SCIENTIFIC DISCUSSION
`
`Introduction
`1.
`Age-related macular degeneration
`Age-related macular degeneration (AMD) is a disease characterized by progressive degenerative
`abnormalities in the macula, a small area in the central portion of the retina with the highest visual
`acuity.
`AMD is the major cause of vision loss in the elderly population in the Western world. Although the
`disease rarely results in complete blindness and peripheral vision may remain unaffected, central
`vision is gradually blurred, severely affecting ordinary daily activities.
`AMD is characteristically a disease occurring in older patients. Population-based epidemiologic
`studies have provided estimates of prevalence and incidence of AMD among various racial/ethnic
`groups around the world and have shown that AMD is rare before 55 years of age, that it is more
`common in persons 75 years of age or older, and that it is less common in blacks than whites.
`AMD is classified as two different types: the non-exudative (or dry) form and the exudative (or wet)
`form of the disease. The dry from is the most prevalent, accounting for 90% of cases of the disease. It
`is not uncommon that the dry form develops into the wet, neo-vascular form of AMD. Exudative
`AMD, the neovascular form of the disease, is responsible for the majority of cases of severe vision
`loss. Exudative AMD is characterized by the formation of a choroidal neovascular network beneath
`the retina (CNV). This neovascular membrane leaks blood and fluid under the retina and eventually
`progresses to scar formation with destruction of the macula and loss of vision. The prevalence of
`exudative AMD in developed countries rises exponentially with age, with near absence at age 50 to
`1% at age 70 and 5% at age 80.
`The pathogenesis of CNV formation is poorly understood and involves, among many factors, vascular
`growth factors (including but not limited to VEGF), proteases (serine- and metallo-proteases and their
`inhibitors) and inflammation (inflammatory cells, bone marrow-derived progenitors, chemokines).
`There is no curative treatment for AMD. Treatment available includes low vision rehabilitation. A
`certain percentage of patients with exudative AMD can benefit from laser treatment with traditional
`photocoagulation laser or photodynamic therapy (PDT). Photodynamic therapy with verteporfin
`(Visudyne™) has been approved for the treatment of predominantly classic or occult subfoveal
`exudative (neovascular) AMD on the basis of decreased visual deterioration in treated patients
`compared to controls. AMD is currently an area with a need for more efficacious treatments and for
`treatments applicable to all forms of neovascular AMD.
`
`Pegaptanib
`Pegaptanib belongs to the pharmacotherapeutic group of "other ophthalmologicals", ATC code:
`01XA17. Pegaptanib is a pegylated modified oligonucleotide that binds with high specificity and
`affinity to extracellular Vascular Endothelial Growth Factor (VEGF165) inhibiting its activity. VEGF is
`a secreted protein that induces angiogenesis, vascular permeability and inflammation, all of which are
`thought to contribute to the progression of the neovascular (wet) form of AMD. VEGF165 is the VEGF
`isoform preferentially involved in pathological ocular neovascularisation. The selective inhibition in
`animals with pegaptanib proved as effective at suppressing pathological neovascularisation as pan-
`VEGF inhibition, however pegaptanib spared the normal vasculature whereas pan-VEGF inhibition
`did not.
`To increase the intravitreal residence time, a 40 kD branched polyethylene glycol (PEG) molecule has
`been conjugated to the oligonucleotide. Pegaptanib has the following sequence of nucleotides and
`functional groups:
`
`1/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.001
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`
`
`5’-[40kD]-[HN-(CH2)5O]-pCfpGmpGmpArpArpUfpCfpAmpGmpUfpGmpAmpAm
`pUfpGmpCfpUfpUfpAmpUfpAmpCfpAmpUfpCfpCfpGm3’-p-dT
`
`where
`•
`[40kD] represents the two 20 kD PEG chains.
`•
`[HN-(CH2)5O] represents the amino linker connecting PEG and the
`oligonucleotide via a phosphodiester bond.
`• p represents the negatively charged phosphodiester functional groups that have
`Na+ counter ions.
