CN104640842B

CN104640842B - Aplysiatoxin-10 derivant, its preparation method and comprise its anticancer pharmaceutical composition

Info

Publication number: CN104640842B
Application number: CN201380048896.4A
Authority: CN
Inventors: 朴永浚; 郑镇姣; 崔英美; 李玟燮; 崔俊宪; 赵银珠; 宋贤南; 朴星俊; 李锺侠; 洪承绪
Original assignee: Match Imec Inter Uni Micro Electr
Current assignee: Match Imec Inter Uni Micro Electr
Priority date: 2012-09-20
Filing date: 2013-09-16
Publication date: 2016-11-30
Anticipated expiration: 2033-09-16

Abstract

The invention provides aplysiatoxin 10 derivant with excellent antitumor activity, produce the method for aplysiatoxin 10 derivant and comprise aplysiatoxin 10 derivant anticancer pharmaceutical composition as active component.

Description

Dolastatin-10 derivatives, preparation method thereof and anticancer pharmaceutical composition comprising the same

Technical Field

The present invention relates to dolastatin-10 derivatives having excellent anticancer activity, a method for producing dolastatin-10 derivatives, and an anticancer pharmaceutical composition comprising dolastatin-10 derivatives as an active ingredient.

Background

Dolastatin-10 was isolated from the Indian ocean mollusk sea hare (Dolabella auricularia) in 1987 and was identified as having the following chemical structure [ J.Am.chem.Soc.,1987,109, 6883-. Dolastatin-10 shows similar physiological activity as paclitaxel and vinca alkaloids as antimicrotubule agents, but is a structurally different peptide [ chem.ind.,1999, 51-55; curr. pharm. Des.,1999,5,139-162 ].

Dolastatin 10 is expected to have no side effects known for paclitaxel and vinca alkaloids, such as bone marrow toxicity, due to differences in structure from paclitaxel and vinca alkaloids and demonstrated inhibitory activity in vitro in model experiments in various tumors and clinical trials in humans. However, despite the different structures, dolastatin-10, similar to paclitaxel and vinca alkaloids, has been found in clinical studies to have myelotoxicity, neurotoxicity and other side effects [ clin. cancer res.,2000,6, 1293-; drugs of the Future,1999,24(4),404- > 409 ].

Various dolastatin-10 derivatives have been synthesized, which are derivatives of Dov (dolavine), Val (valine), dolalieucine (dolasioleucine), Dolaproline (Dolaproline), and dolamin (dolamine) in their 5 parts. While dolastatin-10 derivatives are disclosed in WO 2003/008378, there has been concentrated research on derivatives of dolastaline and doramexane moieties in which the methoxy group of the dolastaline moiety is replaced by a thiomethoxy group, and in u.s patent 5599902, the doramexane moiety has been modified. Meanwhile, EP260858 discloses a derivative in which Dov part of dimethylvaline is replaced by monomethylvaline.

Disclosure of Invention

Technical problem

The present invention has been made by extensive and intensive studies on dolastatin-10 derivatives having more potent cytotoxicity to cancer cells, and it has been found that dolastatin-10 derivatives represented by the following chemical formula I, in which the pyrrolidine ring of dolastatin moiety is modified, exhibit excellent anticancer activity.

Accordingly, it is an object of the present invention to provide a dolastatin 10 derivative of formula I or a pharmaceutically acceptable salt thereof, which has excellent anticancer activity.

It is another object of the present invention to provide a method for producing dolastatin 10 derivatives of formula I or pharmaceutically acceptable salts thereof.

Still another object of the present invention is to provide an anticancer pharmaceutical composition comprising dolastatin 10 derivatives of formula I or pharmaceutically acceptable salts thereof.

Technical scheme

According to one aspect of the present invention, there is provided dolastatin-10 derivatives of formula I:

[ chemical formula I ]

Wherein,

R₁、R₂、R₃、R₄and R₅Each independently is hydrogen or C₁-C₄An alkyl group;

R₆is hydrogen, hydroxy, C₁-C₄Alkoxy, amino, oxo (═ O), or oximino (═ N-OH);

ar is aryl;

x is a carbon atom, an oxygen atom or a sulfur atom; and

when X is a carbon atom, R₇Is hydroxy, amino, C₁-C₄Alkoxy radical, C₁-C₄Alkylamino or oxo (═ O); when X is an oxygen atom or a sulfur atom, R is absent₇。

As used herein, the term "C₁-C₄Alkyl "is intended to encompass straight or branched chain hydrocarbon groups of 1 to 4 carbon atoms, including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

As used herein, the term "aryl" is intended to encompass all aryl, heteroaryl, and partially reduced derivatives thereof. The term "aryl" refers to a 5 to 15 membered monocyclic or fused ring, and the term "heteroaryl" refers to an aromatic group having at least one heteroatom, such as oxygen, sulfur or nitrogen. Representative examples of aryl groups include, but are not limited to: phenyl, naphthyl, pyridyl, furyl, thienyl, indolyl, quinolinyl, imidazolinyl, oxazolyl, thiazolyl, and tetrahydronaphthyl.

As used herein, the term "C₁-C₄Alkoxy "is intended to encompass straight or branched chain alkoxy groups of 1 to 4 carbon atoms, and includes, but is not limited to: methoxy, ethoxy and n-propoxy.

As used herein, the term "C₁-C₄Alkylamino means having C₁-C₄Amino for alkyl substituents, and include, but are not limited to: methylamino, ethylamino, and n-propylamino.

At C₁-C₄Alkyl, aryl, C₁-C₄Alkoxy and C₁-C₄In the alkylamino radical, at least one hydrogen may be replaced by C₁-C₄Alkyl radical, C₂-C₄Alkenyl radical, C₂-C₄Alkynyl, C₃-C₁₀Cycloalkyl radical, C₁-C₄Haloalkyl, C₁-C₄Alkoxy radical, C₁-C₄Thioalkoxy, aryl, acyl, hydroxy, thioSubstituted by halogen, amino, alkoxycarbonyl, carboxyl, carbamoyl, cyano or nitro.

In a preferred embodiment of the dolastatin 10 derivatives according to the invention,

R₁、R₃and R₄Each is C₁-C₄An alkyl group;

R₂and R₅Each independently is hydrogen or C₁-C₄An alkyl group;

ar is unsubstituted phenyl or is selected from the group consisting of C₁-C₄Alkyl radical, C₁-C₄Phenyl substituted with at least one of the group consisting of alkoxy and halogen;

x is a carbon atom; and

R₇is hydroxy, amino, C₁-C₄Alkoxy or C₁-C₄An alkylamino group.

In a more preferred embodiment of the dolastatin 10 derivative according to the invention,

R₁、R₃and R₄Each is methyl;

R₂and R₅Each independently hydrogen or methyl;

R₆is hydrogen, hydroxy, methoxy, amino, oxo (═ O) or oximino (═ N-OH);

ar is unsubstituted phenyl or phenyl substituted with at least one selected from the group consisting of methyl, methoxy and halogen;

x is a carbon atom; and

R₇is hydroxyl, amino, methoxy or N-methylamino.

All stereoisomers of dolastatin-10 derivatives, including compounds of the following formula Ia, are within the scope of the present invention.

[ chemical formula Ia ]

Wherein,

R₁、R₂、R₃、R₄、R₅、R₆ar, X and R₇Respectively, as defined in formula I.

In the formula Ia, R₅、R₆And R₇Each independently in the (R) -configuration, (S) -configuration or racemic mixture.

As used herein, the term "pharmaceutically acceptable salt" is intended to encompass organic or inorganic non-toxic salts and includes, but is not limited to, for example, hydrochloride, sulfate, nitrate, phosphate, acetate, benzene sulfate, citrate, and the like.

Specific examples of dolastatin-10 derivatives according to the present invention include:

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino (butanamido)) butyramide (I-1);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((3-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-2);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((4-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-3);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2, 4-dichloro-5-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-4);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- ((2- (4-methoxyphenyl) -2-oxoethyl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-5);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (((R) -1-oxo-1-phenylpropan-2-yl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-6);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (((S) -1-oxo-1-phenylpropan-2-yl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-7);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -4-hydroxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-8);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -3-methoxy-1- ((R) -2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) -4-oxopyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-9);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4S) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-10);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-11);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-12);

(S) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (2-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-13);

(S) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (3-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-14);

(S) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (4-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-15);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((S) -1- (3, 5-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-16);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((S) -1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-17);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-18);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R, Z) -1- (hydroxyimino) -1- (3-methoxyphenyl) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-19);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (hydroxyimino) -1- (3-methoxyphenyl) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-20);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (4-fluorophenyl) -1- (hydroxyimino) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-21);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (5-bromo-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-22);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutyramide (I-23);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-24);

(S) -N- ((3R,4S,5S) -1- ((2S,4R) -2- ((1R,2R) -3- (((E) -2- (2-fluoro-4-methoxyphenyl) -2-hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-25);

(S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-26);

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-27);

(S) -N- ((3R,4S,5S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2- (o-tolyl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-28);

(S) -N- ((3R,4S,5S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2- (p-tolyl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-29);

(S) -N- ((3R,4S,5S) -1- ((2S,4R) -2- ((1R,2R) -3- (2- (4-fluorophenyl) -2-oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-30);

(S) -N- ((3R,4S,5S) -1- ((2S,4R) -2- ((1R,2R) -3- (2- (2-fluoro-4-methoxyphenyl) -2-oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-31);

(S) -N- ((3R,4S,5S) -1- ((S) -4- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) thiazolidin-3-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyramido) butanamide (I-32);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4S) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-33);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2-phenylethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-34);

(2S) -N- ((3R,4S) -1- ((2S,4R) -2- ((1R,2R) -3- (2- (2-hydroxy-2-phenylethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-35);

(2S) -N- ((3R,4S) -1- ((2S) -2- ((1R,2R) -3- (((2R) -1-amino-1-phenylpropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-36);

(2S) -N- ((3R,4S) -1- ((2S,4R) -2- ((1R,2R) -3- (((2S) -1-amino-1-phenylpropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-37);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2- (pyridin-2-yl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-38);

(2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2- (thien-2-yl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-39);

(2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-40);

(2S) -2- ((S) -2- (dimethylamino) -3-methylbutanamino) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-41);

(2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-42);

(2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-43);

(2S) -N- ((3R,4S) -1- ((2S,4S) -4-amino-2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) pyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-44); and

(2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4- (methylamino) pyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-45).

According to another aspect of the present invention, there is provided a method for producing dolastatin-10 derivatives represented by the following chemical formula I, which comprises: subjecting a compound represented by the following chemical formula II and a compound represented by the following chemical formula III to a condensation reaction, and deprotecting the condensation product when a protecting group is present.

[ chemical formula II ]

[ chemical formula III ]

Wherein,

R₁and R₂Each independently is hydrogen, C₁-C₄An alkyl or amino protecting group;

R₃、R₄and R₅Each independently is hydrogen or C₁-C₄An alkyl group;

R₆is hydrogen, protected or unprotected hydroxy, C₁-C₄Alkoxy, protected or unprotected amino, oxo (═ O), or oximo (═ N-OH);

ar is aryl;

x is a carbon atom, an oxygen atom or a sulfur atom; and

when X is a carbon atom, R₇Being a protected or unprotected hydroxyl group, a protected or unprotected amino group, C₁-C₄Alkoxy, protected or unprotected C₁-C₄Alkylamino or oxo (═ O); when X is an oxygen atom or a sulfur atom, R is absent₇。

As the amino-protecting group, a t-butyloxycarbonyl group (t-Boc), a benzyloxycarbonyl group (Cbz), a fluorenylmethyloxycarbonyl group (Fmoc) or a benzyl group (Bn) can be used, but not limited thereto.

The hydroxyl group can be protected using a t-butyldimethylsilyl group, but is not limited thereto.

The compound of formula III may be in the form of a salt, including but not limited to hydrochloride and Trifluoroacetate (TFA).

The condensation reaction may be carried out in the presence of a condensing agent. Condensing agents that may be used include, but are not limited to, Dicyclohexylcarbodiimide (DCC), diphenylphosphoryl azide (DPPA), diethyl cyanophosphate (DEPC), and benzotriazol-1-yl-oxy-tris (dimethylamino) phosphonium hexafluorophosphate (BOP reagent).

For the condensation reaction, if necessary, an organic base such as triethylamine and Diisopropylethylamine (DIPEA) may also be used together with a condensing agent.

As the reaction solvent, halogenated aliphatic hydrocarbons such as chloroform and dichloromethane; ethyl acetate; tetrahydrofuran (THF); dimethylformamide (DMF) and/or acetonitrile, and preferably, the reaction temperature may be set to 0 to 50 ℃, and, more preferably, 0 ℃ to room temperature.

Deprotection of the amino protecting group can be carried out using 10% palladium on carbon or 20% palladium hydroxide.

Deprotection of the hydroxyl protecting group can be performed using tetrabutylammonium fluoride.

As previously described in [ u.s. patent 5,654,399; tetrahedron Letters, Vol.32, No.21, pp 2395-2398] to synthesize the compounds of formula II.

Meanwhile, the compound of chemical formula III is synthesized according to the following reaction scheme I, which is a modification of the previously disclosed method [ Tetrahedron, Vol.49, No.9, pp.1913-1924 ]. The processes described in the following reaction flow charts are exemplary illustrations, and the process order, reaction reagents, conditions, and the like may be modified as necessary.

[ reaction scheme I ]

Wherein,

R₄and R₅Each independently is hydrogen or C₁-C₄An alkyl group;

ar is aryl;

x is a carbon atom, an oxygen atom or a sulfur atom;

when X is a carbon atom, R₇Being a protected or unprotected hydroxyl group, a protected or unprotected amino group, C₁-C₄Alkoxy, protected or unprotected C₁-C₄Alkylamino or oxo (═ O); when X is an oxygen atom or a sulfur atom, R is absent₇(ii) a And

R₈is C₁-C₄An alkyl group.

As shown in reaction scheme I, the compound of formula VI is synthesized by reacting the compound of formula IV with the compound of formula V in the presence of triethylamine as a base and dibutylboron trifluoromethanesulfonate as a lewis acid. In this aspect, the stereochemistry of the compound of formula VI can be controlled depending on the order of addition of the base and lewis acid.

Subsequently, the compound of formula VI is methylated using trimethyloxonium tetrafluoroborate in the presence of 1, 8-bis (dimethylamino) naphthalene (proton sponge) to give the compound of formula VII, which is subsequently converted to the compound of formula VIII in the presence of hydrogen peroxide and lithium hydroxide.

Subsequently, the compound of formula VIII is coupled with the compound of formula IX to give the compound of formula X. Salt forms of the compounds of formula IX may also be used, including, but not limited to, hydrochloride, Trifluoroacetate (TFA), and the like.

For the condensation reaction (coupling), the same reagents and conditions as those for the condensation reaction between the compound of formula II and the compound of formula III may be used.

Finally, deprotection of the amino protecting group of the compound of formula X with hydrochloric acid or trifluoroacetic acid yields the compound of formula III.

The dolastatin-10 derivatives of the present invention exhibited excellent antitumor activity (test example 1).

Thus, it is contemplated according to another aspect of the present invention to be an anticancer agent comprising a dolastatin-10 derivative of formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, which is particularly useful for the treatment of breast cancer.

The anticancer agent according to the present invention may be administered parenterally (e.g., orally or by inhalation) or parenterally (e.g., by injection, inhalation, deposition (deposton), transplantation, suppository). For injection, administration can be intravenous, subcutaneous, intramuscular, or intraperitoneal. The anticancer agent of the present invention may be formulated into tablets, capsules, granules, fine granules, powders, sublingual tablets, suppositories, pastes, injections, emulsions, suspensions, syrups, sprays, etc. according to the administration route. When the anticancer agent of the present invention is formulated into various preparations, conventional pharmaceutically acceptable carriers for each preparation may be used. Examples of pharmaceutically acceptable carriers include: excipients, binders, disintegrants, lubricants, preservatives, antioxidants, isotonicity agents, buffers, coating agents, sweeteners, solubilizers, bases, dispersants, wetting agents, suspending agents, stabilizers, colorants, and the like.

In the anticancer drug according to the present invention, although the content of the compound of the present invention or a pharmaceutically acceptable salt thereof varies depending on the formulation, it ranges from about 0.01 wt% to 95 wt%.

The effective dose of the anticancer agent of the present invention depends on various factors including the kind, body weight, sex and severity of diseases of a mammalian (including human) subject. In general, the compounds according to the invention can be administered via the parenteral route in daily doses ranging from 0.01 to 50mg/kg of body weight and via the parenteral route in daily doses ranging from 0.01 to 10mg/kg of body weight. The compounds may be administered in single or multiple doses per day, as prescribed by a physician or pharmacist.

