`(12) Patent Application Publication (10) Pub. No.: US 2007/0010424 A1
`(43) Pub. Date:
`Jan. 11, 2007
`Pedersen et al.
`
`US 200700 10424A1
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`(54)
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`PROPYLENE GLYCOL-CONTAINING
`PEPTIDE FORMULATIONS WHICH ARE
`OPTIMIAL FOR PRODUCTION AND FOR
`USE IN INUECTION DEVICES
`
`Inventors: Tina Bjeldskov Pedersen, Smorum
`(DK); Claude Bonde, Lyngby (DK);
`Dorthe Kot Engelund, Holte (DK)
`Correspondence Address:
`NOVO NORDISK, INC.
`PATENT DEPARTMENT
`1OO COLLEGE ROAD WEST
`PRINCETON, NJ 08540 (US)
`Assignee: Novo Nordisk A/S, Bagsvaerd (DK)
`Appl. No.:
`11/435,977
`
`Filed:
`
`May 17, 2006
`
`Related U.S. Application Data
`Continuation of application No. PCT/DK04/00792,
`filed on Nov. 18, 2004.
`
`(30)
`
`Foreign Application Priority Data
`
`Nov. 20, 2003 (DK)................................ PA 2003 O1719
`
`Publication Classification
`
`(51) Int. Cl.
`(2006.01)
`A6II 38/28
`(2006.01)
`A6II 3L/045
`(2006.01)
`A6II 38/26
`(52) U.S. Cl. ................................. 514/3: 514/12: 514/738
`
`(57)
`
`ABSTRACT
`
`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.
`
`MPI EXHIBIT 1016 PAGE 1
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`MPI EXHIBIT 1016 PAGE 1
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`Patent Application Publication Jan. 11, 2007 Sheet 1 of 7
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`US 2007/0010424 A1
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`FIGURE 1
`
`
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`MPI EXHIBIT 1016 PAGE 2
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`MPI EXHIBIT 1016 PAGE 2
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`
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`Patent Application Publication Jan. 11, 2007 Sheet 2 of 7
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`US 2007/0010424 A1
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`
`
`FIGURE 2
`
`Mannitol
`
`Argi-
`
`Glyce
`
`MPI EXHIBIT 1016 PAGE 3
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`MPI EXHIBIT 1016 PAGE 3
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`
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`Patent Application Publication Jan. 11, 2007 Sheet 3 of 7
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`US 2007/0010424 A1
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`FIGURE 3
`
`
`
`SY
`
`stres a 1.
`
`s
`
`Garra is stics after it
`
`Myo-inositol
`
`Matose
`
`Glycerol
`
`MPI EXHIBIT 1016 PAGE 4
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`MPI EXHIBIT 1016 PAGE 4
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`
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`Patent Application Publication Jan. 11, 2007 Sheet 4 of 7
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`US 2007/0010424 A1
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`FGURE 4
`
`
`
`
`
`an againstreets or
`
`restsea
`
`Sct
`
`Glycine
`
`Lactose
`
`Mannito
`
`MPI EXHIBIT 1016 PAGE 5
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`MPI EXHIBIT 1016 PAGE 5
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`
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`Patent Application Publication Jan. 11, 2007 Sheet 5 of 7
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`US 2007/0010424 A1
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`FIGURE 5
`
`
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`MPI EXHIBIT 1016 PAGE 6
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`MPI EXHIBIT 1016 PAGE 6
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`
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`Patent Application Publication Jan. 11, 2007 Sheet 6 of 7
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`US 2007/0010424 A1
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`FIGURES
`
`
`
`MPI EXHIBIT 1016 PAGE 7
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`MPI EXHIBIT 1016 PAGE 7
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`
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`Patent Application Publication Jan. 11, 2007 Sheet 7 of 7
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`US 2007/0010424 A1
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`FIGURE 7
`
`
`
`Manic
`
`MPI EXHIBIT 1016 PAGE 8
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`MPI EXHIBIT 1016 PAGE 8
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`US 2007/00 10424 A1
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`Jan. 11, 2007
`
`PROPYLENE GLYCOL-CONTAINING PEPTIDE
`FORMULATIONS WHICHARE OPTIMIAL FOR
`PRODUCTION AND FOR USE IN INUECTION
`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 claims priority 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
`0002 The present invention relates to pharmaceutical
`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 common isotonic agents used in such
`formulations is mannitol. However, the present inventors
`have observed that mannitol causes problems during the
`production of peptide formulations as it crystallizes resulting
`in deposits in the production equipment and in the final
`product. Such deposits increase the need to clean the filling
`equipment during production of the formulation and this
`results in reduced production capability. In addition, Such
`deposits may also result in reduced yield of the final product
`since vials/cartridges containing the peptide formulation
`may need to be discarded if 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/ml and 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.
