Pharmaceutical compositions of glp-1 / glp-2 dual agonists

Abstract

The present invention relates to pharmaceutical compositions comprising specific preservatives.

Description

TECHNICAL FIELD

[0001] The present invention relates to pharmaceutical compositions comprising specific preservatives. The pharmaceutical compositions according to the present invention are particularly stable and have an advantageous shelf life. BACKGROUND

[0002] Peptides are an important part of the pharmaceutical industry. Despite great advances in the production of active pharmaceutical ingredients (APIs), the production of peptide-based drug products remains a significant challenge. Challenges associated with peptide formulation development are often overlooked or ignored.

[0003] Generally, peptides are defined as polypeptides of less than 50 amino acid residues and often lack organized tertiary or globular structure. Some do adopt secondary structure, although this tendency is limited, for example, single turn alpha-helices. While their smaller size makes them easier to deliver across biological barriers than larger proteins, their formulation can be difficult.

[0004] Some formulation challenges associated with peptides specifically include: chemical instability; adoption of multiple conformers; their tendency to self-associate; and complex physical instability, for example, gel formation, amyloid formation, and / or precipitation.

[0005] The most common challenge is chemical degradation of peptides and proteins, through degradation mechanisms such as isomerization, racemization, hydrolysis, deamidation, and oxidation. The amino acid sequence of a given peptide defines the extent to which it is affected by deamidation and / or oxidation reactions. The rate of oxidation of specific residues, for example, Met residues, is related to the degree of solvent exposure. Since peptides do not have a globular structure with chelatable reactive groups, the side chains of nearly all residues in a peptide are fully solvent-exposed to allow maximum contact with reactive oxygen species. Deamidation involves the hydrolysis of the amide side chain of amino acid residues such as Asn and Gin. Furthermore, the high degree of peptide chain flexibility results in a high deamidation rate compared to more complex proteins. It is important, however, to note that the nature of the amino acid after deamidation, for example, the nature of the amino acid after Asn, also affects the rate of deamidation. Peptides that lack steric bulk and the ability to form hydrogen bonds with the Asn side chain can even accelerate the reaction. Generally, Asn-Gly, Asn-Ala, Asn-Ser, and Asn-Asp amino acid combinations exhibit reaction rates that scientists must consider and test to ensure stable pharmaceutical compositions. The greatest control over hydrolytic reactions, including deamidation, is achieved by stable and reliable pH and buffer systems. However, such stable and reliable pH and buffer systems will be affected by additional excipients added to the composition.

[0006] Excipients are added to pharmaceutical compositions to enhance or maintain the solubility (solubilizers) and / or stability (buffers, antioxidants, chelating agents, cryoprotectants, and lyoprotectants) of the active ingredients. Excipients are important in parenteral formulations in many cases to ensure safety (antimicrobial preservatives), to minimize pain and irritation after injection (tonicity agents), and to control or prolong drug delivery (polymers). These are all examples of positive or synergistic interactions between excipients and the drug product. However, any excipient added to a composition has the potential to produce a negative effect, such as loss of peptide solubility, activity, and / or chemical / physical stability, increased self- aggregation or fibrillation, which in turn can make administration of the drug product unsafe.

[0007] Preservatives can be added to pharmaceutical compositions to kill microbial contaminants that can be introduced into the composition, such as when multiple aliquots are used or withdrawn from a container holding multiple doses of the drug. Pharmaceutical compositions can be sealed and stored under sterile conditions in the absence of a preservative, but when a container holding the composition is used, any accidentally introduced microorganisms can render the contents unsuitable for further medical use. Thus, it is important to effectively preserve the drug contents, especially when the composition is stored in bulk for several administrations. If a container holding a bulk amount of the unpreserved pharmaceutical composition is used, the lack of a preservative can mean that most of the contents are wasted. Preservatives advantageously enable the pharmaceutical composition to be stored at low temperatures (e.g., refrigerated at about 5°C) for months or years, or at higher temperatures (e.g., room temperature) for shorter periods of time, such as days or weeks, even after use of a portion of the composition.

[0008] However, while the use of preservatives has advantages, the inclusion of a preservative in a pharmaceutical composition can be problematic because the preservative can interact adversely with other components of the composition, particularly the active component. Such interactions can result in a reduction in the preservative action of the pharmaceutical composition, or a reduction or complete lack of drug efficacy. For example, the preservative can cause chemical instability of the active substance. In the case of a peptide having activity, the preservative can participate in or promote a degradation reaction, such as isomerization, racemization, hydrolysis, deamidation, or oxidation of the peptide, resulting in loss of pharmacological activity of the peptide. In addition or alternatively, the preservative can be detrimental to the physical stability of the active peptide, enhancing aggregation of the peptide into inactive covalent oligomers and / or causing the peptide to precipitate out of solution. Such loss of physical stability not only reduces the drug potency of the peptide, but the formation of particulate matter has practical and safety implications if the composition is delivered by injection.

[0009] Given the vast sequence diversity of peptides, and thus different chemical structures, it is inherently unpredictable whether a given substance will act as an effective preservative for a particular therapeutic peptide composition without negatively affecting the peptide in the above-described manner.

[0010] The present invention relates to pharmaceutical compositions comprising selected peptides disclosed in WO2018104561 (e.g. compound 18 of WO2018104561), which describes said compounds in detail and their use. SUMMARY

[0012] The present application provides compositions comprising one or more GLP-1 / GLP-2 dual agonist peptides and one or more preservatives in a buffer. In some aspects, the compositions are isotonic parenteral pharmaceutical compositions suitable for administration to a human subject.

[0013] GLP-1 / GLP-2 dual agonist peptides are peptides. Specific preservatives have been identified for use in compositions comprising specific GLP-1 / GLP-2 dual agonist peptides and specific buffers without substantially affecting the chemical or physical stability of the peptides. It was surprisingly found that specific preservatives have no effect on chemical stability at all peptide concentrations and only a slight effect on peptide oligomerization in phosphate buffers at higher peptide concentrations. Furthermore, it has been found that, counterintuitively, peptides oligomerize less at higher peptide concentrations when peptide compositions comprising m-cresol or phenol are stored for a short period (2 weeks) at high temperature (40°C) compared to at lower concentrations.

[0014] Thus, the compositions of the present invention benefit from the advantages associated with preservatives, i.e. prevention or reduction of microbial contamination.

[0015] It is particularly advantageous that the preservative effect in the compositions of the present invention allows the provision of the compositions in a multi-dose administration setting. The compositions of the present invention can be provided in a device for the continuous treatment of a dose of the composition at intervals over an extended period of time. During this period, the preservative effect prevents the growth of microorganisms in the composition while maintaining the chemical and physical stability of the peptides. The practical benefit is that the device only needs to be loaded once without the need to prepare a new dose for each administration.

[0016] In one aspect, the pharmaceutical compositions according to the present invention are for or suitable for administration in a multi-dose device.

[0017] In some aspects, the present invention provides a composition comprising:

[0018] (a) one or more GLP-1 / GLP-2 dual agonist peptides comprising the following general formula A:

[0019] H[Aib]EG-X5-F-X7-SELATILD-[ψ]-QAARDFIAWLI-X28-HKITD (A),

[0020] wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and at least one of X5 and X7 is T,

[0021] wherein [Ψ] represents an L or D lysine residue, wherein the albumin binding moiety is conjugated to the GLP-1 / GLP-2 dual agonist at this point, and

[0022] wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)];

[0023] (b) one or more preservatives,

[0024] wherein the one or more preservatives comprise or are meta-cresol and / or phenol; and

[0025] (c) a phosphate buffer.

[0026] In some aspects, specific and particular compositions, such as isotonic parenteral compositions, are described in detail in the specification of the present invention.

[0027] The chemical stability of the GLP-1 / GLP-2 dual agonist in any of the test compositions disclosed herein at a time point Y can be determined in terms of the relative purity X of the GLP-1 / GLP-2 dual agonist Y at a given time point Y, and by measuring the absolute purity X’ of the GLP-1 / GLP-2 dual agonist and normalizing it to the absolute purity X 0 of the GLP-1 / GLP-2 dual agonist at day zero (day 0), wherein the absolute purity is determined by HPLC at a given time point Y by identifying the purity of the peak corresponding to the GLP-1 / GLP-2 dual agonist.

[0028] Surprisingly, it was found that the GLP-1 / GLP-2 dual agonist in the disclosed compositions comprising a phosphate buffer and a preservative selected from meta-cresol and phenol has good chemical stability at all peptide concentrations. Furthermore, it was found that when the GLP-1 / GLP-2 dual agonist is stored with meta-cresol or phenol at high temperature (40°C) for a short period of time (2 weeks), contrary to intuition, the peptide oligomerizes less at higher peptide concentrations than at lower concentrations.

