Late-stage peptide cyclization for disulfide mimetic formation

Abstract

This invention discloses a method for constructing disulfide mimetic compounds for late cyclization of peptides. The disclosed method provides late cyclization via thioacetalization of peptides containing two cysteine ​​residues to form a thioacetal group. These bonds are stable under acidic, basic, and reducing conditions. A wide range of cyclic ketones, and even acetone, can be used as crosslinking agents in this reaction. Since trifluoroacetic acid (TFA) can function as both a catalyst and the sole solvent, most peptide sequences can be cyclized by the disclosed method. The disclosed method offers many advantages and merits, including the use of TFA as a robust catalyst and the sole required solvent, the readily available availability of acetone and a variety of cyclic ketones as reactants, good chemoselectivity and tolerance of natural peptides to this reaction, and the fact that the resulting crosslinks are structural mimetic compounds of disulfides.

Description

[Technical Field]

[0001] [Cross-reference of related applications] This application claims the benefit and priority of U.S. Provisional Application No. 63 / 467,583, filed on 18 May 2023, and all contents of said Provisional Application are incorporated herein by reference for all purposes.

[0002] The disclosed inventions generally relate to the field of protein and peptide crosslinking, and more particularly to the field of peptide crosslinking using late disulfide mimetic compounds. [Background technology]

[0003] In the late modification of peptides, cyclization is an important strategy for increasing the rigidity of the three-dimensional structure compared to linear peptides. The reduced flexibility allows the peptide to be fixed to its three-dimensional structure, improving stability and resistance to protease degradation. Peptide disulfide cyclization is a powerful technique in drug development, involving the formation of cyclic peptides through the formation of a disulfide bond between two cysteine ​​residues in the peptide chain. This process generates more stable and structurally constrained molecules that exhibit improved pharmacological properties, such as enhanced potency, selectivity, and stability. In recent years, peptide disulfide cyclization has attracted significant attention due to its potential in developing novel therapies for various diseases, including cancer, infectious diseases, and metabolic disorders. This approach has led to the successful development of several FDA-approved drugs, such as insulin, oxytocin, and vasopressin. However, the inherent instability of disulfides in reducing environments limits their broad applications and widespread distribution.

[0004] Any discussion of documents, acts, materials, apparatus, articles, etc. contained herein, which existed prior to the priority date of each claim of this disclosure, should not be construed as forming part of the basis of the prior art or being common knowledge in the art related to this disclosure.

[0005] Throughout this specification, the word “comprise,” or variations such as “comprises” or “comprising,” shall be understood to mean including the aforementioned elements, integers, steps, or groups of elements, integers, or steps, but not to exclude other elements, integers, steps, or groups of elements, integers, or steps. [Overview of the project]

[0006] This invention discloses a method for constructing disulfide mimetic compounds for late-stage cyclization of peptides. Natural disulfide bonds are unstable under reducing conditions, limiting their broad applications. The disclosed late-stage cyclization involves thioacetalization of a peptide containing two cysteine ​​residues, forming a thioacetal group stable under acidic, basic, and reducing conditions. Compared to existing methods, the disclosed peptide thioacetalization can generate novel disulfide mimetic compounds. A wide range of cyclic ketones, and even acetone, can be used as crosslinking agents in this reaction. Since trifluoroacetic acid (TFA) can function as both a catalyst and the sole solvent, virtually all peptide sequences can be used in the disclosed method.

[0007] The disclosed method offers numerous advantages and merits, including the use of TFA as a robust catalyst and the only solvent required for the reaction, the readily available availability of acetone and a variety of cyclic ketones as reactants, good chemoselectivity and tolerance of natural peptides to this reaction, and the fact that the resulting crosslinks are structural mimics of disulfides.

[0008] This invention discloses a method for the late cyclization of peptides and reagents used in the method. Generally, the method involves maintaining a reaction mixture at a temperature sufficient to form a product for a sufficient amount of time. Generally, the reaction mixture comprises a peptide containing two or more cysteine ​​residues, a cyclic ketone reagent or acetone, and a solvent. Generally, the product comprises a thioacetalized peptide, in which two cysteine ​​residues of the peptide are linked by thioacetalization.

[0009] In some forms, peptides are linear or cyclic (including monocyclic and bicyclic structures). In some forms, the peptide is a random peptide or a peptide drug. In some forms, the peptide is formed from natural amino acids.

[0010] In some forms, cyclic ketone reagents are cyclic ketone reagents having the following structure: JPEG2026521678000001.jpg3024 Formula I In the formula, R and R' are independently alkyl groups (e.g., C1-C6 alkyl groups), and TIFF2026521678000002.tif212 is absent, or R and R' together form a cyclic moiety A, where A is a cycloalkyl group, cycloalkenyl group, cycloalkynyl group, aryl group, polyaryl group, heteroaryl group, heteropolyaryl group, or heterocyclic group, where R'' represents hydrogen or a substituent on the cyclic moiety A, and R'' is, whenever it appears, independently a benzyl group, an allyl group, an alkyl group (e.g., a C1-C6 alkyl group), a halogen, -CN, -CF3, -NO2, an alkoxy group, or an aryl group (e.g., a phenyl group), and n is an integer between 0 and 10, 0 and 8, 0 and 6, 0 and 4, or 0 and 2. If R'' is an alkyl group, the alkyl group may be substituted or unsubstituted, and the substituent, if present, may be any substituent disclosed herein, such as a halogen, an azide group, or an alkynyl group (e.g., -CCH or -CH2CCH).

[0011] In some forms, R and R' are independently alkyl groups such as methyl groups. In some forms, R and R' are independently alkyl groups such as methyl groups, and TIFF2026521678000003.tif212 does not exist. In some forms, A is cyclobutane, azetidine, cyclopentane, fluorenone, cyclohexane, or piperidine.

[0012] In some forms, the solvent is trifluoroacetic acid. In some forms, trifluoroacetic acid acts as a catalyst and is the only solvent in the reaction mixture.

[0013] In some forms, the reaction mixture is maintained at a temperature of 20°C to 35°C, such as approximately 30°C, for a maximum of 1 hour, 2 hours, 3 hours, 10 minutes to 1 hour, 20 minutes to 2 hours, or 30 minutes to 3 hours.

[0014] In some forms, the molar ratio of peptide to cyclic ketone reagent (peptide:cyclic ketone reagent) is between 0.1 and 1, for example, about 0.2.

[0015] In some forms, cyclic ketone reagents have one of the following structures: JPEG2026521678000004.jpg32120 Here, R1 and R3 are H, a benzyl group, an allyl group, or an alkyl group (e.g., a C1-C6 alkyl group), and each R2 is independently H, a benzyl group, an allyl group, an alkyl group (e.g., a C1-C6 alkyl group), a halogen, -CN, -CF3, -NO2, an alkoxy group, or an aryl group. If any of R1-R3 is an alkyl group, the alkyl group may be substituted or unsubstituted, and the substituent, if present, may be any substituent disclosed herein, such as a halogen, an azide group, or an alkynyl group (e.g., -CCH or -CH2CCH).

[0016] In some forms, thioacetalized peptides have one of the following structures: JPEG2026521678000005.jpg5891、 JPEG2026521678000006.jpg6980、 JPEG2026521678000007.jpg6994、 JPEG2026521678000008.jpg6899、 JPEG2026521678000009.jpg5495、 JPEG2026521678000010.jpg6368、 JPEG2026521678000011.jpg6294、 JPEG2026521678000012.jpg5889、 JPEG2026521678000013.jpg5889、 JPEG2026521678000014.jpg5990、 JPEG2026521678000015.jpg6086、 JPEG2026521678000016.jpg6299、 JPEG2026521678000017.jpg6185、 JPEG2026521678000018.jpg6773、 JPEG2026521678000019.jpg6368、 JPEG2026521678000020.jpg69127、 or JPEG2026521678000021.jpg8885。

[0017] Regarding other advantages of the disclosed methods and compositions, some are described in the following description, some can be understood from the description, or can be acquired by implementing the disclosed methods and compositions. The advantages of the disclosed methods and compositions can be realized and obtained by the elements and combinations specifically pointed out in the appended claims. It should be understood that the foregoing general description and the following detailed description are merely exemplary and explanatory and do not limit the invention claimed.

