TW202339788A - Pharmaceutical formulations comprising a cyclodextrin - Google Patents

Abstract

Disclosed herein is a liquid pharmaceutical formulation comprising an amylin receptor agonist, a GLP-1 receptor agonist and a cyclodextrin comprising hydroxypropyl substitutions. Said co-formulation may be used for the medical treatment of subjects with overweight or obesity, with or without associated co-morbidities; diabetes, with or without associated comorbidities; cardiovascular diseases, non-alcoholic steatohepatitis (NASH) and cognitive impairment, such as that caused by Alzheimer's disease.

Description

Pharmaceutical formulations including cyclodextrins

The present invention relates to a pharmaceutical formulation, which is a co-formulation of a GLP-1 receptor agonist and an amyloid receptor agonist. The pharmaceutical formulations may be used in the medical treatment of overweight or obesity with or without one or more associated comorbidities; diabetes with or without one or more associated comorbidities; cardiovascular disease; non-alcoholic steatotic hepatitis (NASH) ); and individuals with cognitive disabilities, such as those caused by Alzheimer's disease.

Semaglutide is a glucagon-like peptide 1 (GLP-1) receptor agonist and the active pharmaceutical ingredient in Ozempic®. Ozempic® is indicated (i) as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes and (ii) to reduce the risk of significant cardiovascular disease in adults with type 2 diabetes and established cardiovascular disease. Risk of adverse cardiovascular events.

Semaglutide is also the active pharmaceutical ingredient of Wegovy®. Ozempic® is indicated as an adjunct to a reduced calorie diet and increased physical activity in the presence of at least one weight-related comorbidity, in patients with an initial body mass index of 30 kg/m ² or above 27 kg/m ² of adult patients undergoing chronic weight management.

Ozempic® and Wegovy® are liquid pharmaceutical formulations including 8 mM phosphate and having a pH of approximately 7.4.

Fixed-dose combinations of the amyloid receptor agonist caglitide and the GLP-1 receptor agonist semaglutide have been studied for the treatment of overweight and obesity (Lancet 2021; 397: 1736–48). The drug products studied were in the form of individual liquid pharmaceutical formulations for subcutaneous use, which included caglitide or semaglutide, respectively.

To date, the possibility of coformulating semaglutide and caglitide has not been considered due to the different physicochemical properties of these active pharmaceutical ingredients. Semaglutide, a GLP-1 receptor agonist, has an isoelectric point that is incompatible with the pH-optimal caglitinide, an amyloid receptor agonist. Semaglutide is most stable at pH 7.4 and has previously required formulation in a neutral to slightly alkaline solution at pH 7 to 8 to ensure its solubility in aqueous solutions. Caglitinide is most stable at pH 4.0 and needs to be formulated in an acidic solution. Increasing the pH will accelerate the rate of chemical degradation. The different physicochemical properties of caglitide and semaglutide preclude a simple mixture of these two peptides. The same applies to other GLP-1 receptor agonist and amylin receptor agonist combinations when the two have incompatible optimal pH ranges.

There remains a need in the art for a simple means to co-administer a GLP-1 receptor agonist (such as semaglutide) and an amylin receptor agonist (such as caglitinide).

This article discloses a method of co-formulating amyloid receptor agonists and GLP-1 receptor agonists. Disclosed herein is a liquid pharmaceutical formulation that includes an amyloid receptor agonist, a GLP-1 receptor agonist, and a cyclodextrin that includes hydrophilic chemical substitution, such as hydroxypropyl substitution. The cyclodextrin system may be a hydroxypropyl-substituted alpha form, which includes six glucose units arranged in a ring. The cyclodextrin system may be a hydroxypropyl-substituted beta form, which includes seven glucose units arranged in a ring. The pharmaceutical formulation may also include a buffer (such as histidine), a tonicity agent (such as sorbitol), and/or a surfactant (such as polysorbate 80); and have a pH of about 5.5 to 6.5. The pharmaceutical formulations disclosed herein may be administered by parenteral injection, preferably subcutaneous injection.

The pharmaceutical formulations disclosed herein are useful in the medical treatment of individuals with: overweight or obesity with or without associated comorbidities; diabetes with or without associated comorbidities; cardiovascular disease; non-alcoholic steatotic hepatitis (NASH); and those with cognitive disabilities, such as those caused by Alzheimer's disease. The pharmaceutical formulations disclosed herein may improve convenience, treatment compliance, and ultimately clinical outcomes in patients.

The present invention is a liquid pharmaceutical preparation, which includes an amyloid receptor agonist, a GLP-1 receptor agonist and a hydroxypropyl-substituted cyclodextrin.

The pharmaceutical formulations disclosed herein include two active pharmaceutical ingredients, a GLP-1 receptor agonist and an amylin receptor agonist.

Disclosed herein is a method of co-formulating an amyloid receptor agonist and a GLP-1 receptor agonist, wherein the GLP-1 receptor agonist has an isoelectric point that excludes co-formulation within a pH range , thereby making the amyloid receptor agonist chemically stable. Disclosed herein is a means of co-formulating a GLP-1 receptor agonist with an isoelectric point (pI) below 6.0, such as 3.5 to 6.0, such as 4.0 to 6.0, with an amylin receptor agonist.

The optimal pH of an amyloid receptor agonist is the pH at which it is the most chemically and physically stable. The optimal pH of a GLP-1 receptor agonist is the pH at which it is chemically and physically most stable. The physical stability of a GLP-1 receptor agonist may reflect its isoelectric point, which may be consistent with the pH at which worst physical stability is expected.

Disclosed herein is a means of formulating an amyloid receptor agonist and a GLP-1 receptor agonist whose optimal pH values differ by at least about two pH units, such as 2 to 5 pH units, such as 2 to 4 pH units, such as 3 to 5 pH units.

The GLP-1 receptor agonist may be semaglutide. The amyloid receptor agonist may be caglitinide or a bioactive metabolite of caglitinide. The compositions of the formulations disclosed herein improve the chemical and physical stability of those active pharmaceutical ingredients; preserve the bioavailability and exposure pharmacokinetic profile of the active pharmaceutical ingredients; and exhibit acceptable performance when injected subcutaneously. Local tolerance.

The terms "pharmaceutical formulation," "co-formulation" and "drug product" are used interchangeably herein to mean a liquid pharmaceutical formulation that includes a GLP-1 receptor agonist and an amylin receptor agonist.

The pharmaceutical formulations disclosed herein are suitable for parenteral injection, preferably subcutaneous injection. Amylin

The term "amyloid" as used herein refers to a polypeptide having the same amino acid sequence as endogenous amylin, such as human amylin. amyloid receptor

Amylin compounds can target the calcitonin receptor (CTR) and/or the amyloid receptor (AMYR). The latter consists of an isoform of two components: the calcitonin receptor (CTR) and one of three receptor activity-modifying proteins (RAMP1–3) resulting in three possible complexes AMYR1–3. amyloid receptor agonist

The pharmaceutical formulations disclosed herein include an amylin receptor agonist. An "amyloid receptor agonist" can be defined as a chemical entity that binds to and activates the amyloid receptor. In the context of the present invention, an "amyloid receptor agonist" is at least capable of binding and activating amyloid receptor 3 (AMYR3). Amylin receptor agonists can also agonize calcitonin receptors and AMYR1-2.

Examples of endogenous amyloid receptor agonists are human amylin and human calcitonin. An example of an exogenous amyloid receptor agonist is caglitinide. An example of an exogenous amyloid receptor agonist is pramlintide.

The amyloid receptor agonist in the pharmaceutical formulations disclosed herein may be caglitinide or a bioactive metabolite of caglitinide.

The bioactive metabolite of caglitinide may have aspartate (Asp) at position 21 or 22. The bioactive metabolite of caglitinide may have isoaspartate (iso-Asp) at position 21 or 22. caglitinide

Caglitide is an amyloid receptor agonist also known as AM833. It is the compound of Example 53 in WO2012/168432: N-α-[(S)-4-carboxy-4-(19-carboxynonaderyl)butyl]-[Glu14, Arg17, Pro37]-pramlintide. Caglitinide can be prepared as described on pages 153 to 155 of WO2012/168432.

Caglitinide may be in the form of a salt, preferably a pharmaceutically acceptable salt.

The concentration of caglitinide in the pharmaceutical formulations disclosed herein can be from about 0.25 mg/ml to about 22 mg/ml.

Pharmaceutical formulations disclosed herein may include caglitinide at a concentration of about 0.33 to 18 mg/ml; such as 0.25 to 0.5 mg/ml, such as about 0.33 mg/ml; such as 0.5 to 1.0 mg/ml, such as about 0.67 mg/ml. ml; such as 1.0 to 1.5 mg/ml, such as about 1.33 mg/ml; such as 1.5 to 2.0 mg/ml, such as about 1.5 mg/ml; such as 2.0 to 2.5 mg/ml; such as 2.5 to 3.0 mg/ml; such as 3.0 to 3.5 mg/ml; such as about 3.2 mg/ml; such as 3.5 to 4.0 mg/ml; such as 4.0 to 5.0 mg/ml; such as 5.0 to 6.0 mg/ml; such as 6.0 to 7.0 mg/ml, such as 7.0 to 8.0 mg /ml, such as 8.0 to 9.0 mg/ml, such as 9.0 to 10.0 mg/ml, such as about 9.6 mg/ml; such as 10 to 11 mg/ml, such as 11.0 to 12.0 mg/ml, such as 11 to 13 mg/ml; Such as 13 to 22 mg/ml, such as about 18 mg/ml; such as about 20 to 22 mg/ml.

Pharmaceutical formulations disclosed herein may include no more than 22 mg/ml of caglitinide. Pharmaceutical formulations disclosed herein may include no more than 12 mg/ml of caglitinide. GLP-1

The term "GLP-1" or "native GLP-1" as used herein refers to human glucagon-like peptide-1 (GLP-1(7-37)). GLP-1 receptor agonists

The pharmaceutical formulations disclosed herein include GLP-1 receptor agonists. "GLP-1 receptor agonist" can be defined as a biological response that binds to the GLP-1 receptor and produces a response similar to the endogenous ligand glucagon-like peptide 1 (GLP-1(7-37)) ligand. "All" GLP-1 receptor agonists can be defined as GLP-1 receptor agonists that elicit a biological response of the same magnitude as GLP-1(7-37).

An example of an exogenous GLP-1 receptor agonist is semaglutide. An example of an exogenous GLP-1 receptor agonist is tilsiparatide. Semaglutide

Semaglutide is a GLP-1 receptor agonist, also known as N ^6.26- {18-[N-(17-carboxyheptadecanyl)-L-γ-glutaminyl]-10-side oxygen group -3,6,12,15-tetraoxa-9,18-diazaoctadecanoyl}-[8-(2-amino-2-propionic acid),34-L-arginine] human Glucagon-like peptide 1(7-37). Semaglutide can be prepared as described in Example 4 of WO2006/097537.

Semaglutide can be present in the composition in a fully or partially ionized form; for example, one or more carboxylic acid groups (-COOH) can be deprotonated to a carboxyl group (-COO ^- ) and/or one or more amine groups The group (-NH ₂ ) can be protonated to the -NH ₃ ⁺ group.

Semaglutide may be in the form of a salt, preferably a pharmaceutically acceptable salt.

The concentration of semaglutide in the pharmaceutical formulations disclosed herein can range from about 0.25 mg/ml to about 22 mg/ml.

The pharmaceutical formulation may include semaglutide at a concentration of about 0.33 to 18 mg/ml; such as 0.25 to 0.5 mg/ml, such as about 0.33 mg/ml; such as 0.5 to 1.0 mg/ml, such as about 0.67 mg/ml; such as 1.0 to 1.5 mg/ml, such as about 1.33 mg/ml; such as 1.5 to 2.0 mg/ml, such as about 1.5 mg/ml; such as 2.0 to 2.5 mg/ml; such as about 2.2 mg/ml; such as 2.5 to 3.0 mg/ml ml; such as 3.0 to 3.5 mg/ml; such as about 3.2 mg/ml; such as 3.5 to 4.0 mg/ml; such as 4.0 to 5.0 mg/ml; such as about 4.8 mg/ml; such as 5.0 to 6.0 mg/ml; such as 6.0 to 7.0 mg/ml, such as about 6.4 mg/ml; such as 7.0 to 8.0 mg/ml, such as about 8.0 mg/ml; such as 8.0 to 9.0 mg/ml, such as 9.0 to 10.0 mg/ml, such as about 9.6 mg/ml ; Such as 10 to 11 mg/ml, such as about 10.7 mg/ml; Such as 11.0 to 12.0 mg/ml, such as 11 to 13 mg/ml; Such as about 12.8 mg/ml; Such as 13 to 22 mg/ml, such as about 16 mg/ml; such as about 18 mg/ml; such as about 20 to 22 mg/ml.

Pharmaceutical formulations disclosed herein may include no more than 22 mg/ml of semaglutide. Pharmaceutical formulations disclosed herein may include no more than 12 mg/ml of semaglutide. isoelectric point

The isoelectric point (pI) of a molecule is the pH value at which the molecule has no net charge. The pI of a peptide can be calculated theoretically from the pK values of its amino acid and terminal amine and carboxyl groups and can be used to predict the solubility of the peptide at a given pH.

The theoretically calculated isoelectric point of a GLP-1 receptor agonist may be in the range of 3.5 to 6.0, such as 4.0 to 6.0, such as 3.8 to 4.9, such as 4.0 to 4.5. Semaglutide has a theoretically calculated isoelectric point of approximately 4.37.

Theoretically calculated isoelectric points of amyloid receptor agonists may have an isoelectric point (pi) in the range of 8 to 12, such as 8 to 9. Kagelinide has a theoretically calculated isoelectric point of approximately 8.56. cyclodextrin

The pharmaceutical formulations disclosed herein include cyclodextrins containing hydroxypropyl substituted cyclodextrins. Pharmaceutical formulations may include about 10% to 25% w/v cyclodextrins containing hydroxypropyl substitutes. Pharmaceutical formulations may include more than 10% w/v of cyclodextrins containing hydroxypropyl substitutes. Pharmaceutical formulations may include less than 22% w/v of hydroxypropyl-substituted cyclodextrins. Pharmaceutical formulations may include about 10% to 20% w/v, about 15% to 25% w/v, about 12% to 18% w/v, about 10% to 17.5% w/v, about 11.25% to 15 %, such as about 15% w/v, includes hydroxypropyl-substituted cyclodextrin.

Cyclodextrin is an oligosaccharide starch derivative composed of 6, 7 or 8 α-(1,4)-glucopyranose (glucose) units arranged in a cyclic arrangement, and is expressed as α, β or γ type respectively. Cyclodextrins have wide applications as pharmaceutical excipients [P. Breen & S. S. Jambhekar, Cyclodextrins in pharmaceutical formulations II: solubilization, binding constant, and complexation efficiency), Drug Discovery Today, Volume 21, February 2, 2016]. Guidance on their use as pharmaceutical excipients is described by the European Medicines Agency [Background review for cyclodextrins used as excipients, 2014, EMA/CHMP/333892/2013 , Committee for Human Medicinal Products (CHMP)], [Cyclodextrins used as excipients (Cyclodextrins used as excipients), 2017, EMA/CHMP/333892/2013, Committee for Human Medicinal Products (CHMP)]. Cyclodextrin types without hydrophilic substitution have poor solubility and are rarely used in parenteral drug products.

In order to improve the solubility of cyclodextrins, the hydroxyl groups of the glucose units of the cyclodextrin can be hydrophilically substituted with different numbers of e.g. hydroxypropyl groups, resulting in different degrees of substitution, which can be described as in each cyclodextrin molecule The average number of hydroxypropyl groups (abbreviated DS) or described as the molar degree of substitution, which corresponds to the average number of hydroxypropyl groups per glucose unit present in the relevant cyclodextrin (abbreviated MS). The hydroxypropyl value for each cyclodextrin can be achieved by multiplying the molar degree of substitution by the number of glucose units included in the relevant cyclodextrin. Differences in the degree of substitution can lead to changes in physicochemical properties, such as surface activity and complexing ability. The hydroxyl group can also be chemically substituted by a sulfobutyl ether group. These primarily hydrophilic modifications have resulted in cyclodextrin derivatives that are highly suitable for parenteral administration [Cyclodextrins used as excipients, 2017, EMA/CHMP/333892/2013, Committee for Human Medicinal Products (CHMP)]. The abbreviation for cyclodextrins containing hydroxypropyl substitution is usually HP-CD, while the abbreviation for cyclodextrins containing sulfobutylether substitution is SBE-CD.

Cyclodextrins that include hydrophilic substitutions adopt what can be described as a tapered shape with a hydrophobic inner cavity and a hydrophilic outer surface formed by a number of hydrophilic substituents capable of forming hydrogen bonds with adjacent water molecules, thereby improving water solubility [ T. Loftsson, Cyclodextrins in Parenteral Formulations, Journal of Pharmaceutical Sciences, 2020, 1-11].

