KR20190101937A - Amatoxin derivatives and methods for their preparation - Google Patents

Abstract

In one aspect of the present invention, provided are amatoxin derivatives, methods for preparing the same, synthetic intermediates of amatoxin derivatives, and a pharmaceutical composition for the treatment of cancer including amatoxin derivatives. Provided in the present invention are a compound represented by formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

Description

Amatoxin derivatives and methods for preparing the same

The present invention relates to a matotoxin derivative, a method for preparing the same, a synthetic intermediate of the matotoxin derivative, and a pharmaceutical composition for treating cancer, including amatetoxin derivative.

Amatoxins are cyclic peptides consisting of eight amino acids. These can be isolated from, for example, Amanita phalloides mushrooms or can be prepared synthetically. Amatoxins specifically inhibit DNA-dependent RNA polymerase II of mammalian cells, thereby inhibiting transcription and protein activity of infected cells. Inhibiting transcription in cells stops cell growth and proliferation.

Although not covalently bound, the complex between anithine and RNA polymerase II is very tight (K _D = 3 nM). Dissociation of anithin from the enzyme is a very slow process and therefore, it is difficult for the infected cells to recover. If inhibition of transcription persists too long, the cells will progress to predetermined cell death (apoptosis).

The use of amatoxins as cytotoxic powders for tumor treatment was developed in 1981 by coupling anti-Thy 1,2 antibodies to α-amanitin using a linker attached to the indole ring of Trp via diazotization. It has been. Davis and Preston identified the attachment site in the 7 'position. In addition, Morris and Venton have published derivatives in which cytotoxic activity is maintained by substitution at the 7 'position.

Meanwhile, the anti-cancer drug industry is currently attracting not only the 'immune anti-cancer drug' and 'anti-immune target anti-cancer drug', but also the ADC (Antibody-Drug Conjugate) technology as a new technology of antibody treatment. ADC technology consists of drugs, monoclonal antibodies, and linkers linking antibodies and drugs, highlighting the merits of antibodies and drugs, and making it possible to target only specific cells. (Non-Patent Document 1).

In this case, in order to bind the drug to the monoclonal antibody, not only an appropriate linker, but also a suitable functional group on the surface of the drug exhibiting a therapeutic effect is preferable.

Thus, the inventors of the present inventors while research to utilize amatoxin in the ADC technology,

The amatoxin derivative provided in one aspect of the present invention may be usefully used as an ADC drug due to the presence of an amine group in the indole group. Experiments have demonstrated that it can be prepared via intermediates of very different structures and have completed the present invention.

Nature Reviews Drug Discovery 16, 315-337 (2017)

An object in one aspect of the present invention is to provide an amatoxin derivative.

Another object of the present invention is to provide a method for preparing the amatoxin derivative.

It is an object in another aspect of the present invention to provide an intermediate synthesized in the preparation of the amatoxin derivative.

Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of cancer containing the amatoxin derivative as an active ingredient.

In order to achieve the above object,

In one aspect of the invention there is provided a compound represented by formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula I]

(In Formula I,

n is an integer from 0 to 2;

R ¹ is —H, —OH, —OR, —SH, —SR, —NH ₂ , —NHR, or —NR ₂ ,

Wherein R is C _1-10 straight or branched chain alkyl, or a protecting group; And

R ² , R ³ , R ⁴ and R ⁵ are independently —H, —OH, —NH ₂ , or C _1-10 straight or branched chain alkyl).

In another aspect of the invention, as shown in Scheme 1 below,

Reacting the compound represented by the formula (II) with the compound represented by the formula (III) to prepare a compound represented by the formula (IV);

Preparing a compound represented by Formula V from the compound represented by Formula IV prepared in the above step;

Preparing a compound represented by Chemical Formula VI through a cyclization reaction through dehydration of the compound represented by Chemical Formula V prepared in the above step; And

There is provided a process for preparing a compound represented by the formula (I), which comprises the step of preparing a compound represented by the formula (I) by removing a protecting group from the compound represented by the formula (VI) prepared in the step.

Scheme 1

(In Scheme 1,

L is absent, -O-, -NH-, -NR-, or -S-;

n, R, R ¹ to R ⁵ are independently as defined in formula (I) above; And

PG ₁ to PG ₃ are protecting groups.

In another aspect of the present invention, there is provided a compound represented by the following formula α, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula α]

(In the above formula α,

n and L are as defined in Scheme 1 above; And

A ¹ and A ² are independently —H, or a protecting group).

In another aspect of the present invention, there is provided a compound represented by the following formula β, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula β]

(In the above formula β,

R ³ is as defined in Formula I above; And

E ¹ , E ² and E ³ are independently —H, —OH, or a protecting group).

In another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating cancer containing the compound represented by the formula (I), the optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The amatoxin derivative provided in one aspect of the present invention may be usefully used as an ADC drug due to the presence of an amine group in the indole group. Can be prepared via intermediates of very different structures.

Hereinafter, the present invention will be described in detail.

In one aspect of the present invention, there is provided a compound represented by the following formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula I]

(In Formula I,

n is an integer from 0 to 2;

R ¹ is —H, —OH, —OR, —SH, —SR, —NH ₂ , —NHR, or —NR ₂ ,

Wherein R is C _1-10 straight or branched chain alkyl, or a protecting group; And

R ² , R ³ , R ⁴ and R ⁵ are independently —H, —OH, —NH ₂ , or C _1-10 straight or branched chain alkyl).

On the other side,

n is an integer from 0 to 2;

R ¹ is —H, —OH, —OR, —SH, —SR, —NH ₂ , —NHR, or —NR ₂ ,

Wherein R is C _1-5 straight or branched alkyl, or a protecting group; And

R ² , R ³ , R ⁴ and R ⁵ are independently —H, —OH, —NH ₂ , or C _1-5 straight or branched chain alkyl.

In another aspect,

n is an integer from 0 to 2;

R ¹ is —H, —OH, —OR, —SH, —SR, —NH ₂ , —NHR, or —NR ₂ ,

Wherein R is C _1-3 straight or branched alkyl, or a protecting group;

R ² is -NH ₂ , or -OH; And

R ³ , R ⁴ and R ⁵ are independently -H, or -OH.

In another aspect, the compound represented by Formula I may be a compound having the following structure.

.

.

Meanwhile, the protecting group may be used without limitation as long as it is a protecting group known in the compound synthesis field, and for example, acetyl (Acetyl, Ac), acetoxy (Acetoxy, OAc), β-methoxyethoxy Β-Methoxyethoxymethyl ether (MEM), Dimethoxytrityl, Dimethoxytrityl, Methoxymethyl ether (MOM), Methoxytrityl, p-Methoxybenzyl ether, PMB), methylthiomethyl ether, pivaloyl (Piv), triphenylmethyl, tert-Butyloxycarbonyl (Boc), 9-fluorenylmethyloxycarbon Nyl (9-Fluorenylmethyloxycarbonyl, Fmoc), Tosyl, Ts, tert-Butyl esters.

In another aspect of the present invention, the compound represented by the formula (I) may be used in the form of a pharmaceutically acceptable salt, the salt is an acid addition salt formed by a pharmaceutically acceptable free acid is useful Do. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids and the like, and organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically nontoxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, eye Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suve Latex, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobene Sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1 Sulfonates, naphthalene-2-sulfonates, mandelate and the like.

Acid addition salts according to another aspect of the present invention can be prepared by conventional methods, for example, by dissolving a derivative of formula (I) in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile and the like, and adding an organic or inorganic acid. The resulting precipitate may be prepared by filtration and drying, or the solvent and excess acid may be distilled under reduced pressure and dried to crystallize in an organic solvent.

Bases can also be used to make pharmaceutically acceptable metal salts. Alkali metal or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. Corresponding salts are also obtained by reacting alkali or alkaline earth metal salts with a suitable negative salt (eg silver nitrate).

Furthermore, in another aspect of the present invention, not only the compound represented by Formula (I) and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like that can be prepared therefrom are included.

In another aspect of the invention, as shown in Scheme 1 below,

Reacting the compound represented by the formula (II) with the compound represented by the formula (III) to prepare a compound represented by the formula (IV);

Preparing a compound represented by Formula V from the compound represented by Formula IV prepared in the above step;

Preparing a compound represented by Chemical Formula VI through a cyclization reaction through dehydration of the compound represented by Chemical Formula V prepared in the above step; And

There is provided a process for preparing a compound represented by the formula (I), which comprises the step of preparing a compound represented by the formula (I) by removing a protecting group from the compound represented by the formula (VI) prepared in the step.

Scheme 1

(In Scheme 1,

L is absent, -O-, -NH-, -NR-, or -S-;

n, R, R ¹ to R ⁵ are independently as defined in formula (I) above; And

PG ₁ to PG ₃ are protecting groups.

In another aspect of Scheme 1,

L is -NH-;

R ¹ and R ² are -NH ₂ ;

R ³ is as defined in Formula I above;

R ⁴ and R ⁵ are -OH;

PG ₁ is tert-Butyloxycarbonyl (Boc);

PG ₂ is acetoxy (OAc); And

PG ₃ is triphenylmethyl.

The reaction temperature of the step of preparing the compound represented by the formula (IV) by reacting the compound represented by the formula (II) with the compound represented by the formula (III) is not particularly limited, but may be performed at room temperature. In this case, the room temperature is a few examples of 10 ℃ to 50 ℃, 20 ℃ to 50 ℃, 25 ℃ to 50 ℃, 30 ℃ to 50 ℃, 35 ℃ to 50 ℃, 40 ℃ to 50 ℃, 45 ℃ to 50 ℃, 10 ° C to 40 ° C, 10 ° C to 35 ° C, 10 ° C to 30 ° C, 10 ° C to 25 ° C, 10 ° C to 20 ° C, or 10 ° C to 15 ° C. In addition, the reaction time is not particularly limited, but in some examples, 1h to 24h, 5h to 24h, 10h to 24h, 15h to 24h, 20h to 24h, 1h to 20h, 1h to 15h, 1h to 10h, or 1h to 5h. Furthermore, a target compound may be obtained through a work-up process generally performed in the field of synthesis.

