.5 Preexisting Medical Conditions (Baseline Conditions)
A preexisting medical condition should be recorded as an AE or SAE only if the frequency, 
severity, or character of the condition worsens during the study. When recording such events on 
an Adverse Event CRF, it is important to convey the concept that the preexisting condition has 
changed by including applicable descriptors (e.g., “more frequent headaches”).
17.6  Grading and Reporting Adverse Events
17.6.1 Grading and Relationship to Drug
The descriptions and grading scales found in the CTEP Version 5.0 of the NCI Common 
Terminology Criteria for Adverse Events (CTCAE) will be utilized for AE reporting. CTEP 
Version 5.0 of the CTCAE is identified and located at:
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm.
AEs not covered by specific terminology listed should be reported with common medical 
terminology and documented according to the grading scales provided in the CTCAE Version 5.0.
The relationship of event to study drug will be documented by the Investigator as follows:
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Unrelated: The event is clearly related to other factors such as the participant’s clinical state, other 
therapeutic interventions or concomitant drugs administered to the participant.
Unlikely: The event is doubtfully related to investigational agent(s). The event was most likely 
related to other factors such as the participant’s clinical state, other therapeutic interventions, or 
concomitant drugs.
Possible: The event follows a reasonable temporal sequence from the time of drug administration 
but could have been produced by other factors such as the participant’s clinical state, other 
therapeutic interventions, or concomitant drugs.
Probable: The event follows a reasonable temporal sequence from the time of drug administration 
and follows a known response pattern to the study drug. The event cannot be reasonably explained 
by other factors such as the participant’s clinical state, therapeutic interventions, or concomitant 
drugs.
Definite: The event follows a reasonable temporal sequence from the time of drug administration, 
follows a known response pattern to the study drug, cannot be reasonably explained by other 
factors such as the participant’s condition, therapeutic interventions or concomitant drugs; AND 
occurs immediately following study drug administration, improves upon stopping the drug, or 
reappears on re-exposure.
17.6.2 Reporting Adverse Events
Routine AEs occurring between the start date of intervention until 30 days after the last 
intervention, or until the event has resolved, the study participant is lost to follow-up, the start of 
a new treatment, or until the study investigator assesses the event(s) as stable or irreversible, will 
be reported. New information will be reported after it is received.
Guidelines for Routine Adverse Event Reporting for Phase 1 Studies (Regardless of 
Expectedness)
Attribution
Grade 1
Grade 2
Grade 3
Grade 4
Unrelated
X
X
X
X
Unlikely
X
X
X
X
Possible
X
X
X
X
Probable
X
X
X
X
Definite
X
X
X
X
Guidelines for Routine Adverse Event Reporting for Pilot, Phase 2, and Phase 3 Studies 
(Regardless of Expectedness)
Attribution
Grade 1
Grade 2
Grade 3
Grade 4
Unrelated
X
X
Unlikely
X
X
Possible
X
X
X
X
Probable
X
X
X
X
Definite
X
X
X
X
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17.7 Serious Adverse Events
A serious adverse event (SAE) is any adverse event (experience) that in the opinion of either the 
investigator or sponsor results in ANY of the following:

Death.

A life-threatening adverse event (experience). Any AE that places a participant or 
participants, in the view of the Investigator or sponsor, at immediate risk of death from the 
reaction as it occurred. It does NOT include an AE that, had it occurred in a more severe 
form, might have caused death.

Inpatient hospitalization or prolongation of existing hospitalization (for > 24 hours).

A persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions.

A congenital anomaly or birth defect.

Important Medical Event (IME) that, based upon medical judgment, may jeopardize the 
participant and may require medical or surgical intervention to prevent one of the outcomes 
listed above.
17.7.1 Reporting Serious Adverse Events
All new SAEs occurring from the date the participant signs the study consent until 30 days after 
the last intervention or a new treatment is started, whichever comes first, will be reported. The 
Roswell Park SAE Source Form is to be completed with all available information, including a brief 
narrative describing the SAE and any other relevant information.
SAEs occurring after the 30 day follow-up period that the investigator determines to be possibly, 
probably, or definitely related to the study intervention should be reported.
SAEs identified as an Unanticipated Problem by the Investigator must be reported. Please refer to 
Section 17.10 for details on reporting Unanticipated Problems.
17.8 Follow-Up for Serious Adverse Events
All related SAEs should be followed to their resolution, until the study participant is lost to 
follow-up, the start of a new treatment, or until the study investigator assesses the event(s) as stable 
or irreversible. New information will be reported when it is received.
17.9 Unanticipated Problems
An Unanticipated Problem (UP) is any incident, experience, or outcome that meets all of the 
following criteria:

Unexpected (in terms of nature, severity, or frequency) given:
o
The research procedures that are described in the study-related documents, 
including study deviations, as well as issues related to compromise of participant 
privacy or confidentiality of data.
o
The characteristics of the participant population being studied.

Related or possibly related to participation in the research (possibly related means there is 
a reasonable possibility that the incident, experience, or outcome may have been caused by 
the procedures involved in the research).
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
Suggests that the research places participants or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized and if in relation to an AE is also deemed Serious per Section 17.7.
17.9.1 Reporting Unanticipated Problems
The Reportable New Information (RNI) Form will be submitted to the CRS Quality Assurance 
(QA) Office within 1 business day of becoming aware of the Unanticipated Problem. After review, 
CRS QA Office will submit the RNI to the IRB.
When becoming aware of new information about an Unanticipated Problem, submit the updated 
information to CRS QA Office with an updated Reportable New Information Form. The site 
Investigator or designated research personnel will report all unanticipated problems to the IRB in 
accordance with their local institutional guidelines.
17.10 FDA Reporting
When Roswell Park is the IND holder the following describes the FDA reporting requirements by 
timeline for AEs and new safety findings that meet the criteria outlined below:
Within 7 Calendar Days
Any adverse event that meets ALL the following criteria:

Related or possibly related to the use of the study drug.

Unexpected; and

Fatal or life-threatening.
Within 15 Calendar Days
Any adverse event that meets ALL the following criteria:

Related or possibly related to the use of the study drug.

