Guidance for Industry M4S: The CTD - Safety Appendices

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U.S. Department of Health and Human Services
Food and Drug Administration
Center for Drug Evaluation and Research (CDER)
Center for Biologics Evaluation and Research (CBER)
August 2001

ICH

Appendices

Additional copies are available from:

Office of Training and Communications
Division of Drug Information, HFD-240
Center for Drug Evaluation and Research
Food and Drug Administration
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Rockville, MD 20857
(Tel) 301-827-4573
(Internet) http://www.fda.gov/cder/guidance/index.htm

or

Office of Communication, Training and
Manufacturers Assistance, HFM-40
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Internet: http://www.fda.gov/cber/guidelines.htm.
Fax: 1-888-CBERFAX or 301-827-3844
Mail: the Voice Information System at 800-835-4709 or 301-827-1800

August 2001

ICH

APPENDIX A: EXAMPLES OF TABLES AND FIGURES FOR WRITTEN SUMMARIES

The tables and figures in Appendix A are presented merely as examples. Applicants should provide tables and figures using a format appropriate to the product.

Study references should be included in the table or text.

Tables should include statistics, if appropriate.

Table X: Binding of X and Its Major Metabolites and Comparators to Human X2 and X3 Receptors

 

Compound	X2 Ki1(nM)	X2 Ki2(nM)	X3 Ki1(nM)	X3 Ki2(nM)
1	538	2730	691	4550
2	2699	1050	2.0	181
3	578	14.4	141	10400
4	20	100	10.7	7.9
5	2100	3.1	281	28
6	7.5	8.4	44	2.8
7	3.11	3.76	1.94	1.93

K_i1 and K_i2 represent the high and low affinity binding sites, respectively (Data from Study Number).

Figure X: Blood Pressure Following Chronic Dosing With X to SHR^a



Blood pressure following chronic dosing with X to SHR^a[ref]. Hypotensive effect of saline i.v. infusion over 5 min (s) compared to X, 3 mg/kg i.v. infusion to SHR pretreated twice daily with saline, 1 mL/kg p.o., for 7 (m) or 14 (p) days or X, 25 mg/kg p.o., for 7 (l) or 14 (n) days. Saline pretreated statistical significances: p<0.05, all other points after challenge p<0.01. Values represent mean ± s.e.m.

^aSHR= spontaneous hypertensive rat (n=5 per group).

Table X: Model Independent Pharmacokinetic Parameters for X in Mice Following Single Oral Doses at 2, 10 and 30 mg/kg [ref]

 

Parameter (units)	Parameter value
Sex	Males			Females
Dose (mg/kg)	2	10	30	2	10	30
Cmax (ng/mL)	4.9	20.4	30.7	5.5	12.9	28.6
Tmax (h)	0.8	0.4	0.3	0.4	0.5	0.3
AUC0-t (ng.h/mL)	21.6	80.5	267	33.3	80	298
AUC0-inf (ng.h/mL)	28.3	112	297	40.2	90	327
Pharmacokinetic parameters were determined in pooled plasma from three animals at each time.

Table X: Excretion of Radioactive Material Following Single Doses of [14C]X to Male Mice [ref]

Dose (mg/kg)/	Percentage of administered dose
route	Urine*	Feces	Total+
2.8 i.v.	88.1 ± 7.4	5.5 ± 0.7	93.6 ± 6.9
8.8 p.o.	89.4 ± 4.7	6.9 ± 1.4	95.3 ± 3.4
Excretion was determined over 168 hours after dosing. Values are means ± S.D. (n= 5 for p.o. and 5 for i.v.) * - includes radioactivity in cage wash (22.1% after p.o. and 21.7% after i.v.). + - includes radioactivity in the carcass.

Table X: Concentrations of Radioactive Material in the Tissues of Male Rats After a Single Intravenous Dose of [14C]X at 1.75 mg/kg [refs]

 

Tissue	Concentration (ng equiv.*/g)
	1 h	6 h	24 h	48 h	72 h
Blood	105	96.6	2.34	2.34	3.65
Plasma	142	175	3.12	ND	ND
Adrenals	656	49.2	14.3	9.63	ND
Bone marrow	359	31.5	ND	ND	ND
Brain	116	9.37	ND	ND	ND
Eyes	124	28.9	4.69	ND	ND
Fat	490	44.0	10.2	6.25	5.47
Heart	105	26.6	ND	ND	ND
Kidneys	1280	651	21.6	13.3	9.63
Large intestine	570	2470	39.3	12.0	ND
Liver	875	380	133	87.7	64.6
Lungs	234	59.1	7.55	ND	ND
* - ng of X free base equivalent/g. N= 5 animals/time point. ND - Not detected.

Table X: Excretion of Radioactive Material Following Single Doses of [14C]X to Male Rats [refs]

 

Dose (mg/kg)/	Percentage of administered dose
route	Urine	Feces	Bile	Total
1.75 i.v.	61.3 ± 9.3	30.3 ± 4.1	-	95.2 ± 5.0
1.75 p.o.	57.4 ± 3.8	37.0 ± 3.4	-	95.2 ± 1.5
2 p.o.	72.3 ± 0.8	26.9 ± 1.9	-	99.5 ± 1.1
20 p.o.	23.5 ± 6.3	0.5 ± 0.2	76.0 ± 5.9	100 ± 0.8
220 p.o.	67.1 ± 9.0	24.8 ± 5.0	-	93.3 ± 6.8
Excretion was determined over 168 h period in Wistar rats:Values are means ± S.D. (n=5); - not assayed; Total includes radioactivity in the carcass and cage washings.

