`
`US008536122B2
`
`c12) United States Patent
`Lau et al.
`
`(IO) Patent No.:
`(45) Date of Patent:
`
`US 8,536,122 B2
`*Sep.17,2013
`
`(54) ACYLATED GLP-1 COMPOUNDS
`
`(75)
`
`Inventors: Jesper Lau, Farum (DK); Florencio
`Zaragoza Doerwald, Smorum (DK);
`Paw Bloch, Taastrup (DK); Thomas
`Kruse Hansen, Herlev (DK)
`
`(73) Assignee: Novo NordiskA/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.: 13/412,283
`
`(22) Filed:
`
`Mar. 5, 2012
`
`(65)
`
`Prior Publication Data
`
`US 2012/0295847 Al
`
`Nov. 22, 2012
`
`Related U.S. Application Data
`
`(63) Continuation of application No. 11/908,834, filed as
`application No. PCT/EP2006/060855 on Mar. 20,
`2006, now Pat. No. 8,129,343.
`
`(60) Provisional application No. 60/664,497, filed on Mar.
`23, 2005.
`
`(30)
`
`Foreign Application Priority Data
`
`Mar. 18, 2005
`
`(EP) ..................................... 05102171
`
`(51)
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`Int. Cl.
`A61K 38/26
`A61K 38/28
`A61P3/10
`A61P 7112
`C07K 141605
`C07K5/00
`C07K 7100
`C07K 16100
`C07K 17100
`(52) U.S. Cl.
`USPC ............................ 514/7.2; 514/11.7; 530/308
`( 58) Field of Classification Search
`None
`See application file for complete search history.
`
`(56)
`
`References Cited
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`9/2006
`2006/097537 A2
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`
`(Continued)
`Primary Examiner - Marcela M Cordero Garcia
`(74) Attorney, Agent, or Firm - Richard W. Bork
`ABSTRACT
`(57)
`Protracted GLP-1 compounds and therapeutic uses thereof.
`16 Claims, No Drawings
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00001
`
`
`
`US 8,536,122 B2
`Page 2
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`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00002
`
`
`
`US 8,536,122 B2
`
`1
`ACYLATED GLP-1 COMPOUNDS
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`5
`
`This application is a Continuation of copending U.S. appli(cid:173)
`cation Ser. No. 11/908,834, filed Sep. 17, 2007, which is a 35
`U.S.C. §371 national stage application oflnternational Patent
`Application PCT/EP2006/060855 (published as WO 2006/
`097537), filed Mar. 20, 2006, which claimed priority of Euro- 10
`peanPatentApplication 05102171.5, filed Mar. 18, 2005; this
`application further claims priority under 35 U.S.C. §119 of
`U.S. Provisional Application 60/664,497, filed Mar. 23, 2005.
`
`2
`invention and the use of compounds according to the present
`invention for preparing medicaments for treating disease.
`The invention provides a method for increasing the time of
`action in a patient of a GLP-1 analog, characterised in acy-
`lating said GLP-1 analog with a moiety B-U' as disclosed in
`any of the preceding claims, on the lysine residue in position
`26 of said GLP-1 analog.
`
`DESCRIPTION OF THE INVENTION
`
`FIELD OF THE INVENTION
`
`This invention relates to the field of therapeutic peptides,
`i.e. to new protracted GLP-1 compounds.
`
`BACKGROUND OF THE INVENTION
`
`A range of different approaches have been used for modi(cid:173)
`fying the structure of glucagon-like peptide 1 (GLP-1) com(cid:173)
`pounds in order to provide a longer duration of action in vivo.
`WO 96/29342 discloses peptide hormone derivatives wherein
`the parent peptide hormone has been modified by introducing
`a lipophilic substituent in the C-terminal amino acid residue
`or in the N-terminal amino acid residue.
`WO 98/08871 discloses GLP-1 derivatives wherein at least
`one amino acid residue of the parent peptide has a lipophilic
`substituent attached.
