Atlas/Guides

Pre-IND nonclinical readiness checklist

A pre-IND self-check for small biotech teams. Each question maps to a verbatim requirement or recommendation in an ICH guideline, FDA guidance or US regulation, with page number, so you can confirm your nonclinical package supports a first-in-human IND before you file.

0 of 36 checked · 10 areas · every item cites the guidance it comes from

Test article and formulation

Have you documented every chemistry and manufacturing difference between your clinical drug product and the material dosed in your pivotal tox studies?FDA expects the IND to describe these differences and their safety impact, or state there are none; otherwise the tox data may not support the clinical product.
What the guidance says
“sponsors should describe any chemistry and manufacturing differences between the drug product proposed for clinical use and the drug product used in the animal toxicology trials that formed the basis for the sponsor's conclusion that it was safe to proceed with the proposed clinical study.”
✓ VerifiedFDA 1995: Guidance for Industry: Content and Format of Investigational New…, p. 7Source ↗
Is the biologic lot used in your definitive pharmacology and toxicology studies comparable to the product planned for the first clinical study?ICH S6 expects comparability; after a process or formulation change, comparability should be shown (identity, purity, stability, potency) or animal findings may not extrapolate. biologics
What the guidance says
“In general, the product that is used in the definitive pharmacology and toxicology studies should be comparable to the product proposed for the initial clinical studies.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 6Source ↗
For each GLP study, are the dose formulations analysed for uniformity, concentration and stability in the vehicle?21 CFR 58.113 requires analytical testing of every test article mixed with a carrier; without it, the doses actually delivered are unverified.
What the guidance says
“For each test or control article that is mixed with a carrier, tests by appropriate analytical methods shall be conducted: (1) To determine the uniformity of the mixture and to determine, periodically, the concentration of the test or control article in the mixture.”
✓ VerifiedeCFR 2026: 21 CFR Part 58 Good Laboratory Practice for Nonclinical Laboratory…, p. 1Source ↗
Does any excipient lack existing safety data covering your proposed exposure level, duration or route?If so, FDA treats it as a new excipient that needs its own nonclinical safety qualification, which can add studies to the program.
What the guidance says
“are not fully qualified by existing safety data with respect to the currently proposed level of exposure, duration of exposure, or route of administration.”
✓ VerifiedFDA 2005: Guidance for Industry: Nonclinical Studies for the Safety…, p. 4Source ↗

Species selection and relevance

Before first dosing, have you compared in vitro metabolism and plasma protein binding across your tox species and human?ICH M3(R2) expects these data, plus systemic exposure in the tox species, before human trials; they support the species choice and exposure margins. small molecules
What the guidance says
“In vitro metabolic and plasma protein binding data for animals and humans and systemic exposure data (ICH S3A, Ref. 7) in the species used for repeated-dose toxicity studies generally should be evaluated before initiating human clinical trials.”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 11Source ↗
Have you shown that each tox species is pharmacologically relevant, i.e., expresses the target or epitope and responds to your biologic?ICH S6 defines a relevant species by pharmacological activity; this justification is a core part of a biologics tox package. biologics
What the guidance says
“A relevant species is one in which the test material is pharmacologically active due to the expression of the receptor or an epitope (in the case of monoclonal antibodies).”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 7Source ↗
Are you avoiding toxicity studies in species where your biologic is not pharmacologically active?ICH S6 discourages studies in non-relevant species as potentially misleading; if no relevant species exists, transgenic models or homologous proteins should be considered. biologics
What the guidance says
“Toxicity studies in non-relevant species may be misleading and are discouraged.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 8Source ↗
If two relevant species gave similar short-term findings, have you justified running longer-term studies in only one?ICH S6(R1) says one species is usually sufficient for longer-term studies when short-term findings are similar or understood from the mechanism. biologics
What the guidance says
“If the toxicological findings from these studies are similar or the findings are understood from the mechanism of action of the product, then longer-term general toxicity studies in one species are usually considered sufficient.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 15Source ↗

