(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(19) World Intellectual Property Organization
`International Bureau
`
`1111111111111111 IIIIII 111111111111111111111111111111111111111111111111111 IIII IIII IIII
`
`(43) International Publication Date
`9 January 2003 (09.01.2003)
`
`PCT
`
`(10) International Publication Number
`WO 03/002136 A2
`
`(51) International Patent Classification 7:
`
`A61K 38/00
`
`Richsvej, DK-2000 Frederiksberg (DK). ENGELUND,
`Dorthe, Kot; 39 Gassehaven, DK-2840 Holte (DK).
`
`(21) International Application Number: PCT/DK02/00437
`
`(22) International Filing Date:
`
`27 June 2002 (27.06.2002)
`
`(25) Filing Language:
`
`(26) Publication Language:
`
`English
`
`English
`
`(81) Designated States (national): AE, AG, AL, AM, AT, AU,
`AZ, BA, BB, BG, BR, BY, BZ, CA, CH, CN, CO, CR, CU,
`CZ, DE, DK, DM, DZ, EC, EE, ES, Fl, GB, GD, GE, GH,
`GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC,
`LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW,
`MX, MZ, NO, NZ, OM, PH, PL, PT, RO, RU, SD, SE, SG,
`SI, SK, SL, TJ, TM, TN, TR, TT, TZ, UA, UG, UZ, VN,
`YU, ZA, ZM, ZW.
`
`(30) Priority Data:
`PA2001 01010
`PA 2001 01011
`PA 2001 01053
`PA 2001 01052
`PA 2002 00093
`PA 2002 00092
`
`28 June 2001 (28.06.2001)
`28 June 2001 (28.06.2001)
`4 July 2001 (04.07.2001)
`4 July 2001 (04.07.2001)
`18 January 2002 (18.01.2002)
`18 January 2002 (18.01.2002)
`
`DK
`DK
`DK
`DK
`DK
`DK
`
`(84) Designated States (regional): ARIPO patent (GH, GM,
`KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZM, ZW),
`Eurasian patent (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM),
`European patent (AT, BE, CH, CY, DE, DK, ES, Fl, FR,
`GB, GR, IE, IT, LU, MC, NL, PT, SE, TR), OAPI patent
`(BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, ML, MR,
`NE, SN, TD, TG).
`
`(71) Applicant: NOVO NORDISKA/S [DK/DK]; Novo Alie,
`DK-2880 Bagsvaerd (DK).
`
`Published:
`without international search report and to be republished
`upon receipt of that report
`
`(72) Inventors: FLINK, James, M; 76, st.th. Taarbaek
`Strandvej, DK-2930 Klampenborg (DK). LARSEN, Silke,
`Moller; 10, 1.mf, Ejgaardsvej, DK-2920 Charlottenlund
`(DK). JENSEN, Simon, Bjerregaard; 103A, 5.th., C.F.
`
`For two-letter codes and other abbreviations, refer to the "Guid(cid:173)
`ance Notes on Codes and Abbreviations" appearing at the begin(cid:173)
`ning of each regular issue of the PCT Gazette.
`
`iiiiiiii
`
`-iiiiiiii
`
`iiiiiiii --------
`---iiiiiiii
`iiiiiiii ----
`
`\0
`~
`,-...I
`M
`0
`
`0 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
`
`0 ---~
`o (54) Title: STABLE FORMULATION OF MODIFIED GLP-1
`~ (57) Abstract: Pharmaceutical formulations of GLP-1 compounds and methods for preparation thereof.
`
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`WO 03/002136
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`PCT/DK02/00437
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`STABLE FORMULATION OF MODIFIED GLP-1
`
`1
`
`Field of the invention
`
`The present invention relates to pharmaceutical formulations comprising GLP-1
`
`5
`
`compounds, uses thereof and methods for preparing said formulations.
