Attorney Docket No.:PPT-3004-US-PR1
`
`PATENT
`
`IN 'I‘HE UNITED STATES PATENT AND TRADEMARK OFFICE
`
`FILING UNDER 37 C.F.R.l.53(C)
`
`to
`
`.
`
` o Assistant Commissioner for Patents
`T 3 Box Provisional Patent Application
`Washington, D.C. 2023]
`
`Express Mail Label No. E,144§32;§ I 13113
`Date of Deposit July 12, 1922
`
`Sir:
`
`This is a request for filing a provisional application under 37 C.F.R. l.53(c), of the
`
`following inventors:
`
`Larsen, Bjarne Due, a Danish Citizen, residing at at Arildsgard 5, 1.th, DK—2700 Brtzmshcij
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`Denmark; and
`
`Mikkelsen, Jens Damsgaard a Danish Citizen, residing at Borgevej 9, 2800 Lyngby Denmark
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`for the application entitled “NOVEL PEPTTDES THAT LOWER BLOOD GLUCOSE LEVELS”.
`
`The provisional application contains:
`
`Q2 pages of specification
`
`1 sheets of drawings
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`Address all future communications to Cheryl H. Agris, Ph.D., Attorney at Law, P.O. Box
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`806, Pelham, N.Y. 10803.
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`Enclosed is a check for the filing fee of $75. The assignee, Zealand Pharmaceuticals N8 is a
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`small entity. A duplicate of this sheet is enclosed.
`
`Respectfully submitted,
`
`I
`/' 3*-'r‘~,
`
`t/;"‘f}71[
`
`Date:
`
`Cheryl
`
`.
`
`Attorney at Law
`P.O. Box 806
`
`Pelham, N.Y. 10803
`
`(914) 712-0093
`
`SANOFI-AVENTIS Exhibit 1014 - Page i
`IPR for Patent No. 8,951 ,962
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`

`
`09-07-99
`
`15 :5i
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`Fra-ZEALAND PHA~ACEUTICALS A/S
`
`+4545161375
`
`Attorney Docket No.: PPT~3004-US-PRl
`
`PATENT
`
`IN THE UNITED STATES PATENT A."'l'D TRADEMARK OFFICE
`
`In re: Application of; Larsen et al.
`
`Serial No.: To Be Decided
`
`Group Art Unit; To be Decided
`
`Filed: Concurrently Herewith
`
`Examiner: To Be Decided
`
`For: NOVEL PEPTIDES THAT LOWER BLOOD GLUCOSE LEVELS
`
`VERIFlED STATEMENT CLAIMING SMALL ENTITY STATUS
`(37 CPR l.9(f) and l.27(c))-Small Business Concern
`
`Assistant Commissioner for Patents
`Washington, DC 20231
`
`Sir:
`
`I hereby declare that I am
`
`[ ] the owner of the small business concern identified below:
`( x) an official of the small business concern empovvered to act on behalf of the
`concern identified below:
`
`Name of Concern ........ Zealand Pharl1laceuticals A/S .............. ..
`Address of Concern ., .... Agern Alle 3 ........................ ., ...... ..
`............. , ............... DK-2970 H0:rsholm.Derunark ............. .
`
`I hereby declare that the above-identified small business concern qualifies as a small business
`concern as defined in 13 CFR 1:21.3-18, and reproduced in 37 CFR 2.9(d), for purposes of paying
`reduced fees under section 41(a) and (b) of Title 35, United States Code, in that the number of
`employees ofthe concern, including those of its affiliates, does not exceed 500 persons. For
`
`SANOFI-AVENTIS Exhibit 1014 - Page ii
`
`IPR for Patent No. 8,951,962
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`

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`C9-07·99
`
`15:52
`
`F ra·ZEALAN~ PHARMACEUTICALS A/S
`
`+45451$ 1375
`
`T-129
`
`S.13/17
`
`F·2ST
`
`purposes of this statement, (I) the munber of employees of the business concern is the average
`over the previous fiscal year of the concern of the persons employed on a full-time, part-time or
`temporary basis during each of the pay periods of the fiscal year, and (2) concerns are affiliates of
`each ol11er when either dil'ectly orindirectly, one concern controls or has the power to conttol the
`other, or a third party or parties controls or has the power to control both.
`
`I hereby declare that rights under contract or law have been conveyed to and remain with the
`small business concern and/or there is an obligation under contract or law by the inventor(s) to
`convey rights to the small business concern with regard to the invention
`
`entitled ....... NOVEL PEPTIDES THAT LOWER BLOOD GLUCOSE LEVELS .................. .
`
`by inventor(s) ..... Bjame Due Larsen .. and Jens Damsgaard Mikkelsen .............................. .
`described in
`
`[ x] the specification filed herewith
`[ ] application serial no ................ > filed ... " ..................................... .
`[ ] parent no ............ ~issued ........................................................ ..
`
`SANOFI-AVENTIS Exhibit 1014 - Page iii
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`IPR for Patent No. 8,951,962
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`

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`09-07-99
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`15:53
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`F ra·Z5ALAND PHARMACEUT I CA~S A/S
`
`+4545161975
`
`T-129
`
`S.14/1T
`
`F-261
`
`If the rights held by the above-identified small business concern are not exclusive, each
`individual, concern or organization having rights to the invention is listed below* and no rights
`to rhe invenhon are held by any person~ other than the inventor, who could not qualify as a small
`business concern under 3 7 CFR L9(d) or by any concern which would not qualify as a small
`business concern under 3 7 CFR 1. 9( d) or a nonprofit organi2:ation under 3 7 CFR 1. 9( e).
`
`FULL NAME ........................................................................ ..
`ADDRESS .............................................................................. .
`
`[ 1 INDIVIDUAL [] SMALL BUSINESS CONCER.N [ ] NONPROFIT
`ORGANIZATION
`
`FULL NAME .......................................................................... .