Brief Description of the Drawings

[0018] The accompanying drawings illustrate several embodiments of the disclosed methods and compositions and, together with the specification, are used to interpret the principles of the disclosed methods and compositions. [Figure 1] This is a diagram of Scheme 2 (late peptide cyclization using cyclic ketones). [Figure 2] This is a diagram illustrating an example of late peptide cyclization using different cyclic ketones. [Figure 3] This is a diagram illustrating an example of late-stage peptide cyclization of a natural peptide using a cyclic ketone. [Figure 4] This is a diagram of Scheme 3 (late peptide cyclization using acetone). [Figure 5] This is a diagram illustrating an example of late-stage peptide cyclization of a natural peptide using acetone. [Figure 6] The UV (190-400 nm) and MS (250-3000 m / z) traces of 2a were obtained by UPLC-MS analysis (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C60H79N13O14S2 were [M+H]+m / z=1271.4, measured value 1270.5; calculated value: [M+2H]2+m / z=636.2, measured value 636.1. [Figure 7] The UV (190-400 nm) and MS (250-3000 m / z) traces of 2b were obtained by UPLC-MS analysis (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated value for C30H36N6O6S2 was [M+H]+m / z=641.2, and the measured value was 641.3. [Figure 8] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from UPLC-MS analysis of 2c (10-50% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C59H73N15O12S2 were [M+H]+m / z=1249.4, measured value 1248.9; calculated value: [M+2H]2+m / z=625.2, measured value 625.0. [Figure 9] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from 2d UPLC-MS analysis (10-50% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C66H83N15O15S2 were [M+H]+m / z=1391.5, measured value 1391.9; calculated value: [M+2H]2+m / z=696.3, measured value 696.7. [Figure 10] The following are UV (190-400 nm) and MS (250-3000 m / z) traces from UPLC-MS analysis of 2e (10-50% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C62H76N18O9S2 are [M+2H]2+m / z=641.7, measured value 641.9; calculated value: [M+3H]3+m / z=428.1, measured value 428.2. [Figure 11] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from UPLC-MS analysis of 2f (15-60% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C67H77N11O10S2 were [M+H]+m / z=1172.3, measured value 1171.7; calculated value: [M+2H]2+m / z=586.7, measured value 586.6. [Figure 12] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from 2 g of UPLC-MS analysis (20-70% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C67H77N11O10S2 were [M+H]+m / z=1261.5, measured value 1260.8; calculated value: [M+2H]2+m / z=631.2, measured value 631.3. [Figure 13]UV (190-400 nm) and MS (300-2000 m / z) traces were obtained by UPLC-MS analysis over 2 hours (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C51H77N13O14S2 were [M+H]+m / z=1161.3, measured value 1160.6; the calculated value was [M+2H]2+m / z=581.1, measured value 581.1. [Figure 14] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from UPLC-MS analysis of 2i (10-50% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C52H80N14O14S2 were [M+H]+m / z=1190.4, measured value 1189.8; calculated value: [M+2H]2+m / z=595.7, measured value 595.6. [Figure 15] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from 2j via UPLC-MS analysis (10-50% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). ESI-MS calculated values ​​for C55H82N14O14S2: [M+Na]+m / z=1249.9, measured value 1249.9; calculated value: [M+H]+m / z=1228.4, measured value 1227.8; calculated value: [M+2H]2+m / z=614.7, measured value 614.6. [Figure 16] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained by 2k UPLC-MS analysis (10-60% CH3CN / H2O gradient containing 0.1% TFA, 8 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C57H89N17O14S2 were [M+H]+m / z=1301.5, measured value 1300.8; calculated value: [M+2H]2+m / z=651.2, measured value 651.1. [Figure 17]UPLC-MS analysis of cyclo-(AcHN-GCYIQNCPLG-CONH2) yielded UV (190-400 nm) and MS (250-3000 m / z) traces (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). ESI-MS calculations for C50H77N13O14S2: [M+H]+m / z=1149.3, measured value 1148.6; calculation: [M+2H]2+m / z=575.1, measured value 575.1. [Figure 18] The UV (190-400 nm) and MS (250-3000 m / z) traces of 3b were obtained by UPLC-MS analysis (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C56H81N15O15S2 were [M+H]+m / z=1269.4, measured value 1270.2; calculated value: [M+2H]2+m / z=635.2, measured value 635.6. [Figure 19] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from UPLC-MS analysis of 3c (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C52H74N18O9S2 are: [M+H+TFA]+m / z=1274.4, measured value 1273.7; calculated value: [M+H]+m / z=1160.3, measured value 1159.5; calculated value: [M+2H]2+m / z=580.7, measured value 580.6; calculated value: [M+3H]3+m / z=387.4, measured value 387.6. [Figure 20] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained by 3D ULC-MS analysis (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C57H75N11O10S2 were [M+H]+m / z=1139.4, measured value 1138.6; calculated value: [M+2H]2+m / z=570.2, measured value 570.0. [Figure 21]The 3e sample was analyzed by UPLC-MS using UV (190-400 nm) and MS (300-2000 m / z) traces (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C52H72N10O10S2 were [M+H]+m / z=1062.32, measured value 1061.7; the calculated value was [M+2H]2+m / z=531.6, measured value 531.7. [Figure 22] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from UPLC-MS analysis of 3f (5-95% CH3CN / H2O gradient containing 0.1% TFA, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C66H124N24O13S2 were: [M+2H]2+m / z=763.9, measured value 764.0; calculated value: [M+3H]3+m / z=509.6, measured value 509.8; calculated value: [M+3H]3+m / z=382.5, measured value 382.8. [Figure 23] UV (190-400 nm) and MS (250-3000 m / z) traces were obtained from 3 g of UPLC-MS analysis (containing 0.1% TFA, 5-95% CH3CN / H2O gradient, 5 minutes, flow rate 0.4 mL / min). The ESI-MS calculated values ​​for C79H110N18O19S2 were: [M+H]+m / z=1680.9, measured value 1680.3; calculated value: [M+2H]2+m / z=840.9, measured value 840.9; calculated value: [M+3H]3+m / z=560.9, measured value 561.0. [Modes for carrying out the invention]

[0019] The disclosed methods and compositions can be more easily understood by referring to the detailed description of the following specific embodiments and the examples contained herein, as well as the drawings and the descriptions before and after them.

[0020] This application discloses a method for constructing disulfide mimetic compounds for late-stage cyclization of peptides. Natural disulfide bonds are unstable under reducing conditions, thus limiting their broad applications. The disclosed late-stage cyclization reaction involves thioacetalization of a peptide containing two cysteine ​​residues, forming a thioacetal group that is stable under acidic, basic, and reducing conditions. Compared to existing methods, the disclosed peptide thioacetalization can generate novel disulfide mimetic compounds. A wide range of cyclic ketones, and even acetone, can be used as crosslinking agents in this reaction. Since trifluoroacetic acid (TFA) can function as both a catalyst and the sole solvent, virtually all peptide sequences can be used in the disclosed method.

[0021] The disclosed method offers numerous advantages and merits, including the use of TFA as a robust catalyst and the only solvent required for the reaction, the readily available availability of acetone and a variety of cyclic ketones as reactants, good chemoselectivity and tolerance of natural peptides to this reaction, and the fact that the resulting crosslinks are structural mimics of disulfides.

[0022] To overcome the drawbacks of natural disulfide bonds, various stapling methods based on highly reactive and selective cysteine ​​peptides for forming disulfide bond mimics already exist. Stapling is achieved using symmetric linkers such as dichloroacetone (DCA), dichloroacetophenone, dibromobenzyl linkers, or two-lead-substituting aryl linkers that act via S-alkylation or S-arylation. Recently, Cramer et al. reacted a CH2I2 reagent with a cysteine ​​peptide to form a thiocarbenium ion intermediate, which was then converted to a methylenethioacetal, thereby eliminating the reductive instability of the disulfide group and improving structural stability. In contrast, the disclosed method allows cyclic ketones and acetone to be used as crosslinking agents for late peptide cyclization based on cysteine ​​residues under trifluoroacetic acid (TFA) conditions, generating a series of thioacetal disulfide mimics with good chemoselectivity and resistance to natural peptides.

[0023] A typical general procedure for forming a disulfide mimetic by performing late cyclization using a cyclic ketone or acetone is shown in Scheme 1, where R 1 ~R 3 We define it as follows: R1 and R3 are independently H, Bn, an allyl group, or a C1-C6 alkyl group. R2 is a monosubstituted or polysubstituted fluorenone, such as H, a halogen, CN, CF3, NO2, an alkyl group, an alkoxy group, or an aryl group.