From the compound represented by the formula (IV) prepared in the above step, the reaction temperature and reaction time of the step of preparing the compound represented by the formula (V) are the same as the above conditions and are omitted to avoid duplicate description. This step is to remove the protecting group, which may be removed through acid treatment, commonly known in the art of compound synthesis.

The reaction temperature and reaction time of the step of preparing the compound represented by the formula (VI) through the cyclization reaction through dehydration of the compound represented by the above formula (V) are the same as the above conditions, and thus will be omitted to avoid duplicate explanation. . This step is a step of cyclization through dehydration, which can induce cyclization through a dehydration reaction commonly known in the art of compound synthesis.

Removing the protecting group from the compound represented by the formula (VI) prepared in the above step, the reaction temperature and reaction time of the step of preparing the compound represented by the formula (I) are the same as the above conditions and are omitted to avoid redundant description. This step is to remove the protecting group, it is possible to remove the protecting group through the acid treatment commonly known in the art of compound synthesis, from which the amatoxin derivative provided in one aspect of the present invention can be prepared.

Hereinafter, a method of preparing a compound represented by Chemical Formula II and a compound represented by Chemical Formula III will be described.

The compound represented by Formula II is as shown in Scheme 2 below,

Reacting the compound represented by the formula (X1) with the compound represented by the formula (X2) to prepare a compound represented by the formula (X3);

Preparing a compound represented by Chemical Formula X4 by converting the -OMe group of the compound represented by Chemical Formula X3 prepared in the above step into -OH;

Preparing a compound represented by Chemical Formula X5 by cyclizing the compound represented by Chemical Formula X4 prepared in the above step; And

The -OtBu group of the compound represented by Formula X5 prepared in the above step may be converted to -OH to prepare a compound represented by Formula II.

Scheme 2

(In Scheme 2,

L is absent, -O-, -NH-, -NR-, or -S-;

n is as defined in formula (I) above; And

PG ₁ and PG ₄ are protecting groups.

On the other side,

PG ₁ is tert-Butyloxycarbonyl (Boc);

PG ₄ may be 9-Fluorenylmethyloxycarbonyl (Fmoc).

The reaction temperature and reaction time in the step of preparing the compound represented by the formula (X3) by reacting the compound represented by the formula (X1) with the compound represented by the formula (X3) are the same as the above conditions, and thus will be omitted to avoid redundant description. This step is a step in which the compound represented by the formula (X3) is prepared by combining the amine group of the compound represented by the formula (X1) and the dehydration reaction of the hydroxyl group of the compound represented by the formula (X2).

The reaction temperature of the step of preparing the compound represented by the formula (X4) by converting the -OMe group of the compound represented by the formula (X3) prepared in the above step into -OH is not particularly limited. ℃, 50 ℃ to 120 ℃, 60 ℃ to 120 ℃, 70 ℃ to 120 ℃, 80 ℃ to 120 ℃, 90 ℃ to 120 ℃, 100 ℃ to 120 ℃, 110 ℃ to 120 ℃, 40 ℃ to 110 ℃, 40 ° C to 100 ° C, 40 ° C to 90 ° C, 40 ° C to 80 ° C, 40 ° C to 70 ° C, 40 ° C to 60 ° C, or 40 ° C to 50 ° C. Since the reaction time is the same as the above condition, it is omitted to avoid redundant description. This step may use a known method of converting the -OMe group to -OH, in one embodiment can be carried out by treating Me ₃ SnOH to the compound represented by the formula (X3).

Since the reaction temperature of the step of preparing the compound represented by the formula (X5) by cyclization reaction of the compound represented by the formula (X4) prepared in the above step is the same as the above conditions, it will be omitted to avoid redundant description. However, the reaction time is 1 minutes to 20 minutes, 5 minutes to 20 minutes, 10 minutes to 20 minutes, 15 minutes to 20 minutes, 1 minute to 15 minutes, 1 minute to 10 minutes, or 1 minute to 5 by some examples. Can be done in minutes. In this step, the protecting group PG ₄ is removed and the resulting amine group is cyclized through a dehydration reaction with an adjacent hydroxyl group.

The reaction temperature of the step of preparing the compound represented by Chemical Formula II by converting the -OtBu group of the compound represented by Chemical Formula X5 prepared in the above step into -OH is not particularly limited, but may be performed at room temperature. In this case, the room temperature is a few examples of 10 ℃ to 50 ℃, 20 ℃ to 50 ℃, 25 ℃ to 50 ℃, 30 ℃ to 50 ℃, 35 ℃ to 50 ℃, 40 ℃ to 50 ℃, 45 ℃ to 50 ℃, 10 ° C to 40 ° C, 10 ° C to 35 ° C, 10 ° C to 30 ° C, 10 ° C to 25 ° C, 10 ° C to 20 ° C, or 10 ° C to 15 ° C. In addition, the reaction time is not particularly limited, but in some examples, 1h to 24h, 5h to 24h, 10h to 24h, 15h to 24h, 20h to 24h, 1h to 20h, 1h to 15h, 1h to 10h, or 1h to 5h. This step is a step of converting a -OtBu group to -OH, can be used without limitation if the conversion method generally known in the field of compound synthesis, one example may be carried out by treating HCl dissolved in dioxane.

Compound represented by Formula III is shown in Scheme 3 below,

Reacting the compound represented by the formula (Y1) with the compound represented by the formula (Y2) to prepare a compound represented by the formula (Y3); And

It may be prepared by removing the PG ₄ protecting group of the compound represented by the formula Y3 prepared in the above step, to prepare a compound represented by the formula (III).

Scheme 3

(In Scheme 3,

R ³ is as defined in Formula I above; And

PG ₂ to PG ₄ are protecting groups).

On the other side,

PG ₂ is acetoxy (OAc);

PG ₃ is triphenylmethyl; And

PG ₄ may be 9-Fluorenylmethyloxycarbonyl (Fmoc).

The reaction temperature of the step of preparing the compound represented by the formula Y3 by reacting the compound represented by the formula Y1 with the compound represented by the formula Y2 is not particularly limited, but may be performed at room temperature. In this case, the room temperature is a few examples of 10 ℃ to 50 ℃, 20 ℃ to 50 ℃, 25 ℃ to 50 ℃, 30 ℃ to 50 ℃, 35 ℃ to 50 ℃, 40 ℃ to 50 ℃, 45 ℃ to 50 ℃, 10 ° C to 40 ° C, 10 ° C to 35 ° C, 10 ° C to 30 ° C, 10 ° C to 25 ° C, 10 ° C to 20 ° C, or 10 ° C to 15 ° C. In addition, the reaction time is not particularly limited, but in some examples, 1h to 24h, 5h to 24h, 10h to 24h, 15h to 24h, 20h to 24h, 1h to 20h, 1h to 15h, 1h to 10h, or 1h to 5h. This step is a reaction in which the hydroxyl group of the compound represented by the formula (Y1) and the amine group of the compound represented by the formula (Y2) are bonded through dehydration condensation.

Removing the PG ₄ protecting group of the compound represented by the formula Y3 prepared in the above step, the reaction temperature of the step of preparing a compound represented by the formula (III) is the same as the above conditions are omitted to avoid duplicate explanation. However, the reaction time is 1 minutes to 60 minutes, 10 minutes to 60 minutes, 20 minutes to 60 minutes, 30 minutes to 60 minutes, 40 minutes to 60 minutes, 50 minutes to 60 minutes, 1 minute to 50 minutes by some examples. , 1 minute to 40 minutes, 1 minute to 30 minutes, 1 minute to 20 minutes, or 1 minute to 10 minutes.

In one aspect of the invention,

There is provided a compound represented by the following formula α, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, which is a synthetic intermediate.

[Formula α]

(In the above formula α,

n and L are as defined in Scheme 1 above; And

A ¹ and A ² are independently —H, or a protecting group).

On the other side,

The protecting groups of A ¹ and A ² may be tert-butyloxycarbonyl (Boc).

In another aspect,

The present invention provides a compound represented by the following formula β, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, which is a synthetic intermediate.

[Formula β]

(In the above formula β,

R ³ is as defined in Formula I above; And

E ¹ , E ² and E ³ are independently —H, —OH, or a protecting group).

On the other side,

The protecting group of E ¹ , E ² is acetoxy (OAc); And

The protecting group of E ³ may be triphenylmethyl.

In another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating cancer containing the compound represented by the formula (I), the optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. In another aspect, the pharmaceutical composition may be in the form of ADC.

In another aspect of the present invention, the compound represented by Formula (I) may be administered in various oral and parenteral formulations during clinical administration, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, It is prepared using diluents or excipients such as surfactants.

Solid form preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, which form at least one excipient such as starch, calcium carbonate, water, or the like. It is prepared by mixing cross, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, or syrups, and include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Can be.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

In addition, in another aspect of the invention, the effective dosage of the compound to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and generally about 0.001-100 mg / kg / Day, preferably 0.01 to 35 mg / kg / day. Based on an adult patient with a weight of 70 kg, it is generally 0.07 ~ 7000 mg / day, preferably 0.7 ~ 2500 mg / day, once a day at regular intervals depending on the judgment of the doctor or pharmacist Multiple doses may be administered.

Hereinafter, the content according to an aspect of the present invention will be described in detail through examples and experimental examples. However, Examples and Experimental Examples described below are merely to illustrate the invention, but the present invention is not limited thereto.

