MASP-INHIBITORY COMPOUNDS AND USES THEREOF

Abstract

The present invention relates to novel mannose-binding lectin (MBL)-associated serine protease (MASP) inhibitory compounds, as well as analogues and derivatives thereof, methods for their preparation, their use alone or in combination for the treatment and / or prevention of diseases, and their use for the manufacture of medicaments for the treatment and / or prevention of diseases, particularly renal and cardiovascular and ischemia-reperfusion injury.

Description

[Technical Field]

[0001] The present invention relates to novel mannose-binding lectin (MBL)-associated serine protease (MASP) inhibitory compounds, as well as analogues and derivatives thereof, processes for their preparation, their use alone or in combination for the treatment and / or prevention of diseases, and their use for the manufacture of medicaments for the treatment and / or prevention of diseases, in particular for the treatment and / or prevention of renal and cardiovascular disorders and ischemia-reperfusion injury.

[0002] The complement system consists of a complex cascade network of proteins, receptors, and enzymes, many of which circulate in the bloodstream. It is a key component of innate immunity and is essential for defense against invading pathogens and clearance of dead and virus-infected cells. It forms a bridge between the innate and adaptive immune responses. Complement activation is also involved in the pathology of sepsis and ischemia-reperfusion injury, such as those following myocardial infarction, ischemic kidney injury, or organ transplantation. Three branches of the complement system have been identified: the lectin pathway, the classical pathway, and the alternative pathway (Dunkelberger and Song, Complement and its role in innate and adaptive immune responses. Cell Res. 2010;20(1):34-50). The lectin pathway is activated by the deposition of lectins circulating in the bloodstream. Under normal conditions, the lectin pathway recognizes and modifies foreign and modified carbohydrate surface patterns, respectively, thereby marking invading pathogens and dead cells. Mannose-binding lectin (MBL), ficolins, and collectins are the major representatives of these lectins produced in the liver, kidney, and other organs (Garred et al., "The Complement-MBL and Subsequent Journey Through the Mannose-Binding Lectin Pathway," Immunol Rev. 2016;274(1):74-97). Following their deposition, the zymogens of two closely related serine proteases, mannose-binding lectin-associated serine proteases 1 and 2 (MASP-1 and MASP-2), are recruited from the bloodstream, forming a complex in which the zymogens are in close proximity to each other. The current concept is that under in vivo conditions, the recruited MASP-1 zymogen autoactivates and then activates the MASP-2 zymogen by cleavage. Activated MASP-1 further cleaves complement factor C2 into C2a and C2b. In addition, activated MASP-2 cleaves C2 and complement factor C4 into C4a and C4b, and together with C2a forms the C4bC2a complex, which acts as a complement factor C3 convertase.The formation of C3 convertase activity and subsequent deposition of C3 on the target cell surface represent a convergence point for all three complement pathways, activating a common downstream cascade that leads to the generation of inflammatory mediators and target cell lysis. In intact human serum, the activity of both MASP enzymes is essential for C3 convertase formation (Heja et al., A revised mechanism of complement lectin pathway activation revealing a role for the serine protease MASP-1 as the exclusive activator of MASP-2, Proc Natl Acad Sci USA. 2012;109(26):10498-503).

[0003] The microvasculature plays a crucial role during inflammatory and ischemic organ injury. Barrier function, leukocyte trafficking, and coagulation control are closely dependent on the integrity of the luminal endothelial cell surface in small blood vessels. The luminal endothelial surface is covered throughout by a dense coat of glycosylated extensions consisting of membrane-integrated glycoproteins, proteoglycans, and glycolipids, known as the glycocalyx. Electron microscopic analysis of specimens from animal studies and human pathology has shown that the endothelial glycocalyx, in particular, is rapidly and drastically degraded during ischemic stress as well as under inflammatory conditions such as sepsis. These changes result in the exposure of carbohydrate residues to the bloodstream that are undetectable under normal conditions (for a review, see Sieve et al., Regulation and Function of the Endothelial Glycocalyx in Vascular Disease, Vascul Pharmacol, 2018;100:26-33). This altered carbohydrate pattern, in addition to other changes in the cell surface, is thought to activate the lectin pathway, which triggers the deposition of lectin-recognizing patterns, subsequent C3 deposition, and the initiation of cell lysis. MBL and C3 deposition has been shown to occur after ischemia and acute kidney injury across species, including humans. Lectin pathway activation was particularly relevant to reperfusion injury, as targeted deletion of MBL and MASP-2 protected mice from ischemia-reperfusion injury in the kidney, heart, and intestine (Moller-Kristensen et al., Mannan-binding lectin recognizes structures on the ischemic-reperfused mouse kidney and is involved in tissue damage, Scand J Immunol 2005;61(5):426-34; Schwaeble et al., Targeting mannan-binding lectin-associated serine protease-2 protects against myocardial and gastrointestinal ischemia / reperfusion injury, Proc Natl Acad Sci U S A. 2011;108(18):7523-8). Furthermore, deletion of collectin-11, another MASP-activating lectin primarily expressed in the kidney, produced mice resistant to ischemic acute kidney injury (Farrar et al., Collectin-11 detects stress-induced L-fucose patterns and induces renal epithelial injury, J Clin Invest, 2016;126(5):1911-1925).Selective peptide inhibitors of MASP-1 and MASP-2 have been identified from phage display libraries using natural trypsin inhibitors from sunflower or grasshopper as a starting point. These peptides have been shown to inhibit lectin pathway-dependent C3 convertase formation in vitro (Kocsis et al., "Selective inhibition of the lectin pathway of complement by phage display displays selected peptides against mannose-binding lectin-associated serine proteases (MASP)-1 and -2: significant contribution of MASP-1 to lectin pathway activation," J Immunol, 2010;185(7):4169-78; Heja et al., "Monospecific inhibitors demonstrate that both mannan-binding lectin-associated serine protease-1 (MASP-1) and -2 are essential for lectin pathway activation and reveal structural plasticity of MASP-2," J Biol Chem, 2012;287(24):20290-300). However, evidence of the pharmaceutical usefulness and in vivo efficacy of these peptide inhibitors is not yet available. Similarly, antibodies against MASP-2 that interfere with MASP zymogen interaction have been identified and are in clinical development for atypical hemolytic uremic syndrome and other inflammatory kidney disorders (ClinicalTrials.gov Identifier: NCT03205995; NCT02682407; NCT03608033). However, clinical evidence of their usefulness in the prevention or treatment of acute organ damage, especially ischemic organ damage, is still lacking.

[0004] WO2004 / 075837 discloses anti-MASP antibodies, functionally equivalent fragments thereof, and MASP-binding peptides for reducing morbidity and mortality caused by tissue damage associated with ischemia-reperfusion injury or TAAA repair through inhibition of the complement system. Small peptides such as sunflower MASP inhibitor-1 (SFMI-1) and sunflower MASP inhibitor-2 (SFMI-2) and their derivatives for the treatment of diseases associated with the complement system, primarily the lectin pathway, were first described in WO2010 / 136831.

[0005] WO2015 / 054298 discloses methods for preserving vision or reducing vision loss in a subject by reducing the activity of MASP-1, MASP-2, or MASP-3, and methods for inhibiting or reducing photoreceptor cell death in a subject. WO2004 / 106384, WO2005 / 123128, WO2007 / 117996, and WO2014 / 144542 disclose anti-MASP-2 antibodies for the treatment of diseases associated with MASP-2-dependent complement activation. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] WO2004 / 075837 [Patent Document 2] WO2010 / 136831 [Patent Document 3] WO2015 / 054298 [Patent Document 4] WO2004 / 106384 [Patent Document 5] WO2005 / 123128 [Patent Document 6] WO2007 / 117996 [Patent Document 7] WO2014 / 144542 [Non-patent literature]

[0007] [Non-Patent Document 1] Garred et al., Complement-MBL and Beyond: A Journey Through the Mannose-Binding Lectin Pathway, Immunol Rev. 2016;274(1):74-97 [Non-patent document 2] Heja et al., A revised mechanism of complement lectin pathway activation revealing a role for the serine protease MASP-1 as the exclusive activator of MASP-2, Proc Natl Acad Sci USA. 2012;109(26):10498-503 [Non-patent document 3] Sieve et al., Regulation and function of the endothelial glycocalyx in vascular disease, Vascul Pharmacol, 2018;100:26-33 [Non-patent document 4] Moller-Kristensen et al., Mannan-binding lectin recognizes structures on the ischemic-reperfused mouse kidney and is involved in tissue injury, Scand J Immunol, 2005;61(5):426-34 [Non-Patent Document 5] Schwaeble et al., Targeting mannan-binding lectin-associated serine protease-2 confers protection from myocardial and gastrointestinal ischemia / reperfusion injury, Proc Natl Acad Sci U S A. 2011;108(18):7523-8 [Non-patent document 6] Farrar et al., Collectin-11 detects stress-induced L-fucose patterns and induces renal epithelial injury, J Clin Invest, 2016;126(5):1911-1925 [Non-Patent Document 7] Kocsis et al., Selective inhibition of the lectin pathway of complement by phage display selected peptides against mannose-binding lectin-associated serine proteases (MASP)-1 and -2: significant contribution of MASP-1 to lectin pathway activation, J Immunol, 2010;185(7):4169-78 [Non-patent document 8] Heja et al., Monospecific inhibitors demonstrate that both mannan-binding lectin-associated serine protease-1 (MASP-1) are essential for lectin pathway activation and reveal structural plasticity of MASP-2, J Biol Chem, 2012;287(24):20290-300 Summary of the Invention [Problem to be solved by the invention]

[0008] The object of the present invention was to provide novel peptides having an inhibitory effect on the MASP-1 and / or MASP-2 enzymes and other beneficial properties, making them suitable as an efficient and safe alternative for the prevention and / or treatment of MASP-1 and / or MASP-2-related disorders as defined below. A further object was to provide novel peptides having an improved inhibitory effect on human MASP-1 and / or MASP-2 enzymes and / or rat MASP-1 and / or MASP-2 enzymes. [Means for solving the problem]

[0009] The present invention relates generally to peptides that act as inhibitors of the MASP-1 and / or MASP-2 enzymes, and methods for making and using them.

[0010] In particular, the present invention provides compounds of the following structural formula (I): [ka]

[0011] [During the ceremony, X 0 is represented by formula (IIa) [ka]

[0012] [where: * indicates a bond to the terminal amino group of the adjacent amino acid; A is a bond or C1-C6-alkylene, in which one CH2 group may be replaced by -O- or -S-, and in which the C1-C6-alkylene is up to three-substituted by identical or different radicals selected from the group consisting of hydroxy, methoxy, ethoxy, carboxy, amino and halogen; B is absent, aryl, heteroaryl, C3-C8-cycloalkyl or C3-C7-heterocycloalkyl, wherein aryl, heteroaryl, C3-C8-cycloalkyl and C3-C7-heterocycloalkyl can be up to three-substituted by identical or different radicals selected from the group C1-C4-alkyl, hydroxyl, methoxy, ethoxy, carbonyl, carboxy, amino and halogen, R 1 is hydrogen, halogen, amino, hydroxyl or C1-C 20 -alkyl, where C1-C 20 - alkyl is up to trisubstituted with identical or different radicals selected from the group consisting of hydroxyl, carboxy, amino and halogen. represents a group of p represents an integer of 0 or 1; X 1 represents any natural or unnatural amino acid, where any natural and / or unnatural amino acid can be in the D- or L-configuration; Also, if p is 0 and q is different from 0, then X 1 The terminal amino group of is unsubstituted, acetylated, or C1-C 20 -alkyl mono- or di-substituted, q represents an integer from 0 to 5, X 2 represents a natural amino acid selected from the list consisting of I, L, M, V and A, or a non-natural amino acid selected from the list consisting of LN-methylisoleucine ((N-Me)I), allo-L-isoleucine (allo-I), L-cyclobutylalanine (Cba), L-norvaline (Nva), L-2-aminobutyric acid (Abu), (2S,3S)-2-[(3R)-3-amino-2-oxopyrrolidin-1-yl]-3-methylpentanoic acid, (2S,3S)-2-[2-oxopiperazin-1-yl]-3-methylpentanoic acid, (2S,3S)-2-[(3S)-2-oxopiperazin-1-yl]-3-methylpentanoic acid, L-methionine-L-sulfoxide, L-methionine-sulfone and L-tert-butylglycine, On the other hand, each of the natural amino acids and / or unnatural amino acids in the L-configuration can be replaced with its stereoisomer in the D-configuration; And if p and q are both 0 and r is 1, then X 2 The terminal amino group of is unsubstituted, acetylated, or C1-C 20 -alkyl mono- or di-substituted, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of L-penicillamine (Pen) and LN-methylcysteine ​​((N-Me)C), And if p, q, and r are all 0, then X 3 The terminal amino group of is unsubstituted, acetylated, or C1-C 20 -alkyl mono- or di-substituted, X 4 represents a natural amino acid selected from the list consisting of S, C, T, R or K, or represents an unnatural amino acid selected from the list consisting of allo-L-threonine (allo-T), L-homoserine (hSer) and L-ornithine (Orn); X 5 represents the natural amino acid R or N(5)-methyl-L-arginine ((Me)R), X 6 represents a natural amino acid selected from the list consisting of S, C, or T, or an unnatural amino acid selected from the list consisting of allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap), X 7represents a natural amino acid selected from the list consisting of L, F or N, or is selected from the group consisting of 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), L-4-bromophenylalanine ((4-bromo)F), 2,5-difluoro-L-phenylalanine ((2,5-difluoro)F), L-tert-butylalanine ((tBu)A), 2-chloro-L-phenylalanine ((2-chloro)F) , L-2-Bromophenylalanine ((2-bromo)F), (S)-2-(amino)-1,6-hexanedioic acid (AAD), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-2-amino-4-cyanobutyric acid (Cnba), 4-fluoro-leucine ((4-fluoro)L), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propanoic acid, L-tert- Butylglycine ((tBu)G), 3-(trimethylsilyl)-L-alanine, 2,5-difluoro-L-phenylalanine, 2-amino-7-(tert-butoxy)-7-oxoheptanoic acid, 5,5,5-trifluoro-L-leucine ((trifluoro)L), 2-methyl-L-phenylalanine ((2-Me)F), L-cyclobutylalanine (Cba), L-cyclopentylalanine (Cpa), L-cyclopropylmethylalanine (CpG), represents an unnatural amino acid selected from the list consisting of phenylalanine, L-trifluoromethylalanine, L-difluoromethylalanine, 2-fluoro-L-phenylalanine ((2-fluoro)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, (2S)-3-(3-cyanophenyl)-2-aminopropanoic acid, and (2S)-3-(indol-4-yl)-2-(amino)propanoic acid; X 8represents the natural amino acid P or an unnatural amino acid selected from the list consisting of (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, L-hydroxyproline (Hyp), (3S)-morpholine-3-carboxylic acid (morpholine-3-carboxylic acid), L-pipecolic acid (Pip), (4aR,6aR,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid, trans-4-fluoroproline ((trans-4-fluoro)P) and (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid; X 9represents the natural amino acid P, or is an amino acid selected from the group consisting of 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), L-hydroxyproline (Hyp), (2S,4S)-4-trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,4S)-4-fluoroproline ((cis-4-fluoro)P), trans-4-fluoroproline ((trans-4-fluoro)P), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-trans-3-hydroxyproline ((3S-OH)P, (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-propanol, 2-( ... (2R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1) and rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2), X 10 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of L-cyclopentylglycine (Cpg), L-cyclohexylglycine (Chg), (S)-2-amino-3-ethyl-pentanoic acid, 3-chlorophenylglycine ((3-chloro-Ph)G), L-tert-butylglycine, allo-L-isoleucine (allo-I), L-cyclobutylglycine, L-norvaline (Nva), and (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid; X 11represents the natural amino acid C or an unnatural amino acid selected from the list consisting of L-N-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of allo-L-isoleucine (allo-I), (S)-2-amino-2-cyclobutylacetic acid (Cbg), (2S,3S)-2-((amino)methyl)-3-methylpentanoic acid, L-phenylglycine (Phg), 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, 2-methyl-D-alloisoleucine, L-norvaline (Nva), L-2-aminobutyric acid (Abu), L-tert-butylglycine, and aminoisobutyric acid (Aib); And when u, t, and s are all 0, X 12 the terminal carboxyl group of which is unsubstituted or amidated, X 13 represents a natural amino acid selected from the list consisting of P, A, S, T, G, D, E, Q or N, or N-methyl-glycine ((N-Me)G), 5-azaspiro[2.4]heptane-6-carboxylic acid, L-2-aminobutyric acid (Abu), 2-aminoisobutyric acid (Aib), 2-methyl-L-proline (2-Me)P, hydroxyproline (Hyp), 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), trans-4-fluoroproline ((trans-4-fluoro)P), (2S,4S)-4-fluoroproline ((cis-4-fluoro)P), L-4,4-difluoroproline ((difluoro)P), L-cyclopentylglycine (Cpg), (S)-2-amino-2-cyclobutylacetic acid (Cbg), and (2S)-pyrrolidin-2-ylacetic acid (beta-homo-P), On the other hand, each of the natural amino acids and / or unnatural amino acids in the L-configuration can be replaced with its stereoisomer in the D-configuration; And when u and t are both 0 and s is different from 0, then X 13the terminal carboxyl group of which is unsubstituted or amidated, s represents an integer from 0 to 3, X 14 represents any natural or unnatural amino acid, while any natural and / or unnatural amino acid can be in the D- or L-configuration; And when u is 0 and t is different from 0, X 14 the terminal carboxyl group of which is unsubstituted or amidated, t represents an integer from 0 to 4, X 15 is represented by formula (IIb) [ka]

[0013] [where: * indicates a bond to the terminal carboxyl group of the adjacent amino acid; A 1 is a bond or C1-C6-alkylene, in which one CH2 group may be replaced by -O- or -S-, and the C1-C6-alkylene is up to three-substituted by identical or different radicals selected from the group consisting of hydroxy, methoxy, ethoxy, carboxy, amino and halogen, B 1 is absent or is aryl, heteroaryl, C3-C8-cycloalkyl or C3-C7-heterocycloalkyl, wherein aryl, heteroaryl, C3-C8-cycloalkyl and C3-C7-heterocycloalkyl can be up to three-substituted by identical or different radicals selected from the group C1-C4-alkyl, hydroxyl, methoxy, ethoxy, carbonyl, carboxy, amino and halogen, and R 2 is hydrogen, halogen, amino, hydroxyl or C1-C 20 -alkyl, where C1-C 20- alkyl is up to trisubstituted with identical or different radicals selected from the group consisting of hydroxyl, carboxy, amino and halogen. represents a group of u represents an integer of 0 or 1; However, X 1 ~X 14 at least one of which is an unnatural amino acid. The present invention provides a compound comprising the peptide or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of the salt, which comprises, consists essentially of, or consists of, and can be isolated and / or purified.

[0014] The present invention further provides a compound represented by the following formula (II): [ka]

[0015] [During the ceremony, X 1 represents a natural or unnatural amino acid, q represents an integer of 0 or 1; X 2 represents the natural amino acid I, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents the natural amino acid S, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R), X 6 represents the natural amino acid S, X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A), X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H), X 9represents the natural amino acid P or the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), X 10 represents the natural amino acid I, X 11 represents an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I, X 13 represents the natural amino acid P, s represents an integer of 0 or 1; X 14 represents a natural amino acid selected from the list consisting of D, Q and E, t represents an integer of 0 or 1; wherein the N-terminus of the peptide is unsubstituted, acetylated, or C1-C 20 -alkyl mono- or di-substituted, the C-terminus of the peptide is unsubstituted or amidated, and The peptide is cyclized and preferably has X 3 and X 11 is cyclized via a linkage connecting The present invention provides compounds comprising the peptide or derivatives, prodrugs, analogs, pharmaceutically acceptable salts, solvates or solvates of the salts of DETAILED DESCRIPTION OF THE INVENTION

[0016] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. Generally, the nomenclature used in connection with, and techniques of, chemistry, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0017] Throughout this specification, the term "comprise" or variations thereof, such as "comprises" or "comprising," are understood to mean the inclusion of a specified integer (or component) or group of integers (or components), but not the exclusion of other integers (or components) or groups of integers (or components). The singular forms "a," "an," and "the" include the plural unless the context clearly dictates otherwise. The terms "including" and "containing" are used to mean "including but not limited to," and these terms can be used interchangeably. In particular, the term "peptide-containing compound" refers to a compound that contains a defined peptide sequence and may optionally contain additional chemical groups or substituents covalently attached to the peptide, such as amino acids, fatty acids, or chemical groups that enhance the pharmacodynamic or pharmacokinetic properties of the peptide or any other chemical group. It should also be understood that the term "peptide-containing compound" expressly includes a defined peptide sequence without additional chemical groups or substituents covalently attached to the peptide.

[0018] As used herein, the following terms have the meanings ascribed to them unless otherwise specified: "Essentially consisting of" is understood to mean that the peptide is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the peptide to which it is being compared.

[0019] The terms "protein," "polypeptide," and "peptide" are used interchangeably to broadly refer to a sequence of two or more amino acids linked together, preferably by a peptide (amide) bond. A peptide (amide) bond is formed when the carboxyl group of one amino acid reacts with the amino group of another amino acid. It should be further understood that the terms "protein," "polypeptide," and "peptide" do not denote a specific length of a polymer of amino acids, nor are they intended to imply or distinguish whether a polypeptide is produced using recombinant technology, chemical synthesis, enzymatic synthesis, or naturally occurring. Furthermore, it should be understood that a peptide may contain one or more moieties that are not amino acids under the definition of this application. These moieties are preferably present at the N- and C-termini of the peptide.

[0020] As used herein, "amino acid" or "any amino acid" refers to an organic compound containing an amine (-NH) and a carboxyl (-COOH) functional group in the side chain, and refers to any and all amino acids, including naturally occurring amino acids (e.g., α-L-amino acids), unnatural amino acids, modified amino acids, and non-natural amino acids. "Natural amino acids" include those found in nature, such as the 23 amino acids that are linked into peptide chains to form the building blocks of many proteins. These are primarily L-stereoisomers, although D-amino acids occur in bacterial envelopes and some antibiotics. Table 2 lists the 20 naturally occurring proteinogenic amino acids in the standard genetic code. "Non-standard" natural amino acids are pyrrolysine (found in methanogens and other eukaryotes), selenocysteine ​​(present in many non-eukaryotes and most eukaryotes), and N-formylmethionine (encoded by the start codon AUG in bacteria, mitochondria, and chloroplasts).

[0021] "Unnatural" or "non-natural" amino acids are non-proteinogenic amino acids (i.e., amino acids that are not naturally encoded or found in the genetic code) that are either naturally occurring or chemically synthesized. Over 140 naturally occurring amino acids are known, with thousands of possible combinations. Examples of "unnatural" amino acids include the β-amino acids (β 3 and β 2 ), homoamino acids, proline and pyruvate derivatives, 3-substituted alanine derivatives, glycine derivatives, ring-substituted phenylalanine and tyrosine derivatives, linear nucleus amino acids, diamino acids, D-amino acids, and N-methylamino acids. Non-natural or unnatural amino acids also include modified amino acids. "Modified" amino acids include amino acids (e.g., natural amino acids) that have been chemically modified to include a group, groups, or chemical moiety that does not naturally occur in amino acids. In accordance with the present invention, preferred non-natural amino acids are listed in Table 1. While Table 1 presents the non-natural amino acids as D- and / or L-stereoisomers, preferred non-natural amino acids in accordance with the present invention are both the D- and L-stereoisomers of the non-natural amino acids listed in Table 1.

[0022] Table 1: Preferred Unnatural Amino Acids [Table 1] TIFF0007667091000006.tif255166TIFF0007667091000007.tif254169TIFF00076670910 00008.tif255169TIFF0007667091000009.tif254168TIFF0007667091000010.tif136168

[0023] More preferred unnatural amino acids are N-methyl-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, L-3-bromophenylalanine ((3-bromo)F), L-N,N-dimethylalanine ((N,N-diMe)A), N,N-dimethylglycine ((N,N-diMe)G), N-phenylglycine ((N-Ph)G), (R)-piperidine-3-carboxylic acid, (S)-piperidine-3-carboxylic acid, L-tert-butyl Alanine ((tBu)A), L-2-pyridylalanine (2-Pal), L-3-pyridylalanine (3-Pal), L-4-pyridylalanine (4-Pal), 3-(aminomethyl)benzoic acid, 3-amino-2,2-dimethylpropionic acid, 3-amino-3-methylbutyric acid, 4-(aminomethyl)benzoic acid, L-2-aminobutyric acid (Abu), 1-aminocyclobutane-1-carboxylic acid (ACBA), 6-aminohexanoic acid (Ahx), 2-aminoisobutyric acid (Aib), L -2-Thienylalanine (beta-2-thienylalanine), beta-alanine (beta-A), beta-proline (beta-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), gamma-aminobutyric acid (gamma-Abu), L-3-methylhistidine (3-Me)H), L-dihydroorotic acid (Hoo), L-norleucine (Nle), N-methyl-L-proline ((N-Me)P), L-nor Selected from the list consisting of valine (Nva), L-ornithine (Orn), L-pipecolic acid (Pip), (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid; 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), L-N-methylcysteine ​​((N-Me)C), N(5)-methyl-L-arginine ((Me)R), L-penicillamine (Pen) and tranexamic acid (tranexamic acid).

[0024] The most preferred unnatural amino acids are selected from the list consisting of N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva), L-ornithine (Orn), N(5)-methyl-L-arginine ((Me)R), L-tert-butylalanine ((tBu)A), 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), LN-methylcysteine ​​((N-Me)C), and L-penicillamine (Pen).

[0025] Furthermore, it should be understood that the peptides according to the invention may contain one or more chemical groups which are not amino acids under the definition of the present invention. These chemical groups may be present at the N- and / or C-terminus of the peptide and may be represented by the formula X 0 and X 15 It should be understood that all amino acids and chemical groups of the peptides of the present invention are linked via peptide (amide) bonds. Generally, peptides are formed by linking the α-amino and carboxy groups of α-amino acids, and the α-amino acids are linked via α-peptide bonds. According to the present invention, peptide bonds can be formed through any carboxyl and amino groups present in each natural or unnatural amino acid. For example, α-amino acids containing a second amino group in addition to the α-amino group (e.g., L-lysine) or α-amino acids containing a second carboxy group in addition to the α-carboxy group (e.g., L-aspartic acid and L-glutamic acid) can be linked via the additional amino or carboxy group.

[0026] As will be understood by those skilled in the art, the peptide sequences disclosed herein represent sequences of amino acids linked via α-peptide bonds. Amino acids linked via peptide bonds other than α-peptide bonds are marked with a "*". The "*" may be on either the left or right side of an amino acid to indicate that an additional amino group or an additional carboxy group of that amino acid is used in a peptide bond to an adjacent amino acid (e.g., (*L), (e*), (*Dap), etc.).

[0027] As will be understood by those skilled in the art, peptide sequences disclosed herein proceed from left to right, with the left-most end of the sequence designated as the "N-terminus" ("amino terminus," "N-terminal end") of the peptide and the right-most end of the sequence designated as the "C-terminus" ("carboxy terminus," "C-terminal end") of the peptide. The term N-terminus (amino terminus, N-terminal) applies regardless of whether the peptide actually contains an amino group at the N-terminus. The term C-terminus (carboxy terminus, C-terminal) applies regardless of whether the peptide actually contains a carboxy group at the C-terminus. The term "terminal amino group" refers to any amino group present at the N-terminus, and the term "terminal carboxyl group" refers to any carboxyl group present at the C-terminus.

[0028] According to the present invention, when p represents 1, the N-terminus is X 0 Alternatively, when q is at least 1 and p is 0, the N-terminus can be formed by X 1 Alternatively, when r is 1 and p and q are both 0, the N-terminus can be formed by X 2 When p, q and r all represent 0, the N-terminus can be X in the case of a linear peptide. 3 It is formed by X 3 and X 11 and p, q and r all represent 0, the peptide does not contain an N-terminus.

[0029] According to the present invention, when u represents 1, the C-terminus is X 15 Alternatively, when t is an integer of at least 1 and u is 0, the C-terminus can be formed by X 14 Alternatively, when s represents 1 and t and u both represent 0, the C-terminus can be formed by X 13 When s, t, and u are all 0, the C-terminus can be formed by X 12 is formed by

[0030] In the present invention, the names of natural and unnatural aminoacyl residues used herein preferably follow the naming rules suggested by the IUPAC Commission on Organic Chemistry Nomenclature and the IUPAC-IUB Commission on Biochemical Nomenclature as set out in Nomenclature of α-Amino Acids (Recommendations, 1974), Biochemistry, 14(2), (1975).

[0031] As used herein, naturally occurring proteinogenic amino acids are usually designated by their conventional one-letter abbreviations. Alternatively, they may also be referred to by their three-letter abbreviations (e.g., particularly in sequence listings) or by their full names as shown in Table 2 below: Table 2: Standard abbreviations for natural amino acids [Table 2]

[0032] For non-proteinogenic or non-naturally occurring amino acids, unless they are referred to by their full name (e.g., ornithine), the frequently used 3- to 6-letter codes, including abbreviations as shown in the following list of abbreviations (Table 3), are used for those residues.

[0033] As used herein, the term "L-amino acid" refers to the "L" isomeric form of an amino acid, and conversely, the term "D-amino acid" refers to the "D" isomeric form of an amino acid. Furthermore, it is conventional to designate L-amino acids with uppercase letters, such as Ala / A, Arg / R, and D-amino acids with lowercase letters, such as ala / a, arg / r.

[0034] The formats shown in Table 2 above, i.e., Ala, Arg, Asn, etc., and the three-letter codes generally used herein, are intended to generally include D- and L-forms as well as homo- and nor-forms, unless expressly indicated otherwise. The prefix "nor" refers to a structural analogue that can be derived from a parent compound by removing one carbon atom and its associated hydrogen atoms. The prefix "homo" indicates the next higher member in a homologous series. Reference to a specific isomeric form is indicated by the capitalized L- or D- prefixes above (e.g., D-Arg, L-Arg, etc.). Accordingly, specific reference to a homo or nor form is explicitly indicated by the respective prefix (e.g., homo-Arg, homo-R, nor-Arg, nor-R, homo-Cys, homo-C, etc.).

[0035] The term "C1-C6-alkyl" means a linear or branched saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5 or 6 carbon atoms, such as a methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neo-pentyl, 1,1-dimethylpropyl, hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1-ethylbutyl, 2-ethylbutyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2,3-dimethylbutyl, 1,2-dimethylbutyl or 1,3-dimethylbutyl group, or an isomer thereof. In particular, the radicals have 1, 2, 3 or 4 carbon atoms ("C1-C4-alkyl"), such as methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl or tert-butyl radicals, more particularly 1, 2 or 3 carbon atoms ("C1-C3-alkyl"), such as methyl, ethyl, n-propyl or isopropyl radicals. Particularly preferred are methyl, ethyl and n-propyl. Most preferred is methyl.

[0036] The term “C1-C 20"-Alkyl" means a linear or branched saturated monovalent hydrocarbon radical having 1 to 20 carbon atoms, e.g., methyl, ethyl, propyl, isopropyl, butyl, sec-butyl isobutyl, tert-butyl or pentyl, isopentyl, hexyl, isohexyl, heptyl, isoheptyl, octyl and isooctyl, nonyl, decyl, dodecyl or eicosyl.

