I 1111111111111111 1111111111 111111111111111 IIIII IIIII IIIII IIIIII IIII IIII IIII
`US010335462B2
`
`c12) United States Patent
`Jensen
`
`(IO) Patent No.: US 10,335,462 B2
`(45) Date of Patent:
`*Jul. 2, 2019
`
`(54) USE OF LONG-ACTING GLP-1 PEPTIDES
`
`(71) Applicant: Novo Nordisk A/S, Bagsvaerd (DK)
`
`(72)
`
`Inventor: Christine Bjoern Jensen,
`Charlottenlund (DK)
`
`(73) Assignee: Novo Nordisk A/S, Bagsvaerd (DK)
`
`( *) Notice:
`
`Subject to any disclaimer, the term ofthis
`patent is extended or adjusted under 35
`U.S.C. 154(b) by O days.
`
`This patent is subject to a terminal dis(cid:173)
`claimer.
`
`(21) Appl. No.: 15/656,042
`
`(22) Filed:
`
`Jul. 21, 2017
`
`(65)
`
`Prior Publication Data
`
`US 2018/0085435 Al Mar. 29, 2018
`
`Related U.S. Application Data
`
`(63) Continuation of application No. 14/409,493, filed as
`application No. PCT/EP2013/063004 on Jun. 21,
`2013, now Pat. No. 9,764,003.
`
`(60) Provisional application No. 61/708,162, filed on Oct.
`1, 2012, provisional application No. 61/694,837, filed
`on Aug. 30, 2012.
`
`(30)
`
`Foreign Application Priority Data
`
`Jul. 1, 2012
`Oct. 1, 2012
`
`(EP)
`(EP)
`
`12174535
`12186781
`
`(51)
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`Int. Cl.
`A61K 38/26
`A61P5/50
`A61P3/04
`A61P3/10
`(52) U.S. Cl.
`CPC .................................... A61K 38/26 (2013.01)
`( 58) Field of Classification Search
`None
`See application file for complete search history.
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`8,129,343 B2
`8,536,122 B2
`2010/0047762 Al
`2010/0292133 Al
`2011/0301080 Al
`
`3/2012 Lau et al.
`9/2013 Lau et al.
`2/2010 Button et al.
`11/2010 Spetzler et al.
`12/2011 Bush et al.
`
`FOREIGN PATENT DOCUMENTS
`
`CN
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`
`102229668 A
`2409349 C2
`
`11/2011
`1/2011
`
`RU
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`
`2413530 C2
`2006/097537 A2
`2010/092163 A2
`2011/080103 Al
`2011138421 Al
`12016419 Al
`2012080471 Al
`2012107476 Al
`12130136 Al
`12136790 Al
`2012177929 A2
`
`3/2011
`9/2006
`8/2010
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`8/2012
`10/2012
`10/2012
`12/2012
`
`OTHER PUBLICATIONS
`
`Madsbad S et al. An Overview of once-weekly glucagon-like
`peptide-1 receptor agonists available efficacy and safety data and
`perspectives for the future, Diabetes, Obesity and Metabolism Year
`2011, vol. 13, No. 5, pp. 394-407.
`Buse B. J. et al., Exenatide once weekly versus liraglutide once
`daily in patients with type 2 diabetes (DURATION-6): a randomised,
`open-label study, Lancet, 2013, vol. 381, pp. 117-124.
`Kim et al. "Effects of Once-Weekly Dosing of a Long-Acting
`Release Formulation of Exentide on Glucose Control and Body
`Weight in Subjects with Type 2 Diabetes." Diabetes care (2007) vol.
`30 pp. 1487-1493.
`Bydureon NDA 022200/S-008 Package Information pp. 1-179 (Feb.
`2014).
`Clinical Trial NCT00696657 entitled "A Randomized Controlled
`Clinical Trial in Type 2 Diabetes Comparing Semaglutide to Pla(cid:173)
`cebo and Liraglutide." pp. 1-5 Mar. 2015. Accessed Sep. 24, 2015
`at clinicaltrials.gov/archive/NCT00696657/2011_03 _25.
`Lau et al. "Discovery of the Once-WEekly Glucagon-Like Peptide-1
`(GLP-1) Analogue Semaglutide." J. Med. Chem. (2015) vol. 58 pp.
`7370-7380.
`Eperzan Assessment Report. Euro. Med. Agency. pp. 1-124 (2014)
`accessed Sep. 24, 2015 at URL ema.europa.ed/docs/en_ GB/document_
`Ii brary/EPAR_ -_Public _assessment_report/human/002 73 5/
`WC500165119.pdf.
`Trulicity Assessment Report. Euro. Med. Agency. pp. 1-172 (2014)
`accessed Sep. 24, 2015 at URL ema.europa.ed/docs/en_ GB/document_
`Ii brary/EPAR_ -_Public _assessment_report/human/002 82 5/
`WC500179473.pdf.
`Mizuta et al. "The Role for GLP-1 in Regulation of Body Weight."
`Progress in Medicine 2008 vol. 28 No. 8 pp. 1909-1912.
`CDC, "National Health and Nutrition Examination Survey: Healthy
`Weight, Overweight and Obesity among U.S. adults" 03-0260 pp.
`1-2 (Jul. 2003), accessed May 10, 2016 at URL cdc.gov/nchs/data/
`nhanes/databriefs/adultweight.pdf.
`
`Julie Ha
`Primary Examiner -
`Assistant Examiner - Kristina M Hellman
`(74) Attorney, Agent, or Firm - Leon Y. Lum
`
`(57)
`
`ABSTRACT
`
`The invention relates to use of long-acting GLP-1 peptides
`in certain dosage regimes for the treatment of type 2
`diabetes, obesity, etc.
`
`10 Claims, 6 Drawing Sheets
`Specification includes a Sequence Listing.
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00001
`
`