Unexpected; and

Serious but not fatal or life-threatening.
Or, meets ANY of the following criteria:

A previous adverse event that is not initially deemed reportable but is later found to fit the 
criteria for reporting (report within 15 days from when event was deemed reportable).

Any findings from other studies, including epidemiological studies, pooled analysis of 
multiple studies, or other clinical studies conducted with the study drug that suggest a 
significant risk in humans exposed to the drug.

Any findings from animal or in vitro testing that suggest a significant risk for human 
participants including reports of mutagenicity, teratogenicity, or carcinogenicity or reports 
of significant organ toxicity at or near the expected human exposure.

Any clinically important increase in the rate of occurrence of a serious, related or possibly 
related adverse event over that listed in the protocol or investigator brochure.
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Sponsors are also required to identify in IND safety reports, all previous reports concerning similar 
adverse events and to analyze the significance of the current event in the light of the previous 
reports.
Reporting Process
The principal investigator or designee will complete and submit a FDA Form 3500A MedWatch 
for any event that meets the above criteria. Forms will be submitted to the CRS QA Office via 
email to CRSQA@RoswellPark.org.
18 DATA MANAGEMENT AND CONFIDENTIALITY
18.1 Data Collection
Full build studies are managed by Roswell Park CRS Data Management for analysis by Roswell 
Park Biostatisticians. All electronic case report form (eCRF) data are captured for these studies.
Data management activities are performed using a CTMS system that enables the collection, 
cleaning and viewing of clinical trial data. CRS data management designs the study-specific 
database and facilitates development by the Information Technology team. Once the database 
design is approved by the Investigator, Statistician, and Clinical Research Coordinator, the 
database is put into production and data entry can begin. Data can be entered and changed only by 
those with the rights to do so into the eCRFs.
18.2 Maintenance of Study Documents
Essential documents will be retained per Roswell Park’s policy for 6 years from the study 
termination date. These documents could be retained for a longer period, however, if required by 
the applicable local regulatory requirements or by an agreement with Roswell Park.
18.3 Revisions to the Protocol
Roswell Park may make such changes to the protocol as it deems necessary for safety reasons or 
as may be required by the U.S. FDA or other regulatory agencies. Revisions will be submitted to 
the IRB/ERC for written approval before implementation.
18.4 Termination of the Study
It is agreed that, for reasonable cause, either the Roswell Park Investigators or the Sponsor, may 
terminate this study, provided a written notice is submitted within the time period provided for in 
the Clinical Trial Agreement. In addition, Roswell Park may terminate the study at any time upon 
immediate notice if it believes termination is necessary for the safety of participants enrolled in 
the study.
19 STATISTICAL PLAN
19.1 Study Design/Endpoints
This is prospective Phase I/IIa trial basket trial of i.v. rintatolimod administered with or without 
i.v. IFNα (INTRON® A) in two cohorts of patients:  Main Cohort and Expansion Cohort. Only 
Main Cohort subjects will participate in the Phase I  3+3 portion of this study. The Expansion 
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Cohort will consist of rintatolimod without IFNα. Neither cohort is randomized. The kinetics of 
nasopharyngeal viral load will be assessed based on baseline values and on treatment and EOT 
values. In the Expansion Cohort, the kinetics of nasopharyngeal viral load will be compared with 
historical controls consisting of Roswell Park patients with COVID-19 of varying severity. All 
patients will receive best available standard of care in addition to study drug regimens. 
Phase I. The Phase I component of the protocol consists of a 3+3 design with four dose levels of 
IFNα (0, 5 MU/M2, 10 MU/M2  and 20 MU/M2  ) and fixed dose rintatolimod (200 mg per day) 
administered on Days 1 and 3.  The sample size for the Phase I trial may range from n=3 to 24.
Primary Endpoint

The primary safety endpoint is the frequency of grade 3 or 4 AEs considered to be 
possibly, probably, or definitely related to the treatment regimen.
Phase IIa. The Phase IIa component of the trial is designed to be a single arm trial of IFNα (at 
MTD, if reached, otherwise at 20 MU/M2) + rintatolimod along with the best available standard 
of care. Using each patient’s baseline nasopharyngeal viral load, we will evaluate to what extent 
the study regimen reduces viral load relative historical institutional benchmarks. The sample size 
will be n=10.
Primary Endpoint

Evaluate kinetics of viral load expressed as Ct values in nasopharyngeal swab samples over 
time measured at baseline (day 0), Days 3 (last day of treatment), 7, and 14 (40). The 
primary endpoint will be the area under the Ct curve (AUC).
The primary Phase IIa safety endpoint is the frequency of grade 3 or 4 AEs considered to be 
probably or definitely related to the treatment regimen.
Secondary Endpoints

Clinical efficacy will be assessed by the frequency of these complications: (i) progression 
of infection requiring hospitalization; (ii) respiratory failure requiring mechanical 
ventilation; (iii) death within 30 days and (iv) Time to a Ct value of non-detectable.
Correlative Endpoints

Determine the kinetics of changes of the immune subsets and circulating inflammatory 
mediators (including CRP, cytokines, chemokines, interferons) in peripheral blood in the 
course of treatment at baseline (Day 0), Days 3, 7, and 14. Plasma and PBMCs will be 
cryopreserved for future exploratory studies, including the assessment of intracellular 
signaling and TCR and BCR repertoires as correlates of response to the study regimen.