Advantageous effects

Dolastatin-10 derivatives according to the present invention are novel compounds that can be effectively used for the treatment of cancer and the inhibition of tumor growth. For example, the compounds effectively inhibit or prevent the growth of precancerous and malignant cells, and have been found to be effective for the treatment of blood cancers as well as cancer solid tumors, particularly colorectal cancer, lung cancer, breast cancer, gastric cancer, cervical cancer and bladder cancer.

Detailed Description

The invention will be better understood from the following examples which are given by way of illustration and are not intended to limit the invention.

Preparation example 1: preparation of the Compound of formula IV

Preparation examples 1 to 1: N-Boc-L-prolinaldehyde (IV-1)

A solution of N-tert-butoxycarbonyl-L-prolinol (2g, 9.9mmol) in 12mL of dimethyl sulfoxide was cooled to 5-10 ℃ and mixed with triethylamine (4.8mL, 34.7mmol) at the same temperature with stirring for 15 minutes. Sulfur trioxide-pyridine complex (5.5g, 34.7mmol) was added to the reaction solution at 0 ℃ followed by stirring for 2 hours.

After the reaction was complete, 50mL of water was added to the reaction mixture, which was then extracted 3 times with 20mL of dichloromethane. The resulting organic layers were combined and washed with 50mL of 50% aqueous citric acid solution, 50mL of water and 50mL of saturated sodium bicarbonate, followed by drying over anhydrous magnesium sulfate and concentration under vacuum. The residue was isolated and purified by column chromatography to give the title compound: 1.87g (94%).

1H NMR(400MHz,CDCl3):1.43～1.46(m,9H),2.18～1.79(m,4H),3.59～3.40(m,2H),4.05～4.20(m,1H),9.46～9.56(s,1H)。

Preparation examples 1 to 2: N-Boc-trans-4-methoxy-L-prolinaldehyde (IV-2)

The same procedure as in preparation example 1-1 was repeated except that N-t-butoxycarbonyl-L-prolinol was not used but N-t-butoxycarbonyl-trans-4-methoxy-L-prolinol was used to obtain the title compound: 40.8g (97%).

1H NMR(400MHz,CDCl3):1.45(s,9H),1.96～1.98(m,1H),2.1～2.2(m,1H),3.29(S,3H),3.51～3.53(m,1H),3.71～3.73(m,1H),3.93～3.94(m,1H),4.19～4.29(m,1H),9.43(S,1H)。

Preparation examples 1 to 3: N-Boc-cis-4-methoxy-L-prolinaldehyde (IV-3)

The same procedure as in preparation example 1-1 was repeated except that N-t-butoxycarbonyl-L-prolinol was used instead of N-t-butoxycarbonyl-cis-4-methoxy-L-prolinol to obtain the title compound: 6.94g (65%).

1H NMR(400MHz,CDCl3):1.43(s,9H),2.04～2.17(m,1H),2.7(m,1H),3.24(S,3H),3.43～3.47(m,1H),3.56～3.71(m,1H),3.89～3.91(m,1H),4.05～4.18(m,1H),9.51(S,1H)。

Preparation examples 1 to 4: N-Boc-cis-4- (tert-butyldimethylsilyl) oxy-L-prolinaldehyde (IV-4)

The same procedure as in preparation example 1-1 was repeated except that N-t-butoxycarbonyl-cis-4- (t-butyldimethylsilyl) oxy-L-prolinol was used instead of N-t-butoxycarbonyl-L-prolinol to obtain the title compound: 3.8g (52%).

1H NMR(400MHz,CDCl3):0.04(s,3H),0.07(s,3H),0.85(s,9H),1.45(s,9H),2.04～2.08(m,1H),2.15～2.24(m,1H),3.38～3.52(m,2H),4.06～4.20(m,1H),4.36～4.36(m,1H),9.55(s,1H)。

Preparation examples 1 to 5: N-Boc-trans-4- (tert-butyldimethylsilyl) oxy-L-prolinaldehyde (IV-5)

The same procedure as in preparation example 1-1 was repeated except that N-t-butoxycarbonyl-L-prolinol was used instead of N-t-butoxycarbonyl-trans-4- (t-butyldimethylsilyl) oxy-L-prolinol to obtain the title compound: 9.64g (66%).

1H NMR(400MHz,CDCl3):0.07(s,6H),0.87(s,9H),1.45(s,9H),1.89～2.08(m,2H),3.34～3.56(m,2H),4.12～4.37(m,2H),9.43(S,1H)。

Preparation examples 1 to 6: (S) -tert-butyl-4-formylthiazolidine-3-carboxylate (IV-6)

The same procedure as in preparation example 1-1 was repeated except that (R) -tert-butyl-4- (hydroxymethyl) thiazolidine-3-carboxylate was used instead of N-tert-butoxycarbonyl-L-prolinol to obtain the title compound: 9.21g (44%).

1H NMR(400MHz,CDCl3):1.47(s,9H),3.16～3.23(m,2H),4.27～4.65(m,3H),9.57(s,1H)。

Preparation example 2: preparation of the Compound of formula VI

Preparation example 2-1: (S) -tert-butyl 2- ((1R,2R) -1-hydroxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylic acid ester (VI-1)

A solution of (R) -4-phenyl-3-propionyloxyoxazolidin-2-one (605mg, 2.7mmol) in 8mL of dichloromethane was cooled to 0 deg.C, followed by the addition of triethylamine (0.5mL,3.7 mmol). Then, dibutylboron trifluoromethanesulfonate was added dropwise, and after dropwise adding dibutylboron trifluoromethanesulfonate with stirring, stirring was performed for 45 minutes. The resulting solution was cooled to-78 ℃, followed by dropwise addition of a solution of the compound (IV-1) (500mg, 2.5mmol) obtained in production example 1-1 in 5mL of dichloromethane, and stirring for 1 hour. Stirring is carried out at 0 ℃ for 1 hour, and at 20-25 ℃ for 1 hour.

The reaction was stopped with phosphate buffer (pH 7.2, 3mL) and 9mL of methanol. After cooling the reaction mixture to 0 ℃, methanol: a mixture of 33% hydrogen peroxide (2:1, 9mL) was added dropwise to the mixture. Followed by stirring for 1 hour and concentration under vacuum. The concentrate was then dissolved in ether and washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under vacuum to give a yellow oil. Recrystallization from ethyl acetate gave the title compound as a white solid: 0.59g (59%).

MS(EI)m/z:419[M+H]+。

1H NMR(400MHz,CDCl3):1.10(m,3H),1.39(s,9H),1.67～1.84(m,7H),3.01(m,1H),3.67～3.81(m,3H),4.14(m,1H),4.70(m,1H),5.03(m,1H),5.43(m,1H),7.26～7.41(m,5H)。

Preparation examples 2 to 2: (2S,4R) -tert-butyl-2- ((1R,2R) -1-hydroxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) -4-methoxypyrrolidine-1-carboxylic acid ester (VI-2)

The same procedures as in preparation example 2-1 were repeated except that the compound (IV-1) obtained in preparation example 1-1 was not used but the compound (IV-2) obtained in preparation example 1-2 was used to obtain 3.6g (28%) of the title compound.

1H NMR(400MHz,CDCl3):1.24(m,3H),1.47(m,9H),1.98(m,1H),2.25(m,1H),3.28(s,3H),3.35(m,1H),3.48(m,3H),3.61(m,1H),3.92～4.07(m,4H),4.24(m,1H),4.69(m,1H),5.42(m,1H),7.37(m,5H)。

Preparation examples 2 to 3: (2S,4R) -tert-butyl-4- ((tert-butyldimethylsilyl) oxy) -2- ((1R,2R) -1-hydroxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylic acid ester (VI-3)

The same procedure as in preparation example 2-1 was repeated, except that the compound (IV-1) obtained in preparation example 1-1 was not used, but the compound (IV-5) obtained in preparation example 1-5 was used, to obtain the title compound: 8.9g (71%).

1H NMR(400MHz,CDCl3):0.05(s,6H),0.86(s,9H),1.22～1.25(m,6H),1.50(s,9H),1.87(m,1H),2.16～2.18(m,1H),3.22～3.25(m,1H),3.43～3.45(m,1H),3.61(m,1H),3.91～3.92(m,1H),4.13(m,1H),4.25～4.27(m,1H),4.33(m,1H),4.66～4.70(m,1H),5.41～5.45(m,1H),7.26～7.39(m,5H)。

Preparation example 3: preparation of the Compound of formula VII

Preparation example 3-1: (S) -tert-butyl-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylic acid ester (VII-1)

The compound (VI-1) (5g, 12.0mmol) obtained in production example 2-1 andthe molecular sieve was stirred and mixed in 100mL of dichloromethane for 15 minutes. The resulting solution was cooled to 0 deg.C, then proton sponge (6.7g,31.0mmol) and trimethyloxonium tetrafluoroborate (4.4g,30.0mmol) were added and stirred for 2 hours. After raising the temperature to 20-25 ℃, stirring was continued for an additional 46 hours.

When the reaction was complete, filtration through celite was followed by vacuum concentration. The residue was separated and purified by column chromatography (n-hexane: ethyl acetate ═ 5:1) to give the title compound: 5.1g (57%).

MS(EI)m/z:433[M+H]+。

1H NMR(400MHz,CDCl3):1.21(m,3H),1.45(m,9H),1.79～1.91(m,3H),3.20-3.55(m,5H),3.80(m,2H),4.09(m,2H),4.24(m,1H),4.66(m,1H),5.43(m,1H),7.36(m,5H)。

Preparation examples 3 to 2: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylic acid ester (VII-2)

The same procedure as in preparation example 3-1 was repeated, except that the compound (VI-1) obtained in preparation example 2-1 was not used, but the compound (VI-2) obtained in preparation example 2-2 was used, to obtain the title compound: 1.6g (45%).

1H NMR(400MHz,CDCl3):1.21(m,3H),1.47(m,9H),1.92(m,1H),2.16(m,1H),3.33(m,3H),3.47(m,3H),3.76-4.10(m,6H),4.24(m,1H),4.66(m,1H),5.30-5.39(m,1H),7.33(m,5H)。

Preparation examples 3 to 3: (2S,4R) -tert-butyl-4- ((tert-butyldimethylsilyl) oxy) -2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylic acid ester (VII-3)

The same procedure as in preparation example 3-1 was repeated, except that the compound (VI-1) obtained in preparation example 2-1 was not used, but the compound (VI-3) obtained in preparation example 2-3 was used, to obtain the title compound: 6.5g (71%).

1H NMR(400MHz,CDCl3):0.05(s,6H),0.86(s,9H),1.18～1.20(d,3H),1.45～1.49(d,9H),1.87(m,1H),2.16～2.18(m,1H),3.29～3.30(m,1H),3.41(s,3H),3.69～3.80(m,1H),3.89～3.93(m,1H),3.99～4.07(m,1H),4.23～4.26(m,1H),4.38(m,1H),4.63～4.66(m,1H),5.38～5.49(m,1H),7.26～7.39(m,5H)。

Preparation example 4: preparation of the Compound of formula VIII

Preparation example 4-1: (2R,3R) -3- ((S) -1- (tert-Butoxycarbonyl) pyrrolidin-2-yl) -3-methoxy-2-methylpropanoic acid (VIII-1)

A solution of the compound (VII-1) (5.7g, 13.2mmol) obtained in production example 3-1 in 65mL of tetrahydrofuran was cooled to 0 ℃, 30% hydrogen peroxide (6.6mL, 66.0mmol) and 0.4N lithium hydroxide (66mL,26.4mmol) were added, and stirred for 3 hours.

The reaction was completed with 1M sodium sulfite (72.6mL,72.6mmol) and the reaction mixture was stirred at 20-25 ℃ for 16 hours followed by extraction with saturated sodium bicarbonate and dichloromethane cooled to 0-5 ℃. The aqueous layer was adjusted to pH 2 using 1N HCl, followed by extraction three times with ethyl acetate. The resulting organic layers were combined, dried over anhydrous magnesium sulfate, and concentrated under vacuum to give the title compound: 3.8g (99%).

[α]D 25＝-57°(c＝1,MeOH)。

1H NMR(400MHz,CDCl3):1.28(m,3H),1.45(m,9H),1.75(m,1H),1.95(m,1H),2.50(m,1H),3.25(m,1H),3.45(s,3H),3.97(m,1H),3.80-3.98(m,2H),11.1(br,1H)。

Preparation example 4-2: (2R,3R) -3- ((2S,4R) -1- (tert-Butoxycarbonyl) -4-methoxypyrrolidin-2-yl) -3-methoxy-2-methylpropanoic acid (VIII-2)

The same procedure as in preparation example 4-1 was repeated, except that the compound (VII-1) obtained in preparation example 3-1 was not used, but the compound (VII-2) obtained in preparation example 3-2 was used, to obtain the title compound: 1.5g (93%).

1H NMR(400MHz,CDCl3):1.28(m,3H),1.47(m,9H),1.99-2.55(m,4H),3.30(s,3H),3.46(s,3H),3.55～4.15(m,4H)。

Preparation examples 4 to 3: (2R,3R) -3- ((2R,4R) -1- (tert-Butoxycarbonyl) -4- ((tert-butyldimethylsilyl) oxy) pyrrolidin-2-yl) -3-methoxy-2-methylpropanoic acid (VIII-3)

The same procedure as in preparation example 4-1 was repeated, except that the compound (VII-1) obtained in preparation example 3-1 was not used, but the compound (VII-3) obtained in preparation example 3-3 was used, to obtain the title compound: 1.04g (100%).

1H NMR(400MHz,CDCl3):0.05(s,6H),0.86(s,9H),1.26(d,3H),1.48(s,9H),1.84(m,1H),2.07(m,1H),2.45～2.57(m,1H),3.27～3.30(m,1H),3.34～3.53(m,1H),3.46(S,3H),3.86～3.87(m,1H),4.05(m,1H),4.37(m,1H)。

Preparation examples 4 to 4: (2R,3R) -3- ((2S,4S) -1- (tert-Butoxycarbonyl) -4-hydroxypyrrolidin-2-yl) -3-methoxy-2-methylpropanoic acid (VIII-4)

Pyridine hydrofluoric acid (1.0mL, 69.3mmol) was slowly added to a solution of the compound (VII-3) (6.4g, 11.55mmol) obtained in production example 3-3 in 50mL of tetrahydrofuran, while stirring at 20 to 25 ℃ for 3 hours. After the reaction was complete, 50mL of saturated aqueous sodium bicarbonate solution was added to the reaction mixture, and then extracted with 50mL of ethyl acetate. The organic layer thus formed was subsequently washed with 50mL of 1N HCl and dried over anhydrous sodium sulfate and under vacuum to give (2S,4R) -tert-butyl-4-hydroxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylate: 5.4g (104%).

The concentrate was dissolved in 120mL tetrahydrofuran under an argon atmosphere, followed by addition of triphenylphosphine (4.71g, 17.75mmol) and cooling to 0-5 ℃. The reaction solution was mixed with formic acid (0.67ml,17.75mmol), followed by slowly dropwise addition of isopropyl azodicarboxylate (3.5g,17.75 mmol). Next, the mixture was stirred at 0 to 5 ℃ for 30 minutes and then at 20 to 25 ℃ for 16 hours. When the reaction was complete, the reaction mixture was concentrated under vacuum to give (2S,4S) -tert-butyl-4- (formyloxy) -2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) pyrrolidine-1-carboxylate.

The residue was cooled to 0-5 deg.C, then hydrogen peroxide (11.1ml, 115.5mmol) was slowly added dropwise, followed by 1M lithium hydroxide (2.42g, 57.8mmol) was slowly added dropwise. The reaction solution was stirred at 0-5 ℃ for 4 hours, followed by removal of tetrahydrofuran by vacuum concentration. The residue was extracted with 100mL of dichloromethane and 100mL of water. The aqueous layer was separated and adjusted to pH 2 with 1N HCl, followed by extraction twice with 100mL of ethyl acetate. The resulting several layers were combined and dried over magnesium sulfate and concentrated under vacuum to give the title compound: 3.18g (91%).

1H NMR(400MHz,CDCl3):1.34(d,3H,J＝7.2Hz),1.47(m,9H),1.97～2.00(m,1H),2.19～2.23(m,1H),2.54～2.57(m,1H),3.40～3.59(m,2H),3.49(s,3H),3.87～3.99(m,1H),4.08～4.15(m,1H),4.24～4.28(m,1H)。

LC-MS m/z:302.1[M-H]。

Preparation examples 4 to 5: (2R,3R) -3- ((2S,4S) -4-azido-1- (tert-butoxycarbonyl) pyrrolidin-2-yl) -3-methoxy-2-methylpropanoic acid (VIII-5)

The same procedures as in preparation examples 4-4 were repeated except that isopropyl azodicarboxylate was not used and diphenyl azidophosphate was used to obtain the title compound: 4.94g (90%).