[0037] The term "C1-C4-alkylene" means a linear or branched hydrocarbon bridge having 1 to 4 carbon atoms, such as methylene, ethylene, propylene, (α-methylethylene, β-methylethylene, α-ethylethylene, β-ethylethylene, butylene, α-methylpropylene, β-methylpropylene and γ-methylpropylene).

[0038] The term "C1-C6-alkylene" means a linear or branched hydrocarbon bridge having from 1 to 6 carbon atoms, such as methylene, ethylene, propylene, (α-methylethylene, β-methylethylene, α-ethylethylene, β-ethylethylene, butylene, α-methylpropylene, β-methylpropylene, γ-methylpropylene, α-ethylpropylene, β-ethylpropylene, γ-ethylpropylene, pentylene and hexylene.

[0039] The term "C3-C8-cycloalkyl" refers to a saturated hydrocarbon ring containing 3, 4, 5, 6, 7 or 8 carbon atoms. The C3-C8-cycloalkyl group is, for example, a monocyclic hydrocarbon ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl group, a bicyclic hydrocarbon ring, such as bicyclo[4.2.0]octyl or octahydropentalenyl, or a bridged or caged saturated ring group, such as norborane or adamantane, and cubane.

[0040] The term "C3-C7-heterocycloalkyl" denotes a saturated heterocycle having 4, 5, 6 or 7 rings containing one or two identical or different ring heteroatoms from the N, O and S series, said heterocycloalkyl group being bonded to the rest of the molecule via any one of the carbon atoms or, if present, via the nitrogen atom. The C3-C7-heterocycloalkyl group may be, for example, but not limited to, a 4-membered ring, such as azetidinyl, oxetanyl or thietanyl; or for example, a 5-membered ring, such as tetrahydrofuranyl, 1,3-dioxolanyl, thiolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, 1,1-dioxidethiolanyl, 1,2-oxazolidinyl, 1,3-oxazolidinyl or 1,3-thiazolidinyl; or for example, a 6-membered ring, such as tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, hexahydropyrimidinyl, 1,3-dioxanyl, 1,4-dioxanyl or 1,2-oxazinanyl; or for example, a 7-membered ring, such as azepanyl, 1,4-diazepanyl or 1,4-oxazepanyl.

[0041] The term "aryl" means an unsaturated or partially unsaturated ring having 6 to 10 carbon atoms. Preferred aryl groups are phenyl and naphthyl.

[0042] The term "heteroaryl" means a monovalent, monocyclic, bicyclic, or tricyclic aromatic ring having 5, 6, 8, 9, 10, 11, 12, 13, or 14 ring atoms (a "5- to 14-membered heteroaryl" group), in particular 5, 6, 9, or 10 ring atoms, which contains at least one ring heteroatom from the series: N, O, and / or S, and optionally 1, 2, or 3 further ring heteroatoms, bonded via a ring carbon atom or, optionally, via a ring nitrogen atom (where allowed by valence). The heteroaryl group may be, for example, a 5-membered heteroaryl group such as thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl or tetrazolyl; or, for example, a 6-membered heteroaryl group such as pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl; or, for example, a tricyclic heteroaryl group such as carbazolyl, acridinyl or or phenazinyl; or for example, a 9-membered heteroaryl group such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzothiazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl, indolizinyl or purinyl; or for example, a 10-membered heteroaryl group such as quinolinyl, quinazolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl or pteridinyl.

[0043] In general, unless otherwise specified, a heteroaryl or heteroarylene group includes all possible isomeric forms thereof, such as tautomers and positional isomers with respect to the point of attachment to the remainder of the molecule. Thus, in some illustrative, non-limiting examples, the term pyridinyl includes pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl, or the term thienyl includes thien-2-yl and thien-3-yl.

[0044] Disclosed herein are sequences incorporating either an "-OH" or "-NH" moiety at the carboxy terminus (C-terminus) of the sequence. The "-OH" or "-NH" moiety at the C-terminus of the sequence indicates a hydroxy group or an amino group, respectively, corresponding to the presence of a carboxy group or an amide (-(C=O)-NH) group at the C-terminus. In each of the sequences of the present invention, the C-terminal "-OH" moiety may be replaced with a C-terminal "-NH" moiety, also referred to herein as an "amidated C-terminus," and vice versa. However, among these alternatives, the C-terminal "-OH" moiety is preferred.

[0045] The term "acetylated" (also abbreviated as "Ac") refers to acetyl protection of the N-terminal moiety by acetylation of the N-terminus of the peptide (the N-terminus of the peptide is acetylated).

[0046] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or solvate of a salt, which may be isolated and / or purified, comprising, consisting essentially of or consisting of formula (I), wherein: X 0are (1S,2S,4S)-bicyclo[2.2.1]hept-5-en-2-ylacetic acid, (2,4-dioxoimidazolidin-1-yl)acetic acid, 2-(thiomorpholine)acetic acid ((2-thiomorpholine)acetyl), 2-(N-isopropyl-N-methylamino)acetic acid, (S)-3-methylvaleric acid ((S)-3-methylpentanoic acid), 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetic acid (2-(cyclohexylamino)acetyl), 2-(diethylamino)acetic acid (2-(diethylamino)acetyl), 2-(morpholine)acetic acid (2-(morpholine)acetyl), 2-(N-methyl-N-cyclopropylamino)acetic acid (2-(N-methyl) N-cyclopropylamino)acetyl), 2-(piperidine)acetic acid (2-(piperidine)acetyl), 2-(pyrrolidine)acetic acid (2-(pyrrolidine)acetyl), 2-hydroxyacetic acid (2-hydroxyacetyl), 2-hydroxyisobutyric acid (2-hydroxyisobutyric), 3-(aminomethyl)benzoic acid, 3-methoxypropionic acid, 4-(aminomethyl)benzoic acid, 4-methylpentanoic acid (4-methylvaleric), 5-chlorothiophene-carboxylic acid, 1-(aminomethyl)-cyclopropyl-1-carboxylic acid (ACMP), adipic acid, (S)-azetidine-2-carboxylic acid, benzoic acid (benzoic), 4-(3,5-dimethyl-1,2-Oxazol-4-yl)-L-phenylalanine, cyclobutanecarboxylic acid, 2-(cyclobutyl)acetic acid, cyclobutylacetic acid, cyclohexylacetic acid, cyclohexanecarboxylic acid, cyclopentanecarboxylic acid, cyclopentylacetic acid, cyclopropanecarboxylic acid, cyclopropanecarboxylic acid, a chemical group selected from the list consisting of: propylacetic acid, D-(+)-biotin, fumaric acid, 3-phenylpropanoic acid (hydrocinnamic), isobutyric acid, isovaleric acid (isovaleric), L-(+)-lactic acid (lactic), phenylacetic acid, piperidin-4-ylacetic acid, pivalic acid (pivalic), suberic acid, tert-butylacetic acid, tetrahydropyranyl-4-acetic acid, tetrahydro-2H-pyran-3-ylacetic acid, and trans-2-(3-((t-butoxy)carbonylamino)cyclohexyl)acetic acid), p represents an integer of 0 or 1; X 1represents a natural amino acid selected from the list consisting of A, F, G, H, I, K, L, P, R, S, T, V, W and Y, or represents N-methyl-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, L-3-bromophenylalanine ((3-bromo)F), LN,N-dimethylalanine ((N,N-diMe)A), N,N-dimethylglycine ((N,N-diMe)G), N-phenylglycine ((N-Ph)G), (R)-piperidine-3-carboxylic acid, (S)-piperidine-3-carboxylic acid, Peridine-3-carboxylic acid, L-tert-butylalanine ((tBu)A), L-2-pyridylalanine (2-Pal), L-3-pyridylalanine (3-Pal), L-4-pyridylalanine (4-Pal), 3-(aminomethyl)benzoic acid, 3-amino-2,2-dimethylpropionic acid, 3-amino-3-methylbutyric acid, 4-(aminomethyl)benzoic acid, L-2-aminobutyric acid (Abu), 1-aminocyclobutane- 1-Carboxylic acid (ACBA), 6-aminohexanoic acid (Ahx), 2-aminoisobutyric acid (Aib), L-2-thienylalanine (beta-2-thienylalanine), beta-alanine (beta-A), beta-proline (beta-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), gamma-aminobutyric acid (gamma-Abu), L-3-methylhistidine represents a natural amino acid selected from the list consisting of thiazol-3-yl (3-Me)H), L-dihydroorotic acid (Hoo), L-norleucine (Nle), N-methyl-L-proline ((N-Me))P), L-norvaline (Nva), L-ornithine (Orn), L-pipecolic acid (Pip), (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid, and tranexamic acid; On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, q represents an integer from 0 to 5; X 2represents the natural amino acid I or an unnatural amino acid selected from the list consisting of LN-methylisoleucine ((N-Me)I), allo-L-isoleucine (allo-I), L-cyclobutylalanine (Cba), L-norvaline (Nva), L-2-aminobutyric acid (Abu), (2S,3S)-2-[(3R)-3-amino-2-oxopyrrolidin-1-yl]-3-methylpentanoic acid, (2S,3S)-2-[2-oxopiperazin-1-yl]-3-methylpentanoic acid, and (2S,3S)-2-[(3S)-2-oxopiperazin-1-yl]-3-methylpentanoic acid; On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents a natural amino acid selected from the list consisting of S, C, T, R and K, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R), X 6 represents the natural amino acid S or an unnatural amino acid selected from the list consisting of allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap), X 7represents a natural amino acid selected from the list consisting of L and N, or is a 2,5-difluoro-L-phenylalanine ((2,5-difluoro)F), L-2-bromophenylalanine ((2-bromo)F), 2-chloro-L-phenylalanine ((2-chloro)F), L-4-bromophenylalanine ((4-bromo)F), 4-fluoro-L-leucine ((4-fluoro)L), L-tert-butylalanine ((tBu)A), L-tert-butylglycine ((tBu)G), 5,5,5-trifluoro-L-leucine (((trifluoro)L), (S)-2-(amino)-1,6-hexyl represents an unnatural amino acid selected from the list consisting of 2-amino-7-(tert-butoxy)-7-oxoheptanoic acid, 2-amino-3-(1-methylcyclopropyl)propanoic acid, 2-amino-4-cyanobutyric acid (Cnba), ... X 8 represents the natural amino acid P or an unnatural amino acid selected from the list consisting of (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, trans-4-fluoroproline ((trans-4-fluoro)P), L-hydroxyproline (Hyp), (3S)-morpholine-3-carboxylic acid (morpholine-3-carboxylic acid), L-pipecolic acid (Pip), and (4aR,6aR,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid; X 9represents the natural amino acid P or 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S,4S)-4-trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid, (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid , (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-trans-3-hydroxyproline ((3S-OH)P, trans-4-fluoroproline ((trans-4-fluoro)P), L-hydroxyproline (Hyp), (2S,4S)-4-fluoroproline ((cis-4-fluoro)P), (3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1) and (3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2), X 10 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, 3-chlorophenylglycine ((3-chloro-Ph)G), (S)-2-amino-3-ethylpentanoic acid, allo-L-isoleucine (allo-I), L-cyclohexylglycine (Chg), L-cyclopentylglycine (Cpg), L-cyclobutylglycine, and L-norvaline (Nva); X 11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12represents the natural amino acid I or an unnatural amino acid selected from the list consisting of 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, (S)-2-amino-2-cyclobutylacetic acid (Cbg), allo-L-isoleucine (allo-I), L-phenylglycine (Phg), 2-methyl-D-alloisoleucine, and (2S,3S)-2-((amino)methyl)-3-methylpentanoic acid; And if u, t, and s are all 0, then X 12 the terminal carboxyl group of which is unsubstituted or amidated, X 13 represents a natural amino acid selected from the list consisting of P, A and D, or represents a non-natural amino acid selected from the list consisting of 2-methyl-L-proline (2-Me)P, N-methyl-glycine ((N-Me)G), trans-4-fluoroproline ((trans-4-fluoro)P), L-2-aminobutyric acid (Abu), 2-aminoisobutyric acid (Aib), hydroxyproline (Hyp) and 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic); On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, and if u and t are both 0 and s is different from 0, then X 13 the terminal carboxyl group of which is unsubstituted or amidated, s represents an integer from 0 to 3, X 14represents a natural amino acid selected from the list consisting of D, E, G, K, N, P and Q, or represents an unnatural amino acid selected from the list consisting of 3-carboxyphenylalanine ((3-carboxy)F), (2S)-2-amino-4-(benzylamino)-4-oxobutanecarboxylic acid ((N-benzyl)D), N-methyl-glycine ((N-Me)G), 6-aminohexanoic acid (Ahx), (2S)-pyrrolidin-2-ylacetic acid (beta-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic); and if u is 0 and t is different from 0, then X 14 the terminal carboxyl group of which is unsubstituted or amidated, On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, t represents an integer from 0 to 4, X 15 is a chemical group selected from the list consisting of (1R,3S)-3-(amino)cyclopentanecarboxylic acid, (1S,3R)-3-(amino)cyclopentanecarboxylic acid, (R)-4-amino-6-methylheptanoic acid, (S)-(1-piperidin-3-yl)-acetic acid, (S)-3-(1-pyrrolidin-2-yl)-propionic acid, (S)-3-(2H-tetrazol-5-yl)propanoic acid, (S)-pyrrolidine-3-carboxylic acid, 5-azaspiro[2.4]heptane-1-carboxylic acid and (2S)-3-(triazol-1-yl)-2-(amino)propanoic acid, u represents an integer of 0 or 1; However, the peptide includes at least one unnatural amino acid.

[0047] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, which may be isolated and / or purified, comprising, consisting essentially of or consisting of formula (I), wherein X 0is a chemical group selected from the list consisting of (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, (2-thiomorpholine)acetic acid, (N-isopropyl-N-methylamino)acetic acid, 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetic acid, 2-(diethylamino)acetic acid, 2-(morpholine)acetic acid, 2-(piperidine)acetic acid, 2-(pyrrolidine)acetic acid, 3-(aminomethyl)benzoic acid, 4-(aminomethyl)benzoic acid, 1-(aminomethyl)-cyclopropyl-1-carboxylic acid (ACMP), azetidine-2-carboxylic acid, benzoic acid, cyclobutylacetic acid, cyclopropylacetic acid, phenylacetic acid, piperidin-4-ylacetic acid, tetrahydro-2H-pyran-3-ylacetic acid, tranexamic acid and trans-2-(3-((t-butoxy)carbonylamino)cyclohexyl)acetic acid, p represents an integer of 0 or 1; X 1represents a natural amino acid selected from the list consisting of A, F, G, H, I, K, L, P, R, S, T, V, W and Y, or is selected from the group consisting of LN,N-dimethylalanine ((N,N-diMe)A), N-methyl-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-2-pyridylalanine (2-Pal), L-3-pyridylalanine (3-Pal), L-4-pyridylalanine (4-Pal), L-2-aminobutyric acid (Abu), 6-aminohexanoic acid (Ahx), 2-aminoisobutyric acid (Aib), 3-(aminomethyl)benzoic acid, 3-amino-2,2-dimethylpropionic acid, 3-amino-3-methylbutyric acid, 4-(aminomethyl)benzoic acid, beta-2-thienylalanine, (S)-pyrrolidine-2- carboxylic acid (beta-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), gamma-aminobutyric acid (gamma-Abu), L-3-methylhistidine (H(3-Me)), L-dihydroorotic acid (Hoo), L-norleucine (Nle), L-norvaline (Nva), L-ornithine (Orn), L-pipecolic acid (Pip), N-methyl-L-proline ((N-Me)P), tranexamic acid (tranexamic), (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, and (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid; On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, q represents an integer from 0 to 5; X 2 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of LN-methylisoleucine ((N-Me)I), L-cyclobutylalanine (Cba), L-norvaline (Nva), and L-2-aminobutyric acid (Abu); On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, r represents an integer of 0 or 1; X3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents a natural amino acid selected from the list consisting of S and T, X 5 represents the natural amino acid R, X 6 represents the natural amino acid S or the unnatural amino acid allo-L-threonine (allo-T), X 7 represents a natural amino acid selected from the list consisting of N and L, or represents a natural amino acid selected from the list consisting of 2,5-difluoro-L-phenylalanine ((2,5-difluoro)F), L-2-bromophenylalanine ((2-bromo)F), 2-chloro-L-phenylalanine ((2-chloro)F), 4-fluoro-L-leucine ((4-fluoro)L), L-tert-butylalanine ((tBu)A), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propanoic acid and 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), 3-(trimethylsilyl)-L-alanine, X 8 represents the natural amino acid P or an unnatural amino acid selected from the list consisting of (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid and (4aR,6aR,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid; X 9represents the natural amino acid P or is an amino acid selected from the group consisting of 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), (2S,4S)-4-trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-azabicyclo[3.1.0]hexane-3 -carboxylic acid, (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid, rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1) and rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2), X 10 represents a natural amino acid I or an unnatural amino acid selected from the list consisting of (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, (S)-2-amino-3-ethyl-pentanoic acid, L-cyclohexylglycine (Chg) and L-cyclopentylglycine (Cpg); X 11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of allo-L-isoleucine (allo-I), (S)-2-amino-2-cyclobutylacetic acid (Cbg) and 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid; And if u, t, and s are all 0, then X 12 the terminal carboxyl group of which is unsubstituted or amidated, X 13represents a natural amino acid selected from the list consisting of P and D, or represents an unnatural amino acid selected from the list consisting of 2-aminoisobutyric acid (Aib), 2-methyl-L-proline (2-Me)P, and trans-4-fluoroproline ((trans-4-fluoro)P); On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, and if u and t are both 0 and s is different from 0, then X 13 the terminal carboxyl group of which is unsubstituted or amidated, s represents an integer from 0 to 3, X 14 represents a natural amino acid selected from the list consisting of D, Q, N, E and P, or represents a natural amino acid selected from the list consisting of 3-carboxyphenylalanine ((3-carboxy)F), N-methyl-glycine ((N-Me)G), (2S)-pyrrolidin-2-ylacetic acid (beta-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic), On the other hand, each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration, and if u is 0 and t is different from 0, then X 14 the terminal carboxyl group of which is unsubstituted or amidated, t represents an integer from 0 to 4, X 15 is a chemical group selected from the list consisting of (1R,3S)-3-(amino)cyclopentanecarboxylic acid, (1S,3R)-3-(amino)cyclopentanecarboxylic acid, (R)-4-amino-6-methylheptanoic acid, (S)-(1-piperidin-3-yl)-acetic acid, (S)-3-(1-pyrrolidin-2-yl)-propanoic acid, (S)-3-(2H-tetrazol-5-yl)propanoic acid, (S)-pyrrolidine-3-carboxylic acid, 5-azaspiro[2.4]heptane-1-carboxylic acid and (2S)-3-(triazol-1-yl)-2-(amino)propionic acid, u represents an integer of 0 or 1; However, the peptide includes at least one unnatural amino acid.

[0048] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or solvate of a salt, comprising, consisting essentially of or consisting of formula (I), which may be isolated and / or purified, wherein: p represents an integer equal to 0; X 1 represents a natural amino acid selected from the list consisting of A and G, or represents a non-natural amino acid selected from the list consisting of N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn); q represents an integer of 0 or 1; X 2 represents the natural amino acid I, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents the natural amino acid S, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R), X 6 represents the natural amino acid S, X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A), X 8 represents the natural amino acid P, X 9 represents the natural amino acid P or the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), X 10 represents the natural amino acid I, X11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of L-N-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I, And if u, t, and s are all 0, then X 12 the terminal carboxyl group of which is unsubstituted or amidated, X 13 represents the natural amino acid P, and if u and t are both 0 and s is different from 0, then X 13 the terminal carboxyl group of which is unsubstituted or amidated, s represents an integer of 0 or 1; X 14 represents a natural amino acid selected from the list consisting of D, Q and E, and if u is 0 and t is different from 0, then X 14 the terminal carboxyl group of which is unsubstituted or amidated, t represents an integer of 0 or 1; u represents the integer 0, However, the peptide contains at least one unnatural amino acid.

[0049] According to a further embodiment, the present invention provides a compound of formula (II): [ka]

[0050] [During the ceremony, X 1 represents a natural amino acid selected from the list consisting of A and G, or represents a non-natural amino acid selected from the list consisting of N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn); q represents an integer of 0 or 1; X2 represents the natural amino acid I, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents the natural amino acid S, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R), X 6 represents the natural amino acid S, X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A), X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H), X 9 represents the natural amino acid P or the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), X 10 represents the natural amino acid I, X 11 represents an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I, X 13 represents the natural amino acid P, s represents an integer of 0 or 1; X 14 represents a natural amino acid selected from the list consisting of D, Q and E, t represents an integer of 0 or 1; wherein the N-terminus of the peptide is unsubstituted, acetylated, or C1-C 20 - alkyl mono- or di-substituted, the C-terminus of the peptide is unsubstituted or amidated, and The peptide is X 3and X 11 is cyclized via a linkage connecting The present invention provides compounds comprising the peptide or derivatives, prodrugs, analogs, pharmaceutically acceptable salts, solvates or solvates of the salts of

[0051] According to a further embodiment, the present invention provides a peptide of formula (II) or a derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or compound containing solvate, wherein: X 1 represents a natural or unnatural amino acid, q represents an integer of 0 or 1; X 2 represents the natural amino acid I, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents the natural amino acid S, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R), X 6 represents the natural amino acid S, X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A), X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H), X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), X 10 represents the natural amino acid I, X 11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of L-N-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I, X 13 represents the natural amino acid P, s represents an integer of 0 or 1; X 14 represents a natural amino acid selected from the list consisting of D, Q and E, t represents an integer of 0 or 1; wherein the N-terminus of the peptide is unsubstituted, acetylated, or C1-C 20 -alkyl, and the C-terminus of the peptide is unsubstituted or amidated; and The peptide is X 3 and X 11 is cyclized via a linkage connecting

[0052] According to a further embodiment, the present invention provides a compound comprising a peptide of formula (II) or a derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or solvate of the salt, wherein: X 1 represents a natural amino acid selected from the list consisting of A and G, or represents a non-natural amino acid selected from the list consisting of N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn); q represents an integer of 0 or 1; X 2 represents the natural amino acid I, r represents an integer of 0 or 1; X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen), X 4 represents the natural amino acid S, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R), X 6 represents the natural amino acid S, X 7represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A), X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H), X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), X 10 represents the natural amino acid I, X 11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of L-N-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen); X 12 represents the natural amino acid I, X 13 represents the natural amino acid P, s represents an integer of 0 or 1; X 14 represents a natural amino acid selected from the list consisting of D, Q and E, t represents an integer of 0 or 1; wherein the N-terminus of the peptide is unsubstituted or acetylated; the C-terminus of the peptide is unsubstituted or amidated, and The peptide is X 3 and X 11 is cyclized via a linkage connecting

[0053] In particular, the present invention provides a compound containing a peptide of formula (I) or a compound containing a peptide of formula (II).

[0054] According to the present invention, the peptides of formula (I) and formula (II) contain the following X 1 ~X 14 and the peptide of formula (I) has a position X 0 and X 15 The definition of

[0055] According to the present invention, X 0 If present, may be a chemical group that is not an amino acid as defined herein.

[0056] According to a further embodiment of the present invention, X 0are 2-cyanobenzoic acid, (1S,2S,4S)-bicyclo[2.2.1]hept-5-en-2-ylacetic acid, (2,4-dioxoimidazolidin-1-yl)acetic acid, 2-(thiomorpholine)acetic acid ((2-thiomorpholine)acetyl), 2-(N-isopropyl-N-methylamino)acetic acid, (S)-3-methylvaleric acid ((S)-3-methylpentanoic acid), 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetic acid (2-(cyclohexylamino)acetyl), 2-(diethylamino)acetic acid (2-(diethylamino)acetyl), 2-(morpholine)acetic acid (2-(morpholine)acetyl), 2-(N-methyl-N-cyclopropylamino)acetic acid (2- (N-methyl-N-cyclopropylamino)acetyl), 2-(piperidine)acetic acid (2-(piperidine)acetyl), 2-(pyrrolidine)acetic acid (2-(pyrrolidine)acetyl), 2-hydroxyacetic acid (2-hydroxyacetyl), 2-hydroxyisobutyric acid (2-hydroxyisobutyric), 3-(aminomethyl)benzoic acid, 3-methoxypropionic acid, 4-(aminomethyl)benzoic acid, 4-methylpentanoic acid (4-methylvaleric), 5-chlorothiophene-carboxylic acid, 1-(aminomethyl)-cyclopropyl-1-carboxylic acid (ACMP), adipic acid, (S)-azetidine-2-carboxylic acid, benzoic acid, 4-(3,5-dimethyl-1,2-Oxazol-4-yl)-L-phenylalanine, cyclobutanecarboxylic acid, 2-(cyclobutyl)acetic acid, cyclobutylacetic acid, cyclohexylacetic acid, cyclohexanecarboxylic acid, cyclopentanecarboxylic acid, cyclopentylacetic acid, cyclopropanecarboxylic acid, cyclopropanecarboxylic acid, represents a chemical group selected from the list consisting of: propylacetic acid, D-(+)-biotin, fumaric acid, 3-phenylpropanoic acid (hydrocinnamic), isobutyric acid, isovaleric acid (isovaleric), L-(+)-lactic acid (lactic), phenylacetic acid, piperidin-4-ylacetic acid, pivalic acid (pivalic), suberic acid, tert-butylacetic acid, tetrahydropyranyl-4-acetic acid, tetrahydro-2H-pyran-3-ylacetic acid and trans-2-(3-((t-butoxy)carbonylamino)cyclohexyl)acetic acid).

[0057] According to a further embodiment of the present invention, X 0 are 2-cyanobenzoic acid, (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, (2-thiomorpholine)acetic acid, (N-isopropyl-N-methylamino)acetic acid, 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetic acid, 2-(diethylamino)acetic acid, 2-(morpholine)acetic acid, 2-(piperidine)acetic acid, 2-(pyrrolidine)acetic acid, 3-(aminomethyl)benzoic acid, 4-(amino represents a chemical group selected from the list consisting of trans-2-(3-((t-butoxy)carbonylamino)cyclohexyl)acetic acid, 1-(methyl)benzoic acid, 1-(aminomethyl)-cyclopropyl-1-carboxylic acid (ACMP), azetidine-2-carboxylic acid, benzoic acid, cyclobutylacetic acid, cyclopropylacetic acid, phenylacetic acid, piperidin-4-ylacetic acid, tetrahydro-2H-pyran-3-ylacetic acid, tranexamic acid, and trans-2-(3-((t-butoxy)carbonylamino)cyclohexyl)acetic acid.

[0058] According to a further embodiment of the present invention, X 0is a chemical group selected from an unnatural amino acid, 2-cyanobenzoic acid, a substituted benzoic acid, or a phenylacetic acid as defined above.

[0059] According to a further embodiment of the present invention, X 0 is Hoo, ODD or Ahx.

[0060] According to a further embodiment of the invention, p represents the integer 1.

[0061] According to a further embodiment of the invention, p represents the integer 0.

[0062] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 1 represents any natural amino acid or represents an unnatural amino acid, preferably selected from the list of unnatural amino acids listed in Table 1, while each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced by its stereoisomer in the D-configuration.

[0063] According to a further embodiment of the present invention, X 1represents a natural amino acid selected from the list consisting of A, F, G, H, I, K, L, P, R, S, T, V, W and Y, or represents N-methyl-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, L-3-bromophenylalanine ((3-bromo)F), L-N,N-dimethylalanine ((N,N-diMe)A), N,N-dimethylglycine ((N,N-diMe)G), N-phenylalanine ((3-bromo)F), L-N,N-dimethylalanine ((N,N-diMe)A), N,N-dimethylglycine ((N,N-diMe)G), N-phenylalanine ((3-bromo)F), L-N,N-dimethylalanine ((N,N-diMe)A), N-phenylalanine ((3-bromo)F), L-N,N-dimethylalanine ((N,N-diMe)A), N-phenylalanine ((3-bromo)F), L-N,N-dimethylalanine ((N,N-diMe)G ... L-glycine ((N-Ph)G), (R)-piperidine-3-carboxylic acid, (S)-piperidine-3-carboxylic acid, L-tert-butylalanine ((tBu)A), L-2-pyridylalanine (2-Pal), L-3-pyridylalanine (3-Pal), L-4-pyridylalanine (4-Pal), 3-(aminomethyl)benzoic acid, 3-amino-2,2-dimethylpropionic acid, 3-amino-3-methylbutyric acid, 4-(aminomethyl)benzoic acid, L-2-aminobutyric acid (Abu), 1-amino Cyclobutane-1-carboxylic acid (ACBA), 6-aminohexanoic acid (Ahx), 2-aminoisobutyric acid (Aib), L-2-thienylalanine (beta-2-thienylalanine), beta-alanine (beta-A), beta-proline (beta-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), gamma-aminobutyric acid (gamma-Abu), L-3-methylhistidine (3-MeH), L-dihydroorotic acid ( represents an unnatural amino acid selected from the list consisting of Hoo), L-norleucine (Nle), N-methyl-L-proline ((N-Me)P), L-norvaline (Nva), L-ornithine (Orn), L-pipecolic acid (Pip), (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid, and tranexamic acid (tranexamic acid), wherein each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration.

[0064] According to a further embodiment of the present invention, X 1represents a natural amino acid selected from the list consisting of A and G, or an unnatural amino acid selected from the list consisting of N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn), although the L-configuration natural amino acid and / or unnatural amino acid can be replaced with the D-configuration stereoisomer.

[0065] According to a further embodiment, X 1 represents a natural amino acid selected from the list consisting of A and G, or a non-natural amino acid selected from the list consisting of N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva), and L-ornithine (Orn).

[0066] According to a further embodiment, X 1 represents the natural amino acid A or the unnatural amino acid N-methyl-glycine ((N-Me)G).

[0067] According to a further embodiment of the present invention, when p is 0 and q is different from 0, X 1 The terminal amino group of is unsubstituted, acetylated or mono- or disubstituted with C1-C4-alkyl, preferably C1-C 20 - monosubstituted with alkyl.

[0068] According to a further embodiment of the present invention, X 1 is methylated.

[0069] According to a further embodiment of the present invention, X 1 is acetylated.

[0070] According to a further embodiment of the invention, q represents the integer 0.

[0071] According to a further embodiment of the invention, q represents the integer 1.

[0072] According to a further embodiment, there is provided a compound comprising a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 2 represents a natural amino acid selected from the list consisting of I, L, M, V and A, or is selected from the list consisting of LN-methylisoleucine ((N-Me)I), allo-L-isoleucine (allo-I), L-cyclobutylalanine (Cba), L-norvaline (Nva), L-2-aminobutyric acid (Abu), (2S,3S)-2-[(3R)-3-amino-2-oxopyrrolidin-1-yl]-3-methylpentanoic acid, (2S,3S)-2-[2-oxopyrrolidin-1-yl]-3-methylpentanoic acid, and (2S,3S)-2-[(3S)-2-oxopiperazin-1-yl]-3-methylpentanoic acid, (2S,3S)-2-[(3S)-2-oxopiperazin-1-yl]-3-methylpentanoic acid, L-methionine-L-sulfoxide, L-methionine-sulfone, and L-tert-butylglycine, wherein each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration.

