UNITED STATES PATENT AND TRADEMARK ()Jj'l?ICE
`
`BEFORE THE PATENT TRIAL AND APPEAL BOARD
`
`LUPIN, LTD. and LUPIN PHARMACEUTICALS INC.,
`
`Petitioner
`
`v.
`
`SENJU PHARMACEUTICAL CO., [,TD.
`Patent Owner
`
`Case IPR2015-01097
`Patent 8, 7 54, 131 B2
`
`DECLARATION OF DARYL S. l'AlJLSON, PH.D., M.A., l\.tl.S., M.B.A.
`
`Page 1 of29
`
`SENJU EXHIBIT 2128
`LUPIN v. SENJU
`IPR2015-01097
`
`

`
`I, Daryl S. Paulson, Ph.D., M.A., M.S., M.B.A., under penalty of perjury,
`
`declare as follows:
`
`I.
`
`QUAijJFICATIONS
`
`1.
`
`I submit this declaration at the request of Finnegan, Henderson,
`
`Farabow, Garrett & Dunner, LLP on behalf of Senju Pharmaceutical, Co., Ltd.
`
`("Senju")
`
`in connection with IPR2015~01097 inter partes review ("IPR")
`
`proceeding before the United States Patent and Trademark Office (''PTO") Patent
`
`Trial and Appeal Board ("Board") as an expert in the field of the evaluation of
`
`drug products. My qualifications in these are::ts, as well as other areas, are
`
`established below and by my curriculum vitae, which is EX2129.
`
`2.
`
`I am currently the President and CEO of BioScience Laboratori.es, Inc.
`
`in Bozeman, Montana, which I founded in 1991.
`
`3.
`
`In the field of microbiology, I received a B.S. degree in medical
`
`microbiology from College of Great Falls in 1979 and a M.S. degree in medical
`
`microbiology/biostatistics from University of Montana in 198 J.
`
`I have also
`
`received a ~1.A. degree in psychology from Pacifica Graduate Institute - Santa
`
`Barbra, California, in 1988, a Ph.D. degree in Psychology from Sierra University
`
`in 1989~ and a Ph.D. degree in Human Science from SayBrook Research Institute
`
`and Graduate School. I have also received an M.B.A. degree from University of
`
`Montana in 2002 1 a Ph.D. degree in Asian art history from Warnhorough
`
`Page 2 of29
`
`2
`
`

`
`University, UK, and a Ph.D. degree m Psychology fnm1
`
`the Institute of
`
`Transpcrsonal Psychology in 2008.
`
`4.
`
`J have an extensive experience in the fields of management science,
`
`research and development, clinical trials, biostatistics, and clinical microbiology
`
`and have authored publications and given presentations related to pharmaceutical
`
`drug products. Among my numerous publications, I have authored Topical
`
`Antimicrobial Testing and Evaluation, Taylor & Francis, 2014~ and I have also
`
`edited the Handbook <~{Topical Antimicrobials, Marcel Dekker, 2002.
`
`H. DOCUl\tiENTS AND INFORlVtATlON CONSIDE~RED IN FORMING
`OPINIONS
`
`5.
`
`Under my direction and control, BioScience Laboratories, Inc.
`
`conducted a Preservative Effectiveness Test ("PETn) to evaluate and determine the
`
`antimicrobial preservatives effectiveness of several samples challenged with
`
`resistant microorganisms. In reviewing the PET results, I had available to me the
`
`documents cited herein as well as the publications listed on my curriculum vitae at
`
`EX2129, The data tables for the PET were created using the raw data collected by
`
`the Study Director, Dan M. Dragotoiu, who conducted this comparative study and
`
`who holds a B.S. degree and has over ten years of professional and academic
`
`experience in the area of microbiology.
`
`I reserve the right to testify about
`
`BioScience Laboratories, Inc. test results and scientific expertise.
`
`Page 3 of29
`
`3
`
`

`
`III. STATElVIENT OF OPI!'I~ONS EXPRESSED AND BASES AND
`REASONS 'fHEREFOR
`
`A.
`
`6.
`
`Background
`
`BioScience Laboratories. Inc. received from SSCI samples of Bausch
`
`& Lomb Incorporated's C'B+L's") Prolensa® product for PET. Upon receipt of
`
`these samples, BioScience Laboratories, Inc. stored, handled, and maintained
`
`according to BioScience Laboratories, Inc.'s cunent good manufacturing practice
`
`("OMP") sample handling procedures as spccificaUy outlined in their SOP L-0005.
`
`These samples were evaluated fot preservative efficacy during the months of
`
`November 2015 and January 2016. BioScience Laboratories Study Director, Dan
`
`M. Dragotoiu, was personally present during the preservative efficacy testing of
`
`these samples.
`
`7.
`
`A summary report of the preservative efficacy testing conducted by
`
`BioScience Laboratories, Inc. on these sarnples is attached as Appendix A. The
`
`report describes the analytical methodology to evaluate preservative efficacy of
`
`stressed and unstressed samples of B+L' s Prolensa® product. The preservative
`
`efficacy results are reported in the document for all of the samples tested,
`
`8.
`
`Bi.oScicnce Laboratories,
`
`Inc.
`
`is a Good Laboratory Practice
`
`("GLP'')/GMP facility providing contract product development services to the
`
`pharmaceutical industry. BioScience Laboratories, Inc.'s service offerings include,
`
`but are not limited to analytical testing (e.g,, preservative efficacy), product
`
`Page 4 of29
`
`4
`
`

