`
`US 20070010424Al
`
`c19) United States
`c12) Patent Application Publication
`Pedersen et al.
`
`c10) Pub. No.: US 2007/0010424 Al
`Jan. 11, 2007
`(43) Pub. Date:
`
`(54) PROPYLENE GLYCOL-CONTAINING
`PEPTIDE FORMULATIONS WHICH ARE
`OPTIMAL FOR PRODUCTION AND FOR
`USE IN INJECTION DEVICES
`
`(75)
`
`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) ................................ PA2003 01719
`
`Publication Classification
`
`(51)
`
`Int. Cl.
`A61K 38/28
`(2006.01)
`A61K 31/045
`(2006.01)
`A61K 38/26
`(2006.01)
`(52) U.S. Cl. ................................. 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(cid:173)
`tions comprising a peptide and propylene glycol, to methods
`of preparing such formulations, and to uses of such formu(cid:173)
`lations in the treatment of diseases and conditions for which
`use of the peptide contained in such formulations is indi(cid:173)
`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(cid:173)
`ment during production of a peptide formulation.
`
`MPI EXHIBIT 1016 PAGE 1
`
`MPI EXHIBIT 1016 PAGE 1
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0001
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 1 of 7
`
`US 2007/0010424 Al
`
`FIGURE 1
`
`MPI EXHIBIT 1016 PAGE 2
`
`MPI EXHIBIT 1016 PAGE 2
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0002
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 2 of 7
`
`US 2007/0010424 Al
`
`'"'.~
`.7: ,)I!
`;t,:'
`•.r ,:
`
`...,_.,..&WT.,.O\o
`
`Mannitol
`
`FIGURE 2
`
`r
`
`:,,:.O\ioe;" ••1
`....
`t,:~•'
`
`Argi-
`
`Ji
`
`·•::':
`...
`
`.,
`
`I
`'i't·l
`
`I if I
`
`lnasi-
`
`Glyce-
`
`MPI EXHIBIT 1016 PAGE 3
`
`MPI EXHIBIT 1016 PAGE 3
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0003
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 3 of 7
`
`US 2007/0010424 Al
`
`FIGURE 3
`
`Myo-inositol
`
`Maltose
`
`Glycerol
`
`MPI EXHIBIT 1016 PAGE 4
`
`MPI EXHIBIT 1016 PAGE 4
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0004
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 4 of 7
`
`US 2007/0010424 Al
`
`FIGURE4
`
`Glycine
`
`Lactose
`
`Mannitol
`
`MPI EXHIBIT 1016 PAGE 5
`
`MPI EXHIBIT 1016 PAGE 5
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0005
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 5 of 7
`
`US 2007/0010424 Al
`
`FIGURES
`
`MPI EXHIBIT 1016 PAGE 6
`
`MPI EXHIBIT 1016 PAGE 6
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0006
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 6 of 7
`
`US 2007/0010424 Al
`
`FIGURE 6
`
`MPI EXHIBIT 1016 PAGE 7
`
`MPI EXHIBIT 1016 PAGE 7
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0007
`
`
`
`Patent Application Publication Jan. 11, 2007 Sheet 7 of 7
`
`US 2007/0010424 Al
`
`FIGURE 7
`
`MPI EXHIBIT 1016 PAGE 8
`
`MPI EXHIBIT 1016 PAGE 8
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0008
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`1
`
`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 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(cid:173)
`ment during production of a peptide formulation.