[0073] According to a further embodiment of the present invention, X 2 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of LN-methylisoleucine ((N-Me)I), allo-L-isoleucine (allo-I), L-cyclobutylalanine (Cba), L-norvaline (Nva), L-2-aminobutyric acid (Abu), (2S,3S)-2-[(3R)-3-amino-2-oxopyrrolidin-1-yl]-3-methylpentanoic acid, (2S,3S)-2-[2-oxopiperazin-1-yl]-3-methylpentanoic acid, and (2S,3S)-2-[(3S)-2-oxopiperazin-1-yl]-3-methylpentanoic acid, while each natural and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration.

[0074] According to a further embodiment of the present invention, X 2 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of LN-methylisoleucine ((N-Me)I), L-cyclobutylalanine (Cba), L-norvaline (Nva) and L-2-aminobutyric acid (Abu), while each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced with its stereoisomer in the D-configuration.

[0075] According to a further embodiment of the present invention, X 2 represents natural amino acid I.

[0076] According to a further embodiment of the present invention, when p and q are both 0 and r is 1, X 2 The terminal amino group of is unsubstituted, acetylated, or C1-C 20 -alkyl, preferably mono- or di-substituted by C1-C4-alkyl.

[0077] According to a further embodiment of the invention, r represents the integer 0.

[0078] According to a further embodiment of the invention, r represents the integer 1.

[0079] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 3 represents the natural amino acid C or a non-natural amino acid selected from the list consisting of L-penicillamine (Pen) and LN-methylcysteine ​​((N-Me)C).

[0080] According to a further embodiment of the present invention, X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen).

[0081] According to a further embodiment of the present invention, X 3 represents the natural amino acid C.

[0082] According to a further embodiment of the present invention, when p, q and r are all 0, X 3 The terminal amino group of is unsubstituted, acetylated or mono- or disubstituted with C1-C4-alkyl, preferably C1-C 20 -substituted with alkyl.

[0083] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 4 represents a natural amino acid selected from the list consisting of S, C, T, R or K, or represents an unnatural amino acid selected from the list consisting of allo-L-threonine (allo-T), L-homoserine (hSer) and L-ornithine (Orn).

[0084] According to a further embodiment of the present invention, X 4 represents a natural amino acid selected from the list consisting of S, C, T, R and K.

[0085] According to a further embodiment of the present invention, X 4 represents a natural amino acid selected from the list consisting of S or T.

[0086] According to a further embodiment of the present invention, X 4 represents the natural amino acid S.

[0087] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R).

[0088] According to a further embodiment of the present invention, X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R).

[0089] According to a further embodiment of the present invention, X 5 represents the natural amino acid R.

[0090] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 6 represents a natural amino acid selected from the list consisting of S, C, or T, or represents an unnatural amino acid selected from the list consisting of allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap).

[0091] According to a further embodiment of the present invention, X 6 represents the natural amino acid S or an unnatural amino acid selected from the list consisting of allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap).

[0092] According to a further embodiment of the present invention, X 6 represents the natural amino acid S or the unnatural amino acid allo-L-threonine (allo-T).

[0093] According to a further embodiment of the present invention, X 6 represents the natural amino acid S.

[0094] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 7represents a natural amino acid selected from the list consisting of L, F or N, or is selected from the list consisting of 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), L-4-bromophenylalanine ((4-bromo)F), 2,5-difluoro-L-phenylalanine ((2,5-difluoro)F), L-tert-butylalanine ((tBu)A), 2-chloro-L-phenylalanine ((2-chloro)F), L-2-bromophenylalanine ((4-bromo)F), L-tert-butylglycine ((tBu)G), 3-(trimethylsilyl)-2-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propanoic acid, ... )-L-alanine, 2,5-difluoro-L-phenylalanine, 2-amino-7-(tert-butoxy)-7-oxoheptanoic acid, 5,5,5-trifluoro-L-leucine ((trifluoro)L), 2-methyl-L-phenylalanine ((2-Me)F), L-cyclobutylalanine (Cba), L-cyclopentylalanine (Cpa), L-cyclopropylmethylalanine, L-trifluoromethylalanine, L-difluoromethylalanine, 2 represents an unnatural amino acid selected from the list consisting of -fluoro-L-phenylalanine ((2-fluoro)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, (2S)-3-(3-cyanophenyl)-2-aminopropanoic acid, 2-amino-5,5,5-trifluoro-4-methyl-pentanoic acid, (2S)-2-amino-5-methyl-hexanoic acid and (2S)-3-(indol-4-yl)-2-(amino)propanoic acid.

[0095] According to a further embodiment of the present invention, X 7represents a natural amino acid selected from the list consisting of L and N, or 2,5-difluoro-L-phenylalanine ((2,5-difluoro)F), L-2-bromophenylalanine ((2-bromo)F), 2-chloro-L-phenylalanine ((2-chloro)F), L-4-bromophenylalanine ((4-bromo)F), 4-fluoro-L-leucine ((4-fluoro)L), L-tert-butylalanine ((tBu)A), L-tert-butylglycine ((tBu)G), 5,5,5-trifluoro-L-leucine ((trifluoro)L), (S)-2-(amino)-1,6-hexanedioic acid (AAD), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-2-amino-4-cyanobutyric acid (Cnb a) represents an unnatural amino acid selected from the list consisting of (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propanoic acid, (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), 3-(trimethylsilyl)-L-alanine, and 2-amino-7-(tert-butoxy)-7-oxoheptanoic acid.

[0096] According to a further embodiment of the present invention, X 7 represents a natural amino acid selected from the list consisting of N or L, or represents a natural amino acid selected from the list consisting of 2,5-difluoro-L-phenylalanine ((2,5-difluoro)F), L-2-bromophenylalanine ((2-bromo)F), 2-chloro-L-phenylalanine ((2-chloro)F), 4-fluoro-L-leucine ((4-fluoro)L), L-tert-butylalanine ((tBu)A), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propanoic acid, and 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), 3-(trimethylsilyl)-L-alanine.

[0097] According to a further embodiment of the present invention, X 7represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A).

[0098] According to a further embodiment of the present invention, X 7 represents the unnatural amino acid L-tert-butylalanine ((tBu)A).

[0099] According to a further embodiment of the present invention, X 7 represents the natural amino acid L.

[0100] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 8 represents the natural amino acid P, or L-proline (3,4-2H), (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, L-hydroxyproline (Hyp), (3S)-morpholine-3-carboxylic acid (morpholine-3-carboxylic acid), L-pipecolic acid (Pip), (4aR, represents a natural amino acid selected from the list consisting of (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, (1R,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid, trans-4-fluoroproline ((trans-4-fluoro)P), and (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid.

[0101] According to a further embodiment of the present invention, X 8represents the natural amino acid P or an unnatural amino acid selected from the list consisting of L-proline (3,4-2H), (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, trans-4-fluoroproline ((trans-4-fluoro)P), L-hydroxyproline (Hyp), (3S)-morpholine-3-carboxylic acid (morpholine-3-carboxylic acid), L-pipecolic acid (Pip), and (4aR,6aR,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid.

[0102] According to a further embodiment of the present invention, X 8 represents the natural amino acid P or an unnatural amino acid selected from the list consisting of L-proline (3,4-2H), (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid and (4aR,6aR,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid.

[0103] According to a further embodiment of the present invention, X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H).

[0104] According to a further embodiment of the present invention, X 8 represents the natural amino acid P.

[0105] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 9represents the natural amino acid P, or is an amino acid selected from the group consisting of 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), L-hydroxyproline (Hyp), (2S,4S)-4-trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,4S)-4-fluoroproline ((cis-4-fluoro)P), trans-4-fluoroproline ((trans-4-fluoro)P), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-trans-3-hydroxyproline ((3S-OH)P, (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-propanol, 2-( ... and (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid, trans-4-fluoroproline ((trans-4-fluoro)P), (2S,4S)-4-fluoroproline ((cis-4-fluoro)P), L-4,4-difluoroproline ((difluoro)P), rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1), and rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2).

[0106] According to a further embodiment of the present invention, X 9represents the natural amino acid P or 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S,4S)-4-trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid, (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid and (3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1), (3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2).

[0107] According to a further embodiment of the present invention, X 9 represents the natural amino acid P or is an amino acid selected from the group consisting of 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), (2S,4S)-4-trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-azabicyclo[3.1.0]hexane-3 -carboxylic acid, (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid, rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1) and rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2).

[0108] According to a further embodiment of the present invention, X 9 represents the natural amino acid P or the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic).

[0109] According to a further embodiment of the present invention, X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic).

[0110] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 10 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of L-cyclopentylglycine (Cpg), L-cyclohexylglycine (Chg), (S)-2-amino-3-ethyl-pentanoic acid, 3-chlorophenylglycine ((3-chloro-Ph)G), L-tert-butylglycine, allo-L-isoleucine (allo-I), L-cyclobutylglycine, L-norvaline (Nva), and (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid.

[0111] According to a further embodiment of the present invention, X 10 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, 3-chlorophenylglycine ((3-chloro-Ph)G), (S)-2-amino-3-ethyl-pentanoic acid, allo-L-isoleucine (allo-I), L-cyclohexylglycine (Chg), L-cyclopentylglycine (Cpg), L-cyclobutylglycine, and L-norvaline (Nva).

[0112] According to a further embodiment of the present invention, X 10 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, (S)-2-amino-3-ethyl-pentanoic acid, L-cyclohexylglycine (Chg) and L-cyclopentylglycine (Cpg).

[0113] According to a further embodiment of the present invention, X 10 represents natural amino acid I.

[0114] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0115] According to a further embodiment of the present invention, X 11 represents the natural amino acid C or an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0116] According to a further embodiment of the present invention, X 11 represents an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0117] According to a further embodiment of the present invention, X 11 represents the unnatural amino acid L-penicillamine (Pen).

[0118] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), and X 11 represents the unnatural amino acid LN-methylcysteine ​​((N-Me)C) or the unnatural amino acid L-penicillamine (Pen).

[0119] According to a further embodiment of the present invention, X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), and X 11 represents the unnatural amino acid L-penicillamine (Pen).

[0120] According to a further embodiment of the present invention, X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), and X 11 represents an unnatural amino acid selected from the list consisting of LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0121] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 12represents the natural amino acid I or an unnatural amino acid selected from the list consisting of allo-L-isoleucine (allo-I), (S)-2-amino-2-cyclobutylacetic acid (Cbg), (2S,3S)-2-((amino)methyl)-3-methylpentanoic acid, L-phenylglycine (Phg), 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, 2-methyl-D-alloisoleucine, L-norvaline (Nva), L-2-aminobutyric acid (Abu), L-tert-butylglycine, and aminoisobutyric acid (Aib).

[0122] According to a further embodiment of the present invention, X 12 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, (S)-2-amino-2-cyclobutylacetic acid (Cbg), allo-L-isoleucine (allo-I), L-phenylglycine (Phg), 2-methyl-D-alloisoleucine, and (2S,3S)-2-((amino)methyl)-3-methylpentanoic acid.

[0123] According to a further embodiment of the present invention, X 12 represents the natural amino acid I or an unnatural amino acid selected from the list consisting of allo-L-isoleucine (allo-I), (S)-2-amino-2-cyclobutylacetic acid (Cbg), and 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid.

[0124] According to a further embodiment of the present invention, X 12 represents natural amino acid I.

[0125] According to yet another embodiment of the present invention, when u, t, and s are all 0, then X 12 The terminal carboxyl group of is unsubstituted or amidated.

[0126] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 13 represents a natural amino acid selected from the list consisting of P, A, S, T, G, D, E, Q or N, or is selected from the group consisting of N-methyl-glycine ((N-Me)G), 5-azaspiro[2.4]heptane-6-carboxylic acid, L-2-aminobutyric acid (Abu), 2-aminoisobutyric acid (Aib), 2-methyl-L-proline (2-Me)P, hydroxyproline (Hyp), 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), trans-4-fluoroproline ((trans- (4-fluoro)P), (2S,4S)-4-fluoroproline ((cis-4-fluoro)P), L-4,4-difluoroproline ((difluoro)P), L-cyclopentylglycine (Cpg), (S)-2-amino-2-cyclobutylacetic acid (Cbg) and (2S)-pyrrolidin-2-ylacetic acid (beta-homo-P), wherein each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced by its stereoisomer in the D-configuration.

[0127] According to a further embodiment of the present invention, X 13 represents a natural amino acid selected from the list consisting of P, A and D, or an unnatural amino acid selected from the list consisting of 2-methyl-L-proline (2-Me)P, N-methyl-glycine ((N-Me)G), trans-4-fluoroproline ((trans-4-fluoro)P), L-2-aminobutyric acid (Abu), 2-aminoisobutyric acid (Aib), hydroxyproline (Hyp) and 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), although each natural and / or unnatural amino acid in the L-configuration can be replaced by its stereoisomer in the D-configuration.

[0128] According to a further embodiment of the present invention, X 13 represents a natural amino acid selected from the list consisting of P or D, or an unnatural amino acid selected from the list consisting of 2-aminoisobutyric acid (Aib), 2-methyl-L-proline (2-Me)P, and trans-4-fluoroproline ((trans-4-fluoro)P), although each natural and / or unnatural amino acid in the L-configuration can be replaced by its stereoisomer in the D-configuration.

[0129] According to a further embodiment of the present invention, X 13 represents the natural amino acid P.

[0130] According to a further embodiment of the invention, s represents the integer 0.

[0131] According to a further embodiment of the invention, s represents the integer 1.

[0132] According to yet another embodiment of the present invention, if u and t are both 0 and s is different from 0, then X 13 The terminal carboxyl group of is unsubstituted or amidated.

[0133] According to a further embodiment of the present invention, when t is 0 and s is 1, X 13 The terminal carboxyl group of is unsubstituted or amidated.

[0134] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein X 14 represents any natural or unnatural amino acid, where any natural and / or unnatural amino acid may be in the D- or L-configuration.

[0135] According to a further embodiment of the present invention, X 14 represents a natural amino acid selected from the list consisting of D, E, G, K, N, P and Q, or an unnatural amino acid selected from the list consisting of 3-carboxyphenylalanine ((3-carboxy)F), (2S)-2-amino-4-(benzylamino)-4-oxobutanecarboxylic acid ((N-benzyl)D), N-methyl-glycine ((N-Me)G), 6-aminohexanoic acid (Ahx), (2S)-pyrrolidin-2-ylacetic acid (beta-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic), although each natural amino acid and / or unnatural amino acid in the L-configuration can be replaced by its stereoisomer in the D-configuration.

[0136] According to a further embodiment of the present invention, X 14 represents a natural amino acid selected from the list consisting of D, Q, N, E and P, or an unnatural amino acid selected from the list consisting of 3-carboxyphenylalanine ((3-carboxy)F), N-methyl-glycine ((N-Me)G), (2S)-pyrrolidin-2-ylacetic acid (beta-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic), although each natural and / or unnatural amino acid in the L-configuration may be replaced by its stereoisomer in the D-configuration.

[0137] According to a further embodiment of the present invention, X 14 represents a natural amino acid selected from the list consisting of D, Q or I.

[0138] According to a further embodiment of the present invention, X 14 represents the natural amino acid D.

[0139] According to a further embodiment of the present invention, X 14 The C-terminal "-OH" moiety in is replaced with a C-terminal "-NH2" moiety.

[0140] According to a further embodiment of the invention, q represents the integer 0.

[0141] According to a further embodiment of the invention, q represents the integer 1.

[0142] According to a further embodiment of the invention, when s and t represent the integer 1, X 14 The terminal carboxyl group of is unsubstituted or amidated.

[0143] According to a further embodiment of the invention, when s and t represent integers of 1, X 14 The terminal carboxyl group of is not substituted.

[0144] According to the present invention, X 15 When present, may be a chemical group that is not an amino acid.

[0145] According to a further embodiment of the present invention, X 15 is a chemical group selected from the list consisting of -NH, -NH-C-C-alkyl, (1R,3S)-3-(amino)cyclopentanecarboxylic acid, (1S,3R)-3-(amino)cyclopentanecarboxylic acid, (R)-4-amino-6-methylheptanoic acid, (S)-(1-piperidin-3-yl)-acetic acid, (S)-3-(1-pyrrolidin-2-yl)-propionic acid, (S)-3-(2H-tetrazol-5-yl)propanoic acid, (S)-pyrrolidine-3-carboxylic acid, 5-azaspiro[2.4]heptane-1-carboxylic acid and (2S)-3-(triazol-1-yl)-2-(amino)propanoic acid.

[0146] According to a further embodiment of the present invention, X 15 is a chemical group selected from -NH2 or -NH-C1-C6-alkyl, preferably -NH2.

[0147] According to a further embodiment of the invention, q represents the integer 0.

[0148] According to a further embodiment of the invention, q represents the integer 1.

[0149] According to a further embodiment of the invention, the N-terminus of the peptide is unsubstituted, acetylated or C1-C 20 - mono- or di-substituted by alkyl;

[0150] According to a further embodiment of the invention, the N-terminus of the peptide is unsubstituted, acetylated or mono- or di-substituted with C1-C4-alkyl.

[0151] According to a further embodiment of the invention, the N-terminus of the peptide is unsubstituted or acetylated.

[0152] According to a further embodiment of the invention, the C-terminus is unsubstituted or amidated.

[0153] According to a further embodiment of the invention, the N- and C-termini are not substituted.

[0154] According to further embodiments, the present invention provides compounds containing peptides or derivatives, prodrugs, analogs, pharmaceutically acceptable salts, solvates or solvates of salts comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, selected from the group consisting of Examples 29, 44, 45, 67, 75, 109, 166, 185, 443 and 466, or selected from the group consisting of Examples 29, 44, 45, 67, 75, 109, 166, 185, 443, 466 and 499.

[0155] The peptides of formula (I) or (II) can be linear or cyclized. As used herein, the term "cyclized" refers to a reaction in which a portion of a polypeptide molecule (e.g., Cys, (N-Me)Cys, or Pen) is bonded to another portion of the polypeptide molecule (e.g., another Cys, (N-Me)Cys, or Pen) to form a closed ring, such as by forming a disulfide bridge (-SS-) or a carbon-sulfur bond (e.g., (-CH2-S-) or (-(CH2)2-S-)), a sulfur-carbon bond (e.g., (-S-CH2-) or (-S-(CH2)2-)), a carbon-sulfur-carbon bond (-CH2-S-CH2-) or other similar bond such as a carbon-carbon bond (e.g., (-CH2-CH2-)). According to the present invention, X of a peptide 3 Place and X 11 A linkage between the positions is preferred. In the sequences of peptides according to the invention, an amino acid followed by a "+" indicates a cyclic peptide, in which the amino acid is linked to another amino acid followed by a "+" by a disulfide bridge forming a closed ring. Alternatively, two linked chemical groups of a polypeptide molecule can be indicated by a bond line.

[0156] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of the salt comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein said peptide is cyclized.

[0157] According to a further embodiment of the present invention, the peptide of formula (I) or formula (II) comprises X 3 and X 11 is cyclized via a linkage joining

[0158] According to a further embodiment of the present invention, the peptide of formula (I) or formula (II) comprises X 3 and X 11 wherein X is cyclized via a linkage connecting 3 and X 11are attached via a linker of the formula -SS-, -CH2-S-, -S-CH2-, -CH2-CH2-, -S-(CH2)2-, -(CH2)2-S- or -CH2-S-CH2-.

[0159] According to a further embodiment of the present invention, the peptide of formula (I) or formula (II) comprises X 3 and X 11 wherein X is cyclized via a linkage connecting 3 and X 11 are linked via a disulfide bond of the formula -SS-.

[0160] The peptides of the present invention can be substituted with a suitable water-soluble polymer characterized by the repeating unit, which may be selected from the group consisting of polyalkyloxypolymers, hyaluronic acid and its derivatives, polyvinyl alcohol, polyoxazoline, polyanhydride, poly(orthoester), polycarbonate, polyurethane, polyacrylic acid, polyacrylamide, polyacrylate, polymethacrylate, polyorganophosphazene, polysiloxane, polyvinylpyrrolidone, polycyanoacrylate, and polyester.

[0161] The peptides of the present invention can be substituted with at least one polyethylene group (PEG group). At least one PEG group is preferably attached to the N-terminus and / or C-terminus. The PEG group can be attached to any suitable functional group of a chemical group and / or amino acid of the peptide (e.g., hydroxyl group, carboxyl group, amino group, thiol group, preferably amino group or carboxy group). Preferably, the peptides according to the present invention contain one PEG group attached to the N-terminus or one PEG group attached to the C-terminus. More preferably, one PEG group is attached to the N-terminus or C-terminus via an amide bond.

[0162] PEGylation of peptides can enhance their solubility, reduce immunogenicity, improve stability, and / or extend half-life by reducing renal clearance, a concept that has been known since the early 1980s (Caliceti P., Veronese FM, Adv. Drug Deliv. Rev. 2003, 55, 1261-1277). While this has been used successfully for some drugs, in many instances PEGylation reduces the efficacy of the drug substance to the extent that this concept is no longer appropriate (T. Peleg-Shulman et al., J. Med. Chem., 2004, 47, 4897-4904).

[0163] The PEG group according to the present invention is any group containing at least two ethylene oxide units to form an ethylene oxide oligomer or polymer. The PEG group is covalently attached to the peptide of the present invention (PEGylation), and is therefore called a PEGylated peptide. In general, this type of modification to molecules is well known in the art.

[0164] The PEG group can consist of an interconnecting portion, a polymer portion, and an end group. The interconnecting portion is C1-C 20 The alkyl may be composed of a heteroatom or an -O-, -S-, N(R X ), C(O), C(O)R X , C(O)N(R X ), N(R X )C(O), one or more C3-C8-cycloalkyl, C3-C7-heterocycloalkyl, aryl or heteroaryl, optionally interrupted or terminated by functional groups selected from the group consisting of: X is hydrogen or C1-C6-alkyl, which may be interrupted or terminated by one or more of the above atoms or groups, which further have hydrogen as a terminal atom. The polymeric portion consists of at least two ethylene oxide units, which are terminated by one or more heteroatoms or -O-, -S-, N(R X ), C(O), C(O)R X, C(O)N(R X ), N(R X )C(O), C1-C6-alkyl, C3-C8-cycloalkyl, C3-C7-heterocycloalkyl, aryl or heteroaryl, optionally interrupted or terminated by a functional group selected from the group consisting of R X is hydrogen or C1-C6-alkyl.

[0165] The terminal groups are heteroatoms or OH, SH, N(R X )2, C(O)R X ;COR X , COOR X , C(O)N(R X ), N(R X )C(O)NH2, where R X is hydrogen or C1-C6-alkyl. Preferred end groups are COOH and C(O)NH2.

[0166] Examples of PEG spacers according to the present invention include PEG1 (10 atoms) having 1 ethylene glycol unit (n=1; 10 atoms), PEG2 (13 atoms) having 2 ethylene glycol units (n=2; 13 atoms), PEG3 (16 atoms) having 3 ethylene glycol units (n=3; 16 atoms), PEG4 (19 atoms) having 4 ethylene glycol units (n=4; 19 atoms), PEG5 (22 atoms) having 5 ethylene glycol units (n=5; 22 atoms), PEG6 (25 atoms) having 6 ethylene glycol units (n=6; 25 atoms), etc.

[0167] It should be noted that PEG groups as defined herein may be designated by different names by commercial suppliers, and this does not exclude such identical compounds having different names from the present invention.

[0168] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of the salt, comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, wherein said peptide is substituted with at least one PEG group.

[0169] According to a further embodiment of the invention, the peptide is substituted by one PEG group.

[0170] According to a further embodiment of the invention, the peptide is substituted by one PEG group, wherein the PEG group is attached to the C-terminus, preferably via an amide bond.

[0171] According to a further embodiment of the invention, the peptide is substituted by at least one PEG group, at least one of which is of formula (IIIa) [ka]

[0172] [During the ceremony, * indicates a bond to a nitrogen or oxygen atom; D is C1-C4-alkylene, Y is selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, carboxamido, or amino; n represents an integer of 2 to 15. or a group of formula (IIIb) [ka]

[0173] [During the ceremony, * indicates a bond to a carbonyl group, D 1 is C1-C4-alkylene, Y 1is selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, carboxamido or amino; m represents an integer of 2 to 15. Is it a group of Or TTDS.

[0174] According to a further aspect of the invention, the peptide is substituted with at least one PEG group, with at least one or more PEG groups selected from the list consisting of PEG1 (10 atoms), PEG2 (13 atoms), PEG3 (16 atoms), PEG4 (19 atoms), PEG4-CH2CO2H (15 atoms), PEG5 (22 atoms), PEG5-CH2CO2H (18 atoms), PEG7 (25 atoms), PEG8 (28 atoms), PEG9 (31 atoms) and TTDS.

[0175] The peptides of the present invention comprise at least one C8-C 20 Generally, such fatty acids may be branched or cyclic. 20 The fatty acids are preferably attached to the N-terminus and / or C-terminus. 20 The fatty acid can be attached to any suitable functional group of the peptide and / or amino acid, such as a hydroxyl group, a carboxyl group, an amino group, or a thiol group, preferably an amino group or a carboxyl group. Preferably, the peptide of the present invention comprises one C8-C 20 Fatty acid, or one C8-C attached to the C-terminus 20 More preferably, one C8-C 20 The fatty acid is attached to the N- or C-terminus via an amide bond. 0 or X 15 The fatty acid side chain formed by the fatty acid is preferably a fatty acid > C8, and more preferably a fatty acid ≥ C 12 It is more preferable that the fatty acid is ≧C 14 It is more preferable that X 0 or X 15 The fatty acid side chain formed by 12-C 18 Fatty acids, preferably C 12 -C 16 Fatty acids, or C 14 -C 18 Fatty acids, or C 14 -C 16 More preferably, it is a fatty acid. Most preferably, it is a C 16 Fatty acids, such as palmitic acid (palmitoyl, palm) and C 18 Fatty acids, such as 1,18-octadecanedioic acid (ODD).

[0176] Furthermore, it is understood that the N- or C-terminal moieties may be linked, for example covalently linked, particularly in situations where the amino- or carboxy-terminus is attached to a linker or another chemical moiety.

[0177] Chemical groups, unnatural amino acids or moieties may be abbreviated herein as shown in Table 3.

[0178] Table 3: Abbreviations / expressions and nomenclature used for chemical groups, unnatural amino acids or further moieties in sequences [Table 3] TIFF0007667091000016.tif255166TIFF0007667091000017.tif254168TIFF000 7667091000018.tif254162TIFF0007667091000019.tif253168TIFF00076670910 00020.tif252166TIFF0007667091000021.tif252166TIFF0007667091000022.t if253168TIFF0007667091000023.tif250166TIFF0007667091000024.tif207167

[0179] In the definition of some portions of the peptides according to formula (I) or formula (II) of the present invention, the term "mimetic" used in connection with some amino acids refers to a respective amino acid mimic, such as an arginine mimic, an isoleucine mimic, or a proline mimic. Generally, a "protein mimic" refers to a molecule, such as a peptide, a modified peptide, or any other molecule, that biologically mimics the action or activity of some other protein. In connection with the use of the term "mimetic" in connection with a specific amino acid, the "mimetic" refers to any other amino acid, amino acid analog, amino acid derivative, amino acid conjugate, etc., that biologically mimics the action or activity of the respective amino acid.

[0180] Proline mimetics according to the invention are in particular (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, Hyp, morpholine-3-carboxylic acid, Pip, (4aR,6aR,9S,11aS)-11-oxo-2,3,4,4a,6a,7,8,9,11,11a-decahydro-1H-pyrido[3,2-e]pyrrolo[1,2-a]azepine-9-carboxylic acid or (trans-4-fluoro)P, (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, Oic, Hyp, (4-CF3)P, (cis-4-fluoro)P, 3,3-dimethyl-1,3-azasilolidine-5-carboxylic acid, (3S-OH)P, ( (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid or difluoroproline, (3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1), (3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2), and substituted proline.

[0181] Isoleucine mimetics according to the invention include, among others, (N-methyl)-I, allo-Ile, Cba, Nva, Abu, Leu, Cpg, cyclohexyl-Gly, (S)-2-amino-3-ethyl-pentanoic acid, 3-chloro-Phg, allo-Ile, Chg, cyclobutylglycine, allo-Ile, Cbg, (2S,3S)-2-((amino)methyl)-3-methylpentanoic acid), Phg, 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, 2-methyl-D-alloisoleucine, Nva, Abu, or Ala.

[0182] Leucine mimetics according to the invention include, in particular, (tBu)A, (2-chloro)F, (2-bromo)F, AAD, (2S)-2-amino-4,4,4-trifluorobutanoic acid, Cnba, (4-fluoro)L, (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propanoic acid, Gly(tBu), 3-(trimethylsilyl)-L-alanine, 2,5-difluoro-L-phenylalanine, 2-amino-7-(tert-butoxy)-7-oxoheptanoic acid, 5,5,5-trifluoro-L- Leucine ((trifluoro)L), (2-Me)F, Cba, Cpa, cyclopropylmethylalanine, trifluoromethylalanine or difluoromethylalanine, (2-fluoro)F, (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, (2S)-3-(3-cyanophenyl)-2-aminopropanoic acid, 2-amino-5,5,5-trifluoro-4-methyl-pentanoic acid, (2S)-2-amino-5-methyl-hexanoic acid or (2S)-3-(indol-4-yl)-2-(amino)propanoic acid.

[0183] The present invention further includes analogs and derivatives of the described peptides. The term "analog" or "derivative" of a peptide or amino acid sequence according to the present invention particularly includes any amino acid sequence that has an amino acid sequence identity of at least 80% or at least 85%, preferably at least 90%, more preferably at least 95%, and even more preferably at least 99% to the sequence and has the same or equivalent properties or activity. Sequence identity can be determined by common techniques (e.g., visual comparison) or by any computer tool commonly used in the art. An example is the BLAST program used with default parameters.

[0184] Analogs or derivatives of the peptides or amino acid sequences of the present invention may result from mutations or alterations in the sequence of the peptides of the present invention, including deletion or insertion of one or more amino acids, or substitution of one or more amino acids, or even alternative splicing. Several of these modifications may be combined. Preferably, analogs of the amino acid sequences of the present invention contain conservative substitutions in the amino acid sequence.

[0185] The term "conservative substitution" used herein refers to the substitution of one or more amino acids with another biologically similar residue. Examples include the substitution of amino acid residues with similar characteristics, such as small amino acids, acidic amino acids, polar amino acids, basic amino acids, hydrophobic amino acids and aromatic amino acids. For example, see the scheme in Table 4 below, in which conservative substitution of amino acids is classified according to physicochemical properties: I: neutral, hydrophilic; II: acid and amide; III: basic; IV: hydrophobic; V: aromatic, bulky amino acids; VI: neutral or hydrophobic; VII: acid; VIII: polar.

[0186] Table 4: Amino acids grouped according to their physicochemical properties [Table 4]

[0187] A peptide analog or derivative may also include one or more further modifications (eg, conjugation to another compound to form an amino acid conjugate).