`

`U.S. Patent
`
`Jul. 2, 2019
`
`Sheet 1 of 6
`
`US 10,335,462 B2
`
`Semaglutide
`Liraglutide
`8.0
`8.1
`8.1
`8.1
`8.1
`8.2
`8.2
`8.2
`Baseline 8.1
`Placebo 0, 1 mg 0.2 rng 0.4 mg 0,8 mg 0.8 mg ·1 .6 mg 1,2 mg 1.8 rng
`T
`T
`o.oo---
`
`-0.2S 0i
`
`0 -0.50
`~
`--;-0.75
`:a I
`
`-1.00
`
`-1.25
`
`-1.50
`
`-1.75
`
`-2.00
`
`-1.18
`
`-1.34
`
`-1.69
`
`*p<0.05 vs. placebo
`·•*p<0.001 vs. placebo
`11Semaglutide 1.6 mg T superior to Hrag!utide 1.2 mg and ·1 ,8 mg
`Data are LS means,
`
`Fig. 1
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00002
`
`

`

`N = N
`
`0--,
`~
`UI
`
`"'""' = w w
`d r.,;_
`
`0 ....
`N
`.....
`rJJ =(cid:173)
`
`('D
`('D
`
`O'I
`
`0 ....
`N
`~
`:-'
`2'
`
`1,0
`
`~ = ~
`
`~
`~
`~
`
`00 .
`e .
`
`·----~-------.. .. . ~.._::7.._--. ,t:4a . ----i
`
`12
`
`11
`
`10
`
`9
`
`8
`
`7
`
`6
`
`5
`
`4
`
`Fig. 2
`
`--+--!irag!utide 1,8 rn9
`-o-serrn~9!utide 0,8 rng T
`
`❖--· sernag!utide 0.2 mg
`
`-e-semag!utide L6 mg T ········*-····----!irag!utide L2. rng
`·····*·····Semagkrtide 0,4 mg ~smnag!ubde O,H rn9
`semag!utide 0.1 mg
`.w.·-t=:··-w placebo
`
`F
`
`r
`
`-2,0
`··L8
`-L6
`-1,4
`-L2
`-LO
`-0,8
`··0,6
`--OA
`-0.2
`0,0
`
`.c u
`ro
`C
`01
`(I)
`
`..-!
`<.,.)
`
`C
`J:
`..0
`<t:
`
`3
`
`2
`
`1
`
`0
`
`Time
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00003
`
`

`

`U.S. Patent
`
`Jul. 2, 2019
`
`Sheet 3 of 6
`
`US 10,335,462 B2
`
`<(
`C") .
`O') u::
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00004
`
`

`

`U.S. Patent
`
`Jul. 2, 2019
`
`Sheet 4 of 6
`
`US 10,335,462 B2
`
`,~ • .,~.;;
`· ii I;
`, , .. ,y··· ,-~- , .. ,r··· ,'{'··· «p··· y···· y···· 'i, >+ l ::;:
`
`:ml:
`
`·$'
`
`al
`
`cw, . en ·-LL
`
`j ···· .. ·
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00005
`
`

