Attachments:
`
`”or": W
`To: W:WWW
`Subject:
`FIN: Cinryze stability studies
`Date:
`
`So the fact that it is possible to concentrate SDOU in 1.5ml has only taken decades to be
`determined!
`
`Hopefully this 1.5 ml injection is not going to hurt too much.
`
`Any volunteers?
`
`Colin.
`
`Colin Broom MD MRCP
`Chief Scientific Officer
`ViroPhan‘na Inc
`730 Stockton Dr
`Exton
`PA 19341
`Tel: (610) 321 6205
`Fax: (610) 458 7330
`Email: colin.broom@viropharma.com
`
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`
`From: Johnson, Judy
`Sent: Wednesday. March 11. 2009 2:01 PM
`To: Broom. Colin; Pietrusko. Robert
`Subject: Cinryze stability studies
`
`Colin and Bob.
`
`Here‘s the most recent draft report from Sanquin on the stability studies which seems to suggest
`that 1.5 mL would be the minimum volume for reconstitution of a vial of Cinryze for sc injection.
`When reconstituted in 1 mL. there was some material sticking to the vial walls.
`
`Judy
`
`Page 1 0f 7
`
`CSL EXHIBIT 1051
`
`Page 1 of 7
`
`CSL EXHIBIT 1051
`
`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`
`0 Confidential
`0 For internal use
`
`0 Limited distribution for:
`0 Product registration
`0 Publication
`
`0 Third parties
`
`Product: Cinryze
`No.:
`036 - xxx
`
`Page:
`Date:
`
`1 - 6
`05Mar2009
`
`Subject:
`
`Stability of Cinryze (3 100 Ufmi) in solution
`
`Researcher:
`
`J. Bloem
`
`Summary:
`
`This report describes the investigation into the solubility and stability of reconstituted Cin ryze
`containing 3 100 Ufml stored for up to 24 hours at 20 :i: 2 "C. it can be concluded that Cinryze
`containing the excipients as described in the BLA1 can be solved in 1 — 5 ml of w.f.i within 10 min.
`No loss of activity, degradation or aggregation was detected after storage at room temperature for
`up to 24 hr in concentrations of up to 500 Ulml.
`
`I3-_—
`Author: J. Bloem
`Manager Product Development:
`A.H.L. Koenderman
`
` _
`
`—
`
`Page 2 0f 7
`
`Page 2 of 7
`
`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`2-5
`
`Page
`
`WNNN
`
`036-100:
`
`Contents:
`
`Introduction
`1.
`2. Material and Methods
`3. Results
`4. Conclusions
`
`1.
`
`Introduction
`
`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 C1-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 Uiml and higher concentrations of
`excipients (Table 1). Also, the stability of this concentrated product at room temperature up to 24
`hrs was tested.
`
`2.
`
`Material and Methods
`
`2.1. Material
`
`For the experiments vials of Cin ryze batch 06002L370A were used.
`Specifications of batch OSCUZLSYOA at release were: C1-inhibitor activity 96 Uiml; protein 14.1 9!];
`pH 7.0; sodium106 mmoIIL; citrate 10 mmoIiL; sucrose 62 mmoliL; osmolality 302 mosmolikg;
`valine 17 mmollL; alanine 13 mmoliL; threonine 38 mmolil.
`
`2.2. Methods
`
`The solubility and stability at C1-inhibitor concentrations of 200, 250, 333 and 500 Uiml were
`tested. As a reference the product in a concentration of 100 Uiml was used. Per experiment several
`vials were used to have enough material to analyse the product at 0, 1, 3, 6 and 24 hr after
`dissolution and storage at 20 i 2 ”C.
`The solubility of the C1-inhibitor protein was determined using M066 (QC protocol).
`Subsequently, the vials were sampled at 5 time points, i.e. t=0, t=1h, 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 (@0398), C‘l-inhibitor antigen (@Oxxx) and HPSEC (PO
`method). 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-inhibitor 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, showing that the dissolution procedure must be done carefuiiy. The
`
`Page 3 of 7
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`Page 3 of 7
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`
`
`036-xxx 3-6
`
`Table 1; Theoretical concentrations of exclplents in Cinryze in a smaller dissolution volume than
`described in the BLA
`C1-
`citrate
`L—valin e
`L-aiani ne
`L~
`Su cross
`Inhibitor
`threonine
`activity
`
`
`
`
`
`
`mmollL
`mmollL
`mmoilL
`mmollL
`mmollL
`%
`
`
`
`1 D
`
`10
`20
`25
`33
`50
`
`17
`34
`43
`5?
`85
`
`13
`26
`33
`43
`65
`
`38
`76
`95
`1 27
`1 90
`
`60
`120
`1 50
`200
`300
`
`2
`4
`5
`7
`
`Table 2: Solubility and appearance of the solution after storage for up to 24 hrs at room
`temperature;
`conform release: no precipitate, no "flakes”. not opalescent
`
`Appearance
`Solubility
`at 3hrs at 24 hrs
`
`
`
`conform
`
`conform
`conform
`conform
`conform
`conform
`
`conform
`conform
`conform
`conform
`
`++: within 5 min.
`+ : 5 min.
`-
`: 5 min., but some material sticked to the wall and did not dissolve completely.
`
`stability of 01 -inhibitor activity after reconstitution was determined at t=0 h, t=1 h. t=3 h t=6 h and t=
`24 h (Table 2) and no loss of C‘l-inhibitor activity was observed after storage at room temperature
`for up to 24 hrs. For the 200 Ulml experiments the measured C1-inhibitor concentration was lower
`than calculated, but the Cl -inhibitor activity did not change after storage. it was decided to repeat
`the experiment with this concentraion and the results will be reported as an amendment to this
`report.
`Also, an 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.
`De analyses zrj‘n klaar maar r'k heb nu green mogelfikheid cm 0'6 figuren re maken. Zal moeten
`wachten tot Judith weer temg is.
`
`4
`
`Conclusions
`
`it can be concluded that dissolving the regular product in less w.f.i. to obtain a presentation
`containing up to 500 Lilml does not affect the stability of the product after reconstitution. However,
`due to the high concentration of the excipients more experiments are needed to find a formulation
`containing a high concentration of C1-inhibitor at lower concentrations of excipients.
`
`Page 4 0f 7
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`Page 4 of 7
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036 - xxx
`
`Page 5 0f 7
`
`Page 5 of 7
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`