[α]D 25＝-13.6°(c＝1,MeOH)。

1H NMR(400MHz,CDCl3):1.28(d,3H,J＝6.8Hz),1.47(s,9H),2.05(m,1H),2.23～2.30(m,1H),2.46～2.48(m,1H),3.05(m,1H),3.48(s,3H),3.88～3.04(m,3H)。

Preparation examples 4 to 6: (2R,3R) -3- ((S) -3- (tert-Butoxycarbonyl) thiazolidin-4-yl) -3-methoxy-2-methylpropanoic acid (VIII-6)

The same procedure as in production example 4-1 was repeated except that the compound (VII-1) obtained in production example 3-1 was not used but (R) -tert-butyl-4- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((R) -2-oxo-4-phenyloxazolidin-3-yl) propyl) thiazolidine-3-carboxylate was used to obtain the title compound: 0.89g (100%).

1H NMR(400MHz,CDCl3):1.17～1.25(d,3H),1.47(s,9H),2.65～3.12(m,4H),3.42(S,3H),3.77～4.18(m,2H),4.16～4.21(m,2H)。

Preparation example 5: preparation of the Compound of formula IX

Preparation example 5-1: 2- (2, 6-difluorophenyl) ethylamine (IX-1)

2- (2, 6-difluorophenyl) acetonitrile (2.34g,15.28mmol) was added to a solution of potassium borohydride (3.29g,61.12mmol) and Raney nickel (0.897g,15.28mmol) in 50mL of pure ethanol with stirring. Subsequently, vigorous stirring was continued at room temperature for 5 hours.

After the reaction was complete, the reaction mixture was filtered through celite and concentrated under vacuum. The residue was then extracted with ether and water and washed with brine. The organic layer was dried over anhydrous magnesium sulfate to give the title compound as a dark yellow oil.

1H NMR(400MHz,CDCl3):2.88(m,2H),2.98(m,2H),5.11(br,1H),7.32(m,2H),7.59(m,1H)。

Preparation examples 5 to 2: 2- (3-fluorophenyl) ethylamine (IX-2)

The same procedure as in preparation example 5-1 was repeated except that 2- (2, 6-difluorophenyl) acetonitrile was not used but 2- (3-fluorophenyl) acetonitrile was used to obtain the title compound.

1H NMR(400MHz,CDCl3):2.88(m,2H),2.98(m,2H),5.11(br,1H),6.83(m,1H),7.06(m,2H),7.38(m,1H)。

Preparation examples 5 to 3: 2- (2, 4-dichloro-5-fluorophenyl) ethylamine (IX-3)

The same procedure as in preparation example 5-1 was repeated except that 2- (2, 6-difluorophenyl) acetonitrile was not used but 2- (2, 4-dichloro-5-fluorophenyl) acetonitrile was used to obtain the title compound.

1H NMR(400MHz,CDCl3):2.83(m,2H),2.98(m,2H),5.11(br,1H),6.71(m,1H),7.62(m,1H)。

Preparation examples 5 to 4: 2-amino-1- (2-fluoro-4-methoxyphenyl) ethanone (IX-4)

Triphenylphosphine (6.65g,25.36mmol) and p-toluenesulfonic acid (14.5g,76.11mmol) were added to a solution of 2-azido-1- (2-fluoro-4-methoxyphenyl) ethanone (4.80g,25.36mmol) in 100mL of tetrahydrofuran, followed by stirring at room temperature for 16 h.

At the end of the reaction a precipitate formed, which was filtered and washed with cold tetrahydrofuran, followed by drying at 35 ℃ to give the title compound as a white solid: 5.12g (61%).

1H NMR(400MHz,DMSO-d6):3.83(s,3H),4.33(s,2H),6.92(m,1H),7.06(m,1H),7.88(m,1H),7.91(m,2H),8.17(br,1H)。

Preparation examples 5 to 5: (R) -tert-butyl [1- (4-methoxyphenyl) -1-oxopropan-2-yl ] carbamate (IX-5)

A solution of (R) -tert-butyl (1- (methoxy (methyl) amino) -1-oxopropan-2-yl) carbamate (5.0g,21.5mmol) in 50mL tetrahydrofuran was cooled to-20 ℃. 4-Methoxyphenylmagnesium bromide was prepared by stirring magnesium (2.1g,86.1mmol), 4-methoxybromobenzene (16.1g,86.1mmol) and a catalytically effective amount of iodine in 100mL of tetrahydrofuran, which was then added to the above solution. The reaction is carried out for 16 hours at 20-25 ℃ while stirring.

When the reaction was complete, 100mL of 1N HCl was added to give two separate layers. The aqueous layer was extracted three times with 100mL of ethyl acetate. The organic layers were combined, dried over anhydrous magnesium sulfate, and concentrated under vacuum. The residue was isolated and purified by column chromatography to give the title compound: 6.2g (66.2%).

1H NMR(400MHz,CDCl3):1.40(d,3H),1.46(s,9H),3.88(s,3H),5.24(m,1H),5.62(br,1H),6.96(d,2H),7.96(d,2H)。

Preparation examples 5 to 6: (R) -tert-butyl [1- (2-methoxyphenyl) -1-oxopropan-2-yl ] carbamate (IX-6)

The same procedures as in preparation examples 5 to 5 were repeated except that 4-methoxybromobenzene was not used but 2-methoxybromobenzene was used to obtain the title compound: 2.31g (75.9%).

1H NMR(400MHz,CDCl3):1.32(d,3H),1.45(s,9H),3.92(s,3H),5.32(m,1H),5.59(br,1H),7.0(m,2H),7.49(m,1H),7.76(m,1H)。

Preparation examples 5 to 7: (S) -tert-butyl (1- (3, 5-difluorophenyl) -1-oxopropan-2-yl) carbamate (IX-7)

The same procedure as in production example 5-5 was repeated except that (R) -tert-butyl (1- (methoxy (methyl) amino) -1-oxopropan-2-yl) carbamate and 4-methoxybromobenzene were not used but (S) -tert-butyl (1- (methoxy (methyl) amino) -1-oxopropan-2-yl) carbamate and 1-bromo-3, 5-difluorobenzene were used, respectively, to obtain the title compound: 1.3g (46%).

1H NMR(400MHz,CDCl3):1.40(d,J＝7.2Hz,3H),1.46(s,9H),5.15～5.19(m,1H),5.41～5.43(m,1H),7.03～7.08(m,1H),7.48～7.50(m,2H)。

Preparation examples 5 to 8: (R) -tert-butyl (1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) carbamate (IX-8)

The same procedures as in preparation examples 5 to 5 were repeated except that 4-methoxybromobenzene was not used but 1-bromo-2, 6-difluorobenzene was used to obtain the title compound: 0.16g (10%).

1H NMR(400MHz,CDCl3):1.39(d,J＝7.2Hz,3H),1.41(s,9H),4.87～5.19(m,1H),5.30～5.31(m,1H),6.94～7.0(m,2H),7.39～7.46(m,1H)。

Preparation examples 5 to 9: (S) -tert-butyl (1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) carbamate (IX-9)

The same procedures as in preparation examples 5 to 5 were repeated except that (S) -tert-butyl (1- (methoxy (methyl) amino) -1-oxopropan-2-yl) carbamate and 1-bromo-2, 6-difluorobenzene were used instead of (R) -tert-butyl (1- (methoxy (methyl) amino) -1-oxopropan-2-yl) carbamate and 4-methoxybromobenzene, respectively, to obtain the title compound: 0.26g (17%).

1H NMR(400MHz,CDCl3):1.39(d,J＝7.2Hz,3H),1.41(s,9H),4.83～4.87(m,1H),5.30～5.31(m,1H),6.94～6.99(m,2H),7.39～7.46(m,1H)。

Preparation examples 5 to 10: (R) -tert-butyl [1- (3-methoxyphenyl) -1-oxopropan-2-yl ] carbamate (IX-10)

The same procedures as in preparation examples 5 to 5 were repeated except that 4-methoxybromobenzene was not used but 3-methoxybromobenzene was used to obtain the title compound: 2.56g (71%).

1H NMR(400MHz,CDCl3):1.32(d,3H),1.45(s,9H),3.94(s,3H),5.32(m,1H),5.61(br,1H),7.12(m,3H),7.62(m,1H)。

Preparation examples 5 to 11: (R) -tert-butyl [1- (hydroxyimino) -1- (3-methoxyphenyl) propan-2-yl ] carbamate (IX-11)

The compound (IX-10) (0.3g,1.1mmol) obtained in production examples 5 to 10, hydroxylamine hydrochloride (0.18g,2.7mmol) and sodium acetate (0.2g,2.7mmol) were dissolved in 15mL of a mixture of ethanol and water (5/1), followed by heating under reflux for 3 hours.

After the reaction was complete, the reaction mixture was cooled to 20-25 ℃, neutralized with 1.3mL of saturated sodium carbonate solution, and then concentrated by vacuum distillation. The concentrate was dissolved in 15mL of water and 20mL of ethyl acetate. The aqueous layer was then extracted three times with 20mL of ethyl acetate. The resulting organic layers were combined, dried over anhydrous magnesium sulfate, and concentrated under vacuum. The concentrate was purified by column chromatography to give the title compound as a mixture of E-and Z-isomers (1: 2): 0.27g (85.2%).

1H NMR (400MHz in CDCl3) 1.39(s,9H),1.43(m,3H),3.78(s,3H),4.69(m,1H),5.26(br,1H), 7.33-6.89 (m,4H) (E-isomer).

1H NMR (400MHz in CDCl3) 1.39(s,9H),1.43(m,3H),3.78(s,3H),5.05(m,1H),5.82(br,1H), 7.33-6.89 (m,4H) (Z-isomer).

Preparation examples 5 to 12: tert-butyl ((2R) -1-amino-1-phenylpropan-2-yl) carbamate (IX-12)

In a reaction vessel which was first evacuated and then purged with argon, (R) -tert-butyl [ 1-phenyl-1-oxopropan-2-yl ] carbamate (0.75g, 3.0mmol) and ammonium chloride (0.24g, 4.5mmol) were placed. Subsequently, dry tetrahydrofuran (15mL) and triethylamine (0.63mL, 4.5mmol) were added to the reaction vessel followed by tetraisopropyl titanate (1.32mL, 4.5 mmol). The reaction mixture is stirred at room temperature for 8 to 10 hours. After the addition of ammonium borane (ammoniumborane), the reaction mixture is stirred again for 8-10 hours.

Extraction was performed with 1M aqueous ammonia (15mL) and diethyl ether (15mL), after which the organic layer was washed with 9mL of 1M HCl. The aqueous layer was adjusted to pH 7-8 with 1M sodium hydroxide and extracted with 15mL of diethyl ether. The resulting organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum to give the title compound: 0.096g (17%).

1H NMR(400MHz,CDCl3):0.95(d,1H),1.02(d,3H),1.20(d,1H),1.46(s,9H),7.26～7.34(m,5H)。

Preparation examples 5 to 13: benzyl tert-butyl ((2R) -1-phenylpropane-1, 2-diyl) dicarbamate (IX-13)

5mL of methylene chloride and 0.45mL (3.23mmol) of triethylamine were added to the compound (IX-12) (0.37g, 1.47mmol) obtained in preparation examples 5-12, followed by slowly adding benzyl chloroformate at 0 ℃. After stirring at room temperature for 12-18 hours, extracting with water and ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give the title compound: 0.37g (49%).

1H NMR(400MHz,CDCl3):1.09(d,3H),1.42(s,9H),3.93～4.42(m,2H),4.70～4.75(m,1H)4.97～5.17(m,2H),6.02～6.33(d,1H),6.92～7.34(m,5H)。

Preparation examples 5 to 14: tert-butyl ((2S) -1-amino-1-phenylpropan-2-yl) carbamate (IX-14)

The same procedures as in preparation examples 5 to 12 were repeated except that (R) -tert-butyl [ 1-phenyl-1-oxopropan-2-yl ] carbamate was not used and (S) -tert-butyl [ 1-phenyl-1-oxopropan-2-yl ] carbamate was used to obtain the title compound: 0.5g (67%).

1H NMR(400MHz,CDCl3):0.95(d,1H),1.01(d,3H),1.20(d,1H),1.45(s,9H),7.24～7.34(m,5H)。

Preparation examples 5 to 15: benzyl tert-butyl ((2S) -1-phenylpropane-1, 2-diyl) dicarbamate (IX-15)

The same procedures as in production examples 5 to 13 were repeated except that the compound (IX-14) obtained in production examples 5 to 14 was used instead of the compound (IX-12) obtained in production examples 5 to 12 to obtain the title compound: 0.22g (28%).

1H NMR(400MHz,CDCl3):0.98(d,3H),1.40(s,9H),3.93～4.41(m,1H),4.68～4.73(m,1H),4.97～5.18(m,2H),6.03～6.33(d,1H),7.21～7.52(m,5H)。

Preparation example 6: preparation of the Compound of formula X

Preparation example 6-1: (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-1)

The compound (IX-1) (0.66g,4.18mmol) in production example 5-1 was added dropwise at 0 ℃ to a solution of the compound (VIII-2) (1.00g,3.48mmol) obtained in production example 4-2 in 10mL of dimethylformamide. Subsequently, diisopropylethylamine (1.73mL,10.44mmol) and benzotriazol-1-yl-oxy-tris (dimethylamino) phosphonium hexafluorophosphate (1.54g,3.48mmol) were added, followed by stirring at 20-25 ℃ for 16 hours.

When the reaction was complete, the reaction solvent was removed by vacuum concentration, and the residue was dissolved in ethyl acetate and extracted twice with water, followed by washing with brine. The organic layer was then dried over anhydrous magnesium sulfate and concentrated in vacuo, followed by purification by column chromatography (ethyl acetate: cyclohexane ═ 1:3 → ethyl acetate: cyclohexane ═ 1:1) to give the title compound as a white oil: 1.31g (85%).

1H NMR(400MHz,CDCl3):1.22(d,3H),1.46(s,9H),1.65～1.86(m,4H),2.33～3.37(m,1H),2.82(t,2H),3.22～3.24(m,1H),3.29(s,3H),3.4(s,3H),3.48～3.55(m,2H),3.75～3.83(m,2H),5.77～6.51(m,1H),6.65～6.74(m,3H)。

Preparation example 6-2: (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2, 4-dichloro-5-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-2)

The same procedure as in preparation example 6-1 was repeated, except that the compound (IX-3) obtained in preparation example 5-3 was used instead of the compound (IX-1) obtained in preparation example 5-1, to obtain the title compound: 0.65g (78%).

1H NMR(400MHz,CDCl3):1.25(d,3H),1.49(s,9H),1.67～1.88(m,4H),2.33～3.35(m,1H),2.94(t,2H),3.23(m,1H),3.24(s,3H),3.41(s,3H),3.46～3.52(m,2H),3.77～3.82(m,2H),5.79～6.58(m,1H),7.10(m,1H),7.40(m,1H)。

Preparation examples 6 to 3: (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((3-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-3)

The same procedure as in preparation example 6-1 was repeated, except that the compound (IX-2) obtained in preparation example 5-2 was used instead of the compound (IX-1) obtained in preparation example 5-1, to obtain the title compound: 0.65g (95%).

1H NMR(400MHz,CDCl3):1.24(m,3H),1.48(s,9H),1.68(m,3H),1.85(m,2H),2.34(m,1H),2.83(t,2H),3.20～3.41(m,1H),3.29(s,3H),3.43(s,3H),3.51～3.56(m,2H),3.66～3.83(m,2H),5.71～6.38(m,1H),6.92(d,1H),6.99(d,1H),7.24(m,2H)。

Preparation examples 6 to 4: (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((4-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-4)

The same procedure as in preparation example 6-1 was repeated except that the compound (IX-1) obtained in preparation example 5-1 was not used but 2- (4-fluorophenyl) ethylamine was used to obtain the title compound: 0.66g (93%).

1H NMR(400MHz,CDCl3):1.24(m,3H),1.49(s,9H),1.65(m,3H),1.76(m,2H),2.32(m,1H),2.80(t,2H),3.22(m,1H),3.23(s,3H),3.27(s,3H),3.43(s,3H),3.48～3.55(m,2H),3.72～3.82(m,2H),5.68～6.32(m,1H),6.97(s,2H),7.16(d,2H)。

Preparation examples 6 to 5: (2R,4R) -tert-butyl-4- ((tert-butyldimethylsilyl) oxy) -2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidine-1-carboxylic acid ester (X-5)

The same procedures as in production example 6-1 were repeated except that the compound (VIII-2) obtained in production example 4-2 and the compound (IX-1) obtained in production example 5-1 were not used, and the compound (VIII-3) obtained in production example 4-3 and 2-phenyethylamine (2-phenyetamine) were used, respectively, to obtain the title compound: 0.71g (95%).

