Glybera

“All toxicity studies were performed in wild type mice in accordance with GLP.”

“The use of a single species for toxicity evaluation is in accordance with the guideline for gene therapy medicinal products EMEA/CHMP/GTWP/587488/2007 and the guideline on the non-clinical studies required before first clinical use of gene therapy medicinal products (EMEA/CHMP/GTWP/12459/2006).”

“Studies were conducted with AMT-010 (plasmid derived) and several different batches of AMT-011 (baculovirus derived) and doses ranged from 1x1011 – 1x1013 gc/kg.”

“All toxicity studies were performed in wild type mice in accordance with GLP.”

“Reproductive toxicity was studied in pregnant mice in whom no effects of female reproduction or fetal development were detected.”

“Carcinogenicity studies have not been conducted. The applicant argues that the studies conducted to assess integration are sufficient to address risk of carcinogenicity.”

“Studies were conducted with AMT-010 (plasmid derived) and several different batches of AMT-011 (baculovirus derived) and doses ranged from 1x1011 – 1x1013 gc/kg.”

“Moreover, the long-term expression of the therapeutic gene has been taken into account by including an extended follow-up period of 6 months in one of the single dose toxicity studies performed with AMT-011.”

“The non-clinical toxicology programme of Glybera included one single dose toxicity study performed with AMT-010 with a 90-day observation period, two single dose toxicity studies performed with AMT-011 with a 180-day and 105-day observation period, respectively, and a combined reproductive toxicology and breeding study performed with AMT-011.”

“The single dose toxicity studies reflect the clinical dosing situation by addressing the toxic effects following either IM (clinical use) or IV (worst case scenario) administration of AMT-011 at a single occasion.”

“Standard genotoxicity studies, as applied to a conventional chemical drug, are not relevant for this type of product and were not performed.”

“Carcinogenicity studies have not been conducted. The applicant argues that the studies conducted to assess integration are sufficient to address risk of carcinogenicity.”

“Integration analysis did not identify any risky integration hotspots or clonal skewing that might indicate risk of mutagenesis on insertion of AMT-011 into the genome. Integration events were detectable (<2%) but were close to random in the genome.”

“Reproductive toxicity was studied in pregnant mice in whom no effects of female reproduction or fetal development were detected.”

“Standard genotoxicity studies, as applied to a conventional chemical drug, are not relevant for this type of product and were not performed.”

“Standard genotoxicity studies, as applied to a conventional chemical drug, are not relevant for this type of product and were not performed.”

“Standard genotoxicity studies, as applied to a conventional chemical drug, are not relevant for this type of product and were not performed.”

“Studies using techniques to selectively digest genomic DNA showed that the majority (97%) of AMT-011 is not integrated into the genome, implying their presence episomally.”

“In vivo carcinogenicity studies have not been conducted. The applicant argued that there is no existing study design that would be accepted as relevant to assess insertional mutagenesis and the risk of oncogenicity”

“Activity was demonstrated in the dose range 1011-1013 gc/kg compared to the proposed human dose of 1x1012 gc/kg.”

“At the injection sites, histopathological findings including myodegenerative changes and sub-acute inflammation were consistently found in all three single dose toxicity studies upon IM administration of either 1x 1012 or 1 x1013 gc/kg AMT-010 and AMT-011, respectively.”

“on readministration, the second dose was not able to elicit a response, an effect attributable to formation of neutralising antibodies to AAV1 capsid.”

“The main tissues where vector DNA was detected were the injected muscle, liver, spleen and inguinal lymph nodes.”

“The consensus view is that rAAV vector genomes remain episomal with minimal integration into the mammalian genome with no association with oncogenicity.”

“on readministration, the second dose was not able to elicit a response, an effect attributable to formation of neutralising antibodies to AAV1 capsid.”

“High rates of immune response in terms of antibody development were seen in all cases thereby precluding retreatment.”

“Activity was demonstrated in the dose range 1011-1013 gc/kg compared to the proposed human dose of 1x1012 gc/kg.”

“Repeated dose toxicity studies were not conducted as Glybera will be administered once at multiple sites intramuscularly.”

“The non-clinical toxicology programme of Glybera included one single dose toxicity study performed with AMT-010 with a 90-day observation period, two single dose toxicity studies performed with AMT-011 with a 180-day and 105-day observation period, respectively, and a combined reproductive toxicology and breeding study performed with AMT-011.”

“Reproductive toxicity was studied in pregnant mice in whom no effects of female reproduction or fetal development were detected.”

“Reproductive toxicity was studied in pregnant mice in whom no effects of female reproduction or fetal development were detected.”

“Carcinogenicity studies have not been conducted. The applicant argues that the studies conducted to assess integration are sufficient to address risk of carcinogenicity.”

“Studies using techniques to selectively digest genomic DNA showed that the majority (97%) of AMT-011 is not integrated into the genome, implying their presence episomally.”

“In vivo carcinogenicity studies have not been conducted. The applicant argued that there is no existing study design that would be accepted as relevant to assess insertional mutagenesis and the risk of oncogenicity”

“There appears to be progressive and dose-related muscle toxicity at the recommended clinical dose. This has been shown to be reversible.”

“At the injection sites, histopathological findings including myodegenerative changes and sub-acute inflammation were consistently found in all three single dose toxicity studies upon IM administration of either 1x 1012 or 1 x1013 gc/kg AMT-010 and AMT-011, respectively.”

“Overall, the applicant showed relevant pharmacodynamic activity of AMT-010 and AMT-011 in LPL-/- mice and/or cats.”

“However, LPL-/- mice do not develop acute pancreatitis, which is the most severe complication observed in humans suffering from LPLD.”

“A single dose was able to induce a long-lasting expression of transgene, human LPL, which was associated with a clear effect to reduce raised triglycerides very markedly. This is considered to be an acceptable pharmacological surrogate for efficacy.”

“Different batches of AMT-010 (plasmid derived) were used and the applicant intended to compare AMT-010 and AMT-011 (baculovirus derived) in non clinical bridging studies.”

“Studies to investigate absorption have not been conducted. The product is given intramuscularly and the expectation is that Glybera expresses the transgene within the muscle.”