IN THE UNITED STA TES PA TENT AND TRADEMARK OFFICE
`
`U.S. Patent No.
`
`5,670,373
`
`Issued:
`
`September 23, 1997
`
`Inventors:
`
`Tadamitsu KISHIMOTO
`
`Applicant:
`
`Chugai Seiyaku Kabushiki Kaisha
`
`Product:
`
`Actemra® (tocilizumab), humanized anti-
`human IL-6R monoclonal antibody
`
`APPLICATION FOR PATENT TERM EXTENSION
`
`UNDER 35 U.S.C. §156
`
`WASH_6768488.2
`
`Trademark Office. Applicant is an agent of the patent owner and is authorized to act on
`behalf of Kishimoto.
`
`Mail Stop: Hatch Waxman PTE
`Commissioner for Patents
`PO. Box 1450
`
`Alexandria, VA 22313-1450
`
`Sir or Madam:
`
`Applicant Chugai Seiyaku Kabushiki Kaisha (“Applicant”) hereby applies for patent
`
`term extension of U.S. Patent No. 5,670,373 under 35 U.S.C. § 156(d) and 37 C.F.R. § 1.740.
`
`U.S. Patent No. 5,670,373 issued in the named of the sole inventor, Tadamitsu
`
`Kishimoto (“Kishimoto”). No assignment has been made or recorded in the U.S. Patent and
`
`This application is based on the approval by the United States Food and Drug
`
`Administration (“FDA”) of a Biologics License Application (BLA # 125276/0) on January 8,
`
`2010, for Actemra® (tocilizumab), a humanized monoclonal antibody that is an interleukin—6
`
`receptor (IL-6R) inhibitor, for the treatment of adult patients with rheumatoid arthritis.
`
`For convenience,
`
`the information contained in this application will be presented
`
`according to the format set forth in 37 C.F.R. § 1.740(a).
`
`03/02/2010 SSfiflbnkfl 00000004 5670373
`
`01 FC:1457
`02 FC:1999
`
`1120.00 GP
`90.00 DP
`
`Mylan v. Genentech
`IPR2016-00710
`Merck Ex. 1128, Pg. 1
`
`

`

`Atty. Dkt. No. 029099-0107
`
`(1)
`
`A Complete Identification Of The Approved Product As By Appropriate
`Chemical And Generic Name, Physical Structure Or Characteristics
`
`The approved product is a recombinant humanized anti-human Interleukin-6 Receptor
`
`(IL-6R) monoclonal
`
`antibody of the immunoglobulin IgG1
`
`subclass,
`
`formulated for
`
`intravenous (iv) infilsion. The approved product has the trade name Acternra®, and established
`
`name of “tocilizumab.”
`
`Actemra® (tocilizumab) is an IgGlK (gamma 1, kappa) antibody with a typical H2L2
`
`structure. The amino acid sequence of the light chain is shown in Figure 1.
`
`Figpre 1: Amino Acid Seguence of the L Chain
`
`DIQMTQSPSS LSASVGDRVT ITCRASQDIS SYLNWYQQKP GKAPKLLIYY
`
`50
`
`The amino acid sequence of the heavy chain is shown in Figure 2.
`
`Figpre 2: Amino Acid Seguence of the H Chain
`
`pEVQLQESGPG LVRPSQTLSL TCTVSGYSIT SDHAWSWVRQ PPGRGLEWIG 50
`YISYSGITTY
`NPSLKSRVTM
`100
`TAVYYCARSL
`LRDTSKNQFS
`LRLSSVTAADV
`STKGPSVFPL
`150
`TAALGCLVKD
`APSSKSTSGG
`LYSLSSVVTV
`
`ARTTAMDYWG
`
`YFPEPVTVSW
`
`ICNVNHKPSN
`
`QGSLVTVSSA
`NSGALTSGVH
`TKVDKKVEPK
`
`DTLMISRTPE
`
`VTCVVVDVSH
`
`TFPAVLQSSG
`SCDKTHTCPP
`EDPEVKFNWY
`
`CPAPELLGGP
`VDGVEVHNAK
`
`TYRVVSVLTV
`
`LHQDWLNGKE
`
`YKCKVSNKAL
`
`PAPIEKTISK
`
`YTLPPSRDEL
`DSDGSFFLYS
`
`TKNQVSLTCL
`KLTVDKSRWQ
`
`VKGFYPSDIA
`
`VEWESNGQPE
`
`QGNVFSCSVM
`
`HEALHNHYTQ
`
`KSLSLSPG
`
`TSRLHSGVPS RFSGSGSGTD FTFTISSLQP EDIATYYCQQ GNTLPYTFGQ 100
`GTKVEIKRTV AAPSVFIFPP SDEQLKSGTA SVVCLLNNFY PREAKVQWKV 150
`DNALQSGNSQ ESVTEQDSKD STYSLSSTLT LSKADYEKHK VYACEVTHQG 200
`LSSPVTKSFN RGEC
`214
`
`VVASH_67684882
`
`PSSSLGTQTY
`SVFLFPPKPK
`
`TKPREEQYNS
`
`AKGQPREPQV
`NNYKTTPPVL
`
`200
`
`250
`
`300
`
`350
`
`400
`
`448
`
`(2)
`
`A Complete Identification Of The Federal Statute Including The Applicable
`Provision Of Law Under Which The Regulatory Review Occurred
`
`Regulatory review of the approved product occurred under § 505(i) of the Federal
`
`Food, Drug, and Cosmetic Act (“FFDCA”), 21 U.S.C. § 355(i) (see also 21 C.F.R. Part 312)
`
`Merck Ex. 1128, Pg. 2
`
`

`

`Atty. Dkt. No. 029099-0107
`
`and § 351(a) of the Public Health Service Act (“PHSA”), 42 U.S.C. §262(a) (see also 21
`
`C.F.R. Part 314 and 601).
`
`(3)
`
`An Identification Of The Date On Which The Product Received Permission For
`
`Commercial Marketing 0r Use Under The Provision Of Law Under Which The
`Applicable Regulatory Review Period Occurred
`
`The approved product, Actemra® (tocilizumab), received permission for commercial
`
`marketing on Janan 8, 2010, when the FDA approved BLA # 125276/0 for the treatment of
`
`adults with rheumatoid arthritis, under § 351(a) of the PHSA.
`
`(4)
`
`In The Case Of A Drug Product, An Identification Of Each Active Ingredient In
`The Product And As To Each Active Ingredient, A Statement That It Has Not
`Been Previously Approved For Commercial Marketing Or Use Under The
`Federal Food, Drug And Cosmetic Act, The Public Health Service Act, Or The
`Virus—Serum—Toxin Act, Or A Statement Of When The Active Ingredient Was
`Approved For Commercial Marketing Or Use {Either Alone Or In Combination
`With Other Active Ingredients}, The Use For Which It Was Approved, And The
`Provision Of Law Under Which It Was Approved
`
`WASH_6768488.2
`
`The approved product contains only one active ingredient, tocilizumab. Tocilizumab
`
`(Actemra®) has not been previously approved for commercial marketing or use under the
`
`Federal Food, Drug, and Cosmetic Act, the Public Health Service Act, or the Virus-Serum-
`
`Toxin Act.
`
`(5)
`
`A Statement That The Application Is Being Submitted Within The Sixty, Day
`Period Permitted For Submission Pursuant To 37 C.F.R. § 1.7201!” And An
`Identification Of The Date Of The Last Day On Which The Application Could
`Be Submitted
`
`The Application is being submitted within the sixty day period permitted under
`
`37 C.F.R. § 1.720(t).
`
`The last day on which the application can be submitted is March 8 2010, i.e., sixty
`
`days from the date (Janum 8, 2010) the approved product first received permission for
`
`commercial marketing or use under the PHSA.
`
`Merck Ex. 1128, Pg. 3
`
`

`

`Atty. Dkt. No. 029099-0107
`
`(6)
`
`The patent for which an extension is being sought is:
`
`Inventors:
`
`Tadamitsu Kishimoto
`
`Patent No:
`
`US. Patent No. 5,670,373
`
`Title:
`
`Antibody to Human Interleukin-6 Receptor
`
`Issue Date:
`
`September 23, 1997
`
`Current Expiration Date:
`
`January 2, 2013
`
`A Complete Identification Of The Patent For Which An Extension Is Being
`Sought By The Name Of The Inventor, The Patent Number, The Date Of Issue,
`And The Date Of Expiration.
`
`WASH_6768488.2
`
`(the expiration date of Jan. 2, 2013 is based upon a terminal
`
`disclaimer over US 5,480,796, which expires on January 2,
`
`2013.)
`
`The ’373 patent has not previously been extended.
`
`(7)
`
`A Copy Of The Patent For Which An Extension Is Being Sought, Including The
`Entire Specification [Including Claims} And Drawings
`
`A copy of US. Patent No. 5,670,373 is attached hereto as Exhibit 1.
`
`A Copy Of Any Disclaimer, Certificate Of Correction, Receipt Of Maintenance
`Fee Payment, Or Reexamination Certificate Issued In The Patent.
`
`No certificates of correction or reexamination certificates have been issued in the
`
`patent. A terminal disclaimer, however, was filed over US Patent No. US 5,480,796, a copy
`
`of which is attached hereto as Exhibit 2. Copies of the 4th, 8th and 12th year maintenance fee
`
`payment receipts is attached hereto as Exhibit 3.
`
`Merck Ex. 1128, Pg. 4
`
`

`

`Actemra® (tocilizumab) is a monoclonal antibody that binds an interleukin-6 receptor
`
`and specifically, a human interleukin-6 receptor. Actemra® is approved for use in treating
`
`“adult patients with moderately - to severely — active rheumatoid arthritis who have had an
`
`inadequate response to one or more TNF antagonist
`
`therapies.”
`
`See Full Prescribing
`
`Information for Actemra®, section 1 (Exhibit 4). Rheumatoid arthritis is a chronic systemic
`
`disease primarily of the joints, marked by inflammatory changes in the synovial membranes
`
`and articular structures and by atrophy and rarefaction of the bones.
`
`US. Patent No. 5,670,373 claims an isolated antibody that specifically binds to human
`
`interleukin-6 receptor, which reads on the approved product Acternra®. Accordingly,
`
`applicable claims of the ’373 patent for patent term extension which claim the approved
`
`product include claims 1 and 3:
`
`1.
`
`An isolated antibody to human interleukin—6 receptor, wherein said antibody
`
`specifically binds to said human interleukin-6 receptor.
`
`An antibody according to claim 1, wherein said antibody is monoclonal.
`
`(9)
`
`Atty. Dkt. No. 029099-0107
`
`A Statement That The Patent Claims The Approved Product, Or A Method Of
`Using Or Manufacturing The Approved Producta And A Showing Which Lists
`Each Applicable Patent Claim And Demonstrates The Manner In Which At
`Least One Such Patent Claim Reads On The Approved Product
`
`WAS H_6768488.2
`
`Merck Ex. 1128, Pg. 5
`
`

`

`Atty. Dkt. No. 029099-0107
`
`IND Effective Date:
`
`November 4, 2004
`
`BB IND Number:
`
`1 1972
`
`The date on which a Biologic License Application (BLA) was initially submitted and
`the BLA number:
`
`BLA Submission Date:
`
`November 19, 2007
`
`BLA Number:
`
`125276/0
`
`The date on which the BLA was approved:
`
`BLA Approval Date:
`
`January 8, 2010
`
`(10)
`
`The relevant dates and information pursuant to 35 U.S.C. § 156(g) needed to enable
`
`the Secretary of Health and Human Services to determine the applicable regulatory review
`
`period for the Approved Product (a human biological product) are as follows:
`
`(A)
`
`The effective date of the Investigational New Drug Application (IND) and the IND
`number:
`-
`
`A Statement Beginning On A New Page Of The Relevant Dates And Information
`Pursuant To 35 U.S.C. 1561g) In Order To Enable The Secretagx Of Health And
`Human Services To Determine The Applicable Regulatogy Review Period
`‘
`
`WASH_6768488.2
`
`Merck Ex. 1128, Pg. 6
`
`

`

`(11)
`
`Please see attached Exhibit 5 for this Description. Several significant dates are also
`
`summarized below. Applicants reserve the right to supplement the activity description in
`
`Exhibit 5 if additional clarification is required.
`
`October 5, 2004
`
`FDA confirmation ofIND submission
`
`November 4, 2004
`
`IND effective date
`
`November 19, 2007
`
`BLA 1252 76/0 submitted to and received by FDA
`
`January 8, 2010
`
`FDA approves BLA 1252 76/0
`
`A Brief Description Beginning on a new Page of the Significant Activities
`Undertaken By The Marketing Agglicant During the Anglicable Regulatory
`Review Period With Respect To The Approved Product And The Significant
`Dates Applicable to Such Activities
`
`WASH_6768488.2
`
`Merck Ex. 1128, Pg. 7
`
`

`

`In the opinion of Applicant, US. Patent No. 5,670,373 is eligible for a patent term
`extension.
`
`The length of the patent term extension claimed is 1338 days, which is believed to
`
`extend the patent term to at least Sgptember 1I 2016.
`
`The length of the patent term extension claimed was determined in accordance with
`
`37 C.F.R. § 1.775, as the length of the regulatory review period for the Approved Product as
`
`defined in 37 C.F.R, § 1.775(0), reduced as appropriate pursuant to 37 C.F.R. § 1.775(d)(1)
`
`through (d)(6).
`
`As defined in 37 C.F.R. § 1.775(c), the length of the regulatory review period for the
`
`Approved Product is the sum of (1) and (2) below:
`
`(1) The number of days in the period beginning on the date an exemption under
`
`subsection (i) of section 505 or subsection (d) of section 507 of the Federal Food,
`
`Drug, and Cosmetic Act became effective for the approved product and ending
`
`on the date the application was initially submitted for such product under those
`
`sections or under section 351 of the Public Health Service Act.
`
`This is the number of days in the period beginning on the date the IND became
`
`effective, November 4 2004, and ending on the date the BLA was initially submitted,
`November-19 2007, which is 1111 days.
`
`(12)
`
`Atty. Dkt. No. 0290990107
`
`A Statement Beginning On A New Page That In The Opinion Of The Applicant
`The Patent Is Eligible For The Extension And A Statement As To The Length Of
`Extension Claimed, Including How The Length Of Extension Was Determined
`
`WAS H_6768488.2
`
`Merck Ex. 1128, Pg. 8
`
`

`

`Atty. Dkt. No. 029099—0107
`
`(2) The number of days in the period beginning on the date the application was
`
`initially submitted for the approved product under section 351 of the Public
`
`Health Service Act and ending on the date such application was approved under
`such section.
`
`This is the number of days in the period beginning on the date the BLA was initially
`
`submitted, November 19 2007, and ending on the date the BLA was approved, J21an 8,
`
`2_01_0, which is 782 days.
`
`The sum of(1) and (2) above is 11 11 days plus 782 days, which is 1893 days. Thus,
`
`the length of the regulatory review period for the Approved Product is 1893 days.
`
`In accordance with 37 C.F.R. § 1.775(d)(1), the length of the extension is initially
`determined as follows:
`
`W'AS H_6768488.2
`
`(i) Subtracting from the regulatory review period the number of days in
`
`periods (1) and (2) above which were on and before the date on which the
`
`patent issued.
`
`The IND became effective on November 4 2004, and the patent issued on September
`
`23, 1997. The number of days in periods (1) and (2) above which were on and before the date
`
`the patent issued is zero days. Subtracting 0 days from the regulatory review period leaves
`
`1893 days.
`
`(ii) Subtracting from the regulatory review period the number of days in
`
`periods (1) and (2) above-during which it is determined that Applicant did
`
`not act with due diligence.
`
`Applicant acted with due diligence during the entire regulatory review period. Thus,
`
`no deduction is required under this subsection.
`
`Merck Ex. 1128, Pg. 9
`
`

`

`Atty. Dkt. No. 029099-0107
`
`(iii) Subtracting from the regulatory review period one-half the number
`
`of days in (1) above remaining after that period is reduced in accordance
`
`with (i) and (ii) above.
`
`The number of days in period (1) is 1111 days. That number is reduced by 0 days, in
`
`accordance with (i) above. One half of 1111 days is 555 days. Subtracting 555 days from
`
`1893 days leaves 1338 days.
`
`Thus, in accordance with (d)(1), the length of the extension is
`
`(1893 days) — (0 days) — (555 days) = 338 days
`
`In accordance with 37 C.F.R. § 1.775(d)(2), the length of the extension is further
`
`determined as follows:
`
`(2) Adding
`
`the
`
`number
`
`of
`
`days
`
`determined
`
`in
`
`accordance with
`
`37 C.F.R. § l.775(d)(1) to the original term of the patent, as shortened by any
`terminal disclaimer.
`
`WASH_6768488.2
`
`The patent’s term is set to expire Janan 2, 2013 (a terminal disclaimer was filed over
`
`US Patent No. 5,480,796). 'The addition of 1338 days of Patent Term Extension to that term
`
`would extend the term to at least September 1, 2016.
`
`In accordance with 37 C.F.R. § 1.775(d)(3), the length of the extension is fithher
`
`determined as follows:
`
`(3) Adding 14 years to the date of approval of the application under section 351
`
`of the Public Health Service Act
`
`The application was approved on Janum 8a 2010. Adding 14 years to that date
`
`results in a date of J21an 8, 2024.
`
`In accordance with 37 C.F.R. § 1.775(d)(4), the length of the extension is further
`
`determined as follows:
`
`Merck Ex. 1128, Pg. 10
`
`

`

`Atty. Dkt. No. 029099-0107
`
`(4)
`
`By
`
`comparing
`
`the
`
`dates
`
`determined
`
`in
`
`accordance with
`
`37 C.F.R. § l.775(d)(2) and (d)(3) above and selecting the earlier date.
`
`The earlier date of the two dates determined above in accordance with 37 C.F.R.
`
`§ 1.776(d)(2) and (d)(3), is September 1, 2016.
`
`In accordance with 37 C.F.R. § l.775(d)(5), the length of the extension is further
`determined as follows:
`
`(5) If the original patent was issued after September 24, 1984, adding five years
`
`to the original expiration date and comparing that date to the date obtained in
`
`accordance with 37 C.F.R. § l.775(d)(4) and selecting the earlier date.
`
`The original term of the patent expires Jarmy 2, 2013.
`
`(a terminal disclaimer was
`
`filed over US Patent No. 5,480,796) Adding five years to Janum 2, 2013, results in a date of
`
`Januag 2, 2018.
`
`The earlier of Januag 2, 2018, and the date determined above in accordance with 37
`
`C.F.R. § 1.776(d)(4) is September 1, 2016.
`
`The provisions of 37 C.F.R.
`
`§
`
`l.775(d)(6) apply only to patents issued before
`
`September 24, 1984, and thus are not applicable here.
`
`Thus, the patent should be extended 1338 days which is believed to extend the term to
`
`at least September 1, 2016.
`
`WASH_6768488.2
`
`A Statement That The Applicant Acknowledges A Duty To Disclose To The
`Commissioner Of Patents And Trademarks And The Secretary Of Health And
`Human Services Or The Secretary Of Agriculture Any Information Which Is
`Material To The Determination Of Entitlement To The Extension Sought
`
`(13)
`
`Applicant acknowledges a duty to disclose to the Commissioner of Patents and
`
`Trademarks and the Secretary of Health and Human Services any information which is
`
`material to the determinations of entitlement to the extension sought herein.
`
`Merck Ex. 1128, Pg. 11
`
`

`

`Atty. Dkt. No. 029099—0107
`
`(14)
`
`The Prescribed Fee For Receiving And Acting Upon The Application For
`Extension
`
`A credit card authorization form for the prescribed fee is submitted herewith.
`
`Authorization is given to charge Deposit Account 19—0741 any deficiency in fees.
`
`(15)
`
`Inquiries and correspondence relating to this Application should be directed to the
`
`registered practitioner authorized to act on behalf of the patent owner in connection with this
`
`Application:
`
`The Name, Address, And Telephone Number Of The Person To Whom Inguiries
`And Correspondence Relating To The Application For Patent Term Extension
`Are To Be Directed
`
`WAS H_6768488.2
`
`Stephen B. Maebius
`Foley & Lardner LLP
`3000 K Street, NW.
`Washington, DC. 20007—5143
`Tel: 202—6726300
`
`(16)
`
`Certification Under 37 C.F.R. § 1.7401b2
`
`Two additional copies of this Application and Exhibits are submitted herewith in
`
`accordance with 37 C.F.R. § 1.740(b).
`
`The undersigned is a registered practitioner authorized to act on behalf of the patent
`
`owner in connection with this Application.
`
`Respectfiilly submitted,
`
`Date Februag 26, 2010
`
`By
`
`FOLEY & LARDNER LLP
`Foley & Lardner LLP
`3000 K Street, N.W.
`Washington, DC. 20007-5143
`Tel: 202-672-5300
`Facsimile: 202-672-5399
`
`Stephen B. Maebius
`Attorney for Applicant
`Registration No. 35,264
`
`Merck Ex. 1128, Pg. 12
`
`

`

`EXHIBIT 1
`
`US PATENT 5,670,373
`
`US PATENT 5,670,373 EXHIBIT 1
`
`Merck Ex. 1128, Pg. 13
`
`Merck Ex. 1128, Pg. 13
`
`

`

`.
`
`U8005670373A
`
`[11] Patent Number:
`
`[45] Date of Patent:
`
`5,670,373
`
`*Sep. 23, 1997
`
`Biological Abstracts vol. 28 . 1985 No. 65431.
`Biological Abstracts vol. 84 No. 12 1987 No. 118862.
`Nishikawa Chemical Abtracts. vol. 109 p. 193 No. 87407d
`Jun. 5. 1989.
`Yamasaki et a1. “Cloning and Expression of the Human
`Interleukin—6
`(BSF—2/IFNBZ)
`Receptor”.
`Science.
`24l:825——828 1988. Aug. 12).
`Kishimoto et al.. “Molecular Regulation Of 13 Lymphooyte
`Response”. Ann. Rev. Immunol, 6:485—512. 1988.
`Hirano et al.. “Human B—Cell Diifuentiation Factor Defined
`By An Anti—Peptide Antibody And Its Possible Role In
`Autoantibody Production". Pmc. Natl. Acad. Sci. USA,
`84:228—231.
`
`United States Patent
`Kishimoto
`
`[19]
`
`[54] ANTIBODY T0 HUMAN MERLEUKlN-G
`RECEPFOR
`
`[76]
`
`Inventor: Tadamitsu Kishimotn. 5—31.
`Nakanocho 3—chome. Tondabayashi.
`Osaka. Japan
`
`[‘1 Notice:
`
`The tam of this patent shall not extend
`beyond the expiration date of Pat. No.
`5.480.796.
`
`[21] Appl. No.: 357,080
`
`[22] Filed:
`
`Dec. 15, 1994
`
`Related US. Application Data
`
`[63] Continuation of Ser. No. 899,600, Jun. 18, 1992, abandoned,
`which is a continuation of Ser. No. $54,534, Jul. 20, 1990,
`abandoned. which is a continuatim-in-part of SCI: No.
`298.694. Jan. 19. 1989. Pat. No. 5,171,840.
`
`Foreign Application Priority Data
`[30]
`Jan. 22, 1988
`[JP]
`Japan ............................. 63-012387
`Jan. 25. 1988
`[JP]
`63-012599
`Aug. 4. 1988
`[JP]
`63-194855
`Jan. 14, 1989
`[JP]
`1-007461
`Jul. 20, 1989
`[JP]
`1-186016
`
`[51]
`
`Int. CL'5
`
`[52] 0.8. CI.
`
`[58] Field 6r Search
`
`CWK 16/00: C12? 21/04;
`C12N 15/00; C12N 5/00
`435/344: 435/7021; 435/172.2;
`435/240.27; 530388.23: 530,387.]
`435240.27. 172.2.
`435/7021. 344; 530/38823. 387.1
`
`[56]
`
`References Cited
`
`U.S. PATENT DOCUhJENTS
`5,171,840 12/1992 Kishimoto .............................. 530/350
`
`.
`.
`
`13 Claims, 5 Drawing Sheets
`
`Taga et al.. “Receptors Fcr B Cell Simulatory Factor 2
`Quantitation. Specificity. Distribution. and Regulation Of
`Their Expression”. J. Exp. Met/L, 166:967—981. 1987.
`Hirata et al.. J. Immunol. 143. 2900—2906. (1989 Nov. 1).
`Chemical Abstract vol. 109. 1988. p. 546. No. 717381).
`Deverwx et 111.. the Program Manual for the Sequence
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`Increased Expression Of BIA—137 mRNA In Tumor ” Sci.
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`Inducible By Growdx-Stimulatory“, Journal. 5:2529—2537
`Jul. 22. 1986.
`Hirano et al.. “Complementary DNA For A Novel Human
`Interleukin (BSF—Z) That Induces B Lymphocytes To Pro-
`duce Immunoglobulin". Nature, 324:73—76. Nov.. 1986.
`Hirano et a.. “Human B—Cell Difi'crentiation Factor Defined
`By An Anti—Peptide Antibody And Its Possible role In
`Autoantibody Production”. Prue. Natl. Acad. Sci. USA,
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`FOREIGN PATENT DOCUME‘TIS
`312996
`4/1989
`325474
`7/1989
`61-24697
`2/1986
`61-246197
`11/1986
`899776
`7/1990
`899774
`11/1990
`
`European Pat. 01f.
`European Pat. 011’.
`Japan
`quan
`Japan .
`Japan .
`ommz PUBLICATIONS
`
`188—189 and
`
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`Proceedings of the Japanese Society for Immunology.
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`ference of Japan Immunology (In Japanese).
`. Kishimoto et aL. Studies of B—Cell Growth. Difierentiation
`and Aberrant Control (Mar. 1989) (In Japanese).
`
`(List continued on next page.)
`
`Primary Examiner—Anthony C. Caputa
`Assistant Examiner—Mark Navarro
`Attorney Agent, or Finn—Foley & Lardner
`[57]
`ABSTRACT
`
`Antibodies, polyc10nal and monoclonal. which are capable
`of specifically binding to a human interleukin-6 receptor.
`Also included are monoclonal antibodies which competi-
`tively and non-competitively inhibit human interleukin-6.
`and a method of producing hybridornas of the said mono-
`clonal antibodies capable of specifically binding to a human
`interleukin-6 receptor.
`
`Merck Ex. 1128, Pg. 14
`
`

`

`UPI-[El PUBLICATIONS
`
`Yanmsnki et al. Science 241:825—828 Aug. 1988.
`
`Mikel ii et a.L pp. 3.1—3. 13 in Wei: et al.. Handbook of
`Experimental Immunology vol. 1 . 1936 . Blackwell.
`
`Milstein . pp. 107.1—107.12 in Wei: et 111. llka pf
`
`5,670,373
`Page 2
`
`Chu et al.. “SV40 DNA Transfecu’on Of Cells In Suspen-
`sion: Analysis Of The Efficiency Of Transcription And
`Translation Of T—Antigen". Gene, 13:197—202 1981.
`
`Fujii et aL J. Immunol. 137: 1552—1556. 1986.
`
`Hiram et a1. 1 Immunology 143(9) : 2900-2906 Nov. 1989.
`
`Tyndall et al.. "A Region Of The Polyoma ViflB Genome
`Between The Replication Origin And Late Protein”
`Research. 916231—6250 Nov. 23. 1981.
`Naknjima—lljima et 8.1., “Molecular Structure Of The Human
`Cytoplasmic B—aain Gene:
`Interspecies Homology Of
`Sequences In The muons". Proc. Natl. Acad. Sci. USA,
`82:6133—6137 Sep. 1985.
`Elis et 8.1.. “Replacement Of Insulin Receptor Tyrosine
`Residues 1162 And 1163 Compromises Insulin—Stimulated
`anase Activity And Uptake of 2—Deoxyglucos«:". Cell,
`45:721—732. Jun. 6. 1986.
`‘Wigler et a.. “Biochemical Transfer 0! Single—Copy
`Eucaryotic Genes Using Total Cellular DNA As Donor",
`Cell. 14:725—731Iu1. 1978‘
`
`Experimental Immunology vol. 4. 1986. Blackwell.
`
`Merck Ex. 1128, Pg. 15
`
`

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`US. Patent
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`Sep. 23, 1997
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`
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`
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`Merck Ex. 1128, Pg. 16
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`_U.S. Patent
`
`Sep. 23, 1997
`
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`Merck Ex. 1128, Pg. 17
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`Merck Ex. 1128, Pg. 19
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`

`

`US. Patent
`
`Sep. 23, 1997
`
`Sheet 5 of 5
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`5,670,373
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`Fig. 6(B)
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`INTENC ITY OF FLUORESCENCE
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`

`

`This application is a continuation applimtion of appli-
`cation Ser. No. 071899.600. filed Jun. 18. 1992. now
`abandoned, which is a continuation applimfion of Ser. No.
`(Tl/554.534. filed on Jul. 20. 1990. now abandoned, which is
`a continuation-impart application of Ser. No. 07/298594,
`filed .Ian. 19. 1989. now issued as US. Pat. No. 5.171.840.
`
`BACKGROUND OF THE INVENTION
`
`(1) Field of the Invention
`
`The present invention relates to antibodies which specifi-
`cally bind to a human 11,6 receptor. and a process for the
`preparation thaeof.
`(2) Description of the Related Art
`is a
`Intrz-leuldn-6 (hacinafter abtxeviated to “IL-6")
`Fotdn having various important physiological activities
`and participating broadly in the proliferation and differen-
`tiation of cells. Furthermore. it is reported that an abnormal
`production of lL-6 is possibly a factor causing various
`autoimmune diseases (see Kishimoto and leano. Ann. Rev.
`Immunol.. 6. page 485. 1988).
`L6 receptors on the cell memlxanc. which specifically
`bind to L6. were analyzed by Tag: et al.. and the number
`on each cell and binding constant to HAS were reported (see
`1. Exp. Med.. 196. page 967. 1987). Furthermore. the cDNA
`of human 11,6 receptor was isolated. and the primary
`structure thereof was reported by Yamazaki et al. (see
`Science. 241. page 825. 1988).
`lL-6 receptor which is
`prepared by a genetic engineaing method based on these
`results. is expected to become a therapeutic or diagnostic
`agetn for various immune diseases.
`For a mass production and homogeneous purification of
`such lL-6 receptors. the development of an antibody to the
`L6 receptor as a means for momptly identifying the 11.6
`receptor is required. but a monoclonal antibody capable of
`recognizing the IL-6 receptor has not been known.
`
`SUMMARY OF THE INVENTION
`
`Thuefa'e. a primary object of the present invention is to
`[rovide various types of antibodies to IL-6 receptor.
`To attain this object. the present invention provides anti-
`bodies to human interleukin-6 receptor, which is capable of
`binding specifically to a human interleukin-6 receptor.
`Flu-thermore. the present invention provides a hybridoma
`producing a monoclonal antibody having the above-
`mentioned propaties.
`The present invention also provides a process for the
`preparation of the hybridoma, which comprises immunizing
`a mammal with a human intaleukin-G receptor antigen.
`obtaining immunocytes from the mammal. fusing the immu-
`nocytes with a myeloma cells. and cloning a hybridoma cell
`line capable of recognizing a human interleukin-6 receptor
`from the fused cells.
`
`The present invention further provides a process for the
`preparation of an antibody to a human interleukin—6
`receptor. which comprises culturing the above—mentioned
`hybridoma and recovering a monoclonal antibody capable of
`recognizing a human interleukin-6 receptor from the culture.
`The present invention also provides a wooess for the
`preparation of a polyclonal antibody to a human
`interleukin—6 receptor. which comprises immunizing a mam-
`mal wiflr a human interleukin-6 receptor antigen and recov-
`
`2
`erlng a polyclonal antibody capable of recognizing a human
`interleukin-6 receptor from the immunized mammal.
`'
`
`BRIE7 DESCRIPTION OF THE DRAWINGS
`
`DESCRIPTION OF THE PREFERRED
`MODIMENTS
`
`Antibodies of the present invention specifically recognize
`a human 114-6 receptor. and include monoclonal and poly-
`clonal antibodies. The monoclonal antibodies include anti-
`bodies competitively inhibiting the binding of human 11,6 to
`a human IL-6 reeeptrx and antibodies not competitively
`inhibiting this binding. The former antibodies. include. for
`example. PMl monoclonal antibody poduced by the hybri-
`dorna PM] of the present invention. and the latter antibodies
`include. for example. MT18 monoclonal antibody produced
`by the hybridoma MT18.
`As an immunogen used for the preparation of the anti-
`bodies of the present invention. there can be mentioned
`animal cells having human lL-6 receptors expressed on the
`surface thereof. Such animal cells include a human-derived
`cell
`line producing human lL-6 receptm. for example.
`human myeloma cell line U266. and host cells transformed
`with DNA coding fw human lL-6 receptor. for example.
`animal cell line such as mouse T—cells transformed with a
`plasmid comprising cDNA coding for human IL-6 receptor.
`Nevertheless. these cell lines are not regarded as an effective
`antigen. because the quantity of the lL—6 receptor expressed
`on the cell surface is small.
`
`Nevertheless. even though such an immunogen is used to
`produce an antibody. once an antibody to a human IL-6
`receptor has been obtained. it is possible to prepare a more
`elfective immunogen by using this antibody. and to prepare
`
`1
`ANTIBODY 'DO HUMAN lNTERLEUKIN-6
`RECEIVIOR
`
`5,670,373
`
`FIG. lA—D shows the reactivity of a T~cell line Jurkat in
`which the 11/6 receptor is not expessed [ctn'ves (a) and (c)].
`or a Jurkat in which an L6 receptor cDNA-oontaining
`vector is introduced and the L6 receptor is durany
`expressed [curves (b) and (d)]. with the culture supernatant
`of MT18 [solid lines in curves (a) and (b)] or the culture
`supernatant of PMI [solid lines in curves (c) and (d)]. The
`dotted lines show the results for the mils not treated with the
`culture supa'natant;
`FIG. 2 shows the results obtained by solubilizing inta-
`nally labelled [1266 cells. efl'ecting immunoprecipitation
`with an MT18 monoclonal antibody (lane 1). WI mono-
`clonal antibody (lane 2), rabbit immunoglobul‘m (lane 3) or
`anti-peptide polyclonal antibody of Example 3 (lane 4). and
`subjecting same to SDS-polyaa'ylamide gel electrophoresis
`and autoradiography;
`FIG. 3 is a graph showing the inhibiting efl’eds of MI
`and M118 antibodies on the binding of lL—6 to the 11,6
`receptor according to the procedtn’e described in Example 4;
`FIG. 4A—B shows the results of a comparison of the
`bindings of the MT18 antibody [curve (a)] and PM] anti-
`body [ctn've (b)] to U266 cells in the presence (broken line)
`or absence (solid line) of lL-6. and the dotted lines show the ‘
`results for U266 cells treated with only ISL-6;
`FIG. 5 shows the incorporation of tritium-labelled thymi-
`dine by [(13 cells mltured with various concentrations of
`IL-6.-in the presence or absence of the mlture supernatant of
`MT18 or Phil; and
`FIG. 6A and B is a cell distribution diagram showing that
`the MT18 antibody binds only M receptor-producing
`cells.
`
`a Variety of antibodies by using this imrnuuogen. For
`
`Merck Ex. 1128, Pg. 21
`
`

