MASP inhibitory compounds and uses thereof

Abstract

The present invention relates to novel mannose-binding lectin (MBL)-associated serine protease (MASP) inhibitory compounds and their analogs and derivatives, methods for preparing the same, their use alone or in combination for treating and / or preventing diseases, and their use in preparing drugs for treating and / or preventing diseases, especially for treating and / or preventing kidney and cardiovascular diseases and ischemia-reperfusion injury.

Description

[0001] The present invention relates to novel mannose-binding lectin (MBL)-related serine protease (MASP) inhibitory compounds and their analogs and derivatives, methods for their preparation, their use alone or in combination for treating and / or preventing diseases, and their use in preparing medicaments for treating and / or preventing diseases, in particular for treating and / or preventing 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. The complement system is an important component of innate immunity and is crucial for defending against invading pathogens and clearing dead cells and virus-infected cells. It builds a bridge between the innate immune response and the adaptive immune response. Activation of the complement system is also involved in the pathology of sepsis and ischemia-reperfusion injury (for example, after myocardial infarction, ischemic renal 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 and, under normal conditions, has a sentinel function against invading pathogens and dead cells by recognizing foreign and altered carbohydrate surface patterns and modifying their surfaces, respectively. Mannose-binding lectin (MBL), ficolin, and collectin are the main representatives of these lectins produced in the liver, kidney, and other organs (Garred et al., A journey through the lectin pathway of complement-MBL and beyond. Immunol Rev. 2016; 274(1): 74-97). After their deposition, they further recruit (recruitment) the zymogens of essentially two closely related serine proteases from the bloodstream, forming a complex with the mannose-binding lectin-associated serine proteases 1 and 2 (MASP-1 and MASP-2), in which the zymogens are close to each other. The current theory is that under in vivo conditions, the MASP-1 zymogen activates itself after recruitment and then activates the MASP-2 zymogen by cleavage. The activated MASP-1 further cleaves complement factor C2 into C2a and C2b. Activated MASP-2 also cleaves C2 and complement factor C4 into C4a and C4b, which, together with C2a, form the C4bC2a complex, which acts as the complement factor C3 convertase. The combination of C3 convertase activity and the continuous deposition of target cell C3 on the target cell surface represents a point of convergence for all three complement pathways, activating a common downstream cascade leading to the production of inflammatory mediators and target cell lysis.In intact human serum, the activity of both MASP enzymes is essential for the formation of C3 convertase (Héja et al., Revised mechanism of complement lectin-pathway activation revealing the role of serine protease MASP-1 as the exclusive activator of MASP-2. Proc Natl Acad Sci US A. 2012; 109(26): 10498-503).

[0003] The microvasculature plays a crucial role in inflammatory and ischemic organ pathologies. Barrier function, leukocyte trafficking, and coagulation control are critically dependent on the integrity of the luminal endothelial cell surface in small vessels. The luminal endothelial surface is lined with a dense, glycosylated layer derived from membrane-integrated glycoproteins, proteoglycans, and glycolipids, collectively known as the glycocalyx. Electron microscopic analysis of animal and human pathological specimens has demonstrated that the endothelial glycocalyx, in particular, undergoes rapid and radical degradation in response to ischemic challenges and inflammatory conditions such as sepsis. These changes expose carbohydrate remnants to the bloodstream that are undetectable under normal conditions (for review, see Sieve et al., Regulation and function of endothelial glycocalyx layer in vascular diseases. Vascul Pharmacol. 2018;100:26-33). Among other changes on the cell surface, altered carbohydrate patterns are thought to activate the lectin pathway, leading to the deposition of pattern-recognition lectins, subsequent C3 deposition, and the initiation of cell lysis. MBL and C3 deposition has been shown to occur after ischemia and acute kidney injury in species including humans. Activation of the lectin pathway is particularly relevant to reperfusion injury, as targeted deletion of MBL and MASP-2 protects mice from ischemia-reperfusion injury in the kidneys, heart, and intestine (Moller-Kristensen et al., Mannan-binding lectin recognizes structures on ischaemicreperfused mouse kidneys and is implicated in tissue injury. Scand J Immunol. 2005; 61(5): 426-34; Schwaeble et al., Targeting of mannan-binding lectin-associated serine protease-2 confers protection from myocardial and gastrointestinal ischemia / reperfusion injury. Proc Natl Acad Sci US A. 2011; 108(18): 7523-8).Furthermore, the absence of collectin-11, another MASP-activating lectin expressed primarily in the kidney, renders mice resistant to ischemic acute kidney injury (Farrar et al., Collectin-11 detects stress-induced L-fucose pattern to trigger 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 starting points. 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 with phage display selected peptides against mannose-binding lectin-associated serine protease (MASP)-1 and -2: significant contribution of MASP-1 to lectin pathway activation. J Immunol. 2010; 185(7): 4169-78; Héja et al., Monospecific inhibitors show that both mannose-binding lectin-associated serine protease-1 (MASP-1) and are essential for lectin pathway activation and reveal structural plasticity of MASP-2. J Biol Chem. 2012; 287(24): 20290-300). However, there is no evidence for the pharmaceutical utility and in vivo efficacy of these peptide inhibitors. Similarly, antibodies against MASP-2 that interfere with the interaction with the MASP zymogen have been identified and are in clinical development for atypical hemolytic uremic syndrome and other inflammatory kidney diseases (ClinicalTrials.gov Identifiers: NCT03205995; NCT02682407; NCT03608033). However, clinical evidence of their utility for preventing or treating acute, particularly ischemic, organ injury is still lacking.

[0004] WO 2004 / 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 by inhibiting the complement system. Small peptides (such as sunflower MASP inhibitor-1 (SFMI-1) and sunflower MASP inhibitor-2 (SFMI-2) and their derivatives) for treating diseases associated with the complement system (primarily the lectin pathway) were first described in WO 2010 / 136831.

[0005] WO 2015 / 054298 discloses methods for protecting vision or reducing vision loss in a subject and inhibiting or reducing photoreceptor cell death in a subject by reducing the activity of MASP-1, MASP-2, or MASP-3. WO 2004 / 106384, WO 2005 / 123128, WO 2007 / 117996, and WO 2014 / 144542 disclose anti-MASP-2 antibodies for treating diseases associated with MASP-2-dependent complement activation.

[0006] The object of the present invention is to provide novel peptides that have an inhibitory effect on the MASP-1 and / or MASP-2 enzymes and possess other beneficial properties that make them suitable as effective and safe alternatives for the prevention and / or treatment of diseases associated with MASP-1 and / or MASP-2 as defined below. Another object is to provide novel peptides that have an improved inhibitory effect on the human MASP-1 and / or MASP-2 enzymes and / or rat MASP-1 and / or MASP-2 enzymes.

[0007] The present invention generally relates to peptides useful as inhibitors of the MASP-1 and / or MASP-2 enzymes and methods of making and using the same.

[0008] In particular, the present invention provides compounds containing a peptide comprising, consisting essentially of, or consisting of the following structural formula (I), or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate, or solvate of a salt that can be isolated and / or purified:

[0009] (X 0 ) p (X 1 ) q (X 2 ) r X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X11 X 12 (X 13 ) s (X 14 ) t (X 15 ) u (I),

[0010] in

[0011] X 0 Representing a group of formula (IIa)

[0012]

[0013] in

[0014] * marks the bond to the terminal amino group of the adjacent amino acid,

[0015] A is a bond or C1-C6-alkylene, wherein one CH2 group in C1-C6-alkylene may be replaced by -O- or -S-, and wherein C1-C6-alkylene is up to trisubstituted, identically or differently, by radicals selected from the group consisting of hydroxy, methoxy, ethoxy, carboxyl, amino and halogen,

[0016] B is absent, aryl, heteroaryl, C3-C8-cycloalkyl or C3-C7-heterocycloalkyl,

[0017] wherein aryl, heteroaryl, C3-C8-cycloalkyl and C3-C7-heterocycloalkyl may be up to three times substituted, identically or differently, by radicals selected from the group consisting of C1-C4-alkyl, hydroxy, methoxy, ethoxy, carbonyl, carboxyl, amino and halogen, and

[0018] R 1 is hydrogen, halogen, amino, hydroxyl or C1-C 20 -alkyl, wherein C1-C 20 - alkyl is up to three times substituted, identically or differently, by radicals selected from hydroxy, carboxyl, amino and halogen,

[0019] p represents an integer 0 or 1,

[0020] X 1 represents any natural amino acid or non-natural amino acid, and any natural amino acid and / or non-natural amino acid may be in D- or L-stereoconfiguration,

[0021] And when p is 0 and q is not 0, X 1 The terminal amino group is unsubstituted, acetylated or replaced by a C1-C 20 - alkyl mono- or di-substituted,

[0022] q represents an integer from 0 to 5,

[0023] X 2 represents a natural amino acid selected from I, L, M, V and A, or a non-natural amino acid selected from the group 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,

[0024] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0025] And when p and q are both 0 and r is 1, X 2 The terminal amino group is unsubstituted, acetylated or replaced by a C1-C 20 - alkyl mono- or di-substituted,

[0026] r represents an integer 0 or 1,

[0027] X 3 represents a natural amino acid C, or an unnatural amino acid selected from L-penicillamine (Pen) and LN-methylcysteine ​​((N-Me)C),

[0028] When p, q, and r are all 0, X 3 The terminal amino group is unsubstituted, acetylated or replaced by a C1-C 20 - alkyl mono- or di-substituted,

[0029] X 4 represents a natural amino acid selected from S, C, T, R or K, or an unnatural amino acid selected from allo-L-threonine (allo-T), L-homoserine (hSer) and L-ornithine (Orn),

[0030] X 5 represents the natural amino acid R or N(5)-methyl-L-arginine ((Me)R),

[0031] X 6 represents a natural amino acid selected from S, C or T, or selected from allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap),

[0032] X 7represents a natural amino acid selected from L, F or N or a non-natural amino acid selected from the following: 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-di-fluoro)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-trifluorobutyric acid, L-2-amino-4-cyanobutyric acid (Cnba), 4-fluoro-leucine ((4-fluoro)L), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-( 1-Methylcyclopropyl)propionic acid, L-tert-butylglycine ((tBu)G), 3-(trimethylsilyl)-L-alanine, 2,5-difluoro-L-phenylalanine, 2-amino-7-(tert-butyloxy)-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-fluoro-L-phenylalanine ((2-fluoro)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropionic acid, (2S)-3-(3-cyanophenyl)-2-aminopropionic acid and (2S)-3-(indol-4-yl)-2-(amino)propionic acid,

[0033] X 8 represents a natural amino acid P, or an unnatural amino acid selected from the group consisting of (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, L-hydroxyproline (Hyp), (3S)-morpholine-3-carboxylic acid (morpholine-3-carboxyl), L-homoproline (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]aza-9-carboxylic acid, trans-4-fluoroproline ((trans-4-fluoro)P) and (1R,2S,5S)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid,

[0034] X 9represents a natural amino acid P, or an unnatural amino acid selected from the following: 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), L-hydroxyproline (Hyp), (2S,4S)-4-trifluoromethyl-tetrahydropyrrole-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-trifluorobutyric 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-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S)-2-amino-4,4,4-trifluorobutyric acid, (2S,3aS,6aS)-octahydrocyclopenta]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),

[0035] X 10 represents a natural amino acid I, or an unnatural amino acid selected from the group 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,

[0036] X 11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0037] X 12represents a natural amino acid I, or a non-natural amino acid selected from the group 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),

[0038] When u, t, and s are all 0, X 12 The terminal carboxyl group is unsubstituted or amidated,

[0039] X 13 represents a natural amino acid selected from P, A, S, T, G, D, E, Q or N, or an unnatural amino acid selected from the following: 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 (β-homo-P),

[0040] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0041] If u and t are both 0 and s is not 0, then X 13 The terminal carboxyl group is not substituted or amidated,

[0042] s represents an integer from 0 to 3,

[0043] X 14 represents any natural amino acid or non-natural amino acid, and any natural amino acid and / or non-natural amino acid may be in D- or L-stereoconfiguration,

[0044] If u is 0 and t is not 0, then X 14 The terminal carboxyl group is not substituted or amidated,

[0045] t represents an integer from 0 to 4,

[0046] X 15Representing a group of formula (IIb)

[0047]

[0048] in

[0049] * marks the bond connecting to the terminal carboxyl group of the adjacent amino acid,

[0050] A 1 is a bond or C1-C6-alkylene, wherein one CH2 group in the C1-C6-alkylene may be replaced by -O- or -S-, and wherein the C1-C6-alkylene is up to trisubstituted, identically or differently, by radicals selected from the group consisting of hydroxy, methoxy, ethoxy, carboxyl, amino and halogen,

[0051] B 1 is absent, aryl, heteroaryl, C3-C8-cycloalkyl or C3-C7-heterocycloalkyl,

[0052] wherein aryl, heteroaryl, C3-C8-cycloalkyl and C3-C7-heterocycloalkyl may be up to three times substituted, identically or differently, by radicals selected from the group consisting of C1-C4-alkyl, hydroxy, methoxy, ethoxy, carbonyl, carboxyl, amino and halogen, and

[0053] R 2 is hydrogen, halogen, amino, hydroxyl or C1-C 20 -alkyl, wherein C1-C 20 - alkyl is up to three times substituted, identically or differently, by radicals selected from hydroxy, carboxyl, amino and halogen,

[0054] u represents an integer 0 or 1,

[0055] The condition is X 1 To X 14 At least one of them is an unnatural amino acid.

[0056] The present invention further provides a compound comprising a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt of the following formula (II):

[0057] (X 1 ) q (X 2 ) r X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 (X 13 )s (X 14 ) t (II),

[0058] in

[0059] X 1 represents a natural amino acid or an unnatural amino acid,

[0060] q represents an integer 0 or 1,

[0061] X 2 represents natural amino acid I,

[0062] r represents an integer 0 or 1,

[0063] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0064] X 4 Represents the natural amino acid S,

[0065] X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R),

[0066] X 6 Represents the natural amino acid S,

[0067] X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A),

[0068] X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H),

[0069] 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),

[0070] X 10 represents natural amino acid I,

[0071] X 11 represents an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen)

[0072] X 12 represents natural amino acid I,

[0073] X 13 Represents the natural amino acid P,

[0074] s represents an integer 0 or 1,

[0075] X 14 represents a natural amino acid selected from D, Q and E,

[0076] t represents an integer 0 or 1,

[0077] The N-terminus of the peptide is not substituted, acetylated or C1-C 20 - alkyl mono- or di-substituted,

[0078] wherein the C-terminus of the peptide is not substituted or amidated, and

[0079] Wherein the peptide is cyclized, preferably by linking X 3 and X 11 Key connection.

[0080] Unless otherwise defined herein, the 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 and techniques related to 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.

[0081] Throughout this specification, the word "comprise" or variations thereof such as "comprises" or "comprising" will be understood to imply the inclusion of the stated integer (or component) or group of integers (or components), but not to exclude the inclusion of any other integer (or component) or group of integers (or components). Unless the context clearly dictates otherwise, the singular forms "a", "an" and "the" include the plural forms. The terms "including" and "containing" are used to mean "including but not limited to", and the two expressions can be used interchangeably. In particular, the expression "peptide-containing compound" means a compound that contains a defined peptide sequence and may optionally contain other chemical groups or substituents covalently bound to the peptide, such as amino acids, fatty acids, chemical groups to enhance the pharmacodynamic or pharmacokinetic properties of the peptide or any other chemical group. It will also be understood that the expression "peptide-containing compound" explicitly includes the defined peptide sequence without any other chemical groups or substituents covalently bound to the peptide.

[0082] As used herein, unless otherwise indicated, the following terms have the meanings ascribed to them. "Consisting essentially of" is understood to mean that a 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.

[0083] The terms "protein," "polypeptide," and "peptide" are used interchangeably to refer generally to a sequence of two or more amino acids linked together, preferably by peptide (amide) bonds. 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 also be understood that the terms "protein," "polypeptide," and "peptide" do not indicate a specific length of an amino acid polymer, nor are they intended to imply or distinguish whether a polypeptide is produced using recombinant techniques, chemical or enzymatic synthesis, or is naturally occurring. It should also be understood that, under the definitions of this application, a peptide may include one or more moieties that are not amino acids. These moieties are preferably present at the N- and C-termini of the peptide.

[0084] As used herein, the term "amino acid" or "any amino acid" refers to an organic compound containing an amine (-NH2) and a carboxyl (-COOH) functional group and a side chain, and refers to any and all amino acids, including naturally occurring amino acids (e.g., α-L-amino acids), non-natural amino acids, modified amino acids, and non-natural amino acids. "Natural amino acids" include those amino acids found in nature, such as the 23 amino acids that combine into peptide chains to form the building blocks of a large number of proteins. These are primarily L stereoisomers, although some D-amino acids are found in bacterial envelopes and some antibiotics. Table 2 lists the 20 proteinogenic natural amino acids in the standard genetic code. "Non-standard" natural amino acids are pyrrolysine (present in methanogens and other eukaryotic organisms), selenocystine (present in many non-eukaryotic organisms as well as most eukaryotic organisms), and N-formylmethionine (encoded by the start codon AUG in bacteria, mitochondria, and chloroplasts).

[0085] "Unnatural" or "non-natural" amino acids are naturally occurring or chemically synthesized non-proteinogenic amino acids (i.e., those not naturally encoded or found in the genetic code). More than 140 natural amino acids are known, and thousands of combinations are possible. Examples of "unnatural" amino acids include 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 core amino acids, diamino acids, D-amino acids and N-methyl amino 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, multiple groups, or chemical moieties that are not naturally present in amino acids. Preferred unnatural amino acids according to the present invention are listed in Table 1. Table 1 shows unnatural amino acids as D- and / or L-stereoisomers, however, preferred unnatural amino acids of the present invention are the unnatural D- and L-stereoisomers listed in Table 1.

[0086] Table 1: Preferred unnatural amino acids

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093] More preferred unnatural amino acids are selected from 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, 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-methylbutanoic acid, 4-(aminomethyl)benzoic acid, L-2-aminobutyric acid (Abu), 1-aminocyclobutane-1-carboxylic acid (ACBA), 6-aminohexane Acid (Ahx), 2-aminoisobutyric acid (Aib), L-2-thiophenylalanine (β-2-thiophenylalanine), β-alanine (β-A), β-proline (β-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), γ-aminobutyric acid (γ-Abu), L-3-methylhistidine (3-MeH), L-dihydroorotic acid (Hoo), L-norleucine (Nle), N-methyl-L-proline The present invention also contains amino acids ((N-Me)P), L-norvaline (Nva), L-ornithine (Orn), L-homoproline (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), LN-methylcysteine ​​((N-Me)C), N(5)-methyl-L-arginine ((Me)R), L-penicillamine (Pen) and tranexamic acid (tranexamic acid).

[0094] The most preferred unnatural amino acids are selected from 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).

[0095] It is also understood that the peptides of the present invention may contain one or more chemical groups that are not amino acids within the meaning of the present invention. These chemical groups may be present at the N-terminus and / or C-terminus of the peptide and are 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 by peptide (amide) bonds. Typically, peptides are formed by linking the α-amino and carboxyl groups of α-amino acids, which are then linked by α-peptide bonds. According to the present invention, peptide bonds can be formed by any carboxyl and amino groups present in the corresponding natural or non-natural amino acids. For example, α-amino acids that contain a second amino group in addition to the α-amino group (e.g., L-lysine) or α-amino acids that contain a second carboxyl group in addition to the α-carboxyl group (e.g., L-aspartic acid and L-glutamic acid) can be linked through other amino or carboxyl groups.

[0096] According to the understanding of those skilled in the art, the peptide sequences disclosed herein represent amino acid sequences connected by α-peptide bonds. Amino acids connected by peptide bonds (non-α-peptide bonds) are marked with "*". The "*" is located on the left or right side of the amino acid to indicate whether the other amino group or other carboxyl group of the amino acid is used to connect the peptide bond of the adjacent amino acid (e.g., (*L), (E*), (*Dap), etc.).

[0097] As understood by those skilled in the art, the peptide sequences disclosed herein are shown from left to right, with the left end of the sequence being the "N-terminus" ("amino terminus," "N-terminus") of the peptide and the right end of the sequence being the "C-terminus" ("carboxyl terminus," "C-terminus") of the peptide. The term N-terminus (amino terminus, N-terminus)" applies regardless of whether the peptide actually contains an amino group at the N-terminus. The term C-terminus (carboxyl terminus, C-terminus) applies regardless of whether the peptide actually contains a carboxyl group at the C-terminus. The term "terminal amino group" refers to any amino group present at the N-terminus. The term "terminal carboxyl group" refers to any carboxyl group present at the C-terminus.

[0098] According to the present invention, in the case where p represents 1, the N-terminus can be represented by X 0 Alternatively, in the case where q represents at least 1 and p represents 0, the N-terminus may be formed by X 1Alternatively, when r represents 1, p and q both represent 0, the N-terminus may be formed by X 2 In the case where p, q and r all represent 0, in the case of a linear peptide, the N-terminus is represented by X 3 If the peptide of the present invention is connected by X 3 and X 11 If the peptide is cyclized by a bond and p, q and r all represent 0, the peptide does not comprise an N-terminus.

[0099] According to the present invention, in the case where u represents 1, the C-terminus may be represented by X 15 Alternatively, in the case where t represents an integer of at least 1 and u represents 0, the C-terminus may be formed by X 14 Alternatively, when s represents 1 and both t and u represent 0, the C-terminus may be formed by X 13 In the case where s, t and u all represent 0, the C-terminus can be formed by X 12 form.

[0100] In the present invention, the names of naturally occurring and non-naturally occurring aminoacyl residues used herein preferably follow the naming conventions recommended by the IUPAC Commission on the Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on Biochemical Nomenclature, as listed in Nomenclature of α-Amino Acids (Recommendations, 1974), Biochemistry, 14(2), (1975).

[0101] In this specification, naturally occurring proteinogenic amino acids are generally referred to by their conventional single-letter abbreviations. Alternatively, they may be represented by their three-letter abbreviations (e.g., particularly in sequence listings) or their full names, as shown in Table 2 below:

[0102] Table 2: Standard abbreviations for naturally occurring amino acids

[0103]

[0104] For non-proteinogenic or non-naturally occurring amino acids, unless referred to by their full name (e.g., ornithine, etc.), three to six letter codes are generally used for their residues, including those abbreviations shown in the list of abbreviations below (Table 3).

[0105] As used herein, the term "L-amino acid" refers to the "L" isomeric form of an amino acid. Conversely, the term "D-amino acid" refers to the "D" isomeric form of an amino acid. It is also conventional to use capital letters such as Ala / A, Arg / R, etc. to represent L-amino acids and lowercase letters such as ala / a, arg / r, etc. to represent D-amino acids.

[0106] As shown in Table 2 above, the three-letter codes (code) (i.e., Ala, Arg, Asn, etc.) in the table, and as commonly used in this specification, unless otherwise explicitly stated, generally should include D- and L- forms as well as high- (homo-) and normal- (nor) forms. The prefix "normal" refers to a structural analogue that can be derived from the parent compound by removing a carbon atom and accompanying hydrogen atoms. The prefix "high" represents the next higher member in the homology series. Reference to a specific isomeric form will be represented by the capitalized prefix L- or D- as described above (e.g., D-Arg, L-Arg, etc.). Therefore, specific references to high-form or normal-form will be clearly represented by respective prefixes (e.g., high-Arg, high-R, normal-Arg, normal-R, high-Cys, high-C, etc.).

[0107] The term "C1-C6-alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5 or 6 carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 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 or an isomer thereof. In particular, the radical has 1, 2, 3 or 4 carbon atoms ("C1-C4-alkyl"), for example methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl or tert-butyl, more particularly 1, 2 or 3 carbon atoms ("C1-C3-alkyl"), for example methyl, ethyl, n-propyl or isopropyl. Particularly preferred are methyl, ethyl and n-propyl. Most preferred is methyl.

[0108] The term "C1-C 20 "-alkyl" means a straight-chain or branched saturated monovalent hydrocarbon radical having 1 to 20 carbon atoms, for example 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.

[0109] The term "C1-C4-alkylene" refers to a straight or branched hydrocarbon bridge having 1 to 4 carbon atoms, for example, methylene, ethylene, propylene, α-methylethylene, β-methylethylene, α-ethylethylene, β-ethylethylene, butylene, α-methylpropylene, β-methylpropylene and γ-methylpropylene.

[0110] The term "C1-C6-alkylene" refers to a straight-chain or branched hydrocarbon bridge having 1 to 6 carbon atoms, for example 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.

[0111] 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 is, for example, a monocyclic hydrocarbon ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, a bicyclic hydrocarbon ring, such as bicyclo[4.2.0]octyl or octahydropentalenyl, or a bridged or caged saturated ring group, such as norbornane or adamantane and cubane.

[0112] The term "C3-C7-heterocycloalkyl" refers to a saturated heterocycle with 4, 5, 6 or 7 ring heteroatoms containing one or two identical or different ring heteroatoms from the groups of N, O and S, and the heterocycloalkyl group may be connected to the rest of the molecule via any one carbon atom or nitrogen atom (if present). The C3-C7-heterocycloalkyl group may be, but is not limited to, a four-membered ring such as azetidinyl, oxetanyl or thietanyl; or a five-membered ring such as tetrahydrofuranyl, 1,3-dioxolane, thiolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, 1,1-dioxathiolanyl, 1,2-oxazolidinyl, 1,3-oxazolidinyl or 1,3-thienyl; or a six-membered ring such as tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, thiazolin ... oxazinyl, 1,4-oxazepan ...

[0113] The term "aryl" refers to an unsaturated or partially unsaturated ring having 6 to 10 carbon atoms. Preferred aryl groups are phenyl and naphthyl.

[0114] The term "heteroaryl" refers to a monovalent mono-, bi- 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 and optionally one, two or three further ring heteroatoms from the group consisting of N, O and / or S, and which is attached via a ring carbon atom or, if valence permits, via a ring nitrogen atom. The heteroaryl group can be a 5-membered heteroaryl group, such as thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl or tetrazolyl; or a 6-membered heteroaryl group, such as pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl; or a tricyclic heteroaryl group, such as carbazolyl, acridinyl or phenazinyl; or a 9-membered heteroaryl group, such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzothiazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl, indolindenyl or purinyl; or a 10-membered heteroaryl group, such as quinolinyl, quinazolinyl, isoquinolinyl, cinnamyl, phthalazinyl, quinoxalinyl or pteridinyl.

[0115] In general, unless otherwise indicated, a heteroaryl or heteroarylene group includes all possible isomeric forms thereof, for example, tautomers and positional isomers with respect to the point of attachment to the rest of the molecule. Thus, for some illustrative, non-limiting examples, the term pyridyl includes pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl; or the term thienyl includes thien-2-yl and thien-3-yl.

[0116] Included in the sequences disclosed herein are sequences in which a "-OH" moiety or a "-NH2" moiety is incorporated at the carboxyl terminus (C-terminus) of the sequence. The "-OH" or "NH2" moiety at the C-terminus of the sequence represents a hydroxyl or amino group, corresponding to the presence of a carboxyl or acylamino group (-(C=O)-NH2) at the C-terminus, respectively. In each sequence of the present invention, the C-terminal "-OH" moiety can be replaced by a C-terminal "-NH2" moiety, also referred to herein as an "amidated C-terminus," and vice versa. However, among the alternatives, the C-terminal "-OH" moiety is preferred.

