Synthesis of amanine and its derivatives.

Abstract

The present invention relates to the chemical synthesis of amanine and its derivatives. The present invention also relates to intermediates in the synthesis of amanine.

Description

[Technical field]

[0001] The present invention relates to the chemical synthesis of flaxseed and its derivatives. This relates to intermediate products in the synthesis of nin. [Background technology]

[0002] 2. Background of the Invention The object of the present invention is to provide means and methods for the chemical synthesis of amanine or its derivatives. This object is achieved by the subject matter of the independent claims herein. Summary of the Invention

[0003] Summary of the Invention A first aspect of the present invention relates to a process for the preparation of a compound of formula (I). [ka] Another aspect relates to intermediates in the synthesis of amanine. [Brief description of the drawings]

[0004] [Figure 1] Synthesis of the dipeptide building block H-Asp(OAll)-Hyp-OFm°HCl (3) [Diagram 2] Synthesis of deoxyamannin DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0005] Detailed Description of the Invention Terms and Definitions The amino acid sequences are written from the amino terminus to the carboxyl terminus. Capital letters in sequence positions refer to L-amino acids in one-letter code (Stryer, Biochemistry, 3rd ed. p. 21). Lower case letters at amino acid sequence positions refer to the corresponding D- or (2R)-amino acid.

[0006] The term AA in this context relates to an amino acid.

[0007] The term "protecting group" in the context of this specification relates to a moiety covalently attached to a functional group (particularly a carboxylic acid, amino or hydroxyl moiety of the molecules discussed herein) that can be selectively attached to and removed from the functional group without affecting the integrity or chiral orientation of the carbon backbone of the molecule or cleaving certain other protecting groups attached to the molecule.

[0008] The term "deprotecting agent" in the present context relates to an agent capable of cleaving a particular protecting group. Those skilled in the art can select a deprotecting agent according to the protecting group. The condition under which the protecting group is cleavable constitutes the deprotecting agent, for example, if the protecting group is cleavable under acidic conditions, the deprotecting agent is an acid.

[0009] The term "preactivated carboxyl group" in the present context relates to a carboxyl moiety which reacts to an activated ester susceptible to nucleophilic attack of an amine group to form a peptide bond.

[0010] The term "preactivated amino group" in the present context relates to an amino group which reacts with increased nucleophilicity to an N-trimethylsilylamine to attack a carboxylic acid moiety to form a peptide bond.

[0011] A comprehensive review of modern protecting group chemistry, particularly as it relates to the compounds disclosed herein, is available in Peter GM Wuts, Greene's Protective Groups in Organic Synthesis, 5th Edition, Wiley 2014.

[0012] US6693178B2 - "Protecting groups useful in the synthesis of polysaccharides, natural products, and combinatorial libraries" and US20160024143A1 - "Deprotection methods" are incorporated herein by reference.

[0013] In this specification, we follow the standard convention of organic chemistry that unspecified positions in a formula are assumed to be saturated carbons.

[0014] A first aspect of the present invention relates to a process for the preparation of a compound of formula (I),

[0015] [ka] where a) in reaction step a), a compound of formula (IIa)

[0016] [ka] with a peptide bond forming reagent, specifically a coupling reagent selected from a carbodiimide, an imidazolinium reagent, a phosphonium salt, an organophosphorus reagent, an uronium salt, a pyridinium reagent, and a phosphonic acid; More specifically, reacting with HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU, If applicable, the compound may be R PGP and / or R PGOH and / or R AL and / or R AMIN reacting with a deprotecting agent which removes

[0017] or here b) In reaction step b), a compound of formula (IIb) [ka] a peptide bond forming reagent, Specifically, coupling reagents selected from carbodiimides, imidazolinium reagents, phosphonium salts, organophosphorus reagents, uronium salts, pyridinium reagents, and phosphonic acids; More specifically, reacting with HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU, If applicable, the compound may be R PGP and / or R PGOH and / or R AL and / or R AMIN reacting with a deprotecting agent which removes

[0018] or here c) In reaction step c), a compound of formula (IIc) [ka] with a peptide bond forming reagent, specifically a coupling reagent selected from a carbodiimide, an imidazolinium reagent, a phosphonium salt, an organophosphorus reagent, an uronium salt, a pyridinium reagent, and a phosphonic acid; More specifically, reacting with HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU,

[0019] If applicable, the compound may be R PGP and / or R PGOH and / or R AL and / or R AMIN reacting with a deprotecting agent which removes

[0020] or here d) In reaction step d), a compound of formula (IId) [ka] a peptide bond forming reagent, Specifically, coupling reagents selected from carbodiimides, imidazolinium reagents, phosphonium salts, organophosphorus reagents, uronium salts, pyridinium reagents, and phosphonic acids; More specifically, reacting with HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU, If applicable, the compound may be R PGP and / or R PGOH and / or R AL and / or R AMIN reacting with a deprotecting agent which removes

[0021] During the ceremony X 1 , X 2 , X 3 , and X 4 , and Y 1 , Y 2 , Y 3 , and Y 4 is H, or Y 1 , Y 2 , Y 3 , and Y 4 one, two, three or all of Y are OH or NH2 and all other Y are H and the corresponding X is NHR AMIN OR PGP and all other X's are H, Here, R PGP is a protecting group for a phenolic OH group, specifically a phenolic OH protecting group that is not acid or alkali labile, more specifically a protecting group that is cleavable under reducing conditions, most specifically Cbz; Or, R AMIN is a protecting group for a phenolic amino group, specifically a phenolic amino protecting group that is not acid or alkali labile, more specifically a protecting group that is cleavable under reductive conditions, most specifically Cbz, or Y 1 , Y 2 , Y 3 , and Y 4 and the corresponding X is F, Cl, Br, I, CN, NO2, acyl, N3, or alkyne, and all other X and Y are H; Specifically, X and Y are H, or Y 1 , Y 2 , Y3 , and Y 4 One of the groups is OH and the corresponding X is OR PGP and all other X and Y are H; Z and W are H, or Z is OH and W is OR PGOH where R PGOH is a protecting group for a hydroxyl group, specifically, a protecting group for a hydroxyl group that can be cleaved by a fluoride ion, R AL is a protected carboxyl group or a 1-5AA peptide, specifically, R AL is an O-allyl or O-methyl ester or a 1-5AA peptide, more specifically, R AL is an O-allyl or O-methyl ester, most specifically R AL is O-allyl; V is OH or a 1-5AA peptide, specifically, V is OH; Q is S or SO.