Study duration and design vs the planned clinical trial

Is each repeated-dose tox study at least as long as the longest planned dosing period in your first clinical trial?ICH M3(R2) Table 1 links tox duration to clinical duration; trials longer than the supporting studies are generally not supported. small molecules
What the guidance says
“Clinical trials of longer duration should be supported by repeated-dose toxicity studies of at least equivalent duration.”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 12Source ↗
If your first trial doses for less than 14 days, have you matched the tox study duration to it?ICH M3(R2) Table 1 footnote a allows tox studies of the same duration as a clinical study under 14 days. small molecules
What the guidance says
“Clinical studies of less than 14 days can be supported with toxicity studies of the same duration as the proposed clinical study.”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 12Source ↗
Is the high dose in your general tox studies justified (MTD, exposure saturation, MFD, limit dose, or a 50-fold AUC margin)?ICH M3(R2) section 1.5 sets accepted high-dose limits; an unjustified top dose may leave clinically relevant toxicity uncharacterised. small molecules
What the guidance says
“Doses providing a 50-fold margin of exposure (usually based on group mean AUC values [see Note 1] of the parent drug or the pharmacologically active molecule of a pro-drug) to the clinical systemic exposure generally are also considered acceptable as the maximum dose for acute and repeated-dose toxicity studies in any species.”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 9Source ↗
Do the route and dosing regimen in your biologic tox studies reflect the intended clinical use?ICH S6 expects route and regimen (e.g., daily vs intermittent) to reflect clinical use, with toxicokinetics where feasible. biologics
What the guidance says
“The route and dosing regimen (e.g., daily versus intermittent dosing) should reflect the intended clinical use or exposure.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 11Source ↗

Genotoxicity, carcinogenicity and reproductive toxicity plan

If your first trial uses multiple doses, will a mammalian chromosomal damage assay be complete in addition to the gene mutation assay?ICH M3(R2): a gene mutation assay generally supports single-dose trials; multiple-dose trials also need a chromosomal damage assessment. small molecules
What the guidance says
“To support multiple dose clinical development trials, an additional assessment capable of detecting chromosomal damage in a mammalian system(s) should be completed (Ref. 10).”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 23Source ↗
Have you confirmed carcinogenicity data are not needed at IND, i.e., there is no significant cause for concern?ICH M3(R2) places carcinogenicity studies at the marketing stage; results support clinical trials only where there is significant cause for concern.
What the guidance says
“If carcinogenicity studies are recommended for the clinical indication, they should be conducted to support the marketing application.”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 23Source ↗
If enrolling men in Phase 1, do your repeated-dose tox studies evaluate male reproductive organs?ICH M3(R2) allows men in Phase I and II before a male fertility study because repeated-dose studies evaluate male reproductive organs.
What the guidance says
“Men can be included in Phase I and II trials before the conduct of the male fertility study since an evaluation of the male reproductive organs is performed in the repeated-dose toxicity studies (Note 2).”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 24Source ↗
If enrolling women of childbearing potential without embryo-fetal data, does the protocol include pregnancy-prevention precautions?In the US, embryo-fetal development studies can be deferred until before Phase III only when trials use precautions to prevent pregnancy.
What the guidance says
“In the United States, assessment of embryo-fetal development can be deferred until before Phase III for WOCBP using precautions to prevent pregnancy in clinical trials (see above).”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 25Source ↗

Safety pharmacology

Has the core battery (cardiovascular, respiratory, central nervous system) been evaluated before first human dosing?ICH S7A expects core battery functions investigated before first administration in humans, plus follow-up studies where there is cause for concern.
What the guidance says
“The effects of a test substance on the functions listed in the safety pharmacology core battery should be investigated prior to first administration in humans.”
✓ VerifiedICH 2000: ICH S7A: Safety Pharmacology Studies for Human Pharmaceuticals, p. 11Source ↗
Does your cardiovascular assessment measure blood pressure, heart rate and ECG?ICH S7A names these as cardiovascular core battery endpoints; methods for repolarization and conduction abnormalities should also be considered.
What the guidance says
“Blood pressure, heart rate, and the electrocardiogram should be evaluated.”
✓ VerifiedICH 2000: ICH S7A: Safety Pharmacology Studies for Human Pharmaceuticals, p. 9Source ↗
If your biologic is a novel class or not highly target-specific, have you planned a more extensive safety pharmacology evaluation?ICH S7A: highly target-specific biologics may cover endpoints within tox studies, but novel-class or less specific products warrant more extensive safety pharmacology. biologics
What the guidance says
“For biotechnology-derived products that represent a novel therapeutic class and/or those products that do not achieve highly specific receptor targeting, a more extensive evaluation by safety pharmacology studies should be considered.”
✓ VerifiedICH 2000: ICH S7A: Safety Pharmacology Studies for Human Pharmaceuticals, p. 11Source ↗