[0117] The term "acetylated" (also abbreviated as "Ac") refers to the acetyl protection of the N-terminal portion of the peptide by acetylation of the N-terminus of the peptide (the N-terminus of the peptide is acetylated).

[0118] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of formula (I), wherein

[0119] X 0 is a chemical group selected from the following: (1S,2S,4S)-bicyclo[2.2.1]hept-5-en-2-ylacetic acid, (2,4-dioxoimidazolin-1-yl)acetic acid, 2-(thiomorpholino)acetic acid ((2-thiomorpholino)acetyl), 2-(N-isopropyl-N-methylamino)acetic acid, (S)-3-methylpentanoic acid ((S)-3-methylpentanoic acid), (S)-3-methylpentanoic acid), 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetyl, 2-(diethylamino)acetic acid (2-(diethylamino)acetyl), 2-(morpholino)acetic acid (2-(morpholino)acetyl), 2-(N-methyl-N-cyclopropylamino)acetyl (2-(N-methyl-N-cyclopropylamino)acetyl), 2-(piperidinyl)acetyl (2-(piperidinyl)acetyl), 2-(pyrrolidinyl)acetyl (2-(pyrrolidinyl)acetyl), 2-hydroxyacetic acid (2-hydroxyacetyl), 2-hydroxyisobutyric acid (2-hydroxyisobutyric acid), 3-(aminomethyl)benzoic acid, 3-methoxypropionic acid, 4-(aminomethyl)benzoic acid, 4-methylheptanoic acid (4 -methylvaleric acid), 5-chlorothiophene-carboxylic acid, 1-(aminomethyl)-cyclopropyl-1-carboxylic acid (ACMP), adipic acid, (S)-azetidine, benzoic acid (benzoic acid), 4-(3,5-dimethyl-1,2-oxazol-4-yl)-L-phenylalanine, cyclobutanecarboxylic acid (cyclobutanecarboxylic acid), 2-(cyclobutyl)acetic acid (cyclobutylacetic acid), cyclobutylacetic acid (cyclobutylacetic acid), cyclohexylacetic acid (cyclohexylacetic acid), cyclohexanecarboxylic acid (cyclohexanecarboxylic acid), cyclopentanecarboxylic acid, cyclopentylacetic acid (cyclopentylacetic acid), cyclopropanecarboxylic acid (cyclobutylacetic acid), propanecarboxylic acid), cyclopropylcarboxylic acid, D-(+)-biotin (Boitin), fumaric acid, 3-phenylpropionic acid (3-phenylpropionic acid), isobutyric acid, isovaleric acid (isovaleric acid), L-(+)-lactic acid (L-(+)-lactic acid) ((lactic acid)(Lactic)), phenylacetic acid, piperidin-4-ylacetic acid, pivalic acid (trimethylacetic acid), suberic acid, tert-butylacetic acid, tetrahydropyran-4-acetic acid, tetrahydro-2H-pyran-3-ylacetic acid and trans-2-(3-((tert-butoxy)carbonylamino)cyclohexyl)acetic acid),

[0120] p represents an integer 0 or 1,

[0121] X 1represents a natural amino acid selected from the group consisting of A, F, G, H, I, K, L, P, R, S, T, V, W and Y or a non-natural amino acid selected from the group consisting of 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, 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 4-aminomethylbenzoic acid, L-2-aminobutyric acid (Abu), 1-aminocyclobutane-1-carboxylic acid (ACBA), 6-aminohexanoic acid (Ahx), 2-aminoisobutyric acid (Aib), L-2-thienylalanine (β-2-thienylalanine), β-alanine (β-A), β-proline (β-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diamino Propionic acid (Dap), γ-aminobutyric acid (γ-Abu), L-3-methylhistidine (3-Me)H), L-dihydroorotic acid (Hoo), L-norleucine (Nle), N-methyl-L-proline ((N-Me)P), L-norvaline (Nva), L-ornithine (Orn), L-piperidinic acid (Pip), (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid and tranexamic acid (tranexamic acid),

[0122] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0123] q represents an integer from 0 to 5,

[0124] X 2 represents a natural amino acid I or a non-natural amino acid selected from the group 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-methyl-heptanoic acid, (2S,3S)-2-[2-oxopiperazin-1-yl]-3-methylpentanoic acid and (2S,3S)-2-[(3S)-2-oxopiperazin-1-yl]-3-methylpentanoic acid,

[0125] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0126] r represents an integer 0 or 1,

[0127] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0128] X 4 represents a natural amino acid selected from S, C, T, R and K,

[0129] X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R),

[0130] X 6 represents the natural amino acid S, or an unnatural amino acid selected from allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap),

[0131] X 7 represents a natural amino acid selected from L and N, or a non-natural amino acid selected from the following: 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-adipic acid (AAD), (2S)-2-amino-4,4,4-trifluorobutyric acid, L-2-amino-4-cyanobutyric acid (Cnba), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propionic acid, (2S)-3-(2,3-difluorophenyl)-2-aminopropionic 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,

[0132] X 8represents a natural amino acid P, or an unnatural amino acid selected from the group 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-homoproline (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]aza-9-carboxylic acid,

[0133] X 9 represents a natural amino acid P, or a non-natural amino acid selected from the following: 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, (2S)-2-amino-4,4,4-trifluorobutyric 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),

[0134] X 10 represents a natural amino acid I, or a non-natural amino acid selected from the group consisting of (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, 3-chlorophenylglycine ((3-chloro-Ph)G), (S)-2-amino-3-ethyl-heptanoic acid, allo-L-isoleucine (allo-I), L-cyclohexylglycine (Chg), L-cyclopentylglycine (Cpg), L-cyclobutylglycine and L-norvaline (Nva),

[0135] X 11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0136] X12 represents the natural amino acid I, or a non-natural amino acid selected from the group 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,

[0137] When u, t, and s are all 0, X 12 The terminal carboxyl group is unsubstituted or amidated,

[0138] X 13 represents a natural amino acid selected from P, A and D or an unnatural amino acid selected from the group 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),

[0139] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0140] If u and t are both 0 and s is not 0, then X 13 The terminal carboxyl group is not substituted or amidated,

[0141] s represents an integer from 0 to 3,

[0142] X 14 represents a natural amino acid selected from the group consisting of D, E, G, K, N, P and Q or a non-natural amino acid selected from the group 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 (β-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic acid),

[0143] And when u is 0 and t is not 0, X 14 The terminal carboxyl group is not substituted or amidated,

[0144] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0145] t represents an integer from 0 to 4,

[0146] X 15 is a chemical group selected from the group 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)propionic acid, (S)-pyrrolidine-3-carboxylic acid, 5-azaspiro[2.4]heptane-1-carboxylic acid and (2S)-3-(triazol-1-yl)-2-(amino)propionic acid,

[0147] u represents an integer 0 or 1,

[0148] With the proviso that the peptide comprises at least one unnatural amino acid.

[0149] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of formula (I), wherein

[0150] X 0 is a chemical group selected from the group consisting of (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, (2-thiomorpholino)acetic acid, (N-isopropyl-N-methylamino)acetic acid, 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetic acid, 2-(diethylamino)acetic acid, 2-(morpholino)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, benzoic acid, cyclobutylacetic acid, cyclopropylacetic acid, phenylacetic acid, piperidin-4-ylacetic acid, tetrahydro-2H-pyran-3-ylacetic acid, tranexamic acid, and trans-2-(3-((tert-butoxy)carbonylamino)cyclohexyl)acetic acid,

[0151] p represents an integer 0 or 1,

[0152] X 1represents a natural amino acid selected from the group consisting of A, F, G, H, I, K, L, P, R, S, T, V, W and Y or a non-natural amino acid 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, β-2 -thiophene alanine, (S)-pyrrolidine-2-carboxylic acid (β-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), γ-aminobutyric acid (γ-Abu), L-3-methylhistidine (H(3-Me)), L-dihydroorotic acid (Hoo), L-norleucine (Nle), L-norvaline (Nva), L-ornithine (Orn), L-homoproline (Pip), N-methyl-L-proline ((N-Me)P), tranexamic acid (tranexamic acid), (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid and (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid,

[0153] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0154] q represents an integer from 0 to 5,

[0155] X 2 represents a natural amino acid I or an unnatural amino acid (Abu) selected from LN-methylisoleucine ((N-Me)I), L-cyclobutylalanine (Cba), L-norvaline (Nva) and L-2-aminobutyric acid,

[0156] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0157] r represents an integer 0 or 1,

[0158] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0159] X 4 represents a natural amino acid selected from S and T,

[0160] X 5Represents the natural amino acid R,

[0161] X 6 represents the natural amino acid S or the unnatural amino acid allo-L-threonine (allo-T),

[0162] X 7 represents a natural amino acid selected from N and L, or a non-natural amino acid selected from the group 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)propionic acid and 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), 3-(trimethylsilyl)-L-alanine,

[0163] X 8 represents a natural amino acid P, or an unnatural amino acid selected from (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]aza-9-carboxylic acid,

[0164] X 9 represents a natural amino acid P, or a non-natural amino acid selected from the following: 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),

[0165] X 10 represents a natural amino acid I, or an unnatural amino acid selected from (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, (S)-2-amino-3-ethyl-pentanoic acid, L-cyclohexylglycine (Chg) and L-cyclopentylglycine (Cpg),

[0166] X11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0167] X 12 represents the natural amino acid I, or an unnatural amino acid selected from 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,

[0168] When u, t, and s are all 0, X 12 The terminal carboxyl group is not substituted or amidated,

[0169] X 13 represents a natural amino acid selected from P and D, or an unnatural amino acid selected from 2-aminoisobutyric acid (Aib), 2-methyl-L-proline (2-Me)P and trans-4-fluoroproline ((trans-4-fluoro)P),

[0170] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0171] If u and t are both 0 and s is not 0, then X 13 The terminal carboxyl group is not substituted or amidated,

[0172] s represents an integer from 0 to 3,

[0173] X 14 represents a natural amino acid selected from D, Q, N, E and P or an unnatural amino acid selected from the group consisting of 3-carboxyphenylalanine ((3-carboxy)F), N-methyl-glycine ((N-Me)G), (2S)-pyrrolidin-2-yl acetic acid (β-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic acid),

[0174] Each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a D-stereoisomer.

[0175] And when u is 0 and t is not 0, X 14 The terminal carboxyl group is not substituted or amidated,

[0176] t represents an integer from 0 to 4,

[0177] X 15is a chemical group selected from the group consisting of (1R,3S)-3-(amino)cyclopentanecarboxylic acid, (1S,3R)-3-(amino)cyclopentanecarboxylic acid, (R)-4-amino-6-methylheptanecarboxylic acid, (S)-(1-homoproline-3-yl)-acetic acid, (S)-3-(1-pyrrolidin-2-yl)-propionic acid, (S)-3-(2H-tetrazol-5)-yl)propionic acid, (S)-pyrrolidine-3-carboxylic acid, 5-azaspiro[2.4]heptane-1-carboxylic acid and (2S)-3-(triazol-1-yl)-2-(amino)propionic acid,

[0178] u represents an integer 0 or 1,

[0179] With the proviso that the peptide comprises at least one unnatural amino acid.

[0180] According to another embodiment, the present invention provides a compound comprising a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of formula (I), wherein

[0181] p represents the integer 0,

[0182] X 1 represents a natural amino acid selected from A and G or a non-natural amino acid selected from the group 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),

[0183] q represents an integer 0 or 1,

[0184] X 2 represents natural amino acid I,

[0185] r represents an integer 0 or 1,

[0186] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0187] X 4 Represents the natural amino acid S,

[0188] X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R),

[0189] X 6 Represents the natural amino acid S,

[0190] X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A),

[0191] X 8 Represents the natural amino acid P,

[0192] 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),

[0193] X 10 represents natural amino acid I,

[0194] X 11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0195] X 12 represents natural amino acid I,

[0196] If u, t, and s are all 0, then X 12 The terminal carboxyl group is not substituted or amidated,

[0197] X 13 Represents the natural amino acid P,

[0198] If u and t are both 0 and s is not 0, then X 13 The terminal carboxyl group is not substituted or amidated,

[0199] s represents an integer 0 or 1,

[0200] X 14 represents a natural amino acid selected from D, Q and E,

[0201] And when u is 0 and t is not 0, X 14 The terminal carboxyl group is not substituted or amidated,

[0202] t represents an integer 0 or 1,

[0203] u represents the integer 0,

[0204] With the proviso that the peptide comprises at least one unnatural amino acid.

[0205] According to another embodiment, the present invention provides a compound comprising a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt of the following formula (II):

[0206] (X 1 ) q (X 2 ) r X 3 X 4 X 5 X 6 X7 X 8 X 9 X 10 X 11 X 12 (X 13 ) s (X 14 ) t (II),

[0207] in

[0208] X 1 represents a natural amino acid selected from A and G, or an unnatural amino acid selected from N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn),

[0209] q represents an integer 0 or 1,

[0210] X 2 represents natural amino acid I,

[0211] r represents an integer 0 or 1,

[0212] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0213] X 4 Represents the natural amino acid S,

[0214] X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R),

[0215] X 6 Represents the natural amino acid S,

[0216] X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A),

[0217] X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H),

[0218] 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),

[0219] X 10 represents natural amino acid I,

[0220] X 11represents an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0221] X 12 represents natural amino acid I,

[0222] X 13 Represents the natural amino acid P,

[0223] s represents an integer 0 or 1,

[0224] X 14 represents a natural amino acid selected from D, Q and E,

[0225] t represents an integer 0 or 1,

[0226] The N-terminus of the peptide is not substituted, acetylated or C1-C 20 - alkyl mono- or di-substituted,

[0227] wherein the C-terminus of the peptide is not substituted or amidated, and

[0228] The peptide is linked by X 3 and X 11 cyclization of the bond.

[0229] According to another embodiment, the present invention provides a compound comprising a peptide of formula (II) or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, wherein

[0230] X 1 represents a natural amino acid or an unnatural amino acid,

[0231] q represents an integer 0 or 1,

[0232] X 2 represents natural amino acid I,

[0233] r represents an integer 0 or 1,

[0234] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0235] X 4 Represents the natural amino acid S,

[0236] X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R),

[0237] X 6 Represents the natural amino acid S,

[0238] X 7represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A),

[0239] X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H),

[0240] X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic),

[0241] X 10 represents natural amino acid I,

[0242] X 11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0243] X 12 represents natural amino acid I,

[0244] X 13 Represents the natural amino acid P,

[0245] s represents an integer 0 or 1,

[0246] X 14 represents a natural amino acid selected from D, Q and E,

[0247] t represents an integer 0 or 1,

[0248] The N-terminus of the peptide is not substituted, acetylated or C1-C 20 - alkyl mono- or di-substituted,

[0249] wherein the C-terminus of the peptide is not substituted or amidated, and

[0250] The peptide is linked by X 3 and X 11 cyclization of the bond.

[0251] According to another embodiment, the present invention provides a compound comprising a peptide of formula (II) or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, wherein

[0252] X 1 represents a natural amino acid selected from A and G, or an unnatural amino acid selected from N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn),

[0253] q represents an integer 0 or 1,

[0254] X 2 represents natural amino acid I,

[0255] r represents an integer 0 or 1,

[0256] X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen),

[0257] X 4 Represents the natural amino acid S,

[0258] X 5 represents the natural amino acid R or the unnatural amino acid N(5)-methyl-L-arginine ((Me)R),

[0259] X 6 Represents the natural amino acid S,

[0260] X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A),

[0261] X 8 represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H),

[0262] X 9 represents the unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic),

[0263] X 10 represents natural amino acid I,

[0264] X 11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen),

[0265] X 12 represents natural amino acid I,

[0266] X 13 Represents the natural amino acid P,

[0267] s represents an integer 0 or 1,

[0268] X 14 represents a natural amino acid selected from D, Q and E,

[0269] t represents an integer 0 or 1,

[0270] Wherein the N-terminus of the peptide is not substituted or acetylated,

[0271] wherein the C-terminus of the peptide is not substituted or amidated, and

[0272] The peptide is linked by X 3 and X 11 cyclization of the bond.

[0273] In particular, the present invention provides a compound containing a peptide of formula (I) or a compound containing a peptide of formula (II).

[0274] According to the present invention, X 1 To X 14 The following definitions apply to peptides of formula (I) and (II), position X 0 and X 15 The definitions of apply to the peptides of formula (I).

[0275] According to the present invention, X 0 (if present) may be a non-amino acid chemical group not according to the definition herein.

[0276] According to another embodiment of the present invention, X 0represents a chemical group selected from the group consisting of 2-cyanobenzoic acid, (1S,2S,4S)-bicyclo[2.2.1]hept-5-en-2-yl acetic acid, (2,4-dioxoimidazolin-1-yl)acetic acid, 2-(thiomorpholino)acetic acid ((2-thiomorpholino)acetyl), 2-(N-isopropyl-N-methylamino)acetic acid, (S)-3-methylpentanoic acid ((S)-3-methylpentanoic acid), 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetyl, 2-( acetic acid (2-(diethylamino)acetyl), 2-(morpholino)acetic acid (2-(morpholino)acetyl), 2-(N-methyl-N-cyclopropylamino)acetyl (2-(N-methyl-N-cyclopropylamino)acetyl), 2-(piperidinyl)acetyl (2-(piperidinyl)acetyl), 2-(pyrrolidinyl)acetyl (2-(pyrrolidinyl)acetyl), 2-hydroxyacetic acid (2-hydroxyacetyl), 2-hydroxyisobutyric acid (2-hydroxyisobutyric acid), 3-(aminomethyl)benzoic acid, 3-methoxypropionic acid, 4-( Aminomethyl)benzoic acid, 4-methylpentanoic acid (4-methylpentanoic acid), 5-chlorothiophenecarboxylic acid, 1-(aminomethyl)-cyclopropyl-1-carboxylic acid (ACMP), adipic acid, (S)-azetidine, benzoic acid (benzoic acid), 4-(3,5-dimethyl-1,2-oxazol-4-yl)-L-phenylalanine, cyclobutanecarboxylic acid (cyclobutanecarboxylic acid), 2-(cyclobutyl)acetic acid (cyclobutylacetic acid), cyclobutylacetic acid (cyclobutylacetic acid), cyclohexylacetic acid (cyclohexylacetic acid), cyclohexanecarboxylic acid (cyclohexanecarboxylic acid) , cyclopentanecarboxylic acid, cyclopentylacetic acid (cyclopentylacetic acid), cyclopropanecarboxylic acid (cyclopropanecarboxylic acid), cyclopropylcarboxylic acid, D-(+)-biotin, fumaric acid, 3-phenylpropionic acid (phenylpropionic acid), isobutyric acid, isovaleric acid (isovaleric acid), L-(+)-lactic acid (lactic acid), phenylacetic acid, piperidin-4-ylacetic acid, pivalic acid (trimethylacetic acid), suberic acid, tert-butylacetic acid, tetrahydropyranyl-4-acetic acid, tetrahydro-2H-pyran-3-ylacetic acid and trans-2-(3-((tert-butoxy)carbonylamino)cyclohexyl)acetic acid).

[0277] According to another embodiment of the present invention, X 0represents a chemical group selected from the group consisting of 2-cyanobenzoic acid, (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid, (2-thiomorpholino)acetic acid, (N-isopropyl-N-methylamino)acetic acid, 2-(3-pyridyl)acetic acid, 2-(cyclohexylamino)acetic acid, 2-(diethylamino)acetic acid, 2-(morpholino)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, benzoic acid, cyclobutylacetic acid, cyclopropylacetic acid, phenylacetic acid, piperidin-4-ylacetic acid, tetrahydro-2H-pyran-3-ylacetic acid, tranexamic acid and trans-2-(3-((tert-butoxy)carbonylamino)cyclohexyl)acetic acid.

[0278] According to another embodiment of the present invention, X 0 is a chemical group selected from an unnatural amino acid as defined above, 2-cyanobenzoic acid, a substituted benzoic acid or phenylacetic acid.

[0279] According to another embodiment of the present invention, X 0 It's Hoo, ODD or Ahx.

[0280] According to another embodiment of the present invention, p represents the integer 1.

[0281] According to another embodiment of the present invention, p represents the integer 0.

[0282] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 1 represents any natural amino acid or unnatural amino acid, preferably selected from the unnatural amino acids listed in Figure 1, and each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a stereoisomer with a D-stereoconfiguration.

[0283] According to another embodiment of the present invention, X 1represents a natural amino acid selected from the group consisting of A, F, G, H, I, K, L, P, R, S, T, V, W and Y, or a non-natural amino acid selected from the group consisting of 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, 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-methylbutanoic acid, 4-(aminomethyl)benzoic acid Acid, L-2-aminobutyric acid (Abu), 1-aminocyclobutane-1-carboxylic acid (ACBA), 6-aminohexanoic acid (Ahx), 2-aminoisobutyric acid (Aib), L-2-thiophenylalanine (β-2-thiophenylalanine), β-alanine (β-A), β-proline (β-P), L-citrulline (Cit), L-2,4-diaminobutyric acid (Dab), L-2,3-diaminopropionic acid (Dap), γ-aminobutyric acid (γ-Abu), L-3-methylhistidine (3-Me)H), L-dihydroorotic acid (Hoo), L-norleucine (Nle), N-methyl-L-proline ((N-Me)P), L-norvaline (Nva), L-ornithine (Orn), L-homoproline (Pip), (2S)-2[(amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid and tranexamic acid (tranexamic acid), and each L-stereoconfiguration of natural amino acids and / or unnatural amino acids can be replaced by a stereoisomer with a D-stereoconfiguration.

[0284] According to another embodiment of the present invention, X l represents a natural amino acid selected from A and G, or an unnatural amino acid selected from N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn), and each natural amino acid and / or unnatural amino acid with an L-stereoconfiguration can be replaced by a stereoisomer with a D-stereoconfiguration.

[0285] According to another embodiment of the present invention, X lrepresents a natural amino acid selected from A and G, or an unnatural amino acid selected from N-methyl-L-alanine (N-Me)A, N-methyl-glycine ((N-Me)G), L-norleucine (Nle), L-norvaline (Nva) and L-ornithine (Orn).

[0286] According to another embodiment of the present invention, X 1 represents the natural amino acid A or the unnatural amino acid N-methyl-glycine ((N-Me)G).

[0287] According to another embodiment of the present invention, when p is 0 and q is not 0, X 1 The terminal amino group is unsubstituted, acetylated or mono- or disubstituted by C1-C4-alkyl, preferably C1-C 20 -Alkyl monosubstituted.

[0288] According to another embodiment of the present invention, X 1 is methylated.

[0289] According to another embodiment of the present invention, X 1 Acetylated.

[0290] According to another embodiment of the present invention, q represents the integer 0.

[0291] According to another embodiment of the present invention, q represents the integer 1.

[0292] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 2 represents a natural amino acid selected from I, L, M, V and A, or a non-natural amino acid selected from the following: 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, and each natural amino acid and / or non-natural amino acid in the L-stereoconfiguration may be replaced by a stereoisomer in the D-stereoconfiguration.

[0293] According to another embodiment of the present invention, X2 represents a natural amino acid I or a non-natural amino acid selected from the following: 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, and each L-stereoconfiguration of the natural amino acid and / or non-natural amino acid can be replaced by a stereoisomer with a D-stereoconfiguration.

[0294] According to another embodiment of the present invention, X2 represents a natural amino acid I or a non-natural amino acid selected from LN-methylisoleucine ((N-Me)I), L-cyclobutylalanine (Cba), L-norvaline (Nva) and L-2-aminobutyric acid (Abu), and each L-stereoconfiguration natural amino acid and / or non-natural amino acid can be replaced by a stereoisomer with a D-stereoconfiguration.

[0295] According to another embodiment of the present invention, X 2 Represents natural amino acid I.

[0296] According to another embodiment of the present invention, when p and q are both 0 and r is 1, X 2 The terminal amino group is not substituted, acetylated or replaced by C1-C 20 -alkyl is mono- or disubstituted, preferably monosubstituted by C1-C4-alkyl.

[0297] According to another embodiment of the present invention, r represents the integer 0.

[0298] According to another embodiment of the present invention, r represents the integer 1.

[0299] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 3 represents a natural amino acid C, or an unnatural amino acid selected from L-penicillamine (Pen) and LN-methylcysteine ​​((N-Me)C).

[0300] According to another embodiment of the present invention, X 3 represents the natural amino acid C or the unnatural amino acid L-penicillamine (Pen).

[0301] According to another embodiment of the present invention, X 3 Represents the natural amino acid C.

[0302] According to another embodiment of the present invention, when p, q and r are all 0, X 3 The terminal amino group is unsubstituted, acetylated or mono- or disubstituted by C1-C4-alkyl, preferably C1-C 20 -Alkyl monosubstituted.

[0303] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 4 represents a natural amino acid selected from S, C, T, R or K, or an unnatural amino acid (allo-T) selected from allo-L-threonine, L-homoserine (hSer) and L-ornithine (Orn).

[0304] According to another embodiment of the present invention, X 4 represents a natural amino acid selected from S, C, T, R and K.

[0305] According to another embodiment of the present invention, X 4 represents a natural amino acid selected from S or T.

[0306] According to another embodiment of the present invention, X 4 Represents the natural amino acid S.

[0307] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can 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).

[0308] According to another 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).

[0309] According to another embodiment of the present invention, X 5 Represents the natural amino acid R.

[0310] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 6 represents a natural amino acid selected from S, C or T, or an unnatural amino acid selected from allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap).

[0311] According to another embodiment of the present invention, X 6 represents the natural amino acid S or an unnatural amino acid selected from allo-L-threonine (allo-T) and L-2,3-diaminopropionic acid (Dap).

[0312] According to another embodiment of the present invention, X 6 represents the natural amino acid S or the unnatural amino acid allo-L-threonine (allo-T).

[0313] According to another embodiment of the present invention, X 6 Represents the natural amino acid S.

[0314] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 7represents a natural amino acid selected from L, F or N or a non-natural amino acid selected from the following: 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 amino acid ((2-bromo) F), (S)-2-(amino)-1,6-hexanedioic acid (AAD), (2S)-2-amino-4,4,4-trifluorobutyric acid, L-2-amino-4-cyanobutyric acid (Cnba), 4-fluoro-leucine ((4-fluoro) L), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propionic acid, L-tert-butylglycine ((tB u)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, L-trifluoromethyl Alanine, L-difluoromethylalanine, 2-fluoro-L-phenylalanine ((2-Fluoro)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropionic acid, (2S)-3-(3-cyanophenyl)-2-aminopropionic 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)propionic acid.

[0315] According to another embodiment of the present invention, X 7represents a natural amino acid selected from L and N, or a non-natural amino acid selected from the following: 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-adipic acid (AAD), (2S)-2-amino-4,4,4-trifluorobutyric acid, L-2-amino-4-cyanobutyric acid (Cnba), (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propionic acid, (2S)-3-(2,3-difluorophenyl)-2-aminopropionic 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.

