KR20200081317A - Carboxylic acid-having compounds for PSMA-targeting and use thereof - Google Patents

Abstract

The present invention relates to a ^18F-labeled compound for diagnosing prostate cancer and a use thereof. A carboxylic acid-introduced compound of the present invention forms strong ion-pair bonds (salt-bridge interactions) with arginine residues present at PSMA protein-binding sites, thereby showing high binding affinity, and also having rapid background radiation removal effects in vivo due to the hydrophilic properties of carboxylic acid, and less non-specific binding.

Description

Carboxylic acid-having compounds for PSMA-targeting and use thereof

The present invention relates to ¹⁸ F-labeled compounds for the diagnosis of prostate cancer and uses thereof.

Prostate cancer is the number one mortality rate in men's cancer in the United States, the fifth in Korea, and the second in the world. Prostate cancer usually occurs in men over 50 years old, but has a characteristic that the number of patients increases rapidly with age. It usually progresses slowly, but it is a disease that is extremely difficult to treat when it develops into malignancy and metastasis occurs. Metastasis begins with lymph nodes, pelvic bones, vertebrae and bladder around prostate cancer, and gradually spreads throughout the body.

The methods currently being used for diagnosis include primary prostate-specific antigen test (PSA test) and rectal exam, and medical imaging methods such as rectographic ultrasound, CT, MRI, and whole body bone scan (WBBS). Is being implemented.

However, in most cases, the diagnosis accuracy is low and it is difficult to determine the initial diagnosis of the disease and whether recurrence or metastasis occurs. In addition, there is a disadvantage that it is difficult to distinguish from benign diseases such as prostatic hyperplasia and prostatitis.

Positron emission tomography (PET) is a human imaging method using molecular probes targeting proteins or metabolism specific to a disease, and biochemical changes in the early stages of the disease by using short half-life radioisotopes It has the advantage of observing the disease for early diagnosis, treatment evaluation, and metastasis/recurrence.

Prostate-Specific Membrane Antigen (PSMA) is a cell membrane protein that is specifically overexpressed in prostate cancer cells. It is known to selectively bind well. To date, various radioactive tracers with radioisotope-labeled compounds based on GUL have been studied as prostate cancer specific diagnostic drugs.

Among them, ¹⁸ F-DCFPyL is an ¹⁸ F isotope-labeled GUL compound, and is evaluated as one of the most excellent PET tracers for prostate cancer diagnosis so far reported (Patent Document 1). ¹⁸ F-DCFPyL has a relatively low lipophilicity compared to the previously developed compound ( ¹⁸ F-DCFBzL), and thus has less non-specific binding in vivo and has excellent properties to be removed through the kidneys. ^{The 18} F-DCFPyL or ¹⁸ F-DCFBzL compound has a pyridine and a benzene group, respectively, and these aryl groups have a higher binding capacity than the aryl group-free compound by aryl-cation interaction with the arginine residue at the PSMA protein binding site. It has a characteristic. However, these aryl groups tend to show strong intake in normal tissues such as salivary gland and lacrimal gland by inducing non-specific binding in vivo.

The ¹⁸ F-YC88 compound, which was reported later, is an aryl group-free compound, and the background radioactivity is quickly removed compared to the ¹⁸ F-DCFPyL compound to give a clearer image, while the binding force to the PSMA protein is about 10 times higher than that of the ¹⁸ F-DCFPyL. There is a disadvantage that the prostate cancer intake greatly decreases over time as it decreases (Patent Document 2).

The present inventors confirmed that the compounds in which the carboxylic acid was introduced at the aryl group position of the existing compound not only have excellent binding strength with the PSMA protein, but also show excellent pharmacokinetic properties in vivo due to the hydrophilicity of the carboxylic acid. Did.

US registered patent US 8,778,305 B2 International Patent Publication WO 2017/027870 A1

An object in one aspect of the present invention is to provide a novel compound having excellent binding strength with PSMA protein, and showing excellent pharmacokinetic properties in vivo.

An object in another aspect of the present invention is to provide a composition for diagnosing prostate cancer containing the compound as an active ingredient.

Another object of one aspect of the present invention is to provide a pharmaceutical composition for preventing or treating prostate cancer containing the compound as an active ingredient.

Another object of one aspect of the present invention is to provide a method for diagnosing prostate cancer by administering an effective amount of the compound to a subject.

An object in another aspect of the present invention is to provide a method of treating prostate cancer by administering an effective amount of the compound to a subject.

Another object of the present invention is to provide a use of the compound for the manufacture of a medicament for the treatment of prostate cancer.

In order to achieve the above object,

One aspect of the present invention provides a compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

L ₁ is -(CH ₂ ) _a -, where a is an integer from 2 to 4;

U is a bond, or -C(=O)-;

V is a bond, -NH-, or -C(=O)-;

W is CH, or N;

L ₂ is a bond, or -(CH ₂ ) _b -, where b is an integer from 1 to 3;

X is a bond, -CH ₂ CH ₂ O-, or -C(=O)-;

Y is a bond, N, or CH;

L ₃ is -(CH ₂ ) _c -, where c is an integer from 2 to 4;

L ₄ is -(CH ₂ ) _d -, where d is an integer from 2 to 4;

R is hydrogen, halogen, or CH ₃ ;

Z is -(CH ₂ ) _e -, or -NH-C(=O)-(CH ₂ ) _e -, where e is an integer from 1 to 3;

, 또는

이고;Tz

, or

ego;

L ₅ is -(CH ₂ ) _f -, -(CH ₂ CH ₂ O) _g (CH ₂ ) _h -, or -(CH ₂ O) _i (CH ₂ CH ₂ O) _j (CH ₂ ) _k- , Where f, g, h, i, j and k are independently integers from 1 to 5;

m and n are independently integers of 0 or 1,

는 -H, 또는 부재이고,If m is an integer of 0,

Is -H, or absent,

는 -H, 또는 부재이고; 및If n is an integer of 0,

Is -H, or absent; And

F is ¹⁸ F.

Another aspect of the present invention provides a compound represented by Formula 2, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 2]

In Chemical Formula 2,

L ₁ is -(CH ₂ ) _a -, where a is an integer from 2 to 4;

U is a bond, or -C(=O)-;

V is a bond, -NH-, or -C(=O)-;

W is CH, or N;

L ₂ is a bond, or -(CH ₂ ) _b -, where b is an integer from 1 to 3;

X is a bond, -CH ₂ CH ₂ O-, or -C(=O)-;

Y is a bond, N, or CH;

L ₃ is -(CH ₂ ) _c -, where c is an integer from 2 to 4;

L ₄ is -(CH ₂ ) _d -, where d is an integer from 2 to 4;

R is hydrogen, halogen, or CH ₃ ;

Z is -(CH ₂ ) _e -, or -NH-C(=O)-(CH ₂ ) _e -, where e is an integer from 1 to 3;

T is -C≡CH, or -N ₃ ; And

n is an integer of 0 or 1,

는 -H, 또는 부재이다.If n is an integer of 0,

Is -H, or absent.

Another aspect of the present invention provides a composition for diagnosing prostate cancer containing the compound, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating prostate cancer containing the compound, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect of the present invention provides a method for diagnosing prostate cancer by administering an effective amount of the compound to a subject.

Another aspect of the present invention provides a method of treating prostate cancer by administering an effective amount of the compound to a subject.

Another aspect of the invention provides the use of the compound for the manufacture of a medicament for the treatment of prostate cancer.

