SANOUIN PLASMA PRODUCTS
`
`Mt! Report Dopartrnont 0! Product MW
`
`0 Confidential
`0 For internal use
`0 Limited distribution tor:
`0 Product registration
`0 Publication
`0 Third parties
`
`Product: Cinryzo
`No.:
`080 - 152
`Page:
`1'- B
`Duo:
`15 May 2009
`
`Subloci:
`
`Stability 01 CW (,2 100 U/rri) in ookrtion
`
`Researcher.
`
`J.Bioom
`
`Summary:
`
`This moon dooorlbeelho invettigatlon lrltottiooolubmymdstnblltyot reconstituted Cinryzo
`mum-tog100Uhriotoredtorupto24homat20:2°c.itoonbooonoiudodttutcinryzo
`mwmammmummmm -5miotw.i.iwithin10min.
`Noiouoiaotivity,dogradnflonoraagrooationwndotectodaflorotorogootmntunpardureior
`uptoztivhoomontrationootuptoSOOU/ni.
`
`
`
`Page 1 of 8
`
`CSL EXHIBIT 1050
`
`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036-152
`
`Contents:
`
`Introduction
`1.
`2. Material and Methods
`3. Results
`4. Conclusions
`
`1.
`
`Introduction
`
`2-8
`
`Page
`
`2
`2
`2
`3
`
`For the development of a C1-inhibitor product for subcutaneous administration a more concentrated
`product is preferred. As a first experiment it was tried to dissolve the current registered Cinryze
`product in a smaller volume to obtain a product with a higher Cl-inhibitor concentration but also
`higher excipient concentrations. It was tested if the current product can be dissolved in 2.5, 2, 1.5
`or 1 ml to obtain product containing 200, 250, 333 or 500 Ulml and higher concentrations of
`excipients (Table 1). Also, the stability of this concentrated product at room temperature for up to
`24 hrs was tested.
`
`2.
`
`Material and Methods
`
`2.1. Material
`
`For the experiments vials of Cinryze batch 06002L370A were used.
`Specifications of batch 06002L370A at release were: Cl-inhibitor activity 96 Ulml; protein 14.1 g/l;
`pH 7.0; sodium 106 mmol/L; citrate 10 mmol/L; sucrose 62 mmol/L; osmolality 302 mosmollkg;
`valine 17 mmol/L; alanine 13 mmol/L; threonine 38 mmolll.
`
`2.2. Methods
`
`The solubility and stability at C1-inhibitor concentrations of 200, 250, 333 and 500 Ulml were
`tested. As a reference the product in a concentration of 100 Ulml was used. Per experiment several
`vials were used to have enough material to analyse the product at O, 1, 3, 6 and 24 hr after
`dissolution and storage at 20 1 2 °C.
`The solubility of the C1-lnhibitor protein was determined using SOP M066 (QC protocol).
`Subsequently. the vials were sampled at 5 time points, 1.6. t=0, t=1 h, t=3h, t=6h and t=24h. The
`samples were transferred to Sarstedt tubes, frozen at -30 °C and despatched for the following
`determinations: C1-inhibitor activity (SOP @0398). C1 -inhibitor antigen (SOP @2587) and HPSEC
`(SOP M916). It must be noted that the C1-inhibitor activity assay is not validated for products with
`high concentrations of C1-inhibitor and excipients (a.o. dilution effects).
`
`3.
`
`Results
`
`3.1
`
`Stability of C1-inhlbitor in solution
`
`Cinryze can be dissolved in 1 - 5 ml within 10 min. (Table 2). Only if 1 ml of w.f.i. is used, some
`material is sticking to the wall due to the small volume.
`
`Page 2 of 8
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036-152
`
`3-8
`
`Table 1: Theoretical concentrations of excipients in Cinryze in a smaller dissolution volume than
`described in the BLA
`
`Sodium
`
`citrate
`
`L-valine
`
`L-alanlne L-threonlne
`
`Sucrose
`
`C1-lnhlbltor
`
`
`activity
`Ulml
`
`
`
`
`mmollL
`
`mmollL
`
`mmollL
`
`mmollL
`
`mmollL
`
`mmol/t.
`
`Table 2 : Solubility and appearance of the solution after storage for up to 24 hrs at room
`temperature; conform release: no precipitate, no “flakes"
`
`m Solubility
`
`at 3hrs
`
`Appearance
`
`at 24 hrs
`
`M: within 1 min.
`+ : 5 mln.
`-
`: within 10 mln., but some material sticker! to the wall and did not dissolve completely due to the small volume.