[0316] According to another embodiment of the present invention, X 7 represents a natural amino acid selected from N or L, or a non-natural amino acid selected from the following: 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)propionic acid and 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic), 3-(trimethylsilyl)-L-alanine.

[0317] According to another embodiment of the present invention, X 7 represents the natural amino acid L or the unnatural amino acid L-tert-butylalanine ((tBu)A).

[0318] According to another embodiment of the present invention, X 7 represents the unnatural amino acid L-tert-butylalanine ((tBu)A).

[0319] According to another embodiment of the present invention, X 7 Represents the natural amino acid L.

[0320] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 8 represents a natural amino acid P, or a non-natural amino acid selected from the following: 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-homoproline ( 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.

[0321] According to another embodiment of the present invention, X 8 represents a natural amino acid P, or a non-natural amino acid selected from the following: L-proline (3,4-2H), (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, trans-4-fluoroproline ((cis-4-fluoro)P), L-hydroxyproline (Hyp), (3S)-morpholine-3-carboxylic acid (morpholine-3-carboxylic acid), L-homoproline (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]aza-9-carboxylic acid.

[0322] According to another embodiment of the present invention, X 8 represents a natural amino acid P, or an unnatural amino acid selected from 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]aza-9-carboxylic acid.

[0323] According to another embodiment of the present invention, X 8 Represents the natural amino acid P or the unnatural amino acid L-proline (3,4-2H).

[0324] According to another embodiment of the present invention, X 8 Represents the natural amino acid P.

[0325] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 9 represents a natural amino acid P, or an unnatural amino acid selected from the following: 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-trifluorobutyric 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-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S)-2-amino-4,4,4-trifluorobutyric acid, (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).

[0326] According to another embodiment of the present invention, X 9represents a natural amino acid P, or a non-natural amino acid selected from the following: 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, (2S)-2-amino-4,4,4-trifluorobutyric 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).

[0327] According to another embodiment of the present invention, X 9 represents a natural amino acid P, or a non-natural amino acid selected from the following: 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).

[0328] According to another embodiment of the present invention, X 9 represents natural amino acid P or unnatural amino acid 2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (Oic)o

[0329] According to another 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)

[0330] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 10 represents a natural amino acid I, or a non-natural amino acid selected from the group 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.

[0331] According to another embodiment of the present invention, X 10 represents the natural amino acid I, or a non-natural amino acid selected from the group consisting of: (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, 3-chlorophenylglycine ((3-chloro-Ph)G), (S)-2-amino-3-ethyl-heptanoic acid, allo-L-isoleucine (allo-I), L-cyclohexylglycine (Chg), L-cyclopentylglycine (Cpg), L-cyclobutylglycine, and L-norvaline (Nva).

[0332] According to another embodiment of the present invention, X 10 represents a natural amino acid I, or a non-natural amino acid selected from (2S)-2-(amino)-2-[(1S,3R)-3-hydroxycyclohexyl]acetic acid, (S)-2-amino-3-ethyl-pentanoic acid, L-cyclohexylglycine (Chg) and L-cyclopentylglycine (Cpg).

[0333] According to another embodiment of the present invention, X 10 Represents natural amino acid I.

[0334] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 11 represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0335] According to another embodiment of the present invention, X 11represents a natural amino acid C, or an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0336] According to another embodiment of the present invention, X 11 represents an unnatural amino acid selected from LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0337] According to another embodiment of the present invention, X 11 represents the unnatural amino acid L-penicillamine (Pen).

[0338] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can 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).

[0339] According to another 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).

[0340] According to another 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 LN-methylcysteine ​​((N-Me)C) and L-penicillamine (Pen).

[0341] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 12represents the natural amino acid I, or a non-natural amino acid selected from the group consisting of allo-L-isoleucine (allo-I), (S)-2-amino-2-cyclobutylacetic acid (Cbg), (2S,3S)-2-((amino)methyl)-3-methylheptanoic acid, L-phenylglycine (Phg), 2-[(1S,2S)-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, 2-methyl-D-isoleucine, L-norvaline (Nva), L-2-aminobutyric acid (Abu), L-tert-butylglycine, and aminoisobutyric acid (Aib).

[0342] According to another embodiment of the present invention, X 12 represents the natural amino acid I, or a non-natural amino acid selected from the group 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-isoleucine and (2S,3S)-2-((amino)methyl)-3-methylpentanoic acid.

[0343] According to another embodiment of the present invention, X 12 represents the natural amino acid I, or an unnatural amino acid selected from 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.

[0344] According to another embodiment of the present invention, X 12 Represents natural amino acid I.

[0345] According to another embodiment of the present invention, when u, t and s are all 0, X 12 The terminal carboxyl group is not substituted or amidated.

[0346] According to another embodiment, the present invention provides compounds containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of Formula (I) or Formula (II), wherein X 13represents a natural amino acid selected from P, A, S, T, G, D, E, Q or N or a non-natural amino acid selected from the following: 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), Formula -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 (β-homo-P), and each L-stereoconfiguration of natural amino acids and / or non-natural amino acids can be replaced by a stereoisomer with a D-stereoconfiguration.

[0347] According to another embodiment of the present invention, X 13 represents a natural amino acid selected from P, A and D or an unnatural amino acid selected from 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), and each natural amino acid and / or unnatural amino acid of the L-stereoconfiguration can be replaced by a stereoisomer of the D-stereoconfiguration.

[0348] According to another embodiment of the present invention, X 13 represents a natural amino acid selected from P or D or a non-natural amino acid selected from 2-aminoisobutyric acid (Aib), 2-methyl-L-proline (2-Me) P and trans-4-fluoroproline ((trans-4-fluoro) P), and each natural amino acid and / or non-natural amino acid of the L-stereoconfiguration can be replaced by a stereoisomer of the D-stereoconfiguration.

[0349] According to another embodiment of the present invention, X 13 Represents the natural amino acid P.

[0350] According to another embodiment of the present invention, s represents the integer 0.

[0351] According to another embodiment of the present invention, s represents the integer 1.

[0352] According to another embodiment of the present invention, when u and t are both 0 and s is not 0, X 13 The terminal carboxyl group is unsubstituted or amidated.

[0353] According to another embodiment of the present invention, when t is 0 and s is 1, X 13 The terminal carboxyl group is not substituted or amidated.

[0354] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein X 14 represents any natural amino acid or unnatural amino acid, and any natural amino acid and / or unnatural amino acid may be in the D- or L-stereoconfiguration.

[0355] According to another embodiment of the present invention, X 14 represents a natural amino acid selected from D, E, G, K, N, P and Q or an unnatural amino acid selected from the following: 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-yl acetic acid (β-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic acid), and each natural amino acid and / or unnatural amino acid of the L-stereoconfiguration can be replaced by a stereoisomer of the D-stereoconfiguration.

[0356] According to another embodiment of the present invention, X 14 represents a natural amino acid selected from D, Q, N, E and P or an unnatural amino acid selected from the following: 3-carboxyphenylalanine ((3-carboxy)F), N-methyl-glycine ((N-Me)G), (2S)-pyrrolidin-2-yl acetic acid (β-homo-P), L-2,3-diaminopropionic acid (Dap), L-ornithine (Orn) and tranexamic acid (tranexamic acid), and each L-stereoconfiguration natural amino acid and / or unnatural amino acid can be replaced by a stereoisomer with a D-stereoconfiguration.

[0357] According to another embodiment of the present invention, X 14 represents a natural amino acid selected from D, Q or I.

[0358] According to another embodiment of the present invention, X 14 Represents the natural amino acid D.

[0359] According to another embodiment of the present invention, X 14 The C-terminal "-OH" moiety at is replaced by a C-terminal "-NH2" moiety.

[0360] According to another embodiment of the present invention, q represents the integer 0.

[0361] According to another embodiment of the present invention, q represents the integer 1.

[0362] According to another embodiment of the present invention, in the case where s and t represent the integer 1, X 14 The terminal carboxyl group is not substituted or amidated.

[0363] According to another embodiment of the present invention, in the case where s and t represent the integer 1, X 14 The terminal carboxyl group is unsubstituted.

[0364] According to the present invention, X 15 (if present) may be a chemical group other than an amino acid.

[0365] According to another embodiment of the present invention, X 15 is a chemical group selected from the following: -NH2, -NH-C1-C6-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)propionic acid, (S)-pyrrolidine-3-carboxylic acid, 5-azaspiro[2.4]heptane-1-carboxylic acid and (2S)-3-(triazol-1-yl)-2-(amino)propionic acid.

[0366] According to another embodiment of the present invention, X 15 is a chemical group selected from -NH2 or -NH-C1-C6-alkyl (preferably -NH2).

[0367] According to another embodiment of the present invention, q represents the integer 0.

[0368] According to another embodiment of the present invention, q represents the integer 1.

[0369] According to another embodiment of the present invention, the N-terminus of the peptide is not substituted, acetylated or replaced by C1-C 20 -Alkyl is mono- or di-substituted.

[0370] According to another embodiment of the invention, the N-terminus of the peptide is unsubstituted, acetylated or mono- or disubstituted by C1-C4-alkyl.

[0371] According to another embodiment of the present invention, the N-terminus of the peptide is not substituted or acetylated.

[0372] According to another embodiment of the present invention, the C-terminus is not substituted or amidated.

[0373] According to another embodiment of the present invention, the N-terminus and the C-terminus are unsubstituted.

[0374] According to another embodiment, the present invention provides a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of Formula (I) or Formula (II), which is selected from Examples 29, 44, 45, 67, 75, 109, 166, 185, 443 and 466 or is selected from Examples 29, 44, 45, 67, 75, 109, 166, 185, 443, 466 and 499.

[0375] The peptides of formula (I) or (II) may be linear or cyclized. As used herein, the term "cyclization" refers to a reaction in which one portion of a polypeptide molecule (e.g., Cys, (N-Me)Cys, or Pen) is linked to another portion of a polypeptide molecule (e.g., another Cys, (N-Me)Cys, or Pen) to form a closed ring, for example, by forming a disulfide bond (-SS-) or other similar bonds, such as 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 a carbon-carbon bond (e.g., (-CH2-CH2-)). According to the present invention, X of the peptide 3 and X 11 Linkages between positions are preferred. In the peptide sequences of the present invention, an amino acid followed by a "+" refers to a cyclic peptide, in which an amino acid is linked to another amino acid followed by a "+" via a disulfide bond, forming a closed ring. Alternatively, two linked chemical groups in a polypeptide molecule can be represented by a connecting line.

[0376] According to another embodiment, the present invention provides a compound containing a peptide or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of Formula (I) or Formula (II), wherein the peptide is cyclized.

[0377] According to another embodiment of the present invention, the peptide of formula (I) or formula (II) is linked to X 3 and X 11 cyclization of the bond.

[0378] According to another embodiment of the present invention, the peptide of formula (I) or formula (II) is linked to X3 and X 11 The bond of 3 and X 11 Connected through a linker of formula -SS-, -CH2-S-, -S-CH2-, -CH2-CH2-, -S-(CH2)2-, -(CH2)2-S- or -CH2-S-CH2-.

[0379] According to another embodiment of the present invention, the peptide of formula (I) or formula (II) is linked to X 3 and X 11 The bond of 3 and X 11 The linkage is via a disulfide bond of formula -SS-.

[0380] The peptide of the present invention can be replaced with a suitable water-soluble polymer characterized by a repeating unit. Suitable polymers can be selected from polyalkoxy polymers, hyaluronic acid and derivatives thereof, polyvinyl alcohol, polyoxazoline, polyanhydrides, poly(orthoesters), polycarbonates, polyurethanes, polyacrylic acids, polyacrylamides, polyacrylates, polymethacrylates, polyorganophosphazenes, polysiloxanes, polyvinyl pyrrolidone, polycyanoacrylates and polyesters.

[0381] The peptide of the present invention can be substituted by at least one polyethylene group (PEG group). At least one PEG group is preferably combined with the N-terminal and / or C-terminal. The PEG group can be combined with any suitable functional group of the chemical group and / or amino acid of the peptide, such as hydroxyl, carboxyl, amino, thiol, preferably amino or carboxyl. Preferably, the peptide of the present invention contains a PEG group combined with the N-terminal or a PEG group combined with the C-terminal. More preferably, the PEG group is combined with the N-terminal or C-terminal through an amide bond.

[0382] The concept of PEGylation of peptides to improve their solubility, reduce immunogenicity, improve stability and / or increase half-life by reducing renal clearance has been well known since the early 1980s (Caliceti P., Veronese FM, Adv. Drug Deliv. Rev. 2003, 55, 1261-1277). This has been successfully used for several drugs, but in many instances, PEGylation reduces the efficacy of the drug substance to the point that the concept is no longer applicable (T. Peleg-Shulman et al., J. Med. Chem., 2004, 47, 4897-4904).

[0383] The PEG group of the present invention is any group comprising at least two ethylene oxide units to form an ethylene oxide oligomer or polymer. The PEG group is covalently coupled (PEGylated) to the peptide of the present invention, which is then referred to as a PEGylated peptide. Generally, this type of modification of molecules is well known in the art.

[0384] The PEG group can be composed of an interconnecting portion, a polymer portion, and an end group. The interconnecting portion can be composed of C1-C 20 alkyl groups, which are optionally interrupted or terminated by heteroatoms or functional groups selected from -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, wherein R X is hydrogen or C1-C6-alkyl, which is optionally interrupted or terminated by one or more of the above atoms or groups, which further has hydrogen as a terminal atom. The polymer portion consists of at least two ethylene oxide units, which are optionally interrupted or terminated by one or more heteroatoms or functional groups selected from -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, wherein R X is hydrogen or C1-C6-alkyl. The terminal group may be composed of heteroatoms or functional groups selected from 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 terminal groups are COOH and C(O)NH2.

[0385] Examples of PEG spacers of 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), and the like.

[0386] It is noted that the PEG groups defined herein may be labeled by different names by commercial suppliers, but this should not preclude the existence of identical compounds of this invention with different names.

[0387] According to another embodiment, the present invention provides a compound containing a peptide or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of Formula (I) or Formula (II), wherein the peptide is substituted with at least one PEG group.

[0388] According to another embodiment of the invention, the peptide is substituted with a PEG group.

[0389] According to another embodiment of the invention, the peptide is substituted with a PEG group, wherein the PEG group is attached to the C-terminus, preferably via an amide bond.

[0390] According to another embodiment of the present invention, the peptide is substituted with at least one PEG group, wherein at least one or more PEG groups are groups of formula (IIIa)

[0391]

[0392] in

[0393] * marks the connection to nitrogen or oxygen atoms,

[0394] D is C1-C4-alkylene,

[0395] Y is selected from hydroxy, methoxy, ethoxy, carboxyl, carboxamide or amino, and

[0396] n represents an integer from 2 to 15,

[0397] or a group of formula (IIIb),

[0398]

[0399] in

[0400] * Marks the combination with carbonyl group,

[0401] D 1 is C1-C4-alkylene,

[0402] Y 1 is selected from hydroxy, methoxy, ethoxy, carboxyl, carboxamide or amino, and

[0403] m represents an integer from 2 to 15, or TTDS.

[0404] According to another embodiment of the present invention, the peptide is substituted with at least one PEG group, and at least one or more PEG groups are selected from 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.

[0405] The peptides of the present invention may comprise at least one C8-C 20 Fatty acids. Typically, such fatty acids may be branched or cyclic. 20 The fatty acid is preferably bound to the N-terminus and / or the C-terminus. 20 The fatty acid may be bound to any suitable functional group of the peptide chemical group and / or amino acid, such as a hydroxyl group, a carboxyl group, an amino group, a thiol group, preferably an amino group or a carboxyl group. Preferably, the peptide of the present invention contains a C8-C 20 Fatty acids or C8-C 20 More preferably, a C8-C 20 The fatty acid is bound to the N-terminus or C-terminus via an amide bond. 0 or X 15 The fatty acid side chain formed is a fatty acid with a C8 residue, more preferably a fatty acid with a C 12 Fatty acids, more preferably ≥C 14 Further preferably, X 0 or X 15 The fatty acid side chain formed is C 12 -C 18 Fatty acids, preferably C 12 -C 16 Fatty acids, or C 14 -C 18 Fatty acids, or C 14 -C 16 Fatty acids. Most preferably C16 Fatty acids such as palmitic acid (palmitoyl, Palm) and C 18 Fatty acids such as 1,18-octadecanedioic acid (ODD).

[0406] It is also understood that the N-terminal or C-terminal moiety may be a bond (eg, a covalent bond), particularly where the amino terminus or carboxyl terminus is bound to a linker or another chemical moiety.

[0407] Chemical groups, unnatural amino acids, or moieties may be abbreviated herein as shown in Table 3.

[0408] Table 3: Abbreviations / expressions and terms used for chemical groups, unnatural amino acids, or other parts of the sequence

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418] The term "mimetic" used together with some amino acids in the definitions of several parts of the peptides according to formula (I) or formula (II) of the present invention means the corresponding amino acid mimics, such as arginine mimics, isoleucine mimics or proline mimics. Typically, a "protein mimic" means a molecule, such as a peptide, a modified peptide or any other molecule that biologically simulates the function or activity of some other protein. In the context of using the term "mimetic" in connection with a certain amino acid, the term "mimetic" similarly means any other amino acid, amino acid analog, amino acid derivative, amino acid conjugate, etc. that biologically simulates the function or activity of the corresponding amino acid.

[0419] The proline mimetics of the present invention particularly include (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] aza-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-azasilone-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 prolines.

[0420] Isoleucine mimetics of the present invention particularly include (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-methylheptanoic acid, Phg, 2-[(1S,2S))-1-(amino)-2-methylbutyl]-1,3-oxazole-4-carboxylic acid, 2-methyl-D-alloisoleucine, Nva, Abu or Ala.

[0421] The leucine mimetics of the present invention particularly include (tBu)A, (2-chloro)F, (2-bromo)F, AAD, (2S)-2-amino-4,4,4-trifluorobutyric acid, Cnba, (4-fluoro)L, (S)-(trifluoromethyl)-L-cysteine, (2S)-2-amino-3-(1-methylcyclopropyl)propionic acid, Gly(tBu), 3-(trimethylsilyl)-L-alanine, 2,5-difluoro-L-phenylalanine, 2-amino-7-(tert-butyloxy)-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-aminopropionic acid, (2S)-3-(3-cyanophenyl))-2-aminopropionic 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)propionic acid.

[0422] The present invention also includes analogs and derivatives of the peptides. The term "analogs" or "derivatives" of the peptides or amino acid sequences according to the present invention particularly includes any amino acid sequence having at least 80% or at least 85%, preferably at least 90%, more preferably at least 95% sequence identity, even more preferably at least 99% identity to the sequence, and the same or comparable properties or activities. Sequence identity can be determined by conventional techniques, such as visual comparison or by any computer tool commonly used in the art. Examples include the BLAST program using default parameters.

[0423] Analogs or derivatives of the peptides or amino acid sequences of the present invention may be produced by mutations or variations of the peptide sequences of the present invention, including deletions or insertions of one or more amino acids or substitutions 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 relative to the amino acid sequence.

[0424] As used herein, the term "conservative substitution" means that one or more amino acids are replaced with another biologically similar residue. Examples include replacing 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. See, for example, the scheme in Table 4 below, where conservative substitutions of amino acids are grouped by physicochemical properties. I: neutral, hydrophilic; II: acids and amides; III: basic; IV: hydrophobic; V: aromatic, bulky amino acids; VI: neutral or hydrophobic; VII: acidic; VIII: polar.

[0425] Table 4: Amino acids grouped according to their physicochemical properties

[0426] I II III IV V VI VII VIII Ala Asn His Met Phe Ala Glu Met Ser Asp Arg Leu Tyr Leu Asp Ser Thr Glu Lvs Ile Trp Ile Thr Pro Gln Val Pro Cys Gly Cys Gly Asn Val Gln

[0427] The peptide analog or derivative may also comprise one or more other modifications, such as conjugation to another compound to form an amino acid conjugate.

[0428] Alternatively or additionally, such modifications may result from conjugation to the side chains of one or more amino acid residues in the peptides of the invention, for example a ligand having a chemical group X as defined above. 0 In some embodiments, the present invention provides a kind of peptide of the present invention.For example ...

[0429] Unless otherwise indicated, all peptides of the present invention are TFA salts. The present invention also includes pharmaceutically acceptable salts and salt-free forms of the peptides defined herein. A pharmaceutically acceptable salt represents a salt or zwitterionic form of a peptide or compound of the present invention that is water-soluble, oil-soluble, or dispersible, suitable for treating a disease without excessive 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 an 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, mesitylenesulfonate, methanesulfonate, naphthalenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, trichloroacetate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, p-toluenesulfonate, and undecanoate. Preferred acid addition salts include trifluoroacetate, formate, hydrochloride, and acetate.

[0430] In addition, the amino groups in the compounds of the present invention may be quaternized by 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 and phenethyl bromides. 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. Pharmaceutically acceptable salts may suitably be salts selected from, for example, acid addition salts and basic salts. Examples of acid addition salts include chloride salts, citrates and acetates.

[0431] Examples of basic salts include those wherein the cation is selected from alkali metal cations such as sodium or potassium, alkaline earth metal cations such as calcium or magnesium, and substituted ammonium ions such as N(R 1 )(R 2 )(R 3 )(R 4 ) + salts of type ions, where R 1 、R 2 、R 3 and R 4 independently of one another generally represent hydrogen, optionally substituted C 1-6 -alkyl or optionally substituted C 2-6 -alkenyl. Related C 1-6 Examples of -alkyl groups include methyl, ethyl, 1-propyl and 2-propyl. 2-6 Examples of the -alkenyl group include vinyl, 1-propenyl and 2-propenyl. Among them, salts having a cation selected from sodium, potassium and calcium are preferred.

[0432] Other examples of pharmaceutically acceptable salts are described in "Remington's Pharmaceutical Sciences", 17th edition, Alfonso R. Gennaro (ed.), Mark Publishing Company, Easton, PA, USA, 1985 (and updated editions thereof), and in "Encyclopaedia of Pharmaceutical Technology", 3rd edition, James Swarbrick (ed.), Informa Healthcare USA (Inc.), NY, USA, 2007. In addition, for a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002). Other suitable basic 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 hemisulphate and hemicalcium salts.

[0433] The present invention also includes solvates of the peptides defined herein. The term "solvate" refers to a complex having a defined stoichiometry formed between a solute (e.g., a peptide of the present invention or a pharmaceutically acceptable salt thereof) and a solvent. The solvent in the conjugate may be, for example, water, ethanol, or another pharmaceutically acceptable, typically small molecule, organic substance, such as, but not limited to, acetic acid or lactic acid. When the solvent in question is water, such solvates are generally referred to as hydrates.

[0434] The compounds of the present invention have useful pharmacological properties and can be used for the prevention and treatment of disorders in humans and animals.

[0435] In the context of the present invention, the term "treatment" or "treat" includes inhibiting, delaying, curbing, ameliorating, attenuating, limiting, reducing, preventing, reversing or curing a disease, condition, disorder, injury or impairment of health, the development, course or progression of such states and / or symptoms of such states. Herein, the term "therapy" is understood to be synonymous with the term "treatment".

[0436] In the context of the present invention, the terms "prevention," "prophylaxis," or "precaution" are used synonymously and refer to avoiding or reducing the risk of acquiring, contracting, suffering from, or having a disease, condition, disorder, injury, or impairment of health, the development or progression of such states or symptoms of such states.

[0437] Treatment or prevention of a disease, condition, disorder, injury or impairment of health may be carried out partially or completely.

[0438] 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.

[0439] The compounds of the present invention can therefore be used in medicaments for the treatment and / or prevention of cardiovascular and cardiopulmonary disorders and their sequelae, for example inflammatory heart diseases, myocarditis, endocarditis, pericarditis, rheumatic fever with and without heart involvement, acute rheumatic pericarditis, acute rheumatic endocarditis, acute rheumatic myocarditis, chronic rheumatic heart diseases with or without endocarditis, valvulitis, pericarditis, ischemic heart diseases 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 tachycardia, atrial fibrillation, atrial flutter, ventricular fibrillation, ventricular flutter, ventricular tachyarrhythmias, torsades de pointes tachycardia, atrial and ventricular extrasystoles, AV-junctional extrasystoles, sick sinus syndrome syndrome), stroke caused by occlusion and stenosis of cerebral arteries (e.g., cerebral infarction following thromboembolic, atherosclerotic, infectious and inflammatory vasculopathy); treatment and / or prevention of stroke caused by intracerebral or intracranial hemorrhage, peripheral ischemic tissue damage (e.g., atherosclerotic gangrene), phlebitis and thrombophlebitis caused by diseases of the arteries, arterioles and capillaries (e.g., thromboembolic, atherosclerotic, infectious and inflammatory vasculopathy, deforming or occlusive endarteritis and aneurysmal dissection); prevention of postoperative disorders of the circulatory system, such as systemic inflammatory response syndrome, postoperative vasoplegia, postcardiotomy syndrome, postoperative hypotension and heart failure; prevention and treatment of ischemia-reperfusion injury and organ dysfunction, such as after thrombolytic therapy, percutaneous transluminal angioplasty (PTA), percutaneous transluminal coronary angioplasty (PTCA), bypass surgery and heart, lung, liver and kidney transplantation; and prevention and treatment of delayed graft function after renal transplantation.

[0440] By preventing MASP-mediated end-organ damage, the compounds of the invention are also useful in treating shock, such as cardiogenic shock, septic shock, and anaphylactic shock.

[0441] Furthermore, the compounds of the present invention have anti-inflammatory effects and can therefore be used as anti-inflammatory agents for the treatment and / or prevention of sepsis (SIRS), multiple organ failure (MODS, MOF), inflammatory conditions of the kidneys, chronic intestinal inflammation (IBD, Crohn's disease, UC), pancreatitis, peritonitis, rheumatoid diseases, inflammatory skin diseases and inflammatory eye diseases.

[0442] By virtue of their activity profile, the compounds according to 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.

[0443] The compounds of the present invention are particularly useful for treating and / or preventing ischemia and / or reperfusion-related damage to the heart and kidneys, as well as 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.

[0444] The compounds according to the invention can furthermore be used for preventing ischemia- and / or reperfusion-related damage to organs or tissues and can also be used as additives to perfusion and preservation solutions for organs, organ parts, tissues or tissue parts of human or animal origin, in particular in the field of surgical intervention or transplantation medicine.

[0445] Furthermore, the compounds of the invention are suitable for the treatment and / or prevention of diseases of the blood and hematopoietic organs and the immune system, including but not limited to acquired hemolytic anemia, hemolytic-uraemic syndrome, paroxysmal nocturnal hemoglobinuria [Marchiafava-Micheli], coagulation defects, purpura and other bleeding disorders, disseminated intravascular coagulation [defibrosis syndrome], essential (hemorrhagic) thrombocythemia, purpura fulminans, thrombotic thrombocytopenic purpura, Henoch-Schonlein purpura, allergic vasculitis, lymphocytopenia and granulocytosis and sarcoidosis.

[0446] Furthermore, the compounds according to the invention are suitable for the treatment and / or prevention of sequelae of diabetes, for example renal complications of diabetes, diabetic nephropathy, intracapillary glomerulonephrosis, ophthalmological complications of diabetes, diabetic retinopathy, neurological complications, diabetic polyneuropathy and circulatory complications such as microangiopathy and gangrene.