[0188] Such modifications may alternatively or additionally involve modifying one or more amino acid residues in the peptides of the invention, e.g., chemical groups X 0 Or it may result from the conjugation of a chemical group, such as a PEG group, to the side chain as defined above in connection with the polymer moiety. Such modifications can, for example, increase the solubility and / or half-life and / or bioavailability of the peptide in vivo (e.g., in plasma), and are also known to reduce the clearance (e.g., renal clearance) of therapeutic proteins and peptides. Suitable modifications are well known to those skilled in the art and include, in particular, but not limited to, PEGylation of one or more side chains of the peptides of the present invention. Here, "PEGylation" refers to the act of coupling (e.g., covalently) a polyethylene glycol (PEG) structure to the peptide of the present invention. Those skilled in the art are familiar with possible PEGs for coupling to amino acid side chains of small peptides to form the respective conjugates, for example, from WO2015 / 200916A1 (which is incorporated herein by reference).

[0189] Unless otherwise specified, all peptides of the present invention are TFA salts. The present invention further includes pharmaceutically acceptable salts and salt-free forms of the peptides defined herein. Here, a pharmaceutically acceptable salt refers to a salt or zwitterionic form of the peptide or compound of the present invention that is water- or oil-soluble or dispersible, suitable for treating diseases without undue toxicity, irritation, or allergic reaction, commensurate with a reasonable benefit / risk ratio, and effective for its intended use. Salts can be prepared during the final isolation and purification of the compound, or separately by reacting the amino group with a suitable acid. Representative acid addition salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, carbonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate (isethionate), lactate, maleate, and methicone. Preferred acid addition salts include trifluoroacetate, formate, hydrochloride, and undecanoate.

[0190] In addition, the amino group in the compound of the present invention can be quaternized with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and steryl chlorides, bromides, and iodides; and benzyl bromide and phenethyl bromide. Examples of acids that can be used to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, and organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid. Suitable pharmaceutically acceptable salts can be, for example, salts selected from acid addition salts and base salts. Examples of acid addition salts include chloride salts, citrate salts, and acetate salts.

[0191] Examples of basic salts include those in which the cation is an alkali metal cation (e.g., sodium or potassium ion), an alkaline earth metal cation (e.g., calcium or magnesium ion), and a substituted ammonium ion (e.g., N(R 1 )(R 2 )(R 3 )(R 4 ) + Ions of the type, where R 1 , R 2 , R 3 and R 4 are each independently typically hydrogen, optionally substituted C 1-6 -Alkyl or optionally substituted C 2-6 -alkenyl). 1-6 Examples of -alkyl groups include methyl, ethyl, 1-propyl and 2-propyl groups. 2-6 Examples of -alkenyl groups include ethenyl, 1-propenyl and 2-propenyl, whereby salts in which the cation is selected from sodium, potassium and calcium are preferred.

[0192] Other examples of pharmaceutically acceptable salts are found in "Remington's Pharmaceutical Sciences," 17th ed., Alfonso R. Gennaro (ed.), Mark Publishing Company, Easton, PA, USA, 1985 (and more recent editions thereof), and "Encyclopaedia of Pharmaceutical Technology," 3rd ed., James Swarbrick (ed.), Informa Healthcare USA (Inc.), NY, USA, 2007. Also, for a review of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley VCH, 2002). Other suitable base salts are formed from bases that form non-toxic salts. Representative examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, and zinc salts, preferably choline. Hemisalts of acids and bases can also be formed, for example, hemisulfates and hemicalcium salts.

[0193] The present invention further includes solvates of the peptides defined herein. Here, the term "solvate" refers to a complex of defined stoichiometry formed between a solute (e.g., a peptide according to the present invention or its pharmaceutically acceptable salt) and a solvent. In this context, the solvent can be, for example, water, ethanol, or another pharmaceutically acceptable, typically small-molecule organic species (e.g., acetic acid or lactic acid, but not limited thereto). When the solvent in question is water, such a solvate is usually called a hydrate.

[0194] The compounds of the present invention have valuable pharmacological properties and can be used in the prevention and treatment of disorders in humans and animals.

[0195] In the context of the present invention, the term "treatment" or "treat" includes suppressing, slowing, arresting, ameliorating, attenuating, limiting, reducing, inhibiting, reversing or curing a disease, condition, disorder, injury or health condition, the development, course or progression of such condition and / or the symptoms of such condition (wherein the term "therapy" is understood to be synonymous with the term "treatment").

[0196] In the context of the present invention, the terms "prevention", "prophylaxis" or "precaution" are used interchangeably and refer to the avoidance or reduction of the risk of contracting, coming into contact with, suffering from or having a disease, condition, disorder, injury or health impairment, the onset or progression of such a condition, and / or the symptoms of such a condition.

[0197] Treatment or prevention of a disease, condition, disorder, injury or health impairment may be partial or complete.

[0198] The compounds of the present invention are particularly suitable for the treatment and / or prevention of cardiovascular, cardiopulmonary, renal, pulmonary, fibrotic, thromboembolic and inflammatory disorders.

[0199] The compounds according to the invention are therefore useful for treating, for example, cardiovascular and cardiopulmonary disorders and their sequelae, such as inflammatory heart disease, myocarditis, endocarditis, pericarditis, rheumatic fever with and without cardiac involvement, acute rheumatic pericarditis, acute rheumatic endocarditis, acute rheumatic myocarditis, chronic rheumatic heart disease with and without endocarditis, valvulitis, pericarditis, ischemic heart disease such as unstable angina and acute myocardial infarction, atrial and ventricular arrhythmias and conduction disorders, such as atrioventricular block of grades I to III, supraventricular tachyarrhythmias, atrial fibrillation, atrial flutter, ventricular fibrillation, ventricular flutter, ventricular tachyarrhythmias, torsades de pointes tachycardia, for the treatment and / or prevention of stroke due to intracerebral or intracranial hemorrhage, cerebral infarction following thromboembolic, arteriosclerotic, infectious, or inflammatory vascular lesions, tachycardia, atrial and ventricular premature contractions, AV-junction premature contractions, sick sinus syndrome, occlusion and stenosis of cerebral arteries (e.g., cerebral infarction following thromboembolic, arteriosclerotic, infectious, or inflammatory vascular lesions), peripheral ischemic tissue damage (e.g., arteriosclerotic gangrene) due to diseases of arteries, arterioles, and capillaries (e.g., thromboembolic, arteriosclerotic, infectious, or inflammatory vascular lesions, endarteritis deformans or endarteritis obliterans, aneurysmal dissections) for the treatment and / or prevention of postoperative disorders of the circulatory system, such as systemic inflammatory response syndrome, postoperative vasoplegia, post-cardiotomy syndrome, postoperative hypotension and heart failure, for the prevention and treatment of ischemia-reperfusion injury and organ dysfunction after thrombolytic therapy, percutaneous transluminal angioplasty (PTA), percutaneous transluminal coronary angioplasty (PTCA), bypass surgery and heart, lung, liver and kidney transplantation, and for the treatment and / or prevention of delayed graft function after kidney transplantation.

[0200] The compounds according to the invention are further suitable for the treatment of shock, such as cardiogenic shock, septic shock and anaphylactic shock, by preventing MASP-mediated end-organ damage.

[0201] Furthermore, the compounds of the present invention have anti-inflammatory properties and can therefore be used as anti-inflammatory agents for the treatment and / or prevention of sepsis (SIRS), multiple organ failure (MODS, MOF), inflammatory disorders of the kidney, chronic intestinal inflammation (IBD, Crohn's disease, UC), pancreatitis, peritonitis, rheumatic disorders, inflammatory skin disorders and inflammatory eye disorders.

[0202] Due to their activity profile, the compounds of the invention are particularly suitable for the treatment and / or prevention of the cardiovascular, pulmonary, cerebral and renal sequelae of sepsis and systemic inflammatory response syndrome.

[0203] The compounds according to the invention are particularly suitable for the treatment and / or prevention of ischemia and / or reperfusion-related damage to the heart and kidneys and other organs in the context of resuscitation and surgical interventions such as, but not limited to, bypass surgery, heart valve surgery, and aortic aneurysm surgery.

[0204] The compounds according to the invention can furthermore be used to prevent ischemia and / or reperfusion-related damage to organs or tissues, and also as additives for perfusion and preservation solutions of organs, organ parts, tissues or tissue parts of human or animal origin, in particular for surgical interventions or in the field of transplant medicine.

[0205] Furthermore, the compounds according to the invention are suitable for the treatment and / or prevention of diseases of the blood and blood-forming organs and immune system, including, but not limited to, acquired hemolytic anemia, hemolytic uremic syndrome, paroxysmal nocturnal hemoglobinuria (Marchiafava-Micheli), coagulation deficiencies, purpura and other bleeding conditions, disseminated intravascular coagulation (defibrination syndrome), essential (hemorrhagic) thrombocythemia, purpura fulminans, thrombotic thrombocytopenic purpura, allergic purpura, allergic vasculitis, lymphopenia and granulocytosis, and sarcoidosis.

[0206] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of diabetic sequelae, such as diabetic renal complications, diabetic nephropathy, intracapillary glomerular nephropathy, diabetic ocular complications, diabetic retinopathy, neurological complications, diabetic polyneuropathy, and cardiovascular complications such as microangiopathy and gangrene.

[0207] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of inflammatory diseases of the nervous system such as multiple sclerosis, meningitis and encephalitis, bacterial and viral meningitis and encephalitis, post-immune encephalitis, inflammatory polyneuropathies, and polyneuropathies in infectious and parasitic diseases.

[0208] The compounds according to the invention are further useful in the treatment of diseases of the eye and its adnexa, such as acute and subacute iridocyclitis, choroidal degeneration, chorioretinal inflammation, chorioretinal inflammation in infectious and parasitic diseases, background retinopathies and retinal vascular changes, proliferative retinopathies, macular degeneration and posterior pole, peripheral retinal degeneration, age-related macular degeneration (AMD), including dry (non-exudative) and wet (exudative, neovascular) AMD, choroidal neovascularization (CNV), choroidal neovascular membrane (CNVM), It is suitable for the treatment and / or prevention of cystoid macular edema (CME), epiretinal membrane (ERM) and macular perforation, choroidal neovascularization associated with myopia, pigmented streaks and retinal detachment, diabetic retinopathy, diabetic macular edema (DME), atrophic and hypertrophic lesions of the retinal pigment epithelium, retinal vein occlusion, choroidal retinal vein occlusion, macular edema, macular edema associated with retinal vein occlusion, post-operative disorders of the eye and adnexa, e.g. keratopathy after cataract surgery.

[0209] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of diseases of the respiratory system, including but not limited to viral, bacterial and fungal pneumonias, radiation pneumonitis, pneumoconiosis, allergic alveolitis, airway diseases due to certain organic dusts, e.g. farmer's lung due to chemicals, gases, fumes and vapors, bronchitis, pneumonitis and pulmonary edema, drug-induced interstitial lung disorders, adult respiratory distress syndrome (ARDS) and acute lung injury (ALI), acute edema of the lung, interstitial lung diseases with fibrosis, rheumatic lung diseases, respiratory disorders in other diffuse connective tissue disorders, e.g. systemic lupus erythematosus, scleroderma and Wegener's granulomatosis.

[0210] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of microvascular damage, thrombosis and subsequent thromboembolic events caused by viral infections such as, but not limited to, influenza viruses (e.g., caused by strains of serotypes H1N1, H5N1, H7N9), and coronaviruses (e.g., SARS-CoV, the etiological agent of Severe Acute Respiratory Syndrome (SARS), MERS-CoV, the etiological agent of Middle East Respiratory Syndrome (MERS), and SARS-CoV-2, the etiological agent of the COVID-19 pandemic).

[0211] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of diseases of the digestive system, including, but not limited to, non-infectious enteritis and colitis, such as Crohn's disease and ulcerative colitis, pancreatitis (including acute alcoholic and drug-induced pancreatitis), cholecystitis, inflammatory liver diseases, hepatorenal syndrome, post-operative damage to the liver, e.g. after liver surgery.

[0212] Due to their activity profile, the compounds of the present invention are particularly suitable for the treatment and / or prevention of diseases of the genitourinary system, including, but not limited to, acute renal failure, acute kidney injury (AKI), surgery-related AKI, sepsis-related AKI, contrast- and chemotherapy-induced AKI, renal ischemia and infarction, complications such as hypersensitivity in connection with hemodialysis and hemodiafiltration, cystitis, radiation cystitis, inflammatory diseases of the prostate, and endometriosis.

[0213] The compounds according to the invention are further suitable for the treatment and / or prevention of the sequelae of burns and injuries, including but not limited to early complications of trauma, traumatic anuria, crush syndrome, renal failure after crush, traumatic ischemia of muscles, traumatic brain injury, organ damage after exposure to electric current, radiation and extreme ambient air temperatures and pressures, after exposure to smoke, fire and flames, after contact with toxic animals and plants.

[0214] Due to their activity profile, the compounds of the invention are further suitable for the treatment of inflammatory skin diseases, e.g. epidermolytic skin diseases such as cutaneous lupus erythematosus, bullous disorders and pemphigus subtypes, papulosquamous disorders such as psoriasis, dermatitis and eczema, urticaria and erythema.

[0215] According to a further embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt or solvate thereof comprising, consisting essentially of or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, for use in the prevention and / or treatment of disease.

[0216] According to a further embodiment, the present invention provides a compound comprising a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt or solvate thereof, which comprises, consists essentially of or consists of formula (I) or formula (II) and may be isolated and / or purified, for use in the prevention and / or treatment of MASP-associated disorders.

[0217] According to a further embodiment, the present invention provides a peptide-containing compound or derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or solvate of a salt thereof comprising, consisting essentially of or consisting of formula (I) or formula (II), which may be isolated and / or purified, for use in the prevention and / or treatment of MASP-associated disorders, which acts as a MASP-1 and / or MASP-2 inhibitor and / or inhibits C3 deposition.

[0218] According to further embodiments, the present invention provides compounds containing a peptide or derivatives, prodrugs, analogs, pharmaceutically acceptable salts or solvates thereof comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, for use in the prevention and / or treatment of cardiovascular and cardiopulmonary disorders, crush shock, inflammatory disorders, sequelae of cardiovascular, pulmonary, cerebral and renal sepsis, ischemia and / or reperfusion-related injury, acute kidney injury, graft protection and delayed graft function, diseases of the blood and blood-forming organs and immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, diseases of the respiratory, digestive or genitourinary systems, and sequelae of burns and injuries.

[0219] According to further embodiments, the present invention provides compounds containing peptides or derivatives, prodrugs, analogs, pharmaceutically acceptable salts or solvates thereof comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, for use in the prevention and / or treatment of diseases of the genitourinary system, including, but not limited to, acute renal failure, acute kidney injury (AKI), surgery-related AKI, sepsis-related AKI, contrast and chemotherapy-induced AKI, renal ischemia and infarction, complications such as hypersensitivity associated with hemodialysis and hemodiafiltration, cystitis, radiation cystitis, inflammatory disease of the prostate, and endometriosis.

[0220] The present invention further relates to a method for treating or ameliorating a MASP-associated disorder as defined above in a subject or patient in need thereof by administering to the subject or patient in need thereof at least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, a complex or pharmaceutical composition as defined above.

[0221] As used herein, the terms "patient," "subject," or "individual" are used interchangeably and can refer to either a human or a non-human animal. These terms include mammals such as humans, primates, livestock animals (e.g., cows, pigs), companion animals (e.g., dogs, cats), and rodents (e.g., mice and rats). The term "mammal" refers to any mammalian species, such as humans, mice, rats, dogs, cats, hamsters, guinea pigs, rabbits, livestock, etc.

[0222] According to the present invention, at least one peptide, derivative, or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a conjugate as defined above, is administered to a patient or subject in a therapeutically effective amount, wherein a "therapeutically effective amount" of a compound of the present invention is meant to describe a sufficient amount of a compound of the present invention to treat a MASP-associated disorder as defined herein. In certain embodiments, the therapeutically effective amount achieves a desired benefit / risk ratio applicable to any medical treatment.

[0223] The present invention particularly includes the following embodiments, in which the substituents or moieties of the peptides defined in formula (I) or formula (II) may independently have the meanings as described below.

[0224] A peptide or a compound comprising a peptide, or a derivative or analogue thereof as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex or pharmaceutical composition (defined below), is hereinafter also generally referred to as a "MASP inhibitory peptide of the present invention."

[0225] In some embodiments, the MASP inhibitory peptides of the present invention bind to MASP-1 and / or MASP-2, e.g., human MASP-1 and / or MASP-2. In certain embodiments, the MASP inhibitory peptides of the present invention specifically bind to human MASP-1 and / or MASP-2. As used herein, "specifically bind" refers to the preferential interaction of a specific binding agent with a given ligand over other agents in a sample. For example, a specific binding agent that specifically binds to a given ligand binds to the given ligand in an amount or degree that is observable above the amount or degree of any non-specific interactions with other components in the sample under appropriate conditions. Suitable conditions are those that allow for the interaction between a given specific binding agent and a given ligand. These conditions include pH, temperature, concentration, solvent, incubation time, etc., and may vary between a given specific binding agent and ligand pair but can be easily determined by one of skill in the art. In some embodiments, the MASP inhibitory peptides of the present invention bind to MASP-1 and / or MASP-2 with greater specificity than a MASP inhibitory peptide reference compound (e.g., any one of the MASP inhibitory peptide reference compounds provided herein).

[0226] Thus, the present invention further relates to a complex comprising at least one peptide or compound containing a peptide, derivative or analogue as defined herein, bound to MASP-1 or MASP-2.

[0227] In some embodiments, the MASP inhibitory peptides of the present invention exhibit specific binding to MASP-1 and / or MASP-2, particularly human MASP-1 and / or MASP-2, i.e., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% higher than the selected MASP inhibitory peptide reference compound.

[0228] In some embodiments, the MASP inhibitory peptides of the present invention exhibit specific binding to MASP-1 and / or MASP-2, particularly human MASP-1 and / or MASP-2, that is at least about 1, 2, 3, 4, 5 times, or at least about 10, 20, 50, or 100 times higher than the selected MASP inhibitory peptide reference compound.

[0229] In some embodiments, the MASP inhibitory peptides of the present invention exhibit binding affinity for MASP-1 and / or MASP-2 that is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% higher than the selected MASP inhibitory peptide reference compound.

[0230] In some embodiments, the MASP inhibitory peptides of the present invention exhibit binding affinity for MASP-1 and / or MASP-2 that is at least about 1, 2, 3, 4, 5 times, or at least about 10, 20, 50, 100, or 1000 times higher than the selected MASP inhibitory peptide reference compound.

[0231] In some embodiments, the MASP inhibitory peptides of the present invention exhibit inhibition of MASP-1 and / or MASP-2 (e.g., rat or human MASP-1 and / or MASP-2) activity. In some embodiments, the activity is an in vitro or in vivo activity, such as an in vitro or in vivo activity described herein. In some embodiments, the MASP inhibitory peptides of the present invention inhibit at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% of the MASP-1 and / or MASP-2 activity inhibited by the selected MASP inhibitory peptide reference compound.

[0232] In certain embodiments, the MASP inhibitory peptides of the present invention exhibit 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, or 200 times greater inhibition of MASP-1 and / or MASP-2 than the selected MASP inhibitory peptide reference compound.

[0233] In a further specific embodiment, the MASP-1 and / or MASP-2 inhibitory activity of the MASP inhibitory peptides of the present invention is measured by the IC 50 It is determined by measuring the IC against MASP-1 and / or MASP-2. 50 Determination of IC for MASP-1 and / or MASP-2 can be performed using the biochemical assays set forth herein. The MASP inhibitory peptides of the present invention have an IC for MASP-1 and / or MASP-2 of <1,000 nM, preferably ≦500 nM, more preferably ≦300 nM, more preferably ≦250 nM, more preferably ≦200 nM, more preferably ≦150 nM, more preferably ≦100 nM, more preferably ≦75 nM, more preferably ≦50 nM, more preferably ≦45 nM, more preferably ≦40 nM, more preferably ≦35 nM, more preferably ≦30 nM. 50 It is particularly preferred that

[0234] In some embodiments, the MASP inhibitory peptides of the present invention have a lower IC against MASP-1 and / or MASP-2 (e.g., rat or human MASP-1 and / or MASP-2) compared to a selected MASP inhibitory peptide reference compound. 50In some embodiments, the MASP inhibitory peptides of the present invention have an IC that is at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000% or 10000% lower than that of the selected MASP inhibitory peptide reference compound in a MASP-1 and / or MASP-2 competitive binding assay. 50 It has.

[0235] In some embodiments, the MASP inhibitory peptides of the present invention exhibit in vitro inhibition of human MASP-1 and / or MASP-2 activity of at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or greater than 99%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% greater than that of the selected MASP inhibitory peptide reference compound.

[0236] In some embodiments, the MASP inhibitory peptides of the present invention exhibit in vivo inhibition of human MASP-1 and / or MASP-2 activity of greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or 99%, 100%, 200%, 300%, 400%, 500%, 700%, 1000% or 10,000% greater than that of the selected MASP inhibitory peptide reference compound.

[0237] As used herein, in certain embodiments, a MASP inhibitory peptide having "MASP-1 and / or MASP-2 inhibitory activity" means that the compound has the ability to inhibit C3 deposition in a dose- and time-dependent manner in vitro or in a subject (e.g., a mouse or human) when administered thereto (e.g., by a parenteral route, e.g., by injection, or by the pulmonary, nasal, sublingual, lingual, oral, cutaneous, transdermal, conjunctival, or ocular route, or as an implant or stent administered orally).

[0238] In some embodiments, the MASP inhibitory peptides of the present invention exhibit inhibition of C3 deposition (e.g., human C3 deposition). In some embodiments, the inhibition of C3 deposition is determined by in vitro or in vivo inhibition. In some embodiments, the MASP inhibitory peptides of the present invention inhibit at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% of the C3 deposition inhibited by the selected MASP inhibitory peptide reference compound.

[0239] In certain embodiments, the MASP inhibitory peptides of the present invention exhibit inhibition of C3 deposition that is 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, or 200 times greater than the selected MASP inhibitory peptide reference compound.

[0240] In a further particular embodiment, the MASP-1 and / or MASP-2 inhibitory activity of the MASP inhibitory peptides according to the invention is measured by their IC for inhibition of C3 deposition in vitro or in a subject (e.g., mouse or human). 50 The IC for C3 deposition is determined by measuring 50 Determination of IC for C3 deposition can be performed using the C3 human deposition assay provided herein. The MASP inhibitory peptides of the present invention have an IC for C3 deposition of <1,000 nM, preferably ≦500 nM, more preferably ≦300 nM, more preferably ≦250 nM, more preferably ≦200 nM, more preferably ≦150 nM, more preferably ≦100 nM, more preferably ≦75 nM, more preferably ≦50 nM, more preferably ≦45 nM, more preferably ≦40 nM, more preferably ≦35 nM, more preferably ≦30 nM. 50 It is particularly preferred that

[0241] In some embodiments, the MASP inhibitory peptides of the present invention exhibit in vitro inhibition of C3 deposition of at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or greater than 99%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% greater than that of the selected MASP inhibitory peptide reference compound.

[0242] In some embodiments, the MASP inhibitory peptides of the present invention exhibit in vivo inhibition of C3 deposition of at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or greater than 99%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10,000% greater than that of the selected MASP inhibitory peptide reference compound.

[0243] It is particularly preferred that the peptides according to the invention act as MASP inhibitory peptides, the activity of which is determined according to at least one of the specific assays and / or in vivo studies according to the embodiments of the present invention.

[0244] Due to their aforementioned MASP-1 and / or MASP-2 inhibitory activity and C3 deposition inhibitory activity, compounds containing the peptides of the present invention or the peptides of the present invention (including their analogs, derivatives, and pharmaceutically acceptable salts or solvates thereof, as well as the aforementioned complexes) are suitable for use in the prevention and / or treatment of MASP-1 and / or MASP-2 associated disorders.

[0245] According to a further embodiment, the present invention provides compounds comprising a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or solvate of a salt thereof, which comprises, consists essentially of or consists of formula (I) or formula (II) and which may be isolated and / or purified, which act as MASP-1 and / or MASP-2 inhibitors and / or inhibit C3 deposition.

[0246] The compound of the present invention can be used alone or in combination with other active compounds as needed.The present invention also relates to a medicament that contains at least one compound according to the present invention and one or more additional active compounds, particularly for the treatment and / or prevention of the above-mentioned diseases.As suitable combined active compounds, for example and preferably, the following can be mentioned: compounds that inhibit the breakdown of cyclic guanosine monophosphate (cGMP) and / or cyclic adenosine monophosphate (cAMP), such as inhibitors of phosphodiesterase (PDE) 1, 2, 3, 4 and / or 5, in particular PDE4 inhibitors such as roflumilast or levamilast and PDE5 inhibitors such as sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, mirodenafil or lodenafil; NO-independent but heme-dependent stimulators of guanylate cyclase, in particular riociguat, nerociguat, veruiciguat and the compounds described in WO00 / 06568, WO00 / 06569, WO02 / 42301, WO03 / 095451, WO2011 / 147809, WO2012 / 004258, WO2012 / 028647 and WO2012 / 059549; NO-independent and heme-independent activators of guanylate cyclase, in particular lancaciguat, BI703704 and compounds described in WO2012 / 139888, WO2001 / 019780 and WO2014 / 012934; prostacyclin analogues and IP receptor agonists, for example and preferably iloprost, beraprost, treprostinil, epoprostenol, NS-304, selexipag, or ralinepag; Endothelin receptor antagonists, for example and preferably bosentan, darusentan, ambrisentan, macisentan or sitaxsentan; · Vasopressin receptor antagonists, such as tolvaptan, conivaptan, and lercovaptan; Human neutrophil elastase (HNE) inhibitors, for example and preferably sivelestat or DX-890 (Reltran); compounds that inhibit signal transduction cascades, in particular from the group of the tyrosine kinase inhibitors, such as, for example and preferably, dasatinib, nilotinib, bosutinib, regorafenib, sorafenib, sunitinib, cediranib, axitinib, telatinib, imatinib, brivanib, pazopanib, vatalanib, gefitinib, erlotinib, lapatinib, canertinib, lestaurtinib, pelitinib, semaxanib, masitinib or tandutinib; · Signal transduction modulators from the ASK1 kinase inhibitor group, such as selonsertib; Rho kinase inhibitors, for example and preferably fasudil, Y-27632, SLx-2119, BF-66851, BF-66852, BF-66853, KI-23095 or BA-1049; · Active ingredients that reduce the permeability of the vascular wall (edema formation), for example and by way of preferential inhibitors of the ALK1-Smad1 / 5 signaling pathway, inhibitors of the VEGF and / or PDGF signaling pathway, cyclooxygenase inhibitors, inhibitors of the kallikrein-kinin system or inhibitors of the sphingosine-1-phosphate signaling pathway; · Corticosteroids, such as cortisone, cortisol, prednisolone, methylprednisolone, triamcinolone, or dexamethasone; · Active ingredients that reduce damage to organs under oxidative stress, such as, for example, inhibitors of the complement system, in particular antagonists of the complement C5a receptor, anti-C5 antibodies or agonists of the 5-HT1A receptor; Modulators, stimulators and enhancers of the transcription factor Nrf2, such as CXA-10, oltipraz, dimethyl fumarate or bardoxolone; Adrenomedullin and adrenomedullin derivatives, such as pegylated adrenomedullin, and adrenomedullin stabilizers, such as adrecizumab; · Compounds that inhibit hypoxia-inducible factor prolyl hydroxylases (HIF-PH inhibitors), such as molidusat, vadadustat, roxadustat, daprodustat, or desidastat; · Compounds that inhibit the induction of cell death and apoptotic pathways, such as QPI-1002; C-Met agonists and hepatocyte growth factor mimetics, such as refanalin; · Alkaline phosphatase and recombinant alkaline phosphatase; · Compounds that inhibit inflammatory responses and T-cell proliferation, for example, CD28 antagonistic compounds such as reltesimod; · Compounds that modulate the activation of Th17 T cells, such as modulators of the RORc / ROR-gamma transcription factor; · Compounds that counteract Th17 T cell responses, for example anti-IL-17 and anti-IL-23 antibodies, such as ixekizumab, secukinumab, brodalumab, ustekinumab, geselkumaab or PTG-200; antithrombotic agents, for example preferably from the group of platelet aggregation inhibitors, anticoagulants or profibrinolytic substances; In a preferred embodiment of the invention, the compounds of the invention are administered in combination with a platelet aggregation inhibitor, such as, for example and preferably, aspirin, clopidogrel, ticlopidine or dipyridamole.

[0247] In a preferred embodiment of the invention, the compounds of the invention are administered in combination with a thrombin inhibitor, such as, for example and preferably, ximelagatran, melagatran, dabigatran, bivalirudin or clexane.

[0248] In a preferred embodiment of the invention, the compounds of the invention are administered in combination with a GPIIb / IIIa antagonist, such as, for example and preferably, tirofiban or abciximab.

[0249] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a factor Xa inhibitor, for example and preferably rivaroxaban, apixaban, fidexaban, razaxaban, fondaparinux, idraparinux, DU-176b, PMD-3112, YM-150, KFA-1982, EMD-503982, MCM-17, MLN-1021, DX9065a, DPC906, JTV803, SSR-126512 or SSR-128428.

[0250] In a preferred embodiment of the invention, the compounds of the invention are administered in combination with heparin or a low molecular weight (LMW) heparin derivative.

[0251] In a preferred embodiment of the invention, the compounds of the invention are administered in combination with direct inhibitors of coagulation factor XI, inhibitors of coagulation factor XI expression, and anti-coagulation factor XI antibodies (e.g., xysomab 3G3); In a preferred embodiment of the invention, the compounds of the invention are administered in combination with a mineralocorticoid receptor antagonist, such as, for example and preferably, spironolactone, eplerenone or finerenone.

[0252] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a diuretic, for example and preferably furosemide, bumetanide, torsemide, bendroflumethiazide, chlorthiazide, hydrochlorthiazide, hydroflumethiazide, methylclothiazide, polythiazide, trichlormethiazide, chlorthalidone, indapamide, metolazone, quinethazone, acetazolamide, dichlorphenamide, methazolamide, glycerol, isosorbide, mannitol, amiloride or triamterene.

[0253] In a preferred embodiment of the invention, the compounds of the invention are administered in combination with a PPAR-gamma agonist, such as, for example and preferably, pioglitazone or rosiglitazone.

[0254] In a preferred embodiment of the invention, the compounds of the invention are administered in combination with a PPAR-delta agonist, for example and preferably GW501516 or BAY68-5042.

[0255] According to a further embodiment, the present invention provides pharmaceutical compositions comprising at least one compound containing a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt, solvate or solvate of a salt thereof comprising, consisting essentially of or consisting of formula (I) or formula (II), which may be isolated and / or purified, in combination with one or more further active ingredients selected from the group consisting of inhibitors of phosphodiesterase, stimulators or activators of guanylate cyclase, IP receptor agonists, mineralocorticoid receptor antagonists, diuretics, PPAR-gamma agonists, PPAR-delta agonists, corticosteroids, active ingredients which reduce organ damage under oxidative stress, compounds which inhibit the induction of cell death and apoptosis pathways, compounds which inhibit inflammatory responses and T-cell proliferation, antithrombotic, platelet aggregation inhibitors, thrombin inhibitors, GPIIb / IIIa antagonists, factor Xa inhibitors, heparin or low molecular weight (LMW) heparin derivatives and inhibitors of coagulation factor XI.

