a2) United States Patent
`US 6,284,727 Bl
`(10) Patent No.:
`Sep. 4, 2001
`(45) Date of Patent:
`Kim etal.
`
`US006284727B1
`
`(54) PROLONGED DELIVERY OF PEPTIDES
`
`OTHER PUBLICATIONS
`
`(75)
`
`Inventors: Yesook Kim; William J. Lambert;
`Hong Qi; Robert A Gelfand; Kieran
`F. Geoghegan; Dennis E. Danley, all
`of New York, NY (US)
`
`(73) Assignee: Scios, Inc., Sunnyvale, CA (US)
`
`(21) Appl. No.: 08/472,349
`
`(22)
`
`Filed:
`
`Jun. 7, 1995
`
`Related U.S. Application Data
`
`(63) Continuation of application No. 08/181,655,filed on Jan. 25,
`1994, now abandoned, which is a continuation-in-part of
`application No. 08/044,133, filed on Apr. 7, 1993, now
`abandoned.
`
`TMC0 eeeceeeecccssssessssseeccecessuneecseeseeee A61K 38/00
`CSL)
`(52) US. C0. eee eessseseecsecnecncceneesneesses 514/12; 530/324
`(58) Field of Search ieee 514/12; 530/324
`
`(56)
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`Kreymannet al, Laneef Dec. 5, 1987 p. 1300.*
`Conn’s Current Therapy p. 483. (1989).*
`Remington’s Pharmaceutical Sciences, 16th ed. p. 1555,
`1467, 1453, 1451, 1450, 1448, CH 91, 1980.*
`Crosset al., “Subcutaneous Absorption Kinetics And Local
`Tissue Distribution Of Interferon And Other Solutes,” J.
`Pharm. Pharmacol., 1993, 45, pp. 606-609.
`(*) Notice:—Subject to any disclaimer, the term ofthis
`Hirano et al., “Studies on the Absorption of Practically
`patent is extended or adjusted under 35
`Water—Insolable Drugs Following Injection VIII: Compari-
`US.C. 154(b) by 0 days.
`son of the Subcutaneous Absorption Rates From Aqueous
`Suspensions in the Mouse, Rat and Rabbit,” J. Pharm. Sci.,
`1993, 72(6), pp. 608-612.
`Nara et al. “A New Method for Assessment of Drug
`Absorption From Muscle: Application of a Local Perfusion
`System,” J. Pharm. Pharmacol, 1991, 43, pp. 272-274.
`Dickert et al., “Absorption of NPH-Insulin from Subcuta-
`neous Tissue: A Methodological Study in Pigs,” ACTA
`Pharmacol et Toxicol, 1982, 51, pp. 30-37.
`Supersaxo et al., “Effects of Molecular Weight on the
`Lymphatic Absorption of Water-Soluble Compounds Fol-
`lowing
`Subcutanous Administration,”
`Pharmaceutical
`Research, 1990, 7(2), pp. 167-169.
`Mosjov,“Structural Requirements for Biological Activity of
`Glucagon—Like Peptide-1, ” Int. J. Peptide Protein Res.,
`1992, 40, pp. 333-343.
`Suzuki et al., “Comparison of the Effects of Various C—Ter-
`minal and N-Terminal Fragment Peptide of Glucogen—Like
`Peptide-1 or Insulin and Glucagon Release from the Isolated
`Perfused Rat Pancreas,” Endocrinology, 1989, 125(6), pp.
`3109-3113.
`
`4,654,324 *
`3/1987 Chance et al.eee 514/12
`4,857,505 *
`8/1989 Ardent
`scecccccsssccssssssssssseesseeeeeee 514/2
`4,985,404 *
`1/1991 Mitchell
`...cccccccsssssssseseseseeseeee 514/6
`5,118,666 *
`6/1992 Habener......
`. 514/12
`
`5,120,712 *
`. 514/12
`6/1992 Habener ..
`5,175,145 * 12/1992 Cooper....
`. 514/4
`
`6/1995 EMG veeccccssscssssscssssssssssnseessesesseee 514/2
`5,424,286
`5,545,618
`8/1996 Buckley et al. wow $14/2
`11/1996 Johnsonet al.
`.