[0024] Scheme 1. Late cyclization for forming disulfide mimetic compounds using cyclic ketones and acetone. JPEG2026521678000022.jpg29147 JPEG2026521678000023.jpg29147

[0025] Scheme 2 (Figure 1) shows an exemplary general method for late peptide cyclization. A purified native peptide sequence after SPPS, such as linear vasopressin, terlipressin, cetomelanotide, oxytocin, or lanreotide, is dissolved in trifluoroacetic acid (TFA) to a final concentration of 10 mM. A 200 mM cyclic ketone (20.0 equivalents) is added to the solution, and the reaction mixture is stirred at room temperature for approximately 8 hours. After the reaction is complete, the solvent is blown off under a stream of N2, followed by decantation of diethyl ether. The residue is dissolved in a mixed solvent of acetonitrile (ACN) and water (H2O), and then purified by preparative HPLC. After lyophilization, the corresponding product is obtained. Here, a detailed synthetic procedure for late peptide modification using a cyclic ketone is described below as an example.

[0026] Scheme 3 (Figure 4) shows another exemplary general method for late peptide cyclization using acetone. Purified native peptide sequences after SPPS, such as terlipressin, cetomelanotide, lanreotide, octreotide, bactenesin, and somatostatin, are dissolved in a mixed solvent of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture is stirred at room temperature for approximately 8 hours. After the reaction is complete, the solvent is blown off under a stream of N2, and the crude peptide product is precipitated with diethyl ether. The residue is dissolved in a mixed solvent of acetonitrile (ACN) and water (H2O), and then purified by preparative HPLC. After lyophilization, the target product is obtained. Here, a detailed synthetic procedure for late peptide modification using cyclic ketones is described below as an example.

[0027] This application discloses a method for the late cyclization of a peptide and reagents used in the method. Generally, the method comprises maintaining a reaction mixture at a temperature sufficient to form a product for a sufficient amount of time. Generally, the reaction mixture comprises a peptide containing two or more cysteine ​​residues, a cyclic ketone reagent or acetone, and a solvent. Generally, the product comprises a thioacetalized peptide, in which two cysteine ​​residues of the peptide are linked by thioacetalization.

[0028] In some forms, peptides are linear or cyclic (including monocyclic and bicyclic structures). In some forms, the peptide is a random peptide or a peptide drug. In some forms, the peptide is formed from natural amino acids.

[0029] In some forms, cyclic ketone reagents or acetone are cyclic ketone reagents having the following structure: JPEG2026521678000024.jpg3024 Formula I In the formula, R and R' are independently alkyl groups (e.g., C1-C6 alkyl groups), and TIFF2026521678000025.tif212 is absent, or R and R' together form a cyclic moiety A, where A is a cycloalkyl group, cycloalkenyl group, cycloalkynyl group, aryl group, polyaryl group, heteroaryl group, heteropolyaryl group, or heterocyclic group, where R'' represents hydrogen or a substituent on the cyclic moiety A, and R'' is, whenever it appears, independently a benzyl group, an allyl group, an alkyl group (e.g., a C1-C6 alkyl group), a halogen, -CN, -CF3, -NO2, an alkoxy group, or an aryl group (e.g., a phenyl group), and n is an integer between 0 and 10, 0 and 8, 0 and 6, 0 and 4, or 0 and 2. If R'' is an alkyl group, the alkyl group may be substituted or unsubstituted, and the substituent, if present, may be any substituent disclosed herein, e.g., a halogen, an azide group, or an alkynyl group (e.g., -CCH or -CH2CCH).

[0030] In some forms, R and R' are independently alkyl groups such as methyl groups. In some forms, R and R' are independently alkyl groups such as methyl groups, and TIFF2026521678000026.tif212 does not exist. In some forms, A is cyclobutane, azetidine, cyclopentane, fluorenone, cyclohexane, or piperidine.

[0031] In some forms, the solvent is trifluoroacetic acid. In some forms, trifluoroacetic acid acts as a catalyst and is the only solvent in the reaction mixture.

[0032] In some forms, the reaction mixture is maintained at a temperature of 20°C to 35°C, such as approximately 30°C, for a period of up to 1 hour, up to 2 hours, up to 3 hours, 10 minutes to 1 hour, 20 minutes to 2 hours, or 30 minutes to 3 hours.

[0033] In some forms, the molar ratio of peptide to cyclic ketone reagent (peptide:cyclic ketone reagent) is between 0.1 and 1, for example, about 0.2.

[0034] In some forms, cyclic ketone reagents have one of the following structures: JPEG2026521678000027.jpg27118 Here, R 1 and R 3 The elements are H, benzyl group, allyl group, alkyl group (e.g., C1-C6 alkyl group), and each R 2 R1, R2, R3, R3 are independently H, a benzyl group, an allyl group, an alkyl group (e.g., a C1-C6 alkyl group), a halogen, -CN, -CF3, -NO2, an alkoxy group, or an aryl group. If any of R1-R3 is an alkyl group, the alkyl group may be substituted or unsubstituted, and the substituent, if present, may be any substituent disclosed herein, such as a halogen, an azide group, or an alkynyl group (e.g., -CCH or -CH2CCH).

[0035] In some forms, thioacetalized peptides have one of the following structures: JPEG2026521678000028.jpg5891, JPEG2026521678000029.jpg6980, JPEG2026521678000030.jpg6993, JPEG2026521678000031.jpg6999, JPEG2026521678000032.jpg5495, JPEG2026521678000033.jpg6368, JPEG2026521678000034.jpg6194, JPEG2026521678000035.jpg5889, JPEG2026521678000036.jpg5889, JPEG2026521678000037.jpg5990, JPEG2026521678000038.jpg5986, JPEG2026521678000039.jpg6299, JPEG2026521678000040.jpg6185, JPEG2026521678000041.jpg6873, JPEG2026521678000042.jpg6268, JPEG2026521678000043.jpg69126, or JPEG2026521678000044.jpg8884.

[0036] The disclosed methods and compositions are not limited to specific synthesis methods, analytical techniques, or reagents unless otherwise specified, and should be understood to be modifiable. The terms used herein are for the purpose of describing specific embodiments only and are not intended to be limiting.

[0037] As used herein, the term “substitution” refers to all permissible substituents of the compounds or functional groups described herein. In its broadest sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, but are not limited to, halogens, hydroxyl groups, or any other organic group in a linear, branched, or cyclic structural form containing any number of carbon atoms (preferably 1 to 14 carbon atoms) and optionally containing one or more heteroatoms such as oxygen, sulfur, or nitrogen. Typical substituents include substituted or unsubstituted alkyl groups, substituted or unsubstituted alkenyl groups, substituted or unsubstituted alkynyl groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted phenyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, substituted or unsubstituted polyaryl groups, substituted or unsubstituted polyheteroaryl groups, substituted or unsubstituted aralkyl groups, halogens, hydroxyl groups, alkoxy groups, phenoxy groups, alloxy groups, silyl groups, thiol groups, alkylthio groups, substituted alkylthio groups, phenylthio groups, arylthio groups, cyano groups, isocyano groups, nitro groups, substituted or unsubstituted carbonyl groups, carboxyl groups, amino groups, amide groups, oxo, sulfinyl groups, sulfonyl groups, sulfonic acids, phosphonium groups, phosphanyl groups, phosphoryl groups, phosphonyl groups, and amino acids. Such substituted or unsubstituted alkyl groups, substituted or unsubstituted alkenyl groups, substituted or unsubstituted alkynyl groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted phenyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, substituted or unsubstituted polyaryl groups, substituted or unsubstituted polyheteroaryl groups, substituted or unsubstituted aralkyl groups, halogens, hydroxyl groups, alkoxy groups, phenoxy groups, alloxy groups, silyl groups, thiol groups, alkylthio groups, substituted alkylthio groups, phenylthio groups, arylthio groups, cyano groups, isocyano groups, nitro groups, substituted or unsubstituted carbonyl groups, carboxyl groups, amino groups, amide groups, oxo, sulfinyl groups, sulfonyl groups, sulfonic acids, phosphonium groups, phosphanyl groups, phosphoryl groups, phosphonyl groups, and amino acids may be further substituted.

