PCSK9 antagonist compounds

Abstract

To provide antagonists of PCSK9.SOLUTION: Provided are compounds having a particular structure or salts thereof. The compounds have properties for antagonizing PCSK9.Also described are pharmaceutical formulations comprising the compounds of formula I or their salts, and methods for treating cardiovascular diseases and conditions related to PCSK9 activity, e.g. atherosclerosis, hypercholesterolemia, coronary heart disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular disease and cardiometabolic conditions.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. patent application Ser. No. 62 / 687,913, filed June 21, 2018. This application claims priority to US Pat. No. 6,399,433, which is incorporated herein by reference in its entirety. Can be enjoyed. [Background technology]

[0002] The identification of compounds and / or agents effective in the treatment of cardiovascular disease is highly desirable. In clinical trials, lowering LDL cholesterol levels has been shown to reduce coronary events. Law et al., 2003 BMJ 326:1423-14 27. Moderate reductions in plasma LDL cholesterol levels over a lifetime are associated with reduced risk of coronary events. was found to correlate with a substantial reduction in the incidence of NE ngl.J.Med.354:1264-1272. This is a non-lipid-related cardiovascular risk This was also the case in populations with high prevalence of the factor; see above. The benefits of managing terror level control are great.

[0003] Proprotein convertase subtilisin-kexin type 9 (hereinafter referred to as "PCSK9") Also known as neuronal apoptosis-regulating converting enzyme ("NARC-1"), A proteinase K-like subtilase identified as the ninth member of the subtilase family. See Seidah et al., 2003 PNAS 100:928-933. PCSK9 is a member of the mammalian proprotein convertase family of serine proteases. and contains an N-terminal signal sequence, a prodomain, a catalytic domain, and a C-terminal domain. Seidah et al., 2012 Nat. Rev. Drug Discov. 11:3 As seen in other genes involved in cholesterol metabolism, In addition, studies of PCSK9 transcriptional regulation have demonstrated that it is a sterol regulatory element-binding protein ("PCSK9"). have demonstrated that it is regulated by SREBP (Maxwell et al., 2003). J. Lipid Res. 44:2109-2119, which has implications for lipoprotein metabolism. Typical of other genes involved; Dubuc et al., 2004 Arterioscler .Thromb.Vasc.Biol.24:1454-1459. Statins are a class of drugs Upregulating PCSK9 expression in a manner that results in a cholesterol-lowering effect Moreover, the PCSK9 promoter is involved in cholesterol regulation. It has been shown to contain two conserved sites involved: a sterol regulatory element and an Sp1 site. has been mentioned above.

[0004] While present in the endoplasmic reticulum, PCSK9 binds Gln-152 and Autocleavage between Ser-153; Naureck iene et al., 2003 Arch.Biochem.Biophys.420:55-6 7;Seidah et al., 2003 Proc. Natl. Acad. Sci. USA 100:928-933. Subsequent trafficking through the trans-Golgi network During cleavage, the prodomain remains in close association with the catalytic domain. It has been demonstrated that maturation is important for the secretion and subsequent extracellular function of PCSK9. (Benjannet et al., 2012 J. Biol. Chem. 287:337 (See, e.g., 45-33755). Thus, several lines of evidence support the role of PCSK9 in It reduces the amount of hepatic LDLR protein, thus removing LDL cholesterol from circulation. Studies have demonstrated that nicotine impairs the liver's ability to clear it from

[0005] Adenovirus-mediated overexpression of PCSK9 in mouse liver increases hepatic LDLR expression The dramatic loss of protein leads to the accumulation of circulating LDL-C and increases LDLR mRNA levels. Benjannet et al., 2004 J. Biol. Chem. 279 :48865-48875;Maxwell&Breslow,2004 PNAS 1 01:7100-7105;Park et al., 2004 J. Biol. Chem. 279: 50630-50638; and Lalanne et al., 2005 J. Lipid Res. 46:1312-1319. PCA response to elevated circulating LDL-C levels in mice. The effect of SK9 overexpression was entirely dependent on expression of the LDLR, which also regulates PCSK9. The regulation of LDL-C by IL-1 is mediated by downregulation of LDLR protein. Consistent with these findings, mice lacking PCSK9 or Mice in which PCSK9 mRNA was reduced by a sense oligonucleotide inhibitor had liver High levels of LDLR protein and a better ability to clear circulating LDL-C; Ashid et al., 2005 PNAS 102:5374-5379; and Graham et al., 2007 J. Lipid Res. 48(4):763-767. In addition, Reducing PCSK9 levels in mouse hepatocytes by siRNA also resulted in higher Ben Jannet et al., 2004 J.Biol.Chem.279:48865-48875 and Lalanne et al., 2005 J. Lipid Res. 46:1312-13 19 Taken together, these data suggest that the action of PCSK9 is mediated by upregulation of LDLR protein levels. These results indicate that lowering cholesterol leads to an increase in LDL-C.

[0006] Many mutations in the PCSK9 gene also cause autosomal dominant hypercholesterolemia (" The disease is critically linked to the increased levels of low-density lipoproteins ("LDL") in the plasma. A genetic metabolic disorder characterized by a marked increase in "DL" particles, which leads to premature cardiovascular failure. Abifadel et al., 2003 Nature Genetics 3 4:154-156; Timms et al., 2004 Hum. Genet. 114:349- 353; see Leren, 2004 Clin. Genet. 65:419-422. The later published study by Abifadel et al. on the S127R mutation Patients carrying such mutations (1) have apoB100-containing lipoproteins, e.g., low-density lipoproteins Lipoproteins ("LDL"), very low density lipoproteins ("VLDL") and intermediate density lipoproteins ("VLDL"). (2) overproduction of intracellular lipoproteins ("IDLs") and the like, and Higher plasma total cholesterol and cholesterol due to the concomitant reduction in clearance or conversion. reported that the expression of apoB100 was increased in patients with rheumatoid arthritis; Ouguerram et al., 2004 Ar terioscler.Thromb.Vasc.Biol.24:1448-1453 .

[0007] Thus, there is no doubt that PCSK9 plays a role in regulating LDL. PCSK9 expression or upregulation is not associated with vascular LDL cholesterol levels. Corresponding inhibition or loss of expression of PCSK9 is associated with increased plasma levels of LDL cholesterol. PCSK9 sequence variants have been associated with reduced plasma levels of steroids. Reducing cholesterol levels has been found to confer protection against coronary heart disease. Cohen, 2006 N. Engl. J. Med. 354:1264-1272.

[0008] Thus, there are numerous potential applications of PCSK9 in cardiovascular disease, including antagonizing the role of PCSK9 in LDL regulation. The identification of compounds and / or agents effective in the treatment of gastrointestinal disorders is highly desirable. Generally, PCSK9 circulates in the blood and binds to LDL receptors on the cell surface with high affinity. Since the level of LDL in serum is not high, this mechanism may be of use in the treatment of diseases associated with high serum LDL levels. Previous attempts to exploit this phenomenon have focused on the use of biopolymers such as antibodies. Therefore, studies using short peptides or small molecules to inhibit PCSK9 have been proposed. There are only a few publications reflecting activity against this target. For example, Zhang et al., 2014 See J. Biol. Chemistry, 289(2):942-955. Moreover, the oral route of administration of such compounds is expected to play a role in modulating PCSK9 activity. This is a highly desirable route to providing treatment for symptoms that can be effectively treated. There are a small number of compounds that are amenable to formulation into dosage forms for utilizing this route. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Law et al., 2003 BMJ 326:1423-1427 [Non-Patent Document 2] Cohen et al., 2006 N. Engl. J. Med. 354:1264-1272 [Non-Patent Document 3] Seidah et al., 2003 Proc. Natl. Acad. Sci. USA 100:928-933 [Non-Patent Document 4] Seidah et al., 2012 Nat. Rev. Drug Discov. 11:367-383 [Non-Patent Document 5] Maxwell et al., 2003 J. Lipid Res. 44:2109-2119 [Non-Patent Document 6] Dubuc et al., 2004 Arterioscler.Thromb.Vasc.Biol.24:1454-1459 [Non-Patent Document 7] Naureckiene et al., 2003 Arch.Biochem.Biophys.420:55-67 [Non-Patent Document 8] Benjannet et al., 2012 J. Biol. Chem. 287:33745-33755 [Non-Patent Document 9] Benjannet et al., 2004 J. Biol. Chem. 279:48865-48875 [Non-Patent Document 10] Maxwell&Breslow,2004 PNAS 101:7100-7105 [Non-Patent Document 11] Park et al., 2004 J. Biol. Chem. 279:50630-50638 [Non-Patent Document 12] Lalanne et al., 2005 J.Lipid Res.46:1312-1319 [Non-Patent Document 13] Rashid et al., 2005 PNAS 102:5374-5379 [Non-Patent Document 14] Graham et al., 2007 J. Lipid Res. 48(4):763-767 [Non-Patent Document 15] Lalanne et al., 2005 J.Lipid Res.46:1312-1319 [Non-Patent Document 16] Abifadel et al., 2003 Nature Genetics 34:154-156. [Non-Patent Document 17] Timms et al., 2004 Hum. Genet. 114:349-353 [Non-Patent Document 18] ;Leren,2004 Clin.Genet.65:419-422 [Non-Patent Document 19] Ouguerram et al., 2004 Arterioscler.Thromb.Vasc.Biol.24:1448-1453 [Non-Patent Document 20] Zhang et al., 2014 J.Biol.Chemistry,289(2):942-955 Summary of the Invention [Means for solving the problem]

[0010] The present invention inhibits the activity of PCSK9 and provides a therapeutic effect by administering a PCSK9 antagonist. These drugs could be used to inhibit the corresponding role that PCSK9 plays in a variety of conditions, including: The present study advances these interests by providing a novel PCSK9 antagonist that can That is why.

[0011] In one embodiment, the present invention provides a compound of formula I: [ka]

[0012] A compound of the formula: X is H, F, Cl or Br; R 1 teeth: (a)-H; or (b)-(CH2) z -R 14A wherein z is 1 to 6; R 14A teeth: (i)-H; (ii) -NH2; (iii)-N + H3; (iv)-N + (H3C)3; (v)-NH-C(O)-[(CH2)2-O-]2-(CH2)2R 14B (In the formula, R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH3)3 be); (vi) -NH-C(O)-[(CH y12 -O-]2-(CH2) y13 R 14 B (Wherein: y12 and y13 are not both 2 at the same time, and are independently 2 to 4. ; and R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (C H3)3); (vii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; R 14C is -O-(CH2) za -N + (CH3)3, where za is 3 or is 4); and (viii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; , R 14C teeth: (ai)-O-(CH2)2-N + (CH3)3; (aii)-N + (CH3)3; or Formula (aiii): [ka]

[0013] (part of); R 2 teeth: (a)-H; and (b)-(CH2) z -R 14A wherein z is 1 to 6; R 14A teeth: (i)-H; (ii) -NH2; (iii)-N + H3; (iv)-N + (H3C)3; (v)-NH-C(O)-[(CH2)2-O-]2-(CH2)2R 14B (In the formula, R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH3)3 be); (vi) -NH-C(O)-[(CH y12 -O-]2-(CH2) y13 R 14 B (Wherein: y12 and y13 are not both 2 at the same time, and are independently 2 to 4. ; and R 14B is: -NH2;-N + H3;-N(CH3)2; or -N+ (C H3)3); (vii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; R 14C is -O-(CH2) zb -N + (CH3)3, where zb is 3 or is 4); and (viii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; , R 14C teeth: (ai)-O-(CH2)2-N + (CH3)3; (aii)-N + (CH3)2R 14ca (In the formula, R 14ca is -CH3 or -(CH2) 1-4 -OCH3); Formula (aiii): [ka]

[0014] or (aiv) formula: [ka]

[0015] (wherein R 14Cb and R 14Cc is selected from 1 to 4); or R 1 and R 2 can be combined together to form the formula: [ka]

[0016] may form a portion of the formula: G 1 , R G1a and R G1b is defined as follows: (a)G 1 is the expression: [ka]

[0017] wherein n q1 is 1 to 6, m q1 is 0, 1 or 2 In total, n q1 and m q1 The values ​​of are defined as the length of the linker portion that they define. Carbon atoms in the chain, including the carbon atom in the chain that forms the carbonyl moiety, and / or The total number of oxygen atoms is selected to be not more than eight; R G1a is selected from: (i) -H; and (ii) alkyl of up to 4 carbon atoms; and R G1b teeth: (i) Formula: [ka]

[0018] Part of; and (ii) Formula: [ka]

[0019] or (b) G 1 is the expression: [ka]

[0020] is a linker moiety,q2 is 0, 1 or 2, m q2 is 1 to 6 Yes, in total n q2 and m q2 The values ​​of are the lengths of the linker parts they define. and / or selected to contain not more than 8 oxygen atoms in total; R G1a teeth: (i) Formula: [ka]

[0021] Part of; and (ii) Formula: [ka]

[0022] and R G1b is selected from: (i) -H; and (ii) alkyl of up to 4 carbon atoms; R 8 is -CH3 or the formula: [ka]

[0023] where R 8a is -H or a straight, branched or cyclic 4 is alkyl of up to 10 carbon atoms; A is: (a) Formula: [ka]

[0024] Part of; (b) -CH2-(CH2) y-CH2- (wherein y is 1 to 6); (c) Formula: [ka]

[0025] (wherein A b1 teeth: (i) Formula: [ka]

[0026] where x is 1 to 6; or (ii) Formula: [ka]

[0027] where y is 1 to 5; (d) Formula: -CH2-(CH2) m -O-(CH2) n - portion (wherein m is 1 to 5 and n is 0 or 1 to 4. Selected from; B is: (a) combination; (b)-(CH2) 1-4 ;or (c) Formula: [ka]

[0028] is part of; D is: (a) Formula: [ka]

[0029] (wherein E is -CH2- or -(CH2) 2-4 -O-, and A and B are , as defined above); Formula (b): [ka]

[0030] wherein A and B are as defined above; (c) Formula: [ka]

[0031] (wherein, n a is 1, 2 or 3, m a is 2, 3, or 4, and n a + m a is ≧3, where A and B are as defined above; (d) Formula: [ka]

[0032] (wherein R 34b is -H or a straight, branched or cyclic group having up to four and A and B are as defined above. or a pharma- ceutically acceptable salt of any of them.

[0033] In a further embodiment, the present invention relates to a compound of formula I, wherein X is F. The compound or any pharma- ceutically acceptable salt thereof is provided. In the embodiment, D is a group represented by the formula: [ka]

[0034] wherein E is -CH2- or -(CH2)2-O-. where A and B are as defined herein.

[0035] In some embodiments, D is of the formula: [ka]

[0036] wherein E is -CH2- or -(CH2)2-O-. where A and B are as defined herein.

[0037] In some embodiments, D is of the formula: [ka]

[0038] wherein A and B are as defined herein. do.

[0039] In some embodiments, D is of the formula: [ka]

[0040] wherein A and B are as defined herein. do.

[0041] In some embodiments, D is of the formula: [ka]

[0042] wherein A and B are as defined herein. do.

[0043] In some embodiments, D is of the formula: [ka]

[0044] wherein A and B are as defined herein. do.

[0045] In some embodiments, D is of the formula: [ka]

[0046] wherein A and B are as defined herein. do.

[0047] In some embodiments, D is of the formula: [ka]

[0048] wherein A and B are as defined herein. do.

[0049] R 1 and R 2 join together with the peptide ring to which they are attached to form a ring structure In some embodiments, R 1 and R 2 The structure: [ka]

[0050] It is preferred to form part of

[0051] In one embodiment, the present invention provides a compound of the present invention, e.g., a compound of formula I, Pharmaceutical compositions, preferably intended for oral administration, containing at least one pharmaceutical excipient to provide.

[0052] In one embodiment, the present invention provides a method for administering to a subject in need thereof a therapeutically effective amount of a compound of formula I or or a salt thereof, preferably in the form of a pharmaceutical composition, to inhibit PCSK9 activity. Related disease conditions, e.g., atherosclerosis, hypercholesterolemia, coronary heart disease Disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular and cardiovascular diseases The present invention provides a method of antagonizing PCSK9 in providing treatment for neurological conditions. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0053] In the following description, conventional structural representations are used, which are The invention includes conventional stereochemical notations for the following:

[0054] Therefore, the structural representation of the compounds of the present invention includes some asymmetric carbon atoms shown in the exemplary compounds. The conventional stereochemical notation for the chiral centers is included. In this case, solid black "wedge" bonds represent bonds that protrude from the plane of the replication medium, and "hatch" bonds represent bonds that protrude from the plane of the replication medium. The "hashed" wedge bond represents a downward bond to the plane of the replication medium, and double The "wavy" line attached to the carbon bearing the bond indicates that both cis and trans orientations are possible. As is conventional, solid lines indicate inclusion of Therefore, if no specific stereochemical designation is given, When used herein, the representation contemplates all stereochemical and spatial orientations of structural features.

[0055] As shown in the examples of the present invention and as mentioned above, certain asymmetric carbon centers The structure is represented using the conventional "solid wedge" and "hash wedge" bond representations. For the most part, the absolute configuration of the exemplified compounds has not been determined, but the Prepared using the same or similar reaction conditions and starting reagents and under the same chromatographic conditions. The stereochemical configuration (determined by X-ray crystallography) of the compound is known. The structural elements are assigned by analogy to certain exemplary compounds. The specific assignment of configuration depicted in the figure indicates that the specific compound prepared is one specific This means confirming that the compound has an excess of stereoisomers, and particular attention should be paid to the data presented. Unless otherwise stated, nothing in this specification necessarily provides an absolute determination of the stereochemical configuration of said compounds. It is not intended as a statement of the invention.

[0056] When a mixture of isomers is obtained, the individual stereoisomers are separated in high enantiomeric excess percentages. Preparation in situ can be accomplished, if desired, by separation of the mixture using conventional methods, e.g. For example, by chromatography or crystallization, or for the synthesis described, stereochemistry may be used. This can be achieved by using substantially uniform starting materials or by stereoselective synthesis. It is understood that derivatization may be performed prior to separation of stereoisomers. The separation of the mixture can be carried out at an intermediate step in the synthesis of the compounds of formula I or as a final step. This can be done on racemic products.

[0057] Where indicated herein, absolute stereochemistry may be determined from X-ray crystallography of crystalline products or crystalline intermediates. analytically determined, if necessary, by the addition of a reagent containing an asymmetric center of known configuration. The specific heterocycles of such racemates, enantiomers or diastereomers are derivatized with Unless the form, salt, solvate (including hydrate) or solvated salt is indicated, The disclosure includes all such isomers, as well as such racemates, enantiomers, diastereomers, and mixtures thereof. and mixtures thereof, including salts, solvates (including hydrates) and solvated salts.

[0058] The present invention also includes isotopically labeled compounds of the present invention, which are described herein. Structurally identical to the compounds listed, but with a statistically significant percentage of that form of the compound. One or more atoms in the number are equal to the atomic mass or mass of the most abundant isotope commonly found in nature. is replaced by an atom with a different atomic mass or mass number than the mass number, In addition, the natural abundance of that isotope present in the compounds of the present invention varies. is meant to encompass all suitable isotopic variations of the compounds of Formula I.

[0059] Examples of isotopes that may be preferentially incorporated into the compounds of the present invention include hydrogen, carbon, Isotopes of nitrogen, oxygen, phosphorus, iodine, fluorine and chlorine, including but not limited to Iga: 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O , 31 P, 32 P, 35 S, 18 F and 36 Cl, 123 I and125 Including I. Other It will be understood that isotopes of can also be incorporated by known means.

[0060] In particular, certain isotopically labeled compounds of the invention (e.g., 3 H, 11 C and 14 C) can be synthesized by combining compounds and / or substrates using a variety of known techniques. It is recognized that these are particularly useful in tissue distribution assays. The compound of the present invention is protium ( 1 H) and deuterium ( 2 H or D) Protium is the predominant hydrogen isotope found in nature. Enrichment in deuterium may provide certain therapeutic advantages, such as increased in vivo half-life or may provide reduced dosage requirements or may be useful as standards for characterization of biological samples; Isotopically enriched compounds within the scope of Formula I can be prepared by conventional methods well known to those skilled in the art. by conventional techniques or as described in the schemes and examples herein. By a process similar to that described above, excess isotope can be produced using appropriate isotopically enriched reagents and / or intermediates. can be prepared without much experimentation.

[0061] If the wavy line is at the end of a conventional bond (as opposed to connecting two atoms within a structure), Generally, this indicates the point of attachment to the structure, for example: [ka]

[0062] is a secondary butyl moiety attached via a methylene group through a bond that ends in a wavy line. When alphabetic notations are used to represent substituent moieties, dashes indicate The symbol -CH2-C is used to indicate the point of attachment to the substrate shown, e.g. -CH2-C (O)-CH2Cl is an acetyl chloride moiety that is bonded via the methylene moiety of this moiety. This indicates that.

[0063] Any variable group (e.g., n, R a , R b etc.) in any component or in formula I When it occurs more than once in, the following applies for each occurrence, unless otherwise specified at the time of definition: Its definition in any one occurrence is independent of its definition in any other occurrence. Present, i.e. R 1 , R A The selection of the various combinations of substituents defined in The substituents and substituent groups should be selected according to well-known principles of structural connectivity and stability. and / or combinations of variables are acceptable if such combinations result in stable compounds. Recognize that only certain actions are acceptable.

[0064] A "stable" compound is one that can be prepared and isolated, and whose structure and properties are Use of the compounds for the purposes described herein (e.g., therapeutic administration to a subject) Remain essentially unchanged or to remain unchanged for a period sufficient to permit The compounds of the present invention are not limited to stable compounds encompassed by Formula I. It is determined.

[0065] Any variable group or moiety may be in the form of a range, e.g., (-CH 2- ) 1-4 In the case expressed as If the range is 1, then both ends of the specified range (i.e., 1 and 4 in this example) are inclusive, and similarly All whole numbers in between (ie, 2 and 3 in this example) are included.

[0066] The term "halogen" includes fluorine, chlorine, bromine and Contains iodine.

[0067] As used herein, a "subject" (or "patient") is In need of treatment, animals, preferably mammals, especially humans, or livestock animals and domestic animals "Animal Crossing" refers to non-human animals, including livestock, including, but not limited to, cattle. , horses, sheep, pigs, goats, rabbits, cats, dogs, and other mammals. In some embodiments, the subject is preferably a human. The term "administration" and variations thereof with respect to compounds of Formula I (e.g., "administering" a compound) are intended to be used in the present invention. and " ) provides a compound or a pharma- ceutically acceptable salt thereof to a subject in need of treatment. This means that...

[0068] As mentioned above, in one embodiment, the present invention provides a compound of formula I or a pharma- ceutically acceptable salt thereof. The present invention also includes providing acceptable salts thereof, which have the property of antagonizing the function of PCSK9.