[0029] The present invention also provides a composition according to the present invention for use in:

[0030] (i) increasing intestinal mass, improving intestinal function, increasing intestinal blood flow, or repairing intestinal damage or dysfunction in a subject in need thereof; or

[0031] (ii) preventing or treating malabsorption, ulcer, short bowel syndrome, cul-de-sac syndrome, inflammatory bowel disease, irritable bowel syndrome, pouchitis, celiac sprue, tropical sprue, hypogammaglobulinemic sprue, mucositis induced by chemotherapy or radiation therapy, diarrhea induced by chemotherapy or radiation therapy, low grade inflammation, metabolic endotoxemia, necrotizing enterocolitis, primary biliary cirrhosis, hepatitis, fatty liver disease, or gastrointestinal side effects of inflammatory disorders in a subject in need thereof; or

[0032] (iii) reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss in a subject in need thereof; or

[0033] (iv) preventing or treating obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose intolerance, dyslipidemia, diabetes, prediabetes, metabolic syndrome, or hypertension in a subject in need thereof.

[0034] The present application also provides a method for preserving a composition comprising one or more GLP-1 / GLP-2 dual agonists of the present application and a phosphate buffer, wherein the method comprises adding one or more preservatives to the composition, wherein the one or more preservatives comprise or are meta-cresol and / or phenol.

[0035] The present application also provides the use of a preservative for preserving a composition comprising one or more GLP-1 / GLP-2 dual agonists of the present application and a phosphate buffer, wherein the preservative comprises or is meta-cresol and / or phenol. DETAILED DESCRIPTION

[0037] Compound

[0038] The compositions of the present application comprise one or more GLP-1 / GLP-2 dual agonists comprising the following general formula A:

[0039] H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD(A), wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and at least one of X5 and X7 is T, wherein [Ψ] represents an L or D lysine residue, wherein an albumin binding moiety is conjugated here to the GLP-1 / GLP-2 dual agonist, and wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)].

[0040] In some aspects, one or more GLP-1 / GLP-2 dual agonists comprising Formula A have Formula B:

[0041] H[Aib]EG-X5-FT-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD(B),

[0042] wherein X5 is T or S, X28 is Q, E, A, H, Y, L, K, R or S, wherein [Ψ] represents an L or D lysine residue, wherein an albumin binding moiety is conjugated here to the GLP-1 / GLP-2 dual agonist, and wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)].

[0043] In some aspects, one or more GLP-1 / GLP-2 dual agonists comprising Formula A or B comprise the sequence: H[Aib]EGSFTSELATILD[Ψ]QAARDFIAWLIQHKITD (SEQ ID NO: 1). In some aspects, one or more GLP-1 / GLP-2 dual agonists comprising Formula A or B consist of the sequence: H[Aib]EGSFTSELATILD[Ψ]QAARDFIAWLIQHKITD (SEQ ID NO: 1).

[0044] In some aspects, one or more GLP-1 / GLP-2 dual agonists comprising Formula A is: Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (CPD1OH) or any pharmaceutically acceptable salt thereof. In some aspects, the pharmaceutically acceptable salt of CPD1OH is a sodium salt, a hydrochloride salt, or an acetate salt, preferably a hydrochloride salt.

[0045] In some aspects, the one or more GLP-1 / GLP-2 dual agonist comprising general formula A is: Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-NH2 (CPD1NH2) or any pharmaceutically acceptable salt thereof. In some aspects, the pharmaceutically acceptable salt of CPD1NH2 is a sodium salt, a hydrochloride salt, or an acetate salt, preferably a hydrochloride salt.

[0046] In a preferred aspect, the one or more GLP-1 / GLP-2 dual agonist is CPD1OH or any pharmaceutically acceptable salt thereof, preferably a hydrochloride salt thereof.

[0047] As used herein, the term GLP-1 / GLP-2 dual agonist refers to a peptide that has activity at both GLP-1 receptors and GLP-2 receptors. The GLP-1 / GLP-2 dual agonist comprising formula A or formula B can be the peptide of SEQ ID NO: 1 or a peptide in which one or more amino acids are modified relative to SEQ ID NO: 1. Such agonists and / or peptides can also comprise one or more side chains that have been covalently attached to the GLP-1 / GLP-2 dual agonist. The term “side chain” can also be referred to as a “substituent.”

[0048] As used herein, the term “salt” refers to ionic compounds that can be formed by the neutralization reaction of acids and bases. Salts are composed of an associated number of cations (positively charged ions) and anions (negative ions) such that the product is electrically neutral (no net charge). These component ions can be inorganic, such as chloride (Cl - ), or organic, such as acetate (CH3CO 2- ); and can be monatomic, such as fluoride (F - ), or polyatomic, such as sulfate (SO4 2- ).

[0049] The term "pharmaceutically acceptable salt of CPD1" or "salt of CPD1" as used herein describes a salt of the compound comprising SEQ ID NO: 1. As used herein "Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]- isoGlu)]QAARDFIAWLIQHKITD-OH[acid]" refers to a salt of Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH, wherein [acid] refers to the acid that forms the salt of the compound in a neutralization reaction, for example Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH.[HC1]. Thus [HC1] would refer to the hydrochloride salt.

[0050] As used herein, "pharmaceutically acceptable salt" means a salt that is safe and effective for use in mammals and has the desired biological activity. Pharmaceutically acceptable salts include salts of acidic or basic groups present in CPD1. For a review on pharmaceutically acceptable salts see Berge et al., 66 J. Pharm. Sci. 1-19 (1977), which is incorporated herein by reference.

[0051] Table 1 - Selected GLP-1 / GLP-2 dual agonists included in the compositions of the present invention

[0052]

[0053] The abbreviation CPD1 refers to the compound comprising SEQ ID NO: 1 in any form. However, CPD1OH discloses only the compound comprising SEQ ID NO: 1, wherein the compound is in its -OH form (free acid). The CPD1NH2 form refers to the -NH2 form of the compound (amidated form). Both CPD1OH and CPD1NH2 can be converted to a pharmaceutically acceptable salt to provide a drug substance in powder form.

[0054] Table 2 - Amino acid sequences included in one or more of the GLP-1 / GLP-2 dual agonists of the present invention

[0055]

[0056] An "albumin binding moiety" facilitates circulation of the GLP-1 / GLP-2 dual agonist in the bloodstream and also has the effect of prolonging the action time of the GLP-1 / GLP-2 dual agonist. The albumin binding moiety binds the GLP-1 / GLP-2 dual agonist to albumin present in the blood and, due to the fact that the GLP-1 / GLP-2 dual agonist is released slowly only from the albumin, the action of the GLP-1 / GLP-2 dual agonist is prolonged. The term "albumin binding moiety" can also be referred to as a "side chain" or "substituent".

[0057] The term "natural amino acid" when used herein is an amino acid selected from the following (typically indicated in the three letter code in brackets and the one letter code): glycine (Gly and G), proline (Pro and P), alanine (Ala and A), valine (Val and V), leucine (Leu and L), isoleucine (Ile and I), methionine (Met and M), cysteine (Cys and C), phenylalanine (Phe and F), tyrosine (Tyr and Y), tryptophan (Trp and W), histidine (His and H), lysine (Lys and K), arginine (Arg and R), glutamine (Gln and Q), asparagine (Asn and N), glutamic acid (Glu and E), aspartic acid (Asp and D), serine (Ser and S) and threonine (Thr and T). If not otherwise indicated, an amino acid indicated in the one letter code in capital letters indicates the L-isoform, whereas if the amino acid is indicated in lower case, the amino acid is used / applied in its D-form, e.g. K (i.e. L-lysine), k (i.e. D-lysine).

[0058] The abbreviation "Hy-" in connection with the compounds disclosed herein means hydrogen. The abbreviation was chosen to avoid confusion of hydrogen with histidine (H) at the beginning of the sequence.

[0059] Throughout the specification and claims, other generally accepted three letter codes for "alpha-amino acids" are used, e.g. sarcosine (Sar), norleucine (Nle), alpha-aminoisobutyric acid (Aib), 2,3-diaminopropionic acid (Dap), 2,4-diaminobutyric acid (Dab) and 2,5-diaminopentanoic acid (ornithine; Orn). Such other alpha-amino acids can be indicated in square brackets "[ ]" (e.g. "[Aib]") when used in a general formula or sequence in the present specification, especially when the remainder of the formula or sequence is shown using one letter codes.

[0060] Concentration of compound

[0061] In some aspects, a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, comprises at least about 1 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 2 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, comprises at least about 1 mg / mL to about 33 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 2 mg / mL to about 33 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application comprises at least about 1 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 2 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 4 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 6 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 8 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist, e.g., at least about 10 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist.

[0062] In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 1 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 2 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 4 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 6 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 8 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 10 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 15 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 20 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 25 mg / mL of a GLP-1 / GLP-2 dual agonist. In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise about 33 mg / mL of a GLP-1 / GLP-2 dual agonist.