`

`N = N
`
`0--,
`~
`UI
`
`"'""' = w w
`
`Fig. 4
`
`rJl.
`d
`
`---+--lirag!utide LS mg
`-o-semag!utide (LS rng T
`..... -:-, .... semaglutide O ,2 mg
`
`-+-semaglutide 0.8 rng
`semaglutide 0, 1 mg
`
`,....._.sen-iag!utide L6 mg T ·········*··········Hraglutide L2 mg
`···-:-~}.,••···Semaglutide QA. rng
`·····$:::···· placebo
`
`0 ....
`Ul
`.....
`rJJ =(cid:173)
`
`('D
`('D
`
`O'I
`
`0 ....
`N
`~
`:-'
`2'
`
`1,0
`
`~ = ~
`
`~
`~
`~
`
`00 .
`e .
`
`-i;;;~E;;,·····=~~•~·.·.·.•·•·•·•·•······••~··~
`
`1
`
`-:-·
`
`12
`
`1.1 ,1,
`
`10
`
`9
`
`s
`
`7
`
`6
`
`5
`
`4
`
`3
`
`2
`
`1
`
`0
`
`Time (weeks)
`
`-6
`
`-5
`
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`
`-3
`
`N2
`
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`
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`
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`;,:;
`©
`rn
`,t;;._
`
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`
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`
`,::;i. -(cid:173) .....
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00006
`
`

`

`U.S. Patent
`
`Jul. 2, 2019
`
`Sheet 6 of 6
`
`US 10,335,462 B2
`
`U') .
`C, ·-u.
`
`- ~
`
`
`Q
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`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00007
`
`