[0069] In one embodiment, the compound of formula I has formula IA: [ka]

[0070] wherein: R 1 teeth: (a)-H; or (b)-(CH2) z -R 14A wherein z is 1 to 6; R14A teeth: (i)-H; (ii) -NH2; (iii)-N + H3; (iv)-N + (H3C)3; (v)-NH-C(O)-[(CH2)2-O-]2-(CH2)2R 14B (In the formula, R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH3)3 be); (vi) -NH-C(O)-[(CH y12 -O-] 1-4 -(CH2) y13 R 14B , preferably -NH-C(O)-[(CH2) y12 -O-]2-(CH2) y1 3R 14B (Wherein: y12 and y13 are not both 2 at the same time, and are independently 2 to 4 and R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH3)3); (vii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; R 14C is -O-(CH2) za -N + (CH3)3, where za is 3 or is 4); and (viii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; , R 14C teeth: (ai)-O-(CH2)2-N + (CH3)3; (aii)-N + (CH3)3; or Formula (aiii): [ka]

[0071] (part of); R 2 teeth: (a)-H; and (b)-(CH2) z -R 14A wherein z is 1 to 6; R 14A teeth: (i)-H; (ii) -NH2; (iii)-N + H3; (iv)-N + (H3C)3; (v) -NH-C(O)-[(CH2)2-O-] 1-4 -(CH2)2R 14B , good Preferably -NH-C(O)-[(CH2)2-O-]2-(CH2)2R 14B (In the formula, R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH3)3 be); (vi) -NH-C(O)-[(CH y12 -O-]2-(CH2) y13 R 14 B (Wherein: y12 and y13 are not both 2 at the same time, and are independently 2 to 4. ; and R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (C H3)3); (vii) -NH-C(O)-(CH y R 14C(wherein y is 1 to 6; R 14C is -O-(CH2) zb -N + (CH3)3, where zb is 3 or is 4); and (viii) -NH-C(O)-(CH y R 14C (wherein y is 1 to 6; , R 14C teeth: (ai)-O-(CH2)2-N + (CH3)3; (aii)-N + (CH3)2R 14ca (In the formula, R 14ca is -CH3 or -(CH2) 1-4 -OCH3); Formula (aiii): [ka]

[0072] or (aiv) formula: [ka]

[0073] (wherein Y 14Cb and Y 14Cc is selected from 1 to 4); and teeth R 1 and R 2 can be combined together to form the formula: [ka]

[0074] may form a portion of the formula: G 1 , R G1a and R G1b is defined as follows: (a)G 1 is the expression: [ka]

[0075] wherein n q1 is 1 to 6, m q1 is 0, 1 or 2 In total, n q1 and m q1 The values ​​of are defined as the length of the linker portion that they define. Carbon atoms and / or carbon atoms that make up the chain, including the carbon atom in the chain that forms the carbonyl moiety are selected so that the total number of oxygen atoms does not exceed 8; R G1a is selected from: (i) -H; and (ii) alkyl of up to 4 carbon atoms; and R G1b teeth: (i) Formula: [ka]

[0076] Part of; and (ii) Formula: [ka]

[0077] or (b) G 1 is the expression: [ka]

[0078] wherein n q2 is 0, 1 or 2, m q2 is 1 to 6 In total, n q2 and m q2The values ​​of are defined as the length of the linker portion that they define. Carbon atoms and / or carbon atoms that make up the chain, including the carbon atom in the chain that forms the carbonyl moiety are selected so that the total number of oxygen atoms does not exceed 8; R G1a teeth: (i) Formula: [ka]

[0079] Part of; and (ii) Formula: [ka]

[0080] and R G1b is selected from: (i) -H; and (ii) alkyl of up to 4 carbon atoms; R 8 is -CH3 or the formula: [ka]

[0081] where R 8a is -H or a straight, branched or cyclic 4 is alkyl of up to 10 carbon atoms; A is: (a) Formula: [ka]

[0082] Part of; (b) -CH2-(CH2) y -CH2- (wherein y is 1 to 6); Formula (c): [ka]

[0083] (wherein A b1 teeth: (i) Formula: [ka]

[0084] where x is 1 to 6; or (ii) Formula: [ka]

[0085] where y is 1 to 5; (d) Formula: -CH2-(CH2) m -O-(CH2) n - portion (wherein m is 1 to 5 and n is 0 or 1 to 4. Selected from; B is: (a) combination; (b)-(CH2) 1-4 ;or (c) Formula: [ka]

[0086] is part of; D is: (a) Formula: [ka]

[0087] (wherein E is -CH2- or -(CH2) 2-4 -O-, and A and B are , as defined above); Formula (b): [ka]

[0088] wherein A and B are as defined above; (c) Formula: [ka]

[0089] (wherein, n a is 1, 2 or 3, m a is 2, 3, or 4, and n a + m a is at least 3, where A and B are as defined above; (d) Formula: [ka]

[0090] (wherein R 34b is -H or a straight, branched or cyclic group having up to four and A and B are as defined above. or a pharma- ceutically acceptable salt of any of them.

[0091] In an embodiment of the compound of formula IA, R 1 is -(CH2) z -R 14A where : z is 1 to 6, R 14A teeth: (i)-H; (ii) -NH2; (iii)-N + H3; or (iv)-N + (H3C)3; R 2 is -(CH2) z -R 14A where z is 1 to 6; R 14A teeth: (i)-H; (ii) -NH2; (iii) -NH-C(O)-[(CH2)2-O-] 1-4 -(CH2)2R 14B , Preferably, -NH-C(O)-[(CH2)2-O-]2-(CH2)2R 14B (In the ceremony , R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH3)3 is); (iv) -NH-C(O)-[(CH y12 -O-]2-(CH2) y13 R 14B (Wherein: y12 and y13 are independently 2 to 4, but not both simultaneously 2; And R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + (CH 3) 3); (v) -NH-C(O)-(CH2) y R 14C (wherein y is 1 to 6; R 14 C is -O-(CH2) zb -N + (CH3)3, where zb is 3 or 4. ); and (vi) -NH-C(O)-(CH2) y R 14C (wherein y is 1 to 6; R 1 4C teeth: (ai)-O-(CH2)2-N + (CH3)3; (aii)-N + (CH3)2R 14ca (In the formula, R 14ca is -CH3 or - (CH2) 1-4 -OCH3); Formula (aiii): [ka]

[0092] or (aiv) formula: [ka]

[0093] (wherein Y 14Cb and Y 14Cc is selected from 1 to 4); and teeth R 8 is -CH3 or the formula: [ka]

[0094] where R 8a is -H or a straight, branched or cyclic 4 is alkyl of up to 10 carbon atoms; A is: (a) Formula: [ka]

[0095] Part of; (b) -CH2-(CH2) y -CH2- (wherein y is 1 to 6); (c) Formula: [ka]

[0096] (wherein A b1 teeth: (i) Formula: [ka]

[0097] where x is 1 to 6; or (ii) Formula: [ka]

[0098] where y is 1 to 5; and (d) Formula: -CH2-(CH2) m -O-(CH2) n - portion (wherein m is 1 to 5 and n is 0 or 1 to 4. Selected from; B is: (a)-(CH2) 1-4 ;or Formula (b): [ka]

[0099] is part of; D is: (a) Formula: [ka]

[0100] (wherein E is -CH2- or -(CH2) 2-4 -O-, and A and B is as defined above); Formula (b): [ka]

[0101] where A and B are as defined above; or Formula (c): [ka]

[0102] (wherein R 34b is -H or a straight, branched or cyclic group having up to four and A and B are as defined above. or any pharma- ceutically acceptable salt thereof.

[0103] In certain embodiments of the compound of formula IA, D is of the formula: [ka]

[0104] wherein E is -CH2- or -(CH2)2-O-, and A and B is as defined above in formula IA.

[0105] In certain embodiments of the compound of formula IA, A is: (a)-(CH2)6; Formula (b): [ka]

[0106] where x is 1 to 3; or Formula (c): [ka]

[0107] This is the part.

[0108] In another embodiment of the compound of formula IA, R 2 teeth: (a)-(CH2) z -R 14A wherein z is 1 to 6; R 14A teeth: (a)-H; (b)-CH3; (c)-NH2; (d)-N + H3; (e)-N + (H3C)3; (f) -NH-C(O)-[(CH2) 2-4 -O-] 2-4 -(CH2) 2-4 R 1 4B (In the formula, R 14B is: -NH2;-N + H3;-N(CH3)2; or -N + ( CH3)3); (g) -NH-C(O)-[(CH2) y R 14C (wherein y is 1 to 6; R 14C teeth: (ai)-O-(CH2) 2-4 -N + (CH3)3; (aii)-N + (CH3)3; or Formula (aiii): [ka]

[0109] (This is part of ;or Formula (b) [ka] This is the part.

[0110] In a further embodiment of the compound of formula IA, R 1 teeth: (a)-H; (b)-(CH2) z -R 14A wherein z is 1 to 6; R 14A teeth: (i)-H; (ii)-N + H3; or (iii)-NH-C(O)-[(CH2)2-O-]2-(CH2)2-N + (CH 3)3 is) is selected from.

[0111] In yet another embodiment of the compound of formula IA, A is -CH2-(CH2) y -CH2 -, where y is 3 to 5. In a further embodiment, A is -(CH 2) The number is 6.

[0112] In another embodiment of the compound of formula IA, B is of the formula: [ka] This is the part.

[0113] In another embodiment of the compound of formula IA, R 1 is -(CH2) z -R 14A And where z is 1 to 6; R 14A is -H. Another embodiment of the compound of formula IA In R 1 is -(CH2) z -R 14A wherein z is 1 and R 14 A is -H.

[0114] In another embodiment of the compound of formula IA, R 2 is -(CH2) z -R 14A And where z is 1 to 6; R 14A is -NH-C(O)-(CH2) y R 14C where y is 1 to 6; R 14C -N + (CH3)2R 14ca Yes In the formula, R 14ca is -CH3.

[0115] In another embodiment of the compound of formula IA, R 8 is the expression: [ka]

[0116] In the formula R 8a is -H or a straight chain of up to 4 carbon atoms In a further embodiment, R 8 is the expression: [ka]

[0117] In the formula R 8b is -H, -CH3, or -C(CH3)3.

[0118] In some embodiments, the compound of formula I has the structure of formula II or formula IIA. or a pharma- ceutically acceptable salt thereof: [ka]

[0119] In the formula, A, R 1 and R 2 is as defined above in formula IA; B 1 Ha-(CH 2) 0-2 and D 1 teeth: a) Formula: [ka]

[0120] Part of; and b) Formula: [ka] is selected from the portion.

[0121] In some embodiments of Formula II or Formula IIA, D 1 is the expression: [ka]

[0122] In some embodiments of Formula II or Formula IIA, D 1 is the expression: [ka]

[0123] In some embodiments of Formula II or Formula IIA, D 1 is the expression: [ka]

[0124] In some embodiments of Formula II or Formula IIA, D 1 is the expression: [ka]

[0125] It is preferable that the part is

[0126] In some embodiments, the compound of formula I is preferably of formula III: [ka]

[0127] wherein A, R1 and R 2 is as defined above in formula IA Ri, D 2 is the expression: [ka] This is the part.

[0128] In some embodiments of Formula III, D 2 is the expression: [ka]

[0129] In some embodiments of formula III, D 2 is the expression: [ka]

[0130] In some embodiments of formula III, D 2 is the expression: [ka] It is preferable that the part is

[0131] In some embodiments, the compound of formula I is preferably of formula IV: [ka]

[0132] wherein A, R 1 and R 2 is as defined above in formula IA do.

[0133] In some embodiments, the compound of formula I has formula V: [ka]

[0134] wherein A, B, R 1 and R 2 is as defined above in formula IA and D 2 teeth: (a) Formula: [ka] Part of;

[0135] Formula (b): [ka] Part of;

[0136] Formula (c): [ka] part of; or

[0137] (d) Formula: [ka] This is the part.

[0138] Some embodiments of Formula I, Formula IA, Formula II, Formula IIA, Formula III, Formula IV, or Formula V In the formula: -(CH2) ya Preferably, ya is a moiety of the formula In some embodiments of Formula I, Formula IA, Formula II, or Formula IIA, A has the formula: -CH2-(CH2) ma -O-(CH2) na - is preferably a part In the formula, ma is 2 or 3, and na is 0 or 1. Formula III, Formula IV and In some embodiments of formula V, A is of the formula: -CH2-(CH2) ma-O-(C H2) na -moiety, where ma is 2 or 4 and na is Formula I, Formula IA, Formula II, Formula IIA, Formula III, Formula IV or Formula V In some embodiments, A is of the formula: [ka]

[0139] wherein yb is 1 to 3. Formula I, Formula IA, Formula II, In some embodiments of formula IIA, formula III, formula IV, or formula V, A is of the formula: [ka] It is preferable that the part is

[0140] In the present specification, the compounds of formula I are represented by compounds Ex-1, Ex-2, Ex-3, E x-4, Ex-5, Ex-6, Ex-7, Ex-8, Ex-9, Ex-10, Ex-11 , Ex-12, Ex-13, Ex-14, Ex-15, Ex-16, Ex-17, Ex- 18, Ex-19, Ex-20, Ex-21, Ex-22, Ex-23, Ex-24, E x-25, Ex-26, Ex-27, Ex-28, Ex-29, Ex-31, Ex-35 , Ex-36, Ex-38, Ex-39, Ex-40, Ex-41, Ex-44, Ex- 47, Ex-48, Ex-49, Ex-50, Ex-51, Ex-52, Ex-53, E x-54, Ex-55, Ex-56, Ex-57, Ex-58, Ex-59, Ex-60 and Ex-61, or any pharma- ceutically acceptable salt thereof. The products are disclosed in Table 1 and are also referred to herein as "compounds of the invention."

[0141] Table 1 [Table 1] TIFF2025041712000093.tif161165TIFF2025041712000094.tif167164TIFF2025041712000095.tif172165TIFF2025041712000096.tif174164 TIFF2025041712000097.tif143165TIFF2025041712000098.tif152164TIFF2025041712000099.tif146165TIFF2025041712000100.tif124168

[0142] In the formula, A - is a pharma- ceutically acceptable anion.

[0143] The terms "salt(s)" and the phrase "pharmaceutically acceptable salt" as used herein Its use in the art includes any of the following: acid salts formed with inorganic and / or organic bases; zwitterionic first salts Quaternary ammonium complexes. Salts of the compounds of the present invention may be formed by methods known to those skilled in the art. For example, a compound of the invention may be reacted with an amount of acid or base, such as an equivalent amount of acid or base. or a base in a solvent, for example in a solvent in which the salt precipitates or in an aqueous solvent, This may then be formed by freeze-drying.

[0144] The compounds of the present invention include a tricoordinate nitrogen atom, e.g., a primary, secondary or tertiary amine. The nitrogen atom of the alkylene moiety is preferably a nitrogen atom, and as is known, the lone pair of electrons present on the nitrogen atom is preferably a Under suitable reaction conditions, it can be protonated with a suitable acid or with a suitable reagent, e.g., an alkyl bromide. This allows the anion generated in the process, e.g., a halogen atom, to be alkylated with A four-coordinate charged nitrogen is provided that is stabilized by an on or conjugate base. The compounds of the invention may be prepared in the form of a free base or may be prepared as a quaternary or salt complex. In some cases, a suitable acidic proton is located proximal to a basic nitrogen. As the term is used herein, the formation of zwitterionic complexes is possible. Salts of the compounds of the present invention may be acid salts formed with inorganic and / or organic acids. Whether basic salts formed with inorganic and / or organic bases, amphoteric ions Salts formed with the inclusion of carboxylic acid properties, for example, when a compound contains a basic moiety (such as but not limited to Nitrogen atoms (e.g. amines, pyridines or imidazoles) and acidic moieties (e.g. The salts may contain both carboxylic acids (such as, but not limited to, quaternary carboxylic acids) and quaternary Any ammonium complexes, whether or not they are ammonium esters, are included within the scope of the compounds of the invention described herein. will be done.

[0145] Thus, the structural representation of the compounds of the invention may include the free base form, the salt form, the zwitterionic form, or quaternary ammonium form, other of such compounds discussed above. All forms are encompassed. Thus, one aspect of the present invention is the pharma- ceutical use of the compounds of the present invention. It is provided in the form of an acceptable salt, zwitterionic complex or quaternary ammonium complex. Those skilled in the art will recognize examples of compounds of the invention capable of forming such complexes, examples of which include , the tetracoordinate nitrogen is quaternized or protonated, and the charged nitrogen form is attached by the associated anion. The term "pharmaceutical acceptable salt" refers to a compound that is capable of being stabilized. have similar or superior efficacy to the free base form of the compound and are biologically or Is not otherwise undesirable (e.g., is not toxic to the recipient) (quaternary ammonium complexes and internal salts, e.g., (including zwitterionic complexes, etc.)

[0146] The formation of pharma- ceutically useful salts from basic (or acidic) pharmaceutical compounds is described, for example, in S. Berge et al., Journal of Pharmaceutical Science s(1977) 66(1) 1-19;P.Gould,International J. of Pharmaceutics(1986) 33 201-217;An derson et al,The Practice of Medicinal C hemistry (1996), Academic Press, New York The Orange Book (Food & Drug Administration on, Washington, DC, on its website; and P. Hei nrich Stahl, Camille G. Wermuth (Eds.), Hand book of Pharmaceutical Salts:Properties, Selection,and Use,(2002)Int'l.Union of P Review and Applied Chemistry, pp.330-331 No. 6,393,636, the disclosures of which are incorporated herein by reference.

[0147] The present invention contemplates both the free base form and all available salts of the compounds of the present invention. The salts are those generally recognized as safe for use in the preparation of pharmaceutical formulations, and and the like, which are now within the scope of ordinary skill in the art, and are useful in the preparation of pharmaceutical formulations. "generally recognized as safe" for use in "safe as prescribed" and are described herein. In the present specification, the term "pharmaceutically acceptable salts" is used. As will be understood, the free base compounds may be used without modification. By controlling the isolation conditions of the compound during the synthesis or from a salt form of the compound of the present invention, It may be prepared by neutralization and ion exchange.

[0148] Examples of pharma- ceutically acceptable acid salts include, but are not limited to, trifluoroacetamide. Acetate salts include acetate, adipate, alginate, ascorbate, aspartate, Lutetate, benzoate, benzenesulfonate, bisulfate, borate, butyrate citrate, camphorate, camphorsulfonate, cyclopentanepropionate Gluconate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoside Heptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate Ethyl, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate phonate, lactate, maleate, methanesulfonate, methylsulfate, 2-naphthalene Phthalene sulfonate, nicotinate, nitrate, oxalate, pamoate, pec Tinate, persulfate, 3-phenylpropionate, phosphate, picrate , pivalate, propionate, salicylate, succinate, sulfate, sulfone tartrates (such as those mentioned herein), thiocyanates, Includes toluenesulfonate (also known as tosylate), undecanoate, and the like.

[0149] Examples of pharma- ceutically acceptable base salts include, but are not limited to, ammonium salts. Salts, alkali metal salts, e.g. sodium, lithium and potassium salts, alkaline earth metal Salts, such as calcium and magnesium salts, aluminum salts, zinc salts, organic bases (e.g. salts with organic amines, e.g. benzathine, diethylamine, dicyclohexylamine, Formed using hydrabamine (N,N-bis(dehydroabietyl)ethylenediamine) ), N-methyl-D-glucamine, N-methyl-D-glucamide, t-butylamine, Piperazine, phenylcyclohexylamine, choline, tromethamine salts, and a The basic nitrogen-containing group includes salts with amino acids, such as salts with arginine and lysine. can be converted to a monium ion, or, for example, a lower alkyl halide (e.g., methyl , ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfides esters (e.g. dimethyl, diethyl, dibutyl and diamyl sulfates), long chain halide dopants (e.g. decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides) agents such as aryl halides (e.g., benzyl and phenethyl bromides), and aralkyl halides (e.g., benzyl and phenethyl bromides). It can be quaternized using

[0150] The term "pharmacologically acceptable anion" refers to an anion that is present in a compound to form a pharma- ceutically acceptable salt. Refers to a suitable anion.

[0151] Further examples of pharma- ceutically acceptable salts that can be used in the present invention include, but are not limited to, does not include, but does include, fluoride, chloride, bromide and iodide.

[0152] In general, salts of the compounds are intended to be pharma- ceutically acceptable salts that are within the scope of the present invention. It is illustrated.

[0153] The terms "purified," "in purified form," or "in isolated and purified form" referring to a compound "In its original form" means isolated from a synthetic process or from a natural source, or a combination thereof. It refers to the physical state of the compound after purification. Hence, the term "purified" "In purified form" or "in isolated and purified form" are used herein. or after being obtained from a purification process or processes well known to those skilled in the art. or known to those skilled in the art. The term "compound" refers to the physical state of said compound in sufficient purity. The compounds of the present invention may be in any form. These include those in situ in the reaction mixture, as well as those isolated by conventional techniques. Also included are isolated and purified forms obtained by the above steps. Polymorphic forms of solvates and prodrugs are also included.

[0154] Certain compounds of the invention may exist in different tautomeric forms, for example as limited Although not limited to these, they do exist in the ketone / enol tautomeric form, the imine-enamine tautomeric form, and and for example heteroaromatic ring forms, such as the moiety: [ka]

[0155] It may be in the form:

[0156] Similarly, unless otherwise indicated, the structures of any tautomeric forms of compounds that exhibit tautomerism. The presentation of a representation is meant to encompass all such tautomeric forms of the compound. Thus, the compounds of the present invention, their salts, and their solvates and prodrugs are Where an alkyl group can exist in different tautomeric forms or in equilibrium between such forms, All such forms of the compound are encompassed by and within the scope of the present invention.

[0157] In another aspect, the present invention provides pharmaceutical compositions comprising one or more compounds of the present invention. As used herein, the term "pharmaceutical composition" refers to a pharmaceutical composition that contains at least one pharmacologic agent. A combination of specified ingredients in specified amounts, including a mixture and at least one additive. and any product resulting directly or indirectly from the combination of specified ingredients in specified amounts. The term "polysaccharide" is intended to encompass both synthetic and non-synthetic products.

[0158] As will be appreciated by those skilled in the art, an additive may itself exert an active pharmaceutical effect. To adapt the composition to a particular route of administration or to aid in processing the composition into a dosage form without Generally, the composition contains the active ingredient to be administered. Depending on the route of administration and the characteristics, it may contain two or more excipients. Examples of additives that impart properties that facilitate tableting include, but are not limited to, additives intended for tableting. Lubricants or pressing aids in powdered pharmaceuticals that are to be mixed and used when the active ingredient is in emulsion form The composition may be adapted to the desired route of administration. Examples of additives that can be used include, but are not limited to, additives that can be used to inhibit absorption from the digestive tract in the case of oral administration. It is an absorption enhancer that promotes absorption and is used for transdermal administration. For oral or mucosal administration, penetration enhancers ), e.g., as used in adhesive skin "patches" or compositions for buccal administration It is.

[0159] Regardless of the function that an additive performs in the composition, additives are collectively referred to herein as "supports." Typically, the formulation contains up to about 95 percent active ingredient and the balance carrier. A balance carrier may be included, although formulations with different ratios may also be prepared. Generally, an acceptable pharmaceutical composition is one that is tolerated for a period of time by the subject to which the composition is administered. and capable of providing therapeutic serum levels of the active agent within an acceptable temperature range. Based on the route of administration, the composition may be formulated so that it retains biological activity during an acceptable period of storage. and providing an effective amount of a PCSK9 antagonist in a separate dosage form with a volume acceptable for administration. The composition contains a suitable concentration of active agent that can

[0160] As used herein, a pharmaceutical composition refers to a bulk composition, i.e., individual components for administration. Formulated materials that have not yet been formed into dosage units and the components contained within the individual dosage units. The term "compositions" refers to both compositions that

[0161] While the compositions of the present invention may be used in bulk form, for most applications the compositions will be administered to the patient. and each dosage form contains an effective amount of said It is understood to include an amount of a selected composition that contains one or more compounds of formula I. Examples of suitable dosage forms include, but are not limited to: (i) dosage forms adapted for oral administration; Examples include liquid, gel, powder, solid or semi-solid pharmaceutical compositions filled into capsules. The drug is filled or compressed into a tablet and additionally has one or more coatings that modify its release characteristics. formulations having coatings that impart delayed release or have sustained release properties (ii) dosage forms adapted for administration through the tissues of the oral cavity, e.g., fast dissolving tablets, lozenges , a liquid, gel, sachet or needle array suitable for providing intramucosal administration; i) Formulations adapted for administration via the mucous membranes of the nose or upper respiratory cavity, e.g., nasal or intrabronchial (iv) solutions, suspensions or emulsions adapted for dispersion in the skin; (v) dosage forms adapted for intradermal administration, e.g., patches, creams or gels; (vi) dosage forms adapted for intravenous (IV) injection, e.g., IV injections (vii) dosage forms adapted for intramuscular (IM) administration, e.g. For example, injectable solutions or suspensions, which may be adapted to form depots with sustained release properties. (viii) dosage forms suitable for intravenous (IV) administration, e.g., liquids; or as a suspension, e.g., an IV solution or concentrate injected into a saline IV bag. (ix) allowing the compound to diffuse into the surrounding tissue, thereby providing sustained therapeutic serum levels. This includes long-term administration by implanting a rod or other device that provides (x) a dosage form adapted for subcutaneous administration; or (x) a dosage form adapted for delivery through the rectal or vaginal mucosa. The dosage forms include, for example, suppositories.

