Apelin peptidic macrocycles and uses thereof

Abstract

An apelin peptidic macrocyclic compound is provided. Specifically, provided is a compound of formula (II), or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof. Also provided is a method of treating a cardiovascular disease in a subject in need thereof using an apelin peptidic macrocyclic compound of the present disclosure, the method comprising administering to the subject an effective amount of the compound. # imgabs0 #

Description

[0001] Cross-reference to related applications

[0002] This application is PCT application serial number PCT / CA2022 / 05*, filed on December 15, 2022, and published in English under PCT Article 21(2), which itself claims the benefit of U.S. Provisional Application Serial No. 63 / 290,394, filed on December 16, 2021. All of the above documents are incorporated herein by reference in their entirety.

[0003] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0004] not applicable Technical Field

[0005] The present disclosure relates to apelinergic macrocycles and uses thereof. More specifically, the present disclosure relates to apelinergic macrocycles derived from apelin-13, apelin-17 and Elabela.

[0006] References to sequence listings

[0007] Pursuant to 37 CFR 1.821(c), a sequence listing created on December 15, 2022 and having a size of 80 kilobytes is filed herewith as an ASCII-compatible text file named G14692-0087.xml. The contents of the above-mentioned file named G14692-0087.xml are incorporated herein by reference in their entirety. Background Art

[0008] APJ receptors are regulators of the cardiovascular system and metabolic activity. Two endogenous ligands, apelin and ELABELA, bind to APJ receptors with nanomolar affinity and stimulate cardiac contraction, thereby reducing resistance to peripheral vascular disease, endothelial cell proliferation and new blood vessel formation. Its protective effect has been demonstrated in pathological models of myocardial ischemia, cerebral ischemia, diabetes, pulmonary hypertension, sepsis and neuropathic pain. However, these endogenous peptides have poor plasma stability, which limits their use. Most studies have shown that apelin and ELABELA must be administered by continuous infusion to maintain their therapeutic efficacy.

[0009] Adrenergic drug analogs are used as standard treatment for cardiac dysfunction associated with sepsis. However, they are not always effective and cause multiple side effects, such as myocardial or peripheral ischemia. Resistance to treatment in pulmonary hypertension is a recurring problem. Existing drugs have shown variable effectiveness, and the prognosis of this disease remains poor. Analgesics other than opioids are also needed to avoid dose escalation and side effects.

[0010] Improved apelin peptidergic analogs are needed.

[0011] This specification refers to various documents, the entire contents of which are incorporated herein by reference in their entirety. Summary of the invention

[0012] The present disclosure provides novel macrocyclic analogs of apelin 13, apelin 17, and Elabela (apelinergic macrocycle). Macrocyclization has been applied to apelin isoforms and Elabela, resulting in molecules with significantly improved stability (plasma half-life between 5 and 24 hours compared to 24-30 minutes for apelin-13 and Elabela). Affinity was measured by displacement of radiolabeled ligand (Nle75, Tyr77) [125I]]-Pyr-apelin 13, which indicated that some macrocyclic peptides exhibited similar affinity to apelin (Ki 0.2-5.7nM) for APJ.

[0013] In specific embodiments, the macrocyclic analogs disclosed herein have a reduced size (33% reduction in molecular mass for apelin-13) while maintaining good affinity to the receptor (Ki 0.8-5 nM for apelin-13, Ki 0.8 nM).

[0014] In specific embodiments, the macrocyclic apelin analog produces cardiovascular effects comparable to those of the endogenous ligand.

[0015] More specifically, according to the present disclosure, the following items and items' are provided:

[0016] Item 1. A compound of any one of formula (I) to (VIII) or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0017] Item 2. The compound according to Item 1, which is any one of Compounds 3-4, 9-29, 35-46, 62-70, 72-79, 84 and 89-94 of Tables I to III.

[0018] Item 3. A pharmaceutical composition comprising the compound, stereoisomer, mixture, pharmaceutically acceptable salt, ester or solvate according to Item 1 or 2, and at least one pharmaceutically acceptable carrier or excipient.

[0019] Item 4. A method of using a compound of any one of Formulae (I) to (IV) to treat cardiovascular disease in a subject in need thereof, the method comprising administering an effective amount of the compound to the subject.

[0020] Item 5. The method according to Item 4, wherein the compound is any one of Compounds 3-4, 9-29 and 35-46.

[0021] Item 6. The method according to Item 5, wherein the compound is compound 42 or 43.

[0022] Item 1. A compound of formula (II):

[0023] (II)

[0024] in:

[0025] X1 does not exist or is X7-X8, where

[0026] X7 is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, a natural amino acid, a synthetic amino acid, the side chain of the natural amino acid, the synthetic amino acid is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally substituted with one or two (C3-C wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents, each of which is independently, for example, halogen, amine, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are N, O or S; and

[0027] X8 is absent or is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH2, -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, The heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidino group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S;

[0028] Y is absent or is NH2-, Ac-NH-, guanidine or H;

[0029] A is -(CH2)n-; -(CH2)nNH=C(NH2)N-CH2-CH=CH- (preferably allyl-glycine or Nα-allyl-arginine), where n is 2, 3 or 4; or -CH=CH-(CH2)m-, where m is 0, 1 or 2;

[0030] B does not exist or

[0031] wherein R is O, P, m-alkyl, halogen or nitro, and n is 1, 2 or 3; wherein R is H, C3-C7 alkyl, benzyl or arylalkyl, and n is 1, 2 or 3; wherein n is 1, 2, 3 or 4, and m is 0 or 1; or, wherein X9 is CH or N;

[0032] X2 and X3 are each independently absent or a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH2, -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally substituted with at least one amino or guanidino group; wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl groups; and wherein the heteroatoms in the heteroalkyl, the heteroaryl or the heterocycloalkyl group are 1, 2 or 3 N, O or S;

[0033] X4 is a natural or unnatural amino acid with a positively charged or uncharged side chain;

[0034] X5 is Gly, Phe, Leu, He, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn); and

[0035] X6 is X10-X11-X12, where

[0036] X10 is any natural amino acid; or a synthetic amino acid, the side chain of which is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH2, -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is substituted with one or more substituents, wherein each substituent is independently For example, halogen, amino, guanidino, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p' is 0 to 5; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p is 0 to 5. In a specific embodiment, X10 is not Ala. In a more specific embodiment, X10 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Phe, Tic ((S)-N-Fmoc-tetrahydroisoquinoline-3-carboxylic acid), Tyr, 1Nal, 2Nal, TyrOBn, cypTyr(OBn), dcypTyr(OBn), cypTyr(OCyp), cypTyr(OPr), D-1Nal, D-2Nal, D-TyrOBn or D-Tyr;

[0037] X11 is absent or is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X11 is absent or is Pro; and

[0038] X12 does not exist or is Phe,

[0039] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0040] Item '2. A compound according to Item '1, wherein:

[0041] X2 and X3 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4; and / or

[0042] X10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently -OH, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p' is 0 to 5,

[0043] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0044] Item '3. A compound according to Item '1, wherein:

[0045] X2 and X3 are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg, His, Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), or α-methylphenylalanine;

[0046] X4 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn);

[0047] X5 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn); and / or

[0048] X10 is X10 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Phe, Tic ((S)-N-Fmoc-tetrahydroisoquinoline-3-carboxylic acid), Tyr, 1Nal, 2Nal, TyrOBn, cypTyr(OBn), dcypTyr(OBn), cypTyr(OCyp), cypTyr(OPr), D-1Nal, D-2Nal, D-TyrOBn, or D-Tyr,

[0049] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0050] Item '4. A compound according to Item '3, wherein:

[0051] X2 and X3 are each independently Lys, Arg, hArg, Nle, Leu, Phe or Cha;

[0052] X4 is Gly; and / or

[0053] X5 is Pro,

[0054] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0055] Item '5. A compound according to any one of Items '1 to 4, wherein:

[0056] X1 does not exist;

[0057] Y is NH2, Ac-NH-, guanidine or H;

[0058] A is -CH=CH-(CH2)m-, wherein m is 0, 1 or 2;

[0059] B is absent, wherein R is O, P, m-alkyl, halogen or nitro, and n is 1, 2 or 3, wherein X9 is CH or N; and / or

[0060] X10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently -OH, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p' is 0 to 5,

[0061] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0062] Item '6. A compound according to Item '5, wherein X10 is Nle or D-1Nal or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0063] 2. A compound according to any one of items 1 to 4, wherein:

[0064] X1 is X7-X8; and / or

[0065] Y does not exist,

[0066] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0067] Item '7. A compound according to Item '7, wherein:

[0068] X1 is X7-X8, and X8 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4,

[0069] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0070] Item '8. A compound according to Item '7 or 8, wherein

[0071] A is -(CH2)n- or -CH=CH-(CH2)m-, wherein m is 0, 1 or 2,

[0072] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0073] Item '9. A compound of any one of formula (I) to (VIII) or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0074] Item '10. The compound according to Item '9, which is any one of compounds 3-4, 9-29, 35-46, 62-70, 72-79, 84 and 89-94, preferably any one of compounds 11, 13, 15-16, 18-20 and 42-44:

[0075]

[0076]

[0077] or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0078] Item '11. The compound according to Item '10, which is any one of Compounds 13-25, 27-29, 35-37 and 42-45, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0079] Item '12. A pharmaceutical composition comprising a compound, stereoisomer, mixture, pharmaceutically acceptable salt, ester or solvate according to any one of Items '1 to 11, and at least one pharmaceutically acceptable carrier or excipient.

[0080] Item '13. A method for treating cardiovascular disease in a subject in need thereof using a compound of any one of Formulas (I) to (IV) or a stereoisomer or mixture thereof or a pharmaceutically acceptable salt, ester or solvate thereof, the method comprising administering an effective amount of the compound to the subject.

[0081] Item '14. The method according to Item '13, wherein the compound is any one of compounds 3-4, 9-29 and 35-46 as defined in Item '10, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0082] Item '15. The method according to Item '13, wherein the compound has formula (II) as defined in any one of Items '1 to 8.

[0083] Item '16. The method according to Item '15, wherein the compound is any one of compounds 13-25, 27-29, 35, 36-37 and 42-45 as defined in Item '10, preferably any one of compounds 13, 15-16, 18-20, 23 and 42-44, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0084] Item '17. The method according to Item '16, wherein the compound is compound 42 or 43, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

[0085] Other objects, advantages and features of the present disclosure will become more apparent from a reading of the following non-limiting description of specific embodiments of the present disclosure, which is given by way of example only with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0086] Figure 1 : Structures of Apel3 and compound 97. The cyclization positions Pro3 and His7 are indicated and circled.

[0087] Figure 2A -B: Macrocyclic Ape13 analogs with various linkers ( Figure 2A ) and various non-natural residues ( Figure 2B ).

[0088] Figure 3A -B: Macrocyclization by ring-closing metathesis to produce precursors of 97, 16, 18 and 19. 97 (SEQ ID NO: 87-89) ( Figure 3A ) and 16, 18 and 19 (SEQ ID NOs: 90-92) ( Figure 3B )'s linear and cyclic precursors.

[0089] Figure 4 :N α -Fmoc-(N π -allyl)-L-histidine-OH. (a) MeOH, HOBt, EDC, DCM, room temperature, overnight, 73%; (b) i. Tf2O (1.1 eq.), DIPEA (1.2 eq.), allyl alcohol (1.1 eq.), -78°C, for 10 min, and room temperature, overnight; ii. TFA, TIPS, DCM, room temperature, 2 h, 70%, for 2 steps; (c) HCl 2M, dioxane-water, reflux, overnight, 77%.

[0090] Figure 5 : Synthesis of peptides containing Fmoc-Alnb. Linear precursors of compounds 15, 18 and 19.

[0091] Figure 6 :N α-Synthesis of Fmoc-cypTyr(OR)-OH analogs. (a) Phosphoric acid 85%, cyclopentanol, 100°C, overnight; (b) SOCl2, MeOH, room temperature, overnight, 26%, for 2 steps a and b; (c) Boc2O, NaHCO3, THF-water (1:1), room temperature, 1 hour, 84%; (d) RBr, K2CO3, ACN, reflux, overnight, yields 17a (57%), 17b (54%), 17c (52%); (e) LiOH, THF-water (1:1), room temperature, 3 hours, yields 113 (96%), 114 (100%), 115 (93%); (f) i. TFA-DCM (1:1), room temperature, 2 hours; ii. Fmoc-Cl, NaHCO3, THF-water (2:1), room temperature, 2 hours, yields 116 (68%), 117 (41%), 118 (71%).

[0092] Figure 7 : N-terminally truncated analogs of 97, 15 and 16, namely compounds 20-23 and 24-28.

[0093] Figure 8 : Nle11 (compound 29) was substituted by natural and unnatural amino acids to generate compounds 34-45.

[0094] Fig. 9A -B: Synthesis scheme of illustrative compounds of Formula VI (eg, compound 79 of Table III, AM03-68).

[0095] Fig.10 : Synthetic schemes for illustrative compounds of Formula VII (eg, compounds 75, 77-78 and 89-93 of Table III).

[0096] Fig.11A -B: Synthetic schemes for illustrative compounds of Formula VIII (eg, compounds 72-73 of Table III).

[0097] Fig.12 :Ape13 macrocyclic analogs in Gα i1 , Gα 12 Concentration-response curves on the β-arrestin 2 pathway. Monitoring ligand-triggered G protein Gα activation using a BRET-based G protein dissociation assay i1 Engagement of (A) (Galés et al., 2006). Ligand-induced β-arrestin2 recruitment using BRET-based β-arrestin2 recruitment assay (B) (Galés et al., 2006). Each group represents the average of at least three independent experiments and is expressed as mean ± SEM.

[0098] Fig.13: In vivo pharmacokinetic profiles of macrocycles 42 and 43 in male Sprague-Dawley rats (n=3). Compounds were administered intravenously at 3 mg / kg and their concentrations in blood samples were quantified by LC / MS-MS.

[0099] Fig.14 : Hypotensive effects of compounds 15, 20, 29, 42 and 43 in anesthetized male Sprague-Dawley rats. Follow-up depicts changes in blood pressure after receiving bolus injections (intravenous) of compounds 15, 20, 29, 39, 42 and 43 or Apel3 via the jugular vein at two doses (65 nmol / kg and 19.6 nmol / kg) (n=4-6 per group).

[0100] Fig.15A -B: Effect of macroloops 42 and 43 on left ventricular fractional shortening ( Fig.15A ) and cardiac output ( Fig. 15B ) (n=5 for all groups). *p<0.05, **p<0.01 vs. time-matched NS (unstimulated) using one-way ANOVA test. DETAILED DESCRIPTION

[0101] Compounds of the Disclosure

[0102] The present disclosure provides novel apelinergic compounds, which include apelin 13 analogs, apelin 17 analogs and elabela analogs.

[0103] Apelin is a peptide hormone that acts as an endogenous ligand for the class A G protein-coupled APJ receptor (Tatemoto et al., 1998; O'Dowd et al., 1993; and Read et al., 2019). As a GPCR target, APJ receptors are known to bind to different G proteins, such as Gα i APJ receptors also signal by recruiting β-arrestin, which is associated with receptor desensitization (Besserer-Offroy et al., 2018; and Gurevich et al., 2019). It is well known that the β-arrestin pathway couples to various effectors and initiates downstream signaling on its own (Gurevich et al., 2019; and Reiter et al., 2012).

[0104] Apelin and elabela are two endogenous peptide ligands for APJ and, despite very different primary sequences, have similar binding potency and signaling profiles (Chng et al., 2013; Pauli et al., 2014; and Murza et al., 2016). Apelin exists in several isomeric forms: apelin-36, apelin-17, apelin-13, [Pyr 1 ]-Apelin-13 and [Pyr 1 ]-Apelin-13 (1-12) Among them, [Pyr 1 ]-Apelin-13 (Ape13) is the major isomer circulating in human plasma and cardiac tissue (Tatemoto et al., 1998; Maguire et al., 2009; Yang et al., 2017; Nyimanu et al., 2019; Zhen et al., 2013).

[0105] Compounds of the Disclosure

[0106] In specific embodiments, the macrocyclic compounds of the present disclosure are developed by cyclization of synthetic peptides (typically made of natural and / or unnatural amino acids) derived from apelin-13 (PyrRPRLSHKGPMPF (SEQ ID NO:47)), apelin-17 (KFRRQRPRLSHKGPMPF (SEQ ID NO:86)), or a fragment of Elabela (PyrRRCMPLHSRVPFP (SEQ ID NO:85)).

[0107] In specific embodiments, the cyclization of the peptide is a side chain to side chain cyclization. In specific embodiments, the cyclization of the synthetic peptide is achieved by the reaction of a ring-closing metathesis of an alkene (-C=C-) (or an alkyne (-C≡C-)) group at the end of each side chain of the N and central amino acid (or acid) moiety, cyclization produces a single carbon-carbon double bond (or a single carbon-carbon triple bond if an alkyne group is used). The macrocycle can then be further modified to replace the double bond by a single bond by palladium-catalyzed hydrogenation. (See, e.g., compound 13).

[0108] In other specific embodiments, cyclization of the peptide is achieved by a macrolactamisation reaction between an amine at the end of the side chain of one of the N-terminal amino acids and a carboxylic acid at the end of the side chain of the amino acid residue used to close the ring, or vice versa.

[0109] In a specific embodiment, the compounds of the present disclosure have any one of Formulas I to VIII, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof. If there is a difference between the name (residue list) and the structure (formula) mentioned herein for the compounds of the present disclosure or a portion thereof, the structure (formula) shall prevail. If there is a difference between the compound described in the sequence table and the name (residue list) and / or structure (formula) mentioned herein for the compounds of the present disclosure or a portion thereof, the name (residue list) and / or structure (formula) shall prevail.

[0110] The reference to an amino acid or an acid as a part of a molecule of the present disclosure herein should be construed as specifying an amino acid or an acid residue. At least one of its ends is connected to another amino acid or an acid to form, for example, a peptide bond, thereby losing a hydroxyl group and / or a hydrogen of an amine group. Therefore, for example, an amino acid or an acid listed in any one of the definitions of X1, X2, X3, X4, X5, and X6 should be construed as corresponding amino acids or acid residues.

[0111] The compounds of the present disclosure have a binding affinity (Ki binding (nM)) for APJ of less than 1000 nM; in specific embodiments, less than 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40, 35, 30, 35, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3 or 2 nM or less than 1 nM. In specific embodiments, the compounds of the present disclosure are any one of Formulas I to VIII or Tables I to III having a binding affinity (Ki binding (nM)) to APJ of less than 1000 nM; in specific embodiments, less than 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40, 35, 30, 35, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3 or 2 nM or less than 1 nM.

[0112] Unless otherwise indicated, the definitions of residues provided herein include both L and D configurations.

[0113] Apelin 13 analogs

[0114] The present disclosure encompasses apelin 13 cyclic analogs, such as those described in Formulas (I)-(IV).

[0115] In one embodiment, the apelin 13 cyclic analogue comprises or consists of the following formula (I):

[0116] X1-X2-Y-[X3-X4-X5-X6-X7-X8-X9-X10-X11-X12]-X13-X14-X15-X16-X17-X18,

[0117] in

[0118] X1 is absent or is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, or is any natural amino acid; or any synthetic amino acid, the side chain of which is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with one or more Two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl are fused; and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amine, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, and wherein the heteroatom in the heteroalkyl, heteroaryl or heterocycloalkyl is N, O or S. In a specific embodiment, the X1 is Pyr or absent;

[0119] X2 and X7 are each independently absent or a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X2 and X7 are each independently absent or an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, wherein p is 0 to 4. In a specific embodiment, X2 and X7 are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg, His or absent. In a more specific embodiment, X2 and X7 are each independently absent or -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4; or are each independently Arg or Lys;

[0120] Y is H, Ac, Ac-NH, -NH2, guanidine or absent,

[0121] X3 and X12 are ring-closed and identical or different, and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, it is a residue carrying a terminal olefin or a free carboxyl or amine function. In a specific embodiment, it is each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α, γ-diamino-butyric acid, Nγ-allyl-α, γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α, γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, at least one or both of X3 and X12 are allylglycine. In a specific embodiment, if X3 is allylglycine, then X12 is not allylglycine.