`
`development, and manufactures label claims. The preservative efficacy testing of
`
`the B+L's Prolensa® product samples was perfonned in BioScience Laboratories,
`
`Inc.'s GLP/GMP In-Vitro laboratory.
`
`9.
`
`The details of the analytical testing that was performed and the
`
`preservative efficacy test results obtained were provided to SSCI for incorporation
`
`in the SSCI report (Appendix A).
`
`B.
`
`Preservative Efficacy Testing of'B+L's Prolcnsa® product
`
`10. As discussed above, samples of B+L's Pro1ensa® product were
`
`analyzed for preservative efficacy. Consistent with
`
`the
`
`'131 patent, the
`
`preservative effectiveness tests were conducted based on the methods described in
`
`the European Phannacopoeia, using the two bacterial species Pseudomonas
`
`aeruginosa (ATCC #9027) and Staphylococcus aureus (ATCC #6538), and the
`
`two fungal species Candida albicans (ATCC #10231) and Aspergillus brasiliensis
`
`(ATCC #16404).
`
`As indicated in the '131 patent, the preservative efiectiveness of the vials were
`
`evaluated at the following times: 24 hours, 7 days, 14 days and 28 days following
`
`inoculation with the two bacterial species, and 14 days, 21 days and 28 days
`
`Page 5 of29
`
`5
`
`

`
`following inoculation with the two fungal species. 1 (EX2267 at 8.) At these time
`
`points, and after further incubation at the appropriate temperature using the
`
`appropriate agar medium for each species, the colonies on the plates were counted.
`
`tEX2267 at 8-9.)
`
`11.
`
`Some aspects of the method used in this study varied slightly from the
`
`European Pharmacopoeia to account for the low volume of availahle test products
`
`that Lupin and lnnoPharma provided. which were tested at t:he same time with the
`
`B+ L' s Prot en sa \li; product. Still, the test methods are consistent with the '131
`
`patent and scientifically valid, and the results obtained are accurate, as further
`
`described below.
`
`12.
`
`For this study, the test products were transferred from the original
`
`containers to sterilized vials to ensure the accuracy of the volurnes and to maintain
`
`sterility of the
`
`test products at
`
`the various
`
`time points. The European
`
`Pharmacopoeia does not require using
`
`the containers of the test products.
`
`Recognizing that there may be times when the test products are not challenged in
`
`their final containers, the European Pharmacopoeia states that "the test consists of
`
`challenging the preparation, wherever possible in its final container, with a
`
`1 The additional data taken at 21 days conforms to the '131 patent (EX1002
`
`at 9:64 to 10:49 and 14:19-26, 30-38, 42-49, 54-60, and 14:64 to 15:4), and further
`
`confirms the validity of the results of the preservative effectiveness testing.
`
`Page 6 of29
`
`6
`
`

`
`prescdbed
`
`inoculum of suitable micro~organisms, storing
`
`the
`
`inoculated
`
`preparation at a prescribed temperature, withdrawing samples from the container at
`
`specified intervals of tin1e and counting the organism in the samples so removed."
`
`(EX2295 at 505, emphasis added.) Indeed, in the previous paragraph, it also states
`
`that the antimicrobial activity "may" be demonstrated by the described test,
`
`making it clear that the exact conditions of the test can be modified, when
`
`appropriate, and still elucidate a formulation's preservative effi.cacy.
`
`(!d.) For
`
`example, the European Pharmacopoeia allows for different chaHenge species to be
`
`used and provides a subjective evaluation for the sporulation obtained in the
`
`growth of cultures. (/d. at 506, left col., ll. l-23.) Therefore, the use of sterilized
`
`vials for the test products was acceptable fbr this study.
`
`C.
`
`Reported Preservative Efficacy Testing Results are Reliable
`
`1.
`
`1\tleasurement at the Time Points Required by the
`Patent were Made
`
`13. As discussed above, the preservative effectiveness of the vials were
`
`evaluated at the following times: 24 hours, 7 days, 14 days and 28 days following
`
`inoculation with the two bacterial species, and 14 days, 2 I days and 28 days
`
`following inoculation with the two fungal species. (EX2267 at 8.) The tests were
`
`conducted consistent with the ' 131 patent specification, which do not require data
`
`at the zero hour time point.
`
`(See, EXI002 at 9:64-10:49.) Nor is the zero hour
`
`time point a required time point for the EP-criteria B standard as specified in Table
`
`Page 7 of29
`
`7
`
`