[0447] Furthermore, the compounds according to the invention are suitable for the treatment and / or prophylaxis of inflammatory diseases of the nervous system, for example multiple sclerosis, meningitis and encephalitis, bacterial and viral meningitis and encephalitis, postimmune encephalitis, inflammatory polyneuropathies and polyneuropathies in infectious and parasitic diseases.

[0448] The compounds according to the invention are also suitable for the treatment and / or prevention 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 retinopathy and retinal vascular disease. changes), proliferative retinopathy, macular and posterior pole degeneration, peripheral retinal degeneration, age-related macular degeneration (AMD) (including dry (non-exudative) and wet (exudative, neovascular) AMD), choroidal neovascularization (CNV), choroidal neovascular membrane (CNVM), cystoid macular edema (CME), epiretinal membrane (ERM) and macular perforation, myopia-related choroidal neovascularization, angioids and vascular streaks, retinal detachment, diabetic retinopathy, diabetic macular edema (DME), atrophic and hypertrophic retinal pigment epithelium, retinal vein occlusion, chorioretinal vein occlusion, macular edema, macular edema associated with retinal vein occlusion, postoperative conditions of the eye and adnexa such as keratopathy after cataract surgery.

[0449] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of respiratory diseases, including but not limited to viral, bacterial and fungal pneumonia, radiation pneumonitis, pneumoconiosis, extrinsic allergic alveolitis, airway diseases caused by certain organic dusts (e.g. farmer's lung), bronchitis, pneumonia and pulmonary edema caused by chemicals, gases, fumes and vapors, drug-induced interstitial lung disease, adult respiratory distress syndrome (ARDS) and acute lung injury (ALI), acute pulmonary edema, interstitial lung disease fibrosis, rheumatoid lung disease, respiratory diseases among other diffuse connective tissue diseases, such as those associated with systemic lupus erythematosus, scleroderma and Wegener's granulomatosis.

[0450] 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 serotypes H1N1, H5N1, H7N9) and coronaviruses (e.g. SARS-CoV, the causative agent of severe acute respiratory syndrome (SARS), MERS-CoV, the causative agent of Middle East respiratory syndrome (MERS), and SARS-CoV-2, the causative agent of the COVID-19 pandemic).

[0451] Furthermore, the compounds of the present invention are suitable for the treatment and / or prevention of digestive system diseases, including but not limited to non-infectious enteritis and colitis, such as Crohn's disease and ulcerative colitis, pancreatitis (including alcohol- and drug-induced acute pancreatitis), cholecystitis, inflammatory liver diseases, hepatorenal syndrome, postoperative liver conditions, such as after liver surgery.

[0452] Due to their activity profile, the compounds of the present invention are particularly suitable for the treatment and / or prevention of genitourinary diseases, 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 the case of hemodialysis and hemodiafiltration), cystitis, radiation cystitis, inflammatory diseases of the prostate and endometriosis.

[0453] The compounds of the present invention are also suitable for treating and / or preventing the sequelae of burns and injuries, including but not limited to early complications of trauma, traumatic anuria, crush syndrome, post-crush renal failure, traumatic muscle ischemia, brain trauma, organ damage after exposure to electric current, radiation and extreme ambient air temperature and pressure, organ damage after exposure to smoke, fire and flames, and organ damage after contact with poisonous animals and plants.

[0454] Due to their activity profile, the compounds of the invention are also suitable for the treatment of inflammatory skin diseases such as lupus erythematosus, bullous and acantholytic skin diseases such as pemphigus subtypes, papulosquamous disorders such as psoriasis, dermatitis and eczema, urticaria and erythema.

[0455] According to another embodiment, the present invention provides a compound containing a peptide or its derivative, prodrug, analog or pharmaceutically acceptable salt or solvate that can be isolated and / or purified for use in preventing and / or treating a disease, wherein the peptide comprises, essentially consists of or consists of formula (I) or formula (II).

[0456] According to another embodiment, the present invention provides a compound comprising a peptide comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), or a derivative, prodrug, analog, or pharmaceutically acceptable salt or solvate thereof that can be isolated and / or purified for use in preventing and / or treating a MASP-associated disease.

[0457] According to another embodiment, the present invention provides a compound comprising a peptide that can be isolated and / or purified, or a derivative, prodrug, analog or pharmaceutically acceptable salt or solvate thereof, wherein the peptide comprises, consists essentially of or consists of Formula (I) or Formula (II), and acts as a MASP-1 and / or MASP-2 inhibitory agent and / or inhibits C3 deposition.

[0458] According to another embodiment, the present invention provides a compound containing a peptide or its derivative, prodrug, analog or pharmaceutically acceptable salt or solvate which can be isolated and / or purified, wherein the peptide comprises, essentially consists of or consists of formula (I) or formula (II), and the compound is used to prevent and / or treat cardiovascular and cardiopulmonary diseases, shock, inflammatory diseases, cardiovascular, pulmonary, brain and renal sequelae of sepsis, ischemia and / or reperfusion-related injury, acute kidney injury, transplant protection and delayed graft function, diseases of the blood and hematopoietic organs and the immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, respiratory system diseases, digestive system diseases or genitourinary system and sequelae of burns and injuries.

[0459] According to another embodiment, the present invention provides a compound containing a peptide or its derivative, prodrug, analog or pharmaceutically acceptable salt or solvate which can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II), and the compound is used to prevent and / or treat genitourinary system diseases, including but not limited to acute renal failure, acute kidney injury (AKI), surgery-related AKI, sepsis-related AKI, contrast agent and chemotherapy-induced AKI, renal ischemia and infarction, complications (such as allergies in the case of hemodialysis and hemodiafiltration), cystitis, radiation cystitis, prostatic inflammatory diseases and endometriosis.

[0460] The present invention further relates to a method of treating or ameliorating a MASP-associated disorder as defined above in a subject or patient by administering to said subject or patient in need thereof at least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate, complex or pharmaceutical composition thereof as defined above.

[0461] As used herein, the terms "patient," "subject," or "individual" are used interchangeably and refer to humans or non-human animals. These terms include mammals, such as humans, primates, livestock animals (e.g., bovines, swine), pets (e.g., canines, felines), 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.

[0462] According to the present invention, at least one peptide, derivative or analog as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex 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 invention is intended to describe an amount of the compound of the invention sufficient to treat a MASP-associated disorder as defined herein. In certain embodiments, a therapeutically effective amount will achieve the desired benefit / risk ratio applicable to any medical treatment.

[0463] The present invention particularly encompasses the embodiments wherein the substituents or moieties of the peptides of formula (I) or formula (II) as defined below may independently have the meanings indicated below.

[0464] Peptides or compounds containing a peptide as defined herein, or a derivative or analog thereof, or a pharmaceutically acceptable salt or solvate or complex or pharmaceutical composition (defined below) are also generally referred to hereinafter as "MASP inhibitory peptides of the invention".

[0465] In some embodiments, the MASP-inhibitory peptides of the present invention bind to MASP-1 and / or MASP-2, such as 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, "specific binding" means that a specific binding agent interacts with a given ligand more preferentially than other agents in a sample. For example, a specific binding agent that specifically binds to a given ligand binds to the given ligand under appropriate conditions in an amount or degree that observably exceeds any nonspecific interactions with other components in the sample. Suitable conditions are those that permit 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 given specific binding agents and ligand pairs, but can be readily determined by one skilled 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 of the MASP-inhibitory peptide reference compounds provided herein).

[0466] Thus, the present invention further relates to complexes comprising at least one peptide as defined herein that binds to MASP-1 or MASP-2, or a compound, derivative or analog comprising the peptide.

[0467] In some embodiments, the MASP inhibitory peptides of the invention exhibit specific binding to MASP-1 and / or MASP-2 (particularly human 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 10000% greater than that of a selected MASP inhibitory peptide reference compound.

[0468] In some embodiments, the MASP-inhibitory peptides of the 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 greater, or at least about 10, 20, 50, or 100 times greater than a selected MASP-inhibitory peptide reference compound.

[0469] In some embodiments, the MASP inhibitory peptides of the invention have a 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 10000% greater than that of a selected MASP inhibitory peptide reference compound.

[0470] In some embodiments, the MASP inhibitory peptides of the invention have a binding affinity for MASP-1 and / or MASP-2 that is at least about 1, 2, 3, 4, 5 fold greater, or at least about 10, 20, 50, 100, or 1000 fold greater than a selected MASP inhibitory peptide reference compound.

[0471] In some embodiments, the MASP-inhibitory peptides of the 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 in vitro or in vivo, such as described herein. In some embodiments, the MASP-inhibitory peptides of the invention inhibit the activity of MASP-1 and / or MASP-2 by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10000% of the activity inhibited by a selected MASP-inhibitory peptide reference compound.

[0472] In certain embodiments, the MASP-inhibitory peptides of the 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-fold greater inhibition of MASP-1 and / or MASP-2 than a selected MASP-inhibitory peptide reference compound.

[0473] In another specific embodiment, the inhibitory activity of the MASP-inhibitory peptides of the invention against MASP-1 and / or MASP-2 is determined by measuring their IC values ​​for MASP-1 and / or MASP-2 (e.g., rat human MASP-1 and / or MASP-2). 50 IC for MASP-1 and / or MASP-2 50 This can be accomplished using the biochemical assays described herein. Particularly preferred are MASP inhibitory peptides of the invention that exhibit an IC <RTI ID=0.0>0.0001< / RTI> for MASP-1 and / or MASP-2. 50 <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.

[0474] In some embodiments, the MASP inhibitory peptides of the invention have lower IC values ​​for MASP-1 and / or MASP-2 (e.g., rat or human MASP-1 and / or MASP-2) than selected MASP inhibitory peptide reference compounds. 50 (i.e., higher binding affinity). In some embodiments, the IC values ​​of the MASP inhibitory peptides of the invention in MASP-1 and / or MASP-2 competitive binding assays are 50 Compared with the IC values ​​of selected MASP inhibitory peptide reference compounds 50 At least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000% or 10000% lower.

[0475] In some embodiments, the MASP-inhibitory peptides of the invention exhibit 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 10000% greater inhibition of human MASP-1 and / or MASP-2 activity in vitro compared to a selected MASP-inhibitory peptide reference compound.

[0476] In some embodiments, the MASP-inhibitory peptides of the invention exhibit 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 10000% greater inhibition of human MASP-1 and / or MASP-2 activity in vivo compared to a selected reference MASP-inhibitory peptide compound.

[0477] 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 vitro or in a subject (e.g., a mouse or a human) when administered thereto in a dose-dependent and time-dependent manner (e.g., by a parenteral route, such as by injection, or orally by the pulmonary, nasal, sublingual, lingual, buccal, cutaneous, transdermal, conjunctival, ocular route, or as an implant or stent).

[0478] In some embodiments, the MASP-inhibitory peptides of the invention exhibit inhibition of C3 deposition (e.g., human C3 deposition). In some embodiments, inhibition of C3 deposition is determined by in vitro or in vivo inhibition. In some embodiments, the MASP-inhibitory peptides of the invention inhibit C3 deposition by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 700%, 1000%, or 10000% of the inhibition of C3 deposition by a selected MASP-inhibitory peptide reference compound.

[0479] In certain embodiments, the MASP inhibitory peptides of the invention inhibit C3 deposition 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-fold more than a selected MASP inhibitory peptide reference compound.

[0480] In another specific embodiment, the MASP-1 and / or MASP-2 inhibitory activity of the MASP inhibitory peptides of the invention is determined by measuring their IC50 activity in inhibiting C3 deposition in vitro or in a subject (e.g., a mouse or human). 50 To determine the IC deposited by C3 50 This can be accomplished using the C3 human deposition assay described herein. Particularly preferred are MASP inhibitory peptides of the invention having an IC of C3 deposition of 50 It is <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.

[0481] In some embodiments, the MASP inhibitory peptides of the invention inhibit C3 deposition in vitro by 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 10000% greater than a selected MASP inhibitory peptide reference compound.

[0482] In some embodiments, the MASP inhibitory peptides of the invention exhibit 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 10000% greater inhibition of C3-deposition in vivo than a selected MASP inhibitory peptide reference compound.

[0483] Particularly preferred are peptides of the invention as MASP inhibitory peptides, the activity of which is determined according to at least one of the specific assays and / or in vivo studies of the Examples herein.

[0484] Due to their above-mentioned 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 analogs, derivatives and pharmaceutically acceptable salts or solvates thereof, as well as the above-mentioned complexes) are useful for preventing and / or treating disorders associated with MASP-1 and / or MASP-2.

[0485] According to another embodiment, the invention provides a compound comprising a peptide comprising, consisting essentially of, or consisting of Formula (I) or Formula (II), or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate, or solvate of a salt, which can be isolated and / or purified, that acts as a MASP-1 and / or MASP-2 inhibitory agent and / or inhibits C3 deposition.

[0486] If desired, the compounds of the invention can be used alone or in combination with other active compounds. The present invention also relates to medicaments containing at least one compound of the invention and one or more other active compounds, in particular for the treatment and / or prevention of the above-mentioned diseases. As suitable combined active compounds, we can mention by way of example and preferably:

[0487] compounds that inhibit the degradation of cyclic guanosine monophosphate (cGMP) and / or cyclic adenosine monophosphate (cAMP), for example inhibitors of phosphodiesterase (PDE) 1, 2, 3, 4 and / or 5, in particular PDE 4 inhibitors (for example roflumilast or revamilast) and PDE 5 inhibitors (for example sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, mirodenafil or lodenafil);

[0488] NO-independent but heme-dependent stimulators of guanylate cyclase, in particular riociguat, nelociguat, vericiguat and the compounds described in WO 00 / 06568, WO 00 / 06569, WO 02 / 42301, WO 03 / 095451, WO 2011 / 147809, WO 2012 / 004258, WO 2012 / 028647 and WO 2012 / 059549;

[0489] NO-independent and heme-dependent activators of guanylate cyclase, in particular Runcaciguat, BI703704 and the compounds described in WO 2012 / 139888, WO 2001 / 019780 and WO 2014 / 012934;

[0490] prostacyclin analogs and IP receptor agonists, for example and preferably iloprost, beraprost, treprostinil, epoprostenol, NS-304, selexipag or ralinepag;

[0491] - endothelin receptor antagonists, for example and preferably bosentan, darusentan, ambrisentan, macicentan or sitaxsentan;

[0492] Vasopressin receptor antagonists, such as tolvaptan, conivaptan, and relcovaptan;

[0493] Human neutrophil elastase (HNE) inhibitors, for example and preferably sivelestat or DX-890 (Reltran);

[0494] compounds which inhibit signal transduction cascades, in particular compounds from the group of 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;

[0495] Signal transduction modulators from the group of ASK1 kinase inhibitors, such as selonsertib;

[0496] Rho kinase inhibitors, for example and preferably fasudil, Y-27632, SLx-2119, BF-66851, BF-66852, BF-66853, KI-23095 or BA-1049;

[0497] Active ingredients that reduce the permeability of the vascular wall (edema formation), for example and preferably 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;

[0498] Corticosteroids, such as cortisone, cortisol, prednisolone, methylprednisolone, triamcinolone, or dexamethasone;

[0499] Active ingredients that reduce organ damage under oxidative stress, for example and preferably complement system inhibitors, in particular complement C5a receptor antagonists, anti-C5 antibodies or 5-HT1A receptor agonists;

[0500] Regulators, stimulators and enhancers of the transcription factor Nrf2, such as CXA-10, oltipraz, dimethyl fumarate or bardoxolone;

[0501] Adrenomedullin and adrenomedullin derivatives (e.g., pegylated adrenomedullin) and adrenomedullin stabilizers (e.g., adrecizumab);

[0502] Compounds that inhibit hypoxia-inducible factor prolyl hydroxylases (HIF-PH inhibitors), for example molidustat, vadadustat, roxadustat, daprodustat or desidustat;

[0503] Compounds that inhibit pathways that induce cell death and apoptosis, such as QPI-1002;

[0504] C-Met agonists and hepatocyte growth factor mimetics, such as refanalin;

[0505] Alkaline phosphatase and recombinant alkaline phosphatase;

[0506] Compounds that inhibit inflammatory responses and T-cell proliferation, for example CD28 antagonist compounds such as Reltecimod;

[0507] Compounds that modulate Th17 T cell activation, such as RORc / ROR-γ transcription factor regulators;

[0508] Compounds that antagonize Th17 T cell responses, such as anti-IL-17 antibodies and anti-IL-23 antibodies, for example, ixekizumab, secukinumab, brodalumab, ustekinumab, guselkumab, or PTG-200;

[0509] antithrombotic agents, for example and preferably platelet aggregation inhibitors, anticoagulants or fibrinolytic substances;

[0510] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a platelet aggregation inhibitor, such as for example and preferably aspirin, clopidogrel, ticlopidine or dipyridamole.

[0511] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a thrombin inhibitor, such as for example and preferably ximelagatran, melagatran, dabigatran, bivalirudin or Clexane.

[0512] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a GPIIb / IIIa antagonist, such as for example and preferably tirofiban or abciximab.

[0513] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a factor Xa inhibitor, such as, by way of example and preferably, rivaroxaban, apixaban, fidexaban, razaxaban, fondaparinux, idraparinux, DU-176b, PMD-3112, YM-150, KFA-1982, EMD-503982, MCM-17, MLN-1021, DX 9065a, DPC 906, JTV 803, SSR-126512 or SSR-128428.

[0514] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with heparin or a low molecular weight (LMW) heparin derivative.

[0515] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with direct inhibitors of factor XI, inhibitors of factor XI expression and anti-factor XI antibodies (for example Xisomab 3G3);

[0516] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a mineralocorticoid receptor antagonist, such as for example and preferably spironolactone, eplerenone or finerenone.

[0517] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a diuretic, such as by way of example and preferably furosemide, bumetanide, torsemide, bendroflumethiazide, chlorothiazide, hydrochlorothiazide, hydroflumethiazide, methylchlorothiazide, polythiazide, trichlorothiazide, chlorthalidone, indapamide, metolazone, quinethazone, acetazolamide, dichlorphenamide, methazolamide, glycerol, isosorbide, mannitol, amiloride or triamterene.

[0518] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a PPAR-gamma agonist, such as by way of example and preferably pioglitazone or rosiglitazone.

[0519] In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a PPAR-delta agonist, such as, for example and preferably, GW 501516 or BAY 68-5042.

[0520] According to another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound comprising a peptide (which may be isolated and / or purified) or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, in combination with one or more other active ingredients, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II), and wherein the other active ingredients are selected from the group consisting of phosphodiesterase inhibitors, guanylate cyclase stimulators or activators, IP receptor agonists, mineralocorticoid receptor antagonists, diuretics, PPAR-γ agonists, PPAR-δ agonists, corticosteroids, active ingredients that reduce organ damage under oxidative stress, compounds that inhibit pathways that induce cell death and apoptosis, 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 coagulation factor XI inhibitors.

[0521] The present invention also relates to a kit-of-parts comprising 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, and at least one selected from the group consisting of a reagent, a medical device, instructions or any combination thereof.

[0522] The present invention also 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 delivering said peptide, derivative, analogue or complex or pharmaceutical composition thereof to a subject.

[0523] The pharmaceutical composition, kit-of-parts combination or medical device as defined above is particularly useful for preventing and / or treating a disorder or disease as defined herein.

[0524] The MASP inhibitory peptides of the present invention may be systemically and / or locally active. For this purpose, they can be administered in a suitable manner, for example, by oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, vaginal, cutaneous, transdermal, conjunctival, otic route or as an implant or stent.

[0525] For these administration routes, the compounds according to the invention can be administered in suitable administration forms.

[0526] 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 quickly and / or in a modified manner, such as tablets (uncoated or coated tablets, e.g., enteric or controlled-release coatings with delayed dissolution or insolubility), orally disintegrating tablets, films / thin tablets, films / lyophilized products, capsules (e.g., hard or soft gelatin capsules), sugar-coated tablets, granules, pills, powders, emulsions, suspensions, aerosols, or solutions. The compounds of the present invention can be incorporated into the dosage forms in crystalline and / or amorphous and / or dissolved form.

[0527] Parenteral administration can be achieved by avoiding an absorption step (e.g. intravenously, intraarterially, intracardially, intraspinally or intralumbarly) or by including absorption (e.g. intramuscularly, subcutaneously, intradermally, transdermally or intraperitoneally, intraocularly). Suitable administration forms for parenteral administration are, in particular, injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilizates or sterile powders.

[0528] Examples of suitable other routes of administration are pharmaceutical forms for inhalation [especially powder inhalers, aerosols], nasal drops, nasal solutions, nasal sprays; tablets / films / wafers / capsules for lingual, sublingual or buccal administration; suppositories; eye drops, eye ointments, eye washes, eye inserts, ear drops, ear sprays, ear powders, ear washes, ear plugs; vaginal capsules, topical paints, aqueous suspensions (lotions, mixtures), lipophilic suspensions, emulsions, ointments, creams, transdermal therapeutic systems (e.g. plasters), emulsions, pastes, foams, powders, implants or stents.

[0529] According to another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound containing a peptide (which may be isolated and / or purified) or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, in combination with one or more inert, non-toxic, pharmaceutically suitable excipients, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II).

[0530] The compounds of the invention can be incorporated into the administration forms described. This can be achieved in a manner known per se by mixing with pharmaceutically suitable excipients. Pharmaceutically suitable excipients include, in particular:

[0531] Fillers and carriers (e.g. cellulose, microcrystalline cellulose (e.g. ), lactose, mannitol, starch, calcium phosphate (e.g. )),

[0532] ointment bases (e.g. petrolatum, paraffin, triglycerides, waxes, wool wax, wool alcohol, lanolin, hydrophilic ointments, polyethylene glycol),

[0533] Suppository bases (e.g. polyethylene glycol, cocoa butter, hard fat),

[0534] Solvents (e.g. water, ethanol, isopropanol, glycerol, propylene glycol, medium-chain triglyceride fatty oils, liquid polyethylene glycol, paraffin),

[0535] Surfactants, emulsifiers, dispersants or wetting agents (e.g. sodium lauryl sulfate), lecithin, phospholipids, fatty alcohols (e.g. ), sorbitan fatty acid esters (e.g. ), polyoxyethylene sorbitan fatty acid esters (e.g. ), polyoxyethylene fatty acid glycerides (e.g. ), polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, glycerol fatty acid esters, poloxamers (e.g., ),

[0536] Buffers, acids and bases (e.g. phosphates, carbonates, citric acid, acetic acid, hydrochloric acid, sodium hydroxide solution, ammonium carbonate, tromethamine, triethanolamine),

[0537] Isotonic agents (e.g. glucose, sodium chloride),

[0538] Adsorbents (e.g. highly dispersed silica),

[0539] Viscosifiers, gel formers, thickeners and / or binders (e.g., polyvinylpyrrolidone, methylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, starch, carbomer, polyacrylic acid (e.g., ); alginate, gelatin),

[0540] Disintegrants (e.g. modified starch, sodium carboxymethylcellulose, sodium starch glycolate (e.g. ), cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose (e.g., )),

[0541] Flow regulators, lubricants, glidants and release agents (e.g. magnesium stearate, stearic acid, talc, highly dispersed silicon dioxide (e.g. )),

[0542] Coating materials (e.g. sugar, shellac) and film formers for membranes or diffusion membranes that dissolve rapidly or in a modified manner (e.g. polyvinylpyrrolidone (e.g. ), polyvinyl alcohol, hydroxypropyl methylcellulose, hydroxypropyl cellulose, ethyl cellulose, hydroxypropyl methylcellulose phthalate, cellulose acetate, cellulose acetate phthalate, polyacrylates, polymethacrylates, e.g. )),

[0543] capsule materials (e.g., gelatin, hydroxypropyl methylcellulose),

[0544] Synthetic polymers (e.g. polylactide, polyglycolide, polyacrylates, polymethacrylates (e.g. ), polyvinylpyrrolidone (e.g. ), polyvinyl alcohol, polyvinyl acetate, polyethylene oxide, polyethylene glycol and copolymers and block copolymers thereof,

[0545] Plasticizers (e.g. polyethylene glycol, propylene glycol, glycerol, triacetin, triacetyl citrate, dibutyl phthalate),

[0546] Penetration enhancers,

[0547] Stabilizers (e.g. antioxidants, such as ascorbic acid, ascorbyl palmitate, sodium ascorbate, butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate),

[0548] Preservatives (e.g. parabens, sorbic acid, thiomersal, benzalkonium chloride, chlorhexidine acetate, sodium benzoate),

[0549] Colorants (e.g. inorganic pigments, such as iron oxide, titanium dioxide),

[0550] • Flavoring, sweetening, flavoring and / or odor-masking agents.

[0551] The present invention also relates to a pharmaceutical composition 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.

[0552] In particular, the present invention relates to a pharmaceutical composition 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, usually together with one or more pharmaceutically suitable excipients, and their use according to the invention.

[0553] The pharmaceutical composition of the present invention may comprise at least one further active ingredient, eg a further active ingredient which is preferably active in the prevention and / or treatment of a disorder or disease as defined herein.

[0554] At least one peptide, derivative or analogue as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex 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 orally. Suitable formulations for the respective routes of administration are well known to the skilled person and include, but are not limited to, pills, tablets, enteric-coated tablets, films, laminates, sustained-release or extended-release formulations for oral administration, plasters, topical sustained-release formulations, dragees, pessaries, gels, ointments, syrups, granules, suppositories, emulsions, dispersions, microcapsules, microformulations, nanoformulations, liposomal formulations, capsules, enteric-coated capsules, powders, inhalation powders, microcrystalline formulations, inhalation sprays, powders, drops, nasal drops, nasal sprays, aerosols, ampoules, solutions, juices, suspensions, infusion solutions or injection solutions, etc.

[0555] According to another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound containing a peptide (which can be isolated and / or purified) or a derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, in combination with one or more inert, non-toxic, pharmaceutically suitable excipients, wherein the peptide comprises, essentially consists of or consists of formula (I) or formula (II), and the pharmaceutical composition is used to prevent and / or treat cardiovascular and cardiopulmonary diseases, shock, inflammatory diseases, cardiovascular, pulmonary, cerebral and renal sequelae of sepsis, ischemia and / or reperfusion-related injury, acute kidney injury, transplant protection and delayed graft function, diseases of the blood and hematopoietic organs and the immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, respiratory system diseases, digestive system diseases or genitourinary system diseases, as well as sequelae of burns and sequelae of injury.

[0556] According to another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound comprising a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, in combination with one or more other active ingredients, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II), and wherein the other active ingredients are selected from the group consisting of: phosphodiesterase inhibitors, guanylate cyclase stimulators or activators, IP receptor agonists, mineralocorticoid receptor antagonists, diuretics, PPAR-γ agonists, PPAR-δ agonists, corticosteroids, active ingredients that reduce organ damage under oxidative stress, inhibit induced cell death. and apoptosis 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 coagulation factor XI inhibitors for the prevention and / or treatment of cardiovascular and cardiopulmonary diseases, shock, inflammatory diseases, cardiovascular, pulmonary, cerebral and renal sequelae of sepsis, ischemia and / or reperfusion-related injury, acute kidney injury, transplant protection and delayed graft function, diseases of the blood and hematopoietic organs and immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, respiratory system diseases, digestive system diseases or genitourinary system diseases and sequelae of burns and injuries.

