KR100225416B1 - Peptidomimetic ras transformed cell growth inhibitors, process for preparation thereof, and composition comprising thereof - Google Patents

Abstract

The present invention relates to novel analogous peptide derivatives represented by general formula (I), nontoxic salts thereof, methods for their preparation, and Ras mutant cell growth inhibitory compositions containing them as active ingredients.

Wherein R ₁ is hydrogen, straight or branched C ₁ -C ₃ alkyl, cyano substituted straight or branched C ₁ -C ₃ alkyl, or acetyl, and A is -CH ₂ -or -CO- . Also, X is a naphthalene, Chi-2-yl, C ₁ - C ₃ alkoxy, phenyl, or C substituted with ₁ - C ₃ alkoxy substituted with C ₁ - C ₃ alkyl and, Y is naphthalene, quinoline, thiophene value -2-yl, thiopen-2-yl-methyl, phenoxymethyl, benzyloxymethyl, ethoxycarbonylethyl, phenyl with or without substituents, phenylmethyl with or without substituents, with or without substituents Is straight or branched C ₁ -C ₅ alkyl or alkenyl, or C ₃ -C ₅ cycloalkyl.

Description

Similar peptide derivatives, preparation methods thereof, and ras transformed cell growth inhibitors containing them as active ingredients

The present invention relates to novel analogous peptide derivatives and nontoxic salts thereof, and more particularly to novel analogous peptide derivatives represented by the following general formula (I) and their nontoxic salts, preparation methods thereof, and their active ingredients It relates to a Ras (mutation cell growth inhibitory composition) containing.

Wherein R ₁ is hydrogen, straight or branched C ₁ -C ₃ alkyl, cyano substituted straight or branched C ₁ -C ₃ alkyl, or acetyl, and A is -CH ₂ -or -CO- . Also, X is a naphthalene, Chi-2-yl, C ₁ -C ₃ alkoxy, and phenyl, or C ₁ -C ₃ alkyl substituted with a C ₁ -C ₃ alkoxy substituted by, Y is naphthalene, quinoline, thiophene value -2-yl, thiopen-2-yl-methyl, phenoxymethyl, benzyloxymethyl, ethoxycarbonylethyl, phenyl with or without substituents, phenylmethyl with or without substituents, with or without substituents Is straight or branched C ₁ -C ₅ alkyl or alkenyl, or C ₃ -C ₅ cycloalkyl.

Mammalian Ras protein is a protein synthesized by the Ras gene and is called two prenyltransferases, namely Farnesyl protein transferase (FPFP). Or by means of Geranylgeranyl protein transferase (hereinafter referred to as "GGPT"), farnesylation or Geranylgeranylation to the cell wall, which is linked to GDP It is either in an activated state or in an activated state with GTP bound. When the Ras protein is activated, it transmits signals outside the cell into the nucleus by activating the lower level signaling system step by step. This information activates transcription factors (myc, jun, fos, etc.) to grow cells or This series of information transmission systems is called the Ras signaling system (M. Barbacid, Annu. Rev. Biochem., 56, 779, 1987, P J. Casey et al., Natl. Acad. Sci). USA 86, 8323, 1989).

On the other hand, when mutagenic Ras proteins are generated by mutations of the Ras gene (e.g., H-Ras, N-Ras, K-RasA, K-RasB, etc.), that is, they bind to GTP and remain active. It causes cancer of the cells. Among many human cancers, especially in patients with colorectal cancer and pancreatic cancer, these mutant Ras genes are found to be about 50% and 90%, respectively, and Ras genes mutated at high frequency in lung cancer (50%) or thyroid cancer (30%). It has been confirmed (S. Rodenhuis, Semin. Cancer Biol. 3, 241, 1992).

Therefore, many studies have been conducted to develop mutant cell inhibitors due to mutant Ras proteins. In particular, a lot of research is being conducted on FPT inhibitors that can block the transfer of mutant Ras protein into the cell wall.

For example, Cys-Val-Phe-Met, which has a structure similar to the carboxylic acid residue (Cys-A1-A2-Met) of the las protein, has been described as an inhibitor of FPT (JL Goldstein et al., J Biol. Chem., 266. 15575, 1991; AM Garcia et al., J. Biol. Chem., 268, 18415. 1993; SL Graham et al., J. Med. Chem., 37, 725, 1994). .

In addition, a number of derivatives are under development based on the Cys-lle-Phe-Met as a basic structure, and there are reports that a compound in which the Phe-Met moiety is replaced with an aromatic alkylamine selectively inhibits FPT compared to GGPT. (SJ Desolms et al., J. Med. Chem., 38, 3967, 1995), that carbosylamide compounds that bind aminomethylnaphthalene to cysteine and trans-3 (S) -ethylproline show inhibition against FPT As disclosed (WO9606609, 1996), similar peptide compounds obtained by converting cysteine to imidazole ethyl groups have also been reported to have an FPT inhibitory effect (JH Hunt et al., J. Med. Chem., 39, 353, 1996; WO9610035, 1996; WO9610034, 1996; WO9609836, 1996). In addition, Merck reports that compounds converting cysteine from Cys-lle-Phe-Met to p-nitrobenzylimidazole acetate and phenylalanine to (N-naphthylmethyl) glycine have inhibitory effects on FPT ( WO9610034, 1996).

However, it has recently been reported that such conventionally developed FPT inhibitors do not effectively inhibit farnesylation in K-Ras mutant cells found in most human cancers. In other words, K-RasB, which is the most frequently found in human cancer cells, maintains the activity of Ras protein by prenylating by GGPT (particularly GGPT-1) when FPT inhibitor inhibits farnesylation of Ras protein. (GL James et al., J. Biol. Chem. 270, 6221, 1995).

In addition, the conventional FPT inhibitors are being developed as a peptide compound having two or more amide bonds and a carboxylic acid, has a low cell permeability and can be easily hydrolyzed by peptidase.

Therefore, the present inventors not only inhibit the growth of the Ras-mutant cells themselves, which are completely different from the conventional Ras protein production inhibitors having FPT or GGPT inhibitory mechanisms, but also by a peptidase which is a problem with the conventional peptide compounds. As a result of researches to develop a novel las protein growth inhibitor which can reduce the degradation effect, the present inventors have found that similar peptide derivatives of the general formula (I) can effectively inhibit the growth of Ras mutant cells. .

Hereinafter, the present invention will be described in detail.

It is an object of the present invention to provide novel analogous peptide derivatives having a las mutant cell growth inhibitory effect and nontoxic salts thereof.

It is also an object of the present invention to provide a method for preparing the analogous peptide derivatives and nontoxic salts thereof.

It is also an object of the present invention to provide a Ras mutant cell growth inhibitory composition containing the analogous peptide derivatives and non-toxic salts thereof as an active ingredient.

Hereinafter, the present invention will be described in more detail.

The present invention includes novel analogous peptide derivatives represented by the following general formula (I) and nontoxic salts thereof.