[0162] Pharmaceutical compositions can be solid, semi-solid or liquid. Solid, Semi-Solid and Liquid The formulations can be adapted for various modes of administration, examples of which include, but are not limited to, These include, but are not limited to, powders, dispersible granules, mini-tablets and beads, which may be, for example, tableted. In addition, it can be used for liquid form preparations and for direct administration. Examples of suitable liquids include, but are not limited to, solutions, suspensions, and emulsions. For example, but not exclusively, those intended for oral ingestion, inhalation or intravenous (IV) administration. The formulations may be administered, for example, but not limited to, via IV drip or infusion pump. Administration, intramuscularly (IM), e.g., intramuscularly as a bolus released over a sustained period of time Administration (IM), formulations intended for direct IV injection, or compatible with the subcutaneous route of administration It can be used in the preparation of formulations.

[0163] Other possible routes of administration include intranasal administration, or administration to other mucous membranes. Formulations prepared for administration to various mucous membranes also include those suitable for such administration. Additional components may be included, such as viscosity modifiers.

[0164] In some embodiments, compositions suitable for use in solid oral dosage forms, e.g., Compositions suitable for use in tablets or quickly dissolving orally soluble formulations include those comprising the compounds of the present invention. or a salt thereof is the preferred route of administration, but the compositions of the present invention may also be administered via the other routes mentioned above. For example, the composition may be formulated for administration via aerosol, e.g., via inhalation. or aerosol formulations suitable for administration via the nasal mucosa, which include solutions and powder forms. These may include solids in the form of a pharma- ceutically acceptable propellant, e.g., an inert compressed gas, For example, it may be combined with nitrogen. It may also be added to a suspension or solution immediately prior to use, for example, orally or Also included are solid form preparations that are intended to be converted to suspensions or solutions for parenteral administration. Examples of such solid forms include, but are not limited to, lyophilized formulations and and liquid formulations absorbed in a solid absorption medium.

[0165] For example, the compounds of the invention may also be formulated in the form of, for example, a liquid, suppository, cream, foam, gel or From a rapidly dissolving solid form, the composition may be deliverable transdermally or transmucosally. The composition may take the form of a cream, lotion, aerosol and / or emulsion. Any transdermal patch known in the art may be used, such as a patch for administering a pharma- ceutical active compound to a patient. or a reservoir containing a pharma- ceutical active compound in solid or liquid form. It will be appreciated that the compositions may be provided in a unit dosage form such as an incorporated patch.

[0166] Examples of pharma- ceutically acceptable carriers and methods of manufacturing the various compositions referred to above can be found in AG ennaro(ed.), Remington: The Science and Pr. Actice of Pharmacy, 20 th Edition, (2000), L ippincott Williams & Wilkins,Baltimore,M Additional examples of publications dealing with formulation issues can be found in: Pharmaceutical compositions can be formulated by a number of strategies known in the art. McGoff and Scher, 2000 Solution Formula tion of Proteins / Peptides:In-McNally,EJ .,ed.Protein Formulation and Delivery.Ne w York, NY:Marcel Dekker;pp.139-158;Akers &Defilippis,2000,Peptides and Proteins a s Parenteral Solutions.In-Pharmaceutical Formulation Development of Peptides and Proteins.Philadelphia,PA:Taylor and Fra ncis;pp.145-177;Akers et al., 2002,Pharm.Biotec See H.Nol. 14:47-127.

[0167] In another embodiment, the present invention provides a method for the treatment of PCSK9 comprising administering to a subject the method of the present invention ... The present invention provides a method of using a PCSK9-specific antagonist compound, the method comprising: The use of the term "antagonize" is used throughout this application to refer to the activation of the affected tissue ( (class I) opposes or inhibits one or more functions of PCSK9 in affected tissues refers to the supply of a substance that opposes, counteracts, neutralizes, or reduces a function. Inhibition or antagonism of one or more of the functional properties associated with PCSK9 is known in the art. Known methodologies (e.g., Barak & Webb, 1981 J. Cell Biology ol.90:595-604;Stephan&Yurachek,1993 J.Li pid Res.34:325330; and McNamara et al., 2006 Clin ica Chimica Acta 369:158-167), and This can be readily determined according to the methodology described herein. , a decrease in PCSK9 activity compared to that seen in the absence of an antagonist, or e.g. For example, compared to the activity observed in the presence of a control antagonist of irrelevant specificity. The reduction in PCSK9 activity observed in the present invention is preferably achieved by using a PCSK9-specific Antagonists can inhibit the function of PCSK9, including but not limited to, those disclosed herein. More preferably, to the extent that a measured parameter, including the activity of the For example, the measured parameter is at least 20%, 30%, 40%, 50%, 60%, 70% , to the extent that PCSK9 function is reduced by 80%, 90% and 95%. Such inhibition / antagonism may be useful in treating certain phenotypes, diseases, or disorders in which PCSK9 function is adversely affected in a subject. or contributes at least partially to the symptoms.

[0168] In one embodiment, the present invention provides a method for antagonizing the activity of PCSK9, comprising: The method can be influenced by PCSK9 (i.e., those that express the LDL receptor and The cells, cell populations or tissue samples (including and / or containing PCSK9) as disclosed herein may be subjected to A specific antagonist that binds to PCSK9 in the presence of the antagonist and inhibits P contacting under conditions that allow inhibition of CSK9 inhibition of cellular LDL uptake; In some embodiments of the invention, the method includes such a method, wherein the cell is a human cell. Additional embodiments of the invention include methods where the cell is a mouse cell. .

[0169] In one embodiment, the present invention provides a method for antagonizing the activity of PCSK9 in a subject. The method comprises administering to a subject a therapeutically effective amount of a PCSK9-specific antagonist of the invention. In some embodiments, to antagonize PCSK9 function, The method includes administering to a subject a disease, disorder or condition associated with PCSK9, as defined herein. or for the treatment of, or for the treatment of, any of the following conditions that could benefit from the effects of a PCSK9 antagonist: The present invention is intended to provide treatment for a disease, disorder or condition that may be caused by a complication of the present invention.

[0170] The present invention therefore provides a method for the treatment of various conditions in which it is desirable to antagonize PCSK9 function. The present disclosure contemplates the use of the PCSK9-specific antagonists described herein in As used herein, the term "treatment method" refers to a method for treating a disease by causing an alteration in at least one symptom of the disease. With respect to a course of action that results in a condition, this can be preventative or therapeutic in nature. In some embodiments, the present invention provides methods for treating PCSK9-related and / or -causing disorders. for conditions that are contraindicated for PCSK9 function or for conditions that contraindicate PCSK9 function in a particular subject The present invention relates to a method of treatment for a PCSK9 disorder, the method comprising administering to a subject a therapeutically effective amount of a PCSK9 antagonist of formula I. The method includes administering a steroid compound or a pharma- ceutical acceptable salt thereof to a subject. In this condition, symptoms include atherosclerosis, hypercholesterolemia, coronary heart disease, Metabolic syndrome, acute coronary syndrome, or related cardiovascular and cardiometabolic diseases It may be a condition, or it may be a disease state or condition in which PCSK9 activity is contraindicated. .

[0171] The method of treatment according to the present invention comprises administering to an individual a therapeutically (or prophylactically) effective amount of a P The term "therapeutically effective" refers to the administration of a CSK9 specific antagonist. The use of "prophylactically effective" or "prophylactically effective" refers to a compound that, at the intended dose, provides the desired therapeutic and / or It refers to the amount necessary to achieve a prophylactic effect for a desired period of time. The present invention may provide a method for treating a condition, comprising the steps of: alleviating, ameliorating, reducing, or ceasing at least one symptom associated with the condition being treated; As one of skill in the art will appreciate, these amounts may vary depending on, but are not limited to, the individual's disease state, PCSK9-specific antigens that elicit the desired effects in individuals of different ages, sexes, and weights Responses vary depending on a variety of factors, including the potency of the antagonist. The efficacy and safety of the compound may be documented by in vivo non-human animal studies and / or further supported by clinical trials. It can be held.

[0172] In some embodiments, the PCSK9 antagonist compounds of the invention are It is preferred to administer the compound in the form of a pharmaceutical composition as described herein.

[0173] Dosing of antagonist therapeutic agents is well within the capabilities of one of ordinary skill in the art (see, for example, Le derman et al., 1991 Int. J. Cancer 47:659-664; Shawe et al., 1991 Antibody, immunoconjugates and d Radiopharmaceuticals 4:915-922) The severity of the disease will depend on a number of factors, including but not limited to the condition of the patient, the area being treated, administration, Based on the factors mentioned above, including the route and the desired treatment, e.g., prophylactic or acute treatment. A physician or veterinarian of ordinary skill will determine a therapeutically effective amount of the antagonist. It can be easily determined and prescribed.

[0174] The subject is a person in need of or desiring treatment for an existing disease or medical condition. As used herein, a "need" for treatment of an existing condition. The subject will be informed of both the determination of need by the health care provider and the subject's desire for such treatment. When the compound or salt thereof is provided in combination with one or more other active agents, "Administer" and variations thereof refer to the contemporaneous or simultaneous administration of a compound or a salt thereof and another agent, respectively. It is understood to include administration in a single dose, or in a series of separate doses over a period of time. When the agents of the combination are administered simultaneously, they are administered together in a single composition. or may be administered separately. A "combination" of active agents refers to all of the active agents The composition may be a single composition containing one or more active agents, or multiple compositions each containing one or more active agents. In the case of two active agents, the combination can be a single formulation containing both agents. The composition may be one of the above, or two separate compositions each containing one agent; In the case of active agents, the combination may be a single composition containing all three agents, a three-component composition each containing one agent, or one separate composition, or two compositions, one of which contains two agents and the other of which contains three agents. A second agent may be included, etc.

[0175] The compositions and combinations of the present invention are preferably administered in an effective amount. antagonizing PCSK9 and thereby eliciting the desired response (i.e., in animals or In humans, the effects include, but are not limited to, atherosclerosis, hypercholesterolemia, , coronary heart disease, metabolic syndrome, acute coronary syndrome, and related cardiovascular Treatment of conditions related to or affected by PCSK9 function, including diseases and cardiometabolic conditions By "therapeutic effector agent" is meant an amount of active compound sufficient to induce a therapeutic response in a particular site or administration.

[0176] The actual dosage employed will vary depending on the requirements of the patient and the severity of the condition being treated. Determination of the proper dosage regimen for a particular situation can be found, for example, in the standard literature. For example, the “Physicians' Desk Reference” (PDR ), for example, the 1996 edition (Medical Economics Com Pany, Montvale, NJ 07645-1742, USA), Physici an's Desk Reference, 56 th Edition, 2002 (Me dical Economics company, Inc. Montvale, NJ 07645-1742) or Physician's Desk Refe rence,57 th Edition, 2003 (Thompson PDR, Mon The present invention relates to a method for the preparation of a medicament for use in a pharmaceutical composition, as described in the publication by Tvale, NJ 07645-1742. these disclosures are incorporated herein by reference thereto. For convenience, the total daily dosage is divided and administered in portions throughout the day as needed. The delivery may be continuous or continuous.

[0177] PCSK9-specific antagonists can be administered orally or by injection, but are not limited to these. Administered by intravenous injection, subcutaneous injection, intraperitoneal injection or intramuscular injection (specific embodiments of which include intravenous injection, subcutaneous injection, intraperitoneal injection or intramuscular injection) or administration by inhalation, intranasal administration, or topical administration. By any route of administration understood in the art, alone or as an adjunct in treating an individual. PCSK9-specific antagonists may be administered to individuals in combination with other agents designed to inhibit PCSK9. The agonists are also available in injection devices, injection pens, needle-free devices; and subcutaneous patch delivery systems. The route of administration may be, but is not limited to, the desired physiological The determination should be based on many considerations, including chemical properties, as will be understood by those of skill in the art. be.

[0178] One or more additional pharmacologically active agents may be administered in combination with the compounds of formula I. The active agent (or active agents) is a pharma- ceutical active agent (or agents) that is active in the body and is different from the compound of formula I ( or active agents), which are converted into a pharmacologic active form after administration. and prodrugs thereof, as well as free acids, free bases and pharma- ceutical acceptable salts of said additional active agents. In general, the term "antihypertensive agent", "antiatherosclerotic agent", "antihypertensive ... Any suitable antidiabetic and / or antiobesity agent, such as a lipid modifying compound, Any suitable additional active agent or active agents may be used in combination with a compound of Formula I in a single dosage formulation. They may be used in combination (fixed dose drug combinations) or in combination with simultaneous or sequential administration of the active agents. The active agents may be administered to a subject in one or more separate dosage formulations allowing for co-administration of separate active agents. ).

[0179] Examples of additional active agents that may be used include, but are not limited to, angiotensin II Enzyme-converting enzyme inhibitors (e.g., alacepril, benazepril, captopril, cerenapril, , Cilazapril, Delapril, Enalapril, Enalaprilat, Hosinopril, Imipril Dapril, lisinopril, moveltipril, perindopril, quinapril, ramipril, spirapril, temocapril or trandolapril), angiotensin II receptor antagonists antagonists (e.g., losartan, i.e., COZAAR®, valsartan Candesartan, olmesartan, telmesartan, and any of these agents used in combination with hydrochlorothiazide, e.g., HYZ AAR®, etc.); neutral endopeptidase inhibitors (e.g., thiorphan and and phosphoramidon), aldosterone antagonists, aldosterone synthase inhibitors , renin inhibitors (e.g., urea derivatives of dipeptides and tripeptides (U.S. Pat. No. 5, 116,835), amino acids and derivatives (see U.S. Pat. No. 5,095,1 19 and 5,104,869), amino acid chains linked by non-peptide bonds (U.S. Patent No. 5,114,937), dipeptide and tripeptide derivatives, peptidyl peptidyl β-aminoacyl aminodiol carbamates, and small molecule renin inhibitors (including diol sulfonamides and sulfinyls), N-morpholino derivatives, N-heterocyclic alcohols and pyrrole imidazolones; Fluoro- and chloro-peptides of statin derivatives and statone-containing peptides. RORO derivative, enalkrein, RO 42-5892, A 65 317, CP 80794, ES 1005, ES 8891, SQ 34017, Ali Skillene (2(S),4(S),5(S),7(S)-N-(2-carbamoyl-2-methyl) 5-amino-4-hydroxy-2,7-diisopropyl-8-[4-methylpropyl] Toxic-3-(3-methoxypropoxy)-phenyl]-octanamide hemifumarate ) SPP600, SPP630 and SPP635), endothelin receptor antagonists phosphodiesterase-5 inhibitors (e.g. sildenafil, tadalfil (ta dalfil) and vardenafil), vasodilators, calcium channel blockers (e.g. Amlodipine, nifedipine, verapamil, diltiazem, gallopamil, nildipine nimodipine, nicardipine), potassium channel activators (e.g., nicorandil , pinacidil, cromakalim, minoxidil, aprilkalim , loprazolam), diuretics (e.g., hydrochlorothiazide), sympatholytics (sy mpatholitics, β-adrenergic blockers (e.g., propranolol, atenolol, bisoprolol, carvedilol, metoprolol or metoprolol tartate), alpha adrenergic blockers (e.g., doxorubicin, alpha-adrenergic agonists (e.g., prazosin, alpha-methyldopa, or alpha-amyclohexidine) Peripheral vasodilators (e.g., hydralazine); lipid lowering agents, e.g., HMG-Co A reductase inhibitors, such as ZOCOR® and and MEVACOR®, which act as inhibitors after administration. Nvastatin and lovastatin, and dihydroxy open acid HMG-CoA reductase Pharmaceutically acceptable salts of enzyme inhibitors, such as atorvastatin (especially LIPITOR) (calcium salt sold under the trademark CRESTO pravastatin (particularly the calcium salt sold under the trademark PRAVA CHOL®), fluvastatin (especially LE SCOL® (sodium salt), crivastatin (crivas tatin and pitavastatin; cholesterol absorption inhibitors, such as ezetimibe (Z ETIA®), and ezetimibe and any other lipid lowering agent, such as the HM Combination with G-CoA reductase inhibitors, especially simvastatin (VYTORIN®) or in combination with atorvastatin calcium; Immediate or controlled release forms and / or HMG-CoA reductase inhibitors Niacin in combination with an anti-inflammatory agent; niacin receptor agonists, e.g. acipimox and and Acifran, as well as niacin receptor partial agonists; metabolic modifiers olic altering agent, insulin and insulin mimetics Tics (e.g., insulin degludec, insulin glargine, insulin lispro), dipeptidyl peptidase-IV (DPP-4) inhibitors (e.g., sitag liptin, alogliptin, omarigliptin, linagliptin, vildagliptin) Insulin sensitizers, including: (i) PPARγ agonists stomatitis, such as glitazones (e.g. pioglitazone, AMG 131, MBX2044, Mitoglitazone, lobeglitazone, IDR-105, rosiglitazone and balaglitazone (1) PPARα / γ dual agonist Examples include ZYH2, ZYH1, GFT505, and chiglitazar. zar), muraglitazar, aleglitazar, soderglitazar and naveglitazar) (2) PPARα agonists, such as fenofibric acid derivatives (e.g., gemcitabine, Brozil, clofibrate, ciprofibrate, fenofibrate, bezafibrate (3) selective PPARγ modulators (SPPARγMs) (e.g., WO0 2 / 060388, WO02 / 08188, WO2004 / 019869, WO2004 / 020409, WO2004 / 020408 and WO2004 / 066963 (ii) PPARγ partial agonists; and (iii) biguanidine. amides, such as metformin and pharma- ceutically acceptable salts thereof, especially metformin; The hydrochloride salt, as well as sustained release formulations thereof, such as Glumetza™, Fortam et™ and GlucophageXR™; and (iii) protein Protein tyrosine phosphatase-1B (PTP-1B) inhibitors (e.g., ISIS-1137 15 and TTP814); insulin or insulin analogues (e.g., insulin Insulin detemir, insulin glulisine, insulin degludec, insulin glu Lupus, insulin lispro and inhalable formulations of each; leptin and leptin amylin and amylin analogues (e.g., pramylin Sulfonylurea and non-sulfonylurea insulin secretagogues (e.g. , tolbutamide, glyburide, glipizide, glimepiride, mitiglinide, meglitinide , nateglinide, and repaglinide); α-glucosidase inhibitors (e.g., acarbose, glucagon receptor antagonists (e.g., , MK-3577, MK-0893, LY-2409021 and KT6-971); Incretin mimetics, such as GLP-1, GLP-1 analogs, derivatives and mimetics and GLP-1 receptor agonists (e.g., dulaglutide, semaglutide, albiglutide, exenatide, liraglutide, lixisenatide, taspoglutide, CJC-1131 and BIM-51077, their intranasal, transdermal and once-weekly formulations bile acid sequestrants (e.g., colestilan, colestimide, colesebalam (colesevalam) hydrochloride, colestipol, cholestyramine, and cross-linked dextran Dialkylaminoalkyl derivatives of stran, acyl CoA: cholesterol acylate transferase inhibitors (e.g., avasimibe); anti-obesity compounds; in inflammatory conditions The agent for which it is intended to be used, e.g., aspirin, nonsteroidal anti-inflammatory drugs or NSAIDs, Glucocorticoids and selective cyclooxygenase-2 or COX-2 inhibition glucokinase activators (GKA) (e.g., AZD6370); 11β-hydro Inhibitors of hydroxysteroid dehydrogenase type 1 (see, e.g., U.S. Patent No. 6,730,693, 0, and LY-2523199); CETP inhibitors (e.g., anacetrapib, torcetrapib and evacetrapib); fructose 1,6- Inhibitors of bisphosphatases (see, e.g., U.S. Pat. No. 6,054,587; No. 6,284,748; No. 6,399,782; and No. 6,489 ,476); acetyl-CoA carboxylase-1 or 2 Inhibitors of (ACC1 or ACC2); AMP-activated protein kinase (AMPK) Activators; other G protein-coupled receptor agonists: (i) GPR-109, (ii) GPR-119 (e.g., MBX2982 and PSN821), and (iii) GPR-40 (e.g., TAK875); SSTR3 antagonists (e.g., WO 2009 / 001836); Neuromedin U receptor agonists ( By way of example, and not limitation, those disclosed in WO2009 / 042053. contains neuromedin S (NMS); SCD modulator; GPR-105 antagonists (e.g., those disclosed in WO2009 / 000087); S GLT inhibitors (e.g., ASP1941, SGLT-3, empagliflozin, dapag Liflozin, Canagliflozin, BI-10773, Ertugliflozin, Remogliflozin Remogloflozin, TS-071, Tofogliflozin, Ipraglif Rosin and LX-4211); Acyl Coenzyme A: Diacylglycerol Acyl Inhibitors of transferases 1 and 2 (DGAT-1 and DGAT-2); fatty acid synthesis Inhibitors of enzymes; acyl-coenzyme A: monoacylglycerol acyltransferase Inhibitors of MGAT-1 and MGAT-2; agonists of the TGR5 receptor (also known as GPBAR1, BG37, GPCR19, GPR131 and M-BAR) ileal bile acid transporter inhibitors; PACAP, PACAP mimetics and and PACAP receptor 3 agonists; PPAR agonists; protein tyrosine phosphatase IL-1b antibodies (e.g., XOMA052 and and canakinumab; and bromocriptine mesylate and its immediate release formulations; and other drugs useful for the treatment of the above conditions or disorders, where chemically feasible This includes the free acid, free base and pharma- ceutically acceptable salt forms of the above active agents. nothing.

[0180] The compounds of the present invention can be readily prepared according to the following reaction schemes and examples or modifications thereof. They can be readily prepared using available starting materials, reagents and conventional synthetic procedures. In these reactions, known variants can also be used. Purification of compounds using HPLC (either HPLC or MPLC, as described below) Another method for preparing the compounds of the present invention is as follows: In light of the reaction schemes and examples, it will be readily apparent to one skilled in the art. The term may be used in the illustrative schemes and / or examples herein.

[0181] ACN is acetonitrile. AcOH is acetic acid. AcO - NH4 is ammonium acetate. Boc2O is di-tert-butyl dicarbonate. Bn is benzyl. BnBr is benzyl bromide. BzCl is benzoyl chloride. CBr4 is perbromomethane. Cbz-Cl is benzyl chloroformate. DBU is 1,8-diazabicyclo[5.4.0]undec-7-ene. DCC is dicyclohexylcarbodiimide. DCE is 1,2-dichloroethane. DCM is dichloromethane. DEA is N,N-diethylamine. DIAD is (E)-diisopropyldiazene-1,2-dicarboxylate. DIEA or DIPEA is N,N-diisopropylethylamine. DMAP is 4-dimethylaminopyridine.

