`a2) Patent Application Publication co) Pub. No.: US 2007/0010424 Al
`(43) Pub. Date: Jan. 11, 2007
`
`Pedersen et al.
`
`US 20070010424A1
`
`(54)
`
`(75)
`
`PROPYLENE GLYCOL-CONTAINING
`PEPTIDE FORMULATIONS WHICH ARE
`OPTIMAL FOR PRODUCTION AND FOR
`USE IN INJECTION DEVICES
`
`Inventors: Tina Bjeldskov Pedersen, Smorum
`(DK); Claude Bonde, Lyngby (DK);
`Dorthe Kot Engelund, Holte (DK)
`
`Correspondence Address:
`NOVO NORDISK, INC.
`PATENT DEPARTMENT
`100 COLLEGE ROAD WEST
`
`PRINCETON, NJ 08540 (US)
`
`(30)
`
`Foreign Application Priority Data
`
`Nov. 20, 2003
`
`(DK)... eee PA 2003 01719
`
`Publication Classification
`
`(51)
`
`Int. CL
`AGIK 38/28
`
`(2006.01)
`(2006.01)
`AGIK 31/045
`(2006.01)
`AGIK 38/26
`(52) U.S. Ch.
`caccccccssesssecssssesteee 514/3; 514/12; 514/738
`
`(57)
`
`ABSTRACT
`
`(73)
`
`Assignee: Novo Nordisk A/S, Bagsvaerd (DK)
`
`(21)
`
`Appl. No.:
`
`11/435,977
`
`(22)
`
`Filed:
`
`May 17, 2006
`
`Related U.S. Application Data
`
`(63)
`
`Continuation of application No. PCT/DK04/00792,
`filed on Nov. 18, 2004.
`
`The present invention relates to pharmaceutical formula-
`tions comprising a peptide and propylene glycol, to methods
`of preparing such formulations, and to uses of such formu-
`lations in the treatment of diseases and conditions for which
`
`use of the peptide contained in such formulations is indi-
`cated. The present invention further relates to methods for
`reducing the clogging of injection devices by a peptide
`formulation and for reducing deposits on production equip-
`ment during production of a peptide formulation.
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 1
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 1
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 1 of 7
`
`US 2007/0010424 Al
`
`FIGURE 1
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 2
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 2
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 2 of 7
`
`US 2007/0010424 Al
`
`FIGURE 2
`
`
`
`Mannitol
`
`Argi-
`
`Inasi-
`
`Glyce-
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 3
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 3
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 3 of 7
`
`US 2007/0010424 Al
`
`FIGURE 3
`
` STE reernoeeASP LSahscReedPi
`
`Myo-inositol
`
`Maltose
`
`Glycerol
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 4
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 4
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 4 of 7
`
`US 2007/0010424 Al
`
`FIGURE 4
`
`Glycine
`
`Lactose
`
`Mannitol
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 5
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 5
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 5 of 7
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`US 2007/0010424 Al
`
`FIGURES
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 6
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 6
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 6 of 7
`
`US 2007/0010424 Al
`
`FIGURE 6
`
`
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 7
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 7
`
`
`
`Patent Application Publication Jan. 11,2007 Sheet 7 of 7
`
`US 2007/0010424 Al
`
`FIGURE 7
`
`Mannitol
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 8
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 8
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`PROPYLENE GLYCOL-CONTAINING PEPTIDE
`FORMULATIONS WHICH ARE OPTIMAL FOR
`PRODUCTION AND FOR USE IN INJECTION
`DEVICES
`
`CROSS REFERENCE TO RELATED
`APPLICATIONS
`
`[0001] This Application is a continuation of International
`Application serial no. PCT/DK2004/000792 filed Nov. 18,
`2004 and claimspriority from U.S. application Ser. No.
`60/524653 filed Nov. 24, 2003 and from Danish Application
`serial no. PA 2003 01719 filed Nov. 20, 2003.
`
`FIELD OF THE INVENTION
`
`invention relates to pharmaceutical
`[0002] The present
`formulations comprising a peptide and propylene glycol, to
`methods of preparing such formulations, and to uses of such
`formulations in the treatment of diseases and conditions for
`which use of the peptide contained in such formulations is
`indicated. The present invention further relates to methods
`for reducing the clogging of injection devices by a peptide
`formulation and for reducing deposits on production equip-
`ment during production of a peptide formulation.
`
`BACKGROUND OF THE INVENTION
`
`[0003] The inclusion of isotonicity agents in peptide-
`containing pharmaceutical formulations is widely known
`and one of the more commonisotonic agents used in such
`formulations is mannitol. However, the present inventors
`have observed that mannitol causes problems during the
`production of peptide formulationsas it crystallizes resulting
`in deposits in the production equipment and in the final
`product. Such deposits increase the need to clean thefilling
`equipment during production of the formulation and this
`results in reduced production capability. In addition, such
`deposits may also result in reducedyield ofthe final product
`since vials/cartridges containing the peptide formulation
`may needto be discardedif particles are present. Finally, the
`present inventors have observed that in peptide formulations
`to be administered by injection, the presence of mannitol
`results in clogging of injection devices.