Example 1 (2R, 3R, 4S) -5- (tert-butoxy) -4-((2S, 4R) -4-hydroxy-1- (N ⁴ -Trityl-L-asparaginyl) pyrrolidine-2-carboxamido) -3-methyl-5-oxopentan-1,2-diyl diacetate (1-1)

Step 1: (2S, 4R) -2-((allyloxy) carbonyl) -4-hydroxypyrrolidine-1-ium 4-methylbenzenesulfonate (1a)

In a dried flask with autostirring bar and Dean-Stark receiver, trans-L-hydroxyproline (1.0 g, 7.63 mmol), p-toluene sulfonic acid monohydrate (1.6 g, 8.39 mmol), allyl alcohol (5.2 mL, 76.3 mmol) and benzene (60 mL). The mixture was heated to reflux for 4 h to obtain a brown homogeneous solution. The solution was cooled to room temperature and concentrated under reduced pressure to afford the title salt 1a (2.52 g, 7.34 mmol, 96%) as a brown oil. The crude mixture was used for next step without further purification.

Step 2: Allyl- (2S, 4R) -1- (N ² -(((9H-fluoren-9-yl) methoxy) carbonyl) -N ⁴ -Trityl-L-asparaginyl) -4-hydroxypyrrolidine-2-carboxylate (1b)

The trityl -L- asparagine (1.0 g, 1.67 mmol) solution, - THF N ² dissolved in (25 mL) - ((( 9 H - fluoren-9-yl) methoxy) carbonyl) - N ⁴ 1a (1.15 g, 3.35 mmol) prepared in step 1 was added. HATU (1.27 g, 3.35 mmol) and DIPEA (0.87 mL, 5.02 mmol) were added sequentially at 0 ° C. and the reaction mixture was stirred at rt for 12 h. TLC analysis confirmed complete consumption of starting material. The reaction mixture is concentrated to remove THF, water is added, then concentrated with EtOAc (2 X 50 mL), the organic layer is washed with brine solution (2 X 15 mL), dried over sodium sulfate, Concentration under reduced pressure gave the crude product which was purified by column chromatography to give the title compound 1b (810 mg, 1.08 mmol, 64%) as a white solid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.72 (d, J = 7.6 Hz, 2H), 7.52 (t, J = 6.5 Hz, 2H), 7.36 (t, J = 7.4 Hz, 2H), 7.39-7.21 (m, 16H), 7.14 (dd, J = 8.0, 1.7 Hz, 1H), 6.31 (d, J = 7.89 Hz, 1H), 5.88-5.76 (m, 1H), 5.30-5.17 (m, 2H), 4.76 (d, J = 6.5 Hz, 1H), 4.66-4.60 (m, 1H), 4.53-4.44 (m, 3H), 4.32-4.11 (m, 1H), 3.76-3.59 (m, 2H), 2.69 ( s, 2H), 2.50 (s, 1 H), 2.02-1.93 (m, 1 H);

¹³ C NMR (125 MHz, CD ₃ OD) δ 173.00, 172.96, 172.65, 170.96, 158.00, 145.81, 145.74, 145.18, 145.06, 142.53, 142.50, 133.29, 130.05, 128.77, 128.74, 128.71, 128.20, 128.18, 127 126.26, 126.21, 126.10, 120.89, 118.76, 71.91, 70.86, 68.19, 66.82, 61.51, 59.56, 56.12, 51.60, 48.25, 39.56, 38.19, 20.86, 14.46;

LC / MS mlz 750.7 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₄₆ H ₄₃ N ₃ O ₇ [M ⁺ ] 749.3101.

Step 3: (2S, 4R) -1- (N ² -(((9H-fluoren-9-yl) methoxy) carbonyl) -N ⁴ -Trityl-L-asparaginyl) -4-hydroxypyrrolidine-2-carboxylic acid (1c)

To the solution of 1b (800 mg, 1.06 mmol) compound of Step 2 above dissolved in THF (200 mL), PhSiH ₃ (577 mg, 5.33 mmol) was added. 0, under a nitrogen atmosphere, Pd (PPh ₃ ) ₄ (123 mg, 0.106 mmol) was added to the mixture and stirred at rt for 4 h. Then diluted with H ₂ O and extracted with EtOAc (2 × 150 mL). The collected organic layer was distilled under reduced pressure to give a crude mixture, which was purified by silica gel column chromatography using MeOH / MC (1/4) to give the title compound 1c (530 mg, 0.746 mmol, 70%) as an off-white solid. It was.

¹ H NMR (300 MHz, DMSO- d ₆ ) δ 8.58 (s, 1H), 7.89 (d, J = 7.6 Hz, 2H), 7.77-7.62 (m, 3H), 7.41 (d, J = 7.0 Hz, 2H), 7.33-7.07 (m, 18H), 5.17 (s, 1H), 4.49 (t, J = 8.1 Hz, 1H), 4.41-4.12 (m, 4H), 3.58 (dd, J = 10.4, 4.7 Hz , 1H), 3.49-3.38 (m, 1H), 2.62 (dd, J = 14.6, 9.9 Hz, 1H), 2.47-2.36 (m, 1H), 2.15-1.98 (m, 1H), 1.96-1.82 (m , 1H);

¹³ C NMR (125 MHz, CD ₃ OD) δ 175.15, 172.65, 171.06, 158.03, 145.82, 145.73, 145.17, 145.08, 142.51, 133.09, 133.00, 130.05, 129.90, 128.76, 128.74, 128.70, 128.21, 128.18, 1287918 126.20, 126.10, 120.88, 71.90, 70.86, 68.19, 61.52, 59.45, 56.08, 51.67, 48.26, 39.59, 38.32, 20.86, 14.46;

LC / MS mlz 708.2 [M-H ⁺ ].

HRMS (EI) m / z calcd for C ₄₃ H ₃₉ N ₃ O ₇ [M ⁺ ] 709.2788.

Step 4: (2R, 3R, 4S) -4-((2S, 4R) -1- (N ² -(((9H-fluoren-9-yl) methoxy) carbonyl) -N ⁴ -Trityl-L-asparaginyl) -4-hydroxypyrrolidine-2-carboxamido) -5- (tert-butoxy) -3-methyl-5-oxopentan-1,2-di Diacetate (1)

To the 1c (500 mg, 0.704 mmol) compound solution of step 3 dissolved in DCM (25 mL) at 0 ° C., the compound represented by formula II-11 (263 mg, 0.774 mmol) in the above scheme was added. To this mixed solution, PyBOP (476 mg, 0.915 mmol) and DIPEA (0.36 mL, 2.11 mmol) were added and stirred for 12 h. The reaction mixture was quenched with water (10 mL) and extracted with DCM (2 X 25 mL). The collected organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure to give crude product. The crude product was purified via column chromatography using MeOH / MC (1: 9) as eluent to afford the title compound 1 (595 mg, 0.598 mmol, 85%) as a white solid.

¹ H NMR (300 MHz, CD3OD) δ 7.80 (d, J = 7.5 Hz, 2H), 7.69-7.62 (m, 2H), 7.39 (t, J = 7.5 Hz, 2H), 7.26 (q, J = 6.0 Hz, 17H), 5.04-4.95 (m, 1H), 4.73-4.56 (m, 2H), 4.46-4.30 (m, 5H), 4.26-4.18 (m, 1H), 3.99 (dd, J = 12.4, 5.8 Hz, 1H), 3.78-3.66 (m, 1H), 3.53 (dd, J = 10.8, 4.4 Hz, 1H), 3.23 (q, J = 7.4 Hz, 1H), 2.73 (d, J = 7.0 Hz, 2H ), 2.36-2.16 (m, 2H), 2.12-2.06 (m, 1H), 2.04-2.00 (m, 3H), 1.96 (s, 2H), 1.47 (s, 9H), 0.97 (d, J = 7.0 Hz, 2H), 0.83 (d, J = 7.0 Hz, 1H).

¹³ C NMR (125 MHz, CDCl 3) δ 157.29, 155.37, 155.25, 146.54, 138.35, 134.64, 131.35, 128.68, 127.90, 125.01, 123.59, 123.33, 121.17, 120.62, 117.84, 110.48, 106.49, 56.02, 56.02 18.94, 15.37.

LC / MS mlz 995.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₅₇ H ₆₂ N ₄ O ₁₂ [M ⁺ ] 994.4364.

Step 5: (2R, 3R, 4S) -5- (tert-butoxy) -4-((2S, 4R) -4-hydroxy-1- (N ⁴ -Trityl-L-asparaginyl) pyrrolidine-2-carboxamido) -3-methyl-5-oxopentan-1,2-diyl diacetate (1-1)

To 1 (590 mg, 0.593 mmol) compound solution prepared in step 4, dissolved in DCM (15 mL) at rt, DBU (95 mg, 0.622 mmol) was added and stirred for 30 minutes. TLC analysis confirmed complete consumption of starting material. The reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (2 X 30 mL). The collected organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure to give crude product. The crude product was purified via column chromatography using MeOH / MC (1: 4) as eluent to afford the title compound 1-1 (385 mg, 0.498 mmol, 84%) as a white solid.

¹ H NMR (300 MHz, CD3OD) δ 7.28-7.04 (m, 15H), 4.94-4.82 (m, 1H), 4.50-4.41 (m, 1H), 4.34-4.29 (m, 1H), 4.28-4.23 ( m, 1H), 4.18 (dd, J = 12.3, 2.5 Hz, 1H), 3.92-3.82 (m, 1H), 3.81-3.75 (m, 1H), 3.38-3.31 (m, 1H), 3.27-3.23 ( m, 1H), 2.65-2.45 (m, 2H), 2.21-2.08 (m, 2H), 1.99-1.93 (m, 1H), 1.92-1.89 (m, 3H), 1.85 (s, 2H), 1.41- 1.32 (m, 9H), 0.84 (d, J = 7.1 Hz, 2H), 0.69 (d, J = 7.0 Hz, 1H).

¹³ C NMR (125 MHz, CDCl 3) δ 157.29, 155.37, 155.25, 146.54, 138.35, 134.64, 131.35, 128.68, 127.90, 125.01, 123.59, 123.33, 121.17, 120.62, 117.84, 110.48, 106.49, 56.02, 56.02 18.94, 15.37.

LC / MS mlz 773.3 [M + H < + >].