`
`5.1.3.- 1. (EX2295 at 506.) In any event, we determined the initial inoculum count,
`
`which was provided in the BSL Fina1 Letter Report. (EX2267 at 25-29.) The lack
`
`of data for the zero hour time point, therefore, would not have any effect on the
`
`accuracy of the results obtained for other data points.
`
`2.
`
`Neutralization Validation vVas Properly Conducted
`
`a. Microorganisms Were Properly Added to the
`Test Product
`
`14. Consistent with the method described in the European Pharmacopoeia,
`
`a 1 ml aliquot of test product was added to 9 ml of the diluent (neutralizer) prior to
`
`the addition of the organism. Thereafter,
`
`Moreover, the microbial recoveries
`
`from these neutralized test products were within a factor of 2 of the recoveries
`
`from the controls, consistent with what is set forth in the European Pharmacopoeia.
`
`(EX2295 at 164-165.)
`
`Page 8 of29
`
`8
`
`

`
`b.
`
`Inoculum Count for .4. niger
`
`15. The inoculum counts for some samples with A. niger (also knmvn as
`
`A. bra,viliensis) for the test products used in the neutralization validation test were
`
`slightly outside the range of the CFUs rcconuncndcd by the method described in
`
`the European Pharmacopoeia. ln all instances, however, the counts recovered from
`
`the test product used in the neutralization validation test did not differ by more
`
`than a factor of 2 from the control, as permitted by the European Phannacopoeia.
`
`(EX2295 at 165, Section 4-5-3 and 4-6.) Thus, the slight variation in CFUs from
`
`the data obtained from the test products in the neutralization validation test met the
`
`above criteria set forth in 4-5-2 of the European Pham1acopoeia. Furthermore, the
`
`recovered populations of A. niger from the test products were not more than 0.3
`
`Iog 10 higher than that of the range set forth in the European Pharmacopoeia, which
`
`is an insignificant difference.
`
`c.
`
`Incubation Time for Some Bacterial Samples
`
`16. The incubation period was longer than the time period set forth in the
`
`European Pharmacopoeia. A prolonged incubation does not decrease the number of
`
`colonies as the incubation continues. Moreover, the pl.:ttc counts obtained do not
`
`exceed countable ranges per SOP L-2059. (EX2294.)
`
`Page 9 of29
`
`9
`
`

`
`3.
`
`Preservative Effectiveness Test was Properly
`Conducted
`
`a.
`
`Initial Inoculum Concentrations were Within
`the Range Allowed by the European
`Pharmacopoeia
`
`17. The European Pharmacopoeia sets forth the use of "an inoculum of
`
`105 to 106 micro-organisms per miJiilitre or per gram of the preparation." (EX2295
`
`at 506.)
`
`concentrations for all of the
`
`microorganisms were within the required range of inoculum concentrations, as
`
`specified in the European Pharmacopoeia. (EX2267 at 41 .) In any event, a high
`
`inoculum concentration just challenges the preservative effectiveness of the
`
`formulation to a greater extent. Eradicating that challenge at the data time
`
`intervals recited in the '131 patent further confirms the preservative effectiveness
`
`of the tested formulations.
`
`b.
`
`Incubation Times Were Aligned with the
`Protocol and/or With Validated Procedures
`
`18. The preliminary counts as well as the final counts were recorded on
`
`the Antimicrobial Effectiveness Testing Data Sheets.
`
`(See, e.g., EX2267 at 44.)
`
`which the counts changed from the preliminary to the final counting dates, those
`10
`
`Page 10 of29
`
`In instances in
`
`

`
`additional plate counts were recorded,
`
`which the counts changed from the preliminary to the final counting dates, those
`
`In instances in
`
`4.
`
`Deviations from the Protocol or Standard Operating
`Procedures in the BSI.J Testing Did Not Impact the
`Results
`
`19.
`
`In my opinion, any deviations from
`
`the Protocol or Standard
`
`Operating Procedures would not have compromised the accuracy or validity of the
`
`results obtained.
`
`a.
`
`Temperature of Molten Agar Did Not Affect the
`Results
`
`20. Although the temperatures for molten agar were slightly higher than
`
`the recommended temperature of 45°C according to the European Pharmacopoeia
`
`(EX2295 at 165),
`
`would not have had an adverse effect on any of the microbial species
`
`tested for this study.
`
`Page 11 of29
`
`11
`
`

`
`variation on these two identif1ed days would not be expected to adversely affect the
`
`ut in the end, this very slight temperature
`
`microbial species tested.
`
`b.
`
`There was Adequate Monitoring of Incubation
`Temperature for the Test Period
`
`21. The incubation was documented to be within the required ranges,
`
`despite that the electronic temperature monitoring system was not in operation
`
`from 11119/15 to ll/29/15.
`
`Furthermore, the electronic temperature monitoring system was
`
`back in operation beginning 11/29/15. In any event, the incubators did not have
`
`any technical issues and were functioning during the time period at issue, and the
`
`temperatures would have remained within the required range of 30-35°C.
`
`c.
`
`Use of Different Medium During Neutralization
`Validation Does Not Invalidate the Obtained
`Test Results for Both Fungi
`
`22.
`
`The media used for each test date for the various microorganisms are
`
`documented in the Media/Diluent Trackjng Forms, which confinns compliance
`
`Page 12 of29
`
`12
`
`