[0022] In certain embodiments, the method is carried out via step a) or step b). In certain embodiments, the 1-5AA peptide is composed of proteinogenic amino acids.

[0023] In certain embodiments, a compound of formula (III) [ka] with a compound of formula (IV),

[0024] [ka] During the ceremony X 1 , X 2 , X 3 , X 4 , W, Q, and R AL has the same meaning as defined above; R COOYis a carboxyl protecting group, specifically fluorenylmethyl or benzyl, more specifically fluorenylmethyl; Where: (III) and (IV) are reacted in reaction step (e) with a peptide bond forming reagent, specifically HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU, to obtain a compound characterized by (IIa).

[0025] In certain embodiments, a compound of formula (V) [ka] with a compound of formula (VI),

[0026] [ka]

[0027] During the ceremony, X 1 , X 2 , X 3 , X 4 , Q, and W have the same meaning as defined above; R NHB is an amino protecting group, specifically an amino protecting group that is cleavable under alkaline conditions, more specifically Fmoc; Here, in the reaction step (f), The amino group of (V) is preactivated, particularly with MSA, and the preactivated (V) and (VI) are reacted with a peptide bond forming reagent, particularly HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU, more particularly COMU, to obtain compound (III).

[0028] In certain embodiments, a compound of formula (V) [ka] with a compound of formula (VII),

[0029] [ka]

[0030] During the ceremony, X 1 , X 2 , X 3 , X 4 , W, Q, R NHB , and R AL has the same meaning as defined above; Where: In the reaction step (g), preactivating the amino group of (V), in particular with MSA, and reacting the preactivated (V) and (VII) with a peptide bond forming reagent, in particular HATU; or The amino group of (V) is preactivated, in particular with MSA, the carboxyl group of compound (VII) is preactivated, in particular with O-PFP-ester, O-PCP-ester, or OSu-ester, and the preactivated (V) and the preactivated (VII) are reacted to give compound (IIb).

[0031] When coupling compounds (V) and (VII), it is not necessary to protect the acid COOH group of compound (V). Even without a protecting group, no significant side reactions were observed.

[0032] In certain embodiments, a compound of formula (V) [ka]

[0033] with a compound of formula (VII), [ka]

[0034] During the ceremony, X 1 , X 2 , X 3 , X 4 , W, Q, R NHB, and R AL has the same meaning as defined above; Here, (V) and (VII) are reacted with a peptide bond forming reagent, specifically HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU, to give compound (IIc).

[0035] In certain embodiments, a compound of formula (XVI) [ka] with a compound of formula (XVII),

[0036] [ka] During the ceremony, X 1 , X 2 , X 3 , X 4 , W, Q, and R AL has the same meaning as defined above; R NHB2 is an amino protecting group, specifically an amino protecting group that is cleavable under acidic conditions, more specifically Boc; Where: preactivating the amino group of (XVI), in particular with MSA, and reacting the preactivated (XVI) and (XVII) with a peptide bond forming reagent, in particular HATU; or The amino group of (XVI) is preactivated, in particular with MSA, the carboxyl group of compound (XVII) is preactivated, in particular with O-PFP-ester, O-PCP-ester, or OSu-ester, and the preactivated (XVI) and the preactivated (XVII) are reacted to give compound (IId).

[0037] In certain embodiments, a compound of formula (V) [ka] with a compound of formula (XVIII),

[0038] [ka]

[0039] During the ceremony, X 1 , X 2 , X 3 , X 4 , W, Q, R NHB , and R AL has the same meaning as defined above; Where: preactivating the amino group of (V), in particular with MSA, and reacting the preactivated (V) and (XVIII) with a peptide bond forming reagent, in particular HATU; or The amino group of (V) is preactivated, in particular with MSA, the carboxyl group of compound (XVIII) is preactivated, in particular with O-PFP-ester, O-PCP-ester, or OSu-ester, and the preactivated (V) and preactivated (XVIII) are reacted to give compound (XVI).

[0040] In certain embodiments, for compounds of formula (Idesox), the sulfur atom is oxidized. [ka] In the formula, Y 1 , Y 2 , Y 3 , Y 4 , Z, Q, and V have the same meanings as defined above.

[0041] In certain embodiments, the oxidation of the sulfur atom is carried out using manganese ions. In certain embodiments, the compound is reacted with a compound of formula (XV), Mn(OTf)2 and H2O2. [ka]

[0042] In certain embodiments, the oxidation of sulfur atoms is carried out using PPO, dibenzoyl peroxide, tert-butyl perbenzoate, or lauroyl peroxide.The preparation of PPO is described in S. Gan, J. Yin, Y. Yao, Y. Liu, D. Chang, D. Zhu, L. Shi, Org. Biomol. Chem. 2017, 15, 2647-2654.

[0043] In a particular embodiment, the oxidation of the sulfur atom is carried out with mCPBA (metachloroperbenzoic acid) in isopropanol / ethanol (8:3).

[0044] In certain embodiments, oxidation of sulfur atoms is carried out with oxaziridinium salts as described in Rio et al, Org. Lett. 2007, 9,12, 2265-2268.

[0045] In certain embodiments, the oxidation of the sulfur atom is carried out with a non-enantioselective agent, or simply with oxygen or hydrogen peroxide. In certain embodiments, the oxidation of sulfur atoms is carried out using iodine and oxygen.

[0046] In reaction step (h), the sulfur atom is oxidized to obtain compound (Iox). [ka]

[0047] In certain embodiments, for compounds of formula (Vdesox), the sulfur atom is oxidized. [ka] In the formula, X 1 , X 2 , X 3 , and X 4 has the same meaning as defined above.