`. <\DDRESS .............................................................................. .
`
`[) INDIVIDUAL [] SMALL BUSINESS CONCERN []NONPROFIT
`ORGANIZATION
`
`FULL NAME ....................................................................... .,.
`ADDRESS ............................................................................. .
`
`[] INDIVIDUAL [ l SMALL BUSINESS CONCE&'\l []NONPROFIT
`ORGANIZATION
`
`FULL NAME ........................................................................ ..
`ADDRESS ............................................................................. .
`
`[] INDIVJDUAL [] SMALL BUSINESS CONCERN []NONPROFIT
`
`ORGANIZATION
`
`SANOFI-AVENTIS Exhibit 1014 - Page iv
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`IPR for Patent No. 8,951,962
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`

`
`09-07-99
`
`15 :5S
`
`Fri-ZEALAND PHARMACEUTICA~S A/S
`
`+4545161375
`
`T-129
`
`s.15/17
`
`F~zer
`
`I acknowledge the duty to file, in this application or patent, notification of any change in status
`
`resulting in loss of entitlement to small entity status prior to paying, or at the time of paying, the
`
`earliest of the issue fee or any maintenance fee due after the date on which status as a small entity
`
`is no louger appropriate. (37CFR 1.28(b))
`
`I hereby declare that all statements made herein of my own knowledge are true a:nd that all
`statements made on infonnation and belief are believed to be true; and fwther that these
`statements were made with the knowledge that willful false statements and the like so made are
`
`punishable by fine or imprisonment, or both, under section l 00 l of Title 18 of the United States
`Code, and that such willful false statements may jeopardize the validity of the application, any
`
`patent issuing thereon, or any patent to which this verified statement is directed.
`
`Send correspondence to:
`CHERYL H. AGRlS, PH.D.
`Attorney at Law
`P.O. Box 806
`Pelham, N.Y. 10803
`
`Direct Telephone calls to:
`CHERYL H. AGRIS, PH.D.
`Atton1ey at Law
`(914) 712-0093
`
`S1' S l tV E'~S
`~ Vf\
`.
`.
`Naxne of person S1g1ung ....................................................... .
`Title of person other than owner .......... h..~···t····'-': .. f..9. ...... .
`Address of person signing .. Agern Alle 3 .................................... .
`.......... .... ........ ..... .. .. ... DK-2970 H0rsholm.Deruna:rk .............. ..
`
`• 0 'f 0 . . . I . • O P~. f • 0 .,_ • • I •
`
`I 4 . I.,_ I I
`
`I .. I • • I I I 0 I • • 0 I 0 • • I I . 0 10 0<0 0 0 0 WI 0 • • 0 I
`
`I WY I 0 ':zo ... I 0 t ... orr • I 0 • • o
`
`<if~ q n
`.
`<:::rlf
`S1gnature .... 7'L..~ .. ~f· Date, ..................... :-1
`
`* NOTE: Separate verified statements are required from each
`named persoc, concern or organization having rights to the
`invention averring to their status as small entities.
`(37 CFR l.27)
`
`SANOFI-AVENTIS Exhibit 1014 - Page v
`
`IPR for Patent No. 8,951,962
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`

`
`PPT~3004-US—PRl
`
`PATENT
`
`NOVEL PEPTIDES THAT LOWER BLOOD GLUCOSE LEVELS
`
`5
`
`FIELD OF THE INVENTION
`
`The present invention relates to novel variants that lower blood glucose levels, specific
`
`variants of exendin-4. The invention further relates to peptide conjugates that lower
`
`blood glucose levels and which have increased oral bioavailability.
`
`10
`
`BACKGROUND OF THE INVENTION
`
`
`
`
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`A number of hormones that lower blood glucose levels are released from the
`
`gastrointestinal mucosa in response to the presence and absorption of nutrients in the gut.
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`15
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`These include gastrin, secretin, g1ucose—dependentinsulinotropic polypeptide (GIP) and
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`glucagon-like peptide-1 (GLP—l). The most potent substance known is GLP-1 (C/lrskov,
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`1992, Diabetologia 35:701-71 1).
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`Glucagon—1ike peptide 1 (GLP-1) is a product of proglucagon, a 180 amino acid peptide
`
`20
`
`(Drucker, 1998, Diabetes 47: 1 59-169). The overall sequence of proglucagon contains the
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`29-amino acid sequence of glucagon, the 36 or 37 amino acid sequence of GLP-1 and the
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`34 amino acid sequence of glucagon-like peptide-2 (GLP-2), an intestinotrophic peptide.
`
`GLP-l has a number of functions. It is a physiological hormone that enhances the effect
`
`25
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`on insulin secretion in normal humans and is therefore an incretin hormone. This explains
`
`in part the augmented insulin response after oral versus intravenous glucose
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`administration. In addition, GLP-1 also lowers glucagon concentrations, slows gastric
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`SANOFI-AVENTIS Exhibit 1014 - Page 1
`IPR for Patent No. 8,951,962
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`

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`emptying, stimulates (pro )insulin biosynthesis, and enhances insulin sensitivity (Nauck,
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`1997, Horm. Metab. Res. 47:1253~1258). The peptide also enhances the ability for the
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`~-cells to sense and respond to glucose in subjects with impaired glucose tolerance
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`(Byrne, 1998, Eur. J. Clin. Invest. 28:72-78). The insulinotropic effect ofGLP-1 in
`
`5
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`humans increases the rate of glucose disappearance partly because of increased insulin
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`levels and partly because of enhanced insulin sensitivity (D'Alessio, 1994, Eur. J. Clin.