`5,574,008
`. 514/12
`
`3/1997 Habener weccssssessssssseeccsesseesseeee 514/12
`5,614,492
`5/1997 Efendic ...cscessssssssssessseseeseeee 514/12
`5,631,224
`
`Kréwezyfski, Leszek, “Technologia postaci lekéw (Tech-
`nology
`of
`the
`drug
`forms)’,
`Pafistwowy Zaklad
`Wydawnictw Lekarskich Warszawa,
`1969, pp.
`220,
`463-464, 490, 502.
`Gennaro, Alfonso R., Chapter 23 in Remington’s Pharma-
`ceutical Sciences, 16th edition, A. Osol Editor, Mack Pub-
`lishing Company, Easton, PA, 1980, pp. 343-363.
`Gill, I. J. et al., “Cyclodextrins as protection agents against
`enhancer damage in nasal delivery systems II. Effect on in
`2/1997 (DE).
`19530865 Al
`vivo absorption of insulin and histopathology of nasal
`4/1986 (EP).
`0177478
`membrane,” European Journal of Pharmaceutical Sciences
`11/1989 (EP).
`0 343 696 A2
`1:237-248, Jun. 1994.
`8/1991 (EP).
`0 442 671 A2
`Hendrick, G.K. et al., “Glucagon-like Peptide-I{7-37)
`3/1992 (EP).
`0 473 268 A3
`2/1993 (EP).
`0 526 862 Al
`Suppresses Hyperglycemia in Rats,” Metabolism 42(1):1-6,
`Jan. 1993.
`0 619 322 A2=10/1994 (EP).
`0658 568A1
`6/1995 (EP).
`0 733 644 Al
`9/1996 (EP).
`2 732 894
`10/1996 (FR).
`06941 * 11/1987 (WO) wees CO7K/7/10
`WO 88/07366
`10/1988 (WO).
`WO 90/02835
`3/1990 (WO).
`11296 * 10/1990 (WO) wees CO7K/7/10
`11457 *
`8/1991 (WO) wives CO7K/7/34
`WO 93/18785
`9/1993 (WO)
`WO 93/18786
`9/1993 (WO)
`WO 93/19175
`9/1993 (WO)
`WO 93/25579
`12/1993 (WO)
`WO 95/05848
`3/1995 (WO)
`WO 95/07098
`3/1995 (WO)
`WO 95/17510
`6/1995 (WO)
`WO 95/31214
`11/1995 (WO)
`WO 96/20005
`4/1996 (WO)
`
`FOREIGN PATENT DOCUMENTS
`
`(List continued on next page.)
`
`Primary Examiner—Sheela Huff
`(74) Attorney, Agent, or Firm—Sterne, Kessler, Goldstein
`& Fox P.L.L.C.
`
`ABSTRACT
`(57)
`There are disclosed methodsforthe treatment of non-insulin
`
`dependent diabetes mellitus in a mammal comprising the
`prolonged administration of GLP-1 (7-37), and related
`peptides. Also disclosed are compositions to prolong the
`administration of the peptides.
`
`37 Claims, 10 Drawing Sheets
`
`MPI EXHIBIT 1053 PAGE 1
`
`MPI EXHIBIT 1053 PAGE 1
`
`MPI EXHIBIT 1053 PAGE 1
`
`

`

`US 6,284,727 B1
`Page 2
`
`OTHER PUBLICATIONS
`
`Holst, J.J. et al., “Truncated glucago-like peptide I, an
`insulin—releasing hormonefor the distal gut,” FEBS Letters
`211(2):169-174, Jan. 1987.
`Remington’ Pharmaceutical Sciences, 16th ed. pp. 1555,
`1467, 1453, 1451, 1450, 1448, Ch. 91, 1980.*
`Crosset al., “Subcutaneous Absorption Kinetics and Local
`Tissue Distribution of Interferon and other Soluter,” J.
`Pharm. Pharmacol., 1993, 45, pp. 606-609;.
`Hirano et al., “Studies on the Absorption of Practically
`Water—-Insoluble Drugs following Injection VHI: Compari-
`son of the Subcutaneous Absorption Rates from Aqueous
`Suspensions in the Mouse, Rat, and Rabbit,” J. Pharm. Sci.,
`1993, 72(6), pp. 608-612;.
`Nara et al, “A New Method for Assessment of Drug
`Absorption from Muscle: Application of a Local Perfusion
`System,” J. Pharm. Pharmacol, 1991, 43, pp. 272-274;.