`
`Background of the invention
`
`Peptides are widely used in medical practice, and since they can be produced by re(cid:173)
`
`combinant DNA technology it can be expected that their importance will increase also in the
`
`10
`
`years to come.
`
`15
`
`20
`
`25
`
`30
`
`The hormones regulating insulin secretion belong to the so-called enteroinsular
`
`axis, designating a group of hormones, released from the gastrointestinal mucosa in re(cid:173)
`sponse to the presence and absorption of nutrients in the gut, which promote an early and
`potentiated release of insulin. The enhancing effect on insulin secretion, the so-called incretin
`effect, is probably essential for a normal glucose tolerance. Many of the gastrointestinal hor(cid:173)
`mones, including gastrin and secretin (cholecystokinin is not insulinotropic in man), are insu(cid:173)
`linotropic, but the only physiologically important ones, those that are responsible for the in(cid:173)
`cretin effect, are the glucose-dependent insulinotropic polypeptide, GIP, and glucagon-like
`peptide-1 (GLP-1 ). Because of its insulinotropic effect, GIP, isolated in 1973 immediately at-
`tracted considerable interest among diabetologists. However, numerous investigations car(cid:173)
`ried out during the following years clearly indicated that a defective secretion of GIP was not
`involved in the pathogenesis of insulin dependent diabetes mellitus (IDDM) or non insulin(cid:173)
`dependent diabetes mellitus (NIDDM). Furthermore, as an insulinotropic hormone, GIP was
`found to be almost ineffective in NIDDM. The other incretin hormone, GLP-1 is the most po-
`tent insulinotropic substance known. Unlike GIP, it is surprisingly effective in stimulating insu(cid:173)
`lin secretion in NIDDM patients. In addition, and in contrast to the other insulinotropic hor(cid:173)
`
`mones (perhaps with the exception of secretin) it also potently inhibits glucagon secretion.
`Because of these actions it has pronounced blood glucose lowering effects particularly in pa(cid:173)
`tients with NIDDM.
`GLP-1, a product of the proglucagon, is one of the youngest members of the se-
`
`cretin-VIP family of peptides, but is already established as an important gut hormone with
`'
`regulatory function in glucose metabolism and gastrointestinal secretion and metabolism.
`
`'
`
`The glucagon gene is processed differently in the pancreas and in the intestine. In the pan(cid:173)
`
`creas, the processing leads to the formation and parallel secretion of 1) glucagon itself, oc-
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`PCT/DK02/00437
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`2
`
`cupying positions 33-61 of proglucagon (PG); 2) an N-terminal peptide of 30 amino acids
`
`(PG (1-30)) often called glicentin-related pancreatic peptide, GRPP; 3) a hexapeptide corre(cid:173)
`
`sponding to PG (64-69); 4) and, finally, the so-called major proglucagon fragment (PG (72-
`
`158)), in which the two glucagon-like sequences are buried. Glucagon seems to be the only
`
`5
`
`biologically active product. in contrast, in the intestinal mucosa, it is glucagon that is buried in
`
`a larger molecule, while the two glucagon-like peptides are formed separately.
`
`While much attention has been focused on the pharmacological properties of acy(cid:173)
`
`lated GLP-1 compounds, hitherto little is known about their physico-chemical and solution
`
`structural properties. Such knowledge is a prerequisite for rational handling during e.g. pro-
`
`1 o
`
`duction, purification and formulation work and is eventually important for understanding of the
`
`structural basis for the protraction mechanism.
`It is an important technical challenge to ensure prolonged stability during storage
`
`(shelf life) of many protein based drug products due to the inherent !ability of macromole(cid:173)
`
`cules. Hence, proteins are sensitive to both chemical and physical degradation unlike many
`
`15
`
`small molecules. Chemical degradation involves covalent bonds, such as hydrolysis, racemi(cid:173)
`
`zation, oxidation or crosslinking. Physical degradation involves conformational changes rela(cid:173)
`
`tive to the native structure, which includes loss of higher order structure, aggregation, precipi(cid:173)
`
`tation or adsorption to surfaces. GLP-1 is known to be prone to instability due to aggregation.