[0063] Preferably, the compositions of the present application comprise about 6 mg / mL to about 25 mg / mL of a GLP-1 / GLP-2 dual agonist, or about 2 mg / mL to about 10 mg / mL of a GLP-1 / GLP-2 dual agonist. Most preferably, the compositions of the present application comprise about 15 mg / mL of a GLP-1 / GLP-2 dual agonist.

[0064] Synthesis of dual agonists

[0065] The dual agonists of the application are preferably synthesized by solid phase or liquid phase peptide synthesis methods. In this context, reference can be made to WO 98 / 1 125 and in particular to Fields, G. B. et al., 2002, "Principles and practice of solid-phase peptide synthesis". In: Synthetic Peptides (2nd Edition) and the examples herein. According to the present application, the dual agonists of the application can be synthesized or produced in a variety of ways, including for example methods comprising:

[0066] (a) synthesizing a dual agonist by solid phase or liquid phase peptide synthesis and recovering the synthesized dual agonist thus obtained; or

[0067] (b) expressing a precursor peptide sequence from a nucleic acid construct encoding the precursor peptide, recovering the expression product, and modifying the precursor peptide to produce a compound of the application.

[0068] The precursor peptide can be modified by introducing one or more non-proteinogenic amino acids (e.g. Aib, Orn, Dap or Dab), introducing an albumin binding moiety or introducing an appropriate terminal group -OH or -NH2, etc.

[0069] Expression is typically carried out from a nucleic acid encoding the precursor peptide, which can be in a cell or cell-free expression system comprising such a nucleic acid.

[0070] Preservatives

[0071] The compositions of the application, e.g. the isotonic parenteral pharmaceutical compositions of the application, comprise one or more preservatives. In some aspects, the one or more preservatives comprise or are meta-cresol and / or phenol. In some aspects, the preservative is meta-cresol or phenol. The compositions of the application, e.g. the isotonic parenteral pharmaceutical compositions of the application, comprise a preservative, wherein the preservative comprises or is meta-cresol and / or phenol. In some aspects, the compositions of the application, e.g. the isotonic parenteral pharmaceutical compositions of the application, comprise one preservative, wherein the preservative comprises or is meta-cresol or phenol. In some aspects, the compositions of the application, e.g. the isotonic parenteral pharmaceutical compositions of the application, comprise two preservatives, wherein the preservatives comprise or are meta-cresol and phenol.

[0072] M-Cresol

[0073] In some aspects, the composition of the present application, e.g., the isotonic parenteral pharmaceutical composition of the present application, comprises one or more preservatives, wherein the one or more preservatives comprises meta-cresol. In some aspects, the one or more preservatives is meta-cresol. In some aspects, the composition of the present application, e.g., the isotonic parenteral pharmaceutical composition of the present application, comprises a preservative that is meta-cresol. In some aspects, the composition of the present application, e.g., the isotonic parenteral pharmaceutical composition of the present application, comprises meta-cresol.

[0074] Meta-cresol is an organic compound that is also known as meta-cresol, 3- cresol, 3-methylphenol, 3-methylbenzenol, 3-hydroxytoluene, or 1-hydroxy-3- methylbenzene. Meta-cresol has the chemical formula CH3C6H4(OH) and the following structural formula:

[0075]

[0076] In some aspects, meta-cresol is present in the composition of the present application at a concentration of about 1.15 mg / mL to about 5.15 mg / mL. In some aspects, meta-cresol is present in the composition of the present application at a concentration of about 1.15 mg / mL. In some aspects, meta-cresol is present in the composition of the present application at a concentration of about 5.15 mg / mL. Preferably, meta-cresol is present in the composition of the present application at a concentration of about 3.15 mg / mL.

[0077] Phenol

[0078] In some aspects, the composition of the present application, e.g., the isotonic parenteral pharmaceutical composition of the present application, comprises one or more preservatives, wherein the one or more preservatives comprises phenol. In some aspects, the one or more preservatives is phenol. In some aspects, the composition of the present application, e.g., the isotonic parenteral pharmaceutical composition of the present application, comprises a preservative that is phenol. In some aspects, the composition of the present application, e.g., the isotonic parenteral pharmaceutical composition of the present application, comprises phenol.

[0079] Phenol is an organic compound that is also known as benzenol. Phenol has the chemical formula C6H5OH and the following structural formula:

[0080]

[0081] In some aspects, the phenol is present in the compositions of the present application at a concentration of about 2.5 mg / mL to about 8.5 mg / mL. In some aspects, the phenol is present in the compositions of the present application at a concentration of about 2.5 mg / mL. In some aspects, the phenol is present in the compositions of the present application at a concentration of about 8.5 mg / mL. Preferably, the phenol is present in the compositions of the present application at a concentration of about 5.5 mg / mL.

[0082] Phosphate buffer and pH

[0083] The compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise a phosphate buffer.

[0084] In some aspects, the phosphate buffer is present in the composition, e.g., isotonic parenteral pharmaceutical composition, at a final concentration of about 5 mM to about 50 mM, e.g., about 5 mM to about 40 mM, e.g., about 5 mM to about 30 mM. Preferably, the phosphate buffer is present in the composition at a final concentration of about 5 mM to about 20 mM. In some aspects, the phosphate buffer is present in the composition at a final concentration of about 5 mM. In some aspects, the phosphate buffer is present in the composition at a final concentration of about 50 mM. Most preferably, the phosphate buffer is present in the composition at a final concentration of about 20 mM.

[0085] In some aspects, the phosphate buffer is a sodium phosphate buffer. In some aspects, the phosphate buffer is disodium hydrogen phosphate (Na2HP04) or sodium dihydrogen phosphate (NaH2P04), or a combination thereof.

[0086] In one aspect, the disodium hydrogen phosphate is present in the composition at a final concentration of about 15 mM to about 19 mM, preferably 18 mM to 19 mM.

[0087] In one aspect, the sodium dihydrogen phosphate is present in the isotonic parenteral pharmaceutical composition at a final concentration of about 1 mM to about 3 mM, preferably 1 mM to 2 mM.

[0088] In one aspect, the final concentration of the disodium hydrogen phosphate and sodium dihydrogen phosphate buffer components together is about 5 mM to about 50 mM, preferably about 10 mM to about 40 mM, more preferably about 15 mM to about 30 mM. In a most preferred aspect, the final concentration of the disodium hydrogen phosphate and sodium dihydrogen phosphate buffer components together is about 20 mM.

[0089] In some aspects, the pH of a composition of the application, e.g., an isotonic parenteral pharmaceutical composition, is about pH 6.0 to about pH 8.5, e.g., about pH 6.0 to about pH 8.4, about pH 6.0 to about pH 8.3, about pH 6.0 to about pH 8.2, about pH 6.0 to about pH 8.1, or about pH 6.0 to about pH 8.0. In some aspects, the pH is about pH 6.5 to about pH 8.5. In one aspect, the pH is preferably about pH 7.0 to about pH 8.0. In some aspects, the pH of a composition of the application, e.g., an isotonic parenteral pharmaceutical composition, is about pH 7.0 to about pH 8.0. In some aspects, the pH of a composition of the application, e.g., an isotonic parenteral pharmaceutical composition, is about pH 7.0. In some aspects, the pH of a composition of the application, e.g., an isotonic parenteral pharmaceutical composition, is about pH 8.0. In some aspects, the pH of a composition of the application is about pH 8.2. In some aspects, the pH of a composition of the application is about pH 6.0. In some aspects, the pH of a composition of the application is about pH 7.0 to about pH 8.2, preferably about pH 7.5 or about pH 8.2. In some aspects, the pH of a composition of the application is about pH 7.0 to about pH 8.2, preferably about pH 7.6 or about pH 8.0. In some aspects, the pH of a composition of the application is about pH 7.0 to about pH 8.2, preferably about pH 7.6 or about pH 7.7. In some aspects, the pH of a composition of the application is about pH 7.0 to about pH 8.2, preferably about pH 7.6. In some aspects, the pH of a composition of the application is about pH 7.0 to about pH 8.2, preferably about pH 8.0. In some aspects, the pH of a composition of the application is about pH 7.0 to about pH 8.2, preferably about pH 7.0. In a preferred aspect, the pH is about 8.0.

[0090] In some aspects, the pH in a composition of the application is adjusted as needed with NaOH or HC1.

[0091] Tonicity and tonicity agents

[0092] In some aspects, a composition of the application is an isotonic parenteral pharmaceutical composition.

[0093] In some aspects, a composition of the application comprising one or more GLP-1 / GLP-2 dual agonist comprising Formula A or Formula B is isotonic. In some aspects, a composition of the application comprising one or more GLP-1 / GLP-2 dual agonist comprising SEQ ID NO: 1 is isotonic.