`

`US 10,335,462 B2
`
`1
`USE OF LONG-ACTING GLP-1 PEPTIDES
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`2
`less often, and wherein said GLP-1 agonist and/or admin(cid:173)
`istration optionally is as defined herein.
`
`BRIEF DESCRIPTION OF DRAWINGS
`
`This application is a continuation of U.S. application Ser.
`No. 14/409,493, filed Dec. 19, 2014, which is a 35 U.S.C. §
`371 National Stage application oflntemational Application
`PCT/EP2013/063004 (WO 2014/005858), filed Jun. 21,
`2013, which claimed priority of European Patent Applica(cid:173)
`tion 12174535.0, filed Jul. 1, 2012 and European Patent
`Application 12186781.6, filed Oct. 1, 2012; this application
`claims priority under 35 U.S.C. § 119 of U.S. Provisional
`Application 61/694,837; filed Aug. 30, 2012 and U.S. Pro- 15
`visional Application 61/708,162; filed Oct. 1, 2012.
`The present invention relates to improved uses of GLP-1
`peptides in therapy.
`
`SEQUENCE LISTING
`
`The instant application contains a Sequence Listing which
`has been submitted in ASCII format via EFS-Web and is
`hereby incorporated by reference in its entirety. Said ASCII
`copy, created on Jun. 17, 2013 and amended on Jul. 12, 25
`2017, is named 8545US02_SeqList.txt and is 7,975 bytes in
`size.
`
`SUMMARY
`
`FIG. 1 shows change in HbAlc following subcutaneous
`administration of placebo, semaglutide, or liraglutide to
`human subjects. *p<0.05 vs. placebo; **p<0.001 vs. placebo
`(based on adjusted means). Baseline values are for infor-
`10 mation only: data are model-adjusted for baseline HbAlc.
`Data are model-adjusted LS means, FAS LOCF. The esti(cid:173)
`mates are from an ANOVA model with treatment, country
`and previous treatment as fixed effects and baseline HbAlc
`as covariate.
`FIG. 2 shows mean change in HbAlc from baseline
`versus time; data are mean (1.96SE), FAS LOCF. The
`treatments are placebo (A); semaglutide 0.1 mg (B, dashed
`line), 0.2 mg (C), 0.4 mg (D), 0.8 mg (E), 0.8 mg T (F,
`dashed line), 1.6 mg T (G); liraglutide 1.2 mg (H), 1.8 mg
`20 (I).
`FIG. 3A and FIG. 3B show subjects reaching the AACE
`(FIG. 3A) or ADA (FIG. 3B) criteria for glycaemic control.
`The number of patients reaching the criteria per treatment is
`indicated in each bar. The treatments are placebo (A);
`semaglutide 0.1 mg (B), 0.2 mg (C), 0.4 mg (D), 0.8 mg (E),
`0.8 mg T (F), 1.6 mg T (G); liraglutide 1.2 mg (H), 1.8 mg
`(I). *p<0.05 vs. placebo; **p<0.001 vs. placebo;
`***p<0.0001 vs. placebo (based on adjusted means). Data
`are FAS LOCF. The estimates are from a logistic regression
`30 model treatment, country and previous treatment as fixed
`effects and baseline HbAlc as covariate. ADA, American
`Diabetes Association; AACE, American Association of
`Clinical Endocrinologists.
`FIG. 4 shows mean body weight change versus time; data
`35 are mean (1.96SE), FAS LOCF. The treatments are placebo
`(A); semaglutide 0.1 mg (B, dashed line), 0.2 mg (C), 0.4 mg
`(D), 0.8 mg (E), 0.8 mg T (F, dashed line), 1.6 mg T (G);
`liraglutide 1.2 mg (H), 1.8 mg (I).
`FIG. 5 shows body weight change from baseline at week
`40 12. **p<0.001 vs. placebo; ***p<0.0001 vs. placebo (based
`on adjusted means. t: Baseline values for information only:
`data are model-adjusted for baseline weight. Data are model(cid:173)
`adjusted LS means, FAS LOCF. The estimates are from an
`ANOVA model with treatment, country and previous treat-
`45 ment as fixed effects and baseline weight as covariate.
`SE: Standard error. FAS: Full analysis set. LOCF: Last
`observation carried forward.
`
`In one embodiment the invention relates to a method for
`a) reduction of HbAlc; b) prevention or treatment of type 2
`diabetes, hyperglycemia, impaired glucose tolerance, or
`non-insulin dependent diabetes; or c) prevention or treat(cid:173)
`ment of obesity, reducing body weight and/or food intake, or
`inducing satiety; wherein said method comprises adminis(cid:173)
`tration of a GLP-1 agonist to a subject in need thereof,
`wherein said GLP-1 agonist i) has a half-life of at least 72
`hours, wherein said half-life optionally is determined by
`Assay (II); ii) is administered in an amount of at least 0.7 mg
`per week, such an amount equivalent to at least 0.7 mg
`semaglutide per week; and iii) is administered once weekly
`or less often.
`In one embodiment the invention relates to a GLP-1
`agonist for use in a) the reduction of HbAlc; b) the preven(cid:173)
`tion or treatment of type 2 diabetes, hyperglycemia,
`impaired glucose tolerance, or non-insulin dependent dia(cid:173)
`betes; or c) the prevention or treatment of obesity, for
`reducing body weight and/or food intake, or for inducing 50
`satiety; wherein said use comprises administration of said
`GLP-1 agonist in an amount of at least 0.7 mg per week,
`such an amount equivalent to at least 0. 7 mg semaglutide per
`week, and wherein said GLP-1 agonist and/or administration
`optionally is as defined herein.
`In one embodiment the invention relates to a composition
`comprising a GLP-1 agonist for use in a) the reduction of
`HbAlc; b) the prevention or treatment of type 2 diabetes,
`hyperglycemia, impaired glucose tolerance, or non-insulin
`dependent diabetes; or c) the prevention or treatment of 60
`obesity, for reducing body weight and/or food intake, or for
`inducing satiety; wherein said GLP-1 agonist i) has a
`half-life of at least 72 hours, wherein said half-life optionally
`is determined by Assay (II); and ii) is administered in an
`amount of at least 0.7 mg per week, such an amount 65
`equivalent to at least 0.7 mg semaglutide per week; and
`wherein said composition is administered once weekly or
`
`DESCRIPTION
`
`The present invention relates to an improved use of
`GLP-1 agonists in therapy. In one embodiment the invention
`relates to certain dosage regimes of GLP-1 agonists which
`provide improved effect in diseases or conditions, such as
`55 prevention and/or treatment of type 2 diabetes and obesity.
`In one embodiment the methods of the present invention
`provides surprisingly showed improved reduction of HbAlc
`and reduction of body weight. In one embodiment the
`GLP-1 agonist is administered in an amount which provides
`an improved a) reduction in HbAlc orb) reduction in body
`weight compared to administration of 1.8 mg liraglutide or
`less, such as 0.8 mg liraglutide or less, per day.
`In one embodiment the invention relates to a method for
`reduction of HbAl c or for prevention or treatment of type 2
`diabetes, hyperglycemia, impaired glucose tolerance, or
`non-insulin dependent diabetes, said method comprising
`administration of a GLP-1 agonist to a subject in need
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00008
`
`