Wherein R ₁ is hydrogen, straight or branched C ₁ -C ₃ alkyl, cyano substituted straight or branched C ₁ -C ₃ alkyl, or acetyl, and A is —CH ₂ — or —CO— . Also, X is a naphthalene, Chi-2-yl, C ₁ -C ₃ alkoxy, and phenyl, or C ₁ -C ₃ alkyl substituted with a C ₁ -C ₃ alkoxy substituted by, Y is naphthalene, quinoline, thiophene value -2-yl, thiopen-2-yl-methyl, phenoxymethyl, benzyloxymethyl, ethoxycarbonylethyl, phenyl with or without substituents, phenylmethyl with or without substituents, with or without substituents Straight chain or branched C ₁ -C ₅ alkyl or alkenyl, or C ₃ -C ₅ cycloalkyl.

The analogous peptide derivatives according to the invention may be in the form of pharmaceutically acceptable salts, which may be in the form of non-toxic salts commonly used in the field of anticancer agents, for example salts formed from non-toxic inorganic or organic acids. . Such conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and the like and acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, pamoic acid, maleic acid, hydroxymale Salts prepared from organic acids such as acids, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxy-benzoic acid, fumaric acid, toluenesulfonic acid, methanedisulfonic acid, ethanedisulfonic acid, oxalic acid, trifluoroacetic acid, and the like. do.

The analogous peptide derivative according to the present invention represented by the general formula (I) may be in the form of various stereoisomers by containing the absent carbon as represented by the following general formula, and may also be in a mixed form thereof. Therefore, unless otherwise indicated, the present invention is to be understood to encompass all of them.

Examples of the more preferable compound among the compounds according to the present invention are as follows.

The present invention includes methods for producing analogous peptide derivatives represented by the general formula (1) and nontoxic salts thereof. The preparation method of these derivatives is represented as follows.

Scheme 1

Scheme 2

In the above scheme, A, R ₁ , X and Y are the same as defined above, Boc is a common amino protecting group such as t-butoxycarbonyl and Pr is a sulfite which can be removed simultaneously with the amino protecting group (Boc). As a protecting group of a drill machine, it is triphenylmethyl, St-butyl, etc., for example.

The reaction scheme is described for each step as follows.

[Reaction 1]

Reductive alkylation reaction

Isoleucine aldehyde derivatives protected with t-butoxycarbonyl (Boc) of the general formula (II) as a starting material are known methods (Org. Syn. Vol. 67, 1988, 69 and Synthesis, 1983, 676). The compound of formula (III) may be prepared by performing a reductive alkylation reaction by adding X-methylamine in the presence of a known reducing agent such as sodium cyanoborohydride. At this time, the reaction can be promoted by adding KOAc and 3A molecular sieve.

[Reaction 2]

Acylation or alkylation reactions

Compounds of formula (IV) can be prepared by reacting a compound of formula (III) with a suitable fusion agent or by reacting Y-carbonyl halide in the presence of a suitable base. As a fusion agent which can be used together with Y-carboxylic acid, hydroxybenzotriazole, dialkyl carbodiimide, or a mixed solution thereof can be used. Dimethylformamide, dichloromethane, or a mixed solution thereof may be used as the solvent.

[Reaction 3]

Removal reaction of amino protecting group

The compound of formula (IV) having an amino protecting group is dissolved in a suitable organic solvent such as dichloromethane and then reacted with a deprotecting agent such as trifluoroacetic acid to remove the protecting group.

[Reaction 4]

Acylation or Reductive Alkylation

[Reaction 4-1]

Acylation (if A is -CO-)

Fusion of hydroxybenzotriazole and dialkylcarbodiimide to a cysteine (e.g., Nt-butoxycarbonyl-S-triphenylmethyl-L-cysteine) having an appropriate protecting group attached to the compound of formula (V) The acylation reaction is carried out in the presence of a suitable base such as N-methylmorpholine to prepare a compound of formula (VI) wherein A is -CO-.

[Reaction 4-2]

Reductive alkylation (if A is -CH _2- )

A cysteine aldehyde (e.g., Nt-butoxycarbonyl-S-triphenylmethyl-L-cysteine aldehyde) having an appropriate protecting group attached to the compound of formula (V) is reacted with a reducing agent such as sodium cyanoborohydride. Reduction by addition may yield a compound of formula (VI) wherein A is -CH _2- . At this time, the reaction can be promoted by adding KOAc or 3A molecular sieve.

[Reaction 5]

Deprotector Reaction

To the compound of formula (VI) protected with an amino protecting group and a sulfhydryl group protecting group, an amino protecting group and a sulfhydryl protecting group are simultaneously removed by adding a carbonium ion remover such as triethylsilane under acidic conditions such as trifluoroacetic acid. To prepare a compound of formula (I) wherein R ₁ is hydrogen.

[Reaction 6]

Alkylation and Deprotector Reactions

Compound of formula (VII) is prepared by reacting a compound of formula (VI) protected with an amino protecting group and a sulfhydryl group with a R ₁ halide in the presence of a suitable base such as K ₂ CO ₃ to carry out an alkylation reaction. The deprotection group reaction may then be carried out in the same manner as in reaction 5 to prepare a compound of formula (I) in which R ₁ is not hydrogen.

Pharmaceutically acceptable non-toxic salts of the compounds according to the invention can be prepared by conventional methods from compounds of the invention containing basic moieties. In general, salts may be prepared by reacting an organic base in a stoichiometric amount or in excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents.

The present invention includes a Ras mutant cell growth inhibitory composition containing a similar peptide derivative represented by the general formula (I) as an active ingredient and a pharmaceutically acceptable carrier. The composition according to the present invention may include excipients such as lactose, corn starch, lubricants such as magnesium stearate, emulsifiers, suspending agents, buffers, isotonic agents and the like which are well known and can be used. It may include the agent.

The composition according to the invention can be administered orally or parenterally, including intravenous, intraperitoneal, subcutaneous, rectal and topical administration. That is, the composition according to the present invention may be administered in the form of a tablet or capsule, or as an aqueous solvent or suspension. In the case of oral tablets, carriers commonly used include lactose and corn starch, and lubricants such as magnesium stearate can usually be added. Useful diluents in the case of capsular forms may include lactose and dry corn powder. If an aqueous suspension is required for oral use, the active ingredient may include emulsifiers and suspensions. If desired, certain sweetening and / or flavoring agents may be added. For intramuscular, intraperitoneal, subcutaneous and intravenous administration, a sterile solution of the active ingredient is usually prepared and may be included as a buffer to suitably adjust the pH of the solution. For intravenous administration, the total concentration of the solute is It may include isotonic agents that can be adjusted to impart isotonicity to the formulation. In addition, the composition according to the present invention may be in the form of an aqueous solution containing a pharmaceutically acceptable carrier such as saline having a pH of 7.4, and may be locally introduced into the patient's intramuscular blood flow in the form of a solution. have.