`
`Dickert et al., “Absorption of NPH-Insulin from Subcuta-
`neous Tissue: A Methodological Study in Pigs,” Acta Phar-
`macol et Toxicol, 1982, 51, pp. 30-37.
`Supersaxo et al., “Effects of Molecular Weight on the
`Lymphatic Absorption of Water-Soluble Compoundsfol-
`lowing Subcutaneoud Administration,” Pharmaceutical
`Research, 1990, 7(2), pp. 167-169;.
`Mosjov, “Structural requiremtns for Biological Activity of
`Glucagon-like Peptide-1,” Int. J. Peptide Protein Res.,
`1992, 40, pp. 333-343;.
`Suzuki et al., “Comparison of the Effects of various C—Ter-
`minal and N-Terminal Fragment Peptide of Glucogen-like
`Peptide-1 or Insulin and Glucagon Release from the Isolated
`Perfused Rat Pancreas,” endocrinology, 1989, 125(6), pp.
`3109-3113.
`
`* cited by examiner
`
`MPI EXHIBIT 1053 PAGE 2
`
`MPI EXHIBIT 1053 PAGE 2
`
`MPI EXHIBIT 1053 PAGE 2
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 1 of 10
`
`US 6,284,727 B1
`
`
`
`joUODOpabuojold‘€Gz'S6-dO@(9=U)
`
`
`
`(g=u)uolsnju|
`
`OeyO9€O0€OreO8tOclO09009-Ocl-
`
`UOISNJU]-UldosjoUI|NsUy
`
`08¢
`
`09¢
`
`Ove
`
`0d¢
`
`00
`N
`
`O8t
`
`09}
`
`OV
`
`Oct
`
`001
`
`BUSI
`
`esoon|y
`
`(jp/6w)
`
`MPI EXHIBIT 1053 PAGE 3
`
`MPI EXHIBIT 1053 PAGE 3
`
`MPI EXHIBIT 1053 PAGE 3
`
`
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 2 of 10
`
`US 6,284,727 B1
`
`(ujw)ew
`
`
`
`
` (g=u)Ju]owNfee-‘EGZ‘S6-dOe021
`(g=u)jonuogog«=U]HIpeulqu|
`OceO9€O0&OveOB8lOctO09009-Odl-
`
`08¢
`
`09¢
`
`Ove
`
`Ode
`
`00¢
`
`08}
`
`O91
`
`Ov
`
`OO}
`
`asoonj5
`
`BWSP|d
`
`(1p/6u)
`
`MPI EXHIBIT 1053 PAGE 4
`
`MPI EXHIBIT 1053 PAGE 4
`
`MPI EXHIBIT 1053 PAGE 4
`
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 3 of 10
`
`US 6,284,727 B1
`
`(g=u)urw/By/6ug‘eg2‘G6-dd@(g=u)
`joNU0DgO
`
`
`
`(g=u)utw/6»/6uy‘ege‘96-dOVW
`
`
`
`UOISNJU]UIdOJOUI;NSU|
`
`OcvO9€O0&OveO8tOclO09
`
`0 0
`
`9-
`Od1-
`
`081
`
`O91
`
`OVl
`
`Oct
`
`OOt
`
`MPI EXHIBIT 1053 PAGE 5
`
`€‘Sls
`
`08¢
`
`09¢
`
`Ove
`
`0c¢
`
`00¢
`(1p/6w)
`
`asoon|s)
`
`BWSP|d
`
`MPI EXHIBIT 1053 PAGE 5
`
`MPI EXHIBIT 1053 PAGE 5
`
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 4 of 10
`
`US 6,284,727 B1
`
`(hrs)
`
`Time
`
`(}w/6u)
`ewSeid Ul SUOIeJJUBDUOD UIdoJOUINsU|
`
`FIG.4
`
`MPI EXHIBIT 1053 PAGE 6
`
`MPI EXHIBIT 1053 PAGE 6
`
`MPI EXHIBIT 1053 PAGE 6
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 5 of 10
`
`US 6,284,727 B1
`
`(hrs)
`
`FIG.5
`
`Time
`
`10
`
`1
`
`m=
`
`(jw/6u)
`BWSE|q U] SUOI}E1JUGOUOD UldosjOUNNSUy
`
`.01
`
`MPI EXHIBIT 1053 PAGE 7
`
`MPI EXHIBIT 1053 PAGE 7
`
`MPI EXHIBIT 1053 PAGE 7
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 6 of 10
`
`US 6,284,727 B1
`
`NO
`NA
`ann
`
`<< --AS24—-f#--—AS25
`
`(hrs)
`
`FIG.6
`
`o
`
`7
`
`=
`
`Time
`
`01
`
`(1w/6u)
`BUSB|q U] SUOHEIUSOUOD UldojOUI|Nsu|
`
`MPI EXHIBIT 1053 PAGE 8
`
`MPI EXHIBIT 1053 PAGE 8
`
`MPI EXHIBIT 1053 PAGE 8
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 7 of 10
`
`US 6,284,727 B1
`
`o-
`
`AS31
`
`©)
`
`N w<
`
`x
`
`FIG.7
`
`(hrs)
`
`Time
`
`oS
`=
`
`”
`
`~-
`.