1H NMR(400MHz,CDCl3)0.04(s,6H),0.85(s,9H),1.09(m,3H),1.46(s,9H),1.70～2.82(m,3H),3.26～3.33(m,2H),3.37(s,3H),3.74(m,2H),3.86～4.3(m,4H),5.70～6.25(m,2H),7.21～7.30(m,5H)

Preparation examples 6 to 6: (R) -tert-butyl-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) -4-oxopyrrolidine-1-carboxylic acid ester (X-6)

The same procedure as in production example 6-1 was repeated except that the compounds (VIII-2 and IX-1) obtained in production examples 4-2 and 5-1 were not used but (2R,3R) -3- ((R) -1- (tert-butoxycarbonyl) -4-oxopyrrolidin-2-yl) -3-methoxy-2-methylpropionic acid and 2-cyclohexylethylamine were used, respectively, to obtain the title compound: 0.33g (85%).

1H NMR(400MHz,CDCl3):1.08～1.17(m,3H),1.20～1.27(m,2H),1.57(s,9H),1.73(m,1H),1.99～2.06(m,1H),2.39～2.56(m,2H),2.79～2.88(m,2H),3.34(s,3H),3.47～3.67(m,2H),3.72～3.97(m,3H),4.31～4.41(m,1H),5.67～5.81(m,2H),7.06～7.43(m,5H)。

Preparation examples 6 to 7: (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (2-fluoro-4-methoxyphenyl) -2-oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-7)

The same procedure as in production example 6-1 was repeated except that the compound (IX-4) obtained in production example 5-4 was used instead of the compound (IX-1) obtained in production example 5-1 to obtain the title compound: 0.87g (93%).

1H NMR(400MHz,CDCl3):1.24(m,3H),1.46(s.9H),1.67～1.92(m,4H),2.41～2.45(m.1H),3.19～3.21(m,1H),3.28(s,3H),3.42(s,3H),3.78(m,1H),3.85(s,3H),3.92(m,1H),4.60(s,2H),6.77(m,2H),7.95(m,1H)。

Preparation examples 6 to 8: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2-phenylethyl) amino) propyl) pyrrolidine-1-carboxylic acid ester (X-8)

The same procedure as in preparation example 6-1 was repeated except that the compound (IX-1) obtained in preparation example 5-1 was not used but 2-amino-1-acetophenone was used to obtain the title compound: 1.7g (93%).

1H NMR(400MHz,CDCl3):1.27(d,3H),1.49(s,9H),1.68～1.73(m,1H),1.82～1.95(m,3H),2.5(m,1H),3.21～3.27(m,1H),3.29(s,3H),3.38～3.65(m,1H),3.48(s,3H),3.82～3.99(m,2H),4.7(S,2H),7.5(m,2H),7.61(m,1H),7.97(d,1H)。

Preparation examples 6 to 9: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2- (p-tolyl) ethyl) amino) propyl) pyrrolidine-1-carboxylic acid ester (X-9)

The same procedure as in preparation example 6-1 was repeated except that the compound (IX-1) obtained in preparation example 5-1 was not used but 2-amino-1- (p-tolyl) ethanone was used to obtain the title compound: 0.33g (81%).

1H NMR(400MHz,CDCl3):1.25(m,3H),1.47(s.9H),1.71～1.95(m,4H),2.43(s.3H),2.47～2.55(m.1H),3.21～3.27(m,1H),3.39～3.42(m,1H),3.48(s,3H),3.81(m,1H),3.92～3.96(m,1H),4.73(s,2H),7.30(d,J＝8.0Hz,2H),7.88(d,J＝8.0Hz,2H)。

Preparation examples 6 to 10: (2S,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (4-fluorophenyl) -2-oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-10)

The same procedure as in preparation example 6-1 was repeated except that the compound (IX-1) obtained in preparation example 5-1 was not used but 2-amino-1- (4-fluorophenyl) ethanone was used to obtain the title compound: 0.56g (81%).

1H NMR(400MHz,CDCl3):1.26～1.33(m,3H),1.49(s,9H),1.69～1.95(m,4H),2.48～2.56(m,1H),3.21～3.27(m,1H),3.29(s,3H),3.43～3.47(m,1H),3.48(s,3H),3.80～3.82(m,1H),3.92～3.98(m,1H),4.73(s,2H),7.61(m,2H),8.01～8.02(m,2H)。

Preparation 6-11 (2R,4S) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidine-1-carboxylic acid ester (X-11)

The same procedure as in production example 6-1 was repeated except that the compounds (VIII-2 and IX-1) obtained in production examples 4-2 and 5-1 were not used but (2R,3R) -3- ((2R,4S) -1- (tert-butoxycarbonyl) -4-methoxypyrrolidin-2-yl) -3-methoxy-2-methylpropanoic acid and 2-cyclohexylethylamine were used, respectively, to obtain the title compound: 0.26g (61%).

1H NMR(400MHz,CDCl3):1.18(d,3H),1.47(s,9H),1.94～2.01(m,4H),2.26～2.38(m,1H),2.83(t,2H),2.98(m,1H),3.32(s,3H),3.39(s,3H),3.48～3.52(m,2H),3.73～3.79(m,2H),5.67～6.07(m,1H),6.99～7.32(m,4H)。

Preparation examples 6 to 12: (2S,4R) -tert-butyl-2- ((1R,2R) -3- ((2-hydroxy-2-phenylethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-12)

The same procedure as in production example 6-1 was repeated except that the compound (IX-1) obtained in production example 5-1 was not used but 2-amino-1-phenylethyl alcohol was used to obtain the title compound: 0.43g (100%).

1H NMR(400MHz,CDCl3):1.23～1.31(m,3H),1.49(s,9H),1.73～1.99(m,4H),2.34～2.41(m,1H),3.06～3.43(m,3H),3.29(s,3H),3.45(s,3H),3.52～4.84(m,4H),4.33～4.99(m,2H),7.28～7.45(m,5H)。

Preparation examples 6 to 13: (2S,4R) -tert-butyl-2- (6R,9R,10R) -6, 9-dimethyl-3, 8-dioxo-1, 5-diphenyl-2, 11-dioxa-4, 7-diazacyclo-n-10-yl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-13)

The same procedure as in production example 6-1 was repeated except that the compound (IX-13) obtained in production example 5-13 was used instead of the compound (IX-1) obtained in production example 5-1 to obtain the title compound: 68mg (39%).

1H NMR(400MHz,CDCl3):0.98～1.28(m,7H),1.43(s,9H),1.71～2.29(m,4H),3.21～3.30(m,2H),3.27(s,3H),3.38(s,3H),3.72～3.75(m,2H),4.33～4.69(m,1H),5.0(s,2H),5.82～6.13(m,1H),6.46～7.0(m,1H),7.21～7.40(m,10H)。

Preparation examples 6 to 14: (2S,4R) -tert-butyl 2- (6S,9R,10R) -6, 9-dimethyl-3, 8-dioxo-1, 5-diphenyl-2, 11-dioxa-4, 7-diazidon-10-yl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-14)

The same procedure as in production example 6-1 was repeated, except that the compound (IX-15) obtained in production example 5-15 was used instead of the compound (IX-1) obtained in production example 5-1, to obtain the title compound: 0.13g (73%).

1H NMR(400MHz,CDCl3):0.98～1.26(m,7H),1.40(s,9H),1.71～2.29(m,4H),3.20～3.26(m,2H),3.29(s,3H),3.37(s,3H),3.46～3.72(m,2H),4.28～4.43(m,1H),5.0(s,2H),5.82～6.13(m,1H),6.46～7.0(m,1H),7.21～7.40(m,10H)。

Preparation examples 6 to 15: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2- (pyridin-2-yl) ethyl) amino) propyl) pyrrolidine-1-carboxylic acid ester (X-15)

The same procedure as in production example 6-1 was repeated except that the compound (IX-1) obtained in production example 5-1 was not used but 2- (pyridin-2-yl) ethylamine was used to obtain the title compound: 0.5g (64%).

1H NMR(400MHz,CDCl3):1.20(s,3H),1.47(d,J＝6.8Hz,9H),1.71(m,2H),1.72(m,2H),1.99(s,1.H),2.31(m,1H),3.00(t,2H),3.21(t,1H),3.27(s,3H),3.39(s,3H),3.53～3.84(m,4H),7.16(t,2H),7.61(t,1H),8.52(m,1H)。

Preparation examples 6 to 16: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2- (thiophen-2-yl) ethyl) amino) propyl) pyrrolidine-1-carboxylic acid ester (X-16)

The same procedure as in production example 6-1 was repeated except that the compound (IX-1) obtained in production example 5-1 was not used but 2- (thiophen-2-yl) ethylamine was used to obtain the title compound: 0.8g (99%).

1H NMR(400MHz,CDCl3):1.21(m,3H),1.48(d,J＝12.8Hz,9H),1.74(m,2H),1.88(m,2H),2.28～2.38(m,1H),3.05(t,J＝6.4Hz,2H),3.21(m,1H),3.25(s,3H),3.39(s,3H),3.53～3.57(m,2H),3.74～3.86(m,2H),6.85(s,1H),6.94(t,J＝3.6Hz,1H),7.15(s,1H)。

Preparation examples 6 to 17: (2S,4R) -tert-butyl-2- ((1R,2R) -3- (((S) -1- (3, 5-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-17)

4mL of trifluoroacetic acid was added dropwise to a solution of the compound (IX-7) (0.36g,1.26mmol) obtained in production examples 5-7 in 5mL of dichloromethane while stirring at 20 to 25 ℃ for 3 hours. After completion of the reaction, the reaction solvent was removed by vacuum concentration. After toluene was added twice in an amount of 5mL to completely remove trifluoroacetic acid, the reaction was continued.

The reaction concentrate (TFA salt) and the compound (VIII-2) (0.40g,1.26mmol) obtained in preparation example 4-2 were dissolved in 5mL of dimethylformamide. Diethyl cyanophosphate (DEPC) (0.19mL,1.32mmol) and triethylamine (0.89mL,6.30mmol) were added to the solution at 0 deg.C, followed by stirring at 20-25 deg.C for 16 hours. When the reaction was complete, the reaction solvent was removed by vacuum concentration. The residue was dissolved in ethyl acetate and extracted with 1M potassium bisulfite, water, saturated sodium bicarbonate solution and brine, and the organic layer thus obtained was dried over anhydrous sodium sulfate and concentrated in vacuo. The concentrate was purified by column chromatography (ethyl acetate: cyclohexane ═ 2:1 → ethyl acetate) to give the title compound: 0.53g (94%).

1H NMR(400MHz,CDCl3):1.19(m,3H),1.38(s.9H),1.52(m,3H),1.85～1.91(m,5H),2.92～2.94(m.2H),3.30(s,6H),3.61～3.67(m,4H),5.27(m,1H),6.86(m,1H),7.32(m,2H),8.03(br,1H)。

Preparation examples 6 to 18: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (2-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidine-1-carboxylic acid ester (X-18)

The same procedures as in production examples 6 to 17 were repeated except that the compound (IX-6) obtained in production examples 5 to 6 was used instead of the compound (IX-7) obtained in production examples 5 to 7 to obtain the title compound: 0.21g (75%).

1H NMR(400MHz,CDCl3):1.26(m,3H),1.34(m,3H),1.49(s,9H),1.71～2.45(m,4H),3.25(s.3H),3.33(m,1H),3.47(s,3H),3.71～3.89(m,2H),3.94(s,3H),4.05(m,1H),5.55(m,1H),6.82(d,1H),7.03(m,2H),7.52(m,1H),7.76(d,1H)

Preparation examples 6 to 19: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (3-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidine-1-carboxylic acid ester (X-19)

The same procedures as in preparation examples 6 to 17 were repeated except that the compound (IX-7) obtained in preparation examples 5 to 7 was not used and (R) -tert-butyl (1- (3-methoxyphenyl) -1-oxopropan-2-yl) carbamate was used to obtain the title compound: 0.18g (64%).

1H NMR(400MHz,CDCl3):1.26(m,3H),1.34(m,3H),1.49(s,9H),1.71～2.45(m,4H),3.22(s.3H),3.33(m,1H),3.47(s,3H),3.71～3.80(m,2H),3.83(s,3H),4.05(m,1H),5.55(m,1H),6.77(d,1H),7.16(m,1H)7.40(m,1H),7.49(m,1H),7.58(d,1H)。

Preparation examples 6 to 20: (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (4-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidine-1-carboxylic acid ester (X-20)

The same procedures as in production examples 6 to 17 were repeated except that the compound (IX-5) obtained in production examples 5 to 5 was used instead of the compound (IX-7) obtained in production examples 5 to 7 to obtain the title compound: 0.23g (82%).

1H NMR(400MHz,CDCl3):1.26(m,3H),1.34(m,3H),1.49(s,9H),1.71～2.45(m,4H),3.21(s.3H),3.33(m,1H),3.46(s,3H),3.71～3.80(m,2H),3.86(s,3H),4.05(m,1H),5.35(m,1H),6.79(d,1H),6.98(m,2H),7.98(m,2H)。

Preparation examples 6 to 21: (2S,4R) -tert-butyl-2- ((1R,2R) -3- (((R) -1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-21)

The same procedures as in production examples 6 to 17 were repeated except that the compound (IX-8) obtained in production examples 5 to 8 was used instead of the compound (IX-7) obtained in production examples 5 to 7 to obtain the title compound: 0.13g (46%).

1H NMR(400MHz,CDCl3):1.26(m,3H),1.34(m,3H),1.49(s,9H),1.71～2.45(m,4H),3.21(s.3H),3.33(m,1H),3.46(s,3H),3.71～3.80(m,2H),4.05(m,1H),5.35(m,1H),6.79(d,1H),7.09(m,2H),7.46(m,1H)。

Preparation examples 6 to 22: (2S,4R) -tert-butyl-2- ((1R,2R) -3- (((S) -1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-22)

The same procedures as in production examples 6 to 17 were repeated except that the compound (IX-9) obtained in production examples 5 to 9 was used instead of the compound (IX-7) obtained in production examples 5 to 7 to obtain the title compound: 0.13g (46%).

Preparation examples 6 to 23: (2S,4R) -tert-butyl-2- ((1R,2R) -3- (((R, E) -1- (hydroxyimino) -1- (3-methoxyphenyl) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylic acid ester (X-23)

The same procedures as in production examples 6 to 17 were repeated except that the compound (IX-11) obtained in production examples 5 to 11 was used instead of the compound (IX-7) obtained in production examples 5 to 7 to obtain the title compound: 0.23g (52%).

1H NMR(400MHz,CDCl3):1.28(m,6H),1.45(m,9H),1.75(m,1H),1.95(m,1H),2.50(m,1H),3.25(m,1H),3.30(s,3H),3.45(s,3H),3.77(s,3H),3.80-3.98(m,2H),5.05(m,1H),5.83(m,1H),6.90(m,2H),7.15(m,1H),7.30(m.1H)。

Preparation examples 6 to 24: (S) -tert-butyl-4- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) thiazolidine-3-carboxylate (X-24)

The same procedure as in preparation example 6-1 was repeated, except that the compound (VIII-2) obtained in preparation example 4-2 was not used, but the compound (VIII-4) obtained in preparation example 4-4 was used, to obtain the title compound: 0.32g (64%).

1H NMR(400MHz,CDCl3):1.17～1.25(d,3H),1.47(s,9H),2.65～3.12(m,8H),3.42(S,3H),3.77～4.18(m,2H),4.16～4.21(m,2H),5.11(br,1H),7.32(m,2H),7.59(m,1H)。

Preparation examples 6 to 25: (2S,4S) -tert-butyl-2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidine-1-carboxylic acid ester (X-25)

The same procedure as in preparation 6-1 was repeated, except that the compound (VIII-2) obtained in preparation 4-2 was not used, but the compound (VIII-4) obtained in preparation 4-4 was used, to obtain the title compound: 3.5g (75%).

1H NMR(400MHz,CDCl3):1.26(m,3H),1.48(d,9H),1.62～1.66(m,2H),1.81～1.89(m,1H),2.07～2.14(m,1H),2.22～2.36(m,1H),2.87～2.92(m,2H),3.36～3.41(m,2H),3.52(s,6H),3.81～3.91(m,1H),3.98～4.00(m,1H),4.11～4.13(m,1H),4.62～4.64(d,1H),5.59(brs,1H),6.03(brs,1H),6.87(m,2H),7.18～7.19(m,1H)。

Preparation examples 6 to 26: (2S,4S) -tert-butyl-2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidine-1-carboxylic acid ester (X-26)

The same procedures as in production examples 6 to 17 were repeated except that the compound (VIII-4) obtained in production examples 4-4 and (S) -tert-butyl (1- (4-fluorophenyl) -1-oxopropan-2-yl) carbamate used in production examples 4-4 and 5-7 were respectively used instead of the compounds (VIII-2 and IX-7) obtained in production examples 4-2 and 5-7 to obtain the title compound: 0.25g (81%).