[0038] Heteroatoms such as nitrogen may have any acceptable substituents on the organic compounds described herein that satisfy the hydrogen substituent and / or the valence of the heteroatom. It should be understood that the terms “substituted” or “substituted” implicitly include the condition that such substitutions result in a stable compound that does not undergo spontaneous changes such as rearrangement, cyclization, or elimination, and that such substitutions are subject to the acceptable valences of the substituted atom and substituent.

[0039] As used herein, "alkyl group" refers to a saturated aliphatic radical, including linear alkyl groups, branched alkyl groups, and cycloalkyl groups (alicyclic groups). In some forms, linear or branched alkyl groups have 30 or fewer (for example, a linear group with C1-C12 30 , branch chain C3~C 30 ), have 20 or fewer carbon atoms, 15 or fewer carbon atoms, or 10 or fewer carbon atoms. Alkyl groups include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, t-butyl group, pentyl group, hexyl group, heptyl group, octyl group, decyl group, tetradecyl group, hexadecyl group, eicosyl group, tetracosyl group, etc. Similarly, cycloalkyl groups are non-aromatic carbon-based rings consisting of at least three carbon atoms, and examples include non-aromatic monocyclic or non-aromatic polycyclic rings containing 3 to 30 carbon atoms, 3 to 20 carbon atoms, or 3 to 10 carbon atoms in their ring structure, and having 5, 6, or 7 carbon atoms in their ring structure. Cycloalkyl groups containing polycyclic ring systems may have two or more non-aromatic rings (i.e., "condensed cycloalkyl rings") in which two adjacent rings share two or more carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctanyl groups.

[0040] A "substituted alkyl group" refers to an alkyl group having one or more substituents that substitute for hydrogen atoms on one or more carbon atoms of a hydrocarbon skeleton. Such substituents may be any of the above substituents, and include, for example, halogens (e.g., fluorine, chlorine, bromine, or iodine), hydroxyl groups, carbonyl groups (e.g., carboxyl groups, alkoxycarbonyl groups, formyl groups, or acyl groups), thiocarbonyl groups (e.g., thioesters, thioacetates, or thioformates), aryl groups, alkoxy groups, aralkyl groups, phosphonium groups, phosphanyl groups, phosphonyl groups, phosphoryl groups, phosphates, phosphonates, phosphinates, amino groups, amide groups, amidines, imines, cyano groups, nitro groups, azide groups, oxo groups, sulfhydryl groups, thiol groups, alkylthio groups, silyl groups, sulfinyl groups, sulfates, sulfonates, sulfamoyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, aromatic or heteroaromatic moieties. -NRR' (where R and R' are independently hydrogen, an alkyl group, or an aryl group, and the nitrogen atom is optionally quaternized); -SR (where R is a phosphonyl group, a sulfinyl group, a silyl group, hydrogen, an alkyl group, or an aryl group); -CN; -NO2; -COOH; carboxylate salts; -COR, -COOR, or -CON(R)2 (where R is hydrogen, an alkyl group, or an aryl group); imino groups, silyl groups, ethers, haloalkyl groups (e.g., -CF3, -CH2-CF3, -CCl3); -CN, -NCOCOCH2CH2; -NCOCOCHCH; and -NCS; and combinations thereof.

[0041] Those skilled in the art should understand that a substituted portion on a hydrocarbon chain can, if appropriate, be substituted itself. For example, substituents on a substituted alkyl group may include halogens, hydroxyl groups, nitro groups, thiol groups, amino groups, aralkyl groups, azide groups, imino groups, amide groups, phosphonium groups, phosphanyl groups, phosphoryl groups (including phosphonates and phosphinates), oxo, sulfonyl groups (including sulfates, sulfonamides, sulfamoyl groups and sulfonates), silyl groups, and ethers, alkylthio groups, carbonyl groups (including ketones, aldehydes, carboxylates and esters), haloalkyl groups, -CN, etc. Cycloalkyl groups can be substituted in a similar manner.

[0042] Unless otherwise specified, the term "lower alkyl group" as used herein refers to the alkyl group defined above, having 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, in its skeletal structure. Similarly, "lower alkenyl group" and "lower alkynyl group" have similar chain lengths.

[0043] As used herein, “heteroalkyl group” refers to a linear, branched, or cyclic carbon-containing alkyl radical or a combination thereof, which contains at least one heteroatom on its carbon skeleton. Suitable heteroatoms include, but are not limited to, O, N, Si, P, and S, of which nitrogen, phosphorus, and sulfur atoms may be optionally oxidized, and nitrogen heteroatoms may be optionally quaternized. For example, the term “heteroalkyl group” is a cycloalkyl group as defined above, in which at least one carbon atom of the ring is substituted with a heteroatom, and the heteroatom may be, for example, nitrogen, oxygen, sulfur, or phosphorus.

[0044] As used herein, the term "alkenyl group" refers to a hydrocarbon group having a structural formula containing 2 to 24 carbon atoms and at least one carbon-carbon double bond. Alkenyl groups include linear alkenyl groups, branched alkenyl groups, and cycloalkenyl groups. A cycloalkenyl group is a non-aromatic carbon ring consisting of at least 3 carbon atoms and at least 1 carbon-carbon double bond, for example, a non-aromatic monocyclic or non-aromatic polycyclic ring whose ring structure contains 3 to 30 carbon atoms and at least 1 carbon-carbon double bond, 3 to 20 carbon atoms and at least 1 carbon-carbon double bond, or 3 to 10 carbon atoms and at least 1 carbon-carbon double bond, and whose ring structure contains 5, 6, or 7 carbon atoms and at least 1 carbon-carbon double bond. Cycloalkenyl groups containing polycyclic ring systems may have two or more non-aromatic rings, where two or more carbons are shared by two adjacent rings (i.e., “condensed cycloalkenyl rings”) and contain at least one carbon-carbon double bond. Asymmetric structures such as (AB)C=C(C'D) are intended to include both E and Z isomers. This can be inferred in the structural formulas in which the asymmetric alkenes of this specification exist, or it may be explicitly indicated by the bond symbol C. As used throughout the specification, examples, and claims, the term “alkenyl group” is intended to include both “unsubstituted alkenyl groups” and “substituted alkenyl groups,” the latter referring to alkenyl group moieties having one or more substituents substituting hydrogens on one or more carbons of a hydrocarbon skeleton. The term “alkenyl group” also includes “heteroalkenyl groups.”

[0045] The term "substituted alkenyl group" refers to an alkenyl group portion having one or more substituents that substitute one or more hydrogen atoms on one or more carbon atoms of a hydrocarbon skeleton. Such substituents may be any of the above substituents, for example, halogens, azides, alkyl groups, aralkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, hydroxyl groups, carbonyl groups (e.g., carboxyl groups, alkoxycarbonyl groups, formyl groups, or acyl groups), silyl groups, ethers, esters, thiocarbonyl groups (e.g., thioesters, thioacetates, or thioformates), alkoxy groups, phosphonium groups, phosphanyl groups, phosphoryl groups, phosphates, phosphonates, phosphinates, amino groups (e.g., quaternized amino groups), amide groups, amidines, imines, cyano groups, nitro groups, azide groups, oxo groups, sulfhydryl groups, alkylthio groups, sulfates, sulfonates, sulfamoyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, alkylaryl groups, haloalkyl groups, -CN groups, aryl groups, heteroaryl groups, polyaryl groups, polyheteroaryl groups, and combinations thereof.

[0046] As used herein, “heteroalkenyl group” refers to a linear, branched, or cyclic carbon-containing alkenyl radical or combination thereof containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, and S, of which nitrogen, phosphorus, and sulfur atoms may be optionally oxidized, and nitrogen heteroatoms may be optionally quaternized. For example, the term “heterocycloalkenyl group” refers to a cycloalkenyl group in which at least one carbon atom of the ring is substituted with a heteroatom, such as, but is not limited to, nitrogen, oxygen, sulfur, or phosphorus.