`
`(jw1/6u)
`BWSEPjq Uj SUO!}B1]JUBDUOD UIdOJ}OUL|NSU|
`
`01
`
`MPI EXHIBIT 1053 PAGE 9
`
`MPI EXHIBIT 1053 PAGE 9
`
`MPI EXHIBIT 1053 PAGE 9
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 8 of 10
`
`US 6,284,727 B1
`
`FIG. 8
`
`1000
`
`100
`
`—_ oO
`
`
`
`
`
`Insulinotropin(ng/miplasma)
`
`0.1
`
`0
`
`5
`
`10
`
`15
`Time (hours)
`
`20
`
`25
`
`30
`
`MPI EXHIBIT 1053 PAGE 10
`
`MPI EXHIBIT 1053 PAGE 10
`
`MPI EXHIBIT 1053 PAGE 10
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 9 of 10
`
`US 6,284,727 B1
`
`FIG. 9
`
`100
`
`
`
`
`
`Insulinotropin(ng/mlplasma)
`
`10
`
` 0
`
`25
`
`30
`
`MPI EXHIBIT 1053 PAGE11
`
`5
`
`10
`
`15
`Time (hours)
`
`20
`
`MPI EXHIBIT 1053 PAGE 11
`
`MPI EXHIBIT 1053 PAGE 11
`
`

`

`U.S. Patent
`
`Sep. 4, 2001
`
`Sheet 10 of 10
`
`US 6,284,727 B1
`
`~~,
`”
`£
`@
`E
`
`a
`
`Ce)
`QI
`
`oO
`ql
`
`in
`bo
`
`©
`
`lO
`
`Oo
`
`~
`S
`
`wn
`3
`/2
`oO.
`:
`<
`:
`
`=>
`O
`
`®
`£
`ra}
`2
`oO
`
`||
`
`a
`
`©
`2
`
`Oo
`oe
`
`rr
`
`2o
`
`D
`OL
`
`MPI EXHIBIT 1053 PAGE 12
`
`©
`
`eel
`
`a©
`
`a L
`
`L.
`
`MPI EXHIBIT 1053 PAGE 12
`
`MPI EXHIBIT 1053 PAGE 12
`
`

`

`US 6,284,727 B1
`
`1
`PROLONGED DELIVERY OF PEPTIDES
`
`This application is a continuation of application Ser. No.
`08/181,655, filed Jan. 25, 1994, now abandoned, which is a
`continuation-in-part of application Ser. No. 08/044,133,
`filed Apr. 7, 1993, now abandoned.
`BACKGROUND OF THE INVENTION
`
`The present invention relates to compositions and meth-
`ods for the treatment of Diabetes Mellitus. Morespecifically,
`the present invention relates to compositions to prolong the
`administration of glucagon-like peptide 1 (GLP-1), and
`derivatives thereof. These compositions are useful in treat-
`ment of Non-Insulin Dependent Diabetes Mellitus
`(NIDDM).
`The amino acid sequence of GLP-1 is knownas:
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1)
`GLP-1 is disclosed by Lopez, L. C., et al., PN.A.S., USA
`80: 5485-5489 (1983); Bell, G. L, et al., Nature 302:
`716-718 (1983); Heinrich, G. et al, Endocrinol. 115:
`2176-2181 (1984) and Ghiglione, M., et al., Diabetologia
`27: 599-600 (1984).
`During processing in the pancreas and intestine, GLP-1 is
`converted to a 31 amino acid peptide having amino acids
`7-37 of GLP-1, hereinafter this peptide is referred to as
`GLP-1 (7-37).