`
`Invest. 28:72-78). This has placed GLP-1 as a promising agent for treatment for type II
`
`diabetes. Active fragments ofGLP-1 have been found to be GLP-1(7-36) and GLP-1(7-
`
`37). However, a major pharmacological problem with native GLP-1 is its short half-life.
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`10
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`In humans and rats, GLP-1 is rapidly degraded by dipeptidyl peptidase-IV (DPP-IV) into
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`GLP-1(9-36)amide, acting as an endogenous GLP-1 receptor antagonist (Deacon, 1998,
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`Diabetologia 41 :271-278). Several strategies circumventing this problem have been
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`proposed, some using inhibitors of DPP-IV and others DPP-IV resistant analogues of
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`GLP-1(7-36)amide (Deacon, 1998, Diabetologia 41:271-278; Deacon et al., 1998,
`
`15 Diabetes 47:764-769; Ritzel, 1998, J. Endocrinol. 159:93-102; U.S. Patent No. 5,545,618;
`
`Pederson, 1998, Diabetes 47:1253-1258).
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`Exendins, another group of peptides that lower blood glucose levels have some sequence
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`similarity (53%) to GLP-1[7-36]NH2 (Goke et al., 1993, J. Biol. Chern. 268:19650-55).
`
`20
`
`The exendins are found in the venom of Helodermatidae or beaded lizards (Raufman,
`
`1996, Reg. Peptides 61:1-18). Exendin-3 is present in the venom of Heloderma
`
`horridum, the Mexican beaded lizard and exendin-4 is present in the venom of Heloderma
`
`suspectum, the Gila monster. Exendin-4 differs from exendin-3 at just positions two and
`
`three. The eDNA encoding the exendin-4 precursor protein, a 47 amino acid peptide
`
`25
`
`fused to the amino terminus of exendin-4 has been cloned and sequenced (Pohl et al.,
`
`1998, J. Biol. Chern. 273:9778-9784 and W098/35033).
`
`2
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`SANOFI-AVENTIS Exhibit 1014 - Page 2
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`IPR for Patent No. 8,951,962
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`Both exendin-3 and exendin-4 stimulate an increase in cellular cAMP production in guinea
`
`pig pancreatic acinar cells by interacting with exendin receptors (Raufman, 1996, Reg.
`
`Peptides 61: 1-18). Exendin-3 causes a biphasic increase in cellular cAMP production, but
`
`a monophasic increase in amylase release in pancreatic acinar cells. In contrast, exendin-4
`
`5
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`causes a monophasic increase in cAMP production and does not alter amylase release.
`
`Exendin-4 is a strong GLP-1 receptor agonist on isolated rat insulinoma cells (Goke et al.,
`
`1993, J. Biol. Chern. 268: 19650-55). This is expected as the domain of GLP-1 recognised
`
`by DPP-IV is not present in exendin-4 (Goke et al., 1993, J. Biol. Chern. 268:19650-55).
`
`J 0 Binding of e25I]GLP-1 to the nucleus of the solitary tract was inhibited concentration(cid:173)
`
`dependently by unlabelled GLP-1 and [Tyr39]exendin-4 with Ki values of3.5 and 9.4
`
`nM respectively, and similar values are found in cell lines (Goke et al., 1995, Eur. J.
`
`Neurosci. 7:2294-2300 and Goke et al., 1993, J. Bioi. Chern. 268:19650-55). Further,
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`exendin-4 given systemically lowers blood glucose levels by 40% in db/db mice
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`15
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`(W099/07404). Recently, Grieg et al. (1999, Diabetologia 42:45-SO)has shown a long
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`lasting blood glucose lowering effect of once daily intraperitoneal injection of exendin-4 to
`
`diabetic oblob mice).
`
`The use of exendin-3, exendin-4 and exendin agonists has been proposed for the treatment
`
`20
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`of diabetes mellitus, reducing gastric motility and delaying gastric emptying and the
`
`prevention of hyperglycemia (U.S. Patent No. 5,424,286, W098/05351
`
`andW099/07404) as well as for the reduction of food intake (W098/30231 ).
`
`OBJECTIVE OF THE INVENTION
`
`25
`
`There is a need for compounds that lower blood glucose levels in mammals, are stable and
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`effective when administered orally. Therefore, it is an objective of the invention to
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`provide novel compounds that lower blood glucose levels in mammals.
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`3
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`SANOFI-AVENTIS Exhibit 1014 - Page 3
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`IPR for Patent No. 8,951,962
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`SUMMARY OF THE INVENTION
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`The invention is directed to a novel variant of a parent exendin, wherein said parent
`
`exendin has an amino acid sequence having at least an 90% homology to exendin-4 and
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`5 wherein said variant lowers the blood glucose level in a mammal, binds to a GLP-1
`
`receptor and has at least one modification selected from the group consisting of (a)
`
`between one and five deletions at positions 34-39, and (b) contains a Lys at position 40
`
`having a lipophilic substituent attached to the epsilon amino group of said lysine.