`
`Both degradation pathways may ultimately lead to loss of biological activity of the protein
`
`20
`
`drug.
`
`GLP-1 and analogues of GLP-1 and fragments thereof are potentially useful i.a. in the
`
`treatment of type 1 and type 2 diabetes. However, solubility limitations and the low stability
`
`against the actions of endogenous diaminopeptidyl peptidase limits the usefulness of these
`
`compounds, and thus there still is a need for improvements in this field.
`
`25
`
`In WO 99/43341 are disclosed certain pharmaceutical formulations comprising GLP-1
`
`having a lipophilic substituent. All of the disclosed formulations are maintained at pH 7.4.
`
`In WO 00/37098 are disclosed shelf-stable formulations comprising GLP-1, a preserva(cid:173)
`
`tive, and a tonicity modifier, at pH 8.2 to 8.8.
`
`30
`
`Human GLP-1 is a 37 amino acid residue peptide originating from preproglucagon which
`
`is synthesised i.a. in the L-cells in the distal ileum, in the pancreas and in the brain. Processing
`
`of preproglucagon to give GLP-1(7-36)amide, GLP-1(7-37) and GLP-2 occurs mainly in the L(cid:173)
`
`cells. A simple system is used to describe fragments and analogues of this peptide. Thus, for
`example, Val 8-GLP-1 (7-37) (or Val8GLP-1 (7-37)) designates a fragment of GLP-1 formally de-
`
`35
`
`rived from GLP-1 by deleting the amino acid residues Nos. 1 to 6 and substituting the naturally
`
`occurring amino acid residue in position 8 (Ala) by Val. Similarly, Lys34(N 8-tetradecanoyl)-GLP-
`
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`3
`
`1 (7-37) designates GLP-1 (7-37) wherein the a-amino group of the Lys residue in position 34 has
`
`been tetradecanoylated. For convenience the amino acid sequence of GLP-1 (7-37) is given
`
`below, wherein the N-terminal His is no. 7 and the C-terminal Gly is no. 37:
`
`5
`
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser(cid:173)
`
`Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-
`
`1 le-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly.
`
`Where reference in this text is made to C-terminally extended GLP-1 analogues, the amino acid
`
`residue in position 38 is Arg unless otherwise indicated, the optional amino acid residue in
`
`position 39 is also Arg unless otherwise indicated and the optional amino acid residue in
`
`position 40 is Asp unless otherwise indicated. Also, if a C-terminally extended analogue extends
`
`to position 41, 42, 43, 44 or 45, the amino acid sequence of this extension is as in the
`
`corresponding sequence in human preproglucagon unless otherwise indicated.
`
`1 o
`
`15
`
`Summary of the invention
`
`We have discovered that certain modified GLP-1 or analogues thereof when formu(cid:173)
`
`lated in aqueous solution together with a buffer, are physically stable at high concentrations
`
`of the modified GLP-1 or analogues thereof, when kept in the pH range from about 7 to about
`
`20
`
`10. The present formulations are physically stable within a given shelf life period at the rec(cid:173)
`
`ommended storage temperature (typically 2-3 years at 2-8°C). Furthermore, the present for(cid:173)
`
`mulations are physically stable during in-use (typically 1 month at accelerated temperatures
`
`e.g. 25°C or 37°C). The formulations of the invention are also chemically stable thus render(cid:173)
`
`ing them shelf-stable and suitable for invasive (eg. injection, subcutaneous injection, intra-
`
`25 muscular, intraveneous or infusion) as well as non-invasive (eg nasal or pulmonary, trans(cid:173)
`
`dermal or transmucosal e.g. buccal) means of administration. When the inventive formula(cid:173)
`
`tion comprising a GLP-1 compound was compared to the same formulation comprising GLP-
`
`1 (7-37) substituted for the GLP-1 compound, the physical stability was increased considera(cid:173)
`
`bly, and typically the shelf-life was increased from a few seconds to several months in the
`
`30
`
`tests used.