`
`with the testing protocoL (EX2267 at 11; EX2275.) The same media was used for
`
`the neutralization validation test and the preservative etTectiveness test for the test
`
`products.
`
`23. With respect to C. albicans and A. niger, Media/Diluent Tracking
`
`Fonns document the media used on 11/30/15 in which the neutralization was
`
`conducted.
`
`states the use of the TSA+ media for the neutralization validation,-
`
`Page 13 of29
`
`13
`
`

`
`immediately apparent to a person of ordinary skill in the art and does not impact
`
`typographic error would have been
`
`the validity of the results obtained.
`
`24.
`
`can be observed by the tester--"or until good sporulation is obtained." Moreover,
`
`which the EP 7.0 at 506 confirms
`
`According
`
`to Table 2.6.12.-1 of
`
`the European
`
`Pharmacopoeia, either SDA media or PDA media could be used for the cultivation
`
`of A. niger. (EX2295 at 164.)
`
`which is consistent with the description m
`
`the European
`
`Pharmacopoeia.
`
`(EX2295 at 164.)
`
`Page 14 of29
`
`14
`
`

`
`IV. COMPENSATION
`
`26.
`
`If called to testify to the facts stated herein, I will be compensated for
`
`my time preparing for and testifying in this matter at rate of $500.00 per hour. No
`
`part of my compensation is contingent upon the outcome of this matter or any issue
`
`in it.
`
`V.
`
`PRIOR EXPERT TESTIMONY
`
`27. During the past four years, I have not testified as an expert in any
`
`cases.
`
`y~)\?~ - - -
`
`Daryl S. Paulson, Ph.D., M.A., M.S.~ M.B.A.
`
`Page 15 of29
`
`15
`
`

`
`APPENDIX A
`APPENDIX A
`
`Page 16 of29
`Page 16 of 29
`
`

`
`~ sst:r
`
`A Division of Albany Molecular Research Inc.
`
`3065 Kent Avenue
`West Lafayette, IN 47906-1076
`Phone: (765) 463-0112
`Fax: (765) 463-4722
`E-mail: info@ssci-inc.com
`Web: www.ssci-inc.com
`
`Stability Evaluation of
`Bromfenac Sodium Drug
`Product Samples for Potency
`and Preservative Efficacy
`
`Project ID: EL20151326
`Report Date: 01/08/2016
`
`Page 17 of29
`
`

`
`TABLE OF CONTENTS
`
`SUMMARY .......................................................................................................................................................... 3
`I.
`II. RESULTS AND DISCUSSION ........................................................................................................................... 3
`III. DATA TABLES ................................................................................................................................................... 5
`Table 1. Summary ofHPLC Data, Sequence 741881 ................................................................................................. 5
`Table 2. Summary ofHPLC Data, Sequence 742199 ................................................................................................. 6
`IV. EXPERIMENTAL ................................................................................................................................................ ?
`A. HPLC Method ..................................................................................................................................................... ?
`V. APPENDIX A: PRESERVATIVE EFFICACY DATA ...................................................................................... 8
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samplesfor Potency and Preservative Efficacy, 01/08/2016
`page 2 of 13
`Page 18 of29
`
`

`
`I. SUMMARY
`
`Bromfenac sodium ophthalmic solution drug products were sourced from Senju. A portion of
`the samples was used for unstressed (as received) analysis, and the remaining samples were
`stressed in an oven for four (4) weeks at 60°C. Samples from both the unstressed and stressed
`conditions were evaluated for potency and preservative efficacy.
`
`Potency was determined by HPLC with UV detection as detailed in Section IV.A. Percent
`recovery (percent initial) was calculated based on the potency after stress conditions relative to
`that of the unstressed sample. See Table 1 for the unstressed (as received) HPLC data and Table
`2 for the stressed sample HPLC data.
`
`Preservative efficacy was evaluated by BioScience Laboratories, Inc. Samples were evaluated
`for preservative efficacy as guided in the EP Preservative Effectiveness Test Method against the
`following organisms: Candida albicans (AATCC# 10231), Aspergillus niger (AATCC# 16404,
`also referred to as Aspergillus brasiliensis), Pseudomonas aeruginosa (AATCC# 9027) and
`Staphylococcus aureus (AATCC# 6538).
`See Section V for the detailed experimental
`information.
`
`Section II contains results for both potency and preservative efficacy.
`
`II. RESULTS AND DISCUSSION
`
`Material was purchased from one (1) lot, which was used for all experiments and time points.
`Average results are summarized below, with the percent recovery calculated as the amount of
`active relative to the unstressed sample.
`
`Lot Number
`
`240031
`
`Unstressed
`Concentration
`(mg/ml)
`Average= 0.7457
`High= 0.7466
`Low= 0.7451
`
`Stressed
`Concentration
`(mg/mL)
`Average= 0.7445
`High= 0.7473
`Low= 0.7418
`
`%Recovery
`
`Average= 99.8
`High= 100.2
`Low= 99.5
`
`Stressed and unstressed samples were evaluated for preservative efficacy, with the following
`results.
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01/08/2016
`page 3 of 13
`Page 19 of29
`
`