[0122] X4, X5 and X6 are each independently Ser, Thr, Asn, Gln, Asn-(8-aminocaprylic acid), Trp-(8-aminocaprylic acid) or do not exist. In a specific embodiment, X4, X5 and X6 are each independently Thr, Asn, Asn-(8-aminocaprylic acid), Trp-(8-aminocaprylic acid) or do not exist. In another specific embodiment, they all do not exist.

[0123] X8 is absent or is Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X8 is absent or is Pro.

[0124] X9 is any natural amino acid; or a synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a more specific embodiment, X9 is absent or is any natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a more specific embodiment, X9 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a more specific embodiment, X9 is Nle, Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg or hArg. In a more specific embodiment, X9 is Arg;

[0125] X10 is any natural or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidino, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a more specific embodiment, X10 is Leu, Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine, α-tolylalanine, Ala, Val, Ile. In a more specific embodiment, X10 is Leu;

[0126] X11 is absent or is any natural amino acid; or any synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a more specific embodiment, X11 is Ser or absent;

[0127] X13 is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In one embodiment, X13 is Lys, Orn, Dab, Dap, Arg, -CH2-(CH2)p-guanidine, wherein p is 0 to 4, or His. In one embodiment, X13 is Lys.

[0128] X14 is Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In a specific embodiment, X14 is Gly;

[0129] X15 and X17 are each independently Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X15 and X17 are each independently absent or Pro.

[0130] X16 is any natural amino acid; or a synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X16 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein the aryl is optionally fused with one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl. In a more specific embodiment, X16 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine, α-methylphenylalanine, Phe, Tic ((S)-N-Fmoc-tetrahydroisoquinoline-3-carboxylic acid), Tyr, 1Nal, 2Nal, TyrOBn, cypTyr(OBn), dcypTyr(OBn), cypTyr(OCyp), cypTyr(OPr), D-1Nal, D-2Nal, D-TyrOBn or D-Tyr;

[0131] X18 is absent; or is any natural amino acid; or a synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In one embodiment, X18 is absent or is an amino acid whose side chain is -(CH2)p-(C3-C8)aryl, wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently halogen, amine, -OH, S or -(C1-C6)alkyl. In another embodiment, X18 is Phe or Phe substituted with halogen. In another embodiment, X18 is absent. In another embodiment, X18 is Phe.

[0132] In a specific embodiment of a compound of formula (I), when X3 is allylglycine, X12 is not allylglycine. In a specific embodiment of a compound of formula (I), when X17 and X18 do not exist, X16 is not Ala. In a specific embodiment, the compound of formula (I) is any one of compounds 13-29 and 35-46 of Table I. In other specific embodiments, the compound of formula (I) is any one of compounds 13, 15-16, 18-20, 28 and 42-44 of Table I.

[0133] The present disclosure includes compounds of formula (I), wherein each of X1 to X18 is independently defined using any of the more general or more specific definitions provided above for these residues in formula (I).

[0134] In another specific embodiment, the apelin 13 cyclic analogue comprises or consists of the following formula (II):

[0135]

[0136] Where X1 does not exist or is X7-X8,

[0137] wherein X7 is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, a natural amino acid, a synthetic amino acid, the side chain of the natural amino acid, the synthetic amino acid is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally substituted with one or two (C3 -C8) aryl, (C3-C8) heteroaryl, (C3-C8) cycloalkyl or -(C3-C8) heterocycloalkyl fused; and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amine, -OH, S, -(C1-C6) alkyl, -O-(C1-C6) alkyl, -(CH2)p-(C3-C8) aryl, -O-(CH2)p-(C3-C8) aryl, -(C3-C8) cycloalkyl or -O-(C3-C8) cycloalkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are N, O or S. In a specific embodiment, X7 is Pyr; and

[0138] X8 is absent or is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X8 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a specific embodiment, X8 is Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg, His or absent, preferably Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg or hArg. In a more specific embodiment, X8 is Arg.

[0139] Y is absent or is NH2-, Ac-NH-, guanidine or H;

[0140] A is -(CH2)n-; -(CH2)nNH=C(NH2)N-CH2-CH=CH- (preferably Nα-allyl-arginine), where n is 2, 3 or 4; or -CH=CH-(CH2)m, where m is 0, 1 or 2 (preferably allyl-glycine);

[0141] B does not exist or

[0142] wherein R is O, P, m-alkyl, halogen or nitro, and n is 1, 2 or 3; wherein R is H, C3-C7 alkyl, benzyl or arylalkyl, and n is 1, 2 or 3; wherein n is 1, 2, 3 or 4, and m is 0 or 1; or wherein X9 is CH or N.

[0143] X2 and X3 are each independently absent or a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X2 and X3 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a specific embodiment, X2 and X3 are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg, His, Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine) or α-methylphenylalanine. In a specific embodiment, X2 and X3 are each independently Lys, Arg, hArg, Nle, Leu, Phe or Cha. In a more specific embodiment, X2 and X3 are each independently Arg or Lys.

[0144] X4 is a natural or unnatural amino acid with a positively charged or uncharged side chain. In a specific embodiment, X4 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In a specific embodiment, X4 is Gly;

[0145] X5 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In a specific embodiment, X5 is Pro;

[0146] X6 is X 10 -X 11 -X 12 ,in

[0147] X 10is any natural amino acid; or a synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X 10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently -OH, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p is 0 to 5. In a specific embodiment, X10 is not Ala. In a more specific embodiment, X 10 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Phe, Tic ((S)-N-Fmoc-tetrahydroisoquinoline-3-carboxylic acid), Tyr, 1Nal, 2Nal, TyrOBn, cypTyr(OBn), dcypTyr(OBn), cypTyr(OCyp), cypTyr(OPr), D-1Nal, D-2Nal, D-TyrOBn, or D-Tyr;

[0148] X 11is absent or is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X11 is absent or is Pro; and

[0149] X 12 Does not exist or is Phe.

[0150] In specific embodiments, the compound of formula (II) is any one of compounds 13-25, 27-29, 35-37 and 42-45 of Table I. In other specific embodiments, the compound of formula (II) is any one of compounds 13, 15-16, 18-20, 28 and 42-44 of Table I.

[0151] In a specific embodiment, X1 is Pyr-Arg, Y is -NH-, A is -CH2-CH2-, B is absent, X2 is Arg, X3Lys, X4 is Gly, X5 is Pro, and X6 is Nle-Pro-Phe.

[0152] In another embodiment, X1 is Pyr-Arg, Y is -NH-, A is -CH=CH-, and B is X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle-Pro-Phe.

[0153] In another embodiment, X1 is Pyr-Arg, Y is -NH-, A is -CH=CH-, and B is X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle-Pro-Phe.

[0154] In another embodiment, X1 is Pyr-Arg, Y is -NH-, A is -CH2-CH2-, and B is X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle-Pro-Phe.

[0155] In another embodiment, X1 is Pyr-Arg, Y is -NH-, A is -CH=CH-, and B is X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle-Pro-Phe.

[0156] In another specific embodiment, X1 is H, Y is -NH-, A is -CH=CH-, B is absent, X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle-Pro-Phe.

[0157] In another specific embodiment, X1 is absent, Y is -H, A is -CH=CH-, B is absent, X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle.

[0158] In another specific embodiment, X1 is H, Y is -NH-, A is -CH=CH-, B is absent, X2 is Nle, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle.

[0159] In another specific embodiment, X1 is H, Y is -NH-, A is -CH=CH-, B is absent, X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is Nle.

[0160] In another specific embodiment, X1 is H, Y is -NH-, A is -CH=CH-, B is absent, X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is D-1Nal.

[0161] In another specific embodiment, X1 is H, Y is -NH-, A is -CH=CH-, B is absent, X2 is Arg, X3 is Lys, X4 is Gly, X5 is Pro, and X6 is D-2Nal.

[0162] In a specific embodiment of the compound of formula (II), when A is allylglycine, B is not allylglycine.

[0163] The present disclosure includes compounds of formula (II), wherein each of the variables X1, X2, X3, X4, X5, X6, Y, A and B are independently defined using any of the more general or more specific definitions provided above for these residues in formula (II).

[0164] In another specific embodiment, the apelin 13 cyclic analogue comprises or consists of the following formula (III):

[0165] X1-[X2-X3-X4-X5-X6-X7]-X8-X9-X10-X11-X12-X13-X14-X15,

[0166] wherein X1 is absent or is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, a natural amino acid, a synthetic amino acid, the side chain of the natural amino acid, the synthetic amino acid is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally substituted with one or two ( -(C3-C8)aryl, -(C3-C8)heteroaryl, -(C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl fused; and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amine, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are N, O or S. In a specific embodiment, X1 is Pyr, -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11;

[0167] X2 and X7 are ring-closed and identical or different and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, it is a residue carrying a terminal olefin or a free carboxyl or amine function. In a specific embodiment, it is each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α,γ-diamino-butyric acid, Nγ-allyl-α,γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α,γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, at least one or both of X2 and X7 are allylglycine;

[0168] X3 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X3 is absent or is Pro;

[0169] X4 is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X4 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a specific embodiment, X4 is Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg or His. In a more specific embodiment, X4 is Arg or Lys. In a more specific embodiment, X4 is Arg.

[0170] X5 and X6 are each independently absent or any natural amino acid or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidino, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X5 is Leu and / or X6 is Ser;

[0171] X8 is absent or is any natural amino acid or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X8 is Ser;

[0172] X9 is absent or is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are preferably 1, 2 or 3 N, O or S. In one embodiment, X9 is absent or is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -(CH2)p-imidazole, wherein p is 0 to 4. In one embodiment, X9 is absent or is Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg or His. In a more specific embodiment, X9 is absent or is His;

[0173] X10 is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a more specific embodiment, X10 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a specific embodiment, X10 is Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg or His. In a more specific embodiment, X10 is Lys;

[0174] X11, X12 and X14 are each independently Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X11 is Gly. In one embodiment, X12 and / or X14 are Pro;

[0175] X13 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, preferably Nle;

[0176] X15 is an amino acid whose side chain is -(CH2)p-(C3-C8)aryl, wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently halogen, amine, -OH, S or (C1-C6)alkyl. In a specific embodiment, X15 is Phe or Phe substituted with halogen. In a specific embodiment, X15 is Phe.

[0177] In specific embodiments, the compound of formula (III) is any one of compounds 3 and 4 of Table I.

[0178] The present disclosure includes compounds of formula (III), wherein each of X1 to X15 is independently defined using any of the more general or more specific definitions provided above for these residues in formula (III).

[0179] In another specific embodiment, the apelin 13 cyclic analogue comprises or consists of the following formula (IV):

[0180] X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-[X11-X12-X13-X14-X15],

[0181] wherein Xaa1 is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, a natural amino acid, a synthetic amino acid, the side chain of the natural amino acid, the synthetic amino acid is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with one or two (C and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents, each of which is independently, for example, halogen, amine, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are N, O or S. In a specific embodiment, X1 is Pyr, -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11. In a more specific embodiment, said X1 is Pyr;

[0182] X2, X4, X7 and X8 are each independently a natural or synthetic amino acid, the side chain of which is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X2, X4, X7 and X8 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, wherein p is 0 to 4. In a specific embodiment, X2, X4, X7 and X8 are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg or His. In a more specific embodiment, X2, X4, X7 and X8 are each independently Arg, His or Lys. In a more specific embodiment, X2, X4 and X8 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4; or are each independently Arg or Lys; and / or X7 is an amino acid whose side chain is -(CH2)p-imidazole, wherein p is 0 to 4; or is His;

[0183] X3 is Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X3 is Pro;

[0184] X5 and X6 are each independently any natural amino acid or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X5 is Leu; and / or X6 is Ser;

[0185] X9 is absent or is a natural or unnatural amino acid with a positively charged or uncharged side chain. In a specific embodiment, X4 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In a specific embodiment, X4 is Gly;

[0186] X10 and X12 are each independently absent or Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In a more specific embodiment, X10 is absent. In a specific embodiment, X10 and X12 are each independently absent or Pro;

[0187] X11 and X15 are ring-closed and identical or different and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, they are residues carrying terminal olefins or free carboxyl or amine functions. In a specific embodiment, they are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α,γ-diamino-butyric acid, Nγ-allyl-α,γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α,γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, X12 and X15 are independently allylglycine or D-allylglycine;

[0188] X13 is absent or is any natural amino acid or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidino, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In specific embodiments, X13 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-tolylalanine, preferably Nle.

[0189] X14 is Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X14 is Pro.

[0190] In a specific embodiment of the compound of formula IV, X9 and X10 are absent. In a specific embodiment, if X11 is allylglycine, then X15 is not allylglycine. In a specific embodiment, X11 is not B1 or B2.

[0191] In specific embodiments, the compound of formula (IV) is any one of compounds 9-12 of Table I. In other specific embodiments, the compound of formula (I) is compound 12 of Table I.

[0192] The present disclosure includes compounds of formula (IV), wherein each of X1 to X15 is independently defined using any of the more general or more specific definitions provided above for these residues in formula (IV).

[0193] In specific embodiments of formulas (I) to (IV), at least 4 (or at least 5, 6, 7, 8, 9 or 10) residues (e.g., residues at positions Xn defined above, wherein n is 1 to 18 for formula (I), 1 to 12 for formula (II), or 1 to 18 for formulas (III) and (IV)) correspond to residues in AP-13 (SEQ ID NO: 47), except for the residues of the closed loop that differ from the corresponding residues in AP-13 (SEQ ID NO: 47). For example, compound 3 has at least 10 residues corresponding to residues in AP-13 (SEQ ID NO: 47).

[0194] In other specific embodiments, the compounds of the present disclosure correspond to macrocyclic analogs of Ap13PyrRPRLSHKGPMPF (SEQ ID NO: 47), wherein the compounds differ from Ap13 in at least two substitutions and at least one (or 2, 3, 4, 5, 6, 7 or 8) additional substitutions, deletions and / or insertions at the position of the closed loop. These substitutions, deletions and / or insertions are defined in the various Xn of formulas (I) to (IV) above. The correspondence between these Xn and Ap13 is shown in Table A below, where the "[" and "]" symbols are used to represent the ring closure residues in formulas (I) to (IV) and the positions of the compounds of the present disclosure that satisfy these formulas.

[0195]

[0196] Apelin 17 analogs

[0197] In one embodiment, the apelin 17 cyclic analogue comprises or consists of the following formula (V):

[0198] X1-X2-X3-X4-X5-X6-X7-X8-X9-[X10-X11-X12-X13]-X14-X15-X16-X17,

[0199] Wherein X1 is a natural or synthetic amino acid whose side chain is -R-CH2-(CH2)p-NH 2、-R-CH2-(CH2)p-guanidine, -R-(CH2)p-(C3-C8)cycloalkyl, -R-(CH2)p-(C3-C8)heterocycloalkyl, -R-(CH2)p-(C3-C8)aryl or -R-(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidino group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl groups; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl groups are preferably 1, 2 or 3 N, O or S, preferably N, wherein R is absent or is acetyl. In one embodiment, X1 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In one embodiment, X1 is R-Lys, R-Orn, R-Dab (2,4-diaminobutyric acid), R-Dap (2,3-diaminopropionic acid), R-Arg, R-hArg of R-His, wherein R is absent or is acetyl. In one embodiment, X1 is Ac-Lys;

[0200] X2 is Phe;

[0201] X3, X4, X6, X8, X11 and X12 are each independently a natural or synthetic amino acid, the side chain of which is -(CH2)p-(C3-C8)alkyl, -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X2, X4, X7, X8, X11 and X12 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a specific embodiment, X3, X4, X6, X8, X11 and X12 are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg or His. In a specific embodiment, X3, X4, X6, X8, X11 and X12 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4; or are each independently Arg, His or Lys. In a specific embodiment, X3, X4, X6 and X8 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine, wherein p is 0 to 4; or Arg; and / or X11 is an amino acid whose side chain is -(CH2)p-imidazole, wherein p is 0 to 4; or His; and / or X12 is an amino acid whose side chain is -CH2-(CH2)p-NH2, wherein p is 0 to 4; or X12 is Lys;

[0202] X5 is Gln;

[0203] X7, X14 and X16 are each independently Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In a specific embodiment, at least one, two or all three of X7, X14 and X16 are Pro;

[0204] X9 and X15 are each independently any natural amino acid or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidinyl, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, the heteroaryl or the heterocycloalkyl are 1, 2 or 3 N, O or S. In a more specific embodiment, X9 and X15 are each independently a natural or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5. In a more specific embodiment, X9 and X15 are each independently a natural or synthetic amino acid whose side chain is -(C3-C6)alkyl. In another specific embodiment, X9 and X15 are each independently Nle, Leu, Ala, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine. In a more specific embodiment, X9 is Leu; and / or X15 is Ala or Nle;

[0205] X10 and X13 are ring-closed and identical or different, and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, it is a residue carrying a terminal olefin or a free carboxyl or amine function. In a specific embodiment, it is each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α,γ-diamino-butyric acid, Nγ-allyl-α,γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α,γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, one of X10 and X13 is Glu, and the other is Lys;

[0206] X17 is any natural amino acid or any synthetic amino acid, the side chain of the natural amino acid or the synthetic amino acid is -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl or -(CH2)p-CONH-aryl; or -(CH2)p-CON(aryl)(alkylaryl), wherein p is 0 to 5, and wherein the heteroaryl, the aryl, the cycloalkyl and the heterocycloalkyl are optionally substituted with one or more substituents; wherein each Each substituent is independently, for example, halogen, amine, -(C3-C8)aryl, -(C3-C8)cycloalkyl, -O-(C3-C8)aryl, -O-(C3-C8)cycloalkyl, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; wherein the heteroatom in the heteroalkyl, heteroaryl or heterocycloalkyl is N, O or S. In a specific embodiment, X17 is any natural amino acid or any synthetic amino acid, the side chain of the natural amino acid or the synthetic amino acid is -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, substituted or unsubstituted. In one embodiment, X17 is Phe, cypTyrOBn, cypTyr, dcypTyrOBn, TyrOBn, B1, B2, B3 or B4.

[0207] In specific embodiments, the compound of formula (V) is any one of compounds 62-70 of Table II.

[0208] The present disclosure includes compounds of formula (V), wherein each of X1 to X17 is independently defined using any of the more general or more specific definitions provided above for these residues in formula (V).

[0209] In specific embodiments of Formula (V), at least 4 (or at least 5, 6, 7, 8, 9, 10, 11 or 12) of the residues at positions Xn defined above, wherein n is 1 to 12 and 14 to 17 (i.e., except for the closed loop residues X10 and X13, which are different from the corresponding residues in AP-17 (SEQ ID NO: 86)), correspond to residues in AP-17 (SEQ ID NO: 86). For example, compound 62 has 11 residues corresponding to residues in AP-17 (SEQ ID NO: 86). In other specific embodiments of Formula (V), except for the closed loop residues that are different from the corresponding residues in AP-13 (SEQ ID NO: 47), at least 4 (or at least 5, 6, 7, 8 or 9) of the residues correspond to residues in AP-13 (SEQ ID NO: 47). For example, compound 62 has 8 residues corresponding to residues in AP-13 (SEQ ID NO:47).

[0210] In other specific embodiments, the compounds of the present disclosure correspond to macrocyclic analogs of Ap17KFRRQRPRLSHKGPMPF (SEQ ID NO: 86), wherein the compounds differ from Ap17 by at least two substitutions and at least one (or 2, 3, 4, 5, 6, 7, 8, 9 or 10) additional substitutions, deletions and / or insertions at the positions of the closed loop. These substitutions, deletions and / or insertions are defined in the various Xn of formula (V) above. The correspondence between these Xn and Ap13 and A17 is shown in Table B below, where the "[" and "]" symbols are used to represent the ring closure residues in formula (V) and the positions in the compounds of the present disclosure that satisfy these formulas.