`
`BACKGROUND OF THE INVENTION
`
`[0003] The inclusion of isotonicity agents in peptide(cid:173)
`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(cid:173)
`tive isotonic agent to mannitol for inclusion in peptide(cid:173)
`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(cid:173)
`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(cid:173)
`lated with any peptide and are also physically and chemi(cid:173)
`cally stable thus rendering them shelf-stable and suitable for
`invasive ( eg. injection, subcutaneous injection, intramuscu(cid:173)
`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(cid:173)
`maceutical formulation comprising a peptide and propylene
`glycol, where the propylene glycol is present in a concen(cid:173)
`tration of 1-100 mg/ml and the pH of the formulation is from
`
`7-10. In a preferred embodiment, the pharmaceutical for(cid:173)
`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(cid:173)
`servative, propylene glycol and buffer in water;
`
`[0010] b) preparing a second solution by dissolving the
`peptide in water;
`
`[0011] c) mixing the first and second solutions; and
`
`[0012] d) adjusting the pH of the mixture inc) 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(cid:173)
`servative and buffer in water;
`
`[0015] b) adding propylene glycol to the first solution;
`
`[0016] c) mixing the first solution with a second solu(cid:173)
`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(cid:173)
`ing a peptide formulation comprises:
`
`[0019] a) preparing a solution by dissolving preserva(cid:173)
`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(cid:173)
`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(cid:173)
`lene glycol-containing formulation relative to that observed
`for the formulation containing the previously utilized iso(cid:173)
`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(cid:173)
`ing the isotonicity agent previously utilized in said formu(cid:173)
`lation with propylene glycol at a concentration of between
`1-100 mg/ml.
`
`MPI EXHIBIT 1016 PAGE 9
`
`MPI EXHIBIT 1016 PAGE 9
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0009
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`2
`
`[0026]
`In one embodiment, the reduction in deposits in the
`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(cid:173)
`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 Arg34
`, Lys26(NE-(y(cid:173)
`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(cid:173)
`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 Arg34
`, Lys26(NE-(y-Glu(N"'-hexade(cid:173)
`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)(cid:173)
`containing Arg34
`, Lys26(W-(y-Glu(N"'-hexadecanoyl)))(cid:173)
`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(cid:173)
`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(cid:173)
`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(cid:173)
`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(cid:173)
`tides include, but are not limited to, glucagon, human growth
`hormone (hGH), insulin, aprotinin, FactorVII, tissue plas(cid:173)
`minogen activator (TPA), FactorVIIa, FFR-FactorVIIa,
`heparinase, ACTH, Heparin Binding Protein, corticotropin(cid:173)
`releasing factor, angio-tensin, calcitonin, glucagon-like pep(cid:173)
`tide-I, glucagon-like peptide-2, insulin-like growth factor-I,
`insulin-like growth factor-2, fibroblast growth factors, gas(cid:173)
`tric inhibitory peptide, growth hormone-releasing factor,
`pituitary adenylate cyclase activating peptide, secretin,
`enterogastrin, somatostatin, somatomedin, parathyroid hor(cid:173)
`mone, thrombopoietin, erythropoietin, hypothalamic releas(cid:173)
`ing factors, prolactin, thyroid stimulating hormones, endor(cid:173)
`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(cid:173)
`tors, growth factors and analogues as well as derivatives
`thereof where each of these peptides constitutes an alterna(cid:173)
`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(cid:173)
`dues have been added to the parent peptide. Such addition
`can take place either at the N-terminal end or at the C-ter(cid:173)
`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 (Kn) or a potency (EC50 ) of below 1 µM, 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
`
`MPI EXHIBIT 1016 PAGE 10
`
`MPI EXHIBIT 1016 PAGE 10
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0010
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`3
`
`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(cid:173)
`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(cid:173)
`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).
`
`[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(cid:173)
`logue, or a derivative of any of these.
`
`[0044]
`In one embodiment, the GLP-1 agonist is a deriva(cid:173)
`tive of GLP-1(7-36)-amide, GLP-1(7-37), a GLP-1(7-36)(cid:173)
`amide analogue or a GLP-1(7-37) analogue, which com(cid:173)
`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(cid:173)
`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(cid:173)
`prises a partially or completely hydrogenated cyclopen(cid:173)
`tanophenathrene skeleton.