Induction of known mediators of antiviral immunity that include (myxovirus resistance 
gene, MxA; protein Kinase R (PKR); oligoadenylate synthetase-2  (OAS2); RNAse-L,  
IFN-stimulated gene-15 (ISG15);  IFN-induced proteins with tetratricopeptide repeats 
(IFIT1) and IFN-inducible transmembrane protein 3 (IFITM3), TLR3, RIG-I, MDA5, 
IRF3, IRF7, in nasopharyngeal swabs material and blood cells of patients on all tiers of 
treatment. Expression of ACE2 (receptor for SARS-Cov-2 entry) and potentially other 
genes involved in SARS-CoV-2 infection will be tested in nasopharyngeal samples.
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All subjects meeting the eligibility criteria who signed a consent form and  receive rintatolimod 
control will be considered evaluable for efficacy analysis. Safety analysis will be performed on all 
patients who have signed a consent form and received at least one dose of study medication.
The primary efficacy analysis of the Phase IIa component of the study will be on the intention to 
treat (ITT) population, which will include all patients who have signed a consent form and 
undergone randomization, according to the treatment group to which they were allocated.
All data collected will be summarized and presented. Continuous variables will be described as 
the mean, median, standard deviation, and range of the observations. Categorical data will be 
described with contingency tables including frequency and percentage. Individual patient listings 
will be generated and presented.
Statistical descriptions and analyses will be carried out using SAS statistical analysis software 
version 9.4 or higher (SAS Institute, Inc., Cary, North Carolina, USA).
19.2 Proposed Data Analysis
19.2.1 Safety
SAFETY RUN-IN DESIGN
We will use a 3+3 dose escalation scheme in order to determine the safe dose to utilize in our 
efficacy analysis. The 3+3 design will consider four dose levels of IFNα (0, 5 MU/M2, 10 MU/M2 
and 20 MU/M2) and fixed dose rintatolimod (200 mg per day) administered on Days 1 and 3.
For each cohort, the given dose level will start with three patients, with three patients added to any 
dose level if we observe 1 out of 3 DLTs. The patients in each cohort will be staggered, so that the 
first patient in each cohort will have received treatment on Days 1 and 3 before the following 
patient(s) are enrolled in that cohort. If 2 out of 3 to 6 subjects have a DLT we will not proceed to 
the next dose level.
19.2.2 Efficacy
In order to compare the AUC between rintatolimod versus historical institutional benchmarks we 
will use the highly efficient approach proposed by Chandrasekhar et al (48). Ct values measured 
at baseline (Day 0), Days 3, 7 and 14 will be used to measure the AUC using a standard 
trapezoidal rule where the AUC for the ith subject in group k (k=1 for rintatolimod and k=2 for 
control) as 𝐴𝑈𝐶𝑗𝑘= ∑𝑚
𝑖=1 𝑐𝑗 𝑌𝑖𝑗𝑘 where 𝑡1 = 0,𝑡2 = 3,  𝑡3 = 7, 𝑡4 = 14, 𝑡5 = 30,, 𝑚= 5, the 𝑌𝑖𝑗𝑘
’s denote the corresponding Ct values per subject at the respective time points and
𝑐𝑗=
𝑡𝑗+1 ― 𝑡𝑗
2
, 𝑗= 1
𝑡𝑗― 𝑡𝑗―1
2
, 𝑗= 𝑚
𝑡𝑗+1 ― 𝑡𝑗―1
2
, 𝑜𝑡ℎ𝑒𝑟𝑤𝑖𝑠𝑒
If a Ct value is >40 it will be considered right censored for the purpose of this analysis, which 
corresponds to a non-detectable level. We can re-express this formulation using a linear mixed 
model with details given exactly as in Chandrasekhar et al. (48).  A large AUC value for a 
respective subject is a measure of treatment efficacy as compared to a subject with a lower AUC 
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value. The primary test of efficacy will be H0: AUC=130 (log2 copies) x day versus H1: 
AUC>130 (log2 copies) x day.
The secondary clinical efficacy endpoints of (i) progression of infection requiring hospitalization 
in patients who were not inpatients at study entry; (ii) respiratory failure requiring mechanical 
ventilation; and (iii) death within 30 days will be compared using Fisher’s exact test. Time to a Ct 
value of non-detectable will be examined using an exact discrete time-to-event Cox model.  The 
analysis of correlative endpoints will be primarily descriptive and graphical in nature.
The primary test of efficacy will be one-sided and tested at level alpha=0.05. All additional tests 
will be two-sided and tested at level alpha=0.05.
19.3 Sample Size Justification
The sample size was based on preliminary data gathered at Roswell Park on patients and 
employees.  The values were similar between the two study groups. The Ct values collected at 
baseline for n=20 patients and employees had an average of 24.1 log2 copies with a standard 
deviation of 6.5.  The standard deviations after baseline was approximately 4.5 as the Ct values 
increased.  The preliminary estimate for the AUC based on our pilot data is 130 (log2 copies) x 
day, which is the basis for our null hypothesis.  The between time point correlation was estimated 
to be approximately rho=0.3.  Given n=10 subjects we will be able to detect and increase in the 
AUC to an average of 144  (log2 copies) x day (efficacy) or greater than our assumed institutional 
benchmark of 130 (log2 copies) x day at power=0.80 and alpha=0.05(one-sided). We simplified 
this calculation assuming no censoring, which will be accounted for in the final analysis and 
assumed a compound symmetry covariance structure with the correlation estimate described 
above.  
19.4 Randomization
This is a non-randomized trial. 
19.5 Study Duration and Compliance
All study drug administration and compliance data will be summarized.
19.6 Prior and Concomitant Medication
All relevant prior medication and all concomitant medications will be summarized by frequencies 
and percentages. All medications will be coded using the  MeDRA drug dictionary.
19.7 Safety Review and Study Suspension
The Roswell Park Data Safety and Monitoring Committee (DSMC) will assess the progress of the 
study, the safety data, and critical efficacy endpoints. The DSMC will review the study weekly 
and will make recommendations that include but not limited to; (a) continuation of the study, (b) 
modifications to the design (c) suspension of or, (d)  termination of the study.
Additionally, should the study be suspended due to safety; members of the research team, study 
statistician, and DSMC will meet to discuss safety concerns and possible adjustments to the study 
or study closure.
Therapy will be discontinued for any Grade 4 treatment-related toxicity.
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19.8 Procedure for Accounting for Missing, Unused and Spurious Data
Missing data will be indicated in the listings but excluded from all descriptive analyses. All data 
will be listed, including otherwise unused data. Spurious data will be identified as such, wherever 
possible.
20 PROVISIONS TO MONITOR THE DATA TO ENSURE THE SAFETY OF 
SUBJECTS
All Roswell Park Phase I studies are reviewed at the scheduled Early Phase Clinical Trial 
Committee meetings and the minutes are forwarded to the IRB for review.
In addition, the Roswell Park Data Safety Monitoring Committee will assess the progress of the 
study, the safety data, and critical efficacy endpoints (Phase I studies will be reviewed quarterly; 
Phase II, III and pilot investigator-initiated studies will be reviewed semi-annually). The DSMC 
will review the study and will make recommendations that include but not limited to; (a) 
continuation of the study, (b) modifications to the design (c) suspension of or, (d) termination of 
the study.
Additionally, should the study be suspended due to safety; members of the research team, study 
statistician, and DSMC will meet to discuss safety concerns and possible adjustments to the study 
or study closure.
21 VULNERABLE POPULATIONS
The following vulnerable populations will be excluded:

Chronically cognitively impaired adults (e.g. those with dementia) with impaired decision-
making capacity

Individuals who are not yet adults (infants, children, teenagers)

Pregnant women

Prisoners
22 COMMUNITY-BASED PARTICIPATORY RESEARCH
Not applicable.
23 SHARING OF RESULTS WITH SUBJECTS
Individual response data is shared with the participant as a part of their clinical care.
24 SETTING
All evaluations and procedures will be conducted on an outpatient basis at Roswell Park 
Comprehensive Cancer Center.
25 PROVISIONS TO PROTECT THE PRIVACY INTERESTS OF SUBJECTS
Any data, specimens, forms, reports, video recordings, and other records that leave the site will be 
identified only by a participant identification number (Participant ID, PID) to maintain 
confidentiality. All records will be kept in a limited access environment. All computer entry and 
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networking programs will be done using PIDs only. Information will not be released without 
written authorization of the participant.
26 RESOURCES AVAILABLE
Not applicable.
27 PRIOR APPROVALS
Not applicable.
28 COMPENSATION FOR RESEARCH-RELATED INJURY
If the subject believes they have been injured as a direct result of their participation in this research 
study, they will be advised to notify the Roswell Park Patient Advocate at (716) 845-1365 or the 
Study Doctor at (716) 845-5721.
Medical diagnosis and treatment for the injury will be offered, and a determination will be made 
regarding appropriate billing for the diagnosis and treatment of the injury. A financial counselor 
(716-845-3161) will be able to provide an explanation of coverage and to answer questions the 
subject may have regarding study related billing.
The subject is not prevented from seeking to collect compensation for injury related to malpractice, 
fault, or blame on the part of those involved in the research.
29 ECONOMIC BURDEN TO SUBJECTS
The participants will not be subject to any economic burden.
30 CONSENT PROCESS
The Roswell Park SOP: Informed Consent Process for Research (HRP-090) will be followed.
This study will not be initiated until the protocol and informed consent document(s) have been 
reviewed and approved by a properly constituted Institutional Review Board (IRB) or Independent 
Ethics Committee (IEC). Each participant (or legal guardian) shall read, understand, and sign an 
instrument of informed consent prior to performance of any study-specific procedure. It is the 
responsibility of the investigator to ensure that the participant is made aware of the investigational 
nature of the treatment and that informed consent is given.
The Investigator is responsible for the retention of the participant log and participant records; 
although personal information may be reviewed by authorized persons, that information will be 
treated as strictly confidential and will not be made publicly available. The investigator is also 
responsible for obtaining participant authorization to access medical records and other applicable 
study specific information according to Health Insurance Portability and Accountability Act 
regulations (where applicable).
This study will be conducted in compliance with all applicable laws and regulations of the state 
and/or country and institution where the participant is treated. The clinical trial should be 
conducted in accordance with the ethical principles embodied in the Belmont Report: Ethical 
Principles and Guidelines for the Protection of Human Subjects of Research, consistent with good 
clinical practice and the applicable regulatory requirements and according to the guidelines in this 
protocol, including attached appendices.
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31 PROCESS TO DOCUMENT CONSENT IN WRITING
The Roswell Park “SOP: Written Documentation of Consent (HRP-091)” will be followed.
The Investigator (or IRB specified designee) is responsible for obtaining written consent from each 
participant or the participant's legally authorized representative in accordance with GCP guidelines 
using the approved informed consent form, before any study specific procedures (including 
screening procedures) are performed. The informed consent form acknowledges all information 
that must be given to the participant according to applicable GCP guidelines, including the purpose 
and nature of the study, the expected efficacy and possible side effects of the treatment(s), and 
specifying that refusal to participate will not influence further options for therapy. Any additional 
information that is applicable to the study must also be included. Additional national or 
institutionally mandated requirements for informed consent must also be adhered to. The 
participant should also be made aware that by signing the consent form, processing of sensitive 
clinical trial data and transfer to other countries for further processing is allowed.
The Investigator or designee shall provide a copy of the signed consent form to the participant and 
the signed original shall be maintained in the Investigator File. A copy of the signed consent form 
must be filed in the participant file. At any stage, the participant may withdraw from the study and 
such a decision will not affect any further treatment options.
32 DRUGS OR DEVICES
Roswell Park will hold the IND for this study.
32.1 Interferon Alpha-2b
Interferon alpha-2b is approved around the world for the treatment of chronic hepatitis C, chronic 
hepatitis B, hairy cell leukemia, chronic myelogenous leukemia, multiple myeloma, follicular 
lymphoma, carcinoid tumor, and malignant melanoma. Interferon alpha-2b has many drug 
classifications 
including 
anti-infective, 
anti-neoplastic, 
antiproliferative, 
antiviral 
and 
immunological agent.
32.1.1 Drug Shipment
Interferon alpha-2b will be provided by Roswell Park and will be ordered from the manufacturer 
through IDS.
The date of receipt and the amount of drug received will be documented. Drug shipment records 
will be retained by the investigational pharmacist or designee.
32.1.2 Preparing and Dispensing
The lyophilized product is reconstituted as directed by the manufacturer. Investigational Drug 
Service Pharmacy (IDS) will prepare and dispense.
For IV injection, it is recommended that interferon alpha-2b be administered as a 100,000 U/mL 
solution to minimize adsorption of the drug to glass and plastic containers.
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32.1.3 Drug Administration
Interferon alpha-2b (dose-escalation from 0, 5, 10, and 20 MU/M2 administered as IV over 20 
minutes) will be administered intravenously on Day 1 and Day 3 (+ 1 day), as per the standard at 
Roswell.
32.1.4 Drug Storage and Accountability
The Investigator or designate will be responsible for ensuring that the investigational product is 
securely maintained in a locked, limited-access facility in accordance with the applicable 
regulatory requirements.
Powder for injection should be stored at 2 to 8°C (36-46°F). After reconstitution, the solution 
should be used immediately but may be stored up to 24 hours at 2-8°C (36-46°F).
Drug storage temperature will be maintained and recorded, as applicable.
32.1.5 Concomitant Medications
Interactions between interferon alpha-2b and other drugs have not been fully evaluated. Caution 
should be exercised when administering interferon alpha-2b therapy in combination with other 
potentially myelosuppressive agents. Various medications, such as cytotoxic chemotherapy and 
immunosuppressive regimens are likely to be discontinued as standard of care in patients with an 
acute viral infection. However, this protocol will not exclude patients based on recent or continued 
use of anti-cancer therapy. There is theoretical concern, but not a contraindication, regarding 
NSAIDs and selective COX-2 inhibitors in patients with COVID-19 infection; these agents won’t 
be offered as part of the study protocol, but are not excluded. Concomitant use of interferon alpha-