[0211] Elabela cyclic analogs

[0212] The present disclosure also encompasses Elabela cyclic analogs, such as those described in any one of Formulas (VI) to (VIII).

[0213] In one embodiment, the elabela cyclic analog comprises or consists of the following formula (VI): c[X1'-X2'-X3'-X4']c-X5'-X6'-c[C-X7'-X8'-X9'-C]c-X10'-X11'-X12'-X13'-X14',

[0214] in:

[0215] X1' and X4' are closed rings and are identical or different, and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, it is a residue carrying a terminal olefin or a free carboxyl or amine function. In a specific embodiment, it is each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α, γ-diamino-butyric acid, Nγ-allyl-α, γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α, γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, one of X1' and X4' is Glu, and the other is Lys. In a specific embodiment, it is Lys and Glu or Glu and Lys;

[0216] X2', X3', X9' and X10' are each independently a natural or synthetic amino acid, the side chain of which is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X2', X3', X9' and X10' are each independently absent or an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4. In a specific embodiment, X2', X3', X9' and X10' are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, His or absent. In a more specific embodiment, X2' and X3' are each independently -CH2-(CH2)p-guanidine, wherein p is 0 to 4; or Arg; and / or X9' is -(CH2)p-imidazole, wherein p is 0 to 4; or His; and / or X10' is -(CH2)p-(C3-C8)aryl, -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4; or Arg, Orn, Lys or 4-aminomethyl-phenylalanine;

[0217] X12' and X14' are each independently Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X12' and / or X14' are Pro;

[0218] X5', X7' and X11' are each independently any natural amino acid; or any synthetic amino acid, the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, The heterocycloalkyl, aryl or heteroaryl are optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amine, -OH, S or (C1-C6)alkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are N, O or S. In other specific embodiments, X5', X7' and X11' are Nle, Leu, Ala, Val, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine, α-methylphenylalanine, Trp, thiazol-5-yl-alanine, 3-(2-pyridyl)-alanine, 3-(3-pyridyl)-alanine or 3-(4-pyridyl)-alanine. In specific embodiments, X5', X7' and X11' are each independently -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)hydroxyalkyl, wherein p is 0 to 5. In another specific embodiment, X5', X7' and X11' are each independently Nle, Leu, Ala, Val, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine) or α-methylphenylalanine. In a more specific embodiment, X5' is Nle; and / or X7' is Leu; and X11' is Val; and

[0219] X6' and X8' are each independently absent or are each independently any natural amino acid; or any synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl , the heterocycloalkyl, the aryl or the heteroaryl are optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the alkyl, the heteroaryl, the aryl, the cycloalkyl and the heterocycloalkyl are optionally substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amine, -OH, S or (C1-C6)alkyl, and wherein the heteroatoms in the heteroalkyl, the heteroaryl or the heterocycloalkyl are N, O or S. In a more specific embodiment, X6' and / or X8' are absent.

[0220] X13' is an amino acid whose side chain is -(CH2)p-(C3-C8)aryl, wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently halogen, amine, -OH, S or (C1-C6)alkyl. In a specific embodiment, X14' is Phe or Phe substituted with halogen, such as bromophenyl.

[0221] In a specific embodiment, the compound of formula (VI) is compound 79 in Table III below.

[0222] The present disclosure includes compounds of formula (VI), wherein each of X1' to X14' is independently defined using any of the more general or more specific definitions provided above for these residues in formula (VI).

[0223] In another specific embodiment, the compound comprises or consists of the following formula (VII): X1'-X2'-X3'-X4'-c[X5'-X6'-X7'-X8'-X9'-X10']c-X11'-X12'-X13'-X14'-X15'-X16'-X17',

[0224] in:

[0225] X1' is absent or is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, a natural amino acid, a synthetic amino acid, the side chain of the natural amino acid, the synthetic amino acid is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -C(O)-(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally substituted with one or Two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl are fused; and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted by one or more substituents, wherein each substituent is independently, for example, halogen, amine, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, -O-(CH2)p-(C3-C8)aryl, wherein p is 0 to 3, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are N, O or S. In one embodiment, X1 is Pyr, -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11. In a more specific embodiment, X1 is Pyr.

[0226] X2', X3' and X13' are each independently a natural or synthetic amino acid, the side chain of which is -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X2', X3' and X13' are each independently an amino acid, the side chain of which is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine, wherein p is 0 to 4, optionally substituted by, for example, aryl. In a specific embodiment, X2', X3' and X13' are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg or His. In a more specific embodiment, X2', X3' and X13' are each independently -CH2-(CH2)p-guanidine, wherein p is 0 to 4, optionally substituted by, for example, aryl. In a more specific embodiment, X2' and X3' are each independently Arg, aryl-substituted Arg (e.g., -C(O)-(C3-C8)aryl, such as 4-bromobenzoyl), hArg, Nle, Leu, Ala, Val, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine) or α-methylphenylalanine. In a more specific embodiment, X2' and / or X3' are each independently Arg or hArg (substituted or unsubstituted (e.g., Arg substituted with 4-bromobenzoyl)); and X13' is Arg.

[0227] X4', X6', X8' and X12' are each independently absent or are any natural amino acid or synthetic amino acid whose side chain is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、-CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidino, -OH, S or (C1-C6)alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a more specific embodiment, X4', X6', X8' and X12' are each independently -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)hydroxyalkyl, wherein p is 0 to 5. In a more specific embodiment, X4', X6', X8' and X12' are each independently Leu, Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Ala, Val, Ile, Ser or Thr. In a more specific embodiment, X4', X6', X8' and X12' are each independently Ser, Nle or Leu. In a more specific embodiment, X4' and / or X12' are Ser; and / or X6' is Nle; and / or X8' is Leu.

[0228] X5' and X10' are ring-closed and identical or different and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, it is a residue carrying a terminal olefin or a free carboxyl or amine function. In a specific embodiment, it is each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α, γ-diamino-butyric acid, Nγ-allyl-α, γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α, γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, Lys, Orn, Dab, Dap, Asp, Glu or allyl Gly are present independently, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, one of X1' and X4' is Glu and the other is Lys. In a specific embodiment, it is Lys and Glu or Glu and Lys;

[0229] X7', X15' and X17' are each independently Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn). In a specific embodiment, X7', X15' and X17' are each Pro.

[0230] X9' and X11' are each independently absent or a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S, preferably N. In one embodiment, X9' and X11' are each independently absent or an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, wherein p is 0 to 4. In one embodiment, X9' is Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg, His or absent. In a more specific embodiment, X9' and X11' are each independently absent or -(CH2)p-imidazole, wherein p is 0 to 4, or His.

[0231] X14' is any natural amino acid; or any synthetic amino acid, the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C8)cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl groups; and wherein the alkyl, heteroaryl, aryl, cycloalkyl and heterocycloalkyl groups are optionally substituted with one or more substituents, wherein each substituent group is independently, for example, halogen, amine, -OH, S or (C1-C6)alkyl, and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl groups are N, O or S. In another specific embodiment, X14' is Nle, Leu, Ala, Ile, Val, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine) or α-methylphenylalanine. In specific embodiments, X14' is -(CH2)p-(C3-C8)alkyl, wherein p is 0 to 5, or is Ala, Val, Ile, Nle or Leu; and

[0232] X16' is an amino acid whose side chain is -(CH2)p-(C3-C8)aryl, wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently halogen, amine, -OH, S or (C1-C6)alkyl. In a specific embodiment, X14' is Phe or Phe substituted with halogen, such as bromophenyl.

[0233] In a specific embodiment, the compound of formula (VII) is any one of compounds 74-78 and 89-94 in Table III below. In another specific embodiment, the compound of formula (VII) is any one of compounds 77, 89, 91 and 94 in Table III below.

[0234] The present disclosure includes compounds of formula (VII), wherein each of X1' to X16' is independently defined using any of the more general or more specific definitions provided above for these residues in formula (VII). In another specific embodiment, the compound comprises or consists of the following formula (VIII): c[X1'-X2'-X3'-X4']c-X5'-X6'-X7'-X8'-X9'-X10'-X11'-X12'-X13'-X14'

[0235] in:

[0236] X1' and X4' are ring-closed and identical or different and are aliphatic residues, alkenyl residues, acid residues or natural or non-natural amino acids or derivatives thereof, which are optionally substituted. In a specific embodiment, they are residues carrying terminal olefins or free carboxyl or amine functions. In a specific embodiment, they are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Asp, Glu, allyl Gly, Nα-allyl-arginine, Nπ-allyl-histidine, Nγ-allyl-Nγ-nitrobenzenesulfonyl-α, γ-diamino-butyric acid, Nγ-allyl-α, γ-diamino-butyric acid or Nγ-allyl-Nγ-methyl-α, γ-diamino-butyric acid, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, Lys, Orn, Dab, Dap, Asp, Glu or allyl Gly are present independently, wherein the cycle is closed by an amide bridge or an olefin. In a specific embodiment, one of X1' and X4' is Glu and the other is Lys. In a specific embodiment, it is Lys and Glu or Glu and Lys;

[0237] X2', X3', X8' and X10' are each independently a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH2, -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl The alkyl, heterocycloalkyl, aryl or heteroaryl are optionally substituted with at least one amino or guanidino group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl are optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl groups; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl groups are 1, 2 or 3 N, O or S, preferably N. In a specific embodiment, X2', X3', X8' and X10' are each independently an amino acid, the side chain of which is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, wherein p is 0 to 4. In a specific embodiment, X2', X3', X8' and X10' are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg or His. In a more specific embodiment, X2', X3', X8' and X10' are each independently -CH2-(CH2)p-guanidine or -(CH2)p-imidazole, wherein p is 0 to 4. In another specific embodiment, X2', X3' and X10' are each independently -CH2-(CH2)p-guanidine, wherein p is 0 to 4. In another specific embodiment, X2', X3' and X10' are each independently Arg or hArg; and / or X8' is His.

[0238] X5', X7', X9' and X11' are each independently any natural amino acid; or a synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH2, -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidino, -OH, S or (C1-C6) alkyl; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8) aryl, (C3-C8) heteroaryl, (C3-C8) cycloalkyl or -(C3-C8) heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In another specific embodiment, X5', X7', X9' and X11' are each independently -(CH2)p-(C3-C8) alkyl, wherein p is 0 to 5. In another specific embodiment, X5', X7', X9' and X11' are each independently Leu, Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Ala, Val, Ile, Ser or Thr. In a more specific embodiment, X5', X7', X9' and X11' are each independently Ser, Nle, Leu or Val. In a more specific embodiment, X5' is Nle; and / or X7' is Leu; and / or X9' is Ser; and / or X11' is Val.

[0239] X6', X12' and X14' are each independently Gly, Phe, Leu, Ile, Ser, Pro, Aib, Sar, Oic, βAla, Hyp or Hyp(OBn), preferably Pro or Oic. In a specific embodiment, X6', X12' and X14' are each Pro.

[0240] X13' is an amino acid whose side chain is -(CH2)p-(C3-C8)aryl, wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently halogen, amine, -OH, S or -(C1-C6)alkyl. In another specific embodiment, the X18 is Phe or Phe substituted with halogen. In another embodiment, the X18 is Phe.

[0241] In a specific embodiment, the compound of formula (VIII) is any one of compounds 72 and 73 in Table III below. In a more specific embodiment, the compound of formula (VIII) is compound 72.

[0242] In specific embodiments of formula (VI) to (VIII), at least 4 (or at least 5, 6, 7, 8, 9, 10, 11 or 12) of the residues (e.g., at positions Xn defined above, where n is 1 to 14 in formula (VI) and (VIII), or 1 to 17 in formula (VII)) correspond to the residues in ELA (19-32) (SEQ ID NO: 85), except for the residues of the closed loop that are different from the corresponding residues in ELA (19-32) (SEQ ID NO: 85). For example, compound 79 has 8 residues corresponding to the residues in ELA (19-32) (SEQ ID NO: 85).

[0243] In other specific embodiments, the compounds of the present disclosure correspond to macrocyclic analogs of Ela (19-32) PyrRRCMPLHSRVPFP (SEQ ID NO: 85), wherein the compounds differ from Ela (19-32) in at least two substitutions and at least one (or 2, 3, 4, 5, 6, 7 or 8) additional substitutions, deletions and / or insertions at the position of the closed loop. These substitutions, deletions and / or insertions are defined in the various Xn of formulas (VI) to (VIII) above. The correspondence between these Xn and ELA (19-32) is shown in Table C below, where the "[" and "]" symbols are used to represent the ring closure residues in formulas (VI) to (VIII) and the positions of the compounds of the present disclosure that satisfy these formulas.

[0244] In formulae (I) to (VIII), unless otherwise stated, the link between two amino acid residues (natural or non-natural) is a peptide bond.

[0245] In another specific embodiment of formula (I) to (VIII), one of the ring-closing residues is Lys, Dap, Dab, Orn, and the other is Glu or Asp.

[0246] In one embodiment, one of the terminal (natural or non-natural) amino acid residues used for closure is a (natural or non-natural) amino acid with an amine on its side chain before ring closure, and the other is a (natural or non-natural) amino acid with a carboxylic acid on its side chain before ring closure, so that the amine and the carboxylic acid form an amide by macrolactamization. More specifically, one of the terminal (natural or non-natural) amino acid residues can be substituted before ring closure. After closure, the side chain carboxylic acid activated by the coupling agent has reacted with the amino group of the side chain of the other residue to form a peptide bond using, for example, a macrolactamization reaction.

[0247] As used herein, the term "substituted" with reference to the above-listed natural or non-natural amino acids or acid residues in the structures refers to substitution with halogen (e.g., Cl, F, Br, I), -OH, (C1-C6)alkyl, hydroxy(C1-C6)alkyl, (C3-C6)aryl, (C3-C6)aryl(C1-C6)alkyl, (C3-C6)cycloalkyl, hetero(C3-C6)aryl, hetero(C3-C6)alkyl, C6)aryl(C1-C6)alkyl, hetero(C3-C6)cyclo(C1-C6)alkyl, amino(C1-C6)alkyl, amino(C3-C6)aryl, amino(C3-C6)aryl(C1-C6)alkyl, amino(C3-C6)cycloalkyl, aminohetero(C3-C6)aryl, aminohetero(C3-C6)aryl(C1-C6)alkyl or aminohetero(C3-C6)cyclo(C1-C6)alkyl.

[0248] In specific embodiments, the size of the macrocyclic ring can be 14 to 24 ring atoms (or 15 to 23, 16 to 22, 17 to 20). In specific embodiments, the size of the macrocyclic ring can be 17 to 20 ring atoms.

[0249] In all of the aforementioned compounds, the residues (e.g., X1 to Xn) can be in the L or D configuration. In all of the aforementioned combinations of two residues, they can be in the L, L; LD; D, L; or D, D configuration.

[0250] Without being limited thereto, specific compounds of the present disclosure encompass those compounds satisfying Formulae (I) to (VIII). In a more specific embodiment, the compounds of the present disclosure encompass the compounds listed in Tables I to III.

[0251] Chemical Group

[0252] As used herein, the term "alkyl" refers to a monovalent straight or branched, saturated or unsaturated aliphatic hydrocarbon group having a number of carbon atoms within a specified range. Thus, for example, "(C1-12) alkyl" (or "C1-12 alkyl") refers to any alkyl group having up to 12 carbon atoms, including hexylalkyl and pentylalkyl isomers, as well as n-butyl, isobutyl, sec-butyl and tert-butyl, n-propyl and isopropyl, ethyl and methyl. As another example, "(C1-4) alkyl" refers to n-butyl, isobutyl, sec-butyl and tert-butyl, n-propyl and isopropyl, ethyl and methyl. As another example, "C1-3 alkyl" refers to n-propyl, isopropyl, ethyl and methyl. Alkyl includes unsaturated aliphatic hydrocarbons, including alkynes (RC≡CR); and / or alkenes (RC=CR).

[0253] The term "halogen" (or "halo") refers to fluorine, chlorine, bromine and iodine (alternatively referred to as fluoro, chloro, bromo and iodo). The term "haloalkyl" refers to an alkyl group as defined above in which one or more of the hydrogen atoms has been replaced by a halogen (i.e., F, Cl, Br and / or I). Thus, for example, "C1-10 haloalkyl" (or "C1-C6 haloalkyl") refers to a C1 to C10 straight or branched chain alkyl group as defined above having one or more halogen substituents. The term "fluoroalkyl" has a similar meaning, except that the halogen substituents are limited to fluorine. Suitable fluoroalkyl groups include (CH2) 0-4 CF3 series (ie, trifluoromethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluoro-n-propyl, etc.).

[0254] The term "heteroalkyl" is given its ordinary meaning in the art and refers to an alkyl group as described herein in which one or more carbon atoms are replaced by a heteroatom (e.g., oxygen, nitrogen, sulfur or derivatives thereof, etc.). Examples of heteroalkyl groups include, but are not limited to, alkoxy groups, alkyl-substituted amino groups, thiols, such as methionine side groups. Up to two heteroatoms may be consecutive. When a prefix such as C 2-6 When used to refer to heteroalkyl groups, the number of carbons (2-6 in this example) is meant to include heteroatoms as well.

[0255] The term "aminoalkyl" refers to an alkyl group as defined above in which one or more of the hydrogen or carbon atoms have been replaced by nitrogen or an amino derivative such as, but not limited to, guanidine. Thus, for example, "C 1-6 "Aminoalkyl" (or "C1-C6 aminoalkyl") refers to a C1 to C6 straight or branched chain alkyl group as defined above having one or more amino derivatives (e.g., NH, amide, diazine, azide, etc.).

[0256] The term "thioalkyl" refers to an alkyl group as defined above in which one or more of the hydrogen or carbon atoms has been replaced by a sulfur atom or a thiol derivative. Thus, for example, "C 1-6 "Thioalkyl" (or "C1-C6 thioalkyl") refers to a C1 to C6 straight or branched chain alkyl group as defined above having one or more sulfur atoms or thiol derivatives (eg, S, SH, etc.).

[0257] Aminoalkyl and thioalkyl are specific examples of and are encompassed by the term "heteroalkyl" or substituted alkyl, depending on the heteroatom replacing a carbon or hydrogen atom.

[0258] The term "cycloalkyl" refers to a saturated alicyclic hydrocarbon consisting of a saturated 3-8 membered ring, which is optionally fused to an additional (1-3) aliphatic (cycloalkyl) or aromatic ring system, each additional ring consisting of a 3-8 membered ring. It includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl (cyp) (e.g., compounds 38-41 and 63-65), cyclohexyl and cycloheptane.

[0259] The term "heterocyclyl" refers to: (i) a 4- to 7-membered saturated heterocyclic ring containing 1 to 3 heteroatoms independently selected from N, O and S; or (ii) a heterobicyclic ring (e.g., benzocyclopentyl, octahydroindole (e.g., compound 166)). Examples of 4- to 7-membered saturated heterocyclic rings within the scope of the present disclosure include, for example, azetidinyl, piperidinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, isothiazolidinyl, oxazolidinyl, isoxazolidinyl, pyrrolidinyl, pyridine, imidazolidinyl, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, pyrazolidinyl, hexahydropyrimidinyl, thiazinanyl, thiazepanyl, azepanyl, diazepanyl, tetrahydropyranyl, tetrahydrothiopyranyl and dioxanyl. Examples of 4- to 7-membered unsaturated heterocycles within the scope of the present disclosure include monounsaturated heterocycles corresponding to the saturated heterocycles listed in the preceding sentence, wherein a single bond is replaced by a double bond (eg, a carbon-carbon single bond is replaced by a carbon-carbon double bond).

[0260] The term "C(O)" refers to a carbonyl group. The terms "S(O)2" and "SO2" each refer to a sulfonyl group. The term "S(O)" refers to a sulfinyl group.