[0048] In certain embodiments, the oxidation of the sulfur atom is carried out using manganese ions. In certain embodiments, the compound is reacted with a compound of formula (XV), Mn(OTf)2, and H2O2. [ka]

[0049] In certain embodiments, the oxidation of sulfur atoms is carried out using PPO, dibenzoyl peroxide, tert-butyl perbenzoate, or lauroyl peroxide.The preparation of PPO is described in S. Gan, J. Yin, Y. Yao, Y. Liu, D. Chang, D. Zhu, L. Shi, Org. Biomol. Chem. 2017, 15, 2647-2654.

[0050] In a particular embodiment, the oxidation of the sulfur atom is carried out with mCPBA (metachloroperbenzoic acid) in isopropanol / ethanol (8:3).

[0051] In certain embodiments, oxidation of sulfur atoms is carried out with oxaziridinium salts as described in Rio et al, Org. Lett. 2007, 9,12, 2265-2268.

[0052] In certain embodiments, the oxidation of the sulfur atom is carried out with a non-enantioselective agent, or simply with oxygen or hydrogen peroxide.

[0053] In certain embodiments, the oxidation of sulfur atoms is carried out using iodine and oxygen.

[0054] In reaction step (i), the sulfur atom is oxidized to obtain a compound (Vox). [ka]

[0055] In certain embodiments, a compound of formula (VIII) [ka]

[0056] (In the formula, R NHF is an amino protecting group, specifically an amino protecting group cleavable by fluoride ion or strong acid, more specifically Teoc, or an amino protecting group cleavable under alkaline conditions, more specifically Fmoc; R COOA is a carboxyl protecting group, specifically a carboxyl protecting group that is cleavable under strongly acidic conditions, more specifically tert-butyl; X 1 , X 2 , X 3 , and X 4 has the same meaning as explained above.) in reaction step (j) with a peptide bond forming reagent, specifically a coupling reagent selected from carbodiimides, imidazolinium reagents, phosphonium salts, organophosphorus reagents, uronium salts, pyridinium reagents, and phosphonic acids, more specifically T3P, HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU; This compound is reacted with R NHF and R COOA Reaction with a deprotecting agent, particularly TFA, to remove the protective group gives compounds characterized as (Vdesox) or (V).

[0057] In certain embodiments, a compound of formula (IX) [ka] with a compound of formula (X),

[0058] [ka]

[0059] During the ceremony, R NHAis an amino protecting group, specifically an amino protecting group that is cleavable under acidic conditions, more specifically Boc; R COOA , R NHF And X 1 , X 2 , X 3 , and X 4 has the same meaning as explained above, Where: In the reaction step (l), Compound (VII) is preactivated with a peptide bond forming reagent, in particular HATU, COMU, HBTU, TBTU, TOMBU, COMBU or HCTU, and then reacted with silylated compound (VIII); or Pre-activation in OSu-ester followed by reaction with compound (VII),

[0060] In reaction step (m), the above compound is reacted with R NHA with a deprotecting agent which removes the A compound characterized by the formula (VIII) is obtained.

[0061] In certain embodiments, a compound of formula (XI) [ka] with a compound of formula (XII),

[0062] [ka]

[0063] During the ceremony, R COOZ is a carboxyl protecting group, specifically a Zn-cleavable carboxyl protecting group, more specifically Tce, or R COOZ is H, R COOA , R NHF , RNHA And X 1 , X 2 , X 3 , and X 4 has the same meaning as explained above, In the reaction step (n), reacting R COOZ is a carboxyl protecting group, in reaction step (o), the compound is COOZ with a deprotecting agent, particularly Zn, which removes A compound characterized by (IX) is obtained.

[0064] A protecting group strategy that relies on acid stability was applied. The deprotection was carried out by lowering the pH. First, the Tce group of tryptophan (R COOZ ) was removed under reductive conditions with Zn at slightly acidic pH. Then, the cysteine ​​(R of compound IX) was removed with p-toluenesulfonic acid. NHA Finally, the Teoc group of compound (V) was removed. NHF ) and tert-butyl (R COOA ) was simultaneously removed using 95% TFA.

[0065] In certain embodiments, a compound of formula (VI) [ka] with a compound of formula (XIII),

[0066] [ka] and reacting with a compound of formula (XIV)

[0067] [ka]

[0068] During the ceremony R AL has the same meaning as explained above, R Pepis an activated ester, in particular an O-pentafluorophenol or an OSu-ester, R NHB is an amino protecting group, specifically an amino protecting group that is cleavable under alkaline conditions, more specifically Fmoc; In the reaction step (p), the reaction is carried out by solid phase peptide synthesis, in which the carboxyl group of compound (XIV) may be protected, A compound characterized by the formula (VII) is obtained.

[0069] Another aspect of the present invention relates to compounds of general formula (I): [ka]

[0070] During the ceremony, Y 1 , Y 2 , and Y 4 one, two or three of Y are OH and all other Y are H; or Y 1 , Y 2 , Y 3 , and Y 4 one, two, three or all are selected from F, Cl, Br, and I, and all other Y's are H; Specifically, Y 1 , Y 2 , and Y 4 one of Y is OH and all other Y are H; Z is H or Z is OH; V is OH or a 1-5AA peptide, in particular V is OH; Q is S or SO.

[0071] Another aspect of the present invention relates to compounds of general formula (IIa): [ka]

[0072] During the ceremony X1 , X 2 , X 3 , and X 4 is H, or X 1 , X 2 , X 3 , and X 4 one, two, three or all of X are OH and all other X are H; or X 1 , X 2 , X 3 , and X 4 one, two, three or all of X are selected from F, Cl, Br, and I, and all other X are H; Specifically, X 1 , X 2 , X 3 , and X 4 is H or X 1 , X 2 , X 3 , and X 4 one of X is OH and all other X are H; W is H or W is OH; V is OH or a 1-5AA peptide, in particular V is OH; Q is S or SO.