`This peptide has been shown to have insulinotropic
`activity,
`that
`is,
`it
`is able to stimulate, or cause to be
`stimulated,
`the synthesis or expression of the hormone
`insulin. Because of this insulinotropic activity, GLP-1
`(7-37) is alternatively referred to as insulinotropin.
`GLP-1 (7-37) has the following amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCE ID NO: 2).
`GLP-1 (7-37), certain derivatives thereof and the use
`thereof to treat Diabetes mellitus in a mammalare disclosed
`in US. Pat. Nos. 5,118,666 (666 patent) and 5,120,712
`(712 patent), the disclosures of these patents being incor-
`porated herein by reference. The derivatives of GLP-1
`(7-37) disclosed in the ’666 and ’712 patents include
`polypeptides which contain or lack one of more amino acids
`that may not be present in the naturally occurring sequence.
`Further derivatives of GLP-1 (7-37) disclosed in the *666
`and *712 patents include certain C-terminal salts, esters and
`amides where the salts and esters are defined as OM where
`M is a pharmaceutically acceptable cation or a lower
`(C,—-C,) branched or unbranched alkyl group and the amides
`are defined as —NR*R* where R* and R® are the same or
`different and are selected from the group consisting of
`hydrogen and a lower (C,—C,) branched or unbranchedalkyl
`group.
`Certain other polypeptides, alternatively referred to as
`truncated GLP-1 or truncated insulinotropin, having insuli-
`notropic activity and the derivatives thereof are disclosed in
`PCT/US 89/01121 (WO 90/11296). Those polypeptides,
`referred to therein as GLP-1 (7-36), GLP-1 (7-35) and
`GLP-1 (7-34) have the following amino acid sequences,
`respectively.
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg (SEQUENCEID NO: 3);
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly (SEQUENCEID NO: 4); and
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`2
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys (SEQUENCE ID NO: 5);
`Derivatives of the polypeptides disclosed in PCT/US89/
`01121 include polypeptides having inconsequential amino
`acid substitutions, or additional amino acids to enhance
`coupling to carrier protein or to enhance the insulinotropic
`effect thereof. Further derivatives of insulinotropin disclosed
`in PCT/US89/01121 include certain C-terminal salts, esters
`and amides where the salts and esters are defined as OM
`
`where M is a pharmaceutically acceptable cation or a lower
`branched or unbranched alkyl group and the amides are
`defined as —NR*R* where R* and R® are the same or
`different and are selected from the group consisting of
`hydrogen and a lower branched or unbranched alkyl group.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`FIG. 1 showsthe effect of a prolonged infusion (7 hours)
`of 4 ng/kg/min insulinotropin on plasma glucose levels in
`patients with NIDDM.
`FIG. 2 showsthe effect of a short infusion (60 minutes)
`of 10 ng/kg/min insulinotropin on plasma glucose levels in
`patients with NIDDM.
`FIG. 3 showsthe effect of a prolonged infusion (7 hours)
`of 2 ng/kg/min and 4 ng/kg/min of insulinotropin on plasma
`glucose levels in patients with NIDDM.
`FIG. 4. Mean (n=3) Plasma Concentration of Insulinotro-
`pin in Rats After Subcutaneous Administration of Single 0.5
`mg/0.5 ml Doses in Different Aqueous Suspensions (AS).
`FIG. 5. Mean (n=3) Plasma Concentration of Insulinotro-
`pin in Rats After Subcutaneous Administration of Single 0.5
`mg/0.5 ml Doses in Different Aqueous Suspensions (AS).
`FIG. 6. Mean (n=3) Plasma Concentration of Insulinotro-
`pin in Rats After Subcutaneous Administration of Single 0.5
`mg/0.5 ml Doses in Different Aqueous Suspensions (AS).
`FIG. 7. Mean (n=3) Plasma Concentration of Insulinotro-
`pin in Rats After Subcutaneous Administration of Single 0.5
`mg/0.5 ml Doses in Different Aqueous Suspensions (AS).
`FIG. 8. Mean (n=3) Plasma Concentration of Insulinotro-
`pin in Rats After Subcutaneous Administration of Single 0.5
`mg/0.13 ml Doses in Different Aqueous Suspensions (AS).