[0261] The term "aryl" refers to an aromatic (unsaturated) compound consisting of 3-8 ring members, optionally fused to an additional (1-3) aliphatic (cycloalkyl) or aromatic ring system, each additional ring consisting of 3-8 ring members (such as anthracene, indane, Tic, 3-benzothienylalanine or dihydroindole). In a specific embodiment, it refers to phenyl, benzocyclopentyl or naphthyl.

[0262] The term "heteroaryl" refers to (i) a 3-, 4-, 5-, 6-, 7- or 8-membered heteroaromatic ring (more specifically a 3-7- or 3-6-membered ring) containing 1 to 4 heteroatoms independently selected from N, O and S, such as thiophenyl, thienyl, pyridine, or (ii) a heterobicyclic ring selected from indolyl, quinolyl, isoquinolyl, Tic, indolinylglycine and quinoxalinyl. Suitable 3-, 4-, 5- and 6-membered heteroaromatic rings include, for example, diazirine, pyridyl (also referred to as pyridinyl), pyrrolyl, diazines (e.g., pyrazinyl, pyrimidinyl, pyridazinyl), triazinyl, thienyl, furanyl, imidazolyl, pyrazolyl, triazolyl (e.g., 1,2,3 triazolyl), tetrazolyl (e.g., 1,2,3,4 tetrazolyl), oxazolyl, isoxazolyl, oxadiazolyl, oxatriazolyl, thiazolyl, isothiazolyl and thiadiazolyl. Of particular interest are heteroaromatics pyrrolyl, imidazolyl, pyridyl, pyrazinyl, quinolinyl (or quinolyl), isoquinolinyl (or isoquinolyl) and quinoxalinyl. Suitable heterobicyclic rings include indolyl.

[0263] The term "aralkyl" and more specifically "(C4-C14)aralkyl" or "C4-14aralkyl" herein refers to compounds comprising 3-7 ring-membered aryl groups substituted with 1 to 7 alkyl groups. In specific embodiments, it refers to benzyl or ethoxyphenyl.

[0264] As used herein, and unless otherwise indicated, the terms "alkyl", "haloalkyl", "aminoalkyl", "cycloalkyl", "heterocyclyl", "aryl", "heteroalkyl" and "heteroaryl" and terms representing specific embodiments thereof (e.g., butyl, fluoropropyl, aminobutyl, cyclopropane, morpholine, phenyl, pyrazole, etc.) encompass substituted embodiments (i.e., in the case of haloalkyl and aminoalkyl, except for their halogen and nitrogen substituents, respectively) and unsubstituted embodiments of these groups. Thus, for example, the term "phenyl" encompasses unsubstituted phenyl as well as fluorophenyl, hydroxyphenyl, methylsulfonylphenyl (or biphenyl), diphenyl, trifluoromethyl-diazine-phenyl, isopropyl-phenyl, trifluorohydroxy-phenyl. Similarly, the term pyrazole encompasses unsubstituted pyrazole as well as methylpyrazole. One or more substituents may be amine, halogen, hydroxy, C1-6 aminoalkyl, C1-6 heteroalkyl, C1-6 alkyl, C3-8 cycloalkyl, C1-6 haloalkyl, aryl, heteroaryl and heterocyclyl (etc.).

[0265] It should be understood that the specific rings listed above are not limiting of the rings that can be used in the present disclosure. These rings are merely representative.

[0266] Unless clearly stated to the contrary in a specific context, any of the various rings and ring systems described herein may be attached to the rest of the compound at any ring atom (ie, any carbon atom or any heteroatom) from which a stable compound is produced.

[0267] Isomers, tautomers and polymorphs

[0268] As used herein, the term "isomer" refers to stereoisomers, including optical isomers (enantiomers), diastereomers, and other known types of isomers.

[0269] The compounds of the present disclosure have at least 5 asymmetric carbon atoms and therefore can exist in the form of optically pure enantiomers (optical isomers) and as mixtures thereof (racemates). It should be understood that, unless otherwise indicated, the present disclosure includes racemates, enantiomers and / or diastereomers of the compounds of the present disclosure and mixtures thereof. In addition, some macrocyclic compounds of the present invention include closed-cycle olefins. Such compounds have Z and E isomers.

[0270] For further clarity, (S)-H or (S)-CH3 indicates that a stereogenic center having an H or CH3 substituent has a (S) stereochemistry.

[0271] In addition, the present disclosure includes all geometric isomers. For example, when the compounds of the present disclosure contain double bonds or fused rings, both cis-forms and trans-forms and mixtures are included within the scope of the present disclosure.

[0272] In the present disclosure, it should be understood that the compounds of the present disclosure can exhibit tautomerism, and the formula diagrams in this specification may represent only one of the possible tautomeric forms. It should be understood that the present disclosure encompasses any tautomeric form, and is not limited to any one tautomeric form used in the formula diagram.

[0273] It is also to be understood that certain compounds of the disclosure may exhibit polymorphism and that the disclosure encompasses all such forms.

[0274] Salt

[0275] The present disclosure relates to compounds of the present disclosure as defined above and salts thereof. As used herein, the term "salt" means a basic salt formed from an inorganic base and / or an organic base. Salts used in pharmaceutical compositions will be pharmaceutically acceptable salts, but other salts may also be used in the production of compounds of the present disclosure. The term "pharmaceutically acceptable salt" refers to salts of compounds of the present disclosure that are pharmaceutically acceptable and substantially nontoxic to the subject to which they are administered. More specifically, these salts retain the biological effectiveness and properties of the anti-atherosclerotic compounds of the present disclosure and are formed from suitable nontoxic organic or inorganic acids or bases.

[0276] For example, in the case where the compounds of the present disclosure are sufficiently acidic, the salts of the present disclosure include basic salts formed from inorganic bases or organic bases. Such salts include alkali metal salts such as sodium salts, lithium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; metal salts such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts and cobalt salts; inorganic amine salts such as ammonium salts or substituted ammonium salts, for example trimethylammonium salts; and salts with organic bases (e.g., organic amines), such as chloroprocaine salts, dibenzylamine salts, dicyclohexylamine salts, dicyclohexylamine, diethanolamine salts, ethylamine salts (including diethylamine salts and trimethylamine salts). The preferred salts include methylamine salts (including dimethylamine salts and trimethylamine salts), morpholine salts, morpholine salts, N,N'-dibenzylethylenediamine salts, N-benzyl-phenylethylamine salts, N-methylglucosamine salts, phenylglycine alkyl ester salts, piperazine salts, piperidine salts, procaine salts, tert-butylamine salts, tetramethylammonium salts, tert-octylamine salts, tris-(2-hydroxyethyl)amine salts and tris(hydroxymethyl)aminomethane salts. Preferred salts include salts formed from sodium, lithium, potassium, calcium and magnesium.

[0277] Such salts can be routinely formed by those skilled in the art using standard techniques. In fact, chemical modification of pharmaceutical compounds (i.e., drugs) into salts is a technique known to pharmaceutical chemists (see, for example, H.Ansel et al., " Pharmaceutical Dosage Forms and Drug Delivery Systems (Pharmaceutical Dosage Forms and Drug Delivery Systems)" (6th edition 1995) page 196 and page 1456-1457, incorporated herein by reference). The salt of a compound of the present disclosure can be formed by, for example, reacting a compound of the present disclosure with a certain amount of acid or alkali (such as an equivalent amount of acid or alkali) in a medium (a medium of salt precipitation or in an aqueous medium) and then lyophilizing it.

[0278] ester

[0279] The present disclosure relates to compounds of the present disclosure as defined above and esters thereof. As used herein, the term "ester" refers to a compound of the present disclosure or a salt thereof in which the carboxylic acid is a hydroxyl group and has been converted to the corresponding ester using an alcohol and a coupling reagent. Esters used in pharmaceutical compositions will be pharmaceutically acceptable esters, but other esters may also be used in the production of compounds of the present disclosure.

[0280] The term "pharmaceutically acceptable ester" refers to esters of the compounds of the present disclosure that are pharmaceutically acceptable and substantially non-toxic to the subject to which they are administered. More specifically, these esters retain the biological effectiveness and properties of the anti-atherosclerotic compounds of the present disclosure and act as prodrugs that, when absorbed into the bloodstream of warm-blooded animals, are cleaved in a manner that yields the parent alcohol compound.

[0281] Esters of the compounds of the present disclosure particularly include the following groups: (1) carboxylic acid esters obtained by esterification, wherein the non-carbonyl portion of the carboxylic acid portion of the ester group is selected from linear or branched alkyl (e.g., ethyl, n-propyl, tert-butyl, n-butyl, methyl, propyl, isopropyl, butyl, isobutyl or pentyl), n-hexyl, alkoxyalkyl (e.g., methoxymethyl, acetoxymethyl and 2,2-dimethylpropionyloxymethyl), aralkyl (e.g., benzyl), aryloxyalkyl (e.g., phenoxymethyl), aryl (e.g., phenyl optionally substituted by, for example, halogen, C1-4 alkyl, or C1-4 alkoxy or amino).

[0282] More information on examples and uses of esters for delivering drug compounds can be found in Design of Prodrugs. Bundgaard H, ed. (Elsevier, 1985), which is incorporated herein by reference. See also H. Ansel et al., 1995, pp. 108–109; Krogsgaard-Larsen, 1996, pp. 152–191; Jarkko Rautio, 2008; and Pen-Wei Hsieh, 2009, all of which are incorporated herein by reference.

[0283] The compounds of the present disclosure can be esterified by a variety of conventional procedures involving the formation of esters from the acid of the molecule by reaction with a coupling agent such as DIC (diisopropylcarbodiimide) and a base such as NN-dimethylaminopyridine (DMAP) and an alcohol such as methanol (methyl ester), ethanol, a long chain alcohol or benzyl alcohol (benzyl ester). Those skilled in the art will readily appreciate how to successfully carry out these and other known acid esterification methods.

[0284] Esters of the compounds of the present disclosure may form salts. In this case, this is achieved by conventional techniques as described above.

[0285] Solvate

[0286] The compounds of the present disclosure may exist in unsolvated as well as solvated forms with solvents such as water, ethanol, and the like, and it is intended that the present disclosure includes both solvated and unsolvated forms.

[0287] "Solvate" means a physical association of a compound of the present disclosure with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, the solvate will be capable of separation, for example when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" encompasses both solution phase and isolatable solvates. Solvates used in pharmaceutical compositions will be pharmaceutically acceptable solvates, but other solvates may also be used in the production of compounds of the present disclosure.

[0288] As used herein, the term "pharmaceutically acceptable solvate" means a solvate of the compounds of the present disclosure that is pharmaceutically acceptable and substantially non-toxic to the subject to which it is administered. More specifically, these solvates retain the biological effectiveness and properties of the anti-atherosclerotic compounds of the present disclosure and are formed from suitable non-toxic solvents.

[0289] Non-limiting examples of suitable solvates include glycolates, methanolates, and the like, as well as hydrates, which are solvates wherein the solvent molecule is H2O.

[0290] The preparation of solvates is generally known. Thus, for example, Caira, 2004 (incorporated herein by reference) describes the solvate of the antifungal fluconazole in ethyl acetate and the preparation of solvates from water. van Tonder, 2004; Bingham, 2001 (both of which are incorporated herein by reference) describe similar preparations of solvates, hemisolvates, hydrates, etc.

[0291] A typical non-limiting method for preparing a solvate involves dissolving a compound of the invention in a desired amount of a desired solvent (organic or aqueous or a mixture thereof) at a temperature above ambient temperature and cooling the solution at a rate sufficient to form crystals which are then separated by standard methods. Analytical techniques such as IR spectroscopy can be used to show the presence of a solvent (or water) in the crystals as a solvate (or hydrate).

[0292] Compositions, kits and combinations

[0293] Composition

[0294] The present disclosure also relates to a pharmaceutical composition comprising the above-mentioned compound of the present disclosure or a pharmaceutically acceptable salt, ester and solvate thereof and optionally a pharmaceutically acceptable carrier.

[0295] As used herein, the term "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and generally do not produce allergic or similar adverse reactions (such as stomach discomfort, dizziness, etc.) when administered to a subject (e.g., a human). Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0296] The term "carrier" refers to a diluent, adjuvant, excipient or vehicle that can be administered with the compounds of the present disclosure. Sterile water or aqueous saline solutions and aqueous dextrose and glycerol solutions can be used as carriers, especially for injectable solutions. Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin. The pharmaceutical compositions of the present disclosure may also contain excipients / carriers, such as preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, odorants, salts for osmotic pressure changes, buffers, coating agents or antioxidants.

[0297] In some embodiments, the compositions provided herein are administered by one or more administration routes using one or more of a variety of suitable methods. As will be appreciated by those skilled in the art, the administration route and / or mode will vary according to the desired result. The compounds disclosed herein are used for administration routes for purposes disclosed herein, including but not limited to intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral administration routes, such as by injection or infusion. As used herein, the phrase "parenteral administration" means an administration mode, usually by injection, in addition to enteral administration and topical administration, and includes but is not limited to intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion. Alternatively, the compounds of the disclosure provided herein are administered by a non-parenteral route, such as oral (see, e.g., US 7,875,648 B2 to Meier), topical, epidermal or mucosal administration routes, e.g., intranasal, oral, vaginal, rectal, sublingual or topical administration.

[0298] Without limitation, when the compounds / pharmaceutical compositions of the present disclosure are administered orally, they can be in the form of, for example, tablets, coated tablets, dragees, hard or soft gelatin capsules, solutions, emulsions or suspensions; rectal administration, for example, using suppositories; local, topical or transdermal administration, for example, using ointments, creams, gels or solutions; or parenteral administration, for example, intravenous, intramuscular, subcutaneous, intrathecal or transdermal administration, using, for example, injectable solutions. In addition, administration can be performed sublingually, nasally, or as an ophthalmic preparation or aerosol, for example, in the form of a spray such as a nasal spray.

[0299] The compounds of the present disclosure can be combined with any vehicle commonly used in pharmaceutical preparations into dosage forms. Methods for preparing appropriate formulations are well known in the art (see, for example, Remington's Pharmaceutical Sciences, 16th edition, 1980, edited by A. Oslo, Easton, Pennsylvania, the document being incorporated herein by reference). Common pharmaceutically acceptable carriers include, but are not limited to, sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters. Aqueous carriers include, but are not limited to, water, alcohol, saline, and buffered solutions. Pharmaceutically acceptable carriers may also include physiologically acceptable aqueous vehicles (e.g., normal saline) or other known carriers suitable for specific routes of administration.

[0300] In order to prepare tablets, coated tablets, dragees or hard gelatin capsules, the compounds of the present disclosure can be mixed with any known pharmaceutically inert inorganic or organic excipients and / or carriers. Examples of suitable excipients / carriers include lactose, corn starch or derivatives thereof, talcum or stearic acid or salts thereof. Suitable excipients used together with soft gelatin capsules include, for example, vegetable oils, waxes, fats, semisolid or liquid polyols, etc. However, according to the properties of the active ingredient, soft gelatin capsules may not require excipients at all. For the preparation of solutions and syrups, usable excipients include, for example, water, polyols, sucrose, invert sugar and glucose.

[0301] For suppositories and topical or percutaneous administration, excipients which may be used include, for example, natural or hardened oils, waxes, fats and semi-solid or liquid polyols.

[0302] In the case of selecting parenteral administration as the route of administration, the preparation containing the compound of the present disclosure can be provided to the patient in combination with a pharmaceutically acceptable sterile aqueous or non-aqueous solvent, suspension or emulsion. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oil, fish oil and injectable organic esters. Aqueous carriers include water, water-alcohol solutions, emulsions or suspensions, including saline and buffered medical parenteral vehicles, including sodium chloride solution, Ringer's dextrose solution (Ringer's dextrose solution), dextrose plus sodium chloride solution, Ringer's solution (Ringer's solution) containing lactose or fixed oil. Intravenous vehicles can include fluid and nutritional supplements, electrolyte supplements, such as those based on Ringer's dextrose, etc.

[0303] The medicament / pharmaceutical composition may also contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavoring agents, salts for variations in osmotic pressure, buffers, coating agents or antioxidants. It may also contain other therapeutically active agents.

[0304] In some embodiments, the active compound can be prepared with a carrier that will protect the compound from rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. In some embodiments, biodegradable polymers, biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid are used. Many methods for preparing such preparations are patented or generally known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, edited by JR Robinson, Marcel Dekker, Inc., New York, 1978. In some embodiments, the therapeutic composition is administered together with a medical device known in the art. For example, in one embodiment, the therapeutic composition provided herein is administered together with a needleless subcutaneous injection device.

[0305] Any amount of pharmaceutical composition can be administered to a subject. The dosage will depend on many factors, including the age and requirements of the patient and the mode of application. Generally, the amount of the compound of the present disclosure contained in a single dose will be an amount that effectively prevents, delays or treats the disease or condition to be treated without causing significant toxicity. Therefore, the "therapeutically effective amount" or "effective amount" or "therapeutically effective dose" of a specific compound of the present disclosure or its composition can reduce pain and / or lower body temperature in the subject. Intravenous or oral administration is a preferred form of use.

[0306] The effective amount of the compound of the present disclosure can also be directly measured. The effective amount can be given every day or every week or part thereof. Generally, the pharmaceutical composition of the present disclosure can be applied with an amount of about 0.001 mg to about 500 mg (e.g., 10 mg, 50 mg, 100 mg or 250 mg) per kg body weight every day. The dosage can be provided in a single dose regimen or a multiple dose regimen. For example, in some embodiments, the effective amount can be about 1 mg to about 25 grams of compositions every day, about 50 mg to about 10 grams of compositions every day, about 100 mg to about 5 grams of compositions every day, about 1 gram of compositions every day, about 1 mg to about 25 grams of compositions every week, about 50 mg to about 10 grams of compositions every week, about 100 mg to about 5 grams of compositions every other day, and about 1 gram of compositions once a week.

[0307] These are merely guidelines, as the actual dosage must be carefully selected and titrated by the attending physician based on clinical factors specific to each patient. The optimal daily dosage will be determined by methods known in the art and will be influenced by factors such as the patient's age and other clinically relevant factors. In addition, the patient may be taking medications to treat other diseases or conditions. Other medications may be continued during administration of the pharmaceutical compositions of the present disclosure to the patient, but in such cases it is particularly advisable to start with a low dose to determine whether adverse side effects are experienced.

[0308] combination

[0309] According to another aspect, there is provided a combination of at least one of the compounds described herein with another one of the compounds described herein and / or another drug.

[0310] Reagent test kit

[0311] According to another aspect of the present disclosure, a kit is provided, which comprises the compound defined herein or the above composition, and instructions for use thereof in preventing or treating cardiovascular diseases.

[0312] In a specific embodiment of a kit, the kit comprises: (i) at least one of the compounds described herein; (ii) another drug for preventing or treating cardiovascular disease; (iii) instructions for its use in preventing or treating cardiovascular disease; or (iv) a combination of at least two of (i) to (iii).

[0313] method

[0314] The present disclosure also relates to a method of preventing or treating cardiovascular disease or a symptom thereof in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of formula (I) and (II).

[0315] As used herein, the term "cardiovascular disease" refers to, but is not limited to, heart failure, pulmonary hypertension, cardiac dysfunction in sepsis, cardiac ischemia, and cerebral ischemia.

[0316] As used herein, the term "subject" refers to an animal, such as, but not limited to, a human or a pet or other animal (eg, pets such as cats, dogs, horses; as well as cattle, fish, pigs, poultry, etc.).

[0317] As used herein, the term "subject in need thereof" refers to a subject who will benefit from receiving an effective amount of a compound or composition of the present disclosure. In the context of a method for preventing or treating pain, a subject in need thereof refers to a subject experiencing or at risk of experiencing cardiovascular disease.

[0318] The use of the terms "a," "an," "the" and similar referents in the context of describing the disclosure (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0319] The terms "comprising," "having," "including," and "containing" should be construed as open-ended terms (ie, meaning "including, but not limited to,") unless otherwise noted.