`
`[0004] Accordingly, it is desirable to identify an alterna-
`tive isotonic agent to mannitol for inclusion in peptide-
`containing formulations and in particular, for inclusion in
`peptide formulations which are administered by injection.
`
`SUMMARY OF THE INVENTION
`
`[0005] The present inventors have discovered that peptide
`formulations containing propylene glycol at certain concen-
`trations exhibit reduced deposits in production equipment
`and in the final product and also exhibit reduced clogging of
`injection devices. The present compositions may be formu-
`lated with any peptide and are also physically and chemi-
`cally stable thus rendering them shelf-stable and suitable for
`invasive (eg. injection, subcutaneous injection, intramuscu-
`lar, intraveneous or infusion) as well as non-invasive (eg
`nasal, oral, pulmonary,
`transdermal or transmucosal e.g.
`buccal) means of administration.
`
`[0006] The present invention therefore relates to a phar-
`maceutical formulation comprising a peptide and propylene
`glycol, where the propylene glycol is present in a concen-
`tration of 1-100 mg/mland the pH of the formulation is from
`
`7-10. In a preferred embodiment, the pharmaceutical for-
`mulations of the invention further contain a buffer and a
`
`preservative.
`
`[0007] The present invention also relates to methods for
`producing the pharmaceutical formulations of the invention.
`
`In one embodiment, the method for preparing a
`[0008]
`peptide formulation comprises:
`
`a) preparing a first solution by dissolving pre-
`[0009]
`servative, propylene glycol and buffer in water;
`
`b) preparing a second solution by dissolving the
`[0010]
`peptide in water;
`
`[0011]
`
`c) mixing the first and second solutions; and
`
`d) adjusting the pH of the mixture in c) to the
`[0012]
`desired pH.
`
`In another embodiment, the method for preparing a
`[0013]
`peptide formulation comprises:
`
`a) preparing a first solution by dissolving pre-
`[0014]
`servative and buffer in water;
`
`[0015]
`
`b) adding propylene glycol to the first solution;
`
`c) mixing the first solution with a second solu-
`[0016]
`tion containing peptide dissolved in water; and
`
`d) adjusting the pH of the mixture in c) to the
`[0017]
`desired pH.
`
`In yet another embodiment, the method for prepar-
`[0018]
`ing a peptide formulation comprises:
`
`a) preparing a solution by dissolving preserva-
`[0019]
`tive, buffer and propylene glycol in water;
`
`[0020]
`and
`
`b) adding the peptide to the solution of step a);
`
`c) adjusting the pH ofthe solution of step b) to
`[0021]
`the desired pH.
`
`[0022] The present invention further relates to methods of
`treatment using the pharmaceutical
`formulations of the
`invention where the compositions are administered in an
`amounteffective to combat the disease, condition, or disor-
`der for which administration of the peptide contained in the
`formulation is indicated.
`
`In addition the present invention also relates to a
`[0023]
`method for reducing deposits on production equipment
`during production of a peptide formulation, where the
`method comprises replacing the isotonicity agent previously
`utilized in said formulation with propylene glycol at a
`concentration of between 1-100 mg/ml.
`
`In one embodiment, the reduction in deposits on
`[0024]
`the production equipment during production by the propy-
`lene glycol-containing formulation relative to that observed
`for the formulation containing the previously utilized iso-
`tonicity agent is measured by a simulatedfilling experiment.
`
`[0025] The present invention also relates to a method for
`reducing deposits in the final product during production of
`a peptide formulation, where the method comprises replac-
`ing the isotonicity agent previously utilized in said formu-
`lation with propylene glycol at a concentration of between
`1-100 mg/ml.
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 9
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 9
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`In one embodiment, the reduction in deposits in the
`[0026]
`final product is measured by a reduction in the number of
`vials and/or cartridges of the propylene glycol-containing
`formulation that must be discarded due to deposits relative
`to number of vials and/or cartridges of the formulation
`containing the previously utilized isotonicity agent that must
`be discarded due to deposits.
`
`[0027] The present invention further relates to a method
`for reducing the clogging of injection devices by a peptide
`formulation, where the method comprises replacing the
`isotonicity agent previously utilized in said formulation with
`propylene glycol at a concentration of between 1-100
`mg/ml.
`
`In one embodiment, the reduction in clogging of
`[0028]
`the injection device by the propylene glycol-containing
`formulation relative to that observed for the formulation
`containing the previously utilized isotonicity agent is mea-
`sured in a simulated in use study.
