`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`International Bureau
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`WO 00/37098
`(51) International Patent Classification 7 :
`A61K 38/26, 38/28, 47/10, 47/26, A61P
`3/10 // (A61K 38/28, 38:26)
`
`(43) International Publication Date:
`
`29 June 2000 (29.06.00)
`
`(11) International Publication Number:
`
`Al
`
`(21) International Application Number:
`
`PCT/US99/30395
`
`(22) International Filing Date:
`
`21 December 1999 (21.12.99)
`
`(30) Priority Data:
`60/113,499
`
`22 December 1998 (22.12.98)
`
`US
`
`(71) Applicant (for all designated States except US): ELI LILLY
`AND COMPANY [US/US]; Lilly Corporate Center, Indi(cid:173)
`anapolis, IN 46285 (US).
`
`(72) Inventors; and
`(75) Inventors/Applicants (for US only): BRADER, Mark, L.
`[NZ/US]; 6465 North Park Avenue, Indianapolis, IN 46220
`(US). PEKAR, Allen, H. [US/US]; 5354 North Park Avenue,
`Indianapolis, IN 46220 (US).
`
`(74) Agent: MACIAK, Ronald, S.; Eli Lilly and Company, Lilly
`Corporate Center, Drop Code 1501, Indianapolis, IN 46285
`(US).
`
`(81) Designated States: AE, AL, AM, AT, AU, AZ, BA, BB, BG,
`BR, BY, CA, CH, CN, CR, CU, CZ, DE, DK, DM, EE, ES,
`Fl, GB, GD, GE, 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, NO, NZ, PL, PT, RO, RU, SD, SE,
`SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ,
`VN, YU, ZA, ZW, ARIPO patent (GH, GM, KE, LS, MW,
`SD, SL, SZ, TZ, UG, 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), OAPI patent (BF, BJ, CF, CG, CI, CM, GA, GN,
`GW, ML, MR, NE, SN, TD, TG).
`
`Published
`With international search report.
`Before the expiration of the time limit for amending the
`claims and to be republished in the event of the receipt of
`amendments.
`
`(54) Title: SHELF-STABLE FORMULATION OF GLUCAGON-LIKE PEPTIDE-I
`
`(57) Abstract
`
`Glucagon-like peptide-I (GLP-1) has been shown to be useful in the treatment of diabetes. The invention encompasses a shelf
`stable formulation that comprises a therapeutically effective amount of GLP-1, a pharmaceutically acceptable preservative, and a tonicity
`modifier, and that has a pH between about 8.2 to about 8.8.
`
`MPI EXHIBIT 1074 PAGE 1
`
`MPI EXHIBIT 1074 PAGE 1
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0001
`
`
`
`FOR THE PURPOSES OF INFORMATION ONLY
`
`Codes used to identify States party to the PCT on the front pages of pamphlets publishing international applications under the PCT.
`
`AL
`AM
`AT
`AU
`AZ
`BA
`BB
`BE
`BF
`BG
`BJ
`BR
`BY
`CA
`CF
`CG
`CH
`CI
`CM
`CN
`cu
`CZ
`DE
`DK
`EE
`
`Albania
`Annenia
`Austria
`Australia
`Azerbaijan
`Bosnia and Herzegovina
`Barbados
`Belgium
`Burkina Faso
`Bulgaria
`Benin
`Brazil
`Belarus
`Canada
`Central African Republic
`Congo
`Switzerland
`C6te d'Ivoire
`Cameroon
`China
`Cuba
`Czech Republic
`Gennany
`Denmark
`Estonia
`
`ES
`Fl
`FR
`GA
`GB
`GE
`GH
`GN
`GR
`HU
`IE
`IL
`IS
`IT
`JP
`KE
`KG
`KP
`
`KR
`KZ
`LC
`LI
`LK
`LR
`
`Spain
`Finland
`France
`Gabon
`United Kingdom
`Georgia
`Ghana
`Guinea
`Greece
`Hungary
`Ireland
`Israel
`Iceland
`Italy
`Japan
`Kenya
`Kyrgyzstan
`Democratic People's
`Republic of Korea
`Republic of Korea
`Kazakstan
`Saint Lucia
`Liechtenstein
`Sri Lanka
`Liberia
`
`LS
`LT
`LU
`LV
`MC
`MD
`MG
`MK
`
`ML
`MN
`MR
`MW
`MX
`NE
`NL
`NO
`NZ
`PL
`PT
`RO
`RU
`SD
`SE
`SG
`
`Lesotho
`Lithuania
`Luxembourg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`The fonner Yugoslav
`Republic of Macedonia
`Mali
`Mongolia
`Mauritania
`Malawi
`Mexico
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Singapore
`
`SI
`SK
`SN
`sz
`TD
`TG
`TJ
`TM
`TR
`TT
`UA
`UG
`us
`uz
`VN
`YU