[0320] Unless otherwise indicated herein, reference to ranges of values ​​herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, and each separate value is incorporated into the specification as if individually recited herein. All subsets of values ​​within the range are also incorporated into the specification as if they were individually recited herein.

[0321] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context.

[0322] The use of any and all examples, or exemplary language (eg, "such as") provided herein, is intended merely to better illuminate the disclosure and does not pose a limitation on the scope of the disclosure unless otherwise claimed.

[0323] No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0324] As used herein, the term "about" has its ordinary meaning. In embodiments, it may mean plus or minus 10% of a qualified value.

[0325] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0326] Other objects, advantages and features of the present disclosure will become more apparent from a reading of the following non-limiting description of specific embodiments of the present disclosure, which is given by way of example only with reference to the accompanying drawings.

[0327] The present disclosure is further illustrated by the following non-limiting examples.

[0328] Example 1 :Materials and methods

[0329] Reagents and instruments

[0330] All amino acids, HATU, DEPBT, DIPEA, piperidine, DIAD, and triphenylphosphine were purchased from Chem-Impex (Illinois, USA). Wang resin (capacity 0.4-0.7 mmol / g) was ordered from Rapp-polymer (Tuebingen, Germany). Hoveyda-Grubbs catalyst generation 2 was obtained from Sigma-Aldrich (Ontario, Canada). Other solvents and reagents were purchased from Fischer Scientific (Ontario, Canada) and used as received.

[0331] Thin layer chromatography (TLC) was performed on glass plates pre-coated with silica gel 60F254 (Merck, Darmstadt, Germany) and visualized with UV light (254 nm) and KMnO4 spray. Purification of organic molecules was performed by flash chromatography using a Biotage Isolera One system (Charlotte, North Carolina, US). High-resolution electrospray mass spectrometry (HRMS) data were recorded using a maXis ESI-Q-Tof instrument (Billerica, USA). A UPLC-MS system (Acquity ELISA, 1000 x 400,000) filled with 1.7 μm particles was used from Waters (Milford, USA). CSH TMAnalytical LC was performed on a C18 column (2.1 x 50 mm). Recordings were made on a Bruker Ascend 400 at 400 MHz and 100 MHz or on a Bruker 600 MHz Varian INOVA spectrometer at 600 MHz. 1 H and 13 C NMR spectrum (298K). Chemical shifts are expressed in parts per million (ppm). The residual solvent signal was used as an internal standard.

[0332] Fmoc-based amino acid synthesis

[0333] N α -Fmoc-(N π -allyl)-L-histidine-OH.

[0334] N α -(((9H-fluoren-9-yl)methoxy)carbonyl)-N τ -trityl-L-histidine methyl ester (Fmoc-L-His(Trt)-OMe 103). To a suspension of Fmoc-L-His(Trt)-OH (1 eq., 3.1 g, 5 mmol) and 1-hydroxy-benzotriazole monohydrate (HOBt HO, 1.1 eq., 1.05 g, 5.5 mmol) in 30 mL of DCM was added 1-ethyl-2-diaminopropyl-carbodiimide hydrochloride (EDC HCl, 1.1 eq., 743 mg, 5.5 mmol). A small amount of MgSO4 (about 100 mg) was added to absorb water, and the mixture was stirred at room temperature for 10 minutes. Methanol (1 mL) was added, and the reaction was run at room temperature overnight. The organic phase was washed twice with water, once with brine, dried over MgSO4 and filtered. The organic solvent was removed under vacuum, and the product was purified by flash chromatography using DCM-MeOH (95:5) as eluent. The product was obtained as a white solid, 2.33 g, 73% yield. The characterization of the product was the same as reported (Ahn et al., 2013).

[0335] N α -(((9H-fluoren-9-yl)methoxy)carbonyl)-N π -allyl-L-histidine methyl ester (N α -Fmoc-(N π-allyl)-L-histidine-OMe 104). A mixture of allyl alcohol (1.1 equivalents, 278 μL, 4.0mmol) and diisopropylethylamine (DIPEA, 1.2 equivalents, 855 μL, 4.4mmol) in 18mL DCM was added to a solution of freshly distilled Tf2O (1.1 equivalents, 686 μL, 4.0mmol) in 26mL DCM precooled at -78°C (dropwise addition for 15 minutes). The reaction mixture was stirred at -78°C for 30 minutes and slowly transferred to a solution of Fmoc-His (Trt)-OMe (1 equivalent, 2.32g, 3.7mmol) dissolved in 30mL DCM precooled at -78°C. After 10 minutes, the reaction mixture was warmed to room temperature and stirred at room temperature overnight. The residual acid was neutralized by vigorous mixing with 10mL saturated NaHCO3. The organic phase is washed twice with saturated NaHCO, dried with MgSO, filtered and evaporated to dryness. The trityl group is cut by treating the crude product with a solution of TFA / TIPS (2mL / 0.5mL) in 20mL DCM for 2 hours at room temperature. The mixture is evaporated to dryness, the residual acid is neutralized with 40mL saturated NaHCO, and the product is extracted with EtOAc. Purification is carried out using flash chromatography with DCM-MeOH (gradient 0 → 10% MeOH during 10CV). The product, 1.1g, yield 70% is obtained as white foam. 1 H NMR (400 MHz, CDCl3) δ (ppm) 8.58 (s, 1H), 7.74 (d, J = 7.5 Hz, 2H), 7.55 (d, J = 7.3 Hz, 2H), 7.38 (t, J = 7.4 Hz, 2H), 7.29 (t, J = 7.4 Hz, 2H), 7.18 (s, 1H), 6.09 (d, J = 7.2 Hz, 1H, proton amide), 5.96– 5.76(m,1H),5.36(d,J=10.2Hz,1H),5.16(d,J=17.0Hz,1H),4.76–4.60(m,2H),4.59–4.4 9(m,1H),4.37(d,J=5.5Hz,2H),4.16(t,J=6.5Hz,1H),3.75(s,3H),3.28–3.05(m,2H).13C NMR (101MHz, CDCl3) δ (ppm) 170.64, 156.16, 143.66, 143.59, 141.41, 135.31, 129.86, 129.78, 127. 95,127.25,125.09,121.37,120.15,119.42,67.27,53.21,52.85,49.13,47.09,26.35.HRMS[M+Na +]:454.1750 (calculated value 454.1737).

[0336] N α -(((9H-fluoren-9-yl)methoxy)carbonyl)-N π -Allyl-L-histidine (N α -Fmoc-(N π -allyl)-L-histidine-OH 105). To a solution of Nα-Fmoc-(Nπ-allyl)-L-histidine-OMe (602 mg, 1.4 mmol) in 27 mL of dioxane, 27 mL of HCl 2M was added and the mixture was refluxed for 10 hours. The solution was neutralized to pH 6 using NaOH 1M. This mixture was extracted twice with DCM. The completion of the extraction process was monitored using TLC. The combined organic phases were washed with brine, dried over MgSO4 and filtered. The organic solvent was removed under vacuum, and the crude product was purified by flash chromatography using DCM-MeOH as eluent (gradient 5% → 20% MeOH + 1% AcOH during 10CV). 450 mg of product was obtained with a yield of 77%. 1 HNMR(400MHz,MeOD-d4)δ(ppm)8.91(s,1H),7.78(d,J=7.5Hz,2H),7.62(t,J=6.8Hz,2H), 7.38(t,J=7.4Hz,2H),7.35–7.25(m,3H),6.04(ddd,J=22.4,10.8,5.6Hz,1H),5.40(d,J=1 0.3Hz,1H),5.24(d,J=17.1Hz,1H),4.87(d,J=4.5Hz,2H),4.53(dd,J=9.4,4.6Hz,1H),4.3 7(d,J=6.7Hz,2H),4.18(t,J=6.6Hz,1H),3.38–3.30(m,1H),3.10(dd,J=15.9,9.6Hz,1H). 13 C NMR(101MHz,MeOD-d4)δ(ppm)173.27,158.40,145.16,145.07,142.60,136.54,132.85,132 .08,128.83,128.16,126.12,120.96,119.36,67.88,53.63,50.24,48.36,26.82.HRMS[M+Na + ]:418.1759 (calculated value 418.1759).

[0337] Synthesis and Characterization of Tyr(OBn) Analogs

[0338] (S)-2-amino-3-(3-cyclopentyl-4-hydroxyphenyl)-propionic acid (cypTyr 107). To a 250 mL round-bottom flask containing L-tyrosine (1 eq., 4.0 g, 22 mmol), 85% H3PO4 (7.8 eq., 20 mL, 173 mmol) and 3 mL cyclopentanol (1.5 eq., 2.9 g, 33 mmol) were added, and the suspension was stirred and heated at 100 ° C overnight. After 16 hours, the reaction mixture was cooled, diluted in 100 mL ice water, and the acid was neutralized with KOH (15.6 eq., 19.3 g, 345 mmol). NaHCO3 was added until pH 5-7 to precipitate the product. The precipitate was filtered and washed with cold water. The solid was dried under a fume hood for 1 day to deliver 4.96 g of crude product (off-white solid) as a mixture of mono-alkylated and di-alkylated tyrosine, which was therefore used in the next step.

[0339] (S) -2-amino -3- (3- cyclopentyl -4- hydroxyphenyl) -propionic acid methyl ester (cypTyr-OMe, 108). Under an inert atmosphere, in a 250mL round-bottom flask, the crude mixture of compound 107 (4.0g) was dissolved in anhydrous methanol (73 equivalents, 45mL, 1.1mol). The solution was cooled to 0 ° C, and then thionyl chloride (3 equivalents, 3.3mL, 46mmol) was slowly added to the flask. The mixture was stirred at room temperature overnight. After 15 hours, methanol was evaporated under reduced pressure, and the mixture was diluted with cold saturated NaHCO (15mL). The product was extracted with three portions of ethyl acetate (EtOAc, each 20mL). The combined organic phase was washed with a portion of brine (20mL), dried over anhydrous MgSO and filtered. EtOAc was evaporated under reduced pressure to obtain 3.85g orange-brown oil. The obtained product was purified using flash chromatography on a normal phase silica gel cartridge with gradient elution 0→7% MeOH / DCM. The mono- and di-alkylated products were completely separated and the mono-alkylated product 15 (1.2 g) was obtained as a clear viscous oil in 26% yield (2 steps). 1 H NMR(400MHz,MeOD-d4)δ(ppm)6.93(d,J=2.0Hz,1H),6.79(dd,J1=8.0,2.0Hz,1H),6.67(d,J=8.4Hz,1H),3.6 7(s,3H),3.64(t,J=6.4Hz,1H),3.32–3.22(m,1H),2.92-2.81(m,2H),2.03–1.94(m,2H),1.86–1.50(m,6H). 13C NMR (101MHz, MeOD-d4) δ (ppm) 176.6, 155.4, 133.8, 128.9, 128.6, 128.4, 116.1, 56.9, 52.5, 41.2, 40.6, 34.2, 34.1, 26.6.

[0340] (S)-2-((tert-butoxycarbonyl)amino)-3-(3-cyclopentyl-4-hydroxyphenyl)-methyl propionate (Boc-cypTyr-OMe 109). Compound 108 (1 equivalent, 1.2 g, 0.7 mmol) was dissolved in a mixture of 30 mL THF / H2O (1:1) in a 250 mL round-bottom flask. NaHCO3 (2 equivalents, 114 mg, 1.4 mmol) was added to the flask and stirred until completely dissolved. Di-tert-butyl dicarbonate (1.5 equivalents, 223 mg, 1.0 mmol) was added and the reaction was run at room temperature for 1 hour. In order to neutralize the solution, 1 M HCl aqueous solution (1.4 mL, 1.4 mmol) was added, and THF was evaporated in vacuo. The final product was extracted from the aqueous phase with three portions of EtOAc (20 mL each). The combined organic phase was washed with a portion of brine (20 mL) and dried over anhydrous magnesium sulfate. The solution was filtered and the solvent was removed in vacuo.The product was purified by flash chromatography using a 20→30% EtOAc / hexanes gradient to afford 16 (1.64 g) as a light yellow oil in 84% yield. 1 H NMR (400MHz, CDCl3) δ (ppm) 6.90 (d, J = 2.0Hz, 1H), 6.77 (dd, J = 8.1, 2.2Hz, 1H), 6.64 (d, J = 8.0Hz, 1H), 4.99 (d, J = 8.2Hz, 1H), 4.54 (dd, J = 13.7, 5.7Hz, 1H),3.71(s,3H),3.28–3.14(m,1H),3.00(d,J=5.2Hz,2H),2.10–1.93(m, 2H),1.86–1.73(m,2H),1.73–1.63(m,2H),1.61–1.51(m,2H),1.42(s,9H). 13 CNMR(101MHz,CDCl3)δ(ppm)172.76,155.33,152.92,132.27,128.07,127.56,1 27.38,115.41,80.14,54.64,52.35,38.94,37.74,33.07,33.02,28.43,25.49.

[0341] (S)-3-(4-(Benzyloxy)-3-cyclopentylphenyl)-2-((tert-butoxycarbonyl)amino)-propionic acid methyl ester (Boc-cypTyr(OBn)-OMe 110). Compound 109 (1 equivalent, 203 mg, 559 μmol) was dissolved in dry acetonitrile (5 mL), and K2CO3 (1.2 equivalents, 92.6 mg, 670 μmol) was added to the mixture, followed by benzyl bromide (1.3 equivalents, 130 mg, 760 μmol). The mixture was refluxed for 18 hours. After the reaction was complete, acetonitrile was evaporated in vacuo. The crude product was redissolved in 15 mL EtOAc and washed with 3 portions of water (15 mL each) and 20 mL of brine. The organic phase was collected, dried over MgSO4, filtered, and the solvent was removed in vacuo. The crude product was purified by flash chromatography using hexane-EtOAc (8:2) as isocratic eluent to afford a clear viscous oil (158 mg) in 58% yield. 1 H NMR (400 MHz, CDCl3) δ (ppm) 7.42–7.24 (m, 5H), 6.95 (d, J = 1.8 Hz, 1H), 6.86 (dd, J = 7.8, 1.5 Hz, 1H), 6.80 (d, J = 8.4 Hz, 1H), 5.04 (s, 2H), 4.95 (d, J = 8.1 Hz, 1H), 4.54 (q, J = 7.7 Hz, 1H), 3.70 (s, 3H), 3.38 (quintet, J = 8.4 Hz, 1H), 3.02 (m, 2H), 2.01 (m, 2H), 1.79–1.50 (m, 6H), 1.42 (s, 9H). 13 C NMR (101MHz, CDCl3) δ (ppm) 172.7,155.8,155.3,137.7,135.3,128.7,128.1,128.0 ,127.9,127.8,127.3,111.9,80.0,70.3,54.7,52.4,39.1,37.8,33.2,28.5,25.7.

[0342] (S)-2-((tert-Butyloxycarbonyl)amino)-3-(3-cyclopentyl-4-(cyclopentyloxy)phenyl)-propionic acid methyl ester (Boc-cypTyr(OCyp)-OMe 111). Using a similar protocol to the synthesis of 110, 111 (640 mg, 1.48 mmol, 54% yield) was obtained from 109 (1.00 g, 2.77 mmol). 1H NMR (400 MHz, CDCl3) δ (ppm) 6.89 (s, 1H), 6.83 (d, J = 7.6 Hz, 1H), 6.71 (d, J = 8.0 Hz, 1H), 5.05–4.84 (m, 1H), 4.82–4.64 (m, 1H), 4.60–4.41 (m, 1H), 3.69 (s, 3H), 3.21 (quintet, J = 7.6 Hz, 1H), 2.99 (d, J = 4.4 Hz, 2H) 2.08–1.28 (m, 25H). 13 C NMR (101MHz, CDCl3) δ (ppm) 172.7, 155.2, 155.0, 135.3, 128.1, 127.1, 126. 9,112.4,79.9,79.1,54.6,52.3,39.5,37.7,33.0,32.9,28.5,25.7,24.2.

[0343] (S)-2-((tert-Butyloxycarbonyl)amino)-3-(3-cyclopentyl-4-propoxyphenyl)-propionic acid methyl ester (Boc-cypTyr(OPr)-OMe 112). Using a similar protocol to the synthesis of 110, 112 (521 mg) was obtained from 109 (900 mg, 2.48 mmol) with a yield of 52%. 1H NMR (600 MHz, CDCl3) δ (ppm) 6.90 (d, J = 1.2 Hz, 1H), 6.84 (dd, J = 8.4 Hz, 1.8 Hz, 1H), 6.71 (d, J = 8.4 Hz, 1H), 4.92 (d, J = 7.8 Hz, 1H), 4.51 (q, J = 7.8 Hz, 1H), 3.87 (t, J = 5.4 Hz, 2H), 3. 69 (s, 3H), 3.28 (quintet, J = 8.4 Hz, 1H), 2.99 (d, J = 6.6 Hz, 2H), 2.01–1.93 (m, 2H), 1.79 (sextet, J = 6.0 Hz, 2H) 1.76–1.70 (m, 2H), 1.68–1.48 (m, 4H), 1.40 (s, 9H), 1.02 (t, J = 7.2 Hz, 3H). 13 C NMR (101MHz, CDCl3) δ (ppm) 172.7, 156.2, 155.3, 135.0, 128.0, 127.4, 127. 3,111.4,80.0,69.8,54.7,52.4,39.3,37.8,33.1,28.5,25.8,23.0,11.0.

[0344] (S) -3- (4- (benzyloxy) -3- cyclopentylphenyl) -2- ( (tert-butoxycarbonyl) amino) -propionic acid (Boc-cypTyr (OBn) -OH 113). To a solution of compound 110 (921 mg, 2.03 mmol) in 10 mL THF was added LiOH (583 mg, 24.4 mmol, pre-dissolved in 10 mL water). The mixture was stirred at room temperature for 3 hours. After completion of the conversion (such as followed by TLC), the pH was adjusted to 5 with HCl 1M, and THF was removed in vacuo. The pH of the aqueous phase was adjusted to 2, and the product was extracted with three portions of ethyl acetate (20 mL each). The combined organic phases were dried with MgSO4, filtered and the solvent was removed in vacuo to obtain a white solid (860 mg) with a yield of 96%. The product was pure enough to proceed to the next step without purification. 1 H NMR (400MHz, CDCl3), δ (ppm) 7.46–7.36 (m, 4H), 7.36–7.29 (m, 1H), 6.95 (d, J = 7.8Hz, 1H), 6.83 (d, J = 8.3Hz, 1H), 5.06 (s, 2H), 4.95 (d, J = 7.9Hz, 1H) ,4.60(dd,J=12.4,5.6Hz,1H),3.50–3.31(m,1H),3.22–2.96(m,2H),2.0 9–1.95(m,2H),1.84–1.72(m,2H),1.71–1.55(m,4H),1.49–1.28(m,10H). 13 C NMR (101MHz, CDCl3) δ (ppm) 177.05, 155.80, 155.46, 137.61, 135.32, 128.63, 128.14, 127.84, 127.77, 127.34,127.26,111.92,80.29,77.48,77.16,76.84,70.23,54.46,39.05,37.23,33.10,28.42,25.61.

[0345] (S)-2-((tert-Butyloxycarbonyl)amino)-3-(3-cyclopentyl-4-(cyclopentyloxy)phenyl)-propionic acid (Boc-cypTyr(OCyp)-OH 114). Using the same protocol as the synthesis of 113, 114 (656 mg, 1.57 mmol, 100%) was obtained from 111 (640 mg, 1.48 mmol). 1H NMR (400MHz, CDCl3) δ (ppm) 6.98 (s, 1H), 6.92 (d, J = 7.9Hz, 1H), 6.75 (d, J = 8.4H z,1H),4.91(d,J=7.9Hz,1H),4.74(p,J=4.0Hz,1H),4.56(d,J=6.6Hz,1H),3.32 –3.17(m,1H),3.16–2.91(m,2H),1.95(dd,J=10.3,5.3Hz,2H),1.90–1.82(m,4 H),1.81–1.71(m,4H),1.70–1.59(m,4H),1.59–1.50(m,2H),1.48–1.28(m,9H). 13 C NMR (101MHz, CDCl3) δ (ppm) 177.03, 155.52, 155.03, 135.42, 128.20, 127.13, 126. 71,112.48,80.29,79.11,54.49,39.56,37.16,33.03,32.85,28.42,25.71,24.16.