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`[0029] FIG. 1 shows a photograph of dried droplets on
`microscopeslides of from left to right, placebo (no peptide)
`formulations containing no isotonic agent (e only water,
`preservative and buffer), mannitol, sorbitol, xylitol, sucrose
`or glycerol as the isotonic agent with the far right slide
`containing mannitol with peptide Arg**, Lys*°(N*-(y-
`Glu(N@-hexadecanoyl)))-GLP-1 (7-37).
`
`[0030] FIG. 2 showslight microscopypictures of from left
`to right, some of the dried droplets of placebo formulations
`containing mannitol, arginin,
`inositol or glycerol as the
`isotonic agent.
`
`[0031] FIG. 3 shows light microscopypictures of clogged
`needles dosed with placebo formulations containing myo-
`inositol, maltose or glycerol as the isotonic agent.
`
`[0032] FIG. 4 showslight microscopy pictures of deposits
`on needles dosed with placebo formulations containing
`glycine, lactose or mannitol as the isotonic agent.
`
`[0033] FIG. 5 showsfilling equipment after 24 hours
`simulated filling with Arg**, Lys*°(N*-(y-Glu(N°-hexade-
`canoyl)))-GLP-1(7-37) medium containing myo-inositol.
`
`[0034] FIG. 6 showsdeposits onfilling equipment after 24
`hours simulated filling with a mannitol-containing placebo
`formulation.
`
`[0035] FIG. 7 shows deposits on needles dosed with
`mannitol (top panel) and propylene glycol (bottom panel)-
`containing Arg**, Lys*°(N*-(y-Glu(N°-hexadecanoyl)))-
`GLP-1(7-37) formulations.
`
`DESCRIPTION OF THE INVENTION
`
`[0036] The present invention relates to a pharmaceutical
`formulation comprising a peptide or a mixture of peptides
`and propylene glycol where the final concentration of pro-
`pylene glycol in the formulation is 1-100 mg/ml and the pH
`of the formulation is in the range of from 7-10.
`
`[0037] The pharmaceutical formulations of the invention
`are found to be optimal for production because they exhibit
`reduced deposits in production equipmentrelative to for-
`are
`agonists
`identifying GLP-1
`for
`[0042] Methods
`mulations containing other isotonicity agents as measured
`by the simulatedfilling studies described in the Examples. In
`described in WO 93/19175 (Novo Nordisk A/S) and
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 10
`
`addition, the pharmaceutical formulations of the invention
`are found to be optimal for use in injection devices because
`they exhibit reduced clogging of the injection devicesrela-
`tive to formulations containing other isotonicity agents as
`measured by the simulated in use studies described in the
`Examples.
`[0038] The formulations of the present invention may be
`formulated with any peptide where examples of such pep-
`tides include, butare notlimited to, glucagon, human growth
`hormone (hGH), insulin, aprotinin, FactorVII, tissue plas-
`minogen activator
`(TPA), FactorVIa, FFR-FactorVIa,
`heparinase, ACTH, Heparin Binding Protein, corticotropin-
`releasing factor, angio-tensin, calcitonin, glucagon-like pep-
`tide-1, glucagon-like peptide-2, insulin-like growth factor-1,
`insulin-like growth factor-2, fibroblast growth factors, gas-
`tric inhibitory peptide, growth hormone-releasing factor,
`pituitary adenylate cyclase activating peptide,
`secretin,
`enterogastrin, somatostatin, somatomedin, parathyroid hor-
`mone, thrombopoietin, erythropoietin, hypothalamic releas-
`ing factors, prolactin, thyroid stimulating hormones, endor-
`phins, enkephalins, vasopressin, oxytocin, opiods, DPP IV,
`interleukins,
`immunoglobulins,
`complement
`inhibitors,
`serine protease inhibitors, cytokines, cytokine receptors,
`PDGF, tumornecrosis factors, tumor necrosis factors recep-
`tors, growth factors and analogues as well as derivatives
`thereof where each of these peptides constitutes an alterna-
`tive embodiment of the present invention.
`[0039]
`In the present application,
`the designation “an
`analogue” is used to designate a peptide wherein one or
`more amino acid residues of the parent peptide have been
`substituted by another amino acid residue and/or wherein
`one or more aminoacid residues of the parent peptide have
`been deleted and/or wherein one or more amino acid resi-
`
`dues have been added to the parent peptide. Such addition
`can take place either at the N-terminal end or at the C-ter-
`minal end of the parent peptide or both. Typically “an
`analogue” is a peptide wherein 6 or less amino acids have
`been substituted and/or added and/or deleted from the parent
`peptide, more preferably a peptide wherein 3 or less amino
`acids have been substituted and/or added and/or deleted
`
`from the parent peptide, and most preferably, a peptide
`wherein one amino acid has been substituted and/or added
`
`and/or deleted from the parent peptide.