`WO 99/43708 discloses GLP-1(7-35) and GLP-1(7-36)
`derivatives which have a lipophilic substituent attached to the
`C-terminal amino acid residue.
`WO 00/34331 discloses acylated GLP-1 analogs.
`WO 00/69911 discloses activated insulinotropic peptides
`to be injected into patients where they are supposed to react
`with blood components to form conjugates and thereby alleg(cid:173)
`edly providing longer duration of action in vivo.
`WO 02/46227 discloses GLP-1 and exendin-4 analogs
`fused to human serum albumin in order to extend in vivo
`half-life.
`Many diabetes patients particularly in the type 2 diabetes
`segment are subject to so-called "needle-phobia", i.e. a sub(cid:173)
`stantial fear of injecting themselves. In the type 2 diabetes
`segment most patients are treated with oral hypoglycaemic
`agents, and since GLP-1 compounds are expected to be the
`first injectable product these patients will be administered, the
`fear of injections may become a serious obstacle for the
`widespread use of the clinically very promising GLP-1 com(cid:173)
`pounds. Thus, there is a need to develop new GLP-1 com(cid:173)
`pounds which can be administered less than once daily, e.g.
`once every second or third day preferably once weekly, while
`retaining an acceptable clinical profile.
`
`SUMMARY OF THE INVENTION
`
`The invention provides a GLP-1 analog having a modifi(cid:173)
`cation of at least one non-proteogenic amino acid residue in
`positions 7 and/or 8 relative to the sequence GLP-1(7-37)
`(SEQ ID No 1 ), which is acylated with a moiety to the lysine
`residue in position 26, and where said moiety comprises at
`least two acidic groups, wherein one acidic group is attached
`terminally.
`The present invention also provides pharmaceutical com(cid:173)
`positions comprising a compound according to the present
`
`In the present specification, the following terms have the
`indicated meaning:
`The term "polypeptide" and "peptide" as used herein
`means a compound composed of at least five constituent
`15 amino acids connected by peptide bonds. The constituent
`amino acids may be from the group of the amino acids
`encoded by the genetic code and they may be natural amino
`acids which are not encoded by the genetic code, as well as
`synthetic amino acids. Natural amino acids which are not
`20 encoded by the genetic code are e.g., y-carboxyglutamate,
`omithine, phosphoserine, D-alanine and D-glutamine. Syn(cid:173)
`thetic amino acids comprise amino acids manufactured by
`chemical synthesis, i.e. D-isomers of the amino acids
`encoded by the genetic code such as D-alanine and D-leucine,
`25 Aib (a-aminoisobutyric acid), Abu (a-aminobutyric acid),
`Tie (tert-butylglycine), ~-alanine, 3-aminomethyl benzoic
`acid, anthranilic acid.
`The 22 proteogenic amino acids are:
`Alanine, Arginine, Asparagine, Aspartic acid, Cysteine, Cys-
`30 tine, Glutamine, Glutamic acid, Glycine, Histidine, Hydrox(cid:173)
`yproline, Isoleucine, Leucine, Lysine, Methionine, Phenyla(cid:173)
`lanine, Praline, Serine, Threonine, Tryptophan, Tyrosine,
`Valine.