[0346] (S)-2-((tert-Butyloxycarbonyl)amino)-3-(3-cyclopentyl-4-propoxyphenyl)-propionic acid (Boc-cypTyr(OPr)-OH 115). Using the same protocol as the synthesis of 113, 115 (467 mg, 1.19 mmol, 93% yield) was obtained from 112 (521 mg, 1.28 mmol). 1 H NMR (400 MHz, CDCl3) δ (ppm) 6.97 (s, 1H), 6.91 (d, J = 8.0 Hz, 1H), 6.72 (d, J = 8 Hz, 1H), 4.94–4.79 (m, 1H), 4.57–4.45 (m, 1H), 3.88 (t, J = 6 Hz, 2H), 3.29 (quintet, J = 8 Hz, 1H), 3.13–2.91 (m, 2H), 2.05–1.89 (m, 2H), 1.85–1.46 (m, 8H), 1.40 (s, 9H), 1.02 (t, J = 7.6 Hz, 3H). 13 C NMR (101MHz, CDCl3), δ (ppm) 177.0, 156.3, 155.6, 135.1, 128.2, 127.4, 127.2, 111.5, 80.4, 69.8, 54.6, 39.4, 37.3, 33.1, 28.5, 25.8, 23.0, 11.0.

[0347] (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-(benzyloxy)-3-cyclopentylphenyl)-propionic acid (Fmoc-cypTyr(OBn)-OH 116). Compound 113 (1 eq, 860 mg, 1.96 mmol) was dissolved in 20 mL of a mixture TFA-DCM (1:1) and stirred at room temperature for 2 hours. TFA and DCM were evaporated in vacuo to provide a green-brown solid. The residual acid was neutralized with a saturated NaHCO3 solution, and 20 mL of THF was added to dissolve the solid residue. NaHCO3 (3 eq, 493 mg, 5.87 mmol) was dissolved in 10 mL of water and added to the mixture, followed by Fmoc-Cl (1.1 eq, 607 mg, 2.07 mmol). The reaction was run at room temperature for 2 hours. THF is evaporated in vacuo, and the aqueous phase is acidified to pH 3 with 1N HCl aqueous solution, extracted with 3 parts of EtOAc (15mL). The combined organic phase is washed with brine and dried with MgSO4. Organic matter is evaporated in vacuo to deliver the crude product. The product is purified by flash chromatography using 0-30% EtOAc+0.25% AcOH / hexane gradient to obtain a light yellow oil (752mg), yield (68%). 1 H NMR (400MHz, CDCl3), δ (ppm) 7.76 (d, J = 7.5Hz, 2H), 7.55 (t, J = 7.9Hz, 2H), 7.46–7.27 (m, 10H), 7. 06(s,1H),6.91(d,J=8.2Hz,1H),6.81(d,J=8.3Hz,1H),5.20(d,J=8.2Hz,1H),5.03(s,2H),4.69( dd,J=13.5,5.8Hz,1H),4.46–4.32(m,2H),4.20(t,J=7.0Hz,1H),3.45–3.31(m,1H),3.17(dd,J=1 4.1,5.3Hz,1H),3.09(dd,J=14.0,6.1Hz,1H),2.01(s,2H),1.82–1.69(m,2H),1.68–1.51(m,5H). 13C NMR (101MHz, CDCl3), δ (ppm) 176.67, 155.94, 143.89, 143.85, 141.42, 137.53, 135.49, 128.65, 127.99, 127.88, 127.42, 127.3 9,127.29,127.24,127.20,125.23,120.13,111.99,70.22,67.33,54.81,47.24,39.13,37.27,33.13,25.62,25.60.HRMS[M+Na + ]584.2405 (calculated value 584.2407).

[0348] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(3-cyclopentyl-4-(cyclopentyloxy)phenyl)-propionic acid (Fmoc-cypTyr(OCyp)-OH 117). Using a similar protocol to the synthesis of 116, 117 (321 mg, 593 μmol, 41% yield) was obtained from 114 (600 mg, 1.44 mmol). 1 H NMR (400MHz, CDCl3) δ (ppm) 7.76 (d, J = 7.5Hz, 2H), 7.56 (t, J = 7.9Hz, 2H), 7.40 (t, J = 7.4Hz, 2H), 7.30(t,J=7.4Hz,2H),7.01(s,1H),6.90(d,J=8.1Hz,1H),6.74(d,J=8.3Hz,1H),5.22(d,J=8.2 Hz,1H),4.79–4.64(m,2H),4.38(t,J=6.9Hz,2H),4.21(t,J=7.0Hz,1H),3.30–2.93(m,3H),2.0 3–1.91(m,2H),1.85(d,J=4.6Hz,4H),1.82–1.71(m,4H),1.70–1.59(m,4H),1.60–1.48(m,2H). 13 C NMR (101MHz, CDCl3) δ (ppm) 176.79,155.97,155.15,143.91,141.41,135.54,128.06,127.84,127.19,126.40,1 25.25,120.10,112.52,79.11,67.33,54.82,47.24,39.63,37.23,33.02,32.86,25.71,25.68,24.17.HRMS[M+H + ]540.2762 (calculated value 540.2745).

[0349] (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(3-cyclopentyl-4-propoxyphenyl)-propionic acid (Fmoc-cypTyr(OPr)-OH 118). Using a similar protocol to the synthesis of 116, 118 (438 mg, 853 μmol, 71% yield) was obtained from 115 (467 mg, 1.19 mmol). 1 H NMR (400MHz, CDCl3) δ (ppm) 7.76 (d, J = 7.6Hz, 2H), 7.55 (t, J = 7.8Hz, 2H), 7.40 (t, J = 7.4Hz, 2H), 7.30 (t, J = 7.4Hz, 2H) ,7.02(s,1H),6.91(d,J=8.2Hz,1H),6.73(d,J=8.3Hz,1H),5.19(d,J=8.2Hz,1H),4.68(dd,J=13.6,5.8Hz,1H),4.43– 4.33(m,2H),4.20(t,J=7.0Hz,1H),3.88(t,J=6.3Hz,2H),3.39–3.24(m,1H),3.16(dd,J=14.1,5.3Hz,1H),3.08(dd,J =14.0,6.2Hz,1H),2.09–1.90(m,2H),1.87–1.71(m,4H),1.70–1.61(m,2H),1.60–1.50(m,2H),1.04(t,J=7.4Hz,3H). 13 C NMR (101MHz, CDCl3) δ (ppm) 176.65, 156.32, 155.97, 143.90, 141.41, 135.11, 127.91, 127.85, 127.34, 127.20, 126. 78,125.24,120.11,111.42,69.65,67.34,54.83,47.24,39.33,37.26,33.02,25.66,25.63,22.89,10.94.HRMS[M+H + ]514.2602 (calculated value 514.2588).

[0350] General protocol for solid phase synthesis

[0351] Following the more specific schemes provided in Examples 2-4, a general scheme for solid phase synthesis of the analogs disclosed herein is provided below.

[0352] The chemistry based on Fmoc is used to synthesize peptides with 0.1mmol scale on solid phase. The first amino acid is loaded into resin using Mitsunobu reaction (Mitsunobureaction). In brief, amino acid (0.3 equivalent, 0.3mmol), triphenylphosphine (3 equivalents, 0.3mmol, 79mg) and 300mg Wang resin are mixed together in 4mL DCM, and continue for 5 minutes. Diisopropyl azodicarboxylate (DIAD, 3 equivalents, 0.3mmol, 59 μL) is dripped, and the mixture is vibrated overnight. Excessive reagent is removed by washing twice with 5mL DCM. Quantitative amino acid loading is carried out by measuring the UV absorbance of dibenzofulvene-piperidine adduct produced by Fmoc deprotection. Load is generally 0.25-0.35mmol / g. When reaching the loading of expectation, resin is covered with 4mL DCM-acetic anhydride-DIPEA (4:1:0.2) solution during 1 hour. The resin is washed with DMF-DCM-iPrOH-DCM-iPrOH-DCM, washed with 5mL of each solvent for 3 minutes (also referred to as the washing scheme). The next amino acid is added to the sequence by 2 steps: 1 / Fmoc deprotection and 2 / amide coupling. The above-mentioned washing scheme is always used to wash the resin between the two steps. Soft protection is achieved by treating the resin with 5mL 20% piperidine / DMF for 10 minutes. For the coupling step, HATU (5 equivalents, 0.5mmol, 190mg) and amino acid (5 equivalents, 0.5mmol) are dissolved in 5mL DMF, transferred to the resin, and then DIPEA (5 equivalents, 0.5mmol, 87μL) is added to start the coupling reaction. The reaction is run for 30 minutes, and excess reagents are removed by filtration. Deprotection and coupling steps are repeated to synthesize linear precursor peptides. The resin is washed using the washing scheme, washed with ether again and dried overnight in a vacuum before the cyclization step.

[0353] Synthesis of Fmoc-N on resin γ -allyl-N γ -nitrobenzenesulfonyl-α,γ-diamino-butyric acid (Fmoc-Alnb-OH). Fmoc-L-Dab(Alloc)-OH was introduced at the His7 position by SPPS and served as the starting residue for the synthesis of the Fmoc-Alnb-containing peptide ( Figure 5). The Alloc group was removed by treating the resin with a solution of Pd(PPh3)4 (0.25 equivalent, 29 mg, 0.025 mmol), phenylsilane (25 equivalent, 311 μL, 2.5 mmol) in DCM under an inert atmosphere for 30 minutes. The resin was washed with 5 mL DCM and then washed with 5 mL DMF for 5 minutes each. In a 20 mL vial, o-nitrobenzenesulfonyl chloride (NsCl, 86 mg, 0.4 mmol) was dissolved in 5 mL NMP and sym-trimethylpyridine (55 μL, 0.4 mmol) was added. This mixture was transferred to the resin and stirred at room temperature for 30 minutes. The resin was filtered and the nosylation reaction was repeated once. The resin was washed with a washing sequence and dried in a vacuum before the allylation step. The allylation step was carried out by Fukuyama-Mitsunobu reaction. In 12mL cylinder, the dried resin was swollen with 4mL anhydrous THF for 5 minutes and filtered. A mixture of allyl alcohol (68 μL, 1mmol), PPh (106mg, 0.4mmol) in 3mL anhydrous THF was poured on the resin and mixed for 5 minutes, then DIAD (0.079mL, 0.4mmol, diluted in 1mL anhydrous THF) was added dropwise. The mixture was stirred at room temperature for 20 minutes. The resin was filtered, and the allylation reaction was repeated once. Conversion was monitored by UPLC-MS. The resin was washed with a washing scheme, and then the peptide extension performed by SPPS was performed.

[0354] Macrocyclization by ring-closing metathesis. Prior to metathesis, Fmoc-L-allylglycine-OH or Boc-L-allylglycine (for truncated analogs) was incorporated at the Pro3 position, while an allyl-containing residue AA (L-allylglycine, N-allylglycine) was introduced at the His7 position during solid phase peptide synthesis. π -allyl-L-histidine or N γ -allyl-N γ -nitrobenzenesulfonyl-α,γ-L-diaminobutyric acid)( Figure 3A ). The general sequence should be Fmoc / Boc-allylglycine

[0355] -Arg (Pbf) -Leu -Ser (OtBu) -AA -Lys (Boc) -Gly-Pro - (C-terminal) -resin. Dry resin (0.1mmol peptide) and Hoveyda-Grubbs catalyst 2nd generation (0.023mmol, 15mg) are added to a dry 10-mL microwave tube. Teflon (teflon) cap is added, and the tube is filled with argon through a needle in a small vacuum chamber by vacuum and backfilling cycles. After argon purge, 4mL of dichloroethane is added, and the reaction mixture is heated at 120 ° C for 10 minutes, 300W in a Discover SP microwave oven (CEM, Matthews, USA). Solvents and reagents are removed by filtration. The resin is washed with a washing scheme. The reaction progress is monitored by LC-MS. If the ratio between the cyclic peptide and the linear peptide is less than 6, the metathesis step is repeated until the cyclic peptide is enriched to the desired amount (cyclic peptide / linear peptide ratio> 6). After washing using the wash protocol, the resin was used for the next step (cleavage of the final product or addition of Pyr-Arg).

[0356] Macrolactamization.Before macrolactamization, Lys (Alloc) or Dap (Alloc) is introduced into Pro3 position, and Fmoc-Asp (OAll) -OH is incorporated into His7 position.On dry resin (0.1mmol peptide), by containing Pd (PPh3) 4 (0.25 equivalent, 29mg, 0.025mmol) and phenylsilane (25 equivalents, 311 μ L, 2.5mmol) DCM treatment resin 30 minutes remove allyl and Alloc protecting groups.Resin is washed with 5mL DCM and 5mL DMF, each 5 minutes.Use DEPBT (5 equivalents, 150mg, 0.5mmol) and DIPEA (5 equivalents, 0.5mmol, 87 μ L) to carry out macrocyclization.Coupling reagent and DIPEA are dissolved in 5mL DMF, then transferred to resin, and cyclization reaction is run overnight. The resin was washed with the wash protocol before proceeding to the next step (Fmoc deprotection / peptide extension).

[0357] Modification after cyclization. Macrocycles 7 and 8 were synthesized from macrocycle 5 on resin. After the metathesis step and the addition of Pyr(Boc)-Arg(Pbf)- at the N-terminus to provide 5 (protecting groups are still there), macrocycle 7 was obtained by deprotecting the o-nitrobenzenesulfonyl group using a mixture of mercaptoethanol (8 equivalents, 57 μL, 0.8 mmol), DBU (5 equivalents, 0.5 mmol, 76 μL) for 15 minutes (repeated once more to ensure complete deprotection). The resin was washed with a washing scheme and then subjected to final cleavage. During the overnight period, macrocycle 8 was synthesized from 7 (with protecting groups) on resin by reductive amination using 37% formaldehyde solution (40 equivalents, 324 μL, 4 mmol), NaBH(OAc)3 (20 equivalents, 423 mg, 2 mmol) in a mixture of THF-TMOF (1:1). Excess NaBH(OAc)3 was quenched with 3 mL MeOH. After gas evolution ceased, the resin was washed with MeOH followed by the wash protocol. It should be noted that a portion of the peptide was cleaved during the reductive amination, which reduced the yield.

[0358] Final cleavage and purification. Final cleavage of the resin and simultaneous removal of protecting groups were performed using a mixture of trifluoroacetic acid (TFA) / triisopropylsilane (TIPS) / water (95:2.5:2.5). The cleavage reaction was run for 2 hours (if the peptide had 0-1 arginine) or 4 hours (if the peptide had 2 arginines). The mixture was filtered through a glass fiber plug to remove solid particles, and the solution was slowly dripped into 30 mL of methyl tert-butyl ether (pre-cooled at 0°C) to precipitate the product. The crude peptide was separated by centrifugation (3000 rpm, 10 minutes), resuspended in 1 mL of 10% acetic acid (AcOH) and allowed to stand for 10 minutes. The two layers were separated: the residual ether layer (top) and the aqueous layer (bottom). The aqueous layer was separated and 1 mL of AcOH 10% was added to extract the residual peptide from the ether layer. This check helps to further clean the mixture and facilitate purification. The aqueous extracts were combined and filtered before purification. The HPLC-MS was carried out on a HPLC-MS system (XSELECT packed with 5 μm particles) from Waters Corporation (Milford, USA) using a binary solvent system (acetonitrile / water + 0.1% formic acid). TM CSH TM The macrocyclic peptide was purified using a Prep C18 column (19 x 100 mm), UV detector 2998, MS SQ detector 2, sample manager 2767 and binary gradient module. Pure fragments (confirmed by UPLC-MS) were pooled and lyophilized to obtain a white solid. A UPLC / MS system from Waters (Milford, USA) was used with an Acquity CSH TMThe purity of the peptides was assessed on a C18 column (2.1 x 50 mm) using the following gradient: acetonitrile and water containing 0.1% HCOOH (0→0.2 min: 5% acetonitrile; 0.2→1.5 min: 5%→95%; 1.5→1.8 min: 95%; 1.8→2.0 min: 95%→5%; 2.0→2.5 min: 5%). The purity of all peptides was >95% except for 21 (91%), 24 (93%), 32 (93%), and 33 (90%). HRMS spectra were obtained using electrospray infusion using a maXis ESI-Q-Tof instrument from Bruker (Billerica, USA).

[0359] Cell culture

[0360] HEK293 cells stably expressing YFP-tagged human APJ were cultured in DMEM medium supplemented with 10% FBS at 37°C in a humidified atmosphere maintained at 5% CO2. The antibiotic G418 (400 μg / mL) was added to maintain the selection pressure for cells expressing APJ, while penicillin / streptomycin (0.1%) was used to prevent bacterial contamination.

[0361] Combination experiment

[0362] Binding experiments were performed on cell membranes of HEK293 cells stably expressing YFP-tagged human APJ receptors. Cells were stored frozen at -80°C and thawed quickly (1 min at 37°C) immediately before the experiment. The thawed cells were resuspended in 5 mL of EDTA solution (1 mM EDTA, 50 mM Tris-HCl, pH 7.4), transferred to a 10-mL falcon tube, and centrifuged at 3500 g for 15 minutes at 4°C to extract the cell membrane. The precipitate (cell membrane) was suspended in binding buffer (50 mM Tris-HCl, 0.2% BSA, pH 7.4). The assay was run in a 96-well plate. Fifteen μg of membrane protein was mixed with 0.2 nM radiolabeled [ 125 I][Nle75,Tyr77][Pyr 1 ]-Apelin-13 (820Ci / mmol) 3 Incubate together and test a concentration range of 10 in a total volume of 200 μL at room temperature. -5 Up to 10 -11The incubation mixture was filtered through a glass fiber filter (Millipore, pre-absorbed with 0.5% PEI at 4°C for 2 hours) to remove unbound ligand, and the filter membrane was washed three times with 170 μL of cold binding buffer (4°C). Gamma emission was measured using a 1470 Wizard gamma counter from PerkinElmer (Waltham, USA) (80% efficiency). Nonspecific binding did not exceed 5% of the total signal (by comparing with 10 -5 IC was determined from these results using GraphPad Prism 8 50 The value represents the concentration of test ligand that displaces 50% of the radiolabeled ligand from the receptor. [Pyr 1 ]-K of Apelin-13 D is 1.8 nM. Using the Cheng-Prusoff equation according to IC 50 Calculate the dissociation constant K i The results are shown as the mean ± SEM of two to three independent experiments, each performed in duplicate (Yung-Chi et al., 1973).

[0363] For Gα i1 BRET assay for activation and β-arrestin2 recruitment

[0364] HEK293 cells were cultured in high glucose DMEM medium with 10% FBS, 100 U / mL penicillin / streptomycin, 2 mM glutamine and 20 mM HEPES at 37°C in a T175 flask with 5% CO2 in a humidified chamber. After 24 hours, PEI (Murza et al., 2015; Galès et al., 2006; Zimmerman et al., 2012) was used to infect the cells with proteins encoding human APJ, Gα i1 -RlucII(91), GFP10-Gγ2 and G β1 (For BRET-based Gα i1Cells were transfected with plasmids encoding APJ-GFP10 and RlucII-β-arrestin2 (for BRET-based β-arrestin2 recruitment assay). Prior to the assay, cells were transferred to white 96-well plates from BD Bioscience (Mississauga, Canada) at a concentration of 50,000 cells / well for 24 hours and incubated overnight at 37°C. The cells were then washed with phosphate-buffered saline (PBS) and 90 μL of Hanks' balanced salt solution was added to each well. The plates were then incubated at 37°C (Gα i1 ) with a concentration of 10 -5 M to 10 -11 Cells were stimulated with analogs in the M range for 5 min or at room temperature for 30 min (β-arrestin 2). After stimulation, 5 μM coelenterazine 400A was added to each well and the BRET assay was performed using a GeniosPro plate reader (Tecan, Austria). 2 Filter set read plate. BRET ratios were determined for GFP10 em / RlucII em Data were plotted and EC was determined using GraphPad Prism 8 50 Each data point represents the mean ± SEM of at least three different experiments, each performed in triplicate.

