~
`
`CRO SS
`
`L
`
`I NG~ "' L
`
`CROSSUNGUAl. UC
`3736 Fallon Road, Suite 226
`Dublin, CA 94568
`Tel: 206-851-7932
`hr@cllnsual.com
`
`AFFIDAVIT OF TRANSLATION
`
`I, Alan F. Siegrist, of CROSSlINGUAl, llC, hereby declare that:
`1. I am fluent in Japanese and English.
`2. I am an active member of the American Translators Association and a Certified
`Translator of Japanese to English.
`3. The English translation attached to this declaration is an accurate and correct
`translation of the following document, attaclled hereto:
`Sawa Declaratlon_2.18.2016
`I declare that the foregoing Is true and correct to the best of my knowledge.
`
`Executed on February 19, 2016
`
`Alan F. Siegrist, cr
`CROSSlINGUAl, llC
`ATA Member No. 31889
`Certification #63788
`
`\Iorf\'. __ ~
`
`A notary public or other officer completing this cenlflcate verIfies only the IdentIty of the IndIvIdual who signed the
`documents to whIch thIs certIficate are a!tathed, and not the truthf~lne ss, <lewra, , or valid! of that document.
`
`State of California, County of C.orrfrq c.e:~J'(!
`-a 1\
`bef",e~e,~b.eJ~A?~~,\AMj Nat~~ \!Ali t
`o,k&I!,.qCy 1\;11216
`personally appeared (~Ian I. ::, ~fl J
`
`who proved to me on the basis of satisfactory
`evidence to be the person whose name is ubscrlbed to the within instrument and acknowledged to me
`that he executed the same In his authorized capacity. and that by his signature on the Instrument the
`person, or the entity upon behalf of which the person acted, executed the instrument.
`
`I certify under PENALTY OF PERJURY under the laws of the State of California t hat the foregoing
`paragraph Is true and correct.
`
`Signature G~~2:.--.'~~==:::=--- (Seal)
`
`Page I of613
`
`SENJU EXHffiIT 2098
`LUPIN v SENJU
`IPR2015-01l05
`
`

`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`
`BEFORE THE PATENT TRIAL AND APPEAL BOARD
`
`LUPIN LTD. and LUPIN PHARMACEUTICALS INC ..
`
`Petitioner,
`
`v.
`
`SENJU PHARMACEUTICAL CO., LTD.
`
`Patent Owner.
`
`Cases IPR2015-0 1099, IPR2015-01097, IPR2015-01105 & IPR2015-01100
`
`Patents 8,669,290, 8,754,131, 8,871,813 & 8,927,606
`
`DECLARATION OF SHIROU SAWA
`
`Page 2 of613
`
`

`
`I, Shirou Sawa, under penalty of perjury, declare as follows:
`
`I.
`
`INTRODUCTION
`
`1.
`
`2.
`
`I am of legal age and otherwise competent to make this declaration.
`
`I am the first named inventor on U.S. Patenl Nos. 8,669,290
`
`("the '290 patent"), 8,754,131 ("the ' 131 patent"), 8,871,813 ("the '813 patent")
`
`and 8,927,606 ("the '606 patent"). I have been asked to submit a declaration
`
`attesting to how the data provided below, including data disclosed in the
`
`specification of the '290 patent, the' 131 patent, the ' 813 patent and the '606 patent,
`
`were generated.
`
`II.
`
`EDUCATION AND WORK EXPERIENCE
`
`3.
`
`I graduated from the University ofTokushima, Tokushima, Japan in
`
`1988, with a bachelor's degree from the Department of Chemical Engineering, in
`
`the Faculty of Engineering. I received a master's degree in engineering, with
`
`chemical engineering specialization, from the University ofTokushima,
`
`Tokushima, Japan in 1990.
`
`4.
`
`I am currently employed by Senju Pharmaceutical, Co., Ltd. ("Senju")
`
`and have been with Senju since 1990. I have worked at Senju as a researcher in
`
`the ophthalmic formulations area from 1990 to the present.
`
`Ill. DATA, INCLUDING DATA IN THE '290, '131, '813 and '606
`
`PATENTS
`
`Page 3 of 613
`
`2
`
`

`
`5.
`
`I was involved in fonnulating and testing bromfenac sodium (sodium
`
`2-amino-3-( 4-bromobenzoyl)-phenylacetate) fonnulations, including the
`
`fonnulations of the Experimental Examples reported in the '290, ' 131, ' 813
`
`and '606 patents. As such, I have first-hand knowledge of how the fonnulati ons
`
`reported below were made and tested.
`
`6.
`
`The laboratory notebooks in Appendices A, Band C describe the
`
`tested fonnulations and the experimental test results obtained with them.
`
`A.
`
`Stability d.ta, including data from Table 1 of the '290, '131, '813
`.nd '606 patents
`
`7.
`
`I prepared and tested the stability of the bromfenac sodium
`
`fonnulations disclosed in Table I of the '290, '\31 , '813 and '606 patents. The
`
`table below contains bromfenac sodium fonnulations from Table 1 of the patents,
`
`as well as additional formulations with varying amounts oftyloxapol. (Appendix
`
`A.) As the table indicates, ronnulations A-20, A-21 and A-27 correspond to
`
`Comparison Examples I, A-02 and A-03, respectively, from Table I of
`
`the '290, ' 131 , '8 13 and '606 patents.
`
`8.
`
`As reflected in the laboratory notebook of Appendix A, the stability of
`
`these bromfenac sodium fonnulations was tested after adjusting the pH of the
`
`fonnulations to 7. Using these fonnulations, accelerated stability tests were
`
`conducted for various lengths of time and elevated temperatures, including for four
`
`weeks at 60 °e. The percent ofbromfenac sodium remaining was measured using
`
`Page 4 of 613
`
`3
`
`

`
`a High Perfonnance Liquid Chromatography ("HPLC") method under the
`
`following conditions:
`
`• Ultraviolet absorbance spectroscopy at 266 nm;
`
`• Column: Capcelpak column;
`
`• Column temperature: 25 °C;
`
`• Mobile phase: 1.98 g of ammonium dihydrogen phosphate was dissolved
`
`in 750 mL of water, the pH was adjusted to 7.3 by adding phosphoric
`
`acid, and 250 mL of acetonitrile was added;
`
`• Flow rate: Adjusted so that the elution time ofbromfenac sodium
`
`becomes 18 minutes; and
`
`•
`
`Injection volume of sample: 10 !(L
`
`9.
`
`The percent ofbromfenac sodium remaining in each formulation after
`
`four weeks at 60°C is tabulated below. The percent ofbromfenac sodium
`
`remaining was adjusted to take into account the amount of water evaporation from
`
`the formulation .
`
`Page 5 of 613
`
`4
`
`