`[0040]
`In the present application, “a derivative” is used to
`designate a peptide or analogue thereof which is chemically
`modified by introducing an organic substituent e.g. ester,
`alkyl or lipophilic functionalities, on one or more amino acid
`residues of the peptide or analogue thereof.
`[0041]
`In one embodiment, the peptide to be included in
`the formulation of the invention is a GLP-1 agonist where “a
`GLP-1 agonist” is understood to refer to any peptide which
`fully or partially activates the human GLP-1 receptor. In a
`preferred embodiment, the “GLP-1 agonist” is any peptide
`that binds to a GLP-1 receptor, preferably with an affinity
`constant (K,,) or a potency (EC.,) of below 1 uM,e.g. below
`100 nM as measured by methods knownin theart (see e.g.
`WO 98/08871) and exhibits insulinotropic activity, where
`insulinotropic activity may be measured in vivo or in vitro
`assays known to those of ordinary skill
`in the art. For
`example,
`the GLP-1 agonist may be administered to an
`animal and the insulin concentration measured over time.
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 10
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`examples of suitable GLP-1 analogues and derivatives
`which can be used according to the present
`invention
`includes those referred to in WO 99/43705 (Novo Nordisk
`A/S), WO 99/43706 (Novo Nordisk A/S), WO 99/43707
`(Novo Nordisk A/S), WO 98/08871 (analogues with lipo-
`philic substituent) and in WO 02/46227 (analogues fused to
`serum albumin or to Fe portion of an Ig).(Novo Nordisk
`A/S), WO 99/43708 (Novo Nordisk A/S), WO 99/43341
`(Novo Nordisk A/S), WO 87/06941 (The General Hospital
`Corporation), WO 90/11296 (The General Hospital Corpo-
`ration), WO 91/11457 (Buckley et al.), WO 98/43658 (Eli
`Lilly & Co.), EP 0708179-A2 (Eli Lilly & Co.), EP
`0699686-A2 (Eli Lilly & Co.), WO 01/98331 (Eli Lilly &
`Co).
`
`In one embodiment, the GLP-1 agonist is selected
`[0043]
`from the group consisting of GLP-1(7-36)-amide, GLP-1(7-
`37), a GLP-1(7-36)-amide analogue, a GLP-1(7-37) ana-
`logue, or a derivative of any of these.
`
`In one embodiment, the GLP-1 agonistis a deriva-
`[0044]
`tive of GLP-1(7-36)-amide, GLP-1(7-37), a GLP-1(7-36)-
`amide analogue or a GLP-1(7-37) analogue, which com-
`prises a lipophilic substituent.
`
`In this embodiment of the invention, the GLP-1
`[0045]
`derivative preferably has three lipophilic substituents, more
`preferably two lipophilic substituents, and most preferably
`one lipophilic substituent attached to the parent peptide (ie
`GLP-1(7-36)-amide, GLP-1(7-37), a GLP-1(7-36)-amide
`analogue or a GLP-1(7-37) analogue), where each lipophilic
`substituent(s) preferably has 4-40 carbon atoms, more pref-
`erably 8-30 carbon atoms, even more preferably 8-25 carbon
`atoms, even more preferably 12-25 carbon atoms, and most
`preferably 14-18 carbon atoms.
`
`In one embodiment,the lipophilic substituent com-
`[0046]
`prises a partially or completely hydrogenated cyclopen-
`tanophenathrene skeleton.
`
`Inanother embodiment,the lipophilic substituent is
`[0047]
`a straight-chain or branched alkyl group.
`
`In yet another embodiment, the lipophilic substitu-
`[0048]
`ent is an acyl group ofa straight-chain or branched fatty
`acid. Preferably, the lipophilic substituent is an acyl group
`having the formula CH,(CH,),CO—, wherein n is an inte-
`ger from 4 to 38, preferably an integer from 12 to 38, and
`most preferably is CH,(CH,),,CO—, CH,(CH,),,CO—,
`CH,(CH,),<CO—, CH,(CH,),gCO—, CH;(CH,)4CO—
`and CH,(CH,),.CO—.Ina more preferred embodiment,the
`lipophilic substituent is tetradecanoyl. In a most preferred
`embodiment, the lipophilic substituent is hexadecanoy]l.
`
`can be attached to one of the following functional groups of
`an amino acid of the parent GLP-1 peptide:
`
`(a) the amino groupattachedto the alpha-carbon
`[0051]
`of the N-terminal amino acid,
`
`(b) the carboxy group attached to the alpha-
`[0052]
`carbon of the C-terminal amino acid,
`
`[0053]
`
`(c) the epsilon-amino group of any Lysresidue,
`
`(d) the carboxy group of the R group of any Asp
`[0054]
`and Glu residue,
`
`(e) the hydroxy group of the R group of any Tyr,
`[0055]
`Ser and Thr residue,
`
`(f) the amino group of the R group of any Trp,
`[0056]
`Asn, Gln, Arg, and His residue, or
`
`(g) the thiol group of the R group of any Cys
`[0057]
`residue.