[0047] As used herein, the term "alkynyl group" refers to a hydrocarbon group having a structural formula containing 2 to 24 carbon atoms and at least one carbon-carbon triple bond. Alkynyl groups include linear alkynyl groups, branched alkynyl groups, and cycloalkynyl groups. A cycloalkynyl group is a non-aromatic carbon ring consisting of at least 3 carbon atoms and at least one carbon-carbon triple bond, for example, a non-aromatic monocyclic or non-aromatic polycyclic ring whose ring structure contains 3 to 30 carbon atoms and at least one carbon-carbon triple bond, 3 to 20 carbon atoms and at least one carbon-carbon triple bond, or 3 to 10 carbon atoms and at least one carbon-carbon triple bond, and whose ring structure contains 5, 6, or 7 carbon atoms and at least one carbon-carbon triple bond. A cycloalkynyl group containing a polycyclic ring system may have two or more non-aromatic rings, where two or more carbons are shared by two adjacent rings (i.e., a “condensed cycloalkynyl ring”) and contains at least one carbon-carbon triple bond. Asymmetric structures such as (AB)C≡C(C'D) are intended to encompass both E and Z isomers. This can be inferred in the structural formulas in which the asymmetric alkynes exist herein, or it may be explicitly indicated by the bond symbol C. As used throughout the specification, examples, and claims, the term “alkynyl group” is intended to include both “unsubstituted alkynyl groups” and “substituted alkynyl groups,” the latter referring to an alkynyl group moiety having one or more substituents substituting hydrogens on one or more carbons of a hydrocarbon skeleton. The term “alkynyl group” also includes “heteroalkynyl groups.”

[0048] A "substituted alkynyl group" refers to an alkynyl group portion that has one or more substituents that substitute for one or more hydrogen atoms on one or more carbon atoms of a hydrocarbon skeleton. Such substituents may be any of the above substituents, for example, halogens, azides, alkyl groups, aralkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, hydroxyl groups, carbonyl groups (e.g., carboxyl groups, alkoxycarbonyl groups, formyl groups, or acyl groups), silyl groups, ethers, esters, thiocarbonyl groups (e.g., thioesters, thioacetates, or thioformates), alkoxy groups, phosphonium groups, phosphanyl groups, phosphoryl groups, phosphates, phosphonates, phosphinates, amino groups (e.g., quaternized amino groups), amide groups, amidines, imines, cyano groups, nitro groups, azide groups, sulfhydryl groups, alkylthio groups, sulfates, sulfonates, sulfamoyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, alkylaryl groups, haloalkyl groups, -CN groups, aryl groups, heteroaryl groups, polyaryl groups, polyheteroaryl groups, and combinations thereof.

[0049] As used herein, “heteroalkynyl group” refers to a linear, branched, or cyclic carbon-containing alkynyl radical or combination thereof containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, and S, of which nitrogen, phosphorus, and sulfur atoms may be optionally oxidized, and nitrogen heteroatoms may be optionally quaternized. For example, the term “heterocycloalkynyl group” refers to a cycloalkynyl group in which at least one carbon atom of the ring is substituted with a heteroatom, such as, but is not limited to, nitrogen, oxygen, sulfur, or phosphorus.

[0050] As used herein, the term "aryl group" refers to a C5-C5 group. 26 This refers to a system of aromatic or condensed aromatic rings. Examples of aromatic groups include benzene, naphthalene, anthracene, phenanthrene, chrysene, pyrene, corannulene, and coronene.

[0051] The "substituted aryl group" refers to an aryl group in which one or more hydrogen atoms on one or more aromatic rings are substituted by one or more substituents. The substituents include, but are not limited to, halogen, azide, alkyl group, aralkyl group, alkenyl group, alkynyl group, cycloalkyl group, hydroxyl group, alkoxy group, carbonyl group (such as ketone, aldehyde, carboxyl group, alkoxycarbonyl group, formyl group or acyl group), silyl group, ether, ester, thiocarbonyl group (such as thioester, thioacetate or thioformate), alkoxy group, phosphoryl group, phosphate, phosphonate, phosphinate, amino group (or quaternized amino group), amide group, amidine, imine, cyano group, nitro group, azide group, sulfhydryl group, imino group, alkylthio group, sulfate, sulfonate, sulfamoyl group, sulfoxide, sulfonamide group, sulfonyl group, heterocyclic group, alkylaryl group, haloalkyl group (such as CF3, -CH2-CF3, -CCl3), -CN, aryl group, heteroaryl group, and combinations thereof.

[0052] The "heterocyclic ring" and the "heterocyclic group" are used interchangeably and refer to a cyclic radical linked via a carbon or nitrogen atom on a non-aromatic monocyclic or polycyclic ring containing 3 to 30 ring atoms, 3 to 20 ring atoms, 3 to 10 ring atoms or 5 to 6 ring atoms. Each ring contains carbon and 1 to 4 heteroatoms, and each heteroatom is independently selected from non-peroxide oxygen, sulfur and N(Y), where Y is absent or H, O, C1~C 10The group is an alkyl group, a phenyl group, or a benzyl group, and optionally contains 1 to 3 double bonds, and optionally is substituted with one or more substituents. By definition, heterocyclic groups are different from heteroaryl groups. Heterocyclic groups can be heterocycloalkyl groups, heterocycloalkenyl groups, heterocycloalkynyl groups, etc., for example, piperazinyl group, piperidinyl group, piperidonyl group, 4-piperidonyl group, dihydrofluoro[2,3-b]tetrahydrofuran, morpholinyl group, piperazinyl group, piperidinyl group, piperidonyl group, 4-piperidonyl group, piperonyl group, pyranyl group, 2H-pyrrolyl group, 4H-quinolidinyl group, quinuclidinyl group, tetrahydrofuranyl group, or 6H-1,2,5-thiadiadinyl. Heterocyclic groups can optionally be substituted with one or more substituents as defined above for alkyl and aryl groups.

[0053] The term "heteroaryl group" refers to a C5-C ring structure in which one or more carbon atoms on one or more aromatic ring structures are already substituted by heteroatoms. 26This refers to a heteroaryl group consisting of aromatic or fused aromatic rings. Suitable heteroatoms include, but are not limited to, oxygen, sulfur, and nitrogen. Examples of heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, tetrazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.Examples of heteroaryl rings include benzimidazolyl group, benzofuranyl group, benzothiofuranyl group, benzothiophenyl group, benzoxazolyl group, benzoxazolinyl group, benzthiazolyl group, benztriazolyl group, benztetrazolyl group, benzisoxazolyl group, benzisothiazolyl group, benzimidazolinyl group, carbazolyl group, 4aH-carbazolyl group, carbolinyl group, chromanyl group, chromenyl group, synnolinyl group, decahydroquinolinyl group, 2H,6H-1,5,2-dithiadinyl group, furanyl group, and f Lazanyl group, imidazolidinyl group, imidazolinyl group, imidazolyl group, 1H-indazolyl group, indolenyl group, indolinyl group, indolidinyl group, indolyl group, 3H-indolyl group, isatinoyl group, isobenzofuranyl group, isochromanyl group, isoindazolyl group, isoindolinyl, isoindolyl group, isoquinolinyl group, isothiazolyl group, isoxazolyl group, methylenedioxyphenyl group, naphthilidinyl group, octahydroisoquinolinyl group, 1,2,3-oxadiazolyl group, 1,2,4-oxadiazo Lyl group, 1,2,5-oxadiazolyl group, 1,3,4-oxadiazolyl group, oxazolidinyl group, oxazolyl group, oxyindolyl group, pyrimidinyl group, phenanthrolinyl, phenanthrolinyl, phenazinyl group, phenothiazinyl group, phenoxathinyl group, phenoxazinyl group, phthalazinyl group, pteridinyl group, purinyl group, pyrazinyl group, pyrazolidinyl group, pyrazolinyl group, pyrazolyl group, pyridadinyl group, pyridoxazole group, pyridoimidazole group, pyridothiazole group, pyridinyl group, pyridyl group This includes, but is not limited to, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl groups. One or more rings can be substituted as defined below for "substituted heteroaryl groups".

[0054] The term "substituted heteroaryl group" refers to a heteroaryl group in which one or more hydrogen atoms on one or more heteroaromatic rings are substituted by one or more substituents, and the substituents include halogens, azides, alkyl groups, aralkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, hydroxyl groups, alkoxy groups, carbonyl groups (e.g., ketones, aldehydes, carboxyl groups, alkoxycarbonyl groups, formyl groups, or acyl groups), silyl groups, ethers, esters, and thiocarbonyl groups (e.g., thioesters, thioacetates, or thio This includes, but is not limited to, formates, alkoxy groups, phosphoryl groups, phosphates, phosphonates, phosphinates, amino groups (or quaternized amino groups), amide groups, amidines, imines, cyano groups, nitro groups, azide groups, sulfhydryl groups, imino groups, alkylthio groups, sulfates, sulfonates, sulfamoyl groups, sulfoxides, sulfonamide groups, sulfonyl groups, heterocyclic groups, alkylaryl groups, haloalkyl groups (e.g., CF3, -CH2-CF3, -CCl3), -CN, aryl groups, heteroaryl groups, and combinations thereof.