`zw
`
`Slovenia
`Slovakia
`Senegal
`Swaziland
`Chad
`Togo
`Tajikistan
`Turkmenistan
`Turkey
`Trinidad and Tobago
`Ukraine
`Uganda
`United States of America
`Uzbekistan
`Viet Nam
`Yugoslavia
`Zimbabwe
`
`MPI EXHIBIT 1074 PAGE 2
`
`MPI EXHIBIT 1074 PAGE 2
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0002
`
`
`
`WO 00/37098
`
`PCT/US99/30395
`
`SHELF-STABLE FORMULATION OF GLUCAGON-LIKE PEPTIDE-I
`
`Background of the Invention
`Glucagon-like peptide-I (7-37)-OH (GLP-1) is a 31 amino acid hormone that is
`produced by post-translational processing of the preglucagon gene product in the brain,
`stomach, intestine, and pancreas. The main physiological function of GLP-1 is to regulate
`insulin secretion in response to glucose, and thus it has the ability to normalize blood
`glucose levels. As such, there has been interest in GLP-1, its analogs and derivatives as
`potential therapeutic agents for the treatment of diabetes. A particular advantage to the use
`of GLP-1 over other drugs in the treatment of diabetes is that administration of GLP-1 at
`doses in the 1-5 nmole range exhibit few adverse side effects, such as hypoglycemia.
`Unexpectedly, GLP-1 also has been shown to work in patients that have secondary failure
`to sulfonylurea drugs, the most common drug type for the treatment of type II diabetes.
`GLP-1 also is a potent inhibitor of gastric acid secretion and gastric emptying.
`In general, effective therapeutic administration of peptides can be problematic since
`peptides often are degraded in the gastrointestinal tract by various peptidases. Additionally,
`certain peptide treatment protocols require either continuous or repeated administration of
`the peptide agent over an extended period of time. Repeated injections cause both
`inconvenience and discomfort to the user. Thus, chronic use of the peptide agent, which
`would be required for patients afflicted with diabetes, would result in inconvenience and
`discomfort to the user.
`The long-term stability of peptides, particularly GLP-1, as components of a
`pharmaceutical composition for administration to mammals, is questionable. Such a lack of
`stability adversely affects bioavailability. In fact, when stored at low temperatures of 4° C,
`by-products of GLP-1(7-37) have been found as early as eleven months after sample
`preparation (see Mojsov, Int. J. Peptide Protein Res., Vol. 40, pages 333-343 (1992)).
`Additionally, the biological half-life of GLP-1 molecules, particularly those molecules
`affected by the activity of dipeptidyl-peptidase IV (DPPIV), is quite short. For example,
`
`MPI EXHIBIT 1074 PAGE 3
`
`MPI EXHIBIT 1074 PAGE 3
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0003
`
`
`
`WO 00/37098
`PCT /US99/30395
`the biological half-life of GLP-1(7-37) is only 3 to 5 minutes (see U.S. Patent No.
`5,118,666), which is further augmented by its rapid absorption following parenteral
`administration to a mammal.
`Another factor decreasing the bioavailability of GLP-1 is the solubility of GLP-1
`when incorporated into an aqueous solution. The solubility of GLP-1 is highly dependent
`on the environment, such as the choice of buffering system, and the treatment that the
`peptide has undergone. For example, conversion of a peptide into its salt form plays a role
`in its solubility. In this regard, synthetic GLP-1 is highly soluble in neutral phosphate
`buffered saline. Because the solubility of the peptide is high in such aqueous solutions,
`slow release of the peptide can be difficult to attain unless the peptide is incorporated into a
`system for slow release.