`
`Organism
`
`Condition
`
`S. aureus
`
`P.
`aeruginosa
`c.
`albicans
`
`A. niger
`
`Unstressed
`
`Stressed
`
`Unstressed
`
`Stressed
`
`Unstressed
`
`Stressed
`
`Unstressed
`
`Stressed
`
`Inoculum
`Count
`
`1.97667 X 106
`
`1.7070 X 106
`
`3.3953 X 105
`
`8.8837 X 105
`
`21
`
`---
`
`Cell Count (CFU/ml)
`Days After Inoculation
`2,8
`1
`7
`14
`<1.00 X 101 <1.00 X 101 <1.00 X 101
`<1.00 X 101
`<1.00 X 101 <1.00 X 101 <1.00 X 101
`<1.00 X 101
`<1.00 X 101 <1.00 X 101 <1.00 X 101
`<1.00 X 101
`<1.00 X 101 <1.00 X 101 <1.00 X 101
`<1.00 X 101
`---
`---
`< 1.00 X 101 < 1.00 X 101 <1.00 X 101
`< 1.00 X 102 < 1.00 X 102 <1.00 X 102
`---
`<1.00 X 101 <1.00 X 101 <1.00 X 101
`---
`<1.00 X 101 <1.00 X 101 <1.00 X 101
`---
`
`---
`
`---
`
`---
`---
`---
`---
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01/08/2016
`page 4 of 13
`Page 20 of29
`
`

`
`III. DATA TABLES
`
`Table 1. Summary of HPLC Data, Sequence 741881
`
`lnj#
`
`1
`2
`3
`4
`5
`6
`7
`8
`9
`16
`17
`18
`19
`20
`
`Sample
`Description
`Blank
`Blank
`STDA
`STDA
`STDA
`STDA
`STDA
`STD B
`Blank
`STDA
`240031
`240031
`240031
`STDA
`
`LIMS
`
`LC Filename
`
`408677
`408677
`408705
`408705
`408705
`408705
`408705
`408706
`408677
`408705
`403486
`403489
`403490
`408705
`
`741882
`741883
`741884
`741885
`741886
`741887
`741888
`741889
`741890
`741897
`741898
`741899
`741900
`741901
`
`RT
`---
`---
`8.667
`8.668
`8.666
`8.666
`8.666
`8.665
`---
`8.666
`8.646
`8.648
`8.647
`8.663
`
`Area(mAU*s)
`
`---
`---
`4012.66
`4015.13
`4016.96
`4016.04
`4018.79
`4038.45
`---
`4019.73
`4556.16
`4548.27
`4547.03
`4016.40
`
`SSCI Report: Stability Evaluation of Compound 5 78 Drug Product Samples for Potency and Preservative Efficacy, 01 /08/2016
`page 5 of 13
`Page 21 of29
`
`

`
`Table 2. Summary of HPLC Data, Sequence 742199
`
`Sample
`Description
`Blank
`Blank
`STDA
`STDA
`STDA
`STDA
`STDA
`STD B
`Blank
`STDA
`STDA
`STDA
`STDA
`, - STDA
`
`I
`
`--
`
`STDA
`STDA
`STDA
`240031
`240031
`240031
`240031
`240031
`240031
`STDA
`240031
`240031
`240031
`240031
`STDA
`
`LIMS
`
`LC Filename
`
`408677
`408677
`408705
`408705
`/ 408705
`408705
`408705
`408706
`408677
`408705
`408705
`408705
`408705
`408705
`408705
`408705
`408705
`403478
`403479
`403480
`403481
`403482
`403483
`408705
`403484
`403485
`403487
`403488
`408705
`
`742200
`742201
`742202
`742203
`742204
`742205
`742206
`742207
`742208
`742215
`742222
`742229
`742235
`742242
`742249
`742256
`742262
`742263
`742264
`742265
`742266
`742267
`742268
`742269
`742272
`----
`742273
`742274
`742275
`742276
`
`RT
`---
`---
`8.669
`8.670
`8.670
`8.670
`8.669
`8.669
`---
`8.673
`8.676
`8.677
`8.681
`8.682
`8.684
`8.690
`8.691
`8.670
`8.670
`8.670
`8.673
`8.671
`8.672
`8.694
`8.676
`8.676
`8.678
`8.680
`8.701
`
`Area(mAU*s)
`
`---
`---
`4011.82
`4013.03
`4010.89
`4010.87
`4015.36
`4055.66
`---
`4015.87
`4019.08
`4018.56
`4017.59
`4019.84
`4017.62
`4017.27
`4023.48
`4537.86
`4545.41
`4541.65
`4522.94
`4555.21
`4556.05
`4024.68
`4541.95
`4541.17
`4524.65
`4523.19
`4024.02
`
`SSCJ Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01/08/2016
`page 6 of 13
`Page 22 of29
`
`