`• Gm or Am and Cf or Uf and Ar represent 2-methoxy, 2-fluoro and 2-hydroxy
`variants of their respective purines and pyrimidines.
`• C, A, U and G code for cytidylic, adenylic, uridylic and guanylic acids.
`
`Secondary
`structure:
`
`
`
` A
`
` clinical development program to investigate the use of pegaptanib sodium in the treatment of
`exudative AMD began in 1999. The nonclinical program began in 1996 and spans the duration of
`clinical development. The development program has been conducted entirely in male and female over
`50-yr patients with exudative AMD (N= 1210) instead of normal volunteers as the product is delivered
`by an intravitreous injection. The risks of such injections were felt to be inappropriate for healthy
`volunteers.
`GMP inspections were performed at the site of manufacture of the active substance and of the finished
`product manufacturing site. Both sites were found to operate in compliance with EU GMP.
`
`
`Part II: Chemical, pharmaceutical and biological aspects
`2.
`Introduction
`Pegaptanib is a synthetic oligonucleotide. Pharmaceutically, the product is presented as a sterilised
`injection 3.47 mg/ml (based on the oligonucleotide weight). The proposed posology is 0.3 mg, every
`six weeks by means of an intravitreal injection (IVT) into the eye using a single-dose, pre-filled
`syringe. Each syringe contains a nominal delivered volume of 90 µl. The drug product is presented in
`a 1 ml glass barrel syringe sealed with a rubber plunger stopper. The syringe has a fixed needle with a
`rubber needle shield and a rigid plastic outer shield. A plastic syringe plunger and flange adapter are
`also supplied for administration purposes.
`Evidence has suggested a causal role of vascular endothelial growth factor (VEGF) in several diseases
`of the human eye in which neovascularization and increased vascular permeability occur. Pegaptanib
`has been developed to bind and block the activity of extracellular VEGF, specifically the 165-amino-
`acid isoform (VEGF165).
`Active Substance
`The active substance is present as the sodium salt, Pegaptanib Sodium, and is a 28-mer
`oligonucleotide aptamer (L. aptus, to fit; Gk. meros, part or region) covalently linked to two 20-kD
`polyethylene glycol (PEG) chains. Two of the nucleotides are ribonucleotides, one is a deoxy
`ribonucleotide, while the rest of the nucleotides contain modified sugars. The modified sugars confer
`increased resistance towards nuclease degradation of the oligonucleotide. The different nucleotides are
`linked via 5’- to 3’-phosphodiester linkages to yield the 28-mer oligonucleotide. The 3’-end thymidine
`contains a 3’-3-linkage to the penultimate 2’methoxyguanosine. The 5’-end of the oligonucleotide
`contains a lysine residue (pentylaminolinker), whose amino groups serve to attach the two PEG units.
`Pegaptanib sodium is hydrophilic and very soluble in water and soluble in a number of organic
`solvents.
`• Manufacture
`
`
`
`2/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.002
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`The manufacture of pegaptanib sodium includes the following steps
`- Oligonucleotide Synthesis, Purification of nonPEGylated Oligonucleotide, PEGylation Reaction,
`
`PEGylated Oligonucleotide Purification and drying.
`The oligonucleotide is manufactured by solid phase organic synthesis using well-established
`methodology.
`•
`Characterisation
`The drug substance is an aptamer, meaning it is the secondary structure (i.e. folding) of the
`oligonucleotide chain that governs the structure which is required for its effect on binding to VEGF165.
`The folding is in turn governed by the primary structure (i.e. nucleotide sequence); hence structural
`characterisation of drug substance involves determination of primary as well as secondary structure.
`The methodologies applied for structural characterisation are well covered. The molecular sequence
`for the pegaptanib sodium molecule was established through a combination of spectroscopic,
`physicochemical and biological techniques. The impurities present in oligonucleotides have been
`investigated.
`•
`Specification
`A variety of tests have confirmed the qualitative and quantitative characteristics of the active
`substance by means of a combination of relevant physicochemical and biological methods.