`
`10
`
`Furthermore, the invention is directed to a peptide conjugate comprising a peptide X that
`
`lowers the blood glucose level in a mammal and binds to a GLP-1 receptor, wherein X is
`
`said variant, an exendin having at least 90% homology to exendin-4, or GLP-1 (7-36) or
`
`GLP-1 (7-37) having at least one modification selected from the group consisting of: (i)
`
`substitution of D-alanine, serine, glycine or alpha-amino isobutyric acid for alanine at
`
`15
`
`position 8 and (ii) a lipophilic substituent and Z, a peptide sequence, of 4-20 amino acid
`
`units covalently bound to said variant, wherein each amino acid unit in said peptide
`
`sequence, Z is selected from the group consisting of Ala, Leu, Ser, Thr, Tyr, Asn, Gin,
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`Asp, Glu, Lys, Arg, His, Met, Om, and amino acid units ofthe general formula I
`
`20
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`(I)
`
`wherein R1 and R2 are selected from the group consisting of hydrogen, C1_6-alkyl, phenyl,
`and phenyl-methyl, wherein c1·6-alkyl is optionally substituted with from one to three
`
`substituents selected from halogen, hydroxy, amino, cyano, nitro, sulfone, and carboxy,
`
`25
`
`and phenyl and phenyl-methyl is optionally substituted with from one to three
`
`substituents selected from cl-6-alkyl, c2-6-alkenyl, halogen, hydroxy, amino, cyano, nitro,
`sulfono, and carboxy, or R1 and R2 together with the carbon atom to which they are
`
`4
`
`SANOFI-AVENTIS Exhibit 1014 - Page 4
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`IPR for Patent No. 8,951,962
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`

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`bound form a cyclopentyl, cyclohexyl, or cycloheptyl ring, e. g. 2,4-diaminobutanoic acid
`
`and 2,3-diarninopropanoic acid; and
`
`wherein the ratio between the minimum effective oral dose of said peptide conjugate and
`
`5
`
`the minimum effective oral dose of the peptide, X is at least 1:5.
`
`“:;;:ia
`
`ail.
`
`.:t:i;:=_..
`iifiilt
`3:5,
`
`
`
`The present invention also relates to a composition, e.g., a pharmaceutical composition,
`
`comprising said exendin variant or said peptide conjugate and a physiologically acceptable
`
`carrier, to said exendin variant of said peptide conjugate for use in therapy, a method of
`
`10
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`treating a disorder and to the use of said exendin variant or said peptide conjugate for the
`
`manufacture of a pharmaceutical composition for use in therapy. Specifically, the
`
`invention is directed to a method for stimulating insulin release in a mammal comprising
`
`administering an effective insulinotropic amount of the exendin variant or peptide
`
`conjugate of the present invention, a method of lowering blood glucose level in a mammal
`
`15
`
`comprising administering an amount of the exendin variant or peptide conjugate of the
`
`present invention effective to lower blood glucose level in said mammal, a method of
`
`reducing gastric motility in a mammal in an amount of the exendin variant or peptide
`
`conjugate of the present invention effective to reduce gastric motility, a method of
`
`delaying gastric emptying in a mammal in an amount of the exendin variant or peptide
`
`20
`
`conjugate of the present invention effective to delay gastric emptying, a method of
`
`inhibiting food uptake in a mammal in an amount of the exendin variant or peptide
`
`conjugate of the present invention effective to inhibit food uptake and a method of
`
`lowering plasma lipid level in a mammal comprising administering an amount of the
`
`exendin variant or peptide conjugate of the present invention effective to lower plasma
`
`25
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`lipid level in said mammal.
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`SANOFI-AVENTIS Exhibit 1014 - Page 5
`IPR for Patent No. 8,951,962
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`

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`Specifically, the exendin variant or peptide conjugate of the present invention may be
`
`used in treatment of diabetes type 1 or type 2, obesity, eating disorders, bone disease,
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`hyperglycemia, metabolic disorders, bone disease, gastric disease and insulin resistance
`
`syndrome.
`
`The present invention also relates to methods for the preparation of said exendin variant
`
`or said peptide conjugate, by means of recombinant DNA technology comprising the
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`steps of (a) introducing a nucleic acid sequence encoding said variant or conjugate into a
`
`host cell and (b) culturing said host cell and (c) isolating said variant or conjugate from the
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`10
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`culture or (a) culturing a recombinant host cell comprising a nucleic acid sequence
`
`encoding said variant or conjugate under conditions permitting the production of said
`
`variant or conjugate and (b) isolating said variant or conjugate from the culture.
`
`15
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`The method also relates to methods for the preparation of said peptide conjugate in which
`
`peptide X is obtained via recombinant DNA methods by isolating said peptide. X is then
`
`conjugated to Z which is attached to a solid support or has been prepared by solid phase
`
`synthetic methods. Furthermore, the invention relates to the preparation of the peptide
`
`conjugate of the present invention by peptide synthetic methods. Furthennore, the
`
`invention relates to the preparation of the peptide conjugate of the present invention by
`
`20
`
`peptide synthetic methods.
`
`Furthermore,
`
`the invention relates to the use of a peptide sequence (Z)
`
`for
`
`the
`
`preparation of said peptide conjugate.
`
`SANOFI-AVENTIS Exhibit 1014 - Page 6
`IPR for Patent No. 8,951,962
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`

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`BRIEF DESCRIPTION OF THE FIGURES
`
`Figure 1 shows the effect of ZS42-0009 (des Pro36-exendin-4-NH3) on blood glucose
`
`levels of mice.
`
`Figure 2 shows the effect of ZS42-0010 (des Pro”.-exendin-4-Lys5-NH; on the blood
`
`glucose levels of mice.
`
`Figure 3 shows the effect of zs42-0o19 (Glys Lys37(palmitoyl)-GLPl—(7-36)(Huma.n)-
`
`LYS7-NH2 on the blood glucose levels of mice.
`
`DETAILED DESCRIPTION OF THE INVENTION
`
`Exendin Variants
`
`The exendin variant of the present invention is a variant of a parent exendin peptide
`
`having at least about 90% homology and most preferably at least about 95% to exendin-4,
`
`which have exendin activity, e.g., lowers the blood glucose level in a mammal and binds to
`
`a GLP-1 receptor. In a preferred embodiment, the parent exendin peptide has an amino
`
`acid sequence which differs by five amino acids, preferably by four amino acids, more
`
`preferably by three amino acids, even more preferably by two amino acids from the
`
`amino acid sequence of exendin—4.