[0073] Another aspect of the present invention relates to compounds of general formula (IIb): [ka]

[0074] During the ceremony, X 1 , X 2 , X 3 , and X 4 is H, or X 1 , X 2 , X 3 , and X 4 one, two, three or all of X are OH and all other X are H; or X 1 , X 2, X 3 , and X 4 one, two, three or all of X are selected from F, Cl, Br, and I, and all other X are H; Specifically, X 1 , X 2 , X 3 , and X 4 is H or X 1 , X 2 , X 3 , and X 4 one of X is OH and all other X are H; W is H or W is OH; V is OH or a 1-5AA peptide, in particular V is OH; Q is S or SO.

[0075] Another aspect of the present invention relates to compounds of general formula (III): [ka]

[0076] During the ceremony, X 1 , X 2 , and X 4 one, two or all of X are OH and all other X are H; or X 1 , X 2 , and X 4 one, two or all of X are selected from F, Cl, Br, and I, and all other X are H; Specifically, X 1 , X 2 , and X 4 one of X is OH and all other X are H; W is H or W is OH; Q is S or SO.

[0077] Another aspect of the present invention relates to compounds of general formula (V): [ka] During the ceremony, X 1 , X 2 , and X 4 one, two or all of X are OH and all other X are H; or X 1 , X 2 , and X 4 one, two or all of X are selected from F, Cl, Br, and I, and all other X are H; Specifically, X 1 , X 2 , and X 4 one of X's is OH and all other X's are H; Q is S or SO.

[0078] The present invention is further illustrated by the following examples and figures from which further embodiments and advantages can be derived, these examples are intended to illustrate the invention and are not intended to limit the scope of the invention.

[0079] Text description of the illustration image024.gif. Figure 1: Synthesis of the dipeptide building block H-Asp(OAll)-Hyp-OFm°HCl (3) Figure 2: Synthesis of deoxyamannin EXAMPLES

[0080] Synthesis of (N-Boc)2-Cystine-(OtBu)2 (35) [ka]

[0081] A solution of L-cystine-(OtBu)2 (34, 10 g, 24 mmol, 1.0 equiv.) in a 1:1 mixture of H2O / dioxane (240 mL) was treated with NaHCO3 (8.06 g, 96.0 mmol, 4.00 equiv.) and Boc2O (10.1 mL, 47.0 mmol, 2.00 equiv.) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure and the aqueous layer was extracted with EtOAc (3 x 120 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4, and the solvent was removed under reduced pressure to give 35 (13.2 g, 24.0 mmol, quantitative) as a pale yellow solid. HRMS(ESI):m / z calculated value C 24 H 44 N2O8S2(M+H) + 553.2612,Measurement value 553.2615.

[0082] tert-Butyl S-(6-(benzyloxy)-3-((S)-3-oxo-3-(2,2,2-trichloroethoxy)-2-(((2-(trimethylsilyl)ethoxy)carbonyl)amino)propyl)-1H-indol-2-yl)-N-(tert-butoxycarbonyl)-L-cysteinate (40) [ka]

[0083] To a solution of (N-Boc)2-L-cystine-(OtBu)2 (35,900 mg, 1.63 mmol, 1.00 equiv.) in CHCl3 (16.3 mL) was added SO2Cl2 (263 μL, 3.26 mmol, 2.00 equiv.). The reaction mixture was stirred at room temperature for 1 h, after which the solvent was removed under reduced pressure. The residue was dissolved in CHCl3 (16.3 mL), cooled to 0 °C, and added to an ice-cold solution of 39 (800 mg, 1.67 mmol, 1.00 equiv.) and NaHCO3 (420 mg, 5.00 mmol, 3.00 equiv.) in CHCl3 (16.7 mL) was added dropwise over 10 min. The reaction mixture was then stirred at 0 °C for 15 min and at room temperature for 1 h. The organic layer was washed with H2O (10 mL) and saturated NaHCO3 solution (10 mL). After the organic layer was dried over Na2SO4 and the solvent was removed under reduced pressure, the crude product 40 was used in the next step without further purification. HRMS(ESI):m / z calculated value C 38 H 52 Cl3N3O9SSi (M+H) + 860.2332,Measured value 860.2323.

[0084] tert-Butyl N-(tert-butoxycarbonyl)-S-(3-((S)-3-oxo-3-(2,2,2-trichloroethoxy)-2-(((2-(trimethylsilyl)ethoxy)carbonyl)amino)propyl)-1H-indol-2-yl)-L-cysteinate (41) [ka] To a solution of (N-Boc)2-L-cystine-(OtBu)2 (39, 5.06 g, 9.15 mmol, 1.00 equiv.) in CHCl3 (92 mL) was added SO2Cl2 (1.48 mL, 18.3 mmol, 2.00 equiv.). The reaction mixture was stirred at room temperature for 1 h, after which the solvent was removed under reduced pressure. The residue was dissolved in CHCl3 (92 mL), cooled to 0 °C, and added to an ice-cold solution of 38 (4.4 g, 9.17 mmol, 1.00 equiv.) and NaHCO3 (2.31 g, 27.5 mmol, 3.00 equiv.) in CHCl3 (92 mL) was added dropwise over 10 min. The reaction mixture was then stirred at 0 °C for 15 min and at room temperature for 1 h. The organic layer was washed with H2O (2 × 100 mL) and saturated NaHCO3 solution (2 × 80 mL). After the organic layer was dried over Na2SO4 and the solvent was removed under reduced pressure, the crude product 41 was used in the next step without further purification. HRMS(ESI):m / z calculated value C 31 H 46 Cl3N3O8SSi (M+H) + 754.1913,Measured value 754.1917.