`

`US 10,335,462 B2
`
`3
`thereof in an amount of at least 0.7 mg per week, such an
`amount equivalent to at least 0.7 mg semaglutide per week.
`In one embodiment the method is for reduction of HbAlc.
`In one embodiment the method is for prevention or treat(cid:173)
`ment of type 2 diabetes. In one embodiment the method is
`for prevention or treatment of hyperglycemia. In one
`embodiment the method is for prevention or treatment of
`impaired glucose tolerance. In one embodiment the method
`is for prevention or treatment of non-insulin dependent
`diabetes. In one embodiment the method of the invention 10
`comprises delaying or preventing diabetic disease progres(cid:173)
`sion. In one embodiment a HbAlc level below 7% is
`achieved. In one embodiment the level of HbAlc is deter(cid:173)
`mined according to the method defined by the Diabetes
`Control and Complications Trial (DCCT). In one embodi(cid:173)
`ment the level of HbAlc is determined according to the
`method defined by the International Federation of Clinical
`Chemistry (IFCC).
`In one embodiment the invention relates to a method for
`treating or preventing obesity, for reducing body weight
`and/or food intake, or for inducing satiety, said method
`comprising administration of a GLP-1 agonist to a subject in
`need thereof in an amount of at least 0.7 mg per week, such
`an amount equivalent to at least 0.7 mg semaglutide per
`week. In one embodiment the method is for prevention or 25
`treatment of obesity. In one embodiment the method is for
`reducing body weight and/or food intake. In one embodi(cid:173)
`ment the method is for inducing satiety.
`In one embodiment the GLP-1 agonist has a half-life of at
`least 24 hours, such as at least 48 hours, at least 60 hours, 30
`or at least 72 hours, or such as at least 84 hours, at least 96
`hours, or at least 108 hours, or optionally at least 120 hours,
`at least 132 hours, or at least 144 hours, wherein said
`half-life optionally is determined by Assay (II).
`In one embodiment the GLP-1 agonist is administered 35
`twice weekly or less often, once weekly or less often, or
`once weekly or less often. In one embodiment the GLP-1
`agonist is administered once every secondly week or less
`often, once every third week or less often, or once a month
`or less often.
`In one embodiment the GLP-1 agonist is administered in
`an amount per week of at least 0.8 mg, at least 0.9 mg, or at
`least 1.0 mg. In one embodiment the GLP-1 agonist is
`administered in an amount per week of at least 1.1 mg, at
`least 1.2 mg, or at least 1.3 mg. In one embodiment the 45
`GLP-1 agonist is administered in an amount per week of at
`least 1.4 mg, at least 1.5 mg, or at least 1.6 mg.
`In one embodiment the GLP-1 agonist is administered in
`an amount per week equivalent to at least 0.8 mg, at least 0.9
`mg, or at least 1.0 mg semaglutide. In one embodiment the 50
`GLP-1 agonist is administered in an amount per week
`equivalent to at least 1.1 mg, at least 1.2 mg, or at least 1.3
`mg semaglutide. In one embodiment the GLP-1 agonist is
`administered in an amount per week equivalent to at least
`1.4 mg, at least 1.5 mg, or at least 1.6 mg semaglutide.
`In one embodiment the GLP-1 agonist is selected from the
`group consisting of semaglutide, exenatide, albiglutide, and
`dulaglutide.
`In one embodiment the GLP-1 agonist is administered by
`parenteral administration, such as subcutaneous injection.
`In one embodiment the GLP-1 agonist is a GLP-1 peptide.
`In one embodiment the GLP-1 peptide comprises no more
`than 5, such as no more than 4 or no more than 3, amino acid
`residues which have been substituted, inserted or deleted as
`compared to GLP-1 (7-37). In one embodiment the GLP-1 65
`peptide comprises no more than 4 amino acid residues which
`are not encoded by the genetic code.