The carboxylic acid-introduced compounds of the present invention have strong binding affinity by forming a strong ion-pair bond (Salt Bridge Interaction) with an arginine residue at the PSMA protein binding site, and are fast in vivo due to the hydrophilic nature of the carboxylic acid. It has the effect of removing background radioactivity and low non-specific binding.

Hereinafter, the present invention will be described in detail.

Meanwhile, the embodiments of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below. In addition, embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art. Furthermore, "including" a component throughout the specification means that other components may be further included, rather than excluding other components, unless otherwise specified.

One aspect of the invention,

It provides a compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

L ₁ is -(CH ₂ ) _a -, where a is an integer from 2 to 4;

U is a bond, or -C(=O)-;

V is a bond, -NH-, or -C(=O)-;

W is CH, or N;

L ₂ is a bond, or -(CH ₂ ) _b -, where b is an integer from 1 to 3;

X is a bond, -CH ₂ CH ₂ O-, or -C(=O)-;

Y is a bond, N, or CH;

L ₃ is -(CH ₂ ) _c -, where c is an integer from 2 to 4;

L ₄ is -(CH ₂ ) _d -, where d is an integer from 2 to 4;

R is hydrogen, halogen, or CH ₃ ;

Z is -(CH ₂ ) _e -, or -NH-C(=O)-(CH ₂ ) _e -, where e is an integer from 1 to 3;

, 또는

이고;Tz

, or

ego;

L ₅ is -(CH ₂ ) _f -, -(CH ₂ CH ₂ O) _g (CH ₂ ) _h -, or -(CH ₂ O) _i (CH ₂ CH ₂ O) _j (CH ₂ ) _k- , Where f, g, h, i, j and k are independently integers from 1 to 5;

m and n are independently integers of 0 or 1,

는 -H, 또는 부재이고,If m is an integer of 0,

Is -H, or absent,

는 -H, 또는 부재이고; 및If n is an integer of 0,

Is -H, or absent; And

F is ¹⁸ F.

When R is halogen, the halogen may be a radioactive isotope.

The compound represented by Chemical Formula 1 may be a compound represented by Chemical Formula 1-1.

[Formula 1-1]

In Chemical Formula 1-1,

L ₁ , U, V, W, L ₂ , X, Z, Tz, L ₅ and F are as defined above; And

Y is a bond, NH, or CH ₂ .

Another aspect of the invention,

It provides a compound represented by Formula 2, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 2]

In Chemical Formula 2,

L ₁ is -(CH ₂ ) _a -, where a is an integer from 2 to 4;

U is a bond, or -C(=O)-;

V is a bond, -NH-, or -C(=O)-;

W is CH, or N;

L ₂ is a bond, or -(CH ₂ ) _b -, where b is an integer from 1 to 3;

X is a bond, -CH ₂ CH ₂ O-, or -C(=O)-;

Y is a bond, N, or CH;

L ₃ is -(CH ₂ ) _c -, where c is an integer from 2 to 4;

L ₄ is -(CH ₂ ) _d -, where d is an integer from 2 to 4;

R is hydrogen, halogen, or CH ₃ ;

Z is -(CH ₂ ) _e -, or -NH-C(=O)-(CH ₂ ) _e -, where e is an integer from 1 to 3;

T is -C≡CH, or -N ₃ ; And

n is an integer of 0 or 1,

는 -H, 또는 부재이다.If n is an integer of 0,

Is -H, or absent.

The compound represented by Formula 1,

It may be any one compound selected from the group consisting of the following formula 1e, 1f, 1g, 1h, 1i, 1j, 1k and 1l.

,

,

,

,

,

,

,

,

,

,

,

,

및

.

And

.

The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., 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, trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. Get from The types of pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and eye Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube Late, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, Glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate produced by dissolving a derivative of Formula I in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic acid or inorganic acid It can be prepared by filtration and drying, or by distilling the solvent and excess acid under reduced pressure and drying to crystallize under an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the inexpensive compound salt, and evaporating and drying the filtrate. At this time, it is suitable to manufacture sodium, potassium or calcium salts as metal salts. Further, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).

Furthermore, the present invention includes all of the compounds represented by Formula 1 and pharmaceutically acceptable salts thereof, as well as solvates, optical isomers, and hydrates that can be prepared therefrom.

Another aspect of the present invention provides a composition for diagnosing prostate cancer containing a compound, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound diagnoses prostate cancer by selectively binding to Prostate-Specific Membrane Antigen (PSMA), which is overexpressed in prostate cancer cells.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating prostate cancer containing the compound, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The pharmaceutical composition using the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient may be administered parenterally, and parenteral administration may be administered by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. How to do.

At this time, in order to formulate a formulation for parenteral administration, a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed with water with a stabilizer or a buffer to prepare a solution or suspension, and it is administered in ampoules or vials. Can be produced. The composition may be sterile and/or contain preservatives, stabilizers, hydrating or emulsifying accelerators, adjuvants such as salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, conventional methods of mixing, granulation It can be formulated according to the chemical or coating method.

Another aspect of the present invention provides a method for diagnosing prostate cancer by administering an effective amount of the compound to a subject.

Another aspect of the present invention provides a method of treating prostate cancer by administering an effective amount of the compound to a subject.

Another aspect of the invention provides the use of the compound for the manufacture of a medicament for the treatment of prostate cancer.

The carboxylic acid-introduced compounds of the present invention have strong binding affinity by forming a strong ion pair bond (Salt Bridge Interaction) with an arginine residue at the PSMA protein binding site, and are fast in vivo due to the hydrophilic properties of carboxylic acid. It is characterized by a background radioactive removal effect and a low non-specific binding, which is directly supported by the examples and experimental examples described below.

Hereinafter, the present invention will be described in detail through examples and experimental examples described below.

However, the examples and experimental examples to be described later are merely examples of the present invention, and the present invention is not limited thereto.

< Example 1> Preparation of compounds 3b and 3c

Preparation of compound 3b

Compound 3a (5.2 g, 10.7 mmol) was dichloromethane (100 mL), cooled to 0° C., then tert-butyl bromoacetate (1.9 mL, 12.8 mmol) was added slowly. Triethylamine (2.2 mL, 16.0 mmol) was slowly added while maintaining 0° C., and the temperature was gradually increased to room temperature and stirred. After stirring for 3 hours, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (50 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 3b (3.36 g, 52%).

MS (ESI) m/z 602 [M+H] ⁺

Preparation of compound 3c

Compound 3a (150 mg, 0.31 mmol) was dissolved in ethanol (2.0 mL) and tert-butyl acrylate (0.05 mL, 0.034 mmol) was added slowly. After stirring for 12 hours, it was concentrated under reduced pressure and the concentrate was separated by column chromatography (4% methanol/dichloromethane) to obtain compound 3c (73 mg, 39%).

MS (ESI) m/z 616 [M+H] ⁺

< Example 2> Preparation of compounds 1a, 1b, 1c

Preparation of compound 1a

Step 1:

Compound 3a (300 mg, 0.62 mmol) was dissolved in dichloromethane (6.0 mL), cooled to 0° C., and then acetyl chloride (0.05 mL, 0.68 mmol) was slowly added. Triethylamine (0.26 mL, 1.85 mmol) was slowly added while maintaining 0° C., and the temperature was gradually increased to room temperature and stirred. After stirring for 1 hour, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (30 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 4a (265 mg, 81%).