`Stable formulations of therapeutic agents are particularly required for use in delivery
`devices that expose these agents to elevated temperatures and/or mechanical stress. For
`example, stable GLP-1 formulations are required for use in continuous infusion systems
`and pen delivery devices. Current formulations provide only limited stability in these types
`of delivery devices.
`In continuous infusion systems, a fluid containing a therapeutic agent is pumped
`from a reservoir, usually to a subcutaneous, intravenous, or intraperitoneal depot. The
`reservoir, which must be refilled periodically, is attached to the patient's body, or is
`implanted in the patient's body. In either case, the patient's body heat and body motion,
`and turbulence in the tubing and pump impart a relatively high amount of thermo(cid:173)
`mechanical energy to the formulation. In the interest of minimizing the frequency with
`which the reservoir is refilled, and of minimizing the size of the reservoir, formulations
`having a relatively high concentration of the therapeutic agent are advantageous.
`Injector pens also have been developed to allow diabetic patients to accurately
`measure and administer controlled doses of insulinotropic agents. Generally, these pens are
`secured onto a cartridge having a particular quantity of liquid medication sealed therein.
`The cartridge includes a plunger and a mechanism for advancing the plunger in the
`cartridge in such a manner to dispense the medication. Injector pens may be reusable or
`disposable. In reusable pens, a user can change a spent cartridge and reset the leadscrew of
`the pen back to its initial position. In a disposable pen, the cartridge is permanently
`
`-2-
`
`MPI EXHIBIT 1074 PAGE 4
`
`MPI EXHIBIT 1074 PAGE 4
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0004
`
`
`
`WO 00/37098
`
`PCT /US99/30395
`
`embodiment, the GLP-1 molecule of the formulation is a derivative of GLP-1 and is
`selected from the group consisting of a peptide having the amino acid sequence:
`NH2-His7-Ala-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Glu-Gly-Gln-Ala-
`Ala25-Lys-Glu-Phe-Ile-Ala30-Trp-Leu-Val-X
`(SEQ ID N0:3)
`and a pharmaceutically-acceptable salt thereof, wherein X is selected from the group
`consisting of Lys and Lys-Gly; a pharmaceutically-acceptable lower alkylester of the peptide;
`and a pharmaceutically-acceptable amide of the peptide selected from the group consisting of
`amide, lower alkyl amide, and lower dialkyl amide. In another preferred embodiment, the
`formulation also comprises a long-acting insulin agent.
`The present invention also provides a method of enhancing the expression of insulin
`in a mammalian pancreatic ~-type islet cell in need of such enhancement, comprising
`administering to the cell, an effective amount of a shelf-stable pharmaceutical formulation,
`wherein the formulation comprises a therapeutically effective amount of a GLP-1 molecule,
`a pharmaceutically acceptable preservative, and a tonicity modifier, and wherein the
`formulation has a pH that is about 8.2 to about 8.8. In a preferred embodiment, the
`formulation used in the therapeutic method comprises a buffer, such as TRIS. In another
`preferred embodiment, the formulation used in the therapeutic method further comprises a
`surfactant, such as Brij-35. In an additional preferred embodiment, the GLP-1 molecule of
`the formulation thus administered is an analog of GLP-1 and is selected from the group
`consisting of a peptide having the amino acid sequence:
`R1-X-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Y-Gly-Gln-Ala-Ala25
`Lys-Z-Phe-Ile-Ala30-Trp-Leu-Val-Lys-Gly35-Arg-R2
`(SEQ ID N0:2)
`and a pharmaceutically-acceptable salt thereof, wherein R1 is His or desamino-histidine, X
`is Ala, Gly or Val, Y is Glu or Gin, Z is Glu or Gin and R2 is Gly-OH. In an especially
`preferred embodiment, the GLP-1 molecule administered is according to SEQ ID NO: 2,
`wherein R1 is L-histidine, Xis Val, Y is Glu, Z is Glu, and R2 is Gly-OH. In an alternative
`preferred embodiment, the GLP-1 molecule administered is a derivative of GLP-1 and is
`selected from the group consisting of a peptide having the amino acid sequence:
`NH2-His7-Ala-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Glu-Gly-Gln-
`Ala-Ala25-Lys-Glu-Phe-Ile-Ala30-Trp-Leu-Val-X
`(SEQ ID N0:3)
`and a pharmaceutically-acceptable salt thereof, wherein X is selected from the group
`consisting of Lys and Lys-Gly; a pharmaceutically-acceptable lower alkylester of the peptide;
`
`-
`
`-4-
`
`MPI EXHIBIT 1074 PAGE 5
`
`MPI EXHIBIT 1074 PAGE 5
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0005
`
`
`
`WO 00/37098
`
`PCT /US99/30395
`
`and a pharmaceutically-acceptable amide of the peptide selected from the group consisting of
`amide, lower alkyl amide, and lower dialkyl amide.