[0182] DMF is N,N-dimethylformamide. DMSO is dimethyl sulfoxide. EA or EtOAc is ethyl acetate. EtOH is ethanol. Et2O is diethyl ether. Fmoc is a fluorenylmethyloxycarbonyl protecting group. Fmoc-Cl is (9H-fluoren-9-yl)methyl carbonochloridate . Fmoc-D-Dap(Boc)-OH is N-alpha-(9-fluorenylmethyl) (oxycarbonyl)-N-beta-t-butyloxycarbonyl-D-2,3-diaminopropane Propionic acid. Fmoc-Osu is Fmoc N-hydroxysuccinimide ester. HATU is 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo [4,5-b]pyridinium 3-oxide hexafluorophosphate. HPLC is high performance liquid chromatography. IPA is isopropyl alcohol. LiOH is lithium hydroxide. LC / MS is liquid chromatography-mass spectrometry. Me3N is trimethylamine. MeOH is methanol. MPLC is medium pressure liquid chromatography. MsCl is methanesulfonyl chloride. NaBH(OAc)3 is sodium triacetoxyborohydride. NMR is nuclear magnetic resonance. NsCl is 4-nitrobenzene-1-sulfonyl chloride. PE is petroleum ether. Pd2(dba)3(HCCl3) is tris(dibenzylideneacetone)dipalladium (0)-Chloroform adduct. PPh3 is triphenylphosphine. PdCl2(dppf) or Pd(ii)(dppf)Cl2 is a dichloro[1,1' -bis(diphenylphosphino)ferrocene]palladium(II). Pd(dppf)Cl2CH2Cl2 is dichloro[1,1'-bis(diphenylphosphine) [phenylino)ferrocene]palladium(II) dichloromethane adduct. Pd(PPh3)4 is tetrakis(triphenylphosphine)palladium. PPT s is pyridinium p-toluenesulfonate. [Rh(OAc)2]2 is the rhodium(II) acetate dimer. RT or rt or rt is room temperature. tBuOAc is tert-butyl acetate. TEA is triethylamine. TFA is trifluoroacetic acid. TFE is tetrafluoroethylene. THF is tetrahydrofuran. Tf2O is trifluoromethanesulfonic anhydride. Teoc-OSu is 2,5-dioxopyrrolidin-1-yl (2-(trimethylsilyl) ) ethyl) carbonate. TBAF is tetrabutylammonium fluoride. TMS is tetramethylsilane. Zhan catalyst 1B is dichloro(1,3-bis(2,4,6-trimethylphenyl)-2 -imidazolidinylidene)((5-((dimethylamino)sulfonyl)-2-(1-methyl Ruthenium(II) is a 1,3-bis(ethoxy-O)phenyl)methylene-C 2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene[2 -(i-Propoxy)-5-(N,N-dimethylaminosulfonyl)phenyl[methylene Also described as ruthenium(II) dichloride]. EXAMPLES

[0183] Example 1 Preparation of Ex-01 and Ex-51 [ka] TIFF2025041712000103.tif90134

[0184] The salt forms of compounds Ex-01 and Ex-51 can be prepared as intermediates 76 and 77 according to the following scheme: and 86 (prepared below): [ka] TIFF2025041712000105.tif230168TIFF2025041712000106.tif79168

[0185] Step A: Preparation of intermediate 89 76 (1.56 g, 1.917 mmol, prepared below) in DMF (40 ml) A solution of 86 (1.506 g, 1.936 mmol, prepared below) was added to HATU (0.80 2g, 2.108mmol) and DIEA (0.670ml, 3.83mmol) were added. The resulting solution was stirred at room temperature for 50 min and then diluted with EtOAc (300 mL) and brine. The organic layer was washed with brine (2×100 mL) and partitioned between ethyl acetate and nitrogen (100 mL). The residue was purified by drying over a2SO4, concentrating, and eluting with MeOH / DCM on a silica gel column. Purification using hexane as a solvent gave 89. LC / MS: (M+1) + :1573. 8.

[0186] Step B: Preparation of Intermediate 90 A solution of 89 (0.68 g, 0.432 mmHg) in DCM (500 ml) and acetic acid (40 mL) was added. A solution of 0.222 g of Zhan catalyst-1b (0.222 g) was bubbled with N2 for 20 min. The resulting mixture was then bubbled with N2 for 20 min. The mixture was then cooled to room temperature and filtered through Celite. The filtrate was concentrated, and the residue was purified by silica gel column chromatography using MeOH / DCM as elution solvent. Purification by HPLC gave 90 (cis / trans mixture). LC / MS: (M+1) + :1545.7.

[0187] Step C: Preparation of Intermediate 91 90 (cis / trans mixture) (65 mg, 0.042 To a solution of 1.2 mmol, piperidine (0.042 ml, 0.420 mmol) was added. The resulting solution was stirred at room temperature for 1 h, then concentrated, and the residue was dissolved in acetonitrile (4 mL). The residue was dried under high vacuum for an additional 30 min to give 91(cis). and trans mixture). LC / MS: (M+1) + :1324.0.

[0188] Step D: Preparation of Intermediate 92 91 (cis / trans mixture) (548 mg, 0.41 4 mmol) and 88 (227 mg, 0.455 mmol, prepared below) were added to a solution of H ATU (181 mg, 0.476 mmol) and DIEA (0.166 ml, 0.95 The resulting solution was stirred at 0° C. for 1 hour, and the solution was then reverse-coupled on a C18 column. The mixture was dissolved in acetonitrile (0.05% TFA) / water (0.05% TFA) by phase MPLC. Purification using 1,2-dichlorophenyl ether as the eluent gave 92 (cis / trans mixture). S: (M+1) + :1803.5.

[0189] Step E: Preparation of Intermediate 93 92 (700 ml) in THF (20 ml), MeOH (6 ml) and water (6 ml) at 0 °C A solution of 1N LiOH (3.11 ml, 3.11 The solution was stirred at 0° C. for 23 h. The pH was neutralized to 7-8 by addition of Cl, the volatile components were evaporated, and the aqueous layer was acidified to pH 5. The mixture was purified by reversed-phase MPLC on a C18 column using acetonitrile (0.05% T 93 was purified using 0.05% TFA / water as the elution solvent. The salt was prepared by dissolving 93 T in water (70 mL) and acetonitrile (70 ml) at 0 °C. A solution of FA salt (427 mg, 0.257 mmol) was added to 0.1 N HCl (13.5 ml (1.350 mmol) was added dropwise and the resulting solution was stirred at 0° C. for 5 min and then frozen. Freezing and drying gave 93 as the HCl salt. LC / MS: (M+1) + :1567 .1.

[0190] Step F: Preparation of Intermediate 94 of 93 (200 mg, 0.125 mmol) as the HCl salt in DMF (30 mL) To the solution was added HATU (56.9 mg, 0.150 mmol). The resulting solution was cooled to room temperature. The mixture was stirred at rt for 30 min and then diluted with DCM (400 mL) followed by DIEA (0.065 The resulting solution was stirred at room temperature for 1 hour and the volatile components were removed. The mixture was evaporated on a rotary evaporator and the resulting DMF solution was purified by reversed-phase MPLC. Acetonitrile (0.05% TFA) / water (0.05% TFA) was used as the elution solvent. Purification gave 94. LC / MS: (M+1) + :1549.2.

[0191] Step G: Preparation of Intermediate 95 A solution of 94 (14 mg, 9.04 μmol) in MeOH (20 ml) was diluted with 10% Pd / C (1.924 mg, 1.808 μmol) was added, and the resulting mixture was heated under a H balloon The mixture was filtered through Celite, and the filtrate was concentrated. Condensation gave intermediate compound 95. LC / MS: (M+1) + :1550.9.

[0192] Step H: Preparation of Ex-01 The intermediate compound 95 (26 mg, 0.017 mmol) prepared in the previous step was , was dissolved in DCM (2 ml). To this solution was added TFA (6 mL, 78 mmol); The resulting solution was stirred at room temperature for 30 min, then concentrated and the residue was dissolved in DCM (3 mL). Then, it was treated with 4N HCl in dioxane (0.042 mL, 0.168 mmol) and resuspended in 100 mL of 100 mL of HCl. The crude product Ex-01 was purified by reversed-phase H PLC was performed using acetonitrile (0.1% formic acid) / water (0.1% formic acid) as the elution solvent. The formate form of Ex-01 was obtained by purification using LC / MS: (M+1). + :1394.4.

[0193] Step I: Preparation of Ex-51 To a solution of intermediate compound 94 (30 mg, 0.019 mmol) in DCM (2 ml), TFA (4 ml, 51.9 mmol) was added and the reaction mixture was stirred at ambient temperature for 30 minutes. It was then concentrated. The residue was dissolved in DCM (2 mL) and HCl (4N in dioxane) (0 0.048 ml, 0.194 mmol) and concentrated to give Ex-51 as the HCl salt. The compound was purified by reverse phase HPLC in acetonitrile (0.1% formic acid). The formate form of Ex-51 was obtained by purification using water (0.1% formic acid) as the mobile phase. Provided. LC / MS: (M+1) + :1392.0.

[0194] The following scheme and procedures were used to synthesize intermediates 76, 86 and 87 used in the above procedures. and 88 were prepared.

[0195] Preparation of intermediate 68 used in the preparation of intermediate 76 [ka]

[0196] Step A: Preparation of intermediate compound 65 (2S,3S)-3-hydroxypyrrolidine-2 in dioxane (100 ml) at 0° C. -A suspension of carboxylic acid (5.32 g, 40.6 mmol) was added to sodium hydroxide (122 mM l, 122 mmol) was added, followed by benzyl chloroformate (6.50 ml, 44. 6 mmol) was added dropwise. The resulting suspension was stirred at 0°C for 5 h. The volatile components were After removal, the aqueous layer was acidified to pH 3 and then washed with 30% IPA / DCM (200 mL). The aqueous layer was partitioned between brine (50 mL) and further diluted with 30% IPA / DCM (2 × 10 The combined organic layers were dried over Na2SO4 and concentrated to give (2 S,3S)-1-((benzyloxy)carbonyl)-3-hydroxypyrrolidine-2- To give the carboxylic acid (65). LC / MS: (M+1) + :266.1.

[0197] Step B: Preparation of intermediate compound 66 To a solution of 65 (7.48 g, 28.2 mmol) in MeOH (80 ml), Diazomethane (70.5 ml, 141 mmol) was added dropwise and the resulting solution was heated at room temperature. The solution was stirred for 10 minutes and then quenched by the dropwise addition of acetic acid (approximately 400 uL). Concentrate and purify the residue on a silica gel column using EtOAc / hexane as eluent. This gave 66. LC / MS: (M+1) + :280.1.

[0198] Step C: Preparation of intermediate compound 67 A solution of 66 (4.81 g, 17.22 mmol) in DCM (200 mL) was agitated for 3 h with N2. The mixture was then bubbled for 10 minutes, followed by addition of rhodium(ii) acetate dimer (0.761 g, 1. The mixture was cooled in an ice-water bath and tert-butyl diazoacetate was added. C. to 100.degree. C. The resulting mixture was stirred at 0.degree. The reaction was quenched by the addition of water (100 mL) and the mixture was stirred at RT for 1.5 h. The extract was extracted with DCM (3×100 mL) and the combined organic layers were dried over Na2SO4 and concentrated. The residue was eluted in acetonitrile (0.05% TFA) / water (0.05% TFA) as the elution solvent. The product was purified by reversed-phase MPLC using 1,2-dichloro-2,4-diphenyl-2,4-tetrahydrofuran (A). The product-containing fractions were concentrated and The aqueous layer was extracted with DCM (2×100 mL). The combined organic layers were dried over Na2SO4. and concentrated to give 67. LC / MS: (M+1) + :394.2.

[0199] Step D: Preparation of intermediate compound 68 A solution of 67 (3.72 g, 9.46 mmol) in MeOH (80 ml) was diluted with 10% P d / C (0.805 g, 0.756 mmol) was added, and the resulting mixture was heated under a H balloon The mixture was subjected to hydrogenation at ambient temperature for 2 hours and then filtered through Celite. Concentration gave 68. LC / MS: (M+1) + :259.9.

[0200] Preparation of intermediate compound 76 [ka] TIFF2025041712000109.tif158165

[0201] Step A: Preparation of intermediate 69 (in THF (20 ml), MeOH (10 mL) and water (20.00 ml) at 0 °C S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3 -(5-Fluoro-1H-indol-3-yl)propanoic acid (3 g, 6.75 mmol) ) was added NaOH (20.25 ml, 20.25 mmol) and the resulting solution was The mixture was stirred at ambient temperature for 4 hours and then the volatile components were evaporated. 0 mL) and water (20 mL) were added, and the resulting solution was cooled to 0° C. and BocO(1 The resulting solution was stirred at 0° C. for 3 hours. The volatile components were removed and the aqueous layer was extracted with Et2O (3 x 40 mL) and acidified to pH 3. and then washed with DCM (3 × 100 mL), followed by 30% IPA / DCM (2 × 80 mL). The combined organic layers were dried over Na2SO4 and concentrated to give 69. LC / MS: (M+1) + :322.9.

[0202] Step B: Preparation of intermediate 70 To a solution of 69 (2.079 g, 6.45 mmol) in DMF (40 mL) at 0 °C, 60% NaH in xanthan (0.568 g, 14.19 mmol) was added, and the resulting solution was stirred at 0° C. for 50 min, followed by addition of allyl bromide (1.172 mL, 13.54 mmol). The resulting solution was stirred at 0° C. for 1.5 hours and then diluted with 1N HCl (ca. 3. The solution was then quenched by the addition of EtOAc (200 mL) and water ( 100 mL), and the organic layer was washed with brine (2×100 mL) and The residue was purified by silica gel column chromatography using MeOH / DCM as eluent. Purification using ethyl acetate gave 70. LC / MS: (M+1) + :363.0.

[0203] Step C: Preparation of Intermediate 71 70 (2.239 g, 6.18 mmol) and 68 (1.8 A solution of HATU (2.82 g, 7.41 mmol) and and DIEA (2.59 ml, 14.83 mmol) were added, and the resulting solution was stirred at ambient temperature for 1 The mixture was partitioned between EtOAc (200 mL) and brine (100 mL). The organic layer was washed with brine (3×100 mL), dried over Na2SO4, and concentrated. The residue was purified on a silica gel column using EtOAc / hexane as the eluent. This gave 71. LC / MS: (M+1) + :604.2.

[0204] Step D: Preparation of Intermediate 72 71 (2.83 mL) in CH2Cl2 (20 mL) and tBuOAc (30 mL) at 0 °C g, 4.69 mmol) was added to a solution of methanesulfonic acid (1.218 ml, 18.75 mm ol) was added and the resulting solution was stirred at 0° C. for 16.5 h and then at ambient temperature for 2.5 h. The solution (72) was used directly in the next step. LC / MS: (M+1) + :5 04.2.

[0205] Step E: Preparation of Intermediate 73 (S)-2-((((9H-fluoren-9-yl)methoxy) )carbonyl)amino)-3-(3-(((tert-butoxycarbonyl)amino)methyl A solution of 1,2-diphenyl-2,4-trimethylphenylpropanoic acid (2.66 g, 5.16 mmol) was added to HATU ( .961g, 5.16mmol) and DIEA (5.32ml, 30.5mmol) The resulting solution was stirred at room temperature for 30 minutes, and then cooled in an ice bath with the solution of 72 prepared above. The resulting solution was stirred at ambient temperature for 1 h. The volatile components were removed by rotary evaporation. The residue was eluted with acetonitrile (0.05% TFA) / water as the elution solvent. The mixture was purified by reversed-phase MPLC using 0.05% TFA. Concentration on a tumble evaporator gave 73. LC / MS: (M+1) + :1002.1.

[0206] Step F: Preparation of Intermediate 74 A solution of 73 (3.235 g, 3.23 mmol) in DCM (4 ml) was diluted with TFA ( 0.46 mL, 97 mmol) was added and the resulting solution was stirred at ambient temperature for 1 hour, then concentrated The residue was dissolved in DCM (10 mL) and diluted with 4N HCl (3.23 mL) in dioxane. ml, 12.91 mmol), then concentrated and the residue was diluted with acetonitrile (100 ml 74. LC / MS: (M+1) + :846.1.

[0207] Step G: Preparation of Intermediate 75 To a solution of 74 (2.85 g, 3.23 mmol) in DMF (45 ml), HATU( 1.474 g, 3.88 mmol) was added and the resulting solution was stirred at ambient temperature for 30 minutes. It was then diluted with DCM (600 ml), followed by DIEA (1.692 ml, 9.69 mm The resulting solution was stirred at ambient temperature for 1 hour. The solution was concentrated to give a residue. The residue was eluted with acetonitrile (0.05% TFA) / water (0.05% TFA) as the elution solvent. The product was purified by reversed-phase MPLC on a C18 column using Concentrate and partition the aqueous layer between DCM (200 mL) and saturated NaHCO3 (200 mL). The aqueous layer was extracted with DCM (2×100 mL) and the combined organic layers were washed with Na2SO4 Drying and concentration gave 75. LC / MS: (M+1) + :828.1.

[0208] Step H: Preparation of intermediate compound 76 75(1:1) in THF (60 ml), MeOH (30 ml) and water (20 ml) at 0 °C A solution of 1.93 g, 2.331 mmol) was added to a 1N LiOH solution (9.9 ml, 9.9 0 mmol) was added dropwise and the resulting solution was stirred at 0° C. for 16 h, then HCl (1 The reaction was quenched by the addition of N (9.9 mL). The volatile components were removed by rotary evaporation. The mixture was evaporated and the solution was added with acetone (60 ml), sodium carbonate (0.371 g , 3.50 mmol) and Fmoc-Osu (0.802 g, 2.378 mmol). The resulting solution was stirred at 0° C. for 6 h, and the volatile components were removed on a rotary evaporator. The aqueous layer was then acidified to pH 4 and then diluted with 30% IPA / DCM (3 × 100 mL The combined organic layers were dried over Na2SO4, concentrated, and the residue was purified by silica gel column chromatography. Purification on a column using MeOH / DCM as the elution solvent gave 76. LC / MS:(M+1) + :814.2.

[0209] Alternative preparation of intermediate compound 75b and intermediate compound 76B therefrom: [ka] TIFF2025041712000111.tif54170

[0210] Step B is eliminated and the final step G is to replace the Fmoc protecting group with a Boc protecting group. The general procedure described above for the preparation of intermediate compound 75 can generally be followed, except that Thus providing intermediate compound 75a. The procedure for 75a and 76B is , as described below.

[0211] Step C: Preparation of intermediate compound 71a A solution of 69 (5.00 g, 15.5 mmol) in DMF (40.0 mL) at -50 °C In 68, HATU (5.90 g, 15.5 mmol) and DIEA (4.01 g, 3 1.0 mmol) was added and the reaction mixture was stirred at -50°C for 3 h. The final solution was diluted with water (5 The mixture was quenched with 1 mL of ethyl acetate, concentrated under reduced pressure, and the residue was purified by reverse phase column chromatography on C18. (eluted with a gradient of acetonitrile / water + 0.01% ammonium bicarbonate) Purification by LCMS (ESI) gave 71a. 28 H 38 FN3O8[ M+H]+ calculated: 564.3, found 564.2.

[0212] Step D: Preparation of intermediate compound 72a A solution of 2N HCl in dioxane (100 mL) and THF (100 mL) at room temperature To the mixture was added 71a (8.40 g, 14.9 mmol) and the reaction mixture was stirred for 5 h. The final solution was concentrated under reduced pressure to give 72a. LCMS (ESI) C 23 H3 Calculated for 1ClFN3O6[M-HCl+H]+: 464.2, found 464.3.

[0213] Step E: Preparation of intermediate compound 73a Dissolution of 72a (800 mg, 1.60 mmol) in DMF (10.0 mL) at -50 °C The solution was added with (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amine. (((tert-butoxycarbonyl)amino)methyl)phenyl)propane propanoic acid (827 mg, 1.60 mmol), HATU (608 mg, 1.60 mmol) ) and DIEA (620 mg, 4.80 mmol) were added, and the mixture was incubated at -50 °C for 3 h. The resulting solution was diluted with water (50 mL) and the aqueous layer was washed with EtOAc (3×100 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (1 in PE). Purification by hexanes (eluted with a gradient of 0.05%-60% EtOAc) gave 73a. LCMS(ESI) C 53 H 60 FN5O 11 Calculated [M+H]+: 962.4; Measured value: 962.6.

[0214] Step F: Preparation of intermediate compound 74a To a solution of 73a (3.00 g, 3.12 mmol) in DCM (15.0 mL) at room temperature Then, TFA (15.0 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. Concentration under reduced pressure and co-evaporation with toluene and DCM gave 74a. S(ESI)C 46 H 45 F4N5O 11 Calculated value of [M-TFA+H]+: 806.3 , measured value 806.7.

[0215] Step G: Preparation of intermediate compound 75a To a solution of 74a (4.00 g, 4.35 mmol) in DMF (150 mL) at room temperature, HATU (1.65 g, 4.35 mmol) was added and the reaction solution was stirred for 0.5 h. The solution was diluted with DCM (450 mL) and DIEA (1.69 g, 13.1 mmol), The mixture was then stirred at room temperature for 3 hours, and the resulting solution was quenched with water (5 mL) and concentrated under reduced pressure. The residue was purified by reversed-phase column chromatography on C18 (acetonitrile / water + 0.05% The product was purified by elution with a gradient of 0.1% TFA to give the Fmoc-protected intermediate. LCMS(ESI) C 44 H 42 Calculated for FN5O8[M+H]+: 788. 3. Measurement value 788.9.

[0216] The immediately above Fmoc-protected intermediate (200 mg, To a solution of 0.250 mmol of tetrahydrofuran (TFA), piperidine (1.25 mL) was added and the reaction solution was stirred for 1 h. The final solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography ( The amine moiety was purified by elution with a gradient of 0%-5% MeOH in DCM. LCMS(ESI) C 29 H 32 Calculated value of FN5O6[M+H]+: 5 66.2, measured value 566.3.

[0217] The amine intermediate ( A solution of BocO (2.21 g, 10.1 mmol) was added to the ) and sodium bicarbonate (1.70 g, 20.2 mmol) were added and the reaction was stirred for 3 h. The final solution was diluted with water (50 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (3×100 mL) and dried over anhydrous Na2SO4. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography ( 75a was purified by elution with a gradient of 0%-5% MeOH in DCM to give 75b. Provided by LCMS(ESI) C 34 H 40 Calculated value of FN5O8[M+H]+: 666 .3, measurement 666.5; 1 H NMR(300MHz,CD3OD) δ7.35-7 .08(m,6H),6.95-6.79(m,2H),5.01-4.91(m,1H ),4.72-4.56(m,2H),4.45-4.38(m,1H),4.45-4 .38(m,5H),3.69(s,3H),3.32-2.92(m,5H),2.1 4-1.82(m,2H),1.47(s,9H).

[0218] Step H: Preparation of intermediate compound 75b To a solution of 75a (0.665 g, 0.99 mmol) in DMF (0.5 mL), Cs 2CO3 (1.11 g, 3.40 mmol) and 3-bromoprop-1-ene (0.4 3g, 3.55mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 16 hours and 5ml of 50% saturated brine / 10% citric acid solution, then poured into ethyl acetate (2×20 ml The organic layer was washed with brine (3×20 mL) and dried over anhydrous MgSO4. The filtrate was concentrated under reduced pressure. The residue was diluted with 1%-5% MeOH in DCM and The product was purified by silica gel column chromatography eluting with acetonitrile. The fractions containing were combined and concentrated to give the title compound. ESI) C 37 H 44 FN5O8[M+H] + Calculated value: 706.3, measured value 706. 3.

[0219] Step I: Preparation of intermediate compound 76B Hydrolysis of 75b with LiOH was carried out in a similar manner as described in the preparation of intermediate 93. By following the conditions, they provided 76B.

