Alofisel
darvadstrocel
Sources:EMA assessment report ↗
Species
RatMouse
Longest study
26 wk
Genotox
Not conducted Not conducted
Carcinogenicity
Not conducted Not conducted
Repro
Not conducted Not conducted
First dose
Not conducted Not stated publicly
Speciesopen answer page →
Species
Immunocompromised athymic nude rats (GLP); nude mice for tumourigenicity
“The applicant has chosen immunocompromised athymic rats to perform the biodistribution, toxicology and tumourigenicity studies.”
Why that species
Athymic rats minimise rejection of human cells and extend persistence; immunocompetent rats rejected the cells
“in order to minimize the immune recognition and potential rejection of human eASC in the experimental animal models and to extend Cx601 persistence”
Surrogate molecule
No surrogate; human eASC tested in immunodeficient rats and mice (xenogeneic)
“Human eASC preparations from six donors (3 men and 3 women) were expanded to population doubling levels equal to levels of product for clinical use.”
Species: general tox
Athymic nude rat (perianal; 3- and 6-month post-dose observation) plus IV study
“Two repeat-dose toxicity studies were conducted with human eASC in male and female athymic nude rats (Crl:NIH-Foxn1rnu) via the perianal route, a study with a 3-month post-dose observation period and a study with a 6-month post-dose observation period.”
Species: DART
Not conducted; biodistribution showed no migration to reproductive organs
“Reproductive and developmental toxicity studies have not been performed for Alofisel because preclinical biodistribution studies indicated no migration and integration of eASC into reproductive organs following administration of eASC via different routes.”
Species: carcinogenicity
Nude mouse subcutaneous tumourigenicity test (no conventional carcinogenicity study)
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
Surrogate used in
No surrogate used; human eASC tested in immunocompromised athymic rats
“The applicant has chosen immunocompromised athymic rats to perform the biodistribution, toxicology and tumourigenicity studies.”
Study lengthopen answer page →
Longest pivotal study
26 weeks
“Animals were euthanised at 2 or 26 weeks after the final infusion.”
Pivotal GLP studies
GLP athymic rat: 2 doses 2 weeks apart, perianal (up to 12/24 wk post-dose) and IV (26 wk); 26-wk biodistribution
“Two repeat-dose toxicity studies were conducted with human eASC in male and female athymic nude rats (Crl:NIH-Foxn1rnu) via the perianal route”
Route in tox
Perianal + intrarectal (clinical), intravenous (systemic), intravaginal (supportive)
“Several routes of administration were investigated: a combination of perianal + intrarectal administration reflecting the intended clinical route, the intravenous route exemplifying a systemic administration”
Skipped studiesopen answer page →
Genotoxicity
Not conducted; cell-based product does not interact with DNA
“Genotoxicity studies are not applicable for the product, considering the cell-based nature of the product, which precludes interaction directly with DNA or other chromosomal material.”
Carcinogenicity
Not conducted (conventional); in vitro senescence/karyotype/soft agar and mouse tumourigenicity, no tumours
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
Reproductive & developmental
Not conducted; no migration into reproductive organs in biodistribution
“Reproductive and developmental toxicity studies have not been performed for Alofisel because preclinical biodistribution studies indicated no migration and integration of eASC into reproductive organs”
Safety pharmacology
CNS: functional observational battery in rats showed no effect; CV/respiratory not performed
“eASC treatment did not affect any of the neurological parameters evaluated in the Functional Observational Battery Test”
Genotox: Ames
Not conducted (cell-based product does not interact with DNA)
“Genotoxicity studies are not applicable for the product, considering the cell-based nature of the product, which precludes interaction directly with DNA or other chromosomal material.”
Genotox: in vitro mammalian
Not conducted; karyotype normal diploid in expanded cells
“Analysis revealed normal male or female diploid karyotypes.”
Genotox: in vivo
Not conducted (cell-based product does not interact with DNA)
“Genotoxicity studies are not applicable for the product, considering the cell-based nature of the product, which precludes interaction directly with DNA or other chromosomal material.”
Carcinogenicity model
Subcutaneous tumourigenicity in nude mice: no tumours; in vitro senescence, telomerase, c-myc, soft agar negative
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
Carcinogenicity timing
No standard bioassay; in vivo nude mouse tumourigenicity study (no tumours) completed before the marketing application
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
hERG / QT
Not conducted (no cardiovascular/respiratory safety pharmacology)
“Cardiovascular or respiratory safety pharmacology studies were not performed for eASC which was considered acceptable by the CAT.”
First doseopen answer page →
First-in-human starting dose
Not stated in public reviews
Why it's blank
The public FDA and EMA reviews of Alofisel don't say how the first human dose was chosen. That rationale is in the IND, which is confidential, so it is missing for most approved products.