`0008. In one embodiment, the method for preparing a
`peptide formulation comprises:
`0009 a) preparing a first solution by dissolving pre
`servative, propylene glycol and buffer in water,
`0010 b) preparing a second solution by dissolving the
`peptide in water,
`c) mixing the first and second solutions; and
`0.011)
`0012 d) adjusting the pH of the mixture in c) to the
`desired pH.
`0013 In another embodiment, the method for preparing a
`peptide formulation comprises:
`0014) a) preparing a first solution by dissolving pre
`servative and buffer in water;
`0015 b) adding propylene glycol to the first solution;
`0016 c) mixing the first solution with a second solu
`tion containing peptide dissolved in water; and
`0017 d) adjusting the pH of the mixture in c) to the
`desired pH.
`0018. In yet another embodiment, the method for prepar
`ing a peptide formulation comprises:
`0019 a) preparing a solution by dissolving preserva
`tive, buffer and propylene glycol in water,
`0020 b) adding the peptide to the solution of step a);
`and
`0021 c) adjusting the pH of the solution of step b) to
`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
`amount effective to combat the disease, condition, or disor
`der for which administration of the peptide contained in the
`formulation is indicated.
`0023. In addition the present invention also relates to a
`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.
`0024. In one embodiment, the reduction in deposits on
`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 simulated filling 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.
`
`MPI EXHIBIT 1016 PAGE 9
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`
`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.
`0028. In one embodiment, the reduction in clogging of
`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
`microscope slides 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 shows light microscopy pictures 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 microscopy pictures of clogged
`needles dosed with placebo formulations containing myo
`inositol, maltose or glycerol as the isotonic agent.
`0032 FIG. 4 shows light microscopy pictures of deposits
`on needles dosed with placebo formulations containing
`glycine, lactose or mannitol as the isotonic agent.
`0033 FIG. 5 shows filling 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 shows deposits on filling 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-GlucN'-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 equipment relative to for
`mulations containing other isotonicity agents as measured
`by the simulated filling studies described in the Examples. In
`
`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 devices rela
`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, but are not limited to, glucagon, human growth
`hormone (hGH), insulin, aprotinin, FactorVII, tissue plas
`minogen activator (TPA), FactorVIIa, FFR-FactorVIIa,
`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, tumor necrosis 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 amino acid 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 (Ki) or a potency (ECso) of below 1 LM, e.g. below
`100 nM as measured by methods known in the art (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.
`0042 Methods for identifying GLP-1 agonists are
`described in WO 93/19175 (Novo Nordisk A/S) and
`
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`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 Fc 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
`06996.86-A2 (Eli Lilly & Co.), WO 01/98331 (Eli Lilly &
`Co).
`0043. In one embodiment, the GLP-1 agonist is selected
`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.
`0044) In one embodiment, the GLP-1 agonist is a deriva
`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.
`0045. In this embodiment of the invention, the GLP-1
`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.
`0046.
`In one embodiment, the lipophilic substituent com
`prises a partially or completely hydrogenated cyclopen
`tanophenathrene skeleton.
`0047. In another embodiment, the lipophilic substituent is
`a straight-chain or branched alkyl group.
`0.048. In yet another embodiment, the lipophilic substitu
`ent is an acyl group of a straight-chain or branched fatty
`acid. Preferably, the lipophilic substituent is an acyl group
`having the formula CH-(CH2)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-(CH2)CO—, CH (CH2)CO—, CH (CH2)CO—
`and CH (CH) CO . In a more preferred embodiment, the
`lipophilic substituent is tetradecanoyl. In a most preferred
`embodiment, the lipophilic Substituent is hexadecanoyl.
`0049. In a further embodiment of the present invention,
`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 C.()-dicarboxylic acid of the
`formula HOOC(CH2)CO—, wherein m is an integer from
`4 to 38, preferably an integer from 12 to 38, and most
`preferably is HOOC(CH2)CO-, HOOC(CH2)CO ,
`HOOC(CH2)9CO ,
`HOOC(CH2)CO-
`O
`HOOC(CH2)CO-.