[0256] The present invention further relates to a kit-of-parts combination comprising at least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, a conjugate or pharmaceutical composition as defined above, and at least one selected from a reagent, a medical device, an instruction manual, or any combination thereof.

[0257] The present invention further relates to a medical device comprising at least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, a complex or a pharmaceutical composition as defined above, for delivery of the peptide, derivative, analogue or complex thereof, or a pharmaceutical composition thereof to a subject.

[0258] The pharmaceutical composition, kit-of-parts combination or medical device as defined above is particularly intended for use in the prevention and / or treatment of a disorder or disease as defined herein.

[0259] The MASP inhibitory peptides of the present invention can have systemic and / or local activity, and to this end, they can be administered in any suitable manner, for example, via oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, vaginal, cutaneous, transdermal, conjunctival, otic routes, or as an implant or stent.

[0260] For these administration routes, the compounds according to the invention can be administered in suitable dosage forms.

[0261] For oral administration, the compounds of the present invention can be formulated into dosage forms known in the art that deliver the compounds of the present invention rapidly and / or in a modified manner, such as tablets (uncoated or coated tablets, e.g., tablets with enteric or controlled-release coatings that dissolve slowly or are insoluble), orally disintegrating tablets, films / wafers, films / lyophilisates, capsules (e.g., hard or soft gelatin capsules), dragees, granules, pellets, powders, emulsions, suspensions, aerosols, or solutions. The compounds of the present invention can be incorporated into such dosage forms in crystalline and / or amorphous and / or dissolved form.

[0262] Parenteral administration can be carried out by bypassing the absorption step (for example, intravenously, intraarterially, intracardially, intraspinally or intralumbarly) or by encapsulating the absorption step (for example, intramuscularly, subcutaneously, intradermally, transdermally or intraperitoneally, intraocularly). Suitable dosage forms for parenteral administration are, inter alia, preparations for injection and infusion in the form of solutions, suspensions, emulsions, lyophilisates or sterile powders.

[0263] Examples of suitable forms for other administration routes are pharmaceutical forms for inhalation (especially powder inhalers, nebulizers), nasal drops, nasal solutions, nasal sprays; tablets / films / wafers / capsules for lingual, sublingual or buccal administration; suppositories; eye drops, eye ointments, eye baths, eye inserts, eye drops, ear drops, ear sprays, ear powders, ear rinses, ear tampons; vaginal capsules, topical applications, aqueous suspensions (lotions, mixturae agitandae), lipophilic suspensions, emulsions, ointments, creams, transdermal therapeutic systems (e.g. patches), milks, pastes, foams, dusting powders, implants or stents.

[0264] According to a further embodiment, the present invention provides pharmaceutical compositions comprising at least one compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt thereof, comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, in combination with one or more inert, non-toxic, pharmaceutically suitable excipients.

[0265] The compounds according to the invention can be incorporated into the dosage forms described. This can be achieved by mixing them with pharmaceutically suitable excipients in a manner known per se. Pharmaceutically suitable excipients include, inter alia: Fillers and carriers (e.g., cellulose, microcrystalline cellulose (e.g., Avicel®), lactose, mannitol, starch, calcium phosphate (e.g., Di-Cafos®)), Ointment bases (e.g., petroleum jelly, paraffin, triglycerides, waxes, wool wax, wool wax alcohol, lanolin, hydrophilic ointments, polyethylene glycol), suppository bases (e.g., polyethylene glycol, cocoa butter, hard fats), solvents (e.g., water, ethanol, isopropanol, glycerol, propylene glycol, medium-chain triglyceride fatty oils, liquid polyethylene glycol, paraffin); surfactants, emulsifiers, dispersing agents or wetting agents (e.g. sodium dodecyl sulfate), lecithins, phospholipids, fatty alcohols (e.g. Lanette®), sorbitan fatty acid esters (e.g. Span®), polyoxyethylene sorbitan fatty acid esters (e.g. Tween®), polyoxyethylene fatty acid glycerides (e.g. Cremophor®), polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, glycerol fatty acid esters, poloxamers (e.g. Pluronic®), buffers, acids and bases (e.g., phosphates, carbonates, citric acid, acetic acid, hydrochloric acid, sodium hydroxide solution, ammonium carbonate, trometamol, triethanolamine); Isotonic agents (glucose, sodium chloride, etc.), Adsorbents (e.g., highly dispersed silica), thickening agents, gel-forming agents, thickeners and / or binders (e.g., polyvinylpyrrolidone, methylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, starch, carbomer, polyacrylic acid (e.g., Carbopol®); alginates, gelatin), disintegrants (e.g., modified starch, sodium carboxymethylcellulose, sodium starch glycolate (e.g., Explotab®), cross-linked polyvinylpyrrolidone, croscarmellose sodium (e.g., AcDiSol®)); flow regulators, lubricants, glidants and release agents (e.g. magnesium stearate, stearic acid, talc, highly dispersed silica (e.g. Aerosil®)); coating materials (e.g. sugar, shellac) and film-forming agents for rapidly or modified dissolving films or diffusion membranes (e.g. polyvinylpyrrolidone (e.g. Kollidon®), polyvinyl alcohol, hydroxypropyl methylcellulose, hydroxypropyl cellulose, ethyl cellulose, hydroxypropyl methylcellulose phthalate, cellulose acetate, cellulose acetate phthalate, polyacrylates, polymethacrylates, e.g. Eudragit®), capsule materials (e.g., gelatin, hydroxypropyl methylcellulose), synthetic polymers (e.g. polylactides, polyglycolides, polyacrylates, polymethacrylates (e.g. Eudragit®), polyvinylpyrrolidones (e.g. Kollidon®), polyvinyl alcohols, polyvinyl acetates, polyethylene oxides, polyethylene glycols and their copolymers and block copolymers); Plasticizers (e.g., polyethylene glycol, propylene glycol, glycerol, triacetin, triacetyl citrate, dibutyl phthalate), penetration enhancers, stabilizers (e.g., antioxidants, e.g., ascorbic acid, ascorbyl palmitate, sodium ascorbate, butylhydroxyanisole, butylhydroxytoluene, propyl gallate), Preservatives (e.g., parabens, sorbic acid, thiomersal, benzalkonium chloride, chlorhexidine acetate, sodium benzoate), colorants (e.g. inorganic pigments, e.g. iron oxide, titanium dioxide), Flavorings, sweeteners, flavorings and / or odor masking agents.

[0266] Furthermore, the present invention relates to pharmaceutical compositions comprising at least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex as defined above.

[0267] In particular, the present invention relates to pharmaceutical compositions comprising at least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex as defined above, conventionally together with one or more pharmaceutically suitable excipients, and their use according to the invention.

[0268] The pharmaceutical composition according to the invention may comprise at least one additional active ingredient, such as an additional active ingredient which is preferably active in the prevention and / or treatment of a disorder or disease as defined herein.

[0269] At least one peptide, derivative or analogue as defined herein, or its pharmaceutically acceptable salt or solvate, or the conjugate or pharmaceutical composition as defined above, can be administered enterally or parenterally (including intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, intradermal and intraarticular injection and infusion), orally, vaginally, intraperitoneally, rectally, topically or buccally. Suitable formulations for each administration route are well known to those skilled in the art and include, but are not limited to, pills, tablets, enteric-coated tablets, film tablets, layer tablets, sustained-release or sustained-release preparations for oral administration, plasters, topical sustained-release preparations, dragees, pessaries, gels, ointments, syrups, granules, suppositories, emulsions, dispersions, microcapsules, microformulations, nanoformulations, liposomal preparations, capsules, enteric-coated capsules, powders, inhalation powders, microcrystalline preparations, inhalation sprays, powders, drops, nasal drops, nasal sprays, aerosols, ampoules, solutions, juices, suspensions, infusion or injection solutions, etc.

[0270] According to further embodiments, the present invention provides pharmaceutical compositions comprising at least one compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt thereof comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, in combination with one or more inert, non-toxic, pharmaceutically suitable excipients for use in the prevention and / or treatment of cardiovascular and cardiopulmonary disorders, crush injury, inflammatory disorders, sequelae of cardiovascular, pulmonary, cerebral and renal sepsis, ischemia and / or reperfusion-related injury, acute kidney injury, graft protection and delayed graft function, diseases of the blood and blood-forming organs and immune system, sequelae of diabetes, inflammatory diseases of the nervous system, diseases of the eye, diseases of the skin, diseases of the respiratory, digestive or genitourinary systems, sequelae of burns and trauma.

[0271] According to further embodiments, the present invention provides an anti-inflammatory drug comprising an inhibitor of phosphodiesterase, a stimulator or activator of guanylate cyclase, an IP receptor agonist, a mineralocorticoid receptor antagonist, a diuretic, a PPAR-gamma agonist, a PPAR-delta agonist, a corticosteroid, an oxidant, ... oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an oxidant, an Provided are pharmaceutical compositions comprising at least one compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt thereof comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which can be isolated and / or purified, in combination with one or more further active ingredients selected from the group consisting of active ingredients that reduce damage to organs under stress, compounds that inhibit the induction of cell death and apoptotic pathways, compounds that inhibit inflammatory responses and T-cell proliferation, antithrombotic agents, platelet aggregation inhibitors, thrombin inhibitors, GPIIb / IIIa antagonists, factor Xa inhibitors, heparin or low molecular weight (LMW) heparin derivatives and inhibitors of coagulation factor XI.

[0272] According to a further embodiment, the present invention provides a method for the treatment of a rheumatoid arthritis by administering an effective amount of a pharmaceutical composition comprising at least one compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt thereof, which comprises, consists essentially of, or consists of Formula (I) or Formula (II), which may be isolated and / or purified, or a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt thereof, which comprises, consists essentially of, or consists of Formula (I) or Formula (II), which may be isolated and / or purified. The present invention provides methods for treating and / or preventing cardiovascular and cardiopulmonary disorders, crush injury, inflammatory disorders, sequelae of cardiovascular, pulmonary, cerebral and renal sepsis, ischemia and / or reperfusion-related injury, acute kidney injury, graft protection and delayed graft function, diseases of the blood and blood-forming organs and immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, diseases of the respiratory system, digestive system or genitourinary system, burns and sequelae of trauma in humans and animals by administering an effective amount of a pharmaceutical composition containing at least one compound comprising the compound of the present invention in combination with one or more inert, non-toxic, pharmaceutically suitable excipients.

[0273] The appropriate dose of the MASP inhibitor peptide of the present invention can be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend on various factors, including: a) the disorder being treated and the severity of the disorder; b) the activity of the specific compound used; c) the specific composition used, the patient's age, weight, general health, sex, and diet; d) the time of administration, route of administration, and excretion rate of the specific hepcidin analog used; e) the duration of treatment; f) drugs used in combination with or simultaneously with the MASP inhibitor peptide used, and similar factors well known in the medical field.

[0274] In certain embodiments, the total daily dose of the MASP inhibitory peptide of the present invention administered to a subject or patient in a single or divided dose is, for example, 0.0001 to 300 mg / kg body weight / day or 1 to 300 mg / kg body weight / day, or about 0.0001 to about 100 mg / kg body weight / day, for example, about 0.0005 to about 50 mg / kg body weight / day, for example, about 0.001 to about 10 mg / kg body weight / day, for example, about 0.01 to about 1 mg / kg body weight / day, and can be administered in one or more doses, for example, 1 to 3 doses. Generally, the MASP inhibitory peptide of the present invention can be administered continuously (e.g., by intravenous administration or another continuous drug administration method) or can be administered to a subject at intervals, typically at regular time intervals, depending on the desired dosage and pharmaceutical composition selected by a skilled practitioner for a particular subject. Regular administration dosing intervals include, for example, once daily, twice daily, once every 2, 3, 4, 5 or 6 days, once or twice weekly, once or twice monthly, and the like.

[0275] The present invention further includes the use of the MASP inhibitory peptides described herein for the manufacture of a medicament, in particular for the manufacture of a medicament for the prevention and / or treatment of a disorder or disease defined herein.

[0276] The present invention further includes a process for producing the peptides of the present invention, their derivatives or analogs, or pharmaceutically acceptable salts or solvates thereof, or conjugates thereof, each as described herein, which comprises the steps set forth in the examples of the present invention.

[0277] In general, the MASP inhibitory peptides of the present invention can be produced synthetically or semi-recombinantly.

[0278] According to a further embodiment, the present invention provides a method for preparing a compound containing a peptide comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which can be isolated and / or purified, or a derivative, prodrug, analog, pharmaceutically acceptable salt or solvate thereof, by using solid phase peptide synthesis.

[0279] According to a further embodiment, the present invention provides a method for preparing a compound containing a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt or solvate or solvate of a salt comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, said method comprising the steps of: 1. Use of 2-chlorotrityl type resin with a loading of 0.2-1.0 mmol / g or Wang type resin with a loading of 0.2-1.0 mmol / g; 2. Load the C-terminal amino acid of the sequence onto the resin; 3. Removal of Fmoc protection with 15-25% piperidine solution in DMF or NMP, 4. Coupling the next amino acid in the sequence using a coupling reagent such as HBTU, HATU or DIC / Oxyma in a stoichiometry of 3-8 equivalents; 5. Repeat steps 3 and 4 until the sequence is complete. 6. Cleavage of the peptide from the solid support using a cleavage cocktail containing TFA and a thiol scavenger. 7. Cyclization of the two cysteines in the sequence under oxidizing conditions (air or I2), 8. Purification of cleaved peptides using reversed-phase HPLC.

[0280] According to a further embodiment, the present invention provides a method for preparing a compound containing a peptide or derivative, prodrug, analogue, pharmaceutically acceptable salt or solvate or solvate of a salt comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), which may be isolated and / or purified, said method comprising the steps of: 1. Use of 2-chlorotrityl type resin with a loading of 0.2-1.0 mmol / g or Wang type resin with a loading of 0.2-1.0 mmol / g; 2. Load the C-terminal amino acid of the sequence onto the resin; 3. Removal of Fmoc protection with 15-25% piperidine solution in DMF or NMP, 4. Coupling the next amino acid in the sequence using a coupling reagent such as HBTU, HATU or DIC / Oxyma in a stoichiometry of 3-8 equivalents; 5. Repeat steps 3 and 4 until the sequence is complete. 6. Cleavage of the peptide from the solid support using a cleavage cocktail containing TFA and a thiol scavenger. 7. Cyclization of the two cysteines in the sequence under oxidizing conditions (air or I2), 8. Purification of cleaved peptides using reversed-phase HPLC 9. Conversion to HCl salt.

[0281] At least one peptide, derivative or analogue defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex defined herein, may also be used as a biochemical agent in a biochemical assay (e.g., in a diagnostic assay for measuring responsiveness to a MASP inhibitor, or in any biochemical assay based on MASP inhibitor binding).

[0282] The present invention also includes polynucleic acids comprising sequences encoding MASP inhibitory peptides according to the present invention, as well as vectors comprising polynucleic acids comprising sequences encoding MASP inhibitory peptides according to the present invention.

[0283] The present invention will be further illustrated by the following examples relating to specific embodiments of the present invention. Unless otherwise specified, the examples were carried out using standard techniques well known and routine to those skilled in the art. The following examples are for illustrative purposes only and are not intended to be completely definitive regarding the conditions or scope of the present invention. Therefore, they should not be interpreted as limiting the scope of the present invention in any way. [Example]

[0284] Example List of abbreviations used in the experimental section: Å Angstrom aq. aqueous, aqueous solution bar unit of pressure BPR back pressure suppression device conc concentration d Double line (NMR) dd doublet doublet (NMR) DCM dichloromethane DEA Diethylamine DIPEA N,N-Diisopropylethylamine (Huenig's base) DMAP 4-dimethylaminopyridine DMF N,N-dimethylformamide DMSO dimethyl sulfoxide dt triplet doublet (NMR) EA Ethyl acetate ee enantiomeric excess ent enantiomer eq Equivalent(s) equiv equivalent (ion chromatography) ESI electrospray ionization (mass spectrometry) Fmoc-OSu 1-{[(9H-fluoren-9-ylmethoxy)carbonyl]oxy}pyrrolidine-2,5-dione GC-MS Gas chromatography coupled to mass spectrometry h time(s) HATU O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium-hexafluorophosphate HPLC High Pressure Liquid Chromatography IC ion chromatography ID Inner Diameter L liters LC-MS Liquid chromatography coupled to mass spectrometry LiHMDS Lithium bis(trimethylsilyl)amide lit. literature m multiplet (NMR) MALDI matrix-assisted laser desorption / ionization (mass spectrometry) Me methyl min minutes(s) mm millimeters μm micrometer (micron) M mole MPLC Medium Pressure Liquid Chromatography MS mass spectrometry MTBE tert-butyl methyl ether MTP Microtiter Plate m / z mass-to-charge ratio (mass spectrometry) nm nanometer NMP N-methyl-2-pyrrolidone NMR nuclear magnetic resonance spectroscopy PBS Phosphate-buffered saline PE Petroleum Ether PEG polyethylene glycol pos positive ppm parts per million Pr Propyl Psi pounds per square inch (pressure) q / quart quartet (NMR) qd doublet quartet (NMR) quint sextet (NMR) rac racemic Rf retention rate (TLC) RP Reversed Phase (for liquid chromatography) Rt retention time (chromatography) s Singlet (NMR) s seconds (hours) SPPS Solid Phase Peptide Synthesis t Triplet (NMR) TBTU O(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate tBu tert-butyl TEA Triethylamine THF tetrahydrofuran TLC thin layer chromatography UPLC Ultra High Performance Liquid Chromatography UV ultraviolet light Further abbreviations can be found in Tables 2 and 3.

[0285] Analytical LC-MS method Method 1 MS system: ThermoFisher Scientific LTQ-Orbitrap-XL; HPLC system: Agilent 1200SL; Column: Agilent, POROSHELL 120, 3 x 150 mm, SB-C18 2.7 μm; Eluent A: 1 L water + 0.1% trifluoroacetic acid; Eluent B: 1 L acetonitrile + 0.1% trifluoroacetic acid; Gradient: 0.0 min 2% B → 1.5 min 2% B → 15.5 min 98% B → 18.0 min 98% B; Oven: 40 °C; Flow rate: 0.75 mL / min; UV detection: 210 nm Method 2 MS system: ThermoFisher Scientific LTQ-Orbitrap-XL, HPLC system: Agilent 1200SL, Column: Agilent, POROSHELL 120; 3 x 150 mm, SB-C18 2.7 μm, Eluent A: 1 L water + 0.1% trifluoroacetic acid, Eluent B: 1 L acetonitrile + 0.1% trifluoroacetic acid, Gradient: 0.0 min 5% B → 0.3 min 5% B → 7.0 min 98% B → 10 min 98% B, Oven: 40°C, Flow rate: 0.75 mL / min, UV detection: 210 nm Method 3 MS instrument type: Waters TOF instrument; UPLC instrument type: Waters Acquity I-Class; Column: YMC, TRIART C18, 75 x 1 mm, 3.0 μm x 12 nm; Eluent A: 1 L water + 0.01% formic acid; Eluent B: 1 L acetonitrile + 0.01% formic acid; Gradient: 0.0 min 1% B → 2.0 min 1% B → 8.0 min 95% B → 10.0 min 95% B; Oven: 50°C; Flow rate: 0.63 mL / min; UV detection: 210 nm Method 4 MS instrument type: Waters TOF instrument; UPLC instrument type: Waters Acquity I-CLASS; Column: YMC, TRIART C18, 75 × 1 mm, 3.0 μm, 12 nm; Eluent A: 1 L water + 0.01% formic acid; Eluent B: 1 L acetonitrile + 0.01% formic acid; Gradient: 0.0 min 1% B → 1.0 min 1% B → 15.0 min 50% B → 18.0 min 95% B; Oven: 50 °C; Flow rate: 0.63 mL / min; UV detection: 210 nm Method 5 MS instrument type: Waters TOF instrument, UPLC instrument type: Waters Acquity I-CLASS, Column: Waters, HSST3, 2.1 x 50 mm, C18 1.8 μm, Eluent A: 1 L water + 0.01% formic acid, Eluent B: 1 L acetonitrile + 0.01% formic acid, Gradient: 0.0 min 2% B → 0.5 min 2% B → 7.5 min 95% B → 10.0 min 95% B, Oven: 50°C, Flow rate: 1.00 mL / min, UV detection: 210 nm Method 6 MS system: Waters Synapt G2S; UPLC system: Waters Acquity I-CLASS; Column: Waters, BEH300, 2.1 x 150 mm, C18 1.7 μm; Eluent A: 1 L water + 0.01% formic acid; Eluent B: 1 L acetonitrile + 0.01% formic acid; Gradient: 0.0 min 2% B → 1.5 min 2% B → 8.5 min 95% B → 10.0 min 95% B; Oven: 50 °C; Flow rate: 0.50 mL / min; UV detection: 220 nm Method 7 MS instrument type: Agilent 6410 Triple Quad; HPLC instrument type: Agilent 1200; Column: Gemini-NX C18 5 μm 110 Å 150 × 4.6 mm; Eluent A: 0.1% TFA in H2O; Eluent B: 0.1% TFA in ACN; Gradient: 0.0 min 20% B → 20 min 50% B → 20.1 min 90% B → 23 min 90% B; Oven temperature: 50 °C; Flow rate: 1.0 mL / min; UV detection: 220 nm Method 8 MS instrument type: Agilent 6410 Triple Quad; HPLC instrument type: Agilent 1200; Column: Discovery BIO Wide Pore C18 5 μm 300 Å × 4.6 mm; Eluent A: 0.1% TFA in H2O; Eluent B: 0.1% TFA in ACN; Gradient: 0.0 min 10% B → 20 min 80% B → 20.1 min 90% B → 23 min 90% B; Oven temperature: 50 °C; Flow rate: 1.0 mL / min; UV detection: 220 nm Method 9 Instrument: Waters ACQUITY SQD UPLC System; Column: Waters Acquity UPLC HSS T3 1.8 μm 50 × 1 mm; Eluent A: 1 L water + 0.25 mL 99% formic acid, Eluent B: 1 L acetonitrile + 0.25 mL 99% formic acid; Gradient: 0.0 min 90% A → 1.2 min 5% A → 2.0 min 5% A; Oven: 50 °C; Flow rate: 0.40 mL / min; UV detection: 210 nm Method 10 MS instrument: Thermo Scientific FT-MS; UHPLC instrument: Thermo Scientific UltiMate 3000; Column: Waters, HSST3, 2.1 × 75 mm, C18 1.8 μm; Eluent A: 1 L water + 0.01% formic acid; Eluent B: 1 L acetonitrile + 0.01% formic acid; Gradient: 0.0 min 10% B → 2.5 min 95% B; Oven: 50 °C; Flow rate: 0.90 mL / min; UV detection: 210 nm / optimal integration path 210–300 nm Method 11 MS instrument: Agilent MS Quad 6150; HPLC: Agilent 1290; Column: Waters Acquity UPLC HSS T3 1.8 μm 50 × 2.1 mm; Eluent A: 1 L water + 0.25 mL formic acid, Eluent B: 1 L acetonitrile + 0.25 mL formic acid; Gradient: 0.0 min 90% A → 0.3 min 90% A → 1.7 min 5% A → 3.0 min 5% A. Oven: 50 °C; Flow rate: 1.20 mL / min; UV detection: 205–305 nm.

[0286] Method 12 Apparatus: Waters ACQUITY SQD UPLC System; Column: Waters Acquity UPLC HSST3 1.8 μm 50x1 mm; Eluent A: 1 L water + 0.25 mL 99% formic acid, Eluent B: 1 L acetonitrile + 0.25 mL 99% formic acid; Gradient: 0.0 min 95% A → 6.0 min 5% A → 7.5 min 5% A. Oven: 50 °C; Flow rate: 0.35 mL / min; UV detection: 210 nm.

[0287] Method 13 Equipment: Waters Single Quad MS System; Instrument Waters UPLC Acquity; Column: Waters BEH 18 1.7 μm 50 × 2.1 mm; Eluent A: 1 L water + 1.0 mL (25% ammonia) / L, Eluent B: 1 L acetonitrile; Gradient: 0.0 min 92% A → 0.1 min 92% A → 1.8 min 5% A → 3.5 min 5% A; Oven: 50 °C; Flow rate: 0.45 mL / min; UV detection: 210 nm.

[0288] Method 14 MS system: Waters TOF instrument; UPLC system: Waters Acquity I-CLASS; Column: Waters Acquity UPLC Peptide BEH C18 300 Å, 1.7 μm 150 × 2.1 mm; Eluent A: 1 L water + 0.100 ml 99% formic acid, Eluent B: 1 L acetonitrile + 0.100 ml 99% formic acid; Gradient: 0.0 min 90% A → 0.25 min 90% A → 8.0 min 45% A → 10.0 min 2% → 12.0 min 2% A. Oven: 50 °C; Flow rate: 0.475 ml / min; UV detection: 210 nm.

[0289] Method 15 MS instrument type: Agilent G6110A; HPLC instrument type: Agilent 1200 Series LC; UV DAD; column: Chromolith Flash RP-18e 25 × 2.0 mm; mobile phase A: 0.0375% TFA (v / v) in water; mobile phase B: 0.01875% TFA (v / v) in acetonitrile; gradient: 0.01 min 5% B → 0.80 min 95% B → 1.20 min 95% B → 1.21 min 5% B → 1.5 min 5% B; flow rate: 1.50 mL / min; oven temperature: 50 °C; UV detection: 220 nm and 254 nm.

[0290] MALDI method Accurate mass measurements were performed on selected peptides using matrix-assisted laser desorption / ionization (MALDI) mass spectrometry on a Bruker autoflex maX LRF MALDI MS Time-of-Flight (ToF-MS) system. Samples were prepared on a Bruker MALDI target plate using α-cyano-4-hydroxycinnamic acid (CAS 28166-41-8) as the matrix. A solution of 0.1–1.0 mg of sample peptide in 1.0 mL of acetonitrile-water (50 / 50 or 30 / 70) and a stock solution of matrix (10 mg / mL) in 50% acetonitrile in water containing 0.05% trifluoroacetic acid were prepared. 1.0 μL of each solution was placed on the MALDI target plate and allowed to dry. The sample was then ready for analysis. Recommended sample preparation for the MALDI target plate can be found in the documentation provided by Bruker.

[0291] Analytical ion chromatography Method: IC-Quantitative Quantitative determination of cations and anions using external standards; Instrument: Thermo Scientific ICS 5000+; Capillary IC Columns: IonPac AS 11-HC and IonPac CS16; Eluent: Gradient eluent [H]+ [OH]-; Detector: Conductivity detection; Routine anions available: acetate, bromide, citrate, chloride, fluoride, formate, lactate, mesylate, phosphate, sulfate, tartrate, trifluoroacetate; Routine cations available: ammonium, barium, calcium, potassium, lithium, sodium, magnesium, choline.

[0292] analytical gas chromatography mass spectrometry GC-MS method 1 Instrument: Thermo Scientific DSQII, Thermo Scientific Trace GC Ultra; Column: Restek RTX-35MS, 15 m x 200 μm x 0.33 μm; Constant flow with helium: 1.20 mL / min; Oven: 60 °C; Inlet: 220 °C; Gradient: 60 °C, 30 °C / min → 300 °C (hold 3.33 min).

[0293] NMR of selected compounds 1 H-NMR data is 1 The peaks are listed in the form of a H-NMR peak list, where for each signal peak, a δ value in ppm is given, followed by the signal intensity reported in parentheses. δ value-signal intensity pairs from different peaks are separated by commas. Thus, the peak list is written in the general format: δ1(intensity 1), δ2(intensity 2), ., δi(intensity i), ., δn(intensity n).

[0294] The intensity of a sharp signal correlates with its height (cm) in the printed NMR spectrum. When compared to other signals, this data can be correlated with the actual ratio of signal intensities. For broad signals, two or more peaks, or signal centers, are shown along with their relative intensities compared to the most intense signal displayed in the spectrum. 1 The H-NMR peak list is a classic 1It is similar to a H-NMR readout and therefore typically includes all peaks listed in a classical NMR interpretation. Furthermore, similar to a classical H-NMR printout, the peak list can show solvent signals, signals originating from stereoisomers of a particular target compound, impurity peaks, C satellite peaks, and / or spinning sidebands. Stereoisomeric and / or impurity peaks typically exhibit lower intensities (e.g., >90% purity) compared to the peaks of the target compound. Such stereoisomers and / or impurities are typical of a particular manufacturing process, and thus, their peaks can be useful for identifying manufacturing process replicas based on "by-product fingerprints." Experts who calculate the peaks of the target compound by known methods (MestReC, ACD simulations, or the use of empirically evaluated expectation values) can optionally use additional intensity filters to separate the peaks of representative compounds, as needed. Such operations are similar to peak picking in classical H-NMR interpretation. A detailed description of reporting NMR data in the form of peak lists can be found in the publication "Citation of NMR Peaklist Data within Patent Applications" (see http: / / www.researchdisclosure.com / searching-disclosures, Research Disclosure Database Number 605005, 2014, 01 Aug 2014). In peak picking routines, the parameter "minimum height" can be adjusted between 1% and 4%, as described in Research Disclosure Database Number 605005. However, depending on the chemical structure and / or concentration of the measured compounds, it is reasonable to set the parameter "minimum height" < 1%.

[0295] Preparation Examples General synthesis method Solid-phase peptide synthesis (SPPS) was performed using an automated peptide synthesizer or manually. Peptide synthesis was typically performed at a scale ranging from 0.1 to 1.0 mmol. When peptide synthesis was performed manually, the general procedures C, D, and E described below were used. Automated peptide synthesis was performed on a Symphony X peptide synthesizer (Gyros Protein Technologies).

[0296] Fmoc-protected amino acids (or intermediate amino acids used to prepare Fmoc amino acids) were purchased from Novabiochem, Bachem, Iris Biotech, Sigma-Aldrich, Alfa, Enamine, Amatek, Anichem, ACBR, ABC Laboratory, AP Bioscience, Combiblocks, ArZa Bioscience, Ark Pharm, Acroteinchem, Apollo Scientific, Biofine, Broadpharm, VWR, or Gyros Protein Technologies (fluorenylmethoxycarbonyl = Fmoc), GL Biochem (Shanghai) Ltd, Chengdu Aminop Pharmaceutical Technology Ltd, Suzhou Highfine Biotech Co. Ltd, WuXi AppTec, or supplied by other Chinese vendors. Some special Fmoc-protected amino acids were synthesized in-house, and these synthetic methods are described herein. Some of the internally synthesized Fmoc amino acids are also commercially available. In cases where Fmoc-protected amino acids were not commercially available, but Boc-protected (tert-butyloxycarbonyl = Boc) unnatural amino acids were commercially available, Fmoc-protected amino acids were prepared from the Boc-protected amino acids by deprotection and reprotection using methods commonly used in the art. CAS numbers for commercially available unnatural amino acids used in the synthesis of the peptides of the invention are, in most cases, included in Table 5. In cases where racemic amino acids (Fmoc or Boc) are purchased, those skilled in the art should recognize that the enantiomers can be separated using chiral chromatography, which has been done in practice to obtain enantiomerically pure amino acids prior to peptide synthesis.