[0094] In some embodiments, the osmolality of the compositions of the present application is about 300 ± 120 mOsmol / kg. In some embodiments, the osmolality of the compositions of the present application is about 290 ± 70 mOsmol / kg. In some embodiments, the osmolality of the compositions of the present application is about 230 mOsmol / kg to about 370 mOsmol / kg. In some embodiments, the osmolality of the compositions of the present application is about 280 mOsmol / kg to about 320 mOsmol / kg. In some embodiments, the osmolality of the compositions of the present application is about 290 mOsmol / kg to about 320 mOsmol / kg.

[0095] In some aspects, the compositions of the present application, e.g., the isotonic parenteral pharmaceutical compositions of the present application, comprise one or more tonicity agents.

[0096] The term "tonicity agent" refers to an excipient added to the compositions according to the present application to achieve isotonicity with respect to body fluids. A range of ionic and non-ionic tonicity agents are used in pharmaceutical compositions. Non-ionic tonicity agents can be selected from the group consisting of dextrose, propylene glycol, glyceryl, mannitol, e.g., D-mannitol, and sorbitol. Ionic tonicity agents can include alkali or alkaline earth metal halides, e.g., CaCl2, KBr, KC1, LiCl, Nal, NaBr, NaCl, Na2SO4. In one aspect, the tonicity agent can be selected from the group consisting of mannitol, sodium chloride, and propylene glycol.

[0097] An "ionic compound" is two or more ions held together by attractive forces. An example of an ionic compound is table salt. Table salt is composed of positive sodium ions and negative chloride ions. It has a high melting and boiling point and is hard or brittle. It is also soluble in water. The definition of a "non-ionic compound" is that the chemical bonds in the compound are non-ionic. They typically have chemical bonds that share electron density.

[0098] In some aspects, the one or more tonicity agents comprise or are mannitol. Preferably, the one or more tonicity agents is D-mannitol. In some aspects, the mannitol, e.g., D-mannitol, is present in the compositions of the present application at a concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, more preferably about 190 mM to about 240 mM. In some aspects, the mannitol, e.g., D-mannitol, is present in the compositions of the present application at about 230 mM.

[0099] In some aspects, the one or more tonicity agents comprise or are NaCl. In some aspects, the NaCl is present in the compositions of the present application at a concentration of about 50 mM to about 450 mM, preferably about 65 mM to about 165 mM. Preferably, the NaCl is present at a concentration of about 125 mM.

[0100] The term "isotonic" as used herein refers to the tonicity at the site of injection (i.e. i.v. or s.c.) relative to body fluids. Thus, the term "isotonic" is used to describe a pharmaceutical composition that has the same tonicity at the site of injection as body fluids (e.g. red blood cells and / or plasma). Compositions having an osmolarity of about 300 mOsmol / kg, for example about 280 to 320 mOsmol / kg or about 290 to 320 mOsmol / kg are considered isotonic.

[0101] Isotonicity is important for parenteral pharmaceutical compositions because "hypotonic" solutions cause cells to swell, while "hypertonic" solutions cause cells to shrink. Although it is related to osmolarity, tonicity also takes into account the ability of solutes to cross cell membranes.

[0102] Compositions of the present invention

[0103] In some aspects, the compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, comprise a solvent. In some aspects, the solvent is water.

[0104] In some aspects, the compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, comprise a tonicity agent and a solvent.

[0105] In some aspects, the compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, comprise a GLP-1 / GLP-2 dual agonist comprising the amino acid sequence of Formula A, m-cresol, and a phosphate buffer.

[0106] In some aspects, the compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, comprise a GLP-1 / GLP-2 dual agonist comprising the amino acid sequence of Formula A, phenol, and a phosphate buffer.

[0107] In some aspects, the compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, comprise a GLP-1 / GLP-2 dual agonist comprising the amino acid sequence of Formula A, m-cresol, a phosphate buffer, and mannitol, e.g. D-mannitol.

[0108] In some aspects, the compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, comprise a GLP-1 / GLP-2 dual agonist comprising the amino acid sequence of Formula A, phenol, a phosphate buffer, and mannitol, e.g. D-mannitol.

[0109] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0110] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0111] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A, the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0112] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A and is present at a concentration of about 1 mg / mL to about 33 mg / mL, preferably about 1 mg / mL to about 25 mg / mL, preferably about 6 mg / mL to about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, or the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0113] In some aspects, in a composition of the present invention, e.g., an isotonic parenteral pharmaceutical composition of the present invention, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A and is present at a concentration of about 1 mg / mL to about 33 mg / mL, preferably at a concentration of about 1 mg / mL to about 25 mg / mL, preferably about 6 mg / mL to about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0114] In some aspects, in a composition of the present invention, e.g., an isotonic parenteral pharmaceutical composition of the present invention, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A and is present at a concentration of about 1 mg / mL to about 33 mg / mL, preferably at a concentration of about 1 mg / mL to about 25 mg / mL, preferably about 6 mg / mL to about 25 mg / mL, the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0115] In some aspects, in a composition of the present invention, e.g., an isotonic parenteral pharmaceutical composition of the present invention, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A and is present at a concentration of about 1 mg / mL to about 33 mg / mL, preferably at a concentration of about 1 mg / mL to about 25 mg / mL, preferably about 6 mg / mL to about 25 mg / mL, more preferably about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, or the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM. The composition preferably further comprises mannitol, e.g., D-mannitol, at a concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0116] In some aspects, in a composition of the application, e.g., an isotonic parenteral pharmaceutical composition of the application, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A and is present at a concentration of about 1 mg / mL to about 33 mg / mL, preferably at a concentration of about 1 mg / mL to about 25 mg / mL, preferably about 6 mg / mL to about 25 mg / mL, more preferably about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM. The composition preferably further comprises mannitol, e.g., D-mannitol, at a concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0117] In some aspects, in a composition of the application, e.g., an isotonic parenteral pharmaceutical composition of the application, the GLP-1 / GLP-2 dual agonist comprises an amino acid sequence of Formula A and is present at a concentration of about 1 mg / mL to about 33 mg / mL, preferably at a concentration of about 1 mg / mL to about 25 mg / mL, preferably about 6 mg / mL to about 25 mg / mL, more preferably about 25 mg / mL, the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM. The composition preferably further comprises mannitol, e.g., D-mannitol, at a concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0118] In some aspects, in a composition of the application, e.g., an isotonic parenteral pharmaceutical composition of the application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof at a final concentration of about 1 mg / mL to about 33 mg / mL, preferably about 1 mg / mL to about 25 mg / mL, preferably about 2 mg / mL to about 25 mg / mL, more preferably about 6 mg / mL to about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, or the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0119] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, preferably at a final concentration of about 1 mg / mL to about 33 mg / mL, preferably about 1 mg / mL to about 25 mg / mL, preferably about 2 mg / mL to about 25 mg / mL, more preferably about 6 mg / mL to about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0120] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, preferably at a final concentration of about 1 mg / mL to about 33 mg / mL, preferably about 1 mg / mL to about 25 mg / mL, preferably about 2 mg / mL to about 25 mg / mL, more preferably about 6 mg / mL to about 25 mg / mL, the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, and the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM.

[0121] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0122] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0123] In some aspects, in a composition of the application, e.g., an isotonic parenteral pharmaceutical composition of the application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, preferably at a final concentration of about 1 mg / mL to about 33 mg / mL, preferably about 1 mg / mL to about 25 mg / mL, preferably about 2 mg / mL to about 25 mg / mL, more preferably about 6 mg / mL to about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, or the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0124] In some aspects, in a composition of the application, e.g., an isotonic parenteral pharmaceutical composition of the application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, preferably at a final concentration of about 1 mg / mL to about 33 mg / mL, preferably about 1 mg / mL to about 25 mg / mL, preferably about 2 mg / mL to about 25 mg / mL, more preferably about 6 mg / mL to about 25 mg / mL, the preservative is m-cresol at a final concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0125] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, the preservative is phenol at a final concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, the phosphate buffer is at a final concentration of about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0126] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, the preservative is m-cresol at a final concentration of about 3.15 mg / mL, the phosphate buffer is at a final concentration of about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 230 mM.

[0127] In some aspects, in a composition of the present application, e.g., an isotonic parenteral pharmaceutical composition of the present application, the GLP-1 / GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, the preservative is phenol at a final concentration of about 5.5 mg / mL, the phosphate buffer is at a final concentration of about 20 mM, and the tonicity agent is mannitol, e.g., D-mannitol, at a final concentration of about 230 mM.

[0128] Indications

[0129] In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of prevention or treatment of intestinal injury and dysfunction, modulation of body weight, and prevention or treatment of metabolic dysfunction.