LC-MS m/z:[M+]+。

Preparation examples 6 to 27: (2S,4S) -tert-butyl-4-azido-2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) pyrrolidine-1-carboxylic acid ester (X-27)

The same procedure as in preparation 6-1 was repeated, except that the compound (VIII-2) obtained in preparation 4-2 was not used, but the compound (VIII-5) obtained in preparation 4-5 was used, to obtain the title compound: 5.9g (94%).

[α]D 25＝-17.35°(c＝1,MeOH)。

1H NMR(400MHz,CDCl3):1.18(d,3H),1.47(s,9H),1.97～2.00(m,1H),2.05～2.39(m,2H),2.90～2.93(t,2H),2.95～2.99(m,1H),3.43(s,3H),3.48～3.53(m,2H),3.75～4.06(m,4H),5.73(brs,1H),6.08(brs,1H),6.87(t,2H,J＝7.6Hz,15.2Hz),7.15～7.21(m,1H)。

Preparation examples 6 to 28: (2S,4S) -tert-butyl-4- (benzyl (methyl) amino) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) pyrrolidine-1-carboxylic acid ester (X-28)

A solution of compound (X-27) (5.37g,11.5mmol) obtained in production examples 6 to 27 in 100mL of methanol was stirred in the presence of 10% palladium on carbon (0.5g) under a hydrogen atmosphere for 16 hours. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was washed several times with methanol. The solvent was removed under vacuum to give (2S,4S) -tert-butyl-4-amino-2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) pyrrolidine-1-carboxylic acid ester 4.96g (98%).

[α]D 25＝-39.65°(c＝1,MeOH)

1H NMR(400MHz,CDCl3):1.17(m,3H),1.47(m,9H),1.88(m,2H),2.94～2.00(m,1H),2.10(s,3H),2.23～2.45(m,1H),2.76～2.78(m,1H),2.89～2.92(m,2H),2.98～3.03(m,1H),3.44～3.55(m,1H),3.47(s,3H),3.78～4.05(m,3H),5.80(brs,1H),6.28(brs,1H),6.84(t,2H,J＝7.6Hz,15.2Hz),7.12～7.33(m,7H)。

The residue was dissolved in 100mL of methanol under an argon atmosphere. Benzaldehyde (1.07ml,10.6mmol) was added to the solution and stirred at 20-25 ℃ for 1 hour. Sodium borocyanide (0.67g,10.6mmol) was added to the solution and then stirred for 16 hours. Again, the resulting solution was mixed with paraformaldehyde (0.91g,10.1mmol) and stirred for 2 hours, followed by the addition of sodium borocyanide (0.67g,10.6mmol) and stirring for 4 hours. When the reaction was complete, the methanol was removed by concentration in vacuo, and the residue was dissolved in 100mL of ethyl acetate and washed with saturated sodium bicarbonate solution. The resulting aqueous layer was extracted twice with ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate and concentrated to dryness under vacuum. The residue was purified by column chromatography to give the title compound: 2.74g (50%).

LC-MS m/z:545.7[M+H]+。

1H NMR(400MHz,CDCl3):1.19(m,3H),1.51(m,9H),1.88(m,2H),2.94～2.00(m,1H),2.21(s,3H),2.27～2.41(m,1H),2.76～2.78(m,1H),2.89～2.92(m,2H),2.98～3.03(m,1H),3.44～3.55(m,1H),3.47(s,3H),3.78～4.05(m,3H),5.8(brs,1H),6.28(brs,1H),7.12～7.19(m,1H),7.22～7.25(m,1H)。

Example (b): preparation of the Compound of formula I

Example 1: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-1)

2mL of trifluoroacetic acid was added dropwise to a solution of the compound obtained in preparation example 6-1 in 3mL of dichloromethane, followed by stirring at 20-25 ℃ for 3 hours. After the reaction was complete, the reaction solvent was removed by vacuum concentration. After toluene was added twice in an amount of 5mL to completely remove trifluoroacetic acid, the reaction was continued.

The reaction concentrate (TFA salt) and (5S,8S,11S,12R) -11- ((S) -sec-butyl) -5, 8-diisopropyl-12-methoxy-4, 10-dimethyl-3, 6, 9-trioxo-1-phenyl-2-oxa-4, 7, 10-triazatetradecane-14-oic acid (compound II-1) (1.00g,2.04mmol) were dissolved in 5mL of dimethylformamide. Diethyl cyanophosphate (DEPC) (0.34mL,2.05mmol) and triethylamine (1.44mL,10.22mmol) were added to the solution at 0 deg.C, followed by stirring at room temperature for 16 hours. When the reaction was complete, the reaction solvent was removed by vacuum concentration. The residue was dissolved in 20mL of ethyl acetate and extracted with 1M potassium bisulfite, water, saturated sodium bicarbonate solution and brine, and the organic layer thus obtained was dried over anhydrous sodium sulfate and concentrated in vacuo. The concentrate was purified by column chromatography (ethyl acetate: cyclohexane ═ 2:1 → ethyl acetate) to give the compound (I-1) whose N-terminal amino group was protected as a pale yellow oil: 1.29g (74%).

1H NMR(400MHz,CDCl3):0.75～0.98(m,19H),1.06(m,1H),1.18～1.27(m,4H),1.38(m,1H)1.91～2.04(m,4H),2.21～2.52(m,4H),2.87～2.91(m,5H),2.99(m,2H),3.1(s,1H),3.31(s,6H),3.37(s,3H),3.43～3.51(m,3H),3.82(dd,1H),4.09～4.15(m,2H),4.68(m,1H),5.12～5.23(m,2H),6.52(m,1H),6.82～6.87(m,2H),7.13～7.17(m,1H),7.30～7.34(m,5H)。

The compound (1.29g,1.47mmol) in which the N-terminal amino group of the compound (I-1) was protected was dissolved in 9mL of t-butyl alcohol and 1mL of water, and reacted in the presence of 10% palladium on carbon (0.1g) under a hydrogen atmosphere for 3 hours while stirring. After completion of the reaction, the reaction mixture was filtered through celite, and washed several times with methanol. The solvent was removed under vacuum to give the title compound as a pale yellow solid: 1.06g (100%).

1H NMR(400MHz,CDCl3):0.78～0.98(m,20H),1.06(m,1H),1.16～1.27(m,4H),1.38(m,1H)1.65～1.76(m,7H),1.89～2.02(m,5H),2.31～2.38(m,5H),2.71～2.75(m,1H),2.88～2.89(m,2H)3.00(s,2H),3.10(d,1H),3.29(s,6H),3.35(s,3H),3.41～3.48(m,3H),3.79～3.82(dd,1H),4.08～4.15(m,2H),4.72～4.86(m,2H),6.56(m,1H),6.79～6.85(m,2H),7.10～7.15(m,1H),7.16～7.22(m,1H),7.55(d,1H)。

Example 2: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((3-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-2)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-3) obtained in preparation example 6-3 was used, to obtain an N-terminal amino-protected compound of the compound (I-2): 0.58g (84%).

1H NMR(400MHz,CDCl3):0.78～0.99(m,19H),1.06(m,1H),1.19～1.25(m,4H),1.34(m,1H)1.67～1.74(m,4H),2.16～2.38(m,4H),2.83～2.91(m,5H),3.00～3.12(m,2H)3.10(s,1H),3.29(s,3H),3.31(s,3H),3.38(s,3H),3.44～3.51(m,3H),3.82(dd,1H),4.09～4.15(m,2H),4.78(m,1H),5.19～5.22(m,2H),6.52(m,1H),6.89～6.98(m,3H),7.23(m,1H),7.31～7.34(m,5H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-2): 0.48g (100%).

1H NMR(400MHz,CDCl3):0.81～0.98(m,20H),1.06(m,1H),1.19～1.25(m,4H),1.38(m,1H)1.65～1.79(m,7H),1.90～2.06(m,5H),2.31～2.41(m,5H),2.71～2.75(m,1H),2.88～2.89(m,2H)3.03(s,2H),3.10(d,1H),3.31(s,6H),3.37(s,3H),3.4～3.56(m,3H),3.82～3.85(dd,1H),4.02～4.13(m,2H),4.75～4.79(m,2H),6.64(m,1H),6.93～7.02(m,2H),7.12～7.18(m,2H)。

Example 3: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((4-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-3)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-4) obtained in preparation example 6-4 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-3): 1.57g (92%).

1H NMR(400MHz,CDCl3):0.78～0.99(m,19H),1.06(m,1H),1.19～1.27(m,4H),1.38(m,1H)1.91～2.04(m,4H),2.17～2.38(m,4H),2.77～2.91(m,5H),3.00～3.12(m,2H)3.10(s,1H),3.29(s,3H),3.31(s,3H),3.38(s,3H),3.44～3.51(m,3H),3.82(dd,1H),4.09～4.15(m,2H),4.68(m,1H),5.12～5.22(m,2H),6.52(m,1H),6.93～6.98(m,3H),7.14～7.18(m,3H),7.31～7.34(m,6H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-3): 0.60g (100%).

1H NMR(400MHz,CDCl3):0.81～0.98(m,20H),1.06(m,1H),1.19～1.25(m,4H),1.38(m,1H)1.65～1.79(m,7H),1.90～2.06(m,5H),2.31～2.41(m,5H),2.71～2.75(m,1H),2.88～2.89(m,2H)3.03(s,2H),3.10(d,1H),3.31(s,6H),3.37(s,3H),3.41～3.56(m,3H),3.82～3.85(dd,1H),4.02～4.13(m,2H),4.75～4.79(m,2H),6.64(m,1H),6.86～7.00(m,3H),7.57～7.60(m,1H)。

Example 4: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2, 4-dichloro-5-fluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-4)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-2) obtained in preparation example 6-2 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-4): 0.94g (75%).

1H NMR(400MHz,CDCl3):0.68～0.97(m,19H),1.07(m,1H),1.18～1.25(m,4H),1.34(m,1H)1.70～1.78(m,4H),1.91～2.04(m,3H),2.21～2.42(m,3H),2.87～2.91(m,5H),3.27(s,3H),3.31(s,3H),3.38(s,3H),3.44～3.51(m,3H),3.82(dd,1H),4.09～4.15(m,2H),4.66～4.70(m,1H),5.09～5.22(m,2H),6.52(m,1H),6.87(m,1H),7.09～7.11(m,1H),7.30～7.34(m,5H),7.40～7.44(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-4): 0.70g (92%).

1H NMR(400MHz,CDCl3):0.71～0.99(m,19H),1.05(m,1H),1.18～1.25(m,4H),1.34(m,1H)1.70～1.78(m,4H),1.21～2.04(m,3H),2.21～2.42(m,3H),2.87～2.91(m,5H),3.31(s,6H),3.38(s,3H),3.44～3.51(m,3H),3.82(dd,1H),4.09～4.15(m,2H),4.68～4.72(m,1H),5.09～5.22(m,2H),6.52(m,1H),6.90(m,1H),7.09～7.11(m,1H),7.30～7.34(m,5H),7.40～7.44(m,1H)。

Example 5: (2S) -N- ((3R,4S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- ((2- (4-methoxyphenyl) -2-oxoethyl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-5)

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used but (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-3- ((2- (4-methoxyphenyl) -2-oxoethyl) amino) -2-methyl-3-oxopropyl) pyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of the compound (I-5): 0.35g (78%).

1H NMR(400MHz,CDCl3):0.71～0.88(m,12H),0.95～1.07(m,1H),0.96(d,J＝6.8Hz,3H),1.26～1.33(m,1H),1.32(d,J＝6.8Hz,3H),1.62(s,3H),1.67～1.86(m,2H),2.00～2.08(m,3H),2.24～2.32(m,2H),2.41～2.47(m,2H),2.59(t,J＝7.2Hz,1H),2.87～2.99(m,4H),3.08(br,1H),3.24(s,3H),3.28～3.55(m,2H),3.33(s,1H),3.46(s,3H),3.89(s,3H),3.99～4.11(m,2H),4.13～4.15(m,2H),4.24(m,1H),4.67～4.71(m,4H),5.09～5.23(m,3H),6.49(d,J＝9.2Hz,2H),6.96(d,J＝8.8Hz,2H),7.09(br,1H),7.21～7.34(m,5H),7.95(d,J＝8.8Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-5) was used: 0.26g (90%).

MALDI-TOF MS m/z:731.8[M+H]+。

1H NMR(400MHz,CDCl3):0.79～0.84(m,3H),0.90～1.03(m,12H),1.25～1.33(m,3H),1.68～1.89(m,6H),2.02～2.05(m,3H),2.33(s,3H),2.38～2.61(m,2H),2.74(d,J＝5.2Hz,1H),3.02～3.03(m,3H),3.29(s,3H),3.32～3.34(m,3H),3.40～3.46(m,6H),3.78(s,3H),3.89(s,2H),3.99～4.16(m,3H),4.25(br,1H),4.70～4.78(m,4H),6.70(m,1H),6.96(d,J＝8.8Hz,2H),7.10(br,1H),7.60(m,2H),7.95(d,J＝8.8Hz,2H)。

Example 6: (2S) -N- ((3R,4S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (((R) -1-oxo-1-phenylpropan-2-yl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-6)

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used but (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (((R) -1-oxo-1-phenylpropan-2-yl) amino) propyl) pyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of compound (I-6): 0.27g (88%).

ES-MS m/z:850[M+H]+。

1H NMR(400MHz,CDCl3):0.77～0.90(m,12H),0.96(d,J＝7.2Hz,3H),1.29(d,J＝7.2Hz,3H),1.42(d,J＝7.2Hz,3H),1.73～1.81(m,5H),1.97～2.00(m,3H),2.20～2.33(m,3H),2.42～2.53(m,2H),2.90(s,3H),2.99(s,6H),3.28(s,1H),3.33(s,1H),3.37～3.50(m,2H),3.43(s,3H),3.97(dd,J＝2.8,7.2Hz,1H),4.12～4.15(m,3H),4.61～4.70(m,2H),5.09～5.21(m,3H),5.56(t,J＝7.2Hz,1H),6.51(br,J＝8.8Hz,1H),7.08(br,J＝7.2Hz,1H),7.29～7.35(m,5H),7.49(t,J＝7.2Hz,2H),7.60(t,J＝7.2Hz,1H),7.99(d,J＝7.2Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-6): 0.22g (98%).

MALDI-TOF MS m/z:715.7[M+H]+,738.1[M+Na]+。

1H NMR(400MHz,CDCl3):0.81～1.04(m,9H),0.94(d,J＝6.8Hz,3H),0.99(d,J＝6.8Hz,3H),1.18(d,J＝7.2Hz,3H),1.20(d,J＝7.2Hz,3H),1.34～1.42(m,1H),1.68～1.86(m,5H),1.91～2.09(m,5H),2.31～2.49(m,4H),2.33(s,3H),2.74(m,1H),3.04(s,3H),3.31(s,6H),3.36(s,3H),3.41(s,1H),3.43～3.53(m,1H),3.73～3.93(m,2H),4.08～4.19(m,2H),4.65(d,J＝6.0Hz,1H),4.73～4.86(m,2H),6.92(br,J＝7.6Hz,1H),7.21～7.40(m,5H),7.59～7.62(m,1H)。

Example 7: (2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (((S) -1-oxo-1-phenylpropan-2-yl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-7)

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used but (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (((S) -1-oxo-1-phenylpropan-2-yl) amino) propyl) pyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of compound (I-7): 0.31g (90%).

ES-MS m/z:850[M+H]+。

1H NMR(400MHz,CDCl3):0.78～0.91(m,12H),0.97(d,J＝6.8Hz,3H),1.29(d,J＝6.8Hz,3H),1.43(d,J＝7.2Hz,3H),1.73～1.82(m,5H),1.97～2.06(m,3H),2.23～2.43(m,4H),2.49(t,J＝7.2Hz,1H),2.90(s,3H),2.99(s,6H),3.14(s,1H),3.23～3.53(m,2H),3.34(s,3H),3.43(s,3H),4.00(dd,J＝2.4,7.2Hz,1H),4.13～4.16(m,2H),4.27～4.29(m,1H),4.67～4.81(m,2H),5.10～5.23(m,3H),5.52(t,J＝7.2Hz,1H),6.51(br,J＝8.8Hz,1H),7.27～7.35(m,5H),7.48(t,J＝7.2Hz,2H),7.51～7.61(m,1H),7.59(t,J＝7.2Hz,1H),7.98(d,J＝7.2Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-7): 0.24g (93%).

