Setting Endotoxin Limits During Development of Investigational Oncology Drugs and Biological Products Guidance for Industry: Draft Guidance for Industry

Administrative draft guidance, not enacted legislation. This captured publisher edition preserves the listed document and attachments; later developments are not independently consolidated.

Endotoxin Draft Guidance
Setting Endotoxin Limits
During Development of
Investigational Oncology Drugs
and Biological Products
Guidance for Industry
DRAFT GUIDANCE
This guidance document is being distributed for comment purposes only.
Comments and suggestions regarding this draft document should be submitted within 60 days of
publication in the Federal Register of the notice announcing the availability of the draft
guidance. Submit electronic comments to https://www.regulations.gov. Submit written
comments to the Dockets Management Staff (HFA-305), Food and Drug Administration, 5630
Fishers Lane, Rm. 1061, Rockville, MD 20852. All comments should be identified with the
docket number listed in the notice of availability that publishes in the Federal Register.
For questions regarding this draft document, contact (CDER) Patricia Keegan at 301-796-1387,
or (CBER) Office of Communication, Outreach and Development at 800-835-4709 or 240-402-
8010.
U.S. Department of Health and Human Services
Food and Drug Administration
Oncology Center of Excellence (OCE)
Center for Drug Evaluation and Research (CDER)
Center for Biologics Evaluation and Research (CBER)
July 2020
Clinical/Medical

Setting Endotoxin Limits
During Development of
Investigational Oncology Drugs
and Biological Products
Guidance for Industry
Additional copies are available from:
Office of Communications, Division of Drug Information
Center for Drug Evaluation and Research
Food and Drug Administration
10001 New Hampshire Ave., Hillandale Bldg., 4th Floor
Silver Spring, MD 20993-0002
Phone: 855-543-3784 or 301-796-3400; Fax: 301-431-6353
Email: druginfo@fda.hhs.gov
https://www.fda.gov/drugs/guidance-compliance-regulatory-information/guidances-drugs
and/or
Office of Communication, Outreach, and Development
Center for Biologics Evaluation and Research
Food and Drug Administration
10903 New Hampshire Ave., Bldg. 71, Rm. 3128
Silver Spring, MD 20993-0002
Phone: 800-835-4709 or 240-402-8010
Email: ocod@fda.hhs.gov
https://www.fda.gov/vaccines-blood-biologics/guidance-compliance-regulatory-information-biologics/biologics-guidances
U.S. Department of Health and Human Services
Food and Drug Administration
Oncology Center of Excellence (OCE)
Center for Drug Evaluation and Research (CDER)
Center for Biologics Evaluation and Research (CBER)
July 2020
Clinical/Medical

Contains Nonbinding Recommendations
Draft — Not for Implementation
TABLE OF CONTENTS
I. 	INTRODUCTION............................................................................................................. 1
II. 	BACKGROUND ............................................................................................................... 2
III. 	ENDOTOXIN CONTROL AND LIMITS...................................................................... 2
IV. 	SETTING ENDOTOXIN LIMITS DURING DRUG DEVELOPMENT ................... 4
A. 	Early Clinical Development .......................................................................................................... 4
1. Parenteral (Excluding Intrathecal or Intraocular) Route of Administration .................................. 4
2. Intrathecal Route of Administration ................................................................................................ 5
B. 	Late Stage Clinical Development .................................................................................................. 5
REFERENCES.............................................................................................................................. 6