First-in-human dose and escalation

Does your starting-dose rationale integrate all nonclinical data: pharmacological dose response, tox profile and pharmacokinetics?ICH M3(R2) expects all relevant nonclinical data, with the NOAEL in the most appropriate species usually most important.
What the guidance says
“All of the relevant nonclinical data, including the pharmacological dose response, the pharmacological/toxicological profile, and pharmacokinetics, should be considered when determining the recommended starting dose in humans.”
✓ VerifiedICH 2009: ICH M3(R2): Guidance on Nonclinical Safety Studies for the Conduct…, p. 14Source ↗
If you did not base the healthy-volunteer starting dose on the most sensitive species, have you justified why?FDA's default is the species with the lowest HED unless species-relevance data (ADME differences, class experience) support another.
What the guidance says
“In the absence of data on species relevance, a default position is that the most appropriate species for deriving the MRSD for a trial in adult healthy volunteers is the most sensitive species (i.e., the species in which the lowest HED can be identified).”
✓ VerifiedFDA 2005: Estimating the Maximum Safe Starting Dose in Initial Clinical…, p. 12Source ↗
If your safety factor differs from 10, especially if lower, have you written out the reasoning?FDA's default safety factor is 10; any departure, particularly a lower one, should be clearly explained.
What the guidance says
“It is incumbent on the evaluator to clearly explain the reasoning behind the applied safety factor when it differs from the default value of 10, particularly if it is less than 10.”
✓ VerifiedFDA 2005: Estimating the Maximum Safe Starting Dose in Initial Clinical…, p. 13Source ↗
Have you compared the NOAEL-based starting dose with the pharmacologically active dose, especially for antibodies or growth factors?For such classes, toxicity may come from exaggerated pharmacology, so the PAD may be the more sensitive limit and warrant a lower starting dose.
What the guidance says
“The PAD in these cases may be a more sensitive indicator of potential toxicity than the NOAEL and might therefore warrant lowering the MRSD.”
✓ VerifiedFDA 2005: Estimating the Maximum Safe Starting Dose in Initial Clinical…, p. 15Source ↗

Modality-specific: biologics

Are you banking samples during biologic tox studies so anti-drug antibodies can be measured if needed?ICH S6(R1): ADA analysis may be needed to interpret altered PD, exposure changes or immune reactions; banked samples allow analysis later. biologics
What the guidance says
“it is often useful to obtain appropriate samples during the course of the study, which can subsequently be analyzed when warranted to aid in interpretation of the study results.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 17Source ↗
Have you omitted the standard small-molecule genotoxicity battery for your protein, unless there is a specific concern (e.g., an organic linker)?ICH S6 says the standard battery is not applicable or needed for biotechnology-derived products; avoid spending on studies with uninterpretable results. biologics
What the guidance says
“The range and type of genotoxicity studies routinely conducted for pharmaceuticals are not applicable to biotechnology-derived pharmaceuticals and therefore are not needed.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 12Source ↗
Is reversibility of effects at clinically relevant exposures addressed, through a non-dosing period or scientific rationale?ICH S6(R1) expects recovery from potentially adverse effects to be understood; complete recovery need not be shown. biologics
What the guidance says
“Recovery from pharmacological and toxicological effects with potential adverse clinical impact should be understood when they occur at clinically relevant exposures.”
✓ VerifiedICH 2011: ICH S6(R1): Preclinical Safety Evaluation of Biotechnology-Derived…, p. 17Source ↗

Modality-specific: oncology (advanced cancer, ICH S9)

For a small-molecule anticancer drug, is the start dose based on 1/10 rodent STD10 or 1/6 non-rodent HNSTD?ICH S9 names these as common approaches; the start dose should still be justified with all nonclinical data. oncology
What the guidance says
“A common approach for many small molecules is to set a start dose at 1/10 the Severely Toxic Dose in 10% of the animals (STD 10) in rodents. If the non-rodent is the most appropriate species, then 1/6 the Highest Non-Severely Toxic Dose (HNSTD) is considered an appropriate starting dose.”
✓ VerifiedICH 2009: ICH S9: Nonclinical Evaluation for Anticancer Pharmaceuticals, p. 13Source ↗
Do your tox studies approximate the planned clinical dosing schedule?ICH S9 notes schedule strongly influences toxicity; a schedule change not supported by existing data usually needs an additional single-species study. oncology
What the guidance says
“As the toxicity of the pharmaceutical can be greatly influenced by its schedule of administration, an approximation of its clinical schedule should be evaluated in toxicology studies.”
✓ VerifiedICH 2009: ICH S9: Nonclinical Evaluation for Anticancer Pharmaceuticals, p. 7Source ↗
Are vital-organ functions covered within general tox (e.g., ECGs in non-rodents) rather than stand-alone safety pharmacology studies?ICH S9 does not call for stand-alone safety pharmacology in advanced cancer unless a specific concern exists. oncology
What the guidance says
“Conducting stand-alone safety pharmacology studies to support studies in patients with advanced cancer is not called for.”
✓ VerifiedICH 2009: ICH S9: Nonclinical Evaluation for Anticancer Pharmaceuticals, p. 7Source ↗