The compound according to the present invention can be administered to cancer patients such as colon cancer, rectal cancer, pancreatic cancer, or myeloid leukemia by effectively inhibiting the growth of las mutant cells, and the dosage is usually the age, weight and response of each patient, and the patient. It may be administered at a dose that can be varied according to the symptoms of, for example, about 0.1 mg / kg to about 20 mg / kg per day, preferably 0.5 mg / kg to about 10 mg / kg per day.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, this does not limit the scope of the invention.

Reference Example 1

Preparation of N-t-butoxycarbonyl-L-isoleucine aldehyde

[Step 1]

N-t-butoxycarbonyl-L-isoleucine-N-methyl, N-methoxyamide

To a suspension of N, O-dimethylhydroxyamine (6.1 g, 62.5 mmol) in dichloromethane (60 mL) was added dropwise N-methylmorpholine (7.6 mL, 69.1 mmol) at -10 < 0 > C, followed by -10 Store at ° C. In another vessel, Nt-butoxycarbonyl-L-isorucin (15 g, 62.4 mmol) was dissolved in dichloromethane (300 mL) and the temperature was cooled to -20 ° C, followed by N-methyl morpholine (7.6 mL, 69.1 mmol) and isobutylchloroformate (8.1 mL, 62.4 mmol) were added dropwise. The temperature was kept at -20 ° C during the dropwise addition. After 5 minutes, N, O-dimethylhydroxyamine solution was added dropwise to the mixture, followed by stirring overnight at room temperature. Water (100 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (100 mL × 3). The organic layer was washed with water (100 mL × 2) and brine (100 mL) solution, then excess water was removed with MgSO ₄ and concentrated. The concentrate was purified by column chromatography with EtOAc / n-Hex (1/4) solution and then concentrated to give the title compound (15.1 g, 89%) as a colorless oil.

[Step 2]

N-t-butoxycarbonyl-L-isoleucine aldehyde

2.4 g (54.7 mmol) of lithium aluminum hydride are suspended in 300 mL of anhydrous tetrahydrofuran and cooled to −45 ° C., and then dissolved in 30 mL of anhydrous tetrahydrofuran in Nt-butoxycarbonyl-L-isoleucine A small amount of N-methyl, N-methoxyamide (15 g, 54.7 mmol) solution was added to bring the temperature between -45 ° C and -35 ° C. After completion of the dropwise addition, the cooling bath was removed to bring the temperature to 5 ° C., and then the reaction solution was cooled to −35 ° C., and KHSO ₄ (12.7 g, 35 mL) aqueous solution was slowly added dropwise. After completion of the dropwise addition, the cooling bath was removed to raise the temperature to room temperature, and after 1 hour, celite was added to collect the aluminum salt. The reaction solution was filtered, and the aluminum salt was washed with EtOAc. The collected organic layers were concentrated under reduced pressure, EtOAc was added, washed with water (100 mL), and the excess water was removed with anhydrous Na ₂ SO ₄ , and concentrated to give Nt-butoxycarbonyl-L-isoleucine aldehyde (8.2 g, 69%) was obtained as a colorless oil. At this time, the temperature was not to exceed 30 ℃, was used in the next reaction without purification.

Reference Example 2

Preparation of N-t-butoxycarbonyl-S-triphenylmethyl cysteine aldehyde

The title compound was prepared in the same manner as in Reference Example 1 using N-t-butoxycarbonyl-S-triphenylmethyl cysteine.

Example 1

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S)- Methylpentylamine

[Step 1]

1- (thiophen-2-yl-methylamino) -2 (S)-(t-butoxycarbonyl) amino-3 (S) -methylpentylamine

2-thiophenmethylamine (1.3 g, 11.6 mmol) was dissolved in methanol (15 mL) and Nt-butoxycarbonylisoleucine aldehyde (2.5 g, 11.6 mmol) prepared in Reference Example 1, KOAc (1.14 g, 11.6 mmol) and 3A molecular sieves (0.5 g) were added. 11.6 ml of sodium cyanoborohydride solution (1M) was added dropwise thereto, followed by stirring overnight at room temperature. The precipitate of the reaction mixture was filtered off and then 100 mL of water was added and extracted with EtOAc (100 mL). The organic layer was concentrated with anhydrous MgSO ₄ to remove excess water. The concentrate was column chromatographed with EtOAc / n-Hex (1/2) solution and concentrated to afford the desired compound (1.62 g, 45%) as a yellow oil.

[Step 2]

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) amino-2 (S)-(t-butoxycarbonyl) amino-3 (S) -methylpentylamine

The compound prepared in step 1 (0.5 g, 1.6 mmol) was dissolved in a solution of dimethylformumamide / dichloromethane (3 mL / 7 mL), 1-naphthoic acid (0.33 g, 1.92 mmol), EDCl (0.37 g, 1.93 mmol) ) And HOBt (0.26 g, 1.92 mmol) were added and stirred at room temperature for 5 hours. Water (100 mL) was added to the reaction mixture, which was then extracted with EtOAc (100 mL × 3). The organic layer was washed with water (100 mL × 2) and saturated brine (100 mL) solution, then excess water was removed with anhydrous MgSO ₄ and concentrated. The concentrate was column chromatographed with EtOAc / n-Hex (1/7) solution and concentrated to give the desired compound (0.58 g, 78%) as a yellow foam.

[Step 3]

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S) -amino-3 (S) -methylpentylamine

The compound prepared in step 2 (0.5 g, 1.07 mmol) was dissolved in dichloromethane (4.0 mL), and trifluoroacetyl acid (1.0 mL 0 was added and stirred for 4 hours. The reaction mixture was concentrated to give the target compound (0.52 g, 100%) was obtained as a yellow oily phase.

¹ H NMR (CDCl ₃ ) δ 0.60-1.20 (m, 8H), 1.60 (M, 1H), 3.40 (m, 2H), 4.50 (m, 3H), 6.80-8.40 (m, 10H), 12.20 ( brs, 3H)

[Step 4]

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[2 (R)-(t-butoxycarbonyl) amino-3-triphenylmethylthiopropyl ] Amino-3 (S) -methylpentylamine

The compound prepared in step 3 (0.52 g, 1.07 mmol) was dissolved in methanol (5 mL), and Nt-butoxycarbonyl-S-tritylcysteine aldehyde (0.8 g, 1.79 mmol) and KOAc (Prepared in Reference Example 2) were used. 0.28 g, 2.85 mmol) and 3A molecular sieve (1.0 g) were added. 1.2 mL of sodium cyanoborohydride solution (1M) was added dropwise to the reaction solution, followed by stirring overnight at room temperature. After the precipitate of the reaction mixture was filtered off, 100 ml of water was added to the filtrate and extracted with EtOAc (100 mL) to remove excess water with anhydrous MgSO ₄ and concentrated. The concentrate was column chromatographed with EtOAc / n-Hex / EtOH (20/100/1) solution and concentrated to afford the desired compound (0.22 g, 26%) as a clear oil.