Contains Nonbinding Recommendations
Draft — Not for Implementation
1
Setting Endotoxin Limits During Development of Investigational	1
Oncology Drugs and Biological Products	2
Guidance for Industry1
3
4
5
6
This draft guidance, when finalized, will represent the current thinking of the Food and Drug	7
Administration (FDA or Agency) on this topic. It does not establish any rights for any person and is not	8
binding on FDA or the public. You can use an alternative approach if it satisfies the requirements of the	9
applicable statutes and regulations. To discuss an alternative approach, contact the FDA staff responsible	10
for this guidance as listed on the title page.	11
12
13
14
I. 	INTRODUCTION	15
16
New approvals in oncology often build on prior success by adding new drugs to current regimens	17
or by combining products in a novel treatment regimen, creating new multidrug regimens2 that	18
may have greater efficacy.3	
19
20
This guidance describes FDA’s recommendations to investigational new drug sponsors for	21
setting endotoxin limits during the development of investigational drugs intended for use in	22
combination with other approved drugs or for the codevelopment of two or more investigational	23
drugs. The scope of this guidance is limited to anticancer drugs, including combination products	24
under 21 CFR Part 3,4 as described further in this guidance and administered parenterally (except	25
for intraocular administration) to treat serious and life-threatening cancers based on histology or	26
stage of disease. This guidance does not apply to the development of drugs for adjuvant or	27
neoadjuvant treatment or for cancer subtypes that can be cured or where prolonged survival can	28
be achieved with available therapy.	29
30
In general, FDA’s guidance documents do not establish legally enforceable responsibilities.	31
Instead, guidances describe the Agency’s current thinking on a topic and should be viewed only	32
as recommendations, unless specific regulatory or statutory requirements are cited. The use of	33
1 This guidance has been prepared by the Oncology Center of Excellence in cooperation with the Center for Drug
Evaluation and Research and the Center for Biologics Evaluation and Research at the Food and Drug
Administration.
2 For the purpose of this guidance, regimen refers to two or more therapeutic products that are (or will be) marketed
separately but are being tested for use in combination based upon one or more adequate and well-controlled trials.
3 See the guidance for industry Codevelopment of Two or More New Investigational Drugs for Use in Combination
(June 2013). We update guidances periodically. To make sure you have the most recent version of a guidance,
check the FDA guidance web page at https://www.fda.gov/regulatory-information/search-fda-guidance-documents.
4 Additional considerations regarding endotoxin exposure may apply for delivery devices and combination products
with device constituent parts under 21 CFR 812.

Contains Nonbinding Recommendations
Draft — Not for Implementation
2
the word should in Agency guidances means that something is suggested or recommended but	34
not required.	35
36
37
II. 	BACKGROUND	38
39
Investigational drugs for treating patients with incurable cancers who have short life expectancy	40
are often studied in combination with approved drugs in an attempt to identify multidrug or	41
multimodality regimens that may prolong survival. Administration of one or more drugs within	42
the same 60-minute time frame as the investigational drug may pose challenges regarding the	43
endotoxin limits for that investigational drug at a time when the manufacturing process for that	44
investigational drug has not been optimized. This guidance addresses endotoxin limits for	45
investigational drugs for the treatment of advanced cancer, when evaluated in early (pilot)	46
clinical trials as part of a multidrug regimen.	47
48
FDA recognizes that based on the potential benefits of investigational drugs administered to	49
patients with life-threatening incurable cancers, such patients’ possible exposure to increased	50
levels of endotoxin exceeding the recommendations in USP General Chapter <85> Bacterial	51
Endotoxins Test may be considered an acceptable risk under appropriate circumstances.	52
53
This guidance discusses FDA’s thinking regarding setting endotoxin limits for investigational	54
drugs during early product development for incurable cancers. The recommendations are	55
consistent with the abbreviated product testing for feasibility clinical trials of monoclonal	56
antibodies in patients with serious or immediately life-threatening conditions for which no	57
effective alternative treatment exists.5 In this guidance, FDA recommends a risk-based approach,	58
weighing the potential risks of possible exposure to increased levels of endotoxin across all	59
components of a multidrug regimen against the potential benefits to patients with serious and	60
life-threatening cancers.	61
62
63
III. 	ENDOTOXIN CONTROL AND LIMITS	64
65
Bacterial endotoxins are lipopolysaccharides found in the outer membrane of gram-negative	66
bacteria. Endotoxins are released upon cell death and lysis and have the potential to contaminate	67
drug and biological products. When administered in high amounts, endotoxins can cause	68
pyrogenic reactions, severe inflammatory responses, septic shock, and death. Therefore, it is	69
important to prevent the introduction of bacterial endotoxins into drug and biological products	70
and their components (as defined in 21 CFR 201.3) during the product and component	71
manufacturing processes, and to conduct appropriate testing for the presence of endotoxins on	72
each batch of clinical and commercial product. Following are several guidances that refer to the	73
need for pyrogen and endotoxin control and testing during sterile drug and biological product	74
development:	75
76
5 See Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use
(February 1997).