Modality-specific: cell and gene therapy

Are your definitive (IND-enabling) studies run with the same CGT product that patients will receive, or a justified analogue?FDA recommends the clinical product where possible; differences between preclinical and clinical lots should be discussed in the IND. cell and gene therapy
What the guidance says
“When possible, the investigational CGT product that will be administered to the patient population should be used in the definitive preclinical studies.”
✓ VerifiedFDA 2013: Preclinical Assessment of Investigational Cellular and Gene Therapy…, p. 7Source ↗
Do definitive tox studies use the intended clinical delivery device and a route mimicking the clinical one?FDA expects the clinical device in definitive tox studies or a justification; device and procedure risks may need their own data. cell and gene therapy
What the guidance says
“The delivery device intended for use in the clinical studies should be used to administer the investigational CGT product in the definitive toxicology studies; justification should be provided if the intended clinical delivery device is not used.”
✓ VerifiedFDA 2013: Preclinical Assessment of Investigational Cellular and Gene Therapy…, p. 13Source ↗
For a gene therapy, have you run biodistribution studies before first dosing, or justified not doing so?FDA expects biodistribution for new vector classes or significant vector, formulation, route or dose changes, or a written justification. cell and gene therapy
What the guidance says
“Justification should be provided if biodistribution studies are not conducted prior to initiation of early-phase clinical trials.”
✓ VerifiedFDA 2013: Preclinical Assessment of Investigational Cellular and Gene Therapy…, p. 31Source ↗

GLP status and IND content

Does the IND include a GLP compliance statement for each study subject to Part 58, with reasons for any noncompliance?21 CFR 312.23(a)(8)(iii) requires this statement for every nonclinical laboratory study subject to the GLP regulations.
What the guidance says
“For each nonclinical laboratory study subject to the good laboratory practice regulations under part 58, a statement that the study was conducted in compliance with the good laboratory practice regulations in part 58, or, if the study was not conducted in compliance with those regulations, a brief statement of the reason for the noncompliance.”
✓ VerifiedeCFR 2026: 21 CFR 312.23 IND content and format, p. 1Source ↗
Will each pivotal tox study supporting the trial have a full data tabulation ready for FDA review?21 CFR 312.23(a)(8)(ii)(b) requires full tabulated data for each toxicology study intended primarily to support the proposed trial.
What the guidance says
“For each toxicology study that is intended primarily to support the safety of the proposed clinical investigation, a full tabulation of data suitable for detailed review.”
✓ VerifiedeCFR 2026: 21 CFR 312.23 IND content and format, p. 1Source ↗
Have you named the qualified individual(s) who concluded it is reasonably safe to proceed, and where studies and records are located?21 CFR 312.23(a)(8) requires evaluator identity and qualifications plus study and record locations for inspection.
What the guidance says
“Such information is required to include the identification and qualifications of the individuals who evaluated the results of such studies and concluded that it is reasonably safe to begin the proposed investigations and a statement of where the investigations were conducted and where the records are available for inspection.”
✓ VerifiedeCFR 2026: 21 CFR 312.23 IND content and format, p. 1Source ↗
If final audited tox reports won't be ready at filing, is an integrated summary based on unaudited drafts prepared?FDA guidance accepts this route, with final quality-assured reports expected within 120 days of the human study's start.
What the guidance says
“Therefore, if final, fully quality-assured individual study reports are not available at the time of IND submission, an integrated summary report of toxicologic findings based on the unaudited draft toxicologic reports of the completed animal studies may be submitted.”
✓ VerifiedFDA 1995: Guidance for Industry: Content and Format of Investigational New…, p. 13Source ↗

Check your plan against every approval

Score your species, study length and first-dose approach against the precedent in your modality.

Check my plan