MS (ESI) m/z 552 [M+Na] ⁺

Step 2:

Compound 4a (100 mg, 0.19 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane solution (1.5 mL) and stirred at room temperature for 3 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1a (60 mg, 88%).

MS (ESI) m/z 362 [M+H] ⁺

Preparation of compound 1b

Step 1:

Compound 3b (100 mg, 0.17 mmol) obtained in Example 1 was dissolved in dichloromethane (30 mL), cooled to 0° C., and then acetyl chloride (0.02 mL, 0.26 mmol) was slowly added. Triethylamine (0.07 mL, 0.51 mmol) was slowly added while maintaining 0° C., and the temperature was gradually increased to room temperature and stirred. After stirring for 1 hour, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (30 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (4% methanol/dichloromethane) to obtain compound 4b (79 mg, 74%).

MS (ESI) m/z 666 [M+Na] ⁺

Step 2:

Compound 4b (59 mg, 0.09 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane solution (1.0 mL) and stirred at room temperature for 3 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1b (20 mg, 52%).

MS (ESI) m/z 442 [M+Na] ⁺

Preparation of compound 1c

Step 1:

Compound 3c (60 mg, 0.10 mmol) obtained in Example 1 was dissolved in dichloromethane (1.2 mL), cooled to 0° C., and then acetyl chloride (0.01 mL, 0.15 mmol) was slowly added. Triethylamine (0.04 mL, 0.30 mmol) was slowly added while maintaining 0° C., and the temperature was gradually increased to room temperature and stirred. After stirring for 1 hour, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (30 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (4% methanol/dichloromethane) to obtain compound 4c (63 mg, 98%).

MS (ESI) m/z 680 [M+Na] ⁺

Step 2:

Compound 4c (50 mg, 0.08 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane solution (1.0 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1c (24 mg, 73%).

MS (ESI) m/z 434 [M+H] ⁺

< Example 3> Preparation of compounds 1d, 1e

Preparation of compound 1d

Step 1:

Dissolve 6-(N-Fmoc-amino)hexanoic acid (72 mg, 0.25 mmol) in dimethylformamide (1.0 mL), add HOBT (42 mg, 0.31 mmol) and TBTU (99 mg, 0.31 mmol) Stirred at room temperature. Diisopropylethylamine (0.11 mL, 0.62 mmol) was added to the reaction mixture, and 3a (100 mg, 0.21 mmol) diluted in dimethylformamide (1.0 mL) was slowly added. After stirring for 1 hour, an aqueous ammonium chloride solution (50 mL) was added, and then an organic compound was extracted with ethyl acetate (30 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 5a (147 mg, 87%).

MS (ESI) m/z 845 [M+Na] ⁺

Step 2:

Compound 5a (353 mg, 0.43 mmol) obtained in step 1 was dissolved in a 20% piperidine/dichloromethane solution (5.0 mL) and stirred at room temperature for 2 hours. Water (50 mL) was added, and then the organic compound was extracted with dichloromethane (30 mL, 2 times). The collected organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (NH silica gel, FUJI SILYSIA CHEMICAL LTD) (1% methanol/dichloromethane) to obtain compound 6a (260 mg, 99%). .

MS (ESI) m/z 601 [M+H] ⁺

Step 3:

Dissolve 4-(p-tolyl)butanoic acid (11 mg, 0.06 mmol) in dichloromethane (5.0 mL), add HOBT (11 mg, 0.08 mmol) and TBTU (26 mg, 0.08 mmol) and stir at room temperature. Did. Diisopropylethylamine (0.03 mL, 0.15 mmol) was added to the reaction mixture, and 6a (30 mg, 0.05 mmol) diluted in dichloromethane (2.0 mL) was slowly added. After stirring for 1 hour, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (20 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 7a (32 mg, 84%).

MS (ESI) m/z 783 [M+Na] ⁺

Step 4:

Compound 7a (32 mg, 0.042 mmol) obtained in step 3 was dissolved in 70% TFA/dichloromethane solution (0.5 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1d (5.4 mg, 22%).

MS (ESI) m/z 593 [M+H] ⁺

Preparation of compound 1e

Step 1:

Dissolve 6-(N-Fmoc-amino)hexanoic acid (212 mg, 0.60 mmol) in dichloromethane (5.0 mL), cool to 0 °C, then N,N'-dicyclohexylcarbodiimide (DCC, 113 mg, 0.55 mmol) was added, and then 3b (300 mg, 0.50 mmol) was added and stirred at room temperature for 1 hour. The reaction solution was filtered under reduced pressure, concentrated, and the concentrate was separated by column chromatography (2% methanol/dichloromethane) to obtain compound 5b (261 mg, 56%).

MS (ESI) m/z 959 [M+Na] ⁺

Step 2:

Compound 5b (123 mg, 0.13 mmol) obtained in step 1 was dissolved in a 20% piperidine/dichloromethane solution (2.0 mL) and stirred at room temperature for 2 hours. Water (50 mL) was added, and then the organic compound was extracted with dichloromethane (20 mL, 2 times). The collected organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (NH silica gel, FUJI SILYSIA CHEMICAL LTD) (1% methanol/dichloromethane) to obtain compound 6b (88 mg, 94%). .

MS (ESI) m/z 715 [M+H] ⁺

Step 3:

Dissolve 4-(p-tolyl)butanoic acid (4 mg, 0.02 mmol) in dichloromethane (5.0 mL), add HOBT (4 mg, 0.03 mmol) and TBTU (10 mg, 0.03 mmol) and stir at room temperature. Did. Diisopropylethylamine (0.01 mL, 0.06 mmol) was added to the reaction mixture, and 6b (15 mg, 0.02 mmol) diluted in dichloromethane (2.0 mL) was slowly added. After stirring for 1 hour, water (30 mL) was added, and then an organic compound was extracted with dichloromethane (20 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 7b (15 mg, 82%).

MS (ESI) m/z 897 [M+Na] ⁺

Step 4:

Compound 7b (18 mg, 0.02 mmol) obtained in step 3 was dissolved in 60% TFA/dichloromethane solution (0.5 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1e (5.6 mg, 42%).

MS (ESI) m/z 651 [M+H] ⁺

< Example 4> Preparation of compound 1f

Step 1:

4-Pentinoic acid (82 mg, 0.83 mmol) with dichloromethane (10 mL), cooled to 0° C., and then added N,N′-dicyclohexylcarbodiimide (DCC, 190 mg, 0.91 mmol), followed by compound 3b obtained in Example 1 (500 mg, 0.83 mmol) ) Was added and stirred at room temperature for 1 hour. The reaction solution was filtered under reduced pressure, concentrated, and the concentrate was separated by column chromatography (30% ethyl acetate/ n -hexane) to obtain compound 8 (290 mg, 52%).

MS (ESI) m/z 682 [M+H] ⁺

Step 2:

Compound 8 (100 mg, 0.15 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane solution (2 mL) and stirred at room temperature for 3 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 2a (37 mg, 55%).

MS (ESI) m/z 458 [M+H] ⁺ , 456 [MH] ^-

Step 3:

After dissolving compound 2a (13 mg, 0.028 mmol) and F-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ (5 mg, 0.028 mmol) in step 2 in ethanol (1.0 mL), 1 M CuSO ₄ aqueous solution (6 μL, 0.006 mmol) and 2 M sodium ascorbate aqueous solution (Na-Asc. 5 μL, 0.009 mmol) were sequentially added and stirred for 1 hour. The reaction solution was filtered, concentrated under reduced pressure, and then separated using high performance liquid chromatography (HPLC). The purified solution was lyophilized to obtain compound 1f (8 mg, 44%) as a white solid.