`
`4
`In one embodiment the GLP-1 peptide is a DPPIV pro(cid:173)
`tected GLP-1 peptide. In one embodiment the GLP-1 pep(cid:173)
`tide is DPPIV stabilised.
`In one embodiment the GLP-1 agonist has an EC50 at or
`below 3000 pM, such as at or below 500 pM or at or below
`100 pM, optionally determined by Assay (I).
`In one embodiment the invention relates to a GLP-1
`agonist for use in the reduction of HbAlc or for use in the
`prevention or treatment of type 2 diabetes, hyperglycemia,
`impaired glucose tolerance, or non-insulin dependent dia(cid:173)
`betes comprising administering a GLP-1 agonist in an
`amount of at least 0.7 mg per week, such an amount
`equivalent to at least 0.7 mg semaglutide per week. In one
`15 embodiment the GLP-1 agonist and/or administration is as
`defined herein.
`In one embodiment the invention relates to a GLP-1
`agonist for use in the prevention or treatment of obesity, in
`the reduction of body weight and/or food intake, or in the
`20 induction satiety comprising administering a GLP-1 agonist
`in an amount of at least 0.7 mg per week, such an amount
`equivalent to at least 0.7 mg semaglutide per week. In one
`embodiment the GLP-1 agonist and/or administration is as
`defined herein.
`In one embodiment the invention relates to a composition
`comprising a GLP-1 agonist and one or more pharmaceuti(cid:173)
`cally acceptable excipients for use in reduction of HbAlc or
`for prevention or treatment of type 2 diabetes, hyperglyce(cid:173)
`mia, impaired glucose tolerance, or non-insulin dependent
`diabetes, wherein said GLP-1 agonist is administered in an
`amount of at least 0.7 mg per week, such an amount
`equivalent to at least 0.7 mg semaglutide per week. In one
`embodiment the GLP-1 agonist and/or administration is as
`defined herein.
`In one embodiment the invention relates to a composition
`comprising a GLP-1 agonist and one or more pharmaceuti(cid:173)
`cally acceptable excipients for use in the prevention or
`treatment of obesity, in the reduction of body weight and/or
`food intake, or in the induction satiety, wherein said GLP-1
`40 agonist is administered in an amount of at least 0.7 mg per
`week, such an amount equivalent to at least 0.7 mg sema(cid:173)
`glutide per week. In one embodiment the GLP-1 agonist
`and/or administration is as defined herein.
`In one embodiment the GLP-1 agonist is administered
`with another therapeutic agent. Administration with another
`therapeutic agent may be carried out as administration of the
`GLP-1 agonist and the other therapeutic agent within the
`same therapeutic window ( e.g. within a period of two weeks,
`a period of one week, or in a 96, 72, or 48 hour period, etc.).
`The treatment with a GLP-1 agonist according to the present
`invention may be combined with one or more additional
`therapeutic agents, e.g. selected from antidiabetic agents,
`antiobesity agents, appetite regulating agents, antihyperten(cid:173)
`sive agents, agents for the treatment and/or prevention of
`55 complications resulting from or associated with diabetes and
`agents for the treatment and/or prevention of complications
`and disorders resulting from or associated with obesity;
`examples of these therapeutic agents are: sulphonylureas,
`thiazolidinediones, biguanides, meglitinides, glucosidase
`60 inhibitors, glucagon antagonists, and DPP-IV (dipeptidyl
`peptidase-IV) inhibitors.
`In one embodiment, as used herein, an "amount equiva-
`lent to" when used in relation to GLP-1 agonists refers to
`amounts of a first GLP-1 agonist and a second GLP-1
`agonist having GLP-1 receptor potency (i.e. EC50) within
`±30%, such as within ±20% or within ±10%, of each other
`optionally determined by Assay (I) described herein and
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00009
`
`