How it's usually set for cell therapies
FDA expects preclinical proof-of-concept and toxicology studies to identify active dose levels and support the choice of starting dose and escalation. The 2013 guidance gives no numeric formula or scaling rule for the starting dose.
“Selection of potential starting dose level, dose-escalation schedule, and dosing regimen for clinical trials.”
✓ VerifiedFDA 2013: Preclinical Assessment of Investigational Cellular and Gene Therapy…, p. 7Source ↗
This product's starting point
NOAEL / key findings: Perianal doses up to 2.5x10^6 cells/rat well tolerated; IV 10x10^6 cells caused fatal pulmonary embolism (EMA assessment report, p. 23)
NOAEL / key findings
Perianal doses up to 2.5x10^6 cells/rat well tolerated; IV 10x10^6 cells caused fatal pulmonary embolism
“Taken together, two administrations of eASC two weeks apart at doses up to 2.5 x 106 cells were well tolerated in athymic nude rats when administered via the perianal route.”
Risksopen answer page →
Immune & cytokine safety
Low T-cell recognition and delayed NK elimination in vitro; immunocompetent rats showed rejection-type inflammation
“Data generated in other toxicity studies show low potential for T-cell mediated eASC recognition, in vitro and delayed NK cell mediated elimination.”
Biodistribution & persistence
Rats: eASC persisted short term (day 14), restricted to rectum/jejunum locally; lungs after IV
“In conclusion, when administered locally human eASC is only persistent for a short period of time in rats.”
Modality-specific risk
Ectopic tissue formation and tumourigenicity (RMP potential risk); pulmonary embolism if systemic
“Ectopic tissue formation and tumourigenicity has therefore been included in the RMP as important potential risk and will be followed up with the agreed post-authorisation safety study (PASS).”
Anti-drug antibodies: animals
Not stated in public reviews
Not reported in the public reviews. Reviewers summarise selectively, so this usually means it wasn't discussed publicly, not that it wasn't studied.
Anti-drug antibodies: patients
Donor-specific antibodies generated; no apparent effect on efficacy or safety after single treatment (incidence not given)
“A specific risk associated with Alofisel use is the generation of Donor Specific Antibodies (DSA). Based on the current data, the generation of DSA does not appear to affect efficacy or safety after single treatment.”
Tox packageopen answer page →
Before first-in-human
First-in-human: where
Not stated in public reviews
Not reported in the public reviews. Reviewers summarise selectively, so this usually means it wasn't discussed publicly, not that it wasn't studied.
Repeat-dose tox before first-in-human
Not stated in public reviews
Not reported in the public reviews. Reviewers summarise selectively, so this usually means it wasn't discussed publicly, not that it wasn't studied.
First-in-human starting dose
Not stated in public reviews
Why it's blank
The public FDA and EMA reviews of Alofisel don't say how the first human dose was chosen. That rationale is in the IND, which is confidential, so it is missing for most approved products.
How it's usually set for cell therapies
FDA expects preclinical proof-of-concept and toxicology studies to identify active dose levels and support the choice of starting dose and escalation. The 2013 guidance gives no numeric formula or scaling rule for the starting dose.
“Selection of potential starting dose level, dose-escalation schedule, and dosing regimen for clinical trials.”
✓ VerifiedFDA 2013: Preclinical Assessment of Investigational Cellular and Gene Therapy…, p. 7Source ↗
This product's starting point
NOAEL / key findings: Perianal doses up to 2.5x10^6 cells/rat well tolerated; IV 10x10^6 cells caused fatal pulmonary embolism (EMA assessment report, p. 23)
Longest repeat-dose systemic studies
Longest repeat-dose study and recovery
Athymic nude rat repeat-dose study (perianal) with 6-month post-dose observation; IV study similar
“Two repeat-dose toxicity studies were conducted with human eASC in male and female athymic nude rats (Crl:NIH-Foxn1rnu) via the perianal route, a study with a 3-month post-dose observation period and a study with a 6-month post-dose observation period.”
Pivotal GLP studies
GLP athymic rat: 2 doses 2 weeks apart, perianal (up to 12/24 wk post-dose) and IV (26 wk); 26-wk biodistribution
“Two repeat-dose toxicity studies were conducted with human eASC in male and female athymic nude rats (Crl:NIH-Foxn1rnu) via the perianal route”
DART package
Species: DART
Not conducted; biodistribution showed no migration to reproductive organs
“Reproductive and developmental toxicity studies have not been performed for Alofisel because preclinical biodistribution studies indicated no migration and integration of eASC into reproductive organs following administration of eASC via different routes.”