`
`A-20
`
`0.1 g
`
`1.5 g
`
`Formulation code
`Designated code in
`Comparison
`rable I of
`the '290, '13 1, '8 13 Example 1
`and '606 vatents
`Bromfcnac sodium
`hydrate I
`Boric acid
`Benzalkonium
`chloride
`Polysorbate 80
`Tyloxapol
`Sodium hydroxide
`Disti lled water
`Total amount
`pH
`60 °C - 4 weeks
`
`0.005 g
`
`0. 17 g
`-
`q.s.
`q.s.
`100 mL
`7
`51.27%
`
`A-21
`
`A-27
`
`A-28
`
`A-29
`
`A-02
`
`A-03
`
`N/A
`
`N/A
`
`O. I g
`
`1.5g
`
`0.005 g
`-
`0.15 g
`q.s.
`q.s.
`100mL
`7
`73.81%
`
`0.1 g
`
`1.60
`
`0.005 g
`
`-
`0.02 g
`q.s.
`q.s.
`100 mL
`7
`89.64%
`
`O. I g
`
`1.6 g
`
`0.005 g
`-
`0.05 g
`q.s.
`q.s.
`100 mL
`7
`85.96%
`
`0. 1 g
`
`1.6 g
`
`0.005 g
`
`-
`0. 1 g
`q.s.
`q.s.
`100 mL
`7
`82.01%
`
`B.
`
`Stability data, including data disclosed in Table 2 of
`the '290, '131, '813 and '606 patents
`
`10.
`
`[prepared and tested the stability of the following bromfcnac sodium
`
`formu lations, including those disclosed in rable 2 of the '290, , 131 , '813 and '606
`
`patents. As indicated in the following table, formulations A-OI , A-02 and A-03
`
`correspond to formulations A-04, A-OS and A-06, respectively, from Table 2 of
`
`Ute '290, ' 131 , '813 and '606 patents. Stability tests on these formulations were
`
`carried out for various lengths of time and elevated temperatures, including four
`
`weeks and 60 Qe, adjusted to a pH of about 8. t 5, as indicated. The percent of
`
`bromfenac sodium remaining in each formulation after four weeks at 60 °e was
`
`I Tn the appended laboratory notebooks, I used the shorthand "bromfenac sodium"
`to refer to "bromfenac sodium hydrate."
`
`Page 6 of613
`
`5
`
`

`
`measured using the HPLC method described above, which was adjusted to take
`
`into account the amount of water evaporation. The data are tabulated below.
`
`(Appendix C.)
`
`Formulation code
`Designated code in Table 2 of
`the '290, ' 13 1, '813 and '606
`I patents
`Bromfenac sodium hydrate
`Boric acid
`Borax
`Benzalkonium chloride
`Tyloxapol
`Polyviny Ipyrrolidone
`Disodium edetate
`Sodium hydroxide
`Distilled water
`Total amount
`jpH
`60°C - 4 weeks
`
`A-OI
`
`A-04
`
`0.1 g
`1.1.
`1.1 •
`0.005 g
`0.02 g
`2.0 g
`0.02 g
`a.s
`a.s.
`100 mL
`8.15
`92.57%
`
`A-02
`
`A-OS
`
`0.1 g
`1.1.
`1.1 •
`0.005 g
`0.05 g
`2.0 g
`0.02 g
`a.s
`a.s.
`100 mL
`8.15
`90.93%
`
`.
`A-03
`
`A-06
`
`0.1 g
`1.1.
`I.1g
`0.005 g
`0.D3 g
`2.0.
`0.02.
`a.s.
`Q.S.
`100mL
`8.15
`91.97%
`
`C.
`
`Stability data for bromfenac sodium formulations containing
`polysorbate 80 and bromfenac sodium formulations containing
`tyloxapol
`
`11.
`
`I prepared and tested the following bromfenac sodium formulations,
`
`containing the components and amounts as indicated below. Stability tests on
`
`these formulations, adjusted to a pH of about 8.2 to 8.3, were carried out for
`
`various lengths of time and elevated temperatures, including for four weeks and
`
`60 °C, as indicated. TI.1e percent ofbromfenac sodium remaining in each
`
`formulation atter four weeks at 60 °C was measured using the HPLC method
`
`Page 7 of 613
`
`6
`
`

`
`described above, which was adjusted to take into account the amount of water
`
`evaporation. The data are tabulated below. (Appendix C.)
`
`FOimulation code
`
`Bromfenac sodium hydrate
`Boric acid
`Borax
`Benzalkonium chloride
`Polysorbate 80
`Tyloxapol
`Polyviny Ipyrrolidone
`Disodium edetate
`Sodium sulfite
`Sodium hydroxide
`Distilled water
`Total amount
`'DH
`60 °C - 4 weeks
`
`BF (PE)'
`(Broil ti ck)
`0. 1 g
`I.1g
`1.1 .
`0.005.
`0.15 g
`-
`2.0 g
`0.02.
`0.2.
`q.S
`q. S.
`100 mL
`8.20
`9 1.45%
`
`A-O I (PE)
`
`A-03 (PE)
`
`0. 1 g
`I.lg
`1.1 .
`0.005.
`-
`0.G2 g
`2.0 g
`0.02.
`-
`q.S
`q. S.
`100 mL
`8.20
`93.61 %
`
`0.1 g
`I.lg
`I.1g
`0.005 g
`
`-
`
`0.03 g
`2.0 g
`0.02.
`-
`q.S.
`q.S.
`100mL
`8.27
`95 .07%
`
`D.
`
`Preservative efficacy of bromfenac sodium formulations
`
`12.
`
`I was involved with testing the preservative efficacy ofbromfenac
`
`sodium formulations as part of projects P2002B 116 and P2002B 131 at Senju. I
`
`fonnulated a bromfenae sodium [onnulation containing polysorbate 80 (identical
`
`to Bronuek) and bromfcnac sodium fonnulations containing tyloxapol as shown in
`
`the following table. (Appendices B & C.) Formulations A-O I and A-02 in this
`
`table arc the bromfcnac sodium fonnulations containing ty loxapol and correspond
`
`2 "PE" in these designations signifies that the stability test was conducted using
`polyethylene containers.
`
`Page 8 of613
`
`7
`
`