HRMS (EI) m / z calcd for C ₄₂ H ₅₂ N ₄ O ₁₀ [M ⁺ ] 772.3683, found 772.3672.

Example 2 tert-Butyl S- (6-((tert-butoxycarbonyl) amino) -3-((S) -2-((tert-butoxycarbonyl) amino) -3-methoxy -3-oxopropyl) -1H-indol-2-yl) -L-cysteinate (2-1)

Step 1: (S) -2-amino-3- (6-nitro-1H-indol-3-yl) propanoic acid (2a)

To a suspension of L-tryptophan (20.4 g, 100 mmol) and urea (0.25 g, 4.16 mmol) dissolved in glacial acetic acid (250 mL), a fuming nitric acid solution dissolved in glacial acetic acid (15 mL) was added. The resulting yellow solution was stirred at 0 until it turned into a suspension. Fuming nitric acid (8.75 mL) dissolved in glacial acetic acid (35 mL) was added slowly at 15 ° C. as a droplet. After stirring for 18 h at Rt, the resulting yellow precipitate was collected by filtration to give the crude product, which was dissolved in hot water (100 mL), neutralized with sodium carbonate and recrystallized with ethanol to give the title compound 2a (10.7 g, 42.9 mmol, 43%) The compound was prepared as a yellow solid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 8.45 (s, 1H), 8.04 (d, J = 9.0 Hz, 1H), 7.83 (d, J = 9.0 Hz, 1H), 7.67 (s, 1H), 3.69 (t, J = 6.0 Hz, 1H), 3.18-3.33 (m, 2H);

LC / MS mlz 249.1 [M + H ⁺ ].

Step 2: methyl (S) -2-amino-3- (6-nitro-1H-indol-3-yl) propanoate hydrochloride (2b)

The solution of 2a (10.0 g, 40.1 mmol) compound of step 1 dissolved in MeOH (80 mL) was cooled to 0 ° C., thionyl chloride (6.4 mL, 88.2 mmol) was added and stirred for 30 minutes. The reaction mixture was heated to reflux and stirred for 8 h. TLC analysis confirmed complete consumption of starting material. The reaction mixture was concentrated under reduced pressure to remove thionyl chloride and MeOH. DCM was added to the residue, the solid precipitate was filtered and dried to give the title compound 2b (11.3 g, 37.70 mmol, 94%) as a white solid.

¹ H NMR (300 MHz, CD ₃ OD) δ 8.39 (d, J = 1.9 Hz, 1H), 8.01 (dd, J = 8.9, 2.1 Hz, 1H), 7.69 (d, J = 8.8 Hz, 1H), 7.57 (s, 1 H), 4.38 (dd, J = 6.9, 6.0 Hz, 1 H), 3.80 (s, 3 H), 3.54-3.34 (m, 2H);

¹³ C NMR (125 MHz, DMSO) δ 169.53, 141.93, 134.59, 132.15, 131.82, 118.44, 113.82, 108.41, 108.09, 52.72, 52.61, 25.57;

LC / MS mlz 264.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₁₂ H ₁₃ N ₃ O ₄ [M ⁺ ] 263.0906.

Step 3: methyl (S) -2-amino-3- (6-amino-1 H-indol-3-yl) propanoate hydrochloride (2c)

Pd / C to 2b (10 g, 33.3 mmol) compound solution prepared in step 2 dissolved in MeOH (100 ml) (10 mol%, 1.0 g) was added. The mixture was stirred for 12 h under a hydrogen atmosphere (balloon). After confirming that the reaction was completed by TLC, the catalyst was removed by a filter through celite. The filtered mixture was concentrated under reduced pressure to afford the title compound 2c (8.9 g, 32.9 mmol, 99%) as a brown solid.

¹ H NMR (300 MHz, CD ₃ OD) δ 7.32 (d, J = 8.4 Hz, 1H), 6.98 (s, 1H), 6.82 (d, J = 1.8 Hz, 1H), 6.65 (dd, J = 8.4 , 1.9 Hz, 1H), 4.24 (dd, J = 7.5, 5.4 Hz, 1H), 3.79 (s, 3H), 3.41-3.20 (m, 2H);

¹³ C NMR (125 MHz, CD ₃ OD) δ 171.32, 141.71, 139.31, 123.96, 122.69, 119.48, 112.34, 107.75, 99.83, 54.70, 53.52, 27.93;

LC / MS mlz 234.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₁₂ H ₁₅ N ₃ O ₂ [M ⁺ ] 233.1164, found 233.1164.

Step 4: methyl- (S) -2-((tert-butoxycarbonyl) amino) -3- (6-((tert-butoxycarbonyl) amino) -1H-indol-3-yl) propano Eight (2d)

To a solution of 2c (5.0 g, 18.53 mmol) compound dissolved in DCM (150 mL) at 0 ° C., Boc anhydride (8.9 g, 40.78 mmol) was added and triethylamine (8.3 mL, 59.3 mmol) was added. The reaction mixture was stirred at rt for 5 h. Upon completion the reaction was quenched with water (100 mL) and extracted with DCM (2 X 150 mL). Sat organic layer collected. Washed with brine solution, dried over Na ₂ SO ₄ , and the solvent was removed under reduced pressure to give crude product, which was purified by column chromatography to give the title compound 2d (7.9 g, 18.22 mmol, 98%) as white Prepared as a solid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.95 (s, 1H), 7.76 (s, 1H), 7.41 (d, J = 8.5 Hz, 1H), 6.93 (d, J = 1.6 Hz, 1H), 6.78 (dd, J = 8.9, 1.3 Hz, 1H), 6.51 (s, 1H), 5.05 (d, J = 7.5 Hz, 1H), 4.69-4.53 (m, 1H), 3.66 (s, 3H), 3.24 ( d, J = 5.4 Hz, 2H), 1.53 (s, 9H), 1.42 (s, 9H);

¹³ C NMR (125 MHz, CDCl ₃ ) δ 172.93, 155.37, 153.35, 136.70, 133.44, 123.97, 122.72, 118.87, 112.52, 109.89, 101.81, 80.30, 79.94, 54.29, 52.31, 28.53, 28.44, 28.14;

LC / MS mlz 434.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₂₂ H ₃₁ N ₃ O ₆ [M ⁺ ] 433.2213, found 433.2213.

Step 5: di-tert-butyl-3,3'-disulfanediyl (2R, 2'R) -bis (2-((((9H-fluoren-9-yl) methoxy) carbonyl) amino Propanoate) (2e)

(2R, 2'R) -3,3'-disulfanediylbis (1- (tert-butoxy) -1-oxopropane dissolved in water (25 mL) and dioxane (50 mL) at 0 ° C. To a solution of -2-aluminum) chloride (5.0 g, 11.75 mmol), NaHCO 3 (4.9 g, 58.76 mmol) and Fmoc-OSu (11.9 g, 35.2 mmol) were added sequentially. The reaction mixture was stirred at rt for 12 h. At the end of the reaction, the reaction mixture was extracted with DCM (2 X 150 mL). Sat organic layer collected. Washed with brine solution, dried over Na ₂ SO _4, and the solvent was removed under reduced pressure to afford the crude product, which was purified by recrystallization with DCM / EtOAc to give the title compound 2e (8.2 g, 10.28 mmol, 87% ) Was prepared as a white solid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.77 (d, J = 7.3 Hz, 4H), 7.61 (d, J = 7.2 Hz, 4H), 7.40 (t, J = 7.5 Hz, 4H), 7.32 (d , J = 7.2 Hz, 4H), 5.75 (d, J = 8.1 Hz, 2H), 4.60 (d, J = 7.7 Hz, 2H), 4.38 (d, J = 6.6 Hz, 4H), 4.22 (t, J = 7.2 Hz, 2H), 3.23 (t, J = 5.8 Hz, 4H), 1.50 (s, 18H);

¹³ C NMR (125 MHz, CDCl ₃ ) δ 169.45, 155.83, 143.94, 143.89, 141.39, 127.81, 127.18, 125.28, 120.08, 83.25, 67.35, 54.28, 47.22, 42.03, 28.13;

LC / MS mlz 434.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₄₄ H ₄₈ N ₂ O ₈ S ₂ [M ⁺ ] 796.2852, found 796.2831.

Step 6: tert-butyl N-(((9H-fluoren-9-yl) methoxy) carbonyl) -S- (6-((tert-butoxycarbonyl) amino) -3-((S) -2-((tert-butoxycarbonyl) amino) -3-methoxy-3-oxopropyl) -1 H-indol-2-yl) -L-cysteinate (2)

The solution of 2e (8.0 g, 10.0 mmol) compound of step 5 dissolved in anhydrous chloroform (90 mL) was vigorously stirred in the atmosphere, and sulfarylyl chloride (3.24 mL, 40.1 mmol) was added dropwise. The solution was stirred for 45 minutes at room temperature under air, and it was found to change to yellow over time. TLC confirmed that the production of sulfenyl chloride was mostly complete. The solution was quickly concentrated in vacuo to remove excess sulfyl chloride to give sulfenyl chloride as a viscous yellow oil. The sulfenyl chloride was immediately dissolved in anhydrous CHCl 3 and THF (3: 1) (120 mL) and stirred for 5 minutes in an ice bath under a nitrogen atmosphere. The solution was rapidly treated with NaHCO 3 (2.81 g, 33.4 mmol) and immediately dissolved in anhydrous CHCl 3 and THF (3: 1) (25 mL) solution in which the compound represented by the formula 2d (2.9 g, 6.69 mmol) was dissolved in the scheme. Was treated. The solution was stirred in an ice bath for 15 minutes, warmed to room temperature and stirred for 12 h. The solution was concentrated under reduced pressure and the residue was purified by silica flash column chromatography using EA / MC (1: 9) as eluent to afford the title compound 2 (3.5 g, 4.21 mmol, 63%) as a white solid. Prepared as.