`Batch analyses indicate satisfactory compliance with the agreed specification and uniformity from
`batch to batch.
`•
`Stability
`The active substance is stored in sealed glass vials. Validated stability-indicating methods have been
`developed and stress studies have demonstrated that it is prone to degradation from light, oxidation
`and heat (40oC/75%RH). Formal studies according to ICH guidelines have been performed at –20 °C
`(± 5 °C), recommended storage conditions, and 5 °C (± 3 °C), accelerated storage conditions, for three
`batches. Supportive data for three batches stored at recommended and accelerated storage conditions,
`up to 24 months is also presented. The results confirm the retest period and storage conditions of the
`active substance.
`
`Medicinal Product
`•
`Pharmaceutical Development
`Aptamers bind with high specificity and affinity to target molecules, including proteins, and as
`expected, the binding relies on the specific three-dimensional conformation of the properly folded
`aptamer. In order to prolong activity at the site of action, the sugar backbone of pegaptanib was
`modified to prevent degradation by endogenous endonucleases and exonucleases, and the polyethylene
`glycol moieties were added to increase the half-life of the drug in the vitreous humour.
`A sterile aqueous parenteral solution was developed as a rational presentation for this product.
`Compatability studies have demonstrate that monobasic sodium phosphate monohydrate, dibasic
`sodium phosphate heptahydrate, sodium chloride, hydrochloric acid and sodium hydroxide, at the
`concentrations used in the formulation, are all compatible with pegaptanib sodium in solution.
`Due to the instability of the active substance to terminal sterilisation , an aseptic, filter sterilisation
`process has been developed.
`• Manufacture of the Product
`Macugen is manufactured by dissolving pegaptanib sodium into a physiologically-compatible
`solution. This is followed by pH adjustment, assay, and dilution to the desired strength. The solution
`is sterilized by filtration and filled into syringes under aseptic conditions. The syringes are labelled
`and then packaged into a foil pouch. The process uses conventional equipment and facilities. There
`are no unduly critical steps and the validation of this standard process is satisfactory.
`
`
`
`3/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.003
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`•
`Product Specification
`The specification of the product is based on that of the active substance with additional pharmaceutical
`tests. Fully characterised reference standards are used in the tests. In addition the specification also
`includes
`tests for delivered volume, osmolality, pH, sterility, endotoxin, and particulate
`contamination..
`Batch analyses indicate satisfactory compliance with the agreed specification and uniformity from
`batch to batch.
`•
`Stability of the Product
`Stability results have been provided for three primary stability lots at the 0.3 mg strength the product
`in the configuration as intended for market. In addition three supportive stability lots at the 1 mg
`strength are presented in the proposed commercial packaging (syringe in foil pouch).
`Tests were performed at 2 – 8oC long term, and 25 oC accelerated, under ICH conditions and the
`characteristics were monitored using stability-indicating methods.
`Results support the shelf-life and storage conditions as defined in the SPC.
`Discussion on chemical, pharmaceutical and biological aspects
`This synthetic peptide has been manufactured characterised and controlled in a way that indicates
`satisfactory purity and uniformity, and the medicinal product has been developed utilising molecular
`development aspects in addition to standard pharmaceutical ones in order to achieve the desired
`clinical effect.
`The finished product is manufactured and tested in a way that indicates a reliable and reproducible
`product in the clinic, throughout the shelflife.
`Macugen should be inspected visually for particulate matter and discoloration prior to administration
`(see SPC section 6.6).
`In the absence of compatibility studies, this medicinal product must not be mixed with other medicinal
`products.
`At the time of the CHMP opinion, a number of minor quality issues related chiefly to method
`development and validation and ongoing stability studies were unresolved; the applicant agreed to
`resolve these as FollowUp Measures within an agreed timeframe.
`
`
`Part III: Toxico-pharmacological aspects
`3.
`Introduction
`The non-clinical pharmacokinetics and toxicity of pegaptanib was evaluated after of both IVT and
`intravenous (IV) administration with the main studies performed in rats, rabbits and dogs. A limited
`reproductive toxicology program was conducted. While no carcinogenicity studies were performed,
`pegaptanib and its potential metabolites, the component’s nucleosides, were evaluated with respect to
`genotoxicity. Most pivotal non-clinical studies were done according to GLP.