2b and theophylline decreases theophylline clearance, resulting in a 100% increase in serum 
theophylline levels. Concomitant interferon alpha-2b and REBETOL® (Ribavirin) use is 
contraindicated. A more detailed guide concerning drug interactions can be found in Appendix B.
32.2 Rintatolimod (poly IC analog)
A substituted double stranded poly-ribonucleic acid (polyI:polyC12U), rintatolimod preserves 
activity of poly-IC with a much-improved systemic toxicity profile. The product has been studied 
extensively for use as a vaccine adjuvant and for its direct antiviral activity, as well in several 
cancer studies as a monotherapy, but most extensively in chronic fatigue syndrome (CFS).
32.2.1 Other names
polyIC12U, Ampligen®, poly I: polyC12U; Polyinosinic: polycytidylic-polyuridylic acid; 
polyriboinosinic-polyribocytidylic (uridylic) acid.
32.2.2 Formulation and packaging
Rintatolimod is supplied as a liquid solution in glass bottles containing 200 mg (100 mg in case of 
toxicity) per 80 mL. Rintatolimod is a colorless solution containing 2.5 mg/mL in physiological 
salts (0.15 M NaCl, 0.01 M phosphate, 0.001 M Mg++). The product does not contain 
preservatives or antioxidants.
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32.2.3 Drug Shipment
Rintatolimod will be provided by AIM (previous name: Hemispherx) and shipped to the 
participating site.
The date of receipt and the amount of drug received will be documented. Drug shipment records 
will be retained by the investigational pharmacist or designee.
32.2.4 Preparing and Dispensing
A vial of rintatolimod is suitable for direct IV infusion. IDS will prepare and dispense. Each vial 
should be taken from the refrigerator and allowed to equilibrate to room temperature.
32.2.5 Drug administration
Rintatolimod 200 mg will be administered by intravenous infusion after interferon alpha-2b. 
Additional details on the procedures for receiving, storing and using rintatolimod (Ampligen®) 
can be found in a separate document entitled “Procedures for Receiving, Storing, and Using 
Ampligen® (Poly I:Poly C12U) Liquid Solution”.
The initial administration to start at 20 mL/ hour and increase to 40 mL/ hour after 30 minutes as 
per the standard at Roswell Total duration of rintatolimod infusion: 2.5 h-3h. Tubing should be 
flushed with 30 to 50 mL of normal saline solution upon completion. Administration will be 
followed by 1 hour of observation and vital signs at 30 and 60 minutes post-infusion (± 5 minutes).
32.2.6 Drug storage and accountability
The Investigator or designate will be responsible for ensuring that the investigational product is 
securely maintained in a locked, limited-access facility, as specified by Hemispherx and in 
accordance with the applicable regulatory requirements.
Rintatolimod should be stored at 2 to 8°C but should be infused at room temperature. Used vials 
should be accounted for and destroyed according to institutional procedure.
Drug storage temperature will be maintained and recorded, as applicable.
32.2.7 Handling and Disposal
The Investigator or designee will be responsible for dispensing and accounting for all 
investigational drug provided by Hemispherx exercising accepted medical and pharmaceutical 
practices. Study drugs must be handled as cytotoxic agents and appropriate precautions taken per 
the institution’s environmentally safe handling procedures. All investigational drugs will be 
dispensed in accordance with the Investigator’s prescription or written order.
All products dispensed will be recorded on a product accountability record. Records of product lot 
numbers and dates received will be entered on a product accountability form. This record will be 
reviewed by the Sponsor’s staff or representative during periodic monitoring visits. It is the 
Investigator’s responsibility to ensure that an accurate record of investigational drug issued and 
returned is maintained.
Used vials (excess drug) will be destroyed according to standard practices after properly 
accounting for the dispensing. Partially used vials of study drug will not be re-used for other 
participants.
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Under no circumstances will the Investigator supply investigational drug to a third party or allow 
the investigational drug to be used in a manner other than as directed by this protocol.
In regard to drug receipt, accountability and storage, SOP IDS-601 will be followed
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34 APPENDICES/ SUPPLEMENTS
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Appendix A
NYHA CLASSIFICATION
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Appendix B
DESCRIPTION OF INTRON® A POTENTIAL INTERACTIONS
Aldesleukin: Interferons (Alfa) may enhance the adverse/toxic effect of Aldesleukin. In 
particular, risks of myocardial and renal toxicity may be increased by this combination. 
Risk D: Consider therapy modification
Methadone: Interferons (Alfa) may increase the serum concentration of Methadone. Risk C: 
Monitor therapy
Ribavirin: Interferons (Alfa) may enhance the adverse/toxic effect of Ribavirin. Hemolytic 
anemia has been observed. Risk C: Monitor therapy
Telbivudine: Interferon Alfa-2b may enhance the adverse/toxic effect of Telbivudine. 
Specifically, the risk for peripheral neuropathy may be increased. Risk X: Avoid 
combination
Theophylline Derivatives: Interferons may decrease the metabolism of Theophylline 
Derivatives. Exceptions: Dyphylline. Risk C: Monitor therapy
Zidovudine: Interferons may enhance the adverse/toxic effect of Zidovudine. Interferons may 
decrease the metabolism of Zidovudine. Risk C: Monitor therapy
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Appendix C
ECOG Performance Status Scores
Description
Status
Fully active, able to carry on all pre-disease performance without restriction.
0
Restricted in physically strenuous activity but ambulatory and able to carry out work of a light 
or sedentary nature, e.g., light housework, office work.
1
Ambulatory and capable of all self-care but unable to carry out any work activities.
2
Capable of only limited self-care, confined to bed or chair more than 50% of waking hours.
3
Completely disabled. Cannot carry on any self-care. Totally confined to bed or chair.
4
Dead
5
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Appendix D
INVESTIGATOR STUDY ELIGIBILITY VERIFICATION FORM
INCLUSION CRITERIA (Main Cohort)
Participant Name: __________________________________________________
Medical Record No.: _________________________________________________
Title: Phase 1/2A Study of Rintatolimod and IFN alpha Regimen in Cancer Patients with COVID-
19 
Both men and women and members of all races and ethnic groups are eligible for this study.
INCLUSION CRITERIA (Main Cohort)
Yes
No
N/A
All answers must be “Yes” or “N/A” for participant 
enrollment.
Date
1.
Patients with cancer, with the exception of patients with 
active acute leukemia and allogeneic hematopoietic stem 
cell transplant recipients. Patients may be on active therapy 
or received therapy (e.g., chemotherapy, radiation, or 
surgery) within 7 years. Patients with active cancer who 
have not yet been treated (e.g. newly diagnosed cancer or 
early stage MDS or CLL) are eligible.  Basal cell cancer 
and carcinoma in situ treated with local excision alone do 
not qualify for inclusion.
2.
Presence of symptomatic infection, defined by fever (T ≥ 
38.0 ºC)  OR respiratory symptoms (cough, nasal 
congestion, or shortness of breath), OR lung infiltrates on 
chest X-ray or CT imaging, Diagnosis of COVID-19 based 
on PCR testing of respiratory samples. 
3.
Age equal to ≥ 18 years or older (children are excluded 
because COVID-19 typically has a milder course in 
children, and lack of safety data of this regimen in 
children).
4.
Laboratory evaluations:
o
Platelet ≥ 75,000/µL
o
Hemoglobin ≥ 9 g/dL
o
Hematocrit ≥ 27%
o
Absolute Neutrophil Count (ANC) ≥ 1000/µL
o