¹ H NMR (400 MHz, D ₂ O) δ 1.20-1.38 (m, 2H), 1.39-1.56 (m, 2H), 1.58-1.71 (m, 1H), 1.72-1.84 (m, 1H), 1.86- 1.95 (m, 1H), 2.06-2.16 (m, 1H), 2.45 (t, J = 7.2 Hz, 2H), 2.68 (t, J = 6.8 Hz, 1H), 2.84 (t, J = 6.4 Hz, 1H ), 3.00 (q, J = 6.8 Hz, 2H), 3.28-3.36 (m, 2H), 3.59-3.62 (m, 5H), 3.68-3.70 (m, 1H), 3.91 (t, J = 4.4 Hz, 2H), 4.01 (s, 1H), 4.09-4.21 (m, 3H), 4.45 (t, J = 4.0 Hz, 1H), 4.56-4.60 (m, 3H), 7.89 (s, 1H)

MS (ESI) m/z 635 [M+H] ⁺ , 633 [MH] ^-

< Example 5> Preparation of compound 1g

Step 1:

Azidoacetic acid (18 mg, 0.18 mmol) is dichloromethane (5.0 mL) and cooled to 0 °C. N,N'-dicyclohexylcarbodiimide (DCC, 41 mg, 0.20 mmol) was added and compound 3b (109 mg, 0.18 mmol) obtained in Example 1 was added thereto, followed by stirring at room temperature for 1 hour. After filtering the reaction solution, the solvent was removed under reduced pressure, and then column chromatography (40% ethyl acetate/ n -hexane) was performed to obtain compound 9 (80 mg, 64%).

Step 2:

Compound 9 (80 mg, 0.11 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane solution (1.0 mL) and stirred for 4 hours. After adding the solution to the ethyl ether to catch the precipitate, ethyl ether (20 mL) was further added and then separated using a centrifuge. The precipitate was dissolved in water, separated using high performance liquid chromatography (HPLC), and then lyophilized to obtain compound 2b (30 mg, 56%).

Step 3:

Compound 2b (8 mg, 0.017 mmol) and 3-(2-(2-fluoroethoxy)ethoxy)prop-1-ene (3 mg, 0.017 mmol) obtained in step 2 above were used in ethanol (1 mL). After dissolving in, 1 M CuSO ₄ aqueous solution (4 μL, 0.003 mmol) and 2 M sodium ascorbate aqueous solution (Na-Asc. 3 μL, 0.005 mmol) were added and stirred for 1 hour. After filtering the reaction solution, the solvent was removed under reduced pressure, separated using high performance liquid chromatography (HPLC), and then lyophilized to obtain compound 1g (10 mg, 95%).

< Example 6> Preparation of compounds 10a and 10b

Preparation of 10a:

Propargyl amine (160 mg, 2.87 mmol) was dissolved in dichloromethane (10 mL) and brought to 0° C., then triethylamine (0.6 mL, 4.3 mmol) was added slowly. Tert-Butyl bromoacetate (0.62 g, 3.16 mmol) was dissolved in dichloromethane (10 mL), slowly added, and stirred at room temperature for 18 hours. The reaction was terminated by adding water (20 mL) and the organic compound was extracted with dichloromethane (20 mL, 2 times). After dehydration with anhydrous sodium sulfate, it was concentrated under reduced pressure, and the concentrate was subjected to column chromatography (2% methanol/dichloromethane) to obtain compound 10a (0.46 g, 55%).

Preparation of 10b:

Dissolve 2-(2-azidoethoxy)ethylamine (100 mg, 0.77 mmol) in dichloromethane (30 mL), cool to 0 °C, and add tert-butyl bromoacetate (0.14 mL, 0.92 mmol) gave. While maintaining 0°C, triethylamine (0.32 mL, 2.31 mmol) was slowly added and the reaction solution temperature was gradually raised to room temperature and stirred. After stirring for 1 hour, the reaction solution was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 10b (78 mg, 42%).

< Example 7> Preparation of compounds 10c and 10d

Preparation of 10c

After dissolving proparzyl amine (500 mg, 9.07 mmol) in ethanol (20 mL) and cooling to 0° C., tert-butyl acrylate (1.16 g, 9.07 mmol) was slowly added and stirred at room temperature for 18 hours. The solvent was removed under reduced pressure, and the concentrate was subjected to column chromatography (2% methanol/dichloromethane) to obtain compound 10c (1.4 g, 84%).

Manufacturing of 10d

2-(2-azidoethoxy)ethylamine (1.0 g, 7.68 mmol) was dissolved in ethanol (200 mL) and tert-butyl acrylate (1.23 mL, 8.45 mmol) was added slowly. After stirring for 12 hours, it was concentrated under reduced pressure and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 10d (1.07 g, 54%).

< Example 8> Preparation of compound 11

H-Glu(OtBu)-OtBu.HCl (500 mg, 1.69 mmol) was dissolved in dichloromethane (25 mL) and cooled to -10 °C. Diisopropylethylamine (0.7 mL, 4.05 mmol) was added and 4-nitrophenyl chloroformate (340 mg, 1.69 mmol) was added slowly. After stirring at -10°C for 10 minutes, it was further stirred at room temperature for 20 minutes. After confirming the reaction by thin plate chromatography (TLC), cool again to -10 °C, add triethylamine (0.56 mL, 4.05 mmol), add H-Glu(OH)-OtBu.HCl (330 mg, 1.60 mmol), It was stirred at room temperature for 1 hour. Water (25 mL) was added to the reaction solution, and the organic compound was extracted with dichloromethane (25 mL, 2 times). After removing water with anhydrous sodium sulfate, it was concentrated under reduced pressure and the concentrate was subjected to column chromatography (5% methanol/dichloromethane). Separated with to obtain compound 11 (600 mg, 72%).

MS (ESI) m/z 489 [M+H] ⁺ , 487 [MH] ^-

< Example 9> Preparation of compounds 1h and 1i

Preparation of compound 1h

Step 1:

Compound 11 (300 mg, 0.61 mmol) obtained in Example 8 was dissolved in dichloromethane (30 mL), cooled to 0° C., and then N,N'-dicyclohexylcarbodiimide (DCC, 139 mg, 0.68 mmol) ) Was added, and then compound 10b (225 mg, 0.92 mmol) obtained in Example 6 was added and stirred at room temperature for 1 hour. The reaction solution was filtered under reduced pressure, concentrated, and the concentrate was separated by column chromatography (2% methanol/dichloromethane) to obtain compound 12a (413 mg, 94%).

MS (ESI) m/z 751 [M+Na] ⁺

Step 2:

Compound 12a (300 mg, 0.42 mmol) obtained in step 1 was dissolved in methanol (20 mL), palladium (10% Palladium on carbon, 22 mg) was added, and the mixture was stirred at room temperature for 1 hour under hydrogen. The reaction solution was filtered under reduced pressure, concentrated, and the concentrate was separated by column chromatography (NH silica gel, FUJI SILYSIA CHEMICAL LTD) (1% methanol/dichloromethane) to obtain compound 13a (231 mg, 80%).