[0220] Preparation of intermediate 77B [ka]

[0221] Step A: Preparation of Intermediate 77A (S)-1-(((9H-fluoren-9-yl)methyl)-2-(2-(phenyl)phenyl)-1-(((9H-fluoren-9-yl)methyl)-2-( ... (2-methyl-2-pyrrolidine-2-carboxylic acid) (6.16 g, 17.5 4mmol) and tert-butyl 4-(2-aminoethyl)benzylcarbamate A solution of hydrochloride (5.03 g, 17.54 mmol) was added to HATU (8.00 g, 21.0 5 mmol) and DIPEA (9.16 ml, 52.6 mmol) were added, and the reaction was then The material was allowed to warm to room temperature and stirred for 2 h. The final mixture was diluted with water and diluted with EtO The mixture was extracted with Ac, washed with brine, dried over MgSO4, and filtered. The filtrate was concentrated to give a residue. The residue was purified by column chromatography on silica gel (0-60% EtOAc in hexanes). Purification by elution with gradient elution gave 77A. MS (ESI): m / z (M+H)+ 584.5.

[0222] Step B: Preparation of Intermediate 77B To a solution of 77A (8.92 g, 15.28 mmol) in DCM (40 mL) was added HCl 4N in water (15.28 ml, 61.1 mmol) was added and the resulting solution was The mixture was stirred at room temperature overnight. The mixture was concentrated to give 77B. MS (ESI): m / z (M+H)+ 484.3.

[0223] Preparation of intermediate 86 [ka] TIFF2025041712000114.tif59166

[0224] Step A: Preparation of intermediate 77 (S)-(9H-Fluoren-9-yl)methyl 2 in DCM (140 ml) at 0° C. -((4-(aminomethyl)phenethyl)carbamoyl)-2-methylpyrrolidine-1- A solution of carboxylate hydrochloride (77B) (5.87 g, 11.29 mmol) was added to DI EA (5.91 ml, 33.9 mmol) and CBZ-Cl (1.726 ml, 11. 85mmol) was added dropwise and the resulting solution was stirred at 0°C for 4 hours. The mixture was partitioned between 100 mL of ethyl acetate and DCM (200 mL) and the aqueous phase was extracted with DCM (100 mL). The combined organic layers were dried over Na2SO4, and the residue was purified on a silica gel column with EtOAc. Purification using hexane as the elution solvent gave 77. LC / MS: (M +1) + :618.3.

[0225] Step B: Preparation of intermediate 78 77 (5.5 ml) in THF (100 ml), water (50 ml) and MeOH (30 ml) A solution of 6 g, 9.00 mmol) was added to a 1N NaOH solution (45.0 ml, 45.0 m mol) was added and the resulting solution was stirred at room temperature for 2 hours. The volatile components were evaporated and the water The solution was added to dioxane (200 ml) and Boc2O(2. The resulting mixture was stirred at 0°C to room temperature overnight. The volatile components were evaporated on a rotary evaporator and the aqueous layer was diluted with DCM (3×1 The combined organic layers were dried over Na2SO4, concentrated, and the residue was extracted with silica gel. 78 was purified on a gel column using EtOAc / hexane as the eluent to give 79. LC / MS: (M+1) + :496.2.

[0226] Step C: Preparation of Intermediate 79 A solution of 78 (4.04 g, 8.15 mmol) in MeOH (100 ml) was added at 10% Pd / C (0.867 g, 0.815 mmol) was added and the resulting mixture was heated under a H balloon. The mixture was subjected to hydrogenation at room temperature for 1.5 hours through celite. Concentration gave 79. LC / MS: (M+1) + :362.2.

[0227] Step D: Preparation of Intermediate 80 To a solution of 79 (2.58 g, 7.14 mmol) in DMF (15 ml), -en-1-yl 4-methylbenzenesulfonate (0.858g, 3.57mmol ) and K2CO3 (1.973 g, 14.27 mmol) were added, and the resulting mixture was stirred for 8 The mixture was heated at 0° C. for 6 h. After cooling to room temperature, the mixture was filtered and the filtrate was purified by reverse phase MPLC. Acetonitrile (0.05% TFA) / water (0.05% TFA) was used as the elution solvent. The product was purified as a TFA salt, which was further dissolved in DCM (100 mL). The aqueous layer was partitioned between 1N NaOH (50 mL) and 1N NaOH (aq.). The aqueous layer was further diluted with DCM (2×5 The combined organic layers were dried over Na2SO4 and concentrated to give 80. LC / MS: (M+1) + :430.3.

[0228] Step E: Preparation of Intermediate 81 To a solution of 80 (0.95 g, 2.211 mmol) in DMF (15 ml), 4-oxobutanoic acid (0.321 g, 2.433 mmol), HATU (1.00 9g, 2.65mmol) and DIEA (0.927ml, 5.31mmol) were added. The resulting solution was stirred at room temperature for 1 h. The solution was diluted with EtOAc (200 mL) and brine ( 100 mL), and the organic layer was washed with brine (2×100 mL) and Dry over O4, concentrate, and purify the residue onto a silica gel column eluted with EtOAc / hexane. Purification using ethyl acetate as a starting material gave 81. LC / MS: (M+1) + :544.2.

[0229] Step F: Preparation of Intermediate 82 To a solution of 81 (1.165 g, 2.143 mmol) in DCM (12 ml), HCl (4N in dioxane) (5.36 ml, 21.43 mmol) was added. The resulting solution was After stirring at room temperature for 3 h, the mixture was concentrated to give 82. LC / MS: (M+1) + :444.2.

[0230] Step G: Preparation of Intermediate 83 To a solution of 82 (1.003 g, 2.089 mmol) in DMF (20 ml), Fmo cL-Tyr(Me)-OH (0.959g, 2.298mmol), HATU(0. 914g, 2.403mmol) and DIEA (1.095ml, 6.27mmol). The solution was added and stirred at room temperature for 50 min. The solution was diluted with EtOAc (200 mL) and The organic layer was washed with brine (2×100 mL) and partitioned between the column and the line (100 mL). The combined organic layers were dried over Na2SO4, concentrated, and the residue was purified by silica gel column with EtO Purification using Ac / hexane as the elution solvent gave 83. LC / MS: M+1) + :843.4.

[0231] Step H: Preparation of Intermediate 84 To a solution of 83 (1.63 g, 1.934 mmol) in acetonitrile (10 ml), Piperidine (0.574 ml, 5.80 mmol) was added and the resulting solution was incubated at room temperature for 1 h. The residue was resuspended in acetonitrile (20 mL) and concentrated again. This cycle was repeated once and the residue was further dried under high vacuum to give 84. LC / MS: (M+1) + :621.3.

[0232] Step I: Preparation of intermediate 85 To a solution of 84 (1.2 g, 1.933 mmol) in DMF (15 ml) was added Fmoc- L-Thr(tBu)-OH(0.922g, 2.320mmol), HATU(0.9 (19 g, 2.416 mmol) and DIEA (0.844 ml, 4.83 mmol). The resulting solution was stirred at room temperature for 1 h. The solution was diluted with EtOAc (200 mL) and The organic layer was washed with brine (2×100 mL) and then with NaCl. Dry over 2SO4, concentrate, and purify the residue onto a silica gel column eluted with EtOAc / hexane. Purification using hexane as solvent gave 85. LC / MS: (M+1) + :1000 .2.

[0233] Step J: Preparation of Intermediate 86 The 85 (1.94 g, 1 To a solution of 0.940 mmol of tetrahydrofuran, piperidine (0.960 ml, 9.70 mmol) was added. The resulting solution was stirred at room temperature for 30 min and then concentrated. The residue was diluted with DCM / acetonitrile The mixture was then redissolved in ethanol (1:1, 20 mL) and concentrated again, and this cycle was repeated once. The residue was dried under high vacuum to give 86. LC / MS: (M+1) + :77 8.3.

[0234] Preparation of intermediate 88 [ka]

[0235] Step A - Synthesis of Intermediate 87 2-Chloro-2-chlorotrityl resin 1-1.5mmol / g (7.0g, 1-1.5 To the resin (mmol / g) was added anhydrous DCM (45 ml). The resin was shaken for 20 min, followed by DC Half amount of DIPEA 0.17N in M ​​(3.67 ml, 21.00 mmol), Fmoc -D-Dap(Boc)-OH (3.28 g, 7.70 mmol) was then added to the remaining DC DIPEA 0.17N in M ​​(3.67 ml, 21.00 mmol) was added. The resin was then shaken overnight at room temperature, rinsed with DCM and dried. 5% DIPEA and 10% MeOH, shaken for 2 hours, then filtered, Rinse with DCM (3x), DMF (3x) and DCM (3x) then dry under vacuum. Allowing to stand gave resin 87 which was used as is in the next step.

[0236] Step B - Synthesis of Intermediate 88 Resin 87 (4.5 g, 2.475 mmol) was dissolved in 5% piperidine in DMF (30 ml). The product was manually deprotected with 5% piperidine in DMF (30 ml) for 30 min, filtered, and deprotected with 5% piperidine in DMF (30 ml). The resin was then reprocessed for an additional 30 minutes, filtered, and then rinsed with DMF and DCM and dried. The oil was then dissolved in DMF (30 ml) and diluted with Fmoc-Ala-OH (1.541 g, 4.95 ml). mol), HATU (1.694 g, 4.46 mmol) and DIPEA (1.729 ml, 9.90 mmol) for 2 hours, then filtered and diluted with DMF The resin was then dissolved in 10 mL of DCM (60 ml) and rinsed with DCM and then dried. % AcOH and TFE for 90 min, filtered, and the filtrate was concentrated to give 88. LC / MS: [2M+H] + =995.01.

[0237] Example 1A - Alternative synthesis of Ex-01 and preparation of Ex-25 from this compound: Compound Ex-01 presented above may alternatively be prepared according to the following scheme: The product Ex-25 can be prepared from Ex-01: [ka] TIFF2025041712000117.tif231166TIFF2025041712000118.tif199163TIFF2025041712000119.tif202169

[0238] Step A - Synthesis of intermediate Int-cd1 Int-3c (synthesized from intermediate 107 below) (7.0 A solution of Int-2d (3.63 g, 9.84 mmol, (prepared as follows) and HATU (3.74 g, 9.84 mmol) were added, followed by DMF DIPEA (6.87 ml, 39.4 mmol) was added and the mixture was left to stand. The mixture was warmed to room temperature and stirred for 1 h. The mixture was quenched with brine at 0° C. and diluted with EtOAc. The combined organic fractions were washed with brine, dried over MgSO4, and filtered. The residue was purified by column chromatography on silica gel (petroleum ether The Int- cd1 was given. LC / MS: [M+1]+=1000.5.

[0239] Step B - Synthesis of intermediate Int-cd2 of Int-cd1 (3.48 g, 3.48 mmol) in acetonitrile (50 ml) To the solution, piperidine (1.72 ml, 17.40 mmol) was added, and the resulting solution was cooled to room temperature. The mixture was stirred at rt for 3 h. The mixture was concentrated and the residue was diluted with DCM / acetonitrile (1:1, 20 mL ), concentrated again, and the residue was dried under vacuum to obtain crude Int-cd2. The product was obtained as a product. LC / MS: (M+1) + =778.5.

[0240] Step C - Synthesis of intermediate Int-cd3 76 (preparation shown in Example 1 above) (2.45 g) in DMF (70 ml) at 0° C. , 3.01 mmol) and Int-cd2 (2.69 g, 3.46 mmol) , HATU (1.37 g, 3.61 mmol) was added, followed by DIEA (1.05 ml, 6.02 mmol) was added. The resulting solution was stirred at room temperature for 50 min and then added with EtOAc. (500 mL) and brine (200 mL). The organic layer was partitioned between brine (2×2 00 mL), dried over Na2SO4, concentrated, and the residue was subjected to column chromatography on silica gel. Purification was performed by chromatography (eluting with a gradient of 1%-5% MeOH in DCM). This gave Int-cd3. LC / MS: (M+1) + =1574.7.

[0241] Step D - Synthesis of intermediate Int-cd4 Int-cd3 (1.91 g, 1 A room temperature solution of 1.21 mmol) was bubbled with N2 for 30 min, followed by Zhan catalyst-1B (0.445 g, 0.607 mmol) was added. The resulting mixture was further stirred at room temperature for 30 The mixture was bubbled with N2 for 1 min and then heated at 55° C. for 5 h. After cooling to room temperature, the mixture was mixed. The mixture was filtered through Celite, the filtrate was concentrated, and the residue was purified by column chromatography on silica gel. (eluted with a gradient of 1%-5% MeOH in DCM) to give In gave t-cd4 (as a mixture of cis and trans olefins). LC / MS: (M+1) + =1546.8.

[0242] Step E - Synthesis of intermediate Int-cd5 Int-cd4(systole) in DCM (20 ml) and acetonitrile (50 ml) A solution of 5.49 g (3.55 mmol) of olefin (mixture of fin and trans olefins) was added to Then piperidine (1.76 ml, 17.8 mmol) was added. The resulting solution was stirred at room temperature for 2 hours. The mixture was stirred for 10 min, then concentrated, and the residue was suspended in acetonitrile (20 ml) and concentrated again. The residue was then dried under vacuum to give Int-cd5 (cis olefin and tra The crude mixture was obtained as a mixture of acetonitrile and olefins. LC / MS: (M+1) + = 1323.8.

[0243] Step F - Synthesis of intermediate Int-cd6 Int-cd5 (cis and trans olefins) in DMF (70 ml) at 0°C Int-1d (4.70 g, 3.55 mmol) and Int-1d (2.21 g, 4 HATU (1.76 g, 4.62 mmol) and and DIEA (1.55 ml, 8.88 mmol) were added. The resulting solution was allowed to warm to room temperature. The mixture was stirred for 1 h, then diluted with EtOAc (300 mL) and brine (200 mL). The aqueous layer was extracted with EtOAc (200 mL) and the EtOAc layers were combined and washed with brine. The residue was purified by washing with hexane (3×200 mL), drying over Na2SO4, and concentrating, and the residue was purified by column chromatography on silica gel. Column chromatography (eluted with a gradient of 1%-5% MeOH in DCM) By purifying it, Int-cd6 (a mixture of cis and trans olefins) was obtained. LC / MS: (M+1) + =1802.8

[0244] Step G-Synthesis of intermediate Int-cd7 Int in THF (100 ml), MeOH (30 ml) and water (30 ml) at 0° C. -cd6 (as a mixture of cis and trans olefins) (5.41 g, 3 0.00mmol) in 1N LiOH solution (24.0ml, 24.0mmol) was added dropwise and the resulting solution was stirred at 0° C. for 3 h. The mixture was diluted with 1N HCl at 0° C. The volatile components were evaporated and the aqueous layer was brought to pH 5 with 1N HCl. The mixture was then frozen and lyophilized, and the residue was purified by column chromatography on C18. Fee (Acetonitrile (0.05% TFA) / Water (0.05% TFA) Gradient Int-cd7 (cis and trans olefins) was purified by (as a mixture of fins) as the TFA salt. Acetonitrile (750 mL) and The thus obtained Int-cd7 TFA salt (cis olefin and and trans olefins) was added to 150 ml of 0.1N HCl solution at 0°C. 1, 15.00 mmol) was added dropwise, and the resulting solution was then stirred at 0° C. for 5 minutes. Freezing and lyophilization gave Int-cd7 as the HCl salt (cis olefin and as a mixture of trans olefins). LC / MS: (M+1) + =1566.6 .

[0245] Step H - Synthesis of intermediate Int-cd8 Int from the previous step in DMF (50 ml) and DCM (1300 ml) A solution of -cd7 HCl salt (1.01 g, 0.630 mmol) was added to DIEA (0.33 0ml, 1.890mmol) and HATU (0.287g, 0.756mmol) The resulting solution was stirred at room temperature for 2 h, the volatile components were evaporated and the residue was concentrated in EtOAc. (400 mL) and brine (200 mL). The aqueous layer was diluted with EtOAc (30 0 mL) and the combined organic layers were washed with brine (3 × 100 mL) and NaSO The residue was purified by column chromatography on silica gel (1% in DCM) -10% MeOH gradient) to obtain Int-cd8 ( LC / MS: (M+ 1) +=1548.8.

[0246] Step I - Synthesis of intermediate Int-cd9 Int-cd8 (1.2 ml) obtained in the previous step in MeOH (100 ml) 2 g, 0.788 mmol) in a solution of 10% Pd / C (0.645 g, 0.607 mm ol) was added and the resulting mixture was subjected to hydrogenation via a H2 balloon at ambient temperature for 7 h. After 7 hours, the reaction was filtered over Celite, the filtrate was concentrated, and the residue was purified by column chromatography on silica gel. Column chromatography (eluting with a gradient of 1%-10% MeOH in DCM) Purification by HPLC gave Int-cd9. LC / MS: (M+1) + =1550. 9.

[0247] Step J - Synthesis of Compound Ex-01 as HCl Salt To a solution of Int-cd9 (1.14 g, 0.735 mmol) in DCM (6 ml), TFA (12 ml, 156 mmol) was added and the resulting solution was stirred at ambient temperature for 30 min. The mixture was then concentrated and the residue was dissolved in DCM (20 mL) and toluene (20 mL). The resulting mixture was concentrated, and the residue was redissolved in DCM (20 mL) and HCl ( The resulting mixture was treated with 4N in dioxane (0.919 ml, 3.68 mmol). The mixture was concentrated to give the product as a solid. The solid product was dissolved in acetonitrile (2 The solution was then redissolved in 100 mL of 1N HCl and water (100 mL). The resulting solution was incubated at 0° C. for 2 min. Stirring, followed by freezing and lyophilization gave Ex-01 as the HCl salt. / MS:(M+1) + =1394.7.

[0248] Step K - Synthesis of Example Ex-25 as TFA Salt Ex-01 HCl salt (870 mg) in DMF (1.2 ml) and water (0.6 ml) , 0.608 mmol) and Int-4b (170 mg, 0.669 mmol, prepared A solution of HATU (254 mg, 0.669 mmol) and DIEA (425 The resulting solution was stirred at room temperature for 1 hour, then 1. The mixture was quenched by the addition of 2 mL of water. The mixture was filtered and the filtrate was purified by column chromatography on C18. Chromatography (acetonitrile (0.05% TFA) / water (0.05% TFA) Purification by LC / HPLC (eluting with 100% ethyl acetate) gave Ex-25 as a TFA salt. MS:M + =1550.6.

[0249] Preparation of Example Ex-25 as Step L-Cl Salt Two columns were loaded with 73.6 g of AG MP-1 ion exchange resin chloride form (cat# 141-1841 BIO-RAD) so that the total resin content in each column was 36.8 g. Each column was washed with water (2×80 ml), followed by 20% acetonitrile in water. The mixture was washed with 20% acetonitrile in water (100 ml). Ex-25 TFA salt (737 mg, 0.443 mmol) prepared in step The solution was loaded equally onto two resin columns, and then each column was washed with 20% acetonide in water. The eluates were combined, frozen, and lyophilized to obtain Ex -25 was given as the chloride salt. LC / MS: + =1550.6.

[0250] The following are useful in the synthesis of Ex-01 and Ex-25 just described: Description of some intermediates.

[0251] Preparation of intermediate Int-1d The intermediate compound Int-1d was prepared from starting materials according to the following scheme. [ka]

[0252] Step A - Synthesis of Int-1da D-Dap(Boc)-OMe HCl salt (4.10 g) in DMF (40 ml) at 0° C. , 16.10mmol), Fmoc-Ala-OH (5.01g, 16.10mmol) and HATU (6.43 g, 16.90 mmol) in a solution of DIPEA (7.03 m 1, 40.2 mmol) was added, and the mixture was stirred at 0°C for 2 hours and then left to stand overnight in a refrigerator. The mixture was quenched with water at room temperature and extracted with EtOAc. The fraction was washed with half brine, dried over Na2SO4, and filtered. The residue was purified by column chromatography on silica gel (hexanes / Purification by elution with a gradient of EtOAc gave Int-1da. LC / MS: [M+H]+=512.3.

[0253] Step B - Synthesis of Int-1d Int-1da (8 0.03 g, 15.70 mmol) and 0.8 N calcium chloride (19.62 ml, 15 A solution of 0.691 g (17.27 mmol) of solid sodium hydroxide (0.70 mmol) was added to the l) was added. The mixture was stirred at room temperature overnight. The mixture was concentrated and the pH was adjusted to about 2 with 0.5N It was acidified (ca. 40 mL), extracted twice with EtOAc, washed with brine, and dried over Na2SO4. The residue was purified by column chromatography on C18 (acetonitrile / water + 0.1% TFA gradient) to give Int-1d. LC / MS: [M+H]+=498.25.

[0254] Preparation of intermediate Int-1d for use in the preparation of Ex-01 and Ex-25 This part of the molecule is responsible for converting small peptide rings into larger peptide rings. For example, but not limited to, Da Varying the spacers used in the synthesis, including using p- and D-Ala Other similar "linkers" may be used in place of Int-1d depending on the above.

[0255] Preparation of intermediate Int-2d Intermediate Int-2d, useful as a "linker" in the preparation of compounds of the invention, is shown below: It was prepared according to the scheme: [ka]

[0256] Step A - Synthesis of Int-2da 4-Bromobenzaldehyde ( 15.00g, 81mmol), potassium tert-butyl N-[2-(trifluoro Boranediyl)ethyl]carbamate (20.97 g, 84 mmol), cesium carbonate ( 52.8 g, 162 mmol) and 1,1'-bis(diphenylphosphino)ferrocene Palladium(II) dichloride dichloromethane complex (Pd(II)(dppf)C A solution of l2 (1.99 g, 2.43 mmol) was warmed to 76° C. and stirred overnight. The mixture was quenched with half-saturated aqueous ammonium chloride solution at room temperature and extracted with EtOAc. The combined organic fractions were washed with brine, dried over Na2SO44, filtered and concentrated in vacuo The residue was purified by column chromatography on silica gel (Dimethyl ether / Et0Ac, 100% aqueous solution). Purification by HPLC (eluted with gradient elution) gave Int-2da. LC / MS: M-56+1) + =193.0.

[0257] Step B - Synthesis of Int-2db Int-2d in DCM (120 ml) and AcOH (3 ml) at room temperature in a water bath a (12.9 g, 51.7 mmol) and pent-4-en-1-amine (6.61 g , 78 mmol) was added to a solution of sodium triacetoxyborate (32.9 g, 15 5 mmol) was added in portions and the mixture was stirred for 30 min. The reaction was diluted with 3 ml of water at 0° C. Quench more slowly, pour into 1N NaOH (500 mL), stir for 15 min, then The mixture was extracted with DCM, dried over Na2SO4 and concentrated. The residue was purified by column chromatography on silica gel. Purification by chromatography (eluting with a gradient of DCM / MeOH) gave Int-2db. LC / MS: (M+1) + =319.2.

[0258] Step C-Synthesis of Int-2dc Int-2db (8.48 g, 20.77 mmol) and 4 in DMF (40 ml) To a solution of 10-methoxy-4-oxobutanoic acid (3.02 g, 22.85 mmol), U (9.48 g, 24.92 mmol) and DIPEA (8.71 ml, 49.8 mm ol) was added. The resulting solution was stirred at room temperature for 1 h and then saturated aqueous NaHCO3 ( The mixture was quenched with EtOAc (500 mL) and saturated aqueous NaHCO3 (10 mL). (200 mL), and the organic layer was washed with brine (3×200 mL) and The residue was purified by silica gel column (hexanes / EtOAc elution) and dried over 2SO4 and concentrated. Purification by HPLC (eluted with 100 mL of ethyl acetate) gave Int-2dc. LC / MS: (M+1) + =433.4.

[0259] Step D-Int-2d synthesis To a solution of Int-2dc (2.9 g, 6.70 mmol) in DCM (15 mL), Hot 4M HCl in dioxane (10 mL) was added. The reaction mixture was stirred at room temperature for 1 h. The mixture was concentrated under reduced pressure to give methyl 4-((4-(2-aminoethyl)benzene (pent-4-en-1-yl)amino)-4-oxobutanoate hydrochloride (I nt-2d) was obtained. LC / MS [MHCl+H] + =333.3.