`
`One object of the present invention is to provide a pharmaceutical formulation com(cid:173)
`
`prising a GLP-1 compound, and a buffer, wherein said GLP-1 compound is GLP-1(7-37) or
`
`an analogue thereof wherein an amino acid residue of the parent peptide has a lipophilic sub-
`
`35
`
`stituent attached optionally via a spacer, wherein said GLP-1 compound is present in a con-
`
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`PCT/DK02/00437
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`4
`
`centration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to
`
`1 O;
`
`Another object of the present invention is to provide a method of preparing a physi(cid:173)
`
`cally stable pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 com-
`
`5
`
`pound is GLP-1(7-37) or an analogue thereof wherein an amino acid residue of the parent
`
`peptide has a lipophilic substituent attached optionally via a spacer, comprising preparing a
`
`formulation containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`present in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a
`
`1 o
`
`15
`
`pH from 7.0 to 10.
`
`In one aspect of the invention the formulation contains a GLP-1 compound in a con-
`
`centration from 1 mg/ml to 100 mg/ml.
`
`In another aspect of the invention the formulation has a pH from 7 .5 to 10.
`, Lys26(N-s-(y-Glu(N-a(cid:173)
`In one embodiment the GLP-1 compound is Arg 34
`
`hexadecanoyl)))-GLP-1 (7-37).
`
`Description of the invention
`
`In one aspect the invention relates to a pharmaceutical formulation comprising a GLP-1
`
`compound, and a buffer, wherein said GLP-1 compound is GLP-1 (7-37) or an analogue
`
`thereof wherein an amino acid residue of the parent peptide has a lipophilic substituent attached
`
`20
`
`optionally via a spacer, wherein said GLP-1 compound is present in a concentration from 0.1
`
`mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to 10;
`
`provided that if an isotonic agent is present and pH is 7.4 then mannitol or NaCl is not the
`
`isotonic agent.
`
`In another aspect the invention relates to a pharmaceutical formulation comprising a
`
`25 GLP-1 compound, and a buffer, wherein said GLP-1 compound is GLP-1 (7-37) or an ana(cid:173)
`
`logue thereof wherein an amino acid residue of the parent peptide has a lipophilic substituent
`
`attached optionally via a spacer, wherein said GLP-1 compound is present in a concentration
`
`from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to 10;
`
`provided that if an isotonic agent is present and pH is 7.4 then mannitol or NaCl is not the
`
`30
`
`isotonic agent.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising a
`
`GLP-1 compound, and a buffer, wherein said GLP-1 compound is GLP-1 (7-37) or an ana(cid:173)
`
`logue thereof wherein an amino acid residue of the parent peptide has a lipophilic substituent
`
`attached optionally via a spacer, wherein said GLP-1 compound is present in a concentration
`
`35
`
`from 0.1 mg/ml or above, and wherein said formulation has a pH from 7.0 to 10.