[0130] In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of prevention or treatment of malabsorption, ulcers (e.g., peptic ulcers, Zollinger-Ellison Syndrome, drug-induced ulcers, and ulcers associated with infection or other pathogens), short bowel syndrome, blind loop syndrome, inflammatory bowel disease (Crohns disease and ulcerative colitis), irritable bowel syndrome (IBS), pouchitis, celiac sprue (e.g., induced by gluten or celiac disease), tropical sprue, hypogammaglobulinemic sprue, mucositis induced by chemotherapy or radiation therapy, diarrhea induced by chemotherapy or radiation therapy, low-grade inflammation, metabolic endotoxemia, necrotizing enterocolitis, primary biliary cirrhosis, hepatitis, fatty liver disease (including parenteral nutrition-associated intestinal atrophy, PNALD (Parenteral Nutrition-Associated Liver Disease), NAFLD (Non-Alcoholic Fatty Liver Disease), and NASH (Non-Alcoholic Steatohepatitis)), or gastrointestinal side effects of inflammatory conditions (e.g., pancreatitis) or graft versus host disease (GVHD).

[0131] In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of prevention or treatment of obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose intolerance, dyslipidemia (e.g., elevated LDL levels or reduced HDL / LDL ratio), diabetes (e.g., type 2 diabetes, gestational diabetes), prediabetes, metabolic syndrome, or hypertension.

[0132] In some aspects, the pharmaceutical composition of the present application is administered to a human subject to promote a biological effect selected from the group consisting of increasing intestinal mass, improving intestinal function (especially intestinal barrier function), increasing intestinal blood flow, repairing intestinal damage or dysfunction, in a subject in need thereof.

[0133] In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of prevention or treatment of intestinal dysfunction or damage caused by or associated with GVHD, and prevention or treatment of side effects such as diarrhea caused by or associated with GVHD.

[0134] In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of prevention or treatment of obesity, morbid obesity, obesity-related gallbladder disease, and obesity-induced sleep apnea.

[0135] In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of improvement of glucose tolerance and / or glucose control. In some aspects, the pharmaceutical composition of the present application is administered to a human subject in need of modulation (e.g., improvement) of circulating cholesterol levels, reduction of circulating triglyceride or LDL levels, and increase of HDL / LDL ratio.

[0136] Administration

[0137] In some aspects, the pharmaceutical composition of the present application is an aqueous composition. In some aspects, the pharmaceutical composition of the present application is suitable for parenteral administration by injection, optionally by pen-like injection, subcutaneously (s.c.), intramuscularly (i.m.), or intravenously (i.v.) with the aid of a syringe. In some aspects, the pharmaceutical composition of the present application is suitable for s.c. injection into a human subject. In some aspects, the pharmaceutical composition of the present application is suitable for (i.v.) injection into a human subject.

[0138] In some aspects, the isotonic parenteral pharmaceutical composition of the present application is suitable for single dose administration. In some aspects, the injected isotonic parenteral pharmaceutical composition is suitable for injection with a single-use device. In some aspects, the single-use device is selected from a syringe pen or a single-use syringe. In some aspects, the isotonic parenteral pharmaceutical composition of the present application is suitable for multiple dose administration.

[0139] In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 1 mg to about 25 mg. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 1 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 2 mg to about 25 mg, preferably in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 6 mg to about 25 mg. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 1 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 2 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 3 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 4 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 5 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 6 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 9 mg or more. In some aspects, the isotonic parenteral pharmaceutical compositions of the present application are administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 10 mg or more.In some aspects, the isotonic parenteral pharmaceutical composition of the present application is administered to a human subject by s.c. injection in a volume that allows a total amount of GLP-1 / GLP-2 dual agonist to be delivered to the subject of about 7, 8, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 mg or more.

[0140] Biological activity

[0141] In some aspects, the peptide comprised in the pharmaceutical composition of the present application is the peptide according to Formula A and SEQ ID NO: 1, which has previously been described in patent application WO2018104561, which describes the compound, its preparation and purification, and biological activity (Table 5, WO2018104561). Example 2 in WO2018104561 includes data on in vitro potency on GLP-1 and GLP-2 receptors.

[0142] Chemical stability: purity

[0143] The compositions of the present application, e.g. the isotonic parenteral pharmaceutical compositions of the present application, provide good chemical stability. In other words, in the compositions of the present application, the GLP-1 / GLP-2 dual agonist remains chemically stable during storage. The chemical stability can be comparable to, or improved compared to, an equivalent composition not comprising a preservative according to the present application as described herein.

[0144] In some aspects, the compositions of the present application have good relative purity. The relative purity can be comparable to, or improved compared to, an equivalent composition not comprising a preservative according to the present application as described herein.

[0145] When reference is made herein to the “chemical stability” of a composition of the present application, this means the chemical stability of the GLP-1 / GLP-2 dual agonist comprised in the composition. In some aspects, the chemical stability of the GLP-1 / GLP-2 dual agonist is determined using Assay I described herein.

[0146] The chemical stability of the GLP-1 / GLP-2 dual agonist at time point Y in any of the test compositions disclosed herein can be expressed as the relative purity X of the GLP-1 / GLP-2 dual agonist at time point Y Y and by measuring the absolute purity X’ of the GLP-1 / GLP-2 dual agonist and normalizing it to the absolute purity X 0 of the GLP-1 / GLP-2 dual agonist at day zero (day 0), wherein said absolute purity is determined by HPLC at a given time point Y by identifying the purity of the peak corresponding to the GLP-1 / GLP-2 dual agonist.

[0147] Thus, on day zero (day 0), the absolute purity X’ is identical to the absolute purity X 0 and thus the chemical stability of the GLP-1 / GLP-2 dual agonist in the test composition is expressed in relative purity X Y = 100% where Y = day 0.

[0148] The relative purity can be calculated by:

[0149] X Y = (X’ / X 0 ) * 100

[0150] where X is the relative purity at a given time point Y, X 0 is the absolute purity at day 0, and X’ is the absolute purity at a given time point Y,

[0151] where the absolute purity X 0 or X’ of the GLP-1 / GLP-2 dual agonist in the test composition is determined by HPLC identifying the purity of the peak corresponding to the GLP-1 / GLP-2 dual agonist.

[0152] It was surprisingly found that high peptide concentrations have no or little impact on the degradation of the peptide (chemical stability) in the presence of the preservatives m-cresol and phenol compared to low peptide concentrations.

[0153] The relative purity at a given time point can be calculated by multiplying the purity slope by the number of weeks of storage and subtracting the modulus of this value from 100%.

[0154] In some aspects, the pharmaceutical compositions of the present application result in a relative purity of at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the one or more GLP-1 / GLP-2 dual agonists (e.g., CPD1 or any pharmaceutically acceptable salt thereof) after, for example, storage at, for example, 40°C for at least 2 weeks. In some aspects, the pharmaceutical compositions of the present application result in a relative purity of at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the one or more GLP-1 / GLP-2 dual agonists (e.g., CPD1 or any pharmaceutically acceptable salt thereof) after, for example, storage at, for example, 25°C for at least 26 weeks. In some aspects, the pharmaceutical compositions of the present application result in a relative purity of at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the one or more GLP-1 / GLP-2 dual agonists (e.g., CPD1 or any pharmaceutically acceptable salt thereof) after, for example, storage at, for example, 5°C for at least 52 weeks.

[0155] Chemical stability: oligomerization

[0156] Peptides in solution can aggregate to form covalently linked oligomers that are therapeutically inactive. Peptide oligomerization can be measured using size exclusion chromatography (SEC), for example as described in Assay II herein.

[0157] A variety of analytical techniques are available in the art for measuring peptide stability and are reviewed, for example, in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993).

[0158] Surprisingly, it was found that high peptide concentrations in the presence of the preservatives m-cresol and phenol reduce the amount of covalent oligomers compared to low peptide concentrations and that a plateau effect exists at peptide concentrations above 6 mg / mL.

[0159] The relative total non-oligomerized peptide at a given time point can be calculated by multiplying the oligomer slope by the number of weeks of storage and subtracting the modulus of this value from 100%.

[0160] In some aspects, the pharmaceutical compositions of the present application result in at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% relative total non-oligomerized peptide (monomer) of the one or more GLP-1 / GLP-2 dual agonist (e.g., CPD1, or any pharmaceutically acceptable salt thereof) after, for example, storage at, for example, 40 °C for at least 2 weeks. In some aspects, the pharmaceutical compositions of the present application result in at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% relative total non-oligomerized peptide of the one or more GLP-1 / GLP-2 dual agonist (e.g., CPD1, or any pharmaceutically acceptable salt thereof) after, for example, storage at, for example, 25 °C for at least 26 weeks. In some aspects, the pharmaceutical compositions of the present application result in at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% relative total non-oligomerized peptide of the one or more GLP-1 / GLP-2 dual agonist (e.g., CPD1, or any pharmaceutically acceptable salt thereof) after, for example, storage at, for example, 5 °C for at least 52 weeks.