`Pharmacology
`Inhibition of VEGF has been shown to, at least partially, prevent neovascularisation in several models,
`both in models using pegaptanib, as those submitted with this application, but also in published studies
`utilising antibodies and small molecules.
`The aptamer of pegaptanib was isolated from RNA libraries containing random nucleotides and
`further modified to obtain high selectivity and affinity (picomolar region) towards especially VEGF165.
`Addition of the PEG-moiety reduced the affinity ~4-fold.
`The pharmacology studies were undertaken to demonstrate the following attributes of pegaptanib
`sodium:
`- High affinity and selectivity for the VEGF165 isoform over VEGF121 isoform.
`- In vitro functional studies to confirm that pegaptanib’s high affinity for this isoform translates to
`
`
`4/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.004
`Regeneron v. Novartis, IPR2021-00816
`
`

`

` VEGF165 antagonism.
`- In vivo studies to demonstrate functional antagonism of VEGF- mediated changes in
`angiogenesis and vascular permeability.
`•
`Primary pharmacodynamics (in vitro/in vivo)
`The pharmacological characterisation of pegaptanib included in vitro studies on pegaptanib binding to
`VEGF165 and related ligands, inhibition of VEGF-binding to its receptors, inhibition of functional
`effects following VEGF165 binding to its receptors (cellular proliferation, calcium flux, and tissue
`factor gene expression), and in vivo effects on vascular permeability and angiogenesis.
`−
`In vitro studies
`The series of in vitro studies that characterized pegaptanib antiangiogenic and anti-permeability
`pharmacology included studies on binding affinity and selectivity of pegaptanib to VEGF165 relative to
`other VEGF isoforms and associated proteins, pegaptanib VEGF165 antagonist activity in human
`umbilical vascular endothelial cell (HUVEC) proliferation and tissue factor expression assays,
`pegaptanib inhibitory effects on VEGF binding to VEGF receptor Fc constructs, and pegaptanib
`inhibition of calcium mobilization in HUVECs.
`Pegaptanib binds in vitro to purified recombinant VEGF165 with high affinity but does not bind to the
`smaller isoform VEGF121 or any of the VEGF-related proteins tested (VEGF-B, VEGF-C, and PlGF).
`Pegaptanib also binds to VEGF188, the murine ortholog of human VEGF189, which is normally bound
`to the cell surface. Binding to VEGF188 was significant, albeit less than obtained with purified
`VEGF165. Pegaptanib effectively inhibits VEGF165 binding to its cellular receptors, as seen with both
`purified constructs of the Flt-1 (VEGFR-1), KDR (VEGFR-2) and NP-1 receptors and on the cell
`surface of cultured human endothelial cells. Pegaptanib sodium has a three-dimensional conformation
`that enables it to bind VEGF165 much like an antibody.
`In addition, VEGF165-induced cellular proliferation, calcium mobilization, and tissue factor gene
`expression are effectively inhibited in cultured human umbilical vein endothelial cells (HUVECs)
`treated with pegaptanib sodium. No investigations have been carried out on choroidal endothelial
`cells. The binding of the aptamer portion of pegaptanib is strong, short-lived and reversible. There is
`no data to show that the sequence does not bind to any off-target ligands/receptors.