`
`is
`In one embodiment, a lipophilic substituent
`[0058]
`attached to the carboxy group of the R group of any Asp and
`Glu residue.
`
`In another embodiment, a lipophilic substituent is
`[0059]
`attached to the carboxy group attached to the alpha-carbon
`of the C-terminal amino acid.
`
`Ina most preferred embodiment, a lipophilic sub-
`[0060]
`stituent is attached to the epsilon-amino group of any Lys
`residue.
`
`In a preferred embodiment of the invention, the
`[0061]
`lipophilic substituentis attached to the parent GLP-1 peptide
`by means of a spacer. A spacer must contain at least two
`functional groups, oneto attach to a functional group of the
`lipophilic substituent and the other to a functional group of
`the parent GLP-1 peptide.
`
`In one embodiment, the spacer is an amino acid
`[0062]
`residue except Cys or Met, or a dipeptide such as Gly-Lys.
`For purposes of the present invention, the phrase “a dipep-
`tide such as Gly-Lys” means any combination of two amino
`acids except Cys or Met, preferably a dipeptide wherein the
`C-terminal amino acid residue is Lys, His or Trp, preferably
`Lys, and the N-terminal aminoacid residue is Ala, Arg, Asp,
`Asn, Gly, Glu, Gln, Ile, Leu, Val, Phe, Pro, Ser, Tyr, Thr, Lys,
`His and Trp. Preferably, an amino group of the parent
`peptide forms an amide bond with a carboxylic group of the
`amino acid residue or dipeptide spacer, and an amino group
`of the amino acid residue or dipeptide spacer forms an amide
`bond with a carboxyl group of the lipophilic substituent.
`
`Ina further embodimentof the present invention,
`[0049]
`the lipophilic substituent has a group which is negatively
`charged such as a carboxylic acid group. For example, the
`lipophilic substituent may be an acyl group of a straight-
`chain or branched alkane a,w-dicarboxylic acid of the
`formula HOOC(CH,),,,CO—, wherein m is an integer from
`4 to 38, preferably an integer from 12 to 38, and most
`preferably is HOOC(CH,),,CO—, HOOC(CH,), ,CO—,
`HOOC(CH,), ,CO—,
`HOOC(CH,),,CO—
`or
`HOOC(CH,),,CO—.
`
`[0063] Preferred spacers are lysyl, glutamyl, asparagyl,
`glycyl, beta-alanyl and gamma-aminobutanoyl, each of
`which constitutes an individual embodiment. Mostpreferred
`spacers are glutamyl and beta-alanyl. When the spacer is
`Lys, Glu or Asp, the carboxyl group thereof may form an
`amide bond with an amino group of the amino acid residue,
`and the amino group thereof may form an amide bond with
`a carboxyl] group of the lipophilic substituent. When Lys is
`used as the spacer, a further spacer may in someinstances be
`inserted between the e-amino group of Lys and the lipophilic
`substituent. In one embodiment, such a further spacer is
`succinic acid which forms an amide bond with the e-amino
`group of Lys and with an amino group present
`in the
`the
`In the GLP-1 derivatives of the invention,
`[0050]
`lipophilic substituent. In another embodiment such a further
`lipophilic substituent(s) contain a functional group which
`spacer is Glu or Asp which forms an amide bond with the
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 11
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 11
`
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`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`group of Lys and another amide bond with a
`€-amino
`carboxyl group present in the lipophilic substituent, thatis,
`the lipophilic substituent is a N‘-acylated lysine residue.
`
`an
`is
`spacer
`the
`embodiment,
`In another
`[0064]
`unbranched alkane a,m-dicarboxylic acid group having
`from 1 to 7 methylene groups, which spacer forms a bridge
`between an amino group ofthe parent peptide and an amino
`group of the lipophilic substituent. Preferably, the spacer is
`succinic acid.
`
`Ina further embodiment, the lipophilic substituent
`[0065]
`with the attached spacer
`is a group of the formula
`CH;(CH,),NH—CO(CH,),CO—, wherein p is an integer
`from 8 to 33, preferably from 12 to 28 and q is an integer
`from 1 to 6, preferably 2.
`
`Ina further embodiment, the lipophilic substituent
`[0068]
`is a group of the formula COOH(CH,),CO— wherein t is an
`integer from 6 to 24.
`
`Ina further embodiment, the lipophilic substituent
`[0071]
`with the attached spacer
`is a group of the formula
`—NHCH(COOH)(CH;),NH—
`COCH((CH,),COOH)NH—CO(CH,),.CH;, wherein w is
`an integer from 10 to 16.
`
`Ina further embodiment, the lipophilic substituent
`[0072]
`with the attached spacer
`is a group of the formula
`—NHCH(COOH)(CH,),NH—
`CO(CH,),CH(COOH)NHCO(CH,),.CH;, wherein x is zero
`or an integer from 1 to 22, preferably 10 to 16.