`FIG. 9. Mean (n=3) Plasma Concentration of Insulinotro-
`pin in Rats After Subcutaneous Administration of Single 0.5
`mg/0.13 ml Doses in Different Aqueous Suspensions (AS).
`FIG. 10 shows pharmacokinetic studies of an insulinotro-
`pin zinc precipitate.
`
`
`
`SUMMARY OF THE INVENTION
`
`In one embodiment, the present invention is directed to a
`method for the treatment of non-insulin dependent diabetes
`mellitus in a mammalin need of such treatment comprising
`the repeated administration over an extended period of time
`of a compound with prolonged action after each
`administration, said prolonged action necessary to achieve
`sustained glycemic control in such mammals, said com-
`pound selected from the group consisting of:
`(a) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly; (SEQUENCE ID NO: 2)
`(b) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-X (SEQUENCEID NO: 7)
`
`MPI EXHIBIT 1053 PAGE 13
`
`MPI EXHIBIT 1053 PAGE 13
`
`MPI EXHIBIT 1053 PAGE 13
`
`

`

`US 6,284,727 B1
`
`3
`wherein X is selected from the group consisting of:
`(A) Lys,
`(B) Lys-Gly, and
`(C) Lys-Gly-Arg;
`(c) a derivative of a polypeptide comprising the primary
`structure
`
`5
`
`H,N—W—COOH
`
`wherein W is an amino acid sequence selected from the
`group consisting of
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1) and
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg
`(SEQUENCEID NO: 6)
`which derivative when processed in a mammalresults in a
`polypeptide derivative having an insulinotropic activity;
`(d) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—R—COOH
`
`wherein R is an amino acid sequence selected from the
`group consisting of
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly; (SEQUENCE ID NO: 2)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg; (SEQUENCE ID NO: 3)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly; (SEQUENCE ID NO: 4) and
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys; (SEQUENCEID NO: 5) and
`(e) a derivative of said peptides (a) through (d) wherein
`said derivative is selected from the group consisting of:
`(1) a pharmaceutically acceptable acid addition salt of
`said peptides;
`(2) a pharmaceutically acceptable carboxylate salt of said
`peptides;
`(3) a pharmaceutically acceptable alkali addition salt of
`said peptides;
`(4) a pharmaceutically acceptable lower alkyl ester of said
`peptides; and
`(5) a pharmaceutically acceptable amide of said peptides
`wherein said pharmaceutically acceptable amide is
`selected from the group consisting of amide,
`lower
`alkyl amide and lower dialkyl amide.
`Preferred is the method wherein said administration is
`subcutaneous.
`Also preferred is the method wherein said administration
`is intramuscular.
`
`Also preferred is the method wherein said administration
`is transdermal.
`
`Also especially preferred is the method wherein said
`administration is by an infusion pump.
`Also preferred is the method wherein said administration
`is by oral inhalation.
`Also preferred is the method wherein said administration
`is by nasal inhalation.
`Also preferred is the method wherein said administration
`is gastrointestinal.
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`4
`In another embodiment, the present invention is directed
`to a composition of matter comprising;
`(i) a compound selected from the group consisting of:
`(a) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly; (SEQUENCE ID NO: 2)
`(b) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-X (SEQUENCEID NO: 7)
`wherein X is selected from the group consisting of:
`(A) Lys,
`(B) Lys-Gly, and
`(C) Lys-Gly-Arg;
`(c) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—W—COOH
`
`wherein W is an amino acid sequence selected from the
`group consisting of
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu- Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1) and
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg
`(SEQUENCEID NO: 6)
`which derivative when processed in a mammalresults in a
`polypeptide derivative having an insulinotropic activity;
`(d) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—R—COOH
`
`wherein R is an amino acid sequence selected from the
`group consisting of
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCEID NO: 2)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg (SEQUENCEID NO: 3)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly (SEQUENCEID NO: 4) and
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys (SEQUENCE ID NO: 5);
`a derivative of said peptides (a) through (d) wherein said
`derivative is selected from the group consisting of:
`(1) a pharmaceutically acceptable acid addition salt of
`said peptides;
`(2) a pharmaceutically acceptable carboxylate salt of said
`peptides;
`(3) a pharmaceutically acceptable alkali addition salt of
`said peptides;
`(4) a pharmaceutically acceptable lower alkyl ester of said
`peptides; and
`(5) a pharmaceutically acceptable amide of said peptides
`wherein said pharmaceutically acceptable amide is
`selected from the group consisting of amide,
`lower
`alkyl amide and lower dialkyl amide, and
`(ii) a polymer capable of prolonging the action of said
`compound to achieve sustained glycemic control.