[0260] Preparation of Int-3c from 107 used in the synthesis of Ex-01 and Ex-25 above Intermediate Int-3c was prepared according to the following scheme: [ka]

[0261] Step A - Synthesis of Int-3ca 107 (the preparation of which is given in the synthesis of 109 and will be described later) in THF (100 ml) at room temperature 116, this intermediate compound is Ex-53, E (used in the synthesis of Ex-54 and Ex-55) (10.34 g, 21.65 mm ol) was added 2N lithium hydroxide monohydrate (43.3 ml, 87 mmol), The mixture was warmed to 45 °C and stirred overnight to give Int-3ca as a crude solution. LC / MS: (M+1) + = 464.3. The reaction mixture was cooled to 0°C and 1M It was treated with HCl (40 mL) and the mixture was used directly for the next step.

[0262] Step B - Synthesis of Int-3c The crude Int-3ca prepared in the previous step was diluted with NaHCO3 (1.725 g, 20.54 mmol) and Fmoc-OSu (3.81 g, 11.30 mmol). was added at 0° C. The reaction mixture was stirred at 0° C. for 2 h and then treated with 1 M HCl (20.5 mL). The combined organic layers were washed with brine (2×1 00 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The mixture was purified by silica gel column chromatography using 2% to 5% MeOH in DCM. Purification by elution with 20 ml of diethyl ether gave Int-3c. LC / MS: (M+1) + =686.4.

[0263] Preparation of intermediate Int-4b Intermediate Int-4b was prepared according to the following scheme: [ka]

[0264] Step A - tert-Butyl-3-(2-hydroxyethoxy)propionate (pro Synthesis of Int-4ba from ponate tert-Butyl 3-(2-hydroxyethoxy)propanoate in DCM (2 mL) A solution of CBr4 (1395 mg, 4.2 mmol) was added to a solution of 1,2-dichloro- ... 1 mmol) and PPh3 (965 mg, 3.68 mmol) were added at 0 °C. The mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography. Chromatographically, the solution was purified using a gradient of 1%-15% ethyl acetate in petroleum ether. The fractions containing the desired product were combined and concentrated to give t The acetonitrile thus prepared gave ert-butyl 6-bromohexanoate. tert-Butyl 6-bromohexanoate (5 g, 19.91 m A solution of 1.2 mmol) was treated with trimethylamine (13.56 mL, 59.7 mmol), The resulting solution was heated at 50° C. overnight. The solution was concentrated to give Int-4ba. LC / MS:M + =230.3.

[0265] Step B - Synthesis of Int-4b To a solution of Int-4ba (6.8 g, 21.92 mmol) in DCM (6 ml), 4N HCl in oxane (27.4 ml, 110 mmol) was added, and the resulting solution was The mixture was then concentrated to give Int-4b. LC / M S:M + =174.3.

[0266] Preparation of intermediate Int-2d for use in the preparation of Ex-01 and Ex-25 This part of the molecule is R 1 , R 2 and R 8 Small with substituents It can be described as a "linker" that cyclizes the peptide ring. Other similar "linkers" are The following can be used in place of nt-2d to prepare the examples of the invention described herein: 1 is a description of other "linkers" that can be used to

[0267] Preparation of intermediate Int-2e Intermediate Int-2e, useful as a "linker" in the preparation of compounds of the invention, is shown below: It was prepared according to the scheme: [ka]

[0268] Step A - Synthesis of intermediate Int-2ea tert-Butyl (2-(3-oxoisoindoline-5-yl)phenyl)acetate in DCE (20 mL) A solution of 1.60 g (5.79 mmol) of NsCl (1. 93g, 8.69mmol), triethylamine (1.76g, 17.4mmol) and and DMAP (0.141 g, 1.16 mmol) were added. The reaction mixture was heated at 40° C. for 14 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel. The product was purified by chromatography (eluting with a 1%-40% gradient of EtOAc in PE). The resulting mixture was purified by HPLC to give Int-2ea. LC / MS: (M+Na) + :=484.4.

[0269] Step B - Synthesis of intermediate complex Int-2eb Int-2ea (11.3 g, 24 To a solution of 1.001 g (0.05 mmol) of 1,000 sulphate was added LiOH (1.76 g, 73.5 mmol). The mixture was stirred at 25° C. for 5 h, and then the resulting solution was adjusted to pH 4-5 with HCl (1M). The solution was extracted with EtOAc, and the combined organic layers were washed with brine and anhydrous NaSO. The mixture was dried at 4°C and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel. Purify by chromatography (eluting with a gradient of 1%-6% MeOH in DCM). This gave Int-2eb. LC / MS: (M+Na) + :=502.2.

[0270] Step C - Synthesis of intermediate Int-2ec A solution of Int-2eb (1.70 g, 3.55 mmol) in THF (8 mL) was Ran (0.147 g, 10.6 mmol) was added at 0° C. The reaction mixture was incubated at 25° C. for 14 h. The mixture was stirred for 1 h, and then the resulting solution was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel. Purify by column chromatography (eluting with a gradient of 1%-50% EtOAc in PE). This gave Int-2ec. LC / MS: (M+NH4] + =483.2.

[0271] Step D - Synthesis of intermediate Int-2ed To a solution of Int-2ec (4.50 g, 9.67 mmol) in DMF (150 mL) , K2CO3 (2.01 g, 14.5 mmol) and 3-bromoprop-1-ene (1 The reaction mixture was stirred at room temperature for 5 hours and then diluted with water. The mixture was diluted and extracted with EtOAc. The combined organic layers were washed with brine and anhydrous Na2SO4 The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel. Purify by chromatography (eluting with a gradient of 1%-50% EtOAc in PE). This gave Int-2ed. LC / MS: (M+H) + :=506.2.

[0272] Step E - Synthesis of intermediate Int-2e To a solution of Int-2ed (4.50 g, 8.90 mmol) in DMF (35 mL), DBU (1.35 g, 8.90 mmol) and 2-mercaptoethanol (2.08 g The reaction mixture was stirred at room temperature for 14 hours and then cooled to 5°C. Column chromatography (column: 330 g; mobile phase A: water / 0.05% TFA, mobile phase B: water / 0.05% TFA, Phase B: ACN; flow rate: 85 mL / min; gradient: 10% B to 20% B in 15 min 20%B to 45%B in 15 minutes (detector: UV 210 nm; Rt = 20 minutes) Purification of the product gave Int-2e. LC / MS: (M+H) + :=321.2. 1 H NMR(300MHz,CDCl3) δ7.21-7.13(m,2H),7 .10-7.01(m,1H),5.96-5.79(m,1H),5.30-5.09 (m,2H),4.58(s,2H),3.84(s,2H),3.43-3.19(m ,4H),2.76(t,J=7.1Hz,2H),1.41(s,9H).

[0273] Preparation of intermediate Int-2f-1 Intermediate Int-2f-1, useful as a "linker" in the preparation of compounds of the present invention, It was prepared according to the following scheme: [ka] TIFF2025041712000126.tif45158

[0274] Step A - Synthesis of intermediate Int-2fa 4-Bromobenzaldehyde (20.0 g, 108 mmol) in DCM (225 mL) ), (S)-2-methylpropane-2-sulfinamide (12.5 g, 103 mmol ), MgSO4 (130 g, 1081 mmol), pyridine 4-methylbenzene Sulfonate (1.35 g, 5.40 mmol) was added under nitrogen protection. This mixture was mixed for 2 After stirring at 5° C. for 72 hours, the resulting solution was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (1%-15% EtOAc in PE). Purification by HPLC (eluted with gradient elution) gave Int-2fa. LC / MS: M+H) + :=287.9, 289.9.

[0275] Step B - Synthesis of intermediate Int-2fb (racemic form) and enantiomer Int- Separation into 2fb-1 and Int-2fb-2 Dissolution of Int-2fa (20.0 g, 65.9 mmol) in anhydrous DCM (200 mL) Add but-3-en-1-yl magnesium bromide (15.7 g, 99 mmol) to the solution. The mixture was stirred at -48°C for 2 hours and then cooled to room temperature. The mixture was quenched with aqueous NH4Cl (400 mL) and extracted with DCM. The mixture was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give The collected residue (containing the Int-2fb racemic mixture) was subjected to column chromatography on silica gel. Purify by column chromatography (eluting with a gradient of 1%-35% EtOAc in PE). This gave Int-2fb-1 and Int-2fb-2. LC / MS: (M+H ) + :=344.0,346.0.

[0276] Step C - Synthesis of intermediate Int-2fc-1 Int-2fb- was added to a solution of HCl (100 mL, 4N in 1,4-dioxane) at room temperature. 1 (17.0 g, 46.9 mmol) was added. The reaction solution was stirred for 1 h and then cooled under reduced pressure. Concentration with HCl gave Int-2fc-1. LC / MS: (M+H-HCl) + : =240.0,242.0.

[0277] Step D - Synthesis of intermediate Int-2fd-1 Int-2fc-1 (8.20 g, 28.2 m) in 1,4-dioxane (200 mL) A solution of Teoc-OSu (8.03 g, 31.0 mmol) and Teoc-OSu (8.03 g, 31.0 mmol) was added with TEA ( 8.55 g, 84 mmol) was added at 25° C. The mixture was stirred for 2 hours and then diluted with water. The mixture was quenched and extracted with petroleum ether (PE). The combined organic layers were concentrated under reduced pressure to give a residue. The mixture was purified by column chromatography on silica gel (1%-10% EtOAc in PE gradient). Purification by HPLC (eluted with 1000 mL of ethyl acetate) gave Int-2fd-1. LC / MS: (M +Na+CH3CN) + :=447.3, 449.3.

[0278] Step E - Synthesis of intermediate Int-2fe-1 Int-2fd-1 (15.1 g, 37.3mmol), potassium (2-((tert-butoxycarbonyl)amino)ethyl Cs2CO3(36.5 g, 112 mmol) was added to a solution of PdCl2(dppf) (1.37 g, 1.87 mmol). l) was added under nitrogen protection. The mixture was stirred at 80°C for 40 hours. The resulting solution was washed with water. The mixture was quenched and extracted with EtOAc. The combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (1% in PE). -40% EtOAc gradient) to give Int-2fe- 1 was obtained. LC / MS: (M+Na) + :=471.4.

[0279] Step F - Synthesis of intermediate Int-2f-1 A solution of Int-2fe1 (10.6 g, 22.4 mmol) in THF (100 mL) To this mixture was added 1N TBAF in THF (44.9 mL, 44.9 mmol). The mixture was stirred at room temperature for 16 h, then quenched with water and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (1%-70% EtOAc in PE). elution with a gradient of 0.01 to 0.1%, followed by column chromatography on C18 (column :330g; Mobile phase A: Water (10mm NH4HCO3), Mobile phase B: ACN; Flow rate: 8 0 mL / min; Gradient: 10% B to 10% B in 10 min, 20% B to 45% B 10 min to 45%B, 20 min to 70%B, detector: UV 210 nm; Rt=2 5 min) to give Int-2f-1. LC / MS: (M+H) + :=305.1. 1 H NMR(300MHz,CD3OD) δ7.27-7.1 6(m,4H),5.85-5.75(m,1H),5.00-4.85(m,2H), 3.79(t,J=7.0Hz,1H),3.32-3.21(m,2H),2.75( t,J=7.4Hz,2H),2.98-1.72(m,4H),1.42(s,9H) .

[0280] Example 2 Preparation of Ex-50 and Ex-52 [ka]

[0281] Compound Ex-50 can be prepared according to the scheme below, the preparation of which is described in Example 1 herein. This is prepared from the compound Ex-01 under suitable conditions according to the following scheme: It is prepared by reacting with intermediate Int32, [ka]

[0282] Step A: Preparation of intermediate Int-32A tert-Butyl 3-(2-(2-bromoethoxy)phenylacetate in acetonitrile (10 ml) )ethoxy)propanoate (5 g, 16.82 mmol) in a solution of trimethylamine (33% in ethanol, 11.46 ml, 50.5 mmol) was added and the resulting solution was stirred for 5 The mixture was heated at 0° C. overnight. The solution was concentrated to give 2-(2-(3-(tert-butoxy) -3-Oxopropoxy)ethoxy)-N,N,N-trimethylethanaminium bromide LC / MS: (M) + :276.5.

[0283] Step B: Preparation of intermediate Int-32 2-(2-(3-(tert-butoxy)-3-oxopropionyl)) in DCM (20 ml) (oxy)ethoxy)-N,N,N-trimethylethanaminium bromide (Int-32A ) (5.99 g, 16.81 mmol) was added to a solution of HCl (4N in dioxane) (21. 01 ml, 84 mmol) was added and the resulting solution was stirred at room temperature overnight. The solution was concentrated. This resulted in 2-(2-(2-carboxyethoxy)ethoxy)-N,N,N-trimethylethoxy To give tantaminium bromide (Int-32). LC / MS: (M) + :220.1 .

[0284] Preparation of Example Compound Ex-50 [ka]

[0285] Ex-01 (crude product) (17.4 mg, 0.012 mmol) in DMF (2 ml) and 2-(2-(2-carboxyethoxy)ethoxy)-N,N,N-trimethylethoxy To a solution of ammonium bromide (Int-32) (4.49 mg, 0.015 mmol) , HATU (5.69 mg, 0.015 mmol) and DIEA (6.54 μl, 0. 037 mmol) was added and the resulting solution was stirred at room temperature for 50 min and then subjected to reverse phase HPLC. Therefore, purification was performed using acetonitrile (0.1% formic acid) / water (0.1% formic acid) as the mobile phase. This gave Ex-50. LC / MS: + =1596.3.

[0286] Preparation of Example Compound Ex-52 Compound Ex-52 was prepared in a manner similar to that for compound Ex-50, except that the amount of Ex-01 was Ex-51 was used instead. Ex-52 was prepared according to the method described herein. The product was purified by reverse phase HPLC. LC / MS: M+=1593.8.

[0287] Example 3 Preparation of Ex-53, Ex-54 and Ex-55 [ka] TIFF2025041712000131.tif77146

[0288] Compounds Ex-53, Ex-54 and Ex-55 were prepared in a similar manner to the above compounds. Prepared from intermediate 115 (prepared below) according to the following scheme and synthetic description: [ka]

[0289] Step A - Synthesis of Intermediate 116 115 in DMF (1.5 ml), DCM (10 ml) and water (0.5 ml) at 0 °C (66.3 mg, 0.044 mmol) was added to a solution of DIPEA (0.030 ml, 0.1 73 mmol) was added, followed by HATU (18.50 mg, 0.049 mmol). The mixture was stirred for 30 min. The mixture was concentrated in vacuo and purified by column chromatography on C18. Fee (30 g, acetonitrile + 0.05% TFA / water + 0.05% TFA 90:1 0 to 40:60), 116 was purified directly into the E and Z isomers. as a mixture of isomers, as well as 116 as a pure fraction of the E or Z isomer LC / MS (major isomers) LC / MS:M + =1481.19;LC / MS (minor isomer): LC / MS: M + =1480.

[0290] Step B - Synthesis of compound Ex-54 116 (25.1 mg, 0.017 mmol) and Pd- A solution of C10% (3.61 mg, 3.39 μmol) was hydrogenated at 1 atm for 1 h. The reaction was filtered over celite and concentrated. The residue was treated with DCM / TFA 1:1 for 30 min. This was then concentrated, treated with 4N HCl in dioxane (100 uL), and concentrated. This afforded Ex-54 in the form of an HCl salt. + =1383.44 .

[0291] Step C - Synthesis of compound Ex-55 Example compound Ex-55 was prepared in the form of a formate salt from Ex-54, by synthesis of compound Ex-50. The synthesis was performed in the same manner as described in Example 2. LC / MS: + =1 583.69.

[0292] Step D - Synthesis of compound Ex-53 Example compound Ex-53 was prepared in the form of an HCl salt from intermediate compound 116 to compound Ex -51 was prepared in the same manner as described in Example 1. LC / MS :M + =1381.33.

[0293] Compounds Ex-53, Ex-54 and Ex-55 were finally synthesized from intermediate compound 115. However, the preparation of the intermediates necessary to provide intermediate compound 115 can be carried out by the method of The preparation of 103 is described in the schemes and syntheses below. [ka]

[0294] Step A - Synthesis of Intermediate 100 4-Bromo-2-hydroxybenzoate in degassed toluene (45 ml) and water (15 ml) Zucaldehyde (3.00 g, 14.92 mmol), potassium tert-butyl N-[ 2-Trifluoroboraniudyl ethyl] Carbamate (3.82 g, 15.22 mmol), cesium carbonate (17.02 g, 52 .2mmol) and 1,1'-bis(diphenylphosphino)ferrocene-palladium (ii) Solution of dichloride dichloromethane complex (0.611 g, 0.746 mmol) The mixture was heated to 75° C. and stirred overnight. The mixture was quenched with and extracted with EtOAc. The combined organic fractions were washed with brine. The residue was purified by column chromatography on silica gel. By chromatography (elution with hexanes / EtOAc 99:1 to 60:40), and purification to give 100. LC / MS: (M-55) + =210.25.

[0295] Step B - Synthesis of Intermediate 101 100 (1.70 g, 6.41 mmol) and aryl bromide in DMF (10 ml) at room temperature A solution of 1.328 g (9 0.61 mmol) was added and the mixture was warmed to 50° C. and stirred for 1 h. The mixture was allowed to stand at room temperature for half The mixture was quenched with saturated aqueous sodium bicarbonate and extracted with EtOAc. The organic fraction was washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel (hexanes / EtOAc 99: 1 to 70:30) to give 101. LC / MS: (M -55) + =250.29.

[0296] Step C - Synthesis of Intermediate 102 101 (1.76 g) prepared in the previous step in MeOH (100 ml) at room temperature , 5.76 mmol), 4 A molecular sieves (2 g) and ammonium acetate (4 0.44 g, 57.6 mmol) of sodium cyanoborohydride (0.380 g (6.05 mmol) was added and the mixture was shaken overnight. The mixture was concentrated and washed with water at room temperature. The mixture was quenched with 100 ml of ethyl acetate and extracted with DCM. The combined organic fractions were dried over Na2SO4 and The residue was purified by column chromatography on silica gel (DCM / MeOH 99:1 to 30:70) to give 102. LC / MS: (2M+H) + =613.56.

[0297] Step D - Synthesis of Intermediate 103 102 (220 mg, 0.718 mmol) and amber in DMF (4 ml) at room temperature A solution of 114 mg of acetic acid monomethyl ester (0.862 mmol) was added to HATU (300 mg, 0.790 mmol) and DIPEA (0.314 ml, 1.795 mmol). The mixture was stirred for 30 minutes and then diluted with saturated aqueous sodium bicarbonate at room temperature. The mixture was quenched with ethyl acetate and extracted with EtOAc. The combined organic fractions were washed with brine and The mixture was dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel. The product was purified by chromatography (eluting with hexanes / EtOAc 99:1 to 30:70). The reaction mixture was purified by filtration to give the intermediate, which was treated with 20% TFA in DCM for 1 h. was concentrated, then treated with 4N HCl (1.5 ml) and concentrated to give 103. LC / MS: (M+H) + =321.29.

[0298] Preparation of intermediate 109 [ka]

[0299] Step A - Synthesis of Intermediate 104 Methyl (S)-2-methylpyrrolidine-2-carboxylate in DMF (100 ml) at 0°C Silane hydrochloride (7.00 g, 39 mmol) and (S)-2-((tert-butyl) (Dicarbonyl)amino)-3-(4-methoxyphenyl)propanoic acid (12.08 g, 4 To a stirred solution of 0.9 mmol of ethyl acetate, DIPEA (17.01 ml, 97.0 mmol) was added. Then, HATU (19.26 g, 50.7 mmol) was added. The reaction was allowed to warm to room temperature and stirred overnight. The reaction was quenched with 10% aqueous LiCl. The organic extract was washed with 10% aqueous LiCl and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel (He Purification by hexanes / EtOAc (elution: 80:20 to 40:60) afforded 1 04 was provided.

[0300] Step B - Synthesis of Intermediate 105 A solution of 104 (16.4 g, 39.0 mmol) in EtOAc (100 mL) was added 4N HCl in oxane (48.8 ml, 195 mmol) was added. The resulting mixture The mixture was stirred at room temperature for 18 h and concentrated under reduced pressure to give 105, which was carried on to the next step. This was used without further purification.

[0301] Step C - Synthesis of Intermediate 106 105 (13.2 g, 37.0 mmol) and N-((benzyloxy)-2-methyl-2-propanediol) in DMF at 0 °C (oxy)carbonyl)-O-(tert-butyl)-L-threonine (18.15 g, 37 To a solution of 1.0 mmol of 1, DIPEA (16.15 ml, 92 mmol) was added, followed by HATU (18.28 g, 48.1 mmol) was added. The resulting mixture was left to stand. The reaction was then quenched with 10% aqueous LiCl and EtOAc. The organic extract was washed with 10% aqueous LiCl and dried over MgSO4. The solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica gel (hexanes / Purification by EtOAc (elution: 80:20 to 40:60) gave 106. did.

[0302] Step D - Synthesis of Intermediate 107 A solution of 106 (16.5 g, 27.0 mmol) in MeOH was added to a 10% Pd / C strip. Larry was added and the mixture was hydrogenated at 20 psi for 4 hours. The reaction mixture was filtered over Celite. The crude product was then redissolved in DCM and the solution was filtered through a 2 μm filter. Filtration through a filter and concentration gave 107.

[0303] Step E - Synthesis of Intermediate 108 A solution of 107 (3.2 g, 6.70 mmol) in DCM was treated with DIPEA (1.52 m l, 8.71 mmol) was added, followed by di-tert-butyl dicarbonate (1.90 g, 8.71 mmol) was added. The resulting mixture was stirred at room temperature for 2 hours and then cooled under reduced pressure. The residue was purified by column chromatography on silica gel (hexanes / EtOAc Purification by HPLC (elution: 100:0 to 40:60) gave 108.

[0304] Step F - Synthesis of Intermediate 109 108 (1.43 g, 2.475 ml) in THF (15 ml) and MeOH (15 ml) A solution of 40 mmol) and 1N LiOH (9.90 ml, 9.90 mmol) was Warmed to 5° C. and stirred for 4 h, then stirred at 32° C. for 48 h. The reaction was concentrated and cooled to 0° C. The mixture was quenched with 0.5M aqueous hydrochloric acid at rt until the pH reached approximately 2-3, and extracted with EtOAc. The combined organic fractions were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (hexanes / EtOAc-EtOH 99:1 to EtOAc-EtOH 3:1) to obtain 109. LC / MS: (M+H) + =564.49.

[0305] Preparation of intermediate compounds 110 to 115 Step A - Synthesis of Intermediate 110 [ka]

[0306] 109 (217 mg, 0.385 mmol) in DMF (2.5 ml), HATU ( 33 mg, 0.350 mmol) and DIPEA (0.245 ml, 1.400 mmol) A solution of 103 (217 mg, 0.385 mmol) was treated at 0° C., and the mixture was The mixture was allowed to warm to room temperature and stirred for 30 min. The mixture was quenched with aqueous ammonia and extracted with EtOAc. The residue was purified by precipitation with silica gel, washed with ethyl acetate, dried over Na2SO4, filtered and concentrated in vacuo. Column chromatography (hexanes / EtOAc-EtOH 3:1 99:1 Purification by elution with EtOAc-EtOH 3:1 from to give 110. LC / MS: (M+H) + =866.21.