[Step 5]

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-(2 (R) -amino-3-mercaptopropyl) amino-3 (S) -methylpentyl Amine

The compound prepared in step 4 (0.22 g, 0.28 mmol) was dissolved in a solution of dichloromethane (3.0 mL), trifluoroacetyl acid (0.9 mL) and triethylsilane (0.4 mL, 2.5 mmol) and stirred at room temperature for 17 hours. It was. The reaction mixture was concentrated, washed with hexane (100 mL × 2) and lyophilized to afford the desired compound (180 mg, 94%) as a trifluoroacetyl acid salt.

¹ H NMR (DMSO-d6 + TFA) δ0.80-1.50 (m, 8H), 2.00 (m, 1H), 2.80-3.80 (m, 8H), 4.70 (brs, 2H), 6.80-8.20 (m, 10H )

Analytical HPLC (gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA in Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-4 min (100 / 0-0 / 100), Rt = 18.44 min.

EXAMPLE 2-5

The following analogous peptide derivatives were prepared by reacting the corresponding compounds as starting materials in the same manner as in Example 1.

Example 2

N- (3-Quinolinecarbonyl) -N- (thiophen-2-yl-methyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) Methylpentylamine

¹ H NMR (DMOS-d6 + TFA) δ 0.80-1.50 (m, 8H), 2.00 (m, 1H), 2.60-3.80 (m, 8H), 4.90 (brs, 2H), 6.90-9.50 (m, 9H);

analytical HPLC (gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA in water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), Rt = 16.45 min

Example 3

N- (1-naphthoyl) -N- (4-methoxybenzyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (DMSO-d6 + TFA) δ 6.0-8.0 (m, 11H), 4.35 (q, 2H), 3.1-4.8 (m, 9H), 2.76 (s, 2H), 0.8-1.3 (m, 9H );

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA in Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), Rt = 17.80 min at 220.

Example 4

N- (1-naphthoyl) -N- (2-methoxyethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl 3 + TFA) δ 7.35-8.20 (m, 7H), 3.30-3.62 (m, 5H), 3.62-4.80 (m, 6H), 3.00-3.30 (m, 4H), 0.80-1.60 (m , 9H);

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile in Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100/0), 35 -45 min (100 / 0-0 / 100), Rt = 17.81 min., 18.24 min.at 220 nm.

Example 5

N- (1-naphthoyl) -N- (3-ethoxypropyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl3 + TFA) δ 7.35-8.20 (m, 7H), 4.20-4.40 (m, 1H), 3.60-4.10 (m, 2H), 3.20-3.60 (m, 6H), 2.10-2.90 (m , 2H), 1.20-2.10 (m, 9H), 0.80-1.25 (m, 9H);

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile in Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100/0), 35 -45 min (100 / 0-0 / 100), Rt = 18.82 min., 19.07 min.at 220 nm.

Example 6

N- (thiophen-2-yl-carbonyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -Methylpentylamine tripuloacetic acid salt

[Step 1]

N- (1-naphthalenemethyl) -2 (S)-(t-butoxycarbonylamino) -3 (S) -methylpentylamine

1-aminomethylnaphthalene (3.00 g, 19.1 mmol) was used instead of 2-thiophenmethylamine, and the reaction was carried out in the same manner as in Step 1 of Example 1, where N- (1-naphthalenemethyl) -2 (S)-( t-butoxycarbonylamino) -3 (S) -methylpentanamine was obtained as a yellow oil phase (1.86 g, 28%).

TLC, RF = 0.20 (EtOAc / n-Hex = 1/2)

1 H NMR (CDCl 3) δ 7.40-8.25 (m, 7H), 4.65 (d, 1H), 4.15-4.40 (m, 2H), 3.60-3.80 (m, 1H), 2.70-2.90 (m, 2H), 1.40-1.70 (m, 13H), 0.80-1.00 (m, 6H)

[Step 2]

N- (thiophen-2yl-carbonyl) -N- (1-naphthalenemethyl) -2 (S)-(t-butoxycarbonylamino) -3 (S) -methylpentylamine

The compound prepared in step 1 (0.92 g, 2.60 mmol) was dissolved in 10 ml of anhydrous dimethylformromamide, 2-thiophencarboxylic acid (1.0 g, 7.80 mmol) and hydroxybenzotriazole (1.05 g, 7.80 mmol) And diisopropylcarbodiimide (1.22 ml, 7.80 mmol) was added. The reaction mixture was stirred overnight at room temperature, then water (100 mL) was added and extracted with ethyl acetate (100 ml × 3). The extracted organic layer was washed with water (100 mL × 2) and brine (100 mL), and excess water was removed with MgSO ₄ and concentrated. The concentrated mixture was column chromatographed with EtOAc / n-Hex (1/3) solution and then concentrated to give the desired product in a clear oily phase (0.59 g, 49%).

TLC Rf = 0.31 (EtOAc / n-Hex = 1/3)

1 H NMR (CDCl 3) δ7.9 (m, 3H), 7.4-7.6 (m, 5H), 7.15 (d, 1H), 6.83 (s, 1H), 5.0-5.6 (dd, 2H), 3.9-4.4 ( m, 2H), 2.95 (d, 1H), 1.45 (s, 9H), 0.8-1.2 (m, 9H)

[Step 3]

N- (thiophen-2yl-carbonyl) -N- (1-naphthalenemethyl) -2 (S)-[2 (R)-(t-butoxycarbonyl) amino-3-triphenylmethylthiopropyl ] Amino-3 (S) -methylpentylamine

The compound prepared in step 2 (0.59 g, 1.26 mmol) was dissolved in 10 mL of 50% trifluoroacetyl acid solution and stirred at room temperature for 2 hours. After the reaction mixture was concentrated, triethylamine was added dropwise until the pH reached 8-9. This mixture was dissolved in methanol (10 mL) and Nt-butoxycarbonyl-S-triphenylmethyl-L-cysteine aldehyde (0.68 g, 1.52 mmol) and KOAc (0.15 g, 1.52 mmol) and 3A prepared in Reference Example 2 Molecular sieve (0.5 g) was added. 1.52 mL of sodium cyanoborohydride solution (1M) was added dropwise thereto, followed by stirring overnight at room temperature. The precipitate of the reaction mixture was filtered and 100 mL of water was added, extracted with ethyl acetate (100 mL), and excess water was removed with MgSO ₄ and concentrated. The concentrate was column chromatographed with EtOAc / n-Hex (1/3) solution and concentrated to afford the desired compound as a white foam (350 mg, 35%).

TLC Rf = 0.20 (EtOAc / n-Hex = 1/3)

1 H NMR (CDCl 3) δ 6.8-8.0 (m, 25H), 5.1-5.5 (m, 2H), 4.8 (d, 1H), 4.4 (dd, 1H), 3.3-3.7 (m, 2H), 2.3- 2.9 (m, 5H), 1.44 (s, 9H), 0.6-1.4 (m, 9H)

[Step 4]

N- (thiophen-2yl-carbonyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S)- Methylpentylamine trifluorouroacetic acid salt

The compound (140 mg, 0.18 mmol) prepared in step 3 was dissolved in a solution of trifuluroacetyl acid (9 mL) and triethylsilane (1 mL) and stirred at room temperature for 2 hours. The reaction mixture was concentrated, washed with n-hexane (100 mL × 2) and power dried to give the title compound as the trifluuroacetylate (91 mg, 74%).