`
`to fonnulations A-04 and A-OS, respectively, from Tables 2, 3- 1 and 3-2 of
`
`the '290, , 13 1, ' 8 13 and '606 patents:
`
`Formulation Code
`Designated code in Tables 2, 3-
`1 and 3-2 of
`the '290, ' 13 1, ' 8 13 and '606
`[patents
`Bromfenac sodium hydrate
`Boric acid
`Borax
`Benzalkonium chloride
`Polysorbate 80
`Tvloxaool
`Polvvinv lovrro l idone
`Disodium edetate
`Sodium sulfite
`Sodium hydroxide
`Distilled water
`Total Amount
`pH
`
`Bronuck
`
`A-OI
`
`A-02
`
`N/A
`
`0. 1 g
`l.l g
`l.lg
`0.005 g
`0.15 g
`-
`2.0 0
`0.02 g
`0.2 g
`q.s.
`q.s.
`100 mL'
`8.3
`
`A-04&
`Table 3-1
`
`A-05&
`Table 3-2
`
`0.1 g
`l.l g
`l.I g
`0.005 g
`-
`0.02 g
`2.0 g
`0.02 g
`-
`q.s
`q.s.
`100 mL
`8. 19
`
`0.1 g
`l.I g
`l.I g
`0.005 g
`-
`0.05 g
`2.0 g
`0.02 g
`-
`q.s
`g.s.
`100 mL
`8.20
`
`13. The preservative efficacy of the bromfenac sodium formulatio n
`
`containing polysorbate 80 (identical to Bronuck) was tested as part of project
`
`P2002B 11 6 at Senju. (Appendix B.) The preservative efficacy of the two
`
`bromfenac sodium fo rmulations containing ty loxapol (formulations A-04 and A-OS
`
`from Tables 2, 3-1 and 3-2 of the '290, , 13 1, , 813 and ' 606 patents) was tested as
`
`part of project P2002B 13 1 at Senju. (A ppendix C.) The preservative efficacy tests
`
`3 In Appendix B. the concentration of each component is disclosed as amount of
`component per 1 mL of the formulation. The value is converted to the amount o f
`each component per 100 mL o f the formulation.
`
`Page 9 of 613
`
`8
`
`

`
`were conducted at Senju using the European Pharmacopoeia (UE?" ) standards, and
`
`I reviewed the results obtained from this testing. (Appendices B & C.)
`
`14. According to the EP standards, the tested formulations were
`
`distributed in sterilized test tubes with stoppers. For inoculation of the organisms,
`
`suspensions containing 108 colony forming units ("CFU")/mL were prepared for
`
`each bacterial species, and suspensions containing 10' CFU/mL were prepared for
`
`each fungal species. Each test formulati on was inoculated in a separate test tube
`
`with a final concentration of 10' CFU/mL of bacteria and 10' CFUlmL of fungi.
`
`The test tubes were stored at 20 to 25 °C after inoculation. Samples were taken
`
`from each test tube after 6 hours, 24 hours, 1 week, 2 weeks, 3 weeks and 4 weeks.
`
`From each sample, 0.5 mL of the sample was diluted with 4.5 mL of sterilized
`
`isotonic sodium chloride solution. Ten-fold dilution was perfonned 1 to 3 times,
`
`and 1 mL of the diluted solution was placed onto a plate, and 15 to 20 mL of a
`
`culture medium was added to each plate. With respect to bacteria cultures,
`
`soybean-casein digest agar medium (SCD agar medium) containing inactivators
`
`(0.1 % lecithin, 0.7% polysorbate 80) was used. With respect to fungi cultures,
`
`sabouraud's glucose medium containing inactivators (0. 1 % lecithin, 0.7%
`
`polysorbate 80) was used. Cultures were kept under the following conditions, and
`
`the number of microorganisms was counted.
`
`Page 10 of 613
`
`9
`
`

`
`Microoro anisl1l
`Staphylococcus aureus A Tee
`6538
`Escherichia coli A Tee 8739
`Pseudomonas aeruginosa ATCC
`9027
`Candida albicans A Tee 1023 1
`AsoerJdllus niger A Tee 16404
`
`Culture Condition
`
`30- 35 'e
`
`20- 25 'e
`
`Bacteria
`
`Fungi
`
`t 5. The results obtained from these preservative efticacy tests are as
`
`follows (Appendices B & C):
`
`Cell count (CFU/mL)
`14 days 2 1 days 28 days
`Inoeul 6 hours 24 hours
`7 days
`afier
`after
`after
`after
`after
`after
`um
`inoculati
`inoculati
`inoculati
`inocu lati inoculati inoculati
`Count
`on
`on
`on
`on
`on
`on
`4.3 x
`4.0 x 10' 3.1 x 10' 7.0 x 10'
`10'
`6.9 x
`10'
`1.1 x
`10'
`3.2 x
`10'
`1.1 x
`10'
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`2.5 x 10'
`
`0
`
`-
`-
`
`0
`
`0
`
`-
`-
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`4.7 x 10'
`
`6.0 x 10' 3.0 X 10'
`
`Bronuck
`
`S. aureus
`
`E. coli
`
`P.
`aeruf!inosa
`
`C. albicans
`
`A. niger
`
`Page 11 of 613
`
`10
`
`

`
`Cell CO llil l (CFU/mL)
`6 hours 24 hours 7 days
`14 days 2 1 days 28 days
`aHer
`after
`after
`a:fter
`a fief
`after
`inocu lali inoculati inoculati inoculati
`inoculati
`inoculati
`on
`011
`011
`0 11
`011
`011
`2.1 x 3.0 x 10'
`10'
`6.5 x
`10'
`5.8 x
`10'
`3.2 x
`10'
`1.8 x
`10'
`
`0
`
`0
`
`0
`
`-
`
`-
`
`0
`
`0
`
`-
`
`-
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`Tnoeultt
`III
`Count
`
`Table 3-1
`A-OJ (A-
`04 in
`spccificati
`0 11)
`
`S. aureus
`
`E. coli
`
`P.
`aeruginosa
`
`C. a/bicans
`
`A. niger
`
`Table 3-2
`A-02
`(A-04 in
`specificati
`on)
`
`S. aureus
`
`E. coli
`
`P.
`aeruKinosa
`
`C. albicans
`
`A. niger
`
`Ccil COUllt (CFU/mL)
`6 hours 24 hours
`14 days 2 1 days 28 days
`7 days
`Inoeul
`after
`after
`aller
`aller
`after
`after
`um
`inoculati
`inoculat i inoculati
`inoculati
`inoculati
`inoculati
`Count
`on
`on
`on
`on
`0 11
`011
`2.1 x 1.7 x 10' 2.0x 10'
`10'
`6.5 x
`10'
`5.8 x
`10'
`3.2 x
`10'
`1.8x
`10'
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`-
`-
`
`0
`
`0
`
`-
`-
`
`16.
`
`The EP-A and EP-B standards used to compare the obtai ned results, at
`
`the time of testing, are the following:
`
`II
`
`Page 12 of 613
`
`