`0050. In the GLP-1 derivatives of the invention, the
`lipophilic Substituent(s) contain a functional group which
`
`can be attached to one of the following functional groups of
`an amino acid of the parent GLP-1 peptide:
`0051
`(a) the amino group attached to the alpha-carbon
`of the N-terminal amino acid,
`0.052
`(b) the carboxy group attached to the alpha
`carbon of the C-terminal amino acid,
`0053 (c) the epsilon-amino group of any Lys residue,
`0054 (d) the carboxy group of the R group of any Asp
`and Glu residue,
`0055 (e) the hydroxy group of the R group of any Tyr,
`Ser and Thr residue,
`0056 (f) the amino group of the R group of any Trp,
`ASn, Gln, Arg, and His residue, or
`0057 (g) the thiol group of the R group of any Cys
`residue.
`0058. In one embodiment, a lipophilic substituent is
`attached to the carboxy group of the R group of any Asp and
`Glu residue.
`0059. In another embodiment, a lipophilic substituent is
`attached to the carboxy group attached to the alpha-carbon
`of the C-terminal amino acid.
`0060. In a most preferred embodiment, a lipophilic sub
`stituent is attached to the epsilon-amino group of any Lys
`residue.
`0061. In a preferred embodiment of the invention, the
`lipophilic substituent is attached to the parent GLP-1 peptide
`by means of a spacer. A spacer must contain at least two
`functional groups, one to attach to a functional group of the
`lipophilic Substituent and the other to a functional group of
`the parent GLP-1 peptide.
`0062. In one embodiment, the spacer is an amino acid
`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 amino acid 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.
`0063 Preferred spacers are lysyl, glutamyl, asparagyl,
`glycyl, beta-alanyl and gamma-aminobutanoyl, each of
`which constitutes an individual embodiment. Most preferred
`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 some instances 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
`lipophilic substituent. In another embodiment such a further
`spacer is Glu or Asp which forms an amide bond with the
`
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`e-amino group of Lys and another amide bond with a
`carboxyl group present in the lipophilic Substituent, that is,
`the lipophilic substituent is a N-acylated lysine residue.
`0064. In another embodiment, the spacer is an
`unbranched alkane C,c)-dicarboxylic acid group having
`from 1 to 7 methylene groups, which spacer forms a bridge
`between an amino group of the parent peptide and an amino
`group of the lipophilic substituent. Preferably, the spacer is
`Succinic acid.
`0065. In a further embodiment, the lipophilic substituent
`with the attached spacer is a group of the formula
`CH(CH), NH CO(CH2)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.
`0066.
`In a further embodiment, the lipophilic substituent
`with the attached spacer is a group of the formula
`CH(CH),CO. NHCH(COOH)(CH)CO , wherein r is
`an integer from 4 to 24, preferably from 10 to 24.
`0067. In a further embodiment, the lipophilic substituent
`with the attached spacer is a group of the formula
`CH(CH2)CO. NHCH((CH2)COOH)CO , wherein s is
`an integer from 4 to 24, preferably from 10 to 24.
`0068. In a further embodiment, the lipophilic substituent
`is a group of the formula COOH(CH),CO— whereint is an
`integer from 6 to 24.
`0069. In a further embodiment, the lipophilic substituent
`with the attached spacer is a group of the formula
`-NHCH(COOH)(CH)NH CO(CH), CH, wherein u is
`an integer from 8 to 18.
`0070. In a further embodiment, the lipophilic substituent
`with the attached spacer is a group of the formula
`CH(CH2)CO. NH-(CH) CO, wherein V is an integer
`from 4 to 24 and Z is an integer from 1 to 6.
`0071. In a further embodiment, the lipophilic substituent
`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.
`0072. In a further embodiment, the lipophilic substituent
`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.
`0073. In yet another embodiment the GLP-1 agonist is
`Arg, Lys'(N-(y-GlucN'-hexade-canoyl)))-GLP-1 (7-37).
`0074. In yet another embodiment the GLP-1 agonist is
`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 Asp
`GLP-1 (7-37), Val Glu-GLP-1 (7-36)-amide, Val Glu
`GLP-1 (7-37), Valys’-GLP-1 (7-36)-amide, Val Lys’-
`GLP-1 (7-37), ValArg-GLP-1 (7-36)-amide, ValArg
`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.