HPLC (gradient, A; 0.1% TFA-Water, B; 0.1% TFA-70% Acetonitrile, 1-21 min (B; 0 → 100%), 21-35 min (100%), 35-55 min (100 → 0%), retention time = 18.38 min at 220 nm

1 H NMR (CDCl 3) δ 6.8-8.0 (m, 10H), 5.2-5.6 (q, 2H), 3.9-4.4 (m, 2H), 2.8-3.8 (m, 6H), 0.7-2.0 (m, 9H)

[Examples 7-13]

The following analogous peptide derivatives were prepared by reacting the corresponding compounds as starting materials in the same manner as in Example 6.

Example 7

N- (3-chlorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.2-8.0 (m, 11H), 5.05 (s, 2H), 4.0-4.3 (m, 3H), 2.9-3.6 (m, 5H), 0.8-1.0 (m, 9H )

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.39 min at 220 nm.

Example 8

N- (3,4-dichlorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentyl Amine

¹ H NMR (CDCl ₃ ) δ 7.2-8.0 (m, 10H), 5.1 (s, 2H), 4.0-4.5 (m, 3H), 2.9-3.6 (m, 6H), 0.8-1.1 (m, 9H )

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 20.55 min at 220 nm.

Example 9

N- (4-methoxybenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 10H), 5.2-5.6 (dd, 2H), 4.6-4.8 (m, 1H), 4.4-4.6 (m, 1H), 3.8-4.1 (m , 2H), 2.6-3.5 (m, 8H), 0.6-2.0 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.05 min, 19.46 min at 220 nm.

Example 10

N- (thiophen-2-yl-methylcarbonyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S ) -Methylpentylamine

¹ H NMR (CDCl ₃ ) δ 6.8-8.0 (m, 10H), 5.2 (d, 2H), 3.0-4.3 (m, 8H), 2.86 (bs, 2H), 0.8-1.3 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 18.36 min at 220 nm.

Example 11

N- (2-methylbenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.20-8.40 (m, 11H), 4.70-5.20 (m, 2H), 3.75-4.30 (m, 2H), 2.50-3.75 (m, 6H), 2.40 (s, 3H ), 1.10-2.00 (m, 3H), 0.40-1.00 (m, 6H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 17.97 min., 18.74 min.at 220 nm.

Example 12

N- (4-methylbenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.20-8.20 (m, 11H), 4.90-5.30 (m, 2H), 3.75-4.40 (m, 2H), 2.70-3.70 (m, 6H), 2.40 (S, 3H ), 1.20-2.00 (m, 3H), 0.40-1.00 (m, 6H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 18.26 min.at 220 nm.

Example 13

N- (phenylacetyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.8-7.4 (m, 9H), 2.8-4.7 (m, 15H), 0.7-1.4 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 15.98 min at 220 nm.

Example 14

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S) -methylpentylamine

[Step 1]

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[N- (2 (R)-(t-butoxycarbonylamino) -3-triphenyl Thiopropyl)]-N-methylamino] -3 (S) -methylpentylamine

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[(2 (R)-(t-butoxycarbonyl) prepared in step 4 of Example 1 Amino) -3-triphenylthiopropylamino] -3 (S) -methylpentylamine (0.21 g, 0.26 mmol) was dissolved in 10 mL of dimethylformamide and K ₂ CO ₃ (1 g) was added and stirred at room temperature. Water (100 mL) was added to the reaction mixture, followed by extraction with EtOAc (100 mL) The organic layer was washed with water (100 mL × 2) and brine (100 mL) solution, then excess water was removed with MgSO ₄ and concentrated. Water was column chromatographed with EtOAc / n-hexane (1/3) solution and concentrated to give the title compound as a white foam (100 mg, 47%).

TLC Rf = 0.41 (EtOAc / n-Hex = 1/3)

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 25H), 5.2-5.8 (m, 2H), 4.85 (d, 1H), 3.6-3.8 (m, 2H), 3.2-3.5 (m, 1H ), 1.9-3.0 (m, 8H), 1.43 (d, 9H), 0.6-1.4 (m, 9H)

[Step 2]

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S) -methylpentylamine

The compound (80 mg, 0.1 mmol) prepared in step 1 was dissolved in a solution of trifluoroacetyl acid (4.5 mL) and triethylsilane (0.5 mL), and stirred at room temperature for 2 hours. The reaction mixture was concentrated, washed with n-Hex (100 mL × 2) and lyophilized to afford the title compound as trifluoroacetyl acid salt (35.8 mg, 52%).

HPLC (gradient, A; 0.1% TFA-Water, B; 0.1% TFA-70% Acetonitrile, 1-21 min (B; 0 → 100%), 21-35 min (B; 100%), 35-55 ( B; 100 → 0%)), retention time = 19.05 min, 19.46 min at 220 nm;

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 10H), 5.2-5.6 (dd, 2H), 4.6-4.8 (m, 1H), 4.4-4.6 (m, 1H), 3.8-4.1 (m , 2H), 2.6-3.5 (m, 8H), 0.6-2.0 (m, 9H);

[Examples 15-24]

The following analogous peptide derivatives were prepared by reacting the corresponding compounds as starting materials in the same manner as in Example 14.

Example 15

N- (3,4-dichlorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S ) -Methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.2-8.0 (m, 10H), 5.1 (s, 2H), 4.0-4.6 (m, 3H), 2.7-3.3 (m, 8H), 0.8-1.35 (m, 9H )

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 21.69 min, 22.06 min at 220 nm.

Example 16

N- (2,4-difluorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 6.8-8.0 (m, 10H), 5.0 (dd, 2H), 4.55 (dd, 1H), 3.8-4.1 (m, 2H), 2.8-3.2 (m, 5H), 2.7 (s, 3H), 0.8-1.36 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 20.05 min at 220 nm.

Example 17

N- (thiophen-2-yl-methylcarbonyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 10H), 5.2-5.6 (dd, 2H), 4.6-4.8 (m, 1H), 4.4-4.6 (m, 1H), 3.8-4.1 (m , 2H), 2.6-3.5 (m, 8H), 0.6-2.0 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.05 min, 19.46 min at 220 nm.

Example 18

N- (4-methoxybenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S) Methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 11H), 5.0-5.2 (dd, 2H), 4.6 (m, 1H), 4.3 (m, 1H), 3.8-4.1 (m, 5H), 2.6-3.3 (m, 8H), 0.6-2.0 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.83 min at 220 nm.

Example 19

N- (2-thiophen-2-yl-methylcarbonyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methyl Amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 6.8-8.0 (m, 10H), 5.2 (d, 2H), 3.0-4.3 (m, 8H), 2.86 (bs, 2H), 0.8-1.3 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.22 min at 220 nm.

Example 20

N- (2-thiophen-2-yl-methylcarbonyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-propyl Amino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 10H), 5.18 (dd, 2H), 2.8-4.2 (m, 12H), 0.6-1.4 (m, 14H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 18.52 min at 220 nm.