`
`MPI EXHIBIT 1053 PAGE 14
`
`MPI EXHIBIT 1053 PAGE 14
`
`MPI EXHIBIT 1053 PAGE 14
`
`

`

`US 6,284,727 B1
`
`5
`Especially preferred is the composition wherein said
`polymer is a low molecular weight polymer.
`Further especially preferred is a composition wherein said
`polymer is selected from the group consisting of polyeth-
`ylene glycol, polyvinylpyrrolidone, polyvinylalcohol,
`polyoxyethylene-polyoxypropylene copolymers, polysac-
`charides selected from the group consisting of cellulose,
`cellulose derivatives, chitosan, acacia gum, karaya gum,
`guar gum, xanthan gum,
`tragacanth, alginic acid,
`carrageenan, agarose, and furcellarans, dextran, starch,
`starch derivatives, hyaluronic acid, polyesters, polyamides,
`polyanhydrides, and polyortho esters, with especially pre-
`ferred polymers selected from the group consisting of poly-
`ethylene glycol and polyvinylpyrrolidone.
`In another embodiment, the present invention is directed
`to a composition of matter comprising;
`(i) a compound selected from the group consisting of:
`(a) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCE ID NO: 2);
`(b) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-X (SEQUENCE ID NO: 7)
`wherein X is selected from the group consisting of:
`(A) Lys,
`(B) Lys-Gly, and
`(C) Lys-Gly-Arg;
`(c) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—W—COOH
`
`wherein W is an amino acid sequence selected from the
`group consisting of
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1) and
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg
`(SEQUENCEID NO: 6)
`which derivative when processed in a mammalresults in a
`polypeptide derivative having an insulinotropic activity;
`(d) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—R—COOH
`
`wherein R is an amino acid sequence selected from the
`group consisting of
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCE ID NO:2);
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg (SEQUENCEID NO: 3);
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly (SEQUENCEID NO: 4); and
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys (SEQUENCEID NO: 5); and
`a derivative of said peptides (a) through (d) wherein said
`derivative is selected from the group consisting of:
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`6
`(1) a pharmaceutically acceptable acid addition salt of
`said peptides;
`(2) a pharmaceutically acceptable carboxylate salt of said
`peptides;
`(3) a pharmaceutically acceptable alkali addition salt of
`said peptides;
`(4) a pharmaceutically acceptable lower alkyl ester of said
`peptides; and
`(5) a pharmaceutically acceptable amide of said peptides
`wherein said pharmaceutically acceptable amide is
`selected from the group consisting of amide,
`lower
`alkyl amide and lower dialkyl amide, and
`(ii) a pharmaceutically acceptable water-immiscible oil
`suspension capable of prolonging administration of
`said compound.
`Especially preferred is the composition wherein said oil is
`selected from the group consisting of peanutoil, sesameoil,
`almondoil, castor oil, camellia oil, cotton seed oil, olive oil,
`corn oil, soy oil, safflower oil, coconut oil, esters of fatty
`acids, and esters of fatty alcohols.
`Further especially preferred is the composition further
`comprising a wetting agent, especially a nonionic surfactant.
`More further especially preferred is the composition fur-
`ther comprising a suspending agent.