¹ H NMR (300 MHz, CDCl ₃ ) δ 9.84 (s, 1H), 7.77 (d, J = 7.7 Hz, 2H), 7.64 (d, J = 6.6 Hz, 2H), 7.44-7.27 (m, 6H) , 6.75 (d, J = 9.7 Hz, 1H), 6.49 (s, 1H), 5.59 (d, J = 8.1 Hz, 1H), 5.12 (d, J = 9.9 Hz, 1H), 4.59 (d, J = 5.9 Hz, 1H), 4.53-4.35 (m, 3H), 4.24 (t, J = 6.8 Hz, 1H), 3.58 (s, 3H), 3.32 (d, J = 6.3 Hz, 2H), 2.73-2.64 ( m, 1H), 1.52 (s, 9H), 1.44 (s, 9H);

¹³ C NMR (125 MHz, CDCl ₃ ) δ 172.79, 169.74, 156.92, 155.19, 153.01, 143.88, 143.80, 141.44, 141.40, 137.31, 137.16, 134.39, 127.82, 127.21, 125.41, 125.30, 125.02, 124.02, 124.02, 124.02 , 119.41, 115.87, 112.46, 101.12, 83.59, 80.40, 79.84, 67.60, 54.70, 54.61, 54.26, 54.17, 52.36, 47.21, 40.97, 28.52, 28.48, 28.15, 28.07;

LC / MS mlz 831.4 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₄₄ H ₅₄ N ₄ O ₁₀ S [M ⁺ ] 830.3561.

Step 7: tert-Butyl S- (6-((tert-butoxycarbonyl) amino) -3-((S) -2-((tert-butoxycarbonyl) amino) -3-methoxy-3 -Oxopropyl) -1H-indol-2-yl) -L-cysteinate (2-1)

To 2 (3.5 g, 4.211 mmol) compound solution of step 6 dissolved in DCM (250 mL) at rt, DBU (0.67 g, 4.42 mmol) was added dropwise and the reaction mixture was stirred for 1 h. TLC analysis confirmed complete consumption of starting material. The reaction mixture was quenched with water (100 mL) and extracted with DCM (2 X 150 mL). The collected organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure to give the crude product. The crude product was purified via column chromatography using MeOH / MC (1: 4) as eluent to afford the title compound 2-1 (2.2 g, 3.61 mmol, 86%) as a brown solid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 10.64 (s, 1H), 7.74 (s, 1H), 7.33 (d, J = 8.5 Hz, 1H), 6.70 (d, J = 8.3 Hz, 1H), 6.53 (s, 1H), 5.15 (d, J = 8.0 Hz, 1H), 4.57 (q, J = 6.5 Hz, 1H), 3.72 (t, J = 5.1 Hz, 1H), 3.65 (s, 3H), 3.26 (d, J = 5.5 Hz, 2H), 3.04 (d, J = 4.3 Hz, 2H), 1.83 (s, 2H), 1.53 (s, 9H), 1.49 (s, 9H), 1.40 (s, 9H) ;

¹³ C NMR (125 MHz, CD ₃ OD) δ 174.41, 173.84, 157.51, 155.59, 138.89, 136.02, 126.04, 125.15, 119.74, 116.77, 82.93, 80.61, 56.08, 55.76, 52.68, 41.78, 28.79, 28.71, 28.71, 28.71 28.20;

LC / MS mlz 608.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₂₉ H ₄₄ N ₄ O ₈ S [M ⁺ ] 608.2880.

Example 3 (((9H-Fluoren-9-yl) methoxy) carbonyl) glycyl-L-isosilylglycine

Step 1: 2- (allyloxy) -2-oxoethane-1-aluminum 4-methylbenzenesulfonate (3a)

In a dried flask with autostirring bar and Dean-Stark receiver, glycine (2.0 g, 26.3 mmol), p-toluene sulfonic acid monohydrate (5.0 g, 26.3 mmol), allyl alcohol (10.7 mL, 158 mmol) and Benzene (100 mL) was added. The mixture was heated to reflux for 4 h until 9.0 ml H 2 O was collected and complete conversion was confirmed by TLC. The suspension was concentrated under reduced pressure and recrystallized with ether (100 mL) to give the title salt 3a (7.3 g, 25.4 mmol, 97%) compound as a white solid.

Step 2: allyl (((9H-fluoren-9-yl) methoxy) carbonyl) -L-isosilylglycinate (3b)

To a solution of Fmoc-L-isoleucine (4.0 g, 11.3 mmol) dissolved in DCM (40 mL) at 0 ° C., 3a (3.9 g, 13.6 mmol) compound prepared in step 1 above was added. PyBOP (7.64 g, 14.7 mmol) and DIPEA (5.9 mL, 33.9 mmol) were added to the reaction mixture, and the reaction mixture was stirred for 2 h. The reaction mixture was quenched with water (40 mL) and extracted with DCM (2 X 45 mL). The collected organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure to give the crude product. The crude product was purified via column chromatography using EtOAc / Hx (1: 1) as eluent to afford the title compound 3b (4.6 g, 10.2 mmol, 90%) as a white solid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.77 (d, J = 7.5 Hz, 2H), 7.59 (d, J = 7.2 Hz, 2H), 7.40 (t, J = 7.3 Hz, 2H), 7.31 (t , J = 7.3 Hz, 2H), 6.34 (s, 1H), 5.98-5.81 (m, 1H), 5.41-5.19 (m, 3H), 4.65 (d, J = 5.8 Hz, 2H), 4.42 (s, 2H), 4.22 (t, J = 6.8 Hz, 1H), 4.14-3.95 (m, 3H), 1.92 (s, 1H), 1.58 (s, 1H), 1.16 (s, 1H), 1.01-0.79 (m , 6H);

¹³ C NMR (125 MHz, CDCl ₃ ) δ 171.68, 169.37, 156.46, 143.94, 143.87, 141.42, 131.51, 127.83, 127.20, 127.17, 125.19, 120.10, 119.19, 67.16, 66.20, 59.72, 47.32, 41.35, 37.39. , 26.33, 24.88, 15.54, 11.48;

LC / MS mlz 451.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₂₆ H ₃₀ N ₂ O ₅ [M ⁺ ] 450.2155.

Step 3: Allyl (((9H-Fluoren-9-yl) methoxy) carbonyl) gylsil-L-isosilylglycinate (3)

To rt 3b (4.51 g, 10.0 mmol) compound solution of step 2 dissolved in DMF (70 mL) at rt, DBU (1.57 mL, 10.51 mmol) was added and the reaction mixture was stirred for 10 minutes. TLC analysis confirmed complete consumption of starting material. To this reaction mixture, a solution of Fmoc-Gly-OH (3.57 g, 12.01 mmol) and PyBOP (6.25 g, 12.01 mmol) dissolved in DMF (30 mL) was added and the reaction mixture was stirred for 1 h. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 X 250 mL). The collected organic layer was washed with brine solution, dried over sodium sulfate, and then concentrated under reduced pressure to obtain a crude product. The crude product was purified via silica gel column chromatography using EtOAc / nucleic acid (4: 1) as eluent to afford the title compound 3 (3.6 g, 7.09 mmol, 71%) as a white solid.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 8.51 (t, J = 5.9 Hz, 1H), 7.90 (d, J = 7.5 Hz, 2H), 7.85 (d, J = 8.9 Hz, 1H), 7.72 (d, J = 7.4 Hz, 2H), 7.55 (t, J = 6.1 Hz, 1H), 7.42 (t, J = 7.4 Hz, 2H), 7.33 (t, J = 7.4 Hz, 2H), 5.99-5.80 (m, 1H), 5.32 (d, J = 17.3 Hz, 1H), 5.22 (d, J = 10.5 Hz, 1H), 4.57 (d, J = 5.5 Hz, 2H), 4.26 (q, J = 7.0, 6.3 Hz, 4H), 4.00-3.75 (m, 2H), 3.68 (d, J = 6.1 Hz, 2H), 1.82-1.65 (m, 1H), 1.44 (s, 1H), 1.08 (dt, J = 14.6 , 7.6 Hz, 1H), 0.85 (d, J = 6.8 Hz, 3H), 0.80 (t, J = 7.3 Hz, 3H);

¹³ C NMR (125 MHz, DMSO) δ 171.50, 169.33, 168.88, 156.47, 143.84, 140.72, 132.35, 127.62, 127.08, 125.24, 120.10, 117.89, 65.74, 64.77, 56.53, 46.64, 43.31, 40.62, 36.95, 24.62 15.21, 11.12;

LC / MS mlz 508.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₂₈ H ₃₃ N ₃ O ₆ [M ⁺ ] 507.2369.

Step 4: (((9H-Fluoren-9-yl) methoxy) carbonyl) gylsil-L-isoleucineglycine (3-1)

At 0 ° C., to a suspension of 3 (3.5 g, 6.89 mmol) compound prepared in step 3, dissolved in THF (300 mL), PhSiH ₃ (4.25 mL, 34.4 mmol) was added. Pd (PPh ₃ ) ₄ (797 mg, 0.69 mmol) was added to the mixture and stirred for 2 h. The reaction mixture was diluted with H ₂ O and extracted with EtOAc (2 × 150 mL). The collected organic layer was distilled under reduced pressure to give crude product, which was purified by silica gel column chromatography using MeOH / MC (1/4) to give the title compound 3-1 (2.45 g, 5.24 mmol, 76%). The compound was prepared as a white solid.

¹ H NMR (500 MHz, DMSO-d6) δ 8.32 (t, J = 5.9 Hz, 1H), 7.89 (d, J = 7.4 Hz, 2H), 7.80 (d, J = 8.9 Hz, 1H), 7.71 ( d, J = 7.3 Hz, 2H), 7.53 (t, J = 6.0 Hz, 1H), 7.41 (t, J = 7.4 Hz, 2H), 7.32 (t, J = 7.3 Hz, 2H), 4.28-4.22 ( m, 4H), 3.76-3.67 (m, 4H), 1.78-1.68 (m, 1H), 1.48-1.39 (m, 1H), 1.11-0.98 (m, 1H), 0.84 (d, J = 6.8 Hz, 3H), 0.79 (t, J = 7.3 Hz, 3H);

¹³ C NMR (125 MHz, DMSO) δ 170.96, 168.91, 156.52, 143.87, 140.75, 133.74, 131.23, 127.67, 127.43, 127.33, 127.13, 125.29, 120.14, 65.80, 56.78, 46.67, 43.39, 37.03, 24.24, 24.24 11.20;

LC / MS mlz 468.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₂₅ H ₂₉ N ₃ O ₆ [M ⁺ ] 467.2056.