[0085] (S)-3-(6-(benzyloxy)-2-(((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropyl)thio)-1H-indol-3-yl)-2-(((2-(trimethylsilyl)ethoxy)carbonyl)amino)propionic acid (49): [ka]

[0086] A solution of tryptathionine derivative 39 (1.63 mmol, 1.00 equiv.) in DMF (8.4 mL) was treated with CH3COOH (0.8 mL) and zinc (3.51 g, 53.6 mmol, 33.0 equiv.) for 2 h at 45 °C. The reaction mixture was filtered over Celite and the solvent was removed under reduced pressure. The crude product was dissolved in EtOAc (50 mL) and washed with 10% KHSO4 solution (2 × 25 mL) and brine (2 × 25 mL). After drying over Na2SO4 and removing the solvent under reduced pressure, the crude product was purified by C18 reverse phase chromatography (AcN / H2O 50% to 100% gradient) to give compound 49 as a yellow solid (840 mg, 83% for two steps). HRMS(ESI):m / z calculated value C 36 H 51 N3O9SSi (M+H) + 730.3183,Measured value 730.3188.

[0087] (S)-3-(2-(((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropyl)thio)-1H-indol-3-yl)-2-(((2-(trimethylsilyl)ethoxy)carbonyl)amino)propionic acid (50) [ka]

[0088] A solution of tryptathionine derivative 38 (9.15 mmol, 1.00 equiv.) in DMF (40 mL) was treated with CH3COOH (4 mL) and zinc (20.0 g, 302 mmol, 33.0 equiv.) for 2 h at 45 °C. The reaction mixture was filtered over Celite and the solvent was removed under reduced pressure. The crude product was dissolved in EtOAc (200 mL) and washed with 10% KHSO4 solution (2 × 50 mL) and brine (2 × 50 mL). After drying over Na2SO4 and removal of the solvent under reduced pressure, the crude product was purified by C18 reverse phase chromatography (AcN / H2O 50% to 100% gradient) to give compound 50 as a yellow oil (5.0 g, 88% for two steps). HRMS(ESI):m / z calculated value C 31 H46 Cl3N3O8SSi(M+H) + 624.2769,Measured value 624.2775.

[0089] ((Benzyloxy)carbonyl)glycyl-L-isoleucine (44) [ka]

[0090] To a solution of Cbz-glycine (42, 10.0 g, 32.7 mmol, 1.00 equiv.) in acetone (100 mL) was added a suspension of L-isoleucine (4.71 g, 35.9 mmol, 1.10 equiv.) and NaHCO3 (8.23 g, 87.9 mmol, 3.00 equiv.) in water (100 mL). The reaction mixture was stirred at room temperature for 3 h and concentrated under reduced pressure. The aqueous layer was carefully acidified to pH = 4 by dropwise addition of 1 M HCl and extracted with EtOAc (3 x 150 mL). The organic layer was then washed with brine (2 x 100 mL), dried over Na2SO4, and evaporated under reduced pressure to give the product 44 as a colorless oil (10.1 g, 96%). HRMS(ESI):m / z calculated value C 16 H 22 N2O5(M+H) + 323.1601, measured value 323.1606

[0091] Glycyl-L-isoleucylglycine (45): [ka]

[0092] Dipeptide 44 (10.1 g, 31.3 mmol, 1.00 equiv.) and benzyl glycinate (8.21 g, 40.7 mmol, 1.30 equiv.) were dissolved in dry DMF (125 mL). Then, COMU (17.4 g, 40.7 mmol, 1.30 equiv.) and DIPEA (12.6 mL, 72.1 mmol, 3.00 equiv.) were added at 0° C. The reaction mixture was left to reach room temperature overnight and then diluted with EtOAc (300 mL). After washing with 10% KHSO4-solution (2×100 mL), the fully protected tripeptide precipitated in the organic layer. The organic layer was cooled to 4° C. for 4 h to precipitate the peptide, which was filtered and washed with cold EtOAc. The precipitate was redissolved in a 1:1 mixture of water and THF (260 mL). Pd / C (1 g) was added to the solution after degassing with N2 for 30 min. The reaction mixture was then degassed with hydrogen for 1 h. After vigorously stirring at room temperature under 1 atm of hydrogen overnight, the catalyst was filtered through a pad of Celite. The mixture was then concentrated under reduced pressure to give the product 45 as a white solid (5.71 g, 74%). HRMS(ESI):m / z calculated value C 10 H 19 N3O4(M+H) + 246.1448,Measured value 246.1440.

[0093] Synthesis of pentapeptide 51: [ka]

[0094] A solution of thioether building block 49 (111 mg, 0.14 mmol, 1.00 equiv) in AcN (0.7 mL) was treated with collidine (37 μL, 0.27 mmol, 2.0 equiv) and N,N'-disuccinimidyl carbonate (39 mg, 0.15 mmol, 1.1 equiv) and stirred at room temperature for 1 h. A solution of tripeptide 45 (44 mg, 0.18 mmol, 1.3 equiv) in a 1:4 mixture of AcN / H2O (1 mL) was added and the reaction mixture was stirred at room temperature for 2 h. The mixture was then diluted with EtOAc (20 mL), 10% KHSO4 solution (20 mL) was added and the aqueous layer was extracted with EtOAc (2 × 20 mL). The organic layer was washed with brine (2 × 0 mL), dried over Na2SO4 and evaporated in vacuo to give pentapeptide 51 as a yellow solid (115 mg, 90%). HRMS(ESI):m / z calculated value C 46 H 68 NO 12 SSi (M+H) + 957.4458,Measured value 957.4457.

[0095] Synthesis of pentapeptide 52: [ka]

[0096] A solution of tryptathionine building block 50 (2.0 g, 2.5 mmol, 1.0 equiv) in AcN (10 mL) was treated with collidine (659 μL, 4.95 mmol, 2.00 equiv) and N,N'-disuccinimidyl carbonate (697 mg, 2.72 mmol, 1.10 equiv) and stirred at room temperature for 1 h. A solution of tripeptide 45 (790 mg, 3.22 mmol, 1.30 equiv) in a 1:4 mixture of AcN / H2O (18 mL) was added and the reaction mixture was stirred at room temperature for 2 h. The mixture was then diluted with EtOAc (100 mL), 10% KHSO4 solution (20 mL) was added, and the aqueous layer was extracted with EtOAc (2 x 50 mL). The organic layer was washed with brine (2×50 mL), dried over Na 2 SO 4 and evaporated in vacuo to give pentapeptide 52 as a yellow solid (2.15 g, 93%). HRMS(ESI): m / z calculated value (M+H) + C 36 H 51 Cl3N6O 11 S851.4039, measured value 851.4058.

