Phase I/II Study of High Dose Interleukin 2, Aldesleukin, in Combination With the Histone Deacetylase Inhibitor Entinostat in Patients With Metastatic Renal Cell Carcinoma
Phase: PHASE1
Conditions: Clear Cell Renal Cell Carcinoma, Metastatic Kidney Carcinoma, Stage III Renal Cell Cancer AJCC v7, Stage IV Renal Cell Cancer AJCC v7
Sponsor: National Cancer Institute (NCI)

 
page I 
PROTOCOL SUMMARY OF CHANGES 
RPCI # I 145208 / NCI 7870 
Amendment: RPCI #31 / CTEP #32 Protocol Dated: 5/29/19 
Study Title: Phase I / II Study of High Dose Interleukin 2, Aldesleukin, in Combination with the Histone Deacetylase Inhibitor Entinostat in Patients 
with Metastatic Renal Cell Carcinoma 
 
Section 
Page 
Rationale 
Previous Text 
New Text 
 
Title 
Page  
 
1 
 
 
 
Updated for 
submission 
 
 
Amendment # 30 10/11/18 (CTEP version 10/9/18 
Amendment # 31) 
 
Amendment # 31 05/29/19 (CTEP version 10/9/18 
Amendment # 32) 
Footer 
All 
Updated for 
Submission 
10/11/18 
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4.3 
17 
Updated 
to 
add 
in 
termination 
language 
once primary 
endpoint 
is 
met. 
Duration of Follow Up 
 
Follow up safety evaluations will occur 30 days 
(+/- 4 days) after last dose of study drug or until 
resolution of any drug related toxicity (telephone 
contact is acceptable). Follow up will include 
review of concomitant medications and adverse 
events. 
 
 
Patients (both Phase I and Phase II) will be 
followed up after completion of therapy every 
three months for survival assessment (telephone 
contact is acceptable). 
Duration of Follow Up 
 
Follow up safety evaluations will occur 30 days (+/- 4 
days) after last dose of study drug or until resolution 
of any drug related toxicity (telephone contact is 
acceptable). Follow up will include review of 
concomitant medications and adverse events. 
 
 
Patients (both Phase I and Phase II) will be followed 
up after completion of therapy every three months for 
survival assessment (telephone contact is acceptable) 
or until completion of primary endpoint/ termination 
of study. 
ICF 
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Addendum 
10/11/18 
5/29/19 

Entinostat and Aldesleukin 
                           PI: Saby George, M.D 
 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
 page 1 of 78 
 
 
Title: Phase I/II Study of High Dose Interleukin 2, Aldesleukin, in 
Combination with the Histone Deacetylase Inhibitor Entinostat in Patients 
with Metastatic Renal Cell Carcinoma 
 
RPCI Study No.: 
 
I 145208 
 
 
 
Original Version: 11-7-08 
 
 
 
Amendment # 1, 4-14-09 (Includes up to CTEP amendment # 11) 
 
 
 
Amendment # 2 10-15-09 
 
 
 
Amendment # 3 10-23-09 
 
 
 
Amendment # 4 11-27-09 
 
 
 
Amendment # 5 1-13-10 (CTEP version 1-14-2010) 
 
 
 
Amendment # 6 3-4-10 (CTEP version 3-4-10) 
 
 
 
Amendment # 7 5-7-10 (CTEP version 5-7-10 amendment #4) 
 
 
 
Amendment # 8 9/1/10 (CTEP version 9/1/10 amendment #5) 
 
 
 
Amendment # 9 12/16/10 (CTEP version 12/16/10 amendment # 6) 
Amendment #10 4/2/11 (CTEP amendment #7withdrawn) 
 
 
 
Amendment #11 6/21/11(CTEP version 6/21/11 amendment #8) 
Amendment #11 7/15/11 (CTEP version 7/15/11 amendment #9) 
Amendment #12 9/13/11 (CTEP version 9/13/11 amendment #10) 
Amendment #13 10/3/11 (CTEP version 10/3/11 amendment #11) 
Amendment #13 11/16/11(CTEP version 11/16/11 amendment #12) 
Amendment #14 5/23//12 (CTEP version 5/23/12 amendment #13) 
Amendment #14 5/23/12 (CTEP version 6/5/12 amendment #14) 
Amendment #15 11/27/12 (CTEP version 11/27/12 amendment #15) 
Amendment # 16 3/12/13 (CTEP version 3/12/13 amendment #16) 
Amendment #17 5/29/13 (CTEP version 5/29/13 amendment #17) 
Amendment #17 6/25/13 (CTEP version 6/25/13 amendment #18) 
Amendment #18 12/5/13 (CTEP version 12/5/13 amendment # 19) 
Amendment # 19 2/10/14 (CTEP version 2/10/14 amendment #20) 
Amendment #19 3/13/14 (CTEP version 3/13/14 amendment #21) 
Amendment #20 4/22/14 (CTEP version 4/22/14 amendment #22 
                                           Amendment #21 6/11/14 (CTEP version 6/11/14 amendment #23) 
 
 
              Amendment #22 10/22/14(CTEP version 10/22/14 amendment #24) 
 
 
 
Amendment #23 03/17/15(CTEP version 03/17/15amendment #25) 
 
 
 
Amendment #24 08/20/15 (CTEP version 08/20/15 amendment #26) 
 
 
 
Amendment #25 09/04/15 (CTEP version 09/04/15 amendment #27) 
 
 
 
Amendment #26 11/16/15 (CTEP version 11/16/15 amendment #28) 
 
 
 
Amendment #27 1/27/16 (CTEP version 1/27/16 amendment #29) 
Amendment # 28 3/2/15 (CTEP version 3/2/16 Amendment # 30) 
Amendment # 29 4/14/17 (CTEP version 3/2/16 Amendment # 30) 
Amendment #30 10/11/18 (CTEP version 10/9/18 Amendment #31) 
Amendment #31 5/29/19 (CTEP version 10/9/18 Amendment #32) 
 
Coordinating Site: 
 
Roswell Park Cancer Institute 
Elm & Carlton Streets 
Buffalo, NY 14263-0001 
 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 2 of 78 
 
Principal Investigator: 
Saby George, MD 
 
 
 
 
Roswell Park Cancer Institute 
 
 
 
 
Elm & Carlton Streets 
Buffalo, New York 14263 
Tel 716-845-3159 
Fax 716-845-8346 
Email Saby.George@RoswellPark.org 
 
Co-Investigators: 
Amy Early, MD 
 
 
 
  
 
 
Tel 716-845-7610 
 
 
 
  
        
Fax: 716-845-2949 
 
Email:Amy.Early @RoswellPark.org 
 
 
  
 
 
 
Gissou Azabdaftari, MD (no patient care) 
Tel 716-845-3867 
Email : Gissou.Azabdaftari@RoswellPark.org 
 
Network Sites: #1) The Sidney Kimmel Comprehensive Cancer Center (SKCCC) 
at Johns Hopkins University (JHU) 
Bunting-Blaustein Cancer Research Building 
1650 Orleans Street, Room 1M52 
Baltimore, MD 21231-1000 
 
Principal Investigator: 
Michael Anthony Carducci, MD 
Co-Investigators: 
Hans Hammers MD PhD, Medical Oncology 
Michelle Rudek, Pharm.D, PhD (no patient care) 
Ephraim Fuchs MD, PhD Medical Oncology (no patient care) 
Mario Eisenberger, M.D. Medical Oncology 
Samuel Denmeade, M.D.Medical Oncology 
Janet Walczak, NP Medical Oncology 
Victoria Sinibaldi, NP Medical Oncology 
 
Martin Lodge PhD, Radiology (no patient care) 
Jenny Kim, MD  
 
#2) The Ohio State University 
A433A Starling-Loving Hall 320 W 10th Ave  
Columbus, OH 4321 
 
Principal Investigator:  
J. Paul Monk III, MD 
Co-Investigators: 
Arvinder S. Bhinder, MD 
Amir Mortazavi, MD 
Steven K. Clinton, MD, PhD 
Thomas Olencki, MD 
 
#3) USC/Norris Comprehensive Cancer Center and Hospital 
1441 Eastlake Avenue  
Los Angeles, CA 90033-2211 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 3 of 78 
 
Principal Investigator:  
David Quinn MBBS, PhD, FRACP 
Co-Investigators: 
Tanya Dorff, MD 
James Hu, MD 
Michael Wong, MD 
 
NCI Supplied Agent: Entinostat (MS-275, SNDX-275), NSC 706995, IND 61198 
 
 
 
 
 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 4 of 78 
 
SCHEMA 
 
Metastatic renal cell carcinoma patients meeting eligibility criteria in section 3.0 
 
REGISTRATION 
 
 
Entinostat: at assigned dose PO every 2 weeks 
High dose aldesleukin: 600,000 IU/kg IV q 8 hours, day 1-5 and 15-19 (± 7 days) 
1 cycle = 84 days 
 
 
 
Evaluate q cycle (per section 4.0) 
 
 
 
Disease progression 
Unacceptable toxicity 
Patient decision to discontinue study drugs 
Alternate treatment 
 
Event-monitoring (see section 10.0) 
____________________________ 
 
Drug Names/Abbreviations 
 
 
Generic name: Entinostat  
Generic name: Interleukin 2 
Brand name: Aldesleukin/Proleukin 
 
 
 
 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 5 of 78 
 
TABLE OF CONTENTS 
Page 
TITLE: PHASE I/II STUDY OF HIGH DOSE INTERLEUKIN 2, ALDESLEUKIN, IN 
COMBINATION WITH THE HISTONE DEACETYLASE INHIBITOR ENTINOSTAT 
IN PATIENTS WITH METASTATIC RENAL CELL CARCINOMA ................................. 1 
RPCI STUDY NO.:  I 145208 .................................................................................................... 1 
PRINCIPAL INVESTIGATOR: SABY GEORGE, MD ....................................................... 2 
CO-INVESTIGATORS:............................................................................................................... 2 
SCHEMA ....................................................................................................................................... 4 
1. 
OBJECTIVES ....................................................................................................................... 8 
1.1 
Phase I Primary Objectives ..............................................................................................8 
1.2 
Phase II Primary Objectives ............................................................................................8 
1.3 
Phase II Secondary Objectives ........................................................................................8 
2. 
BACKGROUND ................................................................................................................... 8 
2.1 
Renal cell carcinoma ........................................................................................................8 
2.2 
Interleukin 2 (Aldesleukin) ..............................................................................................8 
2.3 
Histone deacetylase inhibitors .........................................................................................9 
2.4 
Entinostat .......................................................................................................................10 
2.5 
Rationale for the combination of entinostat and high dose aldesleukin ........................10 
2.6 
Rationale for entinostat dose and schedule selection ....................................................12 
3. 
PATIENT SELECTION .................................................................................................... 12 
3.1 
Eligibility Criteria ..........................................................................................................12 
3.2 
Exclusion Criteria ..........................................................................................................14 
3.3 
Inclusion of Women and Minorities ..............................................................................15 
4. 
TREATMENT PLAN ......................................................................................................... 15 
4.1 
Entinostat / aldesleukin administration and definition of Dose-Limiting Toxicity (DLT)
 
16 
4.2 
Duration of Therapy ......................................................................................................17 
4.3 
Duration of Follow Up ...................................................................................................17 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 6 of 78 
 
5. 
DOSING DELAYS/DOSE MODIFICATIONS ............................................................... 17 
5.1 
Entinostat Treatment Modification for Adverse Events ................................................18 
5.2 
Adverse Events with entinostat .....................................................................................18 
5.3 
Aldesleukin Treatment Modification for Adverse Events .............................................19 
5.4 
Specific aldesleukin Toxicity Management ...................................................................20 
5.5 
Management of Grade 4 Toxicity ..................................................................................21 
5.6 
Toxicity Criteria for Discontinuing Treatment ..............................................................22 
6. 
PHARMACEUTICAL INFORMATION ......................................................................... 22 
6.1 
Entinostat (NSC 706995) ...............................................................................................22 
6.2 
Interleukin-2 (Aldesleukin) ...........................................................................................29 
7. 
CORRELATIVE/SPECIAL STUDIES ............................................................................ 30 
7.1 
Pharmacodynamic Studies .............................................................................................30 
7.2 
Pharmacokinetic studies (Phase I only) .........................................................................31 
7.3 
Non invasive imaging studies ........................................................................................32 
7.4 
Optional tumor Samples ................................................................................................32 
8. 
STUDY CALENDAR ......................................................................................................... 35 
9. 
MEASUREMENT OF EFFECT ....................................................................................... 37 
9.1 
Methods of Measurement ..............................................................................................37 
9.2 
Response Criteria ...........................................................................................................38 
10 REGULATORY AND REPORTING REQUIREMENTS ............................................. 41 
10.1 
Protocol Amendments ...................................................................................................41 
10.2 
Informed Consent ..........................................................................................................41 
10.3 
Adverse Events ..............................................................................................................42 
10.4 
Data and Safety Monitoring ...........................................................................................48 
10.5 
CTEP Multicenter Guidelines ........................................................................................48 
11. 
STATISTICAL CONSIDERATIONS .......................................................................... 51 
11.1 
Study Design- ................................................................................................................51 
11.2 
Phase I ............................................................................................................................51 
11.3 
Phase II ..........................................................................................................................52 
11.4 
Toxicity monitoring .......................................................................................................54 
11.5 
Patient Compliance ........................................................................................................55 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 7 of 78 
 
11.6 
Data quality assurance ...................................................................................................56 
11.7 
Reporting and Exclusion ................................................................................................57 
12. 
ETHICS ........................................................................................................................... 58 
12.1 
Institutional Review Board ............................................................................................58 
12.2 
Evaluation of benefits and risks/discomforts .................................................................58 
12.3 
Risks/benefits analysis ...................................................................................................58 
12.4 
Patient information and consent ....................................................................................58 
12.5 
Data handling and record keeping .................................................................................59 
13. 
REFERENCES ................................................................................................................ 61 
APPENDIX A: PROTOCOL FOR QUANTITATIVE ANALYSIS OF TUMOR 
METABOLISM AND TUMOR BLOOD FLOW USING 18FDG–PET/CT IMAGING. ... 63 
APPENDIX B: PERFORMANCE STATUS CRITERIA ....................................................... 67 
APPENDIX 
C: 
HDIL-2 
(HIGH 
DOSE 
ALDESLEUKIN) 
ADMINISTRATION 
GUIDELINES ............................................................................................................................. 68 
APPENDIX D: INSTRUCTIONS FOR NETWORK SITES ................................................. 77 
 
 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 8 of 78 
 
1. OBJECTIVES 
 
1.1 
Phase I Primary Objectives 
1.1.1 To evaluate the safety and tolerability of high dose interleukin 2 (aldesleukin) in 
combination with entinostat in patients with metastatic renal cell carcinoma (RCC). 
 
1.2 
Phase II Primary Objectives 
1.2.1 To monitor toxicity and estimate the efficacy of high dose aldesleukin combined 
with entinostat in patients with metastatic RCC. 
 
1.3 
Phase II Secondary Objectives 
1.3.1 To compare the time-to-tumor progression, progression-free survival and overall 
survival of patients with metastatic RCC treated with high dose aldesleukin 
combined with entinostat to the historical data of patients treated with high dose 
aldesleukin alone. 
1.3.2 To assess the toxicity of high dose aldesleukin combined with entinostat. 
1.3.3 To evaluate entinostat pharmacodynamics (PD) in blood and tumor samples. 
1.3.4 To measure the association between baseline laboratory parameters (e.g. CD4+, 
CD8+, CD4+/Foxp3), tumor blood metabolism, and a variety of response variables 
(e.g. toxicity, response and survival). 
1.3.5 To explore the relationship between entinostat exposure with PD endpoints (e.g. 
toxicity and histone acetylation in peripheral blood mononuclear cells or PBMNCs 
and changes in T cell subset population). 
1.3.6 To evaluate the modulation of tumor metabolism by FDG PET/CT scan. 
 
2. BACKGROUND 
 
2.1 
Renal cell carcinoma  
More than 51,000 new cases of kidney cancer and approximately 13,000 cancer specific deaths 
in the United States have been predicted for 20071. One third of patients have metastatic disease 
at the time of diagnosis. Two randomized studies in metastatic RCC patients have shown 
survival benefit of nephrectomy and cytokine therapy over cytokine therapy alone. Many 
patients undergo nephrectomy as a component of their standard care. The prognosis for recurrent 
or metastatic renal cell carcinoma is poor, as median survival is 10-13 months and 5-year 
survival is less than 5%. These figures underscore the need for effective systemic therapy in this 
disease. 
 
In 1992, high-dose bolus aldesleukin was approved by the FDA for the treatment of patients with 
metastatic renal cell cancer based on data presented on 255 patients entered into 7 phase II 
clinical trials2-4. HD bolus aldesleukin is superior in terms of response rate and response quality 
to regimens involving either intermediate or low dose aldesleukin or subcutaneous interferon 
alpha (IFN)5, 6. 
 
2.2 
Interleukin 2 (Aldesleukin) 
The Cytokine Working Group performed a phase III trial in which patients were randomized to 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 9 of 78 
 
receive either outpatient aldesleukin (5 MIU/m2 subcutaneously every 8 hours x 3 doses on day 1 
then daily 5 days/week for 4 weeks) and IFN (5 MIU/m2 subcutaneously thrice weekly for 4 
weeks) every 6 weeks or standard high dose inpatient aldesleukin (600,000 IU/kg/dose every 8 
hours intravenously, days 1-5 and 15-19 [max 28 doses]) every 12 weeks5. The response rate for 
high-dose aldesleukin was 23% (22/95) versus 10% (9/91) for aldesleukin / IFN (p=0.018). 
Eight patients achieved a complete response on high dose aldesleukin versus only 3 on low dose 
aldesleukin / IFN. The median response durations were 24 months for high dose aldesleukin 
and 15 months for aldesleukin / IFN (p=0.18). Median overall survivals were 17 and 13 months 
(p=0.21), favoring high dose aldesleukin. Ten patients on high dose aldesleukin were progression 
free at 3 years versus 3 on aldesleukin / IFN (p=0.08). Durable CRs favored HD aldesleukin (7 
vs. 0). Median progression free survival was 3 months for each treatment arm. Another three-arm 
randomized study compared response rates and overall survival for patients with metastatic RCC 
receiving high-dose or one of two low-dose aldesleukin regimens7. Major tumor regressions, as 
well as complete responses, were seen with all regimens tested but high dose aldesleukin was 
more clinically active, although this did not produce an overall survival benefit. Taken together 
these data suggests that high dose aldesleukin should remain the preferred therapy for 
appropriately selected patients with access to such therapy before or after the use of the recently 
FDA approved receptor tyrosine kinase inhibitors sorafenib and sunitinib8,9. However, given the 
toxicity and limited efficacy of high dose aldesleukin therapy, additional efforts should be 
directed to increase the efficacy of immunotherapy. 
 
2.3 
Histone deacetylase inhibitors 
 
Histone deacetylases (HDACs) are critically important in the regulation of gene expression and 
in the field of target-specific anticancer drug.10-12 Inhibitors of HDAC present an exciting, novel 
approach to the treatment of solid tumors, many of which are refractory to current therapies. 
Several chemical groups of HDAC inhibitors have been identified: 1) short-chain fatty acids – 
butyrates 2) hydroxamic acids – trichostatin A (TSA), suberoylanilide hydroxamic acid (SAHA 
or vorinostat), oxamflatin and LBH-589; 3) cyclic tetrapeptides containing a 2-amino-8-ox.o-
9,10-epoxy-decanoyl (AOE) moiety – trapoxin A: 4) cyclic peptides not containing the AOE 
moiety – depsipeptide and apicidin and 5) benzamides – entinostat. 
 
The HDACs exert their targeted action during post-translational acetylation of core nucleosomal 
histones, which affects chromatin structure, thereby regulating gene expression. DNA that is 
wrapped around condensed, non-acetylated histones is transcriptionally inactive, whereas 
acetylation of N-terminal histone lysine residues exposes DNA to important transcription factors 
that promote transcriptional activity. The dynamic equilibrium between histone acetylation and 
deacetylation is regulated by histone acetyltransferases (HATs) and HDACs. The action of 
HDACs on nucleosomal histones leads to tight coiling of chromatin and silencing of expression 
of various genes, including those implicated in the regulation of cell survival, proliferation, 
differentiation, and apoptosis. The effects of HDACs are not limited to histone deacetylation. 
HDACs also act as members of a protein complex to recruit transcription factors to the promoter 
region of genes, including those of tumor suppressors, and they affect the acetylation status of 
specific cell cycle regulatory proteins. 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 10 of 78 
 
2.4 
Entinostat 
 
The synthetic benzamide, entinostat has demonstrated induction of chromatin hyper-acetylation 
and antitumor activity by inhibition of HDAC enzyme activity. Entinostat has also demonstrated 
inhibition of tumor cell growth in nude mice that was comparable or superior to conventional 
cytotoxic agents13. The results from the Phase I testing of entinostat have been recently 
published14. Patients with advanced solid tumors or lymphoma were treated with entinostat 
orally initially on a once daily x 28 every 6 weeks (daily) and later on once every-14-days (q14-
day) schedules. The starting dose was 2 mg/m2 and the dose was escalated in three- to six-
patient cohorts based on toxicity assessments. With the daily schedule, the MTD was exceeded at 
the first dose level. PK analysis suggested the half-life of entinostat in humans was 39 to 80 
hours, substantially longer than predicted by preclinical studies. With the q14-day schedule, 28 
patients were treated. The MTD was 10 mg/m2 and dose-limiting toxicities were nausea, 
vomiting, anorexia, and fatigue. Exposure to entinostat was dose dependent, suggesting linear 
PK. Increased histone H3 acetylation in peripheral-blood mononuclear-cells was apparent at all 
dose levels by immunofluorescence analysis. Ten of 29 patients remained on treatment for > or = 
3 months. The entinostat oral formulation on the daily schedule was intolerable at a dose and 
schedule explored. The q14-day schedule is reasonably well tolerated. Histone deacetylase 
inhibition was observed in peripheral-blood mononuclear-cells. Based on PK data from the q14-
day schedule, more frequent dosing schedules such as weekly x 4, repeated every 6 weeks have 
been tested. In a Phase I study (Berlex 306121) the MTD for entinostat dosed on D1, D8, D15 of 
4 weeks cycle was established at 4mg/m2 dose level. Most common toxicities hypophosphatemia 
(grade 3 in 48% of patients) bone marrow suppression, gastrointestinal events such as nausea and 
vomiting (grade 1 + 2 frequent) fatigue, asthenia (grade 1 +2 frequent, hypoalbuminemia, 
hypoproteinemia. 
 