`Thus a non-proteogenic amino acid is a moiety which can
`35 be incorporated into a peptide via peptide bonds but is not a
`proteogenic amino acid. Examples are y-carboxyglutamate,
`omithine, phosphoserine, the D-amino acids such as D-ala(cid:173)
`nine and D-glutamine, Synthetic non-proteogenic amino
`acids comprise amino acids manufactured by chemical syn-
`40 thesis, i.e. D-isomers of the amino acids encoded by the
`genetic code such as D-alanine and D-leucine, Aib (a-ami(cid:173)
`noisobutyric acid), Abu (a-aminobutyric acid), Tie (tert-bu(cid:173)
`tylglycine), 3-aminomethyl benzoic acid, anthranilic acid,
`des-amino-Histidine, the beta analogs of amino acids such as
`45 ~-alanine etc. D-histidine, desamino-histidine, 2-amino-his(cid:173)
`tidine, ~-hydroxy-histidine, homohistidine, N"'-acetyl-histi(cid:173)
`dine, a-fluoromethyl-histidine, a-methyl-histidine, 3-py(cid:173)
`ridylalanine,
`2-pyridylalanine
`or
`4-pyridylalanine,
`(1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl)
`50 carboxylic acid, (1-aminocyclopentyl) carboxylic acid,
`(1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl)
`carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
`The term "analogue" as used herein referring to a polypep(cid:173)
`tide means a modified peptide wherein one or more amino
`55 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 deleted from the
`peptide and or wherein one or more amino acid residues have
`60 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
`often used to describe analogues: For example [Arg34]GLP-
`1(7-37)Lys designates a GLP-1(7-37) analogue wherein the
`65 naturally occurring lysine at position 34 has been substituted
`with arginine and wherein a lysine has been added to the
`terminal amino acid residue, i.e. to the Gly37
`. All amino acids
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00003
`
`
`
`US 8,536,122 B2
`
`3
`for which the optical isomer is not stated is to be understood
`to mean the L-isomer. In embodiments of the invention a
`maximum of 1 7 amino acids have been modified. In embodi(cid:173)
`ments of the invention a maximum of 15 amino acids have
`been modified. In embodiments of the invention a maximum 5
`of 10 amino acids have been modified. In embodiments of the
`invention a maximum of 8 amino acids have been modified. In
`embodiments of the invention a maximum of 7 amino acids
`have been modified. In embodiments of the invention a maxi(cid:173)
`mum of 6 amino acids have been modified. In embodiments 10
`of the invention a maximum of 5 amino acids have been
`modified. In embodiments of the invention a maximum of 4
`amino acids have been modified. In embodiments of the
`invention a maximum of3 amino acids have been modified. In
`embodiments of the invention a maximum of 2 amino acids 15
`have been modified. In embodiments of the invention 1 amino
`acid has been modified.
`The term "derivative" as used herein in relation to a peptide
`means a chemically modified peptide or an analogue thereof,
`wherein at least one substituent is not present in the unmodi- 20
`fled peptide or an analogue thereof, i.e. a peptide which has
`been covalently modified. Typical modifications are amides,
`carbohydrates, alkyl groups, acyl groups, esters and the like.
`An example of a derivative of GLP-1(7-37) is NE26
`-(( 4S)-4-
`(hexadecanoylamino )-carboxy-butanoyl)[ Arg34
`Lys 26
`] 25
`,
`GLP-1-(7-37).
`The term "GLP-1 peptide" as used herein means GLP-1(7-
`37) (SEQ ID No 1), a GLP-1(7-37) analogue, a GLP-1(7-37)
`derivative or a derivative of a GLP-1(7-37) analogue. In one
`embodiment the GLP-1 peptide is an insulinotropic agent.
`The term "insulinotropic agent" as used herein means a
`compound which is an agonist of the human GLP-1 receptor,
`i.e. a compound which stimulates the formation of cAMP in
`a suitable medium containing the human GLP-1 receptor ( one
`such medium disclosed below). The potency of an insulino- 35
`tropic agent is determined by calculating the EC50 value from
`the dose-response curve as described below.