Table X: Comparative Pharmacokinetic Data and Systemic Exposure to X Following Oral Administration to Mice, Rats, Dogs, and Patients [ref]

Species (formulation)	Dose (mg/kg/day)	Systemic (plasma) exposure			References
		Cmax (ng/mL)	AUC (ng.h/mL)#
Man (tablet)	0.48$	36.7	557	X
Mouse (solution)	8.8	68.9 (1.9)*	72.7 (0.2)*	Y
	21.9	267 (7.3)*	207 (0.5)*
	43.8	430 (11.7)*	325 (0.7)*
Rat (solution)	50	479 (13.0)*	1580 (2.8)*	Z
Dogs (solution)	1.5	5.58 (0.2)*	15.9 (<0.1)*	V
	5	24.8 (0.7)*	69.3 (0.1)*
	15	184 (5.0)*	511 (0.9)*

Data presented are for male and female animals and are after daily repeated oral administration (at the end of the 60-day mouse study, 14-day rat study, and 1-year dog study). Data for man are extrapolated from dose normalized data obtained in male and female patients following t.i.d regimen.
# - AUC0-6 in the mouse, AUC0-t in the rat and in the dog and dose normalized AUC0-t x 24 in man.

$ - calculated from the total daily dose assuming a body weight of 50 kg for man.

* - Numbers in parentheses represent ratios of exposure in animals to those in patients.

Table X: Incidence of Proliferative Interstitial (Leydig) Cell Lesions in Rats [ref]

 

	Dose Groups
Lesion	Control	3 mg/kg	30 mg/kg	100 mg/kg
Hyperplasia (only)	x/50 (%)	x/50 (%)	x/50 (%)	x/50 (%)
Adenoma (only)	x/50 (%)	x/50 (%)	x/50 (%)	x/50 (%)
Adenoma + Hyperplasia	x/50 (%)	x/50 (%)	x/50(%)	x/50 (%)
Total*	x/50 (%)	x/50 (%)	x/50 (%)	x/50 (%)

* Adenoma and/or Hyperplasia.

APPENDIX B: THE NONCLINICAL TABULATED SUMMARIES TEMPLATES

2.6.3 Pharmacology

2.6.3.1 Pharmacology: Overview

2.6.3.2 Primary Pharmacodynamics*

2.6.3.3 Secondary Pharmacodynamics*

2.6.3.4 Safety Pharmacology

2.6.3.5 Pharmacodynamic Drug Interactions*

2.6.5 Pharmacokinetics

2.6.5.1 Pharmacokinetics: Overview

2.6.5.2 Analytical Methods and Validation Reports*

2.6.5.3 Pharmacokinetics: Absorption After a Single Dose

2.6.5.4 Pharmacokinetics: Absorption after Repeated Doses

2.6.5.5 Pharmacokinetics: Organ Distribution

2.6.5.6 Pharmacokinetics: Plasma Protein Binding

2.6.5.7 Pharmacokinetics: Study in Pregnant or Nursing Animals

2.6.5.8 Pharmacokinetics: Other Distribution Study

2.6.5.9 Pharmacokinetics: Metabolism In Vivo

2.6.5.10 Pharmacokinetics: Metabolism In Vitro

2.6.5.11 Pharmacokinetics: Possible Metabolic Pathways

2.6.5.12 Pharmacokinetics: Induction/Inhibition of Drug-Metabolizing Enzymes

2.6.5.13 Pharmacokinetics: Excretion

2.6.5.14 Pharmacokinetics: Excretion into Bile

2.6.5.15 Pharmacokinetics: Drug-Drug Interactions

2.6.5.16 Pharmacokinetics: Other

2.6.7 Toxicology

2.6.7.1 Toxicology: Overview

2.6.7.2 Toxicokinetics: Overview of Toxicokinetics Studies

2.6.7.3 Toxicokinetics: Overview of Toxicokinetics Data

2.6.7.4 Toxicology: Drug Substance

2.6.7.5 Single-Dose Toxicity

2.6.7.6 Repeat-Dose Toxicity: Nonpivotal Studies

2.6.7.7 Repeat-Dose Toxicity: Pivotal Studies

2.6.7.8 Genotoxicity: In Vitro

2.6.7.9 Genotoxicity: In Vivo

2.6.7.10 Carcinogenicity

2.6.7.11 Reproductive and Developmental Toxicity: Nonpivotal Studies

2.6.7.12Reproductive and Developmental Toxicity: Fertility and Early Embryonic Development to Implantation (Pivotal)

2.6.7.13Reproductive and Developmental Toxicity: Effects on Embryofetal Development (Pivotal)

2.6.7.14Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotol)

2.6.7.15 Studies in Juvenile Animals^a (template not provided; see footnote a)

2.6.7.16 Local Tolerance

2.6.7.17 Other Toxicity Studies

* : Tabulated summary is optional. It is preferable to include text tables and figures with the Nonclinical Written Summary.

^a : When a juvenile animal study has been conducted, it should be tabulated using the template appropriate for the type of study and located in Section 2.6.7.15.

2.6.3.1 Pharmacology Overview Test Article: (1)

Type of Study	Test System	Method of Administration	Testing Facility	Study Number(4)	Location Vol. Page
Primary Pharmacodynamics (2)					(3)
Secondary Pharmacodynamics
Safety Pharmacology
Pharmacodynamic Drug Interactions

Notes: (1) International Nonproprietary Name (INN)

(2) There should be one line for each pharmacology report, in the same order as the CTD. Reports that contain a GLP Compliance Statement should be identified in a footnote.

(3) The location of the Technical Report in the CTD should be indicated.

(4) Or Report Number (on all tables).

2.6.3.4 Safety Pharmacology(1) Test Article: (2)

Organ Systems Evaluated	Species/ Strain	Method of Admin.	Dosesa (mg/kg)	Gender and No. per Group	Noteworthy Findings	GLP Compliance	Study Number(3)

Notes: (1) All safety pharmacology studies should be summarized.

(2) International Nonproprietary Name (INN).

(3) Or Report Number (on all tables).

a - Single dose unless specified otherwise.

2.6.5.1 Pharmacokinetics Overview Test Article: (1)

Type of Study	Test System	Method of Administration	Testing Facility	Study Number	Location Vol. Page
Absorption (2)					(3)
Distribution
Metabolism
Excretion
Pharmacokinetic Drug Interactions
Other

Notes: (1) International Nonproprietary Name (INN).

(2) There should be one line for each pharmacokinetics report, in the same order as the CTD. Reports that contain a GLP Compliance Statement should be identified in a footnote.