[0557] According to another embodiment, the present invention provides a method for treating and / or preventing the following diseases in humans and animals: cardiovascular and cardiopulmonary diseases, shock, inflammatory diseases, cardiovascular, pulmonary, cerebral and renal sequelae of sepsis, damage associated with ischemia and / or reperfusion, acute kidney injury, transplant protection and delayed graft function, diseases of the blood and hematopoietic organs and the immune system, sequelae of diabetes, inflammatory diseases of the nervous system, eye diseases, skin diseases, respiratory system diseases, digestive system diseases or genitourinary system diseases, sequelae of burns and injuries by administering an effective amount of a pharmaceutical composition comprising at least one peptide or derivative that can be isolated and / or purified , prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt, the peptide comprising, consisting essentially of or consisting of formula (I) or formula (II), or the pharmaceutical composition comprising at least one compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, in combination with one or more inert, non-toxic, pharmaceutically suitable excipients and / or one or more other active ingredients.

[0558] The appropriate dosage of the MASP inhibitory peptides of the present invention can be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dosage level for any particular subject will depend on a variety of factors, including: a) the condition being treated and the severity of the condition; 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 concurrently with the MASP inhibitory peptide used, and similar factors well known in the medical art.

[0559] In certain embodiments, the total daily dose of the MASP inhibitory peptides of the invention administered to a subject or patient in a single or divided dose can be, for example, 0.0001 to 300 mg / kg body weight per day, or 1 to 300 mg / kg body weight per day, or about 0.0001 to about 100 mg / kg body weight per day, for example, about 0.0005 to about 50 mg / kg body weight per day, for example, about 0.001 to about 10 mg / kg body weight per day, for example, about 0.01 to about 1 mg / kg body weight per day, administered in one or more doses, for example, one to three doses. Generally, the MASP inhibitory peptides of the invention can be administered continuously (e.g., by intravenous administration or by another continuous administration method), or can be administered to the subject intermittently, generally at fixed time intervals, depending on the desired dose and the pharmaceutical composition selected by the skilled practitioner for the particular subject. Fixed dosing intervals include, for example, once a day, twice a day, every two, three, four, five or six days, once or twice a week, once or twice a month, etc.

[0560] The present invention also encompasses the use of a MASP inhibitory peptide as described herein for the preparation of a medicament, in particular for the preparation of a medicament for the prevention and / or treatment of a condition or disease as defined herein.

[0561] The present invention also includes a method for preparing the peptides, derivatives or analogs thereof, or pharmaceutically acceptable salts or solvates or complexes thereof (each as described herein), which may include the steps shown in the examples of the present invention.

[0562] Generally, the MASP inhibitory peptides of the present invention can be prepared synthetically or semi-recombinantly.

[0563] According to another embodiment, the present invention provides a method for preparing a compound containing a peptide or a derivative, prodrug, analog or pharmaceutically acceptable salt or solvate thereof that can be isolated and / or purified by using solid phase polypeptide synthesis, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II).

[0564] According to another embodiment, the present invention provides a method for preparing a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II), the method comprising the steps of:

[0565] 1. Use 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,

[0566] 2. Load the C-terminal amino acid of the sequence onto the resin,

[0567] 3. Remove the fmoc protection with 15-25% piperidine solution in DMF or NMP,

[0568] 4. Couple the next amino acid in the sequence with a coupling reagent (e.g., HBTU, HATU, or DICC / Oxyma) using a stoichiometry of 3-8 equivalents,

[0569] 5. Repeat steps 3 and 4 until the sequence is complete,

[0570] 6. Cleavage of the peptide from the solid support using a cleavage mixture containing TFA and a thiol scavenger,

[0571] 7. Cyclize the two cysteines in the sequence under oxidative conditions (air or I2),

[0572] 8. Purify the cleaved peptide using reverse phase HPLC.

[0573] According to another embodiment, the present invention provides a method for preparing a compound containing a peptide or derivative, prodrug, analog, pharmaceutically acceptable salt, solvate or solvate of a salt that can be isolated and / or purified, wherein the peptide comprises, consists essentially of or consists of formula (I) or formula (II), the method comprising the steps of:

[0574] 1. Use 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,

[0575] 2. Load the C-terminal amino acid of the sequence onto the resin,

[0576] 3. Remove the fmoc protection with 15-25% piperidine solution in DMF or NMP,

[0577] 4. Couple the next amino acid in the sequence with a coupling reagent (e.g., HBTU, HATU, or DIC / Oxyma) using a stoichiometry of 3-8 equivalents,

[0578] 5. Repeat steps 3 and 4 until the sequence is complete,

[0579] 6. Cleavage of the peptide from the solid support using a cleavage mixture containing TFA and a thiol scavenger,

[0580] 7. Cyclize the two cysteines in the sequence under oxidative conditions (air or I2),

[0581] 8. Purify the cleaved peptide using reverse phase HPLC,

[0582] 9. Converted to HCl salt.

[0583] At least one peptide, derivative or analog as defined herein, or a pharmaceutically acceptable salt or solvate thereof, or a complex as defined herein, can also be used as a biochemical reagent in a biochemical assay for measuring responsiveness to a MASP inhibitory agent (e.g., in a diagnostic assay) or in any biochemical assay based on binding of a MASP inhibitory agent.

[0584] The present invention also includes polynucleotides comprising sequences encoding the MASP inhibitory peptides of the present invention and vectors comprising polynucleotides comprising sequences encoding the MASP inhibitory peptides of the present invention.

[0585] The present invention is further illustrated by the following examples, which relate to certain specific embodiments of the present invention. Unless otherwise stated, examples are performed using standard techniques well known to those skilled in the art within the conventional range. The following examples are intended for illustrative purposes only and are not intended to fully define 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. Specific implementation plan

[0586] List of abbreviations used in the experimental section:

[0587] Angstrom

[0588] aq. water-containing, aqueous solution

[0589] bar pressure unit

[0590] BPR Back Pressure Regulator

[0591] conc concentrated

[0592] d Doublet (NMR)

[0593] dd Doublet (NMR)

[0594] DCM dichloromethane

[0595] DEA Diethylamine

[0596] DiPEA N,N,-diisopropylethylamine (Hünig's base)

[0597] DMAP 4-dimethylaminopyridine

[0598] DMF N,N-dimethylformamide

[0599] DMSO dimethyl sulfoxide

[0600] dt double triplet (NMR)

[0601] EA Ethyl acetate

[0602] ee enantiomeric excess

[0603] ent enantiomer

[0604] eq equivalent

[0605] equiv equivalent (ion chromatography)

[0606] ESI Electrospray Ionization (Mass Spectrometry)

[0607] Fmoc-OSu 1-{[(9H-fluoren-9-ylmethoxy)carbonyl]oxy}pyrrolidine-2,5-dione

[0608] GC-MS gas chromatography coupled with mass spectrometry

[0609] h hour

[0610] HATU O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0611] HPLC High-pressure liquid chromatography

[0612] IC ion chromatography

[0613] ID inner diameter

[0614] L liter

[0615] LC-MS liquid chromatography coupled with mass spectrometry

[0616] LiHMDS Lithium bis(trimethylsilyl)amide

[0617] lit. Literature

[0618] m multiplet (NMR)

[0619] MAL II matrix-assisted laser desorption / ionization (MS)

[0620] Me methyl

[0621] min

[0622] mm millimeter

[0623] μm micrometer (micron)

[0624] M Moore

[0625] MPLC Medium Pressure Liquid Chromatography

[0626] MS

[0627] MTBE tert-butyl methyl ether

[0628] MTP microtiter plates

[0629] m / z mass-to-charge ratio (mass spectrometry)

[0630] nm nanometer

[0631] NMP N-Methyl-2-pyrrolidone

[0632] NMR nuclear magnetic resonance spectroscopy

[0633] PBS Phosphate-buffered saline

[0634] PE petroleum ether

[0635] PEG polyethylene glycol

[0636] pos positive

[0637] ppm parts per million

[0638] Pr Propyl

[0639] Psi pounds per square inch (pressure)

[0640] q / quart quartet (NMR)

[0641] qd four doublets (NMR)

[0642] quint quintet (NMR)

[0643] rac racemic

[0644] Rf retention factor (TLC)

[0645] RP Reversed Phase (for liquid chromatography)

[0646] Rt retention time (chromatography)

[0647] s singlet (NMR)

[0648] s seconds (time)

[0649] SPPS Solid Phase Peptide Synthesis

[0650] t triplet (NMR)

[0651] TBTU O(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate

[0652] tBU tert-butyl

[0653] TEA triethylamine

[0654] THF Tetrahydrofuran

[0655] TLC thin layer chromatography

[0656] UPLC Ultra-Performance Liquid Chromatography

[0657] UV

[0658] Other abbreviations can be found in Tables 2 and 3.

[0659] LC-MS analysis

[0660] Method 1

[0661] Equipment type MS: ThermoFisherScientific LTQ-Orbitrap-XL; Equipment type HPLC: Agilent 1200SL; Column: Agilent, POROSHELL 120, 3x150 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.

[0662] Method 2

[0663] Equipment type MS: ThermoFisherScientific LTQ-Orbitrap-XL; Equipment type HPLC: Agilent 1200SL; Column: Agilent, POROSHELL 120; 3x150mm, SB-C182.7μm; Eluent A: 1L water + 0.1% trifluoroacetic acid; Eluent B: 1L acetonitrile + 0.1% trifluoroacetic acid; Gradient: 0.0min 5% B→0.3min 5% B→7.0min 98% B→10min 98% B; Oven: 40℃; Flow rate: 0.75mL / min; UV-detection: 210nm.

[0664] Method 3

[0665] Equipment type MS: Waters TOF instrument; Equipment type UPLC: Waters Acquity I-CLASS; Column: YMC, TRIART C18, 75x1mm, 3.0μm x 12nm; Eluent A: 1L water + 0.01% formic acid; Eluent B: 1L acetonitrile + 0.01% formic acid; Gradient: 0.0min 1% B→2.0min 1% B→8.0min 95% B→10.0min 95% B; Oven: 50°C; Flow rate: 0.63mL / min; UV-detection: 210nm.

[0666] Method 4

[0667] Equipment type MS: Waters TOF instrument; Equipment type UPLC: Waters Acquity I-CLASS; Column: YMC, TRIART C18, 75x1mm, 3.0μm x 12nm; Eluent A: 1L water + 0.01% formic acid; Eluent B: 1L acetonitrile + 0.01% formic acid; Gradient: 0.0min 1% B→1.0min 1% B→15.0min 50% B→18.0min 95% B; Oven: 50°C; Flow rate: 0.63mL / min; UV-detection: 210nm.

[0668] Method 5

[0669] Equipment type MS: Waters TOF instrument; Equipment type UPLC: Waters Acquity I-CLASS; Column: Waters, HSST3, 2.1x50 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.

[0670] Method 6

[0671] Equipment type MS: Waters Synapt G2S; equipment type UPLC: Waters Acquity I-CLASS; column: Waters, BEH300, 2.1x150mm, C181.7μm; eluent A: 1L water + 0.01% formic acid; eluent B: 1L acetonitrile + 0.01% formic acid; gradient: 0.0min 2%B→1.5min 2%B→8.5min 95%B→10.0min 95%B; oven: 50℃; flow rate: 0.50mL / min; UV-detection: 220nm.

[0672] Method 7

[0673] Instrument type MS: Agilent 6410 Triple Quad; Instrument type HPLC: Agilent 1200; Column: Gemini-NX C18 5μm 150x4.6mm; eluent A: 0.1% TFA in water; eluent B: 0.1% TFA in acetonitrile; gradient: 0.0min 20% B→20min 50% B→20.1min 90% B→23min 90% B; oven temperature: 50°C; flow rate: 1.0mL / min; UV-detection: 220nm.

[0674] Method 8

[0675] Instrument type MS: Agilent 6410Triple Quad; Instrument type HPLC: Agilent 1200; Column: Discovery BIO Wide Pore C18 5μm x4.6 mm; eluent A: 0.1% TFA in water; eluent B: 0.1% TFA in acetonitrile; 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.

[0676] Method 9

[0677] Instrument: Waters ACQUITY SQD UPLC system; column: Waters Acquity UPLC HSS T3 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 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.

[0678] Method 10

[0679] Instrument MS: Thermo Scientific FT-MS; Instrument UHPLC: Thermo Scientific UltiMate3000; Column: Waters, HSST3, 2.1x75mm, C181.8μm; Eluent A: 1L water + 0.01% formic acid; Eluent B: 1L acetonitrile + 0.01% formic acid; Gradient: 0.0min 10% B→2.5min 95% B→3.5min 95% B; Oven: 50°C; Flow rate: 0.90mL / min; UV-detection: 210nm / optimal integration path 210-300nm.

[0680] Method 11

[0681] MS instrument: Agilent MS Quad6150; HPLC: Agilent 1290; column: Waters Acquity UPLCHSS T3 1.8μm 50x2.1mm; eluent A: 1L water + 0.25mL formic acid, eluent B: 1L acetonitrile + 0.25mL formic acid; gradient: 0.0min 90% A→0.3min 90% A→1.7min 5% A→3.0min 5% A; oven: 50°C; flow rate: 1.20mL / min; UV-detection: 205-305nm.

[0682] Method 12

[0683] Instrument: Waters ACQUITY SQD UPLC system; column: Waters Acquity UPLC HSS T3 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.

[0684] Method 13

[0685] Instrument: Waters Single Quad MS system; Instrument: Waters UPLC Acquity; Column: Waters BEHC18 1.7 μm 50x2.1 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.

[0686] Method 14

[0687] System MS: Waters ToF instrument; UPLC system: Waters Acquity I-CLASS; column: Waters Acquity UPLC Peptide BEH C18 1.7 μm 150x2.1 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 nmol / s

[0688] Method 15

[0689] MS instrument type: Agilent G6110A; HPLC type: Agilent 1200 Series LC; UV DAD; column: Chromolith RP-18e 25x2.0 mm; mobile phase A: 0.0375% TFA in water (v / v), mobile phase B: 0.01875% TFA in acetonitrile (v / v); 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.

[0690] MALDI method

[0691] Accurate mass measurements of selected peptides were performed using Matrix Assisted Laser Desorption / Ionization (MALDI) mass spectrometry on a Brukerautoflex 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 a matrix. Solutions of 0.1 to 1.0 mg of the sample peptide in 1.0 mL of acetonitrile-water (50 / 50 or 30 / 70) and a stock solution of the matrix (10 mg / mL) in 50% acetonitrile in water containing 0.05% trifluoroacetic acid were prepared. 1.0 uL of each solution was placed on a MALDI target plate and allowed to dry. The sample was then ready for analysis. Recommended sample preparation for MALDI target plates can be found in the literature provided by Bruker.

[0692] Ion chromatography

[0693] Method: IC-quantification

[0694] External standards were used for quantitative measurement of cations and anions. Instrument: Thermo Scientific ICS 5000+. Capillary IC columns: IonPac AS11-HC and IonPac CS16. Eluent: Gradient eluent [H]+[OH]-. Detection: Conductivity. Possible common anions: acetate, bromide, citrate, chloride, fluoride, formate, lactate, methanesulfonate, phosphate, sulfate, tartrate, trifluoroacetate. Possible common cations: ammonium, barium, calcium, potassium, lithium, sodium, magnesium, choline.

[0695] Gas chromatography-mass spectrometry

[0696] GC-MS method 1

[0697] Instrument: Thermo Scientific DSQII, Thermo Scientific Trace GC Ultra; Column: Restek RTX-35MS, 15 m x 200 μm x 0.33 μm; Helium constant flow: 1.20 mL / min; Oven: 60°C; Inlet: 220°C; Gradient: 60°C, 30°C / min→300°C (hold for 3.33 min).

[0698] NMR

[0699] The selected compounds 1 H-NMR data were 1H-NMR peak lists are presented in the form of a peak list. For each singlet, the δ value is given in ppm, followed by the signal intensity reported in parentheses. Delta value-signal intensity pairs from different peaks are separated by commas. Thus, peak lists are described using the general form: δ1 (intensity 1), δ2 (intensity 2), ..., δi (intensity i), ..., δn (intensity n).

[0700] The sharp signal intensity is related to the height of the signal in the printed NMR spectrum (in centimeters). This data can be correlated to the actual ratio of the signal intensities when compared to other signals. In the case of broad signals, more than one peak or signal center is shown, along with their relative intensities, compared to the strongest signal displayed in the spectrum. 1 The H-NMR peak list resembles a typical 1 H-NMR readings therefore typically contain all the peaks listed in a typical NMR interpretation. In addition, similar to a typical 1 In the H-NMR printout, the peak list can show solvent signals, signals originating from stereoisomers of a particular target compound, impurity peaks, 13C satellite peaks and / or spinning sidebands. The peaks of stereoisomers and / or peaks of impurities are usually shown with lower intensity (e.g. purity > 90%) compared to the peaks of the target compound. Such stereoisomers and / or impurities may be typical for a particular preparation process, so their peaks may help to identify the reproducibility of the preparation process based on a "by-product fingerprint". An expert who has calculated the peaks of the target compound by known methods (MestReC, ACD simulation or using empirically assessed expectation values) can separate the peaks of the target compound as needed, optionally using other intensity filters. This operation is similar to a typical 1 Peak picking in H-NMR interpretation. Detailed instructions for reporting NMR data in peak list format can be found in the publication "Citation of NMR Peaklist Data within Patent Applications" (cf. http: / / www.researchdisclosure.com / searching-disclosures, Research Disclosure Database Number 605005, 2014, August 1, 2014). In the peak picking procedure, as described in Research Disclosure Database Number 605005, the parameter "Minimum Height" can be adjusted between 1% and 4%. However, depending on the chemical structure and / or the concentration of the compound being tested, it may be reasonable to set the parameter "Minimum Height" to <1%.

[0701] Preparation Example

[0702] General method of synthesis

[0703] Solid phase peptide synthesis (SPPS) was performed using an automated peptide synthesizer or manually. Peptide synthesis was typically performed on a scale of 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).

[0704] Fmoc-protected amino acids (or intermediate amino acids for the preparation of Fmoc amino acids) were purchased from Novabiochem, Bachem, Iris Biotech, Sigma-Aldrich, Alfa, Enamine, Amatek, Anichem, ACBR, ABCLaboratory, AP Bioscience, Combiblocks, ArZa Bioscience, Ark Pharm, Acroteinchem, Apollo Scientific, Biofine, Broadpharm, VWR or Gyros Protein Technologies (fluorenylmethoxycarbonyl=Fmoc), GL Biochem (Shanghai) Ltd, Chengdu Aminotp Pharmaceutical Technology Ltd, Suzhou Highfine Biotech Co., Ltd, WuXi AppTec, or purchased through other Chinese suppliers. Some special fmoc-protected amino acids were synthesized in-house, and these synthetic methods are described herein. Some in-house synthesized fmoc amino acids are also commercially available. In some cases, Fmoc-protected amino acids are not commercially available, but Boc-protected (tert-butyloxycarbonyl=Boc) non-natural amino acids are commercially available, and the Fmoc-protected amino acids are prepared from the Boc-protected amino acids by deprotection and reprotection using methods commonly used in the art. In most cases, the CAS numbers of the commercially available non-natural amino acids used to synthesize the peptides of the present invention are included in Table 5. In the case of purchasing racemic amino acids (Fmoc or Boc), those skilled in the art will recognize that enantiomers can be separated using chiral chromatography, and in fact this is sometimes done to select enantiomerically pure amino acids prior to peptide synthesis.

[0705] The following unnatural amino acids have been used to prepare the peptides of the present invention. Fmoc- or Boc-protected amino acids were either obtained from commercial sources (CAS numbers available) or synthesized in-house using methods described herein. Table 5 shows the CAS numbers of the chemical groups / amino acids used for peptide synthesis (right column) and the corresponding chemical groups / amino acids present in the peptides (left column).

[0706] Table 5: Availability of unnatural amino acids and chemical groups of the present invention

[0707]

[0708]

[0709]

[0710]

[0711]

[0712]

[0713]

[0714]

[0715]

[0716]

[0717] Solid phase resins were purchased from Novabiochem, Bachem, IrisBiotech, PcasBiomatrix, GLBiochem (Shanghai) Ltd, CEM or Protein Technologies. Resin loading was 0.3-1.0 mmol / g. Peptides were synthesized on 2-chlorotrityl resin, Wang resin or Rink amide 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 using 8 equivalents of Fmoc-amino acid with 8 equivalents of DIC (diisopropylcarbodiimide) (0.5 M in DMF) and 8 equivalents of Oxyma (ethyl cyanohydroxyiminoacetate) (0.5 M in DMF) on an automated synthesizer (Symphony X). When using the Symphony X, amino acid coupling was performed at room temperature under a nitrogen atmosphere. When using expensive or self-prepared fmoc-amino acids, 3 equivalents of Fmoc-amino acids are manually coupled 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 is performed twice (double coupling). Alternatively, 3 equivalents of Fmoc-amino acids, 2.85 equivalents of HBTU ((2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, hexafluorophosphate benzotriazole tetramethyluronium (Hexafluorophosphate Benzotriazole TetramethylUronium) (0.5 M in DMF)) and 6 equivalents of DIPEA (0.5 M in DMF) are manually prepared by SPPS. The coupling reaction is monitored using the ninhydrin test.

[0718] The peptides were completely 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% water. For peptides containing methionine, the peptides were treated with 1.6% EDT and 1.2% trimethylsilyl bromide in TFA for 2 hours at room temperature to reduce the oxidized methionine.

[0719] Disulfide cyclization:

[0720] Disulfide bonds are formed by shaking a 0.5 mg / mL peptide overnight in 0.1 M ammonium bicarbonate buffer (pH 7.83). The solution is then lyophilized. Alternatively, disulfide bonds are formed by shaking the peptide overnight in an acetonitrile / water (typically 3:7) mixture adjusted to pH 9.0 with solid ammonium bicarbonate buffer at a concentration of 1-3 mg / mL. Alternatively, disulfide bonds are prepared by oxidizing a 1-1.3 mg / mL peptide with iodine (I2) (0.1 M in methanol) in acetonitrile / water (1:1) at 20°C for 2 minutes, followed by treatment with sodium thiosulfate (0.1 M in water) and lyophilization.

[0721] Optional acetylation:

[0722] N-terminal acetylation was performed by shaking the suspension on an orbital shaker for 1 hour using 10 equivalents of acetic anhydride in DMF (2 mL) and 2.5 equivalents of DIPEA. The solvent was removed and the resin was washed with DMF (5x) and DCM (5x). The process was then repeated again. Alternatively, 10 mL of the end-capping solution consisting of acetic anhydride / N-methylmorpholine (NMM) / DMF (10: 5: 85) was used to shake the suspension on an orbital shaker for 30 minutes.

[0723] Cleavage peptide:

[0724] Prepare a cleavage mixture containing TFA / EDT / thioanisol (90:3:7). Add the cleavage mixture (2 mL) to the resin containing the peptide, and shake the suspension on an orbital shaker for 2.5 hours. Add cold ether (-20°C) to precipitate the peptide. The resulting solution is centrifuged under nitrogen (Sigma 2-16KL), and the resulting solid, after decantation, is washed three more times with cold ether by centrifugation and decantation. The resulting solid is purified by preparative HPLC.

[0725] Alternatively, a cleavage mixture containing TFA / EDT / TIS / H2O (92.5: 2.5: 2.5: 2.5) was prepared. The cleavage mixture (6 mL (0.3 mmol scale)) was added to the resin containing the peptide, and the suspension was shaken on an orbital shaker for 2.5 hours. Cold tert-butyl methyl ether (-20°C) was added to precipitate the peptide. The resulting solution was centrifuged at 3000 rpm for 3 minutes, and the solid obtained after decantation was washed more than 3 times (20 mL x 3) with cold tert-butyl methyl ether by centrifugation and decantation. The crude peptide was vacuum dried for 2 hours and then purified by preparative HPLC.

[0726] Preparative HPLC:

[0727] Purification was performed using an Agilent 1260 Prep reverse phase HPLC or a Knauer AZURA Prep reverse phase HPLC. Column selection was based on the results of the column screening. The peptide was dissolved in 10-30% ACN / water (typically the starting point of the gradient). Both the water and acetonitrile contained 0.1% TFA. A flow rate of 20 mL / min from 10-30% ACN / water to 85-90% ACN / water was typically used. A Chromolith Speedrod column was used with a 5-95% ACN / water gradient over 8 minutes by HPLC (Agilent 1260 Infinity) and fractions were analyzed by one or more of the following LC-MS methods: Method 1, Method 2, Method 3, Method 4, Method 5, Method 6.

[0728] Alternatively, purification was performed using a Gilson GX-281 Prep reverse phase HPLC. The column was selected based on the results of the column screening. The peptide was dissolved in 10-30% acetonitrile / water (usually the starting point of the gradient). The water contained 0.075% TFA. A Luna column (25x200mm, C18 10μm, ) or Gemini column (30x150mm, C185μm, Preparative HPLC conditions: flow rate 20 mL / min, 10-30% ACN / water to 85-90% ACN / water, wavelength 214 / 254 nm, oven temperature 30°C. Fractions were analyzed by HPLC (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.

[0729] Disulfide mimetics (wherein -SS-disulfide bonds are replaced by -CH2-S-, -S-CH2-, -CH2-CH2-, -S-(CH2)2-, -(CH2)2-S-, or -CH2-S-CH2-) can be prepared according to the procedures described in the following references in combination with the methods described herein: (1) Hong-Kui Cui, Ye Guo, Yao He, Feng-Liang Wang, Ha0-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 Xu, Donald Bierer, Jun Wang, Jing Shi, Yi-Ming Li Tetrahedron Letters 2017, 58, 3970-3973; (5) Tao Wang, Jian Fan, Xiao-Xu Chen, Rui Zhao, Yang Xu, Donald Bierer, Lei Liu, Yi-Ming Li, Jing Shi, Ge-Min Fang Org. Lett. 2018, 20, 6074-6078; (6) Jan-Patrick Fischer, Ria 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) CMBK Kourra and N. Cramer Chem. Sci., 2016, 7, 7007-7012.

[0730] Unless otherwise stated, all peptides of the present invention are TFA salts.

[0731] General method for automated SPPS of Masp peptides (Method A)

[0732] The synthesis of ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IPD-NH2 (Example 165) is representative.

[0733] The peptide was synthesized using standard Fmoc chemistry.