Their hydrophobic microenvironment within the cavities of these conical structures allows them to form drug-cyclodextrin complexes primarily through hydrophobic interactions [T. Loftsson, Cyclodextrins in Parenteral Formulations Formulations), Journal of Pharmaceutical Sciences, 2020, 1-11]. When a complex is formed between a cyclodextrin and a drug molecule with one or more hydrophobic regions, these regions, as well as the hydrophobic regions of the cyclodextrin, become obscured by water, thereby increasing the solubility of the complex compared to the solubility of the individual components. . Furthermore, once complexes are formed between cyclodextrins and peptide molecules, they impair the intermolecular interactions that often lead to aggregation [T. Loftsson, Cyclodextrins in Parenteral Formulations], Journal of Pharmaceutical Sciences, 2020, 1-11].

Unexpectedly, cyclodextrins with hydroxypropyl substitution were found to be advantageous in stabilizing coformulations of caglitinide and semaglutide with each other compared to the same cyclodextrin type with sulfobutyl ether substitution. .

The cyclodextrin system may be a hydroxypropyl-substituted alpha form, which includes six glucose units arranged in a ring. The α-type hydroxypropyl-substituted cyclodextrin is abbreviated as HP-A-CD. Hydroxypropyl-α-cyclodextrin (CAS: 128446-33-3/99241-24-4) is commercially available with an average molar substitution (MS) of 0.8 and a molar substitution range of 0.5 to 0.9 .

Pharmaceutical formulations disclosed herein may include a cyclodextrin, which is a hydroxypropyl-substituted beta form, including seven cyclically arranged glucose units.

The β-type hydroxypropyl-substituted cyclodextrin is abbreviated as HP-B-CD.

Hydroxypropyl-β-cyclodextrin is a well-known pharmaceutical excipient that is commonly used in small molecule pharmaceutical formulations, mainly to increase solubility and bioavailability [T. Loftsson, Cyclodextrins in parenteral formulations Cyclodextrins in Parenteral Formulations, Journal of Pharmaceutical Sciences, 2020, 1-11]. To date, the use of cyclodextrins and substituted cyclodextrin derivatives in protein and peptide-based pharmaceutical formulations has been limited.

According to the European and American Pharmacopoeia [USP 38 NF 33, Pharm Eur 8, estimated according to the method described in USP〈761〉/Pharm. Eur. 2.2.33], the hydroxypropyl group available on the market as a pharmaceutical excipient -The degree of hydroxypropyl substitution (DS) of β-cyclodextrin ranges between 2.8 and 10.5, which corresponds to 0.4 to 1.5 hydroxypropyl groups (MS) per glucose unit. Commercially available cyclodextrins, such as hydroxypropyl-β-cyclodextrin, are often described by their mean molar substitution (MS) over their molar substitution range.

Hydroxypropyl-β-cyclodextrin (CAS: 128446-35-5/94035-02-6) is commercially available as an excipient with an average molar substitution (MS) including : MS: 0.62, the molar substitution range is from 0.58 to 0.68; MS: 0.67, the molar substitution range is from (0.6 to 0.9); MS: 0.68, the molar substitution range is from (0.58 to 0.72); MS: 0.84, molar The substitution range is from (0.8 to 1.0); MS: 0.92, the molar substitution range is from (0.81 to 0.99); MS: 1.08, the molar substitution range is from (0.86 to 1.14); where each value describes the molar substitution range per glucose unit Hydroxypropyl quantity.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having a minimum of about 0.4 hydroxypropyl groups per glucose unit. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having up to about 1.0 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having a substitution range of 0.58 to 1.0 hydroxypropyl groups per glucose unit. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having a mean molar substitution (MS) range of about 0.62 to 0.92 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having an average molar substitution (MS) of about 0.62 to 0.84 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.4 to 0.75 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.75 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having a mean molar substitution (MS) per glucose unit of about 0.62. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.58 to 0.68 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-β-cyclodextrin having a mean molar substitution (MS) per glucose unit of about 0.68. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.58 to 0.72 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin, which has a mean molar substitution (MS) of about 0.67. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.6 to 0.9 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin, which has a mean molar substitution (MS) of about 0.84. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.8 to 1.0 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin, which has a mean molar substitution (MS) of about 0.92. Pharmaceutical formulations disclosed herein may include hydroxypropyl-beta-cyclodextrin having about 0.81 to 0.99 hydroxypropyl groups per glucose unit.

Pharmaceutical formulations disclosed herein may include 10% to 25% w/v (such as about 10% to 20% w/v, such as about 15% to 25% w/v, such as about 12% to 18% w/v, such as about 10% to 17.5% w/v, such as about 11.25% to 15%, such as about 15% w/v) hydroxypropyl-β-cyclodextrin having at least about 0.4 hydroxypropyl per glucose unit and up to about 1.0 hydroxypropyl groups per glucose unit; such as an average of 0.62 to 0.92 hydroxypropyl groups per glucose unit, such as about 0.75 hydroxypropyl groups per glucose unit; such as 0.62 to 0.84 hydroxypropyl groups per glucose unit Propyl; such as about 0.4 to 0.75 hydroxypropyl per glucose unit; such as an average of 0.62 hydroxypropyl per glucose unit, such as about 0.58 to 0.68 hydroxypropyl per glucose unit. Other excipients

Pharmaceutical formulations may include buffers. The use of buffers in pharmaceutical formulations is well known to the skilled person. For convenience, reference is made to Remington: The Science and Practice of Pharmacy , 20th Edition, 2000.

pH can be measured at "room temperature", usually defined as 15 to 25°C or 15 to 20°C. The pH value is preferably measured at about 20°C.

Pharmaceutical formulations disclosed herein may include buffers having a pKa close to the desired pH of the solution. Pharmaceutical formulations can include a buffer having a pKa of at least one of about 5.0 to 7.0. Pharmaceutical formulations may include buffers having a pKa of about 5.0 to 7.0. The pharmaceutical formulation may include a buffer selected from the group consisting of histidine, citrate, and/or phosphate. The buffer can be citrate with a concentration of 3-30 mM. The buffer may be histidine, with a concentration of 3-30 mM. The buffer can be phosphate with a concentration of 3 to 30 mM.

Pharmaceutical formulations may further include one or more agents for adjusting pH, such as NaOH and/or HCl.

The desired pH of pharmaceutical formulations may be about 5.5 to 6.5. The pH is preferably 5.6 to 6.0. The pH may be about 5.6, such as about 5.7, such as about pH 5.8, such as about 5.9, such as about 6.0.

Pharmaceutical formulations may include tonicity agents. The use of tonicity agents in pharmaceutical compositions is well known to the skilled person. For convenience, reference is made to Remington: The Science and Practice of Pharmacy , 20th Edition, 2000.

The purpose of tonicity agents is to protect living tissue when the formulation is injected into the body. The tonicity agent may be selected from the group consisting of: mannitol, sorbitol or trehalose, or combinations thereof. In some embodiments, the tonicity agent is mannitol. In some embodiments, the tonicity agent is sorbitol. In some embodiments, the tonicity agent is trehalose.

The concentration of tonicity agent is such that the formulation is isotonic. The tonicity agent is mannitol, which can be present at a concentration of 16.5 to 37.5 mg/ml, such as about 20 mg/ml. Wherein the tonicity agent is sorbitol, which can be present at a concentration of about 10 to 40 mg/ml, such as about 16.5 to 37.5 mg/ml; such as about 10 to 30 mg/ml; such as about 16 to 28 mg/ml, such as about 16.5 to 25 mg/ml, such as about 16 to 26 mg/ml; such as about 16 to 24 mg/ml; such as about 26 mg/ml, such as about 24 mg/ml, such as about 22 mg/ml, such as about 20 mg/ml ml, such as about 18 mg/ml, such as about 16 mg/ml, such as about 12 mg/ml. The tonicity agent is trehalose, which can be present at a concentration of 33 to 75 mg/ml, such as about 38 mg/ml.

Pharmaceutical formulations may include surfactants. Surfactants may also be included to increase the physical stability and robustness of the formulation during preparation, storage, and use as a medicament; for example, retaining formulation stability when exposed to air within the container. The use of surfactants in pharmaceutical compositions is well known to the skilled person. For convenience, reference is made to Remington: The Science and Practice of Pharmacy , 20th Edition, 2000.

The surfactant may be selected from the group consisting of polysorbate 20 and/or polysorbate 80. The surfactant may be polysorbate 20. The surfactant may be polysorbate 80.

Pharmaceutical formulations may include 0.01 mg/ml or more of polysorbate 20 and up to 2.0, such as up to 1.5 mg/ml of polysorbate 20, such as about 0.01 to 1.0 mg/ml of polysorbate 20, Such as approximately 0.05 mg/ml polysorbate 20.

Pharmaceutical formulations may include 0.01 mg/ml or more of Polysorbate 80 and up to 2.0, such as up to 1.5 mg/ml of Polysorbate 80, such as about 0.01 to 1.0 mg/ml of Polysorbate 80, Such as approximately 0.05 mg/ml polysorbate 80.

Pharmaceutical formulations include water for injection (WFI). Pharmaceutical formulations may include more than 75% w/w water, such as 80% w/w water, such as about 85% w/w water, such as up to 90% w/w water.

The pharmaceutical formulations disclosed herein may exclude preservatives. medical use

The pharmaceutical formulations disclosed herein are useful for medical purposes.

The pharmaceutical formulations disclosed herein may be administered by parenteral injection. The pharmaceutical formulations disclosed herein may be administered by subcutaneous injection.

The term "treatment" as used herein refers to the medical treatment of any human or other vertebrate individual in need thereof. The individual should undergo a physical examination by a licensed physician or veterinarian who has made a preliminary or definite diagnosis indicating that the use of the specific treatment would be beneficial to the health of the human or other vertebrate animal. The timing and purpose of treatment may vary from person to person based on the individual's health status. Accordingly, the treatment may be prophylactic (preventable), palliative, symptomatic and/or curative.

In some embodiments, pharmaceutical formulations are administered to human subjects.

The pharmaceutical formulations disclosed herein can be used for: (i) Prevention and/or treatment of all forms of diabetes and related symptoms, such as hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin-dependent diabetes, MODY (adult-onset diabetes of the young) , gestational diabetes, and/or reduced HbA1c; (ii) Delay or prevent the progression of diabetic disease, such as type 2 diabetes, delay the progression of impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, and/or delay the progression of insulin-requiring type 2 diabetes. Type 2 diabetes requiring insulin; (iii) Prevent and/or treat eating disorders, such as obesity, for example, by reducing food intake, reducing body weight, suppressing appetite, inducing satiety; treating and/or preventing bulimia, eating impulses, bulimia nervosa and/or or obesity induced by administration of antipsychotics or steroids; reduced gastrointestinal motility; and/or delayed gastric emptying; (iv) Prevent and/or treat cardiovascular disease, such as delaying or reducing the development of major adverse cardiovascular events (MACE) selected from: cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, vascular vascular disease plastic, patients hospitalized with unstable angina, and patients hospitalized with heart failure; (v) Prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) and/or non-alcoholic steatohepatitis (NASH); (vi) Prevention and/or treatment of cognitive disorders such as Alzheimer's disease.

In some embodiments, the indication is (i). In some embodiments, the indication is (ii). In a further specific aspect, the indication is (iii). In a further specific aspect, the indication is (iv). In a further specific aspect, the indication is (v). In a further specific aspect, the indication is (vi). In some embodiments, the indication is type 2 diabetes and/or obesity.

Typically, all individuals who suffer from obesity are also considered to be overweight. Disclosed herein is a method of treating or preventing obesity. Disclosed herein is the use of the formulations disclosed herein for treating or preventing obesity. In some embodiments, the individual suffering from obesity is a human, such as an adult or a pediatric (including infants, children, and adolescents).

Body mass index (BMI) is a measurement of body fat based on height and weight. The calculation formula is BMI = weight (kg)/height (meters ² ). A human individual suffering from obesity may have a BMI of 30 kg/ ^m2 or higher; the individual may also be referred to as obese. In some embodiments, the human subject with obesity may have a BMI >35 or a BMI range >30 to <40. In some embodiments, obesity is severe obesity or morbid obesity, wherein the human subject may have a BMI > 40.

Disclosed herein is a method of treating or preventing overweight, optionally in the presence of at least one weight-related comorbidity. Disclosed herein is the use of the formulations disclosed herein for the treatment or prevention of overweight, optionally in the presence of at least one weight-related comorbidity.

In some embodiments, the overweight individual is a human, such as an adult or a pediatric person (including infants, children, and adolescents). In some embodiments, the overweight human individual may have a BMI of 25 kg/m ^or higher, such as a BMI of ²⁷ kg/m or higher. In some embodiments, the overweight human subject has a BMI in the range of 25 to <30 or in the range of 27 to <30.

Elevated BMI increases an individual's risk of developing any of a number of diseases or comorbidities. Weight-related comorbidities may be one of the above, or a combination thereof. In some embodiments, the weight-related comorbidity is selected from the group consisting of hypertension, diabetes (such as type 2 diabetes), dyslipidemia, high cholesterol, and obstructive sleep apnea.

Disclosed herein is a method for reducing body weight. A human undergoing weight reduction may have a BMI of 25 kg/ ^m or higher, such as a BMI of 27 kg/m ^or higher (overweight) or a BMI of 30 kg/m ^or higher (obese). In some embodiments, a human undergoing weight reduction may have a BMI of 35 kg/m ^or higher or a BMI of 40 kg/m or ^higher . The term "weight reduction" may include treating or preventing obesity and/or being overweight.

In some embodiments, administration of pharmaceutical formulations of semaglutide and caglitinide disclosed herein may be used as an adjunct to a reduced calorie diet and increased physical activity in the treatment of at least one weight-related comorbidity (e.g., high blood pressure). blood pressure, type 2 diabetes, jaundice, or dyslipidemia) and an initial body mass index (BMI) of 30 kg/ ^m2 or higher (obese) or 27 kg/ ^m2 or higher (overweight) Chronic weight management in adult patients.

In some embodiments, administration of a pharmaceutical formulation of semaglutide and caglitide disclosed herein can result in >15% body weight loss, such as >20% body weight loss, such as >25% body weight loss, within 26 weeks of initiation of treatment. Such as >30% body weight loss, such as about 15% to 40% body weight loss, such as about 20% to 35% body weight loss, such as about 25% to 30% body weight loss.

In some embodiments, administration of a pharmaceutical formulation of semaglutide and caglitide disclosed herein can result in >15% body weight loss, such as >20% body weight loss, such as >25% body weight loss, within 26 weeks of initiation of treatment. Such as >30% body weight loss, such as about 15% to 40% body weight loss, such as about 20% to 35% body weight loss, such as about 25% to 30% body weight loss.

In some embodiments, administration of a pharmaceutical formulation of semaglutide and caglitinide disclosed herein results in a higher HbA _1c compared to treatment with semaglutide alone as the active ingredient or caglilintide alone as the active ingredient. Decrease, unit is %-point. dose

The pharmaceutical formulations of the present invention may be administered in single doses at predetermined time intervals.

The single dose pharmaceutical formulations disclosed herein may contain doses of any one of an amyloid receptor agonist, such as caglitide, and a GLP-1 receptor agonist, such as semaglutide.

An effective amount of an amyloid receptor agonist, such as caglitinide, and a GLP-1 receptor agonist, such as semaglutide, can be administered to an individual in need thereof.

In some embodiments, the dosage is administered approximately once weekly. In some embodiments, the interval between two fixed doses can be about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days. In preferred embodiments, a fixed maintenance dose is administered approximately every 7 days (once a week).

In some embodiments, the dosage is administered to an individual suffering from any one or more of the diseases or comorbidities listed above. In some preferred embodiments, the dosage is administered to individuals suffering from obesity (body mass index [BMI] ≥30 kg/ ^m2 ). In some preferred embodiments, the patient is overweight (BMI ≥27 kg/m ² to <30 kg/m ² ) and suffers from at least one weight-related comorbidity (e.g., hypertension, type 2 diabetes, or Individuals with dyslipidemia) are administered this dose.

In some embodiments, once-weekly treatment results in statistically significant, dose-dependent reductions in body weight.

In some preferred embodiments, the dose is administered as an adjunct to diet and physical activity to improve glycemic control in adults with type 2 diabetes.

After initiation of treatment, it may be beneficial to administer increasing doses of an amyloid receptor agonist, such as caglitinide, and a GLP-1 receptor agonist, such as semaglutide, to an individual in need thereof. Once the individual has adapted to this treatment, it may be beneficial to administer a maintenance dose of an amyloid receptor agonist (such as caglitide) as well as a GLP-1 receptor agonist (such as semaglutide) to the individual in need .

In some embodiments, treatment is once weekly with a 16-week dose escalation period.

In some embodiments, treatment is weekly and dose escalation occurs approximately weekly.

In some embodiments, treatment is once weekly, with dose escalation occurring approximately every other week.

In some embodiments, treatment is once weekly, with dose escalation occurring approximately every three weeks.

In some embodiments, treatment is weekly, with dose escalation occurring approximately every four weeks.

The dose of the amyloid receptor agonist administered may be about 0.25 to 16 mg, such as about 0.25 to 9.0 mg, such as about 0.25 to 4.5 mg, such as about 0.25 to 2.4 mg.

The dose of caglitinide administered may be about 0.25-16 mg, such as about 0.25 to 9.0 mg, such as about 0.25 to 4.5 mg, such as about 0.25 to 2.4 mg.

In some embodiments, the dose of caglitinide administered is about 0.25 mg.

In some embodiments, the dose of caglitinide administered is about 0.5 mg.

In some embodiments, the dose of caglitinide administered is about 1.0 mg.

In some embodiments, the dose of caglitinide administered is about 1.5 mg.

In some embodiments, the dose of caglitinide administered is about 1.7 mg.

In some embodiments, the dose of caglitinide administered is about 2.4 mg.

In some embodiments, the dose of caglitinide administered is about 3.4 mg.

In some embodiments, the dose of caglitinide administered is about 3.6 mg.

In some embodiments, the dose of caglitinide administered is about 4.5 mg.

In some embodiments, the dose of caglitinide administered is about 7.2 mg.

In some embodiments, the dose of caglitinide administered is about 8.0 mg.

In some embodiments, the dose of caglitinide administered is about 9.0 mg.

In some embodiments, the dose of caglitinide administered is about 16.0 mg.

The dose of GLP-1 receptor agonist administered may be about 0.25 to 16 mg, such as about 0.25 to 9.0 mg, such as about 0.25 to 4.5 mg, such as about 0.25 to 2.4 mg.

The dose of semaglutide administered may be about 0.25 to 16 mg, such as about 0.25 to 9.0 mg, such as about 0.25 to 4.5 mg, such as about 0.25 to 2.4 mg.

In some embodiments, the dose of semaglutide administered is about 0.25 mg.

In some embodiments, the dose of semaglutide administered is about 0.5 mg.

In some embodiments, the dose of semaglutide administered is about 1.0 mg.

In some embodiments, the dose of semaglutide administered is about 1.5 mg.

In some embodiments, the dose of semaglutide administered is about 1.7 mg.

In some embodiments, the dose of semaglutide administered is about 2.4 mg.

In some embodiments, the dose of semaglutide administered is about 3.6 mg.

In some embodiments, the dose of semaglutide administered is about 4.5 mg.

In some embodiments, the dose of semaglutide administered is about 4.8 mg.

In some embodiments, the dose of semaglutide administered is about 6.0 mg.

In some embodiments, the dose of semaglutide administered is about 6.9 mg.

In some embodiments, the dose of semaglutide administered is about 7.2 mg.

In some embodiments, the dose of semaglutide administered is about 8.0 mg.

In some embodiments, the dose of semaglutide administered is about 9.0 mg.

In some embodiments, the dose of semaglutide administered is about 12 mg.

In some embodiments, the dose of semaglutide administered is about 16.0 mg. In some embodiments, the dose of semaglutide administered is about 16.0 mg.

In some embodiments, the ratio of amyloid receptor agonist to GLP-1 receptor agonist is about 1:2. In some embodiments, the ratio of caglitide to semaglutide is about 1:2.

In some embodiments, the dose of caglitide is about 0.125 mg and the dose of semaglutide is about 0.25 mg.

In some embodiments, the dose of caglitide is about 0.25 mg and the dose of semaglutide is about 0.5 mg.

In some embodiments, the dose of caglitide is about 0.5 mg and the dose of semaglutide is about 1.0 mg.

In some embodiments, the dose of caglitide is about 0.75 mg and the dose of semaglutide is about 1.5 mg.

In some embodiments, the dose of caglitide is about 0.85 mg and the dose of semaglutide is about 1.7 mg.

In some embodiments, the dose of caglitide is about 1.2 mg and the dose of semaglutide is about 2.4 mg.

In some embodiments, the dose of caglitide is about 2.25 mg and the dose of semaglutide is about 4.5 mg.

In some embodiments, the dose of caglitide administered is about 3.6 mg and the dose of semaglutide is about 7.2 mg.

In some embodiments, the dose of caglitide is about 4.0 mg and the dose of semaglutide is about 8.0 mg.

In some embodiments, the dose of caglitide is about 7.2 mg and the dose of semaglutide is about 14.4 mg.

In some embodiments, the dose of caglitide is about 8.0 mg and the dose of semaglutide is about 16.0 mg.

In some embodiments, the maintenance dose of caglitide is about 1.2 mg and the maintenance dose of semaglutide is about 2.4 mg.

In some embodiments, the maintenance dose of caglitide is about 2.25 mg and the dose of semaglutide is about 4.5 mg.

In some embodiments, the maintenance dose of caglitinide is about 4.0 mg and the maintenance dose of semaglutide is about 8.0 mg.

In some embodiments, the maintenance dose of caglitinide is about 8.0 mg and the maintenance dose of semaglutide is about 16.0 mg.

In some embodiments, the ratio of amyloid receptor agonist to GLP-1 receptor agonist is about 1:1. In some embodiments, the ratio of caglitide to semaglutide is about 1:1.

In some embodiments, the dose of caglitide is about 0.25 mg and the dose of semaglutide is about 0.25 mg.

In some embodiments, the dose of caglitide is about 0.5 mg and the dose of semaglutide is about 0.5 mg.

In some embodiments, the dose of caglitide is about 1.0 mg and the dose of semaglutide is about 1.0 mg.

In some embodiments, the dose of caglitide is about 1.7 mg and the dose of semaglutide is about 1.7 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 2.4 mg.

In some embodiments, the maintenance dose of caglitinide is about 2.4 mg and the maintenance dose of semaglutide is about 2.4 mg.

In some embodiments, the dose of caglitide is about 4.5 mg and the dose of semaglutide is about 4.5 mg.

In some embodiments, the dose of caglitide is about 8.0 mg and the dose of semaglutide is about 8.0 mg.

In some embodiments, the dose of caglitide is about 16.0 mg and the dose of semaglutide is about 16.0 mg.

In some embodiments, the ratio of amyloid receptor agonist to GLP-1 receptor agonist is between 1:1 and 1:7.

In some embodiments, the dose of caglitinide is about 2.4 mg and the dose of semaglutide is about 2.4 mg to 16.0 mg.

In some embodiments, the dose of caglitinide is about 2.4 mg and the dose of semaglutide is about 3.6 mg to 16.0 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 2.4 mg to 13.5 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 3.6 mg to 13.5 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 3.6 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 4.8 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 6.0 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 6.9 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 7.2 mg.

In some embodiments, the dosage of caglitide is about 2.4 mg and the dosage of semaglutide is about 8.0 mg.

In some embodiments, the dose of caglitide is about 2.4 mg and the dose of semaglutide is about 12 mg.

In some embodiments, the dose of caglitide is about 3.4 mg and the dose of semaglutide is about 13.5 mg.

In some embodiments, caglitinide and semaglutide are administered once weekly at an initial dose of 0.25 mg and then escalated to subsequent 0.5 mg, 1.0 mg, and 1.7 mg dose levels until a target of 2.4 mg once weekly is achieved/ Maintenance dose.

In some embodiments, caglitide and semaglutide are administered at 0.25 mg once weekly and then escalated to subsequent 0.5 mg, 1.0 mg, and 1.7 mg dose levels every four weeks until a target of 2.4 mg once weekly is achieved/ Maintenance dose.

In some embodiments, caglitide and semaglutide are administered at 0.25 mg once weekly and then escalated to subsequent 0.5 mg, 1.0 mg, and 1.7 mg dose levels every four weeks until a target of 2.4 mg once weekly is achieved/ Maintenance dose.

In some embodiments, 0.25 mg caglitide and 0.25 mg semaglutide are administered once weekly for four weeks (weeks 0 to 3) and then titrated to subsequent 0.5 mg caglitide and 0.5 mg semaglutide every four weeks ( 4 to 7 weeks), 1.0 mg caglitide and 1.0 mg semaglutide (8 to 11 weeks), and 1.7 mg caglitide and 1.7 mg semaglutide (12 to 15 weeks) at dose levels up to once weekly Target/maintenance doses of 2.4 mg caglitide and 2.4 mg semaglutide (16 weeks and beyond).

In some embodiments, caglitide and semaglutide are administered once weekly at an initial dose of 0.25 mg and then escalated to subsequent dose levels of 0.5 mg, 1.0 mg, 1.7 mg, and 2.4 mg until reaching 4.5 mg once weekly target/maintenance dose.

In some embodiments, caglitide and semaglutide are administered once weekly at an initial dose of 0.25 mg and then escalated to subsequent dose levels of 0.5 mg, 1.0 mg, 1.7 mg, 2.4 mg, 3.6 mg, and 4.5 mg until Achieve target/maintenance dose of 7.2 mg once weekly.

In some embodiments, caglitide and semaglutide are administered once weekly at an initial dose of 0.25 mg and then escalated to subsequent doses of 0.5 mg, 1.0 mg, and 1.7 mg, 2.4 mg, 3.6 mg, 4.5 mg, and 7.2 mg levels until reaching the target/maintenance dose of 8.0 mg once weekly.

In some embodiments, caglitide and semaglutide are administered once weekly at an initial dose of 0.25 mg and then titrated to subsequent doses of 0.5 mg, 1.0 mg, and 1.7 mg, 2.4 mg, 3.6 mg, and 4.5 mg, 7.2 mg, and 8.0 mg dose level until reaching a target/maintenance dose of 16.0 mg once weekly.

As used herein, specific values in relation to numbers or intervals may be interpreted as meaning specific values or approximate values (such as when quantities may be provided by weight, plus or minus 10%, 15% or 20%; such as when measuring pH , plus or minus 0.4).

The following is a non-limiting list of embodiments of the invention. Example 1. A liquid pharmaceutical formulation comprising an amyloid receptor agonist, a GLP-1 receptor agonist and a hydroxypropyl-substituted cyclodextrin. 2. The liquid pharmaceutical formulation according to embodiment 1, wherein the GLP-1 receptor agonist has an isoelectric point that is incompatible with the optimal pH of the amylin receptor agonist. 3. The liquid pharmaceutical formulation according to any one of the preceding embodiments, wherein the optimal pH of the GLP-1 receptor agonist and the amylin receptor agonist differ by at least about two pH units, such as 2 to 5 pH units, such as 2 to 4 pH units, such as 3 to 5 pH units. 4. The liquid pharmaceutical formulation according to any one of the preceding embodiments, wherein the optimal pH of the amyloid receptor agonist is 3.5 to 4.5, such as about 4.0. 5. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the amyloid receptor agonist is caglitinide. 6. The liquid pharmaceutical formulation according to any one of the preceding embodiments, wherein the GLP-1 receptor agonist has an isoelectric point of less than 5.0, such as 3.0 to 5.0, such as 3.8 to 4.9. 7. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the GLP-1 receptor agonist is semaglutide. 8. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the cyclodextrin is a hydroxypropyl-substituted alpha form comprising six cyclically arranged glucose units, or a hydroxypropyl-substituted alpha form comprising seven cyclically arranged glucose units. Hydroxypropyl substituted beta form. 9. A pharmaceutical formulation according to any one of the preceding embodiments, wherein the cyclodextrin is a hydroxypropyl-substituted alpha form comprising six annularly arranged glucose units. 10. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the cyclodextrin is a hydroxypropyl-substituted beta form comprising seven annularly arranged glucose units. 11. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes up to about 1.0 hydroxypropyl groups per glucose unit. 12. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes up to about 0.92 hydroxypropyl groups per glucose unit. 13. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes up to about 0.75 hydroxypropyl groups per glucose unit. 14. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes up to about 0.68 hydroxypropyl groups per glucose unit. 15. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes at least about 0.4 hydroxypropyl groups per glucose unit. 16. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes at least about 0.58 hydroxypropyl groups per glucose unit. 17. The pharmaceutical formulation according to any one of the preceding embodiments, wherein the hydroxypropyl-β-cyclodextrin includes about 0.58 to 1.0 hydroxypropyl groups per glucose unit. 18. The pharmaceutical formulation according to embodiment 17, wherein the hydroxypropyl-β-cyclodextrin includes an average (MS) of 0.62 to 0.92 hydroxypropyl groups per glucose unit. 19. The pharmaceutical formulation according to embodiment 17, wherein the hydroxypropyl-β-cyclodextrin includes an average (MS) of about 0.62 to 0.84 hydroxypropyl groups per glucose unit. 20. The pharmaceutical formulation of embodiment 17, wherein the hydroxypropyl-beta-cyclodextrin includes an average (MS) of about 0.62 hydroxypropyl groups per glucose unit. 21. The pharmaceutical formulation according to embodiment 17, wherein the hydroxypropyl-β-cyclodextrin includes about 0.4 to 0.75 hydroxypropyl groups per glucose unit, such as about 0.58 to 0.68 hydroxypropyl groups per glucose unit . 22. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 10% to 25% w/v cyclodextrin. 23. A pharmaceutical formulation according to any one of the preceding embodiments, comprising more than 10% cyclodextrin. 24. A pharmaceutical formulation according to any one of the preceding embodiments, comprising less than 22% w/v cyclodextrin. 25. A pharmaceutical formulation according to any of the preceding embodiments, comprising less than 20% w/v cyclodextrin. 26. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 10% to 20% w/v of said cyclodextrin. 27. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 10% to 17.5% w/v cyclodextrin. 28. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 12% to 18% w/v cyclodextrin. 29. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 11.25% to 15% w/v HP-β-CD. 30. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 15% w/v cyclodextrin. 31. A pharmaceutical formulation according to any one of the preceding embodiments, comprising at least about 1 mg/ml of the GLP-1 receptor agonist. 32. A pharmaceutical formulation according to any one of the preceding embodiments, comprising up to about 22 mg/ml of the GLP-1 receptor agonist. 33. The pharmaceutical formulation according to any one of the preceding embodiments, comprising about 1 to 12 mg/ml of a GLP-1 receptor agonist. 34. A pharmaceutical formulation according to any one of the preceding embodiments, comprising at least about 1 mg/ml of the amyloid receptor agonist. 35. A pharmaceutical formulation according to any one of the preceding embodiments, comprising up to about 22 mg/ml of an amyloid receptor agonist. 36. The pharmaceutical formulation according to any one of the preceding embodiments, comprising about 1 to 12 mg/ml of an amyloid receptor agonist. 37. A pharmaceutical formulation according to any one of the preceding embodiments, comprising 0.25 to 22 mg/ml of caglitinide. 38. A pharmaceutical formulation according to any one of the preceding embodiments, comprising 0.25 to 22 mg/ml semaglutide. 39. A pharmaceutical formulation according to any one of the preceding embodiments, comprising 0.25 to 22 mg/ml of caglitide and 0.25 to 22 mg/ml of semaglutide. 40. A pharmaceutical formulation according to any one of the preceding embodiments, comprising an effective amount of caglitide and semaglutide. 41. A pharmaceutical formulation according to any one of the preceding embodiments, further comprising a tonicity agent; provided that the tonicity agent is not sodium chloride. 42. The pharmaceutical formulation according to the preceding embodiment, wherein the tonicity agent is mannitol, sorbitol or trehalose, or a combination thereof. 43. The pharmaceutical formulation according to the preceding embodiment, wherein the tonicity agent is mannitol. 44. A pharmaceutical formulation according to the preceding embodiment, comprising mannitol in a concentration of about 16.5 to 37.5 mg/ml, such as about 20 mg/ml. 45. The pharmaceutical formulation according to embodiment 41, wherein the tonicity agent is sorbitol. 46. A pharmaceutical formulation according to the preceding embodiment, comprising a concentration of about 10 to 40 mg/ml (such as about 10 to 30 mg/ml, such as about 16 to 28 mg/ml, such as about 16.5 to 37.5 mg/ml, such as About 16.5 to 25 mg/ml, such as about 16 to 24 mg/ml; such as about 24 mg/ml, such as about 20 mg/ml, such as about 16 mg/ml, such as about 12 mg/ml) sorbitol. 47. The pharmaceutical formulation according to embodiment 41, wherein the tonicity agent is trehalose. 48. A pharmaceutical formulation according to the preceding embodiment, comprising trehalose in a concentration of about 33 to 75 mg/ml, such as about 33 to 45 mg/ml, such as about 38 mg/ml. 49. The pharmaceutical formulation according to any one of the preceding embodiments, further comprising a buffer having a pKa of about 5.0 to 7.0. 50. The pharmaceutical formulation according to any one of the preceding embodiments, further comprising a buffer selected from the group consisting of: histidine, citrate and/or phosphate. 51. A pharmaceutical formulation according to any one of the preceding embodiments, comprising up to 30 mM buffer. 52. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 3 to 30 mM citrate. 53. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 3 to 30 mM histidine, such as 3 to 15 mM histidine, such as 3 to 10 mM histidine, such as about 6 mM Histamine. 54. A pharmaceutical formulation according to any one of the preceding embodiments, comprising about 3 to 30 mM phosphate. 55. The pharmaceutical formulation according to any one of the preceding embodiments, further comprising a surfactant. 56. The pharmaceutical formulation according to the previous embodiment, wherein the surfactant is polysorbate 20 and/or polysorbate 80. 57. The pharmaceutical formulation according to the preceding embodiment, comprising up to about 2.0 mg/ml of polysorbate 20 and/or polysorbate 80. 58. The pharmaceutical formulation according to the preceding embodiment, comprising up to about 1.5 mg/ml of polysorbate 20 and/or polysorbate 80. 59. The pharmaceutical formulation according to the preceding embodiment, wherein the surfactant is polysorbate 80. 60. Pharmaceutical formulation according to any one of the preceding embodiments, wherein the pH is from about 5.5 to 6.5, preferably from 5.6 to 6.0, such as about 5.7, such as about pH 5.8, such as about 5.9, such as about 6.0. 61. A pharmaceutical formulation according to any of the preceding embodiments, comprising at least 75% w/w water, such as about 80% w/w water, such as about 85% w/w water, such as up to about 90% w/w water /w of water. 62. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitide and semaglutide, hydroxypropyl beta-cyclodextrin, including at least about 0.4 per glucose unit Hydroxypropyl and up to about 1.0 hydroxypropyl per glucose unit, histidine, sorbitol, polysorbate 80, and about 75 to 90% w/w water; and having a pH of 5.6 to 6.0. 63. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitinide and semaglutide, hydroxypropyl beta-cyclodextrin, including up to about 0.58 to 100% per glucose unit. 1.0 hydroxypropyl groups, histidine, sorbitol, polysorbate 80, and about 75 to 90% w/w water; and having a pH of 5.6 to 6.0. 64. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitide and semaglutide, hydroxypropyl beta-cyclodextrin, including an average of 0.62 to 0.92 per glucose unit hydroxypropyl, histidine, sorbitol, polysorbate 80, and about 75 to 90% w/w water; and has a pH of 5.6 to 6.0. 65. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitinide and semaglutide, hydroxypropyl beta-cyclodextrin, including an average of 0.62 to 0.84 per glucose unit hydroxypropyl, histidine and/or citrate, sorbitol, polysorbate 80 and about 75 to 90% w/w water; and having a pH of 5.6 to 6.0. 66. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitide and semaglutide, hydroxypropyl beta-cyclodextrin, including an average of 0.62 to 0.68 per glucose unit hydroxypropyl, histidine and/or citrate, sorbitol, polysorbate 80 and about 75 to 90% w/w water; and having a pH of 5.6 to 6.0. 67. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitide and semaglutide, hydroxypropyl beta-cyclodextrin, including an average of 0.62 hydroxyl groups per glucose unit. Propyl, histidine and/or citrate, sorbitol, polysorbate 80 and about 75 to 90% w/w water; and having a pH of 5.6 to 6.0. 68. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitinide and semaglutide, hydroxypropyl beta-cyclodextrin, including up to about 0.75 per glucose unit Hydroxypropyl, such as about 0.4 to 0.75 hydroxypropyl per glucose unit, histidine, sorbitol, polysorbate 80, and about 75 to 90% w/w water; and having a pH of 5.5 to 6.5. 69. The pharmaceutical formulation according to any one of the preceding embodiments, consisting essentially of: effective amounts of caglitinide and semaglutide, hydroxypropyl beta-cyclodextrin, including up to about 0.75 per glucose unit Hydroxypropyl, such as about 0.4 to 0.75 hydroxypropyl per glucose unit, histidine and/or citrate, sorbitol, polysorbate 20 and/or 80 and about 75 to 90% w/w water; and has a pH of 5.6 to 6.0. 70. A pharmaceutical formulation according to any of the preceding embodiments, consisting essentially of: - an effective amount of caglitide and semaglutide, - more than 10% w/v and less than 22% w/v, such as 10% to 20% w/v hydroxypropyl-β-cyclodextrin (0.58 to 1.0 hydroxypropyl groups per glucose unit), - approximately 3 to 30 mM histidine, - approximately 10 to 40 mg /ml sorbitol, - up to 2.0 mg/ml polysorbate 20 and/or polysorbate 80, - a pH value of 5.6-6.0, preferably a pH value of 5.8, - water for injection. 71. A pharmaceutical formulation according to any of the preceding embodiments, consisting essentially of: - an effective amount of caglitide and semaglutide, - more than 10% w/v and less than 22% w/v, such as 10% to 20% w/v hydroxypropyl-beta-cyclodextrin, including an average of 0.62 to 0.84 hydroxypropyl groups per glucose unit, - about 3 to 30 mM histidine and/or citric acid Salt, - about 10 to 40 mg/ml sorbitol, - up to 2.0 mg/ml polysorbate 20 and/or polysorbate 80, - a pH value of 5.6-6.0, preferably a pH value of 5.8, - Water for Injection. 72. A pharmaceutical formulation according to any of the preceding embodiments, consisting essentially of: - 0.25 to 22 mg/ml caglitinide, - 0.25 to 22 mg/ml semaglutide, - more than 10% w/v and less than 22% w/v, such as 10% to 20% w/v hydroxypropyl-beta-cyclodextrin (0.58 to 1.0 hydroxypropyl groups per glucose unit), - about 6 mM histamine Acid, - about 10 to 40 mg/ml sorbitol, - up to 2.0 mg/ml polysorbate 20 and/or 80, - a pH value of 5.6-6.0, preferably a pH value of 5.8, - water for injection. 73. A pharmaceutical formulation according to any of the preceding embodiments for use as a medicament. 74. A pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 for use in treating an individual with an initial body mass index (BMI) of 27 or above, such as 30 or above. 75. The pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 for the treatment of an individual with an initial body mass index (BMI) of 27 or higher and with at least one weight-related comorbidity. 76. The pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 for use as an adjunct to a reduced calorie diet and increased physical activity in the presence of at least one weight-related comorbidity. Adult individuals with an initial body mass index (BMI) of 30 kg/ ^m2 or higher (obese) or 27 kg/ ^m2 or higher (overweight) undergo chronic weight management. 77. Use according to any one of embodiments 73 to 76, wherein the comorbidity is diabetes and/or cardiovascular disease and/or NASH. 78. A pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 for use in treating an individual suffering from diabetes, such as Type II diabetes. 79. The pharmaceutical formulation according to any one of the preceding embodiments 1 to 72, for use as an adjunct to diet and exercise to improve glycemic control in adult individuals with type 2 diabetes. 80. The pharmaceutical formulation according to any one of the preceding embodiments 1 to 72, for the treatment and/or prevention of cardiovascular diseases. 81. The pharmaceutical formulation according to any one of the preceding embodiments 1 to 72, for the treatment and/or prevention of NASH. 82. The pharmaceutical formulation according to any one of the preceding embodiments 1 to 72, for the treatment and/or prevention of cognitive impairment, such as diseases caused by Alzheimer's disease. 83. Pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 according to any one of embodiments 73 to 82, characterized in that the formulation is administered by parenteral injection. 84. Pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 according to any one of embodiments 73 to 82, characterized in that the formulation is administered by subcutaneous injection. 85. Pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 according to any one of embodiments 73 to 84, characterized in that the formulation is administered approximately once a week. 86. A pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 according to any one of embodiments 73 to 85, wherein the dose of caglitinide administered and the dose of semaglutide administered The ratio is approximately 1:1. 87. A pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 according to any one of embodiments 73 to 85, wherein a dose of caglitinide is administered and a dose of semaglutide is administered. The ratio is from 1:1 to 1:7. 88. A pharmaceutical formulation according to any one of the preceding embodiments 1 to 72 according to any one of embodiments 73 to 85, wherein a dose of caglitinide is administered and a dose of semaglutide is administered. The ratio is about 1:2. Examples Example 1 : Effect of hydroxypropyl -β- cyclodextrin (HP-β-CD) on the chemical stability of caglitinide

This example demonstrates the impact of HP-B-CD on the chemical stability of caglitinide in terms of degradation products through the purity of caglitinide and the high molecular weight protein (HMWP) parameters related to caglitinide. Caglitinide is most stable at pH 4.0, and its rate of chemical degradation generally accelerates with increasing pH. Unexpectedly, this example shows that when formulated with HP-B-CD, stable caglitinide formulations are obtained at pH 6. Composition

The compositions of caglitinide formulations 1, 2, and 3 are shown in Table 1. Table 1 Composition of caglitinide formulations 1, 2 and 3 Element caglitinide formulation 1 2 3 Caglitide drug substance (mg/ml) 18 18 18 HP-B-CD KLEPTOSE ^® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ 0 0 25 Citrate, 1 H ₂ O ² (mM) 6.1 3.7 3.7 Disodium hydrogen phosphate, 2 H ₂ O ² (mM) 7.7 12.6 12.6 HCl qs qs qs NaOH qs qs qs Water for injection (WFI) Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 4.0 6.0 6.0 ¹ MS: Molar substitution, corresponding to hydroxypropyl per glucose unit ² Use different buffer concentrations in different formulations to ensure buffering processes at different pHs

Formulations are prepared by dissolving the excipients in water, and dissolving the drug substance in the excipient solution. Adjust solution pH and add water to reach final desired volume before sterilizing via filtration through a 0.22 µm sterile filter. After filtration is complete, fill the formulation into a 1 ml prefilled syringe. method

Samples were stored at 37°C for up to 21 days and analyzed after 14 and 21 days to determine the degree of purity of HMWP and caglitinide.

The extent of covalently bound HMWP was quantified using size exclusion chromatography (SEC). Samples were analyzed using a WATERS HMWP column (7.8 x 300 mm) consisting of 500 mM sodium chloride, 10 mM sodium phosphate dibasic monohydrate, 5 mM orthophosphate, and 50% (v/v) isopropyl alcohol. The eluent was used for isocratic elution. Chromatography was performed at 50°C with UV detection (215 nm) using an injection volume of 10 μl and a flow rate of 0.5 ml/min. HMWP is quantified as the area of all components eluting before the main peak divided by the area of the main peak x 100%.

The purity of caglitinide was determined using reverse phase high performance liquid chromatography (RP-UHPLC). Samples were analyzed using a Kinetex C18, 1.7 μm, 100 Å column (2.1 x 150 mm) with eluent A consisting of 90% v/v 0.09 M phosphate, pH 3.6, and 10% v/v acetonitrile and Gradient elution was performed with eluent B consisting of 60% v/v acetonitrile and 20% v/v isopropanol. Chromatography was performed at 60°C with UV detection (215 nm) using an injection volume of 2 to 7.5 μl and a flow rate of 0.25 ml/min. Purity is estimated as the area of the main peak divided by the area of all peaks x 100%. Table 2 Chemical purity (%) of caglitinide with or without HP-B-CD at pH 4.0 and 6.0 caglitinide formulation pH HP-B-CD (MS: 0.92) ¹ Relationship between HMWP content (%) and time (days) at 37°C Purity (%) of caglitinide versus time (days) at 37°C 0 14 twenty one 0 14 twenty one 1 4.0 0%w/v 0.0% 0.2% 0.2% 90.4% 88.2% 87.4% 2 6.0 0%w/v 0.1% 7.7% 9.8% 90.1% 74.0% 67.1% 3 6.0 25%w/v 0.1% 0.3% 0.5% 90.2% 79.9% 75.3% ¹ MS: molar substitution, summary of hydroxypropyl groups corresponding to each glucose unit

Table 2 shows that when caglitinide was stored at 37°C and at pH 4.0, very low HMWP was formed and only a slight decrease in the purity of caglitinide was seen. In contrast, when the pH was 6.0, the rate of HMWP formation and the decrease in purity of caglitinide accelerated. Unexpectedly, this rapid chemical degradation was counteracted by the addition of HP-B-CD to the formulation, allowing the formulation of caglitinide at pH 6. Example 2 : Effect of HP-B-CD on the physical stability of semaglutide

This example shows the stabilizing effect of HP-B-CD on the physical stability of semaglutide, i.e. its tendency to form peptide fibers. This effect is evident when semaglutide is formulated at suboptimal pH. Composition

The compositions of semaglutide formulations 1, 2, and 3 are shown in Table 3. Table 3 Composition of caglitinide formulations 1, 2 and 3 Element Semaglutide formulations 1 2 3 Semaglutide drug substance (mg/ml) 4.8 4.8 4.8 HP-B-CD KLEPTOSE ^® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ 0 25 0 Citrate, 1 H ₂ O ² (mM) 3.7 3.7 0.9 Disodium hydrogen phosphate, 2 H ₂ O ² (mM) 12.6 12.6 18.2 HCl qs qs qs NaOH qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 6.0 6.0 7.4 ¹ MS: Molar substitution, corresponding to hydroxypropyl per glucose unit ² Use different buffer concentrations in different formulations to ensure buffering processes at different pHs

The formulations were prepared as described in Example 1. method

Semaglutide's tendency to aggregate and form peptide fibrils was measured using thioflavin T (ThT) fluorescence stress assay. The analysis of the presence of peptide fibers is based on the fluorescence properties of the ThT probe, which shows low fluorescence in the unbound/native peptide-bound state, but shows high fluorescence when bound to the peptide fibers, and the wavelength of maximum fluorescence when bound to the fibers Redshift occurs.

The two samples were pooled and 1400 μl of the sample was added to 28 μl of 1 mM ThT stock solution, and 200 μl of this was transferred to 6 different wells of a 96-well microplate with a glass bead in it. The assay was run on a BMG CLARIOstar fluorescent disk reader equipped with a monochromator at 40°C for 169 hours with dual-track shaking and a speed of 300 rpm, using 450 nm and 480 nm for excitation and emission, respectively. Delay time was measured from the start of the experiment until the onset of fibrosis, shown as increased ThT fluorescence. Table 4 Physical stability of semaglutide at pH 6.0 and 7.4 Semaglutide formulations pH HP-B-CD (MS: 0.92) ³ delay time until fibrosis 1 6.0 0%w/v 2.35 hours ¹ 2 6.0 25%w/v >169 ^hours1,2 3 7.4 0%w/v >169 ^{hours1,2 1} Results are an average of 6 replicates ² No fibrosis was observed in any of the 6 replicates over the 169 hour experimental period ³ MS: Molar substitution, summary of hydroxypropyl groups corresponding to each glucose unit

Semaglutide formulations were subjected to shear stress-inducing conditions and measured by their tendency to form peptide fibers. Unexpectedly, the presence of HP-B-CD was found to inhibit the formation of semaglutide peptide fibrils. When semaglutide was formulated at pH 6 and in the absence of HP-B-CD (semaglutide Formulation 1), fibrosis occurred after 2.35 hours; that is, semaglutide was physically unavailable under these conditions. Stable. When semaglutide was formulated at pH 6 and in the presence of HP-B-CD (semaglutide Formulation 2), no fibrosis was observed throughout the experiment; that is, semaglutide was physically stable. Furthermore, when formulated at pH 6 and in the presence of HP-B-CD (semaglutide Formulation 2), the physical stability of semaglutide was found to be similar to that in the absence of HP-B-CD but under its optimal formulation conditions. The physical stability of semaglutide (Semaglutide Formulation 3) was comparable at pH 7.4. Example 3 : Effect of HP-B-CD on the chemical stability of semaglutide

This example demonstrates the impact of HP-B-CD on the chemical stability of semaglutide in terms of degradation products through semaglutide purity and semaglutide-related high molecular weight protein (HMWP) parameters. Composition

The same formulation as in Example 2 was used. process

The formulations were prepared as described in Example 1. method

HMWP content and semaglutide purity were determined after storage at 37°C on day 0, day 14, and day 21.

The purity of semaglutide was determined using reversed-phase high performance liquid chromatography (RP-HPLC), in which samples were analyzed using a Kinetex C18, 2.6 μm column (4.6 x 150 mm) and dissolved in 90% v/v 0.09 M phosphate Gradient elution was performed using the solution, eluent A consisting of pH 3.6 and 10% v/v acetonitrile, and eluent B consisting of 60% v/v acetonitrile and 20% v/v isopropanol. Chromatography was performed at 30°C with UV detection (210 nm) using an injection volume of 10 to 100 μl and a flow rate of 0.7 ml/min. Purity is quantified as the area of the main peak divided by the area of all peaks x 100%.

The extent of covalently bound HMWP was determined using size exclusion chromatography (SEC). Samples were analyzed using a Waters SEC 1.7 μm column (4.6 x 150 mm) in a solution containing 300 mM sodium chloride, 10 mM sodium phosphate dibasic monohydrate, 5 mM orthophosphate, and 50% v/v 2- Isocratic elution was performed with a plasma eluent consisting of propanol. Chromatography was performed at 50°C with UV detection (280 nm) using an injection volume of 1 to 10 μl and a flow rate of 0.3 ml/min. HMWP is quantified as the area of all components eluting before the main peak divided by the area of the main peak x 100%. Table 5 Semaglutide purity at suboptimal and optimal pH Semaglutide formulations pH HP-B-CD (MS: 0.92) ¹ Semaglutide purity (%) versus time (days) at 37°C Relationship between HMWP content (%) and time (days) at 37°C 0 14 twenty one 0 14 twenty one 1 6.0 0%w/v 96.89% 94.58% 93.48% 0.12% 0.25% 0.35% 2 6.0 25%w/v 96.51% 94.53% 93.73% 0.04% 0.11% 0.11% 3 7.4 0%w/v 96.82% 95.56% 94.81% 0.12% 0.32% 0.40% ¹ MS: molar substitution, summary of hydroxypropyl groups corresponding to each glucose unit

Table 5 shows how semaglutide chemical purity decreases over time. When semaglutide (Semaglutide Formulation 1) was formulated at a pH of 6.0, the chemical purity decreased more rapidly than when it was formulated at an optimal pH of 7.4 (Semaglutide Formulation 3). Unexpectedly, HP-B-CD improved the chemical stability of semaglutide (in terms of purity loss and HMWP formation) when semaglutide (Semaglutide Formulation 2) was formulated at pH 6.0. Example 4 : Effect of molar substitution of HP-B-CD on the physical stability of the co-formulation of caglitide and semaglutide

This example shows the effect of HP-B-CD molar substitution on the physical stability of caglitide and semaglutide. Composition

The compositions of Co-formulation 1 and Co-formulation 2 are shown in Table 6. Table 6 Compositions of co-formulation 1 and co-formulation 2 Element Co-formulations 1 2 Caglitide drug substance (mg/ml) 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ 25 - HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ - 25 Citrate, 1 H ₂ O (mM) 3 3 HCl qs qs NaOH qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.7 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

All samples are stored under pressure conditions, defined as: - Period: 28 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, samples are inverted 360° to simulate patient use outside the refrigerator. Three days a week with 20 spins and two days a week with 40 spins.

The number of microscopic particles present quantified the physical stability of the caglitide and semaglutide mixture and was obtained by microfluidic imaging (MFI, see Sharma, D.K. et al., AAPS J. (2010) ), 12: 455-464 on the principles of MFI technology). The following procedure was performed for each syringe sample analyzed: Experiments were performed at ambient temperature. Liquid was withdrawn from each syringe by first removing the plunger and then pipetting the liquid into the sample container. Samples were transferred to a 96-well deep well plate, which was inserted into the sample processing unit (Bot1) on a Protein Simple MFI™ 5200 instrument equipped with a standard Protein Simple MFI™ 100 μm flow cell. Samples were analyzed via a standard MFI system setup, meaning that liquid was pipetted into a reservoir connected to the flow cell, illuminated by 10 LED light sources (470 nm), and monitored with a digital camera (via magnification) throughout the experiment. optics) to record the contents of the flow cell as a bright field image. Data collection was completed using Protein Simple MVSS software. The image stream recorded throughout the run is processed by the proven Novo Nordisk proprietary software MFI data validator, from which the number of individual particles (normalized in counts per milliliter of assay fluid) is obtained and presented by size; >5 µm, > 10 µm, and >25 µm, which are the standard size ranges for particles that can only be seen under a microscope. It should be noted that the number of particles >5 μm includes all particles with a diameter larger than 5 μm (>5 μm, >10 μm and >25 μm), and the number of particles >10 μm includes all particles with a diameter larger than 10 μm (>10 μm and >25 μm). >25 μm). Particle size is defined as equivalent circle diameter (ECD).

The presence of amyloid peptide fibers was analyzed using a thioflavin-T (ThT) fluorescence assay. Experiments were conducted at 25°C. Liquid was withdrawn from each syringe by first removing the plunger and then pipetting the liquid into the sample container. Subsequently, 500 µl of sample was mixed with approximately 9 µl of ThT stock solution in a separate sample container to give a final ThT concentration of 20 µM. Samples were incubated in the dark at ambient temperature for 25 minutes. Transfer 200 µl of sample to one well of a 96-well microplate. Samples were measured on a BMG CLARIOstar fluorescent disk reader equipped with a monochromator using 440 nm for excitation and 470 to 550 nm for emission.

Data acquisition was completed using CLARIOstar control software. In this assay, the emission maximum is observed to occur at a wavelength of approximately 485 nm; the analytical results are therefore reported as ThT fluorescence at 485 nm, expressed in relative fluorescence units (RFU). Table 8 Physical stability of HP-B-CD co-formulations with high or medium molar substitution Co-formulations HP-B-CD average molar substitution Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 Particle size＞5μm 1 HP-B-CD (MS: 0.92) ¹ 459 334 1754 22433 2 HP-B-CD (MS: 0.62) ¹ 256 379 715 245 Particle size＞10μm 1 HP-B-CD (MS: 0.92) ¹ 52 27 602 10598 2 HP-B-CD (MS: 0.62) ¹ 39 39 132 122 Particle size＞25μm 1 HP-B-CD (MS: 0.92) ¹ 0 0 2 644 2 HP-B-CD (MS: 0.62) ¹ 6 0 8 4 ThT fluorescence at 485 nm 1 HP-B-CD (MS: 0.92) ¹ 3357 3538 3588 13640 2 HP-B-CD (MS: 0.62) ¹ 3784 3720 3967 3463 The result for the number of particles visible under the microscope is the average of 3 replicates and has been rounded to the nearest integer value ¹ MS: molar substitution, corresponding to the sum of hydroxypropyl groups per glucose unit

The results in Table 7 show that the least particles were produced in co-formulation 2 containing HP-B-CD (average MS: 0.62). Furthermore, no microscopic increase in the number of particles or ThT fluorescence was seen in the case of co-formulation 2 during the 28 day experiment.

In contrast, in the case of co-formulation 1 containing HP-B-CD (average MS: 0.92), an increase in the number of particles visible only microscopically after 21 days and after 28 days was seen ThT fluorescence increases.

Among these other identical, citrate-buffered semaglutide and caglitinide co-formulations, co-formulation (2) including HP-B-CD (average MS: 0.62) was better than co-formulations including HP-B Co-formulation (1) of -CD (average MS: 0.92) is physically more stable. Example 5 : Effect of hydroxypropyl -B- cyclodextrin concentration on the chemical stability of semaglutide

This example shows the concentration-dependent effect of HP-B-CD on the chemical stability of semaglutide. Composition

The compositions of co-formulation 3, co-formulation 4, and co-formulation 5 are shown in Table 8. Table 8 Compositions of co-formulation 3, co-formulation 4, and co-formulation 5 Element Co-formulations 3 4 5 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 11.25 12.5 15 L-Histidine (mM) 6 6 6 Sorbitol ² (mg/ml) 26 twenty four 20 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 HCl qs qs qs NaOH qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.7 5.7 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl per glucose unit ² Due to the different concentrations of HP-B-CD tested, different sorbitol concentrations were required to obtain isotonic preparations

The formulations were prepared as described in Example 1. method

Samples were stored at 37°C for 28 days and analyzed after 14, 21, and 28 days to determine the chemical purity of semaglutide.

The purity of semaglutide was determined using reversed-phase ultraperformance liquid chromatography (RP-UHPLC), in which samples were analyzed using a Waters Acquity phenyl-hexyl 1.7 μm column (2.1 x 150 mm) and 0.09% in MQ water. Gradient elution was performed with eluent A consisting of TFA, and eluent B consisting of 0.09% TFA in MQ water, 0.09% TFA in MQ water with 80% acetonitrile. Chromatography was performed at 62°C with UV detection (215 nm) using an injection volume of 2 to 14 μl and a flow rate of 0.25 ml/min. Purity was quantified as the area of the main semaglutide peak divided by the areas of all relevant peaks x 100%.

It should be noted that in other experiments, the same method was used to determine caglitinide purity. Table 9 Chemical purity (%) of semaglutide with different HP-B-CD concentrations Co-formulations HP-B-CD (MS: 0.62) ¹ Semaglutide purity (%) versus time (days) at 37°C 0 14 twenty one 28 3 11.25%w/v 96.90% 90.72% 88.65% 87.52% 4 12.5% w/v 96.87% 92.90% 91.25% 89.81% 5 15%w/v 96.83% 93.61% 92.92% 91.67% ¹ MS: molar substitution, summary of hydroxypropyl groups corresponding to each glucose unit

The results in Table 9 show that semaglutide chemical stability and purity also depend on HP-B-CD concentration. Semaglutide remained chemically stable in all co-formulations including 11.25% to 15% w/v HP-B-CD. However, semaglutide chemical stability and purity were greatest when the co-formulation included 15% w/v HP-B-CD. Example 6 : Effect of different tonicity agents on the physical stability of co-formulations

This example shows the stability effect of different tonicity agents on the physical stability of an otherwise identical co-formulation of caglitide and semaglutide. Composition

The compositions of Co-formulation 6 to Co-formulation 12 are shown in Table 10. Table 10 Compositions of co-formulations 6 to 12 Element Co-formulations 6 7 8 9 10 11 12 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 10 10 10 10 10 10 10 L-Histidine (mM) 6 6 6 6 6 6 6 Polysorbate 20 (mg/ml) 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Glycerin (mg/ml) - 16 - - - - - Sorbitol(mg/ml) - - 34 - - - - Mannitol(mg/ml) - - - 34 - - - Trehalose(mg/ml) - - - - 63 - - Sucrose(mg/ml) - - - - - 63 - NaCl (mg/ml) - - - - - - 6.0 HCl qs qs qs qs qs qs qs NaOH qs qs qs qs qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.7 5.7 5.7 5.7 5.7 5.7 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

All samples are stored under pressure conditions, defined as: ․ Period: 18 days․ Temperature: 37°C ± 2°C ․ Pressure Conditions: During storage, samples were inverted 360° to simulate patient use outside the refrigerated room. 100 spins five days a week. The number of particles visible under the microscope was quantified as described in Example 4. Table 11 Effects of different tonicity agents on the physical stability of co-formulations Co-formulations tonicity agent Number of particles visible under a microscope as a function of time ( days ) 0 7 11 14 18 Particle size＞5μm 6 without 256 385 429 952 1296 7 glycerin 292 241 292 466 691 8 Sorbitol 212 341 340 517 345 9 Mannitol 94 438 177 245 360 10 Trehalose 121 155 213 504 472 11 sucrose 59 104 140 539 749 12 NaCl ¹ 36 946 - - - Particle size＞10μm 6 without 4 27 96 216 377 7 glycerin 10 twenty three twenty three 48 106 8 Sorbitol 6 54 42 36 73 9 Mannitol 2 twenty three 12 8 50 10 Trehalose 4 10 40 50 69 11 sucrose 2 8 13 142 253 12 NaCl ¹ 2 403 - - - Particle size＞25μm 6 without 0 4 4 27 75 7 glycerin 0 0 4 6 4 8 Sorbitol 0 0 2 4 0 9 Mannitol 0 0 0 0 0 10 Trehalose 0 0 0 0 4 11 sucrose 0 0 2 15 55 12 NaCl ¹ 0 106 - - - Results are the average of 2 replicates and have been rounded to the nearest integer value (-) No sampling was performed ^1. Regarding co-formulation 12 containing NaCl, due to the rapid increase in the number of particles that can only be seen under the microscope, sampling was slower than for the other formulations. Things stop earlier. Summary

The results in Table 11 show that the number of microscopic particles increased most rapidly in the coformulation including NaCl as the tonicity agent (Coformulation 12). After 7 days, the number of particles significantly exceeded the number of particles determined from the other co-formulations. Therefore, sampling of the NaCl-containing co-formulations was stopped after 7 days to analyze the number of microscopically visible particles.

After 14 days, an increase in the number of microscopic particles was seen in the co-formulation containing glycerol and sucrose, and the two co-formulations were considered to be comparable in terms of physical stability. Particle numbers remained lowest in co-formulations containing mannitol, sorbitol or trehalose. In these co-formulations, little microscopic increase in the number of particles was seen during the 18-year period when the co-formulations were stored under pressure conditions.

Of the co-formulations tested, those that included mannitol, sorbitol, or trehalose as tonicity agents remained the most stable over time. Example 7 : Effect of different surfactants on the physical stability of co-formulations

This example shows the effect of different surfactants on the physical stability of an otherwise identical co-formulation of caglitide and semaglutide. Composition

The compositions of co-formulation 13, co-formulation 14 and co-formulation 15 are shown in Table 12. Table 12 Compositions of co-formulation 13, co-formulation 14 and co-formulation 15 Element Co-formulations 13 14 15 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 10 10 10 L-Histidine (mM) 6 6 6 Polysorbate 20 (mg/ml) 0.05 - - Polysorbate 80 (mg/ml) - 0.05 - Poloxamer 188 (mg/ml) - - 0.5 HCl qs qs qs NaOH qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.7 5.7 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

All samples are stored under pressure conditions, defined as: ․ Period: 17 days․ Temperature: 37°C ± 2°C ․ Pressure Conditions: During storage, samples were inverted 360° to simulate patient use outside the refrigerated room. 100 spins five days a week. The number of particles visible under the microscope was quantified as described in Example 4. Table 13 Effect of different surfactants on the physical stability of co-formulations Co-formulations surfactant Number of particles visible under a microscope as a function of time ( days ) 0 7 10 14 17 Particle size＞5μm 13 Polysorbate 20 464 304 396 772 2803 14 Polysorbate 80 566 292 520 286 ¹ 348 15 Poloxamer 188 1235 1278 2783 940 ¹ 1704 Particle size＞10μm 13 Polysorbate 20 twenty three twenty three 43 143 297 14 Polysorbate 80 19 31 76 57 ¹ 48 15 Poloxamer 188 100 245 301 290 ¹ 523 Particle size＞25μm 13 Polysorbate 20 0 2 6 11 13 14 Polysorbate 80 0 0 0 4 ¹ 0 15 Poloxamer 188 0 13 17 61 ¹ 52 Results are the average of 2 replicates and have been rounded to the nearest integer value of ^1. Only 1 replicate was summarized .

Co-formulation 14 contained the lowest microscopic particle count when stored under pressure for 17 days. A microscopic increase in particles was observed in coformulation 13 containing polysorbate 20 after 14 days, and in coformulation 15 containing poloxamer 188 after 7 days under stress conditions. Forms particles that can only be seen under a microscope. It is clear that the coformulation containing polysorbate 80 is the most stable, and the coformulation containing polysorbate 20 is also acceptably stable. Example 8 : Effect of different buffer substances on the physical stability of co-formulations

This example shows the effect of buffer materials on the physical stability of otherwise identical co-formulations of caglitide and semaglutide. Composition

The compositions of Co-formulation 1 and Co-formulation 16 are shown in Table 14. Table 14 Compositions of co-formulation 1 and co-formulation 16 Element Co-formulations 1 16 Caglitide drug substance (mg/ml) 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ 25 25 Citrate, 1 H ₂ O (mM) 3 - L-Histidine (mM) - 6 HCl qs qs NaOH qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.7 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

All samples were stored under pressure conditions, defined as: ○ Duration: 21 days ○ Temperature: 37°C ± 2°C ○ Pressure conditions: During storage, samples were inverted 360° to simulate patient use outside the refrigerated room. 100 spins five days a week. The number of particles visible under the microscope was quantified as described in Example 4. Table 15 Effect of buffer substances on the physical stability of co-formulations Co-formulations buffer substance Number of particles visible under a microscope as a function of time ( days ) 0 7 11 14 18 twenty one Particle size＞5μm 1 Citrate 459 668 408 409 4717 3115 16 Histidine 386 752 569 571 2495 2392 Particle size＞10μm 1 Citrate 52 117 55 125 2114 1446 16 Histidine 68 80 103 151 969 760 Particle size＞25μm 1 Citrate 0 0 0 16 542 332 16 Histidine 2 0 4 13 162 117 The result is the average of 2 replicates and has been rounded to the nearest integer value

The physical stability of both co-formulations was similar and acceptable up to day 14 of storage under pressure conditions. However, after 18 days, the number of microscopically visible particles was higher in the citrate-buffered co-formulation (co-formulation 1) than in the histamine-buffered co-formulation (co-formulation 16). many. Histidine buffered co-formulation 16 was the most stable. Example 9 : Effect of different buffer concentrations on the chemical stability of co-formulations

This example shows the effect of buffer concentration on the chemical stability of otherwise identical co-formulations. Composition

The compositions of Co-formulation 17 and Co-formulation 18 are shown in Table 16. Table 16 Compositions of co-formulation 17 and co-formulation 18 Element Co-formulations 17 18 Caglitide drug substance (mg/ml) 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 10 10 L-Histidine (mM) 6 20 HCl qs qs NaOH qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.7 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

Samples were stored at 30°C for 21 days and analyzed after 7, 14 and 21 days to determine the chemical purity of caglitinide. The purity of caglitinide was determined as described in Example 5 (for semaglutide). Table 17 Effect of buffer concentration on the chemical stability of caglitinide in co-formulations Co-formulations buffer concentration Cagelin peptide purity (%) versus time (days) at 30°C 0 7 14 twenty one 17 Histidine: 6 mM 89.2% 88.2% 86.5% 84.2% 18 Histidine: 20 mM 89.2% 87.9% 86.0% 83.5% Summary

The results in Table 17 show that both co-formulations are stable. However, the chemical purity of caglitinide was highest in coformulation 17. The purity of caglitinide decreased rapidly over time when the histidine concentration was 20 mM. Example 10 : Effect of different buffer concentrations on the physical stability of co-formulations

This example shows the effect of histidine buffer concentration on the physical stability of co-formulations. Composition

The compositions of the co-formulations tested are shown in Table 16. process

The formulations were prepared as described in Example 1. method

All samples are stored under pressure conditions, defined as: ․ Period: 18 days․ Temperature: 37°C ± 2°C ․ Pressure Conditions: During storage, samples were inverted 360° to simulate patient use outside the refrigerated room. 100 spins five days a week. The number of particles visible under the microscope was quantified as described in Example 4. Table 18 Effect of buffer concentration on physical stability of co-formulations Co-formulations buffer concentration Number of particles visible under a microscope as a function of time ( days ) 0 7 10 14 18 Particle size＞5μm 17 Histidine: 6 mM 239 392 165 189 266 18 Histidine: 20 mM 50 262 197 1029 926 Particle size＞10μm 17 Histidine: 6 mM 4 8 15 17 46 18 Histidine: 20 mM 2 17 17 461 423 Particle size＞25μm 17 Histidine: 6 mM 0 0 2 0 2 18 Histidine: 20 mM 0 0 0 120 104 Results are the average of 2 replicates and have been rounded to the nearest integer value. Summary

The difference in physical stability of co-formulas 17 and 18 becomes most apparent after 14 days. The data in Table 18 show that the number of microscopic particles seen in coformulation 18 (containing 20 mM histidine) is higher than that seen in coformulation 17 (containing 6 mM histidine). The number of particles can only be seen below. That is, the co-formulation including 6 mM histidine was the most physically stable. Example 11 : Effect of HP-B-CD concentration on subcutaneous tolerability after subcutaneous injection

This example shows the concentration-dependent effects of HP-B-CD on subcutaneous tissue after subcutaneous injection. Composition

The compositions of the tested co-formulation vehicles prepared at different HP-B-CD concentrations are shown in Table 19. Table 19 Compositions of co-formulated vehicles containing different concentrations of HP-B-CD Element Co-formulation carrier 1 2 3 4 5 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 10 12.5 15 17.5 20 L-Histidine (mM) 6 6 6 6 6 Sorbitol ² (mg/ml) 28 twenty four 20 16 12 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 0.05 0.05 HCl qs qs qs qs qs NaOH qs qs qs qs qs WFI to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml pH 6.0 6.0 6.0 6.0 6.0 ¹ MS: Molar substitution, the concentration of hydroxypropyl ² sorbitol corresponding to each glucose unit changes with the concentration of HP-B-CD to maintain isotonic conditions.

The formulation was prepared as described in Example 1, except that the addition of active pharmaceutical ingredient was omitted. method

After subcutaneous administration of a formulation containing HP-B-CD, 5 Landrace × Yorkshire × Duroc (LYD) pigs were tested by using a 25 G size needle and a syringe with a 5 mm stopper to assess skin lesions obtained after subcutaneous administration of 600 µl for 6 days (necropsy) to study local (subcutaneous) tolerance. Skin samples measuring 2x2 cm were collected at necropsy, fixed in neutral buffered formalin, trimmed with a multi-knife, embedded in paraffin, cut into 4 µm thin layers, mounted on glass slides and subsequently Hematoxylin-eosin (HE) staining was performed. The degree of subcutaneous tissue necrosis was assessed using light microscopy and scored on a numerical scale, with code 1 reflecting "no necrosis" and code 4 reflecting "moderate necrosis." A total of 5 skin samples were taken for each co-formulation vehicle. However, due to variations in sectioning subcutaneous tissue for successful assessment of necrosis, not all injection sites can be assigned a score: 1, no necrosis 2, minimal necrosis 3, mild necrosis 4, moderate necrosis assessment contains 10% w The extent of subcutaneous necrosis induced by isotonic co-formulated vehicle preparations of /v to 20% w/v HP-B-CD when injected subcutaneously. The results are presented in Table 20. Table 20 Necrosis fraction of subcutaneous tissue necrosis for co-formulation necrosis 6 days after injection of co-formulation vehicle with different percentages of HP-B-CD Co-formulation carrier HP-B-CD concentration Necrosis, subcutaneous Number of successfully scored skin samples out of total 1 10%w/v 2,2,2,2,3 5 out of 5 2 12.5% w/v 3,3 2 out of 5 3 15%w/v 2,2,3 3 out of 5 4 17.5%w/v 2,2,3 3 out of 5 5 20%w/v 2,3,3,4 4 out of 5 Summary

A correlation was observed between increased HP-B-CD concentration in the co-formulated vehicle and injection site necrosis. In one case, a co-formulated vehicle containing 20% w/v HP-B-CD caused moderate subcutaneous necrosis at the injection site. Formulations containing less than 20% w/v HP-B-CD all produced only mild or minimal subcutaneous necrosis at the injection site. All coformulation vehicles containing 10% to 20% w/v HP-B-CD were tolerated to an acceptable extent, with coformulations containing 10% to 17.5% w/v HP-B-CD being preferred carrier. Example 12 : Effect of different tonicity agents on subcutaneous tolerability during subcutaneous injection

This example shows the effect of any of three different tonicity agents (sorbitol, mannitol and trehalose) on topical tolerability in an otherwise identical isotonic co-formulation vehicle. Composition

The composition carriers of the co-formulations tested are shown in Table 21. Table 21 Composition of isotonic co-formulation vehicles prepared with different tonicity agents Element Co-formulation carrier 6 7 8 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 15 15 L-Histidine (mM) 6 6 6 Polysorbate 20 (mg/ml) 0.1 0.1 0.1 Sorbitol(mg/ml) twenty four - - Mannitol(mg/ml) - 25 - Trehalose(mg/ml) - - 46 HCl qs qs qs NaOH qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare a volume of 1 ml pH 6.0 6.0 6.0 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulation was prepared as described in Example 1, except that the addition of active pharmaceutical ingredient was omitted. method

HP-B-containing was studied in 2 live Landrace Local tolerability of CD and isotonic vehicle preparations of three different tonicity agents administered subcutaneously. The preparation was injected using a syringe equipped with a 25 G size needle and a 5 mm stopper. Approximately 24 hours after injection, necropsy was performed, and skin samples measuring 2x2 cm were fixed in neutral buffered formalin and trimmed into 4 µm thin layers using multiple blades, embedded in paraffin, and subsequently stained with HE. Regarding both samples, the severity of subcutaneous tissue necrosis, inflammatory cell infiltration, and hemorrhagic distribution was assessed by a trained toxicological pathologist using light microscopy and scored on a numerical scale, where code 1 reflects "no abnormality" and code 3 Reflecting "mild severity": 1. No abnormality 2. Very little severity 3. Mild severity Table 22. Subcutaneous tissue necrosis and inflammatory cells 24 hours after subcutaneous injection of a co-formulation vehicle containing three types of tonicity agents Infiltration and hemorrhage distribution severity scores Co-formulation carrier tonicity agent Necrosis inflammatory cell infiltration Bleeding distribution 6 Sorbitol 2,1 2,1 1,1 7 Mannitol 2,2 2,3 1,2 8 Trehalose 1,1 2,2 1,2 Summary

The data presented in Table 22 show that overall sorbitol was the tonicity agent that caused the least severe necrosis, inflammatory cell infiltration, and hemorrhage. These observations confirm that sorbitol is the preferred tonicity agent to achieve good and acceptable subcutaneous tolerability of co-formulations containing active pharmaceutical ingredients. Example 13 : Confirmation of the effect of tonicity type in histamine-buffered compositions and citrate-buffered compositions on subcutaneous tolerability during subcutaneous injection

This experiment examines: (1) the effect of tonicity type on the local tolerance profile following subcutaneous injection of an otherwise identical histamine-buffered co-formulation; and (2) subcutaneous injection of a citrate-buffered composition without a tonicity agent The effect of co-formulation of vehicle on local tolerance profile. Composition

The compositions of the co-formulations evaluated are described in Tables 23a and 23b. Table 23a Composition of isotonic histamine-buffered co-formulation vehicle Element Co-formulations 19 20 Caglitide drug substance (mg/ml) 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 15 L-Histidine (mM) 6 6 Trehalose(mg/ml) 38.6 - Sorbitol(mg/ml) - 20 Polysorbate 80 (mg/ml) 0.05 0.05 HCl qs qs NaOH qs qs WFI to 1 ml to 1 ml pH 6.0 6.0 ¹ MS: Molar substitution, corresponding to hydroxypropyl groups per glucose unit Table 23b Composition of Isotonic Citrate Buffered Co-Formulation Vehicle Element Co-formulation carrier twenty one HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ 25 Citrate, 1 H ₂ O (mM) 3 HCl qs NaOH qs WFI to 1 ml pH 5.7 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

Co-formulations 19 and 21 were prepared as described in Example 1. Formulation Vehicle 21 was prepared as described in Example 1, except that the addition of an active pharmaceutical ingredient was omitted. method

The local tolerance of the co-formulations described in Tables 23a and 23b when administered subcutaneously was studied in 8 live mini-pigs by evaluating the subcutaneously administered samples using a syringe equipped with a 25 G size needle and a 5 mm stopper. Size 750 µl Skin lesions obtained after 6 days (necropsy). Skin samples measuring 2x2 cm were collected at necropsy, fixed in neutral buffered formalin, trimmed with a multi-knife, embedded in paraffin, cut into 4 µm thin layers, mounted on glass slides and subsequently Hematoxylin-eosin (HE) staining was performed. Regarding the specimens, the severity of subcutaneous tissue necrosis was assessed by trained toxicological pathologists using light microscopy and scored on a numerical scale, where code 1 reflects "no abnormality" and code 5 reflects "marked severity": 1, None The results for abnormality 2, minimal severity 3, mild severity 4, moderate severity 5, and significant severity necrosis scores are shown in Tables 24a and 24b. Table 24a Severity scores of subcutaneous tissue necrosis 6 days after injection of histamine-buffered co-formulations Co-formulations Buffer tonicity agent Necrosis scores of four skin samples 19 Histidine Trehalose 1,2,3,3 20 Histidine Sorbitol 2,2,2,2 Table 24b Subcutaneous tissue necrosis severity scores 6 days after injection of citrate buffered co-formulated vehicle Co-formulation carrier Buffer tonicity agent Necrosis scores of eight skin samples twenty one Citrate without 3,4,4,4,4,5,5,5 Summary

The results presented in Table 24a show that the type of tonicity agent included in the formulation affects its local tolerability in vivo. Tonicity agents are associated with subcutaneous necrosis observed at the injection site. Subcutaneous injection of co-formulation 19 including trehalose resulted in two mild necrotic events (score 3). Subcutaneous injection of co-formulation 20 including sorbitol resulted in minimal necrosis (score 2), which is a better outcome. These results confirm that the formulation ratio included 15% w/v HP-B-CD (average MS: 0.62) and sorbitol in this otherwise identical co-formulated vehicle scenario. The formulation of HP-B-CD (average MS: 0.62) and trehalose was better.

Results presented in Table 24b show that three significant necrotic events were observed using Co-Formula 21 vehicle including 25% w/v HP-B-CD (average MS: 0.92), citrate, and no tonicity agent. (Score 5), confirms that this particular co-formulation is not suitable for subcutaneous injection. Example 14 : Effect of different types of hydroxypropyl-substituted cyclodextrins on the physical and chemical stability of caglitinide and semaglutide co-formulations

This example shows hydroxypropyl-α-cyclodextrin (HP-A-CD), hydroxypropyl-β-cyclodextrin (HP-B-CD), hydroxypropyl γ-cyclodextrin (HP-G- CD) Effects on microscopically visible particle formation and chemical degradation of caglitinide in otherwise identical co-formulations of caglitinide and semaglutide. Composition

The compositions of co-formulations 22, 23 and 24 are shown in Table 25. Table 25 Compositions of co-formulations 22 , 23 and 24 Element Co-formulations twenty two twenty three twenty four Caglitide drug substance (mg/ml) 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 HP-A-CD (CycloLab) (% w/v) (Average MS: 0.8, MS range: 0.5-0.9) ¹ 15 - - HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ - 15 - HP-G-CD (CycloLab) (% w/v) (average MS: 0.6, MS range: 0.4-0.7) ¹ - - 15 L-Histidine (mM) 6 6 6 Sorbitol(mg/ml) 20 20 20 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 HCl qs qs qs NaOH qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.8 5.8 5.8 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

Samples used to determine the number of particles visible under a microscope are stored under pressure conditions, defined as: - Period: 42 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, samples are inverted 360° to simulate patient use outside the refrigerator. Three days a week with 20 spins and two days a week with 40 spins. The number of particles visible under the microscope was determined as described in Example 4.

Samples used to determine caglitinide purity were stored at 37°C for up to 42 days. The purity of semaglutide was determined using the following reverse-phase high performance liquid chromatography (RP-HPLC) method, in which samples were analyzed using a Kinetex C18, 2.6 μm column (4.6 x 150 mm) and incubated with 90% v/v 0.09 M phosphoric acid Gradient elution was performed with salt solution, eluent A consisting of pH 3.6 and 10% v/v acetonitrile, and eluent B consisting of 60% v/v acetonitrile and 20% v/v isopropanol. Chromatography was performed at 30°C with UV detection (210 nm) using an injection volume of 10 to 100 μl and a flow rate of 0.7 ml/min. The purity of caglitinide was quantified as the area of the main peak divided by the area of all relevant peaks x 100%.

The purity of semaglutide was determined using the same method in other experiments. Table 26 Physical stability of caglitinide and semaglutide co-formulations formulated with different types of hydroxypropylcyclodextrin Co-formulations Types of hydroxypropyl-substituted cyclins Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 35 42 Particle size＞5μm twenty two HP-A-CD 65 196 270 441 523 690 twenty three HP-B-CD 49 122 61 245 324 503 twenty four HP-G-CD ¹ 453537 - - - - - Particle size＞10μm twenty two HP-A-CD 4 7 14 27 47 41 twenty three HP-B-CD 4 8 5 20 20 116 twenty four HP-G-CD ¹ 107013 - - - - - Particle size＞25μm twenty two HP-A-CD 0 0 0 1 3 0 twenty three HP-B-CD 0 1 1 1 4 15 twenty four HP-G-CD ¹ 316 - - - - - Results for the number of particles visible under the microscope are the average of 3 replicates and have been rounded to the nearest integer value (-) No sampling was performed ¹ Regarding co-formulation 24 containing HP-G-CD, due to the number of particles Rapid increase, sampling stops earlier than other formulations. Table 27 Chemical purity (%) of caglintide in coformulations of caglintide and semaglutide formulated with different types of hydroxypropylcyclodextrin Co-formulations Types of Hydroxypropyl Cyclodextrin Cagelin peptide purity (%) versus time (days) at 37°C 0 14 28 42 twenty two HP-A-CD 94.0% 87.2% 79.0% 71.1% twenty three HP-B-CD 96.8% 88.9% 81.9% 75.0% Summary

The results presented in Table 26 show that in co-formulation 24 (HP-G-CD), a high number of microscopic particles were already observed at time point zero, which precludes the use of HP-G-CD to co-formulate caglitide and semaglutide. After preliminary analysis at time point zero, sampling of co-formulation 24 containing HP-G-CD was discontinued for microscopic particle number analysis. Regarding co-formulation 22 (HP-A-CD) and co-formulation 23 (HP-B-CD), little microscopic increase in the number of particles was observed.

The chemical purity results of caglitinide using HP-A-CD or HP-B-CD presented in Table 27 show that co-formulation 23 containing HP-A-CD was The purity of caglitinide decreased slightly more rapidly in Formulation 22.

Based on the results in Table 26, HP-A-CD or HP-B-CD systems can be used in co-formulations of caglitinide and semaglutide. However, based on the results in Table 27, HP-B-CD is preferred over HP-A-CD for caglitinide and semaglutide due to the higher purity of caglitinide when formulated with HP-B-CD of co-formulations. Example 15 : Effect of the molar substitution degree of HP-B-CD on the physical and chemical stability of the co-formulation of caglitide and semaglutide

This example shows the effect of molar substitution of HP-B-CD on the formation of microscopic particles, HMWP content, and semaglutide in an otherwise identical citrate-buffered coformulation of caglitinide and semaglutide. Effect of chemical purity. Composition

The compositions of co-formulations 25 to 32 are shown in Table 28. Table 28 Composition of citrate-buffered caglitinide and semaglutide co-formulations containing HP-B-CD excipients with varying degrees of hydroxypropyl molar substitution Element Co-formulations 25 26 27 28 29 30 31 32 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 - 25 - - - - - HP-B-CD (CycloLab) (% w/v) (average MS: 0.67, MS range: 0.6-0.9) ¹ - - - 25 - - - - HP-B-CD Cavitron ^® (Ashland) (% w/v) (Average MS: 0.68, MS Range: 0.58-0.72) ¹ - - - - 25 - - - HP-B-CD Trappsol ^® (CTD, Inc.) (% w/v) (Average MS: 0.84, MS Range: 0.8-1.0) ¹ - - - - - 25 - - HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ - 15 - - - - 25 - HP-B-CD Cavitron ^® (Ashland) (%w/v) (Average MS: 1.08, MS Range: 0.86-1.14) ¹ - - - - - - - 25 Citrate, 1 H ₂ O (mM) 3 3 3 3 3 3 3 3 HCl qs qs qs qs qs qs qs qs NaOH qs qs qs qs qs qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.8 5.8 5.8 5.8 5.8 5.8 5.8 5.8 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

Samples used to determine the number of particles visible under a microscope are stored under pressure conditions, defined as: - Period: 28 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, samples are inverted 360° to simulate patient use outside the refrigerator. Three days a week with 20 spins and two days a week with 40 spins.

The number of particles visible under the microscope was quantified as described in Example 4.

Samples used to determine semaglutide purity and HMWP content were stored at 37°C for up to 28 days. Semaglutide purity was determined as in Example 14.

The extent of covalently bound HMWP was determined using size exclusion chromatography (SEC). Samples were analyzed using a Waters SEC 1.7 μm column (4.6 x 150 mm) containing 185 mM sodium chloride, 5 mM sodium phosphate monobasic monohydrate, 3 mM orthophosphate, and 47% (v/v) Isocratic elution was performed with a plasma eluent consisting of isopropyl alcohol. Chromatography was performed at 50°C with UV detection (215 nm) using an injection volume of 1 to 8 μl and a flow rate of 0.3 ml/min. HMWP is quantified as the area of all components eluting before the main peak divided by the area of the main peak x 100%. Table 29 Microscopic particle content in citrate-buffered caglitinide and semaglutide co-formulations containing HP-B-CD excipients with varying degrees of hydroxypropyl molar substitution Co-formulations HP-B-CD average molar substitution ¹ Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 Particle size＞5μm 25 0.62 82 342 1232 25324 26 0.92 89 10108 13381 49863 27 0.62 126 550 370 448 28 0.67 59 356 191 887 29 0.68 77 27 629 4323 30 0.84 97 374 381 7031 31 0.92 106 244 761 3161 32 1.08 95 32626 54994 114586 Particle size＞10μm 25 0.62 11 73 349 4799 26 0.92 9 4007 5128 20604 27 0.62 13 175 93 131 28 0.67 13 103 53 262 29 0.68 6 10 131 1542 30 0.84 18 129 135 2224 31 0.92 32 59 271 765 32 1.08 6 13421 29840 59104 Particle size＞25μm 25 0.62 0 3 39 330 26 0.92 1 627 830 3461 27 0.62 0 3 8 13 28 0.67 0 1 6 27 29 0.68 0 1 5 245 30 0.84 1 3 34 296 31 0.92 3 4 46 56 32 1.08 1 825 7895 15815 The results for the number of particles visible under the microscope are the average of 3 replicates and have been rounded to the nearest integer value ¹ MS: molar substitution, corresponding to hydroxypropyl units per glucose unit Table 30 in hydroxypropyl groups containing different hydroxyl groups HMWP content in citrate- buffered caglitinide and semaglutide co-formulations of HP-B-CD excipients with propyl molar substitution Co-formulations HP-B-CD mole replaces ¹ HMWP content (%) versus time (days) at 37° C 0 14 twenty one 28 25 0.62 0.03% 0.08% 0.13% 0.18% 26 0.92 0.05% 0.12% 0.18% 0.22% 27 0.62 0.04% 0.08% 0.14% 0.15% 28 0.67 0.05% 0.11% 0.17% 0.20% 29 0.68 0.04% 0.09% 0.16% 0.20% 30 0.84 0.05% 0.12% 0.17% 0.21% 31 0.92 0.04% 0.13% 0.22% 0.27% 32 1.08 0.11% 0.47% 0.61% 0.76% ¹ MS: molar substitution, corresponding to hydroxypropyl per glucose unit Table 31 Citrate-buffered caglitinide and semaglutide in HP-B-CD excipients containing different degrees of molar substitution of hydroxypropyl Chemical purity of semaglutide in co-formulation (%) Co-formulations HP-B-CD mole replaces ¹ Semaglutide purity (%) versus time ( days ) at 37°C 0 14 twenty one 28 25 0.62 97.0% 95.3% 94.4% 93.8% 26 0.92 97.0% 95.1% 93.9% 93.4% 27 0.62 97.0% 95.2% 94.3% 93.4% 28 0.67 96.9% 95.0% 94.1% 93.5% 29 0.68 97.0% 95.3% 94.4% 93.5% 30 0.84 97.1% 95.4% 94.6% 93.8% 31 0.92 96.9% 94.4% 93.4% 92.3% 32 1.08 96.6% 91.4% 89.1% 87.5% ¹ MS: molar substitution, summary of hydroxypropyl groups corresponding to each glucose unit

The results presented in Tables 29, 30, and 31 show that physical stability, HWMP formation, and chemical purity of semaglutide are dependent on HP- when studied in citrate-buffered caglitinide and semaglutide co-formulations. B-CD mole replacement. After 28 days, the lowest HMWP content and little increase in particle number were found in coformulation 27 containing 25% w/v HP-B-CD (average MS: 0.62). In contrast, in co-formulation 32 containing 25% w/v HP-B-CD (average MS: 1.08), a large increase in the number of microscopically visible particles was observed after only 14 days and at The highest content of HMWP was also observed after 28 days.

All citrate-buffered caglitide and semaglutide co-formulations containing 25% w/v HP-B-CD (average MS: 0.92 or less) were physically and chemically stable, those containing 25% HP-B-CD w/v had an average MS of 0.68 or less and was the most stable.

A microscopic increase in the number of particles was seen in coformulation 26 containing 15% w/v HP-B-CD (average MS: 0.92) after only 14 days, indicating the effectiveness of this particular coformulation. Physical instability.

Co-formulation 25 containing 15% w/v HP-B-CD (average MS: 0.62) showed acceptable chemical and physical stability.

However, histidine-buffered caglitide and semaglutide co-formulations 33 to 37 containing 15% w/v HP-B-CD are preferred due to their superior physical stability. Using histidine as buffer liquid and sorbitol as tonicity agent, the preferred HP-B-CD molar substitution range is expanded to an average of 0.62 to 0.92 (or 0.58 to 1.0 in total). Example 16 : Effect of the molar substitution degree of HP-B-CD on the physical stability of the co-formulation of caglitide and semaglutide

This example shows the effect of molar substitution of HP-B-CD on microscopic particle content in an otherwise identical histidine-buffered co-formulation of caglitinide and semaglutide. Composition

The compositions of co-formulations 33 to 38 are shown in Table 32. Table 32 Composition of histine-buffered caglitinide and semaglutide co-formulations 33 to 38 containing HP-B-CD excipients with varying degrees of hydroxypropyl molar substitution Element Co-formulations 33 34 35 36 37 38 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 - - - - - HP-B-CD (CycloLab) (% w/v) (average MS: 0.67, MS range: 0.6-0.9) ¹ - 15 - - - - HP-B-CD Cavitron ^® (Ashland) (% w/v) (Average MS: 0.68, MS Range: 0.58-0.72) ¹ - - 15 - - - HP-B-CD Trappsol ^® (CTD, Inc.) (% w/v) (Average MS: 0.84, MS Range: 0.8-1.0) ¹ - - - 15 - - HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ - - - - 15 - HP-B-CD Cavitron ^® (Ashland) (%w/v) (Average MS: 1.08, MS Range: 0.86-1.14) ¹ - - - - - 15 L-Histidine (mM) 6 6 6 6 6 6 Sorbitol(mg/ml) 20 20 20 20 20 20 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 0.05 0.05 0.05 HCl qs qs qs qs qs qs NaOH qs qs qs qs qs qs WFI Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml Prepare to a volume of 1 ml pH 5.8 5.8 5.8 5.8 5.8 5.8 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

Samples used to determine the number of particles visible only under the microscope are stored under pressure conditions, defined as: - Period: 28 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, the samples are inverted for 360° °To simulate the use of patients outside the cold storage room. Three days a week with 20 spins and two days a week with 40 spins. The number of particles visible under the microscope was quantified as described in Example 4. Table 33 Microscopically visible particle numbers in histidine-buffered caglitinide and semaglutide co-formulations containing HP-B-CD with varying degrees of hydroxypropyl molar substitution Co-formulations HP-B-CD mole replaces ¹ Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 Particle size＞5μm 33 0.62 19 300 252 460 34 0.67 37 176 220 66 35 0.68 twenty two 175 453 290 36 0.84 17 270 221 112 37 0.92 51 873 804 473 38 1.08 71 1311 1765 1621 Particle size＞10μm 33 0.62 1 14 6 61 34 0.67 6 9 6 4 35 0.68 1 5 twenty three 56 36 0.84 1 13 0 6 37 0.92 3 45 twenty four 117 38 1.08 4 143 416 579 Particle size＞25μm 33 0.62 0 0 0 3 34 0.67 0 0 0 0 35 0.68 0 3 0 1 36 0.84 0 0 0 4 37 0.92 0 1 1 20 38 1.08 1 0 14 27 The result for the number of particles visible under the microscope is the average of 3 replicates and has been rounded to the nearest integer value ¹ MS: molar substitution, corresponding to the sum of hydroxypropyl groups per glucose unit

Results presented in Table 33 show that histidine-buffered co-formulations 33 to 37 including HP-B-CD with extensive molar substitution (average MS: 0.62 to 0.92) remained physically stable for 28 days.

In contrast, co-formulation 38 including HP-B-CD (average MS: 1.08) was not physically stable after 14 days.

The results demonstrate synergy between cyclodextrins and other excipients in the co-formulation, broadening the preferred range of molar substitution (average MS: 0.62 to 0.92). Example 17 : Effect of β- cyclodextrin substitution type on physical stability of co-formulations

This example shows the effect of sulfobutoxide-B-cyclodextrin (SBE-B-CD) and hydroxypropyl-β-cyclodextrin on the physical stability of an otherwise identical co-formulation of caglitide and semaglutide. Composition

The compositions of co-formulations 39 and 40 are shown in Table 36. Table 36 Compositions containing HP-B-CD or SBE-B-CD co-formulations Element Co-formulations 39 40 Caglitide drug substance (mg/ml) 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 - SBE-B-CD (CycloLab) (%w/v) (Average MS: 0.87, MS range: 0.84-0.99) ¹ - 15 L-Histidine (mM) 6 6 Sorbitol(mg/ml) 20 20 Polysorbate 80 (mg/ml) 0.05 0.05 HCl qs qs NaOH qs qs WFI to 1 ml to 1 ml pH 5.8 5.8 ¹ MS: Molar substitution, corresponding to sulfobutylether/hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

Samples used to determine the number of particles visible only under the microscope are stored under pressure conditions, defined as: - Period: 35 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, the samples are inverted for 360° °To simulate the use of patients outside the cold storage room. Three days a week with 20 spins and two days a week with 40 spins. The number of particles visible under the microscope was quantified as described in Example 4. Table 37 Microscopic particle content of caglitinide and semaglutide co-formulations containing HP-B-CD or SBE-B-CD Co-formulations supersede type Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 35 Particle size＞5μm 39 HP-B-CD 49 122 61 245 324 40 SBE-B-CD 317 1686 6940 20714 280478 Particle size＞10μm 39 HP-B-CD 4 8 5 20 20 40 SBE-B-CD 13 640 3503 10248 138324 Particle size＞25μm 39 HP-B-CD 0 1 1 1 4 40 SBE-B-CD 0 126 738 2008 29834 The results for the number of particles visible under the microscope are the average of 3 replicates and have been rounded to the nearest integer value.

The results in Table 37 show that when SBE-B-CD was used to co-formulate caglitinide and semaglutide, a large increase in microscopically visible particles was observed after 14 days; that is to say, the co-formulation was physically is unstable. When HP-B-CD was used instead, little increase was observed during the 35 days of the study; that is, the co-formulation was physically stable.

Contrary to what we have shown for hydroxypropyl-β-cyclodextrin, these results demonstrate that sulfobutoxide-B-cyclodextrin (SBE-B-CD) is not a suitable candidate for coformulation of caglitinide and sima Lutide cyclodextrin. Example 18 : Effect of pH on the physical and chemical stability of co-formulations

This example shows the effect of pH on the physical and chemical stability of caglitide in an otherwise identical co-formulation of caglitide and semaglutide. Composition

The compositions of co-formulations 41 to 45 are shown in Table 38. Table 38 Composition of co-formulations with different pH Element Co-formulations 41 42 43 44 45 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 15 15 15 15 L-Histidine (mM) 6 6 6 6 6 Sorbitol(mg/ml) 20 20 20 20 20 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 0.05 0.05 HCl qs qs qs qs qs NaOH qs qs qs qs qs WFI (ml) amount to 1 amount to 1 amount to 1 amount to 1 amount to 1 pH 5.5 5.6 5.7 5.8 6.0 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

Formulations were prepared as described in Example 1

Samples used to determine the number of particles visible under a microscope are stored under pressure conditions, defined as: - Period: 28 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, samples are inverted 360° to simulate patient use outside the refrigerator. Three days a week with 20 spins and two days a week with 40 spins. The number of particles visible under the microscope was quantified as described in Example 4.

Samples used to determine caglitinide purity were stored at 37°C for up to 28 days. The purity of caglitinide was determined as described in Example 14. Table 39 Physical stability of caglitinide and semaglutide co-formulations with different pH in the pH range of 5.5 to 6.0 Co-formulations pH Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 Particle size＞5μm 41 5.5 70 6594 - 523259 42 5.6 69 174 730 1355 43 5.7 120 117 232 297 44 5.8 49 122 61 245 45 6.0 33 178 133 60 Particle size＞10μm 41 5.5 6 851 - 385284 42 5.6 6 9 200 476 43 5.7 3 5 8 18 44 5.8 4 8 5 20 45 6.0 4 6 3 9 Particle size＞25μm 41 5.5 1 4 - 190781 42 5.6 0 0 18 64 43 5.7 0 0 0 0 44 5.8 0 1 1 1 45 6.0 1 0 0 1 The results for the number of particles visible under the microscope are the average of 3 replicates and have been rounded to the nearest integer value (-). No sampling was performed. Table 40 Cagelintide at different pH values in the pH range 5.5 to 6.0 Chemical purity (%) of caglitide in co-formulation with semaglutide Co-formulations pH Cagelintide purity (%) versus time (days) at 37°C 0 14 28 41 5.5 97.1% 92.1% 85.9% 42 5.6 97.1% 91.6% 85.0% 43 5.7 97.0% 90.6% 83.2% 44 5.8 96.9% 88.9% 81.9% 45 6.0 96.9% 88.2% 78.3% Summary

The results presented in Tables 39 and 40 show that the physical and chemical stabilities of semaglutide and caglitinide depend on the pH of the formulation: the highest pH resulted in the lowest purity of caglitinide at 37°C after 28 days; the lowest pH This results in an increase in the number of particles that can only be seen under a microscope after 14 days. Based on these physical and chemical stability results, the preferred pH range for this particular caglitide and semaglutide co-formulation is 5.6 to 6.0, with a pH of 5.5 not resulting in an acceptable physically stable co-formulation. Example 19 : Effect of the concentration ratio of caglitinide and semaglutide on the physical stability of co-formulations. This example shows that different concentration ratios of caglitinide and semaglutide have an effect on the physical stability of coformulations that can only be seen under a microscope. Effect of particle content on composition

The compositions of histidine buffer co-formulations 46 to 50 are shown in Table 41, and histidine buffer co-formulations 51 to 61 with slightly modified compositions are shown in Table 42. Table 41 Composition of histidine-buffered co-formulations with different concentration ratios of caglitide and semaglutide Element Co-formulations 46 47 48 49 50 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 4.8 6.4 8.0 9.6 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 15 15 15 15 L-Histidine (mM) 6 6 6 6 6 Sorbitol(mg/ml) 20 20 20 20 20 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 0.05 0.05 HCl qs qs qs qs qs NaOH qs qs qs qs qs WFI to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml pH 5.8 5.8 5.8 5.8 5.8 ¹ MS: Molar substitution, corresponding to hydroxypropyl groups per glucose unit Table 42 Composition of histidine buffer co-formulations with modified compositions containing various caglitinide and semaglutide concentration ratios Element Co-formulations 51 52 53 54 55 56 57 58 59 60 61 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 4.8 6.4 8.0 9.6 9.6 9.6 9.6 10.7 12.8 16 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 20 20 20 20 15 20 20 20 20 20 20 L-Histidine (mM) 6 6 6 6 6 6 6 6 6 6 6 Sorbitol(mg/ml) 35 35 35 35 35 20 35 35 35 35 35 Polysorbate 80 (mg/ml) 1.78 1.78 1.78 1.78 0.05 0.05 0.05 1.78 1.78 1.78 1.78 HCl qs qs qs qs qs qs qs qs qs qs qs NaOH qs qs qs qs qs qs qs qs qs qs qs WFI to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml pH 5.8 5.8 5.8 5.8 5.8 5.8 5.8 5.8 5.8 5.8 5.8 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

Samples used to determine the number of particles visible only under the microscope are stored under pressure conditions, defined as: - Period: 28 days - Temperature: 30°C ± 2°C - Pressure conditions: During storage, the samples are inverted for 360° °To simulate the use of patients outside the cold storage room. Three days a week with 20 spins and two days a week with 40 spins. The number of particles visible under the microscope was quantified as described in Example 4. Table 43 Content of particles visible under a microscope in co-formulations containing caglitide and semaglutide at different concentration ratios Co-formulations Caglitide / semaglutide concentration ( ratio ) Number of particles visible under a microscope as a function of time ( days ) 0 14 twenty one 28 Particle size＞5μm 46 3.2 / 3.2 mg/ml (1:1) 207 214 542 341 47 3.2 / 4.8 mg/ml (1:1.5) 65 133 214 591 48 3.2 / 6.4 mg/ml (1:2) 55 76 198 134 49 3.2 / 8.0 mg/ml (1:2.5) 44 86 262 139 50 3.2 / 9.6 mg/ml (1:3) 56 184 137 540 51 3.2 / 3.2 mg/ml (1:1) 297 467 650 11684 52 3.2 / 4.8 mg/ml (1:1.5) 377 541 331 14453 53 3.2 / 6.4 mg/ml (1:2) 588 501 980 45916 54 3.2 / 8.0 mg/ml (1:2.5) 452 341 712 13108 55 3.2 / 9.6 mg/ml (1:3) 37 226 121 161 56 3.2 / 9.6 mg/ml (1:3) 55 77 290 315 57 3.2 / 9.6 mg/ml (1:3) 41 181 273 368 58 3.2 / 9.6 mg/ml (1:3) 312 457 445 25494 59 3.2 / 10.7 mg/ml (1:3.33) 1240 594 1043 53301 60 3.2 / 12.8 mg/ml (1:4) 1459 549 320 77697 61 3.2 / 16 mg/ml (1:5) 1025 568 877 5202 Particle size＞10μm 46 3.2 / 3.2 mg/ml (1:1) 18 31 25 51 47 3.2 / 4.8 mg/ml (1:1.5) 5 9 twenty two 106 48 3.2 / 6.4 mg/ml (1:2) 3 9 18 twenty four 49 3.2 / 8.0 mg/ml (1:2.5) 10 9 13 13 50 3.2 / 9.6 mg/ml (1:3) 1 9 8 130 51 3.2 / 3.2 mg/ml (1:1) 38 45 79 1146 52 3.2 / 4.8 mg/ml (1:1.5) 69 94 80 2053 53 3.2 / 6.4 mg/ml (1:2) 74 81 171 7608 54 3.2 / 8.0 mg/ml (1:2.5) 78 57 118 1651 55 3.2 / 9.6 mg/ml (1:3) 5 9 5 14 56 3.2 / 9.6 mg/ml (1:3) 4 8 twenty two 59 57 3.2 / 9.6 mg/ml (1:3) 0 15 27 61 58 3.2 / 9.6 mg/ml (1:3) 50 79 74 3233 59 3.2 / 10.7 mg/ml (1:3.33) 147 72 165 6793 60 3.2 / 12.8 mg/ml (1:4) 155 84 51 10270 61 3.2 / 16 mg/ml (1:5) 80 99 189 382 Particle size＞25μm 46 3.2 / 3.2 mg/ml (1:1) 1 0 4 1 47 3.2 / 4.8 mg/ml (1:1.5) 0 1 0 6 48 3.2 / 6.4 mg/ml (1:2) 0 1 3 3 49 3.2 / 8.0 mg/ml (1:2.5) 1 0 3 3 50 3.2 / 9.6 mg/ml (1:3) 0 1 0 9 51 3.2 / 3.2 mg/ml (1:1) 0 0 1 13 52 3.2 / 4.8 mg/ml (1:1.5) 4 0 3 60 53 3.2 / 6.4 mg/ml (1:2) 8 4 6 157 54 3.2 / 8.0 mg/ml (1:2.5) 8 3 3 25 55 3.2 / 9.6 mg/ml (1:3) 0 0 1 0 56 3.2 / 9.6 mg/ml (1:3) 3 0 0 3 57 3.2 / 9.6 mg/ml (1:3) 0 0 4 0 58 3.2 / 9.6 mg/ml (1:3) 1 5 0 42 59 3.2 / 10.7 mg/ml (1:3.33) 1 0 5 97 60 3.2 / 12.8 mg/ml (1:4) 3 3 1 68 61 3.2 / 16 mg/ml (1:5) 8 4 26 8 The results for the number of particles visible under the microscope are the average of 3 replicates and have been rounded to the nearest integer value.

The results presented in Table 43 show that after 21 days, few microscopic signs were seen in Co-formulations 46 to 60 containing 3.2 mg/ml caglitide and up to 12 mg/ml semaglutide. Increase in the number of particles.

After 14 days, an increase in the number of microscopic particles was seen for Co-formulation 61 containing 3.2 mg/ml caglitide and 16 mg/ml semaglutide.

All histamine-buffered co-formulations 46 to 61 including 3.2 mg/ml caglitide and up to 16 mg/ml semaglutide were physically stable. Example 20 : Effect of HP-B-CD concentration on physical stability of co-formulations

This example shows the effect of HP-B-CD concentration on the physical stability of caglitide and semaglutide co-formulations where the co-formulations are exposed to physical stress. Composition

The compositions of co-formulations 62 to 65 containing histidine buffer compositions are shown in Table 44. Table 44 Compositions of co-formulations containing different concentrations of HP-B-CD Element Co-formulations 62 63 64 65 Caglitide drug substance (mg/ml) 3.2 3.2 3.2 3.2 Semaglutide drug substance (mg/ml) 3.2 3.2 3.2 3.2 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 7.5 10 12.5 15 L-Histidine (mM) 6 6 6 6 Sorbitol(mg/ml) 20 20 20 20 Polysorbate 80 (mg/ml) 0.05 0.05 0.05 0.05 HCl qs qs qs qs NaOH qs qs qs qs WFI to 1 ml to 1 ml to 1 ml to 1 ml pH 5.8 5.8 5.8 5.8 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulations were prepared as described in Example 1. method

The tendency of caglitinide and semaglutide to aggregate and form peptide fibrils in co-formulations was measured using the thioflavin T (ThT) fluorescence stress assay as described in Example 2. Table 45 Physical stability of caglitide and semaglutide co-formulations with different HP-B-CD concentrations Co-formulations HP-B-CD (MS: 0.62) ² delay time until fibrosis 62 7.5%w/v 17.66 hours ¹ 63 10%w/v 28.06 hours ¹ 64 12.5% w/v 70.97 hours ¹ 65 15%w/v 119.0 hours ^{1 1} result is an average of 6 replicates ² MS: molar substitutions corresponding to hydroxypropyl summary per glucose unit

The results presented in Table 45 show that the physical stability of caglitide and semaglutide co-formulations is dependent on the concentration of HP-B-CD, with lower concentrations resulting in shorter lag times for the onset of fibrosis. The co-formulation including 7.5% w/v HP-B-CD was the least stable. The co-formulation including 15% w/v HP-B-CD was the most stable. Example 21 : Local subcutaneous tolerability of vehicle formulations varying in HP-B-CD content and molar substitution and overall buffer composition when injected subcutaneously

This experiment examines: (1) The effect of HP-B-CD concentration and average MS (0.62 vs. 0.92) on local tolerance profiles after subcutaneous injection. (2) Effect of formulation vehicle on local tolerance profile after subcutaneous injection in the presence of HP-B-CD. Composition

The composition carriers of the co-formulations tested are shown in Table 46. Table 46 Composition of co-formulated vehicles with different HP-B-CD content and molar substitution as well as overall buffer composition Element Co-formulation carrier 9 10 11 12 13 14 15 16 HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.62, MS Range: 0.58-0.68) ¹ 15 20 twenty two twenty two 25 20 twenty two - HP-B-CD KLEPTOSE® (Roquette) (% w/v) (Average MS: 0.92, MS Range: 0.81-0.99) ¹ - - - - - - - twenty two L-Histidine (mM) 6 6 6 6 6 - - - Citrate, 1 H ₂ O (mM) - - - - - 3 3 3 Sorbitol(mg/ml) 20 20 20 35 20 - - - Polysorbate 80 (mg/ml) 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 HCl qs qs qs qs qs qs qs qs NaOH qs qs qs qs qs qs qs qs WFI to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml to 1 ml pH 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 ¹ MS: molar substitution, corresponding to hydroxypropyl process per glucose unit

The formulation was prepared as described in Example 1, except that no active pharmaceutical ingredient was added. method

After subcutaneous administration of formulations containing HP-B-CD, 4 Landrace 4 (32 G/4 mm) to assess skin lesions following subcutaneous administration of 200 µl for 6 days (necropsy) to study local (subcutaneous) tolerance. Skin samples measuring 2x2 cm were collected at necropsy, fixed in neutral buffered formalin, trimmed with a multi-knife, embedded in paraffin, cut into 4 µm thin layers, mounted on glass slides and subsequently Hematoxylin-eosin (HE) staining was performed. Regarding four samples, the severity of subcutaneous tissue necrosis and inflammatory cell infiltration was assessed by trained toxicological pathologists using light microscopy and scored on a numerical scale, where code 1 reflects "no abnormality detected" and code 5 reflects "Significant seriousness": 1. No exception detected 2. Rare severity 3. Mild severity 4. Moderate severity 5. Obvious seriousness

To assess the extent of subcutaneous tissue necrosis and inflammatory cell infiltration induced by co-formulated vehicle when injected subcutaneously. The results are presented in Table 47. Table 47 Subcutaneous tissue necrosis severity scores and inflammatory cell infiltration 6 days after injection of co-formulated vehicles with different HP-B-CD contents and molar substitutions and buffer compositions Co-formulation carrier Necrosis Inflammatory cell infiltration, granulomatous 9 1,1,1,1 2,2,2,2 10 1,1,2,2 2,2,2,3 11 2,3,4,4 3,3,4,4 12 2,4,4,4 3,3,4,4 13 3,4,4,4 3,3,4,5 14 1,3,3,3 1,3,3,3 15 3,4,5,5 4,4,4,5 16 1,5,4,5 2,4,4,5 Summary

The results in Table 47 show that in vivo local subcutaneous tolerance depends on the concentration of HP-B-CD and the overall buffer composition. Co-formulation vehicles including histidine and sorbitol showed better tolerability than those including citrate.

Co-formulated vehicles including 20% w/v HP-B-CD or less resulted primarily in no or minimal necrosis or inflammatory cell infiltration (score 1 or 2) and a single observation of mild inflammatory cell infiltration (score 1 or 2). 3 points). Co-formulated vehicles including 22% w/v or more HP-B-CD resulted in minimal to moderate necrosis and inflammatory cell infiltration (score up to 4). Based on these results, co-formulations containing less than 22% HP-B-CD appear suitable for subcutaneous use.

Co-formulation vehicles containing 20% w/v and 22% w/v HP-B-CD and citrate (co-formulation vehicles 14 and 15) resulted in significant necrosis and inflammatory cell infiltration (score up to 5) . Unexpectedly, co-formulation vehicles containing 20% w/v and 22% w/v HP-B-CD, histidine and sorbitol (co-formulation vehicles 10 and 11) were better tolerated , resulting in moderate necrosis and inflammatory cell infiltration (score as high as 4).

Although certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes and equivalents will now occur to those skilled in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and alterations as fall within the true spirit of the invention.

without

without

Claims (19)

A liquid pharmaceutical formulation comprising an amyloid receptor agonist, a GLP-1 receptor agonist and a hydroxypropyl-substituted alpha or beta cyclodextrin and having a pH value of 5.5 to 6.5, preferably 5.6 to 6.0. The pharmaceutical formulation of claim 1, wherein the GLP-1 receptor agonist is semaglutide. The pharmaceutical formulation according to any one of the preceding claims, wherein the amyloid receptor agonist is caglitinide. A pharmaceutical formulation according to any one of the preceding claims, having a pH value of about 5.6 to 6.0. The pharmaceutical formulation according to any one of the preceding claims, wherein the cyclodextrin is in the beta form substituted with hydroxypropyl. The pharmaceutical formulation of any one of the preceding claims, wherein the cyclodextrin includes a minimum of about 0.4 hydroxypropyl groups per glucose unit and a maximum of about 1.0 hydroxypropyl groups per glucose unit. A pharmaceutical formulation as claimed in any one of the preceding claims, comprising more than 10% w/v and less than 22% w/v cyclodextrin, such as 10% to 20% w/v, such as about 15% w/v cyclodextrin. The pharmaceutical formulation according to any one of the preceding claims, comprising effective amounts of caglitide and semaglutide. The pharmaceutical formulation according to any one of the preceding claims, further comprising a tonicity agent, provided that the tonicity agent is not sodium chloride. The pharmaceutical formulation according to any one of the preceding claims, wherein the tonicity agent is sorbitol, mannitol or trehalose; preferably sorbitol. The pharmaceutical formulation according to any one of the preceding claims, further comprising a buffer having a pKa of at least one of about 5.0 to 7.0, such as citrate or histidine acid; preferably histidine acid. The pharmaceutical formulation according to any one of the preceding claims, further comprising a surfactant such as polysorbate 20 and/or polysorbate 80. The pharmaceutical formulation according to any one of the preceding claims, comprising: - Effective amounts of caglitide and semaglutide, - Hydroxypropyl beta cyclodextrin including a minimum of about 0.4 hydroxypropyl groups per glucose unit and a maximum of about 1.0 hydroxypropyl groups per glucose unit, such as 0.58 to 1.0 hydroxypropyl groups per glucose unit , - Histamine, - Sorbitol, - Polysorbate 20 and/or 80, - Water for injection, and - Has a pH value of 5.6 to 6.0. A pharmaceutical formulation comprising: - Effective amounts of caglitide and semaglutide, - More than 10% w/v and less than 22% w/v, such as 10% to 20% w/v hydroxypropyl beta-cyclodextrin, including a minimum of about 0.4 and a maximum of about 1.0 hydroxyl groups per glucose unit propyl, such as 0.58 to 1.0 hydroxypropyl per glucose unit, - Approximately 3 to 30 mM histamine, - About 10 to 40 mg/ml sorbitol, - Up to 2.0 mg/ml, such as polysorbate 20 and/or 80 up to 1.5 mg/ml, - About 75% to 90% w/w water and - Has a pH value of 5.6 to 6.0. A pharmaceutical formulation comprising: - Effective amounts of caglitide and semaglutide, - More than 10% w/v and less than 22% w/v, such as 10% to 20% w/v hydroxypropyl beta-cyclodextrin, which contains an average of 0.62 to 0.84 hydroxypropyl groups per glucose unit, - Approximately 3 to 30 mM histamine and/or citrate, - About 10 to 40 mg/ml sorbitol, - Up to 2.0 mg/ml, such as polysorbate 20 and/or 80 up to 1.5 mg/ml, - About 75% to 90% w/w water and - Has a pH value of 5.6 to 6.0. A pharmaceutical formulation according to any one of the preceding claims for use as a medicament. The pharmaceutical formulation according to any one of the preceding claims, for the treatment of overweight or obesity with or without one or more related comorbidities; diabetes with or without one or more related comorbidities; heart disease Vascular disease; nonalcoholic steatotic hepatitis (NASH); and cognitive impairment. The pharmaceutical formulation according to any one of claims 1 to 14, for use in any one of claims 16 to 17, characterized in that the formulation is administered by parenteral injection, preferably For subcutaneous injection. The pharmaceutical formulation according to any one of claims 1 to 14 for use in any one of claims 15 to 17, wherein the dose of caglitide administered and the dose of semaglutide administered The dose ratio ranges from 1:1 to 1:7.