(3) The location of the Technical Report in the CTD should be indicated.

2.6.5.3 Pharmacokinetics: Absorption After a Single Dose Test Article: (1)

Location in CTD: Vol. Page

Study No.

Species	__________	__________	__________	__________	_________
Gender (M/F)/Number of animals	(4)
Feeding condition
Vehicle/Formulation
Method of Administration
Dose (mg/kg)
Sample (e.g., whole blood, plasma, serum)
Analyte
Assay (2)
PK parameters:
Additional Information: (3) Notes: (1) International Nonproprietary Name (INN). (2) For example, HPLC, LSC with 14C-labeled compound. (3) For example, brief textual results, species differences, gender differences, dose dependency, or special comments. (4) There should be one column for each study conducted. For comparison, representative information on humans at the maximum recommended dose should be included.

2.6.5.4 Pharmacokinetics: Absorption after Repeated Doses Test Article:

[Data can be tabulated as in the format of 2.6.5.3 if applicable.]

Format A

2.6.5.5 Pharmacokinetics: Organ Distribution Test Article:

Location in CTD: Vol. Page

Study No.

Species:
Gender (M/F)/Number of animals:
Feeding condition:
Vehicle/Formulation:
Method of Administration:
Dose (mg/kg):
Radionuclide:
Specific Activity:
Sampling time:
	Concentration (unit)
Tissues/organs	T(1)	T(2)	T(3)	T(4)	T(5)	t1/2?
Additional information:

Alternate Format B

2.6.5.5 Pharmacokinetics: Organ Distribution Test Article:

Location in CTD: Vol. Page

Study No.

Species:
Gender (M/F)/Number of animals:
Feeding condition:
Vehicle/Formulation:
Method of Administration:
Dose (mg/kg):
Radionuclide:
Specific Activity:
Analyte/Assay (unit):
Sampling time:
	Ct		Last time point
Tissues/organs	conc.	T/P1)	conc.	T/P1)	Time	AUC	t1/2?
Additional information:

¹⁾ [Tissue]/[Plasma]

2.6.5.6 Pharmacokinetics: Plasma Protein Binding Test Article:

Study system:
Target entity, Test system and method:
Species	Conc. tested	% Bound		Study No.	Location in CTD Vol. Page
Additional Information:

2.6.5.7 Pharmacokinetics: Study in Pregnant or Nursing Animals (1) Test Article: (2)

Location in CTD: Vol. Page

Placental transfer Study No.

Species:
Gestation day/Number of animals:
Vehicle/Formulation:
Method of Administration:
Dose (mg/kg):
Analyte:
Assay:
Time (hr)		__________	_________	_________	__________	__________
Concentration/Amount (% of dose) Dam (3): Fetus (3):
Additional Information:

Location in CTD: Vol. Page

Excretion into milk Study No.

Species:
Lactating date/Number of animals:
Feeding condition:
Vehicle/Formulation:
Method of Administration:
Dose (mg/kg):
Analyte:
Assay:
Time [hr]	__________	__________	__________	__________	__________
Concentration: Milk: Plasma: Milk/plasma: Neonates:
Additional Information:

Notes for Table 2.6.5.7

(1) Even if the data are obtained in reproduction toxicology studies, they should be presented in this table.

(2) International Nonproprietary Name (INN).

(3) The tissue sampled should be described (e.g., plasma for dams, fetal concentrations).

2.6.5.8 Pharmacokinetics: Other Distribution Study Test Article:

2.6.5.9 Pharmacokinetics: Metabolism In Vivo Test Article:

Gender(M/F)/Number of animals:
Feeding condition:
Vehicle/Formulation:
Method of Administration:
Dose (mg/kg):
Radionuclide:
Specific Activity:
				% of Compound in Sample					Location in CTD
Species	Sample	Sampling Time or Period	% of Dose in Sample	Parent_		M1	M2	Study No.	Vol	Page
	Plasma Urine Bile Feces
	Plasma Urine Bile Feces
	Plasma Urine Bile Feces
Additional Information: Note: Human data should be included for comparison if available.

2.6.5.10 Pharmacokinetics: Metabolism In Vitro Test Article:

Location in CTD: Vol. Page

Study No.

Study system:
Time	__________	__________	__________	__________	__________
Concentration:
Compounds Parent M-1 M-2
Additional Information: Note: Human data should be included for comparison if available.

2.6.5.11 Pharmacokinetics: Possible Metabolic Pathways Test Article:

(Illustrate possible metabolic map indicating species in which metabolic reactions occur.)

2.6.5.12 Pharmacokinetics: Induction/Inhibition of Drug-Metabolizing Enzymes Test Article:

Location in CTD: Vol. Page

Study No.

Note: Nonclinical studies only.

Type of study:

Method:

Tabulated results:

Additional Information:

2.6.5.13 Pharmacokinetics: Excretion Test Article: (1)

Species

__________

__________

__________

_________

Gender (M/F)/Number of animals

(3)

Feeding condition

Vehicle/Formulation

Method of Administration

Dose (mg/kg)

Analyte

Assay

Excretion route (4)

Urine

Feces

Total

Urine

Feces

Total

Urine

Feces

Total

Urine

Feces

Total

Time 0 - T hr

Study number

Location in CTD

Additional Information: (2) Notes: (1) International Nonproprietary Name (INN). (2)For example, brief textual results, species differences, gender differences, dose dependency, or special comments. (3) There should be one column for each study conducted. For comparison, representative information on humans at the maximum recommended dose should be included. Can be combined with the Absorption Table if appropriate. (4) Other routes (e.g., biliary, respiratory) should be added, if performed.

2.6.5.14 Pharmacokinetics: Excretion into Bile Test Article:

[Data can be tabulated as in the format of 2.6.5.13 if applicable.]

2.6.5.15 Pharmacokinetics: Drug-Drug Interactions Test Article:

Location in CTD: Vol. Page

Study No.

Type of study:

Method:

Tabulated results:

Additional Information:

2.6.5.16 Pharmacokinetics: Other Test Article:

Location in CTD: Vol. Page

Study No.