2.5 
Rationale for the combination of entinostat and high dose aldesleukin 
 
HDAC inhibitors induce cell-cycle arrest, differentiation or apoptosis in vitro, and have potent 
anti-tumor activities in vivo. We have previously showed HDAC inhibitor entinostat has a 
significant antitumor activity in a renal cell carcinoma model15. HDAC inhibitors have been also 
shown to have immunodulatory properties including activation of major histocompatibility 
complex (MHC) class I and II proteins, and co-stimulatory molecules CD40, CD80 and CD86. 
The results from the clinical trials with HDAC inhibitors in cutaneous T cell lymphoma and 
large cell lymphoma patients suggest that the antitumor activity these agents may be in part due 
to the modulation of the immune response. 
 
In our lab, we have recently tested the effectiveness of combination therapy of aldesleukin with 
the HDAC inhibitor entinostat in a murine renal cell carcinoma (RENCA) model16. RENCA-
luciferase expressing cells were implanted in the left kidney of BALB/C mice. Animals were 
randomly divided in four groups and treated with either vehicle, 150,000 IU aldesleukin twice 
daily by i.p injections (twice weekly for two weeks), 5 mg/kg entinostat daily by oral gavage (5 
days/week for two weeks), or combination. Treatment was started 3 days following tumor cell 
injection. Weekly luciferase images and tumor weight after 3 weeks treatment showed 
significant tumor inhibition (>80%) in the as compared to aldesleukin (no significant inhibition) 
or entinostat (~40% inhibition) (see Panel A and B). Spontaneous lung metastases were also 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 11 of 78 
 
inhibited in the combination treatment (>90 % inhibition) as compared to single treatment. 
Kaplan-Meier analyses showed statistically significant increase in survival in the combination 
group as compared with control and single agents (Panel C). The percentage of CD4+ 25+ and 
Fox-p3+ T cells was increased or reduced, respectively, in lymph nodes from tumor bearing 
animals treated with the combination of entinostat and aldesleukin as compared to control and 
single agents (Panel D). Splenocytes from mice treated with combination treatment showed 
greater lysis than splenocytes from mice treated with single agents (Panel E).These results 
suggest that combination of aldesleukin and entinostat has a synergistic antitumor effect in vivo 
in an immunocompetent murine model of renal cell carcinoma. The antitumor effect was 
associated with decreased number of regulatory T cells and increased antitumor cytotoxicity by 
splenocytes. Lowe dose aldesleukin (15,000 IU) also induced a greater antitumor effect as 
compared to single agents (Panel F). However, microscopic examination revealed the presence 
of tumor in all the samples suggesting a dose –dependent effect of aldesleukin in this 
combination (Panel G). Blood samples collected after two hours from entinostat administration 
revealed a median plasma concentration of 20.6 ±5.01 ng/ml. 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
In summary, these results suggest that combination of aldesleukin and entinostat has a 
synergistic antitumor effect in vivo in an immunocompetent murine model of renal cell 
carcinoma. The antitumor effect was associated with decreased number of regulatory T cells and 
increased antitumor cytotoxicity by splenocytes. A dose dependent effect for aldesleukin but not 
for entinostat was observed. In conclusion, these data provide the rationale for clinical testing of 
the combination of high dose aldesleukin and HDAC inhibitors in the treatment of renal cell 
carcinoma patients. The immunomodulatory activity of entinostat and its direct antitumor effect 
may increase the response rate to high dose aldesleukin, further delay disease progression and 
increase progression-free survival in patients with metastatic RCC. 
 
A
CTL
MS-275
IL-2
MS-275+IL-2
CTL
MS-275
IL-2
MS-275+IL-2
A 
A 
Prevention model
0.00E+00
5.00E+08
1.00E+09
1.50E+09
2.00E+09
2.50E+09
3.00E+09
3.50E+09
4.00E+09
4.50E+09
5.00E+09
day 3
day 10
day 17
day 24
CTL
IL2
MS275
Comb
Days after kidney injection
0
0.5
1
1.5
2
2.5
CTL
g
0
0.5
1
1.5
2
2.5
CTL
g
Tumor weight (g)
CTL
I
0
.2
.4
.6
.8
1
% survival
0
10
20
30
40
50
days
MS-275
IL-2
CTL
MS-275+IL-2
MS-275
IL-2
CTL
MS-275+IL-2
**
B
C 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 12 of 78 
 
2.6 
Rationale for entinostat dose and schedule selection 
 
Entinostat has been evaluated in vitro, in non-clinical in vivo studies, and in phase 1 studies in 
patients with various solid tumors and hematological malignancies at doses between 2 and 12 
mg/m2 and at dosing frequencies ranging from once daily to every 2 weeks. Increased histone 
acetylation was observed at the lowest dose evaluated with the effect persisting at least 48 hours 
post-dose. Pharmacokinetic studies of entinostat have indicated a long half-life of entinostat, 
Patients may have received up to two prior therapies including VEGF, mTOR and PD-
1/PDL1 inhibitors ranging from 40 hours to 120 hours, with concentrations detectable 168 
hours post-dose at doses of 6 mg/m2 to 12 mg/m2, consistent with this long half-life.  
 
The MTD for single agent in non-hematologic indications has been established as 4mg/m² 
weekly x 3 and one week rest, or 10 mg/m² every second week continuously. In combination 
with 13-cis-retinoic acid the MTD is also 4 mg/m² weekly x 3 with one week of rest. For 
hematologic indications the MTD is 6 mg/m² weekly x 3 and one week rest.  
 
Approximately 40% variability in clearance of entinostat was noted. When clearance was 
adjusted for body surface area, the inter-patient variability was similar. A linear regression 
analysis on factors that may contribute to this variability, such as ideal body weight, lean body 
mass, body weight, and body mass index, were not significant covariates. As a result of this 
analysis, fixed dosing is considered to be as accurate as dosing based on body surface area17. 
Therefore, based on the clinical experience with entinostat, a dose of 3 and 5 mg given every 2 
weeks has been selected as the regimen for this study. Combination studies with flat every 2 
weeks dose of entinostat are currently conducted in breast and lung cancer. Antitumor response 
has been reported in patients with melanoma treated with 3 mg every 2 weeks dose.  
 
Our preclinical data suggest that the immunomodulatory effect of entinostat is observed 7-14 
days after administration. The every 2 week schedule will allow the pharmacodynamic 
assessment of entinostat as single agent and eventually prime the immune response to high dose 
aldesleukin administration.  
 
3. PATIENT SELECTION 
 
3.1 
Eligibility Criteria 
 
3.1.1 Patients must have pathological diagnosis of renal cell carcinoma that is 
metastatic or surgically unresectable. The histology must be clear cell carcinoma 
or predominant clear cell carcinoma. 
  
3.1.2 Patients may have received up to two prior therapies including VEGF, mTOR and 
PD-1/PDL1 inhibitors.Prior palliative radiation to metastatic lesion(s) is 
permitted, provided there is at least one measurable and/or evaluable lesion(s) that 
has not been irradiated. 
 
3.1.3 Patients must have measurable or evaluable disease. 
 

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3.1.4 Age > 18 years. 
 
3.1.5 ECOG performance status 0 (see Appendix B). 
 
3.1.6 Life expectancy of greater than 6 months. 
 
3.1.7 Patients must have normal organ and marrow function as defined below: 
• Hemoglobin 
 ≥12 g/dL 
• Leukocytes 
 >3,000/mm3 
• Absolute neutrophil count 
 >1,500/mm3 
• Platelets 
 
 >100,000/mm3 
• Total bilirubin 
<1.5 x laboratory upper limit of  normal 
• AST(SGOT)/ALT(SGPT) 
2.5 x laboratory upper limit of normal 
• Creatinine  
≤1.5 x laboratory upper limit of normal    
                                                                  or Calculated Creatinine clearance of  
                                                                 ≥50ml/min (please use institutional  
                                                                  formula) 
• LDH 
 
 WNL 
• Corrected calcium 
 ≤10 mg/dL 
• PT/INR 
 
 ≤1.5 
• Urine protein 
 <1+; if ≥1+, 24 hour urine protein should   
                                                                               be obtained and should be <1000 mg 
 
3.1.8 Pulmonary: FEV1 > 2.0 liters or > 75% of predicted for height and age. (PFTs are 
required for patients over 50 or with significant pulmonary or smoking history). 
 
3.1.9 Cardiac: No evidence of congestive heart failure, symptoms of coronary artery 
disease, myocardial infarction less than 6 months prior to entry, serious cardiac 
arrhythmias, or unstable angina. Patients who are over 40 or have had previous 
myocardial infarction greater than 6 months prior to entry will be required to have 
a negative or low probability cardiac stress test for cardiac ischemia. 
 
3.1.10 CNS: No history of cerebrovascular accident or transient ischemic attacks. 
 
3.1.11 The effects of entinostat on the developing human fetus at the recommended 
therapeutic dose are unknown. For this reason women of child-bearing potential 
must agree to use adequate contraception (hormonal or barrier method of birth 
control; abstinence) prior to study entry and for the duration of study 
participation. Should a woman become pregnant or suspect she is pregnant while 
participating in this study, she should inform her treating physician immediately. 
Men with female partners of child bearing potential must also agree to use 
adequate contraception. 
 

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3.1.12 Ability to understand and the willingness to sign a written informed consent 
document. 
3.2 
Exclusion Criteria 
 
3.2.1 Patients who have received more than two prior therapies. 
 
 
3.2.2 Concurrent use of valproic acid is not allowed. 
 
3.2.3 Patients may not be receiving any other investigational agents. 
 
3.2.4 Patients with untreated CNS metastases. Patients should have a head CT/MRI 
within 28 days prior to treatment initiation. Patients with previously 
excised/gamma knifed solitary or oligometastases and controlled disease are 
eligible. 
 
3.2.5 Any medical condition that would preclude adequate evaluation of the safety and 
 
toxicity of the study combination.  
 
3.2.6 Uncontrolled intercurrent illness including, but not limited to, ongoing or active 
infection, symptomatic congestive heart failure (New York Association Class II, 
III, or IV), angina pectoris requiring nitrate therapy, recent myocardial infarction 
(< the last 6 months), cardiac arrhythmia, history of CVA within 6 months, 
hypertension (defined as blood pressure of >160 mmHg systolic and/or >90 
mmHg diastolic on medication) history of peripheral vascular disease, or 
psychiatric illness/social situations that would limit compliance with study 
requirements.  
 
3.2.7 Patients with a history of allergy to entinostat or other medications that have a 
benzamide structure (i.e. tiapride, remoxipride, and clebopride). 
 
3.2.8 Pregnant women are excluded from this study because entinostat is a HDAC 
inhibitor agent with the potential for teratogenic or abortifacient effects. Because 
there is an unknown but potential risk for adverse events in nursing infants 
secondary to treatment of the mother with entinostat, breastfeeding should be 
discontinued if the mother is treated with entinostat. 
 
3.2.9 HIV-positive patients receiving combination antiretroviral therapy are ineligible 
because of the potential for pharmacokinetic interactions with entinostat. In 
addition, these patients are at increased risk of lethal infections when treated with 
marrow-suppressive therapy. Appropriate studies will be undertaken in patients 
receiving combination anti-retroviral therapy when indicated. 
 
3.2.10 Serious or non-healing wound, ulcer or bone fracture. 
 
3.2.11 Major surgical procedure, open biopsy, or significant traumatic injury within 28 

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days prior to day 1 therapy. 
 
3.2.12 Anticipation of need for major surgical procedures during the course of the study 
 
3.2.13 Left ventricular ejection function <45%. 
 
3.3 
Inclusion of Women and Minorities 
 
 
Both men and women of all races and ethnic groups are eligible for this trial. 
 
 
4. TREATMENT PLAN 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment will be administered on an inpatient/outpatient basis. Patients will be admitted to the 
hospital unit with intensive care capabilities for the administration of high dose aldesleukin. 
Reported adverse events and potential risks are described in Section 5. Appropriate dose 
modifications for entinostat and aldesleukin are described in Section 5. No investigational or 
commercial agents or therapies other than those described below may be administered with the 
intent to treat the patient’s malignancy. 
 
Day
Entinostat
-
Aldesleukin
Treatment Schema
8
9
10 11 12 13 14
1
2
3
4
5
6
7
-2
-1
-4
-3
-6
-5
- 7
15 16 17 18
19
- 8
- 10 - 9
- 12-11
-14 -13
Aldesleukin
Day
Entinostat
-
Aldesleukin
Treatment Schema
8
9
10 11 12 13 14
1
2
3
4
5
6
7
-2
-1
-4
-3
-6
-5
- 7
15 16 17 18
19
- 8
- 10 - 9
- 12-11
-14 -13
Aldesleukin
Cycle 1
Cycle 2
…
Treatment
Days
- 14
1
15
29
43
57
71
85
99
...
X
X
X
X
X
X
...
Entinostat
Aldesleukin
X
X
X
X
92
X
X
PET/CT scan 
(week 5)
PET/CT scan
(baseline)
Tumor response assessment 
X

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The case report form will capture the concurrent use of all other drugs, over-the-counter 
medications, or alternative therapies. 
 
4.1 
Entinostat / aldesleukin administration and definition of Dose-Limiting Toxicity 
(DLT) 
Patients with metastatic renal cell carcinoma and no prior therapies will be eligible for treatment. 
One cycle of treatment (84 days) will consist of 2 courses of high dose aldesleukin 600,000 
units/kg administered IV every 8 hrs. on day 1-5 and day 15-19 (± 7 days) (maximum 28 doses), 
and entinostat orally (1-2 hrs. prior to aldesleukin infusion) given every 2 weeks starting on day-
14, continuously. Entinostat is to be taken on an empty stomach, at least 1 hour before or 2 hours 
after a meal. Tumor response assessment will be performed on week 11 (+/- 7 days). In the event 
of clinical benefit (stable disease or tumor shrinkage) patients will receive a second cycle of 
therapy. 
 
Entinostat will continue every 2 weeks (+/- 7 days dependent on adjustments necessary for 
aldesleukin dosing) until documented disease progression or 8 weeks following documented 
complete response. Patients with evidence of tumor shrinkage may receive up to 3 cycles of high 
dose aldesleukin therapy. Cycle 2 will start on or within 2 weeks following day 85 (or last day of 
cycle dependent on dosing adjustment) as in cycle 1. Cycle 3 can start within 2 weeks after 
completion of cycle 2. Patients with stable disease by RECIST V.1.0 criteria, but without 
evidence of tumor shrinkage after two cycles will receive only entinostat until disease 
progression is documented. 
 
In order to assess the effect of entinostat versus the combination on the proposed correlative 
pharmacodynamic parameters, initial treatment will be with entinostat monotherapy, followed by 
combination with high dose aldesleukin. The Phase I starting dose level of entinostat will be 3 
mg PO every 2 weeks. The first dose level will have a minimum of 3 patients treated unless the 
first 2 patients experience dose-limiting toxicity(s) (DLT) before 3rd patient is enrolled. DLTs are 
defined as extended grade 4 toxicity (duration of one week or more) during the first 45 days of 
treatment. If DLTs occur in 1 patient treated at the starting dose level, a minimum of 3 or 4 
patients will be treated at this dose level. If DLT occurs in more than 1 patient in the first 6 
patients, patients will be treated at a lower dose level (entinostat 1 mg PO every 2 weeks, dose 
level 0) and DLT will be evaluated similarly. If DLTs occur in ≤ 1/6 patients, dose level 1 will 
be considered the recommended Phase II dose. If no DLTs occur at the starting dose level 1, 3 or 
4 patients will be treated at the next dose level (level 2) with 5 mg PO every 2 weeks (wait 45 
days for DLT assessment on first 2 patients before 3rd patient is enrolled). If no DLTs occur at 
dose level 2, this dose level will be recommended for the phase II portion of the study. If DLTs 
occur in ≥ 1 patient, another 3 patients will be treated at this dose level. If DLT occurs in ≥ 2 
patients in the first 6 patients, dose level 1 will be the recommended for the Phase II. If DLTs 
occur in ≤ 1/6 patients, dose level 2 will be considered the recommended Phase II dose. Patients 
who experience extended grade 4 toxicity (duration of one week or more) and recover to ≤ Grade 
1 (or to pretreatment baseline level toxicity) may continue treatment at the next entinostat lower 
level. Once the entinostat has been dose reduced, patients must remain at the reduced dose for 
the rest of the study. Patients may have their dose de-escalated twice or up to dose level 0 before 
being removed from the treatment. Patients will be allowed to remain on the therapy provided 
that they are tolerating the treatment and do not develop progressive disease. No dose de-

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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escalation for aldesleukin will be performed. 
 
Dosing levels 
Dose Level 
Entinostat (mg, every 2 weeks, PO) 
Aldesleukin (IU/kg, every 8hrs, IV) 
0 
1 
600,000 
1 
3 
600,000 
2 
5 
600,000 
 
Patients will receive only entinostat during the “maintenance” (between cycles) period. During 
Phase II, patients receiving 5 mg entinostat during the “maintenance” period will be given the 
option to dose escalate entinostat to 10 mg PO if tolerated except two weeks before and two 
weeks after administration of aldesleukin.  
 
The recommended dose of 5 mg PO every 2 weeks was determined in the Phase I portion of the 
study. This cohort will be expanded in the phase II portion so that 36 new patients will be treated 
at this dose level. The evaluations for the phase II component of the study will be identical to the 
phase I portion.  
 
Compliance will be assessed via patient diaries Also, pill containers will be returned to the study 
doctor or nurse at the end of each cycle. 
 
4.2 
Duration of Therapy 
Patients may continue on therapy until one of the following criteria applies: 
 
• Disease progression, 
• Unacceptable toxicity. 
• Patient decides to withdraw from the study. 
• General or specific changes in the patient’s condition that renders the patient 
unacceptable for further treatment in the judgment of the investigator. 
• The study is closed. 
 
4.3 
Duration of Follow Up 
 
Follow up safety evaluations will occur 30 days (+/- 4 days) after last dose of study drug or until 
resolution of any drug related toxicity (telephone contact is acceptable). Follow up will include 
review of concomitant medications and adverse events. 
 
Patients (both Phase I and Phase II) will be followed up after completion of therapy every three 
months for survival assessment (telephone contact is acceptable) or until completion of primary 
endpoint/ termination of study. 
 
5. DOSING DELAYS/DOSE MODIFICATIONS 
 

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The CTEP Active Version of the NCI Common Terminology Criteria for Adverse Events 
(CTCAE) [http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm will be 
used to classify toxic effects of therapy. Patients with life-threatening or persistent, severe toxic 
reactions to aldesleukin will receive no further treatment with this agent. Specific toxicities, 
which require special consideration, are discussed below. Previously unknown or severe 
toxicities will be reported to the PI as Adverse Drug Reactions. 
 
5.1 
Entinostat Treatment Modification for Adverse Events 
 
Patients who experience any Grade 3/4 toxicity related to entinostat during the period off 
aldesleukin should stop the offending drug up to one dose. If the patients recover to < 
Grade 1 (or to pretreatment baseline level of toxicity), they may continue treatment with 
entinostat at the next lower dose. Once the dose has been reduced, patients must remain at the 
reduced dose for the remainder of the study. Patients may have their dose reduced down to the 
lowest dose level 0 (1mg). If the patient misses or vomits a dose of entinostat he/she will skip 
that dose and notify the research nurse. If a patient has to miss more than one dose of entinostat 
due to grade 3/4 toxicity during the period off aldesleukin he/she will discontinue entinostat.  
 
5.2 
Adverse Events with entinostat 
 
Risks to patients receiving entinostat have been minimized with careful monitoring by trained 
and experienced investigators and evaluation of laboratory safety parameters at appropriate 
intervals. The following points should be considered for the prevention or minimization of 
potential adverse events and other risks related to entinostat:  
 
• Nausea and vomiting: Antiemetics may be administered to prevent or control nausea or 
vomiting.  
 
 • Diarrhea: Antidiarrheal agent(s) may be used in the event of diarrhea. 
  
 • Neutropenia: The patient should be closely followed for fever, focal signs of infection, and 
neutrophil nadir. Standard growth factor support may be considered unless excluded in the 
individual clinical protocols.  
  
 • Anemia: RBC transfusion support may be given for anemia, particularly when hemoglobin 
falls below 8 g/dL or as instructed in the individual clinical protocols.  
  
 • Thrombocytopenia: If bleeding develops or invasive procedures are planned, platelet 
transfusions may be administered in accordance with standard practice or as instructed in the 
individual clinical protocol.  
  
 • Hypophosphatemia and other electrolyte abnormalities: Careful monitoring of electrolytes 
during treatment with entinostat is recommended and electrolyte supplementation may be 
warranted. In clinical studies, abnormalities in most patients were successfully managed with 
supplementation, allowing continued treatment without interruption. 
  