`
`IV. EXPERIMENTAL
`
`A. HPLC Method
`
`HPLC analyses were performed using an Agilent 1100 series liquid chromatograph equipped
`with a diode array detector, degasser, quaternary pump, and autosampler. The chromatographic
`column was a Shiseido Capcell Pak C18, 2.1 x 100.0 mm column with 5.0 t..tm packing. The
`column temperature was set to 25°C, and the detector wavelength was 266 nm. The injection
`volume was 5.0 t..tL. The mobile phase was prepared by dissolving 7.9231 g of ammonium
`dihydrogen phosphate into 3000 mL of water, adding phosphoric acid to adjust the pH to 7.30,
`and then mixing in 1000 mL of acetonitrile. The flow rate used was 1.5 mL/minute, with a run
`time of 13 minutes per injection. Method performance was monitored for the following criteria,
`with the listed range of results.
`
`Criterion
`Precision (n=5 %RSD)
`Global %RSD
`Tailing Factor (1st standard injection)
`Plates (1st standard injection)
`Percent Agreement
`
`Results
`0.05%- 0.06%
`0.06% 0.11%
`1.8
`5251-5297
`100.6%- 101.1%
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01/08/2016
`page 7 of 13
`Page 23 of29
`
`

`
`V. APPENDIX A: PRESERVATIVE EFFICACY DATA
`
`SSCI Report: Stability Evaluation a_{ Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01108/2016
`page 8 of 13
`Page 24 of29
`
`

`
`#151142-203.01 Letter Final Rcpm1
`P<~gl! 25 of 65
`
`Protocol #151142-203
`January 07,2016
`
`TABLE 7
`Test Product #3: Prolcnsa'"' bromlcnac ophthalmic solution 0.07%.
`Bromfenac Sodium (unstressed)
`Lot Numbers 240031
`
`I.\0.~.\..P.ro<l.~£! #4,: ProlensaTM brornfenac ophthalmic solution 0.07% (w/v)
`Bromfenac Sodium (4 weeks@ 60"C))
`Lot Number 240031
`
`Challenge
`Microorganism
`(ATCC#)
`
`Initial
`Population
`(CFU/mL
`of Product)
`
`Day(s)
`Following
`Inoculation
`
`Product
`#
`
`Population
`Recovered
`(CFlJ/mL)
`
`Log10
`Reduction
`
`Aspergll/us brasiliensis
`(ATCC#l6404)
`
`8.6818x!05
`
`14
`
`3
`
`4
`
`3
`
`< l.OOx 10 1
`
`4.9386
`
`< l.OOx 101
`
`4.9386
`
`< l.OOx 101
`
`4.9386
`
`4.9386
`
`Acceptance
`Criteria
`l'llet?
`(Yes /No)O
`
`YES
`
`YES
`
`YES
`
`YES
`
`YES
`
`··-·-·~-
`
`Percent
`Reduction
`
`99.9988%
`
`99.9988%
`
`99.9988%
`
`99.9988%
`
`99.9988%
`
`YES
`
`99.9988%
`
`21
`< 1.00 X ]01
`4
`t---~- -· ·-·· . ·········-·
`................... [--
`< l.OOx 101
`3
`4.9386
`-j----···-
`< l.OOx 10 1
`4.9386
`
`28
`
`t-·
`
`4
`..
`·-
`'
`r
`OEuropean Pharmacopoew 7.0. 5.1.3. EFrACACY OF ANTIMICROBIAL PRESERVATION. 0112011.50103. Acceptance Cntena,
`Table 5.1.3.1, Parenteral preparations, eye preparations. intrauterine prepamtions and intramammary preparations: The criteria for
`fungi is a 2 log 1o reduction following 7 days of exposure to the test product with no recovered Colony Forming Units (CFU) recovered
`after 28 days of exposure to the product.
`
`Protocoll/151142-203
`Page 6 of 15
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01/08/2016
`page 9 of 13
`Page 25 of29
`
`