`The present invention also provides a method of treating diabetes comprising
`administering to a patient in need of such treatment an effective amount of a shelf-stable
`pharmaceutical formulation, wherein the formulation comprises a therapeutically effective
`amount of a GLP-1 molecule, a pharmaceutically acceptable preservative, and a tonicity
`modifier, and wherein the formulation has a pH that is about 8.2 to about 8.8. In a
`preferred embodiment, the formulation used in the therapeutic method comprises a buffer,
`such as TRIS. In another preferred embodiment, the formulation used in the therapeutic
`method further comprises a surfactant, such as Brij-35. In an additional preferred
`embodiment, the GLP-1 moleculeof the formulation thus administered is an analog of GLP-
`1 and is selected from the group consisting of a peptide having the amino acid sequence:
`R1-X-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Y-Gly-Gln-Ala-Ala25-
`Lys-Z-Phe-Ile-Ala30-Trp-Leu-Val-Lys-Gly35-Arg-R2
`(SEQ ID N0:2)
`and a pharmaceutically-acceptable salt thereof, wherein R1 is His or desamino-histidine, X
`is Ala, Gly or Val, Y is Glu or Gln, Z is Glu or Gln and R2 is Gly-OH. In an especially
`preferred embodiment, the GLP-1 molecule administered is according to SEQ ID NO: 2,
`wherein R1 is L-histidine, Xis Val, Y is Glu, Z is Glu, and R2 is Gly-OH. In an alternative
`preferred embodiment, the GLP-1 molecule administered is a derivative of GLP-1 and is
`selected from the group consisting of a peptide having the amino acid sequence:
`NH2-His 7-Ala-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-V al-Ser-Ser-Tyr-Leu20-Glu-Gly-Gln-
`Ala-Ala25-Lys-Glu-Phe-Ile-Ala30-Trp-Leu-Val-X
`(SEQ ID N0:3)
`and a pharmaceutically-acceptable salt thereof, wherein X is selected from the group
`consisting of Lys and Lys-Gly; a pharmaceutically-acceptable lower alkylester of the peptide;
`and a pharmaceutically-acceptable amide of the peptide selected from the group consisting of
`amide, lower alkyl amide, and lower dialkyl amide.
`An additional embodiment encompasses a method of providing meal-time glycemic
`control and basal glycemic control with a single injection comprising administering to a
`patient in need thereof an effective amount of a shelf-stable pharmaceutical formulation,
`wherein the formulation comprises a therapeutically effective amount of a GLP-1 molecule,
`a long acting insulin agent, a pharmaceutically acceptable preservative, and a tonicity
`modifier, wherein said formulation has a pH that is about 8.2 to about 8.8.
`
`-5-
`
`MPI EXHIBIT 1074 PAGE 6
`
`MPI EXHIBIT 1074 PAGE 6
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0006
`
`
`
`WO 00/37098
`
`PCT/US99/30395
`
`captured in the pen which is disposed of after the contents of the cartridge have been
`exhausted.
`
`With the development of GLP-1 as well as analogs and derivatives thereof for the
`treatment of diabetes, there exists a need to improve treatment regimes that can balance
`chemical and physical stability with chronic use requirements of diabetic patients.
`
`Summary of the Invention
`In order to overcome the problems of chemical and physical stability of GLP-1
`formulations, the present inventors have developed a shelf-stable formulation of GLP-1. In
`particular, the inventors have discovered that when certain physiologically tolerated buffers
`are used in formulations of GLP-1, the physical stability of such formulations is
`unexpectedly and considerably greater than when compared to GLP-1 formulations made
`with a phosphate buffer. In addition, maintaining the pH in a range of about 8.2 to about
`8.8 unexpectedly improves the chemical stability of the formulations. The shelf-stable
`formulation of GLP-1 of the invention comprises a therapeutically effective amount of a
`GLP-1 molecule, a pharmaceutically acceptable preservative, and a tonicity modifier,
`wherein the pH of said formulation is maintained in the range from about 8.2 to about 8.8.