Creatinine clearance ≥ 50 mL/min (Cockroft-Gault 
Equation–note: plasma creatine instead of serum is used 
at Roswell Park)
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 57 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
INCLUSION CRITERIA (Main Cohort)
Yes
No
N/A
All answers must be “Yes” or “N/A” for participant 
enrollment.
Date
o
Total bilirubin ≤ 2 X institutional ULN 
o
AST (plasma) and ALT(plasma) ≤ 2 X institutional 
ULN
o
Plasma amylase and lipase ≤2 X institutional ULN
5.
In the absence of COVID-19, a life expectancy of at least 6 
months is expected.
6.
Participant or health care proxy must understand the 
investigational nature of this study and sign an Independent 
Ethics Committee/Institutional Review Board approved 
written informed consent form prior to receiving any study 
related procedure.
Investigator Signature: ___________________________________
Date: ___________
Printed Name of Investigator: __________________________________________________
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 58 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
INVESTIGATOR STUDY ELIGIBILITY VERIFICATION FORM
INCLUSION CRITERIA (Expansion Cohort)
Participant Name: __________________________________________________
Medical Record No.: _________________________________________________
Title: Phase 1/2A Study of Rintatolimod and IFN alpha Regimen in Cancer Patients COVID-19 
Both men and women and members of all races and ethnic groups are eligible for this study.
INCLUSION CRITERIA (Expansion Cohort)
Yes
No
N/A
All answers must be “Yes” or “N/A” for participant 
enrollment.
Date
1.
Patients with cancer or allogeneic stem cell transplant 
recipients with and without a cancer diagnosis.
o
Patients with cancer may be on active therapy or received 
therapy (e.g., chemotherapy, radiation, or surgery) within 
7 years.
o
Patients with active cancer who have not yet been treated 
(e.g. newly diagnosed cancer or early stage MDS or 
CLL) are eligible.
o
Basal cell cancer and carcinoma in situ treated with local 
excision alone do not qualify for inclusion.
2.
Presence of symptomatic infection, defined by fever 
(T≥38.0°C ) OR respiratory symptoms (cough, nasal 
congestion, or shortness of breath), OR lung infiltrates on 
chest X-ray or CT imaging.  Diagnosis of COVID-19 is 
based on PCR testing of respiratory samples. Severe 
infection is excluded (see Exclusion Criteria).
3.
Age equal to ≥ 18 years or older (children are excluded 
because COVID-19 typically has a milder course in 
children, and lack of safety data of this regimen in 
children). In the absence of COVID-19, a life expectancy of 
at least 6 months is expected.
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 59 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
INCLUSION CRITERIA (Expansion Cohort)
Yes
No
N/A
All answers must be “Yes” or “N/A” for participant 
enrollment.
Date
4.
Participant or health care proxy must understand the 
investigational nature of this study and sign an Independent 
Ethics Committee/Institutional Review Board approved 
written informed consent form prior to receiving any study 
related procedure.  There may be specific instances when the 
patient can’t provide informed consent, e.g. they require 
mechanical ventilation and are sedated, in which case a 
health care proxy will be able to provide informed consent. 
Patients with temporary cognitive impairment will be 
consented once their capacity has returned. Patients with 
chronic cognitive impairment, e.g. dementia, that precludes 
informed consent will not be enrolled.
Investigator Signature: ___________________________________
Date: ___________
Printed Name of Investigator: __________________________________________________
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 60 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
Appendix E
INVESTIGATOR STUDY ELIGIBILITY VERIFICATION FORM
EXCLUSION CRITERIA (Main Cohort)
Participant Name: _____________________________________________________
Medical Record No.:____________________________________________________
Title: Phase 1/2A Study of Rintatolimod and IFN alpha Regimen in Cancer Patients with COVID-
19 
EXCLUSION CRITERIA (Main Cohort)
Yes
No
N/A
All answers must be “No” or “N/A” for participant 
enrollment.
Date
1.
Patients with severe COVID-19 infection defined by 
pulmonary infiltrates on Chest X-Ray or CT imaging plus 
one of the following: room air SaO2 ≤ 92%, room air 
PaO2 <70 mm Hg, or PaO2-PaO2 ≥35 mm Hg.
2.
Contraindication to r-INFα based on prior hypersensitivity, 
autoimmune hepatitis, decompensated liver disease.
3.
Patients who have active acute myeloid leukemia or acute 
lymphoid leukemia or are allogeneic hematopoietic stem 
transplant recipients. Acute leukemia in remission and 
chronic leukemias are not exclusion criteria.
4.
Cardiac events:
a.
Acute coronary syndrome, myocardial infarction, or 
ischemia within past 3 months , or
b. New York Heart Association classification of III or IV 
congestive heart failure (Appendix A)
5.
Unwilling or unable to follow protocol requirements.
6.
Patients with known serious mood disorders.
7.
Any additional condition, such as pre-existing 
inflammatory lung disease, which in the Investigator’s 
opinion deems the participant an unsuitable candidate to 
receive the study drugs.
8.
Concurrent infections, e.g. bacterial pneumonia, or sepsis, 
that would make it difficult to evaluate clinical response to 
therapy or study drug toxicities.
9.
Therapies known to cause cytokine release syndrome 
(CRS), e.g. engineered T cells, within 30 days.
10. Patients at high risk for tumor lysis syndrome.
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 61 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
EXCLUSION CRITERIA (Main Cohort)
Yes
No
N/A
All answers must be “No” or “N/A” for participant 
enrollment.
Date
11. Concurrent active pneumonitis predating COVID-19, such 
as from checkpoint inhibitor therapy, chemotherapy-
associated toxicity, or radiation pneumonitis.
12. Autoimmune disease that requires systemic 
immunosuppression.
13. Protocol-defined baseline abnormalities in cell counts, 
renal, or hepatic function.
14. Any additional condition which in the Investigator’s 
opinion deems the participant an unsuitable candidate to 
receive the study drugs.
Participant meets all entry criteria:
  Yes
  No
If “NO”, do not enroll participant in study.
Investigator Signature: ___________________________________
Date: __________
Printed Name of Investigator: _________________________________________________
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 62 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
INVESTIGATOR STUDY ELIGIBILITY VERIFICATION FORM
EXCLUSION CRITERIA (Expansion Cohort)
Participant Name: _____________________________________________________
Medical Record No.:____________________________________________________
Title: Phase 1/2A Study of Rintatolimod and IFN alpha Regimen in Cancer Patients with COVID-
19 
EXCLUSION CRITERIA (Expansion Cohort)
Yes
No
N/A
All answers must be “No” or “N/A” for participant 
enrollment.
Date
1.
Patients with respiratory failure requiring mechanical 
ventilation with FIO2 of > 60%. 
2.
Allogeneic hematopoietic stem cell transplant recipients 
with pulmonary GvHD (any grade) 
3.
Cardiac events: 
a.
Acute coronary syndrome, myocardial infarction, or 
ischemia within past 3 months , or
b. New York Heart Association classification of III or IV 
congestive heart failure (Appendix A)
4.
Unwilling or unable to follow protocol requirements.
5.
Any additional condition which in the Investigator’s 
opinion deems the participant an unsuitable candidate to 
receive the study drugs.
Participant meets all entry criteria:
  Yes
  No
If “NO”, do not enroll participant in study.
Investigator Signature: ___________________________________
Date: __________
Printed Name of Investigator: _________________________________________________
APPROVED RPCI IRB
3/6/2025