Type of study:

Method:

Tabulated results:

Additional Information:

2.6.7.1 Toxicology Overview Test Article: (1)

Type of Study	Species and Strain	Method of Administration	Duration of Dosing	Doses (mg/kga)	GLP Compliance	Testing Facility	Study Number	Location Vol. Page
Single-Dose Toxicity	(2)							(3)
Repeat-Dose Toxicity
Genotoxicity
Carcinogenicity
Reproductive and Developmental Toxicity
Local Tolerance
Other Toxicity Studies

Notes: (1) International Nonproprietary Name (INN).

(2) There should be one line for each toxicology report, in the same order as the CTD.

(3) The location of the Technical Report in the CTD should be indicated.

a - Unless otherwise specified. For Repeat-Dose Toxicity, the highest No Observed Adverse Effect Level (NOAEL) is underlined.

2.6.7.2 Toxicokinetics Overview of Toxicokinetics Studies Test Article: (1)

Type of Study	Test System	Method of Administration	Doses (mg/kg)	GLP Compliance	Study Number	Location Vol. Page
(2)						(3)

Notes: (1) International Nonproprietary Name (INN).

(2) There should be one line for each toxicokinetics report, in the same order as the CTD (Section 3, Toxicology).

(3) The location of the Technical Report in the CTD should be indicated.

2.6.7.3 Toxicokinetics Overview of Toxicokinetics Data Test Article: (1)

(2)

Notes: (1) International Nonproprietary Name (INN).

(2) A one- to three-page summary (tables and/or figures) of steady state toxicokinetic data should be prepared in a format that facilitates comparisons across species, including humans.

2.6.7.4 Toxicology Drug Substance Test Article: (1)

Batch No.	Purity (%)	Specified Impurities ( )			Study Number	Type of Study
PROPOSED SPECIFICATION:
(2)						(3)

Notes: (1) International Nonproprietary Name (INN).

(2) All batches used in the Toxicology studies should be listed in approximate chronological order.

(3) The Toxicology studies in which each batch was used should be identified.

2.6.7.5 Single-Dose Toxicity (1) Test Article: (2)

Species/ Strain	Method of Administration (Vehicle/ Formulation)	Doses (mg/kg)	Gender and No. per Group	Observed Maximum Nonlethal Dose (mg/kg)	Approximate Lethal Dose (mg/kg)	Noteworthy Findings	Study Number

Notes: (1) All single-dose toxicity studies should be summarized, in the same order as the CTD. Footnotes should be used to indicate special features, such as unusual duration, infusion rate, or age of test subjects.

(2) International Nonproprietary Name (INN).

2.6.7.6 Repeat-Dose Toxicity Nonpivotal Studies (1) Test Article: (2)

Species/ Strain	Method of Administration (Vehicle/ Formulation)	Duration of Dosing	Doses (mg/kg)	Gender and No. per Group	NOAELa (mg/kg)	Noteworthy Findings	Study Number

Notes: (1) All repeat-dose toxicity studies (including all range-finding toxicity studies), other than the definitive GLP studies specified by ICH Guidance M3 Nonclinical Safety Studies for the Conduct of Human Clinical Trials for Pharmacaeuticals (November 1997), should be summarized in the same order as the CTD. Footnotes should be used to indicate special features, such as unusual age of test subjects.

(2) International Nonproprietary Name (INN).

________

a - No Observed Adverse Effect Level.

2.6.7.7 (1) Repeat-Dose Toxicity (2) Report Title: Test Article: (3)

Species/Strain: Duration of Dosing: Study No.

Initial Age: Duration of Postdose: Location in CTD: Vol. Page

Date of First Dose: Method of Administration:

Vehicle/Formulation: GLP Compliance:

Special Features:

No Observed Adverse Effect Level:

Daily Dose (mg/kg)	0 (Control)
Number of Animals Toxicokinetics: AUC ( ) (4) Noteworthy Findings Died or Sacrificed Moribund Body Weight (%a) Food Consumption (%a) Water Consumption ( ) Clinical Observations Ophthalmoscopy Electrocardiography	M: (5) (5) (5)	F:	M:	F:	M:	F:	M:	F:

- No noteworthy findings. + Mild ++ Moderate +++ Marked (6)

(7) * - p<0.05 ** - p<0.01

a - At end of dosing period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences).

(Continued)

2.6.7.7 (1) Repeat-Dose Toxicity Study No. (Continued)

Daily Dose (mg/kg)	0 (Control)
Number of Animals	M:	F:	M:	F:	M:	F:	M:	F:
Hematology
Serum Chemistry
Urinalysis
Organ Weightsa (%)
Gross Pathology
Histopathology
Additional Examinations
Postdose Evaluation: Number Evaluated (8) (9)

- No noteworthy findings.

(7) * - p<0.05 ** - p<0.01

a - Both absolute and relative weights differed from controls in the direction indicated. Number indicates percent difference for the absolute organ weights.

Notes for Table 2.6.7.7

(1) The tables should be numbered consecutively (e.g., 2.6.7.7A, 2.6.7.7B, 2.6.7.7C).

(2) There should be one table for each of the repeat-dose toxicity studies specified by ICH Guidance M3 Nonclinical Safety Studies for the Conduct of Human Clinical Trials for Pharmacaeuticals (November 1997), as well as any other repeat-dose toxicity studies that could be considered pivotal.

(3) International Nonproprietary Name (INN).

(4) Steady state AUC, Cmax, Css, or other toxicokinetic information supporting the study. If from a separate study, the study number should be given in a footnote.

(5) ONLY NOTEWORTHY FINDINGS SHOULD BE PRESENTED. If additional parameters (other than those in the template) showed noteworthy changes, these should be added to the tables. In general, data at end of dosing period can be shown; however, if there were additional noteworthy findings at earlier timepoints, these should be included. Footnotes should be used as needed to provide additional information about the tests or the results.

(6) Or other scale, as appropriate.

(7) Methods of statistical analyses should be indicated.

(8) All parameters that still show drug-related changes should be listed. This section should be deleted if the study does not include a postdose evaluation.