[0161] List of Abbreviations

[0162] Abbreviations Explanation i.v. intravenous s.c. subcutaneous HPLC high performance liquid chromatography ND not determined N.P. no particles part / cont particles / container RP-HPLC reverse phase high performance liquid chromatography CPD compound SEQ ID NO sequence identification number

[0163] Terms and Definitions

[0164] When terms related to numerical values such as "about" and "approximately" are used, the skilled artisan will immediately recognize that any action or result associated with a given value can be obtained within a certain tolerance of the particular value. Thus, the term "about" as used herein means reasonably close to the stated numerical value, e.g., plus or minus 10%. When the term "about" is used in the present patent application with respect to chemical stability, the reasonably close will be less than 2%, e.g., 0.5% or 0.75%, 1% or 1.5%.

[0165] The term "solvent" as used herein means a substance that dissolves a solute (a chemically different liquid, solid, or gas) to produce a solution. Solvents are usually liquids, but can also be solids, gases, or supercritical fluids. Solvents are often classified by polarity and are considered polar or nonpolar, as indicated by the dielectric constant. Generally, solvents with a dielectric constant greater than about 5 are considered "polar" and solvents with a dielectric constant less than 5 are considered "nonpolar."

[0166] Some non-limiting aspects of the present application

[0167] The following sections of the specification contain some particular, non-limiting aspects of the application. The aspects described below can be combined with any of the aspects of the application described above and below and herein:

[0168] 1. A composition comprising:

[0169] (a) one or more GLP-1 / GLP-2 dual agonists comprising the following general formula A:

[0170] H[Aib]EG-X5-F-X7-SELATILD-[ψ]-QAARDFIAWLI-X28-HKITD (A),

[0171] wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S; and at least one of X5 and X7 is T,

[0172] wherein [Ψ] represents an L or D lysine residue, wherein an albumin binding moiety is conjugated here to the GLP-1 / GLP-2 dual agonist, and

[0173] wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)],

[0174] (b) one or more preservatives, wherein the one or more preservatives comprise or are m-cresol and / or phenol; and

[0175] (c) a phosphate buffer.

[0176] 2. The composition according to aspect 1, wherein the composition is an isotonic parenteral pharmaceutical composition.

[0177] 3. The composition according to any one of the preceding aspects, wherein the one or more preservatives comprise or are m-cresol, preferably wherein the m-cresol is present at a concentration of about 1.15 mg / mL to about 5.15 mg / mL, more preferably wherein the m-cresol is present at a concentration of about 3.15 mg / mL.

[0178] 4. The composition according to any one of the preceding aspects, wherein the one or more preservatives comprise or are phenol, preferably wherein the phenol is present at a concentration of about 2.5 mg / mL to about 8.5 mg / mL, more preferably wherein the phenol is present at a concentration of about 5.5 mg / mL.

[0179] 5. The composition according to any one of the preceding aspects, wherein the phosphate buffer is present at a concentration of about 5 mM to about 50 mM, preferably wherein the phosphate buffer is present at a concentration of about 20 mM.

[0180] 6. The composition according to any one of the preceding aspects, wherein the phosphate buffer is a sodium phosphate buffer, preferably wherein the sodium phosphate buffer is selected from disodium hydrogen phosphate or sodium dihydrogen phosphate or a combination thereof.

[0181] 7. The composition according to any one of the preceding aspects, wherein the pH of the composition is from about pH 6.0 to about pH 8.5, preferably from about pH 6.5 to about pH 8.5, preferably the pH is from about pH 7.0 to about pH 8.0, more preferably the pH is about pH 8.0.

[0182] 8. The composition according to any one of the preceding aspects, wherein the one or more GLP-1 / GLP-2 dual agonist has the general formula B:

[0183] H[Aib]EG-X5-FT-SELATILD-[ψ]-QAARDFIAWLI-X28-HKITD (B),

[0184] wherein X5 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and

[0185] wherein [ψ] represents an L or D lysine residue, wherein an albumin binding moiety is conjugated to the GLP-1 / GLP-2 dual agonist at this point, and

[0186] wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)].

[0187] 9. The composition according to any one of the preceding aspects, wherein the one or more GLP-1 / GLP-2 dual agonist comprises the following sequence:

[0188] H[Aib]EGSFTSELATILD[ψ]QAARDFIAWLIQHKITD (SEQ ID NO: 1).

[0189] 10. The composition according to any one of the preceding aspects, wherein the one or more GLP-1 / GLP-2 dual agonist is:

[0190] Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (CPD1 OH); or

[0191] Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-NH2(CPD1NH2),

[0192] or a pharmaceutically acceptable salt of CPD1OH or CPD1NH2, preferably a hydrochloride salt of CPD1OH or CPD1NH2.

[0193] 11. The composition according to any one of the preceding aspects, wherein the GLP-1 / GLP-2 dual agonist is present at a concentration of at least about 1 mg / mL, preferably at a concentration of about 1 mg / mL to about 33 mg / mL, for example at a concentration of about 2 mg / mL to about 33 mg / mL, for example at a concentration of about 1 mg / mL to about 25 mg / mL, for example at a concentration of about 6 mg / mL to about 25 mg / mL.

[0194] 12. The composition according to aspect 11, wherein the GLP-1 / GLP-2 dual agonist is present at a concentration of about 2 mg / mL, about 15 mg / mL, or about 25 mg / mL.

[0195] 13. The composition according to any one of the preceding aspects, wherein the composition further comprises one or more tonicity agents.

[0196] 14. The composition according to aspect 13, wherein the one or more tonicity agents comprise mannitol, preferably D-mannitol, or are mannitol, preferably D-mannitol.

[0197] 15. The composition according to aspect 14, wherein the mannitol is present at a concentration of about 130 mM to about 330 mM, preferably at a concentration of about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

[0198] 16. The composition according to aspect 13, wherein the one or more tonicity agents comprise NaCl or are NaCl, preferably wherein the NaCl is present at a concentration of about 50 mM to about 450 mM, preferably at a concentration of about 65 mM to about 165 mM, preferably about 125 mM.

[0199] 17. The composition according to any one of the preceding aspects, wherein the osmolality of the composition is about 230 mOsmol / kg to about 370 mOsmol / kg.

[0200] 18. The composition according to any one of the preceding aspects, wherein the composition further comprises a solvent, preferably water.

[0201] 19. The composition according to any one of the preceding aspects, wherein the one or more preservatives is m-cresol at a concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, wherein the phosphate buffer is at a concentration of about 5 mM to about 50 mM, preferably about 20 mM, wherein the composition further comprises mannitol, preferably D-mannitol, at a concentration of about 130 mM to about 330 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM, and wherein the pH of the composition is about pH 7.0 to about pH 8.0, preferably about pH 8.0.

[0202] 20. The composition according to any one of the preceding aspects, wherein the one or more preservatives is phenol at a concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, wherein the phosphate buffer is at a concentration of about 5 mM to about 50 mM, preferably about 20 mM, wherein the composition further comprises mannitol, preferably D-mannitol, at a concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM, and wherein the pH of the composition is about pH 6.0 to about pH 8.5, preferably about pH 7.0 to about pH 8.0, preferably about pH 8.0.

[0203] 21. The composition according to any one of the preceding aspects, wherein the one or more GLP-1 / GLP-2 dual agonist is CPD1OH or CPD1NH2, preferably CPD1OH, or a hydrochloride salt of CPD1OH or CPD1NH2, preferably a hydrochloride salt of CPD1OH, wherein the one or more preservative is m-cresol at a concentration of about 1.15 mg / mL to about 5.15 mg / mL, preferably about 3.15 mg / mL, wherein the phosphate buffer is at a concentration of about 5 mM to about 50 mM, preferably about 20 mM, and wherein the composition further comprises mannitol, preferably D-mannitol, at a concentration of about 230 mM, water, and sodium hydroxide and / or hydrochloric acid to adjust the pH to about pH 8.0.

[0204] 22. The composition according to any one of the preceding aspects, wherein the one or more GLP-1 / GLP-2 dual agonist is CPD1OH or CPD1NH2, preferably CPD1OH, or a hydrochloride salt of CPD1OH or CPD1NH2, preferably a hydrochloride salt of CPD1OH, wherein the one or more preservative is phenol at a concentration of about 2.5 mg / mL to about 8.5 mg / mL, preferably about 5.5 mg / mL, wherein the phosphate buffer is at a concentration of about 5 mM to about 50 mM, preferably about 20 mM, and wherein the composition further comprises mannitol, preferably D-mannitol, at a concentration of about 230 mM, water, and sodium hydroxide and / or hydrochloric acid to adjust the pH to about pH 8.0.

[0205] 23. The composition according to any one of the preceding aspects, wherein the composition is suitable for subcutaneous (s.c.) or intravenous (i.v.) injection into a human subject.