Roswell Park Protocol No.: I-659920
Page 63 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
Appendix F
Study Calendar (Main Cohort)
Baseline and/or Screening assessments must be performed within 3 days prior to the first dose of investigational product unless otherwise 
stated. Therapy may begin on the same day as screening. If screening and day 1 are completed on the same day, then assessments 
required on both days may be done only once*. Any results falling outside of the reference ranges may be repeated at the discretion of 
the investigator. 
*If screening and day 1 occur on the same day, correlative blood draw and nasopharyngeal swab collection will be as follows:
Correlative blood draw: In the case when day one of treatment is the same as the screening day, only two blood draws (30 mL each 
time) are needed for the correlative studies on that day: Before the start of treatment and within 1 hour after the end of the combined 
infusion (i.e., collect one pre-dose sample and one sample within one hour after end of rintatolimod IV, if screening and Day 1 are the 
same day).
Nasopharyngeal swabs: In the case when day one of treatment is the same as the screening day, only one nasopharyngeal swab (pre-
dose) is required.
Evaluation
Screening and 
Informed Consent
Day 1
Day 3
(+ 1 day)
Day 7 (± 1 day) and
Day 14 (± 3 days)5 
Written Informed Consent prior to any 
Screening Procedures
X
Recording of Concomitant 
Medications
X
X
X
Recording of Adverse Events
X
X
X
Survival Follow-up (through Day 14)
X
X
X
Clinical Assessments
Medical & Surgical History
X