[0307] Steps B and C - Synthesis of intermediate compounds 111 and 112 [ka]

[0308] To a solution of 110 (249 mg, 0.288 mmol) in DCM (1 ml) at room temperature was added HCl 4N in oxane (0.359 ml, 1.438 mmol) was added and the mixture was cooled to 6 Stirring for 1 h then concentrating gave 111. LC / MS: (M+H) + =7 10.19.

[0309] 111 (219 mg, 0.29 mL) in DMF (3 mL) and water (0.15 mL) at 0 °C A solution of 76 (232 mg, 0.285 mmol) and 77 (232 mg, 0.285 mmol) was added to DIPEA (0 .128 ml, 0.734 mmol) and HATU (123 mg, 0.323 mmol) ) was added and the mixture was stirred for 30 min. The mixture was quenched with brine at 0° C. and diluted with Et The combined organic fractions were dried over Na2SO4, filtered and concentrated in vacuo The residue was purified by column chromatography on silica gel (hexanes / EtOAc -EtOH 3-1 99:1 to 30:70, then DCM / MeOH 99:1 to Purification by HPLC (eluted at 70:30 from HPLC) gave 112. LC / MS: (M+H ) + =1506.11.

[0310] Step D - Synthesis of intermediate 113 [ka]

[0311] 112 in DCM (180 ml) and AcOH (15 ml) degassed with nitrogen for 30 min (173 mg, 0.115 mmol) was added to a solution of Zhan's catalyst (59.0 mg, 0.08 0 mmol) was added and the mixture was warmed to 50° C. and stirred for 3 h. The mixture was then cooled on Celite. It was filtered, washed with DCM and then concentrated in vacuo. The residue was purified by column chromatography on silica gel. Purification was performed by chromatography (eluting with DCM / MeOH 99:1 to 80:20). This gave 113 as a mixture of E and Z isomers. Isomers):(M) + = 1477.80; LC / MS (minor isomer): (M) + =1 478.28.

[0312] Step E - Synthesis of Intermediate 114 [ka]

[0313] To a solution of 113 (141 mg, 0.095 mmol) in acetonitrile (2 ml), Piperidine (0.066 ml, 0.668 mmol) was added and the mixture was stirred for 45 min. The mixture was concentrated in vacuo and coevaporated with acetonitrile tris(trice). This crude product was dissolved in DMF (2 ml) and water (0.1 ml) at 0° C. to give the crude product. (119 mg, 0.095 mmol) and intermediate compound 88 (52.0 mg, 0.1 05 mmol) was added to a slurry of HATU (39.7 mg, 0.105 mmol) and D IPEA (0.037 ml, 0.209 mmol) was added and the mixture was stirred for 30 min. The mixture was purified by column chromatography on C18 (acetonitrile + 0.05% TFA / water +0.05% TFA 90:10 to 30:70) to obtain 11 4 was given as a mixture of E and Z isomers. LC / MS major isomers: ( M) + = 1735.28; LC / MS (minor isomer): (M) + =1735.25 .

[0314] Step F - Synthesis of intermediate compound 115 [ka]

[0315] 114 (134 mg) in THF (1.5 ml) and MeOH (1.5 ml) at 0 °C 0.077 mmol) was added to a solution of 1N LiOH (0.386 ml, 0.386 m mol) was added dropwise and the mixture was stirred for 2 h. The reaction was cooled to 0 °C and diluted with 0.5 N HCl. Treat dropwise until the pH is about 7, concentrate from the organic solvent, and then add the slurry to about 1 mL Dissolved in DMF and purified by column chromatography on C18 (acetonitrile + 0.05% TFA / water + 0.05% TFA 90:10 to 50:50) LC / MS measurement gave 115 as a mixture of E and Z isomers. Positive isomer: (M) + = 1498.71; LC / MS (minor isomer): (M) + = 1499.48.

[0316] The R in each starting compound 2 Ex by reaction of amides with acidic substituent precursors The preparation of Ex-50 from Ex-01 and the preparation of Ex-55 from Ex-54 were carried out as described above. The following intermediate compounds can be used in similar reactions to further illustrate the utility of the present invention: Compounds for use may be provided.

[0317] R 1 / R 2 Synthesis of Substituent Precursors: 5-Carboxy-N-(3-methoxypropyl)-N,N-dimethylpentan-1-amine Preparation of nium chloride (intermediate Z-1a) Step A: Preparation of intermediate Z-1 [ka]

[0318] tert-Butyl 6-(dimethylamino)hexanoate in acetonitrile (1 mL) To a stirred solution of 1-bromo-3-methoxypropionate (300 mg, 1.393 mmol) was added Pan (853 mg, 5.57 mmol) was added. The reaction mixture was stirred at 50° C. for 16 h. The resulting mixture was concentrated under reduced pressure to give Z-1. LC / MS: (MB r)+=288.4. 1 H NMR(300MHz,CDCl3):δ3.76-3. 47(m,6H),3.38(d,J=28.9Hz,9H),2.25(t,J=7. 2Hz,2H),2.12-1.95(m,2H),1.87-1.55(m,4H), 1.45(s,11H).

[0319] Step B: Synthesis of intermediate compound Z-1a [ka]

[0320] To a stirred solution of Z-1 (460 mg, 1.249 mmol) in DCM (0.5 mL), 4M HCl in dioxane (2 mL) was added at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The residue was redissolved in DCM (5 mL) and concentrated under reduced pressure to give This gave intermediate compound Z-1a. LC / MS: (M-Cl) + =232.3.

[0321] Preparation of intermediate Z-2b Step A: Preparation of intermediate Z-2 [ka]

[0322] tert-Butyl 6-bromohexanoate (1.0 g, 3 To a stirred solution of 1.5 mL of dimethylamine (2M in THF) (7.96 mL, 15.0 mmol) was added The reaction mixture was stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 1%-1 Purification was performed by eluting with a gradient of 5% MeOH. Fractions containing the desired product were combined and concentrated to give Z-2. LC / MS: (M+H)+=216.2. 1 H NMR(300MHz,CDCl3):δ2.35-2.17(m,J=8.5 ,6.5Hz,10H),1.67-1.47(m,4H),1.45(s,9H),1 .42-1.23(m,2H).

[0323] Step B: Preparation of intermediate Z-2a [ka]

[0324] To a stirred solution of Z-2 (250 mg, 1.161 mmol) in ACN (1 mL), 1- Bromo-2-methoxyethane (645 mg, 4.64 mmol) was added. The reaction mixture was The mixture was stirred at 50° C. for 16 h. The resulting mixture was concentrated under reduced pressure to give Z-2a. LC / MS: (M-Br)+=274.3. 1 H NMR (300MHz, CDC l3):δ4.02-3.80(m,4H),3.70-3.54(m,2H),3.4 2(d,J=13.0Hz,9H),2.24(t,J=7.2Hz,2H),1.85 -1.75(m,2H),1.72-1.55(m,2H),1.44(s,11H).

[0325] Step C: Preparation of intermediate Z-2b [ka]

[0326] To a stirred solution of Z-2a (450 mg, 1.270 mmol) in DCM (0.5 mL) Then, 4M HCl in dioxane (2 mL) was added at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The residue was redissolved in DCM (5 mL) and concentrated under reduced pressure to give 3H 2 O. , giving Z-2b. LC / MS: (M-Cl) + =218.3.

[0327] Preparation of intermediate Z-3b Step A: Preparation of intermediate Z-3 [ka]

[0328] tert-Butyl 3-(2-hydroxyethoxy)propanoate in DCM (2 mL) A solution of CBr4 (1395 mg, 4.2 mmol) was added to a solution of 1,2-dichloro- ... 1 mmol) and PPh3 (965 mg, 3.68 mmol) were added at 0 °C. The mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography. The product was purified by chromatography using a gradient of 1%-15% EA in PE. The fractions containing the desired product were combined and concentrated to give Z-3. 1 H NMR(400MHz,CDCl3):δ3.78(dt,J=11.1,6.3 Hz,4H),3.47(t,J=6.3Hz,2H),2.53(t,J=6.4Hz ,2H),1.48(s,9H).

[0329] Step B: Synthesis of intermediate Z-3a [ka]

[0330] tert-Butyl 3-(2-bromoethoxy)propanoate in ACN (2 mL) To a stirred solution of Z-3 (450 mg, 1.778 mmol), trimethylamine (955 mM g, 5.33 mmol) (33% Wt in EtOH) was added. The reaction mixture was stirred at 50° C. for 1 The mixture was stirred for 6 h. The resulting mixture was concentrated under reduced pressure to give Z-3a. LC / MS:(M-Br)+=232.3. 1 H NMR (400MHz, CDCl3): δ 5.32(s,1H),4.04-3.94(m,4H),3.73(t,J=5.7H z,2H),3.50(s,10H),2.50(t,J=5.7Hz,2H),1.4 4(s,9H).

[0331] Step C: Synthesis of intermediate Z-3b [ka]

[0332] To a solution of Z-3a (550 mg, 1.761 mmol) in DCM (0.6 mL), 4M HCl in oxane (2.5 mL) was added at room temperature. The mixture was stirred at room temperature for 4 hours. The resulting mixture was concentrated under reduced pressure and the residue was dissolved in DCM (3 mL) and toluene (3 mL). The mixture was then concentrated under reduced pressure to give Z-3b. LC / M S: (M-Cl) + =176.2.

[0333] Preparation of intermediate Z-4b Step A: Preparation of intermediate Z-4 [ka]

[0334] To a solution of DIAD (1.755 mL, 9.03 mmol) in THF (30 mL), h3P (2.368 g, 9.03 mmol) was added. The mixture was stirred at room temperature for 10 min. Then methyl 2-(3-hydroxyphenyl)acetate (1.0 g, 6.02 mmol) l) and 3-(dimethylamino)propan-1-ol (0.931 g, 9.03 mm ol) was added to the solution. The mixture was stirred at 50° C. for 1 h. The resulting solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using 1%-10% MeO The product was purified by elution with a gradient of H. Fractions containing the desired product were combined and Concentration gave Z-4. LC / MS: (M+H)+=252.2. 1 HN MR(300MHz,CDCl3):δ7.23-7.18(m,1H),6.82(t d,J=8.7,4.1Hz,3H),4.01(t,J=6.4Hz,2H),3.6 9(s,3H),3.59(s,2H),2.46(t,J=7.3Hz,2H),2. 27(s,6H),1.97(dt,J=7.9,6.5Hz,2H).

[0335] Step B: Preparation of intermediate Z-4a [ka]

[0336] To a solution of Z-4 (600 mg, 2.268 mmol) in ACN (12 mL) was added MeI (1.288 g, 9.07 mmol) was added. The mixture was stirred at room temperature for 1 hour. The resulting solution was concentrated under reduced pressure to give -Z-4a. LC / MS: (MI) + =2 66.2. Step C: Preparation of intermediate Z-4b [ka]

[0337] To a solution of Z-4a (800 mg, 1.729 mmol) in THF (12 mL), 2M LiOH (1.729 mL, 3.46 mmol) was added, and the mixture was stirred at room temperature for 2 h. The pH value of the solution was adjusted to 4 with HCl (1M) and the solution was concentrated under reduced pressure. The product was purified by reversed-phase chromatography on C18 (mobile phase A: water, mobile phase B: ACN; flow rate: 6 0 mL / min; Gradient 1% B to 25% B in 25 min; 25% B to 95% B 15 min; 95% B to 95% B in 10 min) to give Z-4b. LC / MS: (M-Cl)+=252.2. 1 H NMR (300MHz, CD 3OD):δ7.21(t,J=7.9Hz,1H),6.95-6.75(m,3H) ,4.12(t,J=5.7Hz,2H),3.63-3.50(m,4H),3.18 (s,9H),2.35-2.20(m,2H).

[0338] Example 4 Preparation of Ex-23 [ka]

[0339] Step A: Preparation of intermediate S-1b (2S,3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H- Pyran-2,3,4,5-tetrayltetraacetate S-1a (5g, 12.81mm ol) in 48% HBr (7.25 mL, 64. The mixture was stirred at 0° C. for 1 hour. The reaction mixture was cooled in ice. Saturated aqueous sodium bicarbonate was added and the mixture was extracted with DCM (3 x 60 mL). The organic layer was washed with brine, dried over Na2SO4, filtered, concentrated, and then the crude product was The product was subjected to flash chromatography on silica gel (elution with 0-30% EtOAc / PE). The product was purified to S-1b by elution.

[0340] Step B: Preparation of intermediate S-1c To a solution of S-1b (4.5 g, 10.94 mmol) in anhydrous DMF (45 mL), Sodium disodium chloride (0.854 g, 13.13 mmol) was added and the reaction was incubated at 18° C. for 30 min. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3×80 mL). The organic layer was dried over Na2SO4 and evaporated to dryness. The crude product was purified by column chromatography on silica gel. Purify by rush chromatography (eluting with 0-30% EtOAc / PE) As a result, the drug was given a rating of S-1c.

[0341] Step C: Preparation of intermediate S-1d To a solution of S-1c (3.15 g, 8.44 mmol) in EtOH (60 mL), 10 %Pd-C (0.898 g, 0.844 mmol) was added. The reaction vessel was purged with air. The reaction was stirred at 18 °C for 5 h. The reaction mixture was cooled to 18 °C and charged with H2 at 50 psi. Dilution with tOAc, filtration through Celite, and concentration gave S-1d, which was Used for the next step.

[0342] Step D: Preparation of intermediate S-1e To a solution of S-1d (2.34 g, 6.74 mmol) in anhydrous THF (20 mL), Hydrofuran-2,5-dione (0.742 g, 7.41 mmol) and EtN(0 The reaction was stirred until the starting material was completely consumed. The mixture was stirred for 3 hours and then evaporated. The crude product was purified by flash chromatography on silica gel. S-1 was purified by chromatography (eluted with 0–10% DCM / MeOH) to give S-1 e. MS(ESI): m / z(M+H)+ 448.1. 1 HNMR(400 MHz, CDCl3) δ:6.48(d,J=9.04Hz,1H),5.43(s, 1H),5.24(t,J=8.93Hz,1H),5.07-5.17(m,2H), 4.08-4.18(m,2H),4.04(q,J=6.69Hz,1H),2.72 -2.84(m,1H),2.58-2.69(m,2H),2.43-2.53(m, 2H),2.15(s,3H),2.06(s,3H),2.04(s,4H),2.0 0(s,3H).

[0343] Step E: Preparation of Ex-23 Ex-01 (300 mg, 0.215 m A solution of S-1e (115 mg, 0.258 mmol) and S-1e was added with DIEA (0. (150 ml, 0.860 mmol) and HATU (98 mg, 0.258 mmol). The resulting solution was stirred at room temperature for 1 h. The reaction was diluted with 1N LiOH (2.58 ml, The mixture was quenched by dropwise addition of 2.58 mmol) and the resulting solution was stirred at room temperature for 2 h, then The filtrate was filtered and purified using a reverse phase HPLC C18 column with 29-34% TFA in water (0.05% TFA). Purification using a gradient of acetonitrile (0.05% TFA) afforded Ex-2 3 was obtained. LC / MS: [M+1]+=1657.1.

[0344] Example 5 Preparation of Ex-14 [ka]

[0345] Compound Ex-14 was prepared in a manner similar to that described in Example 1, but with different The "linkers" were prepared using alternative synthetic steps. The alternative steps and main assemblies are described below.

[0346] Preparation of intermediate Int-2g Intermediate Int-2g, useful as a "linker" in the preparation of compounds of the invention, is shown below: It was prepared according to the scheme: [ka]

[0347] Step A - Synthesis of Int-2gb Int-2da (0.5 g, 2 mmol) in THF (16 ml) at room temperature in a water bath and 2-azidoethanamine, a solution of HCl (0.246 g, 2 mmol) was added to the tri Sodium acetoxyhydroborate (1.06 g, 5 mmol) was added in portions and the mixture was The reaction was stirred for 2 h. The reaction was slowly quenched with saturated aqueous NaHCO3 and then with DCM. The combined organic layers were dried over MgSO4 and concentrated to give , giving Int-2gb. LC / MS: (M+1) + =320.3.

[0348] Step B - Synthesis of Int-2gc Int-2gb (0.64 g, 2 mmol) in DMF (4 ml) and DCM (8 ml) A solution of HATU (0. 914g, 2.4mmol) and DIPEA (0.7ml, 4.01mmol) The resulting solution was stirred at -15°C for 2 h, then quenched with water and concentrated. The residue was purified by reversed phase chromatography on C18 (acetonitrile / water + 0.1% TFA). Purification by elution with hexane gave Int-2gc. LC / MS: (M+1) + = 434.3.

[0349] Step C - Synthesis of Int-2g To a solution of Int-2gc (0.52 g, 1.2 mmol) in DCM (9 mL) at room temperature TFA (3 mL, 38.9 mmol) was added at rt. The reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure to give Int-2g. LC / MS: (M+1)+ = 334.3.

[0350] Preparation of intermediate compounds 70B and 76C [ka]

[0351] Step A - Synthesis of Intermediate 70B To a solution of 69B (1.5 g, 4.46 mmol) in DMF (17.8 ml) at 0° C. 95% NaH (0.141 g, 5.56 mmol) was added, and the resulting solution was incubated at 0° C. for 20 min. This was followed by stirring for 10 min, followed by 3-bromoprop-1-yne (80% in toluene) (0.596 ml, The resulting solution was added dropwise with an aqueous solution of lithium hydroxide (2M). (3345 μl, 6.69 mmol) was added dropwise. The reaction was stirred at room temperature for 2 hours. Filtration and purification by reverse phase HPLC (eluted with acetonitrile / water + 0.1% TFA) This gave 70B. LC / MS: (M+1) + : 361.0, (M+Na) + : 383.0.

[0352] Step B - Synthesis of Intermediate 76C The conversion of 70B to intermediate 76C is described in steps C to H of the preparation of intermediate 76. The procedure was similar to that described above. LC / MS: [M+1]+=812.16.

[0353] Assembly to Example Ex-14: [ka] TIFF2025041712000156.tif179169

[0354] Step A - Synthesis of Intermediate 117 Intermediate 117 was prepared from intermediates Int-2g and 76C as described in Examples 1 and 1A. Int-2g was prepared according to a procedure similar to that described previously. More specifically, Int-2g was prepared from intermediate 77B Further functionalization according to the reagents and procedures for the preparation of steps A to B yielded intermediate 86. Elaborate according to the procedure for preparation of steps G to J, then finally and intermediate 76C, which was coupled to intermediate 76D according to the procedure for the preparation of steps B to C of Example 1A. Catalysis gave 117. LC / MS: (M+1)+: 1573.36.

[0355] Step B - Synthesis of Intermediate 118 Copper tetrakis(acetonitrile) in BuOH (185.00 mL) / water (93 mL) (I) Hexafluorophosphate (51.6 mg, 0.138 mmol), Tris[( 1-Benzyl-1h-1,2,3-triazol-4-yl)methyl]amine (73.4 mg, 0.138 mmol) and sodium ascorbate (137 mg, 0.692 The mixture (435.3 mmol) was bubbled with nitrogen and then heated at 50° C. mg, 0.277 mmol) was added as a solid to the reaction. After 1 h, the reaction was pH The mixture was treated with aqueous buffer solution of 4 and extracted with EtOAc. The combined organic layers were evaporated to give a residue. eluted with a gradient of acetonitrile / water + 0.1% formic acid. to provide intermediate 118. LC / MS: (M+1)+: 157 3.2.

[0356] Step C-Synthesis of Ex-14 The synthesis of the example intermediate Ex-14 was carried out from intermediate 118 in a similar manner as described in Example 1. A similar procedure was followed to include the use of alternative spacers for assembly. C / MS:[M+1]+=1621.01.

[0357] Using the above synthetic scheme and the preparation of which may be described above, Some of the intermediates may be prepared with appropriate substitutions, including the use of alternative spacers that will be apparent to the art. As will be appreciated in some examples, the following compounds of the invention listed in Table 2 below were prepared: Additionally, alternative salt forms of the compounds of the present invention may also be described within this application.

[0358] Table 2 [Table 2] TIFF2025041712000158.tif170165TIFF2025041712000159.tif162166TIFF2025041 712000160.tif235166TIFF2025041712000161.tif224165TIFF2025041712000162.t if182165TIFF2025041712000163.tif221165TIFF2025041712000164.tif227164TIF F2025041712000165.tif153165TIFF2025041712000166.tif169165TIFF20250417120 00167.tif225165TIFF2025041712000168.tif222166TIFF2025041712000169.tif20 7165TIFF2025041712000170.tif229165TIFF2025041712000171.tif241170TIFF202 5041712000172.tif219165TIFF2025041712000173.tif226165TIFF20250417120001 74.tif221165TIFF2025041712000175.tif210166TIFF2025041712000176.tif146165

[0359] Determination of activity Selected compounds of the invention are assayed for their activity in antagonizing PCSK9 activity. To do so, the subject was subjected to one or more of the following procedures.

[0360] The following is a summary of the PCSK9 antagonism of the compounds of the present invention and any reported comparative compounds: Description of the assay used to determine activity of biotinylated PCSK9. I got what was on sale.

[0361] LDLR TR-FRET The PCSK9 TR-FRET assay measures the interaction between PCSK9 and LDLR. 40nM biotinylated PCSK9 + 10nM Lance ULight Str The solution containing eptavidin was diluted with 50 mM HEPES pH 7.4, 0.15M NaCl, 5 mM CaCl2, 0.01% BSA and 0.01% Surfac Prepare in tant P20. 40nM rhLDLR-6xHis+10nM Eu A separate solution containing -W1024 anti-6xHis was prepared in the same buffer system. Using the Echo, 0.750ul of compound is transferred to the assay plate followed by 1 Add 5ul of PCSK9+Ulight and 15ul of LDLR+Eu. The assay volume was 30.750 ul, with 20 nM PCSK9, 5 nM Ulight, Contains 20 nM LDLR and 5 nM Eu. Reactions are incubated at room temperature for at least 2 h. After incubation, the Envision Multilabel Reader was used Fluorescence measurements were performed. IC50 values ​​were determined by fitting the data to a sigmoidal dose-response curve using nonlinear regression. The europium-labeled LDLR is determined by fitting the The B counts are tracked to see if the compound has a deleterious effect on the LDLR. A decrease in the level of inhibition is likely a false positive indication.

[0362] Alexa FRET Standard TR-FRET The PCSK9 Alexa FRET Standard assay detects PCSK9 and Al Reagent A, an exaFluor647 (AF)-tagged cyclic peptide (K D =83nM) Measure the interaction between 1nM biotinylated PCSK9 and 2.5nM Lance A solution containing Streptavidin Europium (Strep-Eu) , 50mM HEPES pH7.4, 0.15M NaCl, 5mM CaCl2, 0 Made in 0.01% BSA and 0.01% Surfactant P20. Another solution containing 10 nM AlexaFluor-tagged cyclic peptide was added in the same buffer. Using Echo, transfer 0.750ul of compound to the assay plate. Then, 15ul of PCSK9+Stept-Eu and 15ul of AF peptide were added. The final assay volume was 30.750 ul, containing 0.5 nM PCSK9, 1.2 The reaction contained 5 nM Strep-Eu and 20 nM AF cyclic peptide. After incubation for at least 2 hours in Envision Multilabel Fluorescence measurements are performed using a reader. IC50 values ​​are calculated using nonlinear regression for the sigmoidal Ki is then determined by fitting the data to a dose-response curve. IC50 and K of F cyclic peptides D Europium-labeled PCSK Track the B count to see if the compound has a negative effect on PCSK9 A decrease in B counts is likely to indicate a false positive for inhibition. Data from this procedure is reported as A = 'number' (nanomolar concentration).