[0365] Rat plasma stability

[0366] Plasma was obtained from male Sprague-Dawley rats by collecting blood in heparin tubes and centrifuging at 13,000 rpm to remove blood cells. The separated plasma was stored at -80 ° C and thawed before testing. In a 96-well plate, 6 μL of a 1 mM peptide solution was incubated with 27 μL of plasma at 37 ° C in an oven equipped with an orbital shaker. The wells were sealed with a tight-fitting lid to avoid water evaporation during incubation. At 0, 1, 2, 4, 6 and 24 hours, the plasma was inactivated with 140 μL ACN-EtOH (1: 1) solution containing 0.25 mM N, N-dimethylbenzamide (internal standard), and the wells were sealed again with a tight-fitting lid. At 24 hours, when all plasma was inactivated, the lid was removed and the mixture was transferred to a 96-well filter plate Impact TMProtein precipitation (Phenomenex, California, US). A 96-well UPLC plate was placed at the bottom to collect the sample. Both plates were centrifuged at 500 g for 10 minutes at 4°C to accelerate filtration. The collected filtrate was diluted with 80 μL of water and analyzed on an Acquity UPLC-MS system Class H (Acquity Protein BEH C4 column (2.1 x 50 mm), 1.7 μm particles, pore size ). The amount of remaining peptide was plotted as an exponential monophasic decay curve using GraphPad Prism 8, which allowed the calculation of peptide half-life. The results are presented as the mean ± SEM of at least 3 independent experiments, each performed in a simple manner.

[0367] In vivo pharmacokinetics

[0368] Male Sprague-Dawley rats aged 8-10 weeks were used in this study. Twenty-four hours before the experiment, a jugular vein catheter ( Laboratory tubes; 0.02 ID x 0.037 OD) were used for intravenous injections (iv, 3 mg / kg, for 0.9% saline solution containing mimetic 42, 43, or Apel3, ≈350 μL) and for blood collection. Prior to iv injection, animals were placed in a closed chamber to facilitate blood sampling. After iv administration, blood samples were collected at 5, 10, 30, 60, 120, and 240 minutes (for [Pyr 1 Blood samples (0.2 mL, corresponding to 0.1 mL of plasma after centrifugation) were collected in K2-EDTA microcontainer tubes (Sarstedt, Nümbrecht, Germany) at 1, 2, 5, 10, 15 minutes for 1 h (1, 2, 5, 10, 15 minutes for 1 h) and 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 67, 68, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 111, 112, 113, 114, 115, 126, 131, 1

[0369] Sample preparation

[0370] Peptides are extracted using a combination of protein precipitation and solid phase extraction steps. Plasma samples are thawed on ice. After vortexing (60 seconds), 100 μL samples are extracted and 300 μL cold acetonitrile is added to precipitate plasma proteins. The sample is then vortexed (60 seconds) and centrifuged at 4500 rpm for 10 minutes at 4°C. The supernatant is then separated and passed directly through an HLB extraction column (prime) for additional cleaning. The filtrate is diluted 10 times in 0.1% formic acid / water and filtered through a 0.22 μm syringe filter and then analyzed by LC / MS / MS.

[0371] Mass spectrometry analysis

[0372] Samples were analyzed on a Sciex Qtrap 6500+ equipped with microflow liquid chromatography (Eksigient M3 microflow) and a UPLC HSS-T3 column (1 mm x 100 mm, 1.8 μm, equipped with a 0.2 μm sintered prefilter). The solvent flow rate was set to 50 μL / min, the column temperature was maintained at 40°C, and the injection volume was 3 μL. The mobile phases were 0.1% formic acid / water (A) and 0.1% formic acid / acetonitrile (B). The elution gradient started with 2% eluent B, increased to 95% in 8 minutes, maintained at 95% for 2 minutes, and then returned to the initial conditions in 2 minutes, with a total run time of 13 minutes. Optimized parameters for peptide fragmentation were obtained by direct infusion of Ape13, 42 and 43 analytical standard solutions at 100 ng / mL. The analysis used two subtraces (transitions), the most abundant of which was used for quantification and the second most abundant for confirmation.

[0373] In vivo blood pressure measurement

[0374] animal

[0375] Adult male Sprague-Dawley rats aged 8-10 weeks (Charles River Laboratories, St-Constant, Quebec, Canada) were housed under a 12-hour light / 12-hour dark cycle with free access to food and water. Animal experimental protocols were approved by the Animal Care Committee of Université de Sherbrooke and were in accordance with the policies and directives of the Canadian Council on Animal Care.

[0376] Blood pressure test

[0377] Male Sprague-Dawley rats (8-10 weeks old) were anesthetized with ketamine / oxazine injection (87 / 13 mg / kg intramuscular injection) and placed in a supine position on a thermostatic pad. The right carotid artery was cannulated with PE 50 (filled with heparinized saline) and connected to a Micro-Med transducer (model TDX-300, Calabasas, USA) and a Micro-Med blood pressure analyzer (model BPA-100c). Vehicle (isotonic saline) was administered by iv bolus injection, followed by injection of Apel3, compound 9, 20, 29, 42 or 43 (given at 19.5 and 65 nmol / kg; volume of 0.25 mL within 10 seconds) through another catheter (PE10) inserted into the left jugular vein 5 minutes later. This iv catheter was flushed with saline (0.2 mL) immediately after each injection.

[0378] Echocardiography

[0379] Transthoracic echocardiography was performed using a Vevo 3100 ultrasound device in Sprague-Dawley rats anesthetized with isoflurane (2%; 1.5 ml / min; Baxter) using an MX250 transducer (FUJIFILM, VisualSonic, ON, Canada) before (baseline) and 3, 6, and 24 hours after subcutaneous injection of peptide (0.2 and 2 μmol / kg). Two-dimensional short-axis views of the LV were obtained at the level of the papillary muscles, and M-mode tracings were recorded. Heart rate (HR) was calculated from these images, and LV end diastolic pressure (LVEDd) and LV end systolic diameter (LVESd) were measured by the leading edge method according to the American Society of Echocardiography guidelines. Fractional shortening (FS) was calculated by the following formula: FS = ([LVEDd-LVESd / LVEDd] x 100%). Cardiac output (CO) was assessed from the LV long-axis view. According to the Simpson method, stroke volume (SV) is calculated by tracing the endocardial border at end-systole and end-diastole, and CO is obtained as CO = SV x HR.

[0380] Example 2 : Synthesis of the apelin peptide 13 analogs of Table 1

[0381] Analogs of 97 were designed and synthesized using various types of macrocyclic linkers, such as saturated hydrocarbon chains (13), lactam groups (14, 17), histidine mimetics (15), sulfonamides (16), secondary amines (18), and tertiary amines (19). Figure 2A ).

[0382] The precursor linear peptide was synthesized using classical solid phase peptide synthesis (SPPS) and Fmoc chemistry. To construct the macrocycle, the Pro3 and His7 residues have been replaced with unnatural amino acids as part of the linker. For compound 97, allylglycine residues were introduced at two positions to prepare for cyclization using ring closing metathesis (RCM) ( Figure 3A Compound 13 was obtained from 97 by hydrogenation using a 10% Pd / C catalyst (Green et al., 2013). Compounds 14, 17, 46, and 47 were synthesized by macrolactamization according to a previously described protocol (Alcaro et al., 2004), where Dap(Alloc) or Lys(Alloc) was located at the Pro3 position, respectively, and Asp(OAll) was located at the His7 position. Macrocycles 15, 16, 18, and 19 were also prepared using RCM ( Figure 3B For this purpose, allylglycine was introduced at the Pro3 position and N π -allyl-histidine (15'Alh) or N γ -allyl-N γ -nitrobenzenesulfonyl-α,γ-diamino-butyric acid (Alnb of 16, 17, 18) was placed at the His8 position. et al., 2018), the Pro3→allylglycine mutation is located between Arg2 and Arg4, which makes it impossible for the peptide to cyclize after the introduction of Arg2, probably due to steric hindrance and catalyst chelation by the arginine residue. Therefore, cyclization was performed before adding the Pyr1-Arg2 moiety ( Figure 3B ).

[0383] The conversion of the RCM step was generally >50%, however, in some cases, such as compound 14, the yield was at most about 20-25%, and due to the presence of N π -allyl-histidine, which required longer heating time (100°C, 2 hours). A possible explanation is that the imidazole ring of histidine can act as a chelating agent, thereby poisoning the Hoveyda-Grubbs catalyst.

[0384] Intermediate Fmoc-LN π -allyl-histidine-OH (Fmoc-Alh-OH) was prepared from Fmoc-L-His(Trt)-OH 102 in three steps ( Figure 4 The key step is to use in situ generated allyl triflate to make the N- π The sterically hindered trityl (Trt) protecting group was maintained at the N τ position, thus allowing N πIn the next step, the methyl ester 104 was hydrolyzed using 2M HCl in dioxane (1:1) under reflux to afford Fmoc-LN π -allyl-histidine-OH 105.

[0385] The synthesis of analogs 16, 18, and 19 required the residue Fmoc-Alnb-OH ( Figure 3B ), which is easily prepared on the solid phase from Fmoc-Dab(Alloc)-OH. The Alloc protecting group was selectively removed using Pd(PPh3)4 and PheSiH3 as scavengers. Nitrobenzenesulfonylation of the γ-amine group with o-nitrobenzenesulfonyl chloride and sym-collidine followed by allylation using the Fukuyama-Mitsunobu reaction afforded a peptide containing Fmoc-Alnb ( Figure 5 ).

[0386] To reduce the size of the Ape13 analogs, the N- and C-termini of the macrocyclic analogs were gradually truncated. The truncated peptides were synthesized using the same protocol as above ( Figure 3A -B). In the final series, the C-terminal Nle11 residue was replaced by a non-natural amino acid. Among the non-natural residues, several Tyr(OBn) analogs have been included, such as cypTyr(OBn), dcypTyr(OBn), cypTyr(OPr), and cypTyr(OCyp), because the incorporation of Tyr(OBn) was previously found to increase the affinity for the binding pocket (Murza et al., 2015). In previous studies, cyclopentyl (Cyp) was found to affect the binding and signaling profiles of peptides containing Tyr(OBn) ( et al., 2021). Under reflux conditions, 2 equivalents of 85% phosphoric acid containing cyclopentanol were used to introduce it onto tyrosine (106) to form precursor 107 ( Figure 6 This intermediate was then subjected to a series of transformations (esterification of 107, Boc protection of 108, alkylation of 109, ester hydrolysis of 110-112, Boc removal and Fmoc protection of 113-115) to provide Fmoc-cypTyr(OBn)-OH (116), Fmoc-cypTyr(OCyp)-OH (117), and Fmoc-cypTyr(OPr)-OH (118) suitable for SPPS.

[0387] Compounds 20-45, 48-59 were prepared using a method similar to that used to synthesize 97. In brief, the first amino acid was loaded onto Wang resin using the Mitsunobu reaction (loading 0.3 mmol / g). Linear peptides were synthesized using Fmoc-based chemistry and macrocyclized with Hoveyda-Grubbs catalyst II (120° C., 10 minutes). Non-natural amino acids carrying terminal olefins, such as Lys(N-butenyl), Lys(N-All), Orn(N-butenyl), Dab(N-butenyl), were prepared on resins from Lys(Aloc), Orn(Aloc), and Dab(Aloc) using chemistry similar to that described above for the synthesis of Fmoc-Alnb-OH.

[0388]

[0389] For modification at the N-terminus, as in compounds 21, 23, 4, 47, 49, the Fmoc protecting group was removed after cyclization and the free amino group was derivatized by acetylation or guanidinization (using 1H-pyrazole-1-carboxamidine hydrochloride, CAS: 4023-02-3). Compounds 15, 16, 20, 28-29 and 34-45 are described in Figure 7-8 .

[0390] Example 3 : Synthesis of apelin peptide 17 analogs in Table II

[0391] Step 1: Loading into resin 2-chlorotrityl 400 mg (loading 0.35 mmol / g).

[0392] First, the resin is swollen and washed with DCM. Amino acid and DIPEA2.5 equivalents are dissolved in 4mL DCM, and this solution is poured on the resin. The mixture is mixed overnight. 5mL mixture DCM-MeOH-DIPEA (7:2:1) is used to end-cap unreacted 2-chlorotrityl chloride. After each reaction (end-capping, Fmoc deprotection, amide coupling), the resin is washed with DMF-DCM-iPrOH-DCM-iPrOH-DCM, each solvent for 3 minutes.

[0393] Step 2: Amino Acid Coupling

[0394] Fmoc was removed by treating the resin with 20% piperidine in DMF for 10 minutes. The resin was drained and the deprotection step was repeated once. The next amino acid was added by reacting the free N-terminal amine with 5 equivalents of the corresponding Fmoc-protected amino acid (5 equivalents), HATU and 5 equivalents of DIPEA. Glu (OAll) and Lys (Alloc) were incorporated into their corresponding positions on the peptide sequence.

[0395] Step 3: Deprotection of Alloc and Allyl

[0396] The dried resin was transferred to a 10-mL microwave tube and swollen in 5mL DCM. The mixture was sealed with a lid and bubbled under argon for 10 minutes, then PheSiH was added. When the lid was slightly opened and the argon flow was increased, tetrakis(triphenylphosphine)palladium (Pd(Ph)) was added to the reaction mixture. The mixture was bubbled with argon for 2 minutes and stirred at room temperature for 30 minutes. The resin was washed with a washing sequence: DMF-DCM-MeOH-DCM-MeOH-DCM (3 minutes, 5mL of each solvent).

[0397] Step 4: Peptide cyclization

[0398] To a solution of 5 equivalents of DEPBT in 4 mL of DMF was added DIPEA. This solution was transferred to the reactor containing the resin and the mixture was shaken overnight.

[0399] Ac-Lys(Boc)-Phe-Arg(Pbf)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Pro-Arg(Pbf)-Leu-c[Glu-HIs(Trt)-Lys(Boc)-Lys]c-Pro-Nle-Pro-cypTyr(OBn)-resin (precursor to analog 76).

[0400] Step 5: Final cleavage, deprotection and purification

[0401] Prepare 5mL cleavage mixture of TFA-TIPS-H2O (95:2.5:2.5) and mix thoroughly. Transfer resin to 20-mL vial and add cleavage mixture. Stir this mixture for 5 hours. Filter resin out, and filtrate is added dropwise in precooled TBME to precipitate peptide. Suspension is centrifuged to pull down solid (3000rpm x 10 minutes at 4 ℃). Remove supernatant, and evaporate residual ether under weak air flow, continue 30 minutes. Gained solid is dissolved in 1900 μL 10% acetic acid solution, and is filtered into LC-MS preparative vial (maximum 3mL) by PTFE 0.22um filter. Use gradient 10-25% ACN (+0.1% formic acid) to purify peptide in 15 minutes on preparative HPLC-MS. Pure fraction is freeze-dried to provide 3mg white powder (analog 76).

[0402] Example 4 : Synthesis of Elabela analogs of Table III

[0403] Material

[0404] Fmoc-protected (L)-amino acids, 2-chlorotrityl chloride resin, and [O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate] (HATU) were purchased from Matrix Innovation (Canada). N,N-diisopropylethylamine (DIPEA) and unnatural amino acids were purchased from Chem Impex (USA). Piperidine was purchased from ACP (Canada). All other solvents were purchased from Sigma-Aldrich (Canada) or Fisher Scientific (USA) and were of the highest commercially available purity. All reagents and starting materials were used as received. Symphony TM Peptide elongation was performed using X-Peptide Synthesizer.

[0405] Step 1: Loading of 2-chlorotrityl chloride resin

[0406] To load the first amino acid of the sequence, 2-chlorotrityl chloride resin (0.25mmol / g, 400mg) was treated with dichloromethane (DCM, 4mL) containing an Fmoc-protected amino acid (1 equivalent), N,N-diisopropylethylamine (DIPEA, 2 equivalents). The mixture was shaken on an orbital shaker at room temperature for 2 hours, and then the resin was washed with DCM (2x 5mL), 2-propanol (1x 5mL), DCM (1x5mL), 2-propanol (1x 5mL), DCM (2x 5mL) in sequence for 3 minutes. Then a capping solution of DCM / MeOH / DIPEA (7 / 2 / 1, 5mL) was added, and the mixture was shaken at room temperature for 1 hour and washed with the above solvent sequence.

[0407] Step 2: Peptide extension

[0408] Peptide synthesis is carried out with typical Fmoc solid phase peptide synthesis (SPPS) program.2-Chlorotrityl chloride resin (0.25mmol / g, 400mg, loaded with the first amino acid of sequence) is placed in peptide synthesizer reactor and swollen with N,N-dimethylformamide (DMF) (3x 6 minutes, 4.5mL).It should be noted that the resins during coupling, deprotection and washing steps are mixed by N2 bubbling.Then the Fmoc group is deprotected with 20% piperidine / DMF (2x 5 minutes, 4.5mL), then the amino acid (5 equivalents) of subsequent Fmoc protection is connected in DMF / NMP (4.5mL) in the presence of HATU (5 equivalents), DIPEA (10 equivalents), and the reaction is carried out for 30 minutes.Then piperidine (20% in DMF) is used in each step to deprotect the Fmoc group. After each coupling and Fmoc deprotection step, the resin was washed with DMF (4 x 1 min 30 sec, 4.5 mL).

[0409] Step 3: Allyl / Aloc deprotection

[0410] In a typical procedure, after coupling the last amino acid of the sequence, the allyl / Aloc protecting group was selectively deprotected with tetrakis(triphenylphosphine)palladium (Pd(PPh3)4) (0.2 eq) and phenylsilane (PhSiH3) (20 eq) in argon degassed DCM (5 mL) and the reaction was carried out during 30 minutes. The resin was then washed with DMF (3x 1 min 30 sec, 4.5 mL) and DCM (5x 6 min, 4.5 mL).

[0411] Step 4: Macrolactamization-cleavage / deprotection

[0412] Then, macrolactamization was performed with 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazine-4(3H)-one (DEPBT) (5 eq.) and DIPEA (5 eq.) in DMF (5 mL) over 16 hours. After resin washing (DMF, 5 x 1 min 30 sec., 4.5 mL), the macrocycle was cleaved from the resin and the protecting groups were removed with a mixture of TFA (trifluoroacetic acid) / H2O / TIPS (triisopropylsilane) 95 / 2.5 / 2.5, v / v (2 mL / 0.2 g resin) at room temperature for 4 hours. The crude product was precipitated in tert-butyl methyl ether (TBME) at 0°C, centrifuged and the supernatant removed, or the crude product was directly evaporated under vacuum. The crude product was then redissolved in 7:3H2O / acetonitrile (ACN) and lyophilized before purification by reverse phase HPLC.

[0413] Step 4: Ring Closing Metathesis (RCM) - Cleavage / Deprotection

[0414] Then, ring closing metathesis (RCM) was performed in a CEM microwave at 50°C with Hoveyda Grubbs 2nd generation catalyst (0.2 eq) and benzoquinone (1 eq) in DCE (4 mL) for 1 hour. The resin was then washed with DCM (3x), MeOH (3x) and DCM (3x) and dried before the cleavage step. The resin and protecting groups were removed at room temperature with a mixture of TFA (trifluoroacetic acid) / H2O / TIPS (triisopropylsilane) 95 / 2.5 / 2.5, v / v (2 mL / 0.2 g resin) for 4 hours. The crude product was precipitated in tert-butyl methyl ether (TBME) at 0°C, centrifuged and the supernatant removed, or the crude product was directly evaporated under vacuum. The crude product was then redissolved in 7:3H2O / acetonitrile (ACN) and lyophilized, and then purified by reverse phase HPLC.