MS (ESI) m/z 689 [M+H] ⁺

Step 3:

Dissolve 4-(p-tolyl)butanoic acid (16 mg, 0.09 mmol) in dichloromethane (5 mL), add HOBT (15 mg, 0.11 mmol) and TBTU (35 mg, 0.11 mmol) and stir at room temperature. Did. Diisopropylethylamine (0.04 mL, 0.22 mmol) was added to the reaction mixture, and then 13a (50 mg, 0.08 mmol) obtained in step 2 diluted in dichloromethane (5 mL) was slowly added. After stirring for 1 hour, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (30 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 14a (51 mg, 83%).

MS (ESI) m/z 871 [M+Na] ⁺

Step 4:

Compound 14a (50 mg, 0.06 mmol) obtained in step 3 was dissolved in 70% TFA/dichloromethane solution (1.0 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1h (18 mg, 49%).

MS (ESI) m/z 625 [M+H] ⁺

Preparation of compound 1i

Step 1:

Compound 11 (300 mg, 0.61 mmol) obtained in Example 8 was dissolved in dichloromethane (30 mL), cooled to 0° C., and then N,N'-dicyclohexylcarbodiimide (DCC, 139 mg, 0.68 mmol) ) Was added, and then 10d (238 mg, 0.92 mmol) obtained in Example 7 was added and stirred at room temperature for 1 hour. The reaction solution was filtered under reduced pressure, concentrated, and the concentrate was separated by column chromatography (2% methanol/dichloromethane) to obtain compound 12b (327 g, 73%).

MS (ESI) m/z 751 [M+Na] ⁺

Step 2:

Compound 12b (300 mg, 0.41 mmol) obtained in step 1 was dissolved in methanol (20 mL), palladium (10% Palladium on carbon, 22 mg) was added, and the mixture was stirred at room temperature for 1 hour under hydrogen. The reaction solution was filtered under reduced pressure, concentrated, and the concentrate was separated by column chromatography (NH silica gel, FUJI SILYSIA CHEMICAL LTD) (1% methanol/dichloromethane) to obtain compound 13b (197 mg, 68%).

MS (ESI) m/z 703 [M+H] ⁺

Step 3:

Dissolve 4-(p-tolyl)butanoic acid (30 mg, 0.17 mmol) in dichloromethane (10 mL), add HOBT (29 mg, 0.21 mmol) and TBTU (68 mg, 0.21 mmol) and stir at room temperature. Did. Diisopropylethylamine (0.07 mL, 0.43 mmol) was added to the reaction mixture, and 13b (100 mg, 0.14 mmol) obtained in Step 2, diluted in dichloromethane (5 mL), was slowly added. After stirring for 1 hour, water (50 mL) was added, and then the organic compound was extracted with dichloromethane (30 mL, 2 times). The combined organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 14b (120 mg, 98%).

MS (ESI) m/z 885 [M+Na] ⁺

Step 4:

Compound 14b (56 mg, 0.06 mmol) obtained in step 3 was dissolved in 70% TFA/dichloromethane solution (1.0 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 1i (28 mg, 68%).

MS (ESI) m/z 639 [M+H] ⁺

< Example 10> Preparation of compound 1j, 1k

Preparation of compound 1j

Step 1:

Compound 11 (500 mg, 1.02 mmol) obtained in Example 8 was dichloromethane After dissolving in (10 mL), the mixture was cooled to 0° C., and then N,N'-dicyclohexylcarbodiimide (DCC, 230 mg, 1.12 mmol) was added, and compound 10a (0.17 g, 1.02 mmol) obtained in Example 6 was added. ) Was added and stirred at room temperature for 1 hour. The reaction solution was filtered, concentrated under reduced pressure, and the concentrate was separated by column chromatography (30% ethyl acetate/ n -hexane) to obtain compound 15a (450 mg, 69%).

MS (ESI) m/z 640 [M+H] ⁺

Step 2:

Compound 15a (75 mg, 0.12 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane solution (1.0 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized separated solution was obtained as a white solid, compound 2c (14 mg, 29%).

MS (ESI) m/z 416 [M+H] ⁺ , 414 [MH] ^-

Step 3:

After dissolving compound 2c (14 mg, 0.034 mmol) and F-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ (6 mg, 0.034 mmol) in step 2 in ethanol (1 mL), 1 M CuSO ₄ aqueous solution (7 μL, 0.007 mmol) and 2 M sodium ascorbate aqueous solution (Na-Asc. 5 μL, 0.01 mmol) were sequentially added and stirred for 1 hour. The reaction solution was filtered, concentrated under reduced pressure, and then separated using high performance liquid chromatography (HPLC). The purified solution was lyophilized to obtain compound 1j (13 mg, 65%) as a white solid.

¹ H NMR (400 MHz, D ₂ O) δ 1.88-2.01 (m, 2H), 2.09-2.19 (m, 2H), 2.43-2.49 (m, 3H), 2.67-2.73 (m, 1H), 3.61- 3.64 (m, 5H), 3.71 (t, J = 4.0 Hz, 1H), 3.90-3.93 (m, 2H), 4.12 (s, 1H), 4.19-4.24 (m, 2H), 4.27 (d, J = 2.8 Hz, 1H), 4.45-4.48 (m, 1H), 4.56-4.60 (m, 3H), 4.65 (d, J = 6.0 Hz, 1H), 4.74 (s, 1H), 7.94 (s, 0.4H) , 8.02 (s, 0.6H)

MS (ESI) m/z 593 [M+H] ⁺ , 591 [MH] ^-

Preparation of compound 1k

Step 1:

Compound 11 (83 mg, 0.45 mmol) obtained in Example 8 was dichloromethane After dissolving in (15 mL), cooled to 0 ^o C, then N,N'-dicyclohexylcarbodiimide (DCC, 102 mg, 0.50 mmol) was added and compound 10c (220 mg, 0.45) obtained in Example 7 was added. mmol) and stirred at room temperature for 1 hour. The reaction solution was filtered and the solvent was removed under reduced pressure, followed by column chromatography (30% ethyl acetate/ n -hexane) to obtain compound 15c (190 mg, 64%).

Step 2:

Compound 15c (170 mg, 0.26 mmol) obtained in step 1 was dissolved in 70% TFA/dichloromethane (2.0 mL) and stirred for 4 hours. After adding the reaction solution to the ethyl ether to catch the precipitate, ethyl ether (20 mL) was further added, and then the precipitate was separated using a centrifuge. The precipitate was dissolved in water, separated using high performance liquid chromatography (HPLC), and lyophilized to obtain compound 2d (83 mg, 74%).

Step 3:

After dissolving compound 2d (20 mg, 0.046 mmol) and F-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ (8 mg, 0.046 mmol) obtained in step 2 in ethanol (2 mL), 1 M CuSO ₄ aqueous solution (9 μL, 0.009 mmol) and 2 M sodium ascorbate aqueous solution (Na-Asc. 7 μL, 0.014 mmol) were added and stirred for 1 hour. After filtering the reaction solution, the solvent was removed under reduced pressure, and then separated using high performance liquid chromatography (HPLC) and lyophilized to obtain compound 1k (20 mg, 70%).

<Example 11> Preparation of compound 1l

Step 1:

Compound 3b (500 mg, 0.83 mmol) obtained in Example 1 and Fmoc-Lys(Z)-OH (340 mg, 0.83 mmol) were dissolved in dichloromethane (10 mL), cooled to 0° C., and then N,N'. -Dicyclohexylcarbodiimide (DCC, 190 mg, 0.913 mmol) was added and stirred for 1 hour. After filtering the reaction solution, the solvent was removed under reduced pressure, and then subjected to column chromatography (50% ethyl acetate/n-hexane) to obtain compound 16 (500 mg, 55%).