(9) When appropriate, information on animals that were necropsied early should be presented separately.

2.6.7.8 (1) Genotoxicity: In Vitro Report Title: Test Article: (2)

Test for Induction of: No. of Independent Assays: Study No.

Strains: No. of Replicate Cultures: Location in CTD: Vol. Page

Metabolizing System: No. of Cells Analyzed/Culture:

Vehicles: For Test Article: For Positive Controls: GLP Compliance:

Treatment: Date of Treatment:

Cytotoxic Effects:

Genotoxic Effects:

Metabolic Activation	Test Article	Concentration or Dose Level ( (3) )	_____________	_____________	_____________	____________	_____________
Without Activation		(4)
With Activation

Notes: (1) The tables should be numbered consecutively (e.g.,2.6.7.8A, 2.6.7.8B). Results of replicate assays should be shown on subsequent pages.

(2) International Nonproprietary Name (INN).

(3) Units should be inserted.

(4) If precipitation is observed, this should be indicated in a footnote.

(5) Methods of statistical analyses should be indicated.

(5) * - p<0.05 ** - p<0.01

2.6.7.9 (1) Genotoxicity: In Vivo Report Title: Test Article: (2)

Test for Induction of: Treatment Schedule: Study No.

Species/Strain: Sampling Time: Location in CTD: Vol. Page

Age: Method of Administration:

Cells Evaluated: Vehicle/Formulation: GLP Compliance:

No. of Cells Analyzed/Animal: Date of Dosing:

Special Features:

Toxic/Cytotoxic Effects:

Genotoxic Effects:

Evidence of Exposure:

Test Article	Dose (mg/kg)	No. of Animals	______________	______________	______________	_____________

Notes: (1) The tables should be numbered consecutively (e.g.,2.6.7.9A, 2.6.7.9B).

(2) International Nonproprietary Name (INN).

(3) Methods of statistical analysis should be indicated.

(3) * - p<0.05 ** - p<0.01).

2.6.7.10 (1) Carcinogenicity Report Title: Test Article: (2)

Species/Strain: Duration of Dosing: Study No.

Initial Age: Method of Administration: Location in CTD: Vol. Page

Date of First Dose: Vehicle/Formulation:

Treatment of Controls: GLP Compliance:

Basis for High-Dose Selection: (3)

Special Features:

Daily Dose (mg/kg)	0 (Control)
Gender Toxicokinetics: AUC ( ) (4) Number of Animals At Start Died/Sacrificed Moribund Terminal Sacrifice Survival (%) Body Weight (%a) Food Consumption (%a)	M (5)	F	M	F	M	F	M	F

(6) * - p<0.05 ** - p<0.01

a - At 6 months. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences). (Continued)

2.6.7.10 (1) Carcinogenicity Study No. (Continued)

Daily Dose (mg/kg)	(Control)		0 (Control)
Number Evaluated Number of Animals with Neoplastic Lesions: (7) Noteworthy Findings: Gross Pathology Histopathology - Non-Neoplastic Lesions	M:	F:	M:	F:	M:	F:	M:	F:	M:	F:

- No noteworthy findings.

* - p<0.05 ** - p<0.01

Notes for Table 2.6.7.10

(1) Tables should be numbered consecutively (e.g., 2.6.7.10A, 2.6.7.10B). There should be one table for each carcinogenicity study.

(2) International Nonproprietary Name (INN).

(3) From ICH Guidance S1C Dose Selection for Carcinogenicity Studies of Pharmaceuticals (March 1995).

(4) Steady state AUC, Cmax, Css, or other toxicokinetic information supporting the study. If the information is from a separate study, the Study Number should be given in a footnote.

(5) If additional parameters showed drug-related changes, these should be added to the tables. Footnotes should be used as needed to provide additional information about the tests or the results.

(6) Methods of statistical analysis should be indicated.

(7) Drug-related lesions should be listed first. Then other lesions should be listed by alphabetically ordered organs and/or tissues.

2.6.7.11 Reproductive and Developmental Toxicity Nonpivotal Studies (1) Test Article: (2)

Species/ Strain

Method of Administration (Vehicle/ Formulation)

Dosing Period

Doses mg/kg

No. per Group

Noteworthy Findings

Study Number

Notes: (1) All reproduction toxicity studies (including all relevant range-finding studies), other than the definitive GLP studies specified by M3 Nonclinical Safety Studies for the Conduct of Human Clinical Trials for Pharmaceuticals, November 1997, should be summarized in the same order as the CTD. However, investigative studies should be summarized using a more detailed template.

(2) International Nonproprietary Name (INN).

2.6.7.12 (1) Reproductive and Developmental Toxicity - Report Title: Test Article: (2)

Fertility and Early Embryonic

Development to Implantation (3)

Design similar to ICH 4.1.1? Duration of Dosing:M: Study No.

Species/Strain: Day of Mating: (8)F: Location in CTD: Vol. Page

Initial Age: Day of C-Section:

Date of First Dose: Method of Administration: GLP Compliance:

Special Features: Vehicle/Formulation:

No Observed Adverse Effect Level:

F₀ Males:

F₀ Females:

F₁ Litters:

Daily Dose (mg/kg)	0 (Control)
Males Toxicokinetics: AUC ( ) (4) No. Evaluated No. Died or Sacrificed Moribund Clinical Observations Necropsy Observations Body Weight (%a) Food Consumption (%a) Mean No. Days Prior to Mating No. of Males that Mated No. of Fertile Males	(5)

-No noteworthy findings. + Mild ++Moderate +++Marked (6)

(7) *- p<0.05 ** - p<0.01

a - After 4 weeks of dosing. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences). (Continued)

2.6.7.12 (1) Reproductive and Developmental Toxicity Study No. (Continued)

 

Daily Dose (mg/kg)	0 (Control)
Females Toxicokinetics: AUC ( ) (4)   No. Evaluated No. Died or Sacrificed Moribund Clinical Observations Necropsy Observations Premating Body Weight (%a) Gestation Body Weight (%a) Premating Food Consumption (%a) Gestation Food Consumption (%a) Mean No. Estrous Cycles/14 days Mean No. Days Prior to Mating No. of Females Sperm Positive No. of Pregnant Females No. Aborted or with Total Resorption of Litter Mean No. Corpora Lutea Mean No. Implantations Mean % Preimplantation Loss Mean No. Live Conceptuses Mean No. Resorptions No. Dead Conceptuses Mean % Postimplantation Loss

-No noteworthy findings. + Mild ++Moderate +++Marked (6)

(7)* - p<0.05 ** - p<0.01

a - At end of premating or gestation period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences).