Contains Nonbinding Recommendations
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• 	Guidance for industry Content and Format of Investigational New Drug Applications	77
(INDs) for Phase 1 Studies of Drugs, Including Well-Characterized, Therapeutic,	78
Biotechnology-derived Products (November 1995)	79
80
• 	Guidance for industry INDs for Phase 2 and Phase 3 Studies–Chemistry, Manufacturing,	81
and Controls Information (May 2003)	82
83
• 	Guidance for reviewers and sponsors Content and Review of Chemistry, Manufacturing,	84
and Control (CMC) Information for Human Somatic Cell Therapy Investigational New	85
Drug Applications (INDs) (April 2008)	86
87
• 	Guidance for industry Chemistry, Manufacturing, and Control (CMC) Information for	88
Human Gene Therapy Investigational New Drug Applications (INDs) (January 2020)	89
90
• 	Guidance for industry CGMP for Phase 1 Investigational Drugs (July 2008)	91
92
93
Controls on pyrogen and endotoxins are part of the chemistry, manufacturing, and control	94
information to be included in an investigational new drug, a new drug, or a biologics license	95
application for most drug products (21 CFR 312.23(a)(7)(iv)(b), 21 CFR 314.50(d)(1)(ii)(a), and	96
21 CFR 601.2(a) and (c), respectively). Similarly, current good manufacturing practice	97
regulations also require control of pyrogen for containers and closures (see 21 CFR 211.94(c)	98
and (d) and 21 CFR 600.11(h) and for most drugs (see 21 CFR 211.165(a), 21 CFR 211.167(a),	99
and 21 CFR 610.13(b)). As described in the guidance for industry Pyrogen and Endotoxins	100
Testing: Questions and Answers (June 2012), the requirement in 21 CFR 610.13 for rabbit	101
pyrogen testing for certain biological products may be waived if a method equivalent to the	102
pyrogen test is demonstrated in accordance with 21 CFR 610.9.6	
103
104
Controls on pyrogen and endotoxins are part of the chemistry, manufacturing, and control	105
information to be included in an investigational new drug, a new drug, or a biologics license	106
application (see 21 CFR 312.23(a)(7)(iv)(b), 21 CFR 314.50(d)(1)(ii)(a), and 21 CFR 601.2(a)	107
and (c), respectively). Similarly, current good manufacturing practice regulations also require	108
control of pyrogen (see 21 CFR 211.94(c)(d), 21 CFR 211.165(a), 21 CFR 211.167(a), 21 CFR	109
600.11(h), 21 CFR 610.9 (equivalent methods and processes), and 21 CFR 610.13(b)).	110
111
The compendial recommendations for threshold pyrogenic doses of endotoxins for injectable	112
products are based upon the results of human and animal studies demonstrating the deleterious	113
effects of bacterial endotoxins administration. United States Pharmacopeia (USP) General	114
Chapter <151> Pyrogen Test provides a test method and acceptance criteria for the absence of	115
pyrogens by measuring febrile reactions to injectable products. The endotoxin limits for	116
parenteral drugs recommended in USP General Chapter <85> Bacterial Endotoxins Test are	117
defined by the formula K/M, where K is the threshold pyrogenic dose of endotoxins per kg of	118
6 FDA supports the principles of the 3Rs, to reduce, refine, and replace animal use in testing when feasible. We
encourage sponsors to consult with us if they wish to use a nonanimal testing method they believe is suitable,
adequate, validated, and feasible. We will consider if the alternative method could be assessed for equivalency to an
animal test method.

Contains Nonbinding Recommendations
Draft — Not for Implementation
4
body weight or m2 of body surface area per hour and M is the maximum dose to be administered	119
to the patient within a 60-minute period. K is equal to 5 EU (endotoxin units) per kg of body	120
weight or 100 EU per m2 of body surface area per hour for nonintrathecal injections. K is equal	121
to 0.2 EU per kg of body weight per hour for intrathecal injections. These compendial	122
recommendations represent the upper limit for endotoxin content for injectable products so	123
stricter limits may be warranted.	124
125
These values represent the maximum recommended exposure to endotoxins, based on the	126
absence of clinically important increases in body temperature at these exposures. FDA expects	127
that sponsors of investigational new drug applications will justify the proposed endotoxin	128
acceptance criteria for each investigational therapeutic biologic, drug, or combination therapy	129
based on manufacturing experience and established control strategies, such as careful selection	130
of ancillary materials, aseptic processing, and the use of closed systems.	131
132
133
IV. 	SETTING ENDOTOXIN LIMITS DURING DRUG DEVELOPMENT	134
135
In keeping with the principles of facilitating drug development for serious and life-threatening	136
diseases, this guidance outlines FDA’s current thinking on a risk-based approach to setting	137
acceptance criteria for endotoxins during the clinical development of drugs intended to treat	138
serious and life-threatening cancers. Additionally, sponsors should refer to recommendations	139
regarding testing discussed in the guidance for industry Pyrogen and Endotoxins Testing:	140
Questions and Answers.	141
142
A. 	Early Clinical Development	143
144
During early clinical trials (e.g., dose-finding or activity-estimating trials) conducted in patients	145
with a serious and life-threatening cancer based on stage of disease and expected prognosis for	146
that cancer subtype:	147
148
1. 	Parenteral (Excluding Intrathecal or Intraocular) Route of Administration	149
150
• 	For investigational products that are small molecules or certain therapeutic biological	151
products,7 the endotoxin limits of an investigational drug or the combined endotoxin	152
limits of multiple investigational drugs administered concomitantly by a parenteral route	153
should not exceed the limit specified in USP General Chapter <85>, that is, 5 USP-EU	154
per kg body weight per hour or 100 EU per m2 body surface area (BSA) per hour. The	155
sponsor of an investigational new drug need not consider the potential endotoxin	156
contribution from approved and/or licensed components of a combination regimen when	157
calculating the acceptable limits for endotoxin exposure from the investigational drug.	158
159
• 	For all other investigational products, including cellular therapy and gene therapy	160
products, in order to assess causality of observed adverse events, the combined endotoxin	161
exposure from all agents (investigational drug and other approved and/or licensed	162
7 See Transfer of Therapeutic Products to the Center for Drug Evaluation and Research (CDER) at
https://www.fda.gov/AboutFDA/CentersOffices/OfficeofMedicalProductsandTobacco/CBER/ucm133463.htm.