Reproductive & developmental
Not conducted; no migration into reproductive organs in biodistribution
“Reproductive and developmental toxicity studies have not been performed for Alofisel because preclinical biodistribution studies indicated no migration and integration of eASC into reproductive organs”
PPND / ePPND: when
Not conducted; biodistribution showed no migration to reproductive organs
“Reproductive and developmental toxicity studies have not been performed for Alofisel because preclinical biodistribution studies indicated no migration and integration of eASC into reproductive organs following administration of eASC via different routes.”
Carcinogenicity
Species: carcinogenicity
Nude mouse subcutaneous tumourigenicity test (no conventional carcinogenicity study)
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
Carcinogenicity model
Subcutaneous tumourigenicity in nude mice: no tumours; in vitro senescence, telomerase, c-myc, soft agar negative
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
Carcinogenicity timing
No standard bioassay; in vivo nude mouse tumourigenicity study (no tumours) completed before the marketing application
“In in vivo tumourigenicity tests, nude mice administered a subcutaneous injection of eASC at various population doubling levels, equal to levels of product for clinical use, all survived to scheduled sacrifice and no tumour-formation was observed.”
Safety marginopen answer page →
Safety margin at NOAEL
Not stated in public reviews
Not reported in the public reviews. Reviewers summarise selectively, so this usually means it wasn't discussed publicly, not that it wasn't studied.
NOAEL / key findings
Perianal doses up to 2.5x10^6 cells/rat well tolerated; IV 10x10^6 cells caused fatal pulmonary embolism
“Taken together, two administrations of eASC two weeks apart at doses up to 2.5 x 106 cells were well tolerated in athymic nude rats when administered via the perianal route.”
Efficacy evidenceopen answer page →
Efficacy evidence
TNBS-induced colitis mice: dose-dependent improved survival and body weight; no anal fistula model
“Dose dependent, statistically significant improvements in overall survival and body weight loss were associated with eASC treatment relative to TNBS control treatment.”
Why that model
No anal fistula model used, as agreed in CHMP scientific advice; colitis model used instead
“In vivo, no model for anal fistulas was used to provide proof-of-concept data. This was agreed by the CHMP during the 2011 scientific advice.”
Reviewer's view of the efficacy data
EMA: colitis model provides proof-of-concept for eASC effect in inflamed gut
“Instead, an experimental model of colitis was used to show the effect of eASC in an inflamed gut and the data provide evidence for proof-of-concept in this model.”
Flexibilityopen answer page →
Flexibility accepted
DARTAcceptedBeyond the letter
No reproductive and developmental toxicity studies
Why: Biodistribution showed no migration or integration of eASC into reproductive organs · Pharmacology or kinetics
“Reproductive and developmental toxicity studies have not been performed for Alofisel because preclinical biodistribution studies indicated no migration and integration of eASC into reproductive organs following administration of eASC via different routes. This was considered acceptable by the CAT.”
Safety pharmacologyAcceptedBeyond the letter
No cardiovascular or respiratory safety pharmacology studies; CNS assessed by FOB in athymic rats only
“Cardiovascular or respiratory safety pharmacology studies were not performed for eASC which was considered acceptable by the CAT.”
Efficacy modelAcceptedBeyond the letter
No anal fistula model; TNBS colitis model used for proof-of-concept
Why: Agreed in CHMP scientific advice; colitis model showed effect in an inflamed gut · Other reason
“In vivo, no model for anal fistulas was used to provide proof-of-concept data. This was agreed by the CHMP during the 2011 scientific advice.”
“Instead, an experimental model of colitis was used to show the effect of eASC in an inflamed gut and the data provide evidence for proof-of-concept in this model.”
SpeciesVerdict not stated
Toxicology, biodistribution and tumourigenicity in immunocompromised athymic rats rather than standard species
Why: Minimise immune recognition and rejection of human eASC and extend their persistence
“The applicant has chosen immunocompromised athymic rats to perform the biodistribution, toxicology and tumourigenicity studies. The applicant considers that this was done in order to minimize the immune recognition and potential rejection of human eASC in the experimental animal models and to extend Cx601 persistence”
Similar precedents
EbvalloCell therapy · tabelecleucel · EBV-specific allogeneic T cellsRodent only · 20 wkOmisirgeCell therapy · omidubicel · Expanded cord blood HPCRodent only · 12 wkPizensySmall molecules · lactitol · Osmotic laxativeRodent only · 13 wkStrataGraftCell therapy · allogeneic cultured keratinocytes and dermal fibroblasts in murine collagen · Allogeneic keratinocytes + fibroblastsRodent only · 20 wkZalmoxisCell therapy · allogeneic T cells modified with HSV-TK / dLNGFR (nalotimagene carmaleucel) · HSV-TK suicide-gene T cellsRodent only · 3 wkAbecmaCell therapy · idecabtagene vicleucel · BCMA CAR-TNo relevant species · 12 wk