[0415] Step 5: Purification and characterization

[0416] The crude product was resuspended in 7:3 H2O / acetonitrile (ACN) and analyzed on a preparative HPLC-MS system (XSELECT packed with 5 μm particles) from Waters Corporation (Milford, USA). TM CSH TM Purification was performed on a UPLC-MS system (Acquity 1000 filled with 1.7 μm particles) from Waters Corporation (Milford, USA). CSH TM Compounds were analyzed on a C18 column (2.1 x 50 mm) using the following gradient: acetonitrile and water containing 0.1% HCOOH (0→0.2 min: 5% acetonitrile; 0.2→1.5 min: 5%→95%; 1.5→1.8 min: 95%; 1.8→2.0 min: 95%→5%; 2.0→2.5 min: 5%).

[0417] Synthetic schemes for the compounds of Table III are also presented in Fig. 9A -B, 10 and 11A-B.

[0418] Example 5 : Characterization of the apelin peptide 13 analogs in Table I

[0419]

[0420]

[0421]

[0422] Example 6: Characterization of the apelin 17 analogs of Table II

[0423]

[0424]

[0425] Example 7 : Characterization of Elabela analogs of Table III

[0426]

[0427]

[0428]

[0429] Example 8 :Binding affinity and other biological properties of apelin peptidergic compounds

[0430] Various properties of the apelin peptidergic compounds of the present disclosure were determined and are presented in Tables I-III below.

[0431] Dissociation constant K i Reflects the binding affinity of the ligand to its receptor (K i Binding (nM)) and corresponds to the concentration of ligand that displaces 50% of the radiolabeled pyr-apelin-13. 125 I][Nle 75 ,Tyr 77 ][Pyr 1 ]-Apel3, which was measured by competitive binding assay on membranes prepared from HEK293 cells stably expressing human APJ (hAPJ).

[0432] EC 50 Gαi1 measurements determined the induction of Gα by BRET-based biosensors in HEK293 cells expressing the hAPJ receptor i1 The concentration of ligand that activates 50% of the maximal response.

[0433] EC 50 β-arr2 measurements determined the concentration of ligand that induced 50% maximal response of β-arr2 recruitment by a BRET-based biosensor in HEK293 cells expressing the hAPJ receptor.

[0434] The in vitro half-life data represent the proteolytic stability of the analogs after incubation in rat plasma at 37°C for several time points up to 24 hours. The percentage of analog remaining was calculated by taking the ratio between the AU of the compound and the AUC of the internal standard. The half-life was extrapolated from the curves.

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441] Example 9 : Apelin 13 analogs that bind to mutant APJ receptors

[0442] The extracellular surface of the APJ receptor has several negatively charged residues on its N-terminal tail and extracellular loops, such as E20, D23, D92, D94, D172, E174, D184, and E194 (Ma et al., 2017). To investigate their role in receptor binding of macrocyclic analogs, the affinity of compounds 20 (N-terminal acetylation), 22 (absence of the N-terminal amine), and 24 (Arg4Nle) was determined on the receptor with APJ E20A and D23A mutations, as these mutations were previously shown to be potential binding sites for the cationic portion of apelin peptides (Table IV). The results showed that the affinity of 20, 22, and 24 was less affected by these mutations (changes <2.5-fold) compared to the affinity of Apel13 (2.4 to 7.7-fold decrease), indicating that E20 and D23 of APJ may not play an important role in the binding of these macrocyclic analogs, but they may be important in conferring higher affinity to compounds with positive charges in the N-terminal portion.

[0443] Table IV. Binding affinity of compounds 20, 22, and 24 to APJ receptor mutants

[0444]

[0445] Example 10 :Functional activity of apelin 13 analogs

[0446] Test K iAll apelin 13 analogs were <20 nM in their ability to activate downstream signaling pathways of the APJ receptor. To this end, a BRET-based biosensor was used to monitor the expression of G proteins (Gα i1 ) activation and β-arrestin 2 recruitment. Notably, this analysis revealed that some macrocycles acted as partial agonists, whereas others showed biased signaling for some of the pathways studied ( Fig.12 , panels AB, Table V).

[0447] Table V. Affinity, functional activity and plasma stability of Ape13 macrocyclic analogs

[0448]

[0449] Example 11 :In vitro plasma stability

[0450] The plasma stability of compounds with an affinity for the APJ receptor of less than 20 nM was evaluated (Table V above). First generation macrocyclic analogs 97, 13, 15, 16, 18 and 19 showed good stability in rat plasma, showing t 1 / 2 The range is 2 to 5 hours, which is much higher than Ape13 (t 1 / 2 0.4 hours). Peptides with polar groups on the linker (18, 19t 1 / 2 2.0-2.2 hours) showed a lower half-life (for 97, 13, 15, 16t 1 / 2 4.0-4.7 hours), which suggests that the linker affects the stability of the macrocycle.

[0451] Compared to compound 97, truncation of the N-terminal tail (Pyr1-Arg2) slightly reduced plasma stability, probably due to exposure of the N-terminal amine, making the compound more susceptible to aminopeptidases. However, analogs 20, 23, and 28 still showed half-lives of less than 3 hours. Macrocyclic analogs are always more stable than their linear analogs. Somewhat surprising was the effect of C-terminal truncation (Pro12-Phe13 removal), which also reduced peptide stability. In fact, analog 29 (t 1 / 2 The stability of 20 (t 0.9 h) was one-third that of 20, which was truncated only at the N-terminus, and was 1 / 3 that of the full-length analog 13 (t 1 / 2 4.7 hours). The C-terminus of Apel3 is known to be cleaved by metalloproteinases such as ACE2 and PRCP at the penultimate position (Yang et al., 2017). However, this cleavage site is removed in these truncated analogs.

[0452] The peptide stability of analog 29 was improved by introducing a D amino acid at the C-terminal Nle11 position. D amino acids are not normally used by the body, and proteases have not evolved for their recognition (Feng et al., 2016), which explains why macrocycles 42, 43, 44, 45 carrying D-1Nal, D-2Nal, D-Tyr(OBn), and D-Tyr substitutions, respectively, are much more stable than the parent compound 29, with half-lives ranging from 2.4 to >24 hours. 43 is the most stable compound in this series, showing a half-life of >24 hours.

[0453] Example 12 : In vivo pharmacokinetics

[0454] The most potent truncated macrocycle (42) and the most stable analog (43) were selected for in vivo pharmacokinetic analysis. The compounds were administered intravenously to rats at 3 mg / kg via the jugular vein, and blood was drawn at 5, 10, 15, 20, 30, 60, 120, and 240 minutes after injection for subsequent LC / MS-MS analysis ( Fig.13 ). As expected, 42 and 43 were stable and detected in rat plasma up to 2 hours after injection, while Ape13 completely disappeared after 5 minutes. Compound 42 showed a half-life of 24 minutes and a plasma clearance of 2.29 ml / min / kg (Table VII). In particular, analog 43 had an in vivo t1 / 2 of 220 minutes, resulting in a circulating concentration of 8.6 μg / mL at 4 hours after injection (compared to 36.1 μg / mL at 5 minutes). This compound has a long half-life and a low plasma clearance (0.34 ml / min / kg), revealing the possibility of using Ape13 analogs as cardioprotective drugs in a single bolus injection.

[0455] Table VII. Pharmacokinetic profiles of macrocycles 42 and 43.

[0456]

[0457] Example 13 : Effects on blood pressure

[0458] The inventors evaluated the ability of compounds 15, 20, 29, 42 and 43 to regulate blood pressure in rat plasma. The effects on blood pressure were evaluated at two doses, 19.6 and 65 nmol / kg. The dose of 19.6 nmol / kg corresponded to the maximum effect of Apel3, while the higher dose of 65 nmol / kg was selected to determine whether less potent analogs could have an effect.

[0459] As the size of the truncated analogue decreased, it gradually lost its effect on blood pressure and lost its ability to recruit β-arrestin 2 (Besserer-Offroy et al., 2018). 50 33 nM, Emax 115%) produced a similar decrease in blood pressure as Apel3 (ΔMABP -40 mmHg), and the response lasted slightly longer, which may be due to a longer half-life and higher efficacy in the recruitment of β-arrestin 2 ( Fig.14 In contrast, analog 20 (β-arr2EC) with a truncated N-terminal tail did not inhibit the expression of β-arr2 in the presence of β-arr2. 50 143nM, E max 98%) did not reach the same response value as Ape13 ( Fig.14 Similarly, analog 29 (β-arr2EC) with both N-terminal and C-terminal truncations 50 743nM, Emax 69%) had little effect on blood pressure (ΔMABP-13mmHg), while 43 (β-arr2EC 50 Compound 42 induced a smaller drop in blood pressure than Apel3 (ΔMABP -24 mmHg), despite having no effect on APJ binding (31, K i 0.6nM for Ape13, K i 0.6 nM) and recruitment of β-arrestin 2 (42, EC 50 31nM for Ape13, EC 50 37 nM) had similar potency. The difference could be explained by the lower maximal potency of 42 for β-arrestin 2 recruitment (E max 70%), indicating its partial agonist activity on this pathway. Similarly, compound 43 was only partially effective on the b-arrestin 2 pathway (E max 55%) and lower potency (43, EC 50 232 nM), which most likely explains its lack of efficacy on blood pressure.

[0460] Example 14 : Effects on Cardiac Performance

[0461] The inventors used echocardiography to study the cardiac effects of macrocycles 42 and 43, which showed small size, good affinity for APJ, and improved in vitro and in vivo half-life. It should also be noted that both compounds are Gα i1Full agonists with good potency, while they are all partial agonists against β-arrestin, but 42 is more potent than 43 on these pathways (6.2 to 7.5 times).

[0462] To demonstrate whether peptides with higher stability can maintain cardiovascular effects after a single bolus administration, Apel3 and two macrocycles 42 and 43 were administered to rats by subcutaneous injection (sc) at two doses: 0.2 μmol / kg (low) and 2 μmol / kg (high, for macrocycles, close to 2 mg / kg). Left ventricular fractional shortening (FS), an indicator of cardiac contraction, and cardiac output (CO), an indicator of overall cardiac performance, were monitored ( Fig.15A -B). Three hours after injection, no observable effect of Apel3 was detected at 0.2 μmol / kg, whereas 42 showed a significant increase in FS only, and 43 showed a significant increase in both FS and CO, consistent with its longer half-life.

[0463] At the highest tested dose of 2 μmol / kg, Ape13, 42, and 43 all showed significant improvements in FS and CO of up to 22-27% relative to baseline at 3 hours after injection. Therefore, previous studies have confirmed that very high doses of Ape13 (50 mg / kg or approximately 32 μmol / kg, sc) can help overcome its short half-life and ensure therapeutically effective concentrations (Onorato et al., 2019). However, most of the compounds tested lost their potency by 6 hours after injection due to metabolism and elimination. At the highest dose (2 μmol / kg), only 43 still showed significant effects on FS (17% increase relative to baseline) and CO (16% increase relative to baseline) at 6 hours. These results are consistent with 42 (in vivo t 1 / 2 24 minutes) and 43 (in vivo t 1 / 2 220 minutes). Even at a higher dose (3 mg / kg or about 3 μmol / kg), 42 was completely cleared from plasma at 4 hours (Table VII), while 43 is expected to drop less than 3 half-lives at 6 hours and maintain sufficient concentrations to produce observable effects. Therefore, a longer in vivo half-life achieves a longer cardiac effect, especially for the macrocyclic 43.

[0464] The scope of the claims should not be limited by the embodiments listed in the examples, but should be given the broadest interpretation consistent with the description as a whole.

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Claims

1. A compound of formula (II): in: X1 does not exist or is X7-X8, where X7 is -(CH2)q-CH3 or -(CF2)q-CF3, wherein q is 0 to 11, a natural amino acid, a synthetic amino acid, the side chain of the natural amino acid, the synthetic amino acid is H, -(C1-C12)alkyl, -(CF2)q-CF3, wherein q is 0 to 11, -(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -(CH2)p-(C3-C 8) cycloalkyl or -(CH2)p-(C3-C8)heterocycloalkyl, wherein p is 0 to 5, wherein the cycloalkyl, the heterocycloalkyl, the aryl or the heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the alkyl, the heteroaryl, the aryl, the cycloalkyl and the heterocycloalkyl are optionally substituted with one or more substituents, wherein Each substituent is independently, for example, halogen, amine, -OH, S, -(C1-C6)alkyl, -O-(C1-C6) alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, and wherein the heteroatom in the heteroalkyl, heteroaryl or heterocycloalkyl is N, O or S; and X8 is absent or is a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8) Cycloalkyl or -(C3-C8)heterocycloalkyl is fused; and wherein the heteroatoms in the heteroalkyl, the heteroaryl or the heterocycloalkyl are 1, 2 or 3 N, O or S; Y is absent or is NH2-, Ac-NH-, guanidine or H; A is -(CH2)n-; -(CH2)nNH=C(NH2)N-CH2-CH=CH- (preferably allyl-glycine or Nα-allyl-arginine), where n is 2, 3 or 4; or -CH=CH-(CH2)m-, where m is 0, 1 or 2; B does not exist or wherein R is O, P, m-alkyl, halogen or nitro, and n is 1, 2 or 3; wherein R is H, C3-C7 alkyl, benzyl or arylalkyl, and n is 1, 2 or 3; wherein n is 1, 2, 3 or 4, and m is 0 or 1; or wherein X9 is CH or N; X2 and X3 are each independently absent or a natural or synthetic amino acid whose side chain is -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl or -(CH2)p-(C3-C8)heteroaryl, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted with at least one amino or guanidine group; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S; X4 is a natural or unnatural amino acid with a positively charged or uncharged side chain; X5 is Gly, Phe, Leu, He, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn); and X6 is X 10 -X 11 -X 12 ,in X 10 is any natural amino acid; or a synthetic amino acid, wherein the side chain of the natural amino acid or the synthetic amino acid is H, -(CH2)p-(C3-C8)alkyl, -(CH2)p-(C3-C8)heteroalkyl, -(CH2)p-(C3-C8)cycloalkyl, -(CH2)p-(C3-C8)heterocycloalkyl, -(CH2)p-(C3-C8)aryl, -(CH2)p-(C3-C8)heteroaryl, -CH2-(CH2)p-NH 2、 -CH2-(CH2)p-guanidine, wherein p is 0 to 5, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with one or more substituents, wherein each substituent is independently, for example, halogen, amino, guanidino, -OH, S, -(C1-C6)alkyl, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p' is 0 to 5; wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally fused with one or two (C3-C8)aryl, (C3-C8)heteroaryl, (C3-C8)cycloalkyl or -(C3-C8)heterocycloalkyl; and wherein the heteroatoms in the heteroalkyl, heteroaryl or heterocycloalkyl are 1, 2 or 3 N, O or S. In a specific embodiment, X 10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently O-(C1-C6)alkyl, -(CH2)p-(C3-C8)aryl, -O-(CH2)p-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p is 0 to 5. In a specific embodiment, X10 is not Ala. In a more specific embodiment, X 10 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Phe, Tic ((S)-N-Fmoc-tetrahydroisoquinoline-3-carboxylic acid), Tyr, 1Nal, 2Nal, TyrOBn, cypTyr(OBn), dcypTyr(OBn), cypTyr(OCyp), cypTyr(OPr), D-1Nal, D-2Nal, D-TyrOBn, or D-Tyr; X 11 is absent or is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn). In one embodiment, X11 is absent or is Pro; and X 12 Does not exist or is Phe, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

2. The compound according to claim 1, wherein: -X2 and X3 are each independently an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4; and / or -X 10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently -OH, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p' is 0 to 5, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

3. The compound according to claim 1, wherein: - X2 and X3 are each independently Lys, Orn, Dab (2,4-diaminobutyric acid), Dap (2,3-diaminopropionic acid), Arg, hArg, His, Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine) or α-methylphenylalanine; -X4 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn); - X5 is Gly, Phe, Leu, Ile, Ser, Aib, Pro, Sar, Oic, βAla, Hyp or Hyp(OBn); and / or -X 10 Yes X 10 is Nle, α-methylleucine, cycloleucine, tert-leucine, cyclohexylalanine (e.g., 3-cyclohexyl-L-alanine), α-methylphenylalanine, Phe, Tic ((S)-N-Fmoc-tetrahydroisoquinoline-3-carboxylic acid), Tyr, 1Nal, 2Nal, TyrOBn, cypTyr(OBn), dcypTyr(OBn), cypTyr(OCyp), cypTyr(OPr), D-1Nal, D-2Nal, D-TyrOBn or D-Tyr, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

4. The compound according to claim 3, wherein: - X2 and X3 are each independently Lys, Arg, hArg, Nle, Leu, Phe or Cha; - X4 is Gly; and / or -X5 is Pro, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

5. The compound according to any one of claims 1 to 4, wherein: -X1 does not exist; -Y is NH2, Ac-NH-, guanidine or H; -A is -CH=CH-(CH2)m-, wherein m is 0, 1 or 2; -B does not exist or wherein R is O, P, m-alkyl, halogen or nitro, and n is 1, 2 or 3, wherein X9 is CH or N; and / or -X 10 is an amino acid whose side chain is -(CH2)p-(C3-C8)alkyl or -(CH2)p-(C3-C8)aryl, wherein p is 0 to 5, wherein the aryl is optionally fused to one or two (C3-C8)aryl groups, and wherein the aryl is optionally substituted with one or more substituents, wherein each substituent is independently -OH, -O-(C1-C6)alkyl, -(CH2)p'-(C3-C8)aryl, -O-(CH2)p'-(C3-C8)aryl, -(C3-C8)cycloalkyl or -O-(C3-C8)cycloalkyl, wherein p' is 0 to 5, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

6. The compound according to claim 5, wherein X 10 It is Nle or D-1Nal or a stereoisomer or a mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

7. A compound according to any one of claims 1 to 4, wherein: - X1 is X7-X8; and / or -Y does not exist, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

8. The compound according to claim 7, wherein: -X1 is X7-X8, and X8 is an amino acid whose side chain is -CH2-(CH2)p-guanidine, -CH2-(CH2)p-NH2 or -(CH2)p-imidazole, preferably -CH2-(CH2)p-guanidine or -CH2-(CH2)p-NH2, wherein p is 0 to 4, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

9. The compound according to claim 7, wherein -A is -(CH2)n- or -CH=CH-(CH2)m-, wherein m is 0, 1 or 2, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

10. A compound of any one of formulae (I) to (VIII) or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

11. The compound according to claim 10, which is any one of compounds 3-4, 9-29, 35-46, 62-70, 72-79, 84 and 89-94, preferably any one of compounds 11, 13, 15-16, 18-20 and 42-44: wherein X represents allylglycine and dX represents D-allylglycine, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

12. The compound according to claim 11, which is any one of compounds 13-25, 27-29, 35-37 and 42-45, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

13. A pharmaceutical composition comprising a compound, stereoisomer, mixture, pharmaceutically acceptable salt, ester or solvate according to any one of claims 1 to 12, and at least one pharmaceutically acceptable carrier or excipient.

14. Use of an effective amount of a compound of any one of formula (I) to (IV) or a stereoisomer or mixture thereof or a pharmaceutically acceptable salt, ester or solvate thereof in the preparation of a medicament for treating a cardiovascular disease in a subject in need thereof.

15. The use according to claim 14, wherein the compound is any one of compounds 3-4, 9-29 and 35-46 as defined in claim 10, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

16. Use according to claim 14, wherein the compound has formula (II) as defined in any one of claims 1 to 8.

17. The use according to claim 16, wherein the compound is any one of compounds 13-25, 27-29, 35, 36-37 and 42-45 as defined in claim 10, preferably any one of compounds 13, 15-16, 18-20, 23 and 42-44, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

18. The use according to claim 17, wherein the compound is compound 42 or 43, or a stereoisomer or mixture thereof, or a pharmaceutically acceptable salt, ester or solvate thereof.

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