`0075. In yet another embodiment the GLP-1 agonist is
`selected from the group consisting of Arg-GLP-1 (7-37):
`Arg-GLP-1 (7-37); Lys-GLP-1 (7-37); Arg-Lys
`
`GLP-1(7-37); Arg-GLP-1 (7-37); Arg-Lys-GLP
`1(7-37); ArgLys-GLP-1 (7-37); Arg'Lys-GLP-1 (7-
`37):
`ValArg’-GLP-1 (7-37):
`MetArg’-GLP-1 (7-
`37):Gly'His?-GLP-1 (7-37):
`Val His??-GLP-1 (7-37):
`MetHis’-GLP-1 (7-37); His 7-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); MetAsp?-GLP
`1(7-37); Gly'Glu’-GLP-1(7-37); Val Glu-GLP-1 (7-37);
`MetGlu-GLP-1(7-37):
`GlyLys?-GLP-1 (7-37):
`Val Lys’-GLP-1(7-37);
`MetLys’-GLP-1 (7-37):
`GlyArg-GLP-1 (7-37):
`Val Lys?His7-GLP-1(7-37):
`GlyGlu’His7-GLP-1 (7-37);
`Val Clu’His7-GLP-1 (7-
`37); MetGlu-His7-GLP-1(7-37); GlyLys? His 7-GLP
`1(7-37); MetLys’His7-GLP-1 (7-37); GlyArg’His7
`GLP-1(7-37);
`ValArg’His7-GLP-1 (7-37):
`MetArg’His7-GLP-1 (7-37); GlyHis His 7-GLP-1 (7-
`37); Val His’His7-GLP-1 (7-37); MetHis’His 7-GLP
`1(7-37); GlyHis7-GLP-1(7-37); Val His7-GLP-1 (7-37);
`MetHis7-GLP-1 (7-37);
`GlyAsp?His7-GLP-1(7-37):
`Val Asp?His7-GLP-1(7-37); Met Asp?His 7-GLP-1 (7-
`37); Arg-GLP-1 (7-36)-amide: Arg-GLP-1 (7-36)-amide:
`Lys-GLP-1 (7-36)-amide:
`Arg-Lys-GLP-1 (7-36)-
`amide: Arg-GLP-1 (7-36)-amide: Arg-Lys-GLP
`1 (7-36)-amide:
`Arg'Lys-GLP-1 (7-36)-amide:
`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:
`GlyAsp’-GLP-1 (7-36)-amide: GlyGlu-His-GLP-1 (7-
`36)-amide: Val Asp’-GLP-1 (7-36)-amide: Met'Asp
`GLP-1 (7-36)-amide:
`Glyglu’-GLP-1 (7-36)-amide:
`ValGlu-GLP-1 (7-36)-amide: MetGlu-GLP-1 (7-36)-
`amide: GlyLys’-GLP-1 (7-36)-amide: Val Lys’-GLP-1 (7-
`36)-amide:
`MetLys’-GLP-1 (7-36)-amide:
`GlyHis’His7-GLP-1 (7-36)-amide: GlyArg-GLP-1 (7-
`36)-amide: Val Arg-GLP-1 (7-36)-amide: MetArg
`GLP-1 (7-36)-amide:
`GlyHis’-GLP-1 (7-36)-amide:
`ValHis’-GLP-1 (7-36)-amide;
`MetHis’-GLP-1 (7-36)-
`amide: His 7-GLP-1 (7-36)-amide: ValArg’His 7-GLP
`1(7-36)-amide: MetArg-GLP-1 (7-36)-amide: GlyHis 7
`GLP-1 (7-36)-amide:
`Val His 7-GLP-1 (7-36)-amide:
`MetHis7-GLP-1 (7-36)-amide: Gly-Asp’ His 7-GLP-1 (7-
`36)-amide:
`Val Asp?His 7-GLP-1 (7-36)-amide:
`Met Asp?His 7-GLP-1 (7-36)-amide:
`ValGlu22His 7
`GLP-1 (7-36)-amide; MetClu’His-GLP-1 (7-36)-amide:
`GlyLys’His7-GLP-1 (7-36)-amide: Valys’His7-GLP
`1 (7-36)-amide:
`MetLys’ His 7-GLP-1 (7-36)-amide:
`GlyArg’His 7-GLP-1 (7-36)-amide: Val His’His7-GLP
`1(7-36)-amide: MetHis His 7-GLP-1 (7-36)-amide; and
`derivatives thereof.
`0076.