[0097] Fully protected cyclic pentapeptide 53: [ka]

[0098] Pentapeptide 51 (151 mg, 0.180 mmol, 1.00 equiv) was dissolved in 1 mL of p-toluenesulfonic acid in THF (1.8 M) and stirred at room temperature for 4 h. The reaction mixture was then neutralized by addition of DIPEA (320 μL, 1.84 mmol, 10 equiv) and diluted with DCM (180 mL). DIPEA (60.2 μL, 354 μmol, 2.00 equiv) and T3P (50% in EtOAc, 210 μL, 354 μmol, 0.34 equiv) were then added. The solution was stirred at room temperature for 16 h, after which 2 / 3 of the solvent was concentrated under reduced pressure. The organic layer was washed with 10% KHSO4-solution (20 mL), saturated NaHCO3-solution (20 mL), water (20 mL) and brine (20 mL), dried over Na2SO4 and the solvent was removed under reduced pressure. The crude product was purified by C18 reverse-phase chromatography (AcN / HO 50% to 100% gradient) to give the cyclic pentapeptide 53 as a yellow solid (82 mg, 70%). HRMS(ESI):m / z calculated value C 41 H 58 N6O9SSi (M+H) + 839.3828,Measured value 839.3839.

[0099] Fully protected cyclic pentapeptide (54): [ka]

[0100] Pentapeptide 52 (700 mg, 0.822 mmol, 1.00 equiv) was dissolved in 10 mL of 2M HCl in dioxane and stirred at room temperature for 40 min. The reaction mixture was then diluted with 40 mL of dioxane and the solvent was removed under reduced pressure. The precipitate was dissolved in 8 mL DMF and diluted with 82 mL DCM. DIPEA (279 μL, 1.64 mmol, 2.00 equiv) and T3P (50% in EtOAc, 977 μL, 1.64 mmol, 2.00 equiv) were then added. The solution was stirred at room temperature for 5 h, after which 1 / 3 of the solvent was concentrated under reduced pressure. The organic layer was washed with 10% KHSO4-solution (20 mL), saturated NaHCO3-solution (20 mL), water (20 mL) and brine (20 mL). The organic layer was dried over Na2SO4 and the solvent was removed under reduced pressure. The crude product was purified by C18 reverse-phase chromatography (AcN / HO 50% to 100% gradient) to give the cyclic pentapeptide 54 as a yellow solid (420 mg, 72%). HRMS(ESI):m / z calculated value C 34 H 52 N6O8SSi (M+H) + 733.3409,Measurement value 733.3409.

[0101] Fully deprotected monocyclic pentapeptide 55: [ka]

[0102] Monocyclic pentapeptide 53 (125 mg, 0.17 mmol, 1.00 equiv.) was stirred in TFA / DCM / TIPS (8:1.5:0.5) for 2 h at room temperature. The solvent was removed under reduced pressure and the crude product was purified by C18 reverse phase chromatography (AcN / HO 20%-100%) to give the fully deprotected monocyclic pentapeptide 55 as a white powder (100 mg, 100 mL). HRMS(ESI):m / z calculated value C 31 H 38 N6O7S (M+H) + 639.2595,Measured value 639.2590.

[0103] Fully deprotected monocyclic pentapeptide 56: [ka]

[0104] Monocyclic pentapeptide 54 (250 mg, 0.34 mmol, 1.00 equiv.) was stirred in TFA / DCM / TIPS (8:1.5:0.5) for 2 h at room temperature. The solvent was removed under reduced pressure and the crude product was purified by C18 reverse phase chromatography (AcN / HO 10%-30%) to give the fully deprotected monocyclic pentapeptide 56 as a white powder (200 mg, quantitative). HRMS(ESI):m / z calculated value C 24 H 32 N6O6S (M+H) + 533.2177,Measured value 533.2188.

[0105] Monocyclic hexapeptide 67: [ka]

[0106] A solution of the fully deprotected monocyclic pentapeptide 55 (42 mg, 0.66 mmol, 1.00 equiv.) and MSA (11.6 μL, 0.723 mmol, 1.10 equiv.) in DMA (2 mL) was stirred for 2 h at 50 °C. At the same time, a solution of Fmoc-DHIle(TBS)2-OH (13, 52 mg, 0.85 mmol, 1.30 equiv.), COMU (36 mg, 0.85 mmol, 1.30 equiv.) and DIPEA (15 μL, 0.85 mmol, 1.30 equiv.) in DMA (0.4 mL) was stirred for 30 min at 0 °C. The silylated monocyclic peptide was then added to the activated dihydroxyisoleucine derivative and stirred for 1 h at 0 °C and then overnight at room temperature. The mixture was then diluted with EtOAc (50 mL) and washed with 10% KHSO4 solution (3 × 5 mL). The organic layer was washed with brine (2×20 mL), dried over Na2SO4 and evaporated in vacuo. The crude product of 67 was used in the next step without further purification. HRMS(ESI):m / z calculated value C64 H 87 N7O 12 SSi2(M+H) + 1234.5745,Measurement value 1234.5745.