`
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`WO 03/002136
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`PCT/DK02/00437
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`5
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising a
`
`GLP-1 compound, and a buffer, wherein said GLP-1 compound is GLP-1(7-37) or an ana(cid:173)
`
`logue thereof wherein an amino acid residue of the parent peptide has a lipophilic substituent
`
`attached optionally via a spacer, wherein said GLP-1 compound is present in a concentration
`
`5
`
`from 1 mg/ml or above, and wherein said formulation has a pH from 7.0 to 10.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`a GLP-1 compound, and a buffer, wherein said GLP-1 compound is GLP-1 (7-37) or an ana(cid:173)
`
`logue thereof, wherein an amino acid residue of the parent peptide has a lipophilic substituent
`
`attached optionally via a spacer, wherein said GLP-1 compound is present in a concentration
`
`1 o
`
`from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to 10.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`a GLP-1 compound, and a buffer, wherein said GLP-1 compound is GLP-1 (7-37) or an ana(cid:173)
`
`logue thereof, wherein an amino acid residue of the parent peptide has a lipophilic substituent
`
`attached optionally via a spacer, wherein said GLP-1 compound is present in a concentration
`
`15
`
`from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising prepararing a formulation
`
`20
`
`containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present in a
`
`concentration from 0.1 mg/ml or above, and wherein said formulation has a pH from 7.0 to
`
`10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`25
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising prepararing a formulation
`
`containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present in a
`
`concentration from 1 mg/ml or above, and wherein said formulation has a pH from 7.0 to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`30
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`lipophilic substituent attached optionally via a spacer, comprising prepararing a formulation
`
`containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present in a
`
`concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0
`
`35
`
`to 10.
`
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`WO 03/002136
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`PCT/DK02/00437
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`6
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con-
`
`5
`
`taining the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present in a
`concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to
`
`10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`10
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con(cid:173)
`
`taining the GLP-1 compound, water, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.0 to 10.
`
`15
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con(cid:173)
`
`taining the GLP-1 compound, water, and a buffer, wherein said GLP-1 compound is present
`
`20
`
`in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.0 to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`25
`
`lipophilic substituent attached optionally via a spacer, comprising preparing an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.0 to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`30
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`35
`
`7.0 to 10.
`
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`PCT/DK02/00437
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`7
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acicl residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con-
`
`5
`
`taining the GLP-1 compound, water, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.5to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 o
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con(cid:173)
`
`taining the GLP-1 compound, water, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.5 to 10.
`
`15
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`20
`
`in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.5to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`25
`
`lipophilic substituent attached optionally via a spacer, comprising preparing an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.5 to 10.
`
`30
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con(cid:173)
`
`taining the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present in a
`
`35
`
`concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0
`
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`

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`WO 03/002136
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`PCT/DK02/00437
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`8
`
`to 1 0; provided that if an isotonic agent is present and pH is 7.4 then mannitol or NaCl is not
`
`the isotonic agent.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`5
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, comprising preparing a formulation con(cid:173)
`
`taining the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present in a
`
`concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from 7.0 to
`
`1 0; provided that if an isotonic agent is present and pH is 7.4 then mannitol or NaCl is not the
`
`1 o
`
`isotonic agent.
`
`In one embodiment of the invention the pharmaceutical formulation is an aqueous
`
`formulation, i.e. a formulation comprising water. Such formulation is typically a solution or a
`
`suspension. In a further embodiment of the invention the pharmaceutical formulation is an
`
`aqueous solution. The term "aqueous formulation" is defined as a formulation comprising at
`
`15
`
`least 50 %w/w water. Likewise, the term "aqueous solution" is defined as a solution compris(cid:173)
`
`ing at least 50 %w/w water, and the term "aqueous suspension" is defined as a suspension
`
`comprising at least 50 %w/w water.
`
`In another embodiment the pharmaceutical formulation is a freeze-dried formulation,
`
`whereto the physician or the patient adds the solvent prior to use.