MALDI-TOF MS m/z:716.0[M+H]+,738.0[M+Na]+。

1H NMR(400MHz,CDCl3):0.82～1.01(m,12H),1.14(d,J＝6.8Hz,3H),1.26(d,J＝6.8Hz,3H),1.30(d,J＝7.2Hz,3H),1.98～2.12(m,3H),2.33(s,3H),2.34(s,3H),2.36～2.50(m,2H),2.73～2.76(m,2H),3.01～3.04(m,3H),3.32～3.54(m,2H),3.30(s,6H),3.39(s,3H),3.45(s,3H),3.81～3.85(m,2H),4.07～4.08(m,2H),4.32～4.48(m,2H),4.58～4.60(m,1H),4.73～4.79(m,2H),6.35(br,J＝8.8Hz,1H),6.64(br,J＝8.0Hz,1H),7.21～7.44(m,6H),7.56～7.60(m,1H)。

Example 8: (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -4-hydroxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-8)

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but the compound (X-5) and (3R,4S,5S) -4- ((S) -2- (dimethylamino) -3-methylbutanamido) -N, 3-dimethylbutyrylamino) -3-methoxy-5-methylheptanoic acid (compound II-2) obtained in preparation example 6-5 were used, respectively, to obtain a hydroxy-protected compound at the compound (I-8): 0.36g (68%).

MALDI-TOF MS m/z:832.9[M+H]+。

1H NMR(400MHz,CDCl3):0.04(s,6H),0.92～1.05(m,20H),1.09(m,3H),1.46(m,4H),1.62(s,2H),1.97～2.17(m,3H),2.44～2.46(m,3H),2.71～2.80(m,3H),2.81～2.89(m,1H),2.91～2.94(m,2H),2.71～2.80(m,3H)3.04(s,3H),3.05(s,3H),3.26～3.33(m,2H),3.37(s,3H),3.74(m,2H),3.86～4.30(m,4H),5.70～6.25(m,2H),7.21～7.30(m,5H)。

The hydroxy-protected compound (0.07g,0.08mmol) of the compound (I-8) was dissolved in 6mL of tetrahydrofuran and reacted with 1.0M tetrabutylammonium fluoride (0.3mL,0.25mmol) with stirring for 5 hours. The reaction was quenched by saturated ammonium chloride solution and the reaction mixture was extracted with 30mL of ethyl acetate and 20mL of water, followed by concentration in vacuo. The residue was purified by column chromatography (dichloromethane: methanol ═ 9:1) to give the title compound: 54mg (90%).

1H NMR(400MHz,CDCl3):0.92～1.05(m,20H),1.09(m,3H),1.46(m,4H),1.62(s,2H),1.97～2.17(m,3H),2.44～2.46(m,3H),2.71～2.80(m,3H),2.81～2.89(m,1H),2.91～2.94(m,2H),2.71～2.80(m,3H)3.04(s,3H),3.05(s,3H),3.26～3.33(m,2H),3.37(s,3H),3.74(m,2H),3.86～4.3(m,4H),5.70～6.25(m,2H),7.21～7.30(m,5H)。

Example 9: (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -3-methoxy-1- ((R) -2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) -4-oxopyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-9)

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in production example 6-1 were not used, but the compound (X-6) and the compound (II-2) obtained in production example 6-6 were used, respectively, to obtain the title compound: 0.25g (72%).

1H NMR(400MHz,CDCl3):0.80～0.88(m,3H),0.92～1.05(m,15H),1.09～1.25(m,4H),1.60(m,3H),1.99～2.17(m,4H),2.24～2.37(m,6H),2.43～2.50(m,5H),2.81～2.84(m,2H),3.04(s,3H),3.28(s,3H),3.38(s,3H),3.53(m,3H),3.73～4.3(m,4H),4.71～4.79(m,2H),5.70(m,1H),7.18～7.36(m,5H)。

EXAMPLE 10 (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4S) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-10)

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in production example 6-1 were not used, and the compound (X-11) and the compound (II-2) obtained in production example 6-11 were used, respectively, to obtain the title compound: 0.80g (73%).

1H NMR(400MHz,CDCl3):0.80～0.83(m,3H),0.92～1.02(m,16H),1.18～1.22(m,4H),1.83(m,3H),2.07～2.17(m,4H),2.24～2.37(m,6H),2.43～2.50(m,4H),2.81～2.88(m,2H),3.02(s,3H),3.27(s,3H),3.38(s,3H),3.49～3.53(m,3H),3.87～4.22(m,4H),4.76～4.79(m,1H),6.40(m,1H),7.18～7.38(m,5H)。

Example 11: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-11)

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used but (2R,4R) -tert-butyl-2- ((1R,2R) -3- (((R) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of the compound (I-11): 0.55g (68%).

1H NMR(400MHz,CDCl3):0.71～1.03(m,20H),1.22(d,3H),1.33～1.37(m,4H),1.40～1.44(m,3H),1.60～1.81(m,4H),1.98(m,2H),2.26(m,2H),2.90(s,3H),3.21(s,3H),3.32(s,3H),3.43(s,3H),4.06～4.15(m,2H),4.22～4.27(m,2H),4.64～4.69(m,2H),5.09～5.13(m,2H),5.23～5.30(m,2H),5.50(m,1H),7.18(t,2H),7.34(m,5H),8.01～8.05(m,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-11) was used: 0.44g (100%).

1H NMR(400MHz,CDCl3):0.71～1.03(m,20H),1.22(d,3H),1.33～1.37(m,4H),1.40～1.44(m,3H),1.60～1.81(m,4H),1.98(m,2H),2.26(m,2H),2.90(s,3H),3.21(s,3H),3.32(s,3H),3.43(s,3H),4.06～4.15(m,2H),4.22～4.27(m,2H),4.64～4.69(m,2H),5.09～5.13(m,2H),5.23～5.30(m,2H),5.50(m,1H),7.18(t,2H),8.01～8.05(m,2H)。

Example 12: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-12)

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used, but (2R,4R) -tert-butyl-2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of the compound (I-12): 0.54g (67%).

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-12): 0.45g (100%).

Example 13: (S) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (2-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-13)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-18) obtained in preparation example 6-18 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-13): 0.23g (69%).

1H NMR(400MHz,CDCl3):0.78～0.99(m,19H),1.29～1.42(m,6H),1.69(m,6H),1.95～2.04(m,2H),2.21～2.43(m,3H),2.55(m,1H),2.90～3.00(m,6H),3.21～3.42(m,7H),3.55(m,1H),3.86(m,2H),4.01～4.26(m,3H),4.66～4.69(m,2H),5.12～5.22(m,2H),5.52(m,1H),6.54(d,1H),7.02(m,2H),7.34(m,5H),7.52(m,1H),7.75(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-13) was used: 0.16g (100%).

1H NMR(400MHz,CDCl3):0.78～0.99(m,19H),1.29～1.42(m,6H),1.69(m,6H),1.95～2.04(m,2H),2.21～2.43(m,3H),2.55(m,1H),2.90～3.00(m,6H),3.21～3.42(m,7H),3.55(m,1H),3.86(m,2H),4.01～4.26(m,3H),4.66～4.69(m,2H),5.12～5.22(m,2H),5.52(m,1H),6.54(d,1H),7.52(m,2H),7.75(m,1H)。

Example 14: (S) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (3-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-14)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-19) obtained in preparation example 6-19 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-14): 0.18g (60%).

1H NMR(400MHz,CDCl3):0.78～0.99(m,19H),1.29～1.42(m,6H),1.69(m,6H),1.95～2.04(m,2H),2.21～2.43(m,3H),2.55(m,1H),2.90～3.00(m,6H),3.21～3.42(m,7H),3.55(m,1H),3.86(m,2H),4.01～4.26(m,3H),4.66～4.69(m,2H),5.12～5.22(m,2H),5.52(m,1H),6.54(d,1H),7.16(m,1H),7.34(m,5H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-14): 0.12g (94%).

1H NMR(400MHz,CDCl3):0.78～0.99(m,19H),1.29～1.42(m,6H),1.69(m,6H),1.95～2.04(m,2H),2.21～2.43(m,3H),2.55(m,1H),2.90～3.00(m,6H),3.21～3.42(m,7H),3.55(m,1H),3.86(m,2H),4.01～4.26(m,3H),4.66～4.69(m,2H),5.12～5.22(m,2H),5.52(m,1H),6.54(d,2H),7.16(m,2H)。

Example 15: (S) -N- ((3R,4S,5S) -3-methoxy-1- ((2R,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-3- (((R) -1- (4-methoxyphenyl) -1-oxopropan-2-yl) amino) -2-methyl-3-oxopropyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-15)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-20) obtained in preparation example 6-20 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-15): 0.26g (62%).

1H NMR(400MHz,CDCl3):0.73～0.81(m,19H),0.90～0.96(m,6H),1.25～1.30(m,3H),1.34～1.42(m,7H),1.95～2.04(m,2H),2.21～2.43(m,3H),2.55(m,1H),2.90～3.00(m,6H),3.21～3.42(m,7H),3.55(m,1H),3.86(m,2H),4.01～4.26(m,3H),4.66～4.69(m,2H),5.12～5.22(m,2H),5.52(m,1H),6.54(d,1H),6.95～7.05(m,3H),7.30～7.34(m,5H),7.95～7.98(m,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-15): 0.21g (95%).

1H NMR(400MHz,CDCl3):0.73～0.81(m,19H),0.90～0.96(m,6H),1.25～1.30(m,3H),1.34～1.42(m,7H),1.95～2.04(m,2H),2.21～2.43(m,3H),2.55(m,1H),2.90～3.00(m,6H),3.21～3.42(m,7H),3.55(m,1H),3.86(m,2H),4.01～4.26(m,3H),4.66～4.69(m,2H),5.12～5.22(m,2H),5.52(m,1H),6.54(d,2H),7.95～7.98(d,2H)。

Example 16: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((S) -1- (3, 5-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-16).

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-17) obtained in preparation example 6-17 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-16): 0.43g (61%).

1H NMR(400MHz,CDCl3):0.79～1.02(m,20H),1.29(m,3H),1.34～1.44(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,3H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.18(m,1H),7.34(m,5H),7.49(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-16): 0.35g (100%).

1H NMR(400MHz,CDCl3):0.79～1.02(m,20H),1.29(m,3H),1.34～1.44(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,3H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.49(m,2H)。

Example 17: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((S) -1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-17).

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-22) obtained in preparation example 6-22 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-17): 0.16g (67%).

1H NMR(400MHz,CDCl3):0.81-0.98(m,20H),1.29(m,3H),1.34-1.44(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,3H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21-5.23(m,2H),5.36(m,1H),7.03(m,1H),7.18(m,1H),7.35(m,5H),7.49(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-17): 0.11g (92%).

1H NMR(400MHz,CDCl3):0.81～0.98(m,20H),1.29(m,3H),1.34～1.44(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,3H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.18(m,1H),7.49(m,1H)。

Example 18: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (2, 6-difluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-18).

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-21) obtained in preparation example 6-21 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-18): 0.16g (67%).

1H NMR(400MHz,CDCl3):0.81～0.98(m,20H),1.29(m,3H),1.34～1.44(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,3H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.18(m,1H),7.35(m,5H),7.49(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-18): 0.07g (92%).

Example 19: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R, Z) -1- (hydroxyimino) -1- (3-methoxyphenyl) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-19).

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-23) obtained in preparation example 6-23 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-19): 0.28g (62%).

1H NMR(400MHz,CDCl3):0.79-0.98(m,20H),1.01(m,4H),1.19～1.27(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.15(m,2H),7.30(m.1H),7.35(m,5H),7.49(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-19): 0.17g (94%).

1H NMR(400MHz,CDCl3):0.79～0.98(m,20H),1.01(m,4H),1.19～1.27(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,3H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.15(m,2H),7.49(m,1H)。

Example 20: (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (hydroxyimino) -1- (3-methoxyphenyl) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-20).

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in production example 6-1 were not used, but the compound (X-23) and the compound (II-2) obtained in production example 6-23 were used, respectively, to obtain the title compound: 0.23g (58%).

1H NMR(400MHz,CDCl3):0.76～0.98(m,20H),1.01～1.12(m,4H),1.19～1.27(m,4H),2.21～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,6H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.21～5.23(m,2H),5.36(m,1H),7.03(m,1H),7.15(m,2H),7.49(m,1H)。

Example 21: (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- (((R) -1- (4-fluorophenyl) -1- (hydroxyimino) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-21).

The same procedure as in example 1 was repeated except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but (2R,4R) -tert-butyl-2- ((1R,2R) -3- (((R) -1- (4-fluorophenyl) -1- (hydroxyimino) propan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate and the compound (II-2) were used, respectively, to obtain the title compound: 0.17g (63%).

1H NMR(400MHz,CDCl3):0.76～0.98(m,20H),1.01～1.12(m,4H),1.17～1.27(m,4H),2.25～2.43(m,4H),2.77(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,6H),3.61(m,1H),3.99～4.14(m,3H),4.15～4.37(m,3H),4.70(m,1H),5.19～5.23(m,2H),5.36(m,1H),7.20(t,2H),8.01～8.05(m,2H)。

Example 22: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (5-bromo-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-22)

The same procedure as in example 1 was repeated except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (5-bromo-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate and the compound (II-2) were used, respectively, to obtain the title compound: 35mg (36%).

1H NMR(400MHz,CDCl3):0.73～0.97(m,20H),1.01～1.12(m,4H),1.19～1.23(m,4H),2.28～2.42(m,4H),2.52(s,3H),2.77(m,2H),3.03(m,2H),3.30(s,6H),3.34(s,3H),3.42(s,6H),3.59(m,1H),3.01～4.14(m,3H),4.17～4.37(m,3H),4.70(m,1H),5.19～5.23(m,2H),5.36(m,1H),7.15(m,1H),7.65(m,2H)。

Example 23: (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-23)

The same procedure as in example 1 was repeated except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate and the compound (II-2) were used, respectively, to obtain the title compound: 0.14g (74%).

1H NMR(400MHz,CDCl3):0.76～0.95(m,20H),1.01～1.12(m,4H),1.17～1.23(m,4H),2.25～2.42(m,4H),2.48(s,3H),2.77(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,6H),3.59(m,1H),3.01～4.14(m,3H),4.17～4.37(m,3H),4.70(m,1H),5.19～5.23(m,2H),5.36(m,1H),7.20(m,1H),7.71(m,2H)。

Example 24: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-24).

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used but (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of compound (I-24): 0.23g (77%).

1H NMR(400MHz,CDCl3):0.70～0.91(m,14H),0.96～1.12(m,3H),1.32(d,J＝6.8Hz,2H),1.74～2.08(m,4H),2.19～2.60(m,3H),2.68(s,3H),2.89～2.92(m,3H),2.93～3.01(m,3H),3.23(s,3H),3.36(s,3H),3.38～3.46(m,1H),3.49(s,3H),3.87(s,3H),4.01(dd,J＝7.6Hz,2.4Hz,1H),4.03～4.06(m,1H),4.11～4.18(m,2H),4.23-4.28(m,2H),4.60～4.64(m,2H),4.64～4.72(m,1H),5.04～5.25(m,2H),6.64(dd,J＝13.2Hz,2.0Hz,2H),6.78(dd,J＝8.8Hz,2.4Hz,2H),7.34(m,5H),7.87～7.98(m,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-24): 0.18g (91%).

MALDI-TOF MS m/z:803[M+23]+。

1H NMR(400MHz,CDCl3):0.73～0.92(m,20H),1.01～1.12(m,4H),1.17～1.23(m,4H),2.25～2.42(m,4H),2.48(s,3H),2.77(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,3H),3.59(m,1H),3.01～4.14(m,3H),4.17～4.37(m,3H),4.70(m,1H),5.19～5.23(m,2H),5.36(m,1H),7.20(m,1H),7.71(m,2H)。

Example 25: (S) -N- ((3R,4S,5S) -1- ((2S,4R) -2- ((1R,2R) -3- (((E) -2- (2-fluoro-4-methoxyphenyl) -2-hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-25).

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used, but (2S,4R) -tert-butyl-2- ((1R,2R) -3- (((E) -2- (2-fluoro-4-methoxyphenyl) -2- (hydroxyimino) ethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of the compound (I-25): 0.35g (65%).

1H NMR(400MHz,CDCl3):0.72～0.92(m,14H),0.96(d,J＝6.4Hz,2H),1.02(d,J＝6.8Hz,2H),1.32(d,J＝6.8Hz,2H),1.76～2.10(m,4H),2.19～2.60(m,3H),2.89～2.92(m,3H),2.95～3.01(m,3H),3.24(s,3H),3.34(s,3H),3.35～3.44(m,1H),3.46(s,3H),3.87(s,3H),3.98(dd,J＝7.6Hz,2.4Hz,1H),4.00～4.06(m,1H),4.11～4.15(m,2H),4.24～4.28(m,2H),4.62～4.64(m,2H),4.68～4.70(m,,1H),5.09～5.23(m,2H),6.64(dd,J＝13.2Hz,2.0Hz,2H),6.78(dd,J＝8.8Hz,2.4Hz,2H),7.34(m,5H),7.95(t,J＝8.4Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-25) was used: 0.15g (96%).