[0055] The term "polyaryl group" refers to a chemical moiety containing two or more condensed aryl groups. When involving two or more condensed heteroaryl groups, the chemical moiety may be called a "polyheteroaryl group."

[0056] A "substituted polyaryl group" refers to a polyaryl group in which one or more aryl groups are substituted by one or more substituents, and such substituents include, but are not limited to, halogens, azides, alkyl groups, aralkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, hydroxyl groups, carbonyl groups (e.g., carboxyl groups, alkoxycarbonyl groups, formyl groups, or acyl groups), silyl groups, ethers, esters, thiocarbonyl groups (e.g., thioesters, thioacetates, or thioformates), alkoxy groups, phosphoryl groups, phosphates, phosphonates, phosphinates, amino groups (or quaternized amino groups), amide groups, amidines, imines, cyano groups, nitro groups, azide groups, sulfhydryl groups, alkylthio groups, sulfates, sulfonates, sulfamoyl groups, sulfoxides, sulfonamide groups, sulfonyl groups, heterocyclic groups, alkylaryl groups, haloalkyl groups, -CN groups, aryl groups, heteroaryl groups, and combinations thereof. When involving a polyheteroaryl group, the chemical moiety in question may be called a "substituted polyheteroaryl group."

[0057] The term "cyclic ring" or "cyclic group" refers to a substituted or unsubstituted monocyclic ring or a substituted or unsubstituted polycyclic ring (e.g., one formed from a monocyclic or fused ring system), such as a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted cycloalkenyl group, a substituted or unsubstituted cycloalkynyl group, or a substituted or unsubstituted heterocyclic group, which has 3 to 30 carbon atoms, as geometric constraints allow. The substituted cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, and heterocyclic groups are substituted as defined above for alkyl groups, alkenyl groups, alkynyl groups, and heterocyclic groups, respectively.

[0058] As used herein, the term "aralkyl group" refers to an aryl or heteroaryl group having an alkyl, alkynyl, or alkenyl group as defined above, which is linked to an aromatic group, and the aromatic group is, for example, an aryl group, a heteroaryl group, a polyaryl group, or a polyheteroaryl group. An example of an aralkyl group is the benzyl group.

[0059] The term "thiol group" is used interchangeably and is denoted as -SR, where R may be hydrogen, a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group, a substituted or unsubstituted alkynyl group, a substituted or unsubstituted heterocyclic group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted aralkyl group (e.g., a substituted or unsubstituted alkylaryl group, a substituted or unsubstituted aralkyl group, etc.), a substituted or unsubstituted polyaryl group, a substituted or unsubstituted polyheteroaryl group, a substituted or unsubstituted carbonyl group, a phosphonium group, a phosphanyl group, an amide group, an amino group, an alkoxy group, an oxo, a phosphonyl group, a sulfinyl group, or a silyl group, as described above. Such substituents may be any of the above substituents, such as halogens, azides, alkyl groups, aralkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, hydroxyl groups, carbonyl groups (e.g., carboxyl groups, alkoxycarbonyl groups, formyl groups, or acyl groups), silyl groups, ethers, esters, thiocarbonyl groups (e.g., thioesters, thioacetates, or thioformates), alkoxy groups, phosphoryl groups, phosphates, phosphonates, phosphinates, amino groups (e.g., quaternized amino groups), amide groups, amidines, imines, cyano groups, nitro groups, azide groups, sulfhydryl groups, alkylthio groups, sulfates, sulfonates, sulfamoyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, alkylaryl groups, haloalkyl groups, -CN groups, aryl groups, heteroaryl groups, polyaryl groups, polyheteroaryl groups, and combinations thereof.

[0060] The disclosed compounds and substituents may independently have two or more of the groups listed above. For example, if the compound or substituent is a linear alkyl group, one of the hydrogen atoms of the alkyl group may be substituted with a hydroxyl group, an alkoxy group, etc. Depending on the selected group, the first group may be incorporated into the second group or selectively pendanted (i.e., linked) to the second group. For example, in the expression "alkyl group containing an ester group," the ester group may be incorporated into the alkyl group skeleton, or the ester may be linked to the alkyl group skeleton. The properties of the selected group determine whether the first group is incorporated into or linked to the second group.

[0061] Compounds and substituents can be independently substituted by substituents as defined above.

[0062] A numerical range individually discloses all numerical values ​​that can reasonably be included within that range, as well as any partial ranges and combinations of partial ranges that are included therein. For example, within a given carbon range of C3 to C9, the range further discloses C3, C4, C5, C6, C7, C8, and C9, any partial range between these numerical values ​​(e.g., C4 to C6), and all possible combinations of possible ranges between these values. In yet another example, a given temperature range may be approximately 25°C to 30°C, where the range independently discloses temperatures selected from approximately 25°C, 26°C, 27°C, 28°C, 29°C, and 30°C, any range between these numerical values ​​(e.g., 26°C to 28°C), and all possible combinations of ranges between these values.

[0063] The use of the term "approximately" is intended to indicate that the values ​​above and below the number being modified fall within a range of approximately ±10%. When the term "approximately" is used before a numerical range (i.e., approximately 1 to 5) or before a series of numbers (i.e., approximately 1, 2, 3, 4, etc.), it is intended to modify each number listed at both ends of the numerical range and / or the entire series of numbers, unless otherwise specified.

[0064] "Oxo" refers to =O.

[0065] Compounds and substituents can be independently substituted by substituents greater than or equal to those specified in the definition of "substitution".

[0066] The numerical range includes the range from 1 to 6. The range individually discloses all numerical values ​​that can reasonably be included within that range, as well as any partial ranges and combinations of partial ranges that are included therein. For example, within a given carbon range of C3 to C9, the range further discloses C3, C4, C5, C6, C7, C8, and C9, any partial range between these numerical values ​​(e.g., C4 to C6), and all possible combinations of possible ranges between these values. In yet another example, a given temperature range may be approximately 25°C to 30°C, where the range independently discloses temperatures selected from approximately 25°C, 26°C, 27°C, 28°C, 29°C, and 30°C, any range between these numerical values ​​(e.g., 26°C to 28°C), and all possible combinations of ranges between these values.

[0067] The use of the term "approximately" is intended to indicate that the values ​​above and below the number being modified fall within a range of approximately ±10%. When the term "approximately" is used before a numerical range (i.e., approximately 1 to 5) or before a series of numbers (i.e., approximately 1, 2, 3, 4, etc.), it is intended to modify each number listed at both ends of the numerical range and / or the entire series of numbers, unless otherwise specified.

[0068] A “metaphor” of a given compound refers to another compound that is structurally similar, functionally similar, or both to a particular compound. Structural similarity can be determined by any criterion known in the art, such as the Tanimoto coefficient, which provides a quantitative measure of similarity between two compounds based on their molecular descriptors. Preferably, the molecular descriptor is a two-dimensional property such as a fingerprint, topological index, or maximum common substructure, or a three-dimensional property such as overall shape or molecular field. The Tanimoto coefficient takes a value of 0 to 1 (including 0 and 1) for different molecular pairs and identical molecular pairs, respectively. If the Tanimoto coefficient between a compound and a designated compound is 0.5 to 1.0 (including 0.5 and 1.0), preferably 0.7 to 1.0 (including 0.7 and 1.0), and most preferably 0.85 to 1.0 (including 0.85 and 1.0), then the compound can be considered a metaphor of the designated compound. If a compound induces the same pharmacological, physiological, or both effects as a specified compound, the compound is functionally similar to the specified compound. “Similar” may also refer to modifications of the disclosed compound, including, but not limited to, products of hydrolysis, reduction, or oxidation. Hydrolysis, reduction, and oxidation reactions are well known in the art.

[0069] The disclosed compositions and methods can be further understood through the following numbered paragraphs.

[0070] 1. A method for late cyclization of peptides, The reaction mixture is maintained at a temperature sufficient to form the product for a sufficient amount of time. The reaction mixture comprises a peptide containing two or more cysteine ​​residues, a cyclic ketone reagent or acetone, and a solvent. The product comprises a thioacetalized peptide, and the thioacetalization process binds two cysteine ​​residues of the peptide.

[0071] 2. According to the method described in paragraph 1, the peptide is linear or cyclic (including monocyclic, bicyclic, etc.).