`
`A standard
`
`B standard
`
`Viable cell counts of
`fungi (c. albicans & A.
`niger) 7 days after
`inoculation decreases to
`not more than 11100, and
`the cell count levels off or
`decreases thereafter.
`
`European Pharmacopoeia Standards
`Viable cell counts of
`bacteria (S aureus, E. coli
`& P. aeruginosa) 6 hours,
`24 hours and 28 days after
`inoculation decrease to
`not more than 11100, not
`more than 111000, and
`undetectable resDectivelv.
`Viable cell counts of
`bacteria (8. aureus, E. coli Viable cell counts of
`fungi (c. albicans & A.
`& P. aeruginosa) 24
`hours and 7 days after
`niger) 14 days after
`inoculation decrease to
`inoculation decreases to
`not more than I I I 0 and
`not more than 1/10, and
`not more than 1/ 1000, and the cell count levels off or
`the cell count levels off or decreases thereafter.
`decreases thereafter,
`
`17. Based on the EP-A and EP-B standards, the bromfenac sodium
`
`fonnulation containing polysorbate 80 did not pass either EP standard, The
`
`bromfenac sodium formulation containing tyloxapol and designated A-O t
`
`(corresponding to A-04 from the '290, , 13 1, '8 13 and '606 patents) passed both
`
`standards. The bromfenac sodium formulation containing tyloxapol and
`
`designated A-02 (corresponding to A-OS from the '290, , 13 1, '813 and '606
`
`patents) passed the EP-B standard but not the EP-A standard.
`
`E.
`
`The data in the specification and data disclosed in this declaration
`
`18. Based on my involvement with the formulations of Tables I, 2 and 3
`
`of the '290, ' 13 1, ' 813 and '606 patents, I have personal knowledge about how the
`
`Page 13 of 613
`
`12
`
`

`
`test data disclosed in the ' 290, , 131 , ' 81 3 and '606 patents were generated. Based
`
`on my involvement with the remaining formulations and test data (beyond those
`
`disclosed in Tables 1, 2 and 3 of the '290,. ' 131, '813 and ' 606 patents) provided in
`
`this declaration, including in Appendices A, Band C, I have personal knowledge
`
`about how such information was generated.
`
`19.
`
`I hereby declare under penalty of perjury under the laws of the United
`
`States of America that the foregoing is true and correct, and that all statements
`
`made of my own knowledge are true and that all statements made on information
`
`and belief are believed to be true.
`
`Date: 021181'16
`
`Shiro" Sawa [signature]
`Shirou Sawa
`
`Page 14 of 613
`
`I )
`
`

`
`LUPIN LTD. &11
`LUPIN PHARMACEUTICALS INC.
`ili:ilt).,
`
`v .
`
`• i4 IPIUOI5-01099, IPR2015-01097, IPR201 5-01105 & IPR2015-01100
`~\'il'r 8,669,290, 8,754,131 , 8,871,813 & 8,927,606
`
`Page 15 of 613
`
`

`
`fA, ililfj~I;!: , ia'!!HI'-c])$ljl\lto) r I:::, ~J r O)c;t:; L) i1i: i!i' ,t=L..a:"t •
`'*1<
`
`I.
`
`I.
`
`fA I;!; , lilt).. L.. "(;BL) , it!!I:::t:::O)]r~ .~iflilt"t ~~~:I:J I:::~ It~:::c
`
`1;!:iV>L)a:1t Iv.
`
`2.
`fAl;!:, *OOIH'fii..,. 8,669,290 (r290 iH\li'f J), 8,754,131(r 131 ~
`m'f J), 8,871,813(r81 3 ""lli''!' J)& v: 8,927,606(r 606 -li}1\l·,If J)O)1lu~aJl'ill'-C;
`"t. fkl;!: , 290 ..,.,\lilf, 131 ""11i!'f, 813 -li}l1i,'!' & v: 606 ..,.,\lilfO) aJlta.I:::!~
`I£~;tt. n '~T-9~a-t.'r8CO)T-9tJ{c-O)J:?I:::L.. "( !t1ilt~;tt.t=tJ'~liEg"t
`~ i1i: a -!'H~tf:l"t ~ J:? I::: ~*i'l~;tt. a: L.. f=.
`ftl!Ut V:.1f!
`
`11.
`
`3.
`
`fA I;!; , 8 *OOli,Ol, !/j\I:::pfrll"t ~Ii§j;;!!f'~ 1988l1'l:::I!!f'lIBit!!f'
`
`I !!f'f4 ~ 2?i. iii! L.. a: L.. f=. fA I;!:, I!!f'~ ±..,.1t ~ I !!f'.I'PJt ~ 8 * 00 Ii .. !Iil! :::pfr II
`
`"t ~li~j;;!!f'tJ'o:, 1990 lI'l:::JIl(~L..a:L..f=.
`
`4.
`
`fA I;!; , !Jlll, 'fln'l~,*'i.t:jl;tW'f~JlI:::JiIll~;tt. "(;BL) , 1990
`
`lI'tJ,o:,'Hfl :::iJjffiL.. n 'a:"t. fA I;!: , 'f~I:::"( 1990 lI'tJ'o:,~lla:-C;Ul!HIll~
`tlllI:::~nMJf~'ill'cL.. "(w. 'n 'a:"t 0
`III. 290 -li}, 131 -li}, 813 -Ii} & v: 606 ~*I'R'fO)T-1r~~t.'T-1r
`
`2
`
`Page 16 of613
`
`