1H NMR(400MHz,CDCl3):0.80～0.84(m,3H),0.88～1.03(m,17H),1.32(d,J＝7.2Hz,3H),1.35～1.39(m,2H),1.98～2.10(m,4H),2.35(s,3H),2.36～2.51(m,3H),2.74(m,,1H),3.02(m,3H),3.29(s,3H),3.34(s,3H),3.45(s,3H),3.46～3.56(m,1H),3.87(s,3H),3.99(dd,J＝7.6Hz,2.8Hz,1H),4.10～4.15(m,1H),4.25～4.28(m,1H),4.58～4.64(m,2H),4.73～4.78(m,1H),6.67(m,1H),6.78(qd,J＝10.4Hz,2.8Hz,1H),7.95(m,1H)。

Example 26: (S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-26)

The same procedure as in example 1 was repeated except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate and the compound (II-2) were used, respectively, to obtain the title compound: 0.19g (60%).

1H NMR(400MHz,CDCl3):0.76～0.95(m,20H),1.01～1.12(m,4H),1.17～1.23(m,4H),2.25～2.42(m,4H),2.44(s,3H),2.73(m,2H),3.00(m,2H),3.29(s,6H),3.34(s,3H),3.42(s,6H),3.59(m,1H),3.01～4.14(m,3H),4.17～4.37(m,3H),4.70(m,1H),5.17～5.21(m,2H),5.36(m,1H),7.21(m,1H),7.75～7.81(m,2H)。

Example 27: (S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-27)

The same procedures as in example 1 were repeated except for using (2R,4R) -tert-butyl-2- ((1R,2R) -3- ((2- (4-fluoro-2-methylphenyl) -2- (oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidine-1-carboxylate instead of the compound (X-1) obtained in preparation example 6-1 to obtain 0.51g (68%) of the N-terminal amino protected compound in the compound (I-27).

1H NMR(400MHz,CDCl3):0.70～0.91(m,14H),0.96～1.12(m,3H),1.32(d,J＝6.8Hz,2H),1.74～2.08(m,4H),2.19～2.60(m,3H),2.68(s,3H),2.89～2.92(m,3H),2.93～3.01(m,3H),3.23(s,3H),3.36(s,3H),3.38～3.46(m,1H),3.49(s,3H),3.87(s,3H),4.01(dd,J＝7.6Hz,2.4Hz,1H),4.03～4.06(m,1H),4.11～4.18(m,2H),4.23～4.28(m,2H),4.60～4.64(m,2H),4.64～4.72(m,1H),5.04～5.25(m,2H),6.64(dd,J＝13.2Hz,2.0Hz,2H),6.78(dd,J＝8.8Hz,2.4Hz,2H),7.34(m,5H),7.87～7.98(m,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-27) was used: 0.39g (93%).

MALDI-TOF MS m/z:765[M+1]+。

1H NMR(400MHz,CDCl3):0.73～0.92(m,20H),1.01～1.12(m,4H),1.17～1.21(m,4H),2.23～2.41(m,4H),2.45(s,3H),2.77(m,2H),3.02(m,2H),3.29(s,6H),3.34(s,3H),3.41(s,3H),3.59(m,1H),3.01～4.14(m,3H),4.17～4.37(m,3H),4.70(m,1H),5.19～5.23(m,2H),5.36(m,1H),7.25(m,1H),7.73～7.89(m,2H)。

Example 28: (S) -N- ((3R,4S,5S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2- (o-tolyl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-28)

The same procedure as in example 1 was repeated except that the compound (X-1) obtained in preparation example 6-1 was not used but (2S,4R) -tert-butyl-4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2- (o-tolyl) ethyl) amino) propyl) pyrrolidine-1-carboxylate was used to obtain a compound protected at the N-terminal amino group of compound (I-28): 0.15g (24%).

1H NMR(400MHz,CDCl3):0.76～0.86(m,11H),0.87(d,J＝6.8Hz,6H),0.96(d,J＝6.8Hz,3H),1.33(d,J＝4.8Hz,2H),1.72～2.01(m,4H),2.22～2.60(m,3H),2.43(s,3H),2.89～2.93(m,3H),2.95～3.01(m,3H),3.27(s,3H),3.33(s,3H),3.35～3.41(m,1H),3.46(s,3H),4.01～4.17(m,3H),4.23～4.25(m,2H),4.67～4.69(m,2H),4.72～4.74(m,2H),5.12～5.22(m,2H),7.27(d,J＝8.4Hz,2H),7.29～7.34(m,5H),7.85～7.94(m,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-28): 60mg (46%).

MALDI-TOF MS m/z:747[M+1]+。

1H NMR(400MHz,CDCl3):0.81～0.85(m,3H),0.92～1.03(m,17H),1.26～1.29(m,3H),1.31～1.44(m,2H),1.96～2.08(m,4H),2.35(s,6H),2.38～2.53(m,3H),2.77(d,J＝4.8Hz,,1H),3.03(m,3H),3.29(s,3H),3.33(s,3H),3.34～3.40(m,1H),3.44(d,J＝8.4Hz,3H),3.45～3.55(m,1H),3.80～3.84(m,1H),3.89～3.98(m,1H),4.07(m,2H),4.75～4.79(m,2H),4.85～4.91(m,1H),7.09～7.16(m,2H),7.31～7.38(m,2H)。

Example 29: (S) -N- ((3R,4S,5S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2- (p-tolyl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-29)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-9) obtained in preparation example 6-9 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-29): 0.24g (95%).

1H NMR(400MHz,CDCl3):0.76-0.86(m,11H),0.87(d,J＝6.8Hz,6H),0.96(d,J＝6.8Hz,3H),1.33(d,J＝4.8Hz,2H),1.74～2.03(m,4H),2.20～2.61(m,3H),2.43(s,3H),2.89～2.93(m,3H),2.95～3.01(m,3H),3.27(s,3H),3.33(s,3H),3.35～3.44(m,1H),3.46(s,3H),4.00(dd,J＝7.5Hz,2.8Hz,1H),4.12～4.15(m,2H),4.23～4.25(m,2H),4.67～4.69(m,2H),4.72～4.74(m,2H),5.12～5.22(m,2H),7.27(d,J＝8.4Hz,2H),7.29～7.34(m,5H),7.87(d,J＝8.4Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-29) was used: 0.19g (95%).

1H NMR(400MHz,CDCl3):0.81～0.85(m,3H),0.92～1.03(m,17H),1.26～1.29(m,3H),1.31～1.44(m,2H),1.96～2.10(m,4H),2.33(s,6H),2.37～2.51(m,3H),2.74(d,J＝4.8Hz,1H),3.03(m,3H),3.29(s,3H),3.33(s,3H),3.34～3.40(m,1H),3.42(d,J＝8.4Hz,3H),3.47～3.56(m,1H),3.80～3.84(m,1H),3.91～3.98(m,1H),4.07(m,2H),4.75～4.79(m,2H),4.85～4.91(m,1H),7.15(m,2H),7.30(m,2H)。

Example 30: (S) -N- ((3R,4S,5S) -1- ((2S,4R) -2- ((1R,2R) -3- (2- (4-fluorophenyl) -2-oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-30)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-10) obtained in preparation example 6-10 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-30): 0.25g (97%).

1H NMR(400MHz,CDCl3):0.75～0.86(m,11H),0.88(d,J＝6.4Hz,6H),0.96(d,J＝6.8Hz,3H),1.32(d,J＝7.2Hz,2H),1.76～2.03(m,4H),2.20～2.61(m,3H),2.89～2.93(m,3H),2.95～3.01(m,3H),3.23(s,3H),3.34(s,3H),3.35～3.44(m,1H),3.46(s,3H),4.00(dd,J＝7.5Hz,2.8Hz,1H),4.00～4.06(m,1H),4.12～4.15(m,2H),4.24～4.26(m,2H),4.67～4.69(m,2H),4.72(d,J＝4.8Hz,2H),5.09～5.23(m,2H),7.17(t,J＝8.8Hz,2H),7.34(m,5H),8.00(dd,J＝8.8Hz,5.2Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-30) was used: 0.2g (95%).

1H NMR(400MHz,CDCl3):0.74～0.83(m,4H),0.86～1.03(m,16H),1.25～1.29(m,2H),1.33(d,J＝6.8,3H),1.98～2.10(m,4H),2.33(s,3H),2.37～2.51(m,3H),2.74(d,J＝4.8Hz,1H),3.02(m,3H),3.29(s,3H),3.34(d,J＝6.4Hz,3H),3.35～3.40(m,1H),3.46(s,3H),3.91～4.05(m,2H),4.10～4.17(m,1H),4.18～4.26(m,1H),4.72(d,J＝4.8Hz,2H),4.75～4.82(m,2H),7.10(t,J＝8.8Hz,1H),7.17(t,J＝8.8Hz,1H),7.38(m,1H),8.02(m,1H)。

Example 31: (S) -N- ((3R,4S,5S) -1- ((2S,4R) -2- ((1R,2R) -3- (2- (2-fluoro-4-methoxyphenyl) -2-oxoethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-31)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-7) obtained in preparation example 6-7 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-31): 0.24g (95%).

1H NMR(400MHz,CDCl3)0.72～0.92(m,14H),0.96(d,J＝6.4Hz,2H),1.02(d,J＝6.8Hz,2H),1.32(d,J＝6.8Hz,2H),1.76～2.10(m,4H),2.19～2.60(m,3H),2.89～2.92(m,3H),2.95～3.01(m,3H),3.24(s,3H),3.34(s,3H),3.35～3.44(m,1H),3.46(s,3H),3.87(s,3H),3.98(dd,J＝7.6Hz,2.4Hz,1H),4.00～4.06(m,1H),4.11～4.15(m,2H),4.24～4.28(m,2H),4.62～4.64(m,2H),4.68～4.70(m,,1H),5.09～5.23(m,2H),6.64(dd,J＝13.2Hz,2.0Hz,2H),6.78(dd,J＝8.8Hz,2.4Hz,2H),7.34(m,5H),7.95(t,J＝8.4Hz,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-31): 0.18g (89%).

1H NMR(400MHz,CDCl3):0.80～0.84(m,3H),0.88～1.03(m,17H),1.32(d,J＝7.2Hz,3H),1.35～1.39(m,2H),1.98～2.10(m,4H),2.35(s,3H),2.36～2.51(m,3H),2.74(m,1H),3.02(m,3H),3.29(s,3H),3.34(s,3H),3.45(s,3H),3.46～3.56(m,1H),3.87(s,3H),3.99(dd,J＝7.6Hz,2.8Hz,1H),4.10～4.15(m,1H),4.25～4.28(m,1H),4.58～4.64(m,2H),4.73～4.78(m,1H),6.67(m,1H),6.78(qd,J＝10.4Hz,2.8Hz,1H),7.95(m,1H)。

Example 32: (S) -N- ((3R,4S,5S) -1- ((S) -4- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) thiazolidin-3-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyramido) butanamide (I-32)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-24) obtained in preparation example 6-24 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-32): 0.23g (70%).

1H NMR(400MHz,CDCl3):0.74～1.05(m,18H),1.17～1.25(d,3H),1.47(s,9H),1.61(s,2H),2.21～2.47(m,3H),2.65～3.12(m,8H),2.88(s,3H),2.98(s,3H),3.35(s,3H),3.42(S,3H),3.77～4.18(m,3H),4.16～4.21(m,3H),4.66～4.70(m,2H),5.09～5.23(m,3H),7.32(m,7H),7.59(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound (I-32) whose N-terminal amino group was protected was used: 0.17g (90%).

MALDI-TOF MS m/z:742.9[M+1]+。

1H NMR(400MHz,CDCl3):0.74～1.05(m,18H),1.17～1.25(d,3H),1.47(s,9H),1.61(s,2H),2.21～2.47(m,3H),2.65～3.12(m,8H),2.88(s,3H),2.98(s,3H),3.35(s,3H),3.42(S,3H),3.77～4.18(m,3H),4.16～4.21(m,3H),4.66～4.70(m,2H),5.16(br,1H),7.32～7.37(m,2H),7.59(m,1H)。

Example 33: (2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4S) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- (phenethylamino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-33)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-11) obtained in preparation example 6-11 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-33): 0.40g (77%).

1H NMR(400MHz,CDCl3):0.79～1.06(m,15H),1.73～1.38(m,5H),1.76～2.03(m,6H),2.98～2.37(m,4H),2.77～2.82(m,2H),2.90～3.01(m,3H),3.0(s,3H),3.25～3.28(m,1H),3.3(s,3H),3.33(s,3H),3.36(s,3H),3.38～3.54(m,3H),3.74～3.75(m,1H),3.81～3.96(m,1H),3.93～3.96(m,1H),4.09～4.11(m,1H),4.68～4.69(m,1H),5.09～5.23(m,2H),6.16～6.17(m,1H)6.53～6.54(m,1H),7.17～7.34(m,10H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-33): 0.31g (93%).

1H NMR(400MHz,CDCl3):0.81～0.85(m,3H),0.92～1.16(m,15H),1.16～1.24(d,3H),1.27～1.37(m,1H),1.89～2.12(m,5H),2.33(s,3H),2.36～2.37(m,3H),2.74～2.96(m,4H),3.03(s,3H),3.23～3.28(m,1H),3.31(s,3H),3.36(s,3H),3.4(s,3H),3.51～3.62(m,3H),3.7～3.78(m,1H),3.81～3.96(m,1H),3.91～3.97(m,1H),4.11～4.15(m,2H)4.44～4.88(m,2H),6.2(m,1H),7.17～7.34(m,5H),7.57～7.60(m,1H)。

EXAMPLE 34 (2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2-oxo-2-phenylethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-34)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-8) obtained in preparation example 6-8 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-34): 2.36g (81%).

1H NMR(400MHz,CDCl3):0.71～1.04(m,17H),1.31～1.32(m,4H),1.77～1.81(m,2H)1.92～2.04(m,4H),2.17～2.61(m,4H),2.89～2.95(m,3H),2.99(s,3H),3.31(s,6H),3.46(s,3H),3.43-3.52(m,1H),3.99～4.01(m,1H),4.09～4.15(m,1H),4.24(m,1H),4.69～4.73(m,4H),4.74～4.76(d,2H),5.09～5.23(m,2H),6.46～6.49(m,1H)7.12(m,1H),7.26～7.34(m,5H),7.48～7.51(t,2H),7.6～7.64(t,1H),7.96～8.04(d,2H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-34): 1.89g (96%).

1H NMR(400MHz,CDCl3):0.81～0.85(m,3H),0.92～1.14(m,13H),1.25～1.43(m,5H),1.46～1.89(m,6H),1.92～2.22(m,5H),2.33(s,3H),2.45～2.54(m,2H),2.73～2.74(d,1H),3.02(s,3H),3.32(s,6H),3.41(d,3H),3.52～3.84(m,3H),3.92～4.06(m,3H),4.75～4.8(m,2H),4.94～5.23(m,1H),6.76～6.94(m,1H),7.34～7.43(m,4H),7.60(m,1H)。

Example 35: (2S) -N- ((3R,4S) -1- ((2S,4R) -2- ((1R,2R) -3- (2- (2-hydroxy-2-phenylethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-35)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-12) obtained in preparation example 6-12 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-35): 0.24g (57%).

1H NMR(400MHz,CDCl3):0.71～0.97(m,17H),1.22～1.29(m,4H),1.77～1.81(m,2H)1.92～2.04(m,4H),2.17～2.61(m,4H),2.88～2.95(m,3H),2.99(s,3H),3.32(s,6H),3.46(s,3H),3.43～3.52(m,1H),3.52～4.04(m,5H),4.51～4.96(m,5H),5.09～5.23(m,2H),6.46～6.49(m,1H)7.12(m,1H),7.26～7.43(m,10H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-35): 0.20g (99%).

1H NMR(400MHz,CDCl3):0.81～0.86(m,3H),0.93～1.03(m,12H),1.18(d,1H),1.20～1.31(m,3H),1.53～1.79(m,6H),2.01～2.08(m,3H),2.35(s,3H),2.34～2.39(m,1H),2.73～2.74(d,1H),3.03(d,1H),3.06(s,2H),3.32(s,6H),3.38～3.48(m,3H),3.92～4.18(m,5H),4.75～4.95(m,2H),7.23～7.4(m,5H)。

Example 36: (2S) -N- ((3R,4S) -1- ((2S) -2- ((1R,2R) -3- (((2R) -1-amino-1-phenylpropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-36)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 and the compound (II-1) were not used, but the compound (X-13) obtained in preparation example 6-13 and the compound (II-2) were used, respectively, to obtain a compound in which the N-terminal amino group of the compound (I-36) was protected: 55mg (52%).

1H NMR(400MHz,CDCl3):0.79～0.81(m,3H)0.92～1.09(m,12H),1.11～1.25(m,6H)1.72～2.24(m,9H),2.24(s,3H),2.45(m,2H),2.98-2.93(m,3H),3.33(s,6H),3.35(s,3H),3.45～3.54(m,2H),3.71(d,1H),4.01-4.15(m,2H),4.23-4.37(m,2H),4.69～4.75(m,2H),5.12-5.22(m,2H),6.74～6.88(m,1H),7.26～7.33(m,10H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-36): 44mg (95%).