[0206] 24. The composition according to any one of the preceding aspects, for use in:

[0207] (i) increasing intestinal mass, improving intestinal function, increasing intestinal blood flow, or repairing intestinal injury or dysfunction in a subject in need thereof; or

[0208] (ii) preventing or treating malabsorption, ulcer, short bowel syndrome, blind loop syndrome, inflammatory bowel disease, irritable bowel syndrome, pouchitis, celiac sprue, tropical sprue, hypogammaglobulinemic sprue, chemotherapy- or radiation-induced mucositis, chemotherapy- or radiation-induced diarrhea, low-grade inflammation, metabolic endotoxemia, necrotizing enterocolitis, primary biliary cirrhosis, hepatitis, fatty liver disease, or gastrointestinal side effects of inflammatory conditions in a subject in need thereof; or

[0209] (iii) reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss in a subject in need thereof; or

[0210] (iv) preventing or treating obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose intolerance, dyslipidemia, diabetes, prediabetes, metabolic syndrome, or hypertension in a subject in need thereof.

[0211] 25. A method for preserving a composition comprising one or more GLP-1 / GLP-2 dual agonist comprising the following general formula A:

[0212] H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD(A),

[0213] wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S; and at least one of X5 and X7 is T, and

[0214] wherein [Ψ] represents an L or D lysine residue, wherein an albumin binding moiety is conjugated to said GLP-1 / GLP-2 dual agonist at this point, and

[0215] wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)];

[0216] wherein the composition comprises a phosphate buffer; and

[0217] wherein the method comprises adding one or more preservatives to the composition, wherein the one or more preservatives comprise or are meta-cresol and / or phenol.

[0218] 26. Use of a preservative for preserving a composition comprising one or more GLP-1 / GLP-2 dual agonists comprising the following general formula A:

[0219] H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD(A),

[0220] wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S; and at least one of X5 and X7 is T, and

[0221] wherein [Ψ] represents an L or D lysine residue, wherein an albumin binding moiety is conjugated to said GLP-1 / GLP-2 dual agonist at this point, and

[0222] wherein the albumin binding moiety is [K([17-carboxy-heptadecanoyl]-isoGlu)];

[0223] wherein the composition comprises a phosphate buffer; and

[0224] wherein the preservative comprises or is meta-cresol and / or phenol.

[0225] General methods used

[0226] Method I - Method for preparing GLP-1 / GLP-2 dual agonists

[0227] GLP-1 / GLP-2 dual agonists were synthesized as described in general peptide synthesis and Example 1 of patent application WO2018 / 104561.

[0228] CPD1 (corresponding to compound 18 in WO2018 / 104561) was synthesized using the method of solid phase peptide synthesis (SPPS) and standard Fmoc coupling procedures. After completion of synthesis, the peptide sequence was deprotected and cleaved from the solid support and the crude peptide was purified using preparative reverse phase HPLC. The peptide was converted to an acceptable salt form (HCI, acetate or Na) and lyophilized to provide the final CPD1 drug substance.

[0229] Method II - Methods for preparing and analyzing pharmaceutical compositions

[0230] Sample solutions for laboratory scale

[0231] GLP-1 / GLP-2 dual agonist drug substance (CPD1) was prepared according to Method I and dissolved in MilliQ water (MQW) to give a stock solution of 40 mg / mL active pharmaceutical ingredient (API). The pH was measured. Subsequently the ingredients as exemplified in Tables 5 to 8 were added and mixed, and then the pH was adjusted as needed using 1 M NaOH / HCI to reach the appropriate pH. The final concentration of CPD1 was 2, 4, 6, 8, 10, 15 or 25 mg / mL as shown in the tables and examples in this application. Compositions for laboratory scale were prepared in a volume of 0.5 mL to about 2 mL.

[0232] For stability testing, samples were stored in a dark room (i.e. lights turned off) at 25 °C for 26 weeks or 52 weeks, or at 40 °C for 2 weeks (as shown in the examples). Samples were analyzed by RP-HPLC and SEC-HPLC according to Assay I and II, respectively.

[0233] Assay I - Measuring GLP-1 / GLP-2 dual agonist purity and determining % purity of normalized GLP-1 / GLP-2 dual agonists using RP-HPLC Table 4 - RP-HPLC method details

[0234] Chemical stability of GLP-1 / GLP-2 dual agonist (peptide) as prepared according to Method I comprised in parenteral pharmaceutical compositions as prepared according to Method II is analyzed according to the following method:

[0235] Chemical stability of GLP-1 / GLP-2 dual agonist (peptide) as prepared according to Method I comprised in parenteral pharmaceutical compositions as prepared according to Method II is analyzed according to the following method:

[0236] A Dionex Ultimate 3000 HPLC system (Thermo Fisher) was used with a flow rate of 0.5 mL / min giving a linear gradient for analysis. The mobile phase components consisted of 90% acetonitrile / 10% MQW with 0.3% trifluoroacetic acid (TFA) and 0.3% TFA in MQW. A wavelength of 215 nm was used for detection. The injection volume was 2 pg of peptide. The column used for HPLC analysis was a Phenomenex Kinetex C18, 150 x 3.0 mm, 2.6 pm particle size. The run time was 25 minutes.

[0237] Example 1 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol

[0238]

[0239] The results are shown in Tables 5 to 8 as degradation slopes calculated from the normalized purity results. The slope is a measure of how fast CPD1 degrades. Lower values (i.e. away from 0) represent higher degradation.

[0240] Method II - Method for evaluation of covalent oligomers by size exclusion chromatography (SEC)

[0241] Size exclusion chromatography (SEC) experiments were performed on a Dionex Ultimate 3000 HPLC system (ThermoFisher) using isocratic elution at a flow rate of 0.5 mL / min. The mobile phase consisted of 45% acetonitrile, 0.1% TFA in MQW. A wavelength of 215 nm was used for detection. The injection volume was 2 pg of peptide. The column used for SEC analysis was a TSKgel SuperSW2000 (TOSOH Corporation), 4 pm, 30 x 4.6 mm, and the column temperature was 25 °C. The run time was 12 minutes.

[0242] The SEC method is able to detect covalent oligomers, where two or more CPD1 molecules are linked together by covalent chemical bonds.

[0243] Oligomerization data is shown in Tables 5 to 8. Data is expressed as the slope calculated from the covalent oligomer results over time. The slope is a measure of how quickly CPD1 forms covalent oligomers. Higher numerical values represent higher covalent oligomer formation. Examples

[0244] These examples investigate the chemical stability and oligomerization of CPD1 in compositions containing different preservatives and buffers stored at different temperatures for different lengths of time.

[0245] CPD1 was produced according to Method I. Pharmaceutical compositions (i.e. formulations) containing different preservatives were prepared and stored according to Method II. The peptide is CPD1 OH, which comprises the amino acid sequence of Formula A. CPD1 OH can be interchanged with CPD1 NH2.

[0246] The chemical stability of CPD1 is expressed as the slope of the relative purity of the peptide over time. The slope is determined by measuring the absolute purity of the peptide peak (i.e. the CPD1 main peak) at each time point using HPLC as described in Assay I, then normalizing this value against the absolute purity of the peptide peak at T=0, which is set to 100% purity, to give the relative purity percentage at each time point. The slope is calculated from these relative purity values over the complete time course.

[0247] The oligomerization of CPD1 is expressed as the slope of the proportion of covalent oligomers over time, as determined using Assay II at each time point.

[0248] Example 2 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol stored at 40°C for 2 weeks

[0249] In this example, compositions (Formulations A to F) containing no preservative m-cresol or phenol and containing 2 mg / mL or 10 mg / mL peptide were stored at 25 °C for 52 weeks or at 5 °C for 52 weeks. The composition of Formulations A to F is shown in Table 5, as well as the chemical stability (purity) slope and oligomerization slope for each storage temperature and period.

[0250] The total proportion of chemically stable non-oligomerized CPD1 in each formulation after storage was calculated by subtracting the percentage of oligomerized peptide from the final percentage purity of the peptide. These results are shown in Table 6.

[0251] Example 3 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol and different peptide concentrations Table 5 - CPD1 purity and oligomerization slope for Formulations A-F

[0252] In this example, compositions containing different concentrations of m-cresol or phenol preservative and different concentrations of phosphate buffer were stored at 40°C for 2 weeks. The chemical stability (purity) slope and oligomerization slope for each formulation are shown in Table 7.

[0253] Table 6 - Percentage of chemically stable, non-oligomerized CPD1 in Formulations A-F Table 7 - CPD1 purity and oligomerization slope for Formulations 1-17

[0254] In this example, compositions containing m-cresol or phenol preservative and different concentrations of CDP1 peptide were stored at 40°C for 2 weeks. The chemical stability (purity) slope and oligomerization slope for each formulation are shown in Table 8.