Example 4 Compound Represented by Formula 10

Step 1: tert-Butyl-N-(((9H-fluoren-9-yl) methoxy) carbonyl) gylsil-L-isosilylglycyl-S- (6-((tert-butoxycarbonyl ) Amino) -3-((S) -2-((tert-butoxycarbonyl) amino) -3-methoxy-3-oxopropyl) -1H-indol-2-yl) -L-cysteinate (4)

3-1 (1.7 g, 3.62 mmol) compound prepared in Example 3 was prepared in 2-1 (2.1 g, 3.45 mmol) compound solution prepared in Example 2, dissolved in DMF (50 mL) at rt. Added. PyBOP (2.7 g, 5.17 mmol) and DIPEA (0.9 mL, 1.848 mmol) were added to the reaction mixture and stirred for 12 h. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 X 250 mL). The collected organic layer was washed with brine solution, dried over sodium sulfate, and then concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using MeOH / MC (1: 4) as eluent to afford the title compound 4 (3.1 g, 2.93 mmol, 85%) as a brown solid.

¹ H NMR (300 MHz, CD3OD) δ 7.77 (d, J = 7.5 Hz, 2H), 7.68 (s, 1H), 7.66-7.55 (m, 1H), 7.37 (t, J = 7.1 Hz, 3H), 7.28 (t, J = 7.3 Hz, 2H), 6.92 (d, J = 8.4 Hz, 1H), 4.47-4.34 (m, 1H), 4.31 (d, J = 6.9 Hz, 3H), 4.25-4.11 (m , 2H), 4.05-3.95 (m, 1H), 3.87-3.73 (m, 3H), 3.65-3.54 (m, 3H), 3.41-3.36 (m, 1H), 3.30-3.24 (m, 1H), 3.21 -3.11 (m, 1H), 3.07-2.95 (m, 0H), 2.02-1.85 (m, 1H), 1.68-1.57 (m, 1H), 1.53 (s, 9H), 1.39 (s, 9H), 1.36 (s, 9H), 1.28-1.21 (m, 1H), 0.99 (d, J = 6.8 Hz, 3H), 0.92 (t, J = 7.3 Hz, 3H);

¹³ C NMR (125 MHz, CD ₃ OD) δ 174.46, 174.40, 172.57, 171.50, 170.70, 158.98, 157.49, 155.62, 145.19, 142.52, 138.78, 136.06, 133.09, 130.00, 129.91, 128.77, 128.17, 126.22, 126.22 120.91, 119.92, 117.53, 83.61, 80.62, 68.28, 60.01, 56.11, 54.46, 52.70, 48.26, 47.34, 44.78, 43.48, 38.61, 37.57, 28.81, 28.68, 28.16, 26.10, 15.91, 11.49;

LC / MS mlz 1058.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₅₄ H ₇₁ N ₇ O ₁₃ S [M ⁺ ] 1057.4831.

Step 2: (S) -3- (2-(((8S, 14R) -14- (tert-butoxycarbonyl) -8-((S) -sec-butyl) -1- (9H-fluorene -9-yl) -3,6,9,12-tetraoxo-2-oxa-4,7,10,13-tetraazapentadecane-15-yl) thio) -6-((tert-butoxycarbo Yl) amino) -1H-indol-3-yl) -2-((tert-butoxycarbonyl) amino) propanoic acid (5)

To 4 (3.0 g, 2.83 mmol) compound solution prepared in step 1, dissolved in DCE (200 mL) at rt, Me ₃ SnOH (2.56 g, 14.2 mmol) was added and stirred at 80 for 12 h. TLC analysis confirmed complete consumption of starting material. The reaction mixture was concentrated to remove DCE, dissolved in EtOAc (150 mL) and washed with aqueous KHSO 4. Sat organic layer. Washed with brine, dried over sodium sulfate and concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography using MeOH / MC (1: 4) as eluent to afford the title compound 5 (2.07 g, 1.98 mmol, 70%) as a brown solid.

¹ H NMR (300 MHz, CD3OD) δ 7.80 (d, J = 7.5 Hz, 2H), 7.63 (t, J = 7.4 Hz, 3H), 7.48 (d, J = 8.5 Hz, 1H), 7.39 (t, J = 7.4 Hz, 2H), 7.30 (t, J = 7.2 Hz, 2H), 6.91 (d, J = 8.6 Hz, 1H), 4.45-4.29 (m, 4H), 4.25-4.14 (m, 2H), 4.05-3.96 (m, 1H), 3.87-3.74 (m, 3H), 3.39 (d, J = 4.2 Hz, 1H), 3.22-3.12 (m, 2H), 3.10-2.98 (m, 1H), 2.03- 1.90 (m, 1H), 1.67-1.58 (m, 1H), 1.54 (s, 9H), 1.41 (s, 9H), 1.34 (s, 9H), 1.28-1.20 (m, 1H), 1.00 (d, J = 6.8 Hz, 3H), 0.93 (t, J = 7.3 Hz, 3H);

¹³ C NMR (126 MHz, CD ₃ OD) δ 174.43, 171.49, 170.72, 159.02, 157.49, 145.20, 142.53, 138.82, 135.97, 128.77, 128.18, 126.24, 120.90, 120.23, 118.11, 83.62, 80.45, 68.31, 60.03 56.25, 54.54, 48.27, 47.38, 47.34, 44.80, 43.47, 37.57, 28.81, 28.70, 28.25, 28.17, 27.38, 27.31, 26.10, 15.93, 11.48;

LC / MS mlz 1044.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₅₃ H ₆₉ N ₇ O ₁₃ S [M ⁺ ] 1043.4674.

Step 3: tert-butyl- (3R, 9S, 15S) -15,19-bis ((tert-butoxycarbonyl) amino) -9-((S) -sec-butyl) -5,8,11, 14-tetraoxo-2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,21-hexadecahydro- [1] thia [4,7, 10,13] tetraazacyclooctadecino [18,17-b] indole-3-carboxylate (6)

To 5 (2.0 g, 1.91 mmol) prepared in step 2 dissolved in DMF (100 mL) at rt, DBU (0.57 mL, 3.83 mmol) was added and stirred for 10 minutes. TLC analysis confirmed complete consumption of starting material. The reaction mixture was diluted with DCM (1900 mL) and EDCI HCl (1.83 g, 9.57 mmol), HOBt (1.46 g, 9.57 mmol) was added at rt, and the reaction mixture was stirred at 28 to 24 h. The reaction mixture was concentrated to remove DCM, added with water, then extracted with EtOAc (2 X 150 mL), the organic layer was washed with brine solution (2 X 150 mL), dried over sodium sulfate and dried under reduced pressure. Concentration gave a crude product. The crude product was purified by column chromatography using MeOH / MC (1: 4) as eluent to afford the title compound 6 (810 mg, 1.0 mmol, 53%) as a yellow solid.

¹ H NMR (300 MHz, CD3OD) δ 7.60 (s, 1H), 7.48 (d, J = 8.6 Hz, 1H), 6.92 (dd, J = 8.6, 1.9 Hz, 1H), 4.46-4.30 (m, 2H ), 4.25 (d, J = 5.9 Hz, 1H), 4.15-4.04 (m, 1H), 4.01-3.92 (m, 1H), 3.90-3.78 (m, 2H), 3.48-3.33 (m, 2H), 3.23-3.10 (m, 1H), 3.08-2.94 (m, 1H), 2.08-1.98 (m, 1H), 1.64-1.56 (m, 1H), 1.53 (s, 9H), 1.33 (s, 9H), 1.30 (s, 9 H), 1.16-1.08 (m, 1 H). 0.97 (d, J = 6.8 Hz, 3H), 0.93 (t, J = 7.3 Hz, 3H);

¹³ C NMR (125 MHz, CD ₃ OD) δ 176.19, 174.06, 172.39, 171.62, 170.60, 157.49, 155.52, 139.11, 136.10, 125.26, 125.04, 120.10, 118.42, 113.70, 102.06, 83.57, 80.79, 80.58, 60.26 57.48, 54.79, 54.31, 44.36, 43.53, 38.37, 37.11, 29.49, 28.79, 28.59, 28.07, 25.91, 16.09, 11.87;

LC / MS mlz 803.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₃₈ H ₅₇ N ₇ O ₁₀ S [M ⁺ ] 803.3888, found 803.3888.

Step 4: (3R, 9S, 15S) -15,19-bis ((tert-butoxycarbonyl) amino) -9-((S) -sec-butyl) -5,8,11,14-tetraoxo -2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,21-hexadecahydro- [1] thia [4,7,10,13] Tetraazacyclooctadecino [18,17-b] indole-3-carboxylic acid (6-1)

To 6 (800 mg, 0.99 mmol) compound solution prepared in step 3 dissolved in DCM (100 mL) at 0 ° C., 4.0 M HCl dissolved in dioxane (15 mL) was added and at rt for 12 h. Stirred. The reaction mixture was distilled off to remove the solvent and dried under vacuum. The crude product was obtained, redissolved in THF (100 mL), Boc 2 O (651 mg, 2.98 mmol) was added at 0, followed by 1.0 M NaHCO 3 (10 mL). The reaction mixture was stirred at rt for 12 h. Upon completion of the reaction, THF was removed and the mixture was oxidized with 1.0 M HCl and then extracted with DCM (2 × 150 mL). The collected organic layer was sat. Washed with brine solution, dried over Na ₂ SO _4, and the solvent was removed under reduced pressure to afford the crude product, which was purified by silica gel column chromatography to give the title compound 6-1 (520 mg, 0.695 mmol, 70%). The compound was prepared as a white solid.