[0734] Automated SPPS was performed on a Symphony X peptide synthesizer (Protein Technologies). Chem-MatrixRink amide resin was typically used at a 0.1 mmol scale (loading of 0.5 mmol / g). The resin was placed in a reaction vessel and placed on the instrument. The following solutions were prepared and used during the synthesis:

[0735] 1) Fmoc amino acid: 0.2M (8eq)

[0736] 2) Activator 1: 0.5M DIC in DMF (7.5eq)

[0737] 3) Activator 2: 0.5M Oxyma DMF solution (7.5eq)

[0738] 4) Fmoc deprotection: 20% piperidine in DMF solution

[0739] 5) Acetic anhydride (10 equivalents) in 2 mL DMF

[0740] A double coupling is typically performed for each amino acid. For expensive non-natural Fmoc or Boc amino acids, in-house synthesized Fmoc amino acids, or N-methylated amino acids, the sequence is interrupted and the amino acid is coupled manually (double coupling, but typically with less reagent (3-5 equivalents)). After this coupling is complete, the synthesis is typically continued on the synthesizer. If an N-methyl amino acid is added to the sequence, the next amino acid is typically coupled manually as well. All steps are performed at room temperature under nitrogen. Fmoc-Pen and Fmoc-Oic are typically coupled using automated SPPS. Fmoc(N-Me)G is typically coupled manually. Ahx is always coupled manually.

[0741] The resin was swollen and washed with DMF (3 x 3 mL, 10 min). If the resin contained Fmoc, it was removed with 20% piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 sec).

[0742] Coupling:

[0743] Add Fmoc-Asp (t-Bu) (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3mL, 30 seconds). Repeat the coupling step. Add Fmoc-Asp (Ot-Bu) (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3mL, 30 seconds).

[0744] Fmoc cleavage:

[0745] 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 sec).

[0746] Coupling:

[0747] Add Fmoc-Pro (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours. Drain solution and wash with DMF (1x3mL, 30 seconds). Repeat coupling step. Add Fmoc-Pro (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours. Drain solution and wash with DMF (6x3mL, 30 seconds).

[0748] Fmoc cleavage:

[0749] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0750] Coupling:

[0751] Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0752] Fmoc cleavage:

[0753] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0754] Coupling:

[0755] Add Fmoc-Pen (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Pen (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0756] Fmoc cleavage:

[0757] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0758] Coupling:

[0759] Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0760] Fmoc cleavage:

[0761] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0762] Coupling:

[0763] Add Fmoc-Oic (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Oic (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0764] Fmoc cleavage:

[0765] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0766] Coupling:

[0767] Add Fmoc-Pro (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours. Drain solution and wash with DMF (1x3mL, 30 seconds). Repeat coupling step. Add Fmoc-Pro (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours. Drain solution and wash with DMF (6x3mL, 30 seconds).

[0768] Fmoc cleavage:

[0769] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0770] Coupling:

[0771] Add Fmoc-Leu (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and allow the coupling to proceed under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Leu (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and allow the coupling to proceed under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0772] Fmoc cleavage:

[0773] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0774] Coupling:

[0775] Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0776] Fmoc cleavage:

[0777] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0778] Coupling:

[0779] Add Fmoc-Arg (Pbf) (4.0mL). Add activator 12 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3mL, 30 seconds). Repeat the coupling step. Add Fmoc-Arg (Pbf) (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3mL, 30 seconds).

[0780] Fmoc cleavage:

[0781] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0782] Coupling:

[0783] Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0784] Fmoc cleavage:

[0785] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0786] Coupling:

[0787] Add Fmoc-Cys (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Cys (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0788] Fmoc cleavage:

[0789] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0790] Coupling:

[0791] Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0792] Fmoc:

[0793] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0794] Manual coupling during automated SPPS:

[0795] Fmoc-(N-Me) G (0.2M DMF solution, 5 equivalents) is added to the resin. Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL), and couple for 2 hours under vibration (thermomixer (Thermomixer), room temperature). Filter the solution and wash with DMF (1x3mL, 30 seconds). Repeat the coupling step. Add Fmoc-(N-Me) G (0.2M DMF solution, 5 equivalents). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL), and couple for 2 hours under vibration (thermomixer, room temperature). Filter the solution and wash with DMF (6x3mL, 30 seconds).

[0796] Fmoc cleavage:

[0797] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0798] If additional amino acids were present in the sequence, they were coupled using the procedure described above.

[0799] Lysis test:

[0800] When performing manual coupling, cleavage is usually performed test To monitor the reaction. test The cleavage mixture was TFA / EDT / thioanisole (90:3:7); shaken at 750 rpm on a thermomixer at room temperature for 1.5 hours. Analysis was performed by LC-MS using one of the methods described above.

[0801] Full cleavage:

[0802] The resin containing the peptide was placed in a syringe and 3.0 mL of lysis 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 collected by filtration and the peptide was precipitated by adding cold ether (-20°C). Under a nitrogen atmosphere, the solution was centrifuged (3000 rpm) in a Falcon tube (60 mL). The ether was decanted and the solid residue was repeatedly washed with cold ether (5 x 10 mL). The solid residue was then dried.

[0803] Disulfide cyclization:

[0804] 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 overnight in a round-bottom flask open to air on an orbital shaker. The solution was then lyophilized to obtain a white powder.

[0805] Column screening for HPLC purification:

[0806] The peptides were dissolved in 5% CH3CN and 95% water. Column screening was performed for each peptide to determine the preparative HPLC method for purification. The following analytical columns were screened.

[0807] Two methods can be used for column screening:

[0808] 1)5-60%ACN_8Min_1mL / min_25℃

[0809] 2)30-85%ACN_8Min_1mL / min_25℃

[0810] Available columns (50mm x ID 4, 6mm) (can also be used as preparative columns):

[0811] 1) Aeris C18 (Phenomenex)

[0812] 2) X-Bridge C18 (Waters)

[0813] 3)Kinetex C18(Coreshell Material)(Phenomenex)

[0814] 4) YMC Triart C18 (elutable with 100% water)

[0815] 5)Kinetix Biphenyl(Phenomenex)

[0816] 6) X-Select C18 (positive charge)

[0817] 7)Jupiter Proteo C18 (Phenomenex)

[0818] 8) Luna C18 (Phenomenex)

[0819] For the peptides of the present invention, one of the following five preparative columns was used:

[0820] 1) Column: Phenomenex, Aeris Peptide 5μ XB-C18, AXIA Packed, 21, 2x250mm+Cartridge 5μ

[0821] 2) Column: Phenomenex, Kinetex C18 5μ 21, 5x250mm+Cartridge 5μ

[0822] 3) Column: Phenomenex, Kinetex 5μ Biphenyl 100A, AXIA Packed, 21, 2x250mm+Cartridge 5μ

[0823] 4) Column: YMC Actus triart Prep.C1812nm, S-10μm 250x20mm+Cartridge 3μm (10x4mm)

[0824] 5) Column: Waters, Xbridge Prep.C18 5μ OBD 19x250mm+Cartridge 10μ

[0825] After selecting a column, use one of the following methods:

[0826] 1) Method: Gradient 5-60% ACN aqueous solution (0, 10% TFA)

[0827] 2) Method: Gradient 30-85% ACN aqueous solution (0, 10% TFA)

[0828] 3) Focused gradient based on column screening results.

[0829] Flow rate 20mL / min

[0830] The combined fractions were analyzed by HPLC (5-95 in 8 min, Chromolith SpeedROD & YMC C18 5-95 in 18 min) and using one of the LC-MS methods described above.

[0831] For Example 165, the crude peptide was dissolved in CH3CN / water (1:1) and purified on a Phenomenex Kinetex column C18 5μ, 21.5x250mm+5μCartridge, flow rate: 20mL / min, method: 5-60% aqueous ACN (each containing 0.10% TFA) over 48 minutes. The combined fractions were lyophilized to provide 39 mg (95% purity) of Example 165.

[0832] Table 6: Description of materials and conditions used

[0833]

[0834] General method for automated SPPS of MasD peptides (Method B)

[0835] 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.

[0836] The peptide was synthesized using standard Fmoc chemistry.

[0837] If possible, use Wang resin preloaded with the first amino acid (e.g., preloaded with Fmoc-Asp(t-Bu)-OH, Fmoc-Pro-OH, etc.), typically with a loading of 0.55-0.7 mmol / g. When the first amino acid is not available, add it manually.

[0838] When the first amino acid was not available as preloaded resin, it was added manually and synthesized using chlorotrityl resin.

[0839] Loading of the first amino acid on 2-chlorotrityl resin:

[0840] 2-chlorotrityl resin (500mg, 0.775mmol) is swollen 15 minutes with 10mL DCM in 50mL Falcon pipe, 5-[(9H-fluorenes-9-ylmethoxy) carbonyl]-5-azaspiro [2.4] heptane-1-carboxylic acid (281mg, 0.775mmol) is dissolved in DCM, DIEA (6 equivalents, 0.81mL) is added, and the solution is added in resin. The solution is purged with argon and vibrates at room temperature overnight. The mixture is filtered and washed with DMF (3x5mL) and DCM (3x5mL). Methanol (5mL) is added, the mixture is vibrated 30 minutes, then filtered. With DMF (3x5mL) and DCM (3x5mL) washing resin. Methanol (5mL) is added again, the mixture is vibrated 30 minutes, then filtered. With DMF (3x5mL) and DCM (3x5mL) washing resin. Based on the determination of the loading procedure described below, the loading amount was determined to be 0.42 mmol / g.

[0841] The resin was then used for automated SPPS on a Symphony X synthesizer from Protein Technologies at a 0.1 mmol scale.

[0842] Determine resin loading:

[0843] To determine the 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 measured photometrically at 301 nm. This directly correlates to the amount of amino acid loaded on the resin.

[0844] 1) Weigh 1-3 mg of resin into a 2 mL Eppendorf tube or similar tube (pay attention to the exact amount)

[0845] 2) Add 1000 μL of 20% piperidine in DMF.

[0846] 3) The mixture was stirred for 30 minutes to cleave the FMOC group.

[0847] 4) Then transfer 100 μL of the supernatant to a quartz glass cuvette and dilute with 900 μL of 20% piperidine / DMF.

[0848] 5) Prepare a 1000 μl piperidine / DMF blank sample in a second cuvette.

[0849] 6) After determining the blank value of the control sample, the extinction of the test sample at λ = 301 nm was then measured in a UV-Vis photometer (Thermo Scientific Evolution 201).

[0850] 7) For higher accuracy, multiple test samples (usually two) can be made; the arithmetic mean of the measured extinction values ​​can then be used for calculations.

[0851] Calculate resin loading :

[0852] Resin loading in mmol / g L 301 Calculated by the following formula:

[0853]

[0854] E = racemic

[0855] ε=Extinction coefficient at 301nm wavelength (7800L / mol*cm)

[0856] m = amount of resin used (g)

[0857] V = sample volume (L)

[0858] D = layer thickness of the cuvette (cm)

[0859] VF = dilution factor (=10)

[0860] Fmoc cleavage:

[0861] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0862] Subsequent coupling of Fmoc amino acids was performed as described in Method A.

[0863] The following solutions were prepared and used during the synthesis:

[0864] 1) Fmoc amino acid: 0.2M (8eq)

[0865] 2) Activator 1: 0.5M DIC in DMF (7.5eq)

[0866] 3) Activator 2: 0.5M Oxyma DMF solution (7.5eq)

[0867] 4) Fmoc deprotection agent: 20% piperidine in DMF solution

[0868] 5) Acetic anhydride in 2 mL DMF (10 equivalents)

[0869] A double coupling is 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 is interrupted and the amino acid is coupled manually (double coupling), but typically with less reagent (3-5 equivalents). After this coupling is complete, the synthesis is typically continued on the synthesizer. If an N-methyl amino acid is added to the sequence, the next amino acid is typically coupled manually as well. All steps are performed at room temperature under nitrogen. Fmoc-Pen and Fmoc-Oic are typically coupled using automated SPPS. Fmoc(N-Me)G is typically coupled manually. Ahx is always coupled manually.

[0870] Fmoc cleavage:

[0871] The resin was swollen and washed with DMF (3 x 3 mL, 10 min). If the resin contained Fmoc, it was removed with 20% piperidine solution (2 x 3 mL, 10 min). The resin was washed with DMF (6 x 3 mL, 30 sec).

[0872] Coupling:

[0873] Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0874] Fmoc cleavage:

[0875] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0876] Coupling:

[0877] Add Fmoc-Pen (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Pen (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0878] Fmoc cleavage:

[0879] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0880] Coupling:

[0881] Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and couple 2 hours under nitrogen bubbling.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and couple 2 hours under nitrogen bubbling.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0882] Fmoc cleavage:

[0883] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0884] Coupling:

[0885] Add Fmoc-Oic (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Oic (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0886] Fmoc cleavage:

[0887] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0888] Coupling:

[0889] Add Fmoc-Pro (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours. Drain solution and wash with DMF (1x3mL, 30 seconds). Repeat coupling step. Add Fmoc-Pro (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours. Drain solution and wash with DMF (6x3mL, 30 seconds).

[0890] Fmoc cleavage:

[0891] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0892] Coupling:

[0893] Add Fmoc-Leu (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Leu (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0894] Fmoc cleavage:

[0895] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0896] Coupling:

[0897] Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0898] Fmoc cleavage:

[0899] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0900] Coupling:

[0901] Add Fmoc-Arg (Pbf) (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3mL, 30 seconds). Repeat the coupling step. Add Fmoc-Arg (Pbf) (4.0mL). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3mL, 30 seconds).

[0902] Fmoc cleavage:

[0903] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0904] Coupling:

[0905] Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Ser (t-Bu) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0906] Fmoc cleavage:

[0907] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0908] Coupling:

[0909] Add Fmoc-Cys (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (1x3 mL, 30 seconds). Repeat the coupling step. Add Fmoc-Cys (Trt) (4.0 mL). Add activator 1 solution (DIC, 1.5 mL) and activator 2 solution (Oxyma, 1.5 mL) and couple under nitrogen bubbling for 2 hours. Drain the solution and wash with DMF (6x3 mL, 30 seconds).

[0910] Fmoc cleavage:

[0911] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0912] Coupling:

[0913] Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (1x3mL, 30 seconds).Repeat coupling step.Add Fmoc-Ile (4.0mL).Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make coupling under nitrogen bubbling, carry out 2 hours.Drain solution and wash with DMF (6x3mL, 30 seconds).Repeat coupling step.

[0914] Fmoc cleavage:

[0915] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0916] Manual coupling during automated SPPS:

[0917] Fmoc-(N-Me) G (0.2M DMF solution, 5 equivalents) is added to the resin. Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under vibration (thermomixer, room temperature) for 2 hours. Filter the solution and wash with DMF (1x3mL, 30 seconds). Repeat the coupling step. Add Fmoc-(N-Me) G (0.2M DMF solution, 5 equivalents). Add activator 1 solution (DIC, 1.5mL) and activator 2 solution (Oxyma, 1.5mL) and make the coupling under vibration (thermomixer, room temperature) for 2 hours. Filter the solution and wash with DMF (6x3mL, 30 seconds).

[0918] Fmoc cleavage:

[0919] The Fmoc protecting group was removed by adding 20% ​​piperidine solution (2 x 3 mL, 10 min) and the resin was washed with DMF (6 x 3 mL, 30 sec).

[0920] If additional amino acids were present in the sequence, they were coupled using the procedure described above.

[0921] Lysis test:

[0922] When performing manual coupling, cleavage is usually performed test To monitor the reaction. test The cleavage mixture 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.

[0923] Full lysis:

[0924] The resin containing the peptide was placed in a syringe and 3.0 mL of lysis 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 collected by filtration and the peptide was precipitated by adding cold ether (-20°C). Under a nitrogen atmosphere, the solution was centrifuged (3000 rpm) in a Falcon tube (60 mL). The ether was decanted and the solid residue was repeatedly washed with cold ether (5 x 10 mL). The solid residue was then dried.

[0925] Disulfide cyclization:

[0926] 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 overnight in a round-bottom flask open to air on an orbital shaker. The solution was then lyophilized to obtain a white powder.

[0927] Column screening for HPLC purification:

[0928] The peptides were dissolved in 5% CH3CN and 95% water. Column screening was performed for each peptide to determine the preparative HPLC method for purification. The following analytical columns were screened.

[0929] Two methods can be used for column screening:

[0930] 1)5-60%ACN_8Min_1mL / min_25℃

[0931] 2)30-85%ACN_8Min_1mL / min_25℃

[0932] Usable column (50mm x ID 4, 6mm) (can also be used as a preparative column) :

[0933] 1) Aeris C18 (Phenomenex)

[0934] 2) X-Bridge C18 (Waters)

[0935] 3)Kinetex C18(Coreshell Material)(Phenomenex)

[0936] 4) YMC triart C18 (can be eluted with 100% water)

[0937] 5)Kinetix Biphenyl(Phenomenex)

[0938] 6) X-Select C18 (positive charge)

[0939] 7)Jupiter Proteo C18 (Phenomenex)

[0940] 8) Luna C18 (Phenomenex)

[0941] For the peptides of the present invention, one of the following five preparative columns was used:

[0942] 1) Column: Phenomenex, Aeris Peptide 5μ XB-C18, AXIA Packed, 21, 2x250mm+Cartridge 5μ

[0943] 2) Column: Phenomenex, Kinetex C18 5μ 21, 5x250mm+Cartridge 5μ

[0944] 3) Column: Phenomenex, Kinetex 5μ Biphenyl 100A, AXIA Packed, 21, 2x250mm+Cartridge 5μ

[0945] 4) Column: YMC Actus triart Prep.C18 12nm, S-10μm 250x20mm+Cartridge 3μm (10x4mm)

[0946] 5) Column: Waters, Xbridge Prep.C18 5μ OBD 19x250mm+Cartridge 10μ.

[0947] After selecting a column, use one of the following methods:

[0948] 1) Method: Gradient 5-60% ACN aqueous solution (0, 10% TFA)

[0949] 2) Method: Gradient 30-85% ACN aqueous solution (0, 10% TFA)

[0950] 3) Enrichment gradient based on column screening results.

[0951] Flow rate 20mL / min

[0952] The combined fractions were analyzed by HPLC (5-95 in 8 min, Chromolith SpeedROD & YMC C18 5-95 in 18 min) and using one of the LC-MS methods described above.

[0953] For Example 164, the crude peptide was dissolved in CH3CN / water (1:1) and purified on a Phenomenex Kinetex column C18 5μ, 21.5×250 mm + 5μ Cartridge, flow rate: 20 mL / min, method: 5-60% ACN in water (each containing 0.10% TFA) over 48 minutes. The combined fractions were lyophilized to provide 29.8 mg (98.3% purity) of Example 164.

[0954] Table 7: Description of materials and conditions used

[0955]

[0956] General method for manual SPPS of Masp peptides (Method C)

[0957] The synthesis of the sequence AIC+SRSLP-(Oic)-I-(Pen)+IPN-OH (Example 418) is representative.

[0958] Synthetic peptides:

[0959] The peptide was synthesized using standard Fmoc chemistry.

[0960] 1) Preparation of resin: To chlorotrityl resin (CTC Resin) (0.3 mmol, 0.3 g, 1.0 mmol / g) was added a solution of Fmoc-Asn(Trt)-OH (0.3 mmol, 0.18 g, 1.0 eq) and DIEA (0.21 mL, 1.2 mmol, 4.0 eq) in DCM (6.0 mL). The mixture was stirred at 20° C. with N2 for 2 hours, then MeOH (0.3 mL) was added and stirred with N2 for another 30 minutes. The resin was washed with DMF (6.0 mL x 3). A solution of 20% piperidine in DMF (6.0 mL) was then added and the mixture was stirred at 20° C. with N2 for 20 minutes. The mixture was then filtered to obtain the resin. The resin was washed with DMF (6.0 mL x 3) and filtered to obtain the resin.

[0961] 2) Coupling: A solution of Fmoc-Pro-OH (0.30 g, 0.9 mmol, 3.0 eq), HBTU (0.32 g, 0.85 mmol, 2.85 eq) and DIEA (1.80 mmol, 0.32 mL, 6.00 eq) in DMF (3.0 mL) was added to the resin and stirred at 20° C. with N 2 for 30 minutes. The resin was then washed with DMF (6.0 mL x 5).

[0962] 3) Deprotection: A 20% piperidine solution in DMF (6.0 mL) was added to the resin, and the mixture was stirred at 20° C. with N 2 for 20 minutes.

[0963] 4) Repeat steps 2 to 3 for all other amino acids:

[0964] Table 8: Description of materials and conditions used

[0965] # Material Coupling reagents Coupling time 1 Fmoc-Asn(Trt)-OH (1.0 eq) DIEA (4.0eq) 2.0 hours 2 Fmoc-Pro-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 3 Fmoc-Ile-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 4 Fmoc-Pen(Trt)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 5 Fmoc-Ile-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 6 Fmoc-Oic-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 7 Fmoc-Pro-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 8 Fmoc-Leu-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 9 Fmoc-Ser(tBu)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 10 Fmoc-Arg(Pbf)-OH (3.0eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 11 Fmoc-Ser(tBu)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 12 Fmoc-Cys(Trt)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 13 Fmoc-Ile-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 14 Fmoc-Ala-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes

[0966] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin (all amino acids except Pr0) and chloranil test (Pro), and the resin was washed 5 times with DMF (5.0 mL).

[0967] Peptide cleavage and purification:

[0968] 1) The resin was washed with methanol (6.0 mL x 5) and vacuum dried to obtain 0.62 g of peptide resin. 6.0 mL of lysis buffer (92.5% TFA / 2.5% EDT / 2.5% TIS / 2.5% H2O) was then added to the flask containing the side-chain protected peptide resin at 20°C, and the mixture was stirred for 2.5 hours.

[0969] 2) The peptide was precipitated with cold tert-butyl methyl ether (50 mL) and centrifuged (3 minutes, 3000 rpm) to obtain a crude solid. The crude peptide precipitate was washed twice more with tert-butyl methyl ether (20.0 mL x 3). The crude peptide was vacuum dried for 2.0 hours to obtain 0.4 g of crude peptide.

[0970] 3) The crude product (0.4 g) was dissolved in CH3CN (150 mL) and water (150 mL). Iodine (I2) (0.1 M methanol solution, 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 aqueous solution, 0.05 mL) was then added dropwise until the yellow color disappeared. The mixture was lyophilized to obtain a crude powder (0.41 g).

[0971] 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) as a white solid and a TFA salt (108.1 mg, 23.0% yield, 97.3% purity by Method W1; 97.4% purity by Method 7). Purification conditions: Peptide dissolved in TFA / H2O (7:3); flow rate 20 mL / min; gradient 12-42% over 60 minutes; retention time = 42 minutes; Luna 25x200 mm, C18 10 μm, Column purification, and then again through Gemini 150x30mm, C18 5μm, Column purification to achieve the desired purity.

[0972] If the C-terminal amide is prepared according to this method, the synthesis begins with Rink Amide MBHA resin (loading typically about 0.4-0.6 mmol / g) and the same steps as above.

[0973] General method for manual SPPS of Masp peptides (Method D)

[0974] If a C-terminal amide is prepared, the synthesis begins with Rink Amide MBHA resin (loading is typically about 0.4-0.6 mmol / g). The synthesis of the sequence PIC+SRS-((tBu)A)-PPI-(Pen)+IPD-NH2 (Example 9) is representative.

[0975] If a C-terminal acid was prepared, the synthesis began with 2-chlorotrityl resin and the first Fmoc amino acid was added as described in Method C.

[0976] Synthetic peptides:

[0977] The peptide was synthesized using standard Fmoc chemistry.

[0978] 1) Preparation of Resin: Rink Amide MBHA resin (0.66 g, 0.30 mmol, loading = 0.45 mmol / g) in DMF (10.0 mL) was stirred at 20° C. with N 2 for 0.5 h. 20% piperidine in DMF (10 mL) was then added and the mixture was stirred at 20° C. with N 2 for 20 min. The mixture was then filtered to obtain the resin. The resin was washed with DMF (10 x 5 mL) and filtered to obtain the resin.

[0979] 2) Coupling: A solution of Fmoc-Asn(Trt)-OH (0.9 mmol, 0.37 g, 3.0 eq), HBTU (0.32 g, 0.85 mmol, 2.95 eq) and DIEA (1.80 mmol, 0.32 mL, 6.00 eq) in DMF (3.0 mL) was added to the resin and stirred at 20° C. with N 2 for 30 minutes. The resin was then washed with DMF (10.0 mL x 3).

[0980] 3) Deprotection: A 20% piperidine solution in DMF (10.0 mL) was added to the resin, and the mixture was stirred at 20° C. with N 2 for 30 minutes.

[0981] 4) Repeat steps 2 to 3 for all other amino acids.

[0982] Table 9: Description of materials and conditions used

[0983] # Material Coupling reagents Coupling time 1 Fmoc-Asn(Trt)-oH (1.0eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 2 Fmoc-Pro-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 3 Fmoc-Ile-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 4 Fmoc-Pen(Trt)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 5 Fmoc-Ile-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 6 Fmoc-Pro-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 7 Fmoc-Pro-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 8 Fmoc-Ala(t-Bu)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 9 Fmoc-Ser(tBu)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 10 Fmoc-Arg(Pbf)-oH (3.0eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 11 Fmoc-Ser(tBu)-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 12 Fmoc-Cys(Trt)-oH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 13 Fmoc-Ile-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes 14 Fmoc-Pro-OH (3.0 eq) HBTU (2.85 eq) DIEA (6.0 eq) 30 minutes

[0984] 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 5 times with DMF (5.0 mL).

[0985] Peptide cleavage and purification:

[0986] 1) The resin was washed with MeOH (6.0 mL x 5) and dried under vacuum to obtain 0.66 g of peptide resin. 6.0 mL of lysis buffer (90% TFA / 5% EDT / 2.5% TIS / 2.5% H2O) was then added to the flask containing the side-chain protected peptide resin at 20°C, and the mixture was stirred for 2.5 hours.

[0987] 2) The peptide was precipitated with cold tert-butyl methyl ether (50 mL) and centrifuged (3 minutes, 3000 rpm) to obtain a crude solid. The crude peptide precipitate was washed three times with tert-butyl methyl ether (20.0 mL x 3). The crude peptide was vacuum dried for 2.0 hours to obtain 0.4 g of crude peptide.

[0988] 3) The crude product (0.4 g) was dissolved in CH3CN (15 mL) and water (15 mL) to a concentration of 0.1 mM. NH4HCO3 solution (1 M) was added to adjust the pH to approximately 8-9. The solution was allowed to vibrate at room temperature for approximately 8 hours. The reaction was monitored by HPLC. After the reaction was complete, the reaction was quenched with acetic acid to adjust the pH to approximately 6. The reaction mixture was then lyophilized and the resulting solid was purified by reverse phase HPLC.

[0989] 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) as a white solid and a TFA salt (202.40 mg, 41.4.0% yield, 93.7% purity by Method W1; 95.1% purity by Method 7). Purification conditions: Peptide dissolved in TFA / H2O (7:3); flow rate 20 mL / min; gradient 12-42% over 60 minutes; retention time = 42 minutes; Luna 25x200 mm, C18 10 μm, Purification on column.

[0990] Alternative general method for manual SPPS of Masp peptides (Method E)

[0991] 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.

[0992] Peptide synthesis:

[0993] The peptide was synthesized using standard Fmoc chemistry.

[0994] The black balls in the chemical structures below represent solid polymer supports used for solid phase polypeptide synthesis (SPPS), such as 2-chlorotrityl resin, Rink amide resin, etc.