MS (ESI) m/z 1087 [M+H] ⁺

Step 2:

Compound 16 (490 mg, 0.45 mmol) obtained in Step 1 was dissolved in a solution of 20% piperidine/dichloromethane (10 mL) and stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, and then separated by column chromatography (NH silica gel) to obtain compound 17 (240 mg, 61%).

MS (ESI) m/z 865 [M+H] ⁺

Step 3:

Dissolve pentinoic acid (12 mg, 0.121 mmol) in dichloromethane (2.0 mL) and add HOBt (16 mg, 0.121 mmol), TBTU (39 mg, 0.121 mmol), diisopropylethylamine (28 μL, 0.162 mmol). It was put in order and stirred for 10 minutes. Compound 17 (70 mg, 0.081 mmol) obtained in Step 2 above diluted with dichloromethane (1.0 mL) was added, stirred at room temperature for 1 hour, and then water was added. The organic compound was extracted using dichloromethane, treated with anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (3% methanol/dichloromethane) to obtain compound 18 (70 mg, 91%).

MS (ESI) m/z 945 [M+H] ⁺ , 967 [M+Na] ⁺

Step 4:

After dissolving compound 18 (80 mg, 0.0847 mmol) and F-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ (15 mg, 0.0847 mmol) in step ₃ in ethanol (1 mL), 1 M CuSO ₄ aqueous solution (17 μL, 0.017 mmol) and 2 M sodium ascorbate aqueous solution (13 μL, 0.026 mmol) were sequentially added and stirred for 1 hour. The reaction solution was filtered, concentrated under reduced pressure, and then separated by column chromatography (2% methanol/dichloromethane) to obtain compound 19 (76 mg, 80%).

MS (ESI) m/z 1122 [M+H] ⁺

Step 5:

Compound 19 (60 mg, 0.053 mmol) obtained in step 4 was dissolved in methanol (5 mL), palladium (10% Palladium on carbon, 22 mg) was added, and the mixture was stirred at room temperature for 2 hours under hydrogen. The reaction solution was filtered under reduced pressure and concentrated to remove the solvent to obtain compound 20 (35 mg, 67%).

MS (ESI) m/z 988 [M+H] ⁺

Step 6:

Dissolve 4-(p-tolyl)butanoic acid (8 mg, 0.0425 mmol) in dichloromethane (5 mL), HOBt (8 mg, 0.0531 mmol), TBTU (17 mg, 0.0531 mmol), diisopropylethylamine ( 13 μL, 0.0708 mmol) was added in sequence and stirred for 10 minutes. Compound 20 (35 mg, 0.0354 mmol) obtained in step 5 was dissolved in dichloromethane (2 mL) and stirred at room temperature overnight. The solvent was removed under reduced pressure and separated by column chromatography (2% methanol/dichloromethane) to obtain compound 21 (40 mg, 98%).

MS (ESI) m/z 1148 [M+H] ⁺

Step 7:

Compound 21 (40 mg, 0.0348 mmol) obtained in Step 6 was dissolved in 70% trifluoroacetic acid/dichloromethane solution (1.0 mL) and stirred at room temperature for 4 hours. The reaction solution was slowly dropped into ethyl ether to form a precipitate, and ethyl ether (20 mL) was further added. The resulting precipitate was separated using a centrifuge, purified by high performance liquid chromatography (HPLC), and then the lyophilized solution was lyophilized to obtain 1 l of a white solid (24 mg, 74%).

MS (ESI) m/z 924 [M+H] ⁺ , 922 [MH] ⁺

< Example 12> Compound [ ¹⁸ F] Preparation of 1f

Chromafix® (HCO ₃ ^-) to give a flow of distilled water (3 mL) was passed through a ^[18 F] fluoride solution (120 mCi) was then flushed with ethanol (1 mL). Krytofix222-potassium methanesulfonate (10 mg) was dissolved in ethanol (1 mL) to pass Chromafix® and the solution was blown with nitrogen gas at 100° C. to remove the solvent. Dissolve MsO-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ (1 mg) in t-butanol (0.5 mL) and place it in a reaction vessel with [ ¹⁸ F]fluoride, then react at 100°C for 10 minutes. Was prepared (preparation of ¹⁸ F-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ ). After the reaction, nitrogen gas was gently blown at 100° C., the solvent was removed, and then ethanol (0.3 mL) was added to dissolve. To this solution, add compound 2a (3 mg) dissolved in distilled water (0.1 mL), add 0.5 M CuSO ₄ aqueous solution (5 μL) and 0.5 M sodium ascorbate aqueous solution (10 μL) in sequence, and react for 10 minutes at room temperature. Ordered. Distilled water (2 mL) was added to the reaction mixture, the solution was filtered, and then separated by HPLC to obtain compound [ ¹⁸ F] 1f (34 mCi).

< Example 13> Compound [ ¹⁸ F] Preparation of 1j

Chromafix® (HCO ₃ ^-) to give a flow of distilled water (3 mL) was passed through a ^[18 F] fluoride solution (80 mCi) was then flushed with ethanol (1 mL). Krytofix222-potassium methanesulfonate (10 mg) was dissolved in ethanol (1 mL) to pass Chromafix® and the solution was blown with nitrogen gas at 100° C. to remove the solvent. Dissolve MsO-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ (1 mg) in t-butanol (0.5 mL) and place it in a reaction vessel with [ ¹⁸ F]fluoride, then react at 100°C for 10 minutes. Was prepared (preparation of ¹⁸ F-CH ₂ CH ₂ (OCH ₂ CH ₂ ) ₂ -N ₃ ). After the reaction, nitrogen gas was blown at 100°C to remove the solvent, followed by dissolving in ethanol (0.3 mL). To this solution, add compound 2c (3 mg) dissolved in distilled water (0.1 mL), add 0.5 M CuSO ₄ aqueous solution (5 μL) and 0.5 M sodium ascorbate aqueous solution (10 μL) in sequence, and react for 10 minutes at room temperature. Ordered. Distilled water (2 mL) was added to the reaction mixture, the solution was filtered, and then separated by HPLC to obtain compound [ ¹⁸ F] 1j (12 mCi).

< Comparative example 1> DCFPyL Produce

Step 1:

Dissolve 6-fluoronicotinic acid (100 mg, 0.71 mmol) and 2,3,5,6-tetrafluorophenol (130 mg, 0.78 mmol) in dioxane (2.0 mL) and N,N-dicyclohex Silcarbodiimide (DCC, 146 mg, 0.71 mmol) was added to the reaction solution and stirred at room temperature for 2 hours. The precipitate formed after the reaction was filtered and washed with dichloromethane. The filtrate was concentrated under reduced pressure, followed by column chromatography (5% ethyl acetate/n-hexane) to obtain compound 22 (136 mg, 47%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.09-7.16 (m, 2H), 8.58 (ddd, J = 2.5 Hz, 7.3 Hz, 8.7 Hz, 1H), 9.10 (d, J = 2.4 Hz, 1H);

MS (ESI) m/z 290 [M+H] ⁺

Step 2:

Compound 22 (100 mg, 0.35 mmol) and compound 3a (202 mg, 0.42 mmol) obtained in step 1 above were dichloromethane. (5.0 mL) and triethylamine (0.146 mL, 1.1 mmol) was added to the reaction solution and stirred at room temperature for 20 minutes. After the reaction, distilled water was added, and then an organic compound was extracted using dichloromethane. The combined organic solution was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and obtained column 23 (138 mg, 66%) as a colorless oil by column chromatography (2% methanol/dichloromethane).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.37 (s, 9H), 1.42 (s, 9H), 1.43 (s, 9H), 1.48-1.62 (m, 3H), 1.65-1.70 (m, 1H), 1.72-1.87 (m, 3H), 1.98-2.07 (m, 1H), 2.25-2.38 (m, 2H), 3.32-3.39 (m, 1H), 3.54-3.62 (m, 1H), 4.13-4.21 (m , 2H), 5.46 (d, J = 7.6 Hz, 1H), 5.88 (d, J = 8.0 Hz, 1H), 7.00 (dd, J = 3.0 Hz, 8.4 Hz, 1H), 7.93 (s, 1H), 8.42 (m, 1H), 8.84 (d, J =2.4 Hz, 1H);

MS (ESI) m/z 611 [M+H] ⁺ , 633 [M+Na] ⁺

Step 3:

Compound 23 (100 mg, 0.16 mmol) obtained in Step 2 was dissolved in 60% TFA/dichloromethane solution (1.0 mL) and stirred at room temperature for 4 hours. After adding the reaction solution to the ethyl ether to catch the precipitate, ethyl ether (20 mL) was further added, and then the precipitate was separated using a centrifuge. The precipitate was dissolved in water, separated using high performance liquid chromatography (HPLC), and then lyophilized to obtain the compound DCFPyL (56 mg, 79%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.41-1.47 (m, 2H), 1.58-1.65 (m, 2H), 1.67-1.75 (m, 1H) 1.79-1.95 (m, 2H), 2.06-2.15 ( m, 1H), 2.06-2.15 (m, 1H), 2.44 (t, J = 7.2 Hz, 2H), 3.37 (t, J = 6.6 Hz, 2H),

4.13-4.18 (m, 2H), 7.17 (d, J = 8.4 Hz, 1H), 8.25 (tt, J = 1.2 Hz, 8.0 Hz, 1H), 8.51 (d, J = 1.6 Hz, 1H);

MS (ESI) m/z 465 [M+Na] ⁺

< Comparative example 2> [ ¹²⁵ I] Synthesis of 20 compounds

Step 1:

Triphosgene (21 mg, 0.071 mmol) was dissolved in dichloromethane (5 mL), 4-iodineaniline (45 mg, 0.21 mmol) dissolved in dichloromethane (5 mL) was slowly added at 0°C, and then triethylamine (0.57) mL, 0.41 mmol) was added, followed by stirring at 0°C for 30 minutes. Compound 3a (100 mg, 0.21 mmol) dissolved in dichloromethane (10 mL) was slowly added at 0°C, triethylamine (0.57 mL, 0.41 mmol) was added, and the temperature was slowly raised to room temperature and stirred for 5 hours. The mixture was concentrated under reduced pressure, and column chromatography (2% methanol/dichloromethane) gave compound 24 (66 mg, 44%) as a colorless oil.

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.20-1.27 (m, 2H), 1.37 (s, 9H), 1.40 (s, 9H), 1.44 (s, 9H), 1.47-1.57 (m, 2H), 1.71-1.81 (m, 2H), 1.83-1.91 (m, 1H), 2.03-2.11 (m, 1H), 2.37 (sext, J = 8.2 Hz, 2H), 3.01-3.07 (m, 1H), 3.51- 3.56 (m, 1H), 3.97-4.01 (m, 1H), 4.26-4.32 (m, 1H), 5.75 (d, J = 7.2 Hz, 1H), 6.31 (q, J = 3.4 Hz, 1H), 6.40 (d, J = 8.0 Hz, 1H), 7.27 (d, J = 8.8 Hz, 2H), 7.52 (d, J = 8.8 Hz, 2H), 7.90 (s, 1H);

¹³ C NMR (100 MHz, CDCl ₃ ) δ 24.5, 27.1, 27.8, 27.9, 28.0, 29.6, 31.7, 32.0, 39.1, 53.8, 54.9, 81.0, 81.8, 83.6, 83.7, 120.2, 137.5, 140.2, 155.6, 158.5 , 171.8, 172.0, 175.3; MS (ESI) m/z 733 [M+H] ⁺

Step 2:

After dissolving compound 24 (50 mg, 0.068 mmol) synthesized in step 1 in 1,4-dioxane (1.0 mL), hexamethylditin ((Me ₃ Sn) ₂ , 0.043 mL, 0.21 mmol) and bis( Triphenylphosphine)palladium dichloride (4.8 mg, 5 mol%) was added one after another and then stirred at 110° C. for 90 minutes. The mixture was cooled to room temperature, added with an aqueous potassium fluoride solution (50 mL), stirred for 1 hour, and then filtered. The filtrate was extracted with ethyl acetate and the organic solution was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and used column chromatography (triethylamine:ethyl acetate:n-hexane, 1:40:59). This gave compound 25 (28 mg, 53%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 0.25 (s, 9H), 1.22-1.29 (m, 2H), 1.38 (s, 9H), 1.41 (s, 9H), 1.43 (s, 9H), 1.48- 1.59 (m, 2H), 1.72-1.78 (m, 1H), 1.81-1.91 (m, 1H), 2.05-2.13 (m, 2H), 2.34-2.43 (m, 2H), 3.04-3.09 (m, 1H) ), 3.51-3.55 (m, 1H), 4.04 (pent, J = 4.9 Hz, 1H), 4.33 (sext, J = 4.5 Hz, 1H), 5.73 (d, J = 6.8 Hz 1H), 6.23 (br s , 1H), 6.32 (d, J = 8.4 Hz, 1H), 7.35 (d, J = 8.0 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 7.73 (s, 1H);

¹³ C NMR (100 MHz, CDCl ₃ ) δ -9.5, 24.2, 27.4, 27.8, 27.9, 28.0, 29.7, 31.8, 32.1, 39.1, 53.7, 54.7, 80.9, 81.7, 83.5, 118.4, 133.6, 136.2, 140.4, 155.9, 158.3, 171.9, 172.2, 175.1; MS (ESI) m/z 771 [M+2H] ⁺

Step 3:

After dissolving compound 25 (0.1 mg) obtained in step 2 in ethanol (0.250 mL), add sodium [ ¹²⁵ I] iodide aqueous solution (3.2 mCi, 0.050 mL) and stir at room temperature. After adding 1 N aqueous HCl solution (0.10 mL), 3% H ₂ O ₂ was added and stirred at room temperature for 10 minutes. An aqueous sodium thiosulfate solution (0.1 M, 0.20 mL) was added to the reaction mixture, distilled water (18 mL) was added, and the solution was passed through C-18 Sep-Pak, followed by distilled water (20 mL). After flowing acetonitrile (2.0 mL) to C-18 Sep-Pak, nitrogen was blown into the solution to remove acetonitrile.

Step 4:

Dichloromethane (0.2 mL) and trifluoroacetic acid (0.8 mL) were sequentially added to the reaction mixture obtained in step 3 and stirred at room temperature for 20 minutes. The reaction solvent was removed by blowing nitrogen, and then distilled water (2.0 mL) was added, and the solution was separated by high performance liquid chromatography (HPLC) to obtain compound [ ¹²⁵ I] 26 (1.1 mCi, 24%).

HPLC conditions:

Column, XTerra MS C18 (250 mm×10 mm); Mobile phase, 30% acetonitrile/water (0.1% TFA); Flow rate, 5 mL/min; UV, 254 mm; Retention time, 10.4 min.