¹ H NMR (300 MHz, CD3OD) δ 7.57 (s, 1H), 7.48 (d, J = 8.6 Hz, 1H), 6.94 (d, J = 8.6 Hz, 1H), 4.45-4.32 (m, 2H), 4.23 (d, J = 5.9 Hz, 1H), 4.14-4.09 (m, 1H), 4.02-3.97 (m, 1H), 3.88-3.74 (m, 2H), 3.52-3.35 (m, 2H), 3.21- 3.15 (m, 1H), 3.09-3.01 (m, 1H), 2.08-1.98 (m, 1H), 1.65-1.57 (m, 1H), 1.54 (s, 9H), 1.33 (s, 9H), 1.24- 1.17 (m, 1 H), 0.97 (d, J = 6.2 Hz, 3H), 0.94 (t, J = 7.3 Hz, 3H);

¹³ C NMR (125 MHz, CD ₃ OD) δ 176.81, 176.09, 174.35, 172.71, 171.43, 157.57, 155.56, 138.79, 135.87, 126.56, 125.06, 119.97, 117.93, 113.73, 102.34, 80.86, 80.57, 68.83, 68.83 57.78, 56.02, 49.85, 44.15, 43.87, 39.78, 37.02, 29.49, 28.82, 28.67, 26.06, 15.96, 11.81;

LC / MS mlz 747.9 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₃₄ H ₄₉ N ₇ O ₁₀ S [M ⁺ ] 747.3262.

Step 5: (2R, 3R, 4S) -4-((2S, 4R) -1- (N2-((3R, 9S, 15S) -15,19-bis ((tert-butoxycarbonyl) amino) -9-((S) -sec-butyl) -5,8,11,14-tetraoxo-2,3,4,5,6,7,8,9,10,11,12,13,14, 15,16,21-hexadecahydro- [1] thia [4,7,10,13] tetraazacyclooctadecino [18,17-b] indole-3-carbonyl) -N4-trityl-L -Asparaginyl) -4-hydroxypyrrolidine-2-carboxamido) -5- (tert-butoxy) -3-methyl-5-oxopentan-1,2-diyl diacetate (7 )

Clear solution of 6-1 (500 mg, 0.668 mmol) compound prepared in step 4 and 1-1 (516 mg, 0.668 mmol) compound prepared in Example 1, dissolved in DMF (20 mL) at 0 ° C. To this, HATU (381 mg, 1.0 mmol) was added followed by DIPEA (0.17 mL, 1.0 mmol) dropwise. The reaction mixture was stirred at rt for 12 h. TLC analysis confirmed complete consumption of starting material. Water was added to the reaction mixture and extracted with EtOAc (2 X 50 mL), then the organic layer was washed with brine solution (2 X 50 mL), dried over sodium sulfate and concentrated under reduced pressure to afford the crude product. The crude product was purified via silica gel column chromatography using MeOH / MC (1: 9) as eluent to afford the title compound 7 (650 mg, 0.432 mmol, 65%) as an off-white solid.

¹ H NMR (300 MHz, CD3OD) δ 7.61 (s, 1H), 7.52 (d, J = 8.6 Hz, 1H), 7.33-7.16 (m, 15H), 6.96 (dd, J = 8.6, 1.7 Hz, 1H ), 4.99-4.92 (m, 1H), 4.73 (t, J = 6.4 Hz, 1H), 4.59-4.52 (m, 1H), 4.49-4.42 (m, 1H), 4.42-4.30 (m, 3H), 4.24 (dd, J = 12.4, 2.2 Hz, 1H), 4.18 (d, J = 6.9 Hz, 1H), 4.03-3.95 (m, 1H), 3.93 (d, J = 5.9 Hz, 1H), 3.90 (d , J = 4.0 Hz, 1H), 3.86-3.79 (m, 1H), 3.78-3.68 (m, 1H), 3.60-3.52 (m, 1H), 3.47-3.38 (m, 2H), 3.18-3.07 (m , 1H), 3.05-2.95 (m, 1H), 2.81 (d, J = 6.5 Hz, 2H), 2.26-2.15 (m, 2H), 2.10-2.01 (m, 2H), 1.99 (s, 3H), 1.98-1.94 (m, 1H), 1.91 (s, 3H), 1.66-1.58 (m, 1H), 1.55 (s, 9H), 1.48 (s, 9H), 1.37 (s, 9H), 1.25-1.14 ( m, 2H), 1.00-0.88 (m, 8H), 0.77 (d, J = 7.0 Hz, 1H).

¹³ C NMR (125 MHz, CD ₃ OD) δ 175.61, 174.38, 173.99, 172.43, 172.34, 172.12, 172.02, 171.54, 170.98, 170.76, 157.48, 155.47, 145.71, 145.69, 139.06, 136.04, 130.08, 128.79,128. 125.51, 125.03, 120.24, 118.33, 113.60, 83.21, 83.16, 80.80, 80.57, 74.01, 71.73, 71.58, 70.74, 70.30, 64.63, 64.51, 60.92, 60.60, 60.37, 56.96, 56.29, 55.26, 55.22, 54.81, 50.81, 54.81 49.85, 44.22, 43.75, 39.32, 38.74, 37.27, 36.73, 28.83, 28.68, 28.41, 26.04, 21.02, 20.71, 16.02, 13.49, 11.63;

LC / MS mlz 1503.2 [M + H ⁺ ].

HRMS (EI) m / z calcd for C ₇₆ H ₉₉ N ₁₁ O ₁₉ S [M ⁺ ] 1501.6839.

Step 6: (2S, 3R, 4R) -4,5-Diacetoxy-2-((2S, 4R) -1-(((3R, 9S, 15S) -15,19-diamino-9- ( (S) -sec-butyl) -5,8,11,14-tetraoxo-2,3,4,5,6,7,8,9,10,11,12,13,14,15,16, 21-hexadecahydro- [1] thia [4,7,10,13] tetraazacyclooctadecino [18,17-b] indole-3-carbonyl) -L-asparaginyl) -4-hydro Roxypyrrolidine-2-carboxamido) -3-methylpentanoic acid (8)

At 0 ° C, 2.5 mL of 1.0 N HCl dissolved in HFIP (5.0 mL 4.0 N HCl dissolved in dioxane) in 7 (100 mg, 0.066 mmol) compound solution prepared in step 5 dissolved in DCM (5.0 mL) And 15.0 mL of HFIP were mixed). Stir at rt for 12 h and notice that the reaction mixture immediately changed to a dark yellow color. The progress of the reaction was monitored via LCMS. The reaction mixture was concentrated under reduced pressure to remove the solvent and dried under high vacuum. The resulting solid was washed with nucleic acid, filtered and dried under reduced pressure to give the title compound 8 (65 mg, 0.064 mmol, 98%) as a pale brown solid. It was used for the next step without further purification.

LC / MS mlz 1004.1 [M + H ⁺ ]; 1002.0 [M-H ⁺ ].

Step 7: Compound represented by Formula 9

PyAOP (41 mg, 0.077 mmol) and HOAt (1.8 mg) at 0 ° C. in 8 (65 mg, 0.065 mmol) compound solution prepared in step 6 dissolved in DMF / DCM (1: 1) (10.0 mL). , 0.013 mmol) was added, followed by DIPEA (34 μL, 0.194 mmol) dropwise. The reaction mixture was stirred at rt for 12 h. The progress of the reaction was monitored by LCMS. The reaction mixture was concentrated at 15 under reduced pressure to remove DCM. Water (10 mL) was added and stirred for 2 h. The solid precipitate was filtered off and lyophilized under reduced pressure to remove water and DMF. The crude product formed was purified via HPLC using 0.1% TFA dissolved in CAN and 0.1% TFA dissolved in H 2 O as eluent. The desired fractions obtained from HPLC were lyophilized under reduced pressure to give the title compound 9 (26 mg, 0.026 mmol, 40%) as a white solid.

¹ H NMR (500 MHz, CD3OD) δ 8.88 (dd, J = 7.6, 5.0 Hz, 1H), 8.80 (d, J = 3.0 Hz, 1H), 8.53 (t, J = 8.8 Hz, 2H), 8.45 ( d, J = 3.5 Hz, 1H), 8.42 (t, J = 8.5 Hz, 2H), 8.24 (s, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.46 (s, 1H), 7.31 ( d, J = 2.0 Hz, 1H), 7.04 (dd, J = 8.5, 2.1 Hz, 1H), 5.26 (p, J = 3.9 Hz, 1H), 5.15 (p, J = 6.8 Hz, 1H), 4.82- 4.72 (m, 2H), 4.62-4.52 (m, 3H), 4.43 (dd, J = 18.2, 8.9 Hz, 1H), 4.34 (dd, J = 11.9, 4.0 Hz, 1H), 4.20-4.10 (m, 2H), 4.01 (dd, J = 11.2, 3.3 Hz, 1H), 3.78 (d, J = 10.9 Hz, 1H), 3.74-3.65 (m, 2H), 3.60 (d, J = 18.0 Hz, 1H), 3.54 (dd, J = 17.7, 4.6 Hz, 1H), 3.49-3.43 (m, 1H), 3.28 (dd, J = 14.9, 6.8 Hz, 1H), 3.24-3.18 (m, 1H), 3.12 (dd, J = 16.1, 4.0 Hz, 1H), 3.02 (dd, J = 10.9, 4.0 Hz, 1H), 2.53-2.47 (m, 1H), 2.46-2.41 (m, 1H), 2.13 (td, J = 12.7, 3.6 Hz, 1H), 2.09 (s, 3H), 2.05 (s, 3H), 1.73-1.59 (m, 2H), 1.26-1.18 (m, 1H), 1.11 (d, J = 7.2 Hz, 3H), 0.97-0.89 (m, 6H). ¹ H NMR (500 MHz, D ₂ O) δ 7.71 (d, J = 8.5 Hz, 1H), 7.34 (s, 1H), 7.04 (d, J = 8.8 Hz, 1H), 5.18-5.13 (m, 1H ), 4.87 (t, J = 5.3 Hz, 1H), 4.82-4.74 (m, 2H), 4.69-4.62 (m, 2H), 4.64-4.56 (m, 1H), 4.41-4.32 (m, 2H), 4.16-4.08 (m, 2H), 4.07 (d, J = 3.1 Hz, 1H), 3.99-3.95 (m, 1H), 3.89-3.82 (m, 2H), 3.71 (d, J = 17.6 Hz, 1H) , 3.66-3.57 (m, 2H), 3.46-3.40 (m, 1H), 3.21 (dd, J = 14.1, 6.5 Hz, 1H), 3.14-3.09 (m, 1H), 3.07-3.01 (m, 2H) , 2.59-2.51 (m, 1H), 2.43 (dd, J = 13.8, 7.0 Hz, 1H), 2.15-2.11 (m, 1H), 2.10 (s, 3H), 2.03 (s, 3H), 1.72-1.63 (m, 1H), 1.54-1.44 (m, 1H), 1.23-1.12 (m, 1H), 1.02 (d, J = 7.1 Hz, 3H), 0.85-0.78 (m, 6H).