`
`[0047]
`In another embodiment, the lipophilic substituent is
`a straight-chain or branched alkyl group.
`
`[0048]
`In yet another embodiment, the lipophilic substitu(cid:173)
`ent is an acyl group of a straight-chain or branched fatty
`acid. Preferably, the lipophilic substituent is an acyl group
`having the formula CH3 (CH2)nCO-, wherein n is an inte(cid:173)
`ger from 4 to 38, preferably an integer from 12 to 38, and
`most preferably is CH3 (CH2 ) 12CO-, CH3 (CH2 ) 14CO-,
`CH3 (CH2 ) 16CO-, CH3 (CH2 ) 18CO-, CH3 (CH2 ) 20CO(cid:173)
`and CH3 (CH2 ) 22CO-. 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(cid:173)
`chain or branched alkane a,w-dicarboxylic acid of the
`formula HOOC(CH2)mCO-, wherein mis an integer from
`4 to 38, preferably an integer from 12 to 38, and most
`preferably is HOOC(CH2 ) 14CO-, HOOC(CH2 ) 16CO-,
`or
`HOOC(CH2 ) 18CO-,
`HOOC(CH2 ) 20CO-
`HOOC(CH2 ) 22CO-.
`
`[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,
`
`[0052]
`(b) the carboxy group attached to the alpha(cid:173)
`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, Gin, 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(cid:173)
`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(cid:173)
`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, Gin, 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 ofLys 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
`
`MPI EXHIBIT 1016 PAGE 11
`
`MPI EXHIBIT 1016 PAGE 11
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0011
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`4
`
`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 NE -acylated lysine residue.
`
`[0064]
`is an
`spacer
`the
`In another embodiment,
`unbranched alkane a,w-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
`CH3 (CH2 )PNH----CO(CH2 )qCO-, 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
`CH3 (CH2 tCO-NHCH(COOH)(CH 2 ) 2CO-, 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
`CH3 (CH2 )sCO-NHCH((CH2 ) 2COOH)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(CH2 ),CO-wherein tis 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)(CH2 ) 4NH-CO(CH2 )uCH3 , 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
`CH3 (CH2 )vCO-NH-(CH2 ) 2 -CO, wherein vis 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)(CH2 ) 4NH(cid:173)
`COCH((CH2 ) 2COOH)NH----CO(CH2)wCH3 , 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)(CH2 ) 4NH(cid:173)
`CO(CH2 ) 2CH(COOH)NHCO(CH2 )xCH3 , 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
`Arg34
`, Lys 26(NE -( y-Glu(N"' -hexade-canoyl) ))-GLP-1 (7-37).
`
`[0074]
`In yet another embodiment the GLP-1 agonist is
`selected from the group consisting of Glys-GLP-1(7-36)(cid:173)
`amide, Glys -GLP-1 (7-37), Vals-GLP-1 (7-36)-amide, Vais -
`GLP-1 (7-37), Vais Asp22 -GLP-1 (7-36)-amide, Vais Asp 22
`-
`GLP-1(7-37), ValsGlu22-GLP-1(7-36)-amide, ValsGlu22
`-
`GLP-1(7-37), ValsLys22-GLP-1(7-36)-amide, ValsLys 22