`In accordance with the chemical and physical stability needs of GLP-1 formulations,
`the present invention provides a shelf-stable pharmaceutical formulation comprising a
`therapeutically effective amount of a GLP-1 molecule, a pharmaceutically acceptable
`preservative, and a tonicity modifier, wherein the formulation has a pH that is about 8.2 to
`about 8. 8. In a preferred embodiment, the formulation further includes a buffer, such as
`TRIS. In another preferred embodiment, the formulation comprises a surfactant, such as
`Brij-35. In an additional preferred embodiment, the GLP-1 molecule of the formulation is
`an analog of GLP-1 and is selected from the group consisting of a peptide having the amino
`acid sequence:
`Ri-X-Glu-Gly 10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Y-Gly-Gln-Ala-Ala25-
`Lys-Z-Phe-Ile-Ala30-Trp-Leu-Val-Lys-Gly35-Arg-R2
`(SEQ ID N0:2)
`and a pharmaceutically-acceptable salt thereof, wherein R1 is His or desamino-histidine, X
`is Ala, Gly or Val, Y is Glu or Gln, Z is Glu or Gln and R2 is Gly-OH. In an especially
`preferred embodiement, the GLP-1 molecule is according to SEQ ID NO: 2, wherein R1 is L(cid:173)
`histidine, Xis Val, Y is Glu, Z is Glu, and R2 is Gly-OH. In an alternative preferred
`
`-3-
`
`MPI EXHIBIT 1074 PAGE 7
`
`MPI EXHIBIT 1074 PAGE 7
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0007
`
`
`
`WO 00/37098
`
`PCT/US99/30395
`
`Detailed Description of the Preferred Embodiments
`The present invention provides a shelf-stable formulation of GLP-1, GLP-1 analogs,
`and GLP-1 derivatives. Because it is known that there are problems with long term
`stability of GLP-1 as a component in a pharmaceutical composition, the present inventors
`developed a pharmaceutical formulation which stabilizes GLP-1, its derivatives and
`analogs. This development led to the shelf-stable GLP-1 formulations of the invention.
`In another embodiment, the present invention encompasses a GLP-1 formulation
`that also comprises a long acting diabetic agent. It has long been a goal of insulin therapy
`to mimic the pattern of endogenous insulin secretion in normal individuals. The daily
`physiological demand for insulin fluctuates and can be separated into two phases: (a) the
`absorptive phase requiring a pulse of insulin to dispose of the meal-related blood glucose
`surge, and (b) the post-absorptive phase requiring a sustained delivery of insulin to regulate
`hepatic glucose output for maintaining optimal fasting blood glucose. Accordingly,
`effective therapy for people with diabetes generally involves the combined use of two types
`of exogenous insulin formulations: a fast-acting meal time insulin provided by bolus
`injections and a long-acting, so-called, basal insulin, administered by injection once or
`twice daily to control blood glucose levels between meals. Examples of commercial basal
`insulin preparations include NPH (Neutral Protamine Hagedorn) insulin, protamine zinc
`insulin (PZI), and ultralente (UL).
`The term "stability" is used to mean chemical as well as physical stability. Physical
`stability refers to properties such as protein aggregation, which can be measured by a
`sample's attenuation of light. The measurement relates to the turbidity of a formulation.
`Turbidity is produced by aggregation or precipitation of proteins or complexes in the
`formulation and is indicative of decreased stability of a solution formulation. The more
`turbid a protein preparation, the less stable the preparation is. Stability also refers to the
`chemical stability of the formulation such as the propensity of the proteins to form high
`order polymers which is indicative of decreased stability.
`One factor that plays a role in the stability of GLP-1 formulations is the maintenance
`of pH at a prescribed level. Specifically, the present inventors have found that achieving
`and maintaining the pH of the formulation at about 8.2 to about 8.8 is advantageous.
`Typical peptide formulations have a more neutral pH of 7 to about 7. 8 or an acidic pH.