`
`The stability of C1-inhibitor activity after reconstitution was determined at t0 h t=1 h. t=3 h t=6 h
`and t= 24 h (Table 2) and no loss of C1 -inhibitor activity was observed after storage at room
`temperature for up to 24 hrs (Table 3; Figure 1). For the 200 Ulml experiments the measured C1-
`inhibitor concentration was lower than calculated, but the C1-inhibitor activity did not change after
`storage. It was decided to repeat the experiment with the 100. 200 and 500 Ulml fonnulatlons and
`the results are presented in Table 4. For the 200 Ulml experiments the measured C1-inhibitor
`concentration was as expected and thus higher than in the first experiment . However, in this
`experiment the Clinhibitor concentration in the 500 Ulml formulation was lower than calculated.
`The osmolality and pH of the samples were determined (Table 5).
`Also, HPSEC analysis was performed showing that there was no aggregation or degradation after
`storage for up to 24 hrs, even for the highest concentration, 500 Ulml. tested (Figure 2 to 6).
`
`4
`
`Conclusions
`
`it can be concluded that dissolving the regular product in less w.f.i. to obtain a presentation
`containing up to 500 Ulml does not affect the stability of the product after reconstitution. However.
`due to the high concentration of the exclplents more experiments are needed to find a formulation
`containing a high concentration of C1-inhibitor at lower concentrations of excipients.
`
`Page 3 of 8
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036 — 152
`
`4 - 8
`
`Table 3: C1-inhibitor activity after dissolution and storage for up to 24 hrs at room temperature
`
`Activity
`
`U/ml
`
`Yield
`(%)
`
`U/mlé
`
`Antlgen
`
`theoretical
`
`3 L
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`Page 4 of 8
`
`44444llllIIIIIInoun-sob
`
`N 0!44444544444-IIIIIIII"allllllMow-Iownmw-xoA85
`
`{)2o
`
`2ml;
`
`T T T T T
`
`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036-152
`
`5-8
`
`Table 4: C1 -inhibilor activity after dissolution and storage for up to 24 hrs at room temperature;
`second experiment
`
`
`Activity
`
`U/ml
`
`Yield
`(%)
`
`Antigen
`
`theoretical
`
`. IL
`
`Ratio
`Activitylantigen
`U/m .
`
`Q 0
`
`IIIIII:3IIIIIIIINOD—KONQJO«h-8&-
`OIIIIIIII“III
`NOD—‘0
`
`T T T T
`
`. m
`
`T T T T
`
`.5 55
`
`4444
`
`.5
`1 ml,’ 500 U/ml
`
`Table 5: Osmolality and pH at t=0
`
`—-“mosmollk
`
`5 ml; 100 U/ml
`
`2.5 ml: 200 U/ml
`
`2 ml; 250 U/ml
`
`1.5 m1; 333 U/ml
`
`'pH ls determined with pH paper due to small volume left
`
`Page 5 of 8
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`6 - 8
`
`—e— '100 Ulml'
`
`4— '200 U/ml'
`
`+ '250 Ulml'
`
`-u-— '333 Ulml'
`
`+ '500 U/ml'
`
`Figure 1: Stability of Cimyze after dissolution at high concentrations.
`
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`Fleure 2: HPSEC analysis of Cirrryze alter dissolution in 5 ml and storage for up to 24 h at room
`temperature.
`Samples were taken after storage for 10 min, 1 h, 3 h. 6 h and 24 h.
`
`Page 6 of 8
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036 - 152
`
`7 - 8
`
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`Figure 3: HPSEC analysis of Clnryze alter dissolution in 2.5 ml and storage for up to 24 h at room
`temperature.
`Samples were taken after storage for 10 min, 3 h, 6 h and 24 h. The highest peaks
`(black) are the t=0 sample where accidentally a double concentration was analysed.
`
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`Figure 4: HPSEC analysis of Clnryze alter dissolution in 2 ml and storage for up to 24 h at room
`temperature.
`Samples were taken alter storage for 10 mln. 1 h, 3 h. 6 h and 24 h.
`
`Page 7 of 8
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
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`Figure 5: HPSEC analysis of Cinryze after dissolution in 1.5 ml and storage for up to 24 h at room
`temperature.
`Samples were taken after storage for 10 min, 1 h. 3 h, 6 h and 24 h.
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`Figure 6: HPSEC analysis of Cinryze after dissolution in 1 ml and storage for up to 24 h at room
`temperature.
`Samples were taken after storage for 10 min, 1 h, 3 h, 6 h and 24 h.
`
`Page 8 of 8
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