`

`US 10,335,462 B2
`
`5
`having a half-life within ±30%, such as within ±20% or
`within ±10%, of each other optionally determined by Assay
`(II) described herein.
`In one embodiment an "effective amount" of a GLP-1
`agonist as used herein means an amount sufficient to cure,
`alleviate, or partially arrest the clinical manifestations of a
`given disease or state and its complications. An amount
`adequate to accomplish this is defined as "effective amount".
`Effective amounts for each purpose will depend on the
`severity of the disease or injury as well as the weight and
`general state of the subject. It will be understood that
`determining an appropriate dosage may be achieved using
`routine experimentation, by constructing a matrix of values
`and testing different points in the matrix, which is all within
`the ordinary skills of a trained physician or veterinary.
`In one embodiment the term "treatment" or "treating" as
`used herein means the management and care of a patient for
`the purpose of combating a condition, such as a disease or
`a disorder. In one embodiment the term "treatment" or
`"treating" is intended to include the full spectrum of treat(cid:173)
`ments for a given condition from which the patient is
`suffering, such as administration of the active compound to
`alleviate the symptoms or complications; to delay the pro(cid:173)
`gression of the disease, disorder, or condition; to alleviate or
`relieve the symptoms and complications; and/or, to cure or
`eliminate the disease, disorder, or condition as well as to
`prevent the condition. In one embodiment prevention is to be
`understood as the management and care of a patient for the
`purpose of combating the disease, condition, or disorder and
`includes the administration of the active compounds to
`prevent the onset of the symptoms or complications.
`In one embodiment the term "hydrophilic spacer" as used
`herein means a spacer that separates a peptide and an
`albumin binding residue with a chemical moiety which
`comprises at least 5 non-hydrogen atoms where 30-50% of
`these are either N or 0.
`In one embodiment the term "analogue" as used herein
`referring to a polypeptide means a modified peptide wherein
`one or more amino acid residues of the peptide have been
`substituted by other amino acid residues and/or wherein one
`or more amino acid residues have been deleted from the
`peptide and or wherein one or more amino acid residues
`have been added to the peptide. Such addition or deletion of
`amino acid residues can take place at the N-terminal of the
`peptide and/or at the C-terminal of the peptide. A simple
`system
`is used to describe analogues: For example
`Arg34GLP-1 (7-37) Lys designates a GLP-1 analogue
`wherein the naturally occurring lysine at position 34 has
`been substituted with arginine and a lysine residue has been
`added to the C-terminal (position 38).
`In one embodiment the term "GLP-1 peptide" as used
`herein means GLP-1 (7-37), a GLP-1 analogue, a GLP-1
`derivative or a derivative of a GLP-1 analogue.
`In one embodiment the term "exendin-4 peptide" as used
`herein means exendin-4 (1-39), an exendin-4 analogue, an
`exendin-4 derivative or a derivative of an exendin-4 ana(cid:173)
`logue.
`In one embodiment the term "DPP-IV protected" as used
`herein referring to a polypeptide means a polypeptide which
`has been chemically modified in order to render said com(cid:173)
`pound resistant to the plasma peptidase dipeptidyl amino(cid:173)
`peptidase-4 (DPP-IV). The DPP-IV enzyme in plasma is
`known to be involved in the degradation of several peptide
`hormones, e.g. GLP-1, Exendin-4 etc. Thus a considerable
`effort is being made to develop GLP-1 agonists less suscep(cid:173)
`tible to DPP-IV mediated hydrolysis in order to reduce the
`rate of degradation by DPP-IV.
`
`10
`
`6
`The present invention also relates to a GLP-1 agonist of
`the invention, for use as a medicament. In particular embodi(cid:173)
`ments, the GLP-1 agonist of the invention may be used for
`the following medical treatments:
`(i) prevention and/or treatment of all forms of diabetes,
`such as hyperglycemia, type 2 diabetes, impaired glucose
`tolerance, type 1 diabetes, non-insulin dependent diabetes,
`MODY (maturity onset diabetes of the young), gestational
`diabetes, and/or for reduction of HbAlc;
`(ii) delaying or preventing diabetic disease progression,
`such as progression in type 2 diabetes, delaying the pro(cid:173)
`gression of impaired glucose tolerance (IGT) to insulin
`requiring type 2 diabetes, and/or delaying the progression of
`15 non-insulin requiring type 2 diabetes to insulin requiring
`type 2 diabetes;
`(iii) prevention and/or treatment of eating disorders, such
`as obesity, e.g. by decreasing food intake, reducing body
`weight, suppressing appetite, inducing satiety; treating or
`20 preventing binge eating disorder, bulimia nervosa, and/or
`obesity induced by administration of an antipsychotic or a
`steroid; reduction of gastric motility; and/or delaying gastric
`emptying.
`In another particular embodiment, the indication is (i). In
`25 a further particular embodiment the indication is (ii). In a
`still further particular embodiment the indication is (iii). In
`one embodiment the indication is type 2 diabetes and/or
`obesity.
`In one embodiment the method comprises prevention,
`30 treatment, reduction and/or induction in one or more dis(cid:173)
`eases or conditions defined herein. In one embodiment the
`indication is (i) and (iii). In one embodiment the indication
`is (ii) and (iii). In one embodiment the method comprises
`prevention, treatment, reduction and/or induction in one or
`35 more diseases or conditions selected from a) and b ), a) and
`c ), b) and c ), or a), b) and c) as defined in claim 1.
`In one embodiment the invention relates to administration
`of an effective amount of a GLP-1 agonist.
`In one embodiment as used herein, specific values given
`40 in relation to numbers or intervals may be understood as the
`specific value or as about the specific value.
`Functional Properties
`In a first functional aspect, the GLP-1 agonists of the
`invention have a good potency. Also, or alternatively, in a
`45 second functional aspect, the GLP-1 agonists of the inven(cid:173)
`tion have a protracted pharmacokinetic profile. Also, or
`alternatively, in a third functional aspect, the GLP-1 agonists
`of the invention are stable against degradation by gastro
`intestinal enzymes.
`50 Biological Activity (Potency)
`According to the first functional aspect, the GLP-1 ago(cid:173)
`nists of the invention are biologically active, or potent. In a
`particular embodiment, "potency" and/or "activity" refers to
`in vitro potency, i.e. performance in a functional GLP-1
`55 receptor assay, more in particular to the capability of stimu(cid:173)
`lating cAMP formation in a cell line expressing the cloned
`human GLP-1 receptor.
`The stimulation of the formation of cAMP in a medium
`containing the human GLP-1 receptor may preferably be
`60 determined using a stable transfected cell-line such as
`BHK467-12A (tk-ts13), and/or using for the determination
`of cAMP a functional receptor assay, e.g. based on compe(cid:173)
`tition between endogenously formed cAMP and exog(cid:173)
`enously added biotin-labelled cAMP, in which assay cAMP
`65 is more preferably captured using a specific antibody, and/or
`wherein an even more preferred assay is the AlphaScreen
`cAMP Assay, such as the one described in Assay (I).
`
`Novo Nordisk Exhibit 2003
`Mylan Pharms. Inc.v. Novo Nordisk A/S
`IPR2023-00722
`Page 00010
`
`