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 • Renal abnormalities: Based on mild increases in creatinine seen in a few patients and  
  results of nonclinical studies, monitoring of renal function is recommended during  
  treatment with entinostat. If clinically significant changes in renal function occur,  
  modification of entinostat dose should be considered.  
  
Entinostat may cause fatigue or malaise; advise patient to exercise caution while driving a 
vehicle or operating machinery.  
 
Administration of entinostat is contraindicated in patients with a history of allergy to entinostat 
or other medications that have a benzamide structure (e.g., tiapride, remoxipride, clebopride). 
 
Careful monitoring of patients for signs of infection or reactivation of past infections is 
recommended, as reactivation of infection has been reported in patients treated with entinostat, in 
some cases without evidence of neutropenia. The clinical significance of this finding and the 
potential association with entinostat is unknown. 
 
 
5.3 
Aldesleukin Treatment Modification for Adverse Events 
 
Modification of the treatment protocol will occur by withholding doses of aldesleukin rather than 
continuing therapy at a reduced dose (see Appendix C). Missed doses of aldesleukin will not be 
made up. Dose of aldesleukin will be withheld for: 
 
5.31 
Hypotension refractory to fluids and pressors or requiring unacceptably high 
 
pressor doses 
 
5.3.2 Anuria for>24 hours and unresponsive to fluid replacement and low-dose 
 
dopamine 
 
5.3.3 Respiratory distress requiring oxygen>4 liters to maintain O2 saturation>95%. 
 
5.3.4 Confusion (mental status changes can progress to paranoia despite discontinuation 
 
of aldesleukin; it is imperative that the aldesleukin be stopped at any sign of 
 
persistent confusion or disorientation). 
 
5.3.5 Sustained ventricular tachycardia or any sign or symptom of myocardial ischemia 
 
or myocarditis. Patients experiencing sustained ventricular tachycardia or 
 
myocardial ischemia should not receive further treatment with aldesleukin 
 
5.3.6  Metabolic acidosis with HCO3<18, despite attempts to correct with IV HCO3. 
 
5.3.7  Atrial fibrillation or myocarditis. 
 
5.3.8  Documented systemic infection. 
 
5.3.9  Any other serious toxicity that is not controlled at time of next dose. 

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5.4 
Specific aldesleukin Toxicity Management 
 
Several treatment-related toxicities have been uniquely associated with the administration of 
high-dose aldesleukin. The frequency of Grade 3 and Grade 4 toxicities in Cycle 1 ranges, for 
example, from 56.8% hypotension to 14.7% neurological, and 13.7% hematological, 
neurological, pulmonary, renal/electrolytes. Antihypertensive medication should be stopped prior 
to aldesleukin therapy. 
 
Recommendations for management of the more significant toxicities typically seen with high-
dose bolus aldesleukin are as follows (the following are only optional guidelines for toxicity 
management): 
 
5.4.1 Fluid Replacement. 
Excessive fluid replacement will increase the patient’s likelihood of developing 
pulmonary edema. It is suggested that when intravenous access has been 
established; begin administration of normal saline at 75cc/hour IV. Fluid 
replacement should be given to allow patients to gain approximately 1kg/day. 
Once patients have gained greater than 5% of baseline weight, additional fluid 
boluses should not be given to maintain blood pressure. Furosemide should not be 
given unless symptomatic fluid retention develops and blood pressure is adequate. 
 
5.4.2 Hypotension 
Administration of high dose aldesleukin leads to decreased peripheral vascular 
resistance and consequent hypotension. In order to manage this toxicity, the 
physician may utilize the following guidelines: 
 
1. Monitor patients in a setting capable of providing ICU level care. 
  
 
2. Prior to starting aldesleukin therapy determine a minimum tolerated blood 
pressure (MTBP). For patients under age 40 and with no prior history of 
ischemic or valvular heart disease, the MTBP can be a systolic blood pressure 
(SBP) of 80 mmHg, while the MTBP for all other patients should be a SBP of 
85-90 mmHg based on perceived risk of cardiac toxicity. 
 
3. When a patients systolic BP falls below the MTBP, suggested therapy may 
 involve (in the following order):  
a. Begin fluid boluses (250cc normal saline IV over 15 min. may 
                  repeat x 2) until SBP is > MTBP. 
 
             b. Should fluid boluses fail make the SBP>MTBP, phenylephrine should  
                  also be instituted (0.1-2 mcg/kg/min) to maintain SBP. 
 
      c. If blood pressure cannot be maintained on phenylephrine alone, or if  
          unacceptable tachycardia develops, treatment with dopamine  
          (1-6 mcg/kg/min) should be added to sustain blood pressure. 

Entinostat and Aldesleukin 
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5.4.3 Management of Arrhythmias/Myocarditis.  
If significant arrhythmias occur at any stage in the patient's treatment (whether on 
pressor agents or not), the possibility of myocardial ischemia/infarction must be 
excluded by both EKG and cardiac enzyme assessment. Patients who develop 
atrial fibrillation should have aldesleukin doses held. Therapy may resume when 
the patient converts to normal sinus rhythm and is hemodynamically stable. If a 
significant supraventricular arrhythmia occurs while a patient is on dopamine 
therapy; phenylephrine should also be substituted for dopamine as initial blood 
pressure support. Patients experiencing sustained ventricular tachycardia or 
documented myocardial ischemic episodes during therapy should not receive 
further treatment with aldesleukin. Patients with myocarditis may resume 
treatment in subsequent courses if CPK returns to normal. A cardiac MUGA 
stress test documenting normal cardiac function should be performed prior to 
restarting therapy. 
 
5.4.4 Management of Neurotoxicity  
Doses are held rather than reduced for neurotoxicity. If Grade 4 neuro-cortical 
toxicity is encountered and is not reversible within 48 hours, no further treatment 
should be given and the patient should be removed from treatment. If Grade 4 
toxicity is reversible to Grade 1 within 48 hours, future treatment may be 
considered (in subsequent courses) if the patients shows any evidence of tumor 
regression. 
 
5.4.5  Metabolic Acidosis 
In the course of aldesleukin therapy, when a patient’s HCO3 falls to below 20, 
NaHCO3 should be added to the maintenance IV infusion. Should the HCO3 
level fall below 18, aldesleukin therapy should be held, and bolus infusions of 
NaHCO3 should be instituted. Aldesleukin therapy may resume if repeat HCO3 > 
18. 
 
5.4.6 Other Toxicities 
Increases in the serum creatinine to 2.0-3.5 mg/dl and total bilirubin to 3.0-10.0 
mg/dl are common and reversible upon cessation of treatment. Doses of 
aldesleukin have not generally been withheld for renal and hepatic dysfunction 
alone. Although toxicity may become severe, recovery usually occurs following 
cessation of aldesleukin and vigorous supportive care is warranted. 
 
5.5 
Management of Grade 4 Toxicity 
 
Patients with Grade 4 (Life Threatening) toxicity may be treated with dexamethasone 4 mg qid 
until side effects improve to an acceptable level. In the clinical trials with aldesleukin, 
dexamethasone has been used to treat patients with pulmonary edema requiring assisted 
ventilation, although many patients may be managed successfully without steroids. 
 

Entinostat and Aldesleukin 
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Protocol Version Date  5/29/19 
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5.6 
Toxicity Criteria for Discontinuing Treatment 
 
Patients will not be considered for further therapy if the following toxicities are encountered: 
  
  
5.6.1 Pulmonary toxicity requiring endotracheal intubation 
  
 
5.6.2  Renal dysfunction requiring dialysis 
 
5.6.3  Grade 4 cardiac dysrhythmia or Grade 2 or 3 dysrhythmia not easily controlled 
with medical management 
 
5.6.4 Myocardial ischemia (Grade 3 or 4) or infarction or symptomatic myocarditis 
(note: asymptomatic CPK or CPK-MB band elevations without EKG changes are 
not a contraindication to further treatment). 
 
 
5.6.5  Coma 
 
 
5.6.6 Life-threatening sepsis 
 
 
5.6.7 Pericardial tamponade 
 
 
5.6.8 Bowel ischemia or perforation 
 
5.6.9 Grade 4 hypertension or hemorrhage, symptomatic grade 4 venous 
thromboembolic event, nephritic syndrome 
 
5.6.10  Any grade arterial thromboembolic event 
 
5.6.11 Inability of subject to comply with study requirements 
 
5.6.12 Any other severe or life-threatening toxicity which, in the opinion of the 
investigator, would preclude further treatment with these agents, or has not 
resolved to baseline 8 weeks after treatment. 
 
6. PHARMACEUTICAL INFORMATION 
 
6.1 
Entinostat (NSC 706995) 
 
Chemical name: 3-Pyridylmethyl N-{4-[(2-aminophenyl)carbamoyl]benzyl}carbamate  
 
Other names:  
MS-27-275, MS-275, SNDX-275 
 
Classification:  
Histone deacetylase inhibitor (HDACi)  
 

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Molecular formula:  
C21H20N4O3 
 
 
 
 
M.W.: 376.41 
 
Mode of Action: 
Histone deacetylases (HDACs) are a family of enzymes that regulate 
chromatin remodeling and gene transcription via the dynamic process of acetylation and 
deacetylation of core histones.  Entinostat inhibits histone deacetylases, changes chromatin 
configuration, and induces differentiation and apoptosis of cancer cells through an 
epigenetic mechanism. 
 
How Supplied:  
Entinostat is supplied by the Syndax Pharmaceuticals, Inc. and 
distributed by DCTD, NCI as 1 mg (pink to light red, in bottles of 40), or 5 mg (yellow, in 
bottles of 40) film-coated tablets (round-biconvex). Each tablet also contains mannitol, 
sodium starch glycolate, hydroxypropyl cellulose, potassium bicarbonate, and magnesium 
stearate.  The film coating consists of hypromellose, talc, titanium dioxide, and ferric oxide 
pigments (red and yellow) as colorants.   
 
Matching placebo for entinostat has the same appearance as the corresponding active 
tablets and contains the same inactive ingredients and film coating. 
 
Storage: Store the bottles at controlled room temperature (15-25oC), and protect from light.  
Entinostat is not to be exposed to extremes of temperature (greater than 30ºC or less than 
5ºC). 
 
Stability: 
Shelf life stability studies of the intact bottles are on-going. 
 
Route of Administration: 
Oral, on an empty stomach, at least 1 hour before or 2 
hours after a meal.  Entinostat tablets should not be split, crushed, or chewed. 
 
6.1.1 Availability 
Entinostat is an investigational agent supplied to investigators by the Division of Cancer  
 
Treatment and Diagnosis (DCTD), NCI. 
 
Entinostat is provided to the NCI under a Collaborative Agreement between Syndax 
Pharmaceuticals, Inc.. and the DCTD, NCI. 
 
6.1.2 Agent Ordering 
 
NCI-supplied agents may be requested by the Principal Investigator (or their authorized 
designees) at each participating institution. Pharmaceutical Management Branch (PMB) 
policy requires that the agent be shipped directly to the institution where the patient is to 

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Protocol Version Date  5/29/19 
page 24 of 78 
 
be treated. PMB does not permit the transfer of agents between institutions (unless prior 
approval from PMB is obtained). The CTEP-assigned protocol number must be used for 
ordering all CTEP-supplied investigational agents. The responsible investigator at each 
participating institution must be registered with CTEP, DCTD through an annual 
submission of FDA Form 1572 (Statement of Investigator), Curriculum Vitae, 
Supplemental Investigator Data Form (IDF), and Financial Disclosure Form (FDF). If 
there are several participating investigators at one institution, CTEP-supplied 
investigational agents for the study should be ordered under the name of one lead 
investigator at that institution. 
 
 
Active CTEP-registered investigators and investigator-designated shipping designees and 
ordering designees can submit agent requests through the PMB Online Agent Order 
Processing (OAOP) application (https://eapps-ctep.nci.nih.gov/OAOP/pages/login.jspx). 
Access to OAOP requires the establishment of a CTEP Identity and Access Management 
(IAM) account (https://eapps-ctep.nci.nih.gov/iam/) and the maintenance of an “active” 
account status and a “current” password. For questions about drug orders, transfers, 
returns, or accountability, call (240) 276-6575 Monday through Friday between 8:30 am 
and 4:30 pm (ET) or email PMBAfterHours@mail.nih.gov anytime. 
 
6.1.3  Agent Inventory Records 
The Investigator, or a responsible party designated by the investigator, must maintain a 
careful record of the inventory and disposition of all agents received from DCTD using 
the NCI Oral Drug Accountability Record Form (Oral DARF). (See the NCI 
Investigator’s Handbook for Procedures for Drug Accountability and Storage.) 
 
6.1.4 
 Comprehensive Adverse Events and Potential Risks list (CAEPR) 
 for Entinostat (SNDX-275, MS-275, NSC 706995) 
 
The Comprehensive Adverse Event and Potential Risks list (CAEPR) provides a single list of 
reported and/or potential adverse events (AE) associated with an agent using a uniform 
presentation of events by body system. In addition to the comprehensive list, a subset, the 
Specific Protocol Exceptions to Expedited Reporting (SPEER), appears in a separate column and 
is identified with bold and italicized text. This subset of AEs (SPEER) is a list of events that are 
protocol specific exceptions to expedited reporting to NCI via CTEP-AERS (except as noted 
below). Refer to the 'CTEP, NCI Guidelines: Adverse Event Reporting Requirements' 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/docs/aeguidelines.pdf for further 
clarification. Frequency is provided based on 215 patients. Below is the CAEPR for entinostat 
(SNDX-275, MS-275). 
 
NOTE: Report AEs on the SPEER ONLY IF they exceed the grade noted in parentheses next to 
the AE in the SPEER. If this CAEPR is part of a combination protocol using multiple 
investigational agents and has an AE listed on different SPEERs, use the lower of the grades to 
determine if expedited reporting is required. 
 
 

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Protocol Version Date  5/29/19 
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Version 2.5, September 10, 20181 
 
 Adverse Events with Possible  
 Relationship to MS-275 (SNDX-275, entinostat) 
 (CTCAE 5.0 Term) 
[n= 221] 
 
 
 Specific Protocol Exceptions to 
Expedited Reporting (SPEER) 
 
Likely (>20%) 
Less Likely (<=20%) 
Rare but Serious (<3%)  
 
BLOOD AND LYMPHATIC SYSTEM DISORDERS 
  
Anemia 
 
 
 Anemia (Gr 3) 
GASTROINTESTINAL DISORDERS 
  
 
Abdominal pain 
 
 Abdominal pain (Gr 2) 
 
Constipation 
 
 Constipation (Gr 2) 
 
Diarrhea 
 
 Diarrhea (Gr 3) 
 
Dyspepsia 
 
 Dyspepsia (Gr 2) 
Nausea 
 
 
 Nausea (Gr 3) 
Vomiting 
 
 
 Vomiting (Gr 3) 
GENERAL DISORDERS AND ADMINISTRATION SITE CONDITIONS 
  
 
Edema limbs 
 
 Edema limbs (Gr 2) 
Fatigue 
 
 
 Fatigue (Gr 3) 
 
Fever 
 
 Fever (Gr 2) 
INFECTIONS AND INFESTATIONS 
  
 
Infection2 
 
 Infection2 (Gr 3) 
INVESTIGATIONS 
  
 
Alkaline phosphatase increased 
 
 Alkaline phosphatase increased  
(Gr 2) 
 
Lymphocyte count decreased 
 
 Lymphocyte count decreased (Gr 4) 
Neutrophil count decreased 
 
 
 Neutrophil count decreased (Gr 4) 
Platelet count decreased 
 
 
 Platelet count decreased (Gr 4) 
 
White blood cell decreased 
 
 White blood cell decreased (Gr 3) 
METABOLISM AND NUTRITION DISORDERS 
  
Anorexia 
 
 
 Anorexia (Gr 3) 
 
Dehydration 
 
 Dehydration (Gr 2) 

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Protocol Version Date  5/29/19 
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 Adverse Events with Possible  
 Relationship to MS-275 (SNDX-275, entinostat) 
 (CTCAE 5.0 Term) 
[n= 221] 
 
 
 Specific Protocol Exceptions to 
Expedited Reporting (SPEER) 
 
Likely (>20%) 
Less Likely (<=20%) 
Rare but Serious (<3%)  
 
 
Hyperglycemia 
 
 Hyperglycemia (Gr 2) 
Hypoalbuminemia 
 
 
 Hypoalbuminemia (Gr 2) 
 
Hypocalcemia 
 
 Hypocalcemia (Gr 2) 
 
Hypokalemia 
 
 Hypokalemia (Gr 2) 
 
Hyponatremia 
 
 Hyponatremia (Gr 3) 
Hypophosphatemia 
 
 
 Hypophosphatemia (Gr 3) 
MUSCULOSKELETAL AND CONNECTIVE TISSUE DISORDERS 
  
 
Myalgia 
 
 Myalgia (Gr 2) 
NERVOUS SYSTEM DISORDERS 
  
 
Dysgeusia 
 
 Dysgeusia (Gr 2) 
Headache 
 
 
 Headache (Gr 2) 
RESPIRATORY, THORACIC AND MEDIASTINAL DISORDERS 
  
 
Cough 
 
 Cough (Gr 2) 
 
Dyspnea 
 
 Dyspnea (Gr 3) 
SKIN AND SUBCUTANEOUS TISSUE DISORDERS 
  
 
 
Erythema multiforme 
  
 
1This table will be updated as the toxicity profile of the agent is revised.  Updates will be distributed to all Principal 
Investigators 
at 
the 
time 
of 
revision. 
 
The 
current 
version 
can 
be 
obtained 
by 
contacting 
PIO@CTEP.NCI.NIH.GOV. Your name, the name of the investigator, the protocol and the agent should be included 
in the e-mail. 
 
2Infection includes all 75 sites of infection under the INFECTIONS AND INFESTATION SOC. 
 
3Gastrointestinal hemorrhage includes Anal hemorrhage, Cecal hemorrhage, Colonic hemorrhage, Duodenal 
hemorrhage, Esophageal hemorrhage, Esophageal varices hemorrhage, Gastric hemorrhage, Hemorrhoidal 
hemorrhage, Ileal hemorrhage, Intra-abdominal hemorrhage, Jejunal hemorrhage, Lower gastrointestinal 
hemorrhage, Oral hemorrhage, Pancreatic hemorrhage, Rectal hemorrhage, Retroperitoneal hemorrhage, and Upper 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 27 of 78 
 
gastrointestinal hemorrhage under the GASTROINTESTINAL DISORDERS SOC. 
 
 
 
Adverse events reported on MS-275 (SNDX-275, entinostat) trials, but for which there is insufficient evidence 
to suggest that there was a reasonable possibility that MS-275 (SNDX-275, entinostat) caused the adverse 
event: 
 
BLOOD AND LYMPHATIC SYSTEM DISORDERS - Febrile neutropenia; Hemolysis; Leukocytosis 
CARDIAC DISORDERS - Atrial fibrillation; Atrioventricular block complete; Cardiac disorders - Other (transient 
right-side heart failure with worsening tricuspid regurgitation); Chest pain - cardiac; Conduction disorder; Heart 
failure; Left ventricular systolic dysfunction; Palpitations; Pericardial effusion; Pericarditis; Sinus tachycardia; 
Supraventricular tachycardia; Ventricular fibrillation 
EAR AND LABYRINTH DISORDERS - Hearing impaired 
EYE DISORDERS - Blurred vision 
GASTROINTESTINAL DISORDERS - Anal mucositis; Colitis; Dysphagia; Enterocolitis; Esophageal pain; 
Esophagitis; Flatulence; Gastrointestinal disorders - Other (hyperdefecation); Gastrointestinal hemorrhage3; 
Hemorrhoids; Mucositis oral; Pancreatitis; Periodontal disease; Rectal mucositis; Rectal pain; Small intestinal 
mucositis; Typhlitis; Visceral arterial ischemia 
GENERAL DISORDERS AND ADMINISTRATION SITE CONDITIONS - Chills; Edema face; Generalized 
edema; Injection site reaction; Multi-organ failure; Non-cardiac chest pain; Pain 
IMMUNE SYSTEM DISORDERS - Allergic reaction; Anaphylaxis; Autoimmune disorder 
INJURY, POISONING AND PROCEDURAL COMPLICATIONS - Bruising 
INVESTIGATIONS - Activated partial thromboplastin time prolonged; Alanine aminotransferase increased; 
Aspartate aminotransferase increased; Blood bilirubin increased; CPK increased; Creatinine increased; GGT 
increased; INR increased; Investigations - Other (coagulopathy); Investigations - Other (vitamin D deficiency); 
Lipase increased; Serum amylase increased; Weight loss 
METABOLISM 
AND 
NUTRITION 
DISORDERS 
- 
Acidosis; 
Hypercalcemia; 
Hyperkalemia; 
Hypermagnesemia; Hypernatremia; Hypertriglyceridemia; Hyperuricemia; Hypoglycemia; Hypomagnesemia; 
Tumor lysis syndrome 
MUSCULOSKELETAL AND CONNECTIVE TISSUE DISORDERS - Arthralgia; Back pain; Bone pain; Chest 
wall pain; Generalized muscle weakness; Muscle cramp; Musculoskeletal and connective tissue disorder - Other 
(thorax pain); Myositis; Pain in extremity 
NEOPLASMS BENIGN, MALIGNANT AND UNSPECIFIED (INCL CYSTS AND POLYPS) - Tumor pain 
NERVOUS SYSTEM DISORDERS - Ataxia; Depressed level of consciousness; Dizziness; Dysphasia; 

Entinostat and Aldesleukin 
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Protocol Version Date  5/29/19 
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Intracranial hemorrhage; Neuralgia; Olfactory nerve disorder; Peripheral motor neuropathy; Peripheral sensory 
neuropathy; Seizure; Syncope; Tremor 
PSYCHIATRIC DISORDERS - Anxiety; Confusion; Depression; Insomnia; Libido decreased 
RENAL AND URINARY DISORDERS - Acute kidney injury; Proteinuria; Renal and urinary disorders - Other 
(bladder distension); Renal calculi; Renal hemorrhage; Urinary frequency; Urinary retention 
RESPIRATORY, THORACIC AND MEDIASTINAL DISORDERS - Allergic rhinitis; Atelectasis; Epistaxis; 
Hypoxia; Laryngeal mucositis; Pharyngeal mucositis; Pleural effusion; Pleuritic pain; Pulmonary edema; 
Respiratory failure; Tracheal mucositis 
SKIN AND SUBCUTANEOUS TISSUE DISORDERS - Alopecia; Hyperhidrosis; Nail loss; Photosensitivity; 
Pruritus; Purpura; Rash maculo-papular; Skin and subcutaneous tissue disorders - Other (hyperkeratotic 
lesions/squamous cell carcinoma); Urticaria 
SURGICAL AND MEDICAL PROCEDURES - Surgical and medical procedures - Other (packed RBC 
transfusion) 
VASCULAR DISORDERS - Flushing; Hypertension; Hypotension; Thromboembolic event 
 
Note: MS-275 (SNDX-275, entinostat) in combination with other agents could cause an exacerbation of any adverse 
event currently known to be caused by the other agent, or the combination may result in events never previously 
associated with either agent. 
 