Roswell Park Protocol No.: I-659920
Page 64 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
Evaluation
Screening and 
Informed Consent
Day 1
Day 3
(+ 1 day)
Day 7 (± 1 day) and
Day 14 (± 3 days)5 
Physical Exam including vital signs3 
X
X
X
X
ECOG Performance Status
X
X
X
X
Laboratory Procedures
CBC with diff
X
X
X
X
CMP
X
X
X
X
Serum hCG in women of child-
bearing potential
X
ABG
X
Blood (30cc) for immunologic 
studies2
X
X (prior to treatment)
and
X (within 1hr after end 
of infusion treatment)
X (prior to 
treatment)
and
X (within 1hr after 
end of infusion 
treatment)
X2a
Nasopharyngeal Swab (as required 
and based on availability)
X
X
X
X
Imaging/
Other Procedures
12-Lead ECG
X
CXR or CT scan4
X

Roswell Park Protocol No.: I-659920
Page 65 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
Evaluation
Screening and 
Informed Consent
Day 1
Day 3
(+ 1 day)
Day 7 (± 1 day) and
Day 14 (± 3 days)5 
Clinical evaluation of endpoints1
X
X
X
X
Study Drug Administration
Rintatolimod/IFN regimen
Administration + best available SOC 
X
X
1. Hospitalization in patients who were not inpatients at study entry; respiratory failure requiring mechanical ventilation; 14-day 
survival.
2. Blood (30cc) for correlative studies: On Day 1 and Day 3 prior to treatment and within 1 hour after end of infusion treatment 
(two draws on each of these days). Note: The combination treatment begins with IV IFNα followed by IV rintatolimod, with 
the correlative blood draw within 1 hour following IV rintatolimod as indicated above. Refer to Section 11.4. Single draws on 
the other days.
2a: Follow-up visits- NOTE: In the event patient cannot tolerate treatment with all 2 days of treatment, then blood (30cc) for 
correlatives will be collected at time that the patient recovers from toxicities (ideally within 1 week of the last infusion).
3. Physical exam will be limited to vitals and auscultation of lungs. Vital signs (temperature, heart rate, respiratory rate, blood 
pressure, SaO2), body weight, and height: Height collected at baseline only.
4. NOTE: If a chest CT is ordered as standard of care prior to starting treatment, a chest x-ray is not required.
5. While every effort will be made to complete assessments, there are limitations for outpatients, including securing transport to 
Roswell Park. Some follow up visits will need to be done virtually, and in other cases, may be limited to viral swabs. These 
missed assessments or sample collections for outpatients will not be considered protocol violations.

Roswell Park Protocol No.: I-659920
Page 66 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
Study Calendar (Expansion Basket: Rintatolimod) 
Evaluation
Screening and 
Informed Consent
Day 1
Day 3 (or Day 4)5
Day 7 (± 1 day) and
Day 14 (± 3 days) and
Day 30 (+ 3 days)6,7
Written Informed Consent 
prior to any Screening 
Procedures
X
Recording of Concomitant 
Medications
X
X
X
Recording of Adverse 
Events
X
X
X
Survival Follow-up 
(through Day 14)
X
X
X
Clinical Assessments
Medical & Surgical 
History
X
Physical Exam including 
vital signs3 
X
X
X
X
ECOG Performance Status
X
X
X
X
Laboratory Procedures
CBC with diff
X
X
X
X
CMP
X
X
X
X
Serum hCG in women of 
child-bearing potential
X

Roswell Park Protocol No.: I-659920
Page 67 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
Evaluation
Screening and 
Informed Consent
Day 1
Day 3 (or Day 4)5
Day 7 (± 1 day) and
Day 14 (± 3 days) and
Day 30 (+ 3 days)6,7
ABG
X
Blood (30cc) for 
immunologic studies2
X
X (prior to treatment)
and
X (within 1hr after 
end of infusion 
treatment)
X
X2a
Nasopharyngeal Swab (as 
required and based on 
availability)
X
X
X
X
Imaging/
Other Procedures
12-Lead ECG
X
CXR or CT scan4
X
Clinical evaluation of 
endpoints1
X
X
X
X
Study Drug 
Administration
Rintatolimod
Administration + best 
available SOC 
X
X
X6
1. Hospitalization in patients who were not inpatients at study entry; respiratory failure requiring mechanical ventilation; 15-day 
survival.
2. Blood (30cc) for correlative studies: On Day 1 prior to treatment and within 1 hour after end of infusion treatment (two draws 
on Day 1). Note: The treatment begins with IV rintatolimod, with the correlative blood draw within 1 hour following IV 
rintatolimod as indicated above. Refer to Section 11.4. 

Roswell Park Protocol No.: I-659920
Page 68 of 68
Date: 02/13/2025; 3/6/2025 (IRB)
2a: Follow-up visits- NOTE: In the event patient cannot tolerate treatment, then blood (30cc) for correlatives will be collected 
at time that the patient recovers from toxicities (ideally within 1 week of the infusion).
3. Physical exam will be limited to vitals and auscultation of lungs. Vital signs (temperature, heart rate, respiratory rate, blood 
pressure, SaO2), body weight, and height: Height collected at baseline only.
4. NOTE: If a chest CT is ordered as standard of care prior to starting treatment, a chest x-ray is not required.
5. Based on the PI’s judgment, additional infusions of  rintatolimod at 200 mg may be administered 3 to 4 days apart, after the 
Day 3 (or Day 4) administration. Decisions about additional rintatolimod infusions will be based on evaluation of AEs 
attributed to rintatolimod, particularly increased FIO2 needs. Patients who do not receive additional rintatolimod infusions will 
follow the above schedule for EOT/Follow-up visit.  
6. Based on the PI’s judgment, additional infusions of  rintatolimod at 200 mg may be administered 3 to 4 days apart. Decisions 
about additional rintatolimod infusions will be based on evaluation of AEs attributed to rintatolimod, particularly increased 
FIO2 needs. A maximum of 12 rintatolimod infusions may be administered per patient. 
7. While every effort will be made to complete assessments, there are limitations for outpatients, including securing transport to 
Roswell Park. Some follow up visits will need to be done virtually, and in other cases, may be limited to viral swabs. These 
missed assessments or sample collections for outpatients will not be considered protocol violations.