`
`In yet another embodiment the GLP-1 agonist is
`[0073]
`Arp**, Lys?°(N*-(y-Glu(N“-hexade-canoyl)))-GLP-1(7-37).
`
`In yet another embodiment the GLP-1 agonist is
`[0074]
`selected from the group consisting of Gly*-GLP-1(7-36)-
`amide, Gly*-GLP-1(7-37), Val®-GLP-1(7-36)-amide, Val?-
`GLP-1(7-37), Val®Asp?*-GLP-1(7-36)-amide, Val*Asp7?-
`GLP-1(7-37), Val®Glu??-GLP-1(7-36)-amide, Val®Glu??-
`GLP-1(7-37), Val®Lys?*-GLP-1(7-36)-amide, Val®Lys7?-
`GLP-1(7-37), ValArg?*-GLP-1(7-36)-amide, Val®Arg??-
`GLP-1(7-37), Val®His”*-GLP-1(7-36)-amide, Val*His??-
`GLP-1(7-37), analogues thereof and derivatives of any of
`these.
`
`GLP-1(7-37); Arg?°**-GLP-1(7-37); Arg*°**Lys*°-GLP-
`1(7-37),; Arg?*Lys*°-GLP-1(7-37);
` Arg**Lys*°-GLP-1(7-
`37);
`Val*Arg’?-GLP-1 (7-37);
`Met*Arg??-GLP-1(7-
`37);Gly*His??-GLP-1(7-37);
`Val*His??-GLP-1(7-37);
`Met*His”*-GLP-1(7-37); His*’-GLP-1(7-37); Gly®-GLP-
`1(7-37); Val®-GLP-1 (7-37); Met®-GLP-1(7-37); Gly’Asp??-
`GLP-1(7-37); Val?Asp??-GLP-1(7-37); Met®Asp??-GLP-
`1(7-37); Gly8Glu?*-GLP-1(7-37);_ Val®Glu?*-GLP-1(7-37);
`Met*Glu??-GLP-1(7-37);
`Gly®Lys??-GLP-1(7-37);
`Val®Lys*?-GLP-1(7-37);
`Met®Lys??-GLP-1(7-37);
`Gly*Arg??-GLP-1(7-37);
`Val®Lys”?His*’-GLP-1(7-37);
`Gly*Glu”His*’-GLP-1(7-37);
` Val®Glu*?His*’-GLP-1(7-
`37); Met®Glu??His*’-GLP-1(7-37); Gly*Lys*? His*’-GLP-
`1(7-37); Met*Lys??His*’-GLP-1(7-37);
` Gly*Arg??His*’-
`GLP-1(7-37);
`Val®Arg”*His*’-GLP-1(7-37);
`Met*Arg?*His*’-GLP-1(7-37);
` Gly®His?*His*’-GLP-1(7-
`Ina further embodiment, the lipophilic substituent
`[0066]
`37); Val$His??His*’-~GLP-1(7-37); Met*His**His *’-GLP-
`with the attached spacer
`is a group of the formula
`1(7-37); Gly*His*’-GLP-1(7-37); Val®His*’-GLP-1(7-37);
`CH;(CH,),CO—NHCH(COOH)(CH,),CO—, whereinris
`Met*His*’-GLP-1(7-37);
` Gly*Asp”?His*’-GLP-1(7-37);
`an integer from 4 to 24, preferably from 10 to 24.
`Val®Asp??His*’-GLP-1(7-37);|Met®Asp??His*’-GLP-1(7-
`37); Arg?°-GLP-1(7-36)-amide; Arg**-GLP-1(7-36)-amide;
`[0067]
`Ina further embodiment, the lipophilic substituent
`Lys*°-GLP-1(7-36)-amide;
`Arp?®*4Lys°°-GLP-1(7-36)-
`with the attached spacer
`is a group of the formula
`amide; Arg?®?4-GLP-1(7-36)-amide; Arg?°**Lys*°-GLP-
`CH;(CH,),CO—NHCH((CH,),COOH)CO—, wherein s is
`1(7-36)-amide;
`Arp?*Lys*°-GLP-1(7-36)-amide;
`an integer from 4 to 24, preferably from 10 to 24.
`Arg*“Lys?°-GLP-1(7-36)-amide; Gly*-GLP-1(7-36)-amide;
`Val®-GLP-1(7-36)-amide;
`Met®-GLP-1(7-36)-amide;
`Gly*Asp*?-GLP-1(7-36)-amide; Gly*Glu??His*’-GLP-1(7-
`36)-amide; Val®Asp??-GLP-1(7-36)-amide; Met®Asp??-
`GLP-1(7-36)-amide;
`Gly*Glu*?-GLP-1(7-36)-amide;
`Ina further embodiment, the lipophilic substituent
`[0069]
`Val®Glu??-GLP-1(7-36)-amide;©Met®Glu??-GLP-1(7-36)-
`with the attached spacer
`is a group of the formula
`amide; Gly*Lys??-GLP-1(7-36)-amide; Val*Lys*?-GLP-1(7-
`—NHCH(COOH)(CH,),NH—CO(CH,),,CH;, wherein u is
`36)-amide;
`Met®Lys?*-GLP-1(7-36)-amide;
`an integer from 8 to 18.