`−
`In vivo studies
`In vivo, administration of pegaptanib sodium inhibited hypoxia-induced retinal neovascularization in a
`murine model of retinopathy of prematurity, VEGF-induced corneal angiogenesis in a rat corneal
`pocket model, and dermal vascular leakage in a dermal vascular permeability (modified Miles) assay
`in guinea pigs. Pegaptanib has not been tested in an animal model for choroidal neovascularisation. In
`the hypoxia induced neovascularisation model (retinopathy of prematurity, ROP) in rodents, the key
`role of VEGF in abnormal vascular growth was confirmed. The Applicant has generated additional
`data showing that the intravitreal injections in the ROP model caused a high variability and poor
`recovery of intravitreal concentrations of pegaptanib. Therefore, the i.p. route of administration was
`chosen. In the new studies, ocular neovascularisation could be prevented and a 50% inhibition of
`retinal vascular growth was achieved at 0.21 nM (~1.95 ng/ml). Extrapolation of data indicates that
`levels above the IC50-values are obtained clinically at a 6-week dosing interval if the human vitreous
`T½ exceeds 4 days. Even if the data obtained have their weaknesses, the submitted study is
`reasonable. Further, the Applicant plans a program to evaluate the applicability of 19F-pegaptanib
`MRS to obtain repeated non-invasive measures of pegaptanib levels in the human eye.
`•
`Secondary pharmacodynamics
`Even though pegaptanib is highly selective for VEGF165 compared to closely related targets, no data
`was presented to show that it does not bind to any off-target ligands/receptors, or whether the de-
`pegylated (intact or degraded) aptamer may have antisense properties.
`•
`Safety pharmacology
`in animal models for cardiovascular, respiratory and
`Pegaptanib sodium was evaluated
`neurobehavioral effects, at IV administered doses with associated systemic exposures up to 10-fold
`
`
`5/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.005
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`above the exposures observed after a 3 mg/eye clinical dose. All studies were claimed to be performed
`according to GLP. Pegaptanib did not affect these parameters, nor were any renal parameter changes
`found following up to 3 mg/eye of pegaptanib administered to animals as frequently as every 2 weeks.
`•
`Pharmacodynamic drug interactions
`No pharmacodynamic
`interaction studies were performed. The Applicant claimed that no
`pharmacodynamic drug interactions are expected based on the high specificity of pegaptanib and
`affinity for VEGF165 other than interactions associated with the use of agents with VEGF agonistic or
`anti-agonistic activity. Inhibitors of other anti-angiogenic growth factors as well as anti-VEGF agents
`may enhance the anti-angiogenic activity and some pharmacodynamic actions of pegaptanib when
`given concomitantly. .
`There is a high probability that pegaptanib will be combined with currently applied therapies for AMD
`(e.g. PDT). However, in clinical trials, the concomitant use of PDT and pegaptanib did not seem to
`induce particular safety concerns, nor introduce particular biases. The Applicant has indicated that a
`randomised and double blind clinical study has been initiated to compare the risks and benefits of
`pegaptanib in the presence and absence of PDT treatments.
`
`Pharmacokinetics
`The pharmacokinetics of pegaptanib have been characterised after IV and IVT administration to
`rabbits, dogs and monkeys and after IV administration to mouse.
`
`Analytical procedures were generated to determine pegaptanib in plasma, vitreous fluid and amniotic
`fluid. Pegaptanib was analysed with HPLC (unchanged molecule) and with a dual hybridisation assay
`(DH, oligonucleotide portion of pegaptanib). Additional methods to analyse serum antibodies to
`pegaptanib (ELISA) and 2’-FU (LC-MS/MS) were developed. There is no assay to measure the other
`individual nucleosides. Overall, the analytical assays are considered sufficiently validated, but large
`variations are allowed. The antibody assay has several weaknesses and its usefulness is not clear.
`Besides the lack of positive control, there is no LOQ and it is not known if plasma pegaptanib affects
`the assay, or whether the assay recognises pegaptanib-antibody complexes. Nevertheless, a significant
`antibody response is not expected and difficulties in developing an assay are recognised.
`
`
`Absorption- Bioavailability
`Pegaptanib plasma and vitreous PK were characterized in a broad range of animal species: mice, rat,
`rabbit, dog and monkey. Following IVT administration, pegaptanib was slowly absorbed into the
`systemic circulation. Systemic bioavailability was high in all species and varied between 70 - 100% as
`determined from comparisons of AUC values after IV and IVT administration.
`There were no apparent signs of accumulation of pegaptanib in plasma after repeated dosing to rabbits,
`dogs and monkeys and there were no significant gender differences.