MALDI-TOF MS m/z:731.6[M+1]+

Example 37: (2S) -N- ((3R,4S) -1- ((2S,4R) -2- ((1R,2R) -3- (((2S) -1-amino-1-phenylpropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-37)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 and the compound (II-1) were not used, but the compound (X-14) obtained in preparation example 6-14 and the compound (II-2) were used, respectively, to obtain a compound in which the N-terminal amino group of the compound (I-37) was protected: 96mg (48%).

1H NMR(400MHz,CDCl3):0.79～0.81(m,3H),0.92～1.09(m,12H),1.11～1.25(m,6H),1.72～2.24(m,9H),2.24(s,3H),2.45(m,2H),2.98-2.93(m,3H),3.33(s,6H),3.35(s,3H),3.45～3.54(m,2H),3.71(d,1H),4.01-4.15(m,2H),4.23-4.37(m,2H),4.69～4.75(m,2H),5.12-5.22(m,2H),6.74～6.88(m,1H),7.26～7.33(m,10H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-37) was used: 75mg (95%).

MALDI-TOF MS m/z:730.9[M+1]+。

Example 38: (2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2- (pyridin-2-yl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-38)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-15) obtained in preparation example 6-15 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-38): 0.35g (35%).

1H NMR(400MHz,CDCl3):0.75～1.00(m,18H),1.19～1.24(m,3H),1.94～1.97(m,4H),2.04(s,1.H),2.23～2.26(m,2H),2.35～2.49(m,2H),2.87～2.92(m,3H),2.96～3.12(m,3H),3.31(s,4H),3.37(d,3H),3.42～3.49(m,3H),3.61～3.68(m,3H),3.86～3.89(m,1H),4.11～4.19(m 1H),4.70(m,1H),5.10～5.23(m,2H),6.92(m,1H),7.12～7.20(m,2H),7.33(m,5H),7.58～7.63(m,2H),8.51～8.21(m,1H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-38): 0.25g (86%).

1H NMR(400MHz,CDCl3):0.81～0.84(m,3H),0.92～1.00(m,15H),1.19～1.25(m,3H),1.76～2.03(m,4H),2.34(m,3H),2.37～2.42(m,1H),2.27～2.77(m,1H),2.99～3.03(m,3H),3.15(s,1H),3.32(s,6H),3.38(d,J＝4Hz,3H),3.64(m,2H),3.88(m,1H),4.12(m,2H),4.77(m,1H),6.93(m,1H),7.12～7.19(m,2H),7.58～7.65(m,2H),8.52(m,1H)。

Example 39: (2S) -N- ((3R,4S) -3-methoxy-1- ((2S,4R) -4-methoxy-2- ((1R,2R) -1-methoxy-2-methyl-3-oxo-3- ((2- (thien-2-yl) ethyl) amino) propyl) pyrrolidin-1-yl) -5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-39)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-16) obtained in preparation example 6-16 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-39): 0.8g (99%).

1HNMR(400MHz,CDCl3):0.79～1.00(m,18H),1.01～1.25(m,3H),1.75～1.82(m,4H),2.04(s,1.H),2.24～2.42(m,2H),2.35～2.49(m,2H),2.88～2.92(m,3H),3.00～3.07(m,4H),3.27(s,3H),3.33(m,3H),3.38(m,3H),3.45～3.57(m,3H),3.85～3.87(m,1H),4.08～4.16(m,1H),4.71(m,1H),5.13～5.23(m,2H),6.57(m,1H),6.84(d,J＝2.8Hz,1H),6.91～6.93(m,1H),7.13(d,J＝2.1Hz,1H),7.34(m,5H)。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-39): 70mg (10%).

1H NMR(400MHz,CDCl3):0.81～0.85(m,3H),0.88～1.00(m,15H),1.22～1.26(m,3H),2.03(m,3H),2.34～2.37(m,3H),3.01～3.07(m,3H),3.15(s,4H),3.33(d,3H),3.38(d,3H),3.43～3.58(m,2H),3.85～3.87(m,1H),4.09～4.13(m,2H),4.75～4.79(m,1H),6.62(m,1H),6.90(m,1H),6.91～6.93(m,1H),7.13～7.20(m,1H)。

Example 40: (2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-40)

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but the compound (X-25) and (3R,4S,5S) -4- ((S) -2- (dimethylamino) -3-methylbutanamido) -N, 3-dimethylbutyrylamino) -3-methoxy-5-methylheptadecanoic acid (compound II-2) obtained in preparation example 6-25 were used, respectively, to obtain the title compound: 2.25g (76%).

[α]D 25＝-40.5°(c＝1,MeOH)。

1H NMR(400MHz,CDCl3):0.79-1.05(m,19H),1.26(m,3H),1.45(m,2H),1.66(m,2H),1.97-2.17(m,3H),2.44-2.46(m,3H),2.71-2.80(m,3H),2.81-2.89(m,1H),2.91-2.94(m,2H),2.71-2.80(m,3H),3.04(s,3H),3.26-3.33(m,2H),3.37(s,3H),3.74(m,2H),4.28-4.31(m,2H),4.75-4.82(m,2H),6.18(m,1H),6.84-6.89(m,2H),7.15-7.19(m,1H),7.58(d,1H)。

LC-MS m/z:754[M+]+。

Example 41: (2S) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethylbutanamide (I-41)

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but the compound (X-26) and the (3R,4S,5S) -4- ((S) -2- (dimethylamino) -3-methylbutanamino) -N, 3-dimethylbutyrylamino) -3-methoxy-5-methylheptanoic acid (compound II-2) obtained in preparation example 6-26 were used, respectively, to obtain the title compound: 3.4g (58%).

1H NMR(400MHz,CDCl3):0.80～0.83(m,3H),0.92～1.04(m,9H),1.22～1.31(m,3H),1.35～1.42(m,1H),1.43～1.45(m,3H),1.97～2.09(m,3H),2.21～2.22(m,1H),2.30～2.38(m,6H),2.20～2.58(m,3H),2.72～2.76(m,1H),3.01(d,2H),3.35(d,3H),3.35～3.62(m,1H),3.55(d,3H),4.08～4.15(m,2H),4.31～4.37(m,1H),4.73～4.79(m,1H),5.43～5.54(m,1H),7.15～7.17(m,2H),7.53～7.61(m,1H),8.02～8.09(m,2H)。

LC-MS m/z:764[M+]+。

Example 42: (2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butanamide (I-42)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-25) obtained in preparation example 6-25 was used, to obtain a compound protected at the N-terminal amino group of the compound (I-42): 4.37g (63%).

ES-MS m/z:874[M+H]+。

The title compound was prepared by the same manner as in example 1 except that the compound whose N-terminal amino group was protected of the compound (I-42): 2.98g (85%).

1H NMR(400MHz,CDCl3):0.80～0.85(m,3H),0.92～1.00(m,15H),1.01～1.08(m,1H),1.26(d,3H,J＝7.2Hz),1.31～1.38(m,1H),1.81～2.17(m,4H),2.36(s,3H),2.40～2.42(m,2H),2.76～2.78(m,1H),2.90～2.93(m,2H),3.03(s,3H),3.31(s,3H),3.35～3.57(m,4H),3.49(s,3H),4.03～4.10(m,2H),4.28～4.30(m,2H),4.75～4.89(m,2H),6.21(m,1H),6.82～6.91(m,2H),7.13～7.21(m,1H),7.58(d,1H,J＝9.2Hz)。

LC-MS m/z:740[M+]+。

Example 43: (2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- (((S) -1- (4-fluorophenyl) -1-oxopropan-2-yl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-hydroxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-43)

The same procedure as in example 1 was repeated, except that the compound (X-1) obtained in preparation example 6-1 was not used, but the compound (X-26) obtained in preparation example 6-26 was used, to obtain the title compound: 0.12g (51%).

1H NMR(400MHz,CDCl3):0.77～0.83(m,3H),0.92～1.04(m,9H),1.32～1.35(m,3H),1.34～1.41(m,1H),1.41～1.42(m,3H),1.97～2.09(m,3H),2.21～2.22(m,1H),2.33～2.35(m,3H),2.23～2.56(m,3H),2.72～2.76(m,1H),3.01(d,2H),3.35(d,3H),3.35～3.62(m,1H),3.55(d,3H),4.08～4.15(m,2H),4.31～4.37(m,1H),4.73～4.79(m,1H),5.43～5.54(m,1H),7.15～7.17(m,2H),7.57～7.60(m,1H),8.00～8.07(m,2H)。

LC-MS m/z:750[M+]+,772[M+Na]+。

EXAMPLE 44 (2S) -N- ((3R,4S) -1- ((2S,4S) -4-amino-2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) pyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-44)

The same procedure as in example 1 was repeated, except that the compound (X-1) and the compound (II-1) obtained in preparation example 6-1 were not used, but the compound (X-27) and the (3R,4S,5S) -4- ((S) -2- (dimethylamino) -3-methylbutanamido) -N, 3-dimethylbutyrylamino) -3-methoxy-5-methylheptanoic acid (compound II-2) obtained in preparation example 6-27 were used, respectively, to obtain an azide compound of the compound (I-44): 0.42g (76%).

1H NMR(400MHz,CDCl3):0.79-1.07(m,15H),1.20(d,2H),1.99-2.11(m,4H),2.18-2.28(m,5H),2.33-2.45(m,4H),2.90-3.27(m,6H),3.32-3.42(m,6H),3.47-3.54(m,2H),3.75-3.78(m,1H),3.85-3.98(m,2H),4.19-4.21(m,1H),4.39-4.44(m,1H),4.46-4.80(m,1H),4.87-4.90(m,1H),6.17(m,1H),6.84-6.95(m,2H),7.14-7.26(m,1H)。

The azide compound of the compound (I-44) was dissolved in 10mL of methanol, and the reaction was carried out in the presence of 10% palladium on carbon (15mg) under a hydrogen atmosphere while stirring. After completion of the reaction, the reaction mixture was filtered through celite, and washed several times with methanol. The solvent was removed under vacuum to leave the title compound as a white solid: 0.23g (99%).

1H NMR(400MHz,CDCl3):0.79-1.03(m,14H),1.21-1.22(m,5H),2.06-2.45(m,6H),2.89-3.02(m,4H),3.30-3.44(m,5H),3.47-3.52(m,2H),3.67-3.69(m,1H),3.71-4.39(m,3H),4.78-4.80(m,1H),6.17(m,1H),6.84-6.95(m,2H),7.14-7.26(m,1H)。

LC-MS m/z:753[M+]+,775[M+Na]+。

Example 45: (2S) -N- ((3R,4S) -1- ((2S,4S) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4- (methylamino) pyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -2- ((S) -2- (dimethylamino) -3-methylbutanamide) -N, 3-dimethylbutanamide (I-45)

The same procedure as in example 1 was repeated, except that the compound (X-28) obtained in production example 6-28 and (3R,4S,5S) -4- ((S) -2- ((S) -2- (dimethylamino) -3-methylbutanamido) -N, 3-dimethylbutanoylamino) -3-methoxy-5-methylheptanoic acid (compound II-2) obtained in production example 6-28 were used instead of the compound (X-1) and the compound (II-1) obtained in production example 6-1, respectively, to obtain an amino-protected compound of compound (I-45): 0.13g (64%).

LC-MS m/z:857.5[M+]+。

The amino-protected compound of the compound (I-45) was dissolved in 10mL of methanol and reacted in the presence of 20% palladium hydroxide (66mg) under hydrogen pressure (55psi) for 12 hours while stirring. After completion of the reaction, the reaction mixture was filtered through celite, and washed several times with methanol. The solvent was removed under vacuum to leave the title compound as a white solid: 115mg (100%).

LC-MS m/z:767.6[M+]+,789.4[M+Na]+。

Test example 1: determination of anticancer Activity

Compounds of the invention were assayed for inhibitory activity in 7 cancer cell lines, which were: breast cancer (BT-474), human epithelial cancer (A-431), non-small cell lung cancer (NCI-H460), colon cancer (HT-29), colon cancer (SW-620), breast cancer (MDA-MB-231), and breast cancer (MCF-7).

Briefly, suspensions of each cell line were serially diluted and treated at 2-3 × 10³Cell/well Density seeded on 96-well microplates followed by 37 ℃ and 5% CO₂Incubate under atmosphere for 4 days. Cell growth in the monolayer was measured using WST-8(Dojindo, Japan) reagent.

IC50 values of compounds against cancer cell lines were calculated from the concentration of compounds grown by cells under the control of 50% OD. The results are summarized in table 1 below.

Meanwhile, the present invention is not limited to the above-described embodiments, and modifications and variations may be made without departing from the gist of the present invention, and it is to be understood that technical inspiration of these modifications and variations also falls within the scope of the appended claims.

TABLE 1

Claims

1. A dolastatin-10 derivative represented by the following chemical formula I, or a pharmaceutically acceptable salt thereof,

[ chemical formula I ]

Wherein,

R₁、R₂、R₃、R₄and R₅Each independently is hydrogen or C₁-C₄Alkyl radical；

ar is unsubstituted thienyl, pyridyl, phenyl, or thienyl, pyridyl, phenyl substituted with at least one member selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, and halo;

x is a carbon atom, an oxygen atom or a sulfur atom; and

2. Dolastatin-10 derivatives or pharmaceutically acceptable salts thereof according to claim 1,

R₁、R₃and R₄Each is C₁-C₄An alkyl group;

R₂and R₅Each independently is hydrogen or C₁-C₄An alkyl group;

ar is unsubstituted phenyl, or is selected from C₁-C₄Alkyl radical, C₁-C₄Phenyl substituted with at least one of the group consisting of alkoxy and halogen;

x is a carbon atom; and

R₇is hydroxy, amino, C₁-C₄Alkoxy or C₁-C₄An alkylamino group.

3. Dolastatin-10 derivatives or pharmaceutically acceptable salts thereof according to claim 1,

R₁、R₃and R₄Each is methyl;

R₂and R₅Each independentlyIs hydrogen or methyl;

R₆is hydrogen, hydroxy, methoxy, amino, oxo (═ O) or oximino (═ N-OH);

x is a carbon atom; and

R₇is hydroxyl, amino, methoxy or N-methylamino.

4. A dolastatin-10 derivative represented by the following chemical formula Ia, or a pharmaceutically acceptable salt thereof,

[ chemical formula Ia ]

Wherein,

x is a carbon atom, an oxygen atom or a sulfur atom; and

5. Dolastatin-10 derivatives or pharmaceutically acceptable salts thereof according to claim 1, selected from the group consisting of:

(S) -N- ((3R,4S,5S) -1- ((2R,4R) -2- ((1R,2R) -3- ((2, 6-difluorophenethyl) amino) -1-methoxy-2-methyl-3-oxopropyl) -4-methoxypyrrolidin-1-yl) -3-methoxy-5-methyl-1-oxoheptan-4-yl) -N, 3-dimethyl-2- ((S) -3-methyl-2- (methylamino) butyrylamino) butyramide (I-1);

6. A method for producing dolastatin 10 derivatives represented by the following chemical formula I, which comprises subjecting a compound represented by the following chemical formula II and a compound represented by the following chemical formula III to a condensation reaction, and deprotecting the condensation product when a protecting group is present,

[ chemical formula II ]

[ chemical formula III ]

[ chemical formula I ]

Wherein,

R₃、R₄and R₅Each independently is hydrogen or C₁-C₄An alkyl group;

x is a carbon atom, an oxygen atom or a sulfur atom; and

7. The method of claim 6, wherein the amino protecting group is t-butyloxycarbonyl (t-Boc), benzyloxycarbonyl (Cbz), fluorenylmethyloxycarbonyl (Fmoc), or benzyl (Bn).

8. The process according to claim 6, wherein the protected hydroxyl group is protected by a protecting group, tert-butyldimethylsilyl.

9. The method of claim 6, wherein the compound of formula III is in the form of a hydrochloride salt or a trifluoroacetate salt (TFA).

10. The process according to claim 6, wherein the condensation reaction is carried out in the presence of a coupling agent selected from the group consisting of Dicyclohexylcarbodiimide (DCC), azidodiphenyl phosphate (DPPA), diethyl cyanophosphate (DEPC) and benzotriazol-1-yl-oxy-tris (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) and an organic base selected from the group consisting of triethylamine and Diisopropylethylamine (DIPEA).

11. The method of claim 6, wherein the amino protecting group is deprotected using 10% palladium on carbon or 20% palladium hydroxide.

12. The method of claim 6, wherein deprotection of the hydroxyl protecting group is performed using tetrabutylammonium fluoride.

13. An anticancer pharmaceutical composition comprising dolastatin-10 derivative according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

14. The anticancer pharmaceutical composition of claim 13, wherein the anticancer drug is used to treat breast cancer.