`Baby hamster kidney (BHK) cells expressing the cloned
`human GLP-1 receptor (BHK-467-12A) were grown in
`DMEM media with the addition of 100 IU/mL penicillin, 100 40
`µg/mL streptomycin, 5% fetal calf serum and 0.5 mg/mL
`Geneticin G-418 (Life Technologies). The cells were washed
`twice in phosphate buffered saline and harvested with
`Versene. Plasma membranes were prepared from the cells by
`homogenisation with an Ultraturrax in buffer 1 (20 mM 45
`HEPES-Na, 10 mM EDTA, pH 7.4). The homogenate was
`centrifuged at 48,000xg for 15 min at 4 ° C. The pellet was
`suspended by homogenization in buffer 2 (20 mM HEPES-
`Na, 0.1 mM EDTA, pH 7.4 ), then centrifuged at 48,000xg for
`15 min at 4 ° C. The washing procedure was repeated one 50
`more time. The final pellet was suspended in buffer 2 and used
`immediately for assays or stored at -80° C.
`The functional receptor assay was carried out by measur-
`ing cyclic AMP ( cAMP) as a response to stimulation by the
`insulinotropic agent. cAMP formed was quantified by the 55
`AlphaScreen™ cAMP Kit (Perkin Elmer Life Sciences).
`Incubations were carried out in half-area 96-well microtiter
`plates ina total volume of50 µL buffer3 (50mMTris-HCl, 5
`mM HEPES, 10 mM MgC12 , pH 7.4) and with the following
`additions: 1 mM ATP, 1 µM GTP, 0.5 mM 3-isobutyl-1- 60
`methylxanthine (IBMX), 0.01 % Tween-20, 0.1 % BSA, 6 µg
`membrane preparation, 15 µg/mL acceptor beads, 20 µg/mL
`donor beads preincubated with 6 nM biotinyl-cAMP. Com(cid:173)
`pounds to be tested for agonist activity were dissolved and
`diluted in buffer 3. GTP was freshly prepared for each experi- 65
`ment. The plate was incubated in the dark with slow agitation
`for three hours at room temperature followed by counting in
`
`4
`the Fusion™ instrument (Perkin Elmer Life Sciences). Con(cid:173)
`centration-response curves were plotted for the individual
`compounds and EC50 values estimated using a four-param-
`eter logistic model with Prism v. 4.0 (GraphPad, Carlsbad,
`Calif.).
`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 compound resistant to the
`plasma peptidase dipeptidyl aminopeptidase-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, GLP-2,
`Exendin-4 etc. Thus, a considerable effort is being made to
`develop analogues and derivatives of the polypeptides sus(cid:173)
`ceptible to DPP-IV mediated hydrolysis in order to reduce the
`rate of degradation by DPP-IV. In one embodiment a DPP-IV
`protected peptide is more resistant to DPP-IV than GLP-1(7-
`37) or Exendin-4(1-39).
`Resistance of a peptide to degradation by dipeptidyl ami(cid:173)
`nopeptidase IV is determined by the following degradation
`assay:
`Aliquots of the peptide (5 nmol) are incubated at 37° C.
`with 1 µL of purified dipeptidyl aminopeptidase IV corre(cid:173)
`sponding to an enzymatic activity of 5 m U for 10-180 minutes
`in 100 µL of0.1 M triethylamine-HCl buffer, pH 7.4. Enzy(cid:173)
`matic reactions are terminated by the addition of 5 µL of 10%
`trifluoroacetic acid, and the peptide degradation products are
`separated and quantified using HPLC analysis. One method
`for performing this analysis is: The mixtures are applied onto
`a Vydac C18 widepore (30 nm pores, 5 µm particles) 250x4.6
`30 mm colunm and eluted at a flow rate of 1 ml/min with linear
`stepwise gradients of acetonitrile in 0.1 % trifluoroacetic acid
`(0% acetonitrile for 3 min, 0-24% acetonitrile for 17 min,
`24-48% acetonitrile for 1 min) according to Siegel et al.,
`Regul. Pept. 1999; 79:93-102 and Mentlein et al. Eur. J.
`Biochem. 1993; 214:829-35. Peptides and their degradation
`products may be monitored by their absorbance at 220 nm
`(peptide bonds) or 280 nm (aromatic amino acids), and are
`quantified by integration of their peak areas related to those of
`standards. The rate of hydrolysis of a peptide by dipeptidyl
`aminopeptidase IV is estimated at incubation times which
`result in less than 10% of the peptide being hydrolysed.