`

`SANQUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036-xxx
`
`5—6
`
`Table 3: Cf-inhibitor acthtity after dissolution and storage for up to 24 hrs at room temperature
`
`Cinryze
`
`Activity
`
`
`Ulml
`T = 0
`
`Yield (”/o)
`Specific
`Antigen
`
`
`
`
`activity
`
`
`
`theoretical
`aiL
`Uim .
`
`
`
`102
`90
`94
`97
`
`94
`90
`83
`95
`
`5 ml; 100 Ufinl
`T = 0
`T = 1
`T = 3
`T = 6
`T = 24
`2.5 ml; 200 U/ml
`T = 0
`T = 1
`T = 3
`T = 6
`T = 24
`2 ml; 250 Uflni
`T 2 0
`T = 1
`T = 3
`T = 6
`T = 24
`1.5 mi; 333 U/ml
`T = 0
`T = 1
`T = 3
`T = 6
`T = 24
`1 mi; 500 U/ml
`410
`T = 0
`410
`T = 1
`410
`T = 3
`430
`T = 6
`430
`T = 24
`
`
`105
`
`95
`100
`95
`100
`
`100
`104
`104
`100
`
`100
`100
`105
`
`89
`91
`80
`84
`86
`
`127
`120
`114
`105
`121
`
`220
`210
`220
`210
`220
`
`230
`280
`290
`290
`280
`
`Page 6 0f 7
`
`Page 6 of 7
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`

`

`SANGUIN PLASMA PRODUCTS
`
`Research Report Department of Product Development
`
`036-xxx
`
`6-6
`
`
`
`
`
`
`
`Figure 1: Stability of Cinryze after dissolution at high concentrations.
`
`Figure 2: HPSEC analysis of Cinryze 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.
`HPSEC overlay picture will be added: there were no changes during storage.
`
`Figure 3: HPSEC analysis of Cinryze after dissolution in 2. 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.
`HPSEC overlay picture will be added: there were no changes during storage.
`
`Figure 4: HPSEC analysis of Cinryze alter dissolution in 2 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.
`HPSEC overlay picture will be added: there were no changes during storage.
`
`Figure 5: HPSEC analysis of Cinryze after dissolution in ‘i .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.
`HPSEC overlay picture will be added: there were no changes during storage.
`
`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.
`HPSEC overlay picture will be added: there were no changes during storage.
`
`Page 7 0f 7
`
`Page 7 of 7
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`