[0995] Example 1A

[0996] (2S)-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)(oxa-4-yl)acetic acid (single stereoisomer)

[0997]

[0998] 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) were added to (2S)-amino(oxa-4-yl)acetic acid (950 mg, 5.97 mmol) in acetone / water (15 mL / 10 mL). 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 solution and extracted three more times with dichloromethane. The organic phase was washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and evaporated. The residue was purified by silica gel chromatography (Isolera; 50 g SNAP Ultra; gradient: cyclohexane 50% / ethyl acetate 50% + 1% acetic acid to cyclohexane 20% / ethyl acetate 80% + 1% acetic acid). The appropriate fractions were combined and evaporated. The residue was dissolved in toluene and evaporated twice more, and then the residue was dried in vacuo to give 1.08 g (100% purity, 47% yield) of the target compound.

[0999] LC-MS (Method 9): R t =0.90min; MS (ESIpos): m / z=382[M+H] +

[1000] 1H-NMR (400MHz, 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).

[1001] Example 2A

[1002]

[1003] 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 was added N,N-diisopropylethylamine (19 mL, 110 mmol) and 2-chlorotrityl chloride resin (10.0 g, 13.3 mmol). The reaction mixture was shaken at room temperature overnight. The precipitate was collected by filtration and washed three times with DMF. The collected resin was shaken with dichloromethane / methanol 1:1 for 30 minutes. The resin was collected by filtration and washed three times with methanol and dichloromethane in a rotary vortex. The collected resin was dried under vacuum and used in the next step.

[1004] Example 3A

[1005]

[1006] Example 2A (15.2 g, 6.82 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 15 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the rotary washing was performed 3 times with 200 mL methanol and 200 mL dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1007] Example 4A

[1008]

[1009] Embodiment 3A (11.4g, 5.13mmol) was swollen at room temperature for 5 minutes in 100mL DMF. Add a mixture of N-{[(9H-fluorene-9-yl)methoxy]carbonyl}-S-(trityl)-L-cysteine ​​(6.01g, 10.3mmol), DIC (1.5mL, 10mmol) and Oxyma (1.42g, 10.0mmol) in 50mL DMF, and the reaction mixture was vibrated at room temperature for 2 hours. The resin was collected by filtration and washed 3 times with DMF. The coupling step was repeated overnight. The resin was collected by filtration and washed 3 times with 150mL DMF, then rotated and washed more than 3 times with 150mL methanol and 150mL dichloromethane. The resin collected in vacuo was used for the next step.

[1010] Example 5A

[1011]

[1012] Example 4A (18.9 g, 8.51 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 15 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1013] Example 6A

[1014]

[1015] At room temperature, embodiment 5A (16.3g, 7.32mmol) was swollen in 150mL DMF for 5 minutes. A mixture of N-{[(9H-fluorene-9-yl)methoxy]carbonyl}-L-isoleucine (5.18g, 14.6mmol), DIC (2.2mL, 14mmol) and Oxyma (2.03g, adding 14.3mmol) in 50mL DMF was added, and the reaction mixture was vibrated at room temperature for 2 hours. The resin was collected by filtration and washed 3 times with DMF. The coupling step was repeated overnight. The resin was collected by filtration and washed 3 times with 200mL DMF, then rotated and washed more than 3 times with 200mL methanol and 200mL dichloromethane. The resin collected in vacuo was used for the next step.

[1016] Example 7A

[1017]

[1018] Example 6A (20.0 g, 8.98 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 15 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1019] Example 8A

[1020]

[1021] 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-fluorene-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 vibrated at room temperature for 2 hours. The resin was collected by filtration and washed 3 times with DMF. The coupling step was repeated over the weekend. The resin was collected by filtration and washed 3 times with 200 mL of DMF, then rotated and washed more than 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used for the next step.

[1022] Example 9A

[1023]

[1024] 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, washed three times with DMF, and then rotary washed three more times with 200 mL of methanol and 200 mL of dichloromethane. 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, washed three times with DMF, and then rotary washed three more times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1025] Example 10A

[1026]

[1027] 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-fluorene-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, then rotated and washed three more times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1028] Example 11A

[1029]

[1030] 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 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1031] Example 12A

[1032]

[1033] Embodiment 11A (18.2g, 8.18mmol) was at room temperature swollen 5 minutes in 150mL DMF.Add N-{[(9H-fluorenes-9-yl) methoxy] carbonyl}-L-leucine (11.6g, 32.7mmol), DIC (4.9mL, 32mmol) and Oxyma (4.53g, 31.9mmol) mixture in 50mL DMF, and this reaction mixture is at room temperature vibrated and spent the night.Filter and collect resin, wash 3 times with 200mL DMF, then use 200mL methyl alcohol and 200mL dichloromethane rotary washing more than 3 times.The resin collected in vacuo is used for next step.

[1034] Example 13A

[1035]

[1036] Example 12A (20.9 g, 9.42 mmol) in 150 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1037] Example 14A

[1038]

[1039] Embodiment 13A (18.0g, 8.11mmol) was at room temperature swollen 5 minutes in 150mL DMF.Add the mixture of O-tert-butyl-N-{[(9H-fluorenes-9-yl) methoxy] carbonyl}-L-serine (12.4g, 32.4mmol), DIC (4.9mL, 32mmol) and Oxyma (4.49g, 31.6mmol) in 50mL DMF, and this reaction mixture is at room temperature vibrated and spent the night.Filter and collect resin, wash 3 times with 200mL DMF, then use 200mL methyl alcohol and 200mL dichloromethane rotary washing more than 3 times.The resin collected in vacuo is used for next step.

[1040] Example 15A

[1041]

[1042] Example 14A (21.4 g, 9.64 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1043] Example 16A

[1044]

[1045] Example 15A (18.9 g, 8.51 mmol) was swollen in 150 mL DMF at room temperature for 5 minutes. N in 50 mL DMF was added. 2 -{[(9H-fluoren-9-yl)methoxy]carbonyl}-N 5 To the mixture of 2-[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 rotational washes with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1046] Example 17A

[1047]

[1048] Example 16A (23.8 g, 10.7 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1049] Example 18A

[1050]

[1051] Embodiment 17A (22.1g, 9.95mmol) was at room temperature swollen 5 minutes in 150mL DMF.Add the mixture of O-tert-butyl-N-{[(9H-fluorenes-9-yl) methoxy] carbonyl}-L-serine (15.3g, 39.8mmol), DIC (6.0mL, 39mmol) and Oxyma (5.51g, 38.8mmol) in 50mL DMF, and the reaction mixture is at room temperature vibrated and spent the night.Filter and collect resin, wash 3 times with 200mL DMF, then use 200mL methyl alcohol and 200mL dichloromethane rotary washing more than 3 times.The resin collected in vacuo is used for next step.

[1052] Example 19A

[1053]

[1054] Example 18A (24.7 g, 11.1 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1055] Example 20A

[1056]

[1057] Embodiment 19A (22.2g, 9.99mmol) was at room temperature swollen 5 minutes in 150mL DMF.Add N-{[(9H-fluorenes-9-yl) methoxy] carbonyl}-S-(trityl)-L-cysteine ​​(23.4g, 40.0mmol), DIC (6.0mL, 39mmol) and Oxyma (5.54g, 39.0mmol) mixture in 50mL DMF, and this reaction mixture is at room temperature vibrated and spent the night.Filter and collect resin, with 200mL DMF washing 3 times, use 200mL methyl alcohol and 200mL dichloromethane rotary washing more than 3 times then.The resin collected in vacuo is used for next step.

[1058] Example 21A

[1059]

[1060] Example 20A (24.5 g, 11.0 mmol) in 200 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with 200 mL of methanol and 200 mL of dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1061] Example 22A

[1062]

[1063] Embodiment 21A (22.3g, 10.0mmol) was at room temperature swollen 5 minutes in 150mL DMF.Add N-{[(9H-fluorenes-9-yl) methoxy] carbonyl}-L-isoleucine (14.2g, 40.1mmol), DIC (6.0mL, 39mmol) and Oxyma (5.55g adds 39.0mmol) mixture in 50mL DMF, and this reaction mixture is at room temperature vibrated over the weekend.Filter and collect resin, wash 3 times with 200mL DMF, then use 200mL methyl alcohol and 200mL dichloromethane rotary washing more than 3 times.The resin collected in vacuo is used for next step.

[1064] Example 23A

[1065]

[1066] Example 22A (1.00 g, 250 μmol) in 7.5 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated, and the collected resin was rotated and washed 3 times with methanol and dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1067] Example 24A

[1068]

[1069] Example 23A (1.00 g, 250 μmol) was swollen at room temperature for 5 minutes in 5 mL of DMF. A mixture of (2S)-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)(oxa-4-yl)acetic acid (381 mg, 1.00 mmol), DIC (150 μL, 980 μmol) and Oxyma (139 mg, 975 μmol) was added in 1 mL of DMF, and the reaction mixture was shaken over the weekend. The resin was collected by filtration and washed 3 times with DMF, then rotated with methanol and dichloromethane for more than 3 times. The collected resin was dried under vacuum and used for the next step.

[1070] Example 25A

[1071]

[1072] Example 24A (1.00 g, 250 μmol) in 7.5 mL DMF / piperidine 4:1 was shaken at room temperature for 30 minutes. The resin was collected by filtration and washed 3 times with DMF. These two steps were repeated and the collected resin was rotated and washed 3 times with methanol and dichloromethane. The collected resin was dried under vacuum and used in the next step.

[1073] Example 26A

[1074] (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)-butyl-2-yl]-8-(3-carbamimidamidopropyl)-5,11-bis(hydroxy)- [1-H-indole-2-carbonyl]amino]-3-methylpentanoyl]amino]-3-methylpentanoyl]amino]-3-sulfanylpropionyl]amino]-3-methylpentanoic acid (single stereoisomer)

[1075]

[1076] To Example 25A (1.00 g, 250 μmol) was added a cleavage mixture 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 filtrates were 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 use in the next step.

[1077] Example 184

[1078] ((2S)-2[(Amino)-2-(tetrahydro-2H-pyran-4-yl)]acetic acid)-IC+SRSLP-(Oic)-IC+I-OH (Example 184)

[1079] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-18-({N-[(2S)-2-amino-2-(oxa-4-yl)acetyl]-L-isoleucyl}amino)-26-[(2S)-butan-2-yl]-12-(3-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)- 6-(2-Methylpropyl)-5,8,11,14,17,25,28,35-octaoxotriacontanohydro-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucine (single stereoisomer)

[1080]

[1081] To Example 26A (425 mg, 304 μmol) was added 850 mL of 0.1 M aqueous ammonium bicarbonate (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%).

[1082] General Procedure F: Conversion of Peptide TFA Salts to HCl Salts

[1083] The synthesis of AIC+SRS-((tBu)A)-PPI-((N-Me)C)+IPD-NH2(HCl salt) (Example 30) is representative.

[1084] Automatic Ion Exchange Station steps:

[1085] Peristaltic pump (Rotarus volume 50) from Hirschmann, tubing: Tygon 2001 (inner diameter (ID) 0.64 mm)

[1086] set up:

[1087] Water washing: running time 1200s; 80min -1 ; 1 cycle (volume 35mL)

[1088] Peptide sample cycle: Run time 1200s; 80min -1 ; 1 cycle (volume 70mL)

[1089] Washing with H2O (or % ACN in water): running time 1200s; 80min -1 ; 1 cycle (volume 35mL)

[1090] Amberlite IRA 410 (HCl form) was used. 1500 mg of the resin was placed in 2 filter cartridges and washed with deionized water (10 times).

[1091] The peptide dissolved in 3 mL of 5% ACN / H2O solution was loaded onto the column and circulated through the column 10 times. The column was washed with water and the solution was collected into a Falcon tube and lyophilized.

[1092] 72.83 mg of the desired peptide as HCl salt were obtained: LC-MS (>99%); ion chromatography analysis: 3.7 wt% Cl- (1.57 eq Cl-), <1 wt% TFA.

[1093] The ion exchange process can also be performed using the following scheme:

[1094] Amberlite IRA 410 resin (HCl form) (1-2 g) was placed in a 10 mL glass-syringe (1 g of IRA 410 resin was required for 100 mg of peptide).

[1095] 1) Wash the resin with water (10 times x 3 mL)

[1096] 2) Wash the resin with 5% ACN aqueous solution (1 time x 3 mL)

[1097] 3) Dissolve the peptide in 5% ACN aqueous solution

[1098] 4) Add the peptide to the syringe and circulate the solution through the column 10-20 times. Collect the eluate into a Falcon tube.

[1099] 5) Wash the resin with 5% ACN aqueous solution (10 times x 3 mL); add this solution to the solution in the Falkon tube

[1100] 6) The combined solution was lyophilized.

[1101] General Method FA: Conversion of Peptide TFA Salts to Other Salts

[1102] Other salt forms:

[1103] The chloride counterion can be exchanged for another counterion by repeatedly passing a solution of the desired salt form (e.g., sodium acetate) through the column and then washing the column repeatedly with water. The peptide is then loaded and the above procedure is followed. Thus, peptide acetate, tartrate, citrate, and lactate salts are prepared. The following three examples are representative:

[1104] Example 249

[1105] Sequence: ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH2(acetate) (Example 249)

[1106] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methyl- 1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolinamide, acetate

[1107] IRA 410 chlorinated resin (12 g) was placed in an empty 20 mL Biotage column and washed with 5% ACN-water solution (10 times). The column was connected to an automated ion exchange station. The resin was washed with 20 column volumes of 1 M NaOH solution (2 x 1800 s at a speed of 80 min). -1 ). The column was washed with water until the pH of the eluent was < pH 9 (10 column volumes). The column was then eluted with 1 M acetic acid solution (10 column volumes) and then washed with water until the pH of the eluent was > 5 (approximately 10 column volumes). The peptide (N-Me)GIC+SRSLP-(Oic)-I-Pen+IP-NH2 (TFA salt) (1200 mg) was loaded onto the chromatography column in 12 mL of 5% acetonitrile / water and circulated through the column 10 times according to General Method F. The resin was then washed 10 times with 5% acetonitrile / water solution and the eluent was collected in a 50 mL Falcon tube. The parameters for this method (scale) were set as follows:

[1108] Water washing: running time 3085s; 80min -1 ; 1 cycle (90 mL volume)

[1109] Peptide sample cycle: run time 3085s; 80min -1 ; 1 cycle (180 mL volume)

[1110] Washing with H2O (or with % ACN in water): running time 3085s; 80min -1 ; 1 cycle (90 mL volume)

[1111] The combined eluates were lyophilized to obtain 1120 mg (100% purity, 97% yield) of the target acetate.

[1112] Ion chromatography (chloride content): <1% chloride

[1113] Ion chromatography (TFA content): <1% TFA

[1114] Ion chromatography (acetate content): 7.1% acetate = 1.84 equivalents

[1115] LC-MS (MCW-TOF-AQ-YMC-18 minutes): Rt = 7.74 minutes; MS (ESIpos): m / z = 725 [M+2H] +

[1116] Example 264

[1117] Sequence: ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH2(L-tartrate) (Example 264)

[1118] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl)-5,8,11,1 4,17,25,28,35-Octahedotriacontanohydro-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolinamide (2R,3R)-2,3-dihydroxybutanedioic acid (L-(+) tartaric acid) salt

[1119] IRA 410 chlorinated resin (12.5 g) was placed in an empty 20 mL Biotage column and washed with 5% ACN-water solution (10 times). The column was connected to an automated ion exchange station. 20 column volumes of 1 M NaOH solution (2 x 1800 s at a speed of 80 min) were added. -1) was washed with water until the pH of the eluent was < pH 9 (10 column volumes). The column was then eluted with 1M L-tartaric acid solution (10 column volumes) and then washed with water until the pH of the eluent was > 5 (approximately 10 column volumes). The peptide ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH2 (TFA salt) (1200 mg) was loaded onto the column in 12 mL of 5% acetonitrile / water and circulated through the column 10 times according to General Procedure F. The resin was then washed 10 times with 5% acetonitrile / water solution and the eluent was collected in a 50 mL Falcon tube. The parameters used for this method were set as follows:

[1120] Water washing: running time 3085s; 80min -1 ; 1 cycle (90 mL volume)

[1121] Peptide sample cycle: run time 3085s; 80min -1 ; 1 cycle (180 mL volume)

[1122] Washing with H2O (or % ACN in water): running time 3085s; 80min -1 ; 1 cycle (90 mL volume)

[1123] The combined eluates were lyophilized to give 1170 mg (100% purity, 91% yield) of the desired L-tartrate salt.

[1124] Ion chromatography (chloride content): <1% chloride

[1125] Ion chromatography (TFA content): <1% TFA

[1126] Ion chromatography (acetate content): 13.4% acetate = 1.49 equivalents

[1127] LC-MS (MCW-TOF-AQ-YMC-18min): Rt=7.62min; MS (ESIpos): m / z=725[M+2H] +

[1128] Example 376

[1129] Sequence: ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-NH2(citrate) (Example 376)

[1130] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl)-5,8,1 1,14,17,25,28,35-Octaoxotriacontanohydro-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolinamide, 2-hydroxypropane-1,2,3-tricarboxylic acid (citrate) salt.

[1131]

[1132] IRA 410 chlorinated resin (26 g) was placed in an empty 20 mL Biotage column and washed with 5% ACN-water solution (10 times). The column was connected to an automated ion exchange station. 20 column volumes of 1 M NaOH solution (2 x 1800 s at a speed of 80 min) were added. -1 ) washed resin. The column was washed with water until the pH of the eluent was pH 9 (10 column volumes). The column was then eluted with 1M citric acid solution (10 column volumes) and then washed with water until the pH of the eluent was >5 (approximately 10 column volumes). Peptide (N-Me) GIC + SRSLP-(Oic) -I-Pen + IP-NH2 (TFA salt) (1200 mg) was loaded onto the column in 12 mL 5% acetonitrile / water and circulated through the column 10 times according to General Method F. The resin was then washed 10 times with 5% acetonitrile / water solution and the eluent was collected in a 50 mL Falcon tube. The parameters for this method were set to:

[1133] Water washing: running time 3085s; 80min -1 ; 1 cycle (90 mL volume)

[1134] Peptide sample cycle: run time 3085s; 80min -1 ; 1 cycle (180 mL volume)

[1135] Washing with H2O (or % ACN in water): running time 3085s; 80min -1 ; 1 cycle (90 mL volume)

[1136] The combined eluates were lyophilized to obtain 1370 mg (100% purity, 97% yield) of the target L-tartrate.

[1137] Ion chromatography (chloride content): <1% chloride

[1138] Ion chromatography (TFA content): <1% TFA

[1139] Ion chromatography (acetate content): 19.0% acetate = 1.77 equivalents

[1140] LC-MS (MCW-TOF-AQ-YMC-18 minutes): Rt = 7.6 minutes; MS (ESIpos): m / z = 725 [M+2H] +

[1141] Those skilled in the art will recognize that although ion chromatography stoichiometry is not always stoichiometric (eg, 1:2 or 1:1), it does not take into account water content, which in rare cases can be measured at 8-15%.

[1142] General Method G: Preparation of salt-free form

[1143] The preparation of AIC+SRSLP-(Oic)-I-(Pen)+IPD-NH2 (salt-free form) (Example 27) is representative.

[1144] 5 g of the TFA salt (Example 75) was purified by reverse phase HPLC using an acetonitrile-water gradient at 70°C without an acid modifier. The desired fractions were combined and lyophilized. 2.2 g of the salt-free form (Example 27) was obtained; LC-MS (purity >99%); ion chromatography (<1% TFA)

[1145] General Method H: Large-Scale Synthesis

[1146] The synthesis of AIC+SRS-((tBu)A)-PPI-((N-Me)C)+IPD-NH2 (Example 48) is representative

[1147]

[1148] The peptide was synthesized using standard Fmoc chemistry.

[1149] 1) DMF was added to a vessel containing MBHA resin (180 mmol) and the resin was allowed to swell for 2 hours.

[1150] 2) Add the Fmoc-Rink amide linker and mix for 30 seconds, then add activation buffer and mix using N2 bubbling for approximately 1 hour.

[1151] 3) The solution was drained from the resin and the resin was washed with DMF wash solution (30 seconds x 3 times).

[1152] 4) Add 20% piperidine / DMF solution and mix by bubbling nitrogen for 30 minutes.

[1153] 5) Drain the solution and wash the resin with DMF (5 times).

[1154] 6) Add the Fmoc-amino acid solution and mix for 30 seconds, then add the activation buffer and mix using N2 bubbling for about 1 hour.

[1155] 7) Repeat steps 3 to 6 for each amino acid coupling.

[1156] Table 10: Description of materials and conditions used

[1157]

[1158]

[1159] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[1160] Peptide cleavage and purification:

[1161] 1) Lysis buffer (92.5% TFA: 2.5% EDT: 2.5% TIS: 2.5% H2O) was added to the flask containing the side-chain protected peptide at room temperature, and the mixture was stirred for 2 hours.

[1162] 2) The peptide was precipitated with cold isopropyl ether and centrifuged (at 5000 rpm for 2 minutes).

[1163] 3) The peptide was washed twice more with isopropyl ether.

[1164] 4) Vacuum dry the crude peptide for 2 hours.

[1165] Disulfide formation

[1166] To a solution of the crude linear peptide (15 g, 9.92 mmol, 1.0 eq) (20 batches) in H O / I (3 L 2:1) was added NH HCO (7.84 g, 99.2 mmol, 10 eq). 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 preparative HPLC to give the desired Example 48 (41.8 g, 95.8%, 13.3% yield) as a white solid, a total of 156.8 g.

[1167] Purification conditions

[1168] Apparatus: Shimadzu 10A; Peptide was dissolved in DMF / H2O. Mobile phase: A(H2O (0.075% TFA in water), BEtOH); Gradient: 10-50% over 60 minutes. Retention time: 47 minutes. Column: Luna, C18, 10 μm, 100A, 25 cm x 50 mm; Flow rate: 80 mL / min; Wavelength: 220 / 254 nm; Oven temperature: room temperature.

[1169] Method I: Preparation of ((N-Me)A)-IC+SRSLP-(Oic)-I-(Pen)+IP-((N-Benzyl)D)-NH2 (Example 462)

[1170] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidopropyl)-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-octaoxotriacontanoic acid-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazaheteronadecanoic acid[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolyl-N- 4 -Benzyl-L-asparagine

[1171]

[1172] The peptide was prepared on a 0.3 mmol scale on Rink Amide MBHA resin (loadings are typically about 0.4-0.6 mmol / g). 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 constructed using the steps described in Method C. After the sequence was complete, the 2-OPP protecting group was removed using a solution of 1% TFA in DCM (30 minutes). N-benzylamine (3 equivalents), DIC (3 equivalents), and Oxyma (3 equivalents) were added and the mixture was shaken for 2 hours. The peptide was cleaved to form a disulfide bond and the peptide was purified according to Method C.

[1173] General Procedure J. Preparation of ((N-Me)G)-IC+SRSLP-(Oic)-I-(Pen)+IP-(*Dap)-OH (Example 467)

[1174] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,29aS,33aS,35aS)-26-[(2S)-butan-2-yl]-12-(3-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-18-[(N-methylglycyl-L-isoleucyl)amino]-6-(2-methylpropyl)-5,8, 11,14,17,25,28,35-Octaoxotriacontanohydro-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-N-[(2S)-2-amino-2-carboxyethyl]-L-prolinamide.

[1175]

[1176] The peptide was prepared on 2-chlorotrityl resin according to Method C. BOC-Dap(Fmoc)-OH (CAS 122235-70-5) was added to the resin according to Method C. The peptide was constructed and purified according to Method C.

[1177] General Procedure K. Preparation of (2-(Morpholino)acetyl)-IC + SRS-((tBu)A)-PPI-(Pen) + IPD-NH2 (Example 283)

[1178] Example 27A

[1179] 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-α-asparagine

[1180]

[1181] On Rinkamide ChemMatrix resin, prepare sequence IC+SRS-((tBu) A)-PPI-(Pen)+IPD-NH2 12 times on 0.1mmol scale according to method A.In the peptide that comprises resin, add N, N '-diisopropylcarbodiimide (31 μ L, 200 μ mol), ethyl (hydroxyimino) cyanoacetate (28.4mg, 200 μ mol) and bromoacetic acid (14 μ L, 200 μ mol), and by mixture at room temperature shaken overnight on hot mixing shaker in 5mL DMF.Filter resin, with DMF (6x5mL) washing, with DCM (6x5mL) washing, then vacuum drying is used for next step.

[1182] Example 283

[1183] N-[(5aS,11S,14S,17S,20S,23R,28R,31S,33aS)-31-[(2S)-butan-2-yl]-17-(3-carbamimidoylaminopropyl)-11-(2,2-dimethylpropyl)-14,20-bis(hydroxymethyl)-27,27-dimethyl-23-({N-[(morpholin-4-yl)acetyl]-L-isoleucine}- ... amino)-5,10,13,16,19,22,30,33-octaoxooctacosanehydro-1H,5H,10H-dipyrrolo[2,1-j:2′,1′-m][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosanetacosa-28-carbonyl]-L-isoleucyl-L-prolyl-L-α-asparagine

[1184]

[1185] In the embodiment 27a (200mg, purity 74%, 77.7 μmol) in NMP (10mL), add morpholine (34 μl, 390 μmol) and suspension is vibrated at 37 ℃ in a thermomixer. Solid resin is collected by filtration, washed twice with 10mL DMF and 10mL dichloromethane respectively. Residue is handled with TFA / EDT / thioethanol (thionaisol) (90: 3: 7) and at room temperature vibrated 2 hours. The reaction mixture is handled with cold ether (-10 ℃), and then the suspension is centrifuged under nitrogen and washed 3 times (3x30mL) with cold ether. In the residue, 0.1M aqueous ammonium bicarbonate solution (400mL) is added, and the reaction mixture is vibrated in a round-bottom flask connected to air overnight. The solution is freeze-dried and according to method A, the lyophilized powder is purified to provide 20.4mg (purity 98%, productive rate 16%) of embodiment 283.

[1186] Example 300 (2-hydroxyacetyl)-IC+SRS-((tBu)A)-PPI-(Pen)+IPD-NH2 was isolated as a by-product.

[1187] General Procedure L. Preparation of Sodium and Choline Salt Forms

[1188] These salts are prepared from the peptides in their salt-free form. The following two examples represent a general approach:

[1189] Example 431

[1190] 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-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,1 1,14,17,25,28,35-Octahedraoxotriacontano[1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-N-[(1S)-1,2-dicarboxyethyl]-L-prolinamide, sodium salt

[1191]

[1192] Sequence: AIC+SRSLP-(Oic)-I-(Pen)+IPD-OH (sodium salt) (Example 431)

[1193] To a solution of AIC+SRSLP-(Oic)-I-(Pen)+IPD-OH (salt-free form) (100 mg) in 20 mL of acetonitrile / water 1:1 was added 1.3 mL of 0.1 M sodium hydroxide solution. The mixture was dried in vacuo to afford 109 mg (104% yield, 99% purity) of the title compound.