`Furthermore, a composition containing a GLP-1 molecule that has a pH in the range of
`
`-6-
`
`MPI EXHIBIT 1074 PAGE 8
`
`MPI EXHIBIT 1074 PAGE 8
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0008
`
`
`
`WO 00/37098
`
`PCT /US99/30395
`about 6.8 to about 7.5 exhibits less physical stability than a composition of a GLP-1
`molecule containing a preservative and having a pH in the range of about 8.2 to about 8.8.
`A preserved formulation which has a pH of less than about 8.0 tends to exhibit turbidity, a
`telltale sign of decreased physical stability of the peptide formulation. Conversely, a
`formulation which has a pH greater than about 8.8 tends to have decreased chemical
`stability.
`Therefore, the invention contemplates GLP-1 formulations having a pH range of
`about 8.2 to about 8.8, which preserves optimal chemical and physical stability of the GLP-
`1 molecule. A particularly preferred range for the inventive GLP-1 formulations is about
`8.3 to about 8.6, and a most particularly preferred pH range is about 8.4 to about 8.5. As
`used in this specification with respect to pH, the term "about" means plus or minus 0.1 pH
`units. Thus, a pH of "about 8.5" denotes a pH of 8.4 to 8.6.
`GLP-1 molecules themselves exhibit a buffering capacity. However, to maintain the
`pH of the composition for long term storage and stability, it is preferable to add a buffer.
`The choice of buffer affects the chemical and physical stability of the formulation
`because it influences pH. There are very few pharmaceutically acceptable buffers in the
`alkaline range. Phosphate buffers, which are typically used in peptide pharmaceutical
`formulations, cannot maintain a pH range of 8.2 to about 8.8. However, the present
`inventors have discovered that certain other amine-containing buffers are capable of
`imparting chemical as well as physical stability to formulations of GLP-1 molecules.
`The buffers used in the present invention preferably provide a buffering capacity in
`the range of about 8.2 to about 8.8. The buffers which are used may be tromethane
`(TRIS), and amino-acid based buffers such as lysine and hydroxy-lysine. Although any
`non-phosphate buffer which has a buffering capacity in the range of about 8.2 to about 8.8
`may be used, TRIS is the preferred buffer for the formulations of the present invention.
`The term "TRIS" refers to 2-amino-2-hydroxymethyl-1,3-propanediol ( also known in the
`art as tromethane, trimethylol aminomethane or tris(hydroxymethyl) aminomethane), and to
`any pharmacologically acceptable salt thereof. The free base and the hydrochloride form
`are two common forms of TRIS. TRIS is one of the few buffers which is capable of
`maintaining the pH at the alkaline level desired, thereby stabilizing the formulation.
`A second factor that plays a role in the stability of GLP-1 formulations is the
`concentration of the GLP-1 molecule that is used in the inventive formulation. The present
`
`-7-
`
`MPI EXHIBIT 1074 PAGE 9
`
`MPI EXHIBIT 1074 PAGE 9
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0009
`
`
`
`WO 00/37098
`
`PCT /US99/30395
`
`inventors determined that a concentration of about 0.30 to about 0.65 mg/ml of the GLP-1
`molecule was stable in the inventive formulations. However, a concentratioh which was
`about equal to or greater than 1 mg/ml was unstable. This stability was evidenced by the
`development of turbidity in the formulation. A particularly stable formulation includes
`about 0.5 mg/ml of a GLP-1 molecule.
`An additional factor which contributes to the overall stability of the GLP-1
`formulations of the present invention is the choice of preservative. The preservative is an
`essential component in the formulation because it enables multiple uses of the formulation.
`While it is typical that most preservatives would be capable of stabilizing a pharmaceutical
`formulation, some pharmaceutically acceptable preservatives act to promote physical
`instability of the formulation. The present inventors have found that a phenolic
`preservative is preferred. Specifically, the preservative may be m-cresol, phenol, benzyl
`alcohol, or methyl paraben and is present in an amount from about 2 mg/ml to about 6
`mg/ml. Ultimately, the concentration of preservative necessary for effective preservation
`depends on the preservative used, the pH of the formulation, and whether substances that
`bind or sequester the preservative are also present. Preferably, m-cresol is used in the
`formulations as a preservative.
`While a buffer and a preservative are most preferably included in the formulation,
`other additional excipients may be included, such as a tonicity modifier and/or a surfactant
`as well as distilled water for injections.