`
`The homology referred to above of the parent exendin is determined as the degree of
`
`identity between two protein sequences indicating a derivation of the first sequence from
`
`the second. The homology may suitably be determined by means of computer programs
`
`SANOFI-AVENTIS Exhibit 1014 - Page 7
`IPR for Patent No. 8,951,962
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`known in the art such as GAP proided in the GCG program package (Program Manual for
`
`the Wisconsin Package, Version 8, August 1994, Genetics Computer Group, 575 Science
`
`Drive, Madison, Wisconsin, USA 53711) (Needleman, S.B. and Wunsch, C.D., (1970), J.
`
`Mol. Biol. 48:443—453). The following settings for polypeptide sequence comparison
`
`5
`
`may be used: GAP creation penalty of 3.0 and GAP extension penalty of 0.1.
`
`In one embodiment, the exendin variant comprises between one and five deletions at
`
`positions 34-39.
`
`10
`
`In another embodiment, the variant comprises an additional residue at position 40, a
`
`lysine residue which comprises a lipophilic substituent bound to the epsilon amino group
`
`of lysine via an amide bond. The lipophilic substituent may be the acyl group of a
`
`straight-chain or branched fatty acid or a straight-chain or branched alkane
`
`ot,co-—dicarboxylic acid . The acyl group may have the formula CH3(CH2),,CO-, wherein n
`
`15
`
`is an integer from 4-38 and preferably from 4-24. In a specific embodiment, the acyl
`group is selected from the group consisting of CH3(CH2)6CO-, CH3(CH2)gCO-,
`
`Cl-l3(CH2)1oCO~, CH3(C}:l2)12CO-, CH3(CH2)14CO-, CH3(CH2)16CO-, CH3(CH2)13CO-,
`
`CH3(CH2)2oCO-, and CH3(CH2)22CO-. The acyl group may have the formula
`
`HOOC(CH2)mCO-, wherein n is an integer from 4-38 and preferably from 4-24. In a
`
`20
`
`specific embodiment, the acyl group is selected from the group consisting of
`
`gas
`
`f?*;
`
`
`
`HOOC(CH2)14CO-, HOOC(CH2)16C0-, HOOC(CH2),3CO-, HOOC(CH2)20CO- and
`
`HOOC(CH2)22CO-. In a more specific embodiment, the lipophilic substituent is selected
`
`from the group consisting of tetradecanoyl, to-carboxynonadccanoyl, 7-deoxycholoyl,
`
`choloyl, palmitoyl and Iithocholyl. In a most specific embodiment, the lipophilic
`
`25
`
`substituent is palmitoyl.
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`SANOFI-AVENTIS Exhibit 1014 - Page 8
`IPR for Patent No. 8,951 ,962
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`

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`Alternatively, the liphophilic substituent may have an NH group. Specific embodiments
`
`include but are not limited to the formulae
`
`CH3(CH2)a((CH2)bCOOH)CHNHCO(CH2)2CO-, in which a and b are integers and a+b is
`
`an integer of from 8 to 33, preferably from 12 to 28; CH3(CH2)cC0NHCH(COOl-I)
`
`(CH2)2CO-, wherein c is an integer of from 10 to 24; CH3(CH2)d CONI-ICH(CH2)2
`
`(COOH)CO-, wherein d is an integer of from 8 to 24; COOH(CH2)eCO-, wherein e is an
`
`integer of from 8 to 24; -NHCH(COOH)(CH2)4NHCO(CH3),CH3, wherein f is an integer
`
`of from 8 to 18; -NHCH(COOH)(CH2)4NHCOCH(CHz)zC0OH)NHC0(CH2)gCH3,
`
`wherein g is an integer of from 10 to 16; -NHCH(COOH)(CH2)4NHCO(CH2)2
`
`CH(CO0H)NHC.O(CH3),,CH3, wherein h is an integer of 0 or from 1 to 22 and
`
`preferably from 10 to 16.
`
`The variant may be in a most specific embodiment selected fiom the group consisting of
`
`des Ser”-exendin—4-NH2, des Pro”-exendin-4-NH2, des Ala”-exendin-4—NH2 and des
`
`Gly3“-exendin—4~NH2, des Sex”-(Lys4° (palmitoyl))exendin-4-NH2, des Gly“-(Lys4°
`
`(pa.lmitoyl))exendin-4-NH2, des Ala35-(Lys4° (pa1mitoyl))exendin—4-NI-I2 and des Pro”-
`
`(Lys"° (pa1mitoy1))exendin-4-NH2.
`
`Peptide Conjugates
`
`The peptide sequence X used in the peptide conjugate of the present invention lowers the
`
`blood glucose level of a mammal as determined by assays known in the art for a particular
`
`peptide. Examples of such an assay are described herein.
`
`In the present context, the peptide sequence X may be the exendin variant or parent
`
`exendin described above. Alternatively, X may be GLP-l (7-36) or GLP-1 (7-37) having
`
`at least one modification selected from the group consisting of a substitution of D-alanine,
`
`serine, glycine or alpha-arnino isobutyric acid for alanine at position 8 or any of the
`
`SANOFI-AVENTIS Exhibit 1014 - Page 9
`IPR for Patent No. 8,951,962
`
`

`
`lipophilic substituents described above. In a specific embodiment, the lipophilic
`
`substituent is attached to the epsilon amino group of a lysine residue. In a more specific
`
`embodiment, the lipophilic substituent is attached to the epsilon amino group of the
`
`lysine residue(s) at position 26, 34 and/or 37. In a most specific embodiment, X is
`
`5 Gly 8Lys37N-palmitoyl-GLP-l (7-36), Gly8 -GLP-1 (7-36), Gly8Lys34N-palmitoyl-GLP-
`
`1 (7-36) or Gly 8Lys2~-palmitoyl-GLP-l (7-36).