`The term "C 1_6 -alkyl" as used herein means a saturated,
`branched, straight or cyclic hydrocarbon group having from 1
`to 6 carbon atoms. Representative examples include, but are
`not limited to, methyl, ethyl, n-propyl, isopropyl, butyl,
`isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl,
`tert-pentyl, n-hexyl, isohexyl, cyclohexane and the like. The
`term "pharmaceutically acceptable" as used herein means
`suited for normal pharmaceutical applications, i.e. giving rise
`to no adverse events in patients etc.
`The term "excipient" as used herein means the chemical
`compounds which are normally added to pharmaceutical
`compositions, e.g. buffers, tonicity agents, preservatives and
`the like.
`The term "effective amount" as used herein means a dosage
`which is sufficient to be effective for the treatment of the
`patient compared with no treatment.
`The term "pharmaceutical composition" as used herein
`means a product comprising an active compound or a salt
`thereof together with pharmaceutical excipients such as
`buffer, preservative, and optionally a tonicity modifier and/or
`a stabilizer. Thus a pharmaceutical composition is also known
`in the art as a pharmaceutical formulation.
`The term "treatment of a disease" as used herein means the
`management and care of a patient having developed the dis(cid:173)
`ease, condition or disorder. The purpose of treatment is to
`combat the disease, condition or disorder. Treatment includes
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00004
`
`
`
`US 8,536,122 B2
`
`In the formulas below
`
`6
`
`5
`the administration of the active compounds to eliminate or
`control the disease, condition or disorder as well as to allevi(cid:173)
`ate the symptoms or complications associated with the dis(cid:173)
`ease, condition or disorder.
`In another aspect the present invention relates to an acy-
`lated GLP-1 analogue that can bind to albumin and the GLP-1
`receptor simultaneously.
`In another aspect the present invention relates to an acy(cid:173)
`lated GLP-1 analogue that bind to the GLP-1 receptor with an
`affinity below 100 nM, preferable below 30 nM in the pres- 10
`ence of 2% albumin.
`In another aspect the present invention relates to an acy(cid:173)
`lated GLP-1 analogue which affinity to the GLP-1 receptor is
`only partly decreased when comparing the affinity in the 15
`presence of very low concentration (e.g. 0.005% to 0.2%) of
`human albumin to the affinity in the presence of 2% human
`albumin. The shift in binding affinity under these conditions
`is less than 50 fold, preferable below 30 fold and more pref(cid:173)
`erable below 10 fold.
`The term "albumin binding moiety" as used herein means
`a residue which binds non-covalently to human serum albu(cid:173)
`min. The albumin binding residue attached to the therapeutic
`polypeptide typically has an affinity below 10 µM to human
`serum albumin and preferably below 1 µM. A range of albu(cid:173)
`min binding residues are known among linear and branched
`lipohophillic moieties containing 4-40 carbon atoms having a
`distal acidic group.
`The term "hydrophilic linker" as used herein means a
`spacer that separates a peptide and an albumin binding resi(cid:173)
`due with a chemical moiety which comprises at least 5 non(cid:173)
`hydrogen atoms where 30-50% of these are either Nor 0.
`The term "acidic groups" as used herein means organic
`chemical groups which are fully or partly negatively charged
`at physiological pH. The pKa value of such groups is below 7,
`preferable below 5. This includes but is not limited to car(cid:173)
`boxylic acids, sulphonic acids, phosphoric acids or heterocy(cid:173)
`clic ring systems which are fully or partly negatively charged
`at physiological pH.
`In the below structural formula II the moiety U is a di- 40
`radical may be attached to the terminal groups B and the
`aminogroup of the lysine amino acid in the peptide in two
`different ways. In embodiments of the invention the U in
`formula II is attached with the group B attached at the end of
`the alkyl chain and the peptide at the other end.