Example 21

N- (4-methoxybenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-cyanomethylamino] -3 S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.76-7.9 (m, 11H), 5.03 (dd, 2H), 4.0-4.2 (m, 2H), 3.7-3.9 (m, 5H), 2.7-3.6 (m, 8H ), 0.6-1.4 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 17.93 min at 220 nm.

Example 22

N- (2-methylbenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S)- Methylpentylamine

¹ H NMR (CDCl ₃ ) δ7.10-8.00 (m, 11H), 4.80-5.20 (m, 2H), 3.60-4.70 (m, 4H), 2.50-3.40 (m, 7H), 2.35 (s, 3H ), 1.40-1.80 (m, 3H), 0.40-1.00 (m, 6H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.22 min., 20.87 min.at 220 nm.

Example 23

N- (4-methylbenzoyl) -N- (1-naphthalenemethyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S)- Methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.00-8.00 (m, 11H), 4.80-5.30 (m, 2H), 3.60-4.70 (m, 2H), 2.40-3.60 (m, 9H), 2.30 (s, 3H ), 1.40-1.80 (m, 3H), 0.40-1.00 (m, 6H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 20.50 min., 21.98 min.at 220 nm.

Example 24

N- (phenylacetyl) -N- (4-methoxybenzyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-methylamino] -3 (S) -methyl Pentylamine

¹ H NMR (CDCl ₃ ) δ6.8-7.4 (m, 9H), 3.6-4.8 (m, 9H), 2.8-3.3 (m, 7H), 0.7-1.4 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 16.43 min at 220 nm.

Example 25

N- (1-naphthoyl) -N- (thiophen-2-yl-methyl) -2 (S)-[(2 (R) -amino-3-mercaptopropyl) -N-acetylamino] -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.8-8.0 (m, 10H), 4.4-4.8 (m, 2H), 3.1-4.1 (m, 6H), 2.8-3.0 (m, 2H), 2.37 (s, 3H ), 0.8-1.6 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 18.62 min at 220 nm.

Example 26. N- (benzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylamino) -3 (S) -methylpentylamine trifluuroacetic acid salt

[Step 1]

N- (1-naphthalenemethyl) -2 (S)-(t-butoxycarbonylamino) -3 (S) -methylpentylamine

The title compound was reacted in the same manner as step 1 of Example 1 using 1-aminomethylnaphthalene (3.00 g, 19.1 mmol) instead of thifen-2-yl-methylamine to give the title compound a yellow oily phase (1.86 g, 28%). Got into:

TLC, Rf = 0.20 (EtOAc / n-Hex = 1/2)

1 H NMR (CDCl ₃ ) δ 7.40-8.25 (m, 7H), 4.65 (d, 1H), 4.15-4.40 (m, 2H), 3.60-3.80 (m, 1H), 2.70-2.90 (m, 2H) , 1.40-1.70 (m, 13H), 0.80-1.00 (m, 6H)

[Step 2]

n- (benzoyl) -N- (1-naphthalenemethyl) -2 (S)-(t-butoxycarbonylamino) -3 (S) -methylpentylamine

The compound prepared in step 1 (0.15 g, 0.423 mmol) was dissolved in 10 mL of anhydrous dimethyl formumamide, benzoyl acid (0.518 g, 0.423 mmol), hydroxybenzotriazole (0.086 g, 0.653 mmol), and diisopropyl carboy Mid (0.081 g, 0.423 mmol) was added. The reaction mixture was stirred overnight at room temperature, then water (100 mL) was added and extracted with ethyl acetate (100 mL × 3). The extracted organic layer was washed with water (100 mL × 2) and brine (100 mL), and excess water was removed with MgSO ₄ and concentrated. The concentrated mixture was column chromatographed with EtOAc / n-hexane (1/3) solution and then concentrated to give the desired product as a clear oily phase (0.09 g, 46%):

TLC Rf = 0.36 (EtOAc / n-Hex = 1/3);

¹ H NMR (CDCl ₃ ) δ 7.2-8.2 (m, 12H), 4.98 (s, 2H), 4.78 (d, 1H), 3.9-4.3 (m, 2H), 2.86 (d, 1H), 1.55 ( s, 9H), 0.65-1.2 (m, 9H):

[Step 3]

N- (benzoyl) -N- (1-naphthalenemethyl) -2 (S)-[N-t-butoxycarbonyl-S-triphenylmethyl-L-cysteinyl] amino-3 (S) -methylpentylamine

The compound prepared in step 2 (0.09 g, 0.196 mmol) was dissolved in 10 mL of 50% trifluoroacetyl acid solution and stirred at room temperature for 2 hours. After the reaction mixture was concentrated, N-methylmorpholine (1 mL) and Nt-butoxycarbonyl-S-triphenylmethyl-L-cysteine (365 mg, 0.784 mmol) in anhydrous dimethylformamide / methylene chloride solvent, Hydroxybenzotriazole (119 mg, 0.882 mmol) and diisopropylcarbodiimide (150 mg, 0.784 mmol) were added and stirred overnight. Water was added to the reaction solution, which was then extracted with ethyl acetate (100 mL × 3). The extracted organic layer was washed with water (100 mL × 2) and brine (100 mL), and excess water was removed with MgSO ₄ and concentrated. The concentrated mixture was column chromatographed with EtOAc / n-Hex (1/3) solution and concentrated to give the desired product in a clear oily phase (121 mg, 76.6%).

TLC Rf = 0.645 (EtOAc / n-Hex = 1/1)

¹ H NMR (CDCl ₃ ) δ 7.0-7.9 (m, 27H), 6.65 (d, 1H), 4.99 (s, 2H), 4.78 (d, 1H), 4.0-4.3 (m, 2H), 3.05 ( d, 1H), 2.6-2.8 (m, 2H), 1.40 (s, 9H), 0.8-1.1 (m, 9H)

[Step 4]

N- (benzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylamino) -3 (S) -methylpentylamine trifluuroacetic acid salt

The compound prepared in step 3 (0.121 g, 0.150 mmol) was dissolved in a solution of trifluoroacetyl acid (9 mL) and triethylsilane (1 mL) and stirred at room temperature for 2 hours. The reaction mixture was concentrated, washed with n-hexane (100 mL × 2) and lyophilized to give the labeling compound as trifluoroacetyl acid salt (52 mg, 61%):

HPLC (gradient, A; 0.1% TFA-Water, B; 0.1% TFA-70% Acetonitrile, 1-21 min (B; 0 → 100%), 21-35 min (100%), 35-55 min (100 → 0%), retention time = 15.49 min at 220 nm;

¹ H NMR (CDCl ₃ ) δ 7.2-8.2 (m, 12H), 4.99 (s, 2H), 3.8-4.4 (m, 3H), 3.05 (bs, 3H), 0.8-1.2 (m, 9H):

[Examples 27-57]

The following analogous peptide derivatives were prepared by reacting the corresponding compounds as starting materials in the same manner as in Example 26.