[0107] Monocyclic hexapeptide 68: [ka]

[0108] A solution of the fully deprotected monocyclic pentapeptide 56 (100 mg, 0.188 mmol, 1.00 equiv.) and MSA (33.2 μL, 0.207 mmol, 1.10 equiv.) in DMA (4 mL) was stirred for 2 h at 50 °C. Concurrently, a solution of Fmoc-DHIle(TBS)2-OH (13,149 mg, 0.244 mmol, 1.30 equiv.), COMU (104 mg, 0.244 mmol, 1.30 equiv.) and DIPEA (42.5 μL, 0.244 mmol, 1.30 equiv.) in DMA (1.25 mL) was stirred for 30 min at 0 °C. The silylated monocyclic peptide was then added to the activated dihydroxyisoleucine derivative and stirred for 1 h at 0 °C and then overnight at room temperature. The mixture was then diluted with EtOAc (100 mL) and washed with 10% KHSO4 solution (3 x 10 mL). The organic layer was washed with brine (2 x 25 mL), dried over Na2SO4, and evaporated under reduced pressure. The crude product 68 was used in the next step without further purification. HRMS(ESI):m / z calculated value C 57 H 81 N7O 11 SSi2(M+H) + 1128.5326,Measured value 1128.5316.

[0109] Synthesis of (9H-fluoren-9-yl)methyl (2S,4R)-4-hydroxypyrrolidine-2-carboxylate hydrochloride (2): [ka]

[0110] A solution of N-Boc-protected (2S,4R)-4-hydroxyproline 1 (5.0 g, 22 mmol, 1.0 equiv.) in DMF (20 mL) was added dropwise to a solution of 9-fluorenemethanol (8.5 mg, 43 mmol, 2.0 equiv.), EDC*HCl (8.3 g, 43 mmol, 2.0 equiv.) and DMAP (396 mg, 3.24 mmol, 0.150 equiv.) in DCM (220 mL). The reaction mixture was stirred at room temperature for 2 h. Then, 10% KHSO4 solution (50 mL) was added. The organic layer was washed with brine (50 mL) and dried over Na2SO4. Then, the solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (hexane / ethyl acetate=1:1) and treated with 4 m HCl in dioxane for 30 min. The solvent was removed under reduced pressure to give product 2 as a white solid (3.7 g, 56%). HRMS(ESI):m / z calculated value C 19 H 19 NO3 (M+H-HCl) + 310.1438,Measured value 310.1426.

[0111] Synthesis of (9H-fluoren-9-yl)methyl (2S,4R)-1-((S)-4-(allyloxy)-2-amino-4-oxobutanoyl)-4-hydroxypyrrolidine-2-carboxylate (3) [ka]

[0112] Boc-l-aspartic acid 4-allyl ester (287 mg, 1.05 mmol, 1.30 equiv), compound 2 (250 mg, 0.808 mmol, 1.0 equiv), and HATU (614 mg, 1.62 mmol, 2.0 equiv) were dissolved in DMF (2 mL) at 0 °C. Then, DIPEA (563 μL, 3.23 mmol, 4 equiv) was added and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was then diluted with EtOAc (50 mL). The organic layer was washed with 10% KHSO4 solution (2 × 10 mL), saturated NaHCO3 (10 mL), and brine (10 mL). After drying over Na2SO4 and removing the solvent under reduced pressure, the crude product was purified by silica gel column chromatography (hexane / ethyl acetate = 2:1) and then treated with 4 m HCl in dioxane for 30 min. The solvent was removed under reduced pressure to give product 3 as a yellow oil (344 mg, 85%). HRMS(ESI):m / z calculated value C 26 H 28 N2O6(M+H-HCl) + 465.2020,Measured value 465.2017.

[0113] Synthesis of monocyclic octapeptide 5: [ka]

[0114] The fully deprotected monocyclic pentapeptide 4 (50 mg, 0.09 mmol, 1.00 equiv) was dissolved in DMA (1 mL) and MSA (17 μL, 0.1 mmol, 1.1 equiv) was added. The solution was stirred for 2 h at 50 °C. A solution of Fmoc-DhI(OTBS)2-OH (75 mg, 0.12 mmol, 1.3 equiv), COMU (52 mg, 0.12 mmol, 1.3 equiv), and DIPEA (43 μL, 0.24 mmol, 2.6 equiv) in DMA (0.5 mL) was stirred for 30 min at 0 °C. The silylated monocyclic pentapeptide was added to the preactivated amino acids and stirred for 1 h. Then, H-Asp(OAll)-Hyp-OFm*HCl (70 mg, 0.14 mmol, 1.5 equiv) and HATU (53 mg, 0.14 mmol, 1.5 equiv) were added to the reaction mixture at 0°C. DIPEA (49 μL, 0.28 mmol, 3.0 equiv) was added and the reaction mixture was stirred for 2 h, diluted with EtOAc (50 mL) and washed with 0% citric acid (2 × 10 mL) and saturated NaHCO3 (2 × 10 mL). The organic layer was washed with brine (2 × 20 mL), dried over Na2SO4 and evaporated in vacuo. The crude product 5 was carried on to the next step without further purification. HRMS(ESI):m / z calculated value C 83 H 107 N9O 16 SSi2(M+H) + 1574.7167, measured value 1574.7134.

[0115] Synthesis of monocyclic C-terminally and N-terminally deprotected octapeptide 6: [ka]

[0116] The monocyclic octapeptide 5 obtained from the above reaction without further purification by chromatography was dissolved in DMF / ACN (2 mL). Et2NH (191 μL, 1.85 mmol, 20.0 equiv) was then added and stirred at room temperature for 30 min. The solvent was removed under reduced pressure and the precipitate was redissolved in THF (1 mL). A solution of TBAF in THF (1 m, 0.85 mL, 10 equiv) was then added and the reaction mixture was stirred at room temperature for 4 h. The solvent was removed in vacuo and the crude product was purified by C18 reverse phase chromatography (AcN / H2O 5%-70%) to give the product 6 as a white solid (51 mg, 64% after 4 steps). HRMS(ESI):m / z calculated value C 42 H 59 N9O 14 S (M+H) + 946.3975,Measured value 946.3972.

[0117] Synthesis of allyl ester protected deoxyamanine-precursor (7): [ka]

[0118] Monocyclic octapeptide 6 (10.0 mg, 10.6 μmol, 1.00 equiv.) was dissolved in DMF (5 mL). Then, DIPEA (3.68 μL, 21.4 μmol, 2.00 equiv.) and HATU (8.04 mg, 21.1 μmol, 2.00 equiv.) were added at 0° C. The reaction mixture was stirred for 5 h, during which the reaction mixture was allowed to warm to room temperature. The crude product was purified by preparative HPLC (Sunfire Prep C18 OBD 10 μm, 50×150 mm column, gradient A) to give the allyl-protected deoxyamanine-precursor 7 (6 mg, 68%) as a white powder. HRMS(ESI):m / z calculated value C 42 H 57 N9O 13 S (M+H) + 928.3869,Measured value 928.3885.