`GLP-1(7-37), ValsArg22-GLP-1(7-36)-amide, ValsArg22
`GLP-1(7-37), ValsHis22-GLP-1(7-36)-amide, ValsHis 22
`-
`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 Arg26-GLP-1(7-37);
`34Lys 36
`Arg34-GLP-1(7-37); Lys 36-GLP-1(7-37); Arg26
`•
`-
`
`34Lys40 -GLP-
`34-GLP-1(7-37); Arg26
`GLP-1(7-37); Arg26
`'
`'
`1(7-37); Arg26Lys36-GLP-1(7-37); Arg34Lys36-GLP-1(7-
`MetsArg22-GLP-1(7-
`ValsArg22-GLP-1(7-37);
`37);
`3 7);GlysHis22-GLP-1(7-37);
`ValsHis 22-GLP-1(7-37);
`MetsHis 22-GLP-1(7-37); His37-GLP-1(7-37); Glys-GLP-
`1(7-37); Vals-GLP-1(7-37); Mets-GLP-1(7-37); GlysAsp 22
`-
`GLP-1(7-37); Val s Asp22 -GLP-1(7-37); Mets Asp22 -GLP-
`1(7-37); GlysG!u22-GLP-1(7-37); ValsGlu22-GLP-1(7-37);
`Met8Glu22-GLP-1(7-37);
`GlysLys 22-GLP-1(7-37);
`ValsLys22-GLP-1 (7-37);
`MetsLys 22-GLP-1(7-37);
`Glys Arg22-GLP-1 (7-37);
`ValsLys 22His 37 -GLP-1(7-37);
`GlysGlu22His 37 -GLP-1(7-37); ValsGlu22His37-GLP-1 (7-
`37); MetsGlu22His37 -GLP-1 (7-37); GlysLys22 His37 -GLP-
`1 (7-37); MetsLys22His37-GLP-1(7-37); Glys Arg22His 37
`-
`ValsArg22His 37 -GLP-1(7-37);
`GLP-1(7-37);
`Met8 Arg22His37-GLP-1 (7-37); GlysHis22His37-GLP-1 (7-
`37); ValsHis22His37 -GLP-1 (7-37); MetsHis 22His 37 -GLP-
`1 (7-37); GlysHis 37 -GLP-1(7-37); ValsHis 37 -GLP-1(7-37);
`Glys Asp 22His 37 -GLP-1(7-37);
`MetsHis 37 -GLP-1 (7-37);
`Vais Asp 22His 37 -GLP-1 (7-37); Mets Asp 22His37-GLP-1 (7-
`37); Arg26-GLP-1(7-36)-amide; Arg34-GLP-1(7-36)-amide;
`Lys 36-GLP-1(7-36)-amide;
`Arg26
`34Lys 36-GLP-1(7-36)(cid:173)
`'
`34-GLP-1 (7-36)-amide; Arg26
`amide; Arg26
`34Lys40 -GLP-
`•
`•
`Arg26Lys 36-GLP-1(7-36)-amide;
`1 (7-36)-amide;
`Arg34Lys 36 -GLP-1 (7-36)-amide; Glys-GLP-1 (7-36)-amide;
`Vals-GLP-1(7-36)-amide;
`Mets-GLP-1(7-36)-amide;
`Glys Asp 22-GLP-1(7-36)-amide; GlysG!u22His37-GLP-1 (7-
`36)-amide; Vais Asp22-GLP-1 (7-36)-amide; Mets Asp 22
`-
`GlysG!u22-GLP-1(7-36)-amide;
`GLP-1 (7-36)-amide;
`ValsGlu22-GLP-1(7-36)-amide; Met8Glu22-GLP-1(7-36)(cid:173)
`amide; GlysLys 22 -GLP-1 (7-36)-amide; Val sLys22 -GLP-1 (7-
`Met8Lys 22-GLP-1 (7-36)-amide;
`36)-amide;
`GlysHis 22His 37 -GLP-1 (7-36)-amide; Glys Arg22 -GLP-1 (7-
`36)-amide; Vais Arg22-GLP-1 (7-36)-amide; Mets Arg22
`-
`GlysHis 22-GLP-1 (7-36)-amide;
`GLP-1 (7-36)-amide;
`ValsHis22-GLP-1 (7-36)-amide; MetsHis 22-GLP-1(7-36)(cid:173)
`amide; His 37 -GLP-1 (7-36)-amide; Vais Arg22His 37 -GLP-
`1 (7-36)-amide; Mets Arg37 -GLP-1(7-36)-amide; GlysHis 37
`-
`ValsHis 37-GLP-1(7-36)-amide;
`GLP-1(7-36)-amide;
`MetsHis 37 -GLP-1 (7-36)-amide; Glys Asp 22 His37-GLP-1 (7-
`Vais Asp 22His 37-GLP-1(7-36)-amide;
`36)-amide;
`Mets Asp 22His37-GLP-1 (7-36)-amide;
`ValsGlu22His 37
`-
`GLP-1 (7-36)-amide; MetsG!u22His 37-GLP-1(7-36)-amide;
`GlysLys 22His 37 -GLP-1 (7-36)-amide; Val sLys22His 37 -GLP-
`MetsLys 22His 37-GLP-1(7-36)-amide;
`1 (7-36)-amide;
`Glys Arg22His37-GLP-1 (7-36)-amide; ValsHis22His 37 -GLP-
`1 (7-36)-amide; MetsHis 22His 37 -GLP-1 (7-36)-amide; and
`derivatives thereof.