[0297] The following unnatural amino acids have been used in preparing the peptides of the present invention. The Fmoc or Boc protected amino acids were either obtained through commercial sources (CAS numbers available) or synthesized internally by the methods described herein. Table 5 shows the CAS numbers of the chemical groups / amino acids used in peptide synthesis (right column) and the corresponding chemical groups / amino acids present in the peptides (left column).

[0298] Table 5: Availability of unnatural amino acids and chemical groups of the invention [Table 5] TIFF0007667091000027.tif254169TIFF0007667091000028.tif253166TIFF000 7667091000029.tif254163TIFF0007667091000030.tif254168TIFF00076670910 00031.tif254163TIFF0007667091000032.tif255169TIFF0007667091000033.t if254169TIFF0007667091000034.tif255166TIFF0007667091000035.tif224167

[0299] Solid-phase resins were purchased from Novabiochem, Bachem, Iris Biotech, Pcas Biomatrix, GL Biochem (Shanghai) Ltd, CEM, or Protein Technologies. Resin loadings ranged from 0.3 to 1.0 mmol / g. Peptides were synthesized on 2-chlorotrityl resin, Wang resin, or Rink amide-type resin, depending on the desired C-terminus. Cleavage of the fluorenylmethoxycarbonyl (Fmoc) protecting group was achieved using 20% ​​piperidine in dimethylformamide at room temperature. Each Fmoc cleavage step was performed twice. Amino acids were coupled on an automated synthesizer (Symphony X) using 8 equivalents of Fmoc-amino acid, 8 equivalents of DIC (diisopropylcarbodiimide) (0.5 M in DMF), and 8 equivalents of Oxyma (ethyl cyanohydroxyiminoacetate) (0.5 M in DMF). Amino acid coupling was performed at room temperature under a nitrogen atmosphere when using the Symphony X. When expensive or self-prepared fmoc-amino acids were used, couplings were performed manually using 3 equivalents of Fmoc-amino acid with 3 equivalents of DIC (0.5 M in DMF) and 3 equivalents of Oxyma (0.5 M in DMF). Each amino acid coupling step was performed twice (double coupling). Alternatively, peptides were prepared manually by SPPS using 3 equivalents of Fmoc-amino acid, 2.85 equivalents of HBTU (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, benzotriazole tetramethyluronium hexafluorophosphate) (0.5 M in DMF), and 6 equivalents of DIPEA (0.5 M in DMF). The coupling reaction was monitored using the ninhydrin test.

[0300] Peptides were fully deprotected using trifluoroacetic acid (TFA) / thioanisole (TA) / 1,2-ethanedithiol (EDT) (90:7:3) or 92.5% TFA / 2.5% EDT / 2.5% TIS (triisopropylsilane) / 2.5% HO. For peptides containing methionine, the peptide was treated with a solution of 1.6% EDT and 1.2% trimethylsilyl bromide in TFA for 2 h at room temperature to reduce the oxidized methionine.

[0301] Disulfide Cyclization: Disulfide bridges were formed by shaking the peptide overnight in 0.1 M ammonium bicarbonate buffer (pH 7.83) at a concentration of 0.5 mg / mL. The solution was then lyophilized. Alternatively, disulfide bridges were formed by shaking the peptide overnight in solid ammonium bicarbonate buffer at a concentration of 1–3 mg / mL in a mixture of acetonitrile / water (often 3:7) adjusted to pH 9.0. Alternatively, disulfide bridges were prepared by oxidation with iodine (I2) (0.1 M in MeOH) at a concentration of 1–1.3 mg / mL in acetonitrile / water (1:1) for 2 min at 20 °C, followed by treatment with sodium thiosulfate (0.1 M in water), followed by lyophilization.

[0302] Optional acetylation: N-terminal acetylation was carried out using 10 equivalents of acetic anhydride and 2.5 equivalents of DIPEA in DMF (2 mL) by shaking the suspension on an orbital shaker at room temperature for 1 hour. The solvent was removed, and the resin was washed with DMF (5x) and DCM (5x). This procedure was then repeated again. Alternatively, N-terminal acetylation was carried out using 10 mL of a capping solution consisting of acetic anhydride / N-methylmorpholine (NMM) / DMF (10:5:85) by shaking the suspension on an orbital shaker at room temperature for 30 minutes.

[0303] Peptide cleavage: A cleavage cocktail containing TFA / EDT / thioanisole (90:3:7) was prepared. The cleavage cocktail (2 mL) was added to the peptide-containing resin, and the suspension was shaken on an orbital shaker for 2.5 h. Cold ether (-20 °C) was added to precipitate the peptide. The resulting solution was centrifuged (Sigma 2-16KL) under nitrogen, and the solid obtained after decantation was washed three more times with cold ether by centrifugation and decantation. The resulting solid was purified by preparative HPLC.

[0304] Alternatively, a cleavage cocktail containing TFA / EDT / TIS / HO (92.5:2.5:2.5:2.5) was prepared. 6 mL of the cleavage cocktail (0.3 mmol scale) was added to the peptide-containing resin, and the suspension was shaken on an orbital shaker for 2.5 h. The peptide was precipitated by adding cold t-butyl methyl ether (-20 °C). The resulting solution was centrifuged at 3000 rpm for 3 min, and the solid obtained after decantation was washed three more times (20 mL × 3) with cold t-butyl methyl ether by centrifugation and decantation. The resulting crude peptide was dried in vacuo for 2 h and then purified by preparative HPLC.

[0305] Preparative HPLC: An Agilent 1260 Prep reverse-phase HPLC or a Knauer AZURA Prep reverse-phase HPLC was used for purification. Columns were selected based on the results of column screening. Peptides were dissolved in 10-30% ACN / water (typically the starting point of the gradient). Both water and acetonitrile contained 0.1% TFA. A flow rate of 20 mL / min was typically used, ranging from 10-30% ACN / water to 85-90% ACN / water. Fractions were analyzed by HPLC (Agilent 1260 Infinity) using a Chromolith Speedrod column (5-95% ACN / water, 8 min gradient) and by one or more of the following LC-MS methods: Method 1, Method 2, Method 3, Method 4, Method 5, and Method 6.

[0306] Alternatively, a Gilson GX-281 Prep reverse-phase HPLC was used for purification. The column was selected based on the results of a column screen. The peptide was dissolved in 10-30% ACN / water (typically the starting point of the gradient). The water contained 0.075% TFA. Typically, a Luna column (25x200mm, C18 10µm, 110Å) or a Gemini column (30x150mm, C18 5µm, 110Å) was used. The preparative HPLC conditions were: flow rate 20mL / min, 10-30% ACN / water to 85-90% ACN / water, wavelength 214 / 254nm, oven temperature 30°C. o C. Fractions were analyzed by high performance liquid chromatography (Agilent 1260 Infinity) using Method W1 or Method 7. Peptides were then analyzed by one or more of the following methods: Method 1, Method 2, Method 3, Method 4, Method 5, Method 6.

[0307] Disulfide mimetics (in which the -SS-disulfide bond is replaced by -CH2-S-, -S-CH2-, -CH2-CH2-, -S-(CH2)2-, -(CH2)2-S-, or -CH2-S-CH2-) can be made according to the methods described in the following references in combination with the methods described herein: (1) Hong-Kui Cui, Ye Guo, Yao He, Feng-Liang Wang, Hao-Nan Chang, Yu-Jia Wang, Fang-Ming Wu, Chang-Lin Tian, ​​Lei Liu, Angew. Chem. Int. Ed. 2013, 52, 9558-9562; (2) Ye Guo, De-Meng Sun, Feng-Liang Wang, Yao He, Lei Liu, Chang-Lin Tian, ​​Angew.Chem.Int.Ed.2015, 54, 14276-14281; (3) Yang Xu, Tao Wang, Chao-Jian Guan, Yi-Ming Li, Lei Liu, Jing Shi, Donald Bierer, Tetrahedron Letters 2017, 58, 1677-1680; (4) Tao Wang, Yi-Fu Kong, Yang Xu, Jian Fan, Hua-Jian Shi, Ge-Min Fang, Org.Lett.2018, 20, 6074-6078;(6)Jan-Patrick Fischer, Ria Schoenauer, Sylvia Els-Heindl, Donald Bierer, Johannes Koebberling, Bernd Riedl, Annette G. Beck-Sickinger, J Pep Sci.2019;e3147;(7)Dong-Liang Huang, Jing-Si Bai, Meng Wu, Xia Wang, Bernd Riedl, Elisabeth Pook, Carsten Alt, Marion Erny, Yi-Ming Li, Donald Bierer, Jing Shi, Ge-Min Fang, Chem.Commun., 2019, 55, 2821-2824; (8) Shuai-Shuai Sun, Junyou Chen, Rui Zhao, Donald Bierer, Jun Wang, Ge-Min Fang, Yi-Ming Li, Tetrahedron Letters 2019, 60, 1197-1201; (9) CMBKKourra and N. Cramer, Chem.Sci., 2016, 7, 7007-7012. .

[0308] Unless otherwise specified, all peptides of the present invention are TFA salts.

[0309] A general method for automated SPPS of Masp peptides (Method A) The synthesis of ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IPD-NH2 (Example 165) is representative.

[0310] Peptides were synthesized using standard Fmoc chemistry.

[0311] Automated SPPS was performed on a Symphony X peptide synthesizer (Protein Technologies). ChemMatrix Rink Amide resin was typically used at a 0.1 mmol scale (0.5 mmol / g loading). The resin was placed in a reaction vessel and placed on the instrument. The following solutions were prepared and used during the synthesis: 1) Fmoc amino acid: 0.2M (8 equivalents) 2) Activator 1: 0.5M DIC in DMF (7.5 equivalents) 3) Activator 2: 0.5M Oxyma (7.5 equivalents) in DMF 4) Fmoc deprotection: 20% piperidine in DMF 5) Acetic anhydride (10 equivalents) in 2 mL of DMF Double couplings were typically performed for each amino acid. For expensive unnatural Fmoc or Boc amino acids, in-house synthesized Fmoc amino acids, or N-methylated amino acids, the sequence was interrupted and this amino acid was manually coupled (double coupling, but typically using fewer reagents (3–5 equivalents)). After this coupling was complete, the synthesis was typically continued on the synthesizer. When an N-methyl amino acid was added to the sequence, the next amino acid was typically manually coupled as well. All steps were performed at room temperature under nitrogen. Fmoc-Pen and Fmoc-Oic were typically coupled using automated SPPS. Fmoc(N-Me)G was typically coupled manually. Ahx was always coupled manually.

[0312] The resin was swelled and washed with DMF (3 x 3 mL, 10 min). If the resin contained Fmoc, the Fmoc was removed with 20% piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0313] Coupling: Fmoc-Asp(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Asp(Ot-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0314] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0315] Coupling: Fmoc-Pro (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Pro (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0316] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0317] Coupling: Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0318] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0319] Coupling: Fmoc-Pen(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Pen(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0320] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0321] Coupling: Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0322] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0323] Coupling: Fmoc-Oic (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Oic (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0324] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0325] Coupling: Fmoc-Pro (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Pro (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0326] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0327] Coupling: Fmoc-Leu (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Leu (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0328] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0329] Coupling: Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0330] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0331] Coupling: Fmoc-Arg(Pbf) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Arg(Pbf) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0332] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0333] Coupling: Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0334] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0335] Coupling: Fmoc-Cys(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Cys(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0336] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0337] Coupling: Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0338] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0339] Manual coupling during automated SPPS: Fmoc-(N-Me)G (0.2 M in DMF, 5 equiv.) was added to the resin. Activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling proceeded for 2 h with shaking (Thermomixer, rt). The solution was filtered and washed with DMF (1 × 3 mL, 30 s). The coupling step was repeated. Fmoc-(N-Me)G (0.2 M in DMF, 5 equiv.) was added. Activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling proceeded for 2 h with shaking (Thermomixer, rt). The solution was filtered and washed with DMF (6 × 3 mL, 30 s).

[0340] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0341] If additional amino acids were present in the sequence, they were coupled using the steps above.

[0342] Test Cut: When manual couplings were performed, test cleavages were typically performed to monitor the reaction. The test cleavage cocktail was TFA / EDT / thioanisole (90:3:7); shaken at room temperature and 750 rpm on a Thermomixer for 1.5 hours. Analysis was performed by LC-MS using one of the methods described above.

[0343] Fully disconnected: The resin containing the peptide was placed in a syringe, and 3.0 mL of cleavage buffer TFA / EDT / thioanisole (90:3:7) was added to the resin. The mixture was shaken at room temperature for 2.5 hours. The solution was filtered and diluted with cold diethyl ether (-20 oThe peptide was precipitated by adding HCl (C). The solution was centrifuged (3000 rpm) in a Falcon tube (60 mL) under nitrogen atmosphere. The ether was decanted, and the solid residue was washed repeatedly with cold diethyl ether (5 × 10 mL). The solid residue was then dried.

[0344] Disulfide cyclization: The crude peptide was dissolved in 0.1 M ammonium bicarbonate (pH 7.8-8.2) at a concentration of 1 mg / 2 mL. The solution was shaken open to the atmosphere in a round-bottom flask overnight on an orbital shaker. The solution was then lyophilized to yield a white powder.

[0345] Column screening for HPLC purification: Peptides were dissolved in 5% CH3CN and 95% water. Column screening was performed for each peptide to determine which preparative HPLC method to use for purification. The following analytical columns were screened:

[0346] There are two methods for column screening: 1) 5-60%ACN_8min_1mL / min_25°C 2) 30-85%ACN_8min_1mL / min_25°C Usable columns (50mm x ID4,6mm) (can also be used as preparative columns): 1) Aeris C18 (Phenomenex) 2) X-Bridge C18 (Waters) 3) Kinetex C18(Coreshell Material)(Phenomenex) 4) YMC Triart C18 (can be eluted with 100% water) 5) Kinetix Biphenyl(Phenomenex) 6) X-Select C18 (positive charge) 7) Jupiter Proteo C18(Phenomenex) 8) Luna C18 (Phenomenex) For peptides of the present invention, one of the following five preparative columns was used: 1) Column: Phenomenex, Aeris Peptide 5μ XB-C18, AXIA Packed, 21,2x250mm+Cartridge 5μ 2) Column: Phenomenex, Kinetex C18 5μ 21.5x250mm + Cartridge 5μ 3) Column: Phenomenex, Kinetex 5μ Biphenyl 100A, AXIA Packed, 21.2x250mm + Cartridge 5μ 4) Column: YMC Actus Triart Prep. C18 12 nm, S-10 μm 250 x 20 mm + Cartridge 3 μm (10 x 4 mm) 5) Column: Waters, Xbridge Prep.C18.5μ OBD 19x250mm + Cartridge 10μ Once you have selected the columns, use one of the following methods: 1) Method: Gradient 5 to 60% ACN (in water) (0.10% TFA) 2) Method: Gradient 30-85% ACN (in water) (0.10% TFA) 3) Focused gradient based on the results of column screening.

[0347] Flow rate 20mL / min The combined fractions were analyzed using HPLC (5-95 in 8 in, Chromolith SpeedROD & YMC C18 5-95 in 18 min) and one of the LC-MS methods described above.

[0348] For Example 165, the crude peptide was dissolved in CHCN / water (1:1) and purified on a Phenomenex Kinetex column C18 5μ, 21.5 x 250 mm + 5μ cartridge, flow rate: 20 mL / min, method: 5-60% ACN in water (each containing 0.10% TFA) over 48 min. The combined fractions were lyophilized to give 39 mg of Example 165 (95% purity).

[0349] Table 6: Notes on materials and conditions used [Table 6]

[0350] General method for automated SPPS of Masp peptides (Method B) The synthesis of ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+I-((5-azaspiro[2.4]heptane-1-carboxylic acid)-OH (Example 164) is representative.

[0351] Peptides were synthesized using standard Fmoc chemistry.

[0352] When possible, Wang resin preloaded with the first amino acid (e.g., preloaded with Fmoc-Asp(t-Bu)-OH, Fmoc-Pro-OH, etc.) was used, with a typical loading of 0.55–0.7 mmol / g. If the first amino acid was not available, it was added manually.

[0353] If the first amino acid was not available as a preloaded resin, it was added manually and chlorotrityl resin was used for the synthesis.

[0354] Loading of the first amino acid onto the 2-chlorotrityl resin: 2-Chlortrityl resin (500 mg, 0.775 mmol) was swelled in 10 mL of DCM in a 50 mL Falcon tube for 15 min. 5-[(9H-fluoren-9-ylmethoxy)carbonyl]-5-azaspiro[2.4]heptane-1-carboxylic acid (281 mg, 0.775 mmol) was dissolved in DCM with DIEA (6 equiv., 0.81 mL) and the solution was added to the resin. The solution was purged with argon and shaken overnight at room temperature. The mixture was filtered and washed with DMF (3 × 5 mL) and DCM (3 × 5 mL). Methanol (5 mL) was added, and the mixture was shaken for 30 min and then filtered. The resin was washed with DMF (3 × 5 mL) and DCM (3 × 5 mL). Methanol (5 mL) was added again, and the mixture was shaken for 30 min and then filtered. The resin was washed with DMF (3 x 5 mL) and DCM (3 x 5 mL). The loading was determined to be 0.42 mmol / g based on the determination of the loading procedure described below.

[0355] This resin was then used for automated SPPS on a Symphony X synthesizer from Protein Technologies at a 0.1 mmol scale.

[0356] Determining Resin Loading: To determine resin loading, the FMOC protecting group is cleaved from a defined amount of resin, and the concentration of the resulting fluorenyl compound in the supernatant cleavage solution is then measured photometrically at 301 nm, which directly correlates with the amount of amino acid loaded onto the resin.

[0357] 1) Weigh 1-3 mg of resin into a 2 mL Eppendorf tube or similar (note the exact amount).

[0358] 2) Add 1000 μL of a solution of 20% piperidine in DMF.

[0359] 3) The mixture is stirred for 30 minutes to cleave the FMOC group.

[0360] 4) Then, 100 μL of the supernatant solution is transferred to a quartz glass cuvette and diluted with 900 μL of 20% piperidine / DMF. 5) Prepare a blank sample of 1000 μl piperidine / DMF in a second cuvette.

[0361] 6) After determining the blank value from the reference sample, measure the absorbance of the test sample at λ=301 nm with a UV-Vis photometer (Thermo Scientific Evolution 201).

[0362] 7) For greater accuracy, multiple test samples (typically two) can be formed and the arithmetic mean of the measured absorbances is then used for the calculation.

[0363] Resin Loading Calculation: Resin loading in mmol / g L 301 is calculated as follows: Equation 1

[0364] TIFF0007667091000037.tif20135E=Absorbance ε = extinction coefficient at a wavelength of 301 nm (7800 L / mol * cm) m=Amount of resin used (g) V = sample volume (L) D = cuvette layer thickness (cm) VF = dilution factor (=10) Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0365] Subsequent coupling of Fmoc amino acids proceeded as described in Method A.

[0366] The following solutions were prepared and used during the synthesis: 1) Fmoc amino acid: 0.2M (8 equivalents) 2) Activator 1: 0.5M DIC in DMF (7.5 equivalents) 3) Activator 2: 0.5M Oxyma (7.5 equivalents) in DMF 4) Fmoc deprotection: 20% piperidine in DMF 5) Acetic anhydride (10 equivalents) in 2 mL of DMF Double couplings were typically performed for each amino acid. For expensive unnatural Fmoc or Boc amino acids, in-house synthesized Fmoc amino acids, or N-methylated amino acids, the sequence was interrupted and this amino acid was manually coupled (double coupling, but typically using fewer reagents (3–5 equivalents)). After this coupling was complete, the synthesis was typically continued on the synthesizer. When an N-methyl amino acid was added to the sequence, the next amino acid was typically manually coupled as well. All steps were performed at room temperature under nitrogen. Fmoc-Pen and Fmoc-Oic were typically coupled using automated SPPS. Fmoc(N-Me)G was typically coupled manually. Ahx was always coupled manually.

[0367] Fmoc cutting: The resin was swelled and washed with DMF (3 x 3 mL, 10 min). If the resin contained Fmoc, the Fmoc was removed with 20% piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0368] Coupling: Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0369] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0370] Coupling: Fmoc-Pen(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Pen(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0371] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0372] Coupling: Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0373] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0374] Coupling: Fmoc-Oic (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Oic (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0375] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0376] Coupling: Fmoc-Pro (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Pro (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0377] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0378] Coupling: Fmoc-Leu (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Leu (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0379] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0380] Coupling: Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0381] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0382] Coupling: Fmoc-Arg(Pbf) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Arg(Pbf) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0383] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0384] Coupling: Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ser(t-Bu) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0385] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0386] Coupling: Fmoc-Cys(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Cys(Trt) (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0387] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0388] Coupling: Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (1 × 3 mL, 30 seconds). The coupling step was repeated. Fmoc-Ile (4.0 mL) was added. Activator 1 solution (DIC, 1.5 mL) and Activator 2 solution (Oxyma, 1.5 mL) were added and the coupling was allowed to proceed for 2 hours with nitrogen bubbling. The solution was drained and washed with DMF (6 × 3 mL, 30 seconds).

[0389] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0390] Manual coupling during automated SPPS: Fmoc-(N-Me)G (0.2 M in DMF, 5 equiv.) was added to the resin. Activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling proceeded for 2 h with shaking (Thermomixer, rt). The solution was filtered and washed with DMF (1 × 3 mL, 30 s). The coupling step was repeated. Fmoc-(N-Me)G (0.2 M in DMF, 5 equiv.) was added. Activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) were added, and the coupling proceeded for 2 h with shaking (Thermomixer, rt). The solution was filtered and washed with DMF (6 × 3 mL, 30 s).

[0391] Fmoc cutting: The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 s).

[0392] If additional amino acids were present in the sequence, they were coupled using the steps above.

[0393] Test Cut: When manual couplings were performed, test cleavages were typically performed to monitor the reaction. The test cleavage cocktail was TFA / EDT / thioanisole (90:3:7); shaken at room temperature and 750 rpm on a Thermomixer for 1.5 hours. Analysis was performed by LC-MS using one of the methods described above.

[0394] Fully disconnected: The resin containing the peptide was placed in a syringe, and 3.0 mL of cleavage buffer TFA / EDT / thioanisole (90:3:7) was added to the resin. The mixture was shaken at room temperature for 2.5 hours. The solution was filtered and diluted with cold diethyl ether (-20 oThe peptide was precipitated by adding HCl (C). The solution was centrifuged (3000 rpm) in a Falcon tube (60 mL) under nitrogen atmosphere. The ether was decanted, and the solid residue was washed repeatedly with cold diethyl ether (5 × 10 mL). The solid residue was then dried.

[0395] Disulfide Cyclization: The crude peptide was dissolved in 0.1 M ammonium bicarbonate (pH 7.8-8.2) at a concentration of 1 mg / 2 mL. The solution was shaken open to the atmosphere in a round-bottom flask overnight on an orbital shaker. The solution was then lyophilized to yield a white powder.

[0396] Column screening for HPLC purification: Peptides were dissolved in 5% CH3CN and 95% water. Column screening was performed for each peptide to determine which preparative HPLC method to use for purification. The following analytical columns were screened:

[0397] There are two methods for column screening: 1) 5~60%ACN_8 min_1 mL / min_25°C 2) 30-85% ACN_8 min_1 mL / min_25°C Usable columns (50mm x ID 4.6mm) (can also be used as preparative columns): 1) Aeris C18 (Phenomenex) 2) X-Bridge C18 (Waters) 3) Kinetex C18(Coreshell Material)(Phenomenex) 4) YMC Triart C18 (can be eluted with 100% water) 5) Kinetix Biphenyl(Phenomenex) 6) X-Select C18 (positive charge) 7) Jupiter Proteo C18(Phenomenex) 8) Luna C18 (Phenomenex) For peptides of the present invention, one of the following five preparative columns was used: 1) Column: Phenomenex, Aeris Peptide 5μ XB-C18, AXIA Packed, 21,2x250mm+Cartridge5μ 2) Column: Phenomenex, Kinetex C18 5μ 21.5x250mm + Cartridge 5μ 3) Column: Phenomenex, Kinetex 5μ Biphenyl 100A, AXIA Packed, 21.2x250mm + Cartridge 5μ 4) Column: YMC Actus Triart Prep. C18 12 nm, S-10 μm 250 x 20 mm + Cartridge 3 μm (10 x 4 mm) 5) Column: Waters, Xbridge Prep. C18 5μ OBD 19x250mm + Cartridge 10μ Once you have selected the columns, use one of the following methods: 1) Method: Gradient 5-60% ACN (in water) (0.10% TFA) 2) Method: Gradient 30-85% ACN (in water) (0.10% TFA) 3) Focused gradient based on the results of column screening.

[0398] Flow rate 20mL / min The combined fractions were analyzed by high performance liquid chromatography (5-95 in 8 in, Chromolith SpeedROD & YMC C18 5-95 in 18 min) and using one of the LC-MS methods described above.

[0399] For Example 164, the crude peptide was dissolved in CHCN / water (1:1) and purified on a Phenomenex Kinetex column C18 5μ, 21.5 x 250 mm + 5μ cartridge, flow rate: 20 mL / min, 5-60% ACN in water (each containing 0.10% TFA) over 48 min. The combined fractions were lyophilized to give 29.8 mg of Example 164 (98.3% purity).

[0400] Table 7: Notes on materials and conditions used [Table 7]

[0401] General method for manual SPPS of Masp peptides (Method C) The synthesis of the sequence AIC+SRSLP-(Oic)-I-(Pen)+IPN-OH (Example 418) is representative.

[0402] Peptide synthesis: Peptides were synthesized using standard Fmoc chemistry.

[0403] 1) Resin preparation: To chlorotrityl resin (CTC resin) (0.3 mmol, 0.3 g, 1.0 mmol / g) was added Fmoc-Asn(Trt)-OH (0.3 mmol, 0.18 g, 1.0 equiv.) and DIEA (0.21 mL, 1.2 mmol, 4.0 equiv.) in DCM (6.0 mL). The mixture was stirred with N2 at 20 °C for 2 h, then MeOH (0.3 mL) was added and further stirred with N2 for 30 min. The resin was washed with DMF (6.0 mL × 3). Then, 20% piperidine in DMF (6.0 mL) was added and stirred with N2 at 20 °C for 20 min. The mixture was then filtered to obtain the resin. The resin was washed with DMF (6.0 mL × 3) and filtered to obtain the resin.

[0404] 2) Coupling: Fmoc-Pro-OH (0.30 g, 0.9 mmol, 3.0 equiv.), HBTU (0.32 g, 0.85 mmol, 2.85 equiv.), and DIEA (1.80 mmol, 0.32 mL, 6.00 equiv.) in DMF (3.0 mL) were added to the resin and stirred with N2 at 20 °C for 30 min. The resin was then washed with DMF (6.0 mL × 5).

[0405] 3) Deprotection: 20% piperidine in DMF (6.0 ml) was added to the resin, and the mixture was stirred with N2 at 20° C. for 20 minutes.

[0406] 4) Repeat steps 2-3 for all other amino acids: Table 8: Notes on materials and conditions used [Table 8]

[0407] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin (all amino acids except Pro) and chloranil test (Pro), and the resin was washed five times with DMF (5.0 mL).

[0408] Peptide cleavage and purification: 1) The resin was washed with MeOH (6.0 mL × 5) and dried under vacuum to obtain 0.62 g of peptide resin. Then, 6.0 mL of cleavage buffer (92.5% TFA / 2.5% EDT / 2.5% TIS / 2.5% HO) was added to the flask containing the side-chain protected peptide resin at 20 °C, and the mixture was stirred for 2.5 hours.

[0409] 2) The peptide was precipitated with cold tert-butyl methyl ether (50 mL) and centrifuged (3000 rpm for 3 min) to obtain a solid crude product. The crude peptide precipitate was washed twice more with tert-butyl methyl ether (20.0 mL × 3). The crude peptide was dried under vacuum for 2.0 h to obtain 0.4 g of crude peptide.

[0410] 3) The crude product (0.4 g) was dissolved in CH3CN (150 mL) and water (150 mL). Iodine (I2) (0.1 M in MeOH, 1.5 ml) was added at 20 °C until the yellow color persisted. The mixture was then stirred at 20 °C for 2 minutes. Sodium thiosulfate (0.1 M in water, 0.05 mL) was then added dropwise until the yellow color disappeared. The mixture was lyophilized to give a crude powder (0.41 g). 4) The crude peptide was purified by preparative HPLC (conditions: A: 0.075% TFA in water B: CH3CN) and lyophilized to give the desired peptide (Example 418) (108.1 mg, 23.0% yield, 97.3% purity by Method W1; 97.4% purity by Method 7) as a white solid and TFA salt. Purification conditions: The peptide was dissolved in TFA / HO (7:3) and purified on a Luna 25 x 200 mm, C18 10 µm, 110 Å column at a flow rate of 20 mL / min, a gradient of 12-42% for 60 min; retention time = 42 min, and then purified again on a Gemini 150 x 30 mm, C18 5 µm, 110 Å column to reach the desired purity.

[0411] When C-terminal amides were prepared according to this method, the synthesis began with Rink Amide MBHA resin (typically loading about 0.4-0.6 mmol / g, same steps as above).

[0412] General method for manual SPPS of Masp peptides (Method D) When preparing C-terminal amides, synthesis begins with Rink Amide MBHA resin (typically loading approximately 0.4-0.6 mmol / g), and the sequence PIC+SRS-((tBu)A)-PPI-(Pen)+IPD-NH2 (Example 9) is representative.

[0413] When the C-terminal acid was prepared, the synthesis started with 2-chlorotrityl resin and the first Fmoc amino acid was added as described in Method C.

[0414] Peptide synthesis: Peptides were synthesized using standard Fmoc chemistry.

[0415] 1) Resin preparation: Rink Amide MBHA resin (0.66 g, 0.30 mmol, loading = 0.45 mmol / g) in DMF (10.0 ml) was stirred with N2 at 20 °C for 0.5 h. Then, 20% piperidine in DMF (10 ml) was added and stirred with N2 at 20 °C for 20 min. The mixture was then filtered to obtain the resin. The resin was washed with DMF (10 × 5 mL) and filtered to obtain the resin.

[0416] 2) Coupling: Fmoc-Asn(Trt)-OH (0.9 mmol, 0.37 g, 3.0 equiv.), HBTU (0.32 g, 0.85 mmol, 2.95 equiv.), and DIEA (1.80 mmol, 0.32 mL, 6.00 equiv.) in DMF (3.0 mL) were added to the resin and stirred with N at 20 °C for 30 min. The resin was then washed with DMF (10.0 mL × 3).

[0417] 3) Deprotection: 20% piperidine in DMF (10.0 ml) was added to the resin and the mixture was stirred with N2 at 20° C. for 20 minutes.

[0418] 4) Repeat steps 2-3 for all other amino acids.

[0419] Table 9: Notes on materials and conditions used [Table 9]

[0420] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin (all amino acids except Pro) and chloranil test (Pro), and the resin was washed five times with DMF (5.0 mL).

[0421] Peptide cleavage and purification: 1) The resin was washed with MeOH (6.0 mL × 5) and dried under vacuum to obtain 0.66 g of peptide resin. Then, 6.0 mL of cleavage buffer (90% TFA / 5% EDT / 2.5% TIS / 2.5% HO) was added to the flask containing the side-chain protected peptide resin at 20 °C, and the mixture was stirred for 2.5 hours.