[1194] LC-MS (MCW-TOF-AQ-YMC-18 minutes): Rt = 8.62 minutes; MS (ESIpos): m / z = 783 [M+2H]2+

[1195] Ion chromatography (sodium content): 2.9% sodium = 2.03 equivalents

[1196] Example 432

[1197] 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-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,11,14,17,25,2 8,35-octaoxotriacontanohydro-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-N-[(1S)-1,2-dicarboxylic acid ethyl]-L-prolinamide, 2-hydroxy-N,N,N-trimethylethane-1-ammonium)(choline) salt

[1198]

[1199] To a solution of AIC+SRSLP-(Oic)-I-(Pen)+IPD-OH (salt-free form) (100 mg) in 20 mL of acetonitrile / water 1:1 was added 511 μL of 0.25 M choline hydroxide solution. The mixture was dried under vacuum to afford 122 mg (105% yield, 99% purity) of the title compound.

[1200] LC-MS (MCW-TOF-AQ-YMC-18min): Rt=8.65 minutes; MS (ESIpos): m / z=783[M+2H]2+

[1201] Ion chromatography (choline content): 11.7% choline = 1.97 equivalents

[1202] Example 13

[1203] AIC+SRSLP-(Oic)-I-(Pen)+IPD-OH (Example 13)

[1204] (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-carbamimidoylaminopropyl)-5,11-bis(hydroxymethyl) [2-(2-methylpropyl)-4,7,10,13,16,19-hexaoxa-14-(sulfanylmethyl)-3,6,9,12,15,18-hexaazaheneicosanoyl]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

[1205]

[1206] synthetic peptides

[1207] Peptides were synthesized using standard Fmoc chemistry.

[1208] 1) Preparation of resin: Fmoc-Asp(OtBu)-OH (15.0mmol, 6.17g, 1.0eq) and DIEA (10.4mL, 60.0mmol, 4.0eq) in DCM (300mL) were added to CTC resin (15.0mmol, 15.0g, 1.0mmol / g). The mixture was stirred at 20°C with N2 for 2 hours. Methanol (15.0mL) was then added and the resin was stirred for another 30 minutes with N2. The resin was washed with DMF (300mL x3). A solution of 20% piperidine in DMF (300mL) was then added and the mixture was stirred at 20°C with N2 for 20 minutes. The mixture was then filtered to obtain the resin. The resin was washed with DMF (300mL x3) and filtered to obtain the resin.

[1209] 2) Coupling: Fmoc-Pro-OH (15.2 g, 45.0 mmol, 3.0 eq), HBTU (16.2 g, 42.8 mmol, 2.85 eq) and DIEA (90.0 mmol, 15.6 mL, 6.00 eq) in DMF (100 mL) were added to the resin, and the resin was stirred at 20° C. with N 2 for 30 minutes. The resin was then washed with DMF (300 mL x 5).

[1210] 3) Deprotection: 20% piperidine solution in DMF (300 mL) was added to the resin and stirred at 20° C. under N 2 for 20 minutes.

[1211] 4) Repeat steps 2 and 3 for all other amino acids:

[1212] Table 11: Reagents used in Example 13

[1213]

[1214]

[1215] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was tested by ninhydrin (all amino acids except Pro) and chloranil (Pr o ) was monitored, and the resin was washed 5 times with DMF (300 mL).

[1216] HBTU, DIEA supplier: Suzhou Highfine Biotech Co., Ltd.

[1217] Peptide cleavage and purification

[1218] 5) The resin was washed with methanol (300 mL x 5) and dried under vacuum to obtain 41.5 g of peptide resin. 400 mL of cleavage buffer (92.5% TFA / 2.5% 3-mercaptopropionic acid / 2.5% TIS / 2.5% H2O) was then added to the flask containing the side-chain protected peptide resin at 20°C. The mixture was stirred for 2.5 h and then filtered.

[1219] 6) The peptide was precipitated with cold tert-butyl methyl ether (4000 mL) and centrifuged (3000 rpm for 3 min) to obtain a crude solid peptide. The crude peptide precipitate was washed three more times with tert-butyl methyl ether (1000 mL x 3). The crude peptide was then vacuum dried for 2.0 hours to obtain 21.8 g of crude peptide.

[1220] 7) The crude peptide (21.8 g) was dissolved in CH3CN (6.5 L) and water (6.5 L). I2 (0.1 M in MeOH, 55 mL) was added at 20°C until a 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 obtain a crude powder (22.7 g).

[1221] 8) The crude peptide was purified by preparative HPLC (conditions: A: 0.075% TFA aqueous solution B: CH3CN), and the fractions containing the product were collected and lyophilized to obtain the target peptide (SEQ ID 13); (4305.1 mg, 18.3% yield, 99.1% by C18 RP-HPLC column (Gemini C18 5 μm 150x4.6mm); 97.8% passed C18 RP-HPLC column (Discovery BIO 5m 150 x 4.6 mm), TFA salt) as a white solid.

[1222] Table 12: Purification conditions

[1223]

[1224]

[1225] LC-MS: Rt = 1.369 min; 1564 [M+2H] +

[1226] Rt = 8.72 min (purity 99.1%) Gemini C18 5 μm 150x4.6mm

[1227] Rt=9.42min(97.8%)Discovery BIO Wide Porei C185μm 150x4.6mm

[1228] Example 29 (HCl salt) can be prepared from Example 13 (TFA salt) using General Procedure F; Example 5 (salt-free form) can be prepared using General Procedure G. The preparations of Example 431 (sodium salt) and Example 432 (choline salt) from Example 5 are shown below as Examples 431 and 432, respectively.

[1229] Example 237

[1230] AIC+SRSL-(L-dehydroproline)-(Oic)-I-(Pen)+IPD-OH (Example 237)

[1231] N-[(6S,9S,12S,15S,18R,23R,26S,28aS,35aS)-18-[(L-alanyl-L-isoleucyl)amino]-26-[(2S)-butan-2-yl]-12-(3-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8,11,14,17,25,28,35-octaoxa-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-Triacontanyl-3H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolyl-L-aspartic acid

[1232]

[1233] The peptide (3.12 g) was prepared according to Method C. The analytical data of the peptide are shown in Tables 16 and 17.

[1234] Example 142

[1235] AIC+SRSL-(Pro-D2)-(Oic)-I-(Pen)+IPD-OH (Example 142)

[1236] 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-carbamimidoylaminopropyl)-9,15-bis(hydroxymethyl)-22,22-dimethyl-6-(2-methylpropyl)-5,8, 11,14,17,25,28,35-Octoxa(1,2-2H2)triacontanohydro-1H,5H,22H-pyrrolo[2′,1′:13,14][1,2,5,8,11,14,17,20,23,26]dithiooctaazacyclononacosa[11,10-a]indole-23-carbonyl]-L-isoleucyl-L-prolyl-L-aspartic acid

[1237]

[1238] The peptide from Example 237 (30 mg, 19.2 mmol) was dissolved in methanol-d4 (3 mL) (99.8% purity D). Rhodium black (CAS 7440-16-6, ACBR, Product No. AB25746) (10 mol%, 1.9 μmol) was added to the solution in an M-Braun glove box under argon. The 8 mL reaction flask mixture was placed in an MTP pressure reactor block on a Chemspeed Swing XL platform and sealed in an M-Braun glove box (Chemspeed Technologies AG, 8, 4414 Fillinsdorf, Switzerland, chemspeed@chemspeed.com). At room temperature and 3 bar pressure of deuterium gas, the reaction was shaken at an orbital speed of 650 rpm for 16 h overnight. The pressure device was purged with argon and the bottle was taken out from the reactor device. Methanol-d4 was evaporated on a rotary evaporator, the resulting solid was dissolved in water (2 mL) and stirred for 1 h to exchange the exchangeable deuterium atom with hydrogen. After 1 hour, acetonitrile was added and the product was analyzed by reverse phase HPLC [column: Phenomenex Aeris Peptide 5 μ XB-C18, AXIA Packed, 21.2x250 mm + Cartridge 5 μ, flow rate: 20 mL / min, enrichment gradient using H2O / ACN and 0.1% TFA as acidic modifier. The product was purified by FOK30-40: 5% acetonitrile at 0 min, ramping to 30% ACN at 0-13 min, gently ramping to 40% ACN at 13-39 min, isocratic 40% acetonitrile at 39-46 min. The chromatographic fractions containing the product were analyzed by analytical HPLC (enriched gradient) and LC-MS and then combined accordingly to provide two fractions of pure product: 11.4 mg (90% purity, 34% yield) and 4.2 mg (99% purity, 14% yield). Analytical data for this peptide are provided in Table 16.

[1239] The product peptide was digested with trypsin and then reacted 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 produces two fragments, AICSR-OH (accurate mass 548.2741, found 548.283 [M+H]+) and SL-(Pro-D2)-(Oic)-I-(Pen)+IPD-OH (accurate mass 1037.5800, found 1038.587 [M+H]+), and the position of the deuterium atom is verified by using the fragments 838.47amu ((Pro-D2)-(Oic)-I-(Pen)+IPD-OH) and 738.40amu SL-((Oic)-I-(Pen)+IPD-OH).

[1240] Example 28A

[1241] (2S,3aS,6aS)-Octahydrocyclopenta[b]pyrrole-2-carboxylic acid-hydrochloride (1 / 1) (single stereoisomer)

[1242]

[1243] 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-hydrochloride (1 / 1) (1.00 g, 3.55 mmol). Hydrogen gas (1 atm) was added to the reaction flask and the reaction mixture was stirred overnight at room temperature under a hydrogen atmosphere. The reaction mixture was filtered through celite and the filter cake was washed with methanol. The filtrate was evaporated and dried under vacuum to give 698 mg of the title compound.

[1244] (2S,3aS,6aS)-Octahydrocyclopenta[b]pyrrole-2-carboxylic acid is also commercially available in salt-free form.

[1245] Example 29A

[1246] (2S,3aS,6aS)-1-{[(9H-fluoren-9-yl)methoxy]carbonyl}octahydrocyclopenta[b]pyrrole-2-carboxylic acid (single stereoisomer)

[1247]

[1248] To (2S,3aS,6aS)-octahydrocyclopenta[b]pyrrole-2-carboxylic acid-hydrochloride (1 / 1) (680 mg, 3.55 mmol) in 12 mL of water was added sodium bicarbonate (2.98 g, 35.5 mmol) and 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. The reaction mixture was stirred at room temperature overnight. 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 solution, 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 100:2-100:5). Appropriate fractions were combined, evaporated and dried in vacuo to give 1.27 g (99% purity, 94% yield) of the title compound.

[1249] LC-MS (Method 10): Rt = 2.08 min; MS (ESI pos): m / z = 378 [M+H] +

[1250] 1H-NMR (400MHz, 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).

[1251] Example 30A

[1252] (1R,3S,5R)-2-Azabicyclo[3.1.0]hexane-3-carboxylic acid (single stereoisomer)

[1253]

[1254] (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 4M 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.

[1255] Example 31A

[1256] (1R,3S,5R)-2-{[(9H-fluoren-9-yl)methoxy]carbonyl}-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (single stereoisomer)

[1257]

[1258] 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-fluorene-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 at room temperature overnight. The reaction mixture was treated with water and extracted twice with ethyl acetate. The organic phase was washed with saturated sodium chloride aqueous solution, dried over magnesium sulfate, filtered, and then concentrated. The crude product was purified by preparative reverse phase HPLC (chromatographic column: Reprosil; C18; 10 μm; 125×30 mm; eluent A: CAN, eluent B: water containing 0.1% formic acid), and the fractions containing the product were combined and lyophilized to give 88.0 mg (96% purity, 11% yield) of the target compound.

[1259] LC-MS (Method 11): Rt = 1.32 min; MS (ESI pos): m / z = 372 [M + Na] +

[1260] 1H-NMR (400MHz, DMSO-d6) δ [ppm]: -0.149 (1.15), -0.008 (10.73), 0.008 (9.1 3), 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).

[1261] This amino acid is used in Fmoc SPPS. This fmoc amino acid is also commercially available.

[1262] Example 32A

[1263] (2R,4R)-4-[(2S)-But-2-yl]-2-tert-butyl-5-oxo-1,3-oxazolidine-3-carboxylic acid benzyl ester (single stereoisomer)

[1264] To a suspension of D-isoleucine (8.06g, 61.4mmol) in 120mL ethanol, a sodium hydroxide solution made from 2.45g sodium hydroxide in 11mL water was added. The reaction mixture was stirred at room temperature for 90min, then evaporated. The residue was treated with 195mL pentane, followed by addition of 2,2-dimethylpropanal (10mL, 92mmol). The reaction mixture was stirred at 45°C for 24h using a Dean-Stark trap (trap) connected 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 165mL dichloromethane and cooled to 0°C. Benzyl chloroformate (Benzylcarbonochloridate) (13mL, 92mmol) was added and the reaction mixture was stirred at 0-4°C for one week. 4-dimethylaminopyridine (375mg, 3.1mmol) and 65mL water were added to the reaction mixture, the reaction mixture was brought to room temperature, then stirred at room temperature overnight. The reaction mixture was treated with ethyl acetate several times and extracted. The combined organic phases were washed twice with 10% aqueous citric acid solution and then twice with saturated aqueous sodium bicarbonate solution. The aqueous phase was back-extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, 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 in vacuo to yield 8.71 g of the title compound.

[1265] Example 33A

[1266] (2R,4R)-4-[(2S)-But-2-yl]-2-tert-butyl-4-methyl-5-oxo-1,3-oxazolidine-3-carboxylic acid benzyl ester (single stereoisomer)

[1267]

[1268] 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 h. Methyl iodide (2.3 mL, 37 mmol) was added dropwise to the reaction mixture and the reaction mixture was stirred at room temperature for 2.5 h. Saturated aqueous ammonium chloride solution (20 mL) was added to the reaction mixture, the reaction mixture was allowed to reach room temperature, and the product was extracted into ethyl acetate. The combined organic phases were washed with saturated aqueous sodium bicarbonate solution, 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 100:3-100:7). The product containing fractions were combined, evaporated and then dried in vacuo to give 3.2 g of the title compound.

[1269] Example 34A

[1270] N-[(Benzyloxy)carbonyl]-2-methyl-D-alloisoleucine (single stereoisomer)

[1271]

[1272] To a solution of (2R,4R)-4-[(2S)-butan-2-yl]-2-tert-butyl-4-methyl-5-oxo-1,3-oxazolidine-3-carboxylic acid benzyl ester (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 at reflux for 2 h. 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 sodium chloride aqueous solution, dried over magnesium sulfate, filtered, evaporated, and then dried in vacuo to give 2.77 g of the title compound.

[1273] Example 35A

[1274] 2-Methyl-D-alloisoleucine (single stereoisomer)

[1275]

[1276] The reaction was carried out under an argon atmosphere. Palladium on carbon (390 mg, 10%) was added to N-[(benzyloxy)carbonyl]-2-methyl-D-isoleucine (Example 34A, 2.57 g, 9.20 mmol) in 190 mL of methanol. The reaction mixture was stirred at room temperature and atmospheric pressure under a hydrogen atmosphere for 4 hours. The reaction mixture was filtered through celite and the filtrate was concentrated and then dried in vacuo to provide 1.32 g (98% yield) of the title compound.

[1277] Example 36A

[1278] N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-2-methyl-D-alloisoleucine (single stereoisomer)

[1279]

[1280] To a solution of 2-methyl-D-isoleucine (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 at room temperature overnight. The reaction mixture was treated with MTBE and extracted. The aqueous phase was acidified with concentrated hydrochloric acid (pH 1) 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 100:3-100:5). The appropriate fractions were combined, evaporated, and then dried under vacuum.

[1281] The MTBE phase was subsequently 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 each time. The combined organic phases were dried over magnesium sulfate, filtered, evaporated, and then dried in vacuo. The combined crude product yield was 2.53 g.

[1282] The residue was purified by silica gel chromatography (Isolera, column: 50 g SNAP Ultra; eluent: dichloromethane / methanol 97:3-97:5). The product-containing fractions were combined, evaporated, and then dried in vacuo to give 967.4 mg of the title compound. The product was further dissolved in water / acetonitrile and lyophilized before use.

[1283] LC-MS (Method 10): Rt = 2.10 min; MS (ESI pos): m / z = 368.1857 [M+H] +

[1284] This amino acid is used for Fmoc SPPS.

[1285] Example 37A

[1286] (6S)-1,1-Difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (mixture of stereoisomers)

[1287]

[1288] 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 h. The reaction mixture was concentrated and dried under vacuum for the next step.

[1289] Example 38A

[1290] (6S)-5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (mixture of stereoisomers)

[1291]

[1292] To a solution of (6S)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (mixture of stereoisomers) (Example 37A, 1.60 g, 9.03 mmol) in water (13 mL) was added sodium bicarbonate (7.59 g, 90.3 mmol) 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 at room temperature overnight. 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 solution, dried over magnesium sulfate, filtered, evaporated and dried in vacuo to afford 3.31 g (92% yield) of the title compound.

[1293] LC-MS (Method 10): Rt = 1.99 min; MS (ESI pos): m / z = 400 [M+H] +

[1294] 1H-NMR (400MHz, 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(0.7 8), 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).

[1295] Example 39A

[1296] (6S)-5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (Enantiomer 1) and (Enantiomer 2)

[1297]

[1298] The diastereomeric mixture was separated using a THAR supercritical fluid chromatography Prep 200 instrument using a Chiralpcel AD-H (SFC) 5 μm, 250x30 mm column; eluent: CO2 / isopropanol (83:17); pressure: 135 bar; eluent temperature: 38°C; zyklon temperature: 40°C; zyklon pressure: 24 bar; flow rate: 100 g / min; detection: UV 210am; injection volume 0.4 mL / injection; under normal operating conditions, 1 g / min of CO2 is approximately equivalent to 1 mL / min.

[1299] Sequence settings: Cycle time = 11.2 minutes

[1300] Fraction 1 (Enantiomer 1) was collected between 5.85 min and 8.0 min.

[1301] Fraction 2 (Enantiomer 2) was collected between 8.4 min and 10.25 min.

[1302] The fractions were concentrated.

[1303] Fractions were analyzed using analytical SFC-MS (Agilent, column: Chiralpak AD-3 3 μm 100×4.6 mm, eluent: CO 2 / isopropanol (85:15); BPR pressure: 130 bar; BPR temperature: 60° C.; column temperature: 40° C.; flow rate: 3 mL / min; UV 210 nm).

[1304] Fraction 1 (rt = 2.081 min) was designated as added "(Enantiomer 1)".

[1305] Fraction 2 (rt = 2.838 min) was designated as "(Enantiomer 2)" added.

[1306] The stereochemistry of the two enantiomers was not specified.

[1307] Example 40A

[1308] (3R*,6S)-5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (single enantiomer, Enantiomer 1, stereochemistry at position 3 not defined)

[1309]

[1310] 2,015 g of the title compound was obtained from chiral SFC preparative purification, 100% ee

[1311] LC-MS (Method 10): Rt = 1.99 min; MS (ESI pos): m / z = 400 [M+H] +

[1312] 1 H-NMR (400MHz, 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(0.4 1), 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).

[1313] This amino acid is used for Fmoc SPPS.

[1314] Example 41A

[1315] (3S*,6S)-5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (single enantiomer, Enantiomer 2, stereochemistry of the cyclopropyl center not specified)

[1316]

[1317] Chiral SFC preparative purification gave 1.041 g of the title compound, 94.58% ee

[1318] LC-MS (Method 10): Rt = 1.93 min; MS (ESI pos): m / z = 400 [M+H] +

[1319] 1H-NMR (400MHz, DMSO-d6) δ [ppm]: -0.008(1.19), 0.008(1.31), 1.031(0.55), 1.047(0.56), 1.285(0.60), 1.583(1.53) , 1.604(4.87), 1.627(5.45), 1.650(3.25), 1.673(2.78), 1.682(3.76), 1.704(3.11), 1.718(0.95), 1.915(3.17), 1.94 9(3.55), 1.979(3.13), 2.012(3.33), 2.073(3.85), 2.526(1.60), 2.590(1.44), 2.626(1.61), 2.649(0.89), 2.659(0.80), 3.471(2.04), 3.498(5.50), 3.527(3.26), 3.561(7.96), 3.570(4.68), 3.598(0.84), 4.167(0.94), 4.183(2.75), 4 .199(6.46), 4.223(7.75), 4.238(9.60), 4.258(5.95), 4.277(4.06), 4.290(3.98), 4.306(3.80), 4.319(4.63), 4.340(4.17), 4.346(4.47), 4.354(2.34), 4.367(3.96), 4.477(3.66), 4.497(3.66), 7.300(1.97), 7.318(8.34), 7.334(11.7 0), 7.337(12.42), 7.352(6.35), 7.355(6.41), 7.407(9.62), 7.425(16.00), 7.444(7.11), 7.633(6.46), 7.651(10.65), 7.666(6.62), 7.683(4.36), 7.883(9.71), 7.893(10.32), 7.901(9.57), 7.912(8.99), 12.802(0.79), 12.984(0.67).

[1320] This amino acid is used for Fmoc SPPS.

[1321] Example 42A

[1322] 3-[4-(tert-Butoxycarbonyl)-1H-1,2,3-triazol-1-yl]-N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-alanine (single stereoisomer)

[1323]

[1324] To a mixture of 3-azido-N-{[(9H-fluoren-9-yl)methoxy]carbonyl}-L-alanine (1.00 g, 2.84 mmol) in 27 mL of water / tert-butanol / dichloromethane (1:1:1) was added a solution of tert-butyl prop-2-ynoate (580 μL, 4.3 mmol), copper(II) sulfate pentahydrate (709 mg, 2.84 mmol), and sodium ascorbate (1.41 g, 7.10 mmol) in 14 mL of water. The reaction mixture was stirred at room temperature overnight. The reaction mixture was treated with water, acidified with dilute citric acid, and extracted three times with dichloromethane. The organic phase was dried over sodium sulfate, filtered, and evaporated. The residue was dissolved in acetonitrile / water and separated into two fractions by preparative HPLC (column: Chromatorex C18 10 μm 125×40 mm; water-acetonitrile containing 0.05% TFA - gradient: 25% acetonitrile from 0 to 2 min, 25-75% acetonitrile from 2 to 22 min, 75% acetonitrile from 23 to 26 min, back to 25% acetonitrile from 26 to 28 min; flow rate 100 mL / min). The product-containing fractions were combined, evaporated and dried in vacuo to give 1.02 g (100% purity, 75% yield) of the target compound.

[1325] LC-MS (Method 11): Rt=1.30 min; MS (ESI neg): m / z=477 [MH]-

[1326] This amino acid is used for Fmoc SPPS.

[1327] Example 43A

[1328] 5-Azaspiro[2.4]heptane-1-carboxylic acid-trifluoroacetic acid (1 / 1)

[1329]

[1330] To a solution of 5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-1-carboxylic acid (1.00 g, 4.14 mmol) in 25 mL of dichloromethane was added TFA (960 μL, 12 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was evaporated and dried under vacuum to give 0.80 g (76% yield) of the title compound.

[1331] LC-MS (Method 9): Rt = 0.18 min; MS (ESI pos): m / z = 142 [M+H] +

[1332] Example 44A

[1333] 5-{[(9H-fluoren-9-yl)methoxy]carbonyl}-5-azaspiro[2.4]heptane-1-carboxylic acid

[1334]

[1335] To a solution of 5-azaspiro [2.4] heptane-1-carboxylic acid-trifluoroacetic acid (1 / 1) (Example 43A, 994 mg, 3.90 mmol) in a mixture of 5 mL of acetonitrile and 5 mL of water was added 1-({[(9H-fluorenes-9-yl) methoxy] carbonyl} oxy) pyrrolidine-2,5-dione (2.63 g, 7.79 mmol) and triethylamine (2.2 mL, 16 mmol). The reaction mixture was stirred at room temperature overnight. The reaction mixture was treated with water, acidified to pH 4 with 1 M hydrochloric acid, and then extracted 3 times with dichloromethane. The combined organic phase was dried over sodium sulfate, filtered, and evaporated. The residue was ground with acetonitrile, then filtered to collect the precipitate and washed with acetonitrile. The filtrate was purified in triplicate by preparative HPLC (column: Chromatorex C18 10 μm 125×40 mm; water-acetonitrile containing 0.05% TFA - gradient: 25% acetonitrile from 0 to 2 min, 25% to 75% acetonitrile from 2 to 22 min, 75% acetonitrile from 23 to 26 min, back to 25% acetonitrile from 26 to 28 min; flow rate 100 mL / min). Appropriate fractions were combined, evaporated, and dried in vacuo to give 1.16 g (95% purity, 78% yield) of the target compound.

[1336] LC-MS (Method 9): Rt = 0.99 min; MS (ESI pos): m / z = 364 [M+H] +

[1337] This amino acid is used for Fmoc SPPS. ...

Claims

1. A peptide-containing compound or a pharmaceutically acceptable salt thereof, wherein the peptide is 2. A peptide-containing compound according to claim 1, or a pharmaceutically acceptable salt thereof, which acts as a MASP-1 and / or MASP-2 inhibitor and / or inhibits C3 deposition.

3. A method for preparing a peptide-containing compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, said method using solid phase polypeptide synthesis.

4. The peptide-containing compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof for use in the prevention and / or treatment of the following diseases: cardiovascular and cardiopulmonary diseases, shock, cerebral and renal sequelae of sepsis, ischemia and / or reperfusion-related injuries, diseases of the blood and hematopoietic organs and the immune system, sequelae of diabetes, or urogenital system diseases, wherein the urogenital system diseases are selected from acute renal failure, acute renal injury, renal ischemia and infarction, allergies in the case of hemodialysis and hemodiafiltration, cystitis, inflammatory diseases of the prostate and endometriosis.

5. The peptide-containing compound or a pharmaceutically acceptable salt thereof according to claim 4, wherein the acute kidney injury is surgery-related acute kidney injury, sepsis-related acute kidney injury, contrast agent- and chemotherapy-induced acute kidney injury.

6. The peptide-containing compound or a pharmaceutically acceptable salt thereof according to claim 4, wherein the cystitis is radiation cystitis.

7. A pharmaceutical composition comprising at least one peptide-containing compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, and one or more inert, non-toxic, pharmaceutically suitable excipients.

8. A pharmaceutical composition comprising at least one peptide-containing compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, and one or more other active ingredients selected from the group consisting of phosphodiesterase inhibitors, guanylate cyclase stimulators or activators, IP receptor agonists, mineralocorticoid receptor antagonists, diuretics, PPAR-γ agonists, PPAR-δ agonists, corticosteroids, active ingredients that reduce organ damage under oxidative stress, compounds that inhibit pathways that induce cell death and apoptosis, 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 heparin derivatives, and coagulation factor XI inhibitors.

9. A pharmaceutical composition according to claim 7 or 8 for the prevention and / or treatment of the following diseases: cardiovascular and cardiopulmonary diseases, shock, cerebral and renal sequelae of sepsis, ischemia and / or reperfusion-related injuries, diseases of the blood and hematopoietic organs and the immune system, sequelae of diabetes, or urogenital system diseases, wherein the urogenital system diseases are selected from acute renal failure, acute kidney injury, renal ischemia and infarction, allergies in the case of hemodialysis and hemodiafiltration, cystitis, inflammatory diseases of the prostate and endometriosis.

10. The pharmaceutical composition according to claim 9, wherein the acute kidney injury is surgery-related acute kidney injury, sepsis-related acute kidney injury, contrast agent- and chemotherapy-induced acute kidney injury.

11. The pharmaceutical composition according to claim 9, wherein the cystitis is radiation cystitis.

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