`In the formulas below the terminal bonds from the attached
`groups are to be regarded as attachment bonds and not ending
`in methylene groups unless stated.
`
`5
`
`20
`
`means the H2N-His-Aib-N-terminal of the GLP-1 analogue.
`In an embodiment the invention provides a GLP-1 analog
`acylated with a lipophillic albumin binding moiety contain(cid:173)
`ing at least two free acidic chemical groups attached via a non
`natural amino acid linker to the lysine residue in position 26.
`In an embodiment, the term free acidic chemical groups is
`to be understood as having the same meaning as "acidic
`groups" as used herein.
`In an embodiment the invention provides an acylated
`GLP-1 analog where said GLP-1 analog is stabilised against
`DPP-IV by modification of at least one amino acid residue in
`positions 7 and 8 relative to the sequence GLP-1(7-37) (SEQ
`ID No 1 ), and where said acylation is a diacid attached to the
`lysine residue in position 26 optionally via a non natural
`amino acid hydrophilic linker.
`In an embodiment of the invention a GLP-1 analog having
`25 a modification of at least one non-proteogenic amino acid
`residue in positions 7 and/or 8 relative to the sequence GLP-
`1(7-37) (SEQ ID No 1), which is acylated withamoietytothe
`lysine residue in position 26, and where said moiety com(cid:173)
`prises at least two acidic groups, wherein one acidic group is
`30 attached terminally.
`An embodiment provides a GLP-1 analog according to the
`above embodiment, wherein the moiety attached in position
`26 comprises a hydrophilic linker.
`An embodiment provides a GLP-1 analog according to the
`35 above embodiments, wherein the hydrophilic linker com(cid:173)
`prises at least 5 non-hydrogen atoms where 30-50% of these
`are either Nor 0.
`An embodiment provides a GLP-1 analog according to any
`of the above embodiments, wherein the moiety attached in
`position 26 comprises an albumin binding moiety separated
`from the peptide by the hydrophilic linker.
`An embodiment provides a GLP-1 analog according to the
`above embodiment, wherein the albumin binding moiety is a
`linear or branched lipophilic moiety containing 4-40 carbon
`45 atoms having a distal acidic group.
`An embodiment provides a GLP-1 analog according to any
`of the above embodiments, wherein the acylated moiety is
`B-U', where U' is selected from
`
`0
`HO..._J
`
`YS:::,.._
`
`"-N
`H
`
`O
`
`~~ o~0
`
`0~ o~
`0N~
`'H
`••
`
`II
`0
`
`0
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00005
`
`
`
`US 8,536,122 B2
`
`7
`
`8
`
`,:10H
`H~
`0
`
`/ ij ~o~o~N~o~o~ .