`
`Z is typically a peptide sequence of 4-20 amino acid residues, e.g., in the range of 4-15,
`
`more preferably in the range of 4-10 in particular in the range of 4-7 amino acid residues,
`
`10
`
`e.g., of 4, 5, 6 or 7 amino acid residues. Each of the amino acid residues in the peptide
`
`sequence Z are independently selected from Ala, Leu, Ser, Thr, Tyr, Asn, Gln, Asp, Glu,
`
`Lys, Arg, His, Met, Om, and amino acids ofthe formula I as defined herein such as
`
`diaminobutanoic acid or diaminopropanoic acid. Preferably, the amino acid residues are
`
`selected from Ser, Thr, Tyr, Asn, Gln, Asp, Glu, Lys, Arg, His, Om, and Met, more
`
`15
`
`preferably from Glu, Lys, and Met, especially Lys. The above-mentioned amino acids
`
`may have either D- or L-configuration, but preferably the above-mentioned amino acids
`
`have an L-configuration.
`
`Thus, illustrative examples of the peptide sequence Z are:
`
`20
`
`Lys-Lys-Lys-Lys, Xaa-Lys-Lys-Lys, Lys-Xaa-Lys-Lys, Lys-Lys-Xaa-Lys, Lys-Lys(cid:173)
`
`Lys-Xaa, Xaa-Xaa-Lys-Lys, Xaa-Lys-Xaa-Lys, Xaa-Lys-Lys-Xaa, Lys-Xaa-Xaa-Lys,
`
`Lys-Xaa-Lys-Xaa, Lys-Lys-Xaa-Xaa, Xaa-Xaa-Xaa-Lys, Xaa-Xaa-Lys-Xaa, Xaa-Lys(cid:173)
`
`Xaa-Xaa, Lys-Xaa-Xaa-Xaa, Xaa-Xaa-Xaa-Xaa, Lys-Lys-Lys-Lys-Lys, Xaa-Lys-Lys(cid:173)
`
`Lys-Lys, Lys-Xaa-Lys-Lys-Lys, Lys-Lys-Xaa-Lys-Lys, Lys-Lys-Lys-Xaa-Lys, Lys-
`
`25
`
`Lys-Lys-Lys-Xaa, Xaa-Xaa-Lys-Lys-Lys, Xaa-Lys-Xaa-Lys-Lys, Xaa-Lys-Lys-Xaa(cid:173)
`
`Lys, Xaa-Lys-Lys-Lys-Xaa, Lys-Xaa-Xaa-Lys-Lys, Lys-Xaa-Lys-Xaa-Lys, Lys-Xaa(cid:173)
`
`Lys-Lys-Xaa, Lys-Lys-Xaa-Xaa-Lys, Lys-Lys-Xaa-Lys-Xaa, Lys-Lys-Lys-Xaa-Xaa,
`
`10
`
`SANOFI-AVENTIS Exhibit 1014 - Page 10
`
`IPR for Patent No. 8,951,962
`
`

`
`Lys-Lys-Xaa-Xaa-Xaa, Lys-Xaa-Lys-Xaa-Xaa, Lys-Xaa-Xaa-Lys-Xaa, Lys-Xaa-Xaa(cid:173)
`
`Xaa-Lys, Xaa-Lys-Lys-Xaa-Xaa, Xaa-Lys-Xaa-Xaa-Lys, Xaa-Xaa-Lys-Lys-Xaa, Xaa(cid:173)
`
`Xaa-Lys-Xaa-Lys, Xaa-Xaa-Xaa-Lys-Lys, Lys-Xaa-Xaa-Xaa-Xaa, Xaa-Lys-Xaa-Xaa(cid:173)
`
`Xaa, Xaa-Xaa-Lys-Xaa-Xaa, Xaa-Xaa-Xaa-Lys-Xaa, Xaa-Xaa-Xaa-Xaa-Lys, Xaa-Xaa-
`
`5 Xaa-Xaa-Xaa, Lys-Lys-Lys-Lys-Lys-Lys, Xaa-Lys-Lys-Lys-Lys-Lys, Lys-Xaa-Lys(cid:173)
`
`Lys-Lys-Lys, Lys-Lys-Xaa-Lys-Lys-Lys, Lys-Lys-Lys-Xaa-Lys-Lys, Lys-Lys-Lys(cid:173)
`
`Lys-Xaa-Lys, Lys-Lys-Lys-Lys-Lys-Xaa, Xaa-Xaa-Lys-Lys-Lys-Lys, Xaa-Lys-Xaa(cid:173)
`
`Lys-Lys-Lys, Xaa-Lys-Lys-Xaa-Lys-Lys, Xaa-Lys-Lys-Lys-Xaa-Lys, Xaa-Lys-Lys(cid:173)
`
`Lys-Lys-Xaa, Lys-Xaa-Xaa-Lys-Lys-Lys, Lys-Xaa-Lys-Xaa-Lys-Lys, Lys-Xaa-Lys-
`
`10
`
`Lys-Xaa-Lys, Lys-Xaa-Lys-Lys-Lys-Xaa, Lys-Lys-Xaa-Xaa-Lys-Lys, Lys-Lys-Xaa(cid:173)
`
`Lys-Xaa-Lys, Lys-Lys-Xaa-Lys-Lys-Xaa, Lys-Lys-Lys-Xaa-Xaa-Lys, Lys-Lys-Lys(cid:173)
`
`Xaa-Lys-Xaa, Lys-Lys-Lys-Lys-Xaa-Xaa, Xaa-Xaa-Xaa-Lys-Lys-Lys, Xaa-Xaa-Lys(cid:173)
`