`In another embodiment, the present invention is directed
`to a composition of matter comprising;
`(i) a compound selected from the group consisting of:
`(a) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCEID NO: 2);
`(b) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-X (SEQUENCEID NO: 7)
`wherein X is selected from the group consisting of:
`(A) Lys,
`(B) Lys-Gly, and
`(C) Lys-Gly-Arg;
`(c) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—W—COOH
`
`wherein W is an amino acid sequence selected from the
`group consisting of
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu- Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1) and
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg
`(SEQUENCEID NO: 6)
`which derivative when processed in a mammalresults in a
`polypeptide derivative having an insulinotropic activity;
`(d) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—R—COOH
`
`wherein R is an amino acid sequence selected from the
`group consisting of
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCEID NO: 2)
`
`MPI EXHIBIT 1053 PAGE 15
`
`MPI EXHIBIT 1053 PAGE 15
`
`MPI EXHIBIT 1053 PAGE 15
`
`

`

`US 6,284,727 B1
`
`7
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg (SEQUENCEID NO: 3)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly (SEQUENCEID NO: 4) and
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys (SEQUENCEID NO: 5); and
`a derivative of said peptides (a) through (d) wherein said
`derivative is selected from the group consisting of:
`(1) a pharmaceutically acceptable acid addition salt of
`said peptides;
`(2) a pharmaceutically acceptable carboxylate salt of said
`peptides;
`(3) a pharmaceutically acceptable alkali addition salt of
`said peptides;
`(4) a pharmaceutically acceptable lower alkyl ester of said
`peptides; and
`(5) a pharmaceutically acceptable amide of said peptides
`wherein said pharmaceutically acceptable amide is
`selected from the group consisting of amide,
`lower
`alkyl amide and lower dialkyl amide, and
`(ii) zine (II), which is complexed with the peptide.
`Preferred is the composition capable of sustained glyce-
`mic action.
`Especially preferred is the composition wherein the zinc
`product is amorphous.
`Also especially preferred is the composition wherein the
`zine productis crystalline.
`invention is
`the present
`In yet another embodiment,
`directed to a composition of matter comprising;
`(i) a compound selected from the group consisting of:
`(a) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCE ID NO: 2);
`(b) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gin-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-X (SEQUENCE ID NO: 7)
`wherein X is selected from the group consisting of:
`(A) Lys,
`(B) Lys-Gly, and
`(C) Lys-Gly-Arg;
`(c) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—W—COOH
`
`wherein W is an amino acid sequence selected from the
`group consisting of
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1) and
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg
`(SEQUENCEID NO: 6)
`and which derivative when processed in a mammalresults in
`a polypeptide derivative having an insulinotropic activity;
`(d) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—R—COOH
`
`10
`
`15
`
`20
`
`25
`
`30
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`8
`wherein R is an amino acid sequence selected from the
`group consisting of
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCEID NO: 2)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg (SEQUENCEID NO: 3)
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly (SEQUENCEID NO: 4) and
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys (SEQUENCEID NO: 5); and
`a derivative of said peptides (a) through (d) wherein said
`derivative is selected from the group consisting of:
`(1) a pharmaceutically acceptable acid addition salt of
`said peptides;
`(2) a pharmaceutically acceptable carboxylate salt of said
`peptides;
`(3) a pharmaceutically acceptable alkali addition salt of
`said peptides;
`(4) a pharmaceutically acceptable lower alkyl ester of said
`peptides; and
`(5) a pharmaceutically acceptable amide of said peptides
`wherein said pharmaceutically acceptable amide is
`selected from the group consisting of amide,
`lower
`alkyl amide and lower dialkyl amide, and
`(ii) a metal selected from the group consisting of Ni (ID,
`Co GD, Mg (ID, Ca (ID, K (1), Mn dI), Fe(ID, and
`CuI).
`invention is
`the present
`In yet another embodiment,
`directed to a composition of matter comprising;
`(i) a compound selected from the group consisting of:
`(a) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCEID NO: 2);
`(b) a peptide having the amino acid sequence:
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-X (SEQUENCEID NO: 7)
`wherein X is selected from the group consisting of:
`(A) Lys,
`(B) Lys-Gly-,
`(C) Lys-Gly-Arg;
`(c) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—W—COOH
`
`wherein W is an amino acid sequence selected from the
`group consisting of
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu- Val-Lys-Gly-Arg-Gly
`(SEQUENCEID NO: 1) and
`His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-
`Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-
`Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg
`(SEQUENCEID NO: 6)
`which derivative when processed in a mammalresults in a
`polypeptide derivative having an insulinotropic activity;
`
`MPI EXHIBIT 1053 PAGE 16
`
`MPI EXHIBIT 1053 PAGE 16
`
`MPI EXHIBIT 1053 PAGE 16
`
`

`

`US 6,284,727 B1
`
`9
`(d) a derivative of a polypeptide comprising the primary
`structure
`
`H,N—R—COOH
`
`wherein R is an amino acid sequence selected from the
`group consisting of
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg-Gly (SEQUENCE ID NO: 2);
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp- Val-Ser-Ser-Tyr-
`Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-
`Leu-Val-Lys-Gly-Arg (SEQUEN