`
`Exhibit F
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 2 of 77 PageID #: 379
`
`US011141378B2
`
`( 12 ) United States Patent
`Yaworski et al .
`
`US 11,141,378 B2
`( 10 ) Patent No .:
`( 45 ) Date of Patent :
`* Oct . 12 , 2021
`
`( 72 )
`
`( 54 ) LIPID FORMULATIONS FOR NUCLEIC
`ACID DELIVERY
`( 71 ) Applicant : ARBUTUS BIOPHARMA
`CORPORATION , Vancouver ( CA )
`Inventors : Edward Yaworski , Maple Ridge ( CA ) ;
`Kieu Lam , Surrey ( CA ) ; Lloyd Jeffs ,
`Delta ( CA ) ; Lorne Palmer , Vancouver
`( CA ) ; Ian MacLachlan , Mission ( CA )
`( 73 ) Assignee : ARBUTUS BIOPHARMA
`CORPORATION , Vancouver ( CA )
`Subject to any disclaimer , the term of this
`patent is extended or adjusted under 35
`U.S.C. 154 ( b ) by 0 days .
`This patent is subject to a terminal dis
`claimer .
`( 21 ) Appl . No .: 17 / 227,802
`Apr. 12 , 2021
`( 22 ) Filed :
`( 65 )
`Prior Publication Data
`Sep. 2 , 2021
`US 2021/0267895 A1
`
`( * ) Notice :
`
`( 63 )
`
`Related U.S. Application Data
`Continuation of application No. 17 / 094,724 , filed on
`Nov. 10 , 2020 , which is a continuation of application
`No. 16 / 422,441 ,
`filed on May 24 , 2019 , now
`abandoned , which is a continuation of application No.
`15 / 840,933 , filed on Dec. 13 , 2017 , now abandoned ,
`which is
`continuation of application No.
`a
`15 / 670,742 , filed on Aug. 7 , 2017 , now abandoned ,
`which is
`continuation of application No.
`15 / 164,803 , filed on May 25 , 2016 , now abandoned ,
`which is
`continuation of application No.
`a
`14 / 462,441 , filed on Aug. 18 , 2014 , now Pat . No.
`9,364,435 , which is a continuation of application No.
`13 / 928,309 , filed on Jun . 26 , 2013 , now Pat . No.
`8,822,668 , which is a continuation of application No.
`13 / 253,917 , filed on Oct. 5 , 2011 , now Pat . No.
`8,492,359 , which is a continuation of application No.
`a
`12 / 424,367 , filed on Apr. 15 , 2009 , now Pat . No.
`8,058,069 .
`
`a
`
`2
`
`( 60 ) Provisional application No. 61 / 045,228 , filed on Apr.
`15 , 2008 .
`( 51 ) Int . Cl .
`CO7H 21/02
`CO7H 21/04
`A61K 9/127
`A61K 48/00
`CO7H 21/00
`C07 ) 9/00
`C12N 15/113
`A61K 31/713
`C12N 15/11
`
`( 2006.01 )
`( 2006.01 )
`( 2006.01 )
`( 2006.01 )
`( 2006.01 )
`( 2006.01 )
`( 2010.01 )
`( 2006.01 )
`( 2006.01 )
`
`( 52 ) U.S. CI .
`CPC
`
`A61K 9/1272 ( 2013.01 ) ; A61K 9/1271
`( 2013.01 ) ; A61K 31/713 ( 2013.01 ) ; A61K
`48/0025 ( 2013.01 ) ; C07H 21/00 ( 2013.01 ) ;
`CO7J 9/00 ( 2013.01 ) ; C12N 15/111 ( 2013.01 ) ;
`C12N 15/113 ( 2013.01 ) ; C12N 15/1137
`( 2013.01 ) ; C12N 2310/14 ( 2013.01 ) ; C12N
`2320/32 ( 2013.01 )
`
`( 58 ) Field of Classification Search
`CPC
`C12N 15/113
`See application file for complete search history .
`
`( 56 )
`
`References Cited
`U.S. PATENT DOCUMENTS
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`Primary Examiner Amy H Bowman
`( 74 ) Attorney , Agent , or Firm Kilpatrick Townsend &
`Stockton LLP
`
`ABSTRACT
`( 57 )
`The present invention provides novel , stable lipid particles
`comprising one or more active agents or therapeutic agents ,
`methods of making the lipid particles , and methods of
`delivering and / or administering the lipid particles . More
`particularly , the present invention provides stable nucleic
`acid - lipid particles ( SNALP ) comprising a nucleic acid
`( such as one or more interfering RNA ) , methods of making
`the SNALP , and methods of delivering and / or administering
`the SNALP .