`The plasma terminal half-life of pegaptanib following IVT administration was ~2 to 4 days in animals,
`reflecting the slow exit from the vitreous into systemic circulation.
`After IV administration, plasma pegaptanib increased linearly with dose. Clearance was not affected
`by dose. In a rat study, elimination half lives increased with the weight of the PEG-moiety. In all
`species, the pharmacokinetic profile was similar, even though the half-life of pegaptanib was shorter in
`the dog. The volume of distribution approximately equalled the plasma volume (close to 60 ml/kg),
`suggesting little tissue distribution of pegaptanib.
`Interspecies differences in the pegaptanib plasma pharmacokinetic profile suggest that in the dog
`pegaptanib exits the eye and enters the systemic circulation more rapidly than that seen in other
`species.
`•
`Distribution
`In distribution studies, pegaptanib was labelled with 14C in the lysine residue. After IVT
`administration, radioactivity was mainly located in the vitreous fluid, retina and aqueous humour at 24
`hours post-dose. Over the study duration of 6 weeks, radioactivity remained high in the retina, but
`
` •
`
`
`
`6/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.006
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`diffused particularly to the sclera and choroid, iris and optic nerve. The 15 days half-life of pegaptanib
`related radioactivity in sclera/choroid may support a low frequency posology. A lower increase of
`radioactivity over the study duration was observed in the cornea and also in the lens at the last time
`point, suggesting a diffusion of pegaptanib also to the anterior part of the eye. Compared to ocular
`levels of radioactivity, extraocular tissue levels of radioactivity were very low. After both IVT and IV
`administrations, the highest systemic concentrations of radioactivity (AUC) were obtained in the
`kidney > spleen > bone marrow (vertebra) > lymph node (mesenteric) and liver. The lowest
`concentrations were found in the brain, spinal cord, skeletal muscle (dorsal) and bone (vertebra).
`Following IV administration, low levels of radioactivity were also found in the eye. Generally, after
`IV administration, the tissue to plasma ratios increased over the study interval suggesting that
`radioactivity was cleared faster from plasma than from tissues. No such consistent pattern was seen
`after IVT administration. Pegaptanib does not seem to be associated with red blood cells or melanin.
`Pegaptanib is designed to remain in the eye for an extended period of time and to be cleared from the
`eye by slow passive diffusion into the systemic circulation, from where it should rapidly be cleared.
`Pegaptanib is probably not highly associated with plasma proteins as indicated by its capacity to
`inhibit VEGF induced tissue factor expression both in the presence or absence of plasma proteins.
`Low levels of pegaptanib passes the placenta and approximately 0.04 % of the plasma levels were
`found in amniotic fluid 1 hour (=Tmax) after IV administration of 40 mg/kg pegaptanib to pregnant
`mice.
`
` •
`
`
`Metabolism (in vitro/in vivo)
`In vitro, pegaptanib appears to be metabolized to a greater extent in nuclease and in plasma of lower
`mammal species than in monkey and human plasma. Pegaptanib is metabolized in vivo; the
`component nucleotide, 2’-FU was detected in plasma and urine of DB rabbit following IV and IVT
`pegaptanib administration. Both pegaptanib and 2’-FU are eliminated primarily by the kidney and
`secondarily by the liver.
`Pegaptanib appears to be more metabolically stable in monkeys as the pegaptanib plasma
`concentrations after IVT administration to both eyes are substantially higher in monkeys compared to
`other animals for a given IVT dose.
`In the monkey, it appears that little if any pegaptanib is metabolized in the vitreous. High absolute
`IVT bioavailability of pegaptanib 70% to 100% suggests only a small amount of metabolism is
`occurring in the vitreous of rabbits and dogs. IVT
`•
`Excretion
`Labelled pegaptanib was mainly excreted renally, both intact, as 2’-FU and as unidentified radio-
`activity. After iv. dosing, > 90 % of the dose was recovered in urine. Excretion into bile and passage
`into breast milk were not investigated.