`Xaa-Lys-Lys, Xaa-Xaa-Lys-Lys-Xaa-Lys, Xaa-Xaa-Lys-Lys-Lys-Xaa, Xaa-Lys-Xaa(cid:173)
`
`Xaa-Lys-Lys, Xaa-Lys-Xaa-Lys-Xaa-Lys, Xaa-Lys-Xaa-Lys-Lys-Xaa, Xaa-Lys-Lys-
`
`15 Xaa-Xaa-Lys, Xaa-Lys-Lys-Xaa-Lys-Xaa, Xaa-Lys-Lys-Lys-Xaa-Xaa, Lys-Lys-Lys(cid:173)
`
`Xaa-Xaa-Xaa, Lys-Lys-Xaa-Lys-Xaa-Xaa, Lys-Lys-Xaa-Xaa-Lys-Xaa, Lys-Lys-Xaa(cid:173)
`
`Xaa-Xaa-Lys, Lys-Xaa-Lys-Lys-Xaa-Xaa, Lys-Xaa-Lys-Xaa-Lys-Xaa, Lys-Xaa-Lys(cid:173)
`
`Xaa-Xaa-Lys, Lys-Xaa-Xaa-Lys-Lys-Xaa, Lys-Xaa-Xaa-Lys-Xaa-Lys, Lys-Xaa-Xaa(cid:173)
`
`Xaa-Lys-Lys, Lys-Lys-Xaa-Xaa-Xaa-Xaa, Lys-Xaa-Lys-Xaa-Xaa-Xaa, Lys-Xaa-Xaa-
`
`20
`
`Lys-Xaa-Xaa-Lys, Lys-Xaa-Xaa-Xaa-Lys-Xaa-Lys, Lys-Xaa-Xaa-Xaa-Xaa-Lys-Lys,
`
`Xaa-Lys-Lys-Xaa-Xaa-Xaa, Xaa-Lys-Xaa-Lys-Xaa-Xaa, Xaa-Lys-Xaa-Xaa-Lys-Xaa,
`
`Xaa-Lys-Xaa-Xaa-Xaa-Lys, Xaa-Xaa-Lys-Lys-Xaa-Xaa, Xaa-Xaa-Lys-Xaa-Lys-Xaa,
`
`Xaa-Xaa-Lys-Xaa-Xaa-Lys, Xaa-Xaa-Xaa-Lys-Lys-Xaa, Xaa-Xaa-Xaa-Lys-Xaa-Lys,
`
`Xaa-Xaa-Xaa-Xaa-Lys-Lys, Lys-Xaa-Xaa-Xaa-Xaa-Xaa, Xaa-Lys-Xaa-Xaa-Xaa-Xaa,
`
`25 Xaa-Xaa-Lys-Xaa-Xaa-Xaa, Xaa-Xaa-Xaa-Lys-Xaa-Xaa, Xaa-Xaa-Xaa-Xaa-Lys-Xaa,
`
`Xaa-Xaa-Xaa-Xaa-Xaa-Lys, Xaa-Xaa-Xaa-Xaa-Xaa-Xaa, Lys-Lys-Lys-Lys-Lys-Lys(cid:173)
`
`Lys, Xaa-Lys-Lys-Lys-Lys-Lys-Lys, Lys-Xaa-Lys-Lys-Lys-Lys-Lys, Lys-Lys-Xaa-
`
`11
`
`SANOFI-AVENTIS Exhibit 1014 - Page 11
`
`IPR for Patent No. 8,951,962
`
`

`
`Lys-Lys-Lys-Lys, Lys-Lys—Lys-Xaa-Lys-Lys-Lys, Lys-Lys-Lys-Lys~Xaa-Lys-Lys,
`
`Lys-Lys-Lys-Lys-Lys-Xaa—Lys, Lys-Lys-Lys-Lys-Lys-Lys—Xaa, Xaa~Xaa-Lys-Lys-
`
`Lys-Lys-Lys, Xaa-Lys-Xaa~Lys-Lys-Lys—Lys, Xaa-Lys~Lys-Xaa-Lys-Lys-Lys, Xaa-
`
`Lys-Lys-Lys-Xaa-Lys-Lys, Xaa-Lys-Lys-Lys-Lys-Xaa-Lys, Lys-Xaa-Xaa-Lys-Lys~
`
`5
`
`Lys-Lys, Lys-Xaa-Lys-Xaa-Lys-Lys-Lys, Lys-Xaa—Lys-Lys-Xaa-Lys~Lys, Lys-Xaa-
`
`Lys-Lys-Lys-Xaa-Lys, Lys-Lys-Xaa-Xaa-Lys-Lys-Lys, Lys-Lys-Xaa-Lys-Xaa-Lys-
`
`Lys, Lys-Lys-Xaa-Lys-Lys-Xaa-Lys, Lys-Lys-Lys-Xaa-Xaa-Lys-Lys, Lys-Lys-Lys-
`
`Xaa-Lys-Xaa-Lys, Lys-Lys-Lys-Lys-Xaa-Xaa-Lys, Xaa-Xaa-Xaa-Lys-Lys-Lys-Lys,
`
`Xaa-Xaa-Lys-Xaa-Lys-Lys-Lys, Xaa—Xaa-Lys-Lys-Xaa-Lys-Lys, Xaa-Xaa-Lys-Lys-
`
`10
`
`Lys-Xaa-Lys, Xaa-Lys-Xaa-Xaa—Lys-Lys-Lys, Xaa—Lys-Xaa-Lys-Xaa—Lys-Lys, Xaa-
`
`Lys-Xaa-Lys—Lys-Xaa-Lys, Xaa-Lys-Lys~Xaa—Xaa-Lys-Lys, Xaa—Lys-Lys-Xaa-Lys-
`
`Xaa-Lys, Xaa-Lys-Lys-Lys-Xaa-Lys-Xaa, Xaa-Lys-Lys-Xaa-Lys-Lys-Xaa, Xaa—Lys-
`
`Xaa—Lys- Lys-Lys—Xaa, Xaa—Lys-Lys-Lys-Xaa-Xaa-Lys, Lys—Xa.a-Lys-Lys-Lys-Xaa-
`
`Xaa, Xaa-Lys-Lys-Lys-Lys~Xaa-Xaa, Xaa-Lys-Lys-Lys-Xaa-Lys-Xaa, Xaa—Lys-Lys-
`
`15
`
`Lys-Xaa-Xaa-Lys, Lys-Lys—Lys-Lys-Xaa-Xaa-Xaa, Lys-Lys-Lys-Xaa~Xaa-Xaa-Lys,
`
`Lys-Lys-Lys-Xaa-Lys-Xaa-Xaa, Lys-Lys-Xaa-Lys-Lys-Xaa-Xaa, Lys-Lys-Xaa-Xaa-
`
`Lys—Xaa-Lys, Lys-Lys-Xaa—Xaa—Xaa-Lys-Lys, Lys—Lys~Xaa-Lys—Lys—Xaa-Xaa, Lys-
`
`Xaa-Lys-Lys-Xaa-Xaa-Lys, Lys-Xaa-Lys-Xaa-Lys-Xaa-Lys, Lys-Xaa-Lys-Xaa-Xaa-
`