 
6.1.5 Potential Drug Interactions  
 
Metabolism: Data from in vitro metabolism experiments in human tissues demonstrated that 
entinostat is not metabolized by CYP enzymes (Acharya 2006), but UGT 1A4 did metabolize 
entinostat to its M2 glucuronide metabolite. No metabolites could be detected after incubation of 
entinostat in human liver microsomes (Acharya 2006). While inhibition of CYP enzymes 2B6 
and 3A4 was seen, the data show that the degree of the inhibition makes it unlikely that any in 
vivo systemic interactions would occur. Intestinal CYP 3A4 may be inhibited by entinostat. 
However, entinostat did not inhibit any UGT enzymes tested. Entinostat was found to induce 
CYP 1A2, CYP 2C6, and CYP 2B8 as well as UGT 1A4. Finally, entinostat was found to be a 
substrate for P-gp and BCRP transporters, but did not inhibit either of these transport proteins. 
6.1.6 Patient Care Implications 
Entinostat may cause fatigue or malaise; advise patient to exercise caution while driving a 
vehicle or operating machinery.  
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 29 of 78 
 
Administration of entinostat is contraindicated in patients with a history of allergy to entinostat 
or other medications that have a benzamide structure (eg, tiapride, remoxipride, clebropride).  
Careful monitoring of patients for signs of infection or reactivation of past infections is 
recommended, as reactivation of infection has been reported in patients treated with entinostat, in 
some cases without evidence of neutropenia. The clinical significance of this finding and the 
potential association with entinostat is unknown.  
 
Entinostat must not be used during pregnancy or while breast-feeding. Women and men 
participating in entinostat clinical studies must agree to use acceptable contraceptive methods, as 
indicated in the clinical study protocol, during treatment and for 3 months thereafter. 
 
6.2 
Interleukin-2 (Aldesleukin) 
 
Description: Aldesleukin is recombinant formulation of interleukin-2. Aldesleukin is a non-
glycosylated biosynthetic interleukin-2 (also known as T-cell growth factor), which differs only 
slightly in amino acid sequence from the natural compound. Aldesleukin is commercially 
available. 
 
Mechanism of Action: Aldesleukin's effects are essentially identical to those of endogenous 
interleukin-2. Aldesleukin interacts with the high-affinity interleukin 2 receptor expressed on 
cells of the immune system and stimulates a cytokine cascade involving various interferons, 
interleukins, and tumor necrosis factors. Aldesleukin along with other cytokines induce 
proliferation and differentiation of B and T-cells, monocytes, macrophages, and cytotoxic 
lymphocytes which include natural killer (NK) cells, cytotoxic T-cells, tumor-infiltrating 
lymphocytes (TIL), and lymphokine-activated killer (LAK) cells. Aldesleukin's antitumor 
activity is believed to result from activation of cytotoxic lymphocytes; however, the exact 
mechanism is unknown. Whether aldesleukin acts directly or through second messengers is also 
unclear, however, aldesleukin does elevate production of interleukin-1, tumor necrosis factors 
alpha and beta, interferon gamma, and interleukin-6. 
 
Pharmacokinetics: Aldesleukin is administered parenterally. Following a short IV infusion, the 
drug is rapidly distributed to the extravascular and extracellular space as well as to the liver, 
spleen, kidneys, and lungs. Approximately 30% of an administered dose is distributed within the 
plasma. The pharmacokinetics of aldesleukin may be affected by sodium dodecyl sulfate, the 
solubilizing agent in the commercial formulation. In addition, subcutaneous administration with 
albumin produces slightly higher and more prolonged serum concentrations of aldesleukin. 
Following distribution, aldesleukin is cleared from the systemic circulation by the kidneys 
through both glomerular filtration and peritubular extraction. The drug is then metabolized to 
amino acids by renal cells lining the proximal convoluted tubules. Very little drug is excreted 
unchanged in the urine. Following a 5 minute IV infusion, the serum distribution and elimination 
half-life in cancer patients was 13 and 85 minutes, respectively. 
 
Toxicities: Abdominal pain ,alopecia, anemia, angina, anorexia, anuria, arthralgia, ascites, atrial 
fibrillation, azotemia, back pain, chills, cholestasis, conjunctivitis, constipation, diarrhea, 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 30 of 78 
 
drowsiness, dysgeusia, dyspepsia, dyspnea, dysuria, edema, elevated hepatic enzymes, 
eosinophilia, erythema, exfoliative dermatitis, fatigue, fever, GI bleeding, GI perforation, 
glomerulonephritis, hallucinations, headache, hematuria, hepatomegaly, hyperbilirubinemia, 
hyperkalemia, hypoalbuminemia, hypocalcemia , hypokalemia, hypomagnesemia, hyponatremia, 
hypophosphatemia, hypotension, hypovolemia, infection, injection site reaction, interstitial 
nephritis, jaundice, leukocytosis, leukopenia, lymphopenia, malaise, metabolic acidosis, 
metabolic alkalosis, myalgia, myocardial infarction, nausea/vomiting, oliguria, pancreatitis, 
paranoia, petechiae, pleural effusion, premature atrial contractions (PACs), premature ventricular 
contractions (PVCs), proteinuria, pruritus, pulmonary edema, purpura, sinus bradycardia, sinus 
tachycardia, splenomegaly, stomatitis, supraventricular tachycardia (SVT), syncope, tachypnea, 
thrombocytopenia, thrombosis, urticaria, visual impairment, weakness, weight gain, weight loss, 
wheezing, xerosis. 
 
7. CORRELATIVE/SPECIAL STUDIES 
 
7.1 
Pharmacodynamic Studies  
 
PBMCs: 
Relationships between entinostat and aldesleukin exposure and pharmacodynamic effects (e.g. 
histone acetylation in peripheral blood mononuclear cells or PBMCs and changes in T cell subset 
population) will be characterized. 
 
Collect four 8 ml of peripheral blood in CPT tubes for mononuclear cell fraction. Mononuclear 
cell fractions will be obtained in CPT tubes (Beckon Dickinson) sodium citrate using 
manufacture’s guidelines. 
 
Collection time points will be: 
• Prior to treatment on Day -14 and Day-7,  
• Cycle 1 – pre-dose on Day 1 and at 8 hrs. and 24 hrs. after the dose of entinostat,  
• Cycle 1 - pre-dose Day 15  
• Cycle 2 and 3 - pre-dose Day 1  
• Cycle 2 and 3 - pre-dose Day 15  
• At the time of progression. 
 
Samples will be processed, PBMCs isolated and histone acetylation status determined. Blood 
samples will be also analyzed for immune cell sub populations. CD4+, CD4+/Foxp3, CD8+ 
cells, NK cells will be quantitated by FACS analysis. Foxp3 gene expression will be evaluated 
by quantitative RTPCR. Specimens must be shipped at ambient temperature. 
 
Serum for microRNA profiling: 
Collect a serum sample in one 10 ml red top tube to perform microRNA profiling to identify 
predictive biomarkers for response or resistance to therapy and prognostic biomarkers for 
survival. 
 
 
Collection time points will be: 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 31 of 78 
 
• Day-14 pre-dose 
• Cycle 1 – pre-dose on Day 1 
• Cycle 1- pre-dose day 15 
• Cycle 2 and 3 - pre-dose Day 1  
• At the time of progression.  
 
Centrifuge the tube (within 30 min after blood draw) at 1000 g for 10 min at room temperature. 
Remove supernatant (serum) and aliquot ~ 1 ml into 3 microcentrifuge tubes.  Shipped ambient 
same day of collection Monday through Thursday to Dr. Pili’s lab for analysis.  If collected on 
Friday, weekends, or holiday store refrigerated and shipped ambient the next business day.  
 
Sample Shipment 
All specimens should be correctly labeled with patient initials, study-specific subject ID number, 
protocol number, time point, and date and time of collection. NOTE: In the event that baseline 
specimens are available but the patient is not successfully registered to the protocol, do not 
submit the patient's specimens. Specimens must be shipped for overnight delivery Monday 
through Thursday.  
 
Address shipments and any question regarding specimen processing to: 
 
Notification of sample shipment should be sent by email to: 
 
Roberto Pili and Ashley Orillion at rpili@iupui.edu and aorillio@iu.edu. 
 
Attn: Ashley Orillion 
Laboratory of Dr. Roberto Pili 
Division of Hematology and Oncology, 
Department of Medicine, 
Indiana University School of Medicine 
Joseph E. Walther hall 
Pharmacokinetic studies (Phase I only) 
980 W. Walnut Street, R3 C550 
Indianapolis, IN 46202 
 
7.2 
Pharmacokinetic studies (Phase I only) 
 
Pharmacokinetic analysis will be performed by Michelle A. Rudek, PharmD, PhD. at the 
Analytical Pharmacology Core Laboratory at the SKCCC at Johns Hopkins. Patients in the phase 
I component of the study will have plasma samples collected to assess entinostat 
pharmacokinetics. Entinostat concentrations will be assessed pretreatment on Day -14 and at 2 
and 6 hrs., Day -7, pretreatment on Day 1 and at 2 and 6 hrs. and Day 15 during the first cycle 
and at the time of progression. These pharmacokinetic parameters will be used to explore the 
relationship between entinostat exposure with pharmacodynamic (PD) endpoints such as toxicity 
and histone acetylation in peripheral blood mononuclear cells or PBMCs and changes in T cell 
subset populations. 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 32 of 78 
 
 
Blood sample preparation 
Blood will be collected for entinostat pharmacokinetics in sodium heparin vacutainer tubes. Each 
sample will require 5 mL of patient blood. Samples will be placed on ice immediately and 
centrifuged within 30 minutes of collection at 1,000 xg for 10 minutes in a refrigerated 
centrifuge. Plasma will be stored at –20°C until analysis. Entinostat concentrations will be 
determined using a validated LC/MS/MS method that will be developed by the Analytical 
Pharmacology Core Laboratory (Zhao M, Rudek MA, Mnasakanyan A, Hartke C, Pili R, Baker 
SD.A liquid chromatography/tandem mass spectrometry assay to quantitate entinostat in human 
plasma.J Pharm Biomed Anal. 2007 Jan 17;43(2):784-7.). 
 
Sample Shipment  
All specimens should be correctly labeled with patient initials, study-specific subject ID number, 
protocol number, and date and time of collection. NOTE: In the event that baseline specimens 
are available but the patient is not successfully registered to the protocol, do not submit the 
patient's specimens. Overnight shipments should occur on Monday through Wednesday except 
when the following day is a holiday. A fax or call should be placed to the Analytical 
Pharmacology Core Laboratory prior to shipment providing the shipment tracking information. 
 
Address shipments and any question regarding specimen processing to: 
 
Analytical Pharmacology Core Laboratory  
Attn: Entinostat/IL-2 Study Samples 
Bunting-Blaustein Cancer Research Building 1 
1650 Orleans Street 
Room 184 
Baltimore, MD 21231-1000 
Phone: 410-955-1129 
Phone with voicemail: 410-502-7192 
Fax: 410-502-0895 
 
7.3 
Non invasive imaging studies 
 
Non invasive imaging studies assessing changes in tumor metabolism will be performed both at 
RPCI and JHU. This study will evaluate changes in tumor metabolism in all patients undergoing 
PET/CT utilizing 18FDG. Patients will have PET/CT studies before starting entinostat 
administration and during week 5. 
 
7.4 
Optional tumor Samples 
 
All Patients with accessible tumors will be asked to undergo an optional tumor biopsy within 21 
days before starting treatment and on Day 30 of Cycle 1 +/- 2 days. This tumor biopsy is not 
required for patients to take part in this protocol. 
 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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7.4.1. Formalin-Fixed Paraffin-Embedded Tissue 
Sample Collection Time Points 
 
Samples may be obtained from any time prior to drug dosing. 
 
 
Sample Collection Procedure 
Per institutional procedures, obtain a whole formalin-fixed paraffin block for patient. 
Paraffin blocks should contain a tumor specimen measuring at least 0.5 cm x 0.5 cm x 
100 microns. Paraffin blocks may be from original diagnosis or optional biopsy any 
time prior to first drug treatment. 
 
Packaging of Paraffin Block 
 
• Place the paraffin block in a specimen container. 
 
NOTE: If the entire paraffin block cannot be submitted, please submit 10 
unstained slides to:  
Attn: Ashley Orillion 
Laboratory of Dr. Roberto Pili 
Division of Hematology and Oncology, 
Department of Medicine, 
Indiana University School of Medicine 
Joseph E. Walther hall 
980 W. Walnut Street, R3 C550 
Indianapolis, IN 46202 
 
 
Shipment of Paraffin-embedded Tissue 
 
Notification of sample shipment should be sent by email to 
Roberto Pili and Ashley Orillion at rpili@iupui.edu and aorillio@iu.edu. 
Analysis 
These samples will be analyzed for microvessel density (anti-CD34 staining), HIF-1  staining, 
acetylated Histone and CD4+/FOXp3+ T cells. All IHC studies will be done with established 
protocols in Dr. Roberto Pili’s lab. 
 
 
7.4.2 Optional fresh tumor samples 
Sample Collection  
• Optional tumor biopsy samples will be collected from patients with 
accessible tumor per institutional procedure. 
Sample 
Screening Visit 
Original diagnostic material 
X 
Any subsequent biopsy material prior to dosing 
X 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 34 of 78 
 
• The specific biopsy samples and time points for collection of fresh tumor 
samples are given below: 
 
Time Points 
Number of Tumor Biopsy Samples  
per Time Point 
Pretreatment 
1 
On treatment* 
1 
 
* The on-treatment samples will be collected 2 hours following the most recent dose of 
entinostat on Day 30 Cycle 1 +/- 2 days. To ensure that these samples are obtained at the 
correct time, patients are urged to take their entinostat dose in the clinic on the day of on-
treatment sample collection. The time of the entinostat dose and sample collection must be 
recorded. 
 
Attn: Ashley Orillion 
Laboratory of Dr. Roberto Pili 
Division of Hematology and Oncology, 
Department of Medicine, 
Indiana University School of Medicine 
Joseph E. Walther hall 
980 W. Walnut Street, R3 C550 
Indianapolis, IN 46202 
 
Notification of sample shipment should be sent by email to 
Roberto Pili and Ashley Orillion at rpili@iupui.edu and aorillio@iu.edu. 
These samples will be also analyzed for microvessel density (anti-CD34 staining), HIF-1 
 staining, acetylated histone, CD4+, CD4+/FOXp3+, and CD8+ T cells. All IHC studies will be 
done with established protocols in Dr. Roberto Pili’s lab. 
 
 

Entinostat and Aldesleukin 
                           PI: Saby George, M.D 
 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
 page 35 of 78 
 
8. STUDY CALENDAR 
 
Pre-study evaluations are to be performed within 4 weeks prior to start of therapy.  
Treatments and 
Evaluations 
Pre-
Study 
Cycle 1 and subsequent cycles 
Every 4 
weeks  
between  
cycles8 
 
End of 
Treat 
ment9 
Follow-
Up9a 
Week -2 
Week 1 
Week 3 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
D 
-14 
-13 -12 
-11 -10 -9 
-7 
1 
2 
3 
4 
5 
6 
7 
15 
16 
17 
18 
19 
20 
21 
Entinostat dosing 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
 
 
 
IL-2 dosing 
 
 
 
 
 
 
 
 
√ 
√ 
√ 
√ 
√ 
 
 
√10 
√ 
√ 
√ 
√ 
 
 
 
 
 
Informed Consent 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
History and Physical 
√ 
 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√8a 
√ 
 
Vital Signs and Weight 
√ 
√11 
 
 
 
 
 
√ 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√8a 
√ 
 
ECOG Performance Status 
√ 
√11 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√8a 
√ 
 
CBC w/diff., plts 
√ 
√11 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
√ 
 
Urine analysis, (CMP)1 
√ 
 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
√ 
 
PT/PTT/INR 
√ 
 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
√ 
 
Serum Pregnancy test2 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Head MRI or CT 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
EKG,MUGA, cardiac  
stress test3 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
PFT 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Tumor Measurements4 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
√4 
√ 
 
Pharmacodynamic  
Samples5 & 5a 
 
√11 
 
 
 
 
 
√ 
√ 
√ 
 
 
 
 
 
√ 
 
 
 
 
 
 
 
√ 
 
Serum for microRNA12 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
 
 
 
 
Tumor Needle Biospy6 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
PET/CT7 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Concomitant Meds 
√ 
√11 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
√ 
√ 
 
Survival follow-up 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
√ 
 
 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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1. Comprehensive metabolic panel (CMP) to include measurement of sodium potassium, chloride, bicarbonate/CO2, BUN, creatinine, glucose, total bilirubin, corrected calcium (CC= Serum 
calcium+0.8(4-Serum albumin), total proteins, albumin, AST, ALT, alkaline phosphatase, LDH, magnesium, and phosphate. 
2. Pregnancy test required within 14 days of starting study drug for women of child bearing potential only. 
3. Patients who are over 40 or have had previous myocardial infarction greater than 6 months prior to entry will be required to have a negative or low probability cardiac stress test for  
 cardiac ischemia, as well as EKG and MUGA. Subsequent cardiac tests may be performed at the discretion of the Principal Investigator as clinically indicated.  
4. Include CT scan, bone scan, MRI or ultrasound (additional tests may be performed as clinically indicated) Perform on week 11 of each cycle (+/- 7days) and every 8-12 weeks (+/-  
 2 weeks) on entinostat. Bone scan is required at baseline for patients with bone pain or evidence of bone lesions on CT scan. 
5. Obtain pre-dose on day -14; day -7; pre-dose day 1 and at 8 hrs and 24 hrs post dose of entinostat; and pre-dose day 15 during the first cycle and at the time of progression. 
 5a. Obtain PD sample on cycle 2 pre-dose day 1 and day 15 pre-dose; Repeat for cycle 3 pre-dose day 1 and day 15 pre-dose. 
6. Optional Core biopsy and fine needle aspirate obtained within 21 days prior to start of cycle 1 and on day 30 of cycle 1 +/- 2 days. 
7. To be performed prior to cycle 1 and on week 5 to assess changes in tumor blood metabolism. 
8. Maintenance Period, Local labs results acceptable. 
8a. May be performed every 8 – 12 weeks between cycles. 
9. Performed 30 days (+/- 4 days) following last dose of study drug. 
 9a. every three months afterward assessment may be obtained via telephone. 
10. Beginning of course 2 (day 15) may be delayed up to 7 days. 
11. Must be performed within 3 days prior to day -14. 
12. Obtain Serum for microRNA for cycles 2 an 3 on day 1pre-dose and at time of progression 
 
 

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9. MEASUREMENT OF EFFECT 
 
Patients will undergo CT scans at week 11 (+/- 7 days) of each cycle during aldesleukin 
administration and then every 8-12 weeks (+/- 2 weeks). 
 
Response Evaluation Criteria in Solid Tumors (RECIST V.1.0) Quick Reference 
Only patients with measurable disease at baseline should be included in protocols where 
objective tumor response is the primary endpoint. 
 
Measurable disease - the presence of at least one measurable lesion. If the measurable 
disease is restricted to a solitary lesion, its neoplastic nature should be confirmed by 
cytology/histology. 
 
Measurable lesions - lesions that can be accurately measured in at least one dimension 
with longest diameter 20 mm using conventional techniques or 10 mm with spiral CT 
scan. 
 
Non-measurable lesions - all other lesions, including small lesions (longest diameter <20 
mm with conventional techniques or <10 mm with spiral CT scan), i.e., bone lesions, 
leptomeningeal disease, ascites, pleural/pericardial effusion, inflammatory breast disease, 
lymphangitis cutis/pulmonis, cystic lesions, and also abdominal masses that are not 
confirmed and followed by imaging techniques; and. 
 
• All measurements should be taken and recorded in metric notation, using a ruler or calipers. 
All baseline evaluations should be performed as closely as possible to the beginning of 
treatment and never more than 4 weeks before the beginning of the treatment.  
• The same method of assessment and the same technique should be used to characterize each 
identified and reported lesion at baseline and during follow-up.  
• Clinical lesions will only be considered measurable when they are superficial (e.g., skin 
nodules and palpable lymph nodes). For the case of skin lesions, documentation by color 
photography, including a ruler to estimate the size of the lesion, is recommended. 
 
9.1 
Methods of Measurement 
 
• CT and MRI are the best currently available and reproducible methods to measure target lesions 
selected for response assessment. Conventional CT and MRI should be performed with cuts of 10 
mm or less in slice thickness contiguously. Spiral CT should be performed using a 5 mm 
contiguous reconstruction algorithm. This applies to tumors of the chest, abdomen and pelvis. 
Head and neck tumors and those of extremities usually require specific protocols. 
 