¹³ C NMR (125 MHz, D2O) δ 175.04, 174.36, 173.84, 173.40, 173.37, 172.47, 172.06, 171.33, 170.69, 170.13, 169.77, 136.26, 127.14, 126.37, 126.21, 120.84, 115.26, 114.08, 115.26, 114.08. 69.89, 63.19, 61.94, 59.42, 56.40, 55.43, 54.53, 52.20, 51.08, 42.57, 41.80, 37.44, 37.15, 35.78, 34.87, 34.48, 28.06, 25.17, 20.56, 20.13, 14.24, 11.82, 9.63;

LC / MS mlz 987.5 [M + H ⁺ ], m / z 984.6 [M-H ⁺ ],

HRMS (EI) m / z calcd for C ₄₃ H ₅₉ N ₁₁ O ₁₄ S [M ⁺ ] 985.3964.

Step 8: Compound represented by Formula 10

From 0 ℃, MeOH (1.0 mL) of the above step at a 9 prepared in 7 (10 mg, 0.01 mmol) compound solution, adding the 0.02MK ₂ CO ₃ was dissolved in MeOH (1.0 mL), and the reaction mixture was dissolved in Stir at rt for 2 h. The progress of the reaction was monitored by LCMS. The reaction mixture was concentrated under reduced pressure at 15 to remove MeOH. The crude reaction mixture was purified via HPLC using 0.1% TFA dissolved in ACN and TFA dissolved in H ₂ O as eluent. The desired fractions obtained from HPLC were lyophilized under reduced pressure to give the title compound 10 (5.3 mg, 0.0058 mmol, 58%) as a white solid.

¹ H NMR (500 MHz, CD3OD) δ 8.96-8.89 (m, 1H), 8.67-8.61 (m, 1H), 8.53 (d, J = 3.9 Hz, 1H), 8.39 (d, J = 9.7 Hz, 1H ), 8.31 (d, J = 7.6 Hz, 1H), 8.14 (d, J = 9.8 Hz, 1H), 8.03 (d, J = 9.2 Hz, 1H), 7.93 (d, J = 8.5 Hz, 1H), 7.32 (d, J = 2.0 Hz, 1H), 7.05 (dd, J = 8.5, 2.0 Hz, 1H), 5.45-5.39 (m, 1H), 4.83 (d, J = 3.3 Hz, 1H), 4.82-4.77 (m, 1H), 4.73 (dd, J = 9.8, 4.3 Hz, 1H), 4.65-4.57 (m, 2H), 4.31 (dd, J = 18.3, 8.2 Hz, 1H), 4.21-4.13 (m, 1H ), 4.00 (dd, J = 11.2, 3.3 Hz, 1H), 3.81 (d, J = 10.7 Hz, 1H), 3.75 (dd, J = 8.4, 3.8 Hz, 1H), 3.68-3.65 (m, 1H) , 3.63 (d, J = 2.6 Hz, 1H), 3.60-3.58 (m, 1H), 3.58-3.55 (m, 1H), 3.51-3.46 (m, 1H), 3.44-3.39 (m, 1H), 3.37 -3.35 (m, 1H), 3.24 (d, J = 6.0 Hz, 1H), 3.22-3.17 (m, 2H), 3.05 (dd, J = 11.1, 4.0 Hz, 1H), 2.62-2.56 (m, 1H ), 2.47 (dd, J = 13.2, 6.9 Hz, 1H), 2.09 (td, J = 12.6, 3.6 Hz, 1H), 1.74-1.68 (m, 1H), 1.67-1.61 (m, 1H), 1.27- 1.20 (m, 1 H), 1.01 (d, J = 7.0 Hz, 3H), 0.96-0.90 (m, 6H);

LC / MS mlz 902.72 [M + H ⁺ ], m / z 900.77 [M-H ⁺ ].

Claims (10)

A compound represented by formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula I]

(In Formula I,
n is an integer from 0 to 2;

R ¹ is —NH ₂ , —NHR, or —NR ₂ ,
Wherein R is C _1-10 straight or branched chain alkyl, or a protecting group; And

R ² , R ³ , R ⁴ and R ⁵ are independently —H, —OH, —NH ₂ , or C _1-10 straight or branched chain alkyl).
The method of claim 1,
n is an integer from 0 to 2;

R ¹ is —NH ₂ , —NHR, or —NR ₂ ,
Wherein R is C _1-5 straight or branched alkyl, or a protecting group; And

R ² , R ³ , R ⁴ and R ⁵ are independently —H, —OH, —NH ₂ , or C _1-5 straight or branched chain alkyl, optical isomers thereof, or pharmaceuticals thereof Acceptable salts.
The method of claim 1,
n is an integer from 0 to 2;

R ¹ is —NH ₂ , —NHR, or —NR ₂ ,
Wherein R is C _1-3 straight or branched alkyl, or a protecting group;

R ² is -NH ₂ , or -OH; And
R ³ , R ⁴ and R ⁵ are independently -H, or -OH, a compound, optical isomer thereof, or a pharmaceutically acceptable salt thereof.
The method according to any one of claims 1 to 3,
The protecting group is a compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that any one selected from the group consisting of:
Acetyl (Ac), Acetoxy (OAc), β-Methoxyethoxymethyl ether (MEM), Dimethoxytrityl, Methoxymethyl ether (MOM) , Methoxytrityl, p-Methoxybenzyl ether (PMB), methylthiomethyl ether, Pivaloyl (Piv), triphenylmethyl, tert -Butyloxycarbonyl (Boc), 9-fluorenylmethyloxycarbonyl (Fmoc), tosyl, Ts and tert-Butyl esters.
The method of claim 1,
The compound represented by the formula (I) is a compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that:

.
As shown in Scheme 1 below,
Reacting the compound represented by the formula (II) with the compound represented by the formula (III) to prepare a compound represented by the formula (IV);
Preparing a compound represented by Formula V from the compound represented by Formula IV prepared in the above step;
Preparing a compound represented by Chemical Formula VI through a cyclization reaction through dehydration of the compound represented by Chemical Formula V prepared in the above step; And
A method for preparing a compound represented by the formula (I) of claim 1, comprising the step of preparing a compound represented by the formula (I) by removing a protecting group from the compound represented by the formula (VI) prepared in the step:
Scheme 1

(In Scheme 1,
L is -NH-, or -NR-;
n is an integer from 0 to 2;
R ¹ is —NH ₂ , —NHR, or —NR ₂ ,
Wherein R is C _1-10 straight or branched chain alkyl, or a protecting group; And
R ² , R ³ , R ⁴ and R ⁵ are independently —H, —OH, —NH ₂ , or C _1-10 straight or branched chain alkyl; And
PG ₁ to PG ₃ are protecting groups;

To
The compound represented by Formula II is as shown in Scheme 2 below,
Reacting the compound represented by the formula (X1) with the compound represented by the formula (X2) to prepare a compound represented by the formula (X3);
Preparing a compound represented by Chemical Formula X4 by converting the -OMe group of the compound represented by Chemical Formula X3 prepared in the above step into -OH;
Preparing a compound represented by Chemical Formula X5 by cyclizing the compound represented by Chemical Formula X4 prepared in the above step; And
Method for preparing a compound represented by the formula (II) by converting the -OtBu group of the compound represented by the formula (X5) prepared in the above step into -OH:
Scheme 2

(In Scheme 2,
L is -NH-, or -NR-;
n is an integer from 0 to 2; And
PG ₁ and PG ₄ are protecting groups.
The method of claim 6,
L in Scheme 1 is -NH-;
R ¹ and R ² are -NH ₂ ;
R ⁴ and R ⁵ are -OH;
PG ₁ is tert-Butyloxycarbonyl (Boc);
PG ₂ is acetoxy (OAc); And
PG ₃ is triphenylmethyl (triphenylmethyl) characterized in that the manufacturing method.
The method of claim 6,
Compound represented by Formula III is shown in Scheme 3 below,
Reacting the compound represented by the formula (Y1) with the compound represented by the formula (Y2) to prepare a compound represented by the formula (Y3); And
Process for preparing a compound represented by the formula (III) by removing the PG ₄ protecting group of the compound represented by the formula Y3 prepared in the step:
Scheme 3

(In Scheme 3,
R ³ is —H, —OH, —NH ₂ , or C _1-10 straight or branched chain alkyl; And
PG ₂ to PG ₄ are protecting groups).
A compound represented by the following formula α, an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula α]

(In the above formula α,
n and L are as defined in Scheme 1 of claim 6; And
A ¹ and A ² are protecting groups.
A pharmaceutical composition for preventing or treating cancer, comprising the compound represented by the formula (I) of claim 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.