Notes for Tables 2.6.7.12, 2.6.7.13, and 2.6.7.14

(1) If there are multiple studies of this type, the tables should be numbered consecutively (e.g., 2.6.7.12A, 2.6.7.12B, 2.6.7.13A, 2.6.7.13B).

(2) International Nonproprietary Name (INN).

(3) If a modified study design is used, tables should be modified accordingly.

(4) Steady state AUC, Cmax, or other toxicokinetic information supporting the study. If the information is from a separate study, the study number should be given in a footnote.

(5) POSSIBLE PRESENTATIONS OF THE RESULTS ARE SHOWN IN THESE TEMPLATES. DATA PRESENTATION SHOULD BE FLEXIBLE AND APPROPRIATE ACCORDING TO OPTIMAL STATISTICAL ANALYSIS AND THE DESIGN OF THE STUDY. If additional parameters showed drug-related changes, these should be added to the tables. Footnotes should be used as needed to provide additional information about the tests or the results.

(6) Or other scale as appropriate.

(7) Methods of statistical analysis should be indicated.

(8) Day of mating should be indicated (e.g., Day 0 or Day 1).

2.6.7.13 (1) Reproductive and Developmental Toxicity - Report Title: Test Article: (2)

Effects on Embryofetal

Development (3)

Design similar to ICH 4.1.3? Duration of Dosing: Study No.

Day of Mating: (8)

Species/Strain: Day of C-Section: Location in CTD: Vol. Page

Initial Age: Method of Administration:

Date of First Dose: Vehicle/Formulation: GLP Compliance:

Special Features:

No Observed Adverse Effect Level:

F₀ Females:

F₁ Litters:

Daily Dose (mg/kg)	0 (Control)
Dams/Does: Toxicokinetics: AUC () (4) No. Pregnant No. Died or Sacrificed Moribund No. Aborted or with Total Resorption of Litter Clinical Observations Necropsy Observations Body Weight (%a) Food Consumption (%a) Mean No. Corpora Lutea Mean No. Implantations Mean % Preimplantation Loss	(5)

- No noteworthy findings. + Mild ++Moderate +++Marked (6) G = Gestation day

(7) * - p<0.05 ** - p<0.01

a- At end of dosing period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences). (Continued)

2.6.7.13 (1) Reproductive and Developmental Toxicity Study No. (Continued)

Daily Dose (mg/kg)	0 (Control)
Litters: No. Litters Evaluated No. Live Fetuses Mean No. Resorptions No. of Litters with Dead Fetuses Mean % Postimplantation Loss Mean Fetal Body Weight (g) Fetal Sex Ratios Fetal Anomalies: Gross External Visceral Anomalies Skeletal Anomalies Total Affected Fetuses (Litters)

- No noteworthy findings.

* - p<0.05 ** - p<0.01

2.6.7.14 (1) Reproductive and Developmental Toxicity - Report Title: Test Article: (2)

Effects on Pre- and Postnatal

Development, Including Maternal Function (3)

Design similar to ICH 4.1.2? Duration of Dosing: Study No.

Day of Mating: (8)

Species/Strain: Method of Administration: Location in CTD: Vol. Page

Initial Age Vehicle/Formulation:

Date of First Dose: Litters Culled/Not Culled: GLP Compliance:

Special Features:

No Observed Adverse Effect Level:

F₀ Females:

F₁ Males:

F₁ Females:

Daily Dose (mg/kg)	0 (Control)
F0 Females: Toxicokinetics: AUC ( ) (4) No. Pregnant No. Died or Sacrificed Moribund No. Aborted or with Total Res. of Litter Clinical Observations Necropsy Observations Gestation Body Weight (%a) Lactation Body Weight (%a) Gestation Food Consumption (%a) Lactation Food Consumption (%a) Mean Duration of Gestation (days) Abnormal Parturition	(5)

- No noteworthy findings. + Mild ++Moderate +++Marked (6) G = Gestation day L = Lactation day

(7) * - p<0.05 ** - p<0.01)

a - At end of gestation or lactation. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences). (Continued)

2.6.7.14 (1) Reproductive and Developmental Toxicity Study No. (Continued)

Daily Dose (mg/kg)		0 (Control)
F1 Litters: (Preweaning) F1 Males: (Postweaning)	No. Litters Evaluated Mean No. of Implantations Mean No. Pups/Litter Mean No. Liveborn Pups/Litter No. of Litters with Stillborn Pups Postnatal Survival to Day 4 Postnatal Survival to Weaning No. of Total Litter Losses Change in Pup Body Weightsa (g) Pup Sex Ratios Pup Clinical Signs Pup Necropsy Observations No. Evaluated Postweaning Per Litter No. Died or Sacrificed Moribund Clinical Observations Necropsy Observations Body Weight Changeb (g) Food Consumption (%c) Preputial Separation Sensory Function Motor Activity Learning and Memory Mean No. Days Prior to Mating No. of Males that Mated No. of Fertile Males

- No noteworthy findings. + Mild ++Moderate +++Marked (6)

(7)* - p<0.05 ** - p<0.01

a - From birth to weaning.

b - From weaning to mating.

c - At end of postweaning period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences).