`
`30 Claims , 24 Drawing Sheets
`Specification includes a Sequence Listing .
`
`
`
`( 56 )
`
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`US 11,141,378 B2
`Page 3
`
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`* cited by examiner
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`Sheet 1 of 24
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`US 11,141,378 B2
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`Sample 1 Sample 2
`Sample 5
`Sample 7 Sample 8
`* Sample 3 not find Sample 4
`
`from Sample 6
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`
`
`100
`
`FIG . 1A
`
`10
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`nM Total siRNA
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`Activity of SNALP on HT29 Cell Line
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 7 of 77 PageID #: 384
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`1
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`0.8
`0.6
`1.2 1
`
`) Cells Untreated Viability ( %
`
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`0.2
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`0
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`
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`U.S. Patent
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`Oct. 12. 2021
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`Sheet 2 of 24
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`Sample 13 + Sample 14 Sample 15 Sample 16
`Sample 10 * Sample 11 word in Sample 12
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`the Sample 9
`
`100
`
`FIG . 1B
`
`10
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`
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`nM Total siRNA
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`Activity of SNALP on HT29 Cell Line
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 8 of 77 PageID #: 385
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`1.2 1 0.8
`0.6
`0.2
`0.4
`
`
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`
`
`) Cells Untreated Viability ( %
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`1
`0
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`U.S. Patent
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`Oct. 12. 2021
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`Sheet 3 of 24
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`Activity of SNALP Upon Intravenous Administration in Mice
`group mean SD ( n = 4 )
`
`2.5
`
`Ratio
`mRNA
`
`Liver ApoB : GAPD
`
`2.0
`
`1.5
`
`10
`
`0.5
`
`0.0
`
`PBS Group 2 Group 3 Group 4 Group 5 Group 6 Group 8 Group 9 Group 10 Group 11 Group 12 Group 13 Group 14
`Group 7
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 9 of 77 PageID #: 386
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`FIG . 2
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`U.S. Patent
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`Oct. 12. 2021
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`Sheet 4 of 24
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`3.0
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`2.5
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`2.04
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`
`
`Liver ApoB : GAPD MRNA Ratio 1.5
`
`
`
`
`
`-47 % vs PBS Control
`
`-77 % VS PBS Control
`
`PBS
`
`2:30 SNALP 5x1 mg / kg 1:57 SNALP 5x0.1 mg / kg
`
`1.0
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`0.5
`
`0.04
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 10 of 77 PageID #: 387
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`FIG . 3
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`
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`U.S. Patent
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`Oct. 12. 2021
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`Sheet 5 of 24
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`US 11,141,378 B2
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`Activity of SNALP Upon Intravenous Administration in Mice
`group mean I SD ( n = 4 )
`
`2.0
`
`1.5
`
`
`
`Liver ApoB : GAPD mRNA Ratio 1.0
`
`
`
`
`
`0.5
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`0.0+ PBS Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 11 of 77 PageID #: 388
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`FIG . 4
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`U.S. Patent
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`Oct. 12. 2021
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`Sheet 6 of 24
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`US 11,141,378 B2
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`Activity of SNALP Upon Intravenous Administration in Mice
`group mean I SD ( n = 4 )
`
`2.0
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`1.5
`
`Ratio
`mRNA
`ADOB : GAPD
`Liver
`
`1.0
`
`0.5
`
`0.0 PBS Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9 Group 10 Group 11 Group 12 Group 13 Group 14 Group 15
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`Case 1:22-cv-00252-UNA Document 1-6 Filed 02/28/22 Page 12 of 77 PageID #: 389
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`FIG . 5
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`Sheet 7 of 24
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`US 11,141,378 B2
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`Tolerability of IV 1:57 SNALP in Female BALBIC Mice , 1-4 , SD error
`205
`168
`Alanine Aminotransferase
`Aspartate Aminotransferase
`Sorbital Dehydrogenase
`Incr'd SDH , ALT levels indicate hepatocyte damage .
`96
`
`140
`
`120
`
`100
`
`IUL
`
`80
`
`60
`
`40
`
`20
`
`81
`
`46
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`53
`
`50
`
`40
`
`23
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`27
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`69
`
`59
`
`33
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`27
`
`79
`
`62
`
`58
`
`35
`
`0
`
`Syringe PBS
`Syringe PBS
`Syringe Citrate
`Syringe Citrate
`PBS
`Gear PBS 9mg / kg
`Gear PBS
`7m