`Gly*His*?His*’-GLP-1(7-36)-amide; Gly*Arg*?-GLP-1(7-
`Ina further embodiment, the lipophilic substituent
`[0070]
`36)-amide; Val*Arg??-GLP-1(7-36)-amide; Met®Arg7?-
`with the attached spacer
`is a group of the formula
`GLP-1(7-36)-amide;
`Gly*His??-GLP-1(7-36)-amide;
`CH,(CH,),CO—NH—(CH,),—_CO,whereinvis an integer
`Val*His*?-GLP-1(7-36)-amide;|Met®His*?~-GLP-1(7-36)-
`from 4 to 24 and z is an integer from 1 to 6.
`amide; His*’-GLP-1(7-36)-amide,; Val*Arg?*His*’-GLP-
`1(7-36)-amide; Met*Arg*’-GLP-1(7-36)-amide; Gly*His*’-
`GLP-1(7-36)-amide;
`Val®His*’-GLP-1(7-36)-amide;
`Met*His*’-GLP-1(7-36)-amide; Gly*Asp?? His*’-GLP-1(7-
`36)-amide;
`Val®Asp??His*’-~GLP-1(7-36)-amide;
`Met*Asp?*His*’-GLP-1(7-36)-amide;
`Val®Glu?*His??-
`GLP-1(7-36)-amide; Met®Glu??His*’-GLP-1(7-36)-amide;
`Gly*Lys”*His*’-GLP-1(7-36)-amide; Val*Lys**His*’-GLP-
`1(7-36)-amide;
`Met®Lys?*His*’-GLP-1(7-36)-amide;
`Gly*Arg?*His*’-GLP-1(7-36)-amide; Val*His**His*’-GLP-
`1(7-36)-amide; Met*His?*His*’-GLP-1(7-36)-amide; and
`derivatives thereof.
`
`In yet another embodiment the GLP-1 agonist is
`[0076]
`selected from the group consisting of Val®Trp'*Glu??-GLP-
`1(7-37),
` Val®Glu?*Val*°-GLP-1(7-37),
` Val®Tyr'®Glu?-
`GLP-1(7-37),
`Val®Trp'°Glu?-GLP-1(7-37),
`Val®Leu!®Glu*?-GLP-1(7-37),
` Val®Tyr'8Glu??-GLP-1(7-
`37), Val®Glu?? His*’-GLP-1(7-37), Val®Glu?7Ile*?-GLP-
`1(7-37),
`Val®Trp! ®Glu??Val?*Te*?-GLP-1(7-37),
`Val®Trp! °Glu?7Ie*? -GLP-1(7-37),
`Val®Glu??Val?*Ie*?-
`GLP-1(7-37), Val°Trp'°Glu**Val?>-GLP-1(7-37), analogues
`thereof and derivatives of any of these.
`
`In yet another embodiment the GLP-1 agonist is
`[0077]
`exendin-4 or exendin-3, an exendin-4 or exendin-3 analogue
`or a derivative of any of these.
`
`In yet another embodiment the GLP-1 agonist is
`[0075]
`selected from the group consisting of Arg?°-GLP-1(7-37);
`[0078] Examples of exendins as well as analogues, deriva-
`
`Arg**-GLP-1(7-37);|Lys*°-GLP-1(7-37); Arg?®?*Lys*°- tives, and fragments thereof to be included within the
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 12
`
`SUN EXHIBIT 1016, IPR2024-00107, PAGE 12
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`present invention are those disclosed in WO 97/46584, U.S.
`Pat. No. 5,424,286 and WO 01/04156. U.S. Pat. No. 5,424,
`286 describes a method for stimulating insulin release with
`an exendin polypeptide. The exendin polypeptides disclosed
`include HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGX;
`wherein
`X=P
`or
`Y,
`and
`HX1X2GTFITSDLSKQMEEEAVRLFIEWLKNGGPSSG
`APPPS; wherein X1X2=SD (exendin-3) or GE (exendin-4)).
`WO 97/46584 describes truncated versions of exendin pep-
`tide(s). The disclosed peptides increase secretion and bio-
`synthesis of insulin, but reduce those of glucagon. WO
`01/04156 describes exendin-4 analogues and derivatives as
`well as the preparation of these molecules. Exendin-4 ana-
`logues stabilized by fusion to serum albumin or Fe portion
`of an Ig are disclosed in WO 02/46227.