Contains Nonbinding Recommendations
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concomitantly administered drugs) administered parenterally should not exceed the limits	163
specified in USP General Chapter <85>; this will allow better identification of adverse	164
reactions of investigational products that may overlap with the onset of and mimic the	165
signs and symptoms of endotoxin exposure.	166
167
2. 	Intrathecal Route of Administration	168
169
When investigational products will be administered intrathecally with other agents, the combined	170
endotoxin exposure from all drugs (investigational drug and other concomitantly administered	171
investigational, approved, or licensed drugs) should not exceed that specified in USP General	172
Chapter <85>, that is, 0.2 USP-EU per kg body weight per hour.	173
174
• 	In the rare case that the combined endotoxin exposure exceeds the limits described above,	175
sponsors should justify that such limits cannot be achieved based on specific aspects of	176
product manufacturing and provide a rationale to support a conclusion that the risks to	177
human subjects are reasonable considering the preliminary evidence of clinical activity of	178
the investigational product, the seriousness of the disease, and the availability of	179
satisfactory alternative therapies.	180
181
182
B. 	Late Stage Clinical Development	183
184
During clinical trials intended to support the approval of an investigational drug to be	185
administered concomitantly with other drugs:	186
187
• 	Sponsors should tighten the specifications for endotoxin limits to ensure that by the time	188
they submit a marketing application for that drug, the endotoxin limits will not exceed	189
that specified in USP General Chapter <85> for a parenterally administered drug	190
considering the combined endotoxin exposure of the investigational drug and all	191
concomitantly administered drugs cited in the INDICATIONS AND USAGE section of	192
product labeling.	193
194
195

Contains Nonbinding Recommendations
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REFERENCES	196
197
Burrell R, Human Responses to Bacterial Endotoxin, 1994, Circ Shock, 43:137–153.	198
199
Hochstein HD, EA Fitzgerald, FG McMahon, and R Vargas, 1994, Properties of US Standard	200
Endotoxin (EC-5) in Human Male Volunteers. J Endotoxin Res, 1(1):52–56.	201
202
Reeve AJ, S Patel, A Fox, K Walker, and L Urban, 2000, Intrathecally Administered Endotoxin	203
or Cytokines Produce Allodynia, Hyperalgesia and Changes in Spinal Cord Neuronal Responses	204
to Nociceptive Stimuli in the Rat, Eur J Pain, 4(3):247–257.	205
206
Simmons RL, TB Ducker, and AM Martin Jr., 1969, Comparative Pathology after Intrathecal	207
Endotoxin in the Rabbit, Dog, and Monkey. Experentia, 25(6):622–623.	208
209
Suffredini AF, RE Fromm, MM Parker, M Brenner, JA Kovacs, RA Wesley, and JE Parrillo,	210
1989, The Cardiovascular Response of Normal Humans to the Administration of Endotoxin, N	211
Engl J Med, 321(5):280–287.	212
213
Suffredini AF and RJ Noveck, Human Endotoxin Administration as an Experimental Model in	214
Drug Development, 2014, Clin Pharmacol Ther, 96(4):418–422.	215