< Reference example 1> Prostate cancer cell line and nude mouse preparation

The human prostate cancer cell line (22RV1) was purchased from the American Type Culture Collection (ATCC) and used. People prostate cancer cell line, PC3 PIP (PSMA ⁺⁾ and PC3 flu (PSMA ^-) cell lines Dr. Used by Martin G. Pomper (Johns Hopkins Medical School, Baltimore, MD). The human prostate cancer cell line was used as a culture medium containing 10% fetal bovine serum (FBS) and 1% antibiotic/antifungal agent in RPMI1640 medium. PC3 PIP (PSMA ⁺⁾ and PC3 flu (PSMA ^-) cell lines when cultured, the cells were cultured by the addition of puromycin (Puromycin) of the further 2 μg / mL concentration.

As a test animal, male nude mice 6 weeks old (Narabio, Seoul, Korea) were used.

< Experimental Example 1> Binding ability Measure

In order to confirm the binding capacity of the Example compound of the present invention to the PSMA protein, experiments were conducted as follows.

As a buffer solution, 1% concentration of BSA (bovine serum albumin) was added to RPMI1640 medium.

In a container containing 22RV1 cells (5×10 ⁴ ), put [ ¹²⁵ I] 26 (0.1 nM) obtained in Comparative Example 2, and formula 1 at 9 concentrations (1.00 x 10 ^-4 to 1.00 x 10 ^-12 M) Each of the compounds was added and stirred at 37°C for 2 hours. After stirring was completed, each was washed three times with PBS solution (2 mL), and then the radiation dose was measured with a gamma counter (2480 WIZARD2 Gamma Counter PerkinElmer Co., MA). The 50% inhibitory concentration (IC ₅₀ ) for each compound was calculated using the GraphPad Prism (GraphPad Software, Inc., CA) program.

Table 1 below is a table showing the binding affinity (IC ₅₀ ) of each compound, and in Table 1 below, DCFPyL is a compound obtained in Comparative Example 1. (Chen, Y. et al., Clin Cancer Res. 2011, 17(24), 7645-7653)

compound IC ₅₀ (Mean±SD, nM) DCFPyL (Comparative Example 1) 30.71±10.18 1a 136.76±8.47 1b 12.16±0.93 1c 12.46±3.51 1e 13.86±0.55 1f 12.21±0.46 1 g 12.12±0.17 1h 108.21±1.87 1j 78.88±5.36 1k 197.84±28.32 1l 86.06±3.13

As shown in Table 1 above,

It can be seen that the compounds 1b and 1c of Example 2 of the present invention are compounds having a carboxylic acid, and have about 11 times higher binding strength to the PSMA protein than compound 1a without a carboxylic acid. It is one of three arginine residues (R463) called arginine patch in the binding region of the PSMA protein and strong carboxylic acid bond of the compound of formula 1 of the present invention (Salt Bridge Interaction) It can be interpreted as forming.

It is known that the DCFPyL compound of Comparative Example 1 has a pyridine bond, and it is aryl-cation-interacting with the arginine residue (R463) of the PSMA protein to increase the binding force.

However, the compounds of Example 2 of the present invention having a carboxyl group have a binding strength of about 2.5 times higher than that of DCFPyL of Comparative Example 1, which is more ionic bonding between arginine cation and carboxylate anion than aryl-positron interaction. Strongness is consistent with previous research results.

In addition, it can be seen that Compounds 1e , 1f , and 1g also show a similar level of binding ability to PSMA protein as Compounds 1b and 1c .

That is, the carboxylic acid of the compound of Formula 1 of the present invention not only greatly improves the binding strength with the PSMA protein, but also increases the polarity of the compound, thereby reducing the non-specific binding in vivo and removing it more rapidly.

As described above, the present invention has been described in detail through preferred examples and experimental examples, but the scope of the present invention is not limited to specific examples, and should be interpreted by the appended claims. In addition, those skilled in the art should understand that many modifications and variations are possible without departing from the scope of the present invention.

Claims (8)

A compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Chemical Formula 1,
L ₁ is -(CH ₂ ) _a -, where a is an integer from 2 to 4;
U is a bond, or -C(=O)-;
V is a bond, -NH-, or -C(=O)-;
W is CH, or N;
L ₂ is a bond, or -(CH ₂ ) _b -, where b is an integer from 1 to 3;
X is a bond, -CH ₂ CH ₂ O-, or -C(=O)-;
Y is a bond, N, or CH;
L ₃ is -(CH ₂ ) _c -, where c is an integer from 2 to 4;
L ₄ is -(CH ₂ ) _d -, where d is an integer from 2 to 4;
R is hydrogen, halogen, or CH ₃ ;
Z is -(CH ₂ ) _e -, or -NH-C(=O)-(CH ₂ ) _e -, where e is an integer from 1 to 3;
Tz

, or

ego;
L ₅ is -(CH ₂ ) _f -, -(CH ₂ CH ₂ O) _g (CH ₂ ) _h -, or -(CH ₂ O) _i (CH ₂ CH ₂ O) _j (CH ₂ ) _k- , Where f, g, h, i, j and k are independently integers from 1 to 5;

m and n are independently integers of 0 or 1,
If m is an integer of 0,

Is -H, or absent,
If n is an integer of 0,

Is -H, or absent; And

F is ¹⁸ F.
According to claim 1,
When R is halogen, the halogen is a radioactive isotope compound, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.
According to claim 1,
The compound represented by Formula 1 is a compound, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, characterized in that it is a compound represented by Formula 1-1:
[Formula 1-1]

In Chemical Formula 1-1,
L ₁ , U, V, W, L ₂ , X, Z, Tz, L ₅ and F are as defined in claim 1; And
Y is a bond, NH, or CH ₂ .
A compound represented by Formula 2 below, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof:
[Formula 2]

In Chemical Formula 2,
L ₁ is -(CH ₂ ) _a -, where a is an integer from 2 to 4;
U is a bond, or -C(=O)-;
V is a bond, -NH-, or -C(=O)-;
W is CH, or N;
L ₂ is a bond, or -(CH ₂ ) _b -, where b is an integer from 1 to 3;
X is a bond, -CH ₂ CH ₂ O-, or -C(=O)-;
Y is a bond, N, or CH;
L ₃ is -(CH ₂ ) _c -, where c is an integer from 2 to 4;
L ₄ is -(CH ₂ ) _d -, where d is an integer from 2 to 4;
R is hydrogen, halogen, or CH ₃ ;
Z is -(CH ₂ ) _e -, or -NH-C(=O)-(CH ₂ ) _e -, where e is an integer from 1 to 3;
T is -C≡CH, or -N ₃ ; And

n is an integer of 0 or 1,
If n is an integer of 0,

Is -H, or absent.
According to claim 1,
The compound represented by Formula 1 is a compound characterized in that it is any one compound selected from the group consisting of the following Formulas 1e, 1f, 1g, 1h, 1i, 1j, 1k and 1l, stereoisomers thereof, and hydrates thereof , Or a pharmaceutically acceptable salt thereof:

,

,

,

,

,

,

And

.
A composition for diagnosing prostate cancer comprising the compound of claim 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 6,
The compound is a composition for diagnosing prostate cancer, characterized in that it selectively binds to a prostate-specific membrane membrane antibody (Prostate-Specific Membrane Antigen, PSMA) that overexpresses prostate cancer cells.
A pharmaceutical composition for preventing or treating prostate cancer comprising the compound of claim 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.