• Lesions on chest X-ray are acceptable as measurable lesions when they are clearly defined 
and surrounded by aerated lung. However, CT is preferable. 
 
• When the primary endpoint of the study is objective response evaluation, ultrasound (US) 
should not be used to measure tumor lesions. It is, however, a possible alternative to clinical 

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measurements of superficial palpable lymph nodes, subcutaneous lesions and thyroid 
nodules. US might also be useful to confirm the complete disappearance of superficial 
lesions usually assessed by clinical examination. 
 
• The utilization of endoscopy and laparoscopy for objective tumor evaluation has not yet been 
fully and widely validated. Their uses in this specific context require sophisticated equipment 
and a high level of expertise that may only be available in some centers. Therefore, the 
utilization of such techniques for objective tumor response should be restricted to validation 
purposes in specialized centers. However, such techniques can be useful in confirming 
complete pathological response when biopsies are obtained. 
 
• Tumor markers alone cannot be used to assess response. If markers are initially above the 
upper normal limit, they must normalize for a patient to be considered in complete clinical 
response when all lesions have disappeared. 
 
• Cytology and histology can be used to differentiate between PR and CR in rare cases (e.g., 
after treatment to differentiate between residual benign lesions and residual malignant lesions 
in tumor types such as germ cell tumors). 
 
Baseline documentation of “Target” and “Non-Target” lesions 
 
• All measurable lesions up to a maximum of five lesions per organ and 10 lesions in total, 
representative of all involved organs should be identified as target lesions and recorded and 
measured at baseline. 
• Target lesions should be selected on the basis of their size (lesions with the longest diameter) 
and their suitability for accurate repeated measurements (either by imaging techniques or 
clinically). 
• A sum of the longest diameter (LD) for all target lesions will be calculated and reported as 
the baseline sum LD. The baseline sum LD will be used as reference by which to 
characterize the objective tumor. 
• All other lesions (or sites of disease) should be identified as non-target lesions and should 
also be recorded at baseline. Measurements of these lesions are not required, but the presence 
or absence of each should be noted throughout follow-up. 
 
9.2 
Response Criteria 
 
9.2.1 Evaluation of target lesions 
* Complete Response (CR): 
Disappearance of all target lesions 
* Partial Response (PR): 
At least a 30% decrease in the sum of the LD of target lesions, 
taking as reference the baseline sum LD 

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* Progressive Disease (PD): 
At least a 20% increase in the sum of the LD of target lesions, 
taking as reference the smallest sum LD recorded since the 
treatment started or the appearance of one or more new lesions 
* Stable Disease (SD): 
Neither sufficient shrinkage to qualify for PR nor sufficient 
increase to qualify for PD, taking as reference the smallest sum 
LD since the treatment started 
 
9.2.2 Evaluation of non-target lesions 
* Complete Response (CR): 
Disappearance of all non-target lesions and normalization of 
tumor marker level 
  
*Incomplete Response/ 
Stable Disease (SD): 
Persistence of one or more non-target lesion(s) or/and 
maintenance of tumor marker level above the normal limits 
* Progressive Disease (PD): 
Appearance of one or more new lesions and/or unequivocal 
progression of existing non-target lesions (1)  
(1) Although a clear progression of “non-target” lesions only is exceptional, in such 
circumstances, the opinion of the treating physician should prevail and the progression 
status should be confirmed later on by the review panel (or study chair). 
 
9.2.3 Evaluation of best overall response 
 
The best overall response is the best response recorded from the start of the treatment until 
disease progression/recurrence (taking as reference for PD the smallest measurements recorded 
since the treatment started). In general, the patient's best response assignment will depend on the 
achievement of both measurement and confirmation criteria. 
 
Target lesions 
Non-Target lesions 
New Lesions 
Overall response 
CR 
CR 
No 
CR 
CR 
Incomplete 
response/SD 
No 
PR 
PR 
Non-PD 
No 
PR 
SD 
Non-PD 
No 
SD 

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PD 
Any 
Yes or No 
PD 
Any 
PD 
Yes or No 
PD 
Any 
Any 
Yes 
PD 
 
• Patients with a global deterioration of health status requiring discontinuation of treatment 
without objective evidence of disease progression at that time should be classified as having 
“symptomatic deterioration”. Every effort should be made to document the objective 
progression even after discontinuation of treatment. 
 
• In some circumstances it may be difficult to distinguish residual disease from normal tissue. 
When the evaluation of complete response depends on this determination, it is recommended 
that the residual lesion be investigated (fine needle aspirate/biopsy) to confirm the complete 
response status. 
 
9.2.4 Confirmation 
 
• The main goal of confirmation of objective response is to avoid overestimating the response 
rate observed. In cases where confirmation of response is not feasible, it should be made 
clear when reporting the outcome of such studies that the responses are not confirmed. 
 
• To be assigned a status of PR or CR, changes in tumor measurements must be confirmed by 
repeat assessments that should be performed no less than 4 weeks after the criteria for 
response are first met. Longer intervals as determined by the study protocol may also be 
appropriate.  
 
• In the case of SD, follow-up measurements must have met the SD criteria at least once after 
study entry at a minimum interval (in general, not less than 6-8 weeks) that is defined in the 
study protocol  
 
9.2.5 Duration of overall response 
 
• The duration of overall response is measured from the time measurement criteria are met for 
CR or PR (whichever status is recorded first) until the first date that recurrence or PD is 
objectively documented, taking as reference for PD the smallest measurements recorded 
since the treatment started. 
 
9.2.6 Duration of stable disease 
 
• SD is measured from the start of the treatment until the criteria for disease progression are 
met, taking as reference the smallest measurements recorded since the treatment started. 
 
• The clinical relevance of the duration of SD varies for different tumor types and grades. 

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Therefore, it is highly recommended that the protocol specify the minimal time interval 
required between two measurements for determination of SD. This time interval should take 
into account the expected clinical benefit that such a status may bring to the population under 
study.  
 
9.2.7 Reporting of results 
 
• All patients included in the study must be assessed for response to treatment, even if there are 
major protocol treatment deviations or if they are ineligible. Each patient will be assigned 
one of the following categories: 1) complete response, 2) partial response, 3) stable disease, 
4) progressive disease, 5) early death from malignant disease, 6) early death from toxicity, 7) 
early death because of other cause, or 8) unknown (not assessable, insufficient data). 
 
• All of the patients who met the eligibility criteria should be included in the main analysis of 
the response rate. Patients in response categories 4-8 should be considered as failing to 
respond to treatment (disease progression). Thus, an incorrect treatment schedule or drug 
administration does not result in exclusion from the analysis of the response rate. Precise 
definitions for categories 4-8 will be protocol specific. 
 
• All conclusions should be based on all eligible patients. 
 
• Sub-analyses may then be performed on the basis of a subset of patients, excluding those for 
whom major protocol deviations have been identified (e.g., early death due to other reasons, 
early discontinuation of treatment, major protocol violations, etc.). However, these sub-
analyses may not serve as the basis for drawing conclusions concerning treatment efficacy, 
and the reasons for excluding patients from the analysis should be clearly reported. 
 
• The 95% confidence intervals should be provided 
 
10 
REGULATORY AND REPORTING REQUIREMENTS 
 
10.1 
Protocol Amendments 
 
Any changes to the protocol will be made in the form of an amendment and must be approved by 
CTEP and the IRB before implementation. 
 
10.2 
Informed Consent 
 
An investigator will explain to each subject (or legally authorized representative) the nature of 
the study, its purpose, procedures involved, expected duration, potential risks and benefits. Each 
subject will be informed that participation in the study is voluntary and that he may withdraw 
from the study at any time, and that withdrawal of consent will not affect her subsequent medical 
treatment. This informed consent will be given by means of a standard written statement and will 
be submitted for IRB approval prior to use. No patient will enter the study before her informed 

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consent has been obtained. 
 
10.3 
Adverse Events 
 
Expedited adverse event (AE) reporting for this study is via CTEP-AERS (CTEP Adverse Event 
Reporting System), accessed via the secure CTEP website:  
(https://eapps-ctep.nci.nih.gov/ctepaers/pages/task?rand). The reporting procedures are presented 
in the “CTEP, NCI Guidelines: Adverse Event Reporting Requirements” and can be downloaded 
from: 
(http://ctep.cancer.gov/protocolDevelopment/elcetronicapplications/docs/aeguidelines.pdf). 
 
The descriptions and grading scales found in the CTEP Active Version of the NCI Common 
Terminology Criteria for Adverse Events (CTCAE) will be utilized for adverse event reporting. 
The CTEP Active Version of the CTCAE is identified and located on the CTEP website at: 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm. 
All 
appropriate 
treatment areas should have access to a copy of the CTEP Active Version of CTCAE. A list of 
adverse events that have occurred or might occur (Comprehensive Adverse Events and Potential 
Risks list) can be found in Section 6 (Pharmaceutical Information). A copy of the current Agent-
Specific Adverse Event List (ASAEL) is sent with LOI approval letters. The ASAEL is updated 
periodically; copies of the updated list are available on request via e-mail from aemd@tech-
res.com. 
 
Information about all adverse events, whether volunteered by the subject, discovered by 
investigator questioning, or detected through physical examination, laboratory test or other 
means, will be collected, recorded, and followed as appropriate. 
 
All adverse events experienced by subjects will be collected from the time of consent, 
throughout the study, and until the final study visit. Subjects may be contacted for additional 
assessments, as necessary. 
 
Definitions  
Adverse Event (AE): Any undesirable sign, symptom or medical condition occurring after 
starting study drug (or therapy) even if the event is not considered to be related to the study. 
 
Medical conditions/diseases present before starting study treatment are only considered adverse 
events if they worsen after starting study treatment (any procedures specified in the protocol). 
Adverse events occurring before starting study treatment but after signing the informed consent 
form will be recorded. Abnormal laboratory values or test results constitute adverse events only 
if they induce clinical signs or symptoms, require therapy or as required by SPEER. 
 
Serious adverse event or reaction: Any untoward medical occurrence that at any dose: results in 
death, is life-threatening, requires inpatient hospitalization or prolongation of existing 
hospitalization, results in persistent or significant disability / incapacity, or is a congenital 
anomaly / birth defect. 
 
The definition of serious adverse event (experience) also includes important medical event. 

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Medical and scientific judgment should be exercised in deciding whether expedited reporting is 
appropriate in other situations, such as important medical events that may not be immediately 
life-threatening or result in death or hospitalization but may jeopardize the patient or may require 
intervention to prevent one of the other outcomes listed in the definition above. These should 
also usually be considered serious. Examples of such events are intensive treatment in an 
emergency room or at home for allergic bronchospasm; blood dyscrasias or convulsions that do 
not result in hospitalization; or development of drug dependency or drug abuse. 
 
Unexpected adverse event: An adverse event, which varies in nature, intensity or frequency from 
information on the investigational drug/agent provided in the Investigator’s Brochure, 
package insert or safety reports. Any adverse event that is not included in the informed 
consent is considered “unexpected”. 
 
Expected (known) adverse event: An adverse event, which has been reported in the 
Investigator’s Brochure. An adverse event is considered “expected”, only if it is 
included in the informed consent document as a risk. 
 
Relationship 
 
The relationship of all adverse events and serious adverse events to study medication will be 
assigned as follows: 
Definitely: An adverse event which has a timely relationship to the administration of the 
investigational drug/agent, follows a known pattern of response, for which no alternative cause is 
present 
 
Probably: An adverse event, which has a timely relationship to the administration of the 
investigational drug/agent, follows a known pattern of response, but for which a potential 
alternative cause may be present 
 
Possibly: An adverse event, which has a timely relationship to the administration of the 
investigational drug/agent, follows no known pattern of response, but a potential alternative 
cause does not exist. 
 
Unrelated: An adverse event, for which there is evidence that it is definitely related to a cause 
other than the investigational drug/agent. In general, there is no timely relationship to the 
administration of the investigational drug/agent, or if there is a timely relationship, the event 
does not follow a known pattern of response, and there is an alternative cause. 
 
10.3.1 Reporting Procedures 
 
General 
All adverse events will be captured on the appropriate study-specific case report forms (CRFs). 
All serious adverse events, regardless of causality to study drug, will be reported promptly to the 
Principal Investigator and/or the Study Coordinator. Participating network sites refer to appendix 
D for reporting instructions. 
 

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Institutional Review Board (IRB) 
All adverse events/serious adverse events will be reported to the IRB per current institutional 
standards. If an adverse event requires modification of the study protocol and informed consent, 
these modifications will be provided to the IRB with the report of the adverse event. 
 
External Agencies 
 
Expedited Adverse Event Reporting – NCI/CTEP 
Expedited reports are submitted to CTEP via the secure CTEP-AERS application accessed via 
the CTEP web site 
(https://eapps-ctep.nci.nih.gov/ctepaers/pages/task?rand). Those AEs that do not require 
expedited reporting must be reported in routine (CDUS) study data submissions. AEs reported 
through CTEP-AERS must also be reported in routine study data submissions. 
 
In the rare event when Internet connectivity is disrupted a 24-hour notification is to be made to 
NCI by telephone at: 301-897-7497 or 301-897-7402 for CIP studies or fax to 301-230-0159. An 
electronic report must be submitted immediately upon re-establishment of the internet 
connection. 
 
Expedited Reporting Guidelines: Phase 1 Trials Utilizing an Agent under a CTEP IND: CTEP-
AERS Reporting Requirements for Adverse Events That Occur Within 30 Days1 of the Last 
Dose of the Investigational Agent. 
 
 
 
Grade 1 
 
Grade 2 
 
Grade 2 
 
Grade 3 
 
Grade 3 
Grades 
4 & 52 
Unexpected  
and Expected 
 
Unexpected 
 
 
Expected 
Unexpected 
Expected 
Unexpected  
and 
Expected 
with  
Hospitali-
zation 
without 
Hospitali- 
zation 
with  
Hospitali- 
zation 
without 
Hospitali- 
zation 
Unrelated 
Unlikely 
Not 
Required 
Not 
Required 
Not 
Required 
10 
Calendar 
Days 
Not 
Required 
10 
Calendar 
Days 
Not 
Required 
24-Hour; 
5 Calendar 
Days 
Possible 
Probable 
Definite 
Not 
Required 
10 Calendar 
Days 
Not 
Required 
24-Hour; 
5 Calendar 
Days 
24-Hour; 
5 Calendar 
Days 
10 
Calendar 
Days 
Not 
Required 
24-Hour; 
5 Calendar 
Days 
1 Adverse events with attribution of possible, probable, or definite that occur greater than 30 days after the last dose of 
treatment with an agent under a CTEP IND require reporting as follows: 
CTEP-AERS 24-hour notification followed by complete report within 5 calendar days for: 
• 
Grade 3 unexpected events with hospitalization or prolongation of hospitalization 
• 
Grade 4 unexpected events  

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• 
Grade 5 expected events and unexpected events 
 
2 Although a CTEP-AERS 24-hour notification is not required for death clearly related to progressive disease, a full report is 
required as outlined in the table. 
December 15, 2004 
Note: All deaths on study require both routine and expedited reporting regardless of causality. 
Attribution to treatment or other cause must be provided. 
 
• Expedited AE reporting timelines defined: 
➢ “24 hours; 5 calendar days” – The investigator must initially report the AE via CTEP-
AERS within 24 hours of learning of the event followed by a complete CTEP-AERS 
report within 5 calendar days of the initial 24-hour report. 
➢ “10 calendar days” - A complete CTEP-AERS report on the AE must be submitted 
within 10 calendar days of the investigator learning of the event.  
 
• Any medical event equivalent to CTCAE grade 3, 4, or 5 that precipitates hospitalization 
(or prolongation of existing hospitalization) must be reported regardless of 
attribution and designation as expected or unexpected with the exception of any 
events identified as protocol-specific expedited adverse event reporting exclusions. 
 
• Any event that results in persistent or significant disabilities/incapacities, congenital 
anomalies, or birth defects must be reported via CTEP-AERS if the event occurs 
following treatment with an agent under a CTEP IND. 
 
• Use the NCI protocol number and the protocol-specific patient ID provided during trial 
registration on all reports. 
 
 
 

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Phase 2 Trials Utilizing an Agent under a CTEP IND: CTEP-AERS Reporting Requirements for 
Adverse Events That Occur Within 30 Days1 of the Last Dose of the Investigational Agent 
 
 
 
Grade 1 
 
Grade 2 
 
Grade 2 
 
Grade 3 
 
Grade 3 
Grades 
4 & 52 
Grades 
4 & 52 
Unexpected 
and 
Expected 
 
Unexpected 
 
Expected 
Unexpected 
Expected 
Unexpected  Expected 
with 
Hospitali-
zation 
Without 
Hospitali
-zation 
with 
Hospitalization 
without 
Hospitalization 
Unrelated 
Unlikely 
Not 
Required 
Not 
Required 
Not 
Required 
10 
Calendar 
Days 
Not 
Required 
10 Calendar 
Days 
Not 
Required 
10 
Calendar 
Days 
10 
Calendar 
Days 
Possible 
Probable 
Definite 
Not 
Required 
10 Calendar 
Days 
Not 
Required 
10 
Calendar 
Days 
10 
Calendar 
Days 
10 Calendar 
Days 
Not 
Required 
24-Hour; 
5 Calendar 
Days 
10 
Calendar 
Days 
1 Adverse events with attribution of possible, probable, or definite that occur greater than 30 days after the last dose of 
treatment with an agent under a CTEP IND require reporting as follows: 
CTEP-AERS 24-hour notification followed by complete report within 5 calendar days for: 
• 
Grade 4 and Grade 5 unexpected events  
 
 CTEP-AERS10 calendar day report: 
• 
Grade 3 unexpected events with hospitalization or prolongation of hospitalization 
• 
Grade 5 expected events 
 
2 Although a CTEP-AERS 24-hour notification is not required for death clearly related to progressive disease, a full report is required as 
outlined in the table. 
December 15, 2004 
Note: All deaths on study must be reported using expedited reporting regardless of causality. 
Attribution to treatment or other cause should be provided. 
 
• 
Expedited AE reporting timelines defined: 
➢ “24 hours; 5 calendar days” – The investigator must initially report the AE via CTEP-
AERS within 24 hours of learning of the event followed by a complete CTEP-AERS 
report within 5 calendar days of the initial 24-hour report. 
➢ “10 calendar days” - A complete CTEP-AERS report on the AE must be submitted 
within 10 calendar days of the investigator learning of the event. 
 
• Any medical event equivalent to CTCAE grade 3, 4, or 5 that precipitates hospitalization 
(or prolongation of existing hospitalization) must be reported regardless of 

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attribution and designation as expected or unexpected with the exception of any 
events identified as protocol-specific expedited adverse event reporting exclusions. 
 
• Any event that results in persistent or significant disabilities/incapacities, congenital 
anomalies, or birth defects must be reported via CTEP-AERS if the event occurs 
following treatment with an agent under a CTEP IND. 
 
• Use the NCI protocol number and the protocol-specific patient ID provided during trial 
registration on all reports. 
 
Expedited Adverse Event Reporting Exclusions – Not Applicable 
 
10.3.2 Unanticipated Problems 
 
Definition 
An Unanticipated Problem is any unexpected event that affects the rights, safety or welfare of 
subjects or others that result from the research. This may involve physical, social, psychological, 
legal or other risks. 
 
An event is an Unanticipated Problem when it fulfills the definition above; it must be related or 
possibly related to the research and it must be Serious (see section 8.2). Unanticipated is defined 
as the nature, severity, or frequency of the event is not consistent with the current Investigator’s 
Brochure, protocol or alternative risk information and is not due to the progression of a disease 
process.  
 
An Unanticipated Problem (UAP) is any incident, experience, or outcome that meets all of the 
following criteria: 
 
1. Unexpected (in terms of nature, severity, or frequency) given: 
(a) the research procedures that are described in the study- related documents, including study 
deviations, as well as issues related to compromise of patient privacy or confidentiality of 
data;  
(b) the characteristics of the subject population being studied 
 
2. 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) 
 
3. Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or recognized 
and deemed Serious. 
 
10.3.2.1 Reporting Unanticipated Problems 
The principal investigator or delegated research staff will submit an Unanticipated 
Problem Form to the CRS Compliance Office via Fax: 716-845-8815 or email: 

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CRSCompliance for all unanticipated problems and adverse events that are serious and 
deemed related or possibly related to the research drug, biologic or intervention. 
 
10.3.2.2 Secondary AML/MDS 
All secondary malignancies that occur following treatment with an agent under an 
NCI/IDE must be reported via CTEP-AERS. CTCAE v4.0 has three options available 
to describe treatment-related events: 
 
• Leukemia secondary to on oncology chemotherapy. 
• Myelodysplastic syndrome 
• Treatment related secondary malignancy 
 
If you are reporting in CTCAE v3.0, the event can be reported as: “Secondary 
malignancy-Other (specify)”. 
 
10.4 Data and Safety Monitoring 
 
Scheduled meetings will take place monthly and include the protocol principal investigator, 
study coordinators, research nurses, and, when appropriate, the collaborators, sub-investigators, 
and biostatistician involved with the conduct of the protocol. During these meetings the 
investigators will discuss matters related to: safety of protocol participants, validity and integrity 
of the data, enrollment rate relative to expectation, characteristics of participants, retention of 
participants, adherence to protocol (potential or real protocol violations), data completeness, and 
progress of data for secondary objectives such as surrogate markers. A review of all grade 3/4 
toxicities will take place in each of the meetings as well as any delays in surgical procedure or in 
wound healing that may be contributed to study drug. 
 