`
`#151142·203.01 Letter Final R._..pot1
`P<1g~ 26 of 65
`
`Protocol!! 151 142-203
`January 07,2016
`
`TABLES
`If§!..Pm<:lt.!<::.LIIJ: Prolensa"'' bromfenac ophthalmic solution 0.07%.
`Bromfcnac Sodium (unstressed)
`Lot Number 240031
`
`Test Product #4: Pro!cnsarM bromfenac ophthalmic solution 0.07% (w/v)
`Bromfenac Sodium (4 weeks@ 60°C)
`Lot Number 240031
`
`Challenge
`Microorganism
`(ATCC#)
`
`Initial
`Population
`(CFU!mL
`of' Product)
`
`Dny(s)
`Following
`Inoculation
`
`Product
`#
`
`Populntion
`Recovered
`(CFU/mL)
`
`Log10
`Reduction
`
`Percent
`Reduction
`
`Candida albicans
`(ATCC#l0231)
`
`3.3182x 105
`CFU/J.Oml.
`
`14
`
`4
`
`21
`
`< 1.00 x 102
`4.5209
`4
`!-------+---+--------.. ·--· · - - -
`< 1.00 X 10 1
`4.5209
`
`28
`
`4
`
`< 1.00 X 102
`
`4.5209
`
`YES
`
`YES
`
`OEuropean l'hannacopocia 7.0. 5.1.3. EFI'ACACY OF ANTIMICROBlAL PRESERVATION. 01/2011:50103. Acceptance
`Criteria, Table 5.1.3.1, Parenteral preparations. eye preparations, intrauterine preparations and intramammmy preparations:
`The criteria for fungi is a 2 log 10 reduction following 1 days of exposure to the test product with no recovered Colony F01ming
`Units (CFU) recovered atler 28 days of exposure to the product.
`
`Protoco 1 #!51 J 42-203
`Page 7 of 15
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative £Yficacy, 01108/2016
`page 10 of 13
`Page 26 of29
`
`

`
`#151142~203.01 Letter Final Rt!pmt
`P<lge 27 of 65
`
`Protocol #151142-203
`January 07, 2016
`
`. TABLE 9
`Test Product #3: Pro!ensa"1 bromknac ophthalmic solution 0.07%.
`Bromfenac Sodium (unstressed)
`Lot Number 240031
`
`Test Em@ct 11·'1: Pro!ensaTM bromfenac ophthalmic solution 0.07% (w/v)
`Bromfenuc Sodium (4 weeks@ 60°C)
`Lot Number 240031
`
`Challenge
`Microorganism
`(ATCC#)
`
`Initial
`Populalion
`(CFU/mL
`of Product)
`
`Day(s)
`Following
`Inoculation
`
`Product
`#
`
`Population
`Recovered
`(CFUimL)
`
`I
`I Pseudomonas aeruginosa
`i
`
`(ATCC #9017)
`
`I
`
`I I ~
`
`I
`
`7
`1.7070 X !06 ---
`
`14
`
`28
`
`Acceptance
`Criteria
`Met?
`(Yes/No)O
`YES
`YES
`YES
`YES
`YES
`YES
`YES
`YES
`'
`'
`OEmopean Phatmactlpoeta 7.0. 5.1.3. EFFACACY OF ANTIMICROBIAL PRESERVATION. Ol/2011.5010o. Acceptance Cntena,
`Table 5.1.3. l, Parenteral preparations, eye preparations, fntra11terine preparations and intrnmammary preparations: Criteria A fOr
`bacteria is a 3 log 10 reduction following 24 hours of exposure to the test product with no recovered Colony Forming Units (CFU)
`recovered after 28 days of exposure to the product.
`
`-
`
`Log10
`Reduction
`
`5.2322
`··------
`5.2322
`
`5.2322
`
`5.2322
`
`5.2322
`
`5.2322
`
`5.2322
`
`5.2322
`
`Percent
`Reduction
`
`99.9994%
`
`99.9994%
`
`99.9994%
`
`99.9994%
`
`99.9994%
`
`99.9994%
`
`99.9994%
`
`99.9994%
`
`<1.00 X 10 1
`3
`-··-··- ·····-~---·· ....... ·--··-···
`<J.OO X 10 1
`4
`<1.00 x 101
`<J.OO X 10 1
`
`3
`
`4
`
`3
`
`4
`
`3
`
`4
`
`<J.OOx 10 1
`<].00 X J0 1
`<J.OO X 10 1
`<].00 X 10 1
`
`Pro loco l 11151142-203
`Page 8 of 15
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01108/2016
`page II of 13
`Page 27 of29
`
`