Example 27

N- (1-naphthoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteininylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.2-8.2 (m, 14H), 4.99 (s, 2H), 3.8-4.4 (m, 3H), 3.05 (bs, 3H), 0.8-1.2 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 20.00 min.

Example 28

N- (3-chlorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ7.2-8.2 (m, 11H), 4.99 (s, 2H), 3.8-4.4 (m, 3H), 3.05 (bs, 3H), 0.8-1.2 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA = Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 19.82 min at 220 nm.

Example 29

N- (3-bromophenylacetyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ 7.0-8.0 (m, 11H), 5.12 (dd, 2H), 4.1-4.3 (m, 3H), 3.64 (dd, 2H), 2.9-3.2 (m, 3H), 0.7-1.4 (m, 9H)

Anaytical HPLC (Gradient, 0.1% TFA in 70% Acetonitrile / 0.1% TFA-Water, 0-1 min (0/100), 1-21 min (0 / 100-100 / 0), 21-35 min (100 / 0), 35-45 min (100 / 0-0 / 100), retention time = 20.26 min at 220 nm.

Example 30

N- (4-methoxybenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteininylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ ) δ6.80-8.20 (m, 11H), 5.10-5.40 (b, 1H), 4.30-4.70 (b, 2H), 3.82 (s, 3H), 2.80-3.30 (m, 2H ), 1.40-2.00 (m, 4H), 0.40-1.40 (m, 8H)

Example 31

N- (Chloacetyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 5.00-5.40 (dd, 2H), 4.00-4.60 (m, 4H), 2.90-3.40 (m, 3H), 1.50-2.00 ( m, 2H), 0.40-1.50 (m, 8H)

Example 32

N- (2,4-difluorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 6.80-8.00 (m, 10H), 5.00 (s, 2H), 4.20-4.60 (m, 2H), 3.80-4.20 (m, 1H), 2.80-3.30 (m, 2H), 1.40-2.00 (m, 2H), 0.40-1.40 (m, 8H)

Example 33

N- (crotonyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 6.20 (d, 1H), 4.80-5.40 (m, 2H), 4.00-4.60 (m, 3H), 2.80-3.30 (m, 3H), 1,72-2.00 (m, 2H), 1.47-1.70 (m, 2H), 0.60-1.47 (m, 8H)

Example 34

N- (cyclopropanenocarbonyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 5.00-5.60 (dd, 2H), 4.10-4.60 (m, 2H), 2.90-3.30 (m, 2H), 1.50-2.00 ( m, 4H), 0.60-1.50 (m, 12H)

Example 35

N- (acetyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 5.10 (dd, 1H), 4.25-4.80 (m, 2H), 4.00-4,23 (m, 1H), 2.90-3.50 ( m, 2H), 2.32 (s, 3H), 1.50-2.00 (m, 3H), 0.60-1.50 (m, 8H)

Example 36

N- (thiophen-2yl-methylcarbonyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 6.80-8.00 (m, 10H), 5.00-5.42 (dd, 2H), 4.22-4.60 (m, 2H), 3.90-4.20 (m, 2H), 2.80-3.20 ( m, 2H), 2.52-2.70 (m, 1H), 1.50-2.00 (m, 2H), 0.40-1.50 (m, 8H)

Example 37

N- (phenoxyacetyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 6.70-8.00 (m, 12H), 5.13 (d, 1H), 4.62-5.08 (dd, 2H), 4.33-4.60 (m, 2H), 4.00-4.30 (m, 1H), 2.90-3.30 (m, 2H), 1.50-2.00 (m, 3H), 0.40-1.50 (m, 8H)

Example 38

N- (thiophen-2yl-carbonyl) -N- (1-naphthalenemethyl) -2 (S) -cysteininylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 6.90-8.00 (m, 10H), 4.20-4.80 (b, 1H), 3.00-3.60 (b, 1H), 1.50-2.00 (m, 2H), 0.40-1.50 ( m, 13H)

Example 39

N- (2-methylbenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.20-8.00 (m, 11H), 5.00 (b, 2H), 4.40 (b, 2H), 2.80-3.30 (m, 2H), 2.40 (s, 3H), 1.50 -2.00 (m, 3H), 0.40-1.50 (m, 8H)

Example 40

N- (pentanoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 5.10 (dd, 2H), 4.41 (m, 2H), 4.00-4.21 (m, 1H), 2.80-3.30 (m, 2H) , 2.40-2.62 (m, 1H), 1.40-2.00 (m, 3H), 1.10-1.40 (m, 4H), 0.60-1.40 (m, 11H)

Example 41

N- (4-methylbenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 11H), 5.10 (m, 2H), 4.20-4.60 (m, 2H), 3.80-4.05 (m, 1H), 2.70-3.20 (m, 2H), 2.32 (s, 3H), 1.40-2.00 (m, 2H), 0.60-1.40 (m, 8H)

Example 42

N- (propionyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.10 (m, 7H), 4.90-5.40 (m, 2H), 4.00-4.90 (m, 2H), 2.80-3.40 (m, 2H), 2.40-2.80 ( m, 2H), 1.50-2.00 (m, 3H), 1.20-1.50 (m, 3H), 0.60-1.50 (m, 8H)

Example 43

N- (2-bromoisobutylyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.20 (m, 7H), 6.00 (d, 1H), 5.20 (m, 1H), 4.20-4.80 (m, 2H), 2.80-3.50 (m, 3H) , 1.90-2.20 (m, 3H), 1.50-1.90 (m, 4H), 0.40-1.50 (m, 9H)

Example 44

N- (2-bromopropionyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 6.80-8.00 (m, 7H), 4.80-5.40 (m, 2H), 3.80-4.80 (m, 2H), 2.80-3.30 (m, 2H), 1.40-2.20 ( m, 4H), 0.40-1.40 (m, 11H)

Example 45

N- (3-bromopropionyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 6.40 (m, 2H), 5.82 (d, 1H), 4.82-5.30 (dd, 2H), 3.90-4.60 (m, 3H) , 2.70-3.20 (m, 3H), 1.40-2.00 (m, 2H), 0.40-1.40 (m, 8H)

Example 46

N- (isobutylcarbonyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 4.90-5.40 (dd, 2H), 4.30-4.60 (m, 2H), 4.00-4.20 (m, 1H), 3.08-3.40 ( m, 1H), 2.70-3.07 (m, 2H), 1.50-2.00 (m, 2H), 1.00-1.50 (m, 6H), 0.60-1.00 (m, 8H)

Example 47

N- (4-chlorobutanecarbonyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 4.90-5.40 (m, 2H), 4.30-4.80 (m, 1H), 3.40-4.00 (m, 2H), 2.40-3.40 ( m, 3H), 2.00-2.20 (m, 1H), 1.50-2.00 (m, 3H), 0.40-1.50 (m, 11H)

Example 48

N- (3-Chloropropionyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 6.42 (bs, 1H), 5.90 (m, 1H), 4.90-5.40 (m, 2H), 4.20-4.60 (m, 2H) , 2.80-3.20 (m, 2H), 1.50-2.00 (m, 2H), 1.20-1.50 (m, 3H), 0.40-1.20 (m, 8H)