Quarterly reports will be sent to CTEP according to specified CTEP guidelines. 
 
10.5 CTEP Multicenter Guidelines 
 
10.5.1 Responsibilities of the Principal Investigator. 
 
• The Principal Investigator will be the single liaison with the CTEP Protocol and 
Information Office (PIO). The Principal Investigator is responsible for the coordination, 
development, submission, and approval of the protocol as well as its subsequent 
amendments. The protocol must not be rewritten or modified by anyone other than the 
Principal Investigator. There will be only one version of the protocol, and each 
participating institution will use that document. The Principal Investigator is responsible 
for assuring that all participating institutions are using the correct version of the protocol. 
 
• The Principal Investigator is responsible for the overall conduct of the study at all 
participating institutions and for monitoring its progress. All reporting requirements to 
CTEP are the responsibility of the Principal Investigator.  
 

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• The Principal Investigator is responsible for the timely review of Adverse Events (AE) to 
assure safety of the patients. 
 
• The Principal Investigator is responsible for the review of and timely submission of data 
for study analysis. 
 
10.5.2 Responsibilities of the Coordinating Center 
 
• Each participating institution will have an appropriate assurance on file with the Office 
for Human Research Protection (OHRP), NIH. The Coordinating Center is responsible 
for assuring that each participating institution has an OHRP assurance and must maintain 
copies of IRB approvals from each participating site.  
 
• Prior to the activation of the protocol at each participating institution, an OHRP form 310 
(documentation of IRB approval) must be submitted to the CTEP PIO. 
 
• The Coordinating Center assures that IRB approval has been obtained at the participating 
site prior to the first patient registration from that site. 
 
• The Coordinating Center is responsible for the preparation of all submitted data for 
review by the Protocol Chair. 
 
• The Coordinating Center maintains documentation of AE reports. The participating 
institution reports directly to CTEP with a copy to the Coordinating Center. The 
Coordinating Center will submit AE reports to the Protocol Chair for timely review. 
 
• Audits may be accomplished in one of two ways: (1) source documents and research 
records for selected patients are brought from participating sites to the Coordinating 
Center for audit, or (2) selected patient records may be audited on-site at participating 
sites. If the NCI chooses to have an audit at the Coordinating Center, then the 
Coordinating Center is responsible for having all source documents, research records, all 
IRB approval documents, NCI Drug Accountability Record forms, patient registration 
lists, response assessments scans, x-rays, etc. available for the audit. 
 
10.5.3 Patient Registration 
 
The coordinating Center serves as the central location for registering all patients enrolled in a 
multi-center trial. Refer to network appendix D for patient registering instructions. 
 
10.6 Cooperative Research and Development Agreement (CRADA)/Clinical Trials 
Agreement (CTA) 
 
The agent(s), supplied by CTEP, DCTD, NCI, used in this protocol is/are provided to the NCI 
under a Collaborative Agreement (CRADA, CTA) between the Pharmaceutical Company(ies) 
[hereinafter referred to as “Collaborator(s)”] and the NCI Division of Cancer Treatment and 
Diagnosis. Therefore, the following obligations/guidelines, in addition to the provisions in the 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 50 of 79 
 
 
AIntellectual Property Option to Collaborator@ contained within the terms of award, apply to the 
use of Agent(s) in this study: 
 
1. Agent(s) may not be used for any purpose outside the scope of this protocol, nor can 
Agent(s) be transferred or licensed to any party not participating in the clinical study. 
Collaborator(s) data for Agent(s) are confidential and proprietary to Collaborator(s) and shall 
be maintained as such by the investigators. The protocol documents for studies utilizing 
investigational agents contain confidential information and should not be shared or 
distributed without the permission of the NCI. If a copy of this protocol is requested by a 
patient participating on the study or patient’s family member, the individual should sign a 
confidentiality agreement. A suitable model agreement can be downloaded from 
http://ctep.cancer.gov. 
 
2. For a clinical protocol where there is an investigational Agent used in combination with 
(an)other investigational Agent(s), each the subject of different collaborative agreements, the 
access to and use of data by each Collaborator shall be as follows (data pertaining to such 
combination use shall hereinafter be referred to as "Multi-Party Data”.): 
 
a. NCI must provide all Collaborators with prior written notice regarding the existence 
and nature of any agreements governing their collaboration with NIH, the design of 
the proposed combination protocol, and the existence of any obligations that would 
tend to restrict NCI's participation in the proposed combination protocol. 
b. Each Collaborator shall agree to permit use of the Multi-Party Data from the clinical 
trial by any other Collaborator solely to the extent necessary to allow said other 
Collaborator to develop, obtain regulatory approval, or commercialize its own 
investigational agent. 
c. Any Collaborator having the right to use the Multi-Party Data from these trials must 
agree in writing prior to the commencement of the trials that it will use the Multi-
Party Data solely for development, regulatory approval, and commercialization of its 
own investigational agent. 
 
3. Clinical Trial Data and Results and Raw Data developed under a Collaborative Agreement 
will be made available exclusively to Collaborator(s), the NCI, and the FDA, as appropriate 
and unless additional disclosure is required by law or court order. Additionally, all Clinical 
Data and Results and Raw Data will be collected, used and disclosed consistent with all 
applicable federal statutes and regulations for the protection of human subjects, including, if 
applicable, the Standards for Privacy of Individually Identifiable Health Information set forth 
in 45 C.F.R. Part 164. 
 
4. When a Collaborator wishes to initiate a data request, the request should first be sent to the 
NCI, who will then notify the appropriate investigators (Group Chair for Cooperative Group 
studies, or PI for other studies) of Collaborator's wish to contact them. 
 
5. Any data provided to Collaborator(s) for phase 3 studies must be in accordance with the 
guidelines and policies of the responsible Data Monitoring Committee (DMC), if there is a 
DMC for this clinical trial. 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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6. Any manuscripts reporting the results of this clinical trial must be provided to CTEP for 
immediate delivery to Collaborator(s) for advisory review and comment prior to submission 
for publication. Collaborator(s) will have 30 days from the date of receipt for review. 
Collaborator shall have the right to request that publication be delayed for up to an additional 
30 days in order to ensure that Collaborator’s confidential and proprietary data, in addition to 
Collaborator(s)’s intellectual property rights, are protected. Copies of abstracts must be 
provided to CTEP for forwarding to Collaborator(s) for courtesy review as soon as possible 
and preferably at least three (3) days prior to submission, but in any case, prior to 
presentation at the meeting or publication in the proceedings. Press releases and other media 
presentations must also be forwarded to CTEP prior to release. Copies of any manuscript, 
abstract, and/or press release/ media presentation should be sent to: 
 
Regulatory Affairs Branch, CTEP, DCTD, NCI 
Executive Plaza North, Suite 7111 
Bethesda, Maryland 20892 
FAX 301-402-1584 
Email: anshers@mail.nih.gov 
 
The Regulatory Affairs Branch will then distribute them to Collaborator(s). No publication, 
manuscript or other form of public disclosure shall contain any of Collaborator’s 
confidential/proprietary information. 
 
11. STATISTICAL CONSIDERATIONS 
 
11.1 Study Design- 
 
This is a phase I/II study of high dose interleukin 2 in combination with the histone deacetylase 
inhibitor entinostat in patients with metastatic renal cell carcinoma. The Phase I portion of the 
study will be used to evaluate the safety and tolerability of high dose interleukin 2 in 
combination with entinostat. High dose interleukin 2 (IL-2) is the standard of care for this patient 
population. It is hoped that a minimum effective dose of entinostat would enhance immune 
response with little additional toxicity. Because response rate may not fully reflect the additional 
immune response attributable to entinostat and because IL-2 is the standard of care, a single-
stage design will be used in the Phase II portion of the study to estimate the efficacy of the drug 
combination. Secondary endpoints will be used to understand the effect of entinostat and IL-2 on 
progression free survival (PFS) and parameters measuring immune response. A sequential 
probability ratio test (SPRT) will be used to provide a statistical threshold for stopping the study 
in case there is evidence of a larger proportion of extended grade 4 toxicities than allowed. 
 
11.2 Phase I 
 
Primary Objective: 
High-dose interleukin (IL-2) is the current standard therapy for metastatic renal cell carcinoma 
and its toxicity in a phase III study has been reported5. The primary objective of this portion of 
the study is to determine a recommended dose of the biologic agent, entinostat, when combined 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 52 of 79 
 
 
with IL-2. The IL-2 dose level is 600,000 IU/kg, every 8 hours, IV. The current recommended 
dose of entinostat when administered as a single agent every second week continuously is 10 
mg/m2. The entinostat study dose levels are 1, 3 and 5 mg, every 2 weeks, PO with 3 mg chosen 
as the starting dose for this study. A DLT will be defined as extended grade 4 toxicity (duration 
of one week or more) during the first 45 days of treatment with the recommended dose being the 
highest dose at which 0 or 1 DLTs are observed. A DLT rate of ≤ 33% will be permitted for 
further evaluation in the Phase II portion of the study. 
 
The first dose level will have a minimum of 3 patients treated (unless the first 2 patients 
experience dose-limiting toxicity(s) (DLT) before 3rd patient is enrolled. If a DLT occurs in 1 
patient treated at the starting dose level, another 3 patients will be treated at this dose level. If 
DLTs occur in 2 patients in the first 6 patients, patients will be treated at a lower dose level 
(entinostat 1 mg PO every 2 weeks, dose level 0) and DLT will be evaluated similarly. If DLT 
occurs in 1/6 patients, dose level 1 will be considered the recommended Phase II dose. 
 
If no DLTs occur at the starting dose level 1, 3 or 4 patients will be treated at the next dose level 
(level 2) with 5 mg PO every 2 weeks (wait 45 days for DLT assessment on first 2 patients 
before 3rd patient is enrolled). If no DLTs occur at dose level 2, this dose level will be 
recommended for the phase II portion of the study. If a DLT occurs in 1 patient at dose level 2, 
another 3 or 4 patients will be treated at this dose level. If DLT occurs in 2/6 patients at dose 
level 2, dose level 1 will be the recommended for the Phase II. If a DLT occurs in 1/6 patients, 
dose level 2 will be considered the recommended Phase II dose. 
 
Patients who experience prolonged grade 4 toxicity (more than one week) and recover to ≤ 
Grade 1 (or to pretreatment baseline level toxicity) may continue treatment at the next entinostat 
lower level. Once the entinostat has been dose reduced, patients must remain at the reduced dose 
for the remainder of the study. Patients may have their dose de-escalated twice before being 
removed from the trial. Patients will be allowed to remain on the therapy provided that they are 
tolerating the treatment and do not develop progressive disease. No dose de-escalation for IL-2 
will be performed. 
 
Secondary Objective: 
The secondary objective of this portion of the study will describe the toxicity and tolerability of 
the combination of IL-2 and entinostat. An exact binomial proportion with a 95% confidence 
interval will be given for prolonged grade 4 toxicity. The frequency and grade of toxicities will 
be tabulated for each dose level. Summary statistics (mean, standard deviation, frequency) will 
be used to record the number of doses of IL-2 administered during the first cycle of therapy and 
the toxicity after the scheduled 9th dose IL-2. 
 
11.3 Phase II 
 
Primary Objective: 
Once a recommended dose has been determined in the Phase I portion of the study, the cohort 
will be expanded so that 36 new patients will be treated at this dose level. The primary objective 
of this phase is to evaluate the safety and efficacy of the combination of entinostat and IL-2. The 
primary endpoint is overall response rate (complete plus partial). Enrollment is expected to be 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 53 of 79 
 
 
between 1 and 2 patients per month. The combination treatment will be considered unsuccessful 
if the response rate is 20% or less, and it will be considered active enough to pursue further if the 
response rate was 40% or greater. To test this hypothesis, the fixed sample size for a single-stage 
study with a type I error of 10% and a type II error of 10%, based on an exact binomial test, is 
36. If 11 or more of the patients have a response, the hypothesis that the response rate is ≤ 20% is 
rejected with a target error rate of 0.10. This design has type I and type II errors of 0.09 and 0.09, 
respectively. 
 
Secondary Objectives: 
Secondary endpoints will be used to determine the effect of entinostat on progression free 
survival (PFS), survival, time-to tumor progression and parameters measuring immune response. 
 
Failure rates will be used to compare the PFS, overall survival and time-to-tumor progression of 
metastatic RCC patients treated with high dose aldesleukin combined with entinostat to the 
historical failure rates of patients treated with high dose aldesleukin alone. The failure rate is 
estimated by dividing the number of events ( d ) by the sum of the exposure time in the study 
cohort: 

=
it
d /

. The 95% confidence interval is calculated as: 
d
Z



2
/
1−

.  
The historical PFS failure rate observed in the randomized trial of IL-2 reported by the Cytokine 
Working Group5 was 2.77 per person year (median PFS of 3 months). Patients eligible for this 
study are expected to be homogenous group with comparable prognostic characteristics. 
Enrollment is expected to be between 1 and 2 patients per month with an accrual period of 6 
years and 12 months of follow-up. With a total of 34 events in 36 patients, the study would yield 
79% power to detect a decrease in the PFS failure rate to 1.85 per person year of follow-up (an 
increase of 1.5 months in the median PFS compared to the historical median) at an alpha level of 
0.1 (one-sided). 
 
 
We do not expect that different doses of entinostat during the period off aldesleukin 
(maintenance) will affect the primary endpoint since objective responses have been rarely 
observed with single agent entinostat. We expect that the vast majority of patients will be able to 
tolerate the 10 mg maintenance dose if the phase II recommended dose in combination with 
aldesleukin is 5 mg. Should we observe discrepancies in the maintenance dose level of entinostat 
we will consider subgroup analysis to determine the secondary objective PFS in the different 
group of patients. 
 
 
The toxicity of IL-2 when combined with entinostat will be further characterized following the 
phase II portion of this study. The exact binomial proportion for prolonged grade 4 toxicities 
with the 95% confidence interval will be given. Additional toxicity frequencies, proportions, and 
95% CIs will be given by type and grade of toxicity. 
 
 
The association between responses (e.g. response, progression free and overall survival) and 
baseline laboratory parameters (e.g. CD4+, CD4+/Foxp3, CD8+, Ki 67, Tunel) will be 
summarized graphically (boxplots, scatterplots, Kaplan-Meier curves) and numerically (means, 
medians). For binary outcomes such as response, logistic regression will be used to assess the 
significance of associations individually (univariate) and while adjusting for other variables 
(multivariate). The Cox proportional hazards model will be used to model the effect of these 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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parameters on time-to-event outcomes. 
 
 
Logistic regression will be used to explore the relationship between entinostat exposure (plasma 
concentrations) with pharmacodynamic (PD) endpoints (e.g., toxicity and histone acetylation in 
peripheral blood mononuclear cells and change in T cell subset population). 
 
 
We will determine if initial levels of specific T lymphocytes (Tregs) or changes in the level of 
specific T lymphocytes from baseline might predict for response to this combination therapy. In 
this study Tregs are defined as CD4+CD25hi T cells. The hypotheses are that low baseline levels 
of Treg would be associated with an increased probability of response and that decreasing Tregs 
from baseline would be associated with an increased probability of response. Continuous 
baseline values of Treg (expressed as a frequency) and the change in Treg from baseline (4 
weeks post treatment – pretreatment) as well as binary versions of these variables will be 
explored for an association with response. A 30% decrease from baseline will be used as the 
binary cutpoint for Treg change. 
 
Two PET-derived indices of glucose metabolism: Standard Uptake Value (SUV) and FDG influx 
rate constant (Ki) will be used to determine whether early changes in tumor metabolisms by 
FDG PET/CT scan anticipate anatomic changes in patients receiving the combination of 
entinostat and IL-2. These two parameters will be measured at baseline and at week 5 using 
identical PET/CT acquisition protocols. Continuous change from baseline and binary (30% 
decrease from baseline) measures of these predictors will also be explored for an association 
with response. The hypothesis for these predictors is that treatment would decrease glucose 
metabolism, increasing response. 
 
Data analysis for Treg and PET correlative studies: 
This study will provide estimates of means and standard deviations for baseline Treg, change in 
Treg at week four and changes in the PET glucose metabolism indices at week five. 
Transformations of these predictors will be considered before analyses. For each endpoint, we 
will determine how well the responders and non-responders are distinguished. For binary 
predictors, the sensitivity and specificity with 95% confidence intervals will be reported. T tests 
will be used to compare the mean change between responders and non-responders. If there are 
sufficient numbers of responders, partial responders and non-responders an ANOVA will be 
used to compare changes in these three groups. If complete data are obtained for CD4+CD25hi T 
cells at multiple time points post treatment, repeated measures ANOVA will be performed to 
evaluate data for trends over time. 
 
11.4 Toxicity monitoring 
 
A sequential probability ratio test (SPRT) will be used to provide a monitoring boundary for 
stopping the study in case there is moderate evidence that the proportion of extended grade 4 
toxicities is 20% as opposed to the permitted rate of 5%. This monitoring boundary will be used 
to evaluate the toxicity data after every patient. The SPRT is based on testing two simple 
hypotheses on the basis of the likelihood ratio. The likelihood function for a binary toxicity 
outcome is binomial and the log likelihood ratio will be used to determine the relative evidence 
in favor of two hypothesized prolonged grade 4 toxicity rates, p1=0.2 and p2=0.05. The 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 55 of 79 
 
 
likelihood ratio is: 
 
Likelihood ratio (For p1 vs p2 | Data) = 
)
05
.0
|
Pr(
)
2.0
|
Pr(
=
=


data
data
=
x
N
x
x
N
x
−
−
−
−
)
1(
)
1(




  
where N=36 and x is the number of toxicities observed. We will stop the study if there is 
moderate evidence, LR≥8.0, that the proportion of prolonged grade 4 toxicities is 20% versus 
5%. The stopping boundary corresponding to this LR is given below. If the number of 
toxicities falls above the line any point along the study, the study will be stopped. Table 1 
gives the observed number of prolonged toxicities for which this boundary would be crossed 
after each patient accrued (N). The rate of erroneously stopping the study using this boundary 
is approximately 1/8 or 0.125. 
 
 
Table 1. Observed number of prolonged toxicities (Tox) where SPRT boundary would be 
crossed after each patient accrued (N). 
 
N 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 11 12 13 14 15 16 17 18 
Tox 2 
2 
2 
2 
2 
2 
3 
3 
3 
3 
3 
3 
3 
3 
3 
4 
4 
4 
 
N 
19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 
Tox 4 
4 
4 
4 
4 
4 
5 
5 
5 
5 
5 
5 
5 
5 
5 
6 
6 
6 
 
11.5 Patient Compliance 
 
All patients will receive study drug under direct observation by the site personnel. Dosing date 
5
10
15
20
25
30
35
Number
of
subjects
2
4
6
8
10
r
e
b
m
u
N
f
o
s
e
i
t
i
c
i
x
o
T

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Protocol Version Date  5/29/19 
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and time will be recorded by the site staff on the CRF. For all other doses, the patient will self-
administer his/her dose of oral medications per schedule and record the date and time in the 
patient diary. If a patient regurgitates any tablets, the patient should not repeat that dose, but 
should resume his/her scheduled subsequent doses. 
 
Pill counts will be utilized to monitor compliance as well as the patient diary. Patients will be 
supplied with a diary in which to record the date and time of each self-administered dose of 
study drugs. Patients will also be asked to record any partial doses. Study site staff will review 
the diary with the patient at each scheduled clinic visit and transcribe all pertinent information 
onto the CRF. The diaries will be retained at the study site as part of the source documentation 
and new diary pages issued to the patient following each visit. Patients will be asked to return all 
study medication bottles (empty, partial of full) to the study staff. 
 
11.6 Data quality assurance 
 
This study will be monitored regularly. Monitoring procedures include one or more visits 
designed to clarify all prerequisites before the study commences. Interim monitoring visits will 
take place on a regular basis according to a schedule fixed by mutual agreement. During these 
visits, the CRA will check for completion of the entries on the CRFs, their compliance with the 
study protocol and with GCP, and will compare the CRF entries with the source data. 
For source data verification, at least the following information must be included in the patient 
note/file: 
• Date of patient’s written informed consent, 
• Patient participation in a clinical study and the study number, 
• Patient visit dates, 
• Serious adverse events, 
• Adverse events, 
• Confirmation of the diagnosis of the indication being treated, 
• Study drug administration (e.g., start of study treatment, dose), 
• Concomitant disease medications and dose (including parenteral nutrition and 
corticosteroid dose), and 
• Primary endpoint. 
The CRA will collect the appropriate copy or copies of the completed forms. He/she will also 
verify the correct use of the investigational product. Entinostat will not be supplied to the 
investigator site prior to approval from the IRB/IEC and regulatory authority (if applicable). At a 
final visit, the CRA will check all remaining material including the remaining quantities of the 
investigational product and will organize its return. 
 

Entinostat and Aldesleukin 
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In addition, the CRA will determine whether all AEs and SAEs have been appropriately 
documented and whether the SAEs were reported within the time periods required. 
A member of the Quality Assurance Unit may arrange to visit the investigator in order to audit 
the performance of the study at the study site and the study documents originating there. A 
sponsor designee will usually accompany the auditors. The investigator will be informed about 
the outcome of the audit. 
 
11.7 Reporting and Exclusion 
 
11.7.1 Evaluation of toxicity 
 
 All patients will be evaluable for toxicity from the time of their first dose of entinostat 
 
11.7.2 Evaluation of response 
 
All conclusions should be based on all eligible patients who initiate entinostat  treatment. 
Sub-analyses may then be performed on the basis of a subset of  
patients, excluding 
those for whom major protocol deviations have been identified  (e.g., early death due to 
other reasons, early discontinuation of treatment, major  protocol violations, etc.).  
 