[0072] 3. According to the method described in paragraph 1 or 2, the peptide is a random peptide or a peptide drug.

[0073] 4. According to the method described in any of paragraphs 1 to 3, the peptide comprises natural amino acids.

[0074] 5. According to the method described in any of paragraphs 1 to 4, the cyclic ketone reagent is a cyclic ketone reagent having the following structure. JPEG2026521678000045.jpg3024 Formula I (In the formula, R and R' are independently alkyl groups, and TIFF2026521678000046.tif212 does not exist, or R and R' together form a ring portion A. Here, A is a cycloalkyl group, a cycloalkenyl group, a cycloalkynyl group, an aryl group, a polyaryl group, a heteroaryl group, a heteropolyaryl group, or a heterocyclic group. Here, R'' represents a hydrogen atom or a substituent on cyclic moiety A, and each time R'' appears, it is independently a benzyl group, an allyl group, an alkyl group (e.g., C1-C6 alkyl group), a halogen, -CN, or -CF. 3、 -NO2, an alkoxy group or an aryl group, and Here, n is an integer between 0 and 10, 0 and 8, 0 and 6, 0 and 4, or 0 and 2.

[0075] 6. According to the method described in paragraph 5, R and R' are independently alkyl groups such as methyl groups.

[0076] 7. According to the method described in paragraph 5 or 6, A is cyclobutane, azetidine, cyclopentane, fluorenone, cyclohexane, or piperidine.

[0077] 8. According to the method described in any of paragraphs 1 to 7, the solvent is trifluoroacetic acid.

[0078] 9. According to the method described in paragraph 8, trifluoroacetic acid acts as a catalyst and / or is the sole solvent in the reaction mixture.

[0079] 10. According to the method described in any paragraph of paragraphs 1 to 9, the reaction mixture is maintained at a temperature of 20°C to 35°C, such as about 30°C, for a period of up to 1 hour, up to 2 hours, up to 3 hours, 10 minutes to 1 hour, 20 minutes to 2 hours, or 30 minutes to 3 hours.

[0080] 11. According to the method described in any of paragraphs 1 to 10, the molar ratio of the peptide to the cyclic ketone reagent (peptide:cyclic ketone reagent) is 0.1 to 1, for example, about 0.2.

[0081] 12. According to the method described in any paragraph from paragraphs 1 to 11, the cyclic ketone reagent has one of the following structures: JPEG2026521678000047.jpg27118 (in the formula, R 1 and R 3 is H, benzyl group, allyl group, alkyl group (e.g., C1-C6 alkyl group), and Here, each R 2 These are independently H, a benzyl group, an allyl group, an alkyl group (e.g., a C1-C6 alkyl group), a halogen, -CN, -CF3, -NO2, an alkoxy group, or an aryl group.

[0082] 13. According to the method described in any paragraph from paragraphs 1 to 12, the thioacetalized peptide has one of the following structures: JPEG2026521678000048.jpg5891, JPEG2026521678000049.jpg6980, JPEG2026521678000050.jpg6993, JPEG2026521678000051.jpg6999, JPEG2026521678000052.jpg5495, JPEG2026521678000053.jpg6368, JPEG2026521678000054.jpg6194, JPEG2026521678000055.jpg5889, JPEG2026521678000056.jpg5889, JPEG2026521678000057.jpg5990, JPEG2026521678000058.jpg5986, JPEG2026521678000059.jpg6299, JPEG2026521678000060.jpg6185, JPEG2026521678000061.jpg6873, JPEG2026521678000062.jpg6268, JPEG2026521678000063.jpg69126, or JPEG2026521678000064.jpg8884. (Examples)

[0083] 1. AcHN-GCYIQNCPLG-CONH2+9-Fluorenone (2a) JPEG2026521678000065.jpg5891AcHN-GCYIQNCPLG-CONH2+9-Fluorenone(2a) The purified AcHN-GCYIQNCPLG-CONH2(1a) (11.1 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 200 mM 9-fluorenone (36.0 mg, 20.0 equivalents) was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (15-60%, 45 minutes). Lyophilization yielded a white powder, 2a (6.0 mg, 47%).

[0084] 2. AcHN-CAAAC-CONH2+9-Fluorenone(2b) The purified AcHN-CAAAC-CONH2(1b) (4.8 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 200 mM 9-fluorenone (36.0 mg, 20.0 equivalents) was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (15-60%, 45 minutes). Lyophilization yielded a white powder, 2b (2.4 mg, 38%).

[0085] 3. H2N-CYFQNCPRG-CONH2+9-Fluorenone(2c) (derived from vasopressin) JPEG2026521678000066.jpg6980H2N-CYFQNCPRG-CONH2+9-Fluorenon(2c) The purified H2N-CYFQNCPRG-CONH2(1c) (10.8 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 9-Fluorenone (90.0 mg, 50.0 equivalents) at a concentration of 200 mM was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-50%, 45 minutes). Lyophilization yielded 2c (3.4 mg, 27%) as a white powder.

[0086] 4. H2N-GGGCYFQNCPKG-CONH2+9-Fluorenone(2d) (derived from terlipressin) JPEG2026521678000067.jpg6993H2N-GGGCYFQNCPKG-CONH2+9-Fluorenon(2d) The purified H2N-GGGCYFQNCPKG-CONH2(1d) (12.3 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 200 mM 9-fluorenone (36.0 mg, 20.0 equivalents) was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-50%, 45 minutes). Lyophilization yielded a white powder of 2d (4.0 mg, 31%).

[0087] 5. AcHN-RC(D-)AH(D-)FRWC-CONH2+9-Fluorenone(2e) (derived from cetomelanotide) JPEG2026521678000068.jpg6999AcHN-RC(D-)AH(D-)FRWC-CONH2+9-Fluorenone(2e) The purified AcHN-RC(D-)AH(D-)FRWC-CONH2(1e) (11.2 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 200 mM 9-fluorenone (36.0 mg, 20.0 equivalents) was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-50%, 45 minutes). Lyophilization yielded 2e (3.8 mg, 30%) as a white powder.

[0088] 6. H2N-CYIQNCPLG-CONH2+9-Fluorenone(2f) (derived from oxytocin) JPEG2026521678000069.jpg5495H2N-CYIQNCPLG-CONH2+9-Fluorenon(2f) The purified H2N-CYIQNCPLG-CONH2(1f) (10.1 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 9-Fluorenone (90.0 mg, 50.0 equivalents) at a concentration of 200 mM was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (15-60%, 45 minutes). Lyophilization yielded 2f (4.5 mg, 38%) as a white powder.

[0089] 7. H2N-(D-)NaICY(D-)WKVCT-CONH2+9-Fluorenone (2g) (derived from lanreotide) JPEG2026521678000070.jpg6368H2N-(D-)NaICY(D-)WKVCT-CONH2+9-Fluorenone(2g) The purified H2N-(D-)NaICY(D-)WKVCT-CONH2 (1 g) (10.9 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 9-Fluorenone (90.0 mg, 50.0 equivalent) at a concentration of 200 mM was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 20% ACN / H2O and purified by preparative HPLC (20-70%, 45 minutes). Lyophilization yielded 2 g (6.6 mg, 54%) of a white powder.

[0090] 8.AcHN-GCYIQNCPLG-CONH2+cyclobutanone (2h) JPEG2026521678000071.jpg6194AcHN-GCYIQNCPLG-CONH2+Cyclobutanone (2h) The purified AcHN-GCYIQNCPLG-CONH2(1a) (11.1 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 200 mM cyclobutanone (14.0 mg, 20.0 equivalents) was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (15-60%, 45 minutes). Lyophilization yielded a white powder of 2h (5.2 mg, 45%).

[0091] 9.AcHN-GCYIQNCPLG-CONH2+Boc-4-Piperidone(2i) JPEG2026521678000072.jpg5889AcHN-GCYIQNCPLG-CONH2+Boc-4-piperidone(2i) The purified AcHN-GCYIQNCPLG-CONH2(1a) (11.1 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 200 mM N-(tert-butoxycarbonyl)-4-piperidone (39.8 mg, 20.0 equivalents) was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-50%, 45 minutes). Lyophilization yielded 2i (2.3 mg, 20%) as a white powder.