2.6.7.14 (1) Reproductive and Developmental Toxicity Study No. (Continued)

Daily Dose (mg/kg)		0 (Control)
F1 Females: (Postweaning) F2 Litters:	No. Evaluated Postweaning No. Died or Sacrificed Moribund Clinical Observations Necropsy Observations Premating Body Weight Changea (g) Gestation Body Weight Change (g) Premating Food Consumption (%b) Gestation Food Consumption (%b) Mean Age of Vaginal Patency (days) Sensory Function Motor Activity Learning and Memory Mean No. Days Prior to Mating No. of Females Sperm-Positive No. of Pregnant Females Mean No. Corpora Lutea Mean No. Implantations Mean % Preimplantation Loss Mean No. Live Conceptuses/Litter Mean No. Resorptions No. of Litter with Dead Conceptuses No. Dead Conceptuses Mean % Postimplantation Loss Fetal Body Weights (g) Fetal Sex Ratios (% males) Fetal Anomalies

- No noteworthy findings. + Mild ++Moderate +++Marked (6)

(7)* - p<0.05 ** - p<0.01

a - From weaning to mating

b - At end of premating or gestation period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences).

2.6.7.14 (1) Reproductive and Developmental Toxicity Study No. (Continued)

Daily Dose (mg/kg)		0 (Control)
F1 Females: (Postweaning) F2 Litters:	No. Evaluated Postweaning No. Died or Sacrificed Moribund Clinical Observations Necropsy Observations Premating Body Weight Changea (g) Gestation Body Weight Change (g) Premating Food Consumption (%b) Gestation Food Consumption (%ab) Mean Age of Vaginal Patency (days) Sensory Function Motor Activity Learning and Memory Mean No. Days Prior to Mating No. of Females Sperm Positive No. of Pregnant Females Mean Duration of Gestation Abnormal Parturition No. Litters Evaluated Mean No. of Implantations Mean No. Pups/Litter Mean No. Liveborn Pups/Litter Mean No. Stillborn Pups/Litter Postnatal Survival to Day 4 Postnatal Survival to Weaning Change in Pup Body Weightsa (g) Pup Sex Ratios Pup Clinical Signs Pup Necropsy Observations	Note: Alternate Format for Natural Parturition.

- No noteworthy findings. + Mild ++Moderate +++Marked (6)

(7)* - p<0.05 ** - p<0.01

a - From birth to mating.

b - At end of premating or gestation period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is based on actual data (not on the percent differences).

2.6.7.16 Local Tolerance (1) Test Article: (2)

Species/ Strain	Method of Administration	Doses (mg/kg)	Gender and No. per Group	Noteworthy Findings	Study Number

Notes: (1) All local tolerance studies should be summarized.

(2) International Nonproprietary Name (INN).

2.6.7.17 Other Toxicity Studies (1) Test Article: (2)

Species/ Strain

Method of Administration

Duration of Dosing

Doses (mg/kg)

Gender and No. per Group

Noteworthy Findings

Study Number

Notes: (1) All supplementary toxicity studies should be summarized.

(2) International Nonproprietary Name (INN)

APPENDIX C: THE NONCLINICAL TABULALTED SUMMARIES - EXAMPLES

(The following examples correspond to the templates in Appendix B; examples are not provided for the templates Studies in Juvenile Animals or Local Tolerance)

EXAMPLE

2.6.3.1 Pharmacology Overview Test Article: Curitol Sodium

Type of Study	Test System	Method of Administration	Testing Facility	Study Number	Location Vol. Page
Primary Pharmacodynamics Antiviral activity vs. VZV Antiviral activity vs. VZV Antiviral activity vs. HSV Antiviral activity vs. CMV Antiviral activity vs. VZV Antiviral activity vs. SVV	Human embryonic lung fibroblasts Clinical isolates Human embryonic lung fibroblasts Human embryonic lung fibroblasts ICR mice African Green monkeys	In vitro In vitro In vitro In vitro Gavage Nasogastric Intubation	Sponsor Inc. Sponsor Inc. Sponsor Inc. Sponsor Inc. Sponsor Inc. Sponsor Inc.	95401 95402 95406 95408 95411 95420	1 1 1 1 1 1	1 20 30 45 55 100
Secondary Pharmacodynamics Antimicrobial activity	Gram positive and gram negative bacteria; yeasts	In vitro	Sponsor Inc.	95602	1	200
Safety Pharmacology Effects on central nervous systema Effects on cardiovascular system	Mice, rats, rabbits, and cats Dogs	Gavage Gavage, i.v.	Sponsor Inc. Sponsor Inc.	95703 95706	2 2	1 75
Pharmacodynamic Drug Interactions Interactions with anti-HIV activity of AZT	Human T lymphocytes	In vitro	Sponsor Inc.	95425	2	200

a - Report contains a GLP Compliance Statement.

EXAMPLE

2.6.3.4 Safety Pharmacology Test Article: Curitol Sodium

Organ Systems Evaluated	Species/ Strain	Method of Admin.	Dosesa (mg/kg)	Gender and No. per Group	Noteworthy Findings	GLP Compliance	Study Number
CNS	CD-1 Mice	Gavage	0, 10, 50, 250	10M	Slight prolongation of hexobarbital anesthesia (_10 mg/kg). No analgesic, anticonvulsive, or cataleptic properties. No effects on coordination, traction, or spontaneous motility.	Yes	92201
Renal, GI, CNS, and Hemostasis	CD-1 Mice	Gavage	0, 10, 50, 250	6M	Slight increases in urinary excretion of sodium and potassium (_50 mg/kg). No effects on GI transit time (charcoal meal), pupillary diameter, blood coagulation time, or urine volume.	No	92205
Cardiovascular	Mongrel Dogs	Intravenous	0, 3, 10, 30	3M	Dose-related transient decreases in blood pressure and increases in heart rate and respiratory rate (all doses). Minor ECG changes at 30 mg/kg. No effects on cardiac output, stroke volume, or total peripheral resistance.	Yes	92210

a - Single dose unless specified otherwise.