`

`5,670,373
`
`4
`receptors expressed on the membrane sun-face was prepared
`as an immunogen. according to the following process.
`Namely. pBSF2R.236 and pSVZneo disclosed in the Japa-
`nese Patent Application No. 89-9774 were introduced into
`mouse T-cells (TILL—2 (A'I‘CC. ’I‘IBZM) according to a
`conventional method. and the screening was carried out
`according to a conventional method using G418. Finally. a
`cell line in which about 30.000 IL-6 receptors per cell were
`expressed was established and named “CI'BC3”.
`Immunization was carried out in the following manner.
`Namely. CI'BC3 was cultured by a conventional method
`using P10411640. washed with PBS buffer 4 times. and was
`intraperitoneally introduced to C57BLG mouse in a quantity
`of 1x107 cells per mouse once a Week. six times as a whole.
`The spleen cells from the immunized mouse were fused
`with myeloma cells P3Ul as a parent cell line. according to
`a conventional method using polyethylene glycol.
`The Screening was carried out in the following manner.
`Namely, pBSFRBtS and pSV2neo were introduced into
`human T‘cells JURKAT (A'I‘CC. CRL8163) negative to the
`L6 receptor. and am:- screening. a cell line in which about
`[00.000 lL—6 receptors per cell were expressed wa