`
`Toxicology
`
`The toxicology of pegaptanib was evaluated in mice, rats, rabbits, dogs and monkeys using the clinical
`route, IVT, and using SC and IV pegaptanib sodium administration. Because pegaptanib was well
`tolerated, and because of limits on pegaptanib sodium dose size administrable to the vitreous chamber,
`no maximum tolerated dose was established.
`
` •
`
`
`Single dose toxicity
`The acute systemic toxicity of pegaptanib sodium was low, with no adverse effects observed at the
`highest dose evaluated (450 mg/kg IV in rats).
`
`Study ID/
`Report No
`
`Species/
`Sex/Number/
`Group
`Rabbits/NZW/ 6m
`
`Monkeys/ Rhesus/ 6 f,
`non-naive
`
`109-97002-T
`
`
`109-98006-B
`(0647-35)
`
`
`Dose/Route
`
`GLP Major findings
`
`0.5 mg in 40 µl, one eye, PBS in
`control eye IVT, ocular exam d. 1,
`5, 12.
`0.5 mg /eye in 67 µl, IVT
`7 or 28 d. observation
`
`No No ocular pegaptanib -
`related effects. Injection-
`related effects in both eyes.
`No None
`
`7/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.007
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`109-980142-T
`(0699-35)
`
`
`109-97003-T
`(804-001)
`109-98011-T
`(0654-35)
`
`Monkeys/ Rhesus
`1/sex/group – cross
`over between lots,
`wash-out 29-35 d.
`
`Rats/SD, 5/sex/group
`
`Monkeys/ Rhesus, 1
`control/sex, 2 m + 1 f
`treated.
`
`1st lot: 0, 0.25, 0.5, 1 mg in 65 µl
`/eye, IVT
`2nd lot: 0.25, 0.5, 1, 1.5, 2 mg in 65
`µl /eye, IVT
`Up to 29 d. observation.
`0, 50, 150, 450 mg/kg, IV bolus
`
`0, 5 mg/kg, IV, 1 hr infusion, 7 d.
`observation
`
`No No ocular pegaptanib-
`related effects. Slight
`injection-related effects in
`control and treated eyes.
`
`Yes None
`
`No None
`
`
`
` •
`
`
`Repeat dose toxicity (with toxicokinetics)
`From the total of studies performed, the 6-month rabbit and 9-month dog studies are considered
`pivotal.
`
`Species/Sex/
`Study ID/
`Number/Group
`Report No
`109-98003-T Rabbits/DB,
`5/sex/group
`
`Dose/Route
`
`Duration
`
`11 weeks
`1 dose/2
`weeks
`(6 doses)
`
`NOAEL
`(drug)
`1 mg/eye
`for 6 doses
`and 2
`mg/eye for
`2 doses
`
`Major findings
`
`Slight to moderate dose-
`dependent presence
`vitreal macrophages and
`mild cyclitis.
`Injection-related ocular
`effects.
`
`Group 1: 0
`Group 2: 0.1
`(first 4 doses)/
`2 (2 doses)
`Group 3: 0.3
`Group 4:1
`mg/eye IVT in
`50 µl
`0, 2 mg in 100
`µl/eye, IVT
`
`No treatment-related
`effects. Injection-related
`ocular effects.
`
`Dose-dependent
`endotoxin-induced ocular
`inflammation after 1st
`dose. Severe at 1 and 2
`mg. (Drug lot 11838.26)
`
`No treatment-related
`effects with Drug lot
`97000690. Injection-
`related ocular effects.
`10 mg/kg: Organ weights
`↑. Trace chronic
`progressive nephropathy
`(m).
`From 1 mg/kg: mild
`lymphoid depletion
`(splenic white pulp) (m),
`vacuolated macrophages
`in several organs.
`No treatment-related
`effects. Injection-related
`ocular effects.