EXAMPLE

2.6.5.1 Pharmacokinetics Overview Test Article: Curitol Sodium

Type of Study	Test System	Method of Administration	Testing Facility	Study Number	Location Vol. Page
Absorption Absorption and excretion Absorption and excretion Absorption and excretion	Rats Dogs Monkeys	Gavage, i.v. Gavage, i.v. Gavage, i.v.	Sponsor Inc. Sponsor Inc. Sponsor Inc.	93302 93304 93306	1 1 1	1 25 50
Distribution Single-dose tissue distribution Repeat-dose tissue distribution Plasma protein binding Plasma protein binding	Rats Rats Mice, rats, dogs, monkeys, Humans, rats, dogs	Gavage Gavage In vitro Tablets/Gavage/ Capsules	Sponsor Inc. Sponsor Inc. Sponsor Inc. Sponsor Inc.	93307 93308 93311 93312	1 1 1 1	100 125 150 200
Metabolism Metabolites in blood, urine, and feces Metabolites in blood, urine, and feces	Rats Dogs	Gavage Gavage	Sponsor Inc. Sponsor Inc.	93402 93407	1 1	250 300
Excretion Absorption and excretion Absorption and excretion Absorption and excretion	Rats Dogs Monkeys	Gavage, i.v. Gavage, i.v. Gavage, i.v.	Sponsor Inc. Sponsor Inc. Sponsor Inc.	93302 93304 93306	1 1 1	1 25 50
Pharmacokinetic Drug Interactions Interaction with AZTa	Rats	Gavage	Sponsor Inc.	94051	1	350

a - Report contains a GLP Compliance Statement.

EXAMPLE

2.6.5.3 Pharmacokinetics: Absorption After a Single Dose Test Article: Curitol Sodium

Location in CTD Volume 1, Page 258

Study number 95104

Species	Mouse	Rat	Dog	Monkey	Human
Gender (M/F)/Number of animals	4M	3M	4F	2M	6M
Feeding condition	Fed	Fasted	Fasted	Fed	Fasted
Vehicle/Formulation	Suspension 10% acacia	Suspension 10% acacia	Capsule	Suspension 10% acacia	Tablet
Method of Administration	Gavage	Gavage	Capsule	Gavage	Oral
Dose (mg/kg)	15	8	5	5	4 mg
Sample (e.g., whole blood, plasma, serum)	Plasma	Plasma	Plasma	Plasma	Plasma
Analyte	TRAa	MM-180801	MM-180801	MM-180801	MM-180801
Assay	LSC	HPLC	HPLC	HPLC	HPLC
PK parameters:
Tmax (hr)	4.0	1.0	3.3	1.0	6.8
Cmax (ng/ml or ng-eq/ml)	2,260	609	172	72	8.2
AUC (ng or ng-eq x hr/ml)	15,201	2,579	1,923	582	135
(Time for calculation - hr)	(0-72)	(0-24)	(0.5-48)	(0-12)	(0-24)
T 1/2 (hr)	10.6	3.3	9.2	3.2	30.9
(Time for calculation - hr)	(7-48)	(1-24)	(24-96)	(1-12)	(24-120)
Additional Information: A single oral dose was well absorbed in mice, rats, dogs, and monkeys. In a study examining the concentration of compound in the portal vein and inferior vena cava, 30 minutes after a dose to rats, the concentration of compound was approximately 15-fold higher in the portal circulation compared to systemic circulation. This result indicated extensive metabolism and/or biliary secretion of compound in the rat. a - Total radioactivity, 14C

EXAMPLE

Format A

2.6.5.5 Pharmacokinetics: Organ Distribution Test Article: Curitol Sodium

Location in CTD: Vol.21 Page 1

Study No. 95207

Species: Rat
Gender (M/F)/Number of animals: 3M/each time point
Feeding condition: Fasted
Vehicle/Formulation: Solution/Water
Method of Administration: Oral Gavage
Dose (mg/kg): 10
Radionuclide: 14C
Specific Activity: 2x105 Bq/mg
Sampling time: 0.25, 0.5, 2, 6, 24, 96, and 192 hr
	Concentration (mcg/mL)
Tissues/organs	0.25	0.5	2	6	24	t1/2
Blood	9.2	3.7	1.8	0.9	0.1
Plasma	16.5	7.1	3.2	1.6	0.2
Brain	0.3	0.3	0.2	0.1	nd
Lung	9.6	14.1	7.3	2.9	0.1
Liver	73.0	54.5	19.9	12.4	3.2
Kidney	9.6	13.2	4.9	3.8	0.6
Testis	0.3	0.5	0.6	0.5	0.1
Muscle	1.0	1.2	0.8	0.3	nd
Additional information: Tissues and organs such as the heart, thymus, adrenal, spleen, stomach, intestine.....are examined but not shown. nd = Not detected.

EXAMPLE

Alternate Format B

2.6.5.5 Pharmacokinetics: Organ Distribution Test Article: Curitol Sodium

Location in CTD: Vol. 21 Page 1

Study No. 95207

Species: Rat
Gender (M/F) / Number of animals: 3M/each time point
Feeding condition: Fed
Vehicle/Formulation: Solution/Saline
Method of Administration: Intravenous
Dose (mg/kg): 1
Radionuclide: Nonlabeled compound
Specific Activity: -
Analyte/Assay: Unchanged compound (mcg/mL)/HPLC
Sampling time: 10 min, 1, 4, 8, 24, 48, 96, and 168 hr
	C1hr		Last time point
Tissues/organs	conc.	T/P1)	conc.	T/P1)	Time	AUC	t1/2
Heart	1.4	0.08	0.44	22	48	57.3	37.3
Liver	4.5	6	1.85	92.5	48	290	51.7
Kidney	2.8	0.20	1.07	53.5	48	126	36.3
Spleen	6.5	8.6	3.5	175	48	410	46.9
Additional information:

¹⁾ [Tissue]/[Plasma]

EXAMPLE

2.6.5.6 Pharmacokinetics: Plasma Protein Binding Test Article: Curitol Sodium

Study system: In vitro
Target entity, Test system and method: Plasma, Ultrafiltration
Species	Conc. tested	% Bound		Study No.	Location in CTD Vol. Page
Rat	1 - 100uM	82.1 - 85.4		95301	21	150
Dog	1 - 100uM	83.5 - 88.2		95301	21	150
Human	1 - 100uM	75.2 - 79.4		96-103-03	45	1
Additional Information: