Prodrug and its use

Abstract

The present invention relates to DKP-based prodrugs from which a parent drug such as semaglutide is released upon conversion of the prodrug under in vivo conditions. The present invention also relates to the use of the DKP-based prodrugs.

Description

Technical Field

[0001] The present invention relates to DKP-based prodrugs and their therapeutic use.

[0002] Sequence Listing This application is filed with an electronic sequence listing. The entire content of the sequence listing is incorporated herein by reference.

Background Art

[0003] Prodrug technology can be used to generate compounds having properties suitable for a particular dosing frequency. Diketopiperazine (DKP)-based prodrugs have been previously described (e.g., Non-Patent Document 1). This technology is based on a chemical conversion in which a moiety consisting of two amino acids cyclizes to form a six-membered ring and simultaneously releases an active agent.

[0004] Patent Document 1 discloses, according to the claim, a prodrug formulation of a glucagon superfamily peptide in which the peptide is modified by the linkage of dipeptides via amide bond linkages.

[0005] Patent Document 2 discloses, according to the claim, a non-enzymatic self-cleaving dipeptide element linked to a known agent via an amide bond.

[0006] Patent Document 3 discloses, according to the claim, a dipeptide-based prodrug of an aliphatic amine-containing agent.

[0007] Patent Document 4 discloses, according to the claim, a prodrug formulation of a glucagon superfamily peptide in which the peptide is modified by the linkage of dipeptides via amide bond linkages.

[0008] Patent Document 5 discloses, according to the claim, an ester prodrug of an insulin-secreting peptide.

[0009] According to the application, Patent Document 6 discloses a peptide-based prodrug with a significantly extended half-life.

[0010] According to the application, Patent Document 7 discloses a prodrug formulation of insulin and insulin analogs in which the insulin peptide is modified by amide bond linkage of dipeptide prodrug elements.

Prior Art Documents

Patent Documents

[0011]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Non-Patent Documents

[0012]

Non-Patent Document 1

Summary of the Invention

[0013] GLP-1 receptor agonists are widely used in the treatment of chronic diseases. Currently available oral GLP-1 receptor agonists need to be administered once a day. Treatment regimens with less frequent dosing than once a day may provide improved patient convenience and improved patient compliance, and as a result, the development of oral GLP-1 receptor agonists suitable for dosing less frequently than once a day would represent a significant improvement in available treatment options. Prodrug technology may be used to optimize the properties of a drug in a manner suitable for a particular dosing regimen, such as once-weekly dosing. The present invention relates to prodrugs having desirable properties, such as once-weekly oral dosing.

[0014] In a first aspect, the present invention relates to a prodrug of formula I: X-Y-Z, wherein Z is the parent drug and X-Y is a DKP-forming moiety, which prodrug undergoes chemical conversion under in vivo conditions such that the parent drug is released from the DKP-forming moiety. In a second aspect, the present invention relates to a prodrug for use as a medicament. In one functional aspect, the present invention provides a prodrug having a conversion half-life suitable for once-weekly dosing. Additionally or alternatively, in another functional aspect, the present invention provides a prodrug having an observed terminal-phase half-life suitable for once-weekly dosing. Additionally or alternatively, in another functional aspect, the present invention provides a prodrug having surprisingly high oral bioavailability. The present invention may also solve further problems that will become apparent from the disclosure of the exemplary embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0015]

Figure 1

[0016] In the following, Greek letters may be represented by their symbols or corresponding written names, for example, α = alpha, β = beta, γ = gamma, δ = delta, ε = epsilon, ω = omega, etc. Also, the Greek letter μ may be represented by "u", for example, μl is ul and μM is uM. The symbol * in a chemical formula or chemical diagram indicates the bonding point with an adjacent part. In the following, unless otherwise indicated in this specification, terms presented in the singular form also include plural situations. For example, when referring to a "compound", it should be understood that it encompasses all individual variants included within the broad definition of that compound.

[0017] The present invention relates to prodrugs having desirable properties, such as once-weekly oral dosing. In a first aspect, the present invention relates to a prodrug comprising formula I: X-Y-Z (wherein X is an amino acid, Y is selected from the group consisting of Thz and D-Thz, and Z comprises a GLP-1 polypeptide), or a pharmaceutically acceptable salt, ester, or amide of the prodrug. In a second aspect, the present invention relates to the prodrug of the present invention for use as a medicament.

[0018] General Definitions As used herein, the term "compound" refers to a molecular entity. Thus, a "compound" may have different structural elements other than the minimal elements defined for each compound or group of compounds. The term "compound" is used interchangeably with the term "construct". The term "compound" may be used to describe the prodrugs of the present invention. The compounds of the present invention may be referred to as "compounds", and the term "compound" also means encompassing the pharmaceutically relevant forms of this specification, that is, the present invention relates to the compounds defined herein, or their pharmaceutically acceptable salts, amides, or esters.

[0019] As used herein, the terms "polypeptide" or "polypeptide sequence" refer to a compound containing a series of two or more amino acids interconnected via amide (or peptide) bonds. The term polypeptide is used interchangeably with the terms "peptide" and "protein".

[0020] As used herein, the term "analog" generally refers to a polypeptide having one or more amino acid changes in its sequence as compared to a reference amino acid sequence. Such amino acid changes may include amino acid additions, amino acid deletions, and / or amino acid substitutions. Amino acid substitutions, deletions, and / or additions may sometimes also be referred to as "mutations". In certain embodiments, an analog "comprises" a specific change. In other specific embodiments, an analog "consists of" or "has" a specific change. When the terms "comprises" or "comprising" are used with respect to amino acid changes in an analog, it will be understood, of course, that the analog may have additional amino acid changes as compared to its reference sequence. When the terms "consists of" or "has" are used with respect to amino acid changes in an analog, it will be understood, of course, that the specified amino acid mutation is the only amino acid change in the analog as compared to the reference sequence.

[0021] The term "derivative" generally refers to a chemically modified polypeptide in which one or more substituents are covalently attached to the amino acid sequence of the polypeptide, for example, via a bond to the ε-amino group of Lys. In one embodiment, the compounds of the present invention include derivatives that are modified such that one or more substituents having a property of prolonging duration are covalently attached to the amino acid sequence of the polypeptide.

[0022] As used herein, the term "sequence identity" refers to the degree to which two amino acid sequences (e.g., polypeptides) have the same residues at the same positions in an alignment. This may also simply be referred to as "identity". Sequence identity is conveniently expressed as a percentage, i.e., if 85 out of 100 aligned positions between two sequences are identical amino acids, the degree of identity is 85%. For the purposes of the present invention, the sequence identity between two amino acid sequences is determined by using standard protein or peptide alignment programs such as simple handwritten and visual inspection, and / or "align" based on the Needleman-Wunsch algorithm. This algorithm is described in the alignment programs by Needleman, S.B. and Wunsch, C.D., (1970), Journal of Molecular Biology, 48:443-453, and Myers and W. Miller, "Optimal Alignments in Linear Space" CABIOS (computer applications in the biosciences) (1988) 4:11-17. For the alignment, the default score matrix BLOSUM62 and the default identity matrix may be used, the penalty for the first residue in a gap may be set to -12, or preferably -10, and the penalty for additional residues in a gap may be set to -2, or preferably -0.

[0023] Amino acid As used herein, the term "amino acid" refers to any amino acid, i.e., both proteinogenic amino acids and non-proteinogenic amino acids. As used herein, the term "proteinogenic amino acid" refers to the 20 standard amino acids encoded by the genetic code in humans. As used herein, the term "non-proteinogenic amino acid" refers to any amino acid that is not considered a proteinogenic amino acid. Generally, amino acid residues can be specified by, for example, in the context of a polypeptide sequence, when used herein, their full names, their one-letter codes, and / or their three-letter codes when used herein. These three methods are completely equivalent and are used interchangeably. In the following, each amino acid of the peptides of the present invention for which no optical isomers are described should be understood to mean the L-isomer (unless otherwise specified). Examples of non-proteinogenic amino acids incorporated into the compounds of the present invention are listed in Table 1. When something is said to be attached to the "side-chain amino group" of an amino acid, this means attached to the amino group located on the side chain of the amino acid. For example, when a moiety is attached to the side-chain amino group of Lys or D-Lys, this is attached to the ε-amino group; when the moiety is attached to the side-chain amino group of Orn, this is attached to the δ-amino group; and when the moiety is attached to the side-chain amino group of Dab, this is attached to the γ-amino group. [Table 1]

[0024] GLP-1 polypeptide As used herein, the term "GLP-1 polypeptide" refers to a polypeptide that can bind to and / or activate the GLP-1 receptor. In other words, a GLP-1 polypeptide is a polypeptide that is said to have "GLP-1 activity". A GLP-1 polypeptide may bind to and / or activate other types of receptors, i.e., as long as the polypeptide binds to and / or activates the GLP-1 receptor, it is considered a GLP-1 polypeptide regardless of any other receptor interactions it may be involved in. In addition to the amino acid residues involved in GLP-1 receptor interaction, a GLP-1 polypeptide may contain additional amino acid residues that are not involved in GLP-1 receptor interaction.

[0025] As used herein, the term "GLP-1 receptor agonist" refers to a compound that can bind to and / or activate the GLP-1 receptor. In other words, a GLP-1 receptor agonist is said to have "GLP-1 activity". A GLP-1 receptor agonist may be based on any type of molecular scaffold, e.g., small molecule, polypeptide, and antibody, or any combination thereof. A GLP-1 receptor agonist may contain one or more moieties that can activate the GLP-1 receptor.

[0026] As used herein, the term "GLP-1 analog" refers to an analog (or variant) of human glucagon-like peptide-1 (GLP-1(7-37)). The amino acid sequence of human GLP-1(7-37) is included in the Sequence Listing as SEQ ID NO: 1. The amino acid sequence of a GLP-1 analog has one or more amino acid changes compared to GLP-1(7-37). The amino acid changes may include amino acid addition, amino acid deletion, and / or amino acid substitution. The amino acid sequence of semaglutide is a non-limiting example of a GLP-1 analog.

[0027] As used herein, the term "GLP-1 derivative" refers to a chemically modified GLP-1 polypeptide to which one or more substituents are covalently attached to the GLP-1 polypeptide. For example, a GLP-1 derivative is a GLP-1 analog to which one or more substituents are covalently attached. A non-limiting example of a GLP-1 derivative is semaglutide.

[0028] In one embodiment, the compound of the present invention comprises a GLP-1 polypeptide. In one embodiment, the GLP-1 polypeptide is the amino acid sequence of semaglutide. In one embodiment, the compound of the present invention comprises a GLP-1 polypeptide, the GLP-1 polypeptide is a GLP-1 analog, and the GLP-1 analog has a maximum of 3 amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1). In one embodiment, the compound of the present invention comprises a GLP-1 polypeptide, the GLP-1 polypeptide is a GLP-1 analog, and the GLP-1 analog has a maximum of 2 amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1). In one embodiment, the compound of the present invention comprises a GLP-1 derivative, and in a preferred embodiment, the GLP-1 derivative is semaglutide.

[0029] Substituent As used herein, the term "substituent" refers to a moiety that binds to, for example, a GLP-1 polypeptide or a dipeptide extension of a GLP-1 polypeptide, such as the dipeptide extension present in a compound of the present invention, and thus forms part of the DKP-forming moiety, and is covalently bound to a polypeptide. When a substituent binds to a polypeptide or a dipeptide, this polypeptide or dipeptide is said to be "substituted". When a substituent is covalently attached to a polypeptide or an amino acid residue, the polypeptide or amino acid is said to "carry" the substituent. A substituent may comprise a series of individually defined moieties, which may also be referred to as "substituent elements".

[0030] The substituent can form a non-covalent bond with albumin, thereby promoting the circulation of the compound in the bloodstream. Therefore, since the aggregate of the fusion compound and albumin disintegrates only slowly to release the free form of the compound, it may have the effect of prolonging the time the compound exists in the bloodstream. Thus, the substituent may sometimes be referred to as an "albumin-binding moiety" as a whole, and it may be said that the substituent has a "prolonging effect". The substituent may include a portion particularly related to albumin binding and thus prolongation, and this portion may be called a "protractor" or a "prolonging moiety". The substituent may be a lipophilic moiety having a distal carboxylic acid.

[0031] The substituent may include a portion between the prolonging moiety and the point of attachment to the polypeptide, and this portion may be called a "linker". The linker may include several "linker elements". The linker elements can be selected such that they improve the overall properties of the molecule, for example, they improve oral bioavailability, conversion half-life, or the prolonging effect, and thus improve the overall exposure profile upon oral administration of the compound.

[0032] The nomenclature used to describe the prolonging moiety, linker, and other structural elements is as usual in the art. For example, *-CO-* refers to a carbonyl, -CH2- refers to a methylene, -COOH refers to a carboxylic acid, and "-" refers to a covalent bond. Non-limiting examples of substituent elements are listed in Table 2.

Table 2

[0033] As used herein, the term "lipophilic moiety" refers to a moiety containing an aliphatic and / or cyclic hydrocarbon moiety having 6 to 30 carbon atoms, preferably more than 6 and less than 20 carbon atoms. The term "distal carboxylic acid" as used herein in connection with the lipophilic moiety refers to a carboxylic acid that is attached to the most distant (terminal) point of the lipophilic moiety relative to the point of attachment of the lipophilic moiety to an adjacent moiety. For example, in the compounds of the present invention, the lipophilic moiety having a distal carboxylic acid (e.g., Chemical Formulas 1 and 2) is a prolonging moiety, and the carboxylic acid is attached to the most distant (terminal) point of the lipophilic moiety relative to the point of attachment of the lipophilic moiety to an adjacent linker element (e.g., Chemical Formulas 3 and 4). Non-limiting examples of lipophilic moieties having a distal carboxylic acid are Chemical Formulas 1 and 2.

[0034] In one embodiment, the prodrug of the present invention comprises a substituent attached to a dipeptide prodrug moiety. In one embodiment of the present invention, the substituent has a prolonging effect. In one embodiment of the present invention, the substituent comprises a lipophilic moiety having a distal carboxylic acid. In one embodiment of the present invention, the lipophilic moiety having a distal carboxylic acid is selected from the group consisting of Chemical Formulas 1 and 2. In one embodiment, n of Chemical Formula 1 is 12, 14, 16, or 18. In one embodiment, n of Chemical Formula 1 is 14 or 16. In one embodiment of the present invention, the substituent comprises a moiety selected from the group consisting of Chemical Formulas 3 and 4. In one embodiment of the present invention, the substituent is of Formula II: A 5 -A 4 -A 3 -A 2 -A 1 -*(of Formula II). In one embodiment of the present invention, * contributes to the point of attachment to X. In one embodiment of the present invention, A 1 is selected from the group consisting of Chemical Formulas 3, 4, 5, 6, and 7 or is absent. In one embodiment of the present invention, A 2 and A 3 each, individually, is selected from the group consisting of Chemical Formulas 3, 4, and 5 or is absent. In one embodiment of the present invention, A 4is represented by Chemical Formula 3 or Chemical Formula 4. In one embodiment of the present invention, A 5 is selected from the group consisting of Chemical Formula 1 and Chemical Formula 2. In one embodiment of the present invention, the residue A 5 , A 4 , A 3 , A 2 , A 1 are interconnected via an amide bond.

[0035] Prodrug As used herein, the term "prodrug" refers to a compound that undergoes chemical conversion by an enzymatic or non-enzymatic chemical process in vivo to result in the release of the parent drug. As used herein, the term "parent drug" refers to a pharmacologically active compound that is released from the prodrug upon conversion of the prodrug. As used herein, the term "conversion" with respect to a prodrug refers to the process by which the prodrug is converted in an enzymatic or non-enzymatic manner to result in the release of the parent drug. The rate at which conversion occurs can be quantified by the "conversion half-life". The "conversion half-life" is the length of time required to reduce the concentration of the prodrug by half as a result of conversion. The "conversion half-life" may also be referred to as the "prodrug-to-drug conversion half-life" or the "prodrug-to-parent drug conversion half-life".

[0036] An intact prodrug does not exhibit its intended pharmacological activity to a significant extent, for example, not to the extent of being incompatible with a treatment regimen directed at the intended pharmacological activity. When the parent drug is released, the pharmacological activity associated with the intended treatment of the prodrug occurs from the parent drug. When the parent drug is released from the prodrug, it is said to be in its "free form". The prodrug can achieve the desired conversion upon intramolecular cyclization of the terminal dipeptide-based amide extension, at which time the extension is cleaved from the parent drug, resulting in the release of the parent drug in its free form. Such intramolecular cyclization can occur as an enzyme-independent process under physiological conditions, for example, via diketopiperazine (DKP) formation. In a prodrug that is converted via DKP formation, the moiety from which the parent drug is released upon conversion is called the "DKP-forming moiety". The prodrugs of the present invention may have a temporary amide bond between the dipeptide moiety of the DKP-forming moiety and the aliphatic amine group of the parent drug. The conversion half-life can be affected by the structural properties of the DKP-forming moiety. For example, a desired conversion half-life may be obtained by using the dipeptides of the DKP-forming moieties exemplified in the present application. The conversion half-life can be affected by the structural properties of the aliphatic amino acid of the parent drug to which the DKP-forming moiety is linked. For example, a desired conversion half-life may be obtained by using the N-terminal amino acid residue of the parent drug exemplified in the present application. The DKP-forming moiety may be a dipeptide-based extension linked to the parent drug. The DKP-forming moiety may further include structural elements other than the dipeptide, for example, substituents covalently bonded to the dipeptide. The DKP-forming moiety may be inactive or may be associated with pharmacological activity. The conversion of the prodrugs of the present invention occurs mainly in a non-enzymatic manner. In one aspect of the present invention, the prodrugs of the present invention include a DKP-forming moiety.

[0037] Examples of the nomenclature used for the compounds of the present invention that contain a DKP-forming moiety and semaglutide as a parent drug are provided below: N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[10-(3-carboxyphenoxy)decanoylamino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide. In this compound, the DKP-forming moiety contains a Lys residue and a Thz residue. The moiety "[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[10-(3-carboxyphenoxy)decanoylamino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]" is attached to the epsilon amino group of the Lys residue of the DKP-forming moiety. The parent drug is a GLP-1-(7-37) analog in which position 8 is substituted with Aib, position 34 is substituted with Arg, and the moiety "[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]" is attached to the epsilon amino group of the Lys residue at position 26. The complete structure of the compound is shown below.

Chemical formula

[0038] In one embodiment, the compound of the present invention is a prodrug, or a pharmaceutically acceptable salt, ester, or amide thereof. In one embodiment, the compound of the present invention comprises Formula I: X-Y-Z (Formula I). In one embodiment of the present invention, Z is the parent drug. In one embodiment of the present invention, X-Y is a DKP-forming moiety. In one embodiment of the present invention, X is an amino acid. In one embodiment of the present invention, X is selected from the group consisting of Ala, Arg, Asn, Asp, His, Leu, Lys, D-Lys, Phe, Ser, Orn, and Dab. In one embodiment of the present invention, X is an amino acid. In one embodiment of the present invention, X is selected from the group consisting of Lys, D-Lys, Orn, and Dab. In one embodiment of the present invention, X is an amino acid. In one embodiment of the present invention, X is selected from the group consisting of Asp, Lys, and D-Lys. In one embodiment of the present invention, Y is selected from the group consisting of Thz and D-Thz. In one embodiment of the present invention, Z comprises a GLP-1 polypeptide. In one embodiment of the present invention, the N-terminal amino group of the GLP-1 polypeptide is linked to Y via an amide bond. In one embodiment of the present invention, the N-terminal residue of the GLP-1 polypeptide is His. In one embodiment of the present invention, the GLP-1 polypeptide is a GLP-1 analog. In one embodiment of the present invention, the GLP-1 analog has a maximum of 3 amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1). In one embodiment of the present invention, the GLP-1 analog has a maximum of 2 amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1). In one embodiment of the present invention, Z is a GLP-1 derivative. In one embodiment of the present invention, Z is semaglutide. In one embodiment, X optionally bears a substituent, provided that when X bears a substituent, X is selected from the group consisting of Lys, D-Lys, Dab, and Orn. In one embodiment, X is selected from the group consisting of Lys, D-Lys, Dab, and Orn, and X bears a substituent.

[0039] In one embodiment, the compound of the present invention is selected from the group consisting of Chemical Formula 8, Chemical Formula 9, Chemical Formula 10, Chemical Formula 11, Chemical Formula 12, Chemical Formula 13, Chemical Formula 14, Chemical Formula 15, Chemical Formula 16, Chemical Formula 17, Chemical Formula 18, Chemical Formula 19, Chemical Formula 20, Chemical Formula 21, Chemical Formula 22, Chemical Formula 23, Chemical Formula 24, Chemical Formula 25, Chemical Formula 26, Chemical Formula 27, Chemical Formula 28, Chemical Formula 29, Chemical Formula 30, Chemical Formula 31, Chemical Formula 32, Chemical Formula 33, Chemical Formula 34, and Chemical Formula 35, or a pharmaceutically acceptable salt, ester, or amide thereof. In one embodiment, the compound of the present invention is selected from the group consisting of Chemical Formula 8, Chemical Formula 9, Chemical Formula 10, Chemical Formula 11, Chemical Formula 12, Chemical Formula 13, Chemical Formula 14, Chemical Formula 15, Chemical Formula 16, Chemical Formula 17, Chemical Formula 18, Chemical Formula 19, Chemical Formula 20, Chemical Formula 21, Chemical Formula 22, and Chemical Formula 23, or a pharmaceutically acceptable salt, ester, or amide thereof. In one embodiment, the compound of the present invention is Chemical Formula 8. In one embodiment, the compound of the present invention has Chemical Formula 9. In one embodiment, the compound of the present invention has Chemical Formula 10. In one embodiment, the compound of the present invention has Chemical Formula 11. In one embodiment, the compound of the present invention has Chemical Formula 12. In one embodiment, the compound of the present invention has Chemical Formula 13. In one embodiment, the compound of the present invention has Chemical Formula 14. In one embodiment, the compound of the present invention has Chemical Formula 15. In one embodiment, the compound of the present invention has Chemical Formula 16. In one embodiment, the compound of the present invention has Chemical Formula 17. In one embodiment, the compound of the present invention has Chemical Formula 18. In one embodiment, the compound of the present invention has Chemical Formula 19. In one embodiment, the compound of the present invention has Chemical Formula 20. In one embodiment, the compound of the present invention has Chemical Formula 21. In one embodiment, the compound of the present invention has Chemical Formula 22. In one embodiment, the compound of the present invention has Chemical Formula 23. In one embodiment, the compound of the present invention has Chemical Formula 24. In one embodiment, the compound of the present invention has Chemical Formula 25. In one embodiment, the compound of the present invention has Chemical Formula 26. In one embodiment, the compound of the present invention has Chemical Formula 27. In one embodiment, the compound of the present invention has Chemical Formula 28. In one embodiment, the compound of the present invention has Chemical Formula 29. In one embodiment, the compound of the present invention has Chemical Formula 30. In one embodiment, the compound of the present invention has Chemical Formula 31. In one embodiment, the compound of the present invention has Chemical Formula 32. In one embodiment, the compound of the present invention has Chemical Formula 33. In one embodiment, the compound of the present invention has Chemical Formula 34. In one embodiment, the compound of the present invention has Chemical Formula 35.

[0040] Semaglutide Semaglutide is a GLP-1 derivative. Compared with human GLP-1(7-37), semaglutide has substituents covalently linked to the side chains of Aib at position 8, Arg at position 34, and Lys at position 26. The amino acid sequence of semaglutide is included in the Sequence Listing as "[Aib8,Arg34]-GLP-1-(7-37)-peptide" and may be described as such herein. The amino acid sequence of semaglutide is a GLP-1 polypeptide. The amino acid sequence of semaglutide is a GLP-1 receptor agonist. The amino acid sequence of semaglutide is a GLP-1 analog having two amino acid changes compared with human GLP-1(7-37). The amino acid sequence of semaglutide is included in the Sequence Listing as SEQ ID NO: 2.

[0041] The chemical name of semaglutide is N-ε 26 -[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8,Arg34]GLP-1-(7-37).

[0042] Semaglutide has the following structure.

Chemical Structure

[0043] Semaglutide has an elimination half-life of approximately one week in humans. Semaglutide is the active pharmaceutical ingredient of Ozempic®, an injectable prescription drug for adults with type 2 diabetes, and can improve blood glucose levels together with diet and exercise. The dosing frequency of Ozempic® is once a week. Semaglutide is also the active pharmaceutical ingredient of Rybelsus®, an oral prescription drug for adults with type 2 diabetes, and can improve blood glucose levels together with diet and exercise. Rybelsus® is dosed orally once daily as a tablet. An improved patient convenience and patient compliance may be obtained by a treatment regimen with once-weekly oral dosing instead of once-daily oral dosing. The properties of semaglutide are not optimal for once-weekly oral dosing. Semaglutide can be made compatible with once-weekly oral dosing when administered as a suitable prodrug that is converted to semaglutide at a suitable rate upon absorption into the body. Designing such a semaglutide prodrug would constitute a significant improvement in available treatment options. In one embodiment, the parent drug of the prodrug of the present invention is semaglutide.

[0044] Functional properties The therapeutic use of a pharmacologically active compound can be hampered by unsuitable pharmacokinetic properties, for example, because the pharmacokinetic properties are not suitable for reaching the desired exposure after administration of the compound. Prodrug technology can be used to improve the pharmacokinetic properties, for example, to make them suitable for once-weekly oral dosing. The exposure level of the parent drug after administration of the prodrug depends on the conversion half-life from the prodrug to the drug, and thus, by obtaining a suitable conversion half-life, the compound can be made suitable for a particular dosing regimen (e.g., once-weekly dosing). The exposure level of the parent drug after administration of the prodrug depends on the observed terminal-phase half-life of the parent drug, and thus, by obtaining a suitable terminal-phase half-life, the compound can be made suitable for a particular dosing regimen (e.g., once-weekly dosing). The suitability of an orally administered prodrug depends on its ability to reach the systemic circulation after absorption in the gastrointestinal tract, and thus, by obtaining a suitable oral bioavailability, the compound can be made suitable for oral administration (e.g., once-weekly oral administration).

[0045] According to a first functional aspect, the compounds of the invention have a desirable conversion half-life, for example, suitable for once-weekly administration in humans. According to a second functional aspect, the compounds of the invention are associated with a desirable observed terminal-phase half-life of the parent drug, for example, suitable for once-weekly administration in humans. According to a third functional aspect, the compounds of the invention have a desirable oral bioavailability, for example, suitable for oral administration in humans.

[0046] Conversion half-life The rate at which conversion from a prodrug to a drug occurs can be quantified by the conversion half-life. As used herein, the term "conversion half-life" refers to the length of time required to reduce the concentration of the prodrug by half as a result of conversion. A suitable conversion half-life for a semaglutide prodrug intended for once-weekly oral dosing in humans is 3.0 to 21 days when measured in vitro at pH 7.4 and 37°C. A preferred conversion half-life for a semaglutide prodrug intended for once-weekly oral dosing in humans is 3.0 to 14 days when measured in vitro at pH 7.4 and 37°C.

[0047] The prodrug can achieve the desired conversion upon intramolecular cyclization of the terminal dipeptide-based amide extension, whereupon the extension is cleaved from the parent drug, resulting in release of the parent drug in its free form. Such intramolecular cyclization can occur as an enzyme-independent process under physiological conditions, for example, via diketopiperazine (DKP) formation. In prodrugs that are converted via DKP formation, the moiety from which the parent drug is released upon conversion is referred to as the DKP-forming moiety. The conversion half-life depends, inter alia, on the nature of the DKP-forming moiety, and thus, for example, the molecular design of the DKP-forming moiety can be used to improve the conversion half-life (e.g., to make it suitable for once-weekly oral administration) and render the properties of the prodrug suitable for a particular dosing regimen (e.g., once-weekly oral administration).

[0048] The conversion half-life can be measured in vitro, for example, at pH 7.4 and 37°C. The conversion half-life from a prodrug to a drug can be measured as described in common methods for measuring the conversion half-life. In one embodiment, the compound of the invention is a prodrug. In one embodiment of the invention, the prodrug has a conversion half-life from the prodrug to the parent drug of at least 3.0 days when measured in vitro at pH 7.4 and 37°C. In one embodiment of the invention, the prodrug has a conversion half-life from the prodrug to the parent drug of 3.0 to 21 days, preferably 3.0 to 14 days, when measured in vitro at pH 7.4 and 37°C.

[0049] Observed terminal phase half-life Many drugs exhibit a biphasic plasma disposition curve that first follows a steep gradient and then a shallow gradient. The phase following the shallow gradient may be referred to as the "terminal phase". As used herein, the term "terminal phase half-life" refers to the time required for the plasma concentration of a compound to decrease by half during the terminal phase. The terminal phase half-life of a drug when administered in free form is different from that of the drug when administered as a prodrug because continuous release of the drug in free form occurs during in vivo conversion of the prodrug. Thus, the prodrug acts as a depot from which the drug is slowly released. When administered as a prodrug, the terminal phase half-life of the parent drug may also be referred to as the "observed terminal phase half-life". It should be understood that when used with respect to a prodrug, the term "observed terminal phase half-life" refers to the observed terminal phase half-life of the parent drug released during prodrug conversion.

[0050] The observed terminal phase half-life suitable for once-weekly oral administration in humans may be greater than 80 hours, or preferably greater than 90 hours, or most preferably greater than 100 hours when determined in minipigs. The observed terminal phase half-life suitable for once-weekly oral administration in humans may be less than 250 hours, or preferably less than 180 hours when determined in minipigs. The observed terminal phase half-life suitable for once-weekly oral administration in humans may be in the range of 90 to 250 hours, or preferably in the range of 100 to 180 hours when determined in minipigs.

[0051] The observed terminal-phase half-life may be determined in minipigs. The observed terminal-phase half-life can be measured as described in common methods for measuring the terminal-phase half-life. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is more than 80 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is more than 90 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is more than 100 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is more than 110 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is more than 120 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is less than 200 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is less than 190 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is less than 180 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is less than 170 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is less than 160 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is 80 to 200 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is 90 to 180 hours. In one embodiment of the present invention, when the observed terminal-phase half-life of the prodrug of the present invention is determined in minipigs, it is 120 to 160 hours.

[0052] Oral bioavailability Oral treatment with pharmacologically active compounds can be hampered due to poor bioavailability. The term "bioavailability" refers to the ability of a compound to reach the systemic circulation after administration and can be quantified as the fractional extent of the compound dose that reaches the systemic circulation upon administration. It is desirable for a drug for oral administration to have high oral absorbability (i.e., high absorbability from the gastrointestinal tract after oral administration), as high absorbability reduces the dose required to reach the intended systemic concentration of the drug and can thus, for example, reduce the tablet size and manufacturing costs.

[0053] As used herein, the term "oral bioavailability" refers to the ability of a compound to reach the systemic circulation after oral administration. Oral bioavailability reflects the extent to which a compound is absorbed from the gastrointestinal tract after oral administration. In other words, high oral bioavailability is associated with high oral absorbability. High oral bioavailability of a drug is associated with high drug exposure after oral administration. High oral bioavailability of a prodrug is associated with high absorption of the prodrug and results in high exposure of the parent drug after in vivo conversion of the prodrug to the parent drug. Oral bioavailability can be measured, for example, in beagle dogs in a formulation with the absorption enhancer sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC) as described in International Publication No. WO 2019 / 149880.

[0054] Oral bioavailability can be measured as described in common methods for measuring oral bioavailability. In one embodiment, the compounds of the present invention have high oral bioavailability. In one embodiment, the compounds of the present invention have oral bioavailability similar to that of semaglutide. In one embodiment, the compounds of the present invention have oral bioavailability not inferior to that of semaglutide. In one embodiment, the compounds of the present invention have oral bioavailability at least as high as that of semaglutide. In one embodiment, the compounds of the present invention have oral bioavailability suitable for once-weekly oral dosing in humans. In one embodiment, the compounds of the present invention have oral bioavailability determined in beagle dogs and measured as Cmax / dose [kg / L]. In one embodiment, the compounds of the present invention have oral bioavailability determined in beagle dogs and measured as Cmax / dose [kg / L], and Cmax / dose [kg / L] is greater than 0.10, preferably greater than 0.15, preferably greater than 0.20, preferably greater than 0.25, and most preferably greater than 0.30. In one embodiment, the compounds of the present invention have oral bioavailability determined in beagle dogs and measured as AUC / dose [kg*hour / L]. In one embodiment, the compounds of the present invention have oral bioavailability determined in beagle dogs and measured as AUC / dose [kg*hour / L], and AUC / dose [kg*hour / L] is greater than 2.0, preferably greater than 5.0, preferably greater than 10.0, preferably greater than 15.0, and most preferably greater than 20.0.

[0055] GLP-1 activity As used herein, the term "GLP-1 activity" refers to the ability of a compound to activate the GLP-1 receptor. Thus, GLP-1 activity may sometimes be referred to as "GLP-1 potency". GLP-1 activity may be measured as in vitro potency, i.e., performance in a functional GLP-1 receptor assay, more particularly, the ability to stimulate cAMP formation in a cell line expressing the cloned human GLP-1 receptor. GLP-1 activity is EC 50It can be represented as a value. The ability of a compound to bind to the GLP-1 receptor can also be used as a measure of GLP-1 activity. In this case, GLP-1 activity may be referred to as "GLP-1 receptor affinity", and the activity may be expressed as an IC 50 value. Methods for investigating GLP-1 activity are known in the art and are described, for example, in International Publication No. WO 2011 / 073328, International Publication No. WO 2011 / 080102, and International Publication No. WO 2012 / 062803.

[0056] Pharmaceutical efficacy / Medical use The present invention also relates to the compounds of the present invention for use as a medicament. As used herein, the term "treatment" refers to the medical treatment of any human subject in need thereof. Treatment can be preventive, prophylactic, palliative, symptomatic, and / or curative. The timing and purpose of such treatment can vary from individual to individual depending on the health status of the subject.

[0057] In one embodiment, the compounds of the present invention may be used for the treatment and / or prevention of (i) diabetes in any form, (ii) obesity, (iii) non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), (iv) cardiovascular disease, (v) neurodegenerative disorders, (vi) chronic kidney disease (CKD), (vii) diabetic kidney disease (DKD), (viii) peripheral arterial disease (PAD), and / or (ix) heart failure (HF).

[0058] In one embodiment, the present invention relates to a method of treating one or more of (i), (ii), (iii), (iv), (v), (vi), (vii), (viii), and (ix), comprising administering to a patient in need thereof an effective amount of a compound of the present invention, optionally in combination with one or more additional therapeutically active compounds.

[0059] In one embodiment, the compounds of the invention are used for the treatment and / or prevention of all forms of diabetes, such as hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin-dependent diabetes, MODY (maturity-onset diabetes of the young), and gestational diabetes, or for diseases where the reduction of HbA1C is the treatment goal. In one embodiment, the compounds are used for the treatment of cardiovascular diseases, such as syndrome X, atherosclerosis, myocardial infarction, coronary artery disease, reperfusion injury, stroke, cerebral ischemia, early heart disease or early cardiovascular disease, left ventricular hypertrophy, coronary artery disease, hypertension, essential hypertension, acute hypertensive emergency, cardiomyopathy, heart failure, exercise intolerance, acute and / or chronic heart failure, arrhythmia, cardiac dysrhythmia, syncopy, angina pectoris, heart bypass and / or stent restenosis, intermittent claudication (obstructive atherosclerotic arteriosclerosis), diastolic dysfunction, and / or systolic dysfunction; and / or for the treatment of blood pressure reduction, such as reduction of systolic blood pressure. In one embodiment, the compounds are used for the treatment of dyslipidemia and / or diseases where one or more of the following clinical outcomes are the treatment goals: reduction of total serum lipids; increase in HDL; reduction of small dense LDL; reduction of VLDL; reduction of triglycerides; reduction of cholesterol; reduction of plasma levels of lipoprotein a (Lp(a)) in humans; inhibition of the production of apolipoprotein A (apo(A)). In one embodiment, the compounds can be used for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). In one embodiment, the compounds of the invention are used for the treatment and / or prevention of all forms of HF, such as heart failure with reduced ejection fraction (HFrEF), heart failure with mid-range ejection fraction (HFmrEF), and / or heart failure with preserved ejection fraction (HFpEF).

[0060] In one embodiment, the compounds of the invention are used for the treatment of obesity and / or eating disorders where one or more of the following clinical outcomes are the treatment goals: reduction of food intake, increase in energy consumption, reduction of body weight, suppression of appetite, induction of satiety. In one embodiment, the compounds are used for the treatment of neurodegenerative disorders.

[0061] Treatment with the compounds of the present invention can also be combined with one or more additional pharmacologically active substances selected, for example, from cardiovascular therapeutics, anti-diabetic agents, and / or anti-obesity agents. Examples of these pharmacologically active substances are inotropic substances, beta-adrenergic receptor blockers, HMG-CoA reductase inhibitors, angiotensin II receptor antagonists, angiotensin-converting enzyme inhibitors, calcium channel blockers, endothelin antagonists, renin inhibitors, diuretics, aldosterone receptor blockers, endothelin receptor blockers, aldosterone synthase inhibitors, CETP inhibitors, relaxin, PCSK9 inhibitors, BNP and NEP inhibitors, GLP-1 analogs, insulin, sulfonylureas, biguanides, meglitinides, glucosidase inhibitors, glucagon antagonists, DPP-IV inhibitors, SGLT2 inhibitors. Treatment with the compounds of the present invention can also be combined with cardiac surgery.

[0062] Pharmaceutical composition The present invention also relates to a pharmaceutical composition (also referred to as a pharmaceutical formulation) comprising a compound of the present invention. In one embodiment, the pharmaceutical composition comprising the compound comprises at least one pharmaceutically acceptable excipient.

[0063] A pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt, amide, or ester thereof and a pharmaceutically acceptable excipient can be prepared as is known in the art.

[0064] The term "excipient" broadly refers to any component other than the active therapeutic ingredient. An excipient may be an inert substance, a non-active substance, and / or a pharmaceutically inactive substance. Excipients serve various purposes, such as carriers, vehicles, diluents, tablet adjuvants, etc., and / or may function to improve the administration and / or absorption of the active substance. Formulations of pharmaceutically active ingredients with various excipients are known in the art; see, e.g., Remington: The Science and Practice of Pharmacy (e.g., 19th edition (1995) and any subsequent editions). Optional additional components of pharmaceutical compositions include, for example, wetting agents, emulsifiers, antioxidants, bulking agents, metal ions, oily excipients, proteins. Non-limiting examples of excipients are solvents, diluents, buffers, preservatives, isotonic agents, chelating agents, surfactants, and stabilizers.

[0065] The pharmaceutical composition containing the compound of the present invention may be in several dosage forms, such as solutions, suspensions, tablets, and capsules. Preferably, the pharmaceutical composition containing the compound of the present invention is suitable for oral administration. For example, in a preferred embodiment, the pharmaceutical preparation containing the compound of the present invention is prepared in the form of a tablet in which the compound is formulated with the absorption enhancer sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC), as described in, for example, International Publication No. WO 2019 / 149880 or International Publication No. WO 2019 / 215063.

[0066] In a preferred embodiment, the pharmaceutical composition containing the compound of the present invention is used for the same pharmaceutical efficacy as that shown for the compound.

[0067] Manufacturing Process The compounds (or fragments thereof) of the present invention can be prepared by classical peptide synthesis, e.g., solid-phase peptide synthesis using t-Boc or Fmoc chemistry or other well-established techniques, see, e.g., Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999, Florencio Zaragoza Dorwald, "Organic Synthesis on solid Phase", Wiley-VCH Verlag GmbH, 2000, and "Fmoc Solid Phase Peptide Synthesis", Edited by W.C.Chan and P.D.White, Oxford University Press, 2000. Additionally, or alternatively, the compounds (or fragments thereof) can be produced by culturing a host cell that contains a DNA sequence encoding the analog in whole or in part and that can express the peptide in a suitable nutrient medium under conditions that allow for the expression of the peptide. Non-limiting examples of host cells suitable for the expression of these peptides are Escherichia coli, Saccharomyces cerevisiae, and mammalian BHK or CHO cell lines. Those derivatives of the present invention that contain non-coded amino acids may be generated, for example, as described in the experimental part. Or, see, e.g., Hodgson et al: "The synthesis of peptides and proteins containing non-natural amino acids", Chemical Society Reviews, vol.33, no.7 (2004), p. 422-430.

[0068] Specific examples of methods for preparing the derivatives of the present invention are included in the experimental part.

[0069] List of embodiments 1. Formula I: X-Y-Z (Formula I) (wherein X is an amino acid, Y is selected from the group consisting of Thz and D-Thz, Z is a compound comprising a GLP-1 polypeptide, or a pharmaceutically acceptable salt, ester, or amide thereof. 2. The compound according to any of the preceding embodiments, wherein X is selected from the group consisting of Ala, Arg, Asn, Asp, His, Leu, Lys, D-Lys, Phe, Ser, Orn, and Dab. 3. The compound according to any of the preceding embodiments, wherein X is selected from the group consisting of Lys, D-Lys, Orn, and Dab. 4. The compound according to any of the preceding embodiments, wherein X is selected from the group consisting of Lys, Orn, and Dab. 5. The compound according to any of the preceding embodiments, wherein the N-terminal amino group of the GLP-1 polypeptide is linked to Y via an amide bond. 6. The compound according to any of the preceding embodiments, wherein the N-terminal residue of the GLP-1 polypeptide is His. 7. The compound according to any of the preceding embodiments, wherein the GLP-1 polypeptide is a GLP-1 analog. 8. The compound according to any of the preceding embodiments, wherein the GLP-1 polypeptide is a GLP-1 analog, and the GLP-1 analog has a maximum of 3 amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1). 9. The compound according to any of the preceding embodiments, wherein the GLP-1 polypeptide is a GLP-1 analog, and the GLP-1 analog has a maximum of 2 amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1). 10. The compound according to any of the preceding embodiments, wherein Z is a GLP-1 derivative. 11. The compound according to any of the preceding embodiments, wherein Z is semaglutide. 12. The compound according to any of the preceding embodiments, wherein X is linked to Y via an amide bond. 13. A compound according to any of the preceding embodiments, wherein X optionally bears a substituent, provided that when X bears a substituent, X is selected from the group consisting of Lys, D-Lys, Dab, and Orn. 14. A compound according to any of the preceding embodiments, wherein X bears a substituent, provided that when X bears a substituent, X is selected from the group consisting of Lys, D-Lys, Dab, and Orn. 15. A compound according to any of the preceding embodiments, wherein X is selected from the group consisting of Lys, D-Lys, Dab, and Orn and X bears a substituent. 16. A compound according to any of the preceding embodiments, wherein the substituent is attached to Lys via an epsilon-amino group, to D-Lys via an epsilon-amino group, to Dab via a gamma-amino group, or to Orn via a delta-amino group. 17. A compound according to any of the preceding embodiments, wherein the substituent has a long-acting effect. 18. A compound according to any of the preceding embodiments, wherein the substituent contains a long-acting moiety. 19. A compound according to any of the preceding embodiments, wherein the substituent is attached via an amide bond to the side-chain amino group of X. 20. A compound according to any of the preceding embodiments, wherein the substituent contains a lipophilic moiety having a distal carboxylic acid. 21. A compound according to any of the preceding embodiments, wherein the substituent contains a moiety selected from the group consisting of Chemical Formula 1 and Chemical Formula 2. 22. A compound according to any of the preceding embodiments, wherein n in Chemical Formula 1 is 12, 14, 16, or 18. 23. A compound according to any of the preceding embodiments, wherein n in Chemical Formula 1 is 14 or 16. 24. A compound according to any of the preceding embodiments, wherein the substituent contains a moiety selected from the group consisting of Chemical Formula 3 and Chemical Formula 4. 25. The substituent is of formula II: A 5 -A 4 -A 3 -A2 -A 1 -*(Formula II) (wherein * contributes to the bonding point with X, A 1 is selected from the group consisting of Chemical Formula 3, Chemical Formula 4, Chemical Formula 5, Chemical Formula 6, and Chemical Formula 7, or does not exist, A 2 and A 3 each of which is individually selected from the group consisting of Chemical Formula 3, Chemical Formula 4, and Chemical Formula 5, or does not exist, A 4 is Chemical Formula 3 or Chemical Formula 4, A 5 is selected from the group consisting of Chemical Formula 1 and Chemical Formula 2) of the compound according to any one of the preceding embodiments. 26. Residue A 5 , A 4 , A 3 , A 2 , A 1 are interconnected via an amide bond of the compound according to any one of the preceding embodiments. 27. The compound is selected from the group consisting of Chemical Formula 8, Chemical Formula 9, Chemical Formula 10, Chemical Formula 11, Chemical Formula 12, Chemical Formula 13, Chemical Formula 14, Chemical Formula 15, Chemical Formula 16, Chemical Formula 17, Chemical Formula 18, Chemical Formula 19, Chemical Formula 20, Chemical Formula 21, Chemical Formula 22, Chemical Formula 23, Chemical Formula 24, Chemical Formula 25, Chemical Formula 26, Chemical Formula 27, Chemical Formula 28, Chemical Formula 29, Chemical Formula 30, Chemical Formula 31, Chemical Formula 32, Chemical Formula 33, Chemical Formula 34, and Chemical Formula 35 of the compound according to any one of the preceding embodiments, or a pharmaceutically acceptable salt, ester, or amide thereof. 28. The compound is selected from the group consisting of Chemical Formula 8, Chemical Formula 9, Chemical Formula 10, Chemical Formula 11, Chemical Formula 12, Chemical Formula 13, Chemical Formula 14, Chemical Formula 15, Chemical Formula 16, Chemical Formula 17, Chemical Formula 18, Chemical Formula 19, Chemical Formula 20, Chemical Formula 21, Chemical Formula 22, Chemical Formula 23 of the compound according to any one of the preceding embodiments, or a pharmaceutically acceptable salt, ester, or amide thereof. 29. The compound is a prodrug and Z is the parent drug of the compound according to any one of the preceding embodiments. 30. The compound according to any of the preceding embodiments, wherein the compound is a prodrug and X-Y is a DKP-forming moiety. 31. The compound according to any of the preceding embodiments, wherein the compound is a prodrug, Z is the parent drug, and X-Y is a DKP-forming moiety. 32. The compound according to any of the preceding embodiments, wherein the compound has a conversion half-life. 33. The compound according to any of the preceding embodiments, wherein the compound has a conversion half-life suitable for once-weekly dosing. 34. The compound according to any of the preceding embodiments, wherein the compound has a long conversion half-life. 35. The compound according to any of the preceding embodiments, wherein the conversion half-life is measured in vitro at 37 °C and pH 7.4. 36. The compound according to any of the preceding embodiments, wherein the conversion half-life is measured as described in the general method for measuring the conversion half-life. 37. The compound according to any of the preceding embodiments, wherein the conversion half-life is at least 3.0 days. 38. The compound according to any of the preceding embodiments, wherein the conversion half-life is at least 4.0 days. 39. The compound according to any of the preceding embodiments, wherein the conversion half-life is at least 5.0 days. 40. The compound according to any of the preceding embodiments, wherein the conversion half-life is at least 6.0 days. 41. The compound according to any of the preceding embodiments, wherein the conversion half-life is 3.0 to 21 days. 42. The compound according to any of the preceding embodiments, wherein the conversion half-life is 3.0 to 14 days. 43. The compound according to any of the preceding embodiments, wherein the conversion half-life is measured at 37 °C and pH 7.4 and the conversion half-life is 3.0 to 21 days. 44. The compound according to any of the preceding embodiments, wherein the conversion half-life is measured at 37 °C and pH 7.4 and the conversion half-life is 3.0 to 14 days. 45. A compound according to any of the preceding embodiments, wherein the conversion half-life is the conversion half-life from the prodrug to the drug. 46. A compound according to any of the preceding embodiments, wherein the parent drug has a terminal-phase half-life observed upon administration of the prodrug. 47. A compound according to any of the preceding embodiments, wherein the parent drug has a terminal-phase half-life observed upon administration of the prodrug that is suitable for once-weekly dosing. 48. A compound according to any of the preceding embodiments, wherein the parent drug has a long terminal-phase half-life observed upon administration of the prodrug. 49. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life is suitable for once-weekly oral dosing in humans upon administration of the prodrug. 50. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 80 hours. 51. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 90 hours. 52. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 100 hours. 53. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 110 hours. 54. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 120 hours. 55. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 200 hours. 56. A compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined upon administration of the prodrug in minipigs is greater than 190 hours. 57. The compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined at the time of administration of the prodrug in pigs is more than 180 hours. 58. The compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined at the time of administration of the prodrug in pigs is more than 170 hours. 59. The compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined at the time of administration of the prodrug in pigs is more than 160 hours. 60. The compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined at the time of administration of the prodrug in pigs is from 80 to 200 hours. 61. The compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined at the time of administration of the prodrug in pigs is from 90 to 180 hours. 62. The compound according to any of the preceding embodiments, wherein the observed terminal-phase half-life of the parent drug determined at the time of administration of the prodrug in pigs is from 120 to 160 hours. 63. The compound according to any of the preceding embodiments, wherein the compound has oral bioavailability. 64. The compound according to any of the preceding embodiments, wherein the compound has high oral bioavailability. 65. The compound according to any of the preceding embodiments, wherein the compound has oral bioavailability similar to that of semaglutide. 66. The compound according to any of the preceding embodiments, wherein the compound has oral bioavailability not inferior to that of semaglutide. 67. The compound according to any of the preceding embodiments, wherein the compound has oral bioavailability at least as high as that of semaglutide. 68. The compound according to any of the preceding embodiments, wherein the oral bioavailability is suitable for once-weekly oral dosing in humans. 69. A compound according to any of the preceding embodiments, wherein the oral bioavailability is determined in beagle dogs. 70. A compound according to any of the preceding embodiments, wherein the oral bioavailability is determined in beagle dogs upon administration of a tablet comprising 3 mg of the compound, 300 mg of sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (SNAC), and 7.7 mg of magnesium stearate. 71. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L]. 72. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as AUC / dose [kg*hour / L]. 73. A compound according to any of the preceding embodiments, wherein the oral bioavailability is determined as described in a general method for measuring oral bioavailability. 74. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in beagle dogs and Cmax / dose [kg / L] is greater than 0.10. 75. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in beagle dogs and Cmax / dose [kg / L] is greater than 0.15. 76. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in beagle dogs and Cmax / dose [kg / L] is greater than 0.20. 77. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in beagle dogs and Cmax / dose [kg / L] is greater than 0.25. 78. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in beagle dogs and Cmax / dose [kg / L] is greater than 0.30. 79. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as AUC / dose [kg·hour / L] in beagle dogs and the AUC / dose [kg·hour / L] is greater than 2.0. 80. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as AUC / dose [kg·hour / L] in beagle dogs and the AUC / dose [kg·hour / L] is greater than 5.0. 81. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as AUC / dose [kg·hour / L] in beagle dogs and the AUC / dose [kg·hour / L] is greater than 10.0. 82. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as AUC / dose [kg·hour / L] in beagle dogs and the AUC / dose [kg·hour / L] is greater than 15.0. 83. A compound according to any of the preceding embodiments, wherein the oral bioavailability is measured as AUC / dose [kg·hour / L] in beagle dogs and the AUC / dose [kg·hour / L] is greater than 20.0. 84. Semaglutide is N-ε 26 -[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8,Arg34]GLP-1-(7-37), a compound according to any of the preceding embodiments.

Chemical formula

Examples

[0070] This experimental part begins with a list of abbreviations, followed by a section on the general method for preparing the compounds and a section on the method for measuring the properties related to the exposure profile. To illustrate the present invention, several specific examples are incorporated into each of the sections. All the example compounds were prepared according to the general method described herein.

[0071] Abbreviations Aib: α-Aminoisobutyric acid Boc: tert-Butyloxycarbonyl CAD: Charged aerosol detector Dab: 2,4-Diaminobutyric acid DIPEA Diisopropylethylamine DCM: Dichloromethane DIC: Diisopropylcarbodiimide DKP: Diketopiperazine DMF: Dimethylformamide D-PBS Dulbecco's phosphate buffered saline DTT Dithiothreitol EDTA: Ethylenediaminetetraacetic acid Fmoc: 9-Fluorenylmethyloxycarbonyl Ado 8-Amino-3,6-dioxaoctanoic acid HFIP: 1,1,1,3,3,3-Hexafluoro-2-propanol or hexafluoroisopropanol HPLC: High performance liquid chromatography LC: Liquid chromatography LCMS: Liquid chromatography mass spectrometry MeCN Acetonitrile MQ: Milli-Q MS: Mass spectrometry Mtt: 4-Methyltrityl Orn: Ornithine OtBu: tert-Butoxy Oxyma Pure®: Cyano-hydroxyimino-acetic acid ethyl ester Pbf: 2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl PBS Phosphate Buffered Saline RP: Reverse Phase RP-HPLC: Reverse Phase High Performance Liquid Chromatography RT: Room Temperature Sar: Sarcosine SEC: Size Exclusion Chromatography SNAC: Sodium N-(8-(2-hydroxybenzoyl)amino)caprylate SPPS: Solid Phase Peptide Synthesis tBu: tert-Butyl TCEP: Tris(2-carboxyethyl)phosphine TFA: Trifluoroacetic Acid Thz: Thiazolidine-4-carboxylic acid TIPS: Triisopropylsilane Trt: Triphenylmethyl (trityl) UPLC: Ultra Performance Liquid Chromatography UV: Ultraviolet

[0072] General method for preparing the compounds of the present invention In one aspect, the derivatives of the present invention can be prepared as described in the examples herein. In one aspect, the derivatives of the present invention can be prepared as known in the art, i.e., the preparation of peptides can be generated by solid-phase peptide synthesis using classical peptide synthesis, e.g., Boc or Fmoc chemistry or other well-established techniques, see, e.g., Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999, Florencio Zaragoza Dorwald, "Organic Synthesis on solid Phase", Wiley-VCH Verlag GmbH, 2000, and "Fmoc Solid Phase Peptide Synthesis", edited by W.C. Chan and P.D. White, Oxford University Press, 2000.

[0073] Constituent blocks of fatty acids and special amino acids The synthesis of octadecanedioic acid mono-tert-butyl ester was carried out as described in International Publication No. WO 2010 / 102886 (pages 27-28). The corresponding mono-tert-butyl esters of C14, C16, and C20 diacids were prepared as appropriate. The synthesis of 10-(3-tert-butoxycarbonylphenoxy)decanoic acid was carried out as described for 9-(4-tert-butoxycarbonylphenoxy)undecanoic acid in International Publication No. WO 2011 / 080103 (page 131).

[0074] Fmoc-Aib-OH, Boc-Dab(Fmoc)-OH, Fmoc-Glu(OH)-OtBu, Boc-Lys(Fmoc)-OH, Boc-Orn(Fmoc)-OH, Fmoc-Thz-OH, Fmoc-D-Thz-OH were available from Iris Biotech or Sigma-Aldrich.

[0075] Peptide synthesis The preparation of the peptide was carried out using solid-phase peptide synthesis (SPPS) with Fmoc chemistry on a Prelude or Symphony X Solid Phase Peptide Synthesizer from Protein Technologies. The Fmoc-protected amino acids used in this method are the following standard recommended ones: for example, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, and Fmoc-Lys(Mtt)-OH, supplied by, for example, Anaspec, Bachem, Iris Biotech or Novabiochem.

[0076] For the final backbone amino acids, alpha-Boc protected amino acids were used: for example, Boc-Arg(Pbf)-OH, Boc-Asn(Trt)-OH, Boc-Asp(OtBu)-OH, Boc-His(Trt)-OH, Boc-Leu-OH, Boc-Lys(Ac)-OH, Boc-Lys(Boc)-OH, Boc-D-Lys(Boc)-OH, Boc-Phe-OH, Boc-Ser(tBu)-OH, supplied by Bachem, Novabiochem, Iris Biotech or Sigma-Aldrich.

[0077] Wang resin pre-loaded with Fmoc-protected glycine (Fmoc-Gly-Wang) was used. Subsequent amino acids were introduced in a stepwise procedure by a Prelude or Symphony X peptide synthesizer according to the SPPS principle.

[0078] Fmoc deprotection was carried out with 20% piperidine in DMF for 2 × 10 minutes. Introduction of substituents at the alpha position of the N-terminal amino acid was achieved using standard Fmoc-protected amino acids. Peptide coupling was carried out using DIC / Oxyma Pure. The amino acid / Oxyma Pure solution (0.3M / 0.3M in DMF at 3 - 4-fold molar excess) was first added to the resin. Then, the same molar equivalent of DIC was added (0.6M in DMF). The coupling time was 1.5 hours. In some cases, the coupling time was increased or the coupling step was repeated to achieve a sufficient level of coupling. The subsequent capping step was carried out using 1M acetic anhydride in DMF and DIPEA.

[0079] Introduction of substituents at the epsilon nitrogen of lysine at the N-terminus of the sequence was achieved using Boc-Lys(Fmoc)-OH. Introduction of substituents at the delta nitrogen of ornithine at the N-terminus of the sequence was achieved using Boc-Orn(Fmoc)-OH. Introduction of substituents at the gamma nitrogen of 2,4-diaminobutyric acid at the N-terminus of the sequence was achieved using Boc-Dab(Fmoc)-OH.

[0080] For introduction of substituents at the epsilon nitrogen of lysine at position 26, Fmoc-Lys(Mtt)-OH was used. The Mtt group was removed by treatment with HFIP / DCM / TIPS (75:22.5:2.5) (2 × 20 minutes), followed by washing with DCM and DMF, and then the substituent was introduced at the epsilon nitrogen of Lys.

[0081] General cleavage method The peptide was cleaved with TFA / TIPS / H2O / DTT (95:2:2:1) for 2 hours, then the solution was poured into cold diethyl ether and centrifuged. The ether was decanted and the peptide was washed twice with diethyl ether.

[0082] General methods for purification and quantification of derivatives The crude peptide was dissolved in 50% acetic acid in MQ water and purified by reverse-phase preparative HPLC (Waters Delta Prep 4000) on a column containing C18 silica gel. Elution was carried out by increasing the gradient of MeCN in MQ water containing 0.1% TFA. The relevant fractions were analyzed using UPLC. The fractions containing the pure target peptide were pooled. The resulting solution was analyzed (UPLC, LCMS), and the peptide derivative was quantified using a CAD-specific HPLC detector (Vanquish Thermo-Fischer HPLC-CAD). The product was dispensed into glass vials. The vials were capped with a millipore glass fiber pre-filter. By lyophilization, the trifluoroacetate salt of the derivative was obtained as a white solid.

Table 3

[0083] Example 1 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[10-(3-carboxyphenoxy)decanoyl amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0084] Example 2 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{γ}-[(4S)-4-carboxy-4-(15-carboxypentadecanoyl amino) butanoyl] Dab-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0085] Example 3 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{δ}-[(4S)-4-carboxy-4-(15-carboxypentadecanoyl amino) butanoyl] Orn-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0086] Example 4 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino)butanoyl]Lys-D-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0087] Example 5 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0088] Example 6 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{ε}-[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(15-carboxypentadecanoyl amino) butanoyl] amino] butanoyl] amino] butanoyl] Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0089] Example 7 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{ε}-[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino) butanoyl] Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0090] Example 8 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{ε}-[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(15-carboxypentadecanoyl amino) butanoyl] amino] butanoyl] Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0091] Example 9 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{ε}-[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0092] Example 10 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0093] Example 11 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[(2S)-2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]-3-hydroxypropanoyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chemical formula

[0094] Example 12 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0095] Example 13 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]acetyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0096] Example 14 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[2-[[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]amino]acetyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0097] Example 15 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-[(4S)-4-carboxy-4-(19-carboxynonadecanoyl amino)butanoyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0098] Example 16 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-N{ε}-([(2S)-4-carboxy-2-(13-carboxytridecanoylamino)butanoyl]Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0099] Example 17 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-Asp-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0100] Example 18 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0101] Example 19 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-Lys-D-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0102] Example 20 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-D-Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0103] Example 21 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{ε}-(acetyl)Lys-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0104] Example 22 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-Ala-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0105] Example 23 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-Arg-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0106] Example 24 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-Asn-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0107] Example 25 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-His-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0108] Example 26 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-Leu-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0109] Example 27 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-Carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-Phe-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0110] Example 28 N{ε26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-Ser-Thz-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0111] Example 29 - reference compound N{ε 26}-[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoyl amino) butanoyl] amino] ethoxy] ethoxy] acetyl] amino] ethoxy] ethoxy] acetyl]-N{ε}-(octadecanoyl) D-Lys-Sar-[Aib8,Arg34]-GLP-1-(7-37)-peptide

Chem.

[0112] General method for measuring the conversion half-life An assay was performed to investigate the conversion half-life of the prodrug of the prodrug of the present invention to the drug. The conversion half-life was investigated in vitro at pH 7.4 during incubation at 37°C.

[0113] The peptide stock solution was prepared by dissolving the lyophilized powder in PBS buffer to a target of 200 μM. The PBS buffer was gibco 14190 - 094 adjusted to pH = 7.4, Dulbecco's phosphate buffered saline without CaCl 2 and MgCl 2 The pH of the peptide stock solution was adjusted to 7.4 with 0.02 M HCl or 0.02 M NaOH. The samples were filled into Agilent HPLC vials equipped with fixed inserts. The vials were capped to prevent evaporation. The HPLC vials were incubated at 37°C, and the samples were collected at different time points over 2 weeks, rapidly frozen at -80°C, and stored at -20°C until analysis.

[0114] Sample analysis was performed using UPLC combined with UV detection at 215 nm and MS (UPLC - UV - MS). 1 μl of the sample was injected into a Waters Acquity UPLC equipped with a flow - through needle injection system with a particle size of 1.7 μm and maintained at 55°C, and a Waters Acquity CSH C18 column (1 * 150 mm). The flow rate of 100 μl / min was supplied by a binary solvent manager pump containing 0.1% formic acid in water as solvent A and 0.1% formic acid in acetonitrile as solvent B. Gradient elution was performed using 15 - 32% of B from 0 to 4 minutes, followed by 32 - 48% of B from 4 to 54 minutes.

[0115] The identity of the prodrug was confirmed by MS, and the peak purity and area % from the UV signal at 215 nm were plotted as the natural logarithm against time, and the first half-life was calculated using the gradient (k).

Number

[0116] Example 30 The conversion half-life from the prodrug to the drug of the compounds of the present invention was measured as described in the general method for measuring the conversion half-life. The results are presented in Table 3. All compounds of the present invention had a conversion half-life of 3.3 days or more. The compounds of the present invention were associated with a surprisingly high conversion half-life.

Table 4

[0117] General method for measuring the terminal phase half-life The assay was performed to investigate the terminal phase half-life of the drug administered in its free form or the observed terminal phase half-life of the drug administered as a prodrug (i.e., the parent drug). The terminal phase half-life was investigated in minipigs.

[0118] Three Göttingen minipigs (about 25 kg) were equipped with two central catheters. One catheter was used to i.v. dose 10 nmol / kg (0.05 ml / kg) of the test compound formulated in a suitable formulation such as phosphate, propylene glycol, and polysorbate, flushed with physiological saline at pH 7.4. After dosing, blood samples (0.8 ml) were collected via the second catheter at predetermined time points (0 to 3 weeks). The samples were centrifuged and 0.2 ml of plasma was used for bioanalysis.

[0119] Biological analysis was performed as follows: Plasma samples were disrupted by protein precipitation and analyzed by turboflow LCMS. Standard substances were prepared by spiking test compounds into blank plasma from related species, typically in the range of 0.5 - 500 nM. Standard substances, plasma blanks, and study samples were precipitated with 3 volumes of ethanol and then centrifuged at 4°C and 6300 rpm for 30 minutes. In matrices where higher background interference was observed, a second precipitation was performed using acetonitrile in a 1:1 ratio. The supernatant was diluted with water containing 1% formic acid at a ratio of 1:2 (or 1:1). Samples were analyzed by turboflow LCMS using a Cyclone turboflow column (TurboFlow Cyclone 0.5×50 mm, Thermo Fischer Scientific) at room temperature and an Aeris Peptide 3.6μm XB-C18 analytical column (2.1×50 mm, Phenomenex) at 60°C. Gradient elution was used with mobile phase A (consisting of milli-Q water containing 1% formic acid and 5% methanol / acetonitrile (50 / 50)) and mobile phase B (consisting of methanol / acetonitrile (50 / 50) containing 1% formic acid and 5% milli-Q water). A QExactive Plus mass spectrometer was used as the detector in single ion monitoring mode. A linear calibration curve (1 / x2 weighting) was used to calculate the concentration in plasma samples.

[0120] The plasma concentration (vs. time) profile of the test compound was evaluated, and standard pharmacokinetic parameters were estimated by non-compartmental analysis (NCA) using WinNonlin Phoenix 64 (version 8.10, CERTARA). The terminal half-life and / or the observed terminal half-life were estimated using the optimal model that optimizes R 2 The model was constructed to fit the data using the NMLE add-on to Phoenix 64 (version 8.10, CERTARA).

[0121] Example 31 The terminal half-life and / or the observed terminal half-life was measured as described in the general methods for measuring the terminal half-life. The terminal half-life of semaglutide administered in free form was 69 hours in minipigs. The observed terminal half-lives of the two compounds of the present invention and the reference compound are presented in Table 4. The observed (released semaglutide) terminal half-life of the compounds of the present invention was at least 100 hours. The compounds of the present invention are associated with a surprisingly high observed terminal half-life, which constitutes proof of concept of the prodrug technology claimed herein.

Table 5

[0122] General methods for measuring oral bioavailability This assay was performed to measure the oral bioavailability of the compound. The assay determined the exposure of the test compound after oral administration in beagle dogs as described by the relevant pharmacokinetic parameters and plasma concentration curves.

[0123] Preparation of tablets for oral administration: The tablets containing the test compound used in the assay described herein were immediate-release SNAC-based tablets. The test compound was spray-dried as the neutral sodium salt (pH 7-8). Dry granulation was performed by roller compression on a Gerteis MINI-PACTOR. Tablets containing 3 mg of the test compound, 300 mg of sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (SNAC), and 7.7 mg of magnesium stearate were produced on a Kilian Style One using a 7.2×12 mm punch.

[0124] Determination of absorption after oral administration: Eight male beagle dogs aged 1 to 5 years with a body weight of approximately 10 - 12 kg at the start of the study were used. The beagle dogs were housed in groups in enclosures (12 hours light: 12 hours dark) and fed individually and restrictedly once a day with Royal Canin Medium Adult dog (Royal Canin Products, China Branch, or Brogaarden A / S, Denmark). Exercise and social interaction within the group were permitted daily as much as possible. The dogs were used for repeated pharmacokinetic studies with a suitable washout period between consecutive drug administrations. An appropriate acclimation period was given before the start of the first pharmacokinetic study. All handling, dosing, and blood sampling were performed by trained and skilled staff. Before the study, the dogs were fasted overnight and fasted for 0 - 4 hours after dosing. Furthermore, access to water for the dogs was restricted from 1 hour before dosing to 4 hours after dosing, but otherwise, they had free access to water throughout the period.

[0125] Tablets containing the test compound were administered in the following manner: Approximately 3 nmol / kg of SEQ ID NO: 32 was subcutaneously administered to the dogs 10 minutes before tablet administration. The tablet was placed at the back of the dog's mouth to prevent chewing. Then, the mouth was closed, and 10 mL or 50 mL of tap water was given using a syringe to facilitate swallowing of the tablet. Blood was collected at predetermined time points up to 336 hours after dosing to fully cover the plasma concentration - time absorption profile of the prodrug. For each blood sampling time point, approximately 1.2 mL of whole blood was collected into a 1.5 mL EDTA - coated tube, and the tube was gently rotated to mix the sample with EDTA. The blood sample was then maintained on ice until centrifuged (4 minutes, 4°C, 4000 rpm). Plasma was pipetted into microtubes on dry ice and maintained at - 20°C until analysis. Blood samples were collected by syringe from the cephalic vein on the radial side of the forelimb for the first 2 hours, and then from the jugular vein for the remaining time points (the first few drops were discarded from the cephalic vein to prevent heparinized saline from the cephalic vein in the sample).

[0126] Bioanalysis was performed as follows: The plasma concentration of the test compound was assayed by plasma protein precipitation and analyzed by liquid chromatography-mass spectrometry (LC-MS). Standard substances were prepared by spiking blank dog plasma with the analyte to reach a final concentration typically in the range of 2 - 200 nM. Standard substances, plasma blanks, or study samples were prepared for LC-MS by protein precipitation by adding 3 volumes of ethanol followed by centrifugation at 4000 rpm for 1 hour at 4°C. The supernatant was diluted with 2 volumes of Milli-Q water containing 1% formic acid and then injected into the LC-MS system. The system used was a Transcend II Interface Module SRD3200 system from Thermo Scientific (Waltham, MA, USA) coupled with an Orbitrap Exploris 240 mass spectrometer from Thermo Scientific. LC was equipped with a Cyclone column (CH-953288, Thermo Scientific) as a one-dimensional capture column and a Poroshell 120 SB-C18 2.7 μm (2.1×50 mm, from Agilent, Santa Clara, CA, USA) as an analytical column. The composition of the mobile phase for the load pump was as follows: Mobile phase A consisted of 95% milli-Q water, 2.5% acetonitrile, 2.5% methanol, and 0.1% formic acid. Mobile phase B consisted of 47.5% acetonitrile, 47.5% methanol, 5% milli-Q water, and 0.1% formic acid. The analyte of interest was loaded from the Turbo flow column to the two-dimensional analytical column at 30% B. Gradient elution was performed with the elution pump using mobile phase A (95% milli-Q water, 2.5% acetonitrile, 2.5% methanol, and 0.1% formic acid) and mobile phase B (47.5% acetonitrile, 47.5% methanol, 5% milli-Q water, and 0.1% formic acid) with a gradient of 0% mobile phase B to 70% mobile phase B in 0.25 minutes, 70% mobile phase B to 80% mobile phase B in 1.17 minutes, and then 80% mobile phase B to 95% mobile phase B in 1.17 minutes.The Orbitrap Exploris 240 was operated in positive ionization mode with parallel reaction monitoring (PRM) scan mode. A linear calibration curve (1 / x² weighted) was used to calculate the test compound concentration in the plasma sample and to determine the maximum plasma concentration (Cmax). Quality control samples for the analyte were included. The deviation between the nominal and calculated concentrations in the standards and quality control samples was below 15%, and for the LLOQ samples it was below 20%. The plasma concentration (vs. time) profile of the test compound was evaluated, and the standard pharmacokinetic parameters were estimated by non-compartmental analysis (NCA) using WinNonlin Phoenix 64 (version 8.10, CERTARA). The results were reported as dose-corrected plasma concentration (vs. time) profiles, as well as dose-corrected maximum plasma concentration (Cmax / dose) and area under the curve (AUC / dose).

[0127] Example 32 Oral bioavailability was determined as described in the general method for measuring oral bioavailability. The relevant pharmacokinetic parameters of two compounds of the present invention and a reference compound are presented in Table 5. The compounds of the present invention were associated with surprisingly high Cmax / dose. The dose-corrected plasma concentration (vs. time) profiles of two compounds of the present invention and the reference compound are presented in Figure 1. The compounds of the present invention were associated with surprisingly high exposure as determined by the dose-normalized plasma concentration profile. The compounds of the present invention were associated with surprisingly high oral bioavailability. [Table 6]

Claims

1. Formula I: X-Y-Z (Equation I) (In the formula, X is an amino acid, Y is selected from the group consisting of Thz and D-Thz. Z is a compound containing GLP-1 polypeptide. or a pharmaceutically acceptable salt, ester, or amide thereof.

2. The compound according to claim 1, wherein X is selected from the group consisting of Ala, Arg, Asn, Asp, His, Leu, Lys, D-Lys, Phe, Ser, Orn, and Dab.

3. The compound according to claim 1, wherein the N-terminal amino group of the GLP-1 polypeptide is linked to Y via an amide bond.

4. The compound according to claim 1, wherein the N-terminal residue of the GLP-1 polypeptide is His.

5. The compound according to claim 1, wherein the GLP-1 polypeptide is a GLP-1 analog having up to two amino acid changes compared to GLP-1(7-37) (SEQ ID NO: 1).

6. The compound according to claim 1, wherein Z is semaglutide.

7. The compound according to claim 1, wherein X supports a substituent, provided that if X supports a substituent, X is selected from the group consisting of Lys, D-Lys, Dab, and Orn.

8. The compound according to claim 7, wherein the substituent comprises a lipophilic moiety having a distal carboxylic acid.

9. The lipophilic portion having the distal carboxylic acid is defined by chemical formula 1 or chemical formula 2: 【Chemistry 1】 The compound according to claim 8.

10. The substituents are chemical formulas 3 and 4: 【Chemistry 2】 The compound according to claim 7, comprising a portion selected from the group consisting of the following.

11. The substituents are of formula II: A ５ -A ４ -A ３ -A ２ -A １ -*(Formula II) (In the formula, * provides a point of connection with X, A １ However, chemical formulas 3, 4, 5, 6, and 7: 【Transformation 3】 Selected from the group consisting of, or if none exist, A ２ and A ３ Each of these is individually selected from the group consisting of chemical formula 3, chemical formula 4, and chemical formula 5, or none exists. A ４ is represented by Chemical Formula 3 or Chemical Formula 4, A ５ However, chemical formulas 1 and 2: 【Chemistry 4】 The compound according to claim 7, which is selected from the group consisting of the following.

12. The aforementioned fat A ５ A ４ A ３ A ２ A １ The compound according to claim 11, wherein the compounds are interconnected via amide bonds.

13. The compound according to claim 1, wherein the compound is selected from the group consisting of chemical formulas 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, and 35, or is a pharmaceutically acceptable salt, ester, or amide thereof.

14. A pharmaceutical product comprising the compound described in any one of claims 1 to 13.

15. The pharmaceutically acceptable agent according to claim 14 for use in the treatment of (i) diabetes mellitus, (ii) obesity, (iii) non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), (iv) cardiovascular disease, (v) neurodegenerative disorders, (vi) chronic kidney disease (CKD), (vii) diabetic kidney disease (DKD), (viiii) peripheral artery disease (PAD), and / or (ix) heart failure (HF).

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