`
`the exendin-4 analogue is
`In one embodiment,
`[0079]
`HGEGTFTSDLSKQMEEEAVR-
`LFIEWLKNGGPSSGAPPSKKKKKK-amide.
`
`[0080] Where the peptide to be includedin the formulation
`of the invention is a GLP-1 agonist, the GLP-1 agonist is
`present in a concentration from about 0.1 mg/ml to about
`100 mg/ml, more preferably in a concentration from about
`0.1 mg/ml to about 50 mg/ml, and most preferably in a
`concentration of from about 0.1 mg/ml to about 10 mg/ml.
`
`In another embodiment, the peptide to be included
`[0081]
`in the formulation of the invention is insulin, where “insu-
`lin” is understood to mean humaninsulin, [where “human
`insulin” means insulin having the amino acid sequence
`shown in DSHW Nicol and L F Smith: Nature,
`(1960)
`4736:483-485, which is hereby incorporated by reference],
`human insulin analogs, human insulin derivatives or mix-
`tures thereof, where examples of insulin analogs and deriva-
`tives are those disclosed in EP 0 792 290 (Novo Nordisk
`A/S), EP 0 214 826 and EP 0 705 275 (Novo Nordisk A/S),
`US. Pat. No. 5,504,188 (Eli Lilly), EP 0 368 187 (Aventis),
`US. Pat. Nos. 5,750,497 and 6,011,007, EP 375437 and EP
`383472 and where such insulins may include, but are not
`limited to, NPH insulin, Lys 629 (Ne-tetradecanoyl)
`des(B30) human insulin, Lys®*?-(N*-(y-glutamyl-N“-litho-
`cholyl) des(B30) human insulin, N°??°-octanoyl
`insulin,
`30/70 mixtures of prompt insulin zinc (SemiLente®) with
`extended insulin zinc (Ultralente®), sold commercially as
`Lente®, insulin glargine (Lantus®) or extended insulin zinc
`(Ultralente®), Lys®?* Pro??? human insulin (Huma-log®),
`Asp
`human insulin, insulin aspart (Novolog®), or a 30/70
`mixture of insulin aspart and insulin aspart protamine
`(NovoMix®).
`
`In one embodiment, the insulin is a derivative of
`[0082]
`human insulin or a human insulin analogue where the
`derivative contains at least one lysine residue and a lipo-
`philic substituent is attached to the epsilon amino group of
`the lysine residue.
`
`In one embodiment,the lysine residue to which the
`[0083]
`lipophilic substituent is attachedis present at position B28 of
`the insulin peptide.
`
`In an alternative embodiment, the lysine residue to
`[0084]
`which the lipophilic substituent is attached is present at
`position B29 of the insulin peptide.
`
`In yet another embodiment, lipophilic substituent is
`[0085]
`an acyl group corresponding to a carboxylic acid having at
`least 6 carbon atoms.
`
`the lipophilic
`In another preferred embodiment,
`[0086]
`substituent is an acyl group, branched or unbranched, which
`corresponds to a carboxylic acid having a chain of carbon
`atoms 8 to 24 atomslong.
`
`the lipophilic
`In another preferred embodiment,
`[0087]
`substituent is an acyl group corresponding to a fatty acid
`having at least 6 carbon atoms.
`
`the lipophilic
`In another preferred embodiment,
`[0088]
`substituent
`is an acyl group corresponding to a linear,
`saturated carboxylic acid having from 6 to 24 carbon atoms.
`
`the lipophilic
`In another preferred embodiment,
`[0089]
`substituent
`is an acyl group corresponding to a linear,
`saturated carboxylic acid having from 8 to 12 carbon atoms.
`
`the lipophilic
`In another preferred embodiment,
`[0090]
`substituent
`is an acyl group corresponding to a linear,
`saturated carboxylic acid having from 10 to 16 carbon
`atoms.
`
`the lipophilic
`In another preferred embodiment,
`[0091]
`substituent is an oligo oxyethylene group comprising up to
`10, preferably up to 5, oxyethylene units.
`
`the lipophilic
`In another preferred embodiment,
`[0092]
`substituent is an oligo oxypropylene group comprising up to
`10, preferably up to 5, oxypropylene units.
`
`In one preferred embodiment, the invention relates
`[0093]
`to a human insulin derivative in which the B30 amino acid
`residue is deleted or is any amino acid residue which can be
`coded for by the genetic code except Lys, Arg and Cys; the
`A21 and the B3 aminoacid residues are, independently, any
`amino acid residues which can be coded for by the genetic
`
`code except Lys, Arg and Cys; Phe?’ may be deleted; the
`
`
`
`-amino group of Lys®”?hasa lipophilic substituent which
`comprisesat least 6 carbon atoms; and 2-4 Zn** ions may be
`boundto each insulin hexamer with