`The tonicity modifier may be included to make the formulation approximately
`isotonic with bodily fluid depending on the mode of administration. The concentration of
`the tonicity modifier is in accordance with the known concentration of a tonicity modifier in
`a peptide formulation. A preferable tonicity modifier used in the present invention is
`glycerol.
`A surfactant, which may be included in the formulation of the present invention, can
`be cationic, anionic, or non-ionic. A preferable class of surfactants is polyoxyethylene
`ethers. A preferred surfactant useful in the present invention is Brij-35, a polyoxyethylene
`23 lauryl ether, available from ICI United States, Inc.
`The present invention contemplates use of not only natural GLP-1, but also analogs,
`derivatives and salts of GLP-1. As used herein, the term "a GLP-1 molecule" refers (1) to
`the naturally-occurring GLP-1, which is GLP-1 (7-37)-OH; (2) GLP-1(7-36)NH2, as well
`
`-8-
`
`MPI EXHIBIT 1074 PAGE 10
`
`MPI EXHIBIT 1074 PAGE 10
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0010
`
`
`
`WO 00/37098
`
`PCT /US99/30395
`
`as (3) GLP-1 (7-37); (4) natural and synthetic functional GLP-1 analogs; (5) derivatives of
`
`GLP-1 and (6) salts of any of the aforementioned molecules.
`
`A "GLP-1 analog" is defined as a molecule having one or more amino acid
`
`substitutions, deletions, inversions, or additions relative to GLP-1(7-37) and may include the
`
`D-amino acid forms. Numerous GLP-1 analogs are known in the art and include, but are not
`limited to, GLP-1(7-34), GLP-1(7-35), GLP-1(7-36)NH2, Gln9-GLP-1(7-37), d-Gln9-GLP-
`1(7-37), Thr16-Lys18-GLP-1(7-37), and Lys18-GLP-1(7-37), Gly8-GLP-1(7-36)NH2, Gly8-GLP-
`1(7-37)0H, Val8-GLP-1(7-37)0H, Met8-GLP-1(7-37)0H, acetyl-Lys9-GLP-1(7-37), Thr9-
`GLP-1(7-37), D-Thr9-GLP-1(7-37), Asn9-GLP-1(7-37), D-Asn9-GLP-1(7-37), Ser-Arg23-
`Arg24-Gln26-GLP-1(7-37), Arg23-GLP-1(7-37), Arg24-GLP-1(7-37), a-methyl-Ala8-GLP-1(7-
`36)NH2, and Gly8-Gln21-GLP-1(7-37)0H, and the like.
`Other GLP-1 analogs consistent with the present invention are described by the
`
`formula:
`R1-X-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Y-Gly-Gln-Ala-Ala25-
`Lys-Z-Phe-Ile-Ala30-Trp-Leu-Val-Lys-Gly35-Arg-R2
`(SEQ ID N0:2)
`
`wherein: R1 is selected from the group consisting of L-histidine, D-histidine, desamino(cid:173)
`histidine, 2-amino-histidine, beta-hydroxy-histidine, homohistidine, alpha-fluoromethyl(cid:173)
`
`histidine, and alpha-methyl-histidine; X is selected from the group consisting of Ala, Gly,
`
`Val, Thr, Ile, and alpha-methyl-Ala; Y is selected from the group consisting of Glu, Gln,
`
`Ala, Thr, Ser, and Gly; Z is selected from the group consisting of Glu, Gln, Ala, Thr, Ser,
`and Gly; and R2 is selected from the group consisting of NH2, and Gly-OH.