`
`#.15l142-203.01 LeUer Final Rep01t
`Page 28 of 65
`
`Protocol 11151142-203
`January 07,2016
`
`TABLEIO
`Test P.t"]ld~cttn: Prolcn,a1
`M bromfcnac ophthalmic solution 0.07%
`Bromfenac Sodium (unstressed)
`Lot Number 240031
`
`:C\l~LE.tQ9..\!'-l.!L4: Prolcnsa 1
`" bromfenac ophthalmic solution 0.07% (w/v)
`Bromfenac Sodium (4 weeks@ 60°C)
`Lot Number 240031
`
`Challenge
`Microorgnnism
`(ATCC#)
`
`Initial
`Population
`(CFIJ/mL
`of Product)
`
`Day(s)
`Following
`Inoculation
`
`Product
`II
`
`--
`Population
`Recovered
`(CFU/mL)
`
`Log10
`Reduction
`
`Acceptance
`Criteria
`Met?
`(Yes/No)O
`
`Percent
`Reduction
`
`99.9995%
`
`99.9995%
`
`99.9995%
`
`99.9995%
`
`99.9995%
`
`99.9995%
`
`99.9995%
`
`99.9995%
`
`Staphylococcus aureus
`(A TCC /16538)
`
`I
`1.9767 X J06
`
`I
`
`1
`<1.00 X 101
`4
`5.2959
`~~··--- ---·--·· -··---··-- ··-- ---·-·----- ... ·····--~ ·······--···---
`<1.00 x 101
`5.2959
`3
`7
`
`<1.00 X 10 1
`
`5.2959
`
`<1.00 x 10 1
`
`5.2959
`
`3
`
`4
`
`14
`
`3
`
`4
`
`<1.00 X 10 1
`
`<1.00 x 101
`
`5.2959
`
`5.2959
`
`YES
`
`YES
`·--..--...
`YES
`
`YES
`
`YES
`
`YES
`
`28
`
`····--······~·- ··~··-·-·
`
`5.2959
`
`5.2959
`
`YES
`
`<I.Oo x to'
`3
`[-·-~-"
`<1.00 x 10 1
`YES
`4
`OEurupean Pharmacopocm 7.0. 5.1.3. EFFACACY OF ANTIMICROBIAL PRJC;SERVATION. 01/2011:50103. Accep!aoce Cntena,
`'
`Tolble 5.1.3.1, Parenteral preparations, eye preparations, intrauterh1e preparations and intrarnammary preparations: Critetia A for
`bacteria is a 3 log 10 rcducl!on following 24 hour' of exposure to the test product with no recovered Colony Forming Units (CFU)
`recovered after 28 days of exposure to the product.
`
`Protocol/1151142-203
`Page 9 of .15
`
`SSCI Report: Stability Evaluation a_( Compound 578 Drug Product Samples for Potency and Preservative Efficacy, 01/08/2016
`page 12 of 13
`Page 28 of29
`
`

`
`#151142·203.01 Letter Finall~c-po11
`P<lg~ 29 of 65
`
`Protocol 11151142-203
`January 07.2016
`
`TABLE 11
`Neutralization Evaluation- Results
`:Lc','!._EL'!\h.!(Uc~: Prolensa'M bromfenac ophthalmic solution 0.07%.
`Bromfcnac Sodium (unstressed)
`Lot Numbers 240031
`
`" I ATCC: 1
`Microbial Recovery
`Results
`Challenge Microorg:nism ~#I l'lcr•traliza.~t-.io_'_' =P=b·"--'"--r-(-A_v_e_ra_g;.•c_{_;_F_IJ=·;_p,=Ja_t_e)-l===P=a=s=s/=F=a=il=O==""'l
`CONTROL
`113.00
`---·--·----------+-------------
`Pass
`TEST
`188.00
`..........
`. ..... , __ .. _ .. __________ --!----------··-
`CONTROL
`30.00
`....... ___ -· .. -----·---------1
`30.00
`
`Aspergillus brasiliensis
`
`16404
`
`Candida albicans
`
`10231
`
`TEST
`CONTROL
`
`....... ----·--------- ............ __ .. __ ,,_,_
`
`Pass
`
`Pseudomonas aeruginosa
`9027
`TEST
`-------------+------4-----
`CONTROL
`Staphylowccus aurws
`............................
`6538
`TEST
`23.50
`OThe counts (Average CJ· U/Platc) observed tor the I ES I sample do not vary-by more than a !actor of
`5 ti·mn those recovered from the respective CONTROL sample; hence, the neutralizing media are considered to be
`effective.
`
`30.50
`
`34.00
`
`24.00
`
`.. ..
`
`Pass
`
`Pass
`
`TABLE 12
`Neutralization Evaluation- Result:;
`Tc>i Product IJ4: Prolcnsa""' bromfcnac ophthalmic solt:tion 0.07% (w/vj
`Bromfenac Sodium (4 weeks@ 60°C)
`Lot Nnmber 240031 .
`
`Microbial Recovery
`(Average CFli/Plate)
`
`Results
`Pass/FaiiO
`
`CONTROL
`TEST
`213.50
`r-------------~·~1-"-·----+------------+------------~--------
`CONTROL
`30.00
`TEST
`CONTROL
`
`Aspergillus brasiliensis
`
`16404
`
`Candida n/bicans
`- ......... ·-----------......
`Pseudomonas aeruginosa
`
`10231
`
`9027
`
`113.00
`
`36.00
`
`Pass
`
`f'ass
`
`TEST
`CONTROL
`TEST
`0 The counts (Average CFU/Piate) observed for .... th~-·frs:r sample do not vary by more than a factor of
`5 from those recovered from the respective CONTROL sample; hence) the neutralizing media arc considered
`to be cffccti vc.
`
`Staphylococcus aurcus
`
`6538
`
`·--~-·-~~
`
`Protocol #151142-203
`Page 10 of 15
`
`SSCI Report: Stability Evaluation of Compound 578 Drug Product Sample