[0255] The data show that high peptide concentrations reduced the amount of covalent oligomers in the presence of the preservatives m-cresol and phenol compared to low peptide concentrations (see Table 7, 2 mg / mL peptide vs. 25 mg / mL peptide), and there was a plateau effect at concentrations higher than 6 mg / mL (see Table 8, showing oligomerization of peptides at a concentration range of 2 mg / mL to 25 mg / mL). In general, the RP-HPLC purity data show that high peptide concentrations had little or no effect on peptide degradation.

[0256] All publications mentioned in the above specification are herein incorporated by reference. Various modifications and variations to the described methods and systems of the application will be apparent to those skilled in the art without departing from the scope and spirit of the application. Although the application has been described in connection with specific preferred embodiments, it should be understood that the application as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes of carrying out the application that are obvious to those skilled in biochemistry, molecular biology or related fields are intended to be within the scope of the following claims.

[0257] Table 8 - CPD1 purity and oligomerization slope for Formulations 18-31 (different peptide concentrations)

[0258]

[0259] Based on 13 week slope

[0260] Table 4 - RP-HPLC method details

[0261]

[0262] Example 1 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol

[0263]

[0264]

[0265] Example 2 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol stored at 40°C for 2 weeks Example 3 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol and different peptide concentrations Table 5 - CPD1 purity and oligomerization slope for Formulations A-F Table 6 - Percentage of chemically stable, non-oligomerized CPD1 in Formulations A-F Table 7 - CPD1 purity and oligomerization slope for Formulations 1-17 Table 8 - CPD1 purity and oligomerization slope for Formulations 18-31 (different peptide concentrations)

[0266]

[0267] The slope of the 2 to 10 mg / mL formulation was calculated based on 52-week data, and the slope of the 15 to 25 mg / mL samples was based on 26-week data. SEQUENCE LISTING <110> Zealand Pharma A / S <120> Pharmaceutical compositions of GLP-1 / GLP-2 dual agonists <130> P121098PCT <160> 3 <170> PatentIn version 3.5 <210> 1 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> GLP-1 / GLP-2 dual agonist peptide <220> <221> MOD_RES <222> (2)..(2) <223> Aib <220> <221> MOD_RES <222> (16)..(16) <223> Xaa is Lys or D-Lys, wherein the albumin binding moiety [K([17-carboxy- heptadecanoyl]-isoGlu)] is conjugated to the GLP-1 / GLP-2 dual agonist at this point <400> 1 His Xaa Glu Gly Ser Phe Thr Ser Glu Leu Ala Thr Ile Leu Asp Xaa 1 5 10 15 Gln Ala Ala Arg Asp Phe lie Ala Trp Leu lie Gin His Lys lie Thr 20 25 30 Asp <210> 2 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> GLP-1 / GLP-2 dual agonist peptide (Formula A) <220> <221> MOD_RES <222> (2)..(2) <223> Aib <220> <221> MISC_FEATURE <222> (5)..(5) <223> Xaa is Thr or Ser <220> <221> MISC_FEATURE <222> (7)..(7) <223> Xaa is Thr or Ser <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or D-Lys, wherein the albumin binding moiety [K([17-carboxy- heptadecanoyl]-isoGlu)] is conjugated to the GLP-1 / GLP-2 dual agonist at this point <220> <221> MISC_FEATURE <222> (28)..(28) <223> Xaa is Gin, Glu, Ala, His, Tyr, Leu, Lys, Arg, or Ser <400> 2 His Xaa Glu Gly Xaa Phe Xaa Ser Glu Leu Ala Thr lie Leu Asp Xaa 1 5 10 15 Gln Ala Ala Arg Asp Phe Ile Ala Trp Leu Ile Xaa His Lys Ile Thr 20 25 30 Asp <210> 3 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> GLP-1 / GLP-2 dual agonist peptide (Formula B) <220> <221> MOD_RES <222> (2)..(2) <223> Aib <220> <221> MISC_FEATURE <222> (5)..(5) <223> Xaa is Thr or Ser <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa is Lys or D-Lys, wherein the albumin binding moiety [K([17-carboxy- heptadecanoyl]-isoGlu)] is conjugated to the GLP-1 / GLP-2 dual agonist at this point <220> <221> MISC_FEATURE <222> (28)..(28) <223> Xaa is Gin, Glu, Ala, His, Tyr, Leu, Lys, Arg, or Ser <400> 3 His Xaa Glu Gly Xaa Phe Thr Ser Glu Leu Ala Thr Ile Leu Asp Xaa 1 5 10 15 Gln Ala Ala Arg Asp Phe Ile Ala Trp Leu Ile Xaa His Lys Ile Thr 20 25 30 Asp

Claims

1. A composition comprising: (a) one or more GLP-1 / GLP-2 dual agonists, wherein the one or more GLP-1 / GLP-2 dual agonists is Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl] isoGlu)]QAARDFIAWLIQHKITD-OH (CPD1 OH) or a pharmaceutically acceptable salt of CPD1 OH; (b) one or more preservatives, wherein the one or more preservatives comprises or is meta-cresol and / or phenol; and (c) a phosphate buffer.

2. The composition of claim 1, wherein the composition is an isotonic parenteral pharmaceutical composition.

3. The composition of claim 1, wherein the one or more preservatives comprises or is meta-cresol.

4. The composition of claim 3, wherein the meta-cresol is present at a concentration of 1.15 mg / mL to 5.15 mg / mL.

5. The composition of claim 3, wherein the meta-cresol is present at a concentration of 3.15 mg / mL.

6. The composition of claim 1, wherein the one or more preservatives comprises or is phenol.

7. The composition of claim 6, wherein the phenol is present at a concentration of 2.5 mg / mL to 8.5 mg / mL.

8. The composition of claim 6, wherein the phenol is present at a concentration of 5.5 mg / mL.

9. The composition of any one of claims 1 to 8, wherein the phosphate buffer is present at a concentration of 5 mM to 50 mM.

10. The composition of any one of claims 1 to 8, wherein the phosphate buffer is present at a concentration of 20 mM.

11. The composition of any one of claims 1 to 8, wherein the phosphate buffer is a sodium phosphate buffer.

12. The composition of claim 11, wherein the sodium phosphate buffer is selected from disodium hydrogen phosphate or sodium dihydrogen phosphate or a combination thereof.

13. The composition of any one of claims 1 to 8, wherein the pH of the composition is pH 6.0 to pH 8.

5.

14. The composition of claim 13, wherein the pH of the composition is pH 6.5 to pH 8.

5.

15. The composition of claim 13, wherein the pH of the composition is pH 7.0 to pH 8.

0.

16. The composition of claim 13, wherein the pH of the composition is pH 8.

0.

17. The composition of any one of claims 1 to 8, wherein the one or more GLP-1 / GLP-2 dual agonists is a hydrochloride salt of CPD1 OH.

18. The composition of any one of claims 1 to 8, wherein the one or more GLP-1 / GLP-2 dual agonists is present at a concentration of at least 1 mg / mL.

19. The composition of any one of claims 1-8, wherein the one or more GLP-l / GLP-2 dual agonists are present at a concentration of 1 mg / mL to 33 mg / mL.

20. The composition of any one of claims 1-8, wherein the one or more GLP-l / GLP-2 dual agonists are present at a concentration of 2 mg / mL to 33 mg / mL.

21. The composition of any one of claims 1-8, wherein the one or more GLP-l / GLP-2 dual agonists are present at a concentration of 6 mg / mL to 33 mg / mL.

22. The composition of any one of claims 1-8, wherein the one or more GLP-l / GLP-2 dual agonists are present at a concentration of 1 mg / mL to 25 mg / mL.

23. The composition of any one of claims 1-8, wherein the one or more GLP-l / GLP-2 dual agonists are present at a concentration of 6 mg / mL to 25 mg / mL.

24. The composition of claim 18, wherein the one or more GLP-l / GLP-2 dual agonists are present at a concentration of 2 mg / mL, 15 mg / mL, 25 mg / mL, or 33 mg / mL.

25. The composition of any one of claims 1-8, wherein the composition further comprises one or more tonicity agents.

26. The composition of claim 25, wherein the one or more tonicity agents comprise or are mannitol.

27. The composition of claim 26, wherein the mannitol is D-mannitol.

28. The composition of claim 26, wherein the mannitol is present at a concentration of 130 mM to 330 mM.

29. The composition of claim 26, wherein the mannitol is present at a concentration of 150 mM to 300 mM.

30. The composition of claim 26, wherein the mannitol is present at a concentration of 190 mM to 240 mM.

31. The composition of claim 26, wherein the mannitol is present at a concentration of 230 mM.

32. The composition of claim 25, wherein the one or more tonicity agents comprise or are NaCl.

33. The composition of claim 32, wherein the NaCl is present at a concentration of 50 mM to 450 mM.

34. The composition of claim 32, wherein the NaCl is present at a concentration of 65 mM to 165 mM.

35. The composition of claim 32, wherein the NaCl is present at a concentration of 125 mM.

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