`Lys-Lys, Lys-Xaa-Xaa-Lys-Lys-Xaa-Lys, Lys-Xaa-Xaa-Lys-Xaa-Lys-Lys, Lys-Xaa-
`
`20
`
`Xaa—Xaa-Lys~Lys-Lys, Lys-Lys—Xaa—Xaa-Xaa-Xaa-Lys, Lys—Xaa-Lys-Xaa-Xaa-Xaa-
`
`Lys, Lys-Xaa~Xaa-Lys-Xaa~Xaa-Lys, Lys-Xaa-Xaa—Xaa-Lys-Xaa-Lys, Lys-Xaa-Xaa-
`
`Xaa-Xaa-Lys-Lys, Xaa-Lys-Lys-Xaa-Xaa-Xaa-Lys, Xaa-Lys-Xaa-Lys-Xaa-Xaa-Lys,
`
`Xaa-Lys-Xaa-Xaa-Lys-Xaa~Lys, Xaa-Lys—Xaa—Xaa-Xaa-Lys-Lys, Xaa-Xaa-Lys-Lys-
`
`Xaa-Xaa-Lys, Xaa-Xaa-Lys-Xaa-Lys-Xaa-Lys, Xaa-Xaa-Lys-Xaa-Xaa-Lys—Lys, Xaa-
`
`25
`
`Xaa-Xaa-Lys-Lys-Xaa-Lys, Xaa-Xaa-Xaa-Lys-Xaa-Lys~Lys, Xaa-Xaa-Xaa-Xaa-Lys-
`
`Lys-Lys, Lys-Xaa-Xaa—Xaa~Xaa-Xaa-Lys, Xaa-Lys-Xaa-Xaa-Xaa-Xaa-Lys, Xaa-Xaa-
`
`Lys-Xaa-Xaa-Xaa-Lys, Xaa-Xaa-Xaa—Lys-Xaa-Xaa—Lys, Xaa-Xaa—Xaa-Xaa-Lys-Xaa-
`
`12
`
`
`
`
`
`SANOFI-AVENTIS Exhibit 1014 - Page 12
`IPR for Patent No. 8,951 ,962
`
`

`
`Lys, Xaa-Xaa-Xaa-Xaa—Xaa-Lys-Lys, Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Lys, Lys-Xaa-Xaa-
`
`Xaa-Xaa—Xaa-Xaa, Xaa-Xaa-Xaa-Xaa-Xaa-Lys-Xaa, Xaa-Lys-Xaa-Xaa-Xaa-Xaa-Xaa,
`
`Xaa-Xaa-Lys-Xaa-Xaa-Xaa, Xaa-Xaa-Xaa-Xaa-Lys-Xaa, Xaa-Xaa-Xaa-Xaa-Xaa-Xaa
`
`Xaa, wherein each Xaa is independently selected from the group consisting of Ala, Leu,
`
`5
`
`Ser, Thr, Tyr, Asn, Gln, Asp, Glu, Arg, His, Met, Om, and amino acids of the fonnula I
`
`as defined herein, e.g., Dbu or Dpr.
`
`As indicated above, the amino acid residues of Z may of course all be different or all be
`
`identical. However, in interesting embodiments of the present invention, the amino acid
`
`10
`
`residues in Z are selected from two or three different amino acids, or are identical amino
`
`acids. Examples of suitable peptide sequences, wherein the amino acid residues in Z are
`
`identical are e.g., (Lys),,, wherein n is an integer in the range from 4 to 15, preferably in
`
`the range from 4 to 10, such as in the range from 4 to 8, e.g., in the range from about 4 to
`
`7, e.g., Lys4, Lyss Lysg , LYS7. Examples of suitable peptide sequences, wherein the
`
`15
`
`amino acid residues in Z are selected from about two different amino acids are e.g., (Lys-
`
`Xaa)m or (Xaa-Lys)m, wherein m is an integer in the range from about 2 to 7, preferably in
`
`the range from 2 to 5, such as in the range from 2 to 4, e.g., 3, and Xaa is independently
`
`selected from the group consisting of Set, Thr, Tyr, Asn, Gln, Asp, Glu, Arg, His, Om,
`
`2,4-diaminobutanoic acid, 2,3-diaminopropanoic acid and Met. More preferably such
`
`20
`
`peptide sequences are e.g., (Lys-Xaa); or (Xaa-Lys)3, wherein Xaa is as defined above,
`
`f"="“.
`
`9;:
`
`
`
`such as (Lys—Glu)3 or (Glu~Lys)3. Other examples of suitable peptide sequences, wherein
`
`the amino acid residues in Z are selected from about two amino acid residues are e.g.,
`
`Lysp-Xaaq or Xaa‘,-Lysq, wherein p and q are integer