[0422] 2) The peptide was precipitated with cold tert-butyl methyl ether (50 mL) and centrifuged (3000 rpm for 3 minutes) to obtain a solid crude product. The crude peptide precipitate was washed three more times with tert-butyl methyl ether (20.0 mL x 3). The crude peptide was dried under vacuum for 2.0 hours to obtain 0.4 g of crude peptide.

[0423] 3) The crude product (0.4 g) was dissolved in CH3CN (15 mL) and water (15 mL) to obtain a concentration of 0.1 mM. NH4HCO3 solution (1 M) was added to adjust the pH to approximately 8-9. The solution was shaken at room temperature for approximately 8 hours. The reaction was monitored by high-performance liquid chromatography. After completion of the reaction, the reaction was quenched with acetic acid and the pH was adjusted to approximately 6. The reaction mixture was then lyophilized, and the resulting solid was purified by reverse-phase HPLC.

[0424] 4) The crude peptide was purified by preparative HPLC (conditions: A: 0.075% TFA in water, B: CH3CN) and lyophilized to give the desired peptide (Example 9) (202.40 mg, 41.4% yield, 93.7% purity by Method W1; 95.1% purity by Method 7) as a white solid and TFA salt. Purification conditions: Peptide dissolved in TFA / HO (7:3); flow rate 20 mL / min; gradient 12-42% over 60 min; retention time = 42 min; purified on a Luna 25 x 200 mm, C18 10 μm, 110 Å column.

[0425] Another general method for manual SPPS of Masp peptides (Method E) The synthesis of the sequence ((2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid)-IC+SRSLP-(Oic)-IC+I-OH (Example 184) is representative.

[0426] Peptide synthesis: Peptides were synthesized using standard Fmoc chemistry.

[0427] The dark balls in the chemical structures below represent solid-phase polymer supports used for solid-phase peptide synthesis (SPPS), e.g., 2-chlorotrityl resin, Rink amide resin, etc. Example 1A (2S)({[(9H-Fluoren-9-yl)methoxy]carbonyl}amino)(oxan-4-yl)acetic acid (single stereoisomer) [ka]

[0428] To (2S)-amino(oxan-4-yl)acetic acid (950 mg, 5.97 mmol) in acetone / water (15 mL / 10 mL) was added sodium bicarbonate (5.01 g, 59.7 mmol) and 1-({[(9H-fluoren-9-yl)methoxy]carbonyl}oxy)pyrrolidine-2,5-dione (2.11 g, 6.27 mmol). The mixture was stirred at room temperature over the weekend. The suspension was treated with water and extracted twice more with MBTE. The aqueous phase was acidified with 1 M aqueous hydrochloric acid and extracted three times more with dichloromethane. The organic phase was washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, and evaporated. The residue was purified by silica gel chromatography (Isolera; 50 g SNAP Ultra; gradient: 50% cyclohexane / 50% ethyl acetate + 1% acetic acid to 20% cyclohexane / 80% ethyl acetate + 1% acetic acid). The appropriate fractions were combined and evaporated. The residue was dissolved in toluene and evaporated two more times, and the residue was then dried under vacuum to give 1.08 g (100% pure, 47% yield) of the target compound.

[0429] LC-MS (Method 9): R t = 0.90 min; MS (ESIpos): m / z = 382 [M+H] + 1 H-NMR (400 MHz, DMSO-d6) δ [ppm]: -0.150 (0.43), -0.008 (4.33), 0.008 (3.74), 0.146 (0.43), 1.280 (1.03), 1.300 (2.26), 1.309 (2.36), 1.331 (2.75), 1.341 (2.92), 1.371 (2.77), 1.401 (2.54), 1.411 (2.32), 1.430 (1.49), 1.464 (6.34), 1.496 (3.49), 1.932 (1.76), 1.942 (2.14), 1.951 (1.93), 1.960 (2.08), 2.327 (0.64), 2.366 (0.64), 2.670 (0.64), 2.710 (0.65), 3.169 (2.11), 3.212 (2.67), 3.237 (6.32), 3.265 (6.90), 3.704 (0.49), 3.715 (0.49), 3.840 (4.66), 3.854 (5.62), 3.867 (4.30), 3.879 (6.28), 3.899 (5.14), 3.918 (3.56), 4.201 (2.11), 4.218 (5.83), 4.237 (9.07), 4.263 (8.42), 4.274 (9.80), 4.293 (5.14), 4.318 (1.05), 7.309 (6.35), 7.328 (15.12), 7.347 (9.86), 7.400 (9.65), 7.419 (15.88), 7.437 (7.02), 7.652 (5.77), 7.674 (5.52), 7.735 (8.21), 7.740 (8.28), 7.754 (7.63), 7.883 (16.00), 7.902 (14.80), 12.609 (0.56). Example 2A

Chem.

[0430] The reaction was carried out under an argon atmosphere. To N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-isoleucine (9.40 g, 26.6 mmol) in 53 mL of dichloromethane, N,N-diisopropylethylamine (19 mL, 110 mmol) and 2-chlorotrityl chloride resin (10.0 g, 13.3 mmol) were added. The reaction mixture was shaken overnight at room temperature. The precipitate was collected by filtration and washed three times with DMF. The recovered resin was shaken in 1:1 dichloromethane / methanol for 30 minutes. The resin was collected by filtration and washed three times with alternating methanol and dichloromethane. The recovered resin was dried under vacuum for the next step.

[0431] Example 3A [ka]

[0432] Example 2A (15.2 g, 6.82 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 15 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, washing three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0433] Example 4A [ka]

[0434] Example 3A (11.4 g, 5.13 mmol) was swollen in 100 mL of DMF at room temperature for 5 minutes. A mixture of N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-S-(triphenylmethyl)-L-cysteine ​​(6.01 g, 10.3 mmol), DIC (1.5 mL, 10 mmol), and Oxyma (1.42 g, 10.0 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken at room temperature for 2 hours. The resin was collected by filtration and washed three times with DMF. The coupling process was repeated overnight. The resin was collected by filtration and washed three times with 150 mL of DMF, followed by three more times with alternating 150 mL of methanol and 150 mL of dichloromethane. The recovered resin was dried under vacuum for the next step.

[0435] Example 5A [ka]

[0436] Example 4A (18.9 g, 8.51 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 15 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0437] Example 6A [ka]

[0438] Example 5A (16.3 g, 7.32 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-isoleucine (5.18 g, 14.6 mmol), DIC (2.2 mL, 14 mmol), and Oxyma (2.03 g, 14.3 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken at room temperature for 2 hours. The resin was collected by filtration and washed three times with DMF. The coupling process was repeated overnight. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more times with alternating 200 mL of methanol and 200 mL of dichloromethane. The recovered resin was dried under vacuum for the next step. Example 7A [ka]

[0439] Example 6A (20.0 g, 8.98 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 15 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0440] Example 8A [ka]

[0441] Example 7A (17.8 g, 7.99 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of (2S,3aS,7aS)-1-{[(9H-fluoren-9-yl)methoxy]carbonyl}octahydro-1H-indole-2-carboxylic acid (6.25 g, 16.0 mmol), DIC (2.4 mL, 16 mmol), and Oxyma (2.21 g, 15.6 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken at room temperature for 2 hours. The resin was collected by filtration and washed three times with DMF. The coupling process was repeated over the weekend. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more times with alternating 200 mL of methanol and 200 mL of dichloromethane. The recovered resin was dried under vacuum for the next step.

[0442] Example 9A [ka]

[0443] Example 8A (21.5 g, 9.68 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF, then three more times with 200 mL of methanol and 200 mL of dichloromethane, alternating. 200 mL of DMF / piperidine (4:1) was added to the collected resin and shaken at room temperature for 1 hour. The resin was collected by filtration and washed three times with DMF, then three more times with 200 mL of methanol and 200 mL of dichloromethane, alternating. The collected resin was dried under vacuum for the next step.

[0444] Example 10A [ka]

[0445] Example 9A (18.1 g, 8.13 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of 1-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-proline (11.0 g, 32.5 mmol), DIC (4.9 mL, 32 mmol), and Oxyma (4.51 g, 31.7 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken at room temperature over the weekend. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more washes with 200 mL of methanol and 200 mL of dichloromethane, alternating between each other. The collected resin was dried under vacuum for the next step.

[0446] Example 11A [ka]

[0447] Example 10A (20.4 g, 9.16 mmol) in 150 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0448] Example 12A [ka]

[0449] Example 11A (18.2 g, 8.18 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-leucine (11.6 g, 32.7 mmol), DIC (4.9 mL, 32 mmol), and Oxyma (4.53 g, 31.9 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken overnight at room temperature. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0450] Example 13A [ka]

[0451] Example 12A (20.9 g, 9.42 mmol) in 150 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0452] Example 14A [ka]

[0453] Example 13A (18.0 g, 8.11 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of O-tert-butyl-N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-serine (12.4 g, 32.4 mmol), DIC (4.9 mL, 32 mmol), and Oxyma (4.49 g, 31.6 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken overnight at room temperature. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0454] Example 15A [ka]

[0455] Example 14A (21.4 g, 9.64 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0456] Example 16A [ka]

[0457] Example 15A (18.9 g, 8.51 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. 2 -{[(9H-fluoren-9-yl)methoxy]carbonyl}-N 5A mixture of -[N-(2,2,4,6,7-pentamethyl-2,3-dihydro-1-benzofuran-5-sulfonyl)carbamimidoyl]-L-ornithine (22.1 g, 34.0 mmol), DIC (5.1 mL, 33 mmol), and Oxyma (4.72 g, 33.2 mmol) was added, and the reaction mixture was shaken overnight at room temperature. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more washes with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0458] Example 17A [ka]

[0459] Example 16A (23.8 g, 10.7 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0460] Example 18A [ka]

[0461] Example 17A (22.1 g, 9.95 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of O-tert-butyl-N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-serine (15.3 g, 39.8 mmol), DIC (6.0 mL, 39 mmol), and Oxyma (5.51 g, 38.8 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken overnight at room temperature. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0462] Example 19A [ka]

[0463] Example 18A (24.7 g, 11.1 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0464] Example 20A [ka]

[0465] Example 19A (22.2 g, 9.99 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-S-(triphenylmethyl)-L-cysteine ​​(23.4 g, 40.0 mmol), DIC (6.0 mL, 39 mmol), and Oxyma (5.54 g, 39.0 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken overnight at room temperature. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0466] Example 21A [ka]

[0467] Example 20A (24.5 g, 11.0 mmol) in 200 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0468] Example 22A [ka]

[0469] Example 21A (22.3 g, 10.0 mmol) was swollen in 150 mL of DMF at room temperature for 5 minutes. A mixture of N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-isoleucine (14.2 g, 40.1 mmol), DIC (6.0 mL, 39 mmol), and Oxyma (5.55 g, 39.0 mmol) in 50 mL of DMF was added, and the reaction mixture was shaken at room temperature over the weekend. The resin was collected by filtration and washed three times with 200 mL of DMF, followed by three more washes with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum for the next step.

[0470] Example 23A [ka]

[0471] Example 22A (1.00 g, 250 μmol) in 7.5 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating portions of methanol and dichloromethane. The collected resin was dried under vacuum for the next step.

[0472] Example 24A [ka]

[0473] Example 23A (1.00 g, 250 μmol) was swollen in 5 mL of DMF at room temperature for 5 minutes. A mixture of (2S)-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino(oxan-4-yl)acetic acid (381 mg, 1.00 mmol), DIC (150 μL, 980 μmol), and Oxyma (139 mg, 975 μmol) in 1 mL of DMF was added, and the reaction mixture was shaken at room temperature over the weekend. The resin was collected by filtration and washed three times with DMF, followed by three additional washes with alternating methanol and dichloromethane. The recovered resin was dried under vacuum for the next step.

[0474] Example 25A [ka]

[0475] Example 24A (1.00 g, 250 μmol) in 7.5 mL of DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed three times with DMF. Both steps were repeated, and the collected resin was washed three times with alternating portions of methanol and dichloromethane. The collected resin was dried under vacuum for the next step.

[0476] Example 26A (2S,3S)-2-{[(2R)-2-{[(2S,3S)-2-{[(2S,3aS,7aS)-1-{(2S)-1-[(2S,5S,8S,11S,14R,17S,20S)-20-amino-17-[(2S)-butan-2-yl]-8-(3-carbamimidamidopropyl)-5,11-bis(hydroxymethyl)-2-(2-methylpropyl)-20-(oxan-4- (yl)-4,7,10,13,16,19-hexaoxa-14-(sulfanylmethyl)-3,6,9,12,15,18-hexaazaicosan-1-oyl]pyrrolidine-2-carbonyl}octahydro-1H-indole-2-carbonyl]amino}-3-methylpentanoyl]amino}-3-sulfanylpropanoyl]amino}-3-methylpentanoic acid (single stereoisomer) [ka]

[0477] To Example 25A (1.00 g, 250 μmol) was added a cleavage cocktail of TFA / EDT / thioanisole (90:3:7), and the mixture was shaken at room temperature for 2 hours. The filtrate was collected, and the resin was washed with dichloromethane. The combined filtrate was evaporated. The residue was stirred with diethyl ether, and the solid was collected by filtration. The solid was washed several times with diethyl ether and dried under vacuum for the next step.

[0478] Example 184 ((2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid)-IC + SRSLP-(Oic)-IC + I-OH (Example 184) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-18-({N-[(2S)-2-amino-2-(oxan-4-yl)acetyl]-L-isoleucyl}amino)-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-6 -(2-Methylpropyl)-5,8,11,14,17,25,28,35-octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucine (single stereoisomer) [ka]

[0479] To Example 26A (425 mg, 304 μmol) was added 850 mL of 0.1 M aqueous ammonium bicarbonate solution (pH 7.86). Air was bubbled through the reaction mixture for 5 minutes. The reaction mixture was stirred overnight at room temperature in an open flask. The solvent was lyophilized to give 580 mg of a white solid. Purification using reverse-phase HPLC (Method B) as described above gave three fractions of the desired Example 184: 7.4 mg (>99%), 24.5 mg (98%), and 15 mg (95%).

[0480] General Method F: Conversion of Peptide TFA Salts to HCl Salts The synthesis of AIC+SRS-((tBu)A)-PPI-((N-Me)C)+IPD-NH2 (HCl salt) (Example 30) is representative.

[0481] Automated Ion Exchange Station Procedure: Hirschmann peristaltic pump (Rotarus volume 50), tubing: Tygon 2001 (ID 0.64 mm) Settings: Wash with H2O: Runtime 1200 s; 80 min -1 1 cycle (35ml capacity) Sample circulation with peptide: runtime 1200 s; 80 min -1 1 cycle (70ml capacity) Wash with HO (or %ACN in HO): runtime 1200 s; 80 min -1 1 cycle (35ml capacity) Amberlite IRA 410 (HCl form) was used. 1500 mg of resin was placed in two filter cartridges and washed with deionized water (10 times).

[0482] The peptide dissolved in 3 ml of 5% ACN / H2O solution was loaded onto the column and circulated 10 times. The column was washed with water, and the solution was collected in a Falcon tube and lyophilized.

[0483] 72.83 mg of the desired peptide was obtained as the HCl salt: LC-MS (>99%); ion chromatography analysis: 3.7 wt% Cl − (1.57 eq Cl − ), <1 wt% TFA.

[0484] The ion exchange process can also be carried out using the following protocol: Amberlite IRA 410 resin (HCl form) (1-2 g) was placed in a 10 mL fritted syringe (100 mg of peptide requires 1 g of IRA 410 resin). 1) Wash the resin with water (10 times x 3 mL) 2) Wash the resin with 5% ACN in water (1 x 3 mL) 3) The peptide was dissolved in 5% ACN in water. 4) Add the peptide to the syringe and circulate the solution through the column 10-20 times. Collect the eluate in a Falcon tube.

[0485] 5) Wash the resin with 5% ACN in water (10 times 3 mL); add this solution to the solution in the Falkon-tube 6) Freeze-dry the combined solution.

[0486] General Method FA: Conversion of Peptide TFA Salts to Other Salts Other salt forms: The chloride counterion can be exchanged for other counterions by repeatedly passing a solution of the desired salt form (e.g., sodium acetate) through the column, then repeatedly washing the column with water. The peptide is then loaded and the procedure described above is followed. In this way, acetate, tartrate, citrate, and lactate salts of peptides have been prepared. The following three examples are representative: Example 249 Sequence: ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH (acetate) (Example 249) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl) propyl)-5,8,11,14,17,25,28,35-octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolinamide, acetate IRA410 chloride resin (12 g) was placed in an empty 20 mL Biotage column, and the resin was washed (10 times) with a 5% solution of ACN-water. The cartridge was then installed in an automated ion exchange station. The resin was washed with 20 column volumes of 1 M NaOH solution (2 x 1800 s, 80 min). -1) The column was washed with water until the pH of the eluent was below pH 9 (10 column volumes), then the column was eluted with 1 M acetic acid solution (10 column volumes) and washed with water until the pH of the eluent was above pH 5 (approximately 10 column volumes). The peptide (N-Me)GIC+SRSLP-(Oic)-I-Pen+IP-NH2 (TFA salt) (1200 mg) was loaded onto the column in 12 mL of 5% ACN / water and cycled through the column 10 times according to General Method F. The resin was then washed 10 times with 5% ACN / water solution, and the eluate was collected in a 50 mL Falcon tube. The parameter settings used for this scale were as follows: Wash with H2O: Runtime 3085s; 80min; 1 cycle (90ml volume) Sample circulation with peptide: Runtime 3085 s; 80 min; 1 cycle (180 mL volume) Wash with HO (or %ACN in HO): Runtime 3085 s; 80 min; 1 cycle (90 ml volume) The combined eluates were lyophilized to give 1120 mg (100% purity, 97% yield) of the target acetate salt.

[0487] Ion chromatography (chloride content): <1% chloride Ion chromatography (TFA content): <1% TFA Ion chromatography (acetate content): 7.1% acetate = 1.84 equivalents LC-MS (MCW-TOF-AQ-YMC-18 min): Rt=7.74 min; MS (ESIpos): m / z=725[M+2H] + Example 264 Sequence: ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH(L-tartrate) (Example 264) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidamido-propyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl)-5,8,11,14,17,25,28,35-octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2’,1’:13,14][1,2,5,8,11,14,17,20,23,26]dithiaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolinamide (2R,3R)-2,3-dihydroxybutanedioic acid (L-(+) tartaric acid) salt IRA410 chloride resin (12.5 g) was placed in an empty 20 mL Biotage column and the resin was washed with a 5% solution of ACN-water (10 times). The cartridge was attached to an automatic ion exchange station. The resin was washed with 20 column volumes of 1M NaOH solution (2×1800 s rotation speed 80 minutes -1 ) and then the column was washed with water until the pH of the eluent was <pH 9 (10 column volumes), and then the column was eluted with 1M L-tartaric acid solution (10 column volumes), and then the column was washed with water until the pH of the eluent was >5 (about 10 column volumes). The peptide ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH2 (TFA salt) (1200 mg) was loaded onto the column with 12 mL of 5% ACN / water, the column was circulated 10 times according to the general method F, and then the resin was washed 10 times with a 5% ACN / aqueous solution, and the eluate was collected in a 50 mL Falcon tube. The parameter settings used for this scale were as follows: Washing with H2O: Runtime 3085 s; 80 minutes -1 ; 1 cycle (is 90 ml volume) Sample circulation with peptide: Runtime 3085 s; 80 minutes -1 ; 1 cycle (is 180 ml volume) Washing with H2O (or % ACN in H2O: Runtime 3085 s; 80 minutes -1 ; 1 cycle (is 90 ml volume) The combined eluates were lyophilized to give 1170 mg (100% purity, 91% yield) of the target L-tartrate salt.

[0488] Ion chromatography (chloride content): <1% chloride Ion chromatography (TFA content): <1% TFA Ion chromatography (acetate content): 13.4% acetate = 1.49 equivalents LC-MS (MCW-TOF-AQ-YMC-18 min): Rt=7.62 min; MS (ESIpos): m / z=725[M+2H] + Example 376 Sequence: ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH(citrate) (Example 376) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl)-5,8,11,14, 17,25,28,35-Octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolinamide, 2-hydroxypropane-1,2,3-tricarboxylic acid (citric acid) salt [ka]

[0489] IRA410 chloride resin (26 g) was placed in an empty 20 mL Biotage column, and the resin was washed (10 times) with a 5% solution of ACN-water. The cartridge was then installed in an automated ion exchange station. The resin was washed with 20 column volumes of 1 M NaOH solution (2 x 1800 s, 80 min). -1) It was washed. The column was washed with water until the pH of the eluent became <pH 9 (10 column volumes). Then, the column was eluted with 1 M citric acid solution (10 column volumes), and then washed with water until the pH of the eluent became >5 (about 10 column volumes). Peptide (N-Me)GIC+SRSLP-(Oic)-I-Pen+IP-NH2 (TFA salt) (1200 mg) was loaded onto the column with 12 mL of 5% ACN / water, and the column was circulated 10 times according to General Method F. Then, the resin was washed 10 times with 5% ACN / aqueous solution, and the eluate was collected in a 50 mL Falcon tube. The parameter settings used for this scale were as follows: Washing with H2O: Runtime 3085 s; 80 minutes -1 ; 1 cycle (is 90 ml volume) Sample circulation using peptide: Runtime 3085 s; 80 minutes -1 ; 1 cycle (is 180 ml volume) Washing with H2O (or % ACN in H2O): Runtime 3085 s; 80 minutes -1 ; 1 cycle (is 90 ml volume) The combined eluate was lyophilized to obtain 1370 mg (purity 100%, yield 97%) of the target L-tartrate.

[0490] Ion chromatography (chloride content): <1% chloride Ion chromatography (TFA content): <1% TFA Ion chromatography (acetate content): 19.0% acetate = 1.77 equivalents LC-MS (MCW-TOF-AQ-YMC-18 minutes): Rt = 7.6 minutes; MS (ESIpos): m / z = 725 [M+2H] + Those skilled in the art recognize that the stoichiometry by ion chromatography is not always stoichiometric (e.g., 1:2 or 1:1), and the moisture content is not taken into account, which can be 8 - 15% moisture content if measured in some cases.

[0491] General Method G: Preparation of the Salt-Free Form The preparation of AIC+SRSLP-(Oic)-I-(Pen)+IPD-NH2 (salt-free form) (Example 27) is representative.

[0492] Five grams of the TFA salt (Example 75) was added to 70 ml of ethanol without the acid modifier. o The product was purified by reverse-phase HPLC using an acetonitrile-water gradient of C. The desired fractions were combined and lyophilized to give 2.2 grams of the salt-free form (Example 27); LC-MS (>99% purity); ion chromatography (<1% TFA). General method H: Large scale synthesis The synthesis of AIC+SRS-((tBu)A)-PPI-((N-Me)C)+IPD-NH2 (Example 48) is representative. [ka]

[0493] Peptides were synthesized using standard Fmoc chemistry.

[0494] 1) DMF was added to a container containing MBHA resin (180 mmol), and the resin was allowed to swell for 2 hours.

[0495] 2) Fmoc-Rink amide linker was added and mixed for 30 seconds, followed by saturation buffer. Mixing was continued for approximately 1 hour using N2 bubbles.

[0496] 3) The solution was drained from the resin, and the resin was washed with DMF (30 seconds x 3 times).

[0497] 4) A solution of 20% piperidine / DMF was added and mixed with nitrogen bubbling for 30 minutes.

[0498] 5) The solution was drained and the resin was washed with DMF (5 times).

[0499] 6) The Fmoc-amino acid solution was added and mixed for 30 seconds, followed by the activation buffer. Mixing was carried out with N2 bubbles for approximately 1 hour.

[0500] 7) Steps 3-6 were repeated for each amino acid coupling.

[0501] Table 10: Notes on materials and conditions used [Table 10]

[0502] A solution of 20% piperidine in DMF was used for 30 min for Fmoc deprotection. The coupling reaction was monitored by the ninhydrin test and the resin was washed five times with DMF.

[0503] Peptide cleavage and purification: 1) Cleavage buffer (92.5% TFA:2.5% EDT:2.5% TIS:2.5% HO) was added to the flask containing the side-chain protected peptide at room temperature, and the mixture was stirred for 2 hours.

[0504] 2) The peptide was precipitated with cold isopropyl ether and centrifuged (5000 rpm for 2 min).

[0505] 3) The peptide was washed two more times with isopropyl ether.

[0506] 4) The crude peptide was dried under vacuum for 2 hours.

[0507] Disulfide formation To a solution of the crude linear peptide (15 g, 9.92 mmol, 1.0 equiv) (20 batches) in HO / I (3 L 2:1) was added NHHCO (7.84 g, 99.2 mmol, 10 equiv). The mixture was stirred at 30 °C for 16 h, quenched by the addition of HCl to pH ∼6, and then lyophilized. The lyophilizate was purified by Prep-HPLC to give the desired Example 48 (41.8 g, 95.8%, 13.3% yield) as a white solid (156.8 g total).

[0508] Purification conditions Apparatus: Shimadzu 10A; peptide was dissolved in DMF / HO. Mobile phase: A (HO (0.075% TFA in HO), B EtOH; gradient: 10-50% elution, 60 min. Retention time: 47 min. Column: Luna, C18, 10 μm, 100 A, 25 cm × 50 mm; flow rate: 80 mL / min; wavelength: 220 / 254 nm; oven temperature: room temperature.

[0509] Method I: ((N-Me)A)-IC + SRSLP-(Oic)-I-(Pen) + IP-((N-benzyl)D)-NH 2 Adjustment of (Example 462) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methyl-L-alanyl-L-isoleucyl)amino]-6-(2- Methylpropyl)-5,8,11,14,17,25,28,35-octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolyl-N 4 -Benzyl-L-aspartamide [ka]

[0510] Peptides were prepared on Rink Amide MBHA resin (typically loading approximately 0.4-0.6 mmol / g) at a 0.3 mmol scale. The first amino acid, Fmoc-Asp(2-phenylisopropyl ester)-OH (CAS 200336-86-3), was added to the resin using the method described in Method C. The peptide sequence was assembled using the steps described in Method C. Once the sequence was complete, the 2-OPP protecting group was removed using 1% TFA in DCM for 30 minutes. N-benzylamine (3 equiv.), DIC (3 equiv.), and Oxyma (3 equiv.) were added, and the mixture was shaken for 2 hours. The peptide was cleaved, disulfide bonds formed, and purified according to Method C.

[0511] General Method J. Preparation of ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-(*Dap)-OH (Example 467) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl)-5,8,11 ,14,17,25,28,35-Octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-N-[(2S)-2-amino-2-carboxyethyl]-L-prolinamide [ka]

[0512] The peptide was prepared on 2-chlorotrityl resin according to Method C. BOC-Dap(Fmoc)-OH (CAS 122235-70-5) was loaded onto the resin according to Method C, and the peptide was assembled and purified according to Method C.

[0513] General Method K. Preparation of (2-(morpholine)acetyl)-IC + SRS-((tBu)A)-PPI-(Pen) + IPD-NH (Example 283) Example 27A N-(Bromoacetyl)-L-isoleucyl-L-cysteinyl-L-seryl-L-arginyl-L-seryl-4-methyl-L-leucyl-L-prolyl-L-prolyl-L-isoleucyl-3-sulfanyl-L-valyl-L-isoleucyl-L-prolyl-L-alpha-asparagine [ka]

[0514] The sequence IC+SRS-((tBu)A)-PPI-(Pen)+IPD-NH was prepared on Rinkamide ChemMatrix resin in 12 replicates at a 0.1 mmol scale according to Method A. To the resin-containing peptide (200 mg, 100 μmol) was added N,N'-diisopropylcarbodiimide (31 μL, 200 μmol), ethyl(hydroxyimino)cyanoacetate (28.4 mg, 200 μmol), and bromoacetic acid (14 μL, 200 μmol). The mixture was shaken overnight at room temperature in 5 mL of DMF on a thermomixer shaker. The resin was filtered, washed with DMF (6 × 5 mL), and then washed with DCM (6 × 5 mL), and then dried under vacuum for the next step.

[0515] Example 283 N-[(5aS,11S,14S,17S,20S,23R,28R,31S,33aS)-31-[(2S)-butan-2-yl]-17-(3-carbamimidamidopropyl)-11-(2,2-dimethylpropyl)-14,20-bis(hydroxymethyl)-27,27-dimethyl-23-({N-[(morpholin-4-yl)acetyl]-L-isopropyl] soleucyl}amino)-5,10,13,16,19,22,30,33-octaoxooctacosahydro-1H,5H,10H-dipyrrolo[2,1-j:2',1'-m][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosine-28-carbonyl]-L-isoleucyl-L-prolyl-L-alpha-asparagine [ka]

[0516] To Example 27a (200 mg, 74% purity, 77.7 μmol) in NMP (10 mL) was added morpholine (34 μl, 390 μmol), and the suspension was shaken in a thermomixer at 37° C. The solid resin was collected by filtration and washed twice with 10 mL of DMF and 10 mL of dichloromethane. The residue was treated with TFA / EDT / thionaisol (90:3:7) and shaken at room temperature for 2 hours. Cold diethyl ether (-10 o After treatment with HCl (C), the suspension was centrifuged under nitrogen and washed three times with cold diethyl ether (3 × 30 mL). 0.1 M aqueous ammonium bicarbonate solution (400 mL) was added to the residue, and the reaction mixture was shaken overnight in a round-bottom flask open to the air. The solution was lyophilized, and the lyophilized powder was purified according to Method A to give 20.4 mg of Example 283 (98% purity, 16% yield).

[0517] Example 300 (2-Hydroxyacetyl)-IC+SRS-((tBu)A)-PPI-(Pen)+IPD-NH2 was isolated as a by-product.

[0518] General Method L. Preparation of Sodium and Choline Salt Forms These salts were prepared from the unsalted form of the peptide. The following two examples are representative of the general method: Example 431 N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-18-[(L-alanyl-L-isoleucyl)amino]-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,11,14, 17,25,28,35-Octaoxodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-N-[(1S)-1,2-dicarboxylatoethyl]-L-prolinamide, sodium salt [ka]

[0519] Sequence: AIC+SRSLP-(Oic)-I-(Pen)+IPD-OH (sodium salt) (Example 431) To a solution of AIC + SRSLP-(Oic)-I-(Pen) + IPD-OH (free salt form) (100 mg) in 20 mL of acetonitrile / water (1:1), 1.3 mL of 0.1 M sodium hydroxide solution was added. The mixture was dried under vacuum to give 109 mg (104% yield, 99% purity) of the target compound.

[0520] LC-MS (MCW-TOF-AQ-YMC-18 min): t =8.62 min;MS(ESIpos):m / z=783[M+2H]2+ Ion chromatography (sodium content): 2.9% sodium = 2.03 equivalents Example 432 N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-18-[(L-alanyl-L-isoleucyl)amino]-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,11,14,17,25,28,35-octaoxa Xodotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-N-[(1S)-1,2-dicarboxylatoethyl]-L-prolinamide, 2-hydroxy-N,N,N-trimethylethane-1-ammonium)(choline) salt [ka]

[0521] To a solution of AIC + SRSLP-(Oic)-I-(Pen) + IPD-OH (free salt form) (100 mg) in 20 mL of acetonitrile / water (1:1), 511 μL of 0.25 M choline hydroxide solution was added. The mixture was dried under vacuum to give 122 mg (105% yield, 99% purity) of the target compound.