`
`[0076]
`In yet another embodiment the GLP-1 agonist is
`selected from the group consisting ofValsTrp 19Glu22-GLP-
`1(7-37), ValsGlu22Val 25 -GLP-1(7-37), ValsTyr16Glu22
`-
`ValsTrp 16Glu22-GLP-1(7-37),
`GLP-1 (7-37),
`ValsLeu 16Glu22-GLP-1 (7-37), ValsTyr1sGlu22-GLP-1 (7-
`37), ValsGlu22 His37 -GLP-1(7-37), ValsGlu22Ile33 -GLP-
`Va!STrp 16Glu22Val25Ile33 -GLP-1(7-37),
`1 (7-37),
`Val sTrp 16Glu22Ile33 -GLP-1(7-37),
`ValsGlu22Val25Ile33
`-
`GLP-1 (7-37), ValsTrp 16Glu22Val 25 -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(cid:173)
`tives, and fragments thereof to be included within the
`
`MPI EXHIBIT 1016 PAGE 12
`
`MPI EXHIBIT 1016 PAGE 12
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1016-0012
`
`
`
`US 2007/0010424 Al
`
`Jan. 11, 2007
`
`5
`
`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
`and
`Y,
`HX1X2GTFITSDLSKQMEEEAVRLFIEWLKNGGPSSG
`APPPS; wherein X1X2=SD ( exendin-3) or GE ( exendin-4)).
`WO 97/46584 describes truncated versions of exendin pep(cid:173)
`tide(s). The disclosed peptides increase secretion and bio(cid:173)
`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(cid:173)
`logues stabilized by fusion to serum albumin or Fe portion
`of an lg are disclosed in WO 02/46227.
`
`[0079]
`In one embodiment, the exendin-4 analogue is
`HGEGTFTSDLSKQMEEEAVR(cid:173)
`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(cid:173)
`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(cid:173)
`tures thereof, where examples of insulin analogs and deriva(cid:173)
`tives are those disclosed in EP O 792 290 (Novo Nordisk
`A/S), EP O 214 826 and EP O 705 275 (Novo NordiskA/S),
`U.S. Pat. No. 5,504,188 (Eli Lilly), EP O 368 187 (Aventis),
`U.S. Pat. Nos. 5,750,497 and 6,011,007, EP 375437 and EP
`383472 and where such insulins may include, but are not
`(NE-tetradecanoyl)
`insulin, Lys ~29
`limited to, NPH
`des(B30) human insulin, LysB29-(NE -(y-glutamyl-N"'-litho(cid:173)
`cholyl) des(B30) human insulin, NLB29-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
`(Uitrnlente®), LysB 28 ProB29 human insulin (Huma-log®),
`Asp human insulin, insulin aspart (Novolog®), or a 30/70
`mixture of insulin aspart and insulin aspart protamine
`(NovoMix®).
`
`[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(cid:173)
`philic substituent is attached to the epsilon amino group of
`the lysine