`
`5.
`
`fkl;l:. 290 ~. IJ 1 ~. 81 3 ~ lil.Lf 606 ~'I\¥~'fI::~&'a~nt=71J'£":>
`
`IT?T~I)? '£"(2-7"$./ -3-(4-71J'I:,-{;")' l' )~ ) :>I-=-)~fFMT~I)? '£")0)m:i1i1j
`
`0). ~nO)lI!~j;01I~t;~t=~Ii'l1tlil.Lfiit~~I::ll!ItN*I.J= o f:O)t=/IJ. fW:.
`
`r!~I::~'a~ntd~~Ii'ltJlcO)&:'5I::L -C1'F';;f1.. ii(~~nt=tJ'O)iiHjHY.Jt';:~Diat
`
`~I~-:>n '*1'0
`
`6. m:;1>titn A. B lil.Lf C O)~~/-~tJ{. i>t~~nt=~Ii'Ilil.Lff:nl::
`
`&:-:>-cj~';nt=a;t~*"!I!HcitL -n '*1' 0
`
`A_ 290~. 13I~. 813 -tt&Lf 606 -&lII"i'FO)lI! I iil<O)'f-;~at;~lEtt
`
`'f-;
`
`7.
`
`fkl;l:. 290 -&. 131 -&. 813 -& lil.Lf 606 -&~;!;g'fO)~ 1 ~1::1lIlff-~n
`
`t= 71J'£":> IT?T~I)? '£"m:1Ii~O)*{r,jL. ~lEtt~iit~L*Lf=o r~cO)i<I;l:.
`
`~;!;&'f0):;g 1 ~0):11J.£,. :>IT?T~I)? '£"~Ii'lt=lt1:t,;:<. 11!!1::,w,t,;:-:>t=Ii:O)'T
`
`o'l'--It;f<-)~~at;m:1Ii'I~~ A, 1:, '*1' (m:;f>t-=iIA A) 0
`
`iil<tJ(ff-T &:'51::. m:11il1
`
`A-20. A-21 lil.Lf A-27 I;l: 290 ~. 131 -&. 813 -& lil.Lf 606 ~'I\¥a'FO):;g, ~O) It
`
`*:i101J I. A-02lil.Lf A-03 1::f:n-fnMr.I;L n '*T ,
`
`8.
`
`m:;1>t1l=n A O);Ii;.'IV-H::&~~n n '-<>&:'51::. ;:n';71J'£":>I
`
`T?T~IJ?'£"m:11i'10)~lEl11;l:~1iIj~ pH71::tlllf/!'Lt=j~iit!~~n*Lf=, ;:n
`.; ~ /i'l ~ ~, '. 60 'c 1: l1!l;!ll r., ~ B!i ~ n 1='1 .. 0) ~ t; t; . i* I.< f,j: ~ r.,. ;l[ J3f.t. ¥I-r
`
`3
`
`Page 17 of 613
`
`

`
`T:(1), ho;!~:iE11liU~.,~1il!iLto o 1~:{f:101. 7 I -r?-r f-I) '7 1.(1)1 (--I:!:..-f(cid:173)
`
`Ii, 1'~(1)~{<t-1' I::, ~11ligiill*?o "f-?"'771'-(fHPLCJ);'1&IIlL '"(i~II:iE~
`
`nil:Ltoo
`
`• ~~P.&:It:'J'j't:ltiJ'otT:iJ!l:& 266 nm
`
`•
`
`iJ'71. : Capcclpak iJ'71.
`
`• fUM!!: 1):"-~2*JIl7:"-t=-'71. 1.98 g" * 750 mL l::itML, pH
`
`"') :..-M.,hO;t "( 7.3 1::i1ll!'lLtolf , 250 mL (1)7-t! f-::.f-I) 1~.,IJil :Uo
`
`• iM;!: :101.7I-r?-rf-')'71.l1) i3tfj~r.'tJI18 :$l'l:tJ:'QJ:?I::iJIJ~
`
`•
`
`t>t*4(1) ;lA :&:: 10 ~L
`
`9. * :101.7I-r?-rf-')'71.!1l\ljl1) 60°CT:I1!l;!!lMlfl::1~?n '.Q:1
`
`ok 7I-r?-rf-
`
`'
`
`) '71.(1)1 (--I:!:..-f-., 1'Ic~I::fc!L *Ltoo ;::(1)tiff:11J 1. 7I
`
`-r?-rf-') '7 1.(1)1 (--I:!:"-Hi*:$l'",,~ • .,liliiELto t(1)T:iI»'JiI:"t 0
`
`:
`
`4
`
`Page 18 of613
`
`

`
`I:!~IJ::J-i"
`290~, 131~, 813
`~&v: 606 ~~;¥a'f0)
`111 I J\< I ::J\< iC O)::J -
`i"
`7DL. 7:J:T?T~I)
`'/ L.lKlOlo/J I
`*'/1iJl
`!N.1t~:,"1f )~::J_ '/
`L.
`;f<1)~)~~-~ 80
`7D"'<t;f<-)~
`lKM1tT~I),/L.
`iiHlllK
`:i:li
`I pH
`60 °C - 4 i!ll
`
`A-20
`
`A-21
`
`A-27
`
`lt~jjlJ I
`
`A-02
`
`A-03
`
`0.1 g
`
`1.5 g
`
`0.005 g
`
`0.17 g
`-
`i.U:
`iMit
`100 mL
`7
`51.27%
`
`0.1 g
`
`1.5 g
`
`0.005 g
`-
`0.15 g
`iMm
`iMli:
`100 mL
`7
`73.81%
`
`0.1 g
`
`1.6 g
`
`0.005 g
`
`-
`0.02g
`iMiii:
`JM:i:
`100 mL
`7
`89.64%
`
`A-28
`
`..
`
`0.1 g
`
`1.6 g
`
`0.005 g
`-
`0.05 g
`iMii
`iMii
`100mL
`7
`85.96%
`
`A-29
`
`..
`
`0.1 g
`
`1.6 g
`
`0.005 g
`-
`0.1 g
`iMiI!:
`iMll
`IOOmL
`7
`82.01%
`
`813 ~&v: 606 ~';¥~'f0)1II 2J\<0) A-04, A-05:&V: A-061::i-:tLi':tL~r.l;L L
`
`1 i;!l1i2':tL LL '.Q~~V-H::I;I:, fJ.l;l:f7DL. 7:J:T?T~I) '/ L.lKlO'!o/JJo)~1:
`~ f 7DL. 7:J:T?T~I)'/L.JI:'iOSl\~w!~m~ ,too
`5
`
`Page 19 of 613
`
`

`
`r
`!ll!J\lJ :oJ -
`290-1}, 131-1}, 813"'&0'
`606 -I}¥f~ 0)1I! 2 "'1::31<12
`O):J-t:
`7'pA7IT"TI- ~?A
`7J<,fnMl
`7];?1Ii1
`*?iJ;
`IlHt"'" / -if I" :oJ -=- ? A
`'TP""<t* I"
`ij~!J t'"':='/L-- t o t1 !J J.: /
`:r. ,7 H !lj- I- ~ ? A
`IKlIi1it T I- ~ ? A
`#i~A<
`"i'!:-!I(
`pH
`60 ' C - 4 i!!l
`
`A-O'
`
`A-04
`
`0.1 g
`
`Ll g
`LIg
`0,005 g
`0,02 g
`2,0 g
`0,02 g
`in:
`illiiK
`100 mL
`8,15
`92.57%
`
`A-02
`
`A-OS
`
`0,1 g
`
`Llg
`LIg
`0,005 g
`0,05 g
`2,0 g
`0.02 g
`;Mit
`jfj'lll:
`100 mL
`8.15
`90.93%
`
`A-03
`
`A-06
`
`0.1 g
`
`Ll g
`LIg
`0,005 g
`0,03 g
`2,0 g
`om g
`il!i tit
`ill 1[,
`100 mL
`8,15
`91.97%
`
`C ,f-IJ'J )t,~-I- 80 ~~<t.-:1ok7:r:T?TI-'J?k.l\lJ&{rTO~<t,f-­
`)~~~<t.':1o k 7:r: T?TI-'J? kll!l\lJO)~lEtt:T-1r
`
`II.
`
`fAl;J:, r'20)c;i:)L)o)*II/i£~. m~~<t.·. I;.(rO):1ok7:r:T?T~
`
`6
`
`Page 20 of 613
`
`

`
`Jllii\lJ '" -
`I-'
`:;:foA7xT7TI' ~ c)A
`JJ<l'n!/&J
`;t;c)1ilt
`;t;c)ltY
`c) A
`J:![il:rZ;/-t!' Iv'" -
`7]< V '/IVrZ- I, 80
`TO ";-.Jt;J<
`Iv
`l1' ~ l:":='lv l:" P V 1-';/
`:r. 7 I-IiltT I- ~ c) A
`<lliJiilEIiltTI-V'1A
`JJ<i'IJ1il:T I, ~ '1.b.
`m1l'lJJ<
`:,£Jll;
`I pH
`60 °C • 4 il!!
`
`BF (PE)' (~IJ
`7':1')
`
`A-OI (PE)
`
`A·03 (PE)
`
`0.1 g
`
`l.lg
`l.lg
`0.005 g
`0.15 g
`.
`2.0 g
`0.Q2 g
`0.2 g
`ii:llt
`iiJll;
`100mL
`8.20
`91.45%
`
`0.1 g
`
`l.lg
`l.lg
`0.005 g
`.
`0.Q2 g
`2.0g
`0,02 g
`.
`iiit
`ii:!il:
`100mL
`8.20
`93.61%
`
`0.1 g
`
`I.lg
`l.lg
`0.005 g
`.
`0.03 g
`2.0 g
`0.02 g
`.
`lim
`iiHl
`100mL
`8.27
`95.07%
`
`12.
`
`1'AI;J:'I'lIf"(!, :11J~I? I- P2002B 116 l!I. v: P2002B 131 O)-!\IcV(
`
`2 ;:;h. i;,~ ~c r PEJ 1;J:g;:)E 11m~~ fJl7f-1 ) IT V /!gn~~ -:>"( i'Tp;h. b::.c~~
`,,*l..* .. 0
`
`7
`
`Page 21 of 613
`
`

`
`J-'
`lJl:Ii'lrJ =z
`290 ¥,-, 131 ¥,-, 813 ¥,-
`&U' 606 ~Wf (7)M
`2,3-1 C 3-2~I::ftt2(7)
`::J-fC
`7'0 A7 "'-TlIT ~ ~
`<7 A 71< fer!ll)!
`*?AA'
`*<7$
`J.j[ l~--z /-If)v =z '" ? A
`;f, ~ '/ )V--z- ~ 80
`7- P ~-~ij~-/i,..-
`,r ~ 1::' _ )v 1::'0 ~ J-' /'
`OJ: 7' ~ 1\Ii-j- ~ ~ <7 j,
`i!H: M, AA' -j- I, ~ ? A
`7I<fiid~T I· ~ ? A
`i#\ll'Ji<
`~Bk
`I pH
`
`:107':11;7
`
`A-Ot
`
`A-02
`
`1!tt
`
`0.1 g
`
`l.l g
`l.l g
`0,005 g
`0.15 g
`
`2.0 g
`0.02 g
`0.2 g
`jtJ;jJJt
`jiiJ lli;
`100 mL'
`8.3
`
`A-04 & 11 3-t
`II
`
`A-OS & 11 3-2
`II
`
`0,1 g
`
`l.lg
`l.lg
`0,005 g
`-
`0.02 g
`2.0 g
`0,02g
`-
`iliii,t
`il\jjj;
`100 mL
`8.19
`
`0.1 g
`
`l.lg
`1.1 g
`0.005 g
`-
`0.05 g
`2.0 g
`0.02 g
`-
`.i&lik
`jgllli;
`100 mL
`8.20
`
`1 iili It,!,iijj B 1'1<1:, ~HJ!IlI(;!I~ Q) :lltil!'~ lJl:IlfrJ 1 "I) I) " f- )~iI"> t= L) Q) *J!1iX;~Q) m1'
`~:~l n '""T o ;:: Q)Ji!~lJl:I1!'J 100 "1)1) "f-)~~t=L)Q)*g1iX;~Q).I::iI!:!il '(
`,'""To
`
`8
`
`Page 22 of 613
`
`

`
`13.
`
`f,(~:::J-" P2002B 116 O)!'i::~~O)-n;C'''''N )~,,-~ 80 ~;%;t':I
`
`[J.L. 7I1-o71-~IJ? .L.~J1lJ (:I [J 1-';107 clliJ~ ) O)1*ff~:Il:h O)l;t~f.JI'fl!¥C'~1ffii
`
`~:I1.*Lf=o (iilfHHli Bo) 'fL.1;J:, l;t~~:::J-" P2002B131 O)~~O)-I!l-C;T[J
`
`~"t",-)~~;%<t;2-:JO):I[J.L.7I1-o71-~IJ ?.L.~J1lJ(290~, 131 ~, 813 ~
`
`&tF 606 ~t~lIf0)!/l2~" !/l3-1 ~c!/l3-2 ~O)!MJ1lJ A-04c A-05)0)1*ff~
`
`:hO)~~~1ffiiHT:b-tt*Lf=o (~I>t.~ Co) ~:I1.';O)~1!'JO)1*ff~:hr.t~
`
`I;J:, 3-[J ';1 1 \~F.i1J ( rEP J) O)~'I'~~m L 'C 'fl/fC'i'f:b:l1., ~41;J:, ~O)lit~
`
`f.J'';~';:I1.t=*1i~~~atL*Lt= o (~I;j'~lIi'i B &tF Co)
`
`14.
`
`EP O)l!i'l'I::J:.Qc, aj(~~:I1..QIl!J1lJl;J:At-';11 \-O)-:JL ,t=iIlIGOO~:I1.
`
`t= l;t~>1i 'lrl' 1:::$1 ;l~:I1. *T 0 ~!£ ~ 0) Illil ::if.>f= 'J, 'II-~OOli I;J: lO':::J [J::,(cid:173)
`
`ff%ltlll-III (CFU) ImL O)l!l!iOOill, 'll-A 1E111;J: 1 0' CFUlmL 0) rill iOO ill f.JW-ir.~:I1.
`
`*Lf=o 'II-~J1lJ~ ~IJ k O)lit~'lfl::, • .w;~Jlf.JI~OOI;J: 10' CFUlmL, .OOI;J:
`
`10' CFUlmL c>:i:.QJ:? -t:l1.-f:l1.ll!I~:I1.*Lf=o Illl1Jt.li(~'lrl'I;J: 20 f.J'';
`
`25 °c C'1*ff~:I1.*Lf=o 'II-~'lff.J'';. 6l11frdl. 24I11fr.,. 1 ~r.'. 2 ~r.'. 3 ~
`r.,& u: 4 iIllr.'1JtI::"t:,-:1 )~f.Jlj*IIl!~:I1.*Lf=o 'II-"t:"-:1 )~f.J''; 0.5 mL 0)"t:"-:1
`
`)~f.J14.5 mL O)iIlIGlE!£I!:t!:I1i.;~-C;lIHft~:I1.*Lt= o 10 f:'llIHftf.J< I f.J''; 3 @lff
`
`:b:l1.. -tL 'C:II1-iIft;i>;~ 1 mL f.J(:If... - H::ffi[f.J':I1.*Lt=. J!:I:: 15 f.J''; 20 mL 0)
`
`j~lt!Jf.J('II-:If...-H=JlI;l';:I1.*Lt=o ~il!iIMil ::JI!I L 'C I;J:. 1'i!ftJ1lJ(O.1 %f...
`
`9
`
`Page 23 of 613
`
`

`
`f,lt~'io/J
`Staphylococcus aureus A Tee
`6538
`Escherichia coli ATeC 8739
`Pseudomonas aeruginosa A Tee
`9027
`Candida afbicans A TCC 1023 1
`Aspergillus niger ATCC 16404
`
`IMU'i4
`
`30 - 35°C
`
`20-25°C
`
`Ir:IBM
`
`Aiii
`
`~;tt';(])f!ilffl\iJ niit~IJ' ';14I.;;tt t~*6*I;l:H~(])c<t:>')"C.'9 (i~1t
`15.
`.'In B & v: C):
`
`4IlI!81lt (CFUlmL)
`
`1§l1l6
`lill24 1f1l7 11I1I14 11I'121 11I1I 28
`~rJllf ~n.'lf Blf
`Blf
`Blf
`Blf
`4.0 x
`3. 1 x
`7.0 x
`10'
`\0'
`10'
`2.5 x
`10'
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`-
`-
`
`0
`
`-
`
`-
`
`10
`
`0
`
`0
`
`4.7 x
`10'
`6.0 x
`10'
`
`0
`3.0 x
`10'
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`~r:rT'Y
`'7
`
`l§lllllt
`
`S. aureus
`
`E. coli
`
`P.
`aeruginos
`a
`C.
`albicans
`
`A. niger
`
`4.3 x
`10'
`6.9 x
`10'
`
`\.Ix
`10'
`
`3.2 x
`10'
`1. 1 x
`10'
`
`Page 24 of613
`
`

`
`fl3-1
`A-OJ (1lIl
`.--~-,\-
`04)
`
`S. allreus
`
`E. coli
`
`P.
`aeruginosa
`e
`albicans
`
`A. niger
`
`.3-2
`A-02
`(1lIl • • 1:
`A-04 )
`
`S. aurells
`
`E. coli
`
`P.
`aerllginos
`a
`e
`albicans
`
`A. niger
`
`Illlil!(
`
`2.1 x
`10'
`6.5 x
`10'
`5.8 x
`10'
`3.2 x
`10'
`1.8 x
`10'
`
`Itllil!(
`
`2.1 x
`10'
`6.5 x
`10'
`5.8 x
`10'
`3.2 x
`10'
`1.8 x
`10'
`
`*BiAilil!( (CFU/mL)
`
`Illl6 Illl24 Ill17 111114 1l1l21 Ill128
`~Mlf ~lUIlf Bli Blf
`B !~ Blf
`3.0 x
`10'
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`-
`-
`
`0
`
`0
`-
`-
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`*WAilil!( (CFUlmL)
`
`1l1l6 Illl24 Illl7 111114 1l1l21 1l1l2B
`Blf
`Blf
`~r.'lf ~r.'lf Blf Blf
`2.0 x
`1.7 x
`10'
`10'
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`-
`
`-
`
`0
`
`0
`
`-
`
`-
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`11
`
`Page 25 of613
`
`

`
`il!ill'iA
`
`~i\! B
`
`3-[J';JJ ~!lJOl;l;&*
`~m(s. aureus, E. coli &
`P. oeruginosa)O)!EJ.lllt Am (G. albicans & A.
`tJljlllll."., 6I1iir.9, 24
`niger) O)!til!i~tJqllll
`Iliir.9, &LF 28 Sitl::i-
`tJ,o;, 7 Sit, 11100 ~XF
`111 000 ~XF, *t=I;J:I§!t!J ~tJ( *' ~ (\i,{, ~ '1;J:j;t 1-',
`ni'n 11100 1;l1',
`1::~1-'L, tJ,~, I;litm
`'F OJIiIl*-r:;JiH' 0
`mm(s. aureus, E. coli &
`Jl:M(G. alhicans &A.
`P. aerugjnosa) O)1:~liI:
`niger) 0) !t !II ~tJqll!ItJ'
`tJlJllfltJ'o;, 24 11iiM, 7
`0;, 14 Sitl:: 1110 ~XF I::
`Sitl::i-ni'n 1/ 10 I;l
`l!l1>L, fJ'~, ~),itOO~
`1', *t=l;J: 111000 1;l1'
`1::~1>, tJ,~, ~),ifOO~ tJ(*,~(\i,{,~ ";J:~1-' ,
`fJI*, ~(\i, {,~ '1;J:iJ;t 1> 0
`
`12
`
`Page 26 of 613
`
`

`
`E. E93-$iH1#0)?—5!L‘£I.$1!IE‘§.‘=i|:l‘;1'5J7'1'=é‘:h.T:-‘r'—§
`
`18.
`13.
`
`290 ~, 131 ~, 813 ~ &1t 606 ~11j,'F0)~ 1, ~ 2 &1t~ 3 1i0)
`290 v'%\ 131%. 813 -%-ELU? 606 %?$r.=¥F0)§% 1. % 2 RUEE 3 £0)
`
`§!fiIJ*\03$A0)F£éii:‘+1:§'5a wet. #%‘r§‘£I:§afi:<#rL7*:$t§?—9io<‘E0)ct5I:
`
`Lts1:1a£é>h.7‘:25\1:<>L\t1AB’a7‘.;$u§fiEfil,tL~a:=f° ififiiifi A. B Rafi C
`
`°&§an‘)r:2l:E§%1:T<a‘=#rL7‘:§fiIJ*(>EfC§§-;‘=~—/31(290 1%‘. 131%‘ 813 %)3‘zU
`
`606 %-4%*rE¢0>§E 1. % 2. 221%’ 3 i1:'.3:afla‘=#mrL\é§~!§IJ0>J+f;Ed‘) «MA
`
`0)BE15l:§'5é?. flxlat. E0)ot5l:l,‘C%0)J:57fa21fi$E7fi$fi£E?rLf:75\U)1l§JAE4]
`
`7‘.;5Eu§%iE7ELtL\i3‘o
`
`19.
`
`$1113. :_:1:1 7.=“J1:e9%EIa)f%§IE$0)F11|Ji320>TI:._t1:iI&r:
`
`.:t7ffiE%I‘IEEfi?i:"). $11EEwiuéfifilcgfiéfiéhfziitimld:$'CEa€Fi>
`
`U. '|fi$Ei& TEEl:§13§7‘.ié<#’L7‘:{#E5$li%'CE%‘C‘$;?§ :‘:{‘§Li:‘:>#’Lé:¢‘:’&EE
`
`L,$fi‘o
`
`ft ~~ir
`
`if
`
`~.!R
`
`B1'1":
`
`'/6 , OJ , I?
`
`13
`
`Page 27 of 613
`Page 2.7 of 613
`
`

`
`APPENDIX A
`
`APPENDIX AAPPENDIX A
`
`Page 28 of 613
`
`Page 28 of 613Page 28 of 613
`
`

`
`I
`
`CE RTIFICATION OJ.' TKA NSL ATIO N
`
`The undersigned. Ryan Malefio. whose adJrt.'s.~ is JflLt2~ tf....rAlu~6.i- dcdal't'S
`nn{j ~lal':S <t~ full ows:
`/.on~fo,.. (eA"--~
`
`I am well acquaint~d \~ilh the Engl ish and Japa l1~sc lang.uagt: ~: r Imv\: in the past translated
`numerous J ~lpa.ncse dOClllllcnts onegal and/or technical ~ontcnt into Ellg.lish.
`
`I hallc been requested to translate inTO English the aUached Jap:U1c~c documents titled:
`
`•
`
`ExhIbit A_ I~OOOBl77 data relied on.pdf
`
`• Exhibit R_P2002D1l6 data relied on.pdf
`
`• Exhibit C_P2002B131 data relied on.pdf
`
`To cnpics of Ulese Japanese documents I thereforl' attach the English translations and my
`Certification of Tnlnslation.
`
`I hereby certify that the English lmnsJations of the allUchcd documents titled
`
`• Exhibit A_P2000BJ77 data relied on .pdf
`
`• Exhibit B_P2002BI16 data relied on.pdf
`
`• EIhibit C_P2002B131 data relied on .pdf
`
`ru't. 10 lhe best of my knowledge and ability. accurate ImnslatiOIlS.
`
`And [ dl.'Clare fllrlher tha! all statements made herein of III)' own k.nowledge are true, that HI1
`statements made on information and belief:;re believed co be true. and Ihat falSe statements and tile like are
`plIllishubJ ,/, y fine and im risonmeni, or both, under Section 100! orTitle 18 of the United States Code.
`
`-74=~~~SJ:.
`
`!UV if ,. d-tJ1 L
`
`Date
`
`Page 29 of 613
`
`

`
`FORM Protocol 4-2- 1 (Version 3, I February 2000)
`
`Tcst Protocol
`
`Name of test: Study of the Formulation ofBronuck Ophthalmic Solution at pH 7
`Test code: P2000B 177
`Test system: None
`Development code: AHR 102828
`Test start date: 7 December 2000
`Scheduled stan date of test operations: 7 December 2000
`Scheduled end date of test operations: 15 March 2000 [sic]
`Scheduled test end date: 30 March 2001
`Test facility: Kobe Creative Ccnter, Senju Phannaceutical Co., Ltd.
`1-5-4 Murotani, Nishi -ku, Kobe-shi
`
`(Division of work duties)
`
`Study director: Shirou Sawa
`Study personnel:
`Test substance: Bromfenac sodium
`
`Purpose: Bromfenac sod ium is less soluble and unstable in the low pH range, so the pH (m idpoint of the
`standard) of Bronuck Ophthalmic Solution is sel to 8.3 . The pH of tears is general ly said to be around 7 to
`7.4, and since the pH of Bronuck Ophthalmic Solution is believed to be near the upper limit used in
`ophthalmic solutions, a fo rmulation at a lower pH is desired. Bromfenac sodium has an acetic acid group
`in its molecules, so its solubility increases at a pH of 6.5 and higher. Since the dissolution of Bronuck
`Ophthalmic Solution mostly occurs in association with the acetic acid group, control of the acetic acid
`group for solu bilization and stabilization is believed to be important. Although the addition of counterions
`to controllhe acetic acid group has been considered, bromfenac sod ium form s insoluble com plexes due 10
`the addition of quaternary ammon ium salt and becomes cloudy. Thus, the purpose of Ihis test is to usc
`water-soluble aminosugars to study the sol ubilization and stabilization of bromfenac sodium, even if
`complexes are fonned.
`
`Test method:
`1) Solubilization study
`Add an excessive amount of bromfenac sodium to acetic acid (pH 3--6), phosphoric acid (pH 5-7), or
`boric acid (pH 7-9) buffer sol ution, add 0.1 to 1.0% ofN-methylglucamine or glucosamine hydrochloride,
`and adj ust the pH with hydrochloric acid. Filter this solution, and measure Ihe concentration ofbromfenac
`sodium in the filtrate by HPLC.
`
`J
`
`Page 30 of 613
`
`

`
`2) Stabilization study
`the following bromfenac sodium
`to
`Add N-methylglucamine or glucosamine hydrochloride
`ophthalmic solutions, and adjust the pH to 7. In experiment 1), the N-mcthylglucamine or glucosamine
`hydr