[0363] Reagent A was prepared according to the following method: [ka]

[0364] Step A - Synthesis of intermediate compound Int-A Peptides were analyzed on a 0.250 mmol scale using a CEM Liberty Blue microscope. Fmoc / tBu was synthesized on PS Rink-Amide MBHA resin on a microwave synthesizer. Using chemistry, 0.32 mmol g -1 The assembly was carried out using 4 equiv. 0.2M Fmoc-protected amino acid in DMF, 4 eq. 0.5M HATU in DMF, 4 eq. The reaction was carried out using a single coupling with 2M DIPEA (for Tyr, The Fmoc deprotection cycle consisted of 20% (V / V) piperidine in DMF. The experiment was carried out using gin.

[0365] The Fmoc-protected amino acids and building block sequences used are as follows: 1. N-(((9H-fluoren-9-yl)methoxy)carbonyl)-S-trityl -L-Cysteine 2. (S)-1((((9H-fluoren-9-yl)methoxy)carbonyl)amino )-2-Methylpyrrolidine-2-carboxylic acid 3. (((9H-Fluoren-9-yl)methoxy)carbonyl)-L-tyrosine 4. N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-trityl -L-histidine 5. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amine (o)-4-(tert-butoxy)-4-oxobutanoic acid 6. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amine 5-Fluoro-1H-indol-3-ylpropanoic acid 7. (S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amine 5-Fluoro-1H-indol-3-ylpropanoic acid 8. (((9H-Fluoren-9-yl)methoxy)carbonyl)glycine 9. N 2 -(((9H-fluoren-9-yl)methoxy)carbonyl)-N 6 -(t (ert-Butoxycarbonyl)-L-lysine 10. 3-(Tritylthio)propanoic acid

[0366] At the end of the assembly, the resin was washed with DMF, MeOH, DCM, Et2O. The tides were dissolved in 50 ml of TFA solution (v / v) (91% TFA, 5% HO, 4% TIPS). The resin was cleaved from the solid support using 1.5% CO at room temperature for approximately 1.5 hours. The resin was filtered and washed with TFA. The solution was concentrated to dryness and freeze-dried. g), which was used crudely in the next step. LCMS analysis C61H Calculated value for 75F2N15O13S2: 1328.48, Measured value: 1328.2 (M+1) +

[0367] Step B - Synthesis of intermediate compound Int-B: As described for Reagent B RP-HPLC (Waters Deltapak C4, double cartridge, 40 x 100mm, 15m, 300A; 15% to 35% ACN / water + 0.1% over 20 minutes The collected fraction was lyophilized to give 35 mg of 100% 1,000 mg of ... To give intermediate compound Int-B. LCMS analysis of C69H81F2N15O13S2 Calculated: 1430.62; Measured: 1430.9 (M+1) +

[0368] Step C - Synthesis of compound Reagent A: As described for Reagent B LCMS analysis C105H122F2N17O26S6 3- Calculated value: 2268.58 ;1135.8(M+2) 2+

[0369] Alexa FRET Plus TR-FRET The PCSK9 Alexa FRET Plus assay detects PCSK9 and AlexaF Reagent B, a luor647 (AF)-tagged cyclic peptide (K D = 35nM) Measure the interaction: 1nM biotinylated PCSK9 + 2.5nM Lance Str A solution containing eptavidin Europium (Strep-Eu) was added to 50 ml M HEPES pH7.4, 0.15M NaCl, 5mM CaCl2, 0.01% Made in BSA and 0.01% Surfactant P20. A separate solution containing AlexaFluor-tagged cyclic peptides was added in the same buffer system. Prepare the mixture in a flask using Echo. Transfer 15ul of PCSK9+Stept-Eu and and 15 ul of AF peptide. The final assay volume is 30.750 ul. 0.5nM PCSK9, 1.25nM Strep-Eu and 960nM AF cyclic The reaction was incubated at room temperature for at least 2 hours, after which Envi Fluorescence measurement was performed using a sion Multilabel Reader. IC50 values ​​were by fitting the data to a sigmoidal dose-response curve using nonlinear regression. Ki is then determined by the IC50 and K D It is calculated from Europium-labeled PCSK9 counts (B counts) were tracked to determine whether the compound inhibited PCSK9. Observe whether the antibody has any adverse effect on SK9. A decrease in B counts may indicate a false positive for inhibition. Data from this procedure are reported as "P = 'number' (nanomolar concentration)." can be.

[0370] Reagent B was prepared by the following procedure. [ka]

[0371] Step A - Synthesis of intermediate compound Int-A Peptides were analyzed on a 0.250 mmol scale using a CEM Liberty Blue microscope. Fmoc / tBu was synthesized on PS Rink-Amide MBHA resin on a microwave synthesizer. Using chemistry, 0.32 mmol g -1 The assembly was carried out using 4 equiv. 0.2M Fmoc-protected amino acid in DMF, 4 equivalents of 1M oxime in DMF, 4 equivalents of 0. Single coupling using 5M N,N-diisopropylcarbodiimide (DIC) (double coupling for Y01). The Fmoc deprotection cycle was , was carried out using 20% ​​(V / V) piperidine in DMF.

[0372] The Fmoc-protected amino acids and building block sequences used are as follows: 1. N-(((9H-fluoren-9-yl)methoxy)carbonyl)-S-trityl- L-Cysteine 2. (S)-1((((9H-fluoren-9-yl)methoxy)carbonyl)amino) -2-Methylpyrrolidine-2-carboxylic acid 3. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino )-3-(4-Methoxyphenyl)propanoic acid 4. N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-trityl- L-Histidine 5. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino )-4-(tert-butoxy)-4-oxobutanoic acid 6. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino )-3-(5-fluoro-1H-indol-3-yl)propanoic acid 7. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino )-3-(5-fluoro-1H-indol-3-yl)propanoic acid 8. (((9H-Fluoren-9-yl)methoxy)carbonyl)-D-alanine 9.N 2 -(((9H-fluoren-9-yl)methoxy)carbonyl)-N 6 -(te rt-Butoxycarbonyl)-L-lysine 10. 3-(Tritylthio)propanoic acid

[0373] At the end of the assembly, the resin was washed with DMF, MeOH, DCM, and Et2O. The solution was diluted with 50 ml of TFA solution (v / v) (91% TFA, 5% H2O, 4% TIPS). The resin was cleaved from the solid support using 100% TFA at room temperature for approximately 1.5 hours. The resin was filtered and washed with TFA. The solution was concentrated to dryness and freeze-dried. ) which was used crudely in the next step. LCMS analysis C63H7 Calculated value for 9F2N15O13S2: 1356.53, Measured value: 1356.9 (M+1) +

[0374] Step B - Synthesis of intermediate compound Int-B Crude Int-A (0.22 mmol) was redissolved in 24 ml of DMF. 1 of 1M sodium bicarbonate solution was added to raise the pH to 7. of 1,3-bis(bromomethyl)benzene (0.1 M in DMF) was added dropwise. The mixture was stirred at room temperature for 20 min and quenched with TFA (until pH was 3-4). Concentration in vacuo then afforded crude Int-B, which was purified by RP-HPLC ( Waters XBridge, C18, 50×150mm, 5μm, 130A; 20 minutes The mixture was purified by elution with 25% to 40% ACN / water + 0.1% TFA (adjustment) over a period of 10 minutes. The fractions were lyophilized to give 35 mg of intermediate compound Int-B. CMS analysis: Calculated for C71H85F2N15O13S2: 1458.67; Found: 1 458.8(M+1)+

[0375] Step C - Synthesis of Reagent B The intermediate compound Int-B (15 mg) was dissolved in 0.2 ml of anhydrous DMSO. 15 mg of ALEXAFLUOR 647 dissolved in 1.5 ml of anhydrous DMSO NHS Ester (A37566, Life Technology) was added. 1 μL of anhydrous DIPEA was added. The reaction was stirred at room temperature for 12 hours under nitrogen atmosphere. The mixture was then quenched with TFA (until the pH reached 3-4) and purified by RP-HPLC (Dr Maish, Reprosil Gold C18, 250×20mm, 120Å, 1 0 μm; 20% to 35% 0.1% TFA in ACN / 0.1% T in H2O over 20 min FA, then 35% to 40% over 5 min, flow rate of 20 mL / min). The collected fraction was lyophilized to give 16.1 mg of compound B. MS analysis C107H126F2N17O26S6 3- Calculated value: 2296.64; measured Value: 1150.6(M+2) 2+

[0376] Activity data obtained by one or both of the above procedures may be used to assess the activity of selected The example compounds are reported in the following format: Example No.: A(standard TR Fret) = 'number'; P(Alexa Fr et plus standard TR Fret) = 'number' / all reported All values ​​are nanomolar.

[0377] Alexa FRET Ultra TR-FRET The PCSK9 Alexa FRET Ultra Assay detects PCSK9 and Alexa Reagent B, a Fluor647 (AF) tagged cyclic peptide (K D =0.99nM) Measure the interaction between 1nM biotinylated PCSK9 and 2.5nM Lance A solution containing Streptavidin Europium (Strep-Eu) 50mM HEPES pH7.4, 0.15M NaCl, 5mM CaCl2, 0. Made in 0.1% BSA and 0.01% Surfactant P20. Another solution containing 10 nM AlexaFluor-tagged cyclic peptide was added in the same buffer. The mixture is prepared in a 0.015ul system using Echo. SO was transferred to each well of the assay plate, followed by 15 ul of PCSK9+Stept- Add Eu and 15ul of AF peptide. The final assay volume is 30.750ul. Yes, 0.5nM PCSK9, 1.25nM Strep-Eu and 960nM A The reaction was incubated at room temperature for at least 2 hours, then incubated at 4°C for 1 h. Fluorescence measurements are performed using the nvision Multilabel Reader. IC5 The zero value was determined by fitting the data to a sigmoidal dose-response curve using nonlinear regression. Ki is then determined by the IC50 and K D Calculated from Europium-labeled PCSK9 counts (B counts) were tracked to determine the compound Observe whether the B counts are negatively affected by PCSK9. A decrease in B counts indicates a false positive for inhibition. Data from this procedure are likely to be Data are reported as "nanomolar concentrations."

[0378] The following compounds, shown in Table 2, were evaluated using the above protocol. The results are shown below. show: Ex-01 Ki Plus=<0.00558, Ki Ultra=0.0046 / E x-02 Ki Plus=0.00558、Ki Ultra=0.005933 / E x-03 Ki Plus=0.02535、Ki Ultra=0.06803 / Ex -04 Ki Plus ≦0.00558、Ki Ultra=0.004711 / E x-05 Ki Plus=0.009621、Ki Ultra=0.03296 / E x-06 Ki Plus=0.00568、Ki Ultra=0.003424 / E x-07 Ki Plus=0.05914、Ki Ultra=0.06753 / Ex -08 Ki Plus=0.01574、Ki Ultra=0.06832 / Ex- 09 Ki Plus=0.09189、Ki Ultra=0.247 / Ex-10 Ki Plus=0.005743、Ki Ultra=0.02489 / Ex-11 Ki Plus=0.04334、Ki Ultra=0.2067 / Ex-12 Ki Plus=0.01448、Ki Ultra=0.02247 / Ex-13Ki Plus=0.1454、Ki Ultra=0.4772 / Ex-14 Ki Plus s=0.01605、Ki Ultra=0.02099 / Ex-15 Ki Plus =0.1027、Ki Ultra=0.2601 / Ex-16 Ki Plus=0. 01423、Ki Ultra=0.05141 / Ex-17 Ki Plus≦0. 00558、Ki Ultra=0.0028 / Ex-18 Ki Plus=0.03 356、Ki Ultra=0.1183 / Ex-19 Ki Plus=0.0166 2、Ki Ultra=0.01204 / Ex-20 Ki Plus=0.01303 、Ki Ultra=0.01711 / Ex-21 Ki Plus=0.005692 、Ki Ultra=0.001264 / Ex-22 Ki Plus=0.00926 、Ki Ultra=0.01519 / Ex-23 Ki Plus=0.00938、 Ki Ultra=0.00239 / Ex-24 Ki Plus=0.00812、K and Ultra=0.00767 / Ex-25 Ki Plus=0.01127、Ki Ultra=0.00463 / Ex-26 Ki Plus ≦0.00558、Ki Ultra=0.002754 / Ex-27 Ki Plus ≦0.00558、K i Ultra=0.00301 / Ex-28 Ki Plus ≦0.00558、K and Ultra=0.00078 / Ex-29 Ki Plus=0.00981、Ki Ultra=0.00614 / Ex-31 Ki Plus <0.00558、Ki Ultra=0.00074 / Ex-35 Ki Plus=0.04652、Ki Ultra=0.08434 / Ex-36 Ki Plus=0.00762、Ki U ltra=0.00507 / Ex-38 Ki Plus=0.00904、Ki Ul tra=0.01416 / Ex-39 Ki Plus=0.00716、Ki Ult ra=0.00414 / Ex-40 Ki Plus=0.30800、Ki Ultr a=0.86010 / Ex-41 Ki Plus=0.00697、Ki Ultra =0.00628 / Ex-44 Ki Plus=0.01445、Ki Ultra= 0.02194 / Ex-47 Ki Plus=0.01474、Ki Ultra=0 .01193 / Ex-48 Ki Plus=0.01169、Ki Ultra=0. 01545 / Ex-49 Ki Plus=0.00716、Ki Ultra=0.0 0414 / Ex-50 Ki Standard <1.26, Ki Plus=0.0 1052、Ki Ultra=0.00443 / Ex-51Ki Standard <1.26、Ki Plus <0.00558、Ki Ultra=0.00597 / Ex-52 Ki Standard <1.26, Ki Plus <0.00558 、Ki Ultra=0.00359 / Ex-53 Ki Standard <1.2 6、Ki Plus=0.09629、Ki Ultra=0.21500 / Ex-54 Ki Standard<1.26、Ki Plus=0.36720、Ki Ul tra=0.48390 / Ex-55 Ki Standard <1.26、Ki P lus=0.07240、Ki Ultra=0.23800 / Ex-56 Ki St andard <1.257、Ki Plus=0.02237、Ki Ultra=0 .00481 / Ex-57 Ki Standard <1.257、Ki Plus <0.00558、Ki Ultra=0.00162 / Ex-58 Ki Stand ard <1.257、Ki Plus=0.00773、Ki Ultra=0.00 196 / Ex-59 Ki Plus=0.00788、Ki Ultra=0.004 959 / Ex-60 Ki Plus=0.006515、Ki Ultra=0.00 5312 / Ex-61 Ki Plus=0.00747、Ki Ultra=0.00 6543.

Claims

1. A combination comprising a compound of formula I and an additional active agent, wherein the additional active agent is an antihypertensive or antiatherosclerotic agent; and wherein the compound of formula I is a compound having the following structure or any pharma- ceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, X is H, F, Cl or Br; R １ is selected from the following: (a)-H; or (b) -(CH ２ ) ｚ -R １４Ａ where z is 1-6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ())? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ in be); (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ and (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (aii)-N ＋ (CH ３ ) ３ ; or (aiii) Formula: 【Chemistry 2】 (part of R ２ is selected from the following: (a)-H; and (b) -(CH ２ ) ｚ -R １４Ａ where z is 1 to 6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ())? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (Wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ and (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (ai)-N ＋ (CH ３ ) ２ R １４ｃａ (In the formula, R １４ｃａ is -CH ３ or - (CH ２ ) １－４ -OCH ３ ) (aiii) Formula: 【Chemistry 3】 part; or (aiv) Formula: 【Chemistry 4】 (wherein R １４Ｃｂ and R １４Ｃｃ is selected from 1 to 4); and is R １ and R ２ are bonded together to form the formula: 【Chemistry 5】 may form a portion of the formula: G １ , R Ｇ１ａ and R Ｇ１ｂ is defined as follows: (a) G １ is the formula: 【Chemistry 6】 wherein n ｑ１ is 1 to 6, m ｑ１ is 0, 1 or 2, and together they equal n ｑ１ and m ｑ１ are selected such that the length of the linker moiety they define does not exceed a total of eight carbon and / or oxygen atoms making up the chain, including the carbon atom in the chain forming the carbonyl moiety; R Ｇ１ａ is selected from: (i) —H; and (ii) alkyl of up to 4 carbon atoms; and R Ｇ１ｂ teeth: (i) Formula: 【Chemistry 7】 part; and (ii) Formula: 【Chemistry 8】 or (b) G １ is the formula: 【Chemistry 9】 is a linker moiety, ｑ２ is 0, 1 or 2; m ｑ２ are 1 to 6, and together they make up n ｑ２ and m ｑ２ are selected such that the length of the linker moiety they define does not exceed a total of eight carbon and / or oxygen atoms making up the chain, including the carbon atom in the chain forming the carbonyl moiety; R Ｇ１ａ teeth: (i) Formula: 【Chemistry 10】 part; and (ii) Formula: 【Chemistry 11】 and R Ｇ１ｂ is selected from: (i) —H; and (ii) alkyl of up to 4 carbon atoms; R ８ is -CH ３ Or the formula: 【Chemistry 12】 where R ８ａ is -H or alkyl of up to 4 carbon atoms, straight, branched or cyclic; A: (a) Formula: 【Chemistry 13】 part; (b) -CH ２ - (CH ２ ) ｙ -CH ２ - (wherein y is 1 to 6); (c) Formula: 【Chemistry 14】 (wherein A ｂ１ teeth: (i) Formula: 【Chemistry 15】 where x is 1 to 6; or (ii) Formula: 【Chemistry 16】 where y is 1 to 5; (d) Formula: -CH ２ - (CH ２ ) ｍ -O-(CH ２ ) ｎ - moiety (wherein m is 1 to 5 and n is 0 or 1 to 4). Selected from: B is: (a) combination; (b) -(CH ２ ) １－２ ;or (c) Formula: 【Chemistry 17】 is a portion of D is: (a) Formula: 【Chemistry 18】 (wherein E is -CH ２ - or - (CH ２ ) ２－４ -O-, where A and B are as defined above; (b) Formula: 【Chemistry 19】 wherein A and B are as defined above; (c) Formula: 【Chemistry 20】 (wherein, n ａ is 1, 2 or 3; m ａ is 2, 3 or 4, n ａ +m ａ is ≧3, where A and B are as defined above; or (d) Formula: 【Chemistry 21】 (wherein R ３４ｂ is -H or a straight, branched or cyclic alkyl of up to 4 carbon atoms, and A and B are as defined above. The combination.

2. 2. The combination of claim 1, wherein the compound of formula I is a compound having the structure of formula IIE or a pharma- ceutically acceptable salt thereof; 【Chemical 22】 [In the formula, R １ is selected from the following: (a)-H; or (b) -(CH ２ ) ｚ -R １４Ａ where z is 1-6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ())? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ in be); (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ and (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (ai)-N ＋ (CH ３ ) ３ ; and R ２ is selected from the following: (a)-H; and (b) -(CH ２ ) ｚ -R １４Ａ where z is 1 to 6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ()))? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (Wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ and (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (ai)-N ＋ (CH ３ ) ２ R １４ｃａ (In the formula, R １４ｃａ is -CH ３ or - (CH ２ ) １－４ -OCH ３ ) ; A is selected from the following: (a) -CH ２ - (CH ２ ) ｙ -CH ２ - (wherein y is 1 to 6); (b) Formula: 【Chemistry 23】 (wherein A ｂ１ teeth: (i) Formula: 【Chemistry 24】 where x is 1 to 6; or (ii) Formula: 【Chemistry 25】 where y is 1 to 5; (c) Formula: -CH ２ - (CH ２ ) ｍ -O-(CH ２ ) ｎ - moiety, wherein m is 1 to 5 and n is 0 or 1 to 4; The combination.

3. R １ teeth: (a) -(CH ２ ) ｚ -R １４Ａ where z is 1-6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ()))? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ in be); (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ and (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (ai)-N ＋ (CH ３ ) ３ ; and R ２ teeth: (a) -(CH ２ ) ｚ -R １４Ａ where z is 1 to 6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ()))? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (Wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ or (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (ai)-N ＋ (CH ３ ) ２ R １４ｃａ (In the formula, R １４ｃａ is -CH ３ or - (CH ２ ) １－４ -OCH ３ ) ; A: (a) -CH ２ - (CH ２ ) ｙ -CH ２ - (wherein y is 1 to 6); The combination according to claim 2,

4. R １ teeth: (a) -(CH ２ ) ｚ -R １４Ａ where z is 1-6 and R １４Ａ teeth: (i) -H; R ２ teeth: (a) -(CH ２ ) ｚ -R １４Ａ where z is 1 to 6 and R １４Ａ teeth: (i) -H; ((ii)-NH) ２ ; ())? ＋ H ３ 4 (iv)-N ＋ (H) ３ C) ３ ; (v)-NH-C(O)-[(CH ２ ) ２ -O-] ２ - (CH ２ ) ２ R １４Ｂ (In the formula, R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vi)-NH-C(O)-[(CH ２ ) ｙ１２ -O-] ２ - (CH ２ ) ｙ１３ R １４Ｂ (Wherein: y12 and y13 are not both simultaneously 2 and are independently 2 to 4; and R １４Ｂ Is: -NH ２ ;-N ＋ H ３ ; -N(CH ３ ) ２ ; or -N ＋ (CH ３ ) ３ ) (vii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ is -O-(CH ２ ) ３－４ -N ＋ (CH ３ ) ３ or (viii)-NH-C(O)-(CH ２ ) ｙ R １４Ｃ (wherein y is 1 to 6; R １４Ｃ teeth: ((ai)-O-(CH) ２ ) ２ -N ＋ (CH ３ ) ３ ; (ai)-N ＋ (CH ３ ) ２ R １４ｃａ (In the formula, R １４ｃａ is -CH ３ or - (CH ２ ) １－４ -OCH ３ ) ; A: (a) -CH ２ - (CH ２ ) ｙ -CH ２ - (wherein y is 1 to 6); The combination according to claim 3,

5. 2. The combination according to claim 1, wherein the compound of formula I is selected from: 【Chemistry 26】 【change】 【change】 Here, A － is a pharma-ceutically acceptable anion.

6. The combination of any of claims 1 to 5, wherein the additional active agent is a lipid lowering agent, a cholesterol absorption inhibitor, an HMG-CoA reductase inhibitor, an immediate release or controlled release form of niacin; a niacin receptor agonist or partial niacin receptor agonist, a PPARα agonist, or a bile acid sequestrant.

7. 7. The combination of any of claims 1 to 6, wherein the additional active agent is selected from simvastatin, lovastatin, atorvastatin, rosuvastatin, pravastatin, fluvastatin, cerivastatin and pitavastatin.

8. The combination of any of claims 1 to 7, wherein the additional active agent is rosuvastatin.

9. The combination of any of claims 1 to 8, wherein the compound of formula I and the additional active agent are in a single dosage formulation.

10. The combination of any of claims 1 to 8, wherein the compound of formula I and the additional active agent are in separate dosage formulations.

11. A combination comprising a compound of formula I having the following structure: 【Chemistry 27】 Here, A － is a pharma- ceutically acceptable anion; The combination comprises an additional active agent, wherein the additional active agent is an antihypertensive agent or an antiatherosclerotic agent. Combination agents.

12. The compound of formula I is: 【Chemistry 28】 The combination according to claim 11.

13. 13. The combination of claim 11 or 12, wherein the additional active agent is a lipid lowering agent, a cholesterol absorption inhibitor, an HMG-CoA reductase inhibitor, an immediate release or controlled release form of niacin, a niacin receptor agonist or partial niacin receptor agonist, a PPARα agonist, or a bile acid sequestrant.

14. 14. The combination of any of claims 11 to 13, wherein the additional active agent is selected from simvastatin, lovastatin, atorvastatin, rosuvastatin, pravastatin, fluvastatin, cerivastatin and pitavastatin.

15. The combination of any of claims 11 to 14, wherein the additional active agent is rosuvastatin.

16. The combination of any of claims 11 to 15, wherein the compound of formula I and the additional active agent are in a single dosage formulation.

17. The combination of any of claims 11 to 15, wherein the compound of formula I and the additional active agent are in separate dosage formulations.

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