[0522] LC-MS (MCW-TOF-AQ-YMC-18 min): t =8.65 minutes;MS(ESIpos):m / z=783[M+2H]2+ Ion chromatography (choline content): 11.7% choline = 1.97 equivalents Example 13 AIC + SRSLP-(Oic)-I-(Pen) + IPD-OH (Example 13) (3S)-4-amino-3-({(2S)-1-[(2S,3S)-2-{[(2R)-2-{[(2S,3S)-2-{[(2S,3aS,7aS)-1-{(2S)-1-[(2S,5S,8S,11S,14R,17S,20S)-20-amino-17-[(2S)-butan-2-yl]-8-(3-carbamimidamidopropyl)-5,11-bis(hydroxymethyl)-2-(2-methylpropyl)-4,7,1 0,13,16,19-Hexaoxo-14-(sulfanylmethyl)-3,6,9,12,15,18-hexaazahenicosanan-1-oyl]pyrrolidine-2-carbonyl}octahydro-1H-indole-2-carbonyl]amino}-3-methylpentanoyl]amino}-3-methyl-3-sulfanylbutanoyl]amino}-3-methylpentanoyl]pyrrolidine-2-carbonyl}amino)-4-oxobutanoic acid [ka]

[0523] Peptide synthesis Peptides were synthesized using standard Fmoc chemistry.

[0524] 1) Preparation of resin: To CTC resin (15.0 mmol, 15.0 g, 1.0 mmol / g) was added Fmoc-Asp(OtBu)-OH (15.0 mmol, 6.17 g, 1.0 equiv.) and DIEA (10.4 mL, 60.0 mmol, 4.0 equiv.) in DCM (300 mL). The mixture was stirred with N2 for 2 hours at 20 °C. Methanol (15.0 mL) was then added, and the resin was stirred with N2 for another 30 minutes. The resin was washed with DMF (300 mL × 3). A 20% piperidine solution in DMF (300 mL) was then added, and the mixture was stirred with N2 for 20 minutes at 20 °C. The mixture was then filtered to obtain the resin. The resin was washed with DMF (300 mL × 3) and filtered to obtain the resin.

[0525] 2) Coupling: Fmoc-Pro-OH (15.2 g, 45.0 mmol, 3.0 equiv), HBTU (16.2 g, 42.8 mmol, 2.85 equiv), and DIEA (90.0 mmol, 15.6 mL, 6.00 equiv) in DMF (100 mL) were added to the resin, and the resin was agitated with N for 30 min at 20 °C. Then, the resin was washed with DMF (300 mL × 5).

[0526] 3) Deprotection: 20% piperidine in DMF (300 ml) was added to the resin and the mixture was stirred with N2 at 20° C. for 20 minutes.

[0527] 4) Steps 2 and 3 were repeated for all other amino acids: Table 11: Reagents used in Example 13 [Table 11]

[0528] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin (all amino acids except Pro) and chloranil test (Pro), and the resin was washed five times with DMF (300 mL).

[0529] HBTU, DIEA Supplier: Suzhou Highfine Biotech Co, Ltd. Peptide cleavage and purification 5) The resin was washed with MeOH (300 mL x 5) and dried under vacuum to give 41.5 g of peptide resin. Then, 400 ml of cleavage buffer (92.5% TFA / 2.5% 3-mercaptopropionic acid / 2.5% TIS / 2.5% HO) was added to the flask containing the side-chain protected peptide resin at 20°C, and the mixture was stirred for 2.5 hours and then filtered.

[0530] 6) The peptide was precipitated with cold tert-butyl methyl ether (4000 mL) and centrifuged (3000 rpm for 3 minutes) to obtain crude solid peptide. The crude peptide precipitate was washed three more times with tert-butyl methyl ether (1000 mL × 3). The crude peptide was then dried under vacuum for 2.0 hours to obtain 21.8 g of crude peptide.

[0531] 7) The crude peptide (21.8 g) was dissolved in CHCN (6.5 L) and water (6.5 L), and I (0.1 M in MeOH, 55 mL) was added at 20° C. until the yellow color persisted. The mixture was then stirred at 20° C. for 2 minutes. Sodium thiosulfate solution (0.1 M in water, 15 mL) was then added dropwise until the yellow color disappeared. The mixture was then lyophilized to give a crude powder (22.7 g).

[0532] 8) Purification of the crude peptide by preparative HPLC (conditions: A: 0.075% TFA in water B: CH3CN), collection of product-containing fractions, and lyophilization gave the target peptide (sequence ID13) as a white solid. (4305.1 mg, 18.3% yield, 99.1% by 110 Å C18 RP-HPLC column (Gemini C18 5 μm 110 Å 150x4.6 mm); 97.8% by 300 Å C18 RP-HPLC column (Discovery BIO Wide Pore C18 5 μm 300 Å 150x4.6 mm), TFA salt).

[0533] Table 12: Purification conditions [Table 12]

[0534] LC-MS:Rt=1.369min;1564[M+2H]+ Rt=8.72 min (purity 99.1%) Gemini C18 5μm 110Å 150x4.6mm Rt=9.42min(97.8%) Discovery BIO Wide Porei C18 5μm 300Å 150x4.6mm From Example 13 (TFA salt), Example 29 (HCl salt) can be prepared using general method F; Example 5 (unsalted form) can be prepared using general method G. The preparation of Example 431 (sodium salt) and Example 432 (choline salt) from Example 5 are shown below as Example 431 and Example 432, respectively.

[0535] Example 237 AIC + SRSL-(L-dehydroproline)-(Oic)-I-(Pen) + IPD-OH (Example 237) N-[(6S,9S,12S,15S,18R,23R,26S,28aS,35aS)-18-[(L-alanyl-L-isoleucyl)amino]-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,11,14,17,25,28,35-octaoxo-6,7,8,9,10,11,12,13 ,14,15,16,17,18,19,23,24,25,26,27,28,28a,29,29a,30,31,32,33,33a,35,35a-Triacontahydro-3H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolyl-L-aspartic acid [ka]

[0536] The peptide was prepared according to Method C (3.12 g). Analytical data for this peptide are shown in Tables 16 and 17.

[0537] Example 142 AIC+SRSL-(Pro-D2)-(Oic)-I-(Pen)+IPD-OH(Example 142) N-[(1R,2S,6S,9S,12S,15S,18R,23R,26S,28aS,35aS)-18-[(L-alanyl-L-isoleucyl)amino]-26-[(2S)-butan-2-yl]-12-(3-carbamimidamidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,11,14,17,25,28,35-octaoxo(1,2- 2 H2) dotriacontahydro-1H,5H,22H-pyrrolo[2',1':13,14][1,2,5,8,11,14,17,20,23,26]dithiaoctaazacyclononacosino[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolyl-L-aspartic acid [ka]

[0538] The peptide from Example 237 (30 mg, 19.2 μmol) was dissolved in methanol-d4 (3 mL) (99.8% purity), and rhodium black (CAS 7440-16-6, ACBR, Article No. AB25746) (10 mol%, 1.9 μmol) was added to the solution in an M-Braun glovebox under argon. The 8 ml reaction vial was placed in an MTP pressure reactor block placed on a Chemspeed Swing XL platform and enclosed in an M-Braun glovebox (Chemspeed Technologies AG, Woelferstrasse 8, 4414 Fuellinsdorf, Switzerland, Chemspeed@Chemspeed.com). The reaction was shaken at an orbital speed of 650 rpm at room temperature under 3 bar deuterium gas pressure for 16 hours overnight. The pressure block was purged with argon, and the vial was removed from the reactor block. Methanol-d4 was evaporated on a rotary evaporator, and the resulting solid was dissolved in water (2 mL) and stirred for 1 h to exchange exchangeable deuterium atoms for hydrogen. After 1 h, acetonitrile was added, and the product was purified by reverse-phase HPLC [Column: Phenomenex Aeris Peptide 5μ XB-C18, AXIA Packed, 21.2 x 250 mm + 5μ cartridge, Flow rate: 20 mL / min, Focused gradient using HO / ACN (with 0.1% TFA as an acidity modifier). FOK30-40: 0 min 5% ACN, 0-13 min ramp to 30% ACN, 13-39 min flat gradient to 40% ACN, 39-46 min 40% ACN homogeneous]. The chromatographic fractions containing the product were analyzed by analytical HPLC (converging gradient) and LC-MS and pooled accordingly to give two fractions of pure product: 11.4 mg (90% purity, 34% yield) and 4.2 mg (99% purity, 14% yield). The analytical data for this peptide are shown in Table 16.

[0539] The resulting peptide was digested with trypsin, followed by reaction with dithiothreitol, and the resulting fragments were analyzed by TOF-MS-MS. Trypsin digestion cleaved the peptide between arginine and serine (accurate mass 1583.8384, found 1583.852 [M+2H]2+). Reaction with dithiothreitol produced two fragments, AICSR-OH (exact mass 548.2741, found 548.283 [M+H]) and SL-(Pro-D2)-(Oic)-I-(Pen)+IPD-OH (exact mass 1037.5800, found 1038.587 [M+H]), confirming the deuterium position in the 838.47 amu ((Pro-D2)-(Oic)-I-(Pen)+IPD-OH) and 738.40 amu SL-((Oic)-I-(Pen)+IPD-OH) fragments.

[0540] Example 28A (2S,3aS,6aS)-Octahydrocyclopenta[b]pyrrole-2-carboxylic acid-hydrogen chloride (1 / 1) (single stereoisomer) [ka]

[0541] The reaction was carried out under an argon atmosphere. To palladium on carbon (200 mg, 10%) was added 20 mL of methanol and benzyl (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylate-hydrogen chloride (1 / 1) (1.00 g, 3.55 mmol). The reaction flask was charged with hydrogen gas (1 atm), and the reaction mixture was stirred under a hydrogen atmosphere at room temperature overnight. The reaction mixture was filtered through diatomaceous earth, and the filter cake was washed with methanol. The filtrate was evaporated and dried under vacuum to give 698 mg of the target compound.

[0542] (2S,3aS,6aS)-Octahydrocyclopenta[b]pyrrole-2-carboxylic acid is also commercially available in the salt-free form.

[0543] Example 29A (2S,3aS,6aS)-1-{[(9H-fluoren-9-yl)methoxy]carbonyl}octahydrocyclopenta[b]pyrrole-2-carboxylic acid (single stereoisomer) [ka]

[0544] To (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid-hydrogen chloride (1 / 1) (680 mg, 3.55 mmol) in 12 mL of water was added a solution of 1-({[(9H-fluoren-9-yl)methoxy]carbonyl}oxy)pyrrolidine-2,5-dione (Fmoc-OSu) (1.26 g, 3.73 mmol) in 18 mL of acetone with sodium bicarbonate (2.98 g, 35.5 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was treated with water, acidified with 1 M hydrochloric acid, and extracted twice with ethyl acetate. The organic phase was washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, and evaporated. The residue was separated by silica gel chromatography (Biotage Isolera, column: 50 g SNAP Ultra; eluent: dichloromethane / methanol 100:2 to 100:5). The appropriate fractions were combined, evaporated and dried under vacuum to give 1.27 g (99% purity, 94% yield) of the target compound.

[0545] LC-MS (Method 10): R t = 2.08 min; MS (ESI pos): m / z = 378 [M+H] + 1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 1.260 (1.76), 1.274 (1.85), 1.299 (1.78), 1.321 (2.09), 1.335 (2.38), 1.440 (0.92), 1.455 (1.10), 1.473 (0.87), 1.521 (3.61), 1.537 (3.86), 1.551 (2.28), 1.607 (4.53), 1.620 (3.68), 1.640 (3.08), 1.653 (1.66), 1.712 (2.02), 1.781 (1.38), 1.816 (2.30), 1.827 (2.26), 1.839 (2.37), 1.855 (1.68), 2.073 (1.20), 2.305 (0.83), 2.328 (1.99), 2.351 (1.35), 2.360 (1.64), 2.384 (0.92), 2.468 (0.60), 2.606 (1.50), 2.670 (1.11), 3.860 (0.94), 3.874 (1.94), 3.893 (1.95), 3.907 (0.92), 4.143 (2.61), 4.156 (3.68), 4.167 (5.16), 4.186 (5.89), 4.266 (1.43), 4.280 (3.20), 4.294 (2.11), 4.345 (1.31), 4.371 (3.19), 4.386 (3.83), 4.396 (4.39), 4.411 (3.74), 4.422 (2.40), 4.437 (0.98), 7.292 (0.97), 7.311 (3.89), 7.329 (6.99), 7.346 (6.08), 7.363 (2.49), 7.395 (9.91), 7.414 (16.00), 7.432 (6.98), 7.663 (9.73), 7.680 (6.46), 7.879 (12.31), 7.897 (11.49), 12.586 (0.80). Example 30A (1R,3S,5R)-2-Azabicyclo[3.1.0]hexane-3-carboxylic acid (single stereoisomer) [ka]

[0546] (1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (CAS RN: 197142-34-0, 500 mg) was dissolved in 4 M HCl-dioxane solution (3 mL) and stirred at room temperature for 8 h. The solvent was evaporated and the crude product was used directly in the next step.

[0547] Example 31A (1R,3S,5R)-2-{[(9H-fluoren-9-yl)methoxy]carbonyl}-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (single stereoisomer) [ka]

[0548] To a solution of (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (Example 30A, 280 mg, 2.20 mmol) in 3 mL of water was added sodium bicarbonate (1.85 g, 22.0 mmol) and a solution of 1-({[(9H-fluoren-9-yl)methoxy]carbonyl}oxy)pyrrolidine-2,5-dione (0.78 g, 2.31 mmol) in 4.5 mL of acetone. The reaction mixture was stirred overnight at room temperature. The reaction mixture was treated with water and extracted twice with ethyl acetate. The organic phase was washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by preparative reverse-phase HPLC (column: Reprosil; C18; 10 μm; 125 × 30 mm; eluent A: CAN, eluent B: water containing 0.1% formic acid), and the product-containing fractions were combined and lyophilized to give 88.0 mg (96% purity, 11% yield) of the target compound.

[0549] LC-MS (Method 11): R t = 1.32 min; MS (ESI pos): m / z = 372 [M+Na]+ 1 H-NMR (400 MHz, DMSO-d6) δ [ppm]: -0.149 (1.15), -0.008 (10.73), 0.008 (9.13), 0.146 (1.21), 0.468 (1.54), 0.534 (3.66), 0.803 (4.38), 1.682 (2.68), 2.072 (0.75), 2.128 (2.19), 2.142 (2.32), 2.223 (1.05), 2.293 (2.19), 2.327 (4.45), 2.349 (2.68), 2.366 (2.75), 2.523 (3.89), 2.670 (1.44), 2.710 (1.31), 2.816 (0.82), 3.413 (3.93), 3.757 (1.83), 4.010 (2.91), 4.140 (2.39), 4.211 (3.11), 4.285 (12.30), 6.929 (0.43), 6.950 (0.43), 7.340 (9.91), 7.358 (6.02), 7.406 (10.08), 7.425 (16.00), 7.443 (7.10), 7.650 (2.85), 7.702 (4.52), 7.724 (5.53), 7.746 (3.40), 7.892 (12.40), 7.910 (9.85), 12.680 (0.65). This amino acid was used for Fmoc SPPS and is also commercially available.

[0550] Example 32A Benzyl (2R,4R)-4-[(2S)-butan-2-yl]-2-tert-butyl-5-oxo-1,3-oxazolidine-3-carboxylate (single stereoisomer) [ka]

[0551] To a suspension of D-alloisoleucine (8.06 g, 61.4 mmol) in 120 mL of ethanol was added a sodium hydroxide solution prepared from 2.45 g of sodium hydroxide in 11 mL of water. The reaction mixture was stirred at room temperature for 90 minutes and then evaporated. The residue was treated with 195 mL of pentane, followed by the addition of 2,2-dimethylpropanal (10 mL, 92 mmol). The reaction mixture was stirred at 45 °C for 24 hours using a Dean-Stark trap attached to a round-bottom flask. After cooling, the solvent was evaporated, and the residue was dried several times with toluene (azeotropic distillation). The residue was treated with 165 mL of dichloromethane and cooled to 0 °C. Benzyl carbonochloridate (13 mL, 92 mmol) was added, and the reaction mixture was stirred at 0-4 °C for 1 week. 4-Dimethylaminopyridine (375 mg, 3.1 mmol) and 65 mL of water were added to the reaction mixture, and the reaction mixture was allowed to warm to room temperature and then stirred overnight at room temperature. The reaction mixture was treated several times with ethyl acetate and extracted. The combined organic phases were washed twice with 10% aqueous citric acid and then twice with saturated aqueous sodium bicarbonate. The aqueous phase was re-extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, and evaporated. The residue was purified by silica gel chromatography (Isolera; eluent: cyclohexane / ethyl acetate 100:3 to 100:7). The product-containing fractions were combined, evaporated, and then dried under vacuum to give 8.71 g of the target compound.

[0552] Example 33A Benzyl (2R,4R)-4-[(2S)-butan-2-yl]-2-tert-butyl-4-methyl-5-oxo-1,3-oxazolidine-3-carboxylate (single stereoisomer) [ka]

[0553] The reaction was carried out under an argon atmosphere. Benzyl (2R,4R)-4-[(2S)-butan-2-yl]-2-tert-butyl-5-oxo-1,3-oxazolidine-3-carboxylate (Example 32A, 8.12 g, 24.4 mmol) in 100 mL of dry THF was cooled to -78 °C. Potassium hexamethyldisilazide (44 mL, 29 mmol) was added dropwise, and the reaction mixture was stirred at -78 °C for 2 hours. Iodomethane (2.3 mL, 37 mmol) was added dropwise, and the reaction mixture was stirred at room temperature for 2.5 hours. Saturated aqueous ammonium chloride solution (20 mL) was added to the reaction mixture, and after the reaction mixture was allowed to reach room temperature, the product was extracted into ethyl acetate. The combined organic phase was washed with saturated aqueous sodium bicarbonate, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (Isolera, column: 100 g SNAP Ultra; eluent: cyclohexane / ethyl acetate 100:3 to 100:7). The product-containing fractions were combined, evaporated, and then dried under vacuum to give 3.2 g of the target compound.

[0554] Example 34A N-[(benzyloxy)carbonyl]-2-methyl-D-alloisoleucine (single stereoisomer) [ka]

[0555] To a solution of benzyl (2R,4R)-4-[(2S)-butan-2-yl]-2-tert-butyl-4-methyl-5-oxo-1,3-oxazolidine-3-carboxylate (Example 33A, 3.20 g, 9.21 mmol) in methanol (40 mL) was added 1 M sodium hydroxide solution (40 mL). The reaction mixture was stirred under reflux for 2 hours. After cooling, the methanol was evaporated. The residue was treated with 10% citric acid solution, and the product was extracted into ethyl acetate. The aqueous phase was extracted twice more with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, evaporated, and then dried under vacuum to give 2.77 g of the target compound.

[0556] Example 35A 2-Methyl-D-alloisoleucine (single stereoisomer) [ka]

[0557] The reaction was carried out under an argon atmosphere. Palladium on carbon (390 mg, 10%) was added to N-[(benzyloxy)carbonyl]-2-methyl-D-alloisoleucine (Example 34A, 2.57 g, 9.20 mmol) in 190 mL of methanol. The reaction mixture was stirred under a hydrogen atmosphere at room temperature and atmospheric pressure for 4 hours. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated and then dried under vacuum to give 1.32 g (98% yield) of the target compound.

[0558] Example 36A N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-2-methyl-D-alloisoleucine (single stereoisomer) [ka]

[0559] To a solution of 2-methyl-D-alloisoleucine (Example 35A, 1.32 g, 9.09 mmol) in a mixture of water (20 mL) and acetone (29 mL) was added sodium bicarbonate (7.64 g, 90.9 mmol), followed by a solution of 1-({[(9H-fluoren-9-yl)methoxy]carbonyl}oxy)pyrrolidine-2,5-dione (3.22 g, 9.55 mmol) in acetone (20 mL). The reaction mixture was stirred overnight at room temperature. The reaction mixture was treated with MTBE and extracted. The aqueous phase was acidified (pH 1) with concentrated hydrochloric acid and extracted three more times with ethyl acetate. The combined organic phases were dried over magnesium sulfate, filtered, and evaporated. The residue was separated by silica gel chromatography (Isolera, column: 50 g SNAP Ultra; eluent: dichloromethane / methanol 100:3 to 100:5). The appropriate fractions were combined, evaporated and then dried under vacuum.

[0560] The MTBE phase was then treated with 5% sodium bicarbonate solution and extracted with ethyl acetate (600 mL). The aqueous phase was acidified with concentrated hydrochloric acid (pH 1) and extracted with ethyl acetate, respectively. The combined organic phases were dried over magnesium sulfate, filtered, evaporated, and then dried under vacuum. The combined crude product yield was 2.53 g.

[0561] The residue was purified by silica gel chromatography (Isolera, column: 50 g SNAP Ultra; eluent: dichloromethane / methanol 97:3 to 97:5). The product-containing fractions were combined, evaporated, and then dried under vacuum to give 967.4 mg of the target compound. The product was further dissolved in water / acetonitrile and lyophilized before use.

[0562] LC-MS (Method 10): R t = 2.10 min; MS (ESI pos): m / z = 368.1857 [M+H] + This amino acid was used for Fmoc SPPS.

[0563] Example 37A (6S)-1,1-Difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (mixture of stereoisomers) [ka]

[0564] To (6S)-5-(tert-butoxycarbonyl)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (Omega, CAS 1357482-03-1) (2.50 g, 9.02 mmol) in 85 mL of dichloromethane was added trifluoroacetic acid (15 mL, 250 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and dried under vacuum for the next step.

[0565] Example 38A (6S)-5-{[(9H-Fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (mixture of stereoisomers) [ka]

[0566] To a solution of (6S)-1,1-difluoro-5-azapyrro[2.4]heptane-6-carboxylic acid (mixture of stereoisomers) (Example 37A, 1.60 g, 9.03 mmol) in water (13 mL), sodium bicarbonate (7.59 g, 90.3 mmol) was added, followed by a solution of 1-({[(9H-fluoren-9-yl)methoxy]carbonyl}oxy)pyrrolidine-2,5-dione (3.20 g, 9.48 mmol) in acetone (20 mL). The reaction mixture was stirred overnight at room temperature. The reaction mixture was treated with water and extracted twice with MTBE. The aqueous phase was acidified to pH 3 with hydrochloric acid and extracted three times with dichloromethane. The organic phase was washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, evaporated, and dried under vacuum to give 3.31 g (92% yield) of the target compound.

[0567] LC-MS (Method 10): R t = 1.99 points; MS (ESI pos): m / z = 400 [M+H] + 1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 1.106 (16.00), 1.589 (0.92), 1.605 (1.39), 1.619 (1.06), 1.627 (1.11), 1.655 (0.62), 1.682 (1.25), 1.703 (1.21), 1.916 (0.49), 1.949 (0.86), 1.978 (0.91), 2.012 (0.85), 2.042 (0.52), 2.086 (11.35), 2.558 (0.83), 2.568 (0.67), 2.591 (0.67), 2.645 (0.52), 2.670 (0.56), 3.077 (5.29), 3.442 (0.43), 3.452 (0.50), 3.471 (1.17), 3.479 (1.13), 3.500 (1.39), 3.527 (0.51), 3.562 (1.21), 3.627 (1.20), 3.652 (1.65), 3.677 (0.78), 4.186 (1.02), 4.200 (2.72), 4.223 (2.09), 4.238 (2.18), 4.257 (1.63), 4.279 (2.63), 4.294 (2.46), 4.306 (1.36), 4.322 (1.56), 4.348 (2.13), 4.359 (1.17), 4.372 (1.33), 4.382 (0.84), 4.477 (0.57), 4.499 (0.61), 4.510 (0.69), 4.518 (0.72), 4.533 (0.73), 4.541 (0.64), 5.754 (0.94), 7.300 (0.77), 7.316 (3.26), 7.334 (5.65), 7.353 (3.32), 7.406 (4.71), 7.425 (7.78), 7.443 (3.45), 7.640 (4.69), 7.659 (4.35), 7.683 (0.73), 7.884 (5.08), 7.903 (4.79). Example 39A (6S)-5-{[(9H-Fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1) and (enantiomer 2) [ka]

[0568] The diastereomeric mixture was separated using a THAR Supercritical Fluid Chromatography Prep 200 instrument using a Chiralpcel AD-H (SFC) 5 μm, 250 × 30 mm column; eluent: CO₂ / isopropanol (83:17); pressure: 135 bar; eluent temperature: 38 °C; cyclone temperature: 40 °C; cyclone pressure: 24 bar; flow rate: 100 g / min; detection: UV 210 nm; injection volume: 0.4 mL / injection; under normal operating conditions, 1 g / min of CO₂ is equivalent to approximately 1 mL / min. Sequence setting: Cycle time = 11.2 minutes Fraction 1 was collected between 5.85 and 8.0 minutes (enantiomer 1). Fraction 2 was collected between 8.4 and 10.25 minutes (enantiomer 2). The fractions were concentrated.

[0569] Fractions were analyzed using analytical SFC-MS (Agilent, column: Chiralpak AD-3 3 μm 100 × 4,6 mm, eluent: CO2 / iso-propanol (85:15); BPR pressure: 130 bar; BPR temperature: 60 °C; column temperature: 40 °C; flow rate: 3 mL / min; UV 210 nm).

[0570] Fraction 1 (rt = 2.081 min) was assigned the addition of "(Enantiomer 1)".

[0571] Fraction 2 (rt = 2.838 min) was assigned to the addition of "(enantiomer 2)".

[0572] The stereochemistry of the two enantiomers was not assigned.

[0573] Example 40A (3R*,6S)-5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azapyrro[2.4]heptane-6-carboxylic acid (single enantiomer, stereochemistry at enantiomer 1 and 3 is undefined) [ka]

[0574] Chiral SFC preparative purification gave 2,015 g of the title compound (100% ee).

[0575] LC-MS (Method 10): R t = 1.99 min; MS (ESI pos): m / z = 400 [M+H] + 1H-NMR (400 MHz, DMSO-d6) δ [ppm]: -0.008 (0.70), 0.008 (0.74), 1.032 (15.85), 1.047 (16.00), 1.285 (1.59), 1.568 (0.69), 1.589 (1.52), 1.617 (1.57), 1.635 (0.96), 1.656 (0.63), 1.686 (1.36), 1.702 (1.37), 1.941 (0.90), 1.974 (1.05), 2.008 (0.87), 2.041 (0.90), 2.073 (1.11), 2.557 (1.08), 2.566 (0.95), 2.591 (0.80), 2.645 (0.71), 2.670 (0.95), 2.701 (0.65), 3.451 (0.87), 3.468 (1.64), 3.479 (1.88), 3.495 (1.12), 3.505 (1.17), 3.626 (2.11), 3.651 (2.91), 3.677 (1.38), 3.770 (0.41), 3.778 (0.42), 4.169 (0.43), 4.186 (1.17), 4.200 (3.11), 4.221 (2.05), 4.238 (1.40), 4.256 (1.57), 4.279 (3.79), 4.293 (3.74), 4.304 (1.62), 4.322 (2.70), 4.336 (1.12), 4.348 (2.93), 4.358 (1.92), 4.371 (1.64), 4.381 (1.52), 4.509 (1.14), 4.517 (1.25), 4.531 (1.28), 4.540 (1.12), 7.300 (0.91), 7.315 (3.87), 7.334 (7.04), 7.350 (3.82), 7.352 (4.21), 7.406 (5.88), 7.424 (9.76), 7.443 (4.35), 7.640 (7.54), 7.658 (6.74), 7.885 (6.80), 7.904 (6.34), 10.194 (0.55), 10.991 (0.42). This amino acid was used for Fmoc SPPS.

[0576] Example 41A (3S*,6S)-5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azapyrro[2.4]heptane-6-carboxylic acid (single enantiomer, enantiomer 2, stereochemistry at the cyclopropyl center not assigned) [ka]

[0577] Chiral SFC preparative purification gave 1.041 g of the title compound (94.58% ee).

[0578] LC-MS (Method 10): R t = 1.93 min; MS (ESI pos): m / z = 400 [M+H] + 1H-NMR (400 MHz, DMSO-d6) δ [ppm]: -0.008 (1.19), 0.008 (...

Claims

1. A compound comprising a peptide or a pharma- ceutically acceptable salt, solvate or solvate of the salt thereof, The peptide is 【Chemistry 1】 A compound represented by the formula:

2. 2. A compound comprising the peptide of claim 1 or a pharma- ceutically acceptable salt, solvate or solvate of said salt, which acts as a MASP-1 and / or MASP-2 inhibitor and / or inhibits C3 deposition.

3. 3. A method for preparing a compound containing the peptide of claim 1 or 2, or a pharma- ceutically acceptable salt, solvate or solvate of a salt thereof, said method using solid phase peptide synthesis.

4. 3. A compound comprising a peptide according to claim 1 or 2, or a pharma- ceutically acceptable salt, solvate or solvate of the salt thereof, for use in the prevention and / or treatment of a disease.

5. 3. A compound comprising the peptide according to claim 1 or 2 or a pharma- ceutically acceptable salt, solvate or solvate of said salt for use in the prophylaxis and / or treatment of cardiovascular and cardiopulmonary disorders, crush injury, inflammatory disorders, cardiovascular, pulmonary, cerebral and renal sepsis sequelae, ischemia and / or reperfusion related injury, acute kidney injury, graft protection and delayed graft function, diseases of the blood and blood forming organs and immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, diseases of the respiratory system, digestive system or genitourinary system, sequelae of burns and trauma.

6. A pharmaceutical composition comprising at least one compound containing the peptide according to claim 1 or 2, or a pharma- ceutically acceptable salt, solvate or solvate of the salt thereof, in combination with one or more inert, non-toxic, pharma- ceutically suitable excipients.

7. 3. A pharmaceutical composition comprising at least one compound containing a peptide according to claim 1 or 2 or a pharma- ceutically acceptable salt, solvate or solvate of said salt in combination with one or more further active ingredients selected from the group consisting of inhibitors of phosphodiesterases, stimulators or activators of guanylate cyclase, IP receptor agonists, mineralocorticoid receptor antagonists, diuretics, PPAR-gamma agonists, PPAR-delta agonists, corticosteroids, active ingredients which reduce damage to organs under oxidative stress, compounds which inhibit the induction of cell death and apoptotic pathways, compounds which inhibit inflammatory responses and T-cell proliferation, antithrombotic agents, platelet aggregation inhibitors, thrombin inhibitors, GPIIb / IIIa antagonists, factor Xa inhibitors, heparin or low molecular weight (LMW) heparin derivatives and inhibitors of coagulation factor XI.

8. 8. A pharmaceutical composition according to claim 6 or 7 for the prevention and / or treatment of cardiovascular and cardiopulmonary disorders, crush injury, inflammatory disorders, sequelae of cardiovascular, pulmonary, cerebral and renal sepsis, ischemia and / or reperfusion related damage, acute kidney injury, graft protection and delayed graft function, diseases of the blood and blood forming organs and immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, diseases of the respiratory system, digestive system or genitourinary system, sequelae of burns and trauma.

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