Example 49

N- (ethylsuccinyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.10 (m, 7H), 4.97-5.40 (m, 2H), 4.36-4.80 (m, 2H), 3.90-4.35 (m, 3H), 3.72 (dd, 1H), 2.40-3.40 (m, 6H), 1.58 (bs, 1H), 1.20-1.40 (m, 5H), 0.60-1.20 (m, 6H)

Example 50

N- (dichloroacetyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteininylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.05 (m, 7H), 6.20 (s, 1H), 5.00-5.40 (dd, 2H), 4.20-4.60 (m, 2H), 3.12-3.40 (m, 1H), 2.80-3.10 (m, 2H), 1.50-2.00 (m, 2H), 0.60-1.50 (m, 8H)

Example 51

N-cyclobutanecarbonyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.00 (m, 7H), 5.02 (dd, 2H), 4.21-4.70 (m, 2H), 4.10 (bs, 1H), 3.50 (m, 1H), 2.80 -3.37 (m, 2H), 2.10-2.50 (m, 2H), 1.63-2.08 (m, 2H), 1.48-1.63 (m, 1H), 0.60-1.45 (m, 10H)

Example 52

N- (acryloyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.05 (m, 7H), 6.50 (m, 1H), 5.90 (m, 1H), 4.90-5.40 (dd, 2H), 4.15-4.70 (m, 3H) , 2.80-3.40 (m, 3H), 1.50-2.00 (m, 2H), 0.40-1.50 (m, 8H)

Example 53

N- (benzyloxyacetyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.10 (m, 12H), 4.92 (bs, 2H), 4.00-4.90 (m, 5H), 2.80-3.30 (m, 2H), 1.50-2.00 (m, 2H), 0.40-1.50 (m, 10H)

Example 54

N- (4-nitrobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteinylylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.40 (m, 11H), 4.90-5.45 (b, 1H), 4.20-4.90 (m, 2H), 3.00-3.65 (m, 2H), 1.50-2.00 ( m, 2H), 0.40-1.50 (m, 10H)

Example 55

N- (3,4-dichlorobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-8.05 (m, 10H), 5.21 (m, 1H), 4.00-4.80 (b, 1H), 2.80-3.50 (m, 2H), 1.50-2.00 (m, 3H), 0.40-1.50 (m, 10H)

Example 56

N- (3,5-Dinitrobenzoyl) -N- (1-naphthalenemethyl) -2 (S)-(cysteineylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 7.00-9.03 (m, 10H), 5.02 (s, 1H), 4.10-4.70 (m, 2H), 2.90-3.40 (m, 2H), 1.40-2.00 (m, 2H), 0.40-1.40 (m, 10H)

Example 57

N- (3,4,5-trimethoxybenzoyl) -N- (1-naphthalenemethyl) -2 (S) -cysteininylamino) -3 (S) -methylpentylamine

¹ H NMR (CDCl ₃ + TFA) δ 6.80-8.10 (m, 9H), 5.20 (d, 1H), 4.40-4.62 (m, 2H), 4.00-4.38 (m, 1H), 3.90 (s, 3H) , 3.60 (s, 6H), 3.00-3.38 (m, 2H), 1.50-2.00 (m, 2H), 0.40-1.50 (m, 9H)

[Test Example 1]

Proliferation Inhibition Test on Human Cancer Cell Lines

96 wells by floating NCl-H460 (ATCC HTD-177), a human cancer cell line cancerized by K-Ras mutations, in 100 ul medium (Dulbecco's modified Eagle's medium (DMEM) medium containing 10% FBS (fetal bovine serum)) The plate was aliquoted and cultured for 24 hours at 37 ° C. and 5% CO ₂ with only medium added to the control group for absorbance measurement.

After dissolving the compound prepared in Example in a medium to prepare samples of concentrations of 1, 5, 10, 25, 50 uM, these samples were placed in NCl-H460-containing wells and control wells (100 ul) Incubated at 37 ° C., 5% CO _{2 for} 120 hours.

0.1 mg of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-2H-tetrazolium bromide (MTT) was added to all wells in a cultured plate. After incubation for 4 hours at 37 ° C. and 5% CO ₂ , the medium was completely removed and 100 μl of HCl-isopropyl alcohol mixture was added thereto, followed by shaking for 3 minutes. Absorbance was measured at 570 nm with a microplate reader (DL1000, Dynatech Laboratories Co.) and based on this, a concentration (IC ₅₀ ) that inhibits cell growth by 50% was determined. The results are shown in Table 1.

TABLE 1

From the results of Table 1, it can be seen that the compound of formula (I) according to the present invention has a very excellent Ras mutant cell proliferation inhibitory effect.

Claims (4)

Similar peptide derivatives represented by the following general formula (I) or nontoxic salts thereof.

Wherein R ₁ is hydrogen, straight or branched C ₁ -C ₃ alkyl, cyano substituted linear or branched C ₁ -C ₃ alkyl, or acetyl, and A is —CH ₂ — or —CO— . Also, X is a naphthalene, Chi-2-yl, C ₁ - C ₃ alkoxy, phenyl, or C substituted with ₁ - C ₃ alkoxy substituted with C ₁ - C ₃ alkyl and, Y is naphthalene, quinoline, thiophene value -2-yl, thiophen-2-yl-methyl, phenoxymethyl, benzyloxymethyl, ethoxycarbonylethyl, phenyl with or without substituents, phenyl methyl with or without substituents, with or without substituents Is straight or branched C ₁ -C ₅ alkyl or alkenyl, or C ₃ -C ₅ cycloalkyl. The method of claim 1,

Similar peptide derivatives or non-toxic salts thereof, characterized in that selected from the group consisting of tilamine. X-methylamine and a reducing agent are reacted with a compound of formula (II) to prepare a compound of formula (III), and then Y-carboxylic acid is reacted with a fusion agent or Y-carbonyl halide is present. Under reaction to prepare a compound of formula (IV), followed by a deprotection group to prepare a compound of formula (V), followed by an acylation or alkylation reaction to produce a compound of formula (VI), followed by a deprotection group. Method for producing a compound of formula (I) 'characterized in that.

In the above, A, X and Y are as defined in claim 1. In addition, Boc is an amino protecting group, and Pr is a protecting group of a sulfhydryl group. X-methylamine and a reducing agent are reacted with a compound of formula (II) to prepare a compound of formula (III), and then Y-carboxylic acid is reacted with a fusion agent or Y-carbonyl halide is present. To form a compound of formula (IV) by reaction under a deprotection group to prepare a compound of formula (V), followed by an acylation or alkylation to prepare a compound of formula (VI), followed by R1- A method for preparing a compound of formula (I) '', wherein the halide is reacted to produce a compound of formula (VII), followed by a deprotection group.

Wherein A, X and Y are as defined in claim ₁ and R ₁ is straight or branched C ₁ -C ₃ alkyl, cyano substituted straight or branched C ₁ -C ₃ alkyl, or acetyl . In addition, Boc is an amino protecting group, and Pr is a protecting group of a sulfhydryl group.