`

`H-His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser(cid:173)
`Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu
`Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-
`Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2
`(SEQ ID NO: 2).
`
`In one embodiment the GLP-1 agonist comprises an
`albumin binding residue attached via a hydrophilic spacer to
`the C-terminal amino acid residue of said GLP-1 peptide.
`In one embodiment the GLP-1 agonist comprises a second
`albumin binding residue is attached to an amino acid residue
`which is not the C-terminal amino acid residue.
`In one embodiment the GLP-1 peptide is selected from
`the group consisting of semaglutide, albiglutide and dula(cid:173)
`glitide.
`In one embodiment the GLP-1 peptide has the following
`structure:
`
`30
`
`(His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser(cid:173)
`Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-
`Ala-Trp-Leu-Val-Lys-Gly-Arg)2
`(SEQ ID NO: 4)
`
`US 10,335,462 B2
`
`8
`In one embodiment the amino acid residue in position 7
`of said GLP-1 peptide is selected from the group consisting
`ofD-histidine, desamino-histidine, 2-amino-histidine, ~-hy(cid:173)
`droxy-histidine, homohistidine, N"'-acetyl-histidine, a-fluo(cid:173)
`romethy I-histidine, a-methy I-histidine, 3-pyridylalanine,
`2-pyridylalanine and 4-pyridylalanine.
`In one embodiment the GLP-1 peptide is attached to a
`hydrophilic spacer via the amino acid residue in position 23,
`26, 34, 36 or 38 relative to the amino acid sequence of
`GLP-1 (7-37).
`In one embodiment the GLP-1 peptide is exendin-4, an
`exendin-4-analogue, or a derivative of exendin-4.
`In one embodiment the GLP-1 agonist peptide comprises
`the amino acid sequence of the following formula:
`
`7
`In one embodiment the term half maximal effective
`concentration (EC50) generally refers to the concentration
`which induces a response halfway between the baseline and
`maximum, by reference to the dose response curve. EC50 is
`used as a measure of the potency of a compound and
`represents the concentration where 50% of its maximal
`effect is observed.
`The in vitro potency of the GLP-1 agonists of the inven(cid:173)
`tion may be determined as described above, and the EC50 of
`the GLP-1 agonist in question determined. The lower the 10
`EC50 , the better the potency.
`In a particular embodiment, the medium has the following
`composition (final in-assay concentrations): 50 mM TRIS(cid:173)
`HCI; 5 mM HEPES; 10 mM MgCl 2 , 6H2 O; 150 mM NaCl; 15
`0.01%Tween; 0.1%BSA; 0.5mMIBMX; 1 mMATP; 1 µM
`GTP; pH 7.4.
`In a further particular embodiment, the GLP-1 agonist of
`the invention has an in vitro potency corresponding to an
`EC50 at or below 3000 pM, such as below 2000 pM, below 20
`1000 pM, or below 500 pM, or such as below 200 pM or
`below 100 pM.
`In another particular embodiment the GLP-1 agonist of
`the invention are potent in vivo, which may be determined
`as is known in the art in any suitable animal model, as well 25
`as in clinical trials.
`The diabetic db/db mouse is one example of a suitable
`animal model, and the blood glucose lowering effect may be
`determined in such mice in vivo, e.g. as described in Assay
`(III), or as described in Example 43 of WO09/030738.
`Also, or alternatively, the effect on food intake in vivo
`His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-
`may be determined in pharma