However, these sub-analyses may not serve as the basis for drawing conclusions 
concerning treatment efficacy, and the reasons for excluding patients from the analysis 
should be clearly reported. 
Prolonged lymphocyte decrease is not considered a DLT since it is a known side effect of 
high dose interleukin 2 and it is not considered clinically relevant. 
 
Enrolled patients who are deemed to be not eligible/not evaluable will be replaced. 
11.7.3 Discontinuation and Withdrawal of Subjects 
 
11.7.3.1 All patients who initiate protocol treatment will be included in the overall 
evaluation of response (intent-to-treat analysis). All reasons for discontinuation of 
treatment should be documented clearly in the medical record. Note: Patients who sign a 
consent form, but do not initiate study drug will be replaced. 
 
11.7.3.2 Non-compliance with the study protocol; including, but not limited to not 
 
attending the majority of scheduled visits. The PI will determine when  
 
 
non-compliance should lead to removal from study. Note: The patients  
 
 
will still be included in the overall evaluation of response (intent-to-treat  analysis 
11.7.3.3 Unacceptable major toxicity. Note: The patients will still be included in the 
overall evaluation of response (intent-to-treat analysis). 
11.7.3.4 At patient’s own request. Note: The reason for discontinuation from the  study 
must be documented. The patients will be included in the overall  evaluation of response 
(intent-to-treat analysis) if any protocol therapy was administered prior to withdrawal. 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
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12. ETHICS 
 
12.1 Institutional Review Board 
The study protocol and any amendment that is not solely of an administrative nature must be 
approved by an Independent Ethics Committee (IEC) or Institutional Review Board (IRB). 
 
12.2 
Evaluation of benefits and risks/discomforts 
 
Potential benefits: Patients will receive evaluation and treatment of their malignancy as a 
result of participating in this trial. The trial will provide information on how the combination of 
entinostat and aldesleukin should be administered to patients, but may or may not help a specific 
patient personally. This treatment may offer temporary control of the disease, but is not curative 
by this protocol. Because of the study design, patients may be treated with a drug dose that is too 
low to be effective. Therefore, neither benefit can be promised nor can the chance of benefit be 
predicted. Alternative approaches to entering this trial, including supportive care only, will also 
be discussed before the verbal and written consent is obtained regarding the risks, benefits, and 
the treatment requirements of this trial. 
Measures for minimizing risk: Administering entinostat and aldesleukin to patients may 
involve risks that are currently unforeseeable. Side effects can be unpredictable in nature and 
severity, although all care will be taken to minimize them. If patients suffer any physical injury 
as a result of participating in this study, immediate medical treatment is available at the treatment 
center. Frequent blood work will be taken to monitor side effects. Although no compensation is 
available, any injury will be evaluated and treated in keeping with the benefits or care to which 
patients are entitled under applicable regulations. Malignancies with no further standard 
treatment options generally have a poor prognosis. Therefore, patients may experience 
significant treatment-related morbidity, and/or complications from progression of their disease. 
12.3 
Risks/benefits analysis 
Data gathered from both clinical and laboratory evaluations in this trial will be analyzed 
frequently to ensure safety of patients. Any new or significant finding(s) found during the course 
of the research will be shared and explained to each participant since that may affect a patient’s 
willingness to participate further. Patient’s anonymity will be protected to the maximum extent 
in all publications and presentations that result from this research. 
12.4 
Patient information and consent 
The investigator will explain the nature of the study, its purpose and associated procedures, the 

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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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expected duration, and the potential benefits and risks of participation to each patient prior to 
his/her entry into the study (i.e., before examinations and procedures associated with selection 
for the study are performed). Each patient will have ample opportunity to ask questions and will 
be informed about the right to withdraw from the study at any time without any disadvantage and 
without having to provide reasons for this decision. Following this informative discussion, a 
patient will be asked if he/she is willing to sign and personally date a statement of informed 
consent. Only if the patient voluntarily agrees to sign the informed consent statement and has 
done so, may he/she enter the study. The patient will receive a copy of the signed and dated 
informed consent form. 
The signed informed consent statement is to remain in the investigator's files. The investigator 
will document on each CRF that he/she has informed the patient and that the patient has signed 
the informed consent statement. 
The informed consent form and any other written information provided to patients will be 
revised whenever important new information becomes available that may be relevant to the 
patient's consent, or there is an amendment to the protocol which necessitates a change to the 
content of the written informed consent form. The investigator will inform the patient of changes 
in a timely manner and will ask the patient to confirm continuation of his/her participation in the 
study by his/her signature on the revised informed consent form. Any revised written informed 
consent form must receive the IRB’s approval/favorable opinion in advance of use. 
 
12.5 
Data handling and record keeping 
12.5.1 Case report forms 
This study has been assigned to be monitored by the Clinical Trials Monitoring Service (CTMS). 
Theradex is the pharmaceutical company that administers the Clinical Trials Monitoring Service. 
CTMS Data is required to be submitted to CTMS at least every two weeks, via the NCI/DCTD 
case report forms or the electronic case report form (ACES). Details on the protocol specific 
requirements will be communicated to the principal investigator by Theradex. Adverse event 
data will also be collected in the electronic data entry system used by Roswell Park Clinical 
Research Services. Furthermore, the investigator will document in the patient files (hospital 
files). If corrections are necessary, they will be entered by an authorized member of the 
investigator’s staff in the following manner: the wrong entry will be crossed out with a single 
line so that it remains legible, and the correct entry will be placed next to it. All corrections will 
be initialed and dated by the site staff making the correction. For corrections concerning adverse 
events or the primary variable, a reason for any alteration must be provided. 

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Any documents related to the study must be archived at the study site or in a central archive. 
This includes the careful listing of the identities of the patients involved in the study. This list 
and the signed informed consent statements are key documents in the files to be stored by the 
investigator. 
Patient (hospital) files will be archived according to local regulations. All documents related to 
the study must be retained until at least 15 years after the end of the study. At the end of this 
period, CTEP will inform the investigator as to when these documents no longer need to be 
retained. 
 
12.5.2  Laboratory data and reference laboratories 
For laboratory services provided by the investigator, the investigator must supply the sponsor 
with a complete list of all normal laboratory values for the laboratory utilized, as well as proof of 
laboratory certification by the Clinical Laboratory Improvement Amendments. Shipping 
information for the pharmacodynamic samples is contained in section 7.6. 
12.5.3 Patient registry 
The investigator should maintain a patient registry of all patients entered into the study in the 
event a safety issue arises after study completion. 
12.5.4 Drug accountability 
The designated personnel will confirm receipt of the drugs investigational product in writing and 
will use the investigational product only within the framework of this clinical study and in 
accordance with this study protocol. He/she will keep a record of the investigational product 
dispensed, preferably on the CRF. The investigational product must be stored in a locked storage 
facility and protected from unauthorized access. Any unused, partially used, or empty containers 
of the investigational product will be returned to the sponsor at the termination of the study at the 
latest. Receipt, distribution, and return of the investigational product must be properly 
documented on the forms provided by the sponsor giving the following information: Study 
number, sender, receiver, data, mode of transport, quantity, batch number, and expire or retest 
date, if applicable. 
 
 
 

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13. REFERENCES 
 
1. 
Jemal, A, Tiwari RC, Murray T, et al. Cancer statistics, 2007. CA Cancer J Clin, 54: 8-
29, 2007. 
2. 
Yang JCY. Interleukin-2: Clinical Applications: Renal Cell Carcinoma (Rosenberg SA 
ed) In: Principles and Practice of the Biologic Therapy of Cancer. Third Edition 
Philadelphia: Lippincott Williams & Wilkins 2000:73-82. 
3. 
Negrier S, Escudier B, Lasset C et al. Recombinant human interleukin-2, recombinant 
human interferon alfa-2a, or both in metastatic renal-cell carcinoma. Groupe Francais 
d'Immunotherapie. N Engl J Med 1998;338(18):1272-8. 
4. 
Abrams JS, Rayner AA, Wiernik PH et al. High-dose recombinant interleukin-2 alone: a 
regimen with limited activity in the treatment of advanced renal cell carcinoma. JNCI 
1990;82(14):1202-6. 
5. 
Atkins MB, Dutcher J, Weiss G et al. Cytokine Working Group. Kidney cancer: the 
Cytokine Working Group experience (1986-2001): part I. IL-2-based clinical trials. 
Medical Oncology 2001;18(3):197-207. 
6. 
McDermott DF, Regan MM, Clark JI, Flaherty LE, Weiss GR, Logan TF, Kirkwood JM, 
Gordon MS, Sosman JA, Ernstoff MS, Tretter CP, Urba WJ, Smith JW, Margolin KA, 
Mier JW, Gollob JA, Dutcher JP, Atkins MB. Randomized phase III trial of high-dose 
interleukin-2 versus subcutaneous interleukin-2 and interferon in patients with metastatic 
renal cell carcinoma. J Clin Oncol. 2005 Jan 1;23(1):133-41. 
7. 
Yang JC, Sherry RM, Steinberg SM et al. Randomized study of high-dose and low-dose 
interleukin-2 in patients with metastatic renal cancer. J Clin Oncol 2003;21(16):3127-32. 
8. 
Escudier B, Eisen T, Stadler WM et al.: Sorafenib in advanced clear-cell renal-cell 
carcinoma. N Engl J Med. 2007 Jan 11;356(2):125-34 
9. 
Motzer RJ, Hutson TE , Tomczak p et al. Sunitinib versus interferon alfa in metastatic 
renal-cell carcinoma. N Engl J Med. 2007 Jan 11;356(2):115-24. 
10. 
Marks P, Rifkind RA, Richon VM, Breslow R, Miller T, Kelly WK. Histone deacetylases 
and cancer: causes and therapies. Nat Rev Cancer 2001; 1:194-202. 
11. 
Johnstone, R. W. Histone-deacetylase inhibitors: novel drugs for the treatment of cancer. 
Nat. Rev. Drug Discov., 1: 287-299, 2002 
12. 
Minucci, S. & Pelicci, P.G. Histone deacetylase inhibitors and the promise of epigenetic 
(and more) treatments for cancer. Nat. Rev. Cancer 6, 38–51 (2006). 
13. 
Saito A, Yamashita T, Mariko Y, et al. A synthetic inhibitor of histone deacetylase, MS-
27-275, with marked in vivo antitumor activity against human tumors. Proc. Natl. Acad. 
Sci. USA 1999; 96:4592-97. 
14. 
Ryan QC, Headlee D, Acharya M, et al. Phase I and pharmacokinetic study of entinostat, 
a histone deacetylase inhibitor, in patients with advanced and refractory solid tumors or 
lymphoma. J Clin Oncol 2005;23(17):3912-22. 
15. 
Wang XF, Qian DZ, Ren M et al. Epigenetic modulation of retinoic acid receptor beta2 
by the histone deacetylase inhibitor entinostat in human renal cell carcinoma. Clin Cancer 
Res 2005;11(9):3535-42. 
16. 
Kato Y, Yoshimura K, Shin T, Verheul H, Hammers H, Sanni TB, Salumbides BC, Van 
Erp K, Schulick R, Pili R. Synergistic in vivo antitumor effect of the histone deacetylase 
inhibitor entinostat in combination with interleukin 2 in a murine model of renal cell 
carcinoma. Clin Cancer Res. 2007 ;13:4538-46 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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17.  Acharya, M.R., J.E. Karp, E.A. Sausville, K. Hwang, Q. Ryan, I. Gojo, J. Venitz, W.D. 
Figg, and A. Sparreboom, Factors affecting the pharmacokinetic profile of entinostat, a 
novel histone deacetylase inhibitor, in patients with cancer. Invest New Drugs, 24(5): p. 
367-75. (2006). 
18. Simon, R. (1989). Optimal two-stage designs for phase II clinical trials. Cont. Clin. Trials 
10:1-10. 
 
 
 
 

Entinostat and Aldesleukin 
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I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 63 of 79 
 
 
APPENDIX A: Protocol for quantitative analysis of tumor metabolism and tumor blood 
flow using 18FDG–PET/CT imaging. 
 
1. Synopsis 
In order to assess the effect of entinostat and aldesleukin on tumor metabolism, measurements 
with PET/CT utilizing 18FDG will be performed in a standardized fashion. Studies will be 
undertaken in eligible patients participating in the clinical study. 
Patients will receive 7 mCi of 8FDG. Dynamic acquisition of 23 frames will be performed from 
0-60 minutes post injection. Full kinetic analysis will be undertaken using standard modeling 
techniques.  
PET/CT studies will be undertaken before initiating entinostat administration and 5 weeks after 
initiation of aldesleukin treatment. 
These studies will be conducted on patients enrolled by the Sydney Kimmel Comprehensive 
Cancer Center at Johns Hopkins University (JHU). Additional institutions may also participate as 
the study progresses.  
 
 
 
 
 
 
18FDG admin followed 
by ~60 min PET scan 
~60 min 
Transmission 
scan (10 min) 
~10 min 

Entinostat and Aldesleukin 
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I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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2. Glossary 
 
PET/CT 
Positron 
Emission 
Tomography/Computerized 
Tomography 
ROI 
Region of Interest 
SUV 
Standardized Uptake Value 
JHU 
Johns Hopkins University 
GE 
General Electric 
FDG 
Fluorodeoxyglucose 
T 1/2 
Half Life 
K1 
Model parameter representing influx of tracer in the ROI. 
 
3. Background and Rationale 
3.1. Tumor Metabolism 
Positron Emission Tomography (PET) is uniquely suited to evaluate metabolic activity in human 
neoplasms 
for 
diagnostic 
imaging 
purposes. 
The 
glucose 
analogue 
fluorine-18-
fluorodeoxyglucose (18FDG) has been proven successful as a PET imaging agent for detection 
and localization of many forms of cancer. The elevated rate of glycolysis in many types of tumor 
cells enhances the uptake of 18FDG in neoplasms relative to normal tissues. The most commonly 
used techniques for quantitative analysis are standardized uptake values (SUV) and simplified 
tracer kinetic modeling using Patlak-Gjedde analysis. Standardized uptake values are confined to 
the measurement of radioactivity concentrations at a fixed time point. 
18FDG PET/CT imaging may provide a useful pharmacodynamic tool in the assessment of tumor 
response upon immunotherapy in combination with HDAC inhibitors. 18FDG PET/CT imaging 
may also predict tumor response before anatomic assessment and help selecting patients who 
may benefit for further treatment with high dose aldesleukin. 
4. Patient Selection 
PET/CT scans will be performed in all eligible patients. 
Prior to any PET/CT study specific investigations being undertaken, freely given informed 
consent must be obtained from the patient. 
Only patients with suitable lesions ( approximately 1.0 cm) as measured by CT will be included 
in this sub study. Intrathoracic lesions are preferred, since the arterial input function is derived 
noninvasively from a large blood vessel (thoracic aorta). 
 
5. Procedures 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 65 of 79 
 
 
One scan per patient will be performed at each session. Each patient will undertake 2 separate 
sessions: before initiating entinostat administration and 5 weeks after initiation of aldesleukin 
treatment. 
At each session, one 18FDG PET/CT scan acquisition will be performed. Changes in metabolism 
from the baseline study will be computed. 
6. Tracer Preparation 
On the scheduled day of PET/CT assessments 18FDG will be produced at our cyclotron facility 
7. Scanning Protocol 
An eight-ring GE 4096 tomograph (GE Corp, Milwaukee, WI) will be used for scanning. The 
scanner has an axial resolution of 6.7 mm and in-plane resolution of 6.5 mm for studies 
conducted on patients enrolled by the Sydney Kimmel Comprehensive Cancer Center at Johns 
Hopkins. If additional centers are opened to undertake the sub study, every reasonable attempt 
will be made to standardize acquisition procedures to allowing pooling of data. 
The subject will be positioned in the tomograph in a supine position with the tumor region within 
the gantry and fixed on the bed by loose Velcro bands. Before emission scanning, a transmission 
scan for correction of photon attenuation will be performed for 10 minutes with a removable ring 
source containing 68 Ge.  
18FDG will be undertaken immediately follow with the patient not moving from the PET/CT 
scanner. In this manner, the same transmission data can be used for analysis of the emission 
18FDG. Subjects will receive 10 ml of 18FDG e i. v. (7 mCi) over 30 seconds using an automated 
injector. Dynamic acquisition of 23 frames will be performed as previously described; from 0-60 
minutes post injection. Full kinetic analysis will be available using standard modelling 
techniques. 
8. Safety 
Safety assessment and recording as described in the main protocol should be adhered to for 
patients enrolled in the sub study. 
9. Analysis 
9.1 General 
Analysis of PET/CT scan data will be undertaken at JHU using standard modelling techniques. 
Time-activity curves for each image slice are averaged over all image planes containing tumor to 
reduce noise in the resulting individual slice time-activity curves. 
Kinetic modelling will be performed in the 18FDG (three compartment) studies. SUV corrected 
for lean body mass is preferred. Region of interest (ROI) will be placed using comparator 
anatomic data (CT).  

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 66 of 79 
 
 
9.2 Metabolism 
Metabolism studies are performed after the intravenous injection of 18FDG. 
Relative changes in tumor metabolism (mg/gm of tissue/min) and SUV will be estimated. SUV 
for tumor may be regarded as a surrogate for metabolism.  
9.3 Interpretation of Estimated Parameters 
It is anticipated that with effective treatment there will be declines in the estimated tumor 
metabolic parameter. The precision of tumor SUV (semi quantitative metabolic rate) is 
reproducible within 10%. Further, SUV can be compared between institutions by comparing the 
SUV determined at JHU at varying times post 18FDG injection with the SUV determined at the 
same time point post injection. 
 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 67 of 79 
 
 
 
Reference 
 , Zasadny K, Helvie M, Hutchins GD, Weber B, Cody R. Metabolic monitoring of breast cancer 
chemohormonotherapy using positron emission tomography: initial evaluation. J Clin Oncol. 
1993 Nov;11(11):2101-11. 
 
APPENDIX B: Performance Status Criteria 
 
ECOG Performance Status Scale 
 
 
Karnofsky Performance Scale 
Grade 
Descriptions 
Percent 
Description 
0 
Normal activity. Fully active, able 
to 
carry 
on 
all 
pre-disease 
performance without restriction. 
100 
Normal, no complaints, no evidence 
of disease. 
90 
Able to carry on normal activity; 
minor signs or symptoms of disease. 
1 
Symptoms, 
but 
ambulatory. 
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). 
80 
Normal activity with effort; some 
signs or symptoms of disease. 
70 
Cares for self, unable to carry on 
normal activity or to do active work. 
2 
In 
bed 
<50% 
of 
the 
time. 
Ambulatory and capable of all 
self-care, but unable to carry out 
any work activities. Up and about 
more than 50% of waking hours. 
60 
Requires occasional assistance, but 
is able to care for most of his/her 
needs. 
50 
Requires considerable assistance and 
frequent medical care. 
3 
In bed >50% of the time. Capable 
of only limited self-care, confined 
to bed or chair more than 50% of 
waking hours. 
40 
Disabled, requires special care and 
assistance. 
30 
Severely disabled, hospitalization 
indicated. Death not imminent. 
4 
100% 
bedridden. 
Completely 
disabled. Cannot carry on any self-
care. Totally confined to bed or 
chair. 
20 
Very sick, hospitalization indicated. 
Death not imminent. 
10 
Moribund, 
fatal 
processes 
progressing rapidly. 
5 
Dead. 
0 
Dead. 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 68 of 79 
 
 
 
 
APPENDIX C: HDIL-2 (HIGH DOSE ALDESLEUKIN) ADMINISTRATION 
GUIDELINES 
The toxicities of IL-2 therapy are thought to result primarily from a capillary leak syndrome 
(CLS) as well as lymphoid infiltration which have been observed histologically in many organs. 
The earliest clinical manifestations of CLS are hypotension and tachycardia which can be seen 
two hours after the first dose of high-dose of IL-2. Decreased systemic vascular resistance has 
been measured at this time point and contributes to the early drop in blood pressure. Because of 
these changes, if patients are receiving antihypertensive medications they should be discontinued 
prior to initiating high-dose IL-2. 
Intravenous fluids are the initial therapy for hypotension and the goal is to maintain systolic 
blood pressure greater than 80-90 mmHg. It is common to require additional fluid in the first 24 
hours of therapy in response to hypotension and oliguria. After initial resuscitation, most patients 
reach a steady state of relative hypotension and tachycardia that will be maintained until the final 
doses of IL-2. No intervention is generally needed for these parameters unless they exceed 
predetermined guidelines or oliguria ensues. Fluid replacement of 1-1.5 liters/day above 
maintenance requirements should not be exceeded because of extravasation and compounding of 
generalized edema and pulmonary congestion. The use of additional fluids late in the course of 
treatment should be restricted because the transient benefit they achieve may be overshadowed 
by the unwanted consequences of fluid leak (special caution must be exercised if body weight is 
>10% above baseline). Crystalloid is favored over colloid for the treatment of hypotension based 
on a randomized study that showed both were equally effective. If hypotension persists despite 
judicious fluid resuscitation, vasopressor support with an agonist such as phenylephrine is 
indicated. The hypotensive effects of each IL-2 dose generally peak 4-6 hours after infusion and 
vasopressors are titrated accordingly. When phenylephrine can be weaned to <0.5 μg kg/min it is 
generally safe to proceed with additional IL-2 dosing. As immunotherapy nears completion, 
phenylephrine requirements increase; doses approaching 1.5-2.0 μg kg min and inability to wean 
by 8 hours suggest that IL-2 infusions should be discontinued. Blood pressure measurements 
generally return to baseline within 24-48 hours of discontinuation of IL-2. 
Within the first 8 hours of starting IL-2, decreased urine output is frequent and is a consequence 
of hypotension and decreased intravascular volume. Renal dysfunction during IL-2 has been 
described as prerenal in nature, transient and without evidence of intrinsic renal damage. 
Oliguria is first treated with fluid boluses; if 1-1.5 Liters of crystalloid does not restore urine 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 69 of 79 
 
 
flow, an indwelling urinary catheter is inserted and dopamine at renal perfusion doses (2 μg /kg/ 
min) is initiated. Urine output greater than 10-20 cc hour must be established before additional 
IL-2 dosing can be considered. Serum creatinine levels are measured at least daily and are 
considered when making decisions about IL-2 dosing. 
Patients receiving high-dose IL-2 may experience progressive shortness of breath during therapy 
which very infrequently requires endotracheal intubation or drainage of a pleural effusion. The 
mechanism of pulmonary congestion has been attributed to increased vascular permeability. 
Careful pretherapy screening of patients who are smokers or have large tumor burdens in the 
lung (FEV1 and forced vital capacity should be greater than 65% of predicted and PaO2 >75 on 
room air), advising smokers to quit 2 weeks before therapy as well as judicious fluid replacement 
during therapy have resulted in a low incidence of severe toxicity. The selective monitoring of 
transcutaneous O2 saturation has been very helpful in patients experiencing pulmonary 
symptoms. O2 saturation should be maintained above 95% and if this level cannot be met by 4L 
O2 by nasal cannula or 40% O2 by mask, IL-2 dosing should be discontinued. The auscultation 
of rales in the lung bases is not infrequent during therapy but progression to the mid lung fields 
coupled with marginal O2 saturation are reasons to discontinue IL-2 dosing. Chest radiographs 
revealing interstitial edema and pleural effusions provide complementary information to the 
clinical assessment and are performed selectively. After discontinuation of IL-2, symptoms of 
pulmonary congestion resolve promptly and recovery is aided by rapid diuresis. 
One of the more visible sequelae of CLS is the generalized total body edema and consequent 
weight gain during high-dose IL-2. Neurovascular compression from edema is rare and is best 
treated with elevation of the involved extremity and use of compression garments. Cautious use 
of fluids to correct hypotension and oliguria is important in minimizing edema. Diuresis is not 
always possible during therapy because patients may be hypotensive and are frequently 
unresponsive to diuretics. After stopping IL-2 and when the blood pressure is returning to 
baseline (and not requiring vasopressors), diuresis should be vigorously initiated by 
pharmacologic means. Fluid balance is rapidly restored and the majority of patients are 
discharged at or below admission weight about 3 days after stopping IL-2. 
Pre-Therapy Assessment and Interventions 
• Confirm no recent (< 7 days) history of infections (respiratory, renal, IV site, other). If 
present, delay IL-2 by 1 week. 
• Check labs (i.e. normalized since last IL-2), EKG (i.e. no changes since IL-2), CXR (i.e. 
no effusion since IL-2). 
• Draw research labs if appropriate. 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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• Start acetaminophen, indomethacin and H2 blocker 8-12 hours before IL-2. 
• Premedicate patient for nausea (before IL-2 dose) 
 
During Therapy Assessment and Interventions 
 
• Start maintenance IV at 100 cc hr before IL-2 is started. 
 