`In yet another embodiment the GLP-1 agonist is
`selected from the group consisting of Val Trp'Gluff-GLP
`1(7-37), Val Glu2Val25-GLP-1 (7-37), ValTyrlGlu22
`GLP-1(7-37),
`ValTrp'Glu’-GLP-1 (7-37),
`Val Leu'Glu-GLP-1 (7-37), ValTyr'Glu-GLP-1 (7-
`37), Val Glu’ His 7-GLP-1 (7-37), Val Glu’Ile-GLP
`1(7-37),
`Val TrpGlu-Val-Ile-GLP-1 (7-37),
`Val TrpGlu-Ile-GLP-1(7-37),
`Val Glu22Val?Ile
`GLP-1 (7-37), ValTrp'Glu’Val-GLP-1 (7-37), analogues
`thereof and derivatives of any of these.
`0077. In yet another embodiment the GLP-1 agonist is
`exendin-4 or exendin-3, an exendin-4 or exendin-3 analogue
`or a derivative of any of these.
`0078 Examples of exendins as well as analogues, deriva
`tives, and fragments thereof to be included within the
`
`MPI EXHIBIT 1016 PAGE 12
`
`MPI EXHIBIT 1016 PAGE 12
`
`
`
`US 2007/00 10424 A1
`
`Jan. 11, 2007
`
`present invention are those disclosed in WO97746584, 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
`O
`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 Fc portion
`of an Ig are disclosed in WO 02/46227.
`0079. In one embodiment, the exendin-4 analogue is
`HGEGTFTSDLSKQMEEEAVR
`LFIEWLKNGGPSSGAPPSKKKKKK-amide.
`0080 Where the peptide to be included in 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.
`0081. In another embodiment, the peptide to be included
`in the formulation of the invention is insulin, where “insu
`lin' is understood to mean human insulin, 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), EPO 214826 and EP 0705 275 (Novo Nordisk A/S),
`U.S. Pat. No. 5,504,188 (Eli Lilly), EP 0368 187 (Aventis),
`U.S. Pat. Nos. 5,750,497 and 6,011,007, EP375437 and EP
`383472 and where such insulins may include, but are not
`limited to, NPH insulin, Lys B29 (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 (SemiLenteR) with
`extended insulin Zinc (UltralenteR), sold commercially as
`Lente R, insulin glargine (LantuSR) or extended insulin Zinc
`(UltralenteR), Lys' Pro’ human insulin (Huma-log(R),
`Asp human insulin, insulin aspart (Novolog(R), or a 30/70
`mixture of insulin aspart and insulin aspart protamine
`(NovoMix(R).
`0082 In one embodiment, the insulin is a derivative of
`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.
`0083. In one embodiment, the lysine residue to which the
`lipophilic substituent is attached is present at position B28 of
`the insulin peptide.
`0084. In an alternative embodiment, the lysine residue to
`which the lipophilic substituent is attached is present at
`position B29 of the insulin peptide.
`0085. In yet another embodiment, lipophilic substituent is
`an acyl group corresponding to a carboxylic acid having at
`least 6 carbon atoms.
`
`In another preferred embodiment, the lipophilic
`0086.
`Substituent is an acyl group, branched or unbranched, which
`corresponds to a carboxylic acid having a chain of carbon
`atoms 8 to 24 atoms long.
`0087. In another preferred embodiment, the lipophilic
`Substituent is an acyl group corresponding to a fatty acid
`having at least 6 carbon atoms.
`0088. In another preferred embodiment, the lipophilic
`Substituent is an acyl group corresponding to a linear,
`saturated carboxylic acid having from 6 to 24 carbon atoms.
`0089. In another preferred embodiment, the lipophilic
`Substituent is an acyl group corresponding to a linear,
`saturated carboxylic acid having from 8 to 12 carbon atoms.
`0090. In another preferred embodiment, the lipophilic
`Substituent is an acyl group corresponding to a linear,
`saturated carboxylic acid having from 10 to 16 carbon
`atOmS.
`0091. In another preferred embodiment, the lipophilic
`Substituent is an oligo oxyethylene group comprising up to
`10, preferably up to 5, oxyethylene units.
`0092. In another preferred embodiment, the lipophilic
`Substituent is an oligo oxypropylene group comprising up to
`10, preferably up to 5, oxypropylene units.
`0093. In one preferred embodiment, the invention relates
`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 amino acid 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' has a lipophilic substituent which
`comprises at least 6 carbon atoms; and 2-4 Zn" ions may be
`bound to each insulin hexamer with the proviso that when
`B30 is Thr or Ala and A21 and B3 are both ASn, and Phe'
`is not deleted