`
`144-002
`Non-GLP
`
`
`Dogs/Beagles,
`control 2f, treated
`1/sex
`
`109-98010-T
`Initial Lot:
`11838.26
`2nd Lot:
`97000690
`
`Monkeys/ Rhesus
`0.5 mg: 4m+2f
`1-2 mg: 4/sex
`
`Naïve animals
`(3/sex/group) were
`assigned to the 0, 0.1
`and 0.25 mg/eye
`groups
`
`Initially: 0, 0.5,
`1, 2 mg in 66
`µl/eye, IVT
`
`After 1st dose:
`0.1 (dose 1-4),
`1 (dose 5-6),
`0.25, 0.5 in 66
`µl/eye, IVT
`
`2 mg/eye
`
`1 mg/eye
`after 2
`doses
`
`3 weeks
`1
`dose/week
`(3 doses)
`3 months
`(LD only)
`1 dose/2
`weeks
`(6 doses)
`
`109-98004-T
`(804-002)
`
`Rats/SD 10/sex/group 0, 0.1, 1, 10
`mg/kg/day, IV
`bolus
`
`13 weeks
`
`1 mg/kg
`
`0460LE15.001
`Pivotal
`
`
`Rabbits/NZW, 7-
`9/sex/group.
`Reversibility 2/sex in
`control and 2 mg
`groups.
`
`0, 0.2, 0.67, 2
`mg in 67 µl/eye
`IVT
`
`2 mg OU
`
`6 months
`with 6 week
`recovery
`1 dose/2
`weeks
`(13 doses)
`
`
`
`8/39
`
`©EMEA 2006
`
`Medicinal product no longer authorised
`
`Novartis Exhibit 2056.008
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`Study ID/
`Report No
`0472DE15.001
`Pivotal
`
`
`Species/Sex/
`Number/Group
`Dogs/Beagle
`5-7/sex/group.
`Reversibility 2/sex in
`control and 3 mg
`groups.
`
`Dose/Route
`
`Duration
`
`0, 0.3, 1, 3 in
`100µl/eye, IVT
`
`Major findings
`
`NOAEL
`(drug)
`3 mg OU Minimal to mild
`lymphocytic infiltration
`in a variety of tissues.
`Vitreal floaters/ strands.
`No other treatment-
`related effects. Injection-
`related ocular effects.
`
`9 months
`with 6 week
`recovery
`1 dose/2
`weeks
`(20 doses)
`
`
`OU= both eyes
`
`In IV dose studies, systemic effects were observed in rats only and were limited to vacuolated cells in
`multiple organs and a mild increase in severity and incidence of chronic progressive nephropathy.
`There were no pegaptanib sodium-related systemic adverse effects observed in IVT repeat-dose
`studies. The vacuolated cells, observed in multiple organs in both male and female rats are considered
`to reflect phagocytosis of the 40 kD PEG moiety of the pegaptanib molecule by macrophages. The
`increase in absolute organ weights noted in this study might have been associated with the vacuolated
`cells. There was no cell damage associated with these vacuoles and this finding was not considered
`adverse. The slight to mild chronic progressive nephropathy was observed in all groups in the repeat
`IV dose rat study including controls; however, in male rats administered 10 mg/kg/day pegaptanib
`sodium IV the incidence and severity were mildly increased as compared to the other groups. In these
`males, a slight decrease in serum protein and albumin was also observed and was likely related to the
`observed chronic progressive nephropathy. The NOAEL for this study, based on this finding, was 1
`mg/kg/day in males. The pegaptanib plasma concentration (30 minutes postdose on Day 91) at the
`NOAEL was approximately 240-fold higher than human maximum pegaptanib plasma concentrations
`following 3 mg/eye IVT doses.
`There were no relevant pegaptanib sodium-related local adverse effects in single- and repeat-dose
`toxicity studies by the IVT route of administration. In the 11-week Dutch-Belted rabbit study there
`was an apparent attenuation of retinal vessels and ocular macrophage infiltrate. The presence of
`retinal vessel attenuation was considered a spurious finding in this study as it was not supported by
`fundus photographic data. Electroretinograms (ERG) appeared normal with the exception of an
`increase in the week 7 latency following single flash with scotopic white light in high-dose animals (1
`mg/eye). In additio