[0119] Synthesis of (S)-deoxyamanine (8): [ka]

[0120] Deoxyamanine precursor 7 (3.0 mg, 3.2 μmol, 1.0 equiv.) was dissolved in THF (0.3 mL). Morpholine (5.58 μL, 64.6 μmol, 20.0 equiv.) and Pd(PPh3)4 (747 μg, 0.64 μmol, 0.2 equiv.) were then added. The reaction mixture was vigorously stirred for 3 h. The crude product was purified by preparative HPLC (Sunfire Prep C18 OBD 10 μm, 50 × 150 mm column, gradient B) to give (S)-deoxyamanine (8) (1.5 mg, 50%) as a white powder. HRMS(ESI):m / z calculated value C 39 H 53 N9O 13 S (M+H) + 888.3556,Measured value 888.3529.

[0121] Prep HPLC purification gradient: Gradient A: 0-30 minutes 10%-30%B,30-35 minutes 30-100%B;40-45 minutes 100%B;45-50 minutes 100-10%B 0.1% formic acid in water (solvent A) and 0.1% formic acid in ACN (solvent B). Gradient B: 0-30 minutes 15%-40%B,30-35 minutes 40-100%B;35-40 minutes 100%B,40-15 minutes 15%B 0.1% formic acid in water (solvent A) and 0.1% formic acid in ACN (solvent B)

Claims

1. A process for preparing a compound of formula (I), comprising the steps of: 【Chemistry 1】 Where: a) a compound of formula (IIa) 【Chemistry 2】 reacting in reaction step a) with a peptide bond forming reagent; If applicable, the compound is R ＰＧＰ and / or R ＰＧＯＨ and / or R ＡＬ and / or R ＡＭＩＮ reacting with a deprotecting agent which removes During the ceremony X １ , X ２ , X ３ , and X ４ , and Y １ , Y ２ , Y ３ , and Y ４ is H, or Y １ , Y ２ , Y ３ , and Y ４ One, two, three or all of are OH or NH ２ and all other Y's are H and the corresponding X's are NHR ＡＭＩＮ OR ＰＧＰ and all other X's are H; Here, R ＰＧＰ is a protecting group for a phenolic OH group, Or, R ＡＭＩＮ is a protecting group for a phenolic amino group, or Y １ , Y ２ , Y ３ , and Y ４ and the corresponding X is F, Cl, Br, I, CN, NO ２ , Acyl, N ３ or an alkyne, and all other X and Y are H; Z and W are H, or Z is OH and W is OR ＰＧＯＨ where R ＰＧＯＨ is a protecting group for a hydroxyl group, R ＡＬ is a protected carboxyl group or a 1-5 AA peptide; V is OH or a 1-5 AA peptide; Q is S or SO; The method is carried out via step a).

2. The method of claim 1, wherein the peptide bond forming reagent is a coupling reagent selected from the group consisting of carbodiimides, imidazolinium reagents, phosphonium salts, organophosphorus reagents, uronium salts, pyridinium reagents, and phosphonic acids.

3. R PGP is a phenolic OH protecting group that is not labile to acid or alkali; The method according to claim 1 or 2.

4. R AMIN is a phenolic amino protecting group that is not acid- or alkali-labile. The method according to any one of claims 1 to 3.

5. R PGOH is a protecting group for a hydroxyl group that can be cleaved by a fluoride ion. The method according to any one of claims 1 to 4.

6. R AL is O-allyl or O-methyl ester or 1-5AA peptide; The method according to any one of claims 1 to 5.

7. V is OH. The method according to any one of claims 1 to 6.

8. The method according to any one of claims 1 to 7, Compound of formula (III) 【Chemistry 3】 and a compound of formula (IV) 【Chemistry 4】 (In the ceremony X １ , X ２ , X ３ , X ４ , W, Q, and R ＡＬ has the same meaning as defined in claim 1; R ＣＯＯＹ is a carboxyl protecting group.) wherein in reaction step (e), (III) and (IV) are reacted with a peptide bond forming reagent to obtain a compound characterized by (IIa).

9. R COOY is fluorenylmethyl or benzyl. The method according to claim 8.

10. 10. The method of claim 8 or 9, Compound of formula (V) 【Chemistry 5】 and a compound of formula (VI) 【Chemistry 6】 (In the formula, X １ , X ２ , X ３ , X ４ , Q, and W have the same meanings as defined in claim 1; R ＮＨＢ is an amino protecting group.) wherein in the reaction step (f), the amino group of (V) is preactivated, and the preactivated (V) and (VI) are reacted with a peptide bond forming reagent to obtain compound (III).

11. R NHB is an amino protecting group cleavable under alkaline conditions. The method of claim 10.

12. The peptide bond forming reagent is HATU, COMU, HBTU, TBTU, TOMBU, COMBU, or HCTU. The method according to claim 8 or 10.

13. 2. The method of claim 1 , Regarding the compound of formula (Idesox) in which Q is S obtained by reaction step a), 【Chemistry 7】 (In the formula, Y １ , Y ２ , Y ３ , Y ４ , Z, and V have the same meanings as defined in claim 1. i. using manganese ions; ii. using PPO, dibenzoyl peroxide, tert-butyl peroxybenzoate, or lauroyl peroxide; or iii. using iodine and oxygen; In the reaction step (h), the sulfur atom is oxidized to obtain a compound (Iox) of formula (I) in which Q is SO. 【Chemistry 8】

14. The use of manganese ions, 14. The method of claim 13, by reacting said compound with a compound of formula (XV) and Mn(OTf)2 and H2O2. 【Chemistry 9】

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