`GLP-1 analogs also have been described in WO 91/11457, and include GLP-1(7-34),
`GLP-1(7-35), GLP-1(7-36), or GLP-1(7-37), or the amide form thereof, and
`
`pharmaceutically-acceptable salts thereof, having at least one modification selected from the
`
`group consisting of:
`
`(a) substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine,
`alanine, valine, isoleucine, leucine, methionine, phenylalanine, arginine, or D-lysine for
`
`lysine at position 26 and/or position 34; or substitution of glycine, serine, cysteine, threonine,
`asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine,
`
`phenylalanine, lysine, or a D-arginine for arginine at position 36;
`
`(b) substitution of an oxidation-resistant amino acid for tryptophan at position 31;
`
`MPI EXHIBIT 1074 PAGE 11
`
`MPI EXHIBIT 1074 PAGE 11
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0011
`
`
`
`PCT/US99/30395
`
`WO 00/37098
`(c) substitution of at least one of: tyrosine for valine at position 16; lysine for serine at
`position 18; aspartic acid for glutamic acid at position 21; serine for glycine at position 22;
`arginine for glutamine at position 23; arginine for alanine at position 24; and glutamine for
`lysine at position 26; and
`(d) substitution of at least one of: glycine, serine, or cysteine for alanine at position 8;
`aspartic acid, glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine,
`valine, isoleucine, leucine, methionine, or phenylalanine for glutamic acid at position 9;
`serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine,
`leucine, methionine, or phenylalanine for glycine at position 10; and glutamic acid for
`aspartic acid at position 15; and
`(e) substitution of glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine,
`alanine, valine, isoleucine, leucine, methionine, or phenylalanine, or the D- or N-acylated or
`alkylated form of histidine for histidine at position 7;
`wherein, in the substitutions described in (a), (b), (d), and (e), the substituted amino acids can
`optionally be in the D-form and the amino acids substituted at position 7 can optionally be in
`the N-acylated or N-alkylated form.
`Preferred GLP-1 molecules used in the present inventive formulation also include
`analogs of GLP-1 (7-37)NH2 and GLP-1 (7-37) in which one or more amino acids which are
`not present in the original sequence are added or deleted, and derivatives thereof.
`Specifically, His and desamino-histidine are preferred for R1• Ala, Gly and Val are preferred
`at the "X" position. Also, Glu and Gln are preferred for at the "Y" position. Glu and Gln
`are preferred at the "Z" position and Gly-OH is preferred for R2.
`A particularly preferred GLP-1 analog is known as Val(8)GLP-1 (VSGLP-1) and has
`a formula according to SEQ ID N0:2, wherein R1 is L-histidine, Xis Val, Y is Glu, Z is Glu
`and R2 is Gly-OH.
`A "GLP-1 derivative" is defined as a molecule having the amino acid sequence of
`GLP-1(7-37) or of a GLP-1 analog, but additionally comprises chemical modification of one
`or more of its amino acid side groups, a-carbon atoms, terminal amino group, or terminal
`carboxylic acid group. A chemical modification includes, but is not limited to, adding
`chemical moieties, creating new bonds, and removing chemical moieties. Modifications at
`amino acid side groups include, without limitation, acylation of lysine e-amino groups, N(cid:173)
`alkylation of arginine, histidine, or lysine, alkylation of glutamic or aspartic carboxylic acid
`
`-10-
`
`MPI EXHIBIT 1074 PAGE 12
`
`MPI EXHIBIT 1074 PAGE 12
`
`Apotex v. Novo - IPR2024-00631
`Petitioner Apotex Exhibit 1074-0012
`
`
`
`WO 00/37098
`
`PCT /US99/30395
`groups, and deamiCU4tion of glutamine or asparagine. Modifications of the tenninal amino
`include, without limitation, the des-amino, N-lower alkyl, N-di-lower alkyl, and N-acyl
`modifications. Modifications of the terminal car boxy group include, without limitation, the
`amide, lower alkyl amide, dialkyl amide, and lower alkyl ester modifications. Lower alkyl is
`C1-C4 alkyl. Furthermore, one or more side groups, or tenninal groups, may be protected by
`protective groups known to the ordinarily-skilled protein chemist The a-carbon of an amino
`acid maybe mono- or dimethylated.
`Other GLP-1 derivatives include molecules which are selected from the group
`consisting of a peptide having the amino acid sequence:
`
`NH2-His7-Ala-Glu-Gly10-Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-Glu-Gly-
`Gln-Ala-Ala25-Lys-Glu-Phe-Ile-Ala30-Trp-Leu-Val-X
`(SEQ ID N0:3)
`and pharmaceutically-acceptable salts thereof, wherein X is selected from the group consisting of
`Lys and Lys-Gly; and a derivative of said peptide, wherein said peptide is selected from the
`group consisting of: a pharmaceutically-acceptable lower alkylester of said peptide; and a
`pharmaceutically-acceptable amide of said peptide, selected from the group consisting of amide,
`lower alkyl amide, and lower dialkyl amide.
`Yet