PATIENT MONITORING GUIDELINES 
 
 
  
 
 
 
Ward  
 
 
 
ICU 
 
 Vitals signs 
 
 
Every 4hr 
 
 
 
Every 1hr 
 
 Intake and output 
 
Every 4hr 
 
 
 
Every 1hr 
 
 Weight 
 
 
Daily  
 
 
 
Daily 
 
 Mental Status  
 
Every 8hr 
 
 
 
Every 8 hr 
 
 IV site  
 
 
Every 8hr (change 
 
 
Every 8 hr 
 
  
 
 
 
arm IV every 3 days) 
 
 Labs 
 
 CBC  
 
 
Daily  
 
 
 
BID 
 
 Lytes, BUN, Crt, Glu  
Daily  
 
 
 
BID 
 
 ALT, AST, total bili  
Daily  
 
 
 
Daily 
 
 Ca++, Mg++, phosphorus 
Daily  
 
 
 
Daily 
 
 PT, PTT 
 
 
Daily after day 2. 
 
 
Daily 
 
 CK, total 
 
 
Daily  
 
 
 
Daily 
 
 TSH, T3, T4  
 
Each cycle 
 
 
 
Each cycle 
 
 Urinalysis 
 
 
Each cycle 
 
 
 
Each cycle 
 
 EKG  
 
 
Each cycle 
 
 
 
Each cycle 
  
 CXR  
 
 
Each cycle 
 
 
 
Each cycle 
 
Target blood pressure during therapy 
 
Normal BP for patient 
Target BP on therapy 
<100 mm Hg  
 
>80 mm Hg 
100-120 mm Hg 
 
>85 mm Hg 
>120 mm Hg  
 
>90 mm Hg 
 
IL-2 dosing 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 71 of 79 
 
 
No dose reductions of IL-2 are performed. IL-2 doses are delayed according to symptomatic 
recovery from the previous dose. Delay greater than 24 hours should result in discontinuation of 
a course of IL-2. 
Guidelines for delay or discontinuation of IL-2 are given below. The presence of relative criteria 
implies that a patient is nearing the completion of a course of therapy but that with appropriate 
corrective measures or with a time delay to allow for recovery, it may be safe to proceed. Several 
relative criteria are usually an indication to discontinue dosing. The presence of absolute criteria 
is generally considered an indication to stop dosing IL-2. 
 
NO steroids, systemic or topical, should be used at any time while patients are receiving 
immunotherapy. 
 
DOSE MODIFICATION FOR HDIL-2 
 
(Delay or discontinuation only; no dosage adjustment) 
 
System 
 
Relative Criteria 
 
Absolute Criteria 
 
Cardiac 
 
Sinus Tach 120-130  
Sinus Tach >130 
(persists after correcting hypotension, fever, 
stopping dopamine) 
 
  
 
 
 
 
 
EKG changes of ischemia 
 
  
 
 
 
 
 
Atrial fibrillation 
 
  
 
 
 
 
 
Supraventricular tachycardia 
 
  
 
 
 
 
 
Ventricular arrhythmias  
(frequent PVCs, bigeminy, v-tach) 
 
  
 
 
 
 
 
Elevated CPK-MB 
 
Dermatologic  
 
 
 
 
Moist desquamation 
 
Gastrointestinal 
Diarrhea, 1000 cc/shift 
Diarrhea, 1000 cc/shift x 2 
Vomiting not responsive to medication 
 
 lleus/abdominal distention 
Severe 
abdominal 
distention 
affecting 
breathing 
 
  
 
 
 
 
 
Severe abdominal pain, unrelenting 
Hemodynamic  
Max Neo-Synephrine  
Max Neo-Synephrine 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 72 of 79 
 
 
 
  
 
1-1.5 ug/kg/min 
 
1.5-2.0 ug/kg/min 
 
  
 
Min Neo-Synephrine  
Min Neo-Synephrine 
 
  
 
>0.5 ug/ kg/ min 
 
> 0 8 ug/ kg/ min 
Hemorrhagic  
Guaiac+ sputum, emesis, 
Frank blood in sputum, emesis, stool 
 
  
 
Platelets 30-50,000/mm3 
Platelets <30,000/mm3 
 
Infectious 
 
 
 
 
 
Strong clinical suspicion or documented 
 
Neurologic 
 
Vivid dreams  
 
Mental status not reversible in 2 hrs. 
 
  
 
Emotional lability 
 
Disorientation 
 
  
 
 
 
 
 
Hallucinations 
 
Pulmonary 
 
Resting shortness of breath 
>4 L 02 by NC for sat >95% 
 
  
 
3-4 L 02 NC for sat >95% 
40% 02 mask for sat > 95% 
 
  
  
 
 
 
 
Endotracheal Intubation 
 
  
 
Rales 1/3 up chest 
  
Rales 1/2 up chest 
Pleural effusion requiring tap or chest tube 
while on therapy 
 
Renal  
 
Urine 80-160 cc/8 hour 
Urine <80 cc/8 hour 
 
  
 
Urine 10-20 cc/hr 
 
Urine <10 cc/ hr 
 
  
 
Creatinine 2.5-2.9 mg/dl 
Creatinine >3.0 mg/dl 
 
Weight Gain  
15% weight gain over baseline 
 
INTERVENTION 
 
Observation category Action 
 
≤ 3 Relative criteria Initiate corrective measures 
>3 Relative criteria 
Stop IL-2 for current course 
1 Absolute criterion Stop IL-2 for current course 
 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 73 of 79 
 
 
 
CORRECTIVE MEASURES 
Symptom/sign 
Corrective measure 
Arrhythmia (other than 
sinus tach) 
Stop IL-2 (most arrhythmias). Correct electrolytes, minerals, anemia, 
hypoxia; use medications as indicated 
Anemia 
Transfuse PRBCs to achieve Hct > 28 % during IL-2 dosing. 
Acidosis 
Total C02 (bicarbonate) < 20 mmol/ L, give 50 meq bicarb IV 
If CO2<18 mmol/L, give 100 meq bicarb IV 
Chills (generally after 
first or second dose) 
Warm blankets as first measure. 
IV Demerol if persist. 
 
CK elevation 
Measure isoenzymes, EKG. If have evidence of myocarditis, must 
stop IL-2. 
Will need a MUGA stress test before next cycle of IL-2 to rule out 
Myocardial dysfunction. 
Future IL-2 may be considered if the MUGA is normal. 
 
Dermatitis 
Oatmeal baths, lotions (NO steroid or alcohol containing lotions). 
Diarrhea 
Antidiarrheals (alternate medications). Avoid overuse because of 
complicating ileus and distention. 
Edema 
Elevate symptomatic extremity. Use fluids judiciously. 
Epigastric pain 
Evaluate cause, give indomethacin rectally, and consider antacids. 
Fever breakthrough 
Increase frequency of indomethacin to every 6 hr. 
Consider septic work-up if happens after first 24 hrs. of therapy i.e. 
high spike, 38.5 or above, during therapy. 
Hypoalbuminemia 
Observe 
Hypocalcemia 
Maintain above lowest normal at institution (compensate for low 
albumin). 
 
Hypokalemia 
Maintain potassium above 3.6 mmol/L. 
Hypomagnesemia 
Maintain above lowest normal at institution (compensate for low 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 74 of 79 
 
 
albumin) 
Hypotension 
Use crystalloids; maintain systolic BP > 80-90 mm Hg. 
If hypotension persisting in spite of optimal fluid therapy consider 
vasopressors. See under dose modification for hemodynamic criteria 
Infection 
Stop IL-2 and treat infection as indicated 
Mucositis/stomatitis 
Frequent oral care, mouthwashes, topical anesthetics, nystatin. 
Oliguria 
Fluid boluses at first. If 1-1.5 L of crystalloid does not restore U/O, 
insert Foley catheter and initiate renal dose dopamine (2 ug/kg/min). 
U/O should be 20-30 cc hour at least. 
Nausea vomiting 
Antiemetics (alternate medications and routes if not effective). 
Nasal congestion 
Room humidifier, decongestant (NO topical steroids). 
Poor venous access 
Insert PIC or central line. Antibiotic 
 
Prophylaxis started with PIC or central line inserted and continued 
until line out (remove line as soon as not needed for fluids, 
vasopressors, replacements, diuretics, etc). 
 
Prophylaxis started with PIC or prophylaxis started with PIC or 
central line inserted and continued until line out (remove line as soon 
as not needed for fluids, vasopressors, replacements, diuretics, etc). 
 
 
Pruritus 
 
 
Prophylaxis started with PIC or central line inserted and continued 
until line out (remove line as soon as not needed for fluids, 
vasopressors, replacement, diuretics) 
Oatmeal baths, lotions, antipruritics 
 
Shortness of breath 
Check transcutaneous 02 sat. If <95% use 02. Use fluids judiciously. 
Do not use inhalational steroids 
 
Tachycardia (sinus) 
Correct 
fever, 
hypotension, 
hypoxia, 
anemia, 
consider 
discontinuation of dopamine if used. 
 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 75 of 79 
 
 
Thrombocytopenia 
Consider transfusion if count < 20,000/mm3 
 
Concomitant Medications Used During IL-2 Therapy* 
 
Medication 
 
 
 
Dose/Frequency/Route  
 
Side Effect Treated 
Acetaminophen  
 
 
650 mg q4h, PO/PR  
 
 
Fever, myalgia 
Indomethacin  
 
 
50-75 mg q8h, PO/PR   
 
Fever, myalgia 
Meperidine HCL 
 
 
25-50 mg q1h, IV prn   
 
Chills 
Ranitidine HCL  
 
 
50 mg q8h, IV   
 
 
Gastritis 
Aluminum hydroxide (200 mg)  
30 ml q3h, PO prn  
 
 
Gastric upset 
Magnesium hydroxide 200 mg 
Simethicone 20 mg 
Droperidol 
 
 
 
1 mg q4-6h, IV prn  
 
 
Nausea 
Prochlorperazine 
 
 
25 mg q4h, PR prn or 10 mg  
 
Nausea 
q6, IV prn 
Ondansetron HCL 
 
 
10 mg q8h, IV prn  
 
 
Nausea 
Granisetron HCL 
 
 
0.01 mg/kg daily, prn   
 
Nausea 
Lorazepam 
 
 
 
0.5 – 1.0 mg q6h, PO/IV prn  
 
Nausea, anxiety 
Haloperidol 
 
 
 
1-5 mg q1h, IV/IM prn   
 
Agitation 
Loperamide 
 
 
 
2 mg q3h, PO prn 
 
 
Diarrhea 
Diphenoxylate HCL 
 
 
(2.5 mg) q3h, PO prn   
 
Diarrhea 
Atropine sulfate (25 µg) 
Codeine Sulfate  
 
 
30 – 60 mg q4H, PO prn  
 
Diarrhea 
Tucks  
 
 
 
Apply locally, prn  
 
 
Perianal discomfort 
Hydroxyzine HCL 
 
 
10 – 20 mg q6h, PO prn  
 
Pruritis 
Diphenhydramine HCL  
 
25 – 50 mg q4h, PO/IV prn  
 
Pruritis 
Oatmeal powder/baths  
 
Apply locally prn 
 
 
Pruritis 
Lubriderm 8 oz. with 0.25%  
 
Apply locally prn  
 
 
Pruritis 
Camphor and 0.25% menthol 
Sodium bicarbonate 
 
 
6 tsp/1500 ml Swish & swallow prn 
Mucositis 
Lidobenalox oral 
 
 
5 ml q3h, PO prn  
 
 
Mucositis 
Pseudoephedrine HCL  
 
30 mg q6h, PO prn  
 
 
Sinus congestion 
Temazepan 
 
 
 
15-30 mg qhs, PO prn   
 
Insomnia 
Zolpidem 
 
 
 
5-10 mg qhs, PO prn  
 
 
Insomnia 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
page 76 of 79 
 
 
Normal saline  
 
 
250-500 ml IV prn  
 
 
Oliguria, hypotension 
Dopamine HCL  
 
 
400 mg/250 ml, IV prn   
 
Oliguria 
(2 µg/kg/min) 
 
Phenylephrine  
 
 
40 mg/100 ml, titrate IV prn  
 
Hypotension 
(0.1 – 2.0 µg/kg/min) 
Calcium gluconate 10%  
 
1 gm (over 1h), IV prn   
 
Hypocalcemia 
Magnesium sulfate 
 
 
1 gm (over 1h), IV prn   
 
Hypomagnesemia 
Potassium phosphate 
 
 
10 – 15 mmole (over 6h), IV prn  
Hypophosphatemia 
Potassium chloride 
 
 
10 meq (over 1h), IV prn  
 
Hypokalemia 
Cefazolin 
 
 
 
1-2 gm q6h, IV   
 
 
Prevent line sepsis 
Clindamycin 
 
 
 
900 mg q8h, IV  
 
 
Prevent line sepsis 
 
Abbreviations: q: every; PO: orally; PR: rectally; IV: intravenously; PRN: as needed. 
* Douglas Schwartzentruber, Biologic Therapy of Cancer, editors: VT De Vita, Jr, S Hellman 
and SA Rosenberg, 2nd edition, JB Lippincott, 1995, pg. 235-249. 
 
 

Entinostat and Aldesleukin 
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NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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Appendix D: Instructions for Network Sites 
 
1. CONTACT INFORMATION 
All questions related to the protocol or study implementation should be directed to: 
Roswell Park Cancer Institute 
CRS Quality Assurance Office 
GBSB 1930 
Buffalo, New York 14263 
Telephone: 
716-845-8084 or 716-845-1205 - M-F; 8:00 AM to 5:30 PM  
716-845-2300 - After hours, Weekends and Holidays: request the RPCI Principal 
Investigator 
Fax: 716-845-8743 
 
2. INFORMED CONSENT 
▪ Informed Consent must be obtained by the Investigator from any patients wishing to 
participate, prior to any procedures or change in their treatment 
▪ An informed consent template is provided by RPCI and can be amended to reflect 
institutional requirements 
▪ All consent changes must be reviewed by Roswell Park Cancer Institute Quality 
Assurance Office prior to submission to the site IRB. 
▪ The informed consent must be IRB approved 
▪ Always check that you are using the correct date and version of the IRB approved 
consent. 
 
3. PATIENT REGISTRATION  
The Subject Enrollment Log must be faxed to the CRS Quality Assurance Office within 24 
hours of the date the patient is consented. Once the Principal Investigator has determined that 
the eligibility criteria have been met, complete the Patient Registration Form and fax it to 
the RPCI Quality Assurance Coordinator at (716) 845-8743. 
Note: The patient completes the Gender, Race, and Ethnicity form and this is placed in the 
study binder. 
Roswell Park Cancer Institute does not grant exceptions to eligibility criteria. 
 
4. STUDY DEVIATIONS 
▪ If a deviation has occurred to eliminate hazard, this should be reported to the RPCI 
Quality Assurance Office, site IRB and any other regulatory authority involved in the 
trial. 
▪ ANY study deviation will be recorded on the Study Deviation Log 
▪ Patients who are inadvertently enrolled, with significant deviation(s) from the study- 
specified criteria, will be removed from the study 
▪ Notify RPCI of any early patient withdrawal and appropriately document the 
discontinuation and the reason why. 

Entinostat and Aldesleukin 
PI: Saby George, M.D 
I 145208 
NCI Protocol #: 7870  
Protocol Version Date  5/29/19 
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5. STUDY DOCUMENTATION 
▪ Study documents must be filled out completely and correctly. Ditto marks are not 
allowed. 
▪ If an entry has been documented in error put a single line through the entry and initial and 
date the change. The auditor must be able to read what has been deleted.  
o 
Do NOT use white-out, magic marker, scratch-outs 
o 
Do NOT erase entries  
▪ Use only black ink for documentation on the accountability form and any other study 
forms. 
 
6. DRUG ACCOUNTABILITY 
Drug accountability will be strictly maintained, recording quantities of study drug 
received, dispensed to patients and wasted, lot number, date dispensed, patient ID 
number and initials, quantity returned, balance remaining, manufacturer, expiration 
date, and the initials of the person dispensing the medication.  
▪ Responsibility rests solely with the Principal Investigator but can be delegated as 
appropriate (e.g. to pharmacy personnel) 
▪ Records must be maintained regarding receipt, dispensing, return, waste and disposition 
of all investigational agents 
▪ Study drug supply should only be used in accordance with the IRB approved study 
▪ Drug accountability forms are protocol and agent specific, they are study source 
documents and will be used to verify compliance with the study 
▪ Any discrepancies shall be documented and explained 
▪ An inventory count should be performed with each transaction 
▪ Drug accountability forms shall be stored with study related documents 
▪ Each medication provided for this study and each dosage form and strength must have its 
own Drug accountability.  
▪ Do NOT “transfer”, “borrow” or “replace” supplies between studies 
▪ Dispensing the wrong study supply is considered a medication error 
▪ Never replace investigational agents with commercial product 
 
7. SERIOUS ADVERSE EVENT REPORTING: 
The site Investigator or designated research personnel will report all serious adverse events, 
whether related or unrelated to the study drug(s) to the IRB in accordance with their local 
institutional guidelines. Expedited adverse event (AE) reporting for this study is via CTEP-
AERS(CTEP Adverse Event Reporting System), accessed via the secure CTEP web site ( 
https://eapps-ctep.nci.nih.gov/ctepaers/pages/task?rand). The reporting procedures to be followed 
are presented in the “CTEP, NCI Guidelines: Expedited Adverse Event Reporting Requirements” 
which 
can 
be 
downloaded 
from 
the 
CTEP 
web 
site 
(http://ctep.cancer.gov/ 
protocolDevelopment/elcetronicapplications/docs/aeguidelines.pdf.). The site will notify the 
CRS Quality Assurance Office within one business day of being made aware of the SAE by 
faxing a copy of the expedited reports submitted to CTEP via CTEP-AERS. A preliminary 
written report must follow within 24 hours (1 business day) of the oral notification. A complete 
follow-up report must be filed within 10 working days. 
 
 

Entinostat and Aldesleukin 
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8. UNANTICIPATED PROBLEM REPORTING: 
An Unanticipated Problem (UAP) is any incident, experience, or outcome that meets all of 
the following criteria: 
 
1. Unexpected (in terms of nature, severity, or frequency) given: 
(a) the research procedures that are described in the study- related documents, 
including study deviations, as well as issues related to compromise of patient 
privacy or confidentiality of data;  
(b) the characteristics of the subject population being studied; 
2. 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); 
3. Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized; 
 
For all adverse events occurring that are unanticipated and related or possibly related to the 
research drug, biologic or intervention the participating physician or delegated research staff 
from each site will notify their local IRB in accordance with their local institutional 
guidelines. The site must also notify the CRS Quality Assurance Office within 24 hours of 
being made aware of the Unanticipated Problem by completing the RPCI Unanticipated 
Problem Report Form and faxing it to the CRS Quality Assurance office.