`
`'H
`
`••
`
`II
`0
`
`i j~ ~o~
`,oYoH
`~~ 0
`~( tt l H
`
`0
`
`0
`
`O
`
`p
`
`n
`
`0
`
`~\
`'-.._~/S ....._,,,' 'mlr N~ 0~ 0~ ; : -~ / ' - . . . . . . / 0~ 0~ ,
`
`0
`
`0
`
`mis 0, 1, 2, 3, 4, 5, or 6,
`n is 1, 2 or 3
`s is 0, 1, 2, or 3,
`tis 0, 1, 2, 3, or4
`pis 1,2,3,4,5,6, 7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
`19, 20, 21, 22, or 23;
`and where B is an acidic group selected from
`
`H
`
`HO /N~ and
`
`0
`
`0
`
`55
`
`60
`
`65
`
`-continued
`
`HOD~~
`0
`0
`
`O
`
`HO
`
`0
`
`where 1 is 12, 13, 14, 15, 16, 17, 18, 19 or 20;
`An embodiment provides a GLP-1 analog according to any
`of the above embodiments, which is a compound of formula
`I (SEQ ID No. 2):
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00006
`
`
`
`US 8,536,122 B2
`
`9
`
`10
`
`Formula I
`
`Xaar Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa 16-Ser-Xaa 18-Xaa 19-Xaa20-Glu-Xaa2r
`0
`
`Xaa2r Ala-Xaa25- - ~ ~ -X a a2r Phe-Ile-Xaa30-Trp-Leu-Xaa3rXaa34-Xaa35-Xaa36-Xaa3r
`
`\ B-U'-NH
`
`wherein
`Xaa7 is L-histidine, imidazopropionyl, a-hydroxy-histidine,
`D-histidine, desamino-histidine, 2-amino-histidine, ~-hy(cid:173)
`droxy-histidine,
`homohistidine,
`N"' -acetyl-histidine,
`N"'-formyl-histidine, a-fluoromethyl-histidine, a-methyl(cid:173)
`histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylala(cid:173)
`nine
`Xaa8 is Ala, Gly, Val, Leu, Ile, Thr, Ser, Lys,Aib, (1-aminocy(cid:173)
`clopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic
`acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclo(cid:173)
`hexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic
`acid, or (1-aminocyclooctyl) carboxylic acid;
`Xaa16 is Val or Leu;
`Xaa18 is Ser, Lys or Arg;
`
`15
`
`Xaa19 is Tyr or Gin;
`Xaa20 is Leu or Met;
`Xaa22 is Gly, Glu or Aib;
`Xaa23 is Gin, Glu, Lys or Arg;
`20 Xaa25 is Ala or Val;
`Xaa27 is Glu or Leu;
`Xaa30 is Ala, Glu or Arg;
`Xaa33 is Val or Lys;
`Xaa34 is Lys, Glu, Asn or Arg;
`25 Xaa35 is Gly or Aib;
`Xaa36 is Arg, Gly or Lys, or is absent;
`Xaa37 is Gly, Ala, Glu, Pro, Lys, or is absent;
`and B and U' together is the acylated moiety, where U' is
`selected from
`
`0
`H0,,._11
`
`' - ~Y i j ~ o ~ o ~ ~ ~ o ~ o¾
`
`0
`
`0
`
`0
`
`' -N ~ i j ~ o ~ o 0 ~ ~ 0 ~ 0¾
`
`HOY
`
`O
`
`0
`
`o
`
`, : y i j ~ o ~ o ~ ~ ~ o ~ o¾ ·
`
`0
`
`0
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00007
`
`
`
`US 8,536,122 B2
`
`11
`
`12
`
`30
`
`mis 0, 1, 2, 3, 4, 5, or 6,
`n is 1, 2 or 3
`s is 0, 1, 2, or 3,
`tis 0, 1, 2, 3, or4
`pis 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 35
`19, 20, 21, 22, or 23;
`and where B is an acidic group selected from
`
`-continued
`
`HOD~ y y
`0
`0
`
`O
`
`HO
`
`0
`
`HO~ , ~~ 40
`II
`, [I
`II
`II
`
`0
`
`0
`
`O
`
`O
`
`HO/
`
`I
`
`and
`
`where I is 12, 13, 14, 15, 16, 17, 18, 19 or 20;
`In an embodiment the invention provides a compound
`which is a compound of formula II (SEQ ID No, 3):
`
`Xaar Xaas-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa 16-Ser-Xaa 1s-Xaa 19Xaa20-Glu-Xaa22-
`
`Formula II
`
`0
`
`XM0 -A<,-XM,,-ij~-XM,, ,.,_.,, XM,o T,c T =-XM;, XM,e-XM,,-XM;oXM,c XM;s
`
`\ B-UyNH
`
`0
`
`Novo Nordisk Exhibit 2488
`Mylan Pharms. Inc. v. Novo Nordisk A/S
`IPR2023-00724
`Page 00008
`
`
`
`US 8,536,122 B2
`
`13
`The formula II is identica