[0092] 10. AcHN-GCYIQNCPLG-CONH2+1-(prop-2-in-1-yl)piperidine-4-one(2j) JPEG2026521678000073.jpg5889AcHN-GCYIQNCPLG-CONH2+1-(prop-2-in-1-yl)piperidine-4-one(2j) The purified AcHN-CAAAC-CONH2(1a) (11.1 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 1-(prop-2-in-1-yl)piperidine-4-one (42.0 mg, 20.0 equivalents) at a concentration of 200 mM was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (15-60%, 45 minutes). Lyophilization yielded 2 joules (5.6 mg, 46%) of a white powder.

[0093] 11. AcHN-GCYIQNCPLG-CONH2+5-azidopentyl)piperidine-4-one (2k) JPEG2026521678000074.jpg5990AcHN-GCYIQNCPLG-CONH2+5-azidopentyl)piperidine-4-one(2k) The purified AcHN-GCYIQNCPLG-CONH2(1a) (11.1 mg, 1.0 equivalent) after SPPS was dissolved in TFA to a final concentration of 10 mM. 1-(5-azidopentyl)piperidine-4-one (42.0 mg, 20.0 equivalents) at a concentration of 200 mM was added to the solution, and the reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-50%, 45 minutes). Lyophilization yielded 2k (5.6 mg, 43%) white powder.

[0094] 12. AcHN-GCYIQNCPLG-CONH2+Acetone(3a) JPEG2026521678000075.jpg5986AcHN-GCYIQNCPLG-CONH2+acetone(3a) The purified AcHN-GCYIQNCPLG-CONH2(1a) (11.1 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (1.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (10-50%, 35 minutes). Lyophilization yielded a white powder 3a (6.6 mg, 63%).

[0095] 13. H2N-GGGCYFQNCPKG-CONH2 + acetone (3b) (derived from tellippressin) JPEG2026521678000076.jpg6299H2N-GGGCYFQNCPKG-CONH2+acetone(3b) The purified H2N-GGGCYFQNCPKG-CONH2(1d) (12.3 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (1.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-50%, 35 minutes). Lyophilization yielded white powder 3b (8.3 mg, 65%).

[0096] 14. AcHN-RC(D-)AH(D-)FRWC-CONH2+Acetone(3c)(derived from cetomelanotide) JPEG2026521678000077.jpg6185AcHN-RC(D-)AH(D-)FRWC-CONH2+acetone(3c) The purified AcHN-RC(D-)AH(D-)FRWC-CONH2(1e) (11.9 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (1.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (10-60%, 35 minutes). Lyophilization yielded 3c (8.9 mg, 77%), a white powder.

[0097] 15. H2N-(D-)NaICY(D-)WKVCT-CONH2+Acetone(3d)(derived from lanreotide) JPEG2026521678000078.jpg6873H2N-(D-)NaICY(D-)WKVCT-CONH2+acetone(3d) The purified H2N-(D-)NaICY(D-)WKVCT-CONH2 (1 g) (10.9 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (1.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 20% ACN / H2O and purified by preparative HPLC (20-70%, 35 minutes). Lyophilization yielded 3e (5.3 mg, 47%), a white powder.

[0098] 16. H2N-(D-)FCF(D-)WKTCT-CH2OH + Acetone(3e) (derived from octreotide) JPEG2026521678000079.jpg6268H2N-(D-)FCF(D-)WKTCT-CH2OH+Acetone(3e) The purified H2N-(D-)FCF(D-)WKTCT-CH2OH(1h) (10.6 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (1.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 15% ACN / H2O and purified by preparative HPLC (15-60%, 35 minutes). Lyophilization yielded 3e (7.5 mg, 71%), a white powder.

[0099] 17. H2N-RLCRIVVIRVCR-COOH + Acetone (3f) (derived from bactennessin) JPEG2026521678000080.jpg69126H2N-RLCRIVVIRVCR-COOH+acetone(3f) The purified H2N-RLCRIVVIRVCR-COOH(1i) (7.4 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (10.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 0.5 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 10% ACN / H2O and purified by preparative HPLC (10-60%, 35 minutes). Lyophilization yielded 3f (1.1 mg, 15%), a white powder.

[0100] 18. H2N-AGCKNFFWKTFTSC-CONH2 + Acetone (3g) (derived from somatostatin) JPEG2026521678000081.jpg8884H2N-AGCKNFFWKTFTSC-CONH2+Acetone(3g) The purified H2N-AGCKNFFWKTFTSC-CONH2(1j) (16.4 mg, 1.0 equivalent) after SPPS was dissolved in a mixed solvent (1.0 mL) of TFA and acetone (v:v=1:1) to a final concentration of 10 mM. The reaction mixture was stirred at room temperature for approximately 8 hours. After the reaction, the solvent was blown off under a stream of N2, and the crude peptide product was subsequently precipitated with diethyl ether. The residue was dissolved in 10 mL of 20% ACN / H2O and purified by preparative HPLC (20-70%, 35 minutes). Lyophilization yielded 3 g (8.6 mg, 51%) of a white powder.

[0101] It should be understood that the disclosed methods and compositions are not limited to the specific methods, solutions, and reagents described, for this reason being that they may vary. It should also be understood that the terms used herein are used solely to describe specific embodiments and are not intended to limit the scope of the invention, and the scope of the invention is limited by the appended claims.

[0102] Those skilled in the art will be able to recognize or identify many equivalents to the methods and specific embodiments of the compositions described herein by performing ordinary experiments. These equivalents are intended to be covered by the following claims.

Claims

1. The reaction mixture is maintained at a temperature sufficient to form the product for a sufficient amount of time. The reaction mixture comprises a peptide containing two or more cysteine ​​residues, a cyclic ketone reagent or acetone, and a solvent. The product comprises a thioacetalized peptide, and the thioacetalization binds two cysteine ​​residues of the peptide. A method for late-stage cyclization of peptides.

2. The method according to claim 1, wherein the peptide is linear or cyclic (including monocyclic, bicyclic, etc.).

3. The method according to claim 1 or 2, wherein the peptide is a random peptide or a peptide pharmaceutical.

4. The method according to any one of claims 1 to 3, wherein the peptide is formed from natural amino acids.

5. The method according to any one of claims 1 to 4, wherein the cyclic ketone reagent is a cyclic ketone reagent having the following structure. Equation I (In the formula, R and R' are independently alkyl groups, and Either R and R' do not exist, or R and R' together form annular portion A. Here, A is a cycloalkyl group, a cycloalkenyl group, a cycloalkynyl group, an aryl group, a polyaryl group, a heteroaryl group, a heteropolyaryl group, or a heterocyclic group. Here, R'' represents a hydrogen atom or a substituent on the cyclic portion A, and each time R'' appears, it independently represents a benzyl group, an allyl group, an alkyl group (e.g., C1-C6 alkyl groups), a halogen, -CN, or -CF. 3、 - NO 2 , an alkoxy group or an aryl group, N is an integer between 0 and 10, 0 and 8, 0 and 6, 0 and 4, or 0 and 2.

6. The method according to claim 5, wherein R and R' are independently alkyl groups such as methyl groups.

7. The method according to claim 5 or 6, wherein A is cyclobutane, azetidine, cyclopentane, fluorenone, cyclohexane, or piperidine.

8. The method according to any one of claims 1 to 7, wherein the solvent is trifluoroacetic acid.

9. The method according to claim 8, wherein trifluoroacetic acid acts as a catalyst and is the only solvent in the reaction mixture.

10. The method according to any one of claims 1 to 9, wherein the reaction mixture is maintained at a temperature of 20°C to 35°C, such as about 30°C, for a maximum of 1 hour, a maximum of 2 hours, a maximum of 3 hours, 10 minutes to 1 hour, 20 minutes to 2 hours, or 30 minutes to 3 hours.

11. The method according to any one of claims 1 to 10, wherein the molar ratio of the peptide to the cyclic ketone reagent (peptide:cyclic ketone reagent) is 0.1 to 1, for example, about 0.

2.

12. The method according to any one of claims 1 to 11, wherein the cyclic ketone reagent has any of the following structures. (In the formula, R 1 and R 3 is H, benzyl group, allyl group, alkyl group (e.g., C1-C6 alkyl group), and Here, each R 2 These are independently H, benzyl group, allyl group, alkyl group (e.g., C1-C6 alkyl group), halogen, -CN, -CF 3 , -NO 2 (It is an alkoxy group or an aryl group.)

13. The method according to any one of claims 1 to 12, wherein the thioacetalized peptide has any of the following structures. 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , or 。

Images