`
`20
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`GLP-1(7-37) or an analogue thereof wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, wherein said GLP-1 compound is pre(cid:173)
`
`sent in a concentration from 0.1 mg/ml or above, and wherein said formulation has a pH from
`
`25
`
`7.0 to 10.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`GLP-1(7-37) or an analogue thereof wherein an amino acid residue of the parent peptide has a
`
`lipophilic substituent attached optionally via a spacer, wherein said GLP-1 compound is pre-
`
`30
`
`sent in a concentration from 1 mg/ml or above, and wherein said formulation has a pH from
`
`7.0to 10.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`GLP-1(7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has
`
`35
`
`a lipophilic substituent attached optionally via a spacer, wherein said GLP-1 compound is pre-
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1004, p. 9 of 50
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`WO 03/002136
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`PCT/DK02/00437
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`9
`
`sent in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH
`
`from 7.0 to 10.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`5 GLP-1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has
`
`a lipophilic substituent attached optionally via a spacer, wherein said GLP-1 compound is pre(cid:173)
`
`sent in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH
`
`from 7.0 to 10.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`1 o
`
`an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`GLP-1(7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has
`
`a lipophilic substituent attached optionally via a spacer, wherein said GLP-1 compound is pre(cid:173)
`
`sent in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH
`from 7. 0 to 1 O; provided that if an isotonic agent is present and pH is 7.4 then mannitol or
`
`15
`
`NaCl is not the isotonic agent.
`
`In a further aspect the invention relates to a pharmaceutical formulation comprising
`
`an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`GLP-1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has
`
`a lipophilic substituent attached optionally via a spacer, wherein said GLP-1 compound is pre-
`
`20
`
`sent in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH
`
`from 7. 0 to 1 O; provided that if an isotonic agent is present and pH is 7.4 then mannitol or
`
`NaCl is not the isotonic agent.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`25
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a lipo(cid:173)
`
`philic substituent attached optionally via a spacer, comprising preparation of an aqueous
`
`solution containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`present in a concentration from 0.1 mg/ml or above, and wherein said formulation has a pH
`
`from 7.0 to 10.
`
`30
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a lipo(cid:173)
`
`philic substituent attached optionally via a spacer, comprising preparation of an aqueous
`
`solution containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is
`
`35
`
`present in a concentration from 1 mg/ml or above, and wherein said formulation has a pH
`
`from 7.0 to 10.
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1004, p. 10 of 50
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`

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`WO 03/002136
`
`PCT/DK02/00437
`
`10
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a lipo(cid:173)
`
`philic substituent attached optionally via a spacer, comprising preparation of an aqueous solu-
`
`5
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.0to 10.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 o
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a lipo(cid:173)
`
`philic substituent attached optionally via a spacer, comprising preparation of an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.0 to 10.
`
`15
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a lipo(cid:173)
`
`philic substituent attached optionally via a spacer, comprising preparation of an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`20
`
`in a concentration from 0.1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7.0 to 10; provided that if an isotonic agent is present and_pH is 7.4 then mannitol or NaCl is
`
`not the isotonic agent.
`
`In a further aspect the invention relates to a method of preparing a physically stable
`
`pharmaceutical formulation of a GLP-1 compound wherein said GLP-1 compound is GLP-
`
`25
`
`1 (7-37) or an analogue thereof, wherein an amino acid residue of the parent peptide has a lipo(cid:173)
`
`philic substituent attached optionally via a spacer, comprising preparation of an aqueous solu(cid:173)
`
`tion containing the GLP-1 compound, and a buffer, wherein said GLP-1 compound is present
`
`in a concentration from 1 mg/ml to 100 mg/ml, and wherein said formulation has a pH from
`
`7. 0 to 1 0; provided that if an isotonic agent is present and pH is 7.4 then mannitol or NaCl is
`
`30
`
`not the isotonic agent.
`
`In a further aspect the present invention relates to a method of reducing blood glu(cid:173)
`
`cose levels, treating diabetes type I, diabetes type II, obesity, or inhibiting gastric acid secre(cid:173)
`
`tion, inhibiting apoptosis of 13-cells, or stimulating the proliferation of 13-cells, comprising ad(cid:173)
`
`ministering to a patient in need thereof an effective amount of a pharmaceutical formulation
`
`35
`
`comprising an aqueous solution of a GLP-1 compound, and a buffer, wherein said GLP-1
`
`compound is GLP-1(7-37) or an analogue thereof wherein an amino acid residue of the